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.

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

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 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.

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

10.3969/j.issn.2095-4344.2013.25.014

Objective To study the prevalence of the mtDNA A1555G gene mutation in Chinese population with nonsyndromic hearing impairment.Methods PCR-RFLP,directional sequencing of PCR products were applied in 325 patients with nonsyndromic hearing impairment and 50 normal controls.Results The mutation rate of the mtDNA A1555G was 14.5% (47/325),28 of 47 cases were homozygosis,19 of 47 cases were heterozygosis.The same mutation was not detected in the control subjects.Conclusion The mutation rate of the mtDNA A1555G is relatively high in the Chinese NSHI patients,the mutation type includes both heterozygosis and homozygosis.

Objective:By analyzing the norm data results of the Wechsler Adult Intelligence Scale-Fourth Edition (WAIS-Ⅳ) in China,to prove the validity of the procedure and methods during the norming development.Methods:The whole process of the revision of WAIS-Ⅳ,the development of computer-assisted system and norm sampling plan,were introduced in more detail,and the distribution of actual norm data of 1757 cases was analyzed.Results:For area distribution,compared with planned sampling number,the number from North and Northeast China was statistically significant different (x2 =78.02,P ＜0.01).For age stages distribution,most of cases conformed to the requirements of sampling,except that some cases including high-level educational cases aged 16-17 years and above 65 years,and low-level educational cases aged 30-34 years were less than the planned sampling number.For gender distribution,male subjects were more,but there was no statistically significant difference between male and female subjects (x2 =228,P =0.131).For educational degree distribution,the sampling conformed to the requirements of sampling plan (x2 =2.74,P =0.603).For occupation,resident years and registered permanent residence,and the sample was basically representative.Conclusion:The process of the revision of the Chinese version of WAIS-Ⅳ is appropriate,and actual norm sampling basically conforms to planned sample distribution,providing the sufficient representativeness and reliability for national norm data of WAIS-Ⅳ.