Degenerative cervical myelopathy is a common cause of spinal cord injury,with longer symptom duration and higher myelopathy severity indicating a worse prognosis.While numerous studies have investigated serological bi...Degenerative cervical myelopathy is a common cause of spinal cord injury,with longer symptom duration and higher myelopathy severity indicating a worse prognosis.While numerous studies have investigated serological biomarkers for acute spinal cord injury,few studies have explored such biomarkers for diagnosing degenerative cervical myelopathy.This study involved 30 patients with degenerative cervical myelopathy(51.3±7.3 years old,12 women and 18 men),seven healthy controls(25.7±1.7 years old,one woman and six men),and nine patients with cervical spondylotic radiculopathy(51.9±8.6 years old,three women and six men).Analysis of blood samples from the three groups showed clear differences in transcriptomic characteristics.Enrichment analysis identified 128 differentially expressed genes that were enriched in patients with neurological disabilities.Using least absolute shrinkage and selection operator analysis,we constructed a five-gene model(TBCD,TPM2,PNKD,EIF4G2,and AP5Z1)to diagnose degenerative cervical myelopathy with an accuracy of 93.5%.One-gene models(TCAP and SDHA)identified mild and severe degenerative cervical myelopathy with accuracies of 83.3%and 76.7%,respectively.Signatures of two immune cell types(memory B cells and memory-activated CD4^(+)T cells)predicted levels of lesions in degenerative cervical myelopathy with 80%accuracy.Our results suggest that peripheral blood RNA biomarkers could be used to predict lesion severity in degenerative cervical myelopathy.展开更多
Parkinson’s disease(PD)is a common neurodegenerative disorder with profound impact on patients’quality of life and long-term health,and early detection and intervention are particularly critical.In recent years,the ...Parkinson’s disease(PD)is a common neurodegenerative disorder with profound impact on patients’quality of life and long-term health,and early detection and intervention are particularly critical.In recent years,the search for precise and reliable biomarkers has become one of the key strategies to effectively address the clinical challenges of PD.In this paper,we systematically evaluated potential biomarkers,including proteins,metabolites,epigenetic markers,and exosomes,in the peripheral blood of PD patients.Protein markers are one of the main directions of biomarker research in PD.In particular,α‑synuclein and its phosphorylated form play a key role in the pathological process of PD.It has been shown that aggregation ofα-synuclein may be associated with pathologic protein deposition in PD and may be a potential marker for early diagnosis of PD.In terms of metabolites,uric acid,as a metabolite,plays an important role in oxidative stress and neuroprotection in PD.It has been found that changes in uric acid levels may be associated with the onset and progression of PD,showing its potential as an early diagnostic marker.Epigenetic markers,such as DNA methylation modifications and miRNAs,have also attracted much attention in Parkinson’s disease research.Changes in these markers may affect the expression of PD-related genes and have an important impact on the onset and progression of the disease,providing new research perspectives for the early diagnosis of PD.In addition,exosomes,as a potential biomarker carrier for PD,are able to carry a variety of biomolecules involved in intercellular communication and pathological regulation.Studies have shown that exosomes may play an important role in the pathogenesis of PD,and their detection in blood may provide a new breakthrough for early diagnosis.It has been shown that exosomes may play an important role in the pathogenesis of PD,and their detection in blood may provide new breakthroughs in early diagnosis.In summary,through in-depth evaluation of biomarkers in the peripheral blood of PD patients,this paper demonstrates the important potential of these markers in the early diagnosis of PD and in the study of pathological mechanisms.Future studies will continue to explore the clinical application value of these biomarkers to promote the early detection of PD and individualized treatment strategies.展开更多
BACKGROUND Patients with acute-on-chronic liver failure(ACLF)experience severe immune dysfunction.Liver transplantation(LT)significantly improves survival outcomes.However,the characteristics of peripheral blood lymph...BACKGROUND Patients with acute-on-chronic liver failure(ACLF)experience severe immune dysfunction.Liver transplantation(LT)significantly improves survival outcomes.However,the characteristics of peripheral blood lymphocyte subsets(PBLSs)in this patient population are not well defined,and the dynamics of immune reconstitution post-LT are insufficiently understood.AIM To characterize PBLSs in patients with ACLF prior to LT and to evaluate PBLS reconstitution after LT.METHODS Clinical data from patients undergoing LT in the Transplantation Center,The Third Xiangya Hospital from January 2022 to December 2023 were analyzed retrospectively.Our cohort comprised 44 patients with ACLF,16 patients with acute decompensation of cirrhosis,and 23 patients with compensated cirrhosis.Twenty healthy volunteers were included as controls.PBLSs were evaluated across all groups.The relationship between PBLSs and post-LT prognosis was assessed,and dynamic changes in PBLSs among patients with ACLF were analyzed at different time points.RESULTS Patients with ACLF exhibited a marked reduction in PBLSs compared with healthy volunteers.Natural killer(NK)cell counts were further reduced in patients with ACLF when compared with patients with compensated cirrhosis.PBLSs did not correlate with the etiology or severity of ACLF or with established liver failure scores.Following LT,a rapid restoration of NK cells and B cells was observed in patients with ACLF.However,the cluster of differentiation(CD)3+T cell and CD4+T cell counts decreased 14 days post-LT and subsequently returned to preoperative levels by day 21.CONCLUSION Patients with ACLF exhibited markedly reduced PBLSs,with decreased NK cells potentially linked to progression from compensated cirrhosis to liver failure.NK and B cell were rapidly restored after LT.展开更多
In the context of global aging,mild behavioral impairment(MBI)is present in 48.9%of patients with mild cognitive impairment(MCI).MBI,a neurobehavioral syndrome in the elderly,is an independent risk factor for cognitiv...In the context of global aging,mild behavioral impairment(MBI)is present in 48.9%of patients with mild cognitive impairment(MCI).MBI,a neurobehavioral syndrome in the elderly,is an independent risk factor for cognitive decline and is closely related to peripheral blood biomarkers associated with Alzheimer's disease,offering new diagnostic and interventional avenues for early MCI.To summarize evidence on peripheral blood biomarkers related to MBI and their underlying mechanisms involving neuroinflammation,tau pathology,and oxidative stress,a systematic review of studies published between 2015 and 2024 was conducted.MBI is closely associated with peripheral blood biomarker changes.Neuroinflammatory markers like glial fibrillary acidic protein and neurofilament light indicate astrocyte activation and neural circuit disruption,with glial fibrillary acidic protein levels correlating with impulse dyscontrol scores.Chitinase-3-like protein 1,a marker of blood-brain barrier integrity,exacerbates neuroinflammation and is linked to depressive symptoms and hippocampal atrophy.Elevated phosphorylated tau proteins in blood correlate with brain tau deposition,increasing the risk of MBI and impairing cognition.Oxidative stress markers damage neurons and disrupt neurotransmission,and concurrent alterations in malondialdehyde and superoxide dismutase levels significantly elevate the risk of MBI.The correlation between MBI and biomarkers offers new diagnostic and interventional directions for early MCI.Future research should standardize MBI assessment,conduct longitudinal studies,explore biomarker-MBI relationships,investigate psychosocial impacts,and develop advanced detection methods.展开更多
Objectives To compare respiratory parameters of peripheral blood mononuclear cell mitochondria and iron metabolism indicators in patients with different NYHA functional classes of ischemic heart failure(HF).Methods Th...Objectives To compare respiratory parameters of peripheral blood mononuclear cell mitochondria and iron metabolism indicators in patients with different NYHA functional classes of ischemic heart failure(HF).Methods This single center, prospective, non-blinded study enrolled 20 patients with diagnosed chronic HF of ischemic genesis with reduced and mildly reduced left ventricle ejection fraction. The maximum oxygen consumption at the peak of the exercise test(VO2peak), iron metabolism parameters and respiratory activity of peripheral blood mononuclear cell mitochondria were assessed.Results Among the patients, a half of individuals were diagnosed with iron deficiency. Subgroups of patients with different HF severity did not significant differ in VO2peak(P=0.209), serum iron(P=0.468) and ferritin(P=0.235) levels. But there was a trend in increasing in these parameters with increasing NYHA HF functional class. Respiratory control coefficient(RC) in NADdependent and FAD-dependent mitochondrial oxidation were lower in patients with NYHA HF Ⅲ functional class compared to individuals with NYHA HF I functional class(P=0.028 and P=0.040, respectively). Serum iron(P=0.026), ferritin(P=0.045)levels, transferrin saturation(P=0.006) were negatively correlated with RC in NAD-dependent mitochondrial oxidation.Conclusions In aggravation of ischemic HF NYHA FC, there is a decrease in RC of PBMC mitochondria during the oxidation of NAD-dependent and FAD-dependent substrates. In the whole sample, patients with laboratory-confirmed iron deficiency accounted a half of the total number. Iron metabolism parameters had a paradoxical inverse relationship with the level of RC in PBMC mitochondria of patients with HF.展开更多
Objective:To explore the accuracy and stability of the results of peripheral blood routine tests at different time points after anticoagulation and standing,providing a scientific basis for actual clinical work.Method...Objective:To explore the accuracy and stability of the results of peripheral blood routine tests at different time points after anticoagulation and standing,providing a scientific basis for actual clinical work.Methods:In this study,30 patients who visited the hospital in October 2023 were randomly selected and divided into two groups(15 cases in each group).The same collection method was used for routine blood tests.The tests were performed after anticoagulation and standing for 5 minutes,1 hour,and 5 minutes,2 hours respectively,and the routine blood test indicators at different time points were compared.Results:After comparison,there were no significant differences in the results of routine blood tests at 5 minutes after mixing,anticoagulating,and standing peripheral blood and those at 1 hour and 2 hours(P>0.05).Conclusion:The results of peripheral blood after mixing,anticoagulating,and standing for 5 minutes are stable compared with those after standing for 1 hour and 2 hours.In actual work,the pre-test turnaround time can be appropriately extended.展开更多
AIM:To investigate the possible relationship between inflammatory biomarkers in the peripheral blood of patients with branch retinal vein occlusion(BRVO).METHODS:A total of 63 BRVO patients were enrolled in this cross...AIM:To investigate the possible relationship between inflammatory biomarkers in the peripheral blood of patients with branch retinal vein occlusion(BRVO).METHODS:A total of 63 BRVO patients were enrolled in this cross-sectional observational study.Meanwhile,63 age-and gender-matched cataract patients were included as controls.Complete blood count and biochemical tests were performed,and inflammatory biomarkers including platelet to lymphocyte ratio(PLR),red blood cell distribution width to albumin ratio(RAR),neutrophil to lymphocyte ratio(NLR),systemic immune inflammation index(SII),and monocyte to high density lipoprotein cholesterol ratio(MHR)were compared between the two groups.RESULTS:There were no significant differences between the two groups in terms of age,sex,and prevalence of diabetes mellitus.Compared with the controls,patients with BRVO had a higher prevalence of hypertension and higher body mass index(BMI).Red blood cell distribution width(RDW),triglycerides,MHR,NLR,and RAR were elevated,whereas lymphocyte count and high-density lipoprotein were decreased in the BRVO group.Multivariate logistic regression analysis revealed that NLR(adjusted OR=1.686,95%CI 1.075-2.646),RAR(adjusted OR=8.930,95%CI 1.911-41.730),and body mass index(BMI;adjusted OR=1.174,95%CI 1.010-1.365)were significantly associated with the risk of BRVO.In the receiver operating characteristic analysis,the area under the curve for NLR,RAR,and BMI were 0.602,0.630,and 0.603,respectively.The sensitivity and specificity were 61.9%and 60.3%,38.1%and 82.5%,and 61.9%and 57.1%,respectively.CONCLUSION:Peripheral blood inflammatory biomarkers are elevated in BRVO patients,suggesting systemic inflammation involvement.NLR,RAR,and BMI are positively correlated with BRVO.Monitoring NLR and RAR and strict weight control may be beneficial for the prevention and treatment of BRVO.展开更多
Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnosti...Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnostic value for hepatitis B virus-associated hepatocellular carcinoma(HBV-HCC). Methods: We enrolled 197 patients, 111 with HBV-HCC, 51 with chronic hepatitis B(CHB), and 35 healthy controls(HCs). The methylation status of GLDC promoter in peripheral mononuclear cells(PBMCs) was identified by methylation specific polymerase chain reaction(MSP). The mRNA expression was examined using real-time quantitative polymerase chain reaction(q PCR). Results: The methylation frequency of the GLDC promoter was significantly lower in HBV-HCC patients(27.0%) compared to that in CHB patients(68.6%) and HCs(74.3%)( P < 0.001). The methylated group had lower alanine aminotransferase level( P = 0.035) and lower rates of tumor node metastasis(TNM) Ⅲ/Ⅳ( P = 0.043) and T3/T4( P = 0.026). TNM stage was identified to be an independent factor for GLDC promoter methylation. GLDC mRNA levels in CHB patients and HCs were significantly lower than those in HBV-HCC patients( P = 0.022 and P < 0.001, respectively). GLDC mRNA levels were significantly higher in HBV-HCC patients with unmethylated GLDC promoters than those with methylated GLDC promoters( P = 0.003). The diagnostic accuracy of alpha-fetoprotein(AFP) combined with GLDC promoter methylation for HBV-HCC was improved compared with that of AFP alone(AUC: 0.782 vs. 0.630, P < 0.001). In addition, GLDC promoter methylation was an independent predictor for overall survival of HBV-HCC patients( P = 0.038). Conclusions: The methylation frequency of GLDC promoter was lower in PBMCs from HBV-HCC patients than that from patients with CHB and HCs. The combination of AFP and GLDC promoter hypomethylation significantly improved the diagnostic accuracy of HBV-HCC.展开更多
As an efficient immunostimulant,chitosan oligosaccharide(COS)can enhance the immunity of teleosts;however,the underlying molecular mechanisms still require elucidation.Competing endogenous RNAs(ceRNAs)are vital regula...As an efficient immunostimulant,chitosan oligosaccharide(COS)can enhance the immunity of teleosts;however,the underlying molecular mechanisms still require elucidation.Competing endogenous RNAs(ceRNAs)are vital regulators in the immune response,but their roles in half-smooth tongue sole(Cynoglossus semilaevis)remain unclear.In this study,for the first time,we studied whole-transcriptome expression profiles and analyzed ceRNA networks in peripheral blood leukocytes of half-smooth tongue sole treated with COS.A total of 19 circRNAs(DE-circRNAs),18 miRNAs(DE-miRNAs)and 50 previously identified lncRNAs(DElncRNAs)were differentially expressed after COS stimulation.The DE-lncRNAs and DE-miRNAs targeted numerous immunity-related genes,and were enriched in important pathways,including MAPK and Toll-like receptor signaling pathways,suggesting the immunoregulatory roles of COS.Furthermore,we constructed circRNA-miRNA-mRNA and lncRNA-miRNA-mRNA regulatory networks using DE-circRNAs,DE-miRNAs,DE-lncRNAs,and DE-mRNAs.Additionally,a ceRNA network with immunity-related DEmRNAs was constructed,showing that 3 DE-circRNAs,12 DE-lncRNAs,and 29 DEGs exhibited crosstalk through 9 DE-miRNAs.Intriguingly,a DE-miRNA in the ceRNA network,miR-144-3p,was targeted by DE-lncRNA tnrc6a,and negatively regulated the genes of inhibitor of nuclear factor kappa B kinases(IKKs)(ikbkg,ikbkb,and ikbip)and c3ar1.Ikbkg,ikbkb,and c3ar1 were significantly up-regulated in macrophages stimulated by LPS.It could be inferred that ncRNAs participated in the immune and inflammatory response by acting as ceRNAs after COS stimulation in teleosts.These findings indicate that COS could enhance the immunity of teleosts by regulating ncRNAs,and lay the foundation for further practical application of COS in aquaculture.展开更多
BACKGROUND Gastric cancer is one of the most common malignant tumors worldwide,and surgical resection is one of the main ways to treat gastric cancer.However,the immune status of postoperative patients is crucial for ...BACKGROUND Gastric cancer is one of the most common malignant tumors worldwide,and surgical resection is one of the main ways to treat gastric cancer.However,the immune status of postoperative patients is crucial for prognosis and survival,and immune cells play an important role in this process.Therefore,it is helpful to understand the immune status of postoperative patients by evaluating the levels of peripheral blood immune cells,especially total T cells(CD3+),helper T cells(CD3+CD4+),and suppressor T cells(CD3+CD8+),and its relationship to sur-vival.AIM To analyzed the immune cells in peripheral blood of patients with gastric cancer after surgery,detect the levels of total T cells,helper T cells and suppressor T cells.METHODS A total of 58 patients with gastric cancer who received surgical treatment were included in the retrospective study.Flow cytometry was used to detect the level of peripheral blood immune cells and analyze the correlation between total T cells,helper T cells and inhibitory T cells.To explore the relationship between these immune markers and patient survival.RESULTS The results showed that the levels of total T cells,helper T cells,and suppressor T cells changed in patients after gastric cancer surgery.There was a significant positive correlation between total T cells,helper T cells and suppressor T cells(r=0.35,P<0.01;r=0.56,P<0.01).However,there was a negative correlation between helper T cells and suppressor T cells(r=-0.63,P<0.01).Follow-up showed that the survival rate of patients in the high-level total T cell group was significantly higher than that in the low-level group(28.87±24.98 months vs 18.42±16.21 months).The survival curve shows that the curve of patients in the high-level group is shifted to the upper right,and that of the low-level group is shifted downward.There was no significant difference between the levels of helper T cells and suppressor T cells and patient survival time.CONCLUSION By detecting peripheral blood immune cells with flow cytometry,we can initially evaluate the immune status of patients after gastric cancer surgery and initially explore its relationship with patient survival.展开更多
Objective： To study the genes expression profile differences in the peripheral blood between esophageal carcinoma patients and normal subjects using the gene chip technique and screen out the esophageal early concera...Objective： To study the genes expression profile differences in the peripheral blood between esophageal carcinoma patients and normal subjects using the gene chip technique and screen out the esophageal early conceration associated genes. Methods： The total RNA was extracted and purified in the peripheral blood obtained from the patients with esophageal carcinoma and normal subjects. The first strand of cDNA was synthesized through retro-transcription and labeled with Cy5 and Cy3 fluorescence as probes. The mixed probes were hybridized with a piece of 4096 double dot human whole gene chip. The acquired image was analyzed by microarrav suite software using a digital computer, and the intensity of ttuorescence signal and its ratio were calculated. Results： A total of 92 genes were screened out and its expression difference was more than 2 times in the peripheral blood between the patients with esophageal carcinoma and normal subjects. Among these, the expression difference of 36 genes was more than 3 times. Two human urokinase plasminogen activator surface receptor （UPAR） genes, 80K-L protein gene, human protein tyrosine-phosphatase gent arid proto-oncogene protein mRNA were significantly up-regulated, while the collagen V type （α-2 gene was markedly down-regulated. Conclusion： 80K-L protein gene, tyrosinephophatase gene, proto-oncogene protein arid the collagen V type α-2 gene might be associated with the ontogenesis, development and its metastasis in the esophageal carcinoma. The UPAR gene may play important roles in the diagnosing the micrometastasis in the peripheral blood of esophageal carcinoma.展开更多
[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated f...[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated from pig, and precursor dendritic cells were obtained by adherence method. The dendritic cells were treated by recombinant porcine granulocyte-monocyte colony stimulating factor （rpGM-CSF） and recombinant porcine interleukin-4 （rplL-4） together, and lipopolysaccharide （LPS） respectively. The cells in different time periods were collected. The morphology of the collected cells was observed by scanning electron microscopy; the expression of surface molecules and phagocytic ability to FITC-dextran were detected by flow cy- tometry; and the stimulating ability for allogeneic T cells was detected by mixed lymphocyte reaction. [Result] The DCs suffering maturation induction in vitro showed typical dendritic morphology; compared with those of DCs untreated by LPS, the cell surface expression of CDla, CD80, CD86, SLAII and CD172a of DCs treated by LPS was significantly increased, the phagocytic ability was reduced slightly, and the stimulating ability for allogeneic T cells was enhanced to some extent. [Conclusion] An in vitro culture method was successfully established for porcine MoDCs in this study, laying a foundation for further study on the role of porcine MoDCs in immunoregulation and anti-virus infection.展开更多
AIM： To detect the expression of CD44 correlated with the ability of micro-metastasis in peripheral blood and bone marrow of patients with gastric cancer and to deduce its clinical significance. METHODS： Preoperativ...AIM： To detect the expression of CD44 correlated with the ability of micro-metastasis in peripheral blood and bone marrow of patients with gastric cancer and to deduce its clinical significance. METHODS： Preoperative peripheral blood and bone marrow specimens from 46 patients with gastric cancer and 6 controls were studied by semi-quantitative RTPCR amplification of CD44v6mRNA. Preoperative and postoperative peripheral blood specimens from 40 patients with gastric cancer and 14 controls were studied by quantitative RT-PCR amplification of CD44v6mRNA in the corresponding period. RESULTS： Semi-quantitative RT-PCR amplification showed that CD44v6mRNA expression of peripheral blood and bone marrow was positive in 39 （84.8%） and 40 （86.9%） of 46 patients with gastric cancer, respectively. In peripheral blood, CD44v6mRNA expression was positive for diffuse type in 30 （93.8%） of 32 patients and for intestinal type in 9 （64.3%） of 14 patients. On the other hand, in bone marrow, CD44v6mRNA expression was positive for diffuse type in 31 （96.9%） of 32 patients and for intestinal type in 10 （71.4%） of 14 patients. There was a significant difference between the diffuse type and intestinal type. Quantitative RTopCR amplification demonstrated that CD44v6mRNA was not expressed in the peripheral blood of controls and CD44v6mRNA expression was positive for preoperative peripheral blood in 40 patients with gastric cancer, the expression levels being from 4.9 × 10^2 to 3.2× 10^5 copies/g RNA. The average expression level of CD44v6mRNA in peripheral blood was 3.9 × 10^10 copies/g RNA. The expression levels of CD44v6mRNA in peripheral blood in gastric cancer patients after curative operation increased from 5.5 × 100 to 7.6 × 10 copies/g RNA （P = 0.00496）. After curative operation, the expression level decreased markedly. CONCLUSION： Semi-quantitative and quantitative RTPCR amplification for CD44v6mRNA is a sensitive and specific method for the detection of micro-metastasis in peripheral blood and bone marrow, which might be used as an indicator of tumor burden and therapeutic effect.展开更多
BACKGROUND: Interleukin-18 (IL-18), a pro-inflamma- tory cytokine that induces interferon-γ (IFN-γ) production in T cells and natural killer cells, plays a critical role in the T-lymphocyte helper type 1 ( Th1) resp...BACKGROUND: Interleukin-18 (IL-18), a pro-inflamma- tory cytokine that induces interferon-γ (IFN-γ) production in T cells and natural killer cells, plays a critical role in the T-lymphocyte helper type 1 ( Th1) response. This study was designed to explore the effect of IL-18 on peripheral blood mononuclear cells ( PBMCs) derived from chronic hepatitis B (CHB) and on hepatitis B virus (HBV) DNA released by HepG2.2.15 cell lines, which were transfected with hepatitis B virus gene in vitro. METHODS: PBMCs isolated from 25 healthy people and 25 patients with CHB were stimulated with HBcAg and IL-18 of various concentrations for 72 hours. The levels of IFN-γ in the supernatants of cultured PBMCs were determined by ELISA. After the stimulation of IL-18 of various concentra- tions, PBMCs derived from one patient were co-cultured for 96 hours with HepG2. 2. 15 cells which had been cul- tured for 24 hours, and then the supernatants were collected by centrifugation and used for HBV DNA quantitative as- say. RESULTS: When PBMCs were stimulated by HBcAg and IL-18 at various concentrations, the levels of IFN-γ in the supernatants of CHB groups were much higher than those in normal control groups, at 0.2 ng/ml: t =11.70, P< 0.01; at 1.0 ng/ml: t =16.19, P<0.01; and at5.0 ng/ml: t =20.12, P <0.01. In the CHB groups, the levels of IFN-γ in the supernatants of PBMCs stimulated by HBcAg alone were lower than both those stimulated by HBcAg and EL-18 at various concentrations and those stimulated by HBcAg and EL-18 (5.0 ng/ml) together with EL-12 (mild: t = 2.20, P<0.05; moderate; t=2.97, P<0.05; severe; t = 0.66, P >0.05). The content of HBV DNA in the superna- tant of co-cultivation of HepG2. 2. 15 cells and PBMCs without stimulated materials was higher than that stimula-ted by HBcAg and EL-18 at various concentrations of HBc- Ag and IL-18 together with IL-12/IFN-α1lb. CONCLUSION: DL-18 can induce IFN-γ secretion and pro- bably play a key role in the modulation of both innate and adaptive immunity. It has implications in improving im- munoregulatory effect and increasing the ability of immune cells to kill cells infected by virus.展开更多
BACKGROUND: Post-hepatitic cirrhosis is regarded as common and severe form of liver damage. Interferon gamma-inducible protein 10 (IP-10), a member of the non-ELR (glutamic-leucine-arginine) motif CXC chemokine family...BACKGROUND: Post-hepatitic cirrhosis is regarded as common and severe form of liver damage. Interferon gamma-inducible protein 10 (IP-10), a member of the non-ELR (glutamic-leucine-arginine) motif CXC chemokine family, has recently been shown to recruit and activate specific subsets of leukocytes to sites of inflammation or an immune response during the development of hepatic cirrhosis. However, the effects of IP-10 and IP-10 mRNA on inflammatory infiltration at local sites and in the peripheral blood of patients with post-hepatitic cirrhosis as well as their relationship with viral load are still poorly defined. This study aimed to detect the relationship between the expression of IP-10 in serum, IP-10 mRNA in peripheral blood mononuclear cells (PBMCs), and the levels of HBV DNA in the serum of patients, and to explore their role in the pathogenesis of cirrhosis. METHODS: Typical patients with cirrhosis after HBV infection were selected, and their serum IP-10 concentrations were evaluated with ELISA, the content of IP-10 mRNA in PBMCs was measured by real-time PCR, and the load of HBV DNA in serum and PBMCs was assessed by semi-quantitative analysis of gel imaging. RESULTS: The levels of IP-10 in serum and IP-10 mRNA in PBMCs of patients with cirrhosis were 299.9 +/- 77.2 pg/ml and 0.7500 +/- 0.1495, respectively. They were higher than those of controls (P<0.05) and also increased in the HBV DNA(+) groups (P<0.05, P<0.01) to 343.0 +/- 80.3 pg/ml and 0.8465 +/- 0.1528, respectively. The levels of IP-10 in serum and IP-10 mRNA in PBMCs were clearly correlated with the load of HBV DNA (P<0.01). CONCLUSIONS: The levels of IP-10 and IP-10 mRNA in the peripheral blood of patients with cirrhosis increase are closely correlated with the load of HBV DNA in serum, and play a key role in the progression of post-hepatitic cirrhosis. (Hepatobiliary Pancreat Dis Int 2010; 9: 280-286)展开更多
AIM： To investigate the dynamic alteration of telomerase expression during development of hepatocellular carcinoma （HCC） and its diagnostic implications in liver tissues or peripheral blood mononuclear cells for HC...AIM： To investigate the dynamic alteration of telomerase expression during development of hepatocellular carcinoma （HCC） and its diagnostic implications in liver tissues or peripheral blood mononuclear cells for HCC. METHODS： Dynamic expressions of liver telomerase during malignant transformation of hepatocytes were observed in Sprague-Dawly （SD） rats fed with 0.05% of 2-fluoenyacetamide （2-FAA）. Total RNA and telomerase were extracted from rat or human liver tissues. The telomerase activities in livers and in circulating blood were detected by a telomeric repeat amplification protocol-enzyme-linked immunosorbent assay （TRAP- ELISA）, and its diagnostic value was investigated in patients with benign or malignant liver diseases. RESULTS： The hepatoma model displayed the dynamic expression of hepatic telomerase during HCC development. The telomerase activities were consistent with liver total RNA levels （r = 0.83, P 〈 0.01） at the stages of degeneration, precancerosis, and cancerization of hepatocytes. In HCC patients, the telomerase levels in HCC tissues were significantly higher than in their adjacent non-cancerous tissues, but liver total RNA levels were lower in the former than in the latter. Although the circulating telomerase of HCC patients was abnormally expressed among patients with chronic liver diseases, the telomerase activity was a non-specific marker for HCC diagnosis, because the incidence was 15.7% in normal control, 25% in chronic hepatitis, 45.9% in liver cirrhosis, and 85.2% in HCC, respectively when absorbance value of telomerase activity was more than 0.2. If the value was over 0.6, the incidence was 60% in HCC group and 0% in any of the others （P 〈 0.01） except in two cases with liver cirrhosis. However, the combination of circulating telomerase with serum alpha-fetoprotein level could increase the positive rate and the accuracy （92.6%, 125 of 135） of HCC diagnosis. CONCLUSION： The overexpression of telomerase is associated with HCC development, and its abnormality in liver tissues or in peripheral blood could be a useful marker for diagnosis and prognosis of HCC.展开更多
The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible role...The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.展开更多
AIM： To study the dynamic changes of hepatits B virus （HBV） DNA in serum and peripheral blood mononuclear cells （PBMCs） of patients after lamivudine therapy. METHODS： A total of 72 patients with chronic HBV infe...AIM： To study the dynamic changes of hepatits B virus （HBV） DNA in serum and peripheral blood mononuclear cells （PBMCs） of patients after lamivudine therapy. METHODS： A total of 72 patients with chronic HBV infection were included in this study. All patients were confirmed to have the following conditions： above 16 years of age, elevated serum alanine amonotransferase （ALT）, positive hepatitis B e antigen （HBeAg）, positive HBV DNA in serum and PBMCs, negative antibodies against HAV, HCV, HDV, HEV. Other possible causes of chronic liver damages, such as drugs, alcohol and autoimmune diseases were excluded. Seventy-two cases were randomly divided into lamivudine treatment group （n = 42） and control group （n = 30）. HBV DNA was detected both in serum and in PBMCs by fluorescence quantitative polymerase chain reaction （PCR）, during and after lamivudine treatment. RESULTS： In the treatment group, HBV DNA became negative both in serum and in PBMC, of, 38 and 25 out of 42 cases respectively during the 48 wk oflamivudine treatment, the negative rate was 90.5% and 59.5% respectively. In the control group, the negative rate was 23.3% and 16.7% respectively. It was statistically significant at 12, 24 and 48 wk as compared with the control group （P 〈 0.005）. The average conversion period of HBV DNA was 6 wk （2-8 wk） in serum and 16 wk （8-24 wk） in PBMC.CONCLUSION： Lamivudine has remarkable effects on HBV replication both in serum and The inhibitory effect on HBV DNA in PBMCs than that in serum inhibitory in PBMCs. is weaker展开更多
Astragalus membranaceus(A.membranaceus)is a widely used traditional herb in China and Korea.A.membranaceus polysaccharides(AMP),which make up a major part of the root extract,have been shown to modulate immune modulat...Astragalus membranaceus(A.membranaceus)is a widely used traditional herb in China and Korea.A.membranaceus polysaccharides(AMP),which make up a major part of the root extract,have been shown to modulate immune modulations,especially activation of bone marrow-derived dendritic cells(BMDCs)and T cells.However,the immune stimulatory effect of AMP in the mouse in vivo and human peripheral blood DCs(PBDCs)has not been well investigated.In this study,we found that intravenous(i.v.)injection of AMP in C57 BL/6 mice induced remarkable elevations in co-stimulatory and MHC class I and II molecule levels in the splenic DCs and its subsets.The stimulatory effect of DCs by AMP was elevated 6 h after treatment,which rapidly decreased 18 h after injection.Furthermore,AMP promoted intracellular production of pro-inflammatory cytokines in spleen DC subsets,which contributed elevation of serum cytokine levels.Finally,the AMP promoted PBDC activation.Thus,these results demonstrate that AMP can be used as an immune stimulatory molecules in human and mouse.展开更多
The aim of the present study was to investigate the expression of toll-like receptors （TLR） 9 in peripheral blood mononuclear cells （PBMC） of patients with chronic hepatitis B and C with different virus copies. Th...The aim of the present study was to investigate the expression of toll-like receptors （TLR） 9 in peripheral blood mononuclear cells （PBMC） of patients with chronic hepatitis B and C with different virus copies. The study group included 90 patients （60 with chronic hepatitis B, and 30 with chronic hepatitis C）, and 20 healthy people served as control group. The protein and mRNA levels of TLR9 were detected by using flow cytometry and real-time PCR. The serum viral copies of HBV and HCV were measured in all patients, and the correlation between HBV-DNA copies or HCV-RNA copies and the TLR9 expression was analyzed. Our results demonstrated that HBV or HCV infection led to a decreased expression of TLR9 mRNA and protein compared to the control group （P〈0.05）. The TLR9 protein and mRNA levels were negatively correlated with serum viral copies of HBV and HCV （r=-0.632, r=-0.909, P〈0.01）. It was concluded that TLR9 mRNA and protein are down-regulated in PBMC of HBV-infected or HCV-infected patients, and they are negatively correlated with serum viral copies and play an important role in detecting viral replication of HBV and HCV.展开更多
基金supported by Hunan Provincial Key Research and Development Program,No.2021SK2002(to BW)the Natural Science Foundation of Hunan Province of China(General Program),No.2021JJ30938(to YL)。
文摘Degenerative cervical myelopathy is a common cause of spinal cord injury,with longer symptom duration and higher myelopathy severity indicating a worse prognosis.While numerous studies have investigated serological biomarkers for acute spinal cord injury,few studies have explored such biomarkers for diagnosing degenerative cervical myelopathy.This study involved 30 patients with degenerative cervical myelopathy(51.3±7.3 years old,12 women and 18 men),seven healthy controls(25.7±1.7 years old,one woman and six men),and nine patients with cervical spondylotic radiculopathy(51.9±8.6 years old,three women and six men).Analysis of blood samples from the three groups showed clear differences in transcriptomic characteristics.Enrichment analysis identified 128 differentially expressed genes that were enriched in patients with neurological disabilities.Using least absolute shrinkage and selection operator analysis,we constructed a five-gene model(TBCD,TPM2,PNKD,EIF4G2,and AP5Z1)to diagnose degenerative cervical myelopathy with an accuracy of 93.5%.One-gene models(TCAP and SDHA)identified mild and severe degenerative cervical myelopathy with accuracies of 83.3%and 76.7%,respectively.Signatures of two immune cell types(memory B cells and memory-activated CD4^(+)T cells)predicted levels of lesions in degenerative cervical myelopathy with 80%accuracy.Our results suggest that peripheral blood RNA biomarkers could be used to predict lesion severity in degenerative cervical myelopathy.
文摘Parkinson’s disease(PD)is a common neurodegenerative disorder with profound impact on patients’quality of life and long-term health,and early detection and intervention are particularly critical.In recent years,the search for precise and reliable biomarkers has become one of the key strategies to effectively address the clinical challenges of PD.In this paper,we systematically evaluated potential biomarkers,including proteins,metabolites,epigenetic markers,and exosomes,in the peripheral blood of PD patients.Protein markers are one of the main directions of biomarker research in PD.In particular,α‑synuclein and its phosphorylated form play a key role in the pathological process of PD.It has been shown that aggregation ofα-synuclein may be associated with pathologic protein deposition in PD and may be a potential marker for early diagnosis of PD.In terms of metabolites,uric acid,as a metabolite,plays an important role in oxidative stress and neuroprotection in PD.It has been found that changes in uric acid levels may be associated with the onset and progression of PD,showing its potential as an early diagnostic marker.Epigenetic markers,such as DNA methylation modifications and miRNAs,have also attracted much attention in Parkinson’s disease research.Changes in these markers may affect the expression of PD-related genes and have an important impact on the onset and progression of the disease,providing new research perspectives for the early diagnosis of PD.In addition,exosomes,as a potential biomarker carrier for PD,are able to carry a variety of biomolecules involved in intercellular communication and pathological regulation.Studies have shown that exosomes may play an important role in the pathogenesis of PD,and their detection in blood may provide a new breakthrough for early diagnosis.It has been shown that exosomes may play an important role in the pathogenesis of PD,and their detection in blood may provide new breakthroughs in early diagnosis.In summary,through in-depth evaluation of biomarkers in the peripheral blood of PD patients,this paper demonstrates the important potential of these markers in the early diagnosis of PD and in the study of pathological mechanisms.Future studies will continue to explore the clinical application value of these biomarkers to promote the early detection of PD and individualized treatment strategies.
基金Supported by the National Natural Science Foundation of China,No.82300857.
文摘BACKGROUND Patients with acute-on-chronic liver failure(ACLF)experience severe immune dysfunction.Liver transplantation(LT)significantly improves survival outcomes.However,the characteristics of peripheral blood lymphocyte subsets(PBLSs)in this patient population are not well defined,and the dynamics of immune reconstitution post-LT are insufficiently understood.AIM To characterize PBLSs in patients with ACLF prior to LT and to evaluate PBLS reconstitution after LT.METHODS Clinical data from patients undergoing LT in the Transplantation Center,The Third Xiangya Hospital from January 2022 to December 2023 were analyzed retrospectively.Our cohort comprised 44 patients with ACLF,16 patients with acute decompensation of cirrhosis,and 23 patients with compensated cirrhosis.Twenty healthy volunteers were included as controls.PBLSs were evaluated across all groups.The relationship between PBLSs and post-LT prognosis was assessed,and dynamic changes in PBLSs among patients with ACLF were analyzed at different time points.RESULTS Patients with ACLF exhibited a marked reduction in PBLSs compared with healthy volunteers.Natural killer(NK)cell counts were further reduced in patients with ACLF when compared with patients with compensated cirrhosis.PBLSs did not correlate with the etiology or severity of ACLF or with established liver failure scores.Following LT,a rapid restoration of NK cells and B cells was observed in patients with ACLF.However,the cluster of differentiation(CD)3+T cell and CD4+T cell counts decreased 14 days post-LT and subsequently returned to preoperative levels by day 21.CONCLUSION Patients with ACLF exhibited markedly reduced PBLSs,with decreased NK cells potentially linked to progression from compensated cirrhosis to liver failure.NK and B cell were rapidly restored after LT.
文摘In the context of global aging,mild behavioral impairment(MBI)is present in 48.9%of patients with mild cognitive impairment(MCI).MBI,a neurobehavioral syndrome in the elderly,is an independent risk factor for cognitive decline and is closely related to peripheral blood biomarkers associated with Alzheimer's disease,offering new diagnostic and interventional avenues for early MCI.To summarize evidence on peripheral blood biomarkers related to MBI and their underlying mechanisms involving neuroinflammation,tau pathology,and oxidative stress,a systematic review of studies published between 2015 and 2024 was conducted.MBI is closely associated with peripheral blood biomarker changes.Neuroinflammatory markers like glial fibrillary acidic protein and neurofilament light indicate astrocyte activation and neural circuit disruption,with glial fibrillary acidic protein levels correlating with impulse dyscontrol scores.Chitinase-3-like protein 1,a marker of blood-brain barrier integrity,exacerbates neuroinflammation and is linked to depressive symptoms and hippocampal atrophy.Elevated phosphorylated tau proteins in blood correlate with brain tau deposition,increasing the risk of MBI and impairing cognition.Oxidative stress markers damage neurons and disrupt neurotransmission,and concurrent alterations in malondialdehyde and superoxide dismutase levels significantly elevate the risk of MBI.The correlation between MBI and biomarkers offers new diagnostic and interventional directions for early MCI.Future research should standardize MBI assessment,conduct longitudinal studies,explore biomarker-MBI relationships,investigate psychosocial impacts,and develop advanced detection methods.
基金supported by Russian Science Foundation,RSF 23-75-00009(part of the study corresponding to finding 1)Part of the study corresponding to finding 2 was carried out within the state assignment,FSR No.:122020300045-5(03.02.2022).
文摘Objectives To compare respiratory parameters of peripheral blood mononuclear cell mitochondria and iron metabolism indicators in patients with different NYHA functional classes of ischemic heart failure(HF).Methods This single center, prospective, non-blinded study enrolled 20 patients with diagnosed chronic HF of ischemic genesis with reduced and mildly reduced left ventricle ejection fraction. The maximum oxygen consumption at the peak of the exercise test(VO2peak), iron metabolism parameters and respiratory activity of peripheral blood mononuclear cell mitochondria were assessed.Results Among the patients, a half of individuals were diagnosed with iron deficiency. Subgroups of patients with different HF severity did not significant differ in VO2peak(P=0.209), serum iron(P=0.468) and ferritin(P=0.235) levels. But there was a trend in increasing in these parameters with increasing NYHA HF functional class. Respiratory control coefficient(RC) in NADdependent and FAD-dependent mitochondrial oxidation were lower in patients with NYHA HF Ⅲ functional class compared to individuals with NYHA HF I functional class(P=0.028 and P=0.040, respectively). Serum iron(P=0.026), ferritin(P=0.045)levels, transferrin saturation(P=0.006) were negatively correlated with RC in NAD-dependent mitochondrial oxidation.Conclusions In aggravation of ischemic HF NYHA FC, there is a decrease in RC of PBMC mitochondria during the oxidation of NAD-dependent and FAD-dependent substrates. In the whole sample, patients with laboratory-confirmed iron deficiency accounted a half of the total number. Iron metabolism parameters had a paradoxical inverse relationship with the level of RC in PBMC mitochondria of patients with HF.
基金Project of Guangdong Provincial Medical Science and Technology Research Fund(A2022011)Major Science and Technology Project of Shenzhen Nanshan District Health System(NSZD2023067)Sub-project of Education(Health)Science and Technology Project of Nanshan District Technology Research and Development and Creative Design Project in Shenzhen(NS2022002)。
文摘Objective:To explore the accuracy and stability of the results of peripheral blood routine tests at different time points after anticoagulation and standing,providing a scientific basis for actual clinical work.Methods:In this study,30 patients who visited the hospital in October 2023 were randomly selected and divided into two groups(15 cases in each group).The same collection method was used for routine blood tests.The tests were performed after anticoagulation and standing for 5 minutes,1 hour,and 5 minutes,2 hours respectively,and the routine blood test indicators at different time points were compared.Results:After comparison,there were no significant differences in the results of routine blood tests at 5 minutes after mixing,anticoagulating,and standing peripheral blood and those at 1 hour and 2 hours(P>0.05).Conclusion:The results of peripheral blood after mixing,anticoagulating,and standing for 5 minutes are stable compared with those after standing for 1 hour and 2 hours.In actual work,the pre-test turnaround time can be appropriately extended.
基金Supported by the Sichuan Provincial Science and Technology Support Project(No.2024YFFK0303).
文摘AIM:To investigate the possible relationship between inflammatory biomarkers in the peripheral blood of patients with branch retinal vein occlusion(BRVO).METHODS:A total of 63 BRVO patients were enrolled in this cross-sectional observational study.Meanwhile,63 age-and gender-matched cataract patients were included as controls.Complete blood count and biochemical tests were performed,and inflammatory biomarkers including platelet to lymphocyte ratio(PLR),red blood cell distribution width to albumin ratio(RAR),neutrophil to lymphocyte ratio(NLR),systemic immune inflammation index(SII),and monocyte to high density lipoprotein cholesterol ratio(MHR)were compared between the two groups.RESULTS:There were no significant differences between the two groups in terms of age,sex,and prevalence of diabetes mellitus.Compared with the controls,patients with BRVO had a higher prevalence of hypertension and higher body mass index(BMI).Red blood cell distribution width(RDW),triglycerides,MHR,NLR,and RAR were elevated,whereas lymphocyte count and high-density lipoprotein were decreased in the BRVO group.Multivariate logistic regression analysis revealed that NLR(adjusted OR=1.686,95%CI 1.075-2.646),RAR(adjusted OR=8.930,95%CI 1.911-41.730),and body mass index(BMI;adjusted OR=1.174,95%CI 1.010-1.365)were significantly associated with the risk of BRVO.In the receiver operating characteristic analysis,the area under the curve for NLR,RAR,and BMI were 0.602,0.630,and 0.603,respectively.The sensitivity and specificity were 61.9%and 60.3%,38.1%and 82.5%,and 61.9%and 57.1%,respectively.CONCLUSION:Peripheral blood inflammatory biomarkers are elevated in BRVO patients,suggesting systemic inflammation involvement.NLR,RAR,and BMI are positively correlated with BRVO.Monitoring NLR and RAR and strict weight control may be beneficial for the prevention and treatment of BRVO.
基金This study was supported by grants from the Key Project of the Chinese Ministry of Science and Technology(2017ZX102022022)National Key Research and Development Program of China(2021YFC2301801).
文摘Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnostic value for hepatitis B virus-associated hepatocellular carcinoma(HBV-HCC). Methods: We enrolled 197 patients, 111 with HBV-HCC, 51 with chronic hepatitis B(CHB), and 35 healthy controls(HCs). The methylation status of GLDC promoter in peripheral mononuclear cells(PBMCs) was identified by methylation specific polymerase chain reaction(MSP). The mRNA expression was examined using real-time quantitative polymerase chain reaction(q PCR). Results: The methylation frequency of the GLDC promoter was significantly lower in HBV-HCC patients(27.0%) compared to that in CHB patients(68.6%) and HCs(74.3%)( P < 0.001). The methylated group had lower alanine aminotransferase level( P = 0.035) and lower rates of tumor node metastasis(TNM) Ⅲ/Ⅳ( P = 0.043) and T3/T4( P = 0.026). TNM stage was identified to be an independent factor for GLDC promoter methylation. GLDC mRNA levels in CHB patients and HCs were significantly lower than those in HBV-HCC patients( P = 0.022 and P < 0.001, respectively). GLDC mRNA levels were significantly higher in HBV-HCC patients with unmethylated GLDC promoters than those with methylated GLDC promoters( P = 0.003). The diagnostic accuracy of alpha-fetoprotein(AFP) combined with GLDC promoter methylation for HBV-HCC was improved compared with that of AFP alone(AUC: 0.782 vs. 0.630, P < 0.001). In addition, GLDC promoter methylation was an independent predictor for overall survival of HBV-HCC patients( P = 0.038). Conclusions: The methylation frequency of GLDC promoter was lower in PBMCs from HBV-HCC patients than that from patients with CHB and HCs. The combination of AFP and GLDC promoter hypomethylation significantly improved the diagnostic accuracy of HBV-HCC.
基金supported by the National Key Research and Development Project of China(No.2022YFD2400401)the Key Research and Development Project of Shandong(No.2021LZGC028)the Academician Special Program of Shandong Province(No.2023ZLYS02)。
文摘As an efficient immunostimulant,chitosan oligosaccharide(COS)can enhance the immunity of teleosts;however,the underlying molecular mechanisms still require elucidation.Competing endogenous RNAs(ceRNAs)are vital regulators in the immune response,but their roles in half-smooth tongue sole(Cynoglossus semilaevis)remain unclear.In this study,for the first time,we studied whole-transcriptome expression profiles and analyzed ceRNA networks in peripheral blood leukocytes of half-smooth tongue sole treated with COS.A total of 19 circRNAs(DE-circRNAs),18 miRNAs(DE-miRNAs)and 50 previously identified lncRNAs(DElncRNAs)were differentially expressed after COS stimulation.The DE-lncRNAs and DE-miRNAs targeted numerous immunity-related genes,and were enriched in important pathways,including MAPK and Toll-like receptor signaling pathways,suggesting the immunoregulatory roles of COS.Furthermore,we constructed circRNA-miRNA-mRNA and lncRNA-miRNA-mRNA regulatory networks using DE-circRNAs,DE-miRNAs,DE-lncRNAs,and DE-mRNAs.Additionally,a ceRNA network with immunity-related DEmRNAs was constructed,showing that 3 DE-circRNAs,12 DE-lncRNAs,and 29 DEGs exhibited crosstalk through 9 DE-miRNAs.Intriguingly,a DE-miRNA in the ceRNA network,miR-144-3p,was targeted by DE-lncRNA tnrc6a,and negatively regulated the genes of inhibitor of nuclear factor kappa B kinases(IKKs)(ikbkg,ikbkb,and ikbip)and c3ar1.Ikbkg,ikbkb,and c3ar1 were significantly up-regulated in macrophages stimulated by LPS.It could be inferred that ncRNAs participated in the immune and inflammatory response by acting as ceRNAs after COS stimulation in teleosts.These findings indicate that COS could enhance the immunity of teleosts by regulating ncRNAs,and lay the foundation for further practical application of COS in aquaculture.
文摘BACKGROUND Gastric cancer is one of the most common malignant tumors worldwide,and surgical resection is one of the main ways to treat gastric cancer.However,the immune status of postoperative patients is crucial for prognosis and survival,and immune cells play an important role in this process.Therefore,it is helpful to understand the immune status of postoperative patients by evaluating the levels of peripheral blood immune cells,especially total T cells(CD3+),helper T cells(CD3+CD4+),and suppressor T cells(CD3+CD8+),and its relationship to sur-vival.AIM To analyzed the immune cells in peripheral blood of patients with gastric cancer after surgery,detect the levels of total T cells,helper T cells and suppressor T cells.METHODS A total of 58 patients with gastric cancer who received surgical treatment were included in the retrospective study.Flow cytometry was used to detect the level of peripheral blood immune cells and analyze the correlation between total T cells,helper T cells and inhibitory T cells.To explore the relationship between these immune markers and patient survival.RESULTS The results showed that the levels of total T cells,helper T cells,and suppressor T cells changed in patients after gastric cancer surgery.There was a significant positive correlation between total T cells,helper T cells and suppressor T cells(r=0.35,P<0.01;r=0.56,P<0.01).However,there was a negative correlation between helper T cells and suppressor T cells(r=-0.63,P<0.01).Follow-up showed that the survival rate of patients in the high-level total T cell group was significantly higher than that in the low-level group(28.87±24.98 months vs 18.42±16.21 months).The survival curve shows that the curve of patients in the high-level group is shifted to the upper right,and that of the low-level group is shifted downward.There was no significant difference between the levels of helper T cells and suppressor T cells and patient survival time.CONCLUSION By detecting peripheral blood immune cells with flow cytometry,we can initially evaluate the immune status of patients after gastric cancer surgery and initially explore its relationship with patient survival.
基金This project was supported by a grant from the Zhejiang Medical and Health Science Foundation (No. 2002A023).
文摘Objective： To study the genes expression profile differences in the peripheral blood between esophageal carcinoma patients and normal subjects using the gene chip technique and screen out the esophageal early conceration associated genes. Methods： The total RNA was extracted and purified in the peripheral blood obtained from the patients with esophageal carcinoma and normal subjects. The first strand of cDNA was synthesized through retro-transcription and labeled with Cy5 and Cy3 fluorescence as probes. The mixed probes were hybridized with a piece of 4096 double dot human whole gene chip. The acquired image was analyzed by microarrav suite software using a digital computer, and the intensity of ttuorescence signal and its ratio were calculated. Results： A total of 92 genes were screened out and its expression difference was more than 2 times in the peripheral blood between the patients with esophageal carcinoma and normal subjects. Among these, the expression difference of 36 genes was more than 3 times. Two human urokinase plasminogen activator surface receptor （UPAR） genes, 80K-L protein gene, human protein tyrosine-phosphatase gent arid proto-oncogene protein mRNA were significantly up-regulated, while the collagen V type （α-2 gene was markedly down-regulated. Conclusion： 80K-L protein gene, tyrosinephophatase gene, proto-oncogene protein arid the collagen V type α-2 gene might be associated with the ontogenesis, development and its metastasis in the esophageal carcinoma. The UPAR gene may play important roles in the diagnosing the micrometastasis in the peripheral blood of esophageal carcinoma.
基金Supported by Fundamental and Advanced Research Projects of Henan Province(152300410076,2015-2017)Key Science and Technology Program of Henan Province(152102110048,2015-2017)~~
文摘[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated from pig, and precursor dendritic cells were obtained by adherence method. The dendritic cells were treated by recombinant porcine granulocyte-monocyte colony stimulating factor （rpGM-CSF） and recombinant porcine interleukin-4 （rplL-4） together, and lipopolysaccharide （LPS） respectively. The cells in different time periods were collected. The morphology of the collected cells was observed by scanning electron microscopy; the expression of surface molecules and phagocytic ability to FITC-dextran were detected by flow cy- tometry; and the stimulating ability for allogeneic T cells was detected by mixed lymphocyte reaction. [Result] The DCs suffering maturation induction in vitro showed typical dendritic morphology; compared with those of DCs untreated by LPS, the cell surface expression of CDla, CD80, CD86, SLAII and CD172a of DCs treated by LPS was significantly increased, the phagocytic ability was reduced slightly, and the stimulating ability for allogeneic T cells was enhanced to some extent. [Conclusion] An in vitro culture method was successfully established for porcine MoDCs in this study, laying a foundation for further study on the role of porcine MoDCs in immunoregulation and anti-virus infection.
基金Supported by the grant from Science and Technology Committee of Jiangsu Province,No.457-99064
文摘AIM： To detect the expression of CD44 correlated with the ability of micro-metastasis in peripheral blood and bone marrow of patients with gastric cancer and to deduce its clinical significance. METHODS： Preoperative peripheral blood and bone marrow specimens from 46 patients with gastric cancer and 6 controls were studied by semi-quantitative RTPCR amplification of CD44v6mRNA. Preoperative and postoperative peripheral blood specimens from 40 patients with gastric cancer and 14 controls were studied by quantitative RT-PCR amplification of CD44v6mRNA in the corresponding period. RESULTS： Semi-quantitative RT-PCR amplification showed that CD44v6mRNA expression of peripheral blood and bone marrow was positive in 39 （84.8%） and 40 （86.9%） of 46 patients with gastric cancer, respectively. In peripheral blood, CD44v6mRNA expression was positive for diffuse type in 30 （93.8%） of 32 patients and for intestinal type in 9 （64.3%） of 14 patients. On the other hand, in bone marrow, CD44v6mRNA expression was positive for diffuse type in 31 （96.9%） of 32 patients and for intestinal type in 10 （71.4%） of 14 patients. There was a significant difference between the diffuse type and intestinal type. Quantitative RTopCR amplification demonstrated that CD44v6mRNA was not expressed in the peripheral blood of controls and CD44v6mRNA expression was positive for preoperative peripheral blood in 40 patients with gastric cancer, the expression levels being from 4.9 × 10^2 to 3.2× 10^5 copies/g RNA. The average expression level of CD44v6mRNA in peripheral blood was 3.9 × 10^10 copies/g RNA. The expression levels of CD44v6mRNA in peripheral blood in gastric cancer patients after curative operation increased from 5.5 × 100 to 7.6 × 10 copies/g RNA （P = 0.00496）. After curative operation, the expression level decreased markedly. CONCLUSION： Semi-quantitative and quantitative RTPCR amplification for CD44v6mRNA is a sensitive and specific method for the detection of micro-metastasis in peripheral blood and bone marrow, which might be used as an indicator of tumor burden and therapeutic effect.
文摘BACKGROUND: Interleukin-18 (IL-18), a pro-inflamma- tory cytokine that induces interferon-γ (IFN-γ) production in T cells and natural killer cells, plays a critical role in the T-lymphocyte helper type 1 ( Th1) response. This study was designed to explore the effect of IL-18 on peripheral blood mononuclear cells ( PBMCs) derived from chronic hepatitis B (CHB) and on hepatitis B virus (HBV) DNA released by HepG2.2.15 cell lines, which were transfected with hepatitis B virus gene in vitro. METHODS: PBMCs isolated from 25 healthy people and 25 patients with CHB were stimulated with HBcAg and IL-18 of various concentrations for 72 hours. The levels of IFN-γ in the supernatants of cultured PBMCs were determined by ELISA. After the stimulation of IL-18 of various concentra- tions, PBMCs derived from one patient were co-cultured for 96 hours with HepG2. 2. 15 cells which had been cul- tured for 24 hours, and then the supernatants were collected by centrifugation and used for HBV DNA quantitative as- say. RESULTS: When PBMCs were stimulated by HBcAg and IL-18 at various concentrations, the levels of IFN-γ in the supernatants of CHB groups were much higher than those in normal control groups, at 0.2 ng/ml: t =11.70, P< 0.01; at 1.0 ng/ml: t =16.19, P<0.01; and at5.0 ng/ml: t =20.12, P <0.01. In the CHB groups, the levels of IFN-γ in the supernatants of PBMCs stimulated by HBcAg alone were lower than both those stimulated by HBcAg and EL-18 at various concentrations and those stimulated by HBcAg and EL-18 (5.0 ng/ml) together with EL-12 (mild: t = 2.20, P<0.05; moderate; t=2.97, P<0.05; severe; t = 0.66, P >0.05). The content of HBV DNA in the superna- tant of co-cultivation of HepG2. 2. 15 cells and PBMCs without stimulated materials was higher than that stimula-ted by HBcAg and EL-18 at various concentrations of HBc- Ag and IL-18 together with IL-12/IFN-α1lb. CONCLUSION: DL-18 can induce IFN-γ secretion and pro- bably play a key role in the modulation of both innate and adaptive immunity. It has implications in improving im- munoregulatory effect and increasing the ability of immune cells to kill cells infected by virus.
基金supported by grants from the Natural Science Foundation of Anhui Province(090413138)the Natural Science Foundation of the Department of Education of Anhui Province(KJ2007A019,KJ2009A032,KJ2010A086)
文摘BACKGROUND: Post-hepatitic cirrhosis is regarded as common and severe form of liver damage. Interferon gamma-inducible protein 10 (IP-10), a member of the non-ELR (glutamic-leucine-arginine) motif CXC chemokine family, has recently been shown to recruit and activate specific subsets of leukocytes to sites of inflammation or an immune response during the development of hepatic cirrhosis. However, the effects of IP-10 and IP-10 mRNA on inflammatory infiltration at local sites and in the peripheral blood of patients with post-hepatitic cirrhosis as well as their relationship with viral load are still poorly defined. This study aimed to detect the relationship between the expression of IP-10 in serum, IP-10 mRNA in peripheral blood mononuclear cells (PBMCs), and the levels of HBV DNA in the serum of patients, and to explore their role in the pathogenesis of cirrhosis. METHODS: Typical patients with cirrhosis after HBV infection were selected, and their serum IP-10 concentrations were evaluated with ELISA, the content of IP-10 mRNA in PBMCs was measured by real-time PCR, and the load of HBV DNA in serum and PBMCs was assessed by semi-quantitative analysis of gel imaging. RESULTS: The levels of IP-10 in serum and IP-10 mRNA in PBMCs of patients with cirrhosis were 299.9 +/- 77.2 pg/ml and 0.7500 +/- 0.1495, respectively. They were higher than those of controls (P<0.05) and also increased in the HBV DNA(+) groups (P<0.05, P<0.01) to 343.0 +/- 80.3 pg/ml and 0.8465 +/- 0.1528, respectively. The levels of IP-10 in serum and IP-10 mRNA in PBMCs were clearly correlated with the load of HBV DNA (P<0.01). CONCLUSIONS: The levels of IP-10 and IP-10 mRNA in the peripheral blood of patients with cirrhosis increase are closely correlated with the load of HBV DNA in serum, and play a key role in the progression of post-hepatitic cirrhosis. (Hepatobiliary Pancreat Dis Int 2010; 9: 280-286)
基金Supported by grants-in-aid from the Key Project of Medical Science, No. RC2003100 and grants-in-aid from the project of Department of Health, No. H200523, Jiangsu Province, China
文摘AIM： To investigate the dynamic alteration of telomerase expression during development of hepatocellular carcinoma （HCC） and its diagnostic implications in liver tissues or peripheral blood mononuclear cells for HCC. METHODS： Dynamic expressions of liver telomerase during malignant transformation of hepatocytes were observed in Sprague-Dawly （SD） rats fed with 0.05% of 2-fluoenyacetamide （2-FAA）. Total RNA and telomerase were extracted from rat or human liver tissues. The telomerase activities in livers and in circulating blood were detected by a telomeric repeat amplification protocol-enzyme-linked immunosorbent assay （TRAP- ELISA）, and its diagnostic value was investigated in patients with benign or malignant liver diseases. RESULTS： The hepatoma model displayed the dynamic expression of hepatic telomerase during HCC development. The telomerase activities were consistent with liver total RNA levels （r = 0.83, P 〈 0.01） at the stages of degeneration, precancerosis, and cancerization of hepatocytes. In HCC patients, the telomerase levels in HCC tissues were significantly higher than in their adjacent non-cancerous tissues, but liver total RNA levels were lower in the former than in the latter. Although the circulating telomerase of HCC patients was abnormally expressed among patients with chronic liver diseases, the telomerase activity was a non-specific marker for HCC diagnosis, because the incidence was 15.7% in normal control, 25% in chronic hepatitis, 45.9% in liver cirrhosis, and 85.2% in HCC, respectively when absorbance value of telomerase activity was more than 0.2. If the value was over 0.6, the incidence was 60% in HCC group and 0% in any of the others （P 〈 0.01） except in two cases with liver cirrhosis. However, the combination of circulating telomerase with serum alpha-fetoprotein level could increase the positive rate and the accuracy （92.6%, 125 of 135） of HCC diagnosis. CONCLUSION： The overexpression of telomerase is associated with HCC development, and its abnormality in liver tissues or in peripheral blood could be a useful marker for diagnosis and prognosis of HCC.
基金This project was supported by a program of Science Project of Hubei Province (No.2003AA301C10).
文摘The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.
基金Supported by the Innovation Foundation of Wuhan University,No.301270054
文摘AIM： To study the dynamic changes of hepatits B virus （HBV） DNA in serum and peripheral blood mononuclear cells （PBMCs） of patients after lamivudine therapy. METHODS： A total of 72 patients with chronic HBV infection were included in this study. All patients were confirmed to have the following conditions： above 16 years of age, elevated serum alanine amonotransferase （ALT）, positive hepatitis B e antigen （HBeAg）, positive HBV DNA in serum and PBMCs, negative antibodies against HAV, HCV, HDV, HEV. Other possible causes of chronic liver damages, such as drugs, alcohol and autoimmune diseases were excluded. Seventy-two cases were randomly divided into lamivudine treatment group （n = 42） and control group （n = 30）. HBV DNA was detected both in serum and in PBMCs by fluorescence quantitative polymerase chain reaction （PCR）, during and after lamivudine treatment. RESULTS： In the treatment group, HBV DNA became negative both in serum and in PBMC, of, 38 and 25 out of 42 cases respectively during the 48 wk oflamivudine treatment, the negative rate was 90.5% and 59.5% respectively. In the control group, the negative rate was 23.3% and 16.7% respectively. It was statistically significant at 12, 24 and 48 wk as compared with the control group （P 〈 0.005）. The average conversion period of HBV DNA was 6 wk （2-8 wk） in serum and 16 wk （8-24 wk） in PBMC.CONCLUSION： Lamivudine has remarkable effects on HBV replication both in serum and The inhibitory effect on HBV DNA in PBMCs than that in serum inhibitory in PBMCs. is weaker
基金supported by the 2019 Yeungnam University Research Grant。
文摘Astragalus membranaceus(A.membranaceus)is a widely used traditional herb in China and Korea.A.membranaceus polysaccharides(AMP),which make up a major part of the root extract,have been shown to modulate immune modulations,especially activation of bone marrow-derived dendritic cells(BMDCs)and T cells.However,the immune stimulatory effect of AMP in the mouse in vivo and human peripheral blood DCs(PBDCs)has not been well investigated.In this study,we found that intravenous(i.v.)injection of AMP in C57 BL/6 mice induced remarkable elevations in co-stimulatory and MHC class I and II molecule levels in the splenic DCs and its subsets.The stimulatory effect of DCs by AMP was elevated 6 h after treatment,which rapidly decreased 18 h after injection.Furthermore,AMP promoted intracellular production of pro-inflammatory cytokines in spleen DC subsets,which contributed elevation of serum cytokine levels.Finally,the AMP promoted PBDC activation.Thus,these results demonstrate that AMP can be used as an immune stimulatory molecules in human and mouse.
基金supported by a grant from Natural Sciences Foundation of Hubei Province, China (No. 2006ABA139)
文摘The aim of the present study was to investigate the expression of toll-like receptors （TLR） 9 in peripheral blood mononuclear cells （PBMC） of patients with chronic hepatitis B and C with different virus copies. The study group included 90 patients （60 with chronic hepatitis B, and 30 with chronic hepatitis C）, and 20 healthy people served as control group. The protein and mRNA levels of TLR9 were detected by using flow cytometry and real-time PCR. The serum viral copies of HBV and HCV were measured in all patients, and the correlation between HBV-DNA copies or HCV-RNA copies and the TLR9 expression was analyzed. Our results demonstrated that HBV or HCV infection led to a decreased expression of TLR9 mRNA and protein compared to the control group （P〈0.05）. The TLR9 protein and mRNA levels were negatively correlated with serum viral copies of HBV and HCV （r=-0.632, r=-0.909, P〈0.01）. It was concluded that TLR9 mRNA and protein are down-regulated in PBMC of HBV-infected or HCV-infected patients, and they are negatively correlated with serum viral copies and play an important role in detecting viral replication of HBV and HCV.
