Objective:To amplify and sequence of the variable region genes of anti CD3 McAb.Methods:The V H?V L genes were amplified by RT PCR from total RNA that were extracted from WuT3 hybridoma.Recombinant cloning vector was constructed and sequenced after the enzyme digestion.Results:It showed that V H gene consisted of 363 bp encoding amine acid residues,belongs to mouse heavy chain subgroup IIB;V L gene consisted of 330 bp encoding amine acid residues,belongs to mouse ? light chain subgroup III.Comparing with Kabat database,the V H?V L genes were in agreement with the characterization of DNA sequences present in the mouse Ig V H?V L regions respectively.Conclusion:The success of cloning of the V H?V L genes of WuT3 McAb lay a good foundation for the construction and expression of chimeric antibody.

BACKGROUND:Pain is the main reason for the recovery of knee function after total knee arthroplasty. OBJECTIVE:To explore and analyze the influencing factors of pain after total knee arthroplasty.METHODS:160 patients with knee osteoarthritis underwent total knee arthroplasty. At 12, 24 and 48 hours after total knee arthroplasty, patients were evaluated with visual analogue scale (VAS) and knee function was assessed with KSS score preoperatively and 90 days postoperatively. Factors, including operation time, age, gender, body mass index, anesthesia method, and preoperative score of KSS were analyzed by Pearson correlation and multiple linear regression. These statistics method also was used to analyze the relationship between the postoperative 90 days score of KSS and postoperative VAS score at each time point. RESULTS AND CONCLUSION:(1) Pearson correlation analysis and multiple linear regression analysis showed that operation time, gender, and body mass index had been correlated positively with VAS score at 12, 24 and 48 hours postoperatively. (2) Patient age had been correlated negatively with VAS score at 12, 24 and 48 hours postoperatively. (3) VAS score at 12, 24 and 48 hours postoperatively had been correlated negatively with KSS score at 90 days postoperatively. (4) Gender, operation time, body mass index and age are al the influencing factors of the pain after total knee arthroplasty. Pain after total knee arthroplasty affects the recovery of knee function.

Objective To study the difference of oxygen free radical metabolism between healthy native Tibetans and mi- grated Hans at different altitude.Methods The activity of Total-antioxidation capability(T-AOC),superoxide dis- mutase(SOD),glutathione peroxidase(GSH-PX)and the content of,reactive oxygen species(ROS),malondialdehyde (MDA)and nitric oxide(NO)in serum in healthy native Tibetans and migrated Hans at different altitude were meas- ured.Results The activity of T-AOC,SOD,GSH-PX and the content of NO were increased in serum in native Tibet- ans group than that in migrated Hans group(P

Objective To identify the outcome of pregnancy and the alteration of renal function in women with nephrotic syndrome. Methods From 2003 to 2007, 59 pregnant women with nephrotic syndrome in our hospital were enrolled in the study. Their clinical data were retrospectively analyzed, including the time of kidney disease onset, 24-hour proteinuria, serum albumin, serum creatinine, blood uric acid, blood pressure, fetal survival, fetal mortality, rate of premature delivery, birth weight of the newborn, and proteinuria, renal function, blood pressure of the patients during their postpartum follow-up. Logistic regression analysis was used to identify the risk factors influencing the outcome of the patients and the newborns. Results The average gestational week was (20.35±9.40) weeks when proteinuria was detected in these pregnant women. The 24-hour proteinuria ranged from 3.5 to 15 g/24 h (median 5.1 g/24 h). The serum albumin was between 10 and 28 g/L (median 22.5 g/L). The serum creatinine was between 32 and 825 μmol/L (median 84 μmol/L) and the serum uric acid ranged from 196 to 793 μmol/L (median 385.5 μmol/L). Pregnancy-induced hypertension syndrome occurred in 75% of the patients, among whom 55.5% suffered from preeclampsia. Forty-three (72.9%) newborns survived , among whom 76.7% (33/43) were premature births and 62.8% (27/43) were low birth weight infants. 50% of the pregnant women still had nephrotic syndrome after delivery. 75% of 24 patients with pre-existing chronic glomerulonephritis had increased proteinuria during pregnancy. Among the 38 patients with renal insufficiency, 36.8% had poorer renal function after delivery. 23.7% of the patients progressed into end stage renal failure after delivery, 80% of whom had serum creatinine ≥ 265 μmol/L. 89% of the patients had persistent hypertension after childbirth. The Logistic regression analysis indicated hyperuricemia during pregnancy (P=0.018, OR=1.012) and the increase of serum creatinine (P=0.039, OR=1.005) were risk factors of renal failure in pregnant women after delivery. Hyperuricemia (P=0.012, OR=1.006)was the risk factor of fetal death. Conclusions Pregnancy with nephrotic syndrome leads to a low fetal survival. Hyperuricemia is the most important risk factor of the poor outcome of pregnant women and newborn.

Objective To compare the efficacy of continuous ambulatory peritoneal dialysis (CAPD) and hemodialysis (HD) on polycystic kidney disease (PKD) patients with end-stage renal disease (ESRD). Methods Retrospective analysis was made on 29 patients with PKD who carded out dialysis therapy for over 3 months in our department from January 2001 to December 2007. They were divided into the CAPD group (10 cases, 34.5%) and HD group (19 cases, 65.5%). Ten cases of non-PKD CAPD patients were randomly selected as the control, who matched the CAPD group in terms of age and gender. The patient information was recorded, such as general data, initial dialysis data, comphcations, survival time, quit of dialysis or death, etc. Kaplan-Meier method and Log-rank test were adopted to analyze the survival rate. Results The survival rates of 1-, 3- and 5-year for the CAPD group were 90%, 75% and 25% respectively, while for the HD group were 94.4%, 67.6%, and 48.3%, and for the control were 83.3%, 44.4% and 22.2% respectively, with no significant differences among 3 groups (P>0.05). group and the control were quite similar. The incidence of peritonitis for the CAPD group (0.62 times/patient year) was similar to that for the control (0.30 times/patient year)(P>0.05). The duration of the lust peritonitis[(23.5±4.0) months vs (20.0±15.8) months] and the catheter exit-site infection (0 time vs 1 time) for two groups were similar as well (P>0.05). One patient had hernia in CAPD group and no patient in control group had hernia. The incidence of peritoneal dialysate leakage was similar between these two groups. In the HD group, two patients (10.5%) had cerebral hemorrhage resulting in death, and 10 patients (52.6%) had cystic hemorrhage, 5 out of whom underwent operation due to repeated cystic hemorrhage and 2 cases received unilateral nephrectomy because of severe hemorrhage. No patient in CAPD group had cerebral hemorrhage but 1 patient (10%) had cystic hemorrhage and recovered after conservative treatment. The hemorrhage complication incidence of CAPD group was significantly lower than that of HD group (P<0.05). Conclusions The prognosis and complication incidence in PKD and non-PKD patients treated with CAPD are similar. The prognosis of PKD patients treated with CAPD or HD is also similar, and the risk of hemorrhage complications of PKD patients treated with CAPD may be decreased compared with those treated with HD. PKD patients can choose HD or PD as the initial therapy of ESRD unless existence of hernia or intolerance. PKD is not the contraindication of PD.

Objective To investigate the clinical characteristics and angiographic findings of cardiogenic shock(CS)following acute myocardial infarction(AMI) in elderly patients.Methods Between January 2015 and April 2016,we carried out a retrospective observational analysis of consecutive elderly patients in Tianjin Chest Hospital,who suffered CS-complicating AMI.Emergency angiography and percutaneous coronary intervention(PCI) were performed after admission.All selected patients were divided into CS and non-CS groups according to whether CS occurred.Electrocardiograph (ECG),cardiac enzyme testing,and ultrasound cardiography were performed after admission to monitor the occurrence of CS.Results The incidence of CS-complicating AMI was 8.33％ (34/408) in elderly patients.Among all CS patients enrolled,the aged patients accounted for 91.89 ％ (3 4/3 7).In-hospital mortality rate was 2 9.41 ％ (10/3 4).There were significant differences between two groups in WBC,H s-CRP,blood glucose,CR and ALT (t =2.403,4.596,6.778,6.109,each P＜0.05).The NT-Pro BNP level,the time of FMC,the frequency of left main and multivessel disease were higher in the CS group than in the non-CS group (each P ＜ 0.05).Conclusions Elderly patients are bearing high risk of CS following AMI.Prolonged FMC time and the presence of left main and/or multivessel lesion are independent risk factors for the development of CS.The optimal revascularisation strategy can improve the clinical outcome of patients with CS.

Objective To investigate the pathogenesis of familial systemic lupus erythematosus (SLE), by analyzing the gene expression profile of peripheral blood in a family with 2 SLE patients and their first-degree relatives. Methods Total RNA was extracted from peripheral blood cells of normal subjects and SLE patients. Then, synthesis double strand cDNA template from total RNA, transcription of cRNA probe with Biotin labeling, hybridization of probe with Microarray, binding of Streptavidin to Biotin, amplification with First Antibody, further amplification with Cy3-Conjugated Second Antibody, detection of Cy3 dye with ScanArray 5000 were performed. With QuantArray microarray analysis software, the scan image information was converted into numeric data. With GeneSpring microarray analysis software, cluster analysis was done to find interested genes. Results Over 3000 target genes were analysed. Fifty-nine genes differentially expressed in familial SLE patients and controls were identified. Among them, 34 genes were up-regulated and 25 genes were down-regulated. These differentially expressed genes identified in two familial SLE patients were almost identical to those found in other sporadic SLE patients. Among 34 expression increasing genes, 22 were up-regulated in SLE sisters and unaffected sisters; among 25 expression decreased genes, 17 genes down-regulated in SLE sisters and unaffected sister. Cluster analysis showed that patients were clearly separated from controls and their unaffected sisters based on their gene expression profile. These results showed that in familial SLE, multiple genes were responsible for susceptibility to SLE, and clinically unaffected relatives shared some lupus susceptibility genes with their clinically affected relatives, in addition environmental factors were probably necessary to trigger disease. Conclusion These results indicate that high-density oligonucleotide microarray has the potential to explore the heredity in SLE families.

This study is to evaluate the effects of Shenmai injection on the temporal alterations of action potential (AP), early afterdepolarization (EAD) and delayed afterdepolarization (DAD) in papillary muscles. The action potentials were recorded by a glass electrode. APD at 90% repolarization (APD9 ) was measured, and spontaneous EAD and DAD were observed. The results show APD90 was significantly prolonged in model group compared with sham-operated group, whereas it was remained unchanged in Shenmai injec- tion treatment group and amiodarone group. The spontaneous EADs and DADs were frequently visible in model group. In conclusion, EAD, DAD and trigger activities increase gradually during pathological progression of rat cardiac hypertrophy, and Shenmai injection could improve the action potential change in rat cardiac hypertrophy.
Action Potentials ; drug effects ; Animals ; Blood Pressure ; drug effects ; Cardiomegaly ; physiopathology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Heart Ventricles ; drug effects ; physiopathology ; Injections ; Male ; Papillary Muscles ; drug effects ; physiopathology ; Rats ; Rats, Sprague-Dawley

The aim of this study is to prepare hyaluronic acid (HA) modified core-shell liponanoparticles (pHA-LCS-NPs) as gene delivery system and investigate its gene transfection efficiency in human retinal pigment epithelium (ARPE-19) cells in vitro. The pHA-LCS-NPs was prepared by firstly hydrating dry lipid film with CS-NPs suspension to get LCS-NPs, then modifying the lipid bilayer with HA by amidation reaction between HA and dioleoyl phosphatidylethanolamine (DOPE). Its morphology, particle size and zeta potential were investigated. XTT assay was used to evaluate the cell safety of different vectors in vitro. The gene transfection efficiency of pHA-LCS-NPs modified with different contents of HA was investigated in ARPE-19 cells with green fluorescent protein (pEGFP) as the reporter gene. The results showed that the obtained pHA-LCS-NPs exhibited a clear core-shell structure with the average particles size of (214.9 +/- 7.2) nm and zeta potential of (-35 +/- 3.7) mV. The 24 h cumulative release of gene from pHA-LCS-NPs was less than 30%. After 48 h incubation, gene transfection efficiency of pHA-LCS-NPs/pEGFP was 1.81 times and 3.75 times higher than that of CS-NPs/pEGFP and naked pEGFP, respectively. Also no obvious cytotoxicity was observed on pHA-LCS-NPs. It suggested that the pHA-LCS-NPs might be promising non-viral gene delivery systems with high efficiency and low cytotoxicity.
Cell Survival ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Hyaluronic Acid ; chemistry ; pharmacology ; Lipids ; Nanoparticles ; Particle Size ; Phosphatidylethanolamines ; chemistry ; pharmacology ; Retinal Pigment Epithelium ; drug effects ; Transfection