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Vav-family proteins are guanosine nucleotide exchange factors(GEFs)and one of RhoGEFs family numbers.Vav proteins can influence cell signaling transduction, cytoskeleton recombination and adherence migration in special cells by RhoA, RhoG and Rac-1 pathway.There are 80 kinds of RhoGEFs in mammalian cell.Vav proteins play an important role in hematological system, nervous system, cardiovascular system and immune system.So the function of vav protein in immune system was reviewed in this paper.
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Vav-family proteins are guanosine nucleotide exchange factors (GEFs) and one of RhoGEFs family numbers.Vav proteins can influence cell signaling transduction,cytoskeleton recombination and adherence migration in special cells by RhoA,RhoG and Rac-1 pathway.There are 80 kinds of RhoGEFs in mammalian cell.Vav proteins play an important role in hematological system,nervous system,cardiovascular system and immune system.So the function of vav protein in immune system was reviewed in this paper.
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Objective@#To investigate the role of midkine(MK)in the pathogenesis of Henoch-Schonlein purpura(HSP) and Henoch-Schonlein purpura nephritis(HSPN).@*Methods@#In the case group, 35 cases were hospitalized in the pediatric kidney rheumatism immunology ward of Shengjing hospital affiliated to China Medical University from December 2016 to January 2018.Among them, 10 cases were HSP, 25 were HSPN.According to quantitative level of 24-hour urine protein, HSPN group was divided into HSPN(nephrotic level of proteinuria)group of 15 cases and HSPN(non-nephrotic level of proteinuria)group of 10 cases.The control group consisted of 12 healthy cases who underwent physical examination at outpatient department in the same period in the developmental pediatric clinic of our hospital.Blood samples were collected to detect MK.The other clinical datas including renal function, 24-hour urine protein quantitative, immunoglobulin, etc were collected.The serum MK and renal function indexes were compared among groups.The correlation between MK and various clinical indicators was analyzed, and the receiver operating characteristic(ROC) curve was used to analyze the diagnostic significance of MK for HSP and HSPN.@*Results@#MK level of case group was higher than that of healthy control group[(289.34±160.70)pg/ml vs.(100.03±56.75)pg/ml, P<0.05]. Moreover, the difference of MK concentration among the HSPN(nephrotic proteinuria)group, the HSPN(non-nephrotic proteinuria)group and the HSP group was still statistically significant[(449.91±141.91)pg/ml vs.(244.04±89.15)pg/ml vs.(175.94±46.30)pg/ml, P<0.05]. MK was positively correlated with urine microalbumin(r=0.54), IgA(r=0.132), IgE(r=0.304), urine β2 microglobulin(r=0.483), 24-hour urine protein /body weight(r=0.503), and urine transferrin level(r=0.509)in the case group(P<0.05). According to the ROC curve, the area under ROC of MK for predicting the diagnosis of HSP was 0.908(95%CI 0.828-0.988). The optimal value in predicting the diagnosis of HSP was 182.762 pg/ml, with sensitivity and specificity of 81.4% and 91.7%.The area under ROC of MK in predicting HSPN was 0.947(95%CI 0.888-1.000), and the optimal value of predicting HSPN was 218.186 pg/ml, with sensitivity and specificity of 84.0% and 95.5%.@*Conclusion@#MK may be involved in the pathogenesis of HSP and HSPN.It can provide the basis for clinical diagnosis of HSP and HSPN, and has significance in evaluating the degree of renal damage of HSPN.
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Objective To investigate SCCmec types in clinical isolates of methicillin-resistant Staphylococcus epidermidis (MRSE) carrying psm-mec.Methods We collected 165 strains of Staphylococcus epidermidis identified by automated microbiological identification system and screened MRSE by PCR amplification of esp and mecA gene.Strains with psm-mec were identified by amplification of psm-mec,fudoh and p221 DNA fragment;mec,ccr and SCCmec typing was conducted by multiplex PCR assay.Results Among 138 strains of MRSE,29 strains were identified as MRSE with psm-mec,and the carrying rate was 17.58%.Results of mec and ccr typing by multiple PCR showed that MRSE with psm-mec carried Class A mec,but the ccr type had obvious diversity.Results of SCCmec typing showed that all strains with psm-mec belonged to type Ⅱ and/or Ⅲ SCCmec.Conclusion Clinical isolates of MRSE with psm-mec carry homologous type Ⅱ and/or Ⅲ SCCmec harboring Class A mec.
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Objective To discuss the application value of modified Hodge test(MHT) for screening carbapenemase-producing Enterobacteriaceae.Methods The 24 Enterobacteriaceae reduced susceptibility to carbapenems were detected by MHT.At the same time,polymerase chain reaction(PCR) was used to detect carbapenemase genes of KPC,NDM,IMP,SIM and VIM.PCR products were sequenced and the results were compared with the sequences of Gen Bank database.Comprehensive analysis the application value of MHT and PCR to detect carbapenemase.Results Among these 24 strains,13 stains appeared to produce carbapenemase by MHT,5 positive strains were found to carry carbapenemase genes by PCR.By comparing with the sequences of Gen Bank database 1 strain were confirmed to KPC-2 and 4 strains were confirmed to IMP-4.We found that 4 strains of Enterobacteriaceae,detected carbapenemase by MHT and PCR at the same time.9 strains of MHT were positive,but we couldn′t detect the carbapenemase genes.1 strain of MHT was negative,but carbapenemase gene was found in the strain.Conclusion The value of MHT to screen carbapenemase-producing Enterobacteriaceae is necessary to further study.
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Objective To investigate the SCCmec types of clinically isolated methicillin‐resistant Staphylococcus epidermidis (M RSE) .Methods Eighty‐four strains of clinically isolated Staphylococcus epidermidis identified by the fully automatic microbio‐logical identification system were collected and performed the MRSE identification by PCR for amplifying esp and mecA genes and SCCmec typing .Its distribution characteristics were analyzed .Results Esp gene was amplified in 84 strains and the detection rate of mecA was 76 .19% (64/84) ,in which the MRSE detection rates in blood ,sputum ,urine and wound secretion were 76 .8% , 68 .8% ,100% and 71 .4% respectively .The multiple PCR amplification displayed that among 64 strains of MRSE ,19 strains were SCCmec simple type ,in which 19 strains were SCCmec type Ⅰ and 3 strains were SCCmec type Ⅲ ;42 strains were SCCmec mixed type ,in which 2 strains were SCCmec mixed type Ⅰ and Ⅱ ,14 strains were SCCmec mixed type Ⅰ and Ⅲ ,12 strains were SCCmec mixed type Ⅰ ,Ⅱ and Ⅲ ,5 strains were SCCmec mixed type Ⅱ and Ⅲ ,a strains were and SCCmec mixed type Ⅲ and Ⅳ .Conclu‐sion The SCCmec type in clinically isolated MRSE shows obvious diversity and its majority is SCCmec mixed type .
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Objective To investigate the genetic location of SCCmec-associated psm-mec in Staphylococcus hominis isolated from blood culture,and to lay a foundation for further functional studies of psm-mec in Staphylococcus hominis.Methods 25 strains of Staphylococcus hominis isolated from positive blood culture were collected.mecA and psm-mec gene were amplified by PCR,and the SCCmec types were determined by the results of multiplex PCR assay.For analyzing the genetic location characteristic of psm-mec in SCCmec,three pair special PCR primers were used to measure mecR1/psm-mec,psm-mec/xylR and fudoh respectively.Results There were 21 strains of methicillin-resistant Staphylococcus hominis and 4 strains of methicillin-sensitive Staphylococcus hominis. The positive rate of psm-mec gene in methicillin-resistant Staphylococcus hominis was 47.6%,and no psm-mec gene was found in methicillin-sensitive Staphylococcus hominis.Among psm-mec positive strains,2 strains belonged to SCCmecⅢ,5 strains belonged to SCCmecⅢ-like,and 3 strains belonged to new SCCmec types.All of the 10 psm-mec positive strains were mecR1/psm-mec,psm-mec/xylR and fudoh gene positive.Conclusion SCCmec-associated psm-mec extensively exists in methicillin-resistant Staphylococ-cus hominis isolated from positive blood culture,which distributes mainly in typical SCCmecⅢ,SCCmecⅢ-like and new SCCmec types and locates between mecR1 and xylR gene.
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Objective To investigate the relationship between suspected ankylosing spondylitis and HLA‐B27 antigen by detec‐ting the positive frequency of HLA‐B27antigen in 872 suspected AS patients ,and evaluate its clinical significance .Methods The positive frequency of HLA‐B27 on the T lymphocyte membrane were detected by flow eytometer in 872 suspected AS patients .Re‐sults Among the 872 suspected AS patients the ratio between male and female was 1 .8∶1 ,the positive rate of antigen HLA‐B27 was 27 .29% ,and the male and female patients′positive rates of HLA‐B27 antigen were 32 .50% and 17 .95% ,respectively (P<0 .05) .The male and female patients′ expression percentage of B27+ /B7- monoclonal antibody were 39 .16 ± 42 .79 and 20 .96 ± 33 .86 ,respectively(P<0 .05) .The male and female patients′mean fluorescence intension of B27+ /B7- monoclonal antibody were 5 .35 ± 5 .44 and 3 .35 ± 3 .87 ,respectively(P<0 .05) .Conclusion The patients with AS are strongly associated with HLA‐B27 an‐tigen .Detection of HLA‐B27 antigen expression intensity in suspected AS patients with FCM is helpful to diagnosis and differential diagnosis of AS .
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Objective To evaluate the diagnostic value of procalcitonin(PCT),C-reactive protein(CRP),prealbumin(PA)and white blood cell (WBC)count measurements in patients with severe pneumonia.Methods The serum samples of 34 patients with severe pneumonia,68 non-severe pneumonia patients and 40 healthy volunteers were collected.Serum concentrations of PCT,CRP, PA and WBC count of all samples were determined.Results The levels of PCT,CRP,PA and WBC in patients with severe pneu-monia were (24.07±34.77)ng/mL,(98.75 ±69.63)mg/L,(105.65 ±68.88)mg/L,(12.64±7.62)×109/L,which were signifi-cantly higher than those in non-severe pneumonia patients and healthy group(P <0.05).According to ROC analysis,the sensitivity, specificity and Youden index of PCT were 64.7%,77.9%,0.426.Conclusion The level of serum PCT could be used as good bio-marker for severe pneumonia.Detection of PCT,CRP and WBC together plays an important role in the diagnosis of severe pneumo-nia.
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Objective To construct mutant strains of methicillin resistant Staphylococcus epidermi-dis (MRSE) with psm-mec gene deletion and to investigate the function of psm-mec gene.Methods The drug sensitivity test and DNA sequence analysis were performed to screen out the tetracycline and chloram -phenicol sensitive clinical strains of MRSE , whose upstream and downstream sequences of psm-mec gene were identical to those of the Staphylococcus epidermidis reference strain RP62A.The recombinant plasmid pBT2-Δpsm-mec was constructed by using the fusion PCR and a temperature sensitive shuttle plasmid .After being identified , the plasmid was transformed into the Staphylococcus aureus RN4220 strain by electropora-tion, and then transformed into the selected clinical isolates of MRSE .The mutant strains of MRSE with psm-mec deletion were screened out and identified after homologous recombination .The differences in biofilm formation between the mutant and wild-type strains were analyzed for further elucidation the relationships be-tween the psm-mec gene and biofilm formation in MRSE strains .Results Three clinical MRSE isolates for the construction of mutant strains with psm-mec gene deletion were screened out and identified by using drug sensitivity test and sequence alignment analysis .The mutants constructed via homogenous recombination were screened out and identified .Compared with the corresponding wild-type strains, the three mutants with psm-mec gene deletion showed significantly decreased ability of biofilm formation , demonstrating that the psm-mec genes strains induced the biofilm formation of MRSE .Conclusion The Δpsm-mec mutant strains were successfully constructed .The psm-mec gene played an important role in the biofilm formation of Staphy-lococcus epidermdis.
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Objective To investigate the mechanism of one carbapenems resistant Raoultella planticola( R.planticola) isolate.Methods This is an experimental study.R.planticola was isolated from a patient′s drainage fluid from orthopedic department in November 2010 in Sichuan Provincial People′s Hospital.Minimum inhibitory concentration of R.planticola to 13 antibiotics was determined by using the agar dilution method.Modified Hodge test was used to detect carbapenemase .EDTA synergistic test was performed to research metallo-beta-lactamase.The genes coded the β-lactamase were amplified by polymerase chain reaction ( PCR ) , including class A carbapenemase ( KPC ) , class B carbapenemases (NDM, IMP, VIM, SIM), extended spectrum beta-lactamases[ESBL(CTX, TEM, SHV)], and AmpCβ-lactamases ( FOX, EBC, ACC, DHA, CIT, MOX).Results The susceptibility test showed that R.planticola was resistant to 9 antibiotics.MIC value of meropenem for R.planticola was up to 32 mg/L.R.planticola kept intermediary to imipenem , whereas it was susceptible to cefepime , amikacin and polymyxin B.Modified Hodge test and EDTA synergistic test were positive in R.planticola.Class B carbapenemase (IMP) gene and two extended spectrum β-lactamases(CTX, SHV) genes were positive by PCR.The genes were conformed as IMP-4, CTX-M3 and SHV-12 by sequencing and compared with GenBank.Other resistant genes were negative.Conclusion IMP-4 was identified in R.planticola, the combined produce IMP-4 and ESBLs might be the main mechanism of R.planticola resistant to carbapenems.
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Objective To explore the establishment of peptide mapping database of Candida albicans ,laying the foundation for rapid diagnosis of Candida albicans infection .Methods 96 Candida albicans were collected clinically ,and its DNA was extracted . Polymerase chain reaction(PCR) was used to amplify the ITS1-5 .8S-ITS2 gene fragments and restriction endonucleases were a-dopted to identify them .Surface enhanced laser desorption ionization-time of flight-mass spectrometry(SELDI-TOF-MS) instrument was applied to detect the Candida albicans peptide mapping ,and Ciphergen ProteinChip software was used to collect data automati-cally .The established peptide mapping database was verified by confirmed Candida .Results According to restriction fragment length polymorphism analysis ,96 strains were confirmed as Candida albicans .15 peptide peaks were captured by SELDI-TOF-MS chips .Five peptide peaks of them with stable expression were screened out ,and the similarity analysis software was used to estab-lish peptide mapping database of Candida albicans .More than 95% of similarity was found between peptide mapping of Candida albicans and established database ,while less than 50% was found between peptide mapping of other Candida species and database . Conclusion The establishment of peptide mapping database of Candida albicans provides a theoretical basis for the rapid diagnosis of Candida albicans infection .
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Objective To establish protein fingerprints of common bacteria in clinics and to lay a foundation for rapid identification of bacteria.Methods Strains of Escherichia coli,Klebsiella pneumoniae,Pseudomonas aeruginosa and Staphylococcus aureus were detected by surface enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF MS).Stable expression protein peaks were screened and the data was input into the self-constructed Fingerwave software for identification of target bacteria by protein fingerprint comparison.Two hundred and fifty-six clinical isolates,including E.coli,K.pneumoniae,P.aeruginosa and S.aureus were detected and the data was compared with constructed database to evaluate its diagnostic value.Results The protein fingerprints including four common bacteia was used to identify the target bacteria with identification rate of 93.1％ (54/58) for E.coli,87.2％ (75/86) for K.pneumoniae,96.2％ (60/63) for P.aeruginosa and 96.2％ (51/53) for S.aureus,respectively.Conclusion Common bacteria can be rapidly identified by using the protein fingerprint comparison,which provides a powerful tool for bacterial identification.
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Help T cell 1/Help T cell 2 and some cytokines disequilibrium can give a suitable explanation for hypersensitivity and hypogammaglobulinemia in primary nephritic syndrome(PNS) patients. The disturbance of regulatory T cell(Treg cell) and Th17 cell can lead to correlated cytokines derangement, which explained the pathogenesis of PNS from another aspect. Refractory nephrotic syndrome can be effectively treated by rituximab followed the percentage of regulatory T cell increasing, which indicated that Treg may play an important role in pathogenesis of PNS.
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Objective To establish protein fingerprinting identification model of Pseudomonas aeruginosa (P. aeruginosa) and to lay a foundation for rapid identification of P. aeruginosa by proteinchip golden array. Methods Sixty-four P. aeruginosa and one hundred and ninety-nine control bacteria identified in our laboratory were collected and divided into training and testing group. Surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS) and proteinchip golden array were used to detect the protein profiling of the bacteria. Data were automatically collected by Ciphergen Proteinchip Software and protein markers of P. aeruginosa were screened by BioMarker Wizard Software. Classification tree model was developed and validated by BioMarker Patterns Software. The model was blindly tested with twenty-nine P. aeruginosa and sixty-four control bacteria. Results Eighty protein peaks were detected between 3000 and 20 000, among which fifty-eight ones showed significantly difference between P. aeruginosa and the control bacteria (P＜0.01). By BioMarker Patterns Software, one protein peak ( M/Z at 14 045.2) was chosen to develop a classification tree model. The results exhibited with sensitivity of 96. 55% and specificity of 100%. Conclusion Proteinchip golden array has the potential for rapid identification of P. aeruginosa.
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Objective To search for protein markers in urine from patients with diabetic nephropathy by proteomic method and discuss its clinical significance in laboratory diagnosis of diabetic nephropathy. Methods This study included 129 patients with diabetic nephropathy, 61 diabetes mellitus patients, and 102 healthy volunteers. The urinary protein profiles were obtained using surface-enhanced laser desorption-ionization time of flight mass spectrometry (SELDI-TOF-MS) and Au Chip (ProteinChip Gold Array). The differential peaks were screened by Biomaker Wizard software and the decision tree pattern was developed by Biomarker Patterns Software (BPS). The model was blindly tested to validate diagnostic efficiency. Some differentially expressed protein was preliminarily identified according to the molecular weight as compared with mass spectrometry data of standard proteins. Results Totally 40 distinguished protein peaks(t value: - 9.81-24.52, P < 0.05) were obtained after comparing the samples between diabetic nephropathy and the control groups. The peak with m/z 66 916 was automatically screened by BPS to develop decision tree pattern. The pattern was blindly tested and yielded a sensitivity of 98.7% (78/79) and a specificity of 98.2% (111/113). After we compared results from diabetic nephropathy with those from diabetes mellitus, twenty-four differential peaks were obtained in diabetic nephropathy (t value: -6.95-14.45,P < 0.05). The peaks with m/z 4 008, 11 619 and 66 916 were automatically screened by BPS to establish decision tree pattern. The model was blindly tested and yielded the sensitivity(129/129) and specificity(61/61) of 100%. After we compared our results with mass spectrometry data of standard proteins, the four differentially expressed proteins with m/z 11 619, 23 529, 66 916 and 79 378 were supposed to be β_2-microglobulin, α1-microglobulin, albumin and transfcrrin. Conclusion The preliminary results suggest that these SELDI-TOF and Au chip have the potential application value in identification of protein source and early diagnosis of diabetic nephropathy, and evaluation of renal injury.
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Objective To investigate the apoptosis of cultured normal human hepatocyte HL-7702 cells induced by glycodeoxycholate(GCDC)and to explore its possible mechanism.Methods HL-7702 cells were incubated with various concentrations(100,150,200 and 250 ?mol/L)of GCDC.The changes of cellular morphology were observed under optical microscope.The apoptosis rate of HL-7702 was determined by Annexin V-FITC/PI double staining.The changes of HL-7702 cell intracelluar \[Ca2+\]i were determined with Fluo-3/AM load technique.The mRNA expression levels of Bcl-2/Bax in HL-7702 cells were analyzed by RT-PCR.Results Typical apoptotic morphological changes were observed after HL-7702 cells had been treated with 150 ?mol/L GCDC for 24 h;HL-7702 cells could be induced to undergo apoptosis in a concentration-dependent manner after 100,150,200,and 250 ?mol/L GCDC treatment for 24 hours.The apoptosis rates were(13.16?2.9)%,(20.3?3.0)%,(25.02?2.1)% and(45.02?3.5)%,which were markedly higher(P