This study investigated the mechanism underlying electroacupuncture therapy for vascular dementia through electroacupuncture at the acupoints of Baihui （DU20）, Dazhui （DU14）, and bilateral Shenshu （BL23） in a ra...This study investigated the mechanism underlying electroacupuncture therapy for vascular dementia through electroacupuncture at the acupoints of Baihui （DU20）, Dazhui （DU14）, and bilateral Shenshu （BL23） in a rat model of vascular dementia produced by bilateral middle cerebral artery occlusion. Morris water maze test showed that electroacupuncture improved the learning ability of vascular dementia rats. Western blot assay revealed that the expression of p70 ribosomal protein S6 kinase and ribosomal protein S6 in vascular dementia rats was significantly increased after electroacupuncture, compared with the model group that was not treated with acupuncture. The average escape latency was also shortened after electroacupuncture, and escape strategies in the spatial probe test improved from edge and random searches, to linear and trending swim pathways. The experimental findings indicate that electroacupuncture improves learning and memory ability by up-regulating expression of p70 ribosomal protein S6 kinase and ribosomal protein S6 in the hippocampus of vascular dementia rats.展开更多
BACKGROUND Systemic lupus erythematosus(SLE)is an autoimmune disease characterized by systemic involvement and multiple autoantibodies in the serum.Patients with protein C(PC)and protein S(PS)deficiency are prone to t...BACKGROUND Systemic lupus erythematosus(SLE)is an autoimmune disease characterized by systemic involvement and multiple autoantibodies in the serum.Patients with protein C(PC)and protein S(PS)deficiency are prone to thrombosis.In contrast,patients with primary hyperfibrino-lysis tend to bleed.CASE SUMMARY A 52-year-old female patient with bilateral pleural effusion was diagnosed with"tuberculous pleurisy"and treated with anti-tuberculosis drugs and prednisone.The coagulation-related laboratory results showed decreased fibrinogen,PC activity,PS activity,and antithrombinШactivity.The immune-related laboratory results showed positive antinuclear antibody,anti-Smith antibody,anticardiolipin antibody(ACL),anti-β2-glycoprotein I antibody(aβ2GPI)and direct Coomb’s test and decreased complement 3 and complement 4.Thoracoscopy was performed and bloody pleural fluid was drained.Pathology of the pleural biopsy showed lymphocytes,plasma cells,and a few eosinophils in adipose and fibrous connective tissue.Results of whole exome sequencing of blood showed no genetic mutations suggesting the presence of hereditary hematological diseases.The patient was finally diagnosed with SLE and primary hyperfibrinolysis,and was treated with prednisolone,hydroxychloroquine,and compound cyclophosphamide.CONCLUSION PC and PS deficiency in SLE might be related to ACL and aβ2GPI.SLE and primary hyperfibrinolysis can coexist in one patient,with both a risk of thrombosis and a risk of bleeding.展开更多
The ribosomal protein S29 also known as RPS29, is not only a component of the 40S subunit of ribosome, but also involved in embryonic development, oncogenesis and other pathologic conditions. However, rare commercial ...The ribosomal protein S29 also known as RPS29, is not only a component of the 40S subunit of ribosome, but also involved in embryonic development, oncogenesis and other pathologic conditions. However, rare commercial antibody against RPS29 restricts the discovery of precise physiological and pathological function of this protein. In this study, the whole RPS29 gene was inserted into plasmid pGEX-6p-1 to express glutathione＇s transferase （GST） fusion proteins in Escherichia eoli （E. coli） strain BL21. High yields of soluble recombinant proteins were obtained. Mice were immunized with the recombinant RPS29 protein. The serum from the immunized mice could specially react with purified recombinant RPS29 proteins and native RPS29 proteins in CCE cells by western blotting, immunofluorescence staining and flow cytometric analysis. Further more the polyclonal antibodies also reacted specifically with human cell strain ECV304, which showed typical cytoplasmatic fluorescence. The polyclonal antibodies we prepared would be an available tool for studying the roles of RPS29 in embryonic development and human diseases.展开更多
A 76-year-old woman was diagnosed as having left lilac deep vein thrombosis due to a hereditary deficiency of protein S, seventeen members of her family were studied with the measurements of total protein S (TPS), fre...A 76-year-old woman was diagnosed as having left lilac deep vein thrombosis due to a hereditary deficiency of protein S, seventeen members of her family were studied with the measurements of total protein S (TPS), free protein S (FPS), protein C (PC) and anti-thrombin-Ⅲ(AT-Ⅲ). The results showed that the level of FPS of thepatient was only 7%, while that of TPS 137.1%. Her secondson had a low FPS level (13.4%) too, and his TPS level was 48.1%. PC and AT-Ⅲwere all normal It was thus the first case of hereditary PS dificiency reported in China.展开更多
The Tyro3, Axl and Mer (TAM) receptor tyrosine kinases are activated by endogenous ligands, protein S1 (PROS1) and growth arrest-specific gene 6 (Gas6), and those have important effects on cell biology. These receptor...The Tyro3, Axl and Mer (TAM) receptor tyrosine kinases are activated by endogenous ligands, protein S1 (PROS1) and growth arrest-specific gene 6 (Gas6), and those have important effects on cell biology. These receptors (Rs) can be shad from the cell membrane and their soluble(s) forms can be found in plasma. We investigated the fluctuation and interactive role of sTAMRs and its ligands in patients with hepatocellular carcinoma (HCC), hepatitis groups, and healthy normal adult controls (NC). The measurement cases were 45 patients with HCC group (stage 1 in 4, stage 2 in 8, stage 3 in 16, and stage 4 in 17), 4 patients with fulminant hepatitis (FH), 14 patients with acute hepatitis (AH), 10 patients with chronic hepatitis (CH), 16 patients with liver cirrhosis (LC), and 20 NCs matched by age. Plasma levels of three sTAMRs and their ligands were measured by ELISA. In comparison with NCs, Gas6, des-γ-carboxy Gas6, and sTAMRs levels were significantly higher in HCC patients, but free PROS1 levels were significantly lower. The sTyro3 and sAxl levels peaked HCC stages 2 and 3 respectively, and gradually decreased afterwards while maintaining high levels. sMer levels increased with the progression of HCC. Gas6 and des-γ-carboxy Gas6 levels gradually increased, and PROS1 levels decreased with the progression of HCC. Gas6 levels were positively correlated with sAxl levels, whereas sMer levels were negatively correlated with free PROS1 levels. sTAMRs and Gas6 levels increased in parallel to the progression of HCC fibrosis. Through the progression of HCC, Axl played the major role in TAMRs activation. However, sTYro3 continued increasing rapidly from the early stage, and that of Mer increased throughout the progression. Roles of Axl may be changed in Mer, because des-γ-carboxy Gas6 levels increasing with Gas6 in the advanced stage of HCC cannot send a signal to Axl.展开更多
PC and PS antigens were measured in 370 cases of 12 different diseases and 14 women at third trimester of pregnancy. The PC level was found significantly raised in coronary heart disease, diabetes, nephrotic syndrome,...PC and PS antigens were measured in 370 cases of 12 different diseases and 14 women at third trimester of pregnancy. The PC level was found significantly raised in coronary heart disease, diabetes, nephrotic syndrome, stroke, aplastic anemia and the third trimester of pregnancy, decreased in chronic hepatitis, acute fulminant hepatitis, cirrhosis and deep vein thrombosis (DVT) and no change in acute hepatitis, primary liver carcinoma, Buerger’s disease and leukemia. Total PS was increased in type Ⅱdiabetes, acute lymphocytic leukemia and DVT, and decreased in Berger’s disease, liver diseases, aplastic anemia and the third trimester of pregnancy. In majority of the cases, the total PS and free PS changed parallelly except in Buerger’s disease, DVT and preeclampsia. No correlation was found between PC and PS, PC and AT-Ⅲ.展开更多
To editor:Thrombophilia is a group of blood clotting disorders that increase the risk of venous thromboembolism(VTE),which is primarily caused by genetic mutations in proteins involved in the coagulation cascade.These...To editor:Thrombophilia is a group of blood clotting disorders that increase the risk of venous thromboembolism(VTE),which is primarily caused by genetic mutations in proteins involved in the coagulation cascade.These mutations include Factor V Leiden,prothrombin G20210A,protein S deficiency(PSD),antithrombin III deficiency,and methyltetrahydrofolate reductase.Among these,PSD contribute to a significant proportion of thromboembolic events in pregnant women,particularly in those with additional risk factors such as obesity,advanced maternal age,or a history of VTE.1 This correspondence aims to present a series of clinical cases of PSD in pregnant women,highlighting treatment strategies,especially in a hospital in a low-and middle-income country(LMIC)with limited access to advanced therapeutic options.Written informed consent was obtained from all patients involved in this study.We had treated six pregnant women diagnosed with PSD.Among these cases,four had a history of adverse obstetric outcomes,including abortion,preterm birth,or intrauterine fetal death,while the other two cases had recurrent miscarriages(Supplementary Table 1,http://links.lww.com/MFM/A72).These women were identified after the diagnosis of PSD,and each case was carefully monitored throughout pregnancy.展开更多
BACKGROUND Upper gastrointestinal hemorrhage is a life-threatening manifestation of cow’s milk protein allergy(CMPA).We analyze the clinical characteristics of a case of milk protein allergy manifested as severe uppe...BACKGROUND Upper gastrointestinal hemorrhage is a life-threatening manifestation of cow’s milk protein allergy(CMPA).We analyze the clinical characteristics of a case of milk protein allergy manifested as severe upper gastrointestinal hemorrhage.CASE SUMMARY The hospital admitted a 2-month-old male infant due to“melena for 6 days,he-matemesis twice”.The main symptom was melena,initially occurring once or twice per day,then gradually increasing to five or six times per day at their peak.During the course of the illness,the infant vomited blood,but there were no re-ports of vomiting,fever,pale complexion,dyspnea,wheezing,or difficulty brea-thing.Laboratory tests showed hemoglobin level of 87 g/L,platelet count of 349×109/L,and eosinophil percentage of 0.031.Coagulation studies were normal.After avoiding certain foods and feeding with an amino acid formula for 2 weeks,a repeat gastroscopy revealed less bleeding.After six weeks,a positive oral food challenge test confirmed a severe CMPA.At the 4-month follow-up,there was no gastrointestinal bleeding,and the infant was growing and developing well.CONCLUSION The manifestations of milk protein allergy are diverse and nonspecific,with gas-trointestinal bleeding being less common,especially in infants.When infants present with unexplained massive hematemesis,it’s critical to investigate the possibility of CMPA.展开更多
Objective: To analyze the dynamic change of serum protein S100b in patients with traumatic brain injury and its clinical value in assessing brain damage. Methods: According to Glasgow coma scale (GCS), 102 cases of tr...Objective: To analyze the dynamic change of serum protein S100b in patients with traumatic brain injury and its clinical value in assessing brain damage. Methods: According to Glasgow coma scale (GCS), 102 cases of traumatic brain injury were divided into mild brain injury group (GCS≥13, n=31, Group A), moderate brain injury group (8<GCS<13, n=37, Group B) and severe brain injury group (GCS≤8, n=34, Group C). Serial S100b concentrations were analyzed by enzyme-linked immunosorbent assay (ELISA) in blood samples taken on admission, 12 h, 24 h, 48 h, 72 h and 7 days after traumatic brain injury. Results: The severe brain injury group showed significantly higher concentration of serum S100b, with earlier increase and longer duration, than the mild and moderate brain injury groups. The patients with higher S100b exhibited lower GCS scores and poor clinical prognosis. The increase in S100b could emerge before clinical image evidence indicated so. Conclusions: Serum S100b can be used as a sensitive index for assessment and prediction of traumatic brain injury severity and prognosis.展开更多
Objective： To study the effects of Yiqi Huoxue Recipe （益气活血方） and Coxsackie B virus type 3 （CVB3） on the expression of ribosomal protein S20 in rat cardiac myocytes, to explore the pathogenesis of myocarditi...Objective： To study the effects of Yiqi Huoxue Recipe （益气活血方） and Coxsackie B virus type 3 （CVB3） on the expression of ribosomal protein S20 in rat cardiac myocytes, to explore the pathogenesis of myocarditis induced by CVB3 and the mechanism of Yiqi Huoxue Recipe on gene level, and to further investigate whether Yiqi Huoxue Recipe is an effective prescription for CVB3 myocarditis. Methods： A modified suppression subtractive hybridization （SSH） was used to isolate differentially expressed genes between the CVB3 infection group and the treatment group with Yiqi Huoxue Recipe. The results were further verified by fluorescence RT-PCR. Results： The results of SSH showed that the gene expression of ribosomal protein S20 in the treatment group was higher than that in the CVB3 infection group （P〈0.05）, which agreed with the results of fluorescent RT-PCR. Conclusion： Down-regulation of ribosomal protein S20 mRNA expression might be one of the mechanisms in CVB3 myocarditis, and Yiqi Huoxue Recipe could achieve the treatment of viral myocarditis by regulating the expression of ribosomal protein S20.展开更多
Ribosomal protein S 13 gene has been cloned and analyzed in many organisms,but there are few documents relating to insects. In this communication, the full-length cDNA sequence of ribosomal protein S 13 gene in the di...Ribosomal protein S 13 gene has been cloned and analyzed in many organisms,but there are few documents relating to insects. In this communication, the full-length cDNA sequence of ribosomal protein S 13 gene in the diamondback moth, Plutella xylostella(Lepidoptera: Plutellidae), was determined by using PCR amplification technique. The features of the ribosomal protein S 13 gene sequence were analyzed and the deduced amino acids sequence was compared with those from other insects. The results of multi-alignment of the amino acid sequences between the diamondback moth and other insect species revealed that this gene sequence is highly conserved in insects. Based on maximum likelihood method, a phylogenetic tree was constructed from 10 different species using PHYLIP software. It showed that nematode is one separate lineage and the five insect speciesbe long to another lineage, whereas those species higher than insects form the third one. The pattern of this phylogenetic tree evidently represented the evolution of different species.展开更多
Objective To investigate the mechanism of anticoagulation protein defect in the pathogenesis of unexplained recurrent miscarriage. Methods Fifty-seven patients with a history of unexplained abortion were enrolled as t...Objective To investigate the mechanism of anticoagulation protein defect in the pathogenesis of unexplained recurrent miscarriage. Methods Fifty-seven patients with a history of unexplained abortion were enrolled as the investigation group for tests of protein C, protein S, antithrombinⅢ(AT-Ⅲ), as well as activated protein C resistance (APC-R). The control group con-sisted of fifty healthy women with a history of normal pregnancy and delivery. Blood samples were obtained for measuring serum activity of protein C, protein S, AT-Ⅲ, and APC-R. Patients with positive APC-R were tested for factorⅤ(FⅤ) Lei-den gene mutation by PCR-RFLP method. Results Of the 57 patients, 12 (21.1%), 1 (1.8%), and 5 (8.8%) cases were found with protein S, protein C, and AT-Ⅲdeficiency respectively, and 13 (22.8%) cases with positive results of APC-R. Of the control group, no protein C or AT-Ⅲdeficiency was ever found, whereas 2 (4.0%) volunteers were presented with protein S deficiency and 3 (6.0%) with positive results of APC-R. No FⅤLeiden gene mutation was identified in all the patients with positive APC-R results. Late spontan-eous abortion cases had higher incidence of anticoagulation protein defect than the early cases. Conclusion Anticoagulation protein defect may play a role in the pathogenesis of fetal loss, especially for those occurr-ing in late stage of pregnancy.展开更多
Previous results in this laboratory indicated that in ribosomal protein S12 strepto-mycin-dependent mutants of Bacillus subtilis, the burst size of bacteriophage φ105 was decreased, and the protein synthesis was inhi...Previous results in this laboratory indicated that in ribosomal protein S12 strepto-mycin-dependent mutants of Bacillus subtilis, the burst size of bacteriophage φ105 was decreased, and the protein synthesis was inhibited, while the DNA and展开更多
[Objective] ＂Tapping panel dryness （TPD）＂, a syndrome known as tapping incision blocked partly or entirely during latex exploiting, has become the most important factor causing great losses for rubber production. ...[Objective] ＂Tapping panel dryness （TPD）＂, a syndrome known as tapping incision blocked partly or entirely during latex exploiting, has become the most important factor causing great losses for rubber production. Aiming to elucidate the molecular mechanism of tapping panel dryness occurrence, this study carried out molecular cloning and bioinformatical analysis of a mRPL21 cDNA sequence, a gene associated with TPD. [Method] In a preliminary study, an expressed sequence tag （EST） encoding a deduced protein homologous to mitochondrial 50S ribosomal protein L21 （mRPL21）, which showed to be down-regulated in the latex of TPD-affected rubber trees, was isolated by suppression subtractive hybridization （SSH）. After ESTs assembling and RT-PCR validation, an 853 bp cDNA sequence with an open reading frame （ORF） was cloned, which was named as HbmRPL21 under GenBank accession number of HM230670. [Result] Bioinformatical analysis suggests that HbmRPL21 encodes a deduced polypeptide of 271 amino acids with a theoretical molecular weight （Mw） of 30.52 kDa and isolectric point （pI） of 8.40, and HbmRPL21 is a mitochondrion-targeted protein with a conserved domain of Ribosomal_L21p involving translation. Homology analysis reveals high amino acid sequence identity of mRPL21 from plants, while diversity of that between plant and animal kingdom. [Conclusion] This study laid the basis for further revealing the biological functions of mRPL21 in TPD-affected rubber trees.展开更多
BACKGROUND Tacrolimus(FK506)is a key calcineurin inhibitor used to prevent organ transplant rejection and is effective in improving graft survival.However,it is linked to hyperglycemia and insulin resistance,contribut...BACKGROUND Tacrolimus(FK506)is a key calcineurin inhibitor used to prevent organ transplant rejection and is effective in improving graft survival.However,it is linked to hyperglycemia and insulin resistance,contributing to new-onset diabetes after transplantation and negatively affecting islet function.AIM To study the effects of tacrolimus on the insulin signaling pathway of hepatocytes.METHODS HL7702 cells were treated with different concentrations of tacrolimus(0.1 mg/L,1 mg/L,5 mg/L)for 24 hours.The proteins involved in insulin signaling were detected by Western blotting.RESULTS Compared with the control group,phosphorylation of insulin receptor substrate(IRS)1 at Ser 307 and Ser 323 were increased significantly when the tacrolimus concentration reached 1 and 5 mg/L.Phosphorylation of IRS1 at Ser 1101 was also increased,although not significantly.However,phosphorylation of Ribosomal protein S6 kinase beta-1 at Thr 389 was decreased significantly.The levels of phosphorylated glycogen synthase kinase 3αSer 21 and Ser 9 were increased.Surprisingly,phosphorylation of glycogen synthase at Ser 641 was increased.There was no significant change in the activity of glycogen phosphorylase.CONCLUSION Tacrolimus has no direct effect on hepatic glucose metabolism,but inhibits IRS1-mediated insulin signaling.This may be one of the underlying mechanisms by which tacrolimus induces insulin resistance.展开更多
AIM: To investigate the transactivating effect of complete S protein of hepatitis B virus (HBV) and to construct a subtractive cDNA library of genes transactivated by complete S protein of HBV by suppression subtracti...AIM: To investigate the transactivating effect of complete S protein of hepatitis B virus (HBV) and to construct a subtractive cDNA library of genes transactivated by complete S protein of HBV by suppression subtractive hybridization (SSH) technique and to clone genes associated with its transactivation activity, and to pave the way for elucidating the pathogenesis of hepatitis B virus infection. METHODS: pcDNA3.1(-)-complete S containing full-length HBV S gene was constructed by insertion of HBV complete S gene into BamH I/Kpn I sites. HepG2 cells were cotransfected with pcDNA3.1(-)-complete S and pSV-lacZ. After 48 h, cells were collected and detected for the expression of β-galactosidase (β-gal). Suppression subtractive hybridization and bioinformatics techniques were used. The mRNA of HepG2 cells transfected with pcDNA3.Incomplete S and pcDNA3.1(-) empty vector was isolated, and detected for the expression of complete S protein by reverse transcription polymerase chain reaction (RT-PCR) method, and cDNA was synthesized. After digestion with restriction enzyme RsaI, cDNA fragments were obtained. Tester cDNA was then divided into two groups and ligated to the specific adaptors 1 and 2, respectively. After tester cDNA had been hybridized with driver cDNA twice and underwent nested PCR twice, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. Amplification of the library was carried out within E. coli strain DH5α. The cDNA was sequenced and analyzed in GenBank with BLAST search after polymerase chain reaction (PCR) amplification. RESULTS: The complete S mRNA could be detected by RT-PCR in HepG2 cells transfected with the pcDNA3.1(-)-complete S. The activity of β-gal in HepG2 cells transfected with the pcDNA3.1(-)-complete s was 6.9 times higher than that of control plasmid. The subtractive library of genes transactivated by HBV complete S protein was constructed successfully. The amplified library contains 86 positive clones. Colony PCR showed that 86 clones contained DNA inserts of 200-1 000 bp, respectively. Sequence analysis was performed in 35 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 33 coding sequences were obtained. These cDNA sequences might be target genes transactivated by complete S protein of HBV. Moreover, two unknown genes were discovered, full length coding sequences were obtained by bioinformatics techniques, one of them was named complete S transactivated protein 1 (CSTP1) and registered in GenBank (AY553877). CONCLUSION: The complete S gene of HBV has a transactivating effect on SV40 early promoter. A subtractive cDNA library of genes transactivated by HBV complete S protein using SSH technique has been constructed successfully. The obtained sequences may be target genes transactivated by HBV complete S protein among which some genes coding proteins are involved in cell cycle regulation, metabolism, immunity, signal transduction, cell apoptosis and formation mechanism of hepatic carcinoma.展开更多
基金supported by the National Natural Science Foundation of China, No. 81001541
文摘This study investigated the mechanism underlying electroacupuncture therapy for vascular dementia through electroacupuncture at the acupoints of Baihui （DU20）, Dazhui （DU14）, and bilateral Shenshu （BL23） in a rat model of vascular dementia produced by bilateral middle cerebral artery occlusion. Morris water maze test showed that electroacupuncture improved the learning ability of vascular dementia rats. Western blot assay revealed that the expression of p70 ribosomal protein S6 kinase and ribosomal protein S6 in vascular dementia rats was significantly increased after electroacupuncture, compared with the model group that was not treated with acupuncture. The average escape latency was also shortened after electroacupuncture, and escape strategies in the spatial probe test improved from edge and random searches, to linear and trending swim pathways. The experimental findings indicate that electroacupuncture improves learning and memory ability by up-regulating expression of p70 ribosomal protein S6 kinase and ribosomal protein S6 in the hippocampus of vascular dementia rats.
文摘BACKGROUND Systemic lupus erythematosus(SLE)is an autoimmune disease characterized by systemic involvement and multiple autoantibodies in the serum.Patients with protein C(PC)and protein S(PS)deficiency are prone to thrombosis.In contrast,patients with primary hyperfibrino-lysis tend to bleed.CASE SUMMARY A 52-year-old female patient with bilateral pleural effusion was diagnosed with"tuberculous pleurisy"and treated with anti-tuberculosis drugs and prednisone.The coagulation-related laboratory results showed decreased fibrinogen,PC activity,PS activity,and antithrombinШactivity.The immune-related laboratory results showed positive antinuclear antibody,anti-Smith antibody,anticardiolipin antibody(ACL),anti-β2-glycoprotein I antibody(aβ2GPI)and direct Coomb’s test and decreased complement 3 and complement 4.Thoracoscopy was performed and bloody pleural fluid was drained.Pathology of the pleural biopsy showed lymphocytes,plasma cells,and a few eosinophils in adipose and fibrous connective tissue.Results of whole exome sequencing of blood showed no genetic mutations suggesting the presence of hereditary hematological diseases.The patient was finally diagnosed with SLE and primary hyperfibrinolysis,and was treated with prednisolone,hydroxychloroquine,and compound cyclophosphamide.CONCLUSION PC and PS deficiency in SLE might be related to ACL and aβ2GPI.SLE and primary hyperfibrinolysis can coexist in one patient,with both a risk of thrombosis and a risk of bleeding.
基金Supported by the National Natural Science Foundation of China(30800983,30700418 and 30972596)the Natural Science Foundation of Chongqing(2008BB5113 and 2009BB5015) the Scientific Research Foundation of Third Military Medical University(2009XHG03 and 2009XYY04)
文摘The ribosomal protein S29 also known as RPS29, is not only a component of the 40S subunit of ribosome, but also involved in embryonic development, oncogenesis and other pathologic conditions. However, rare commercial antibody against RPS29 restricts the discovery of precise physiological and pathological function of this protein. In this study, the whole RPS29 gene was inserted into plasmid pGEX-6p-1 to express glutathione＇s transferase （GST） fusion proteins in Escherichia eoli （E. coli） strain BL21. High yields of soluble recombinant proteins were obtained. Mice were immunized with the recombinant RPS29 protein. The serum from the immunized mice could specially react with purified recombinant RPS29 proteins and native RPS29 proteins in CCE cells by western blotting, immunofluorescence staining and flow cytometric analysis. Further more the polyclonal antibodies also reacted specifically with human cell strain ECV304, which showed typical cytoplasmatic fluorescence. The polyclonal antibodies we prepared would be an available tool for studying the roles of RPS29 in embryonic development and human diseases.
文摘A 76-year-old woman was diagnosed as having left lilac deep vein thrombosis due to a hereditary deficiency of protein S, seventeen members of her family were studied with the measurements of total protein S (TPS), free protein S (FPS), protein C (PC) and anti-thrombin-Ⅲ(AT-Ⅲ). The results showed that the level of FPS of thepatient was only 7%, while that of TPS 137.1%. Her secondson had a low FPS level (13.4%) too, and his TPS level was 48.1%. PC and AT-Ⅲwere all normal It was thus the first case of hereditary PS dificiency reported in China.
文摘The Tyro3, Axl and Mer (TAM) receptor tyrosine kinases are activated by endogenous ligands, protein S1 (PROS1) and growth arrest-specific gene 6 (Gas6), and those have important effects on cell biology. These receptors (Rs) can be shad from the cell membrane and their soluble(s) forms can be found in plasma. We investigated the fluctuation and interactive role of sTAMRs and its ligands in patients with hepatocellular carcinoma (HCC), hepatitis groups, and healthy normal adult controls (NC). The measurement cases were 45 patients with HCC group (stage 1 in 4, stage 2 in 8, stage 3 in 16, and stage 4 in 17), 4 patients with fulminant hepatitis (FH), 14 patients with acute hepatitis (AH), 10 patients with chronic hepatitis (CH), 16 patients with liver cirrhosis (LC), and 20 NCs matched by age. Plasma levels of three sTAMRs and their ligands were measured by ELISA. In comparison with NCs, Gas6, des-γ-carboxy Gas6, and sTAMRs levels were significantly higher in HCC patients, but free PROS1 levels were significantly lower. The sTyro3 and sAxl levels peaked HCC stages 2 and 3 respectively, and gradually decreased afterwards while maintaining high levels. sMer levels increased with the progression of HCC. Gas6 and des-γ-carboxy Gas6 levels gradually increased, and PROS1 levels decreased with the progression of HCC. Gas6 levels were positively correlated with sAxl levels, whereas sMer levels were negatively correlated with free PROS1 levels. sTAMRs and Gas6 levels increased in parallel to the progression of HCC fibrosis. Through the progression of HCC, Axl played the major role in TAMRs activation. However, sTYro3 continued increasing rapidly from the early stage, and that of Mer increased throughout the progression. Roles of Axl may be changed in Mer, because des-γ-carboxy Gas6 levels increasing with Gas6 in the advanced stage of HCC cannot send a signal to Axl.
文摘PC and PS antigens were measured in 370 cases of 12 different diseases and 14 women at third trimester of pregnancy. The PC level was found significantly raised in coronary heart disease, diabetes, nephrotic syndrome, stroke, aplastic anemia and the third trimester of pregnancy, decreased in chronic hepatitis, acute fulminant hepatitis, cirrhosis and deep vein thrombosis (DVT) and no change in acute hepatitis, primary liver carcinoma, Buerger’s disease and leukemia. Total PS was increased in type Ⅱdiabetes, acute lymphocytic leukemia and DVT, and decreased in Berger’s disease, liver diseases, aplastic anemia and the third trimester of pregnancy. In majority of the cases, the total PS and free PS changed parallelly except in Buerger’s disease, DVT and preeclampsia. No correlation was found between PC and PS, PC and AT-Ⅲ.
文摘To editor:Thrombophilia is a group of blood clotting disorders that increase the risk of venous thromboembolism(VTE),which is primarily caused by genetic mutations in proteins involved in the coagulation cascade.These mutations include Factor V Leiden,prothrombin G20210A,protein S deficiency(PSD),antithrombin III deficiency,and methyltetrahydrofolate reductase.Among these,PSD contribute to a significant proportion of thromboembolic events in pregnant women,particularly in those with additional risk factors such as obesity,advanced maternal age,or a history of VTE.1 This correspondence aims to present a series of clinical cases of PSD in pregnant women,highlighting treatment strategies,especially in a hospital in a low-and middle-income country(LMIC)with limited access to advanced therapeutic options.Written informed consent was obtained from all patients involved in this study.We had treated six pregnant women diagnosed with PSD.Among these cases,four had a history of adverse obstetric outcomes,including abortion,preterm birth,or intrauterine fetal death,while the other two cases had recurrent miscarriages(Supplementary Table 1,http://links.lww.com/MFM/A72).These women were identified after the diagnosis of PSD,and each case was carefully monitored throughout pregnancy.
基金Supported by the Excellent Medical Talents Training Program of the First Affiliated Hospital of Guangxi Medical University,Difficult and Critical illness Center,Pediatric Clinical Medical Research Center of Guangxi,No.Gui Ke AD22035219the Key Laboratory of Children’s Disease Research in Guangxi’s Colleges and Universities,Education Department of Guangxi Zhuang Autonomous Region.
文摘BACKGROUND Upper gastrointestinal hemorrhage is a life-threatening manifestation of cow’s milk protein allergy(CMPA).We analyze the clinical characteristics of a case of milk protein allergy manifested as severe upper gastrointestinal hemorrhage.CASE SUMMARY The hospital admitted a 2-month-old male infant due to“melena for 6 days,he-matemesis twice”.The main symptom was melena,initially occurring once or twice per day,then gradually increasing to five or six times per day at their peak.During the course of the illness,the infant vomited blood,but there were no re-ports of vomiting,fever,pale complexion,dyspnea,wheezing,or difficulty brea-thing.Laboratory tests showed hemoglobin level of 87 g/L,platelet count of 349×109/L,and eosinophil percentage of 0.031.Coagulation studies were normal.After avoiding certain foods and feeding with an amino acid formula for 2 weeks,a repeat gastroscopy revealed less bleeding.After six weeks,a positive oral food challenge test confirmed a severe CMPA.At the 4-month follow-up,there was no gastrointestinal bleeding,and the infant was growing and developing well.CONCLUSION The manifestations of milk protein allergy are diverse and nonspecific,with gas-trointestinal bleeding being less common,especially in infants.When infants present with unexplained massive hematemesis,it’s critical to investigate the possibility of CMPA.
文摘Objective: To analyze the dynamic change of serum protein S100b in patients with traumatic brain injury and its clinical value in assessing brain damage. Methods: According to Glasgow coma scale (GCS), 102 cases of traumatic brain injury were divided into mild brain injury group (GCS≥13, n=31, Group A), moderate brain injury group (8<GCS<13, n=37, Group B) and severe brain injury group (GCS≤8, n=34, Group C). Serial S100b concentrations were analyzed by enzyme-linked immunosorbent assay (ELISA) in blood samples taken on admission, 12 h, 24 h, 48 h, 72 h and 7 days after traumatic brain injury. Results: The severe brain injury group showed significantly higher concentration of serum S100b, with earlier increase and longer duration, than the mild and moderate brain injury groups. The patients with higher S100b exhibited lower GCS scores and poor clinical prognosis. The increase in S100b could emerge before clinical image evidence indicated so. Conclusions: Serum S100b can be used as a sensitive index for assessment and prediction of traumatic brain injury severity and prognosis.
基金Supported by National Natural Science Foundation of China （No. 30873399） Doctoral Funds from Ministry of Education （No. 20070162002） and Excellent Talents in Universities Support Plan of Liaoning Province
文摘Objective： To study the effects of Yiqi Huoxue Recipe （益气活血方） and Coxsackie B virus type 3 （CVB3） on the expression of ribosomal protein S20 in rat cardiac myocytes, to explore the pathogenesis of myocarditis induced by CVB3 and the mechanism of Yiqi Huoxue Recipe on gene level, and to further investigate whether Yiqi Huoxue Recipe is an effective prescription for CVB3 myocarditis. Methods： A modified suppression subtractive hybridization （SSH） was used to isolate differentially expressed genes between the CVB3 infection group and the treatment group with Yiqi Huoxue Recipe. The results were further verified by fluorescence RT-PCR. Results： The results of SSH showed that the gene expression of ribosomal protein S20 in the treatment group was higher than that in the CVB3 infection group （P〈0.05）, which agreed with the results of fluorescent RT-PCR. Conclusion： Down-regulation of ribosomal protein S20 mRNA expression might be one of the mechanisms in CVB3 myocarditis, and Yiqi Huoxue Recipe could achieve the treatment of viral myocarditis by regulating the expression of ribosomal protein S20.
文摘Ribosomal protein S 13 gene has been cloned and analyzed in many organisms,but there are few documents relating to insects. In this communication, the full-length cDNA sequence of ribosomal protein S 13 gene in the diamondback moth, Plutella xylostella(Lepidoptera: Plutellidae), was determined by using PCR amplification technique. The features of the ribosomal protein S 13 gene sequence were analyzed and the deduced amino acids sequence was compared with those from other insects. The results of multi-alignment of the amino acid sequences between the diamondback moth and other insect species revealed that this gene sequence is highly conserved in insects. Based on maximum likelihood method, a phylogenetic tree was constructed from 10 different species using PHYLIP software. It showed that nematode is one separate lineage and the five insect speciesbe long to another lineage, whereas those species higher than insects form the third one. The pattern of this phylogenetic tree evidently represented the evolution of different species.
文摘Objective To investigate the mechanism of anticoagulation protein defect in the pathogenesis of unexplained recurrent miscarriage. Methods Fifty-seven patients with a history of unexplained abortion were enrolled as the investigation group for tests of protein C, protein S, antithrombinⅢ(AT-Ⅲ), as well as activated protein C resistance (APC-R). The control group con-sisted of fifty healthy women with a history of normal pregnancy and delivery. Blood samples were obtained for measuring serum activity of protein C, protein S, AT-Ⅲ, and APC-R. Patients with positive APC-R were tested for factorⅤ(FⅤ) Lei-den gene mutation by PCR-RFLP method. Results Of the 57 patients, 12 (21.1%), 1 (1.8%), and 5 (8.8%) cases were found with protein S, protein C, and AT-Ⅲdeficiency respectively, and 13 (22.8%) cases with positive results of APC-R. Of the control group, no protein C or AT-Ⅲdeficiency was ever found, whereas 2 (4.0%) volunteers were presented with protein S deficiency and 3 (6.0%) with positive results of APC-R. No FⅤLeiden gene mutation was identified in all the patients with positive APC-R results. Late spontan-eous abortion cases had higher incidence of anticoagulation protein defect than the early cases. Conclusion Anticoagulation protein defect may play a role in the pathogenesis of fetal loss, especially for those occurr-ing in late stage of pregnancy.
文摘Previous results in this laboratory indicated that in ribosomal protein S12 strepto-mycin-dependent mutants of Bacillus subtilis, the burst size of bacteriophage φ105 was decreased, and the protein synthesis was inhibited, while the DNA and
基金Supported by the Fundamental Research Funds for Rubber Research Institute, CATAS (1630022011014)Key Science and Technology Project of Hainan Province (90107)+1 种基金Basic Scientific Research Operational Fund for Central-level Public-interest Research Institutes (YWFZX2010-9)Special Fund for Science and Technology Research of Public Welfare Trades ( nyhyzx07-033-1)~~
文摘[Objective] ＂Tapping panel dryness （TPD）＂, a syndrome known as tapping incision blocked partly or entirely during latex exploiting, has become the most important factor causing great losses for rubber production. Aiming to elucidate the molecular mechanism of tapping panel dryness occurrence, this study carried out molecular cloning and bioinformatical analysis of a mRPL21 cDNA sequence, a gene associated with TPD. [Method] In a preliminary study, an expressed sequence tag （EST） encoding a deduced protein homologous to mitochondrial 50S ribosomal protein L21 （mRPL21）, which showed to be down-regulated in the latex of TPD-affected rubber trees, was isolated by suppression subtractive hybridization （SSH）. After ESTs assembling and RT-PCR validation, an 853 bp cDNA sequence with an open reading frame （ORF） was cloned, which was named as HbmRPL21 under GenBank accession number of HM230670. [Result] Bioinformatical analysis suggests that HbmRPL21 encodes a deduced polypeptide of 271 amino acids with a theoretical molecular weight （Mw） of 30.52 kDa and isolectric point （pI） of 8.40, and HbmRPL21 is a mitochondrion-targeted protein with a conserved domain of Ribosomal_L21p involving translation. Homology analysis reveals high amino acid sequence identity of mRPL21 from plants, while diversity of that between plant and animal kingdom. [Conclusion] This study laid the basis for further revealing the biological functions of mRPL21 in TPD-affected rubber trees.
文摘BACKGROUND Tacrolimus(FK506)is a key calcineurin inhibitor used to prevent organ transplant rejection and is effective in improving graft survival.However,it is linked to hyperglycemia and insulin resistance,contributing to new-onset diabetes after transplantation and negatively affecting islet function.AIM To study the effects of tacrolimus on the insulin signaling pathway of hepatocytes.METHODS HL7702 cells were treated with different concentrations of tacrolimus(0.1 mg/L,1 mg/L,5 mg/L)for 24 hours.The proteins involved in insulin signaling were detected by Western blotting.RESULTS Compared with the control group,phosphorylation of insulin receptor substrate(IRS)1 at Ser 307 and Ser 323 were increased significantly when the tacrolimus concentration reached 1 and 5 mg/L.Phosphorylation of IRS1 at Ser 1101 was also increased,although not significantly.However,phosphorylation of Ribosomal protein S6 kinase beta-1 at Thr 389 was decreased significantly.The levels of phosphorylated glycogen synthase kinase 3αSer 21 and Ser 9 were increased.Surprisingly,phosphorylation of glycogen synthase at Ser 641 was increased.There was no significant change in the activity of glycogen phosphorylase.CONCLUSION Tacrolimus has no direct effect on hepatic glucose metabolism,but inhibits IRS1-mediated insulin signaling.This may be one of the underlying mechanisms by which tacrolimus induces insulin resistance.
基金Supported by the National Natural Science Foundation of China, No. C03011402, No. C30070690 the Science and Technique Foundation of PLA during the 9th Five-year Plan period, No. 98D063the Launching Foundation for Students Studying Abroad of PLA, No. 98H038the Youth Science and Technique Foundation of PLA during the 10th Five-year plan period, No. 01Q138the Science and Technique Foundation of PLA during the 10th Five-year Plan period, No. 01MB135
文摘AIM: To investigate the transactivating effect of complete S protein of hepatitis B virus (HBV) and to construct a subtractive cDNA library of genes transactivated by complete S protein of HBV by suppression subtractive hybridization (SSH) technique and to clone genes associated with its transactivation activity, and to pave the way for elucidating the pathogenesis of hepatitis B virus infection. METHODS: pcDNA3.1(-)-complete S containing full-length HBV S gene was constructed by insertion of HBV complete S gene into BamH I/Kpn I sites. HepG2 cells were cotransfected with pcDNA3.1(-)-complete S and pSV-lacZ. After 48 h, cells were collected and detected for the expression of β-galactosidase (β-gal). Suppression subtractive hybridization and bioinformatics techniques were used. The mRNA of HepG2 cells transfected with pcDNA3.Incomplete S and pcDNA3.1(-) empty vector was isolated, and detected for the expression of complete S protein by reverse transcription polymerase chain reaction (RT-PCR) method, and cDNA was synthesized. After digestion with restriction enzyme RsaI, cDNA fragments were obtained. Tester cDNA was then divided into two groups and ligated to the specific adaptors 1 and 2, respectively. After tester cDNA had been hybridized with driver cDNA twice and underwent nested PCR twice, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. Amplification of the library was carried out within E. coli strain DH5α. The cDNA was sequenced and analyzed in GenBank with BLAST search after polymerase chain reaction (PCR) amplification. RESULTS: The complete S mRNA could be detected by RT-PCR in HepG2 cells transfected with the pcDNA3.1(-)-complete S. The activity of β-gal in HepG2 cells transfected with the pcDNA3.1(-)-complete s was 6.9 times higher than that of control plasmid. The subtractive library of genes transactivated by HBV complete S protein was constructed successfully. The amplified library contains 86 positive clones. Colony PCR showed that 86 clones contained DNA inserts of 200-1 000 bp, respectively. Sequence analysis was performed in 35 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 33 coding sequences were obtained. These cDNA sequences might be target genes transactivated by complete S protein of HBV. Moreover, two unknown genes were discovered, full length coding sequences were obtained by bioinformatics techniques, one of them was named complete S transactivated protein 1 (CSTP1) and registered in GenBank (AY553877). CONCLUSION: The complete S gene of HBV has a transactivating effect on SV40 early promoter. A subtractive cDNA library of genes transactivated by HBV complete S protein using SSH technique has been constructed successfully. The obtained sequences may be target genes transactivated by HBV complete S protein among which some genes coding proteins are involved in cell cycle regulation, metabolism, immunity, signal transduction, cell apoptosis and formation mechanism of hepatic carcinoma.
