Objective To invetigate the application value of antigen‐antibody joint detection in increasing the detection rate of hepatitis C infection in the patients with hemodialysis .Methods The clinical data of 100 cases of hemodialysis were collected .Hep‐atitis C virus(HCV) core antigen ,HCV antibody and HCV‐RNA were detected by adopting ELISA ,Chemiluminescence and RT‐PCR respectively .The positive expression situation of each detection item in the same specimen was performed statistics .The detec‐tion rates of RNA positive specimen were compared among the single antigen detection ,single antibody detection and joint detection schemes ;at the same time the antigen level of antigen positive specimens ant its RNA copy number were performed the correlation analysis .The antibody levels in the positive antibody specimens were compared between the RNA positive group and the RNA neg‐ative group .Results The single antigen or single antibody detection could cause the missed detection of HCV infection in hemodial‐ysis patients .For the specimens of positive RNA ,the joint detection had the highest detection rate (P<0 .05) ,the antigen level in the antigen positive specimens was positively correlated with the RNA copy number in the corresponding specimens (r=0 .85 ,P<0 .05) ,while the antibody level in the antibody positive specimens had no statistical difference between the RNA positive group and the RNA negative group(P>0 .05) .Conclusion The joint detection of HCV antigen antibody can effectively increase the detection rate of hepatitis C infection in hemodialysis patients .At the same time the HCV core antigen detection also can be used as one of monitoring indicators for the HCV re -infection in the patients with antibody positive .

Objective:To establish an efficient method for expression and purification of vMIP-Ⅱ which encoded by a viral gene and ho-mogued human chemokine in active single strand in E.coli cells. Methods: Cloning with bi-enzyme restriction and lysising bacteria with cold osmoic shock were used for expression,MBP affinity chromatography of fusion protein and self-selective of recombinative peptide for final purification. The expressing and purifying products were detected with SDS-PAGE and Western blotting and identified with inhibitory adhesion experiment for its activity.Results: Fusion protein MBP-vMIP was effeciendy expressed in E. coli at secretive type, and self-cleaved to sparate the vMIP-Ⅱ. The final product single strand vMIP-Ⅱ was active for blocking chemokine receptor CCR5.Conclusion:This is an effective method for obtaining viral chemokine vMIP-Ⅱ of recombinant single strand.The recombinant vMIP-Ⅱ may be useful for the study of diseases involving in chemokiine receptors such as HIV infection,rejection of transplantation and chronic inflammation,etc.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Liver ; pathology ; Liver Cirrhosis ; pathology ; Paeonia ; Rats ; Rats, Sprague-Dawley

Objective To understand the differences between sexes in the clinical and pathological features of patients with mesangial proliferative glomerulonephritis(MPGS).MethodsOne hundred and five patients with MPGS admitted to our hospital were retrospectively studied in clinical and pathological aspects.Results( 1 ) The proportion of male patients were 75 of 105 ( 71.43 ％ ) and that of females were 30 of 105 ( 28.57％ ) ; ( 2 ) The average age of the male patients was ( 40.25 ± 15.50 ) and that of the females was (36.23 ± 15.26) in year.There was no significant difference between the two groups( t =1.206,P =0.231 ) ;(3) There was no significant difference in duration of disease,hematuria,edema,hypertension prevalence and mean blood pressure( P ＞ 0.05 ).The proportion of patients with hematuria was 56.19％ (59/105).The males accounted for 69.33％ ( 52/75 ) and the females were 63.33％ ( 19/30 ) in the main clinical manifestations of nephrotic syndrome.There was no significant difference( x2 =0.352,P ＞ 0.05 ) between the proportion of males and females; (4)Males and females groups had no significant difference( P ＞ 0.05 )on levels of urinary protein,serum albumin,immunoglobulin,complement,urea nitrogen and serum creatinine.Complement decreased in 53 cases,accounting for 53％ of all the participants.The proportion of male patients with renal insufficiency was 24.00％ (18/75),and the proportion of females with renal insufficiency was 13.33％ (4/30).There was no significant difference ( x2 =1.472,P ＞ 0.05 )on the percentage of males and females with renal insufficiency.The mean value of urea nitrogen was higher than the normal levels ; (5) The proportion of male cases with different deposition of immune complexes was 93.06％ (67/72),and the proportion in females were 92.86％ (26/28) in the exception of 5 cases ( male 3 and female 2 ) with no glomeruli in immunofluorescence examination.No significant difference was found between the two groups( x2 =0.001,P ＞ 0.05 ) ; ( 6 ) There was no significant gender differences( x2 =1.696,P ＞ 0.05 ) found in risk assessment.ConclusionThe prevalence of MPGS is higher in male patients than in females,the main clinical manifestations of which were nephrotic syndrome.Patients were found to have a higher rate of hematuria,decreased complement C3,and renal dysfunction than the normal levels.There was no significant difference in gender on the clinical and pathological aspects of MPGS.

ObjectiveTo investigate the influence of timing of definitive operation on prognosis of patients with iatrogenic bile duct injury (IBDI). MethodsThe clinical data of 38 patients with IBDI were retrospectively analyzed. Of the 38 patients, the injury happened in 7 patients in our hospital while 31 patients were transferred in from other hospitals. We analyzed the previous operative records, the clinical manifestations of the patients, the radiological images and our operative findings in the definitive operations. According to the timing of the definitive operation, the patients were divided into the intraoperative repair group (n=26), the early repair group (n=15) and the delayed repair group (n=17). ResultsPatients in the intraoperative repair group had the best short-and long-term results, the shortest length of hospital stay and the lowest total cost. The early repair group, though having worse Results than the intraoperative repair group was still better than the delayed repair group. However, early detection of injuries and the ability to carry out timely corrective procedures depended on the availability of surgical expertise in the hospital. ConclusionsThe timing of definitive operation to provide corrective surgery to IBDI was a vitally important factor which affected prognosis.It is best to repair the injury within the same operation. The repair should be carried out by a doctor with ample experience in bile duct surgery.

10.3969/j.issn.1000-3606.2013.06.018

Objective To examine whether tissue inhibitor of metalloproteinases-1 (TIMP-1) is involved in arterial calcification of chronic renal failure (CRF) rats. Methods CRF model was induced in male Wistar rats by garage daily with 2% adenine 250 mg/kg. The calcification of aorta, femoral artery, renal artery and coronary artery was evaluated histomorphometrically by van Kossa-stained sections at 2-, 4-, 6- and 8-week respectively. RT-PCR and Western blot were used to observe the expressive levels of TIMP-1 mRNA and protein. Expressions of TIMP-1, osteopentin (OPN) and core binding factor α1 (Cbfα-1) protein were analyzed by immunhistochemistry. Results Serum urea nitrogen, creatinine, inorganic phosphate, calcium-phosphorus product and intact parathyroid hormone (iPTH) increased significantly in the model animals compared with control group after 2 weeks (P<0.01). Medial calcification was found in above four arteries of model groups after 6 weeks. RT-PCR and Western blot showed that TIMP-1 expression of model group was significantly higher than that of control group (P< 0.05), and obviously elevated in a time-dependent manner. The expression of TIMP-1 and OPN in calcified aortic smooth muscle cells increased obviously (P<0.05), and positive immunostaining of Cbfα-1 was found. The expression of TIMP-1 was positively correlated with OPN and Cbfα-1 (r=0.317, P=0.000; r=0.485, P=0.000). Conclusions The pathology of arterial calcification in CRF rats induced by adenine is similar to CRF patients, which may serve as a useful model of CRF with arterial calcification. The up-regulation of TIMP-1 seems to participate in the formation and development of vascular calcification in CRF.

Objective To explore the repair possibilities of endothelial progenitor cells (EPCs)in peripheral blood in patients with different extents of obstructive sleep apnea (OSA) through measuring the levels of pro-angiogenic factors and different subgroups EPCs in peripheral blood in patients with OSA. Methods Ninety adult patients with OSA, 30 healthy controls with matched age and gender were enrolled for this study. The subjects performed Polysomnography, were divided in-to four group based on Apnea Hypopnea Index (AHI). The serum levels of HIF-1α, SDF-1αand VEGF were assessed by ELISA. Mononuclear cells were isolated from peripheral blood with density gradient centrifugation, and flow cytometry was used to detect levels of CD133+KDR+EPC, CD133+CD34+EPC, CD34+KDR+EPC and ALDHloCD34+KDR+EPC based on AL-DH activity, and CD133, CD34, PE-KDR related cell surface markers. Results The levels of CD133+KDR+EPC, CD133+CD34+EPC, CD34+KDR+EPC were higher in OSA groups than those of control group, both of which were higher in severe OSA group than those of in mild and moderate OSA groups. The levels of ALDHloCD34+KDR+EPC were higher in mild and moderate OSA groups than that of the control groups, and the levels of ALDHloCD34+KDR+EPC were significantly lower in se-vere OSA group than those of control, mild and moderate OSA groups. Serum levels of HIF-1α. VEGF were significantly high-er in OSA groups compared to those in control groups, both of which were higher in severe OSA group than those of mild and moderate OSA groups. Serum levels of SDF-1αwere significantly lower in severe OSA groups than those of mild, moderate OSA and control groups (P<0.05). Conclusion The mobilization and recruitment of different subtypes of EPCs are obvious-ly increased in patients with OSA, but ALDHloCD34+KDR+EPC with vascular repair capacity keeps to invariability, even de-creases in patients with severe OSA, which results in endothelial damage, and increases the risk of cardiovascular disease.

Objective To investigate the related factors of early liver cancer recurrence associated with elevated fasting glucose levels after radical resection of liver cancer.Methods The retrospective cohort study was adopted.The clinical data of 145 patients with liver cancer who were admitted to the Nanfang Hospital of Southern Medical University from October 2009 to June 2013 werc collected.After radical resection of liver cancer, 111 patients with average level of fasting blood glucose (FBG) ＜ 6.1 mmol/L and 34 patients with level of FBG ≥ 6.1 mmol/L were divided into the normal group and the hyperglycemic group, respectively.The blood glucose levels of patients during the period of hospital stay and follow-up were observed.The status of early recurrence and risk factors affecting recurrence in the 2 groups were analyzed.Follow-up of outpatient examination was applied to patients up to July 1, 2015 with the first recurrence as the end point.Patients had reexaminations once every month within postoperative month 6 and once every 3 months after 6 months.The general information [gender, age, FBG, underlying hepatopathy, preoperative AFP, alanine transaminase (ALT), aspartate aminotransferase (AST), Child-Pugh stage, Barcelona Clinic Liver Cancer (BCLC) stage, history of alcohol drinking], surgical data (surgical method, operation time, number of blood transfusion, volume of intraoperative blood loss), pathological data (histopathological differentiation, number of tumor nodules, diameter of maximal tumor, liver cirrhosis) and follow-up data (postoperative AFP, imaging findings, recurrence time, preventive chemotherapy) were collected.Measurement data with normal distribution were presented as-x ± s and analyzed by the t test.Measurement data with skewed distribution were presented as M (range) and analyzed by the non-parametric test.Count data were analyzed using the chi-square test.With the first recurrence as the end point, the tumor-free survival rate was drawn using the Kaplan-Meier method, and the comparison was analyzed by the logrank test.The risk factors affecting recurrence were analyzed using the Kaplan-Meier method for proportional hazards assumption test firstly, and then eligible factors were done using Log-rank test in the univariate analysis.The multivariate analysis was done using the COX model.Results The postoperative 1-, 2-year overall recurrence rates of liver cancer were 28.3％ (41/145) and 45.5％ (66/145).And 1-, 2-year recurrence rates of liver cancer were 21.6％ (24/111) and 36.9％ (41/111) in the normal group and 50.0％ (17/34) and 73.5％ (25/34) in the hyperglycemic group, respectively, showing significant differences between the 2 groups (x2=10.335, 14.053, P ＜ 0.05).The univariate analysis showed that FBG, Child-Pugh stage, volume of intraoperative blood transfusion and postoperative AFP were risk factors affecting tumor-free survival rate after radical resection of liver cancer (x2 =17.591,6.492, 10.690,12.820, P ＜ 0.05).The tumor-free survival rates at postoperative month 24 in the normal group and hyperglycemic group were 63.1％ and 26.5％ respectively, showing significant difference between the 2 groups (x2=17.591, P ＜ 0.05).The results of multivariate analysis showed that level of FBG ≥6.1 mmol/L, volume of intraoperative blood transfusion ＞ 200 mL and postoperative level of AFP ＞ 8.1 pg/L were independent risk factors affecting tumor-free survival rate after radical resection of liver cancer (RR =2.542, 2.028, 2.724, 95 ％ condifence interval : 1.529-4.225,1.183-3.479,1.635-4.538, P ＜ 0.05).Conclusions Elevated FBG level has a stimulative effect on early recurrence of tumor after radical resection of liver cancer.As a result, monitoring and controlling of blood glucose after operation is helpful in decreasing the early recurrence rate of patients with liver cancer.

Objective To explore the role of microRNA (miR)-22 and miR-1825 in the diagnosis and differential diagnosis of juvenile systemic lupus erythematous (JSLE).Methods The cases of JSLE hospitalized in Capital Institute of Pediatrics Teaching Hospital Affiliated to Peking University from June 2013 to May 2014 were selected as study group.The cases with systemic juvenile idiopathic arthritis (sJIA),nephrotic syndrome (NS),Kawasaki disease (KD),Henoch-Schonlein purpura(HSP) were selected as patients control group.The healthy children were selected as healthy control group.The expression levels of miR-22 and miR-1825 in the plasma of JSLE,sJIA,NS,KD,HSP and healthy children were detected by using real-time PCR respectively.Receiver operating characteristic curve (ROC) analysis was performed to evaluate the value of miR-22 and miR-1825 miRNA as a biomarker with the sensitivity and specificity.Three data bases,included Targetscan,PicTar and miRanda,were applied to predict the target gene.The target gene was analyzed by adopting Gene Ontology (GO) in terms of molecular function,biological process and cellular component,and by adopting Kyoto Encyclopedia of Genes and Genomes (KEGG) in terms of pathway.Results Compared with healthy children,the amount of miR-22 and miR-1825 in JSLE patients were lower,and there were significant differences(t =-3.076,-9.054,P ＜0.01,0.000 1).The levels of the miR-22 and miR-1825 miRNAs in controls of sJIA,NS,KD,HSP were significantly higher than those of JSLE (t =-4.410,-4.477,-4.494,-2.971,all P ＜ 0.000 1;t =-9.043,-6.045,-10.416,-8.712,all P ＜ 0.000 1),but there was no difference compared with healthy children(all P ＞ 0.05).The area under ROC curve(AUC) of miR-22 between JSLE and healthy children was 0.777.The AUC of miR-1825 between JSLE and healthy children was 1.000.The AUCs between JSLE and controls of sJIA,NS,KD,HSP of miR-22 were 0.731-1.000.The AUCs between JSLE and controls of sJIA,NS,KD,HSP of miR-1825 were 0.939-1.000.There was positive relation between the amount of miR-22 and complement C3 in plasma(r =0.493,P =0.027).Conclusions The amount of miR-22 and miR-1825 in the plasma of JSLE embrace the potential of distinguishing JSLE from healthy children,sJIA,NS,KD,HSP.MiR-22 has the ability to predict the activity of JSLE.