Objective To construct human phage antibody library against hepatoma carcinoma.Methods Peripheral blood mononuclear cells(PBMCs) of patients with liver cancer were sensitized in vitro and transformed by Epstein-Barr virus(EBV).The genes of light chain and Fd of antibodies were amplified by RT-PCR.Fab genes were cloned into vector pComb3 and transformed into E.coli XLI-Blue by electroporation to construct the Fab-displaying phage antibody library.Results ELISA detection showed that 4 liver cancer patients' B cells transformed by EBV could produce specific antibodies to hepatoma carcinoma cell.Totally 13 types of light chain genes and 28 types of Fd genes were obtained by RT-PCR.The capacity of the primary phage library was 1.7?107pfu/mL.The percentage of recombinant clones was about 100%.Conclusion A human phage antibody library has been constructed successfully by means of EBV transformation technique.

Objective To investigate the clinical value of contrast-enhanced ultrasound(CEUS) in the diagnosis of postnatal placenta increta. Methods Twenty-six patients with postnatal placenta increta were examined by gray-scale and color Doppler ultrasound and CEUS. Then microvascular perfusion and enhanced features of lesions, myometrium and serous layer were observed. Arriving time (AT), time to peak intensity (TTP) and the lasting time of enhancement (LTE) were recorded. AT,TTP and LTE of enhanced lesions were compared with those of normal myometrium. Results Serous layer, lesions and adjcent myometrium,normal myometrium enhanced in turn. There was no obvious boundary between the lesions enhanced and adjcent myometrium. AT and TTP of the lesions enhanced were both less than those of normal myometrium ( P ＜0. 05). LTE of the part of lesions enhanced was more than that of normal myometrium ( P ＜0.05).Part of lesions never enhanced during the whole process. The serous layer of uterine was smooth and uninterrupted in 24 patients. These 24 patients all recovered after conservative treatment. The local serous layer adjcent lesions was not smooth, but no contrast agent leakage occurred in another 2 patients, and uterine lobectomy were performed in emergency because of massive hemorrhage during conservative treatment. Conclusions Microvascular perfusion and enhanced features of lesions,myometrium and serous layer could be showed clearly through CEUS.

Objective The present study investigated the expression of heat shock cognate protein 70 (Hsc70) in the synovial tissues and blood samples of patients with rheumatoid arthritis (RA) to determine the pathological role of this protein in the pathogenesis of the disease.Methods The expression of Hsc70 in synovial membranes was quantitatively analyzed by immunohistochemistry,real-time quantitative PCR and western blotting.The samples from osteoarthritis (OA) and ankylosing spondylitis (AS) were used as controls.The levels of Hsc70 in blood of patients with RA were determined using enzyme linked immunosorbent assay (ELISA) with the samples of the healthy subjects as controls.Statistical analysis was conducted with one-way ANOVA,LSD test and Spearmen's correlation.Results Immunohistochemistry showed that Hsc70 had significantly increased expression in synovial tissues of RA than in the samples of OA and AS.Real-time PCR and western blotting confirmed the above findings.ELISA detected significantly elevated level of Hsc70 in blood of patients with RA as compared with samples from the controls (P＜0.01).Conclusion The study suggests that the up-regulation of Hsc70 may be involved in the pathogenic process of RA.

Objectives To evaluate the effects of combined atorvastatin and probucol use on endothelial function in patients with acute coronary syndrome (ACS). Methods Thirty patients with ACS were randomized to receive atorvastatin (20 mg/d) and probucol (500 mg/d, combination group, n = 15) or atorvastatin (20 mg/d) alone (atorvastatin group) within 24 h after admission for 4 weeks. Endothelium-dependent flow-mediated dilatation (FMD) and endothelium-independent sublingual nitroglycerin-mediated dilatation (NMD) as well as the levels of lipids and C-reactive protein were assessed at baseline, 1 week and 4 weeks after therapy. Results Compared to baseline, the levels of total cholesterol, LDL-C and C-reactive protein were significantly reduced after 1 week and 4 weeks in both groups, FMD equally increased after 1 week in both groups (atorvastatin group: 3.75%±0.78% vs. 1.09%±0.44%, combination group: 3.67%±0.36% vs. 1.24%±0.37%, P<0.01). Post4 weeks therapy, FMD increase was significantly higher in combination group (3.67%±0.36% at 1 week vs. 6.85%±0.64% at 4 weeks, P <0.01) than that in atorvastatin group (3.75%±0.78% vs. 3.80%±0.31%, P=0.954). NMD also equally and increased over 4 weeks in two groups (P<0.01 vs. baseline). There was no correlation between the change in FMD/NMD and the changes in lipids or C-reactive protein levels. Conclusions The combined atorvastatin and probucol therapy early after ACS is superior to atorvastatin alone on improving endothelial function.

Objective To observe the effects of soluble epoxide hydrolase inhibitor t-AUCB on foam cell formation and cholesterol efflux in macrophage.Methods Mouse macrophages RAW264.7 were cultured and stimulated with ox-LDL (80 μmol/L) in the absence (group A) or presence of t-AUCB (1,10,50,100 μmol/L,group B) or t-AUCB (100 μmol/L) pretreated with PPARγ antagonist GW9662 (5 μmol/L,group C).The foam cell was identified by oil red O staining.The cholesterol efflux rates of 3 Hcholesterol in cells were measured by liquid scintillation counter.mRNA and protein expressions of ABCA1 were detected by real-time PCR or Western blot,respectively.Results Oil red O staining showed that t-AUCB (100 μmol/L) significantly inhibited foam cell formation which could be significantly reversed by GW9662 (all P ＜ 0.05).t-AUCB dese-dependently increased cholesterol efflux rates in mouse macrophage [(5.91+0.18)％ in group A,(7.03 ±0.33)％,(8.05 ±0.32)％,( 9.04 ±0.14)％,(10.06±0.85)％ in 1,10,50,100 μmol/L t-AUCB groups,all P＜0.05 vs.group A],which could be reversed by pretreatment with GW9662 [ (6.33 ±0.15)％ in 100 μmol/L t-AUCB + GW9662 group].t-AUCB also upregulated ABCA1 mRNA and protein expressions in a dose-dependent manner which could be significantly attenuated by pretreatment with GW9662.Condusion t-AUCB could inhibit foam cell formation by improving cholesterol efflux through activating PPARγ-ABCA1 pathway in macrophage.

Objective To further evaluate the safety and observe for unknown adverse reactions in the prelicensure trials of Anflu(split influenza virus vaccine)in children aged 6 months to 3 years old and 3 to 11 years old respectively.Methods This open clinical trial enrolled 100 healthy children in each of the two groups:6 months to 3 years old group and 3 to 11 years old group.The 6 months to 3 years group were vaccinated with two pediatric doses(0.25 ml/dose),28 days apart.The 3 to 11 years group were vaccinated with one adult dose(0.5 ml/dose).All the subjects were observed for 30 min after vaccination and had 3 follow-up visits at 24,48,72 h after vaccination. All subjects with adverse reactions were followed up till the symptoms resolved.Results The total adverse reaction rate was 6.0%(12/200).The occurrence rates of local reaction and systemic reaction were 1.0%(2/200)and 5.5%(10/200)respectively.For the younger group and older group,the adverse reaction rates were 8.0%and 4.0%respectively.Conclusion This vaccineis safe in children aged 6 months to 11 years old.

Objective To further evaluate the safety and observe for unknown adverse reactions in the prelicensure trials of Anflu(split influenza virus vaccine)in children aged 6 months to 3 years old and 3 to 11 years old respectively.Methods This open clinical trial enrolled 100 healthy children in each of the two groups:6 months to 3 years old group and 3 to 11 years old group.The 6 months to 3 years group were vaccinated with two pediatric doses(0.25 ml/dose),28 days apart.The 3 to 11 years group were vaccinated with one adult dose(0.5 ml/dose).All the subjects were observed for 30 min after vaccination and had 3 follow-up visits at 24,48,72 h after vaccination. All subjects with adverse reactions were followed up till the symptoms resolved.Results The total adverse reaction rate was 6.0%(12/200).The occurrence rates of local reaction and systemic reaction were 1.0%(2/200)and 5.5%(10/200)respectively.For the younger group and older group,the adverse reaction rates were 8.0%and 4.0%respectively.Conclusion This vaccineis safe in children aged 6 months to 11 years old.

OBJECTIVE: To identify possible risk factors for cerebral palsy (CP) in children. METHODS: A Population-based survey was conducted (including 92 CP cases) in 66 townships of 15 cities of Zhejiang Province from October to November, 1998. 184 of matched controls were selected for comparison. RESULTS: Factors identified which were statistically significant for risk of subsequent childhood Cerebral Palsy included some neonatal diseases, some maternal diseases, low birth weight (<2500 g), maternal irregular menstruation, toxic, substances during pregnancy, malnutrition during pregnancy,and paternal age. CONCLUSION: Several risk factors for Cerebral Palsy were identified. Their prevention may result in redduction of the incidence of Cerebral Palsy.

Objective To observe the effects of acupuncture therapy with finger on back-shu point on acid reflux and lower esophageal sphincter pressure (LESP) of the patients with gastroesophageal reflux disease (GERD). Methods Totally 120 patients of GERD were randomly divided into treatment group and control group through random number table method, 60 cases in each group. Patients in the treatment group were treated with the acupuncture therapy with finger on back-shu point, and patients in control group were treated with lansoprazole tablets and dispersible mosapride citrate for two weeks. Total percentage of acid reflux time, the long time acid reflux episodes, and the longest acid reflux time of two groups were observed six months after the treatment. At the same time, the LESP variation of two groups was followed up six months after the treatment. Results The total percentage of acid reflux time, the long time acid reflux episodes, and the longest acid reflux time decreased significantly in all patients after treatment (P<0.01), while the comparison between groups showed no significant difference (P>0.05). After treatment, LESP of two groups was significantly improved (P<0.05) than before treatment. After stopping treatment half a year, the treatment group had obvious difference (P<0.05) compared with before treatment, while the control group had no significant difference (P>0.05). Conclusion The acupuncture therapy with finger on back-shu point can reduce acid reflux, and achieve the goal of treatment of GERD by improving the lower esophageal sphincter pressure. The duration of improving LESP is longer.

Objective To observe the role of abatacept in the treatment of diabetic nephropathy in rats. Methods Type 2 diabetic nephropathy rat model was established, and was randomly divided into abatacept group and non-intervention group, 15 each. 15 normal rats were served as control group ( NC) . The abatacept group was given abatacept for 8 weeks. The control group was set at the same time period. Then the blood biochemical indexes, blood flow parameters of renal main artery, elasticity of renal parenchymal, HE staining of renal parenchymal and ultrastructure of podocytes were all evaluated. The expression of CD31, CD34, podocin, nephrin, and B7-1 in renal parenchyma were detected. Results Compared to the NC group, the fasting blood glucose, creatinine clearance rate, urine albumin excretion rate, and kidney hypertrophy index in the non-intervention and abatacept groups were significantly increased (P<0. 05);creatinine clearance rate, urine albumin excretion rate, and kidney hypertrophy index in the abatacept group were significantly lower compared to the non-intervention group (P<0. 05). There were significant differences in peak systolic velocity, end diastolic velocity, and mean velocity among these 3 groups, and these differences were ranked as non-intervention groupabatacept>control groups (P<0. 05). There were significant differences in the stiffness of kidneys among these three groups, with the differences ranked as non-intervention>abatacept>control groups (P<0. 05). HE staining and transmission electron microscope observation showed that there were obvious changes in the renal parenchyma of rats in the non-intervention group and the abatacept group, and the changes of kidney in the non-intervention group were worse than those in the abatacept group. CD31 and CD34 expression in kidney parenchyma of rats in the abatacept group and non-intervention group were higher than those in the control group, the abatacept group and non-intervention group>the control group (P<0. 05), while no significant differences between the abatacept group and non-intervention group in CD31 and CD34 expression (P>0. 05). There were significant differences in podocin and nephrin expression in kidney parenchyma of rats among these three groups, and these differences were ranked as the control>abatacept>non-intervention groups (P<0. 05). There were significant differences in B7-1 expression in kidney parenchyma of rats among these three groups, and these differences were ranked as the control