Antifungal resistance is the leading cause of antifungal treatment failure in invasive candidiasis.Metabolic rewiring could become a new insight to account for antifungal resistance as to find innovative clinical ther...Antifungal resistance is the leading cause of antifungal treatment failure in invasive candidiasis.Metabolic rewiring could become a new insight to account for antifungal resistance as to find innovative clinical therapies.Here,we show that dynamic surface-enhanced Raman spectroscopy is a promising tool to identify the metabolic differences between fluconazole(Diflucan)-resistant and fluconazole(Diflucan)-sensitive Candida albicans through the signatures of biochemical components and complemented with machine learning algorithms and two-dimensional correlation spectroscopy,an underlying resistance mechanism,that is,the change of purine metabolites induced the resistance of Candida albicans has been clarified yet never reported anywhere.We hope the integrated methodology introduced in this work could be beneficial for the interpretation of cellular regulation,propelling the development of targeted antifungal therapies and diagnostic tools for more efficient management of severe antifungal resistance.展开更多
The goal of this study was to investigate the clinical application of free/total prostate-specific antigen(F/T PSA)ratio,considering the new broad serum total PSA(T-PSA)“gray zone”of 2.0–25.0 ng ml^(−1)in different...The goal of this study was to investigate the clinical application of free/total prostate-specific antigen(F/T PSA)ratio,considering the new broad serum total PSA(T-PSA)“gray zone”of 2.0–25.0 ng ml^(−1)in differential diagnosis of prostate cancer(PCa)and benign prostate diseases(BPD)in men over 50 years in Western China.A total of 1655 patients were included,528 with PCa and 1127 with BPD.Serum T-PSA,free PSA(F-PSA),and F/T PSA ratio were analyzed.Receiver operating characteristic curves were used to assess the efficiency of PSA and F/T PSA ratio.There were 47.4%of cancer patients with T-PSA of 2.0–25.0 ng ml^(−1).When T-PSA was 2.0–4.0 ng ml^(−1),4.0–10.0 ng ml^(−1),and 10.0–25.0 ng ml^(−1),the area under the curve(AUC)of F/T PSA ratio was 0.749,0.769,and 0.761,respectively.The best AUC of F/T PSA ratio was 0.811 when T-PSA was 2.0–25.0 ng ml^(−1),with a specificity of 0.732,a sensitivity of 0.788,and an optimal cutoff value of 15.5%.The AUC of F/T PSA ratio in different age groups(50–59 years,60–69 years,70–79 years,and≥80 years)was 0.767,0.806,0.815,and 0.833,respectively,and the best sensitivity(0.857)and specificity(0.802)were observed in patients over 80 years.The T-PSA trend was in accordance with the Gleason score,tumor node metastasis(TNM)stage,and American Joint Committee on Cancer prognosis group.Therefore,the F/T PSA ratio can facilitate the differential diagnosis of PCa and BPD in the broad T-PSA“gray zone”.Serum T-PSA can be a Gleason score and prognostic indicator.展开更多
Objective:A novel technique was explored using an airbag-selective portal vein blood arrester that circumvents the need for an intraoperative assessment of anatomical variations in patients with complex intrahepatic s...Objective:A novel technique was explored using an airbag-selective portal vein blood arrester that circumvents the need for an intraoperative assessment of anatomical variations in patients with complex intrahepatic space-occupying lesions.Methods:Rabbits undergoing hepatectomy were randomly assigned to 4 groups:intermittent portal triad clamping(PTC),intermittent portal vein clamping(PVC),intermittent portal vein blocker with an airbag-selective portal vein blood arrester(APC),and without portal blood occlusion(control).Hepatic ischemia and reperfusion injury were assessed by measuring the 7-day survival rate,blood loss,liver function,hepatic pathology,hepatic inflammatory cytokine infiltration,hepatic malondialdehyde levels,and proliferating cell nuclear antigen levels.Results:Liver damage was substantially reduced in the APC and PVC groups.The APC animals exhibited transaminase levels similar to or less oxidative stress damage and inflammatory hepatocellular injury compared to those exhibited by the PVC animals.Bleeding was significantly higher in the control group than in the other groups.The APC group had less bleeding than the PVC group because of the avoidance of portal vein skeletonization during hepatectomy.Thus,more operative time was saved in the APC group than in the PVC group.Moreover,the total 7-day survival rate in the APC group was higher than that in the PTC group.Conclusion:Airbag-selective portal vein blood arresters may help protect against hepatic ischemia and reperfusion injury in rabbits undergoing partial hepatectomy.This technique may also help prevent liver damage in patients requiring hepatectomy.展开更多
Introduction:Among all malignant tumors of the digestive system,pancreatic carcinoma exhibits the highest mortality rate.Currently,prevention and effective treatment are urgent issues that need to be addressed.Methods...Introduction:Among all malignant tumors of the digestive system,pancreatic carcinoma exhibits the highest mortality rate.Currently,prevention and effective treatment are urgent issues that need to be addressed.Methods:The study focused on meiotic nuclear divisions 1(MND1),integrating data from the Gene Expression Profiling Interactive Analysis(GEPIA)database with prognostic survival analysis.Simultaneously,experiments at cellular level were employed to demonstrate the effect of MND1 on the proliferation and migration of PC.The small-molecule inhibitor of MND1 was used to suppress the migration of PC cells by knocking down MND1 using small interfering RNA(siRNA)in Patu-8988 and Panc1 cell lines.Results:The results of Cell Counting Kit-8 indicated that the suppression of MND1 resulted in a decrease in cell proliferation.Wound healing and Transwell assays revealed that MND1 knockdown reduced cell migration and invasion.Flow cytometry revealed that inhibiting MND1 hindered the cell cycle.Furthermore,MND1 could stimulate the proliferation,migration,and invasion of Patu-8988 and Panc1 cells by increasing the expression of MND1.Notably,MND1 had a positive effect on H2AFX expression in PC cells.Elevated MND1 expression suggests the low overall survival rate of individuals diagnosed with PC.Conclusion:These findings suggest that MND1 has the potential to be a gene with the ability to accurately diagnose and treat PC.展开更多
Objective： The prevalence of non-alcoholic fatty liver disease （NAFLD） has markedly increased. Insulin resistance has been implicated in the pathogenesis of NAFLD. This study was aimed at observing the relationship...Objective： The prevalence of non-alcoholic fatty liver disease （NAFLD） has markedly increased. Insulin resistance has been implicated in the pathogenesis of NAFLD. This study was aimed at observing the relationship between insulin resistance and NAFLD, and evaluating the role of pioglitazone （PGZ） acting as insulin-sensitizing agents in the prevention and treatment of rat fatty liver induced by high fat feeding. Methods： The rats were separated randomly into 6 groups： model group Ⅰ were fed high fat diet for 8 weeks, PGZ prevention group were given PGZ 4 mg/（kg.d） simultaneously, while control group Ⅰ were fed normal food for 8 weeks; model group Ⅱ were fed high fat diet for 16 weeks, PGZ treatment group were given PGZ 4 mg/（kg.d） orally simultaneous with high fat diet for 8 weeks after high fat feeding for 8 weeks, control group Ⅱ were fed normal food for 16 weeks. The rats were sacrificed after 8 weeks and 16 weeks respectively. Liver weight, body weight, serum activities of alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, alkaline phosphatase （ALP）, tumor necrosis factor alpha （TNF-α）, fasting blood glucose （FBG）, fasting plasma insulin （FINS）, HOMA （homeostasis model assessment） insulin resistance index （HOMA-IR）, and the liver histology of rats of all groups were assayed. Results： After 8 weeks, the liver in model group Ⅰ showed typical steatosis, accompanied with mild to moderate lobular inflammatory cell infiltration, liver indexes and serum levels of ALT, AST, ALP, TNF-α were significantly increased （P〈0.05） compared with control group Ⅰ. Whereas, the degree of hepatic injury was attenuated in PGZ prevention group, liver indexes and serum levels of ALT, ALP were significantly decreased （P〈0.05） compared with model group Ⅰ. After 16 weeks, notable steatosis, and lobular inflammation were observed in model group Ⅱ rat liver, while the degree of hepatic injury was attenuated in the PGZ treatment group. Liver index, serum levels ofALT, AST, ALP, FINS and HOMA-IR were significantly increased （P〈0.05） in model group Ⅱ compared with control group Ⅱ. Whereas, in PGZ treatment group, serum levels of AST and FINS showed decreasing tendency, liver indexes, serum levels of ALT, ALP, TNF-α and HOMA-IR were significantly decreased compared with model group Ⅱ. Conclusion： Insulin resistance plays a role in the pathogenesis of NAFLD in rats. Pioglitazone can attenuate insulin resistance and biochemical and histological injury in high fat-induced fatty liver in rats.展开更多
Objective： Previous studies have investigated the role of cytotoxic T-lymphocyte antigen-4 （CTLA-4） and tumor necrosis factor-alpha （TNF-a） in carcinogenesis of osteosarcoma, but their results were inconsistent. ...Objective： Previous studies have investigated the role of cytotoxic T-lymphocyte antigen-4 （CTLA-4） and tumor necrosis factor-alpha （TNF-a） in carcinogenesis of osteosarcoma, but their results were inconsistent. We aimed to clarify the associations between CTLA-4, TNF-a polymorphism and osteosarcoma risk by using meta-analysis. Methods： We searched relevant studies without language restriction in PubMed, EMbase, Cochrane Library, Google Scholar databases, Chinese National Knowledge Infrastructure （CNKI） and conference literature in humans published prior to March 2013. The strengths of the associations between genetic variants and osteosarcoma risk were estimated by odds ratio （OR） with 95% confidence interval （95% CI）. Results： A total of seven studies with 1,198 osteosarcoma patients and 1,493 controls were selected. Four studies were eligible for CTLA-4 （1,003 osteosarcoma and 1,162 controls）, and three studies for TNF-a （195 osteosarcoma and 331 controls）. Pooled results showed that rs231775 polymorphism of CTLA-4 was associated with osteosarcoma risk （GG vs. AA： OR=1.63, 95% CI=1.24-2.13; GG ＋ GA vs. AA： OR=1.56, 95% CI=1.21-2.01; AA ＋ GA vs. GG： OR=0.83, 95% CI=0.71-0.97; G vs. A： OR=1.21, 95% CI=1.08-1.36）. No significant heterogeneity was observed across the studies. No significant associations were found between rs5742909 polymorphism of CTLA-4 or rs1800629 polymorphism of TNF-a and osteosarcoma risk. Conclusions： These results suggest that the rs231775 polymorphism of CTLA-4 may play an important role in carcinogenesis of osteosarcoma.展开更多
Ticks are considered the second most common pathogen vectors transmitting a broad range of vital human and veterinary viruses.From 2017 to 2018,640 ticks were collected in eight different provinces in central and west...Ticks are considered the second most common pathogen vectors transmitting a broad range of vital human and veterinary viruses.From 2017 to 2018,640 ticks were collected in eight different provinces in central and western China.Six species were detected,including H.longicornis,De.everestianus,Rh.microplus,Rh.turanicus,Rh.sanguineous,and Hy.asiaticum.Sixty-four viral metagenomic libraries were constructed on the MiSeq Illumina platform,resulting in 13.44 G(5.88×10^(7))of 250-bp-end reads,in which 2,437,941 are viral reads.We found 27 nearly complete genome sequences,including 16 genome sequences encoding entire protein-coding regions(lack of 30 or 50 end non-coding regions)and complete viral genomes,distributed in the arboviral family(Chuviridae,Rhabdoviridae,Nairoviridae,Phenuiviridae,Flaviviridae,Iflaviridae)as well as Parvoviridae and Polyomaviridae that cause disease in mammals and even humans.In addition,13 virus sequences found in Chuviridae,Nairoviridae,Flaviviridae,Iflaviridae,Hepeviridae,Parvoviridae,and Polyomaviridae were identified as belonging to a new virus species in the identified viral genera.Besides,an epidemiological survey shows a high prevalence(9.38%and 15.63%)of two viruses(Ovine Copiparvovirus and Bovine parvovirus 2)in the tick cohort.展开更多
AIM To investigate and compare the analytical and clinical performance of Tian Long automatic hypersensitive hepatitis B virus(HBV) DNA quantification system and Roche CAP/CTM system.METHODS Two hundred blood samples ...AIM To investigate and compare the analytical and clinical performance of Tian Long automatic hypersensitive hepatitis B virus(HBV) DNA quantification system and Roche CAP/CTM system.METHODS Two hundred blood samples for HBV DNA testing, HBV-DNA negative samples and high-titer HBV-DNA mixture samples were collected and prepared. National standard materials for serum HBV and a worldwide HBV DNA panel were employed for performance verification. The analytical performance, such as limit of detection, limit of quantification, accuracy, precision, reproducibility, linearity, genotype coverage and cross-contamination, was determined using the Tian Long automatic hypersensitive HBV DNA quantification system(TL system). Correlation and Bland-Altman plot analyses were carried out to compare the clinical performance of the TL system assay and the CAP/CTM system. RESULTS The detection limit of the TL system was 10 IU/m L, and its limit of quantification was 30 IU/m L. The differences between the expected and tested concentrations of the national standards were less than ± 0.4 Log10 IU/m L, which showed high accuracy of the system. Results of the precision, reproducibility and linearity tests showed that the multiple test coefficient of variation(CV) of the same sample was less than 5% for 102-106 IU/m L; and for 30-108 IU/m L, the linear correlation coefficient r2 = 0.99. The TL system detected HBV DNA(A-H) genotypes and there was no cross-contamination during the "checkerboard" test. When compared with the CAP/CTM assay, the two assays showed 100% consistency in both negative and positive sample results(15 negative samples and 185 positive samples). No statistical differences between the two assays in the HBV DNA quantification values were observed(P > 0.05). Correlation analysis indicated a significant correlation between the two assays, r2 = 0.9774. The Bland-Altman plot analysis showed that 98.9% of the positive data were within the 95% acceptable range, and the maximum difference was-0.49.CONCLUSION The TL system has good analytical performance, and exhibits good agreement with the CAP/CTM system in clinical performance.展开更多
Little data is available on the evaluation of the occurrence rates of Epstein-Barr virus(EBV) in saliva and relationship with highly active antiretroviral therapy(HAART) use in HIV/AIDS patients in China. We conducted...Little data is available on the evaluation of the occurrence rates of Epstein-Barr virus(EBV) in saliva and relationship with highly active antiretroviral therapy(HAART) use in HIV/AIDS patients in China. We conducted a retrospective cohort study of EBV serological tests for HIV/AIDS patients who were treated in the hospitals for infectious diseases in Wuxi and Shanghai, China from May 2016 to April 2017. The EBV-seropositive samples were identified by ELISA. EBV-specific primers and probes were used for the quantitative detection of viral DNA from saliva via quantitative real-time polymerase chain reaction. CD4 cell counts of the HIV/AIDS patients were detected by a flow cytometry. A total of 372 HIV/AIDS patients were ultimately selected and categorized for this retrospective cohort study. For EBV IgG and IgM, the HIV/AIDS HAART use(H) and non-HAART use(NH) groups had significantly higher seropositive rates than the HIV-negative control group. The HIV/AIDS(NH) group had the highest seropositive rate(IgG, 94.27%; IgM, 68.98%) and the highest incidence of EBV reactivation or infection. For salivary EBV DNA-positive rates and quantities, the HIV/AIDS(H)(73.69%) and the HIV/AIDS(NH)(100%) groups showed significantly higher values than the HIV-negative control group(35.79%,[ twofold). Further, the salivary EBV DNA-negative population had significantly higher CD4 cell counts than the EBV DNA-positive population in the HIV/AIDS(H) group and the HIV/AIDS(NH) groups. Thus, HAART use is beneficial in decreasing the EBV salivary shedding in HIV/AIDS patients and indirectly decreases EBV transmission risk.展开更多
Columbianetin acetate(CE)is one of the effective components of Angelica pubescens.So far,the specific role and molecular mechanism of CE in pancreatic cancer are not clear.Thus,this study aimed to explore the specific...Columbianetin acetate(CE)is one of the effective components of Angelica pubescens.So far,the specific role and molecular mechanism of CE in pancreatic cancer are not clear.Thus,this study aimed to explore the specific mechanism of CE on pancreatic cancer.The target genes combined with CE were predicted through the PharmMapper database and the 3D molecular structure of CE.Then,the Cancer Genome Atlas(TCGA)and Cistrome data browser(DB)databases were used to screen Meiotic nuclear divisions 1(MND1)-related genes,transcription factors,and transcription factor data sets,and the intersection of the above data sets.The“limma”package in the R and gene expression profiling interactive analysis(GEPIA)databases were used to analyze the correlation and survival difference between the target genes and MND1 to predict the degree of association between CE and MND1.Western blotting and RT-PCR experiments revealed the regulatory relationship among CE,E2F1,and MND1 at the cellular level.The specific effects of CE on pancreatic cancer cells were explored through CCK8,wound healing,migration,and flow cycle experiments.E2F1,also the predictive transcription factor of MND1,was also the predictive target protein of CE.At the same time,E2F1 and MND1 were closely related in pancreatic tissue.In the cell function experiment,CE and interference with E2F1 expression could reduce the gene and protein expression of MND1,which was closely associated with cell proliferation,migration,and cycle development.Similarly,interfering with the expression of mnd1 can also inhibit the further development of tumor cells.CE may inhibit the development of pancreatic cancer cells by reducing the expression of MND1.This implies that CE may be a potential novel agent for the treatment of pancreatic cancer.展开更多
To establish a rapid identification method for common pathogenic bacteria on the basis of molecular biology and to construct a preliminary Polymerase Chain Reaction-Capillary Electrophoresis - Restriction Fragment Len...To establish a rapid identification method for common pathogenic bacteria on the basis of molecular biology and to construct a preliminary Polymerase Chain Reaction-Capillary Electrophoresis - Restriction Fragment Length Polymorphism (PCR-CE-RFLP) database of bacteria isolated from clinical specimens frequently, 183 strains collected from clinical samples belonging to 12 genera and 19 species whose biochemical characterizations corresponded to the typical ones were examined. The genomic DNAs were amplified by two pairs of fluorescence labeled primers aiming at 16S rRNA gene and 16S-23S rRNA spacer region gene respectively at the same time. PCR products were then digested by restriction endonuclease HaeⅢ incompletely before taking capillary electrophoresis. The results with the PCR-CE-RFLP patterns of 16S rRNA genes were just alike within some genera, but when it comes to 16S-23S rRNA spacer region genes, each bacterium showed a unique pattern, which can be distinguished from each other easily. It seems that PCR-CE-RFLP patterns of 16S rRNA gene could only be used to classify the bacteria into family level, whereas the data of 16S-23S rRNA spacer region gene could be utilized to identify the whole microorganisms as precisely as the species level. In spite of the data of the spacer region gene alone can be sufficiently to verify the whole bacteria, we insist that the 16S rRNA gene could be of some assistant in case that there should be lots of families of bacteria, in which some similar ones, with the same RFLP data of 16S-23S rRNA spacer region gene, may coexist. This study proves that the utility of PCR-CE-RFLP is a convenient, rapid method to identify pathogenic bacteria, and is also a quick diagnosis measure for application to clinical use.展开更多
Developing accurate and sensitive DNA methyltransferase(MTase) analysis methods is essential for early clinical diagnosis and development of antimicrobial drug targets. In this work, by coupling WO_(3-x) dotsencapsula...Developing accurate and sensitive DNA methyltransferase(MTase) analysis methods is essential for early clinical diagnosis and development of antimicrobial drug targets. In this work, by coupling WO_(3-x) dotsencapsulated metal-organic frameworks(MOFs) as co-reactants and terminal deoxynucleotidyl transferase(Td T)-mediated template-free branched polymerization, a dual signal-amplified electrochemiluminescent(ECL) biosensor was constructed to detect DNA adenine methylation(Dam) MTase. The employment of WO_(3-x) dots-encapsulated MOFs(i.e., NH_(2)-UIO66@WO_(3-x) ) was not only beneficial for biomolecule conjugation because of the abundant amino groups but also led to a 7-fold enhanced ECL response due to the increased loading of WO_(3-x). Moreover, Td T-mediated template-free branched polymerization promoted the capture of ECL emitters on the electrode surface, achieving 20-fold enhanced signal amplification. The presented ECL biosensor demonstrated a low detection limit of 2.4 × 10^(-4)U/m L, and displayed high reliability for the detection of Dam MTase in both spiked human serum and E. coli cell samples, and for the screening of potential inhibitors. This study opens a new avenue for designing a dual signal amplificationbased ECL bioassay for Dam MTase and screening inhibitors in the fields of clinical diagnosis and drug development.展开更多
Spermatogenesis, maturation, capacitation and fertilization are precisely regulated by glycosylation. However, the relationship between altered glycosylation patterns and the onset and development of reproductive diso...Spermatogenesis, maturation, capacitation and fertilization are precisely regulated by glycosylation. However, the relationship between altered glycosylation patterns and the onset and development of reproductive disorders is unclear, mainly limited by the lack of in situ imaging techniques for spermatozoa glycosylation. We developed an efficient and highly specific spermatozoa glycan imaging technique based on the robust chemoselective labeling of sialic acid(Sia) and N-acetyl-D-galactosamine(Gal/GalNAc). We further proposed a “tandem glycan chemoselective labeling” strategy to achieve simultaneous imaging of two types of glycans on spermatozoa. We applied the developed method to the spermatozoa from oligozoospermic patients and diabetic mice and found that these spermatozoa showed higher levels of Sia and Gal/Gal NAc expression than the normal groups. Moreover, spermatozoa from diabetic mice showed a severe decrease in number, viability, and forward motility, suggesting that in vivo glucose metabolism disorders may lead to an elevated level of spermatozoa glycosylation and have a correlation with the development of oligoasthenotspermia. Our work provides a research tool to reveal the relationship between glycosylation modification and spermatozoa quality, and a promising clue for the development of glycan-based reproductive markers.展开更多
Posttranscriptional RNA modification is an important mode of epigenetic regulation in various biological and pathological contexts.N6,2′-O-dimethyladenosine(m6Am)is one of the most abundant methylation modifications ...Posttranscriptional RNA modification is an important mode of epigenetic regulation in various biological and pathological contexts.N6,2′-O-dimethyladenosine(m6Am)is one of the most abundant methylation modifications in mammals and usually occurs at the first transcribed nucleotide.Accumulating evidence indicates that m6Am modifications have important roles in RNA metabolism and physiological and pathological processes.PCIF1(phosphorylated C-terminal domain interacting factor 1)is a protein that can bind to the phosphorylated C-terminal domain of RNA polymeraseⅡthrough its WW domain.PCIF1 is named after this binding ability.Recently,PCIF1 has been identified as a cap-specific adenine N6-methyltransferase responsible for m6Am formation.Discovered as the sole m6Am methyltransferase for mammalian mRNA,PCIF1 has since received more extensive and in-depth study.Dysregulation of PCIF1 contributes to various pathological processes.Targeting PCIF1 may hold promising therapeutic significance.In this review,we provide an overview of the current knowledge of PCIF1.We explore the current understanding of the structure and the biological characteristics of PCIF1.We further review the molecular mechanisms of PCIF1 in cancer and viral infection and discuss its therapeutic potential.展开更多
Metastasis-associated lung adenocarcinoma transcript 1(MALAT1)is a well-established oncogenic long non-coding RNA,the higher expression of which is strongly correlated with cancer events such as tumorigenesis,progress...Metastasis-associated lung adenocarcinoma transcript 1(MALAT1)is a well-established oncogenic long non-coding RNA,the higher expression of which is strongly correlated with cancer events such as tumorigenesis,progression,metastasis,drug resistance,and treatment outcome in solid cancers.Recently,a series of studies has highlighted its potential role in hematological malignancies in terms of these events.Similar to solid cancers,MALAT1 can regulate various target genes via sponging and epigenetic mechanisms,but the miRNAs sponged by MALAT1 differ from those identified in solid cancers.In this review,we systematically describe the role and underlying mechanisms of MALAT1 in multiple types of hematological malignancies,including regulation of cell proliferation,metastasis,stress response,and glycolysis.Clinically,MALAT1 expression is related to poor treatment outcome and drug resistance,therefore exhibiting potential prognostic value in multiple myeloma,lymphoma,and leukemia.Finally,we discuss the evaluation of MALAT1 as a novel therapeutic target against cancer in preclinical studies.展开更多
To the Editor:Current combined antiretroviral therapy(cART)can effectively suppress the plasma viral load in most people living with human immunodeficiency virus-1(HIV,PLWH).However,approximately 20-30%of PLWH undergo...To the Editor:Current combined antiretroviral therapy(cART)can effectively suppress the plasma viral load in most people living with human immunodeficiency virus-1(HIV,PLWH).However,approximately 20-30%of PLWH undergoing cART can have very low but detectable plasma viral loads(<20 copies/mL,the limit of quantification in commercially available assays),known as residual HIV-1 low-level viremia(LLV).^([1])Residual HIV-1 LLV detection can reflect the source and dynamics of the HIV-1 reservoir in PLWH,thus playing an important role in both HIV-1 management and basic research.^([2])展开更多
Background:Autophagy death of cancer cells is detrimental to apoptosis induced by therapeutic drugs,which promotes tumor progression to a certain extent.Increasing reports have demonstrated the regulatory role of circ...Background:Autophagy death of cancer cells is detrimental to apoptosis induced by therapeutic drugs,which promotes tumor progression to a certain extent.Increasing reports have demonstrated the regulatory role of circular RNAs(circRNAs)in autophagy.Here,we aimed to determine the role of hsa_circ_0009109 in autophagy in gastric cancer(GC).Methods:The effects of hsa_circ_0009109 on autophagy were examined using quantitative real-time polymerase chain reaction(qPCR),transmission electron microscopy,Western blot,and immunofluorescence.The mechanism of hsa_circ_0009109 regulating the miR-544a-3p/bcl-2 axis was analysed using fluorescence in situ hybridization,dual-luciferase reporter,and rescue experiments.Results:Functional testing indicated that hsa_circ_0009109 was significantly down-expressed in GC tissues and cell lines.A reduction in cytoplasmic-derived hsa_circ_0009109 could promote GC progression by accelerating cell proliferation,enhancing migration and invasion,inhibiting apoptosis,and accelerating the cell cycle progression.Besides,hsa_circ_0009109 was found to exert the effect of an autophagy inhibitor such as 3-Methyladenine(3-MA),which was manifested by the weakening of the immunofluorescence of LC3B and the reduction in autophagy-related proteins after overexpression of hsa_circ_0009109,while increased autophagosomes were observed after interference with hsa_circ_0009109.Subsequently,the crosstalk between hsa_circ_0009109 and miR-544a-3p/bcl-2 was verified using dual-luciferase reporter assay.The autophagy status was altered under the regulation of the hsa_circ_0009109-targeted miR-544a-3p/bcl-2 axis.Conclusions:The hsa_circ_0009109 mediated a novel autophagy regulatory network through targeting the miR-544a-3p/bcl-2 axis,which may shed new light on the exploration of therapeutic targets for the clinical treatment of GC.展开更多
Background:Metastasis is the main cause of death in colorectal cancer(CRC).Metastasis is a sequential and dynamic process,but the development of tumor cells during this process is unclear.In this study,we aimed to rev...Background:Metastasis is the main cause of death in colorectal cancer(CRC).Metastasis is a sequential and dynamic process,but the development of tumor cells during this process is unclear.In this study,we aimed to reveal characteristics of tumor cell subset during CRC metastasis.Methods:Single-cell RNA sequence CRC data of normal epithelium,non-metastatic primary tumor,metastatic primary tumor,and liver metastases from gene expression omnibus(GEO)dataset were analyzed to reveal characteristics of CRC metastasis.Primary tu-mor tissues of three non-metastatic CRC and three metastatic CRC patients from Union Hospital of Tongji Medical College(Wuhan,China)were used to verify the characteristics of CRC metastasis.Results:We identified a metastasis-related cancer cell subset EP1,which was characterized with a high expression of KRT17,LAMC2,EMP1,and PLAC8.EP1 had an enhanced cell-cell interaction,which interacted with SPPþmacrophages and drove them toward anti-inflammatory and immunosuppressive phenotype.Dynamic changes in genes and TF regulons during the metastasis were also revealed.Conclusions:This study advanced our understanding of the development of tumor cells during CRC metastasis and further identi-fied metastasis-related subset and potential therapeutic targets for the treatment and prevention of CRC metastasis.展开更多
Hypoxia,a prevalent characteristic of solid tumors,substantially impairs the efficacy of cancer treatments.However,there are no feasible clinical approaches for treating hypoxic tumors.Here,we develop metal-phenolic n...Hypoxia,a prevalent characteristic of solid tumors,substantially impairs the efficacy of cancer treatments.However,there are no feasible clinical approaches for treating hypoxic tumors.Here,we develop metal-phenolic networks(CuGI)utilizing the natural glycolysis inhibitor(epigallocatechin gallate)and the essential metal element in the human body(copper ions),specifically targeting and annihilating hypoxic cancer cells.CuGI redirects the metabolic pathway of hypoxic cancer cells from anaerobic glycolysis to oxidative phosphorylation,thereby enhancing reactive oxygen species production and promoting oligomerization of lipoylated proteins in the tricarboxylic acid cycle.Through targeted induction of oxidative and proteotoxic stresses,CuGI induces apoptosis and cuproptosis specifically in cancer cells under hypoxic conditions while sparing normal cells.Moreover,cancer cell membrane-coated CuGI(CuGI@CM)exhibits enhanced tumor penetration effect and demonstrates commendable biocompatibility,effectively suppressing colorectal tumor growth.Importantly,CuGI@CM,when combined with vascular disruptors or radiotherapy which aggravate tumor hypoxia,syner-gistically potentiates therapeutic efficacy.Thus,CuGI represents a specific and potent nanotherapeutic capable of selectively eliminating hypoxic tumors,offering promise in combination therapies to address tumor hypoxia.展开更多
Background The normal microbial flora of the vagina plays an important role in preventing genital and urinary tract infections in women. Thus an accurate understanding of the composition and ecology of the ecosystem i...Background The normal microbial flora of the vagina plays an important role in preventing genital and urinary tract infections in women. Thus an accurate understanding of the composition and ecology of the ecosystem is important to understanding the etiology of these diseases. This study aimed to compare the characteristics of main lactobacillus species between healthy women and women with bacterial vaginosis （BV） by quantitative culture and PCR methods. Main lactobacillus species include L. crispatus , L. gasseri, L. jensenii and L. iners.Methods A total of 150 Women attending Gynecology Outpatient Clinic of Beijing Friendship Hospital, were diagnosed as having BV because three or more of the following criteria were met （standard of Amsel＇s composite criteria）： homogeneous discharge, elevated vaginal pH （pH 〉4.5）, production of amines, and presence of ＂clue＂ cells. Those with less than three of the criteria were considered as healthy. Simultaneously, smears were made of vaginal fluid and Gram stained, then were assessed qualitatively as normal （grade Ⅰ）, intermediate （grade Ⅱ）, or consistent with BV （grade Ⅲ）. Gardnerella vaginalis were identified by using Vitek 2 Compact and PCR methods. Lactobacillus species were identified by PCR methods. Gardnerella vaginalis and lactobacilli colony counts were determined by calculating the most number of colonies of each species in the appropriate plates （colonies between 10 and 300）, corrected by the dilution of the sample in the plates, and multiplied by 10 （to account for plating 100 μl）, in order to get colony forming units per milliliter of vaginal secretion.Results BV was diagnosed in 31% （46/150） patients using the composite criteria, the remainder being regarded as healthy. The majority of patients with BV had a smear assessed as grade Ⅲ （91%, 42/46） and minority of them had a smear assessed as grade Ⅱ（9%, 4/46）. The majority of healthy women had a smear assessed as grade Ⅰ （64%, 67/104）. Smears assessed as grade Ⅱwere found （36%, 37/104） among patients diagnosed as healthy, with less than three of the composite criteria. L. crispatus was cultured from 94% of healthy women and 83% of women with BV, with the former colonies count average value of 106 and the latter of 103. L. gasseri, L. iners, and L. jensenfi were cultured from 85%, 68% and 43% of healthy women; and 28%, 89% and 44% of BV women, respectively.Conclusions The quantities of four lactobacillus species except L. jensenii had a significant difference between healthy women and women with BV. Our results provide support for the negative association between L. iners and L. gasseri. Although L. crispatus were existent both in healthy and BV positive women＇s vagina, the numbers of L. crispatus were significantly different for the dominant number in healthy women. Smears of vaginal fluid and Gram stain play an important guiding role in bacteria culture.展开更多
基金supported by grants from the National Natural Science Foundation of China(Nos.22074015 and 82074428)Youth Talent Cultivation Initiation Fund of Zhongda Hospital,Southeast University(No.CZXM-GSP-RC110)to Hao Li+1 种基金Evidence-Based Capacity Building for TCM Specialty Therapies for Skin Diseases of National Administration of TCMInnovative Team Projects of Shanghai Municipal Commission of Health(No.2022CX011)to Fulun Li.
文摘Antifungal resistance is the leading cause of antifungal treatment failure in invasive candidiasis.Metabolic rewiring could become a new insight to account for antifungal resistance as to find innovative clinical therapies.Here,we show that dynamic surface-enhanced Raman spectroscopy is a promising tool to identify the metabolic differences between fluconazole(Diflucan)-resistant and fluconazole(Diflucan)-sensitive Candida albicans through the signatures of biochemical components and complemented with machine learning algorithms and two-dimensional correlation spectroscopy,an underlying resistance mechanism,that is,the change of purine metabolites induced the resistance of Candida albicans has been clarified yet never reported anywhere.We hope the integrated methodology introduced in this work could be beneficial for the interpretation of cellular regulation,propelling the development of targeted antifungal therapies and diagnostic tools for more efficient management of severe antifungal resistance.
基金This study was supported by the Sichuan Science and Technology Program(No.2020YFS0137).
文摘The goal of this study was to investigate the clinical application of free/total prostate-specific antigen(F/T PSA)ratio,considering the new broad serum total PSA(T-PSA)“gray zone”of 2.0–25.0 ng ml^(−1)in differential diagnosis of prostate cancer(PCa)and benign prostate diseases(BPD)in men over 50 years in Western China.A total of 1655 patients were included,528 with PCa and 1127 with BPD.Serum T-PSA,free PSA(F-PSA),and F/T PSA ratio were analyzed.Receiver operating characteristic curves were used to assess the efficiency of PSA and F/T PSA ratio.There were 47.4%of cancer patients with T-PSA of 2.0–25.0 ng ml^(−1).When T-PSA was 2.0–4.0 ng ml^(−1),4.0–10.0 ng ml^(−1),and 10.0–25.0 ng ml^(−1),the area under the curve(AUC)of F/T PSA ratio was 0.749,0.769,and 0.761,respectively.The best AUC of F/T PSA ratio was 0.811 when T-PSA was 2.0–25.0 ng ml^(−1),with a specificity of 0.732,a sensitivity of 0.788,and an optimal cutoff value of 15.5%.The AUC of F/T PSA ratio in different age groups(50–59 years,60–69 years,70–79 years,and≥80 years)was 0.767,0.806,0.815,and 0.833,respectively,and the best sensitivity(0.857)and specificity(0.802)were observed in patients over 80 years.The T-PSA trend was in accordance with the Gleason score,tumor node metastasis(TNM)stage,and American Joint Committee on Cancer prognosis group.Therefore,the F/T PSA ratio can facilitate the differential diagnosis of PCa and BPD in the broad T-PSA“gray zone”.Serum T-PSA can be a Gleason score and prognostic indicator.
基金supported by the Hainan Provincial Natural Science Foundation of China(No.821QN0982 and No.2019RC373)。
文摘Objective:A novel technique was explored using an airbag-selective portal vein blood arrester that circumvents the need for an intraoperative assessment of anatomical variations in patients with complex intrahepatic space-occupying lesions.Methods:Rabbits undergoing hepatectomy were randomly assigned to 4 groups:intermittent portal triad clamping(PTC),intermittent portal vein clamping(PVC),intermittent portal vein blocker with an airbag-selective portal vein blood arrester(APC),and without portal blood occlusion(control).Hepatic ischemia and reperfusion injury were assessed by measuring the 7-day survival rate,blood loss,liver function,hepatic pathology,hepatic inflammatory cytokine infiltration,hepatic malondialdehyde levels,and proliferating cell nuclear antigen levels.Results:Liver damage was substantially reduced in the APC and PVC groups.The APC animals exhibited transaminase levels similar to or less oxidative stress damage and inflammatory hepatocellular injury compared to those exhibited by the PVC animals.Bleeding was significantly higher in the control group than in the other groups.The APC group had less bleeding than the PVC group because of the avoidance of portal vein skeletonization during hepatectomy.Thus,more operative time was saved in the APC group than in the PVC group.Moreover,the total 7-day survival rate in the APC group was higher than that in the PTC group.Conclusion:Airbag-selective portal vein blood arresters may help protect against hepatic ischemia and reperfusion injury in rabbits undergoing partial hepatectomy.This technique may also help prevent liver damage in patients requiring hepatectomy.
基金supported by grants from National Innovation Program for College Students(202210367076)Graduate Student Research Innovation Program of Bengbu Medical College(Byycxz22016)the National Natural Science Foundation of China(82072585),and the Key Research Project of Bengbu Medical College(No.2020byzd029).
文摘Introduction:Among all malignant tumors of the digestive system,pancreatic carcinoma exhibits the highest mortality rate.Currently,prevention and effective treatment are urgent issues that need to be addressed.Methods:The study focused on meiotic nuclear divisions 1(MND1),integrating data from the Gene Expression Profiling Interactive Analysis(GEPIA)database with prognostic survival analysis.Simultaneously,experiments at cellular level were employed to demonstrate the effect of MND1 on the proliferation and migration of PC.The small-molecule inhibitor of MND1 was used to suppress the migration of PC cells by knocking down MND1 using small interfering RNA(siRNA)in Patu-8988 and Panc1 cell lines.Results:The results of Cell Counting Kit-8 indicated that the suppression of MND1 resulted in a decrease in cell proliferation.Wound healing and Transwell assays revealed that MND1 knockdown reduced cell migration and invasion.Flow cytometry revealed that inhibiting MND1 hindered the cell cycle.Furthermore,MND1 could stimulate the proliferation,migration,and invasion of Patu-8988 and Panc1 cells by increasing the expression of MND1.Notably,MND1 had a positive effect on H2AFX expression in PC cells.Elevated MND1 expression suggests the low overall survival rate of individuals diagnosed with PC.Conclusion:These findings suggest that MND1 has the potential to be a gene with the ability to accurately diagnose and treat PC.
文摘Objective： The prevalence of non-alcoholic fatty liver disease （NAFLD） has markedly increased. Insulin resistance has been implicated in the pathogenesis of NAFLD. This study was aimed at observing the relationship between insulin resistance and NAFLD, and evaluating the role of pioglitazone （PGZ） acting as insulin-sensitizing agents in the prevention and treatment of rat fatty liver induced by high fat feeding. Methods： The rats were separated randomly into 6 groups： model group Ⅰ were fed high fat diet for 8 weeks, PGZ prevention group were given PGZ 4 mg/（kg.d） simultaneously, while control group Ⅰ were fed normal food for 8 weeks; model group Ⅱ were fed high fat diet for 16 weeks, PGZ treatment group were given PGZ 4 mg/（kg.d） orally simultaneous with high fat diet for 8 weeks after high fat feeding for 8 weeks, control group Ⅱ were fed normal food for 16 weeks. The rats were sacrificed after 8 weeks and 16 weeks respectively. Liver weight, body weight, serum activities of alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, alkaline phosphatase （ALP）, tumor necrosis factor alpha （TNF-α）, fasting blood glucose （FBG）, fasting plasma insulin （FINS）, HOMA （homeostasis model assessment） insulin resistance index （HOMA-IR）, and the liver histology of rats of all groups were assayed. Results： After 8 weeks, the liver in model group Ⅰ showed typical steatosis, accompanied with mild to moderate lobular inflammatory cell infiltration, liver indexes and serum levels of ALT, AST, ALP, TNF-α were significantly increased （P〈0.05） compared with control group Ⅰ. Whereas, the degree of hepatic injury was attenuated in PGZ prevention group, liver indexes and serum levels of ALT, ALP were significantly decreased （P〈0.05） compared with model group Ⅰ. After 16 weeks, notable steatosis, and lobular inflammation were observed in model group Ⅱ rat liver, while the degree of hepatic injury was attenuated in the PGZ treatment group. Liver index, serum levels ofALT, AST, ALP, FINS and HOMA-IR were significantly increased （P〈0.05） in model group Ⅱ compared with control group Ⅱ. Whereas, in PGZ treatment group, serum levels of AST and FINS showed decreasing tendency, liver indexes, serum levels of ALT, ALP, TNF-α and HOMA-IR were significantly decreased compared with model group Ⅱ. Conclusion： Insulin resistance plays a role in the pathogenesis of NAFLD in rats. Pioglitazone can attenuate insulin resistance and biochemical and histological injury in high fat-induced fatty liver in rats.
基金supported by National Natural Science Foundation(No.81260315)Foundation of the Education Department of Guangxi Province,China(No.201010LX375)the Foundation of the Nature Science Fund,Guangxi Province,China(No.2012GXNSFBA053121)
文摘Objective： Previous studies have investigated the role of cytotoxic T-lymphocyte antigen-4 （CTLA-4） and tumor necrosis factor-alpha （TNF-a） in carcinogenesis of osteosarcoma, but their results were inconsistent. We aimed to clarify the associations between CTLA-4, TNF-a polymorphism and osteosarcoma risk by using meta-analysis. Methods： We searched relevant studies without language restriction in PubMed, EMbase, Cochrane Library, Google Scholar databases, Chinese National Knowledge Infrastructure （CNKI） and conference literature in humans published prior to March 2013. The strengths of the associations between genetic variants and osteosarcoma risk were estimated by odds ratio （OR） with 95% confidence interval （95% CI）. Results： A total of seven studies with 1,198 osteosarcoma patients and 1,493 controls were selected. Four studies were eligible for CTLA-4 （1,003 osteosarcoma and 1,162 controls）, and three studies for TNF-a （195 osteosarcoma and 331 controls）. Pooled results showed that rs231775 polymorphism of CTLA-4 was associated with osteosarcoma risk （GG vs. AA： OR=1.63, 95% CI=1.24-2.13; GG ＋ GA vs. AA： OR=1.56, 95% CI=1.21-2.01; AA ＋ GA vs. GG： OR=0.83, 95% CI=0.71-0.97; G vs. A： OR=1.21, 95% CI=1.08-1.36）. No significant heterogeneity was observed across the studies. No significant associations were found between rs5742909 polymorphism of CTLA-4 or rs1800629 polymorphism of TNF-a and osteosarcoma risk. Conclusions： These results suggest that the rs231775 polymorphism of CTLA-4 may play an important role in carcinogenesis of osteosarcoma.
基金funded by the National Key Research and Development Programs of China(No.2022YFC2603800)Jiangsu Provincial Key Research and Development Projects(No.BE2017693).
文摘Ticks are considered the second most common pathogen vectors transmitting a broad range of vital human and veterinary viruses.From 2017 to 2018,640 ticks were collected in eight different provinces in central and western China.Six species were detected,including H.longicornis,De.everestianus,Rh.microplus,Rh.turanicus,Rh.sanguineous,and Hy.asiaticum.Sixty-four viral metagenomic libraries were constructed on the MiSeq Illumina platform,resulting in 13.44 G(5.88×10^(7))of 250-bp-end reads,in which 2,437,941 are viral reads.We found 27 nearly complete genome sequences,including 16 genome sequences encoding entire protein-coding regions(lack of 30 or 50 end non-coding regions)and complete viral genomes,distributed in the arboviral family(Chuviridae,Rhabdoviridae,Nairoviridae,Phenuiviridae,Flaviviridae,Iflaviridae)as well as Parvoviridae and Polyomaviridae that cause disease in mammals and even humans.In addition,13 virus sequences found in Chuviridae,Nairoviridae,Flaviviridae,Iflaviridae,Hepeviridae,Parvoviridae,and Polyomaviridae were identified as belonging to a new virus species in the identified viral genera.Besides,an epidemiological survey shows a high prevalence(9.38%and 15.63%)of two viruses(Ovine Copiparvovirus and Bovine parvovirus 2)in the tick cohort.
文摘AIM To investigate and compare the analytical and clinical performance of Tian Long automatic hypersensitive hepatitis B virus(HBV) DNA quantification system and Roche CAP/CTM system.METHODS Two hundred blood samples for HBV DNA testing, HBV-DNA negative samples and high-titer HBV-DNA mixture samples were collected and prepared. National standard materials for serum HBV and a worldwide HBV DNA panel were employed for performance verification. The analytical performance, such as limit of detection, limit of quantification, accuracy, precision, reproducibility, linearity, genotype coverage and cross-contamination, was determined using the Tian Long automatic hypersensitive HBV DNA quantification system(TL system). Correlation and Bland-Altman plot analyses were carried out to compare the clinical performance of the TL system assay and the CAP/CTM system. RESULTS The detection limit of the TL system was 10 IU/m L, and its limit of quantification was 30 IU/m L. The differences between the expected and tested concentrations of the national standards were less than ± 0.4 Log10 IU/m L, which showed high accuracy of the system. Results of the precision, reproducibility and linearity tests showed that the multiple test coefficient of variation(CV) of the same sample was less than 5% for 102-106 IU/m L; and for 30-108 IU/m L, the linear correlation coefficient r2 = 0.99. The TL system detected HBV DNA(A-H) genotypes and there was no cross-contamination during the "checkerboard" test. When compared with the CAP/CTM assay, the two assays showed 100% consistency in both negative and positive sample results(15 negative samples and 185 positive samples). No statistical differences between the two assays in the HBV DNA quantification values were observed(P > 0.05). Correlation analysis indicated a significant correlation between the two assays, r2 = 0.9774. The Bland-Altman plot analysis showed that 98.9% of the positive data were within the 95% acceptable range, and the maximum difference was-0.49.CONCLUSION The TL system has good analytical performance, and exhibits good agreement with the CAP/CTM system in clinical performance.
基金supported by the Grant of Wuxi Technology Bureau Scientific and Technology Project (CSE31N1607)the Wuxi Key Medical Talents Program (ZDRC024)+5 种基金Shanghai Pujiang Program (15PJ1407300)the Wuxi Medical Development Discipline Program (FZXK006, MS201702)the Significant Program from Wuxi Health and Family Planning Commission (z201603)the National High Technology Research and Development Program of China (2014AA021403)the National Natural Science Foundation of China (81571977, 81701550)the Project from Shanghai Municipal Commission of Health and Family Planning (15GWZK0103)
文摘Little data is available on the evaluation of the occurrence rates of Epstein-Barr virus(EBV) in saliva and relationship with highly active antiretroviral therapy(HAART) use in HIV/AIDS patients in China. We conducted a retrospective cohort study of EBV serological tests for HIV/AIDS patients who were treated in the hospitals for infectious diseases in Wuxi and Shanghai, China from May 2016 to April 2017. The EBV-seropositive samples were identified by ELISA. EBV-specific primers and probes were used for the quantitative detection of viral DNA from saliva via quantitative real-time polymerase chain reaction. CD4 cell counts of the HIV/AIDS patients were detected by a flow cytometry. A total of 372 HIV/AIDS patients were ultimately selected and categorized for this retrospective cohort study. For EBV IgG and IgM, the HIV/AIDS HAART use(H) and non-HAART use(NH) groups had significantly higher seropositive rates than the HIV-negative control group. The HIV/AIDS(NH) group had the highest seropositive rate(IgG, 94.27%; IgM, 68.98%) and the highest incidence of EBV reactivation or infection. For salivary EBV DNA-positive rates and quantities, the HIV/AIDS(H)(73.69%) and the HIV/AIDS(NH)(100%) groups showed significantly higher values than the HIV-negative control group(35.79%,[ twofold). Further, the salivary EBV DNA-negative population had significantly higher CD4 cell counts than the EBV DNA-positive population in the HIV/AIDS(H) group and the HIV/AIDS(NH) groups. Thus, HAART use is beneficial in decreasing the EBV salivary shedding in HIV/AIDS patients and indirectly decreases EBV transmission risk.
基金supported by grants from the Key Natural Science Project of Anhui Provincial Education Department(Nos.KJ2018A0221,KJ2020A0578 and KJ2021A0773)National Innovation Program for College Students(No.202010367046)Anhui Provincial Undergraduate Innovative Training Program(Nos.S202010367008,S202110367030 and S202110367112).
文摘Columbianetin acetate(CE)is one of the effective components of Angelica pubescens.So far,the specific role and molecular mechanism of CE in pancreatic cancer are not clear.Thus,this study aimed to explore the specific mechanism of CE on pancreatic cancer.The target genes combined with CE were predicted through the PharmMapper database and the 3D molecular structure of CE.Then,the Cancer Genome Atlas(TCGA)and Cistrome data browser(DB)databases were used to screen Meiotic nuclear divisions 1(MND1)-related genes,transcription factors,and transcription factor data sets,and the intersection of the above data sets.The“limma”package in the R and gene expression profiling interactive analysis(GEPIA)databases were used to analyze the correlation and survival difference between the target genes and MND1 to predict the degree of association between CE and MND1.Western blotting and RT-PCR experiments revealed the regulatory relationship among CE,E2F1,and MND1 at the cellular level.The specific effects of CE on pancreatic cancer cells were explored through CCK8,wound healing,migration,and flow cycle experiments.E2F1,also the predictive transcription factor of MND1,was also the predictive target protein of CE.At the same time,E2F1 and MND1 were closely related in pancreatic tissue.In the cell function experiment,CE and interference with E2F1 expression could reduce the gene and protein expression of MND1,which was closely associated with cell proliferation,migration,and cycle development.Similarly,interfering with the expression of mnd1 can also inhibit the further development of tumor cells.CE may inhibit the development of pancreatic cancer cells by reducing the expression of MND1.This implies that CE may be a potential novel agent for the treatment of pancreatic cancer.
文摘To establish a rapid identification method for common pathogenic bacteria on the basis of molecular biology and to construct a preliminary Polymerase Chain Reaction-Capillary Electrophoresis - Restriction Fragment Length Polymorphism (PCR-CE-RFLP) database of bacteria isolated from clinical specimens frequently, 183 strains collected from clinical samples belonging to 12 genera and 19 species whose biochemical characterizations corresponded to the typical ones were examined. The genomic DNAs were amplified by two pairs of fluorescence labeled primers aiming at 16S rRNA gene and 16S-23S rRNA spacer region gene respectively at the same time. PCR products were then digested by restriction endonuclease HaeⅢ incompletely before taking capillary electrophoresis. The results with the PCR-CE-RFLP patterns of 16S rRNA genes were just alike within some genera, but when it comes to 16S-23S rRNA spacer region genes, each bacterium showed a unique pattern, which can be distinguished from each other easily. It seems that PCR-CE-RFLP patterns of 16S rRNA gene could only be used to classify the bacteria into family level, whereas the data of 16S-23S rRNA spacer region gene could be utilized to identify the whole microorganisms as precisely as the species level. In spite of the data of the spacer region gene alone can be sufficiently to verify the whole bacteria, we insist that the 16S rRNA gene could be of some assistant in case that there should be lots of families of bacteria, in which some similar ones, with the same RFLP data of 16S-23S rRNA spacer region gene, may coexist. This study proves that the utility of PCR-CE-RFLP is a convenient, rapid method to identify pathogenic bacteria, and is also a quick diagnosis measure for application to clinical use.
基金supported by the National Natural Science Foundation of China (Nos.22074015 and 22174014)。
文摘Developing accurate and sensitive DNA methyltransferase(MTase) analysis methods is essential for early clinical diagnosis and development of antimicrobial drug targets. In this work, by coupling WO_(3-x) dotsencapsulated metal-organic frameworks(MOFs) as co-reactants and terminal deoxynucleotidyl transferase(Td T)-mediated template-free branched polymerization, a dual signal-amplified electrochemiluminescent(ECL) biosensor was constructed to detect DNA adenine methylation(Dam) MTase. The employment of WO_(3-x) dots-encapsulated MOFs(i.e., NH_(2)-UIO66@WO_(3-x) ) was not only beneficial for biomolecule conjugation because of the abundant amino groups but also led to a 7-fold enhanced ECL response due to the increased loading of WO_(3-x). Moreover, Td T-mediated template-free branched polymerization promoted the capture of ECL emitters on the electrode surface, achieving 20-fold enhanced signal amplification. The presented ECL biosensor demonstrated a low detection limit of 2.4 × 10^(-4)U/m L, and displayed high reliability for the detection of Dam MTase in both spiked human serum and E. coli cell samples, and for the screening of potential inhibitors. This study opens a new avenue for designing a dual signal amplificationbased ECL bioassay for Dam MTase and screening inhibitors in the fields of clinical diagnosis and drug development.
基金the support from the National Natural Science Foundation of China (Nos.21974067, 22274073, 81971373 and 82001535)the National Key Research and Development Program of China (No.2018YFC1004700)+1 种基金Fundamental Research Funds for the Central Universities (Nos.020514380309,021414380502 and 2022300324)the State Key Laboratory of Analytical Chemistry for Life Science (Nos.5431ZZXM2305 and 5431ZZXM2204)。
文摘Spermatogenesis, maturation, capacitation and fertilization are precisely regulated by glycosylation. However, the relationship between altered glycosylation patterns and the onset and development of reproductive disorders is unclear, mainly limited by the lack of in situ imaging techniques for spermatozoa glycosylation. We developed an efficient and highly specific spermatozoa glycan imaging technique based on the robust chemoselective labeling of sialic acid(Sia) and N-acetyl-D-galactosamine(Gal/GalNAc). We further proposed a “tandem glycan chemoselective labeling” strategy to achieve simultaneous imaging of two types of glycans on spermatozoa. We applied the developed method to the spermatozoa from oligozoospermic patients and diabetic mice and found that these spermatozoa showed higher levels of Sia and Gal/Gal NAc expression than the normal groups. Moreover, spermatozoa from diabetic mice showed a severe decrease in number, viability, and forward motility, suggesting that in vivo glucose metabolism disorders may lead to an elevated level of spermatozoa glycosylation and have a correlation with the development of oligoasthenotspermia. Our work provides a research tool to reveal the relationship between glycosylation modification and spermatozoa quality, and a promising clue for the development of glycan-based reproductive markers.
基金supported by grants from the National Natural Science Foundation of China(No.81272372)Zhongnan Hospital of Wuhan University Science,Technology and Innovation Seed Fund(Wuhan,China)(No.PTXM2022017)The project was sponsored by SRF for ROCS,SEM.
文摘Posttranscriptional RNA modification is an important mode of epigenetic regulation in various biological and pathological contexts.N6,2′-O-dimethyladenosine(m6Am)is one of the most abundant methylation modifications in mammals and usually occurs at the first transcribed nucleotide.Accumulating evidence indicates that m6Am modifications have important roles in RNA metabolism and physiological and pathological processes.PCIF1(phosphorylated C-terminal domain interacting factor 1)is a protein that can bind to the phosphorylated C-terminal domain of RNA polymeraseⅡthrough its WW domain.PCIF1 is named after this binding ability.Recently,PCIF1 has been identified as a cap-specific adenine N6-methyltransferase responsible for m6Am formation.Discovered as the sole m6Am methyltransferase for mammalian mRNA,PCIF1 has since received more extensive and in-depth study.Dysregulation of PCIF1 contributes to various pathological processes.Targeting PCIF1 may hold promising therapeutic significance.In this review,we provide an overview of the current knowledge of PCIF1.We explore the current understanding of the structure and the biological characteristics of PCIF1.We further review the molecular mechanisms of PCIF1 in cancer and viral infection and discuss its therapeutic potential.
基金National Natural Science Foundation of China(Nos.81973408 and 82273445)1.3.5 Project for Disciplines of Excellence,West China Hospital,and Sichuan University(No.ZYYC20003)
文摘Metastasis-associated lung adenocarcinoma transcript 1(MALAT1)is a well-established oncogenic long non-coding RNA,the higher expression of which is strongly correlated with cancer events such as tumorigenesis,progression,metastasis,drug resistance,and treatment outcome in solid cancers.Recently,a series of studies has highlighted its potential role in hematological malignancies in terms of these events.Similar to solid cancers,MALAT1 can regulate various target genes via sponging and epigenetic mechanisms,but the miRNAs sponged by MALAT1 differ from those identified in solid cancers.In this review,we systematically describe the role and underlying mechanisms of MALAT1 in multiple types of hematological malignancies,including regulation of cell proliferation,metastasis,stress response,and glycolysis.Clinically,MALAT1 expression is related to poor treatment outcome and drug resistance,therefore exhibiting potential prognostic value in multiple myeloma,lymphoma,and leukemia.Finally,we discuss the evaluation of MALAT1 as a novel therapeutic target against cancer in preclinical studies.
基金supported by grants from the Natural Science Foundation of China(No.82072265)the National Natural Science Foundation of China(No.82302513)+3 种基金Guangdong Basic and Applied Basic Research Foundation(Nos.2022A1515110546,2024A1515013281)Science and Technology Project of Guangzhou(No.2023A03J0801)the Science and Technology Project of Guangzhou(No.202201020276)the Fund of Innovation and Entrepreneurship Leading Team Project of Guangzhou(No.201809010009).
文摘To the Editor:Current combined antiretroviral therapy(cART)can effectively suppress the plasma viral load in most people living with human immunodeficiency virus-1(HIV,PLWH).However,approximately 20-30%of PLWH undergoing cART can have very low but detectable plasma viral loads(<20 copies/mL,the limit of quantification in commercially available assays),known as residual HIV-1 low-level viremia(LLV).^([1])Residual HIV-1 LLV detection can reflect the source and dynamics of the HIV-1 reservoir in PLWH,thus playing an important role in both HIV-1 management and basic research.^([2])
基金supported by the National Natural Science Foundation of China[nos 82272411,82072363]Jiangsu Innovative and Entrepreneurial Talent Programme[JSSCBS20211602]+3 种基金PhD Research Startup Foundation of Affiliated Hospital of Nantong University[Tdb2011]Postdoctoral Research Foundation of Affiliated Hospital of Nantong University[BSH202110]Scientific Research Project of Nantong Municipal Health Commission[QN2022015],Nantong Basic Research Program[JC12022007]Grants from Jiangsu Provincial Research Hospital[YJXYY202204-YSC28].
文摘Background:Autophagy death of cancer cells is detrimental to apoptosis induced by therapeutic drugs,which promotes tumor progression to a certain extent.Increasing reports have demonstrated the regulatory role of circular RNAs(circRNAs)in autophagy.Here,we aimed to determine the role of hsa_circ_0009109 in autophagy in gastric cancer(GC).Methods:The effects of hsa_circ_0009109 on autophagy were examined using quantitative real-time polymerase chain reaction(qPCR),transmission electron microscopy,Western blot,and immunofluorescence.The mechanism of hsa_circ_0009109 regulating the miR-544a-3p/bcl-2 axis was analysed using fluorescence in situ hybridization,dual-luciferase reporter,and rescue experiments.Results:Functional testing indicated that hsa_circ_0009109 was significantly down-expressed in GC tissues and cell lines.A reduction in cytoplasmic-derived hsa_circ_0009109 could promote GC progression by accelerating cell proliferation,enhancing migration and invasion,inhibiting apoptosis,and accelerating the cell cycle progression.Besides,hsa_circ_0009109 was found to exert the effect of an autophagy inhibitor such as 3-Methyladenine(3-MA),which was manifested by the weakening of the immunofluorescence of LC3B and the reduction in autophagy-related proteins after overexpression of hsa_circ_0009109,while increased autophagosomes were observed after interference with hsa_circ_0009109.Subsequently,the crosstalk between hsa_circ_0009109 and miR-544a-3p/bcl-2 was verified using dual-luciferase reporter assay.The autophagy status was altered under the regulation of the hsa_circ_0009109-targeted miR-544a-3p/bcl-2 axis.Conclusions:The hsa_circ_0009109 mediated a novel autophagy regulatory network through targeting the miR-544a-3p/bcl-2 axis,which may shed new light on the exploration of therapeutic targets for the clinical treatment of GC.
基金supported by the National Natural Science Foundation of China Programs(81974382,82072068,82173315,82072167 and 82272277)the Technical Innovation Major Foundation Program of Hubei Province(2022BCA013)+1 种基金the National Key R&D Program of China(2022YFC2408100)Hubei Province Science and Technology Innovation Team Project([2022]No.11).
文摘Background:Metastasis is the main cause of death in colorectal cancer(CRC).Metastasis is a sequential and dynamic process,but the development of tumor cells during this process is unclear.In this study,we aimed to reveal characteristics of tumor cell subset during CRC metastasis.Methods:Single-cell RNA sequence CRC data of normal epithelium,non-metastatic primary tumor,metastatic primary tumor,and liver metastases from gene expression omnibus(GEO)dataset were analyzed to reveal characteristics of CRC metastasis.Primary tu-mor tissues of three non-metastatic CRC and three metastatic CRC patients from Union Hospital of Tongji Medical College(Wuhan,China)were used to verify the characteristics of CRC metastasis.Results:We identified a metastasis-related cancer cell subset EP1,which was characterized with a high expression of KRT17,LAMC2,EMP1,and PLAC8.EP1 had an enhanced cell-cell interaction,which interacted with SPPþmacrophages and drove them toward anti-inflammatory and immunosuppressive phenotype.Dynamic changes in genes and TF regulons during the metastasis were also revealed.Conclusions:This study advanced our understanding of the development of tumor cells during CRC metastasis and further identi-fied metastasis-related subset and potential therapeutic targets for the treatment and prevention of CRC metastasis.
基金supported by the National Natural Science Foundation of China Programs(82072068,82072167,82472135,82272277,and 82173315)Natural Science Foundation of Hubei Province(2024AFA053)+4 种基金the Major Scientific and Technological Innovation Projects in Hubei Province(2022BCA013)the Union Hospital Foun-dation for Young Scientist(2021xhqh01)the Knowledge Innovation Special Project for Fundamental Research of Wuhan(2022020801010461)Hubei Province Science and Technology Inno-vation Team Project([2022]No.11)the Open Foundation of Hubei Key Laboratory of Regenerative Medicine and Multi-disciplinary Translational Research(2022zsyx001).
文摘Hypoxia,a prevalent characteristic of solid tumors,substantially impairs the efficacy of cancer treatments.However,there are no feasible clinical approaches for treating hypoxic tumors.Here,we develop metal-phenolic networks(CuGI)utilizing the natural glycolysis inhibitor(epigallocatechin gallate)and the essential metal element in the human body(copper ions),specifically targeting and annihilating hypoxic cancer cells.CuGI redirects the metabolic pathway of hypoxic cancer cells from anaerobic glycolysis to oxidative phosphorylation,thereby enhancing reactive oxygen species production and promoting oligomerization of lipoylated proteins in the tricarboxylic acid cycle.Through targeted induction of oxidative and proteotoxic stresses,CuGI induces apoptosis and cuproptosis specifically in cancer cells under hypoxic conditions while sparing normal cells.Moreover,cancer cell membrane-coated CuGI(CuGI@CM)exhibits enhanced tumor penetration effect and demonstrates commendable biocompatibility,effectively suppressing colorectal tumor growth.Importantly,CuGI@CM,when combined with vascular disruptors or radiotherapy which aggravate tumor hypoxia,syner-gistically potentiates therapeutic efficacy.Thus,CuGI represents a specific and potent nanotherapeutic capable of selectively eliminating hypoxic tumors,offering promise in combination therapies to address tumor hypoxia.
基金This study was supported by grants from the National Natural Science Foundation of China （No. 30972819） and Program of Beijing Science and Technology Activity for Scholars Abroad （No. LXZZ2008008）.
文摘Background The normal microbial flora of the vagina plays an important role in preventing genital and urinary tract infections in women. Thus an accurate understanding of the composition and ecology of the ecosystem is important to understanding the etiology of these diseases. This study aimed to compare the characteristics of main lactobacillus species between healthy women and women with bacterial vaginosis （BV） by quantitative culture and PCR methods. Main lactobacillus species include L. crispatus , L. gasseri, L. jensenii and L. iners.Methods A total of 150 Women attending Gynecology Outpatient Clinic of Beijing Friendship Hospital, were diagnosed as having BV because three or more of the following criteria were met （standard of Amsel＇s composite criteria）： homogeneous discharge, elevated vaginal pH （pH 〉4.5）, production of amines, and presence of ＂clue＂ cells. Those with less than three of the criteria were considered as healthy. Simultaneously, smears were made of vaginal fluid and Gram stained, then were assessed qualitatively as normal （grade Ⅰ）, intermediate （grade Ⅱ）, or consistent with BV （grade Ⅲ）. Gardnerella vaginalis were identified by using Vitek 2 Compact and PCR methods. Lactobacillus species were identified by PCR methods. Gardnerella vaginalis and lactobacilli colony counts were determined by calculating the most number of colonies of each species in the appropriate plates （colonies between 10 and 300）, corrected by the dilution of the sample in the plates, and multiplied by 10 （to account for plating 100 μl）, in order to get colony forming units per milliliter of vaginal secretion.Results BV was diagnosed in 31% （46/150） patients using the composite criteria, the remainder being regarded as healthy. The majority of patients with BV had a smear assessed as grade Ⅲ （91%, 42/46） and minority of them had a smear assessed as grade Ⅱ（9%, 4/46）. The majority of healthy women had a smear assessed as grade Ⅰ （64%, 67/104）. Smears assessed as grade Ⅱwere found （36%, 37/104） among patients diagnosed as healthy, with less than three of the composite criteria. L. crispatus was cultured from 94% of healthy women and 83% of women with BV, with the former colonies count average value of 106 and the latter of 103. L. gasseri, L. iners, and L. jensenfi were cultured from 85%, 68% and 43% of healthy women; and 28%, 89% and 44% of BV women, respectively.Conclusions The quantities of four lactobacillus species except L. jensenii had a significant difference between healthy women and women with BV. Our results provide support for the negative association between L. iners and L. gasseri. Although L. crispatus were existent both in healthy and BV positive women＇s vagina, the numbers of L. crispatus were significantly different for the dominant number in healthy women. Smears of vaginal fluid and Gram stain play an important guiding role in bacteria culture.
