Objective To obtain abundant human betacellulin(BTC) with biological activity. Methods The whole mature protein coding sequence of BTC gene was amplified by polymerase chain reaction(PCR) method applied to human pancreatic ?-cell tumors cDNA.The fragment was cloned into prokaryotic expression vector pET32a(+) plasmid.The recombinant plasmid was transformed into E.coli BL21 and the fusion protein was expressed under isopropyl-beta-D-thiogalactopyranoside(IPTG).The fusion protein was purified by Ni2+ affinity chromatography.SDS-PAGE and Western blot were employed to determine the expression and purification of the expected protein.BTC was added to culture NIH3T3 cells for 5 days,and cell proliferation was detected by MTT. Results Lots of fusion protein were produced,and the purified protein can stimulate the proliferation of NIH3T3 cells. Conclusion The human BTC can be successfully obtained from the pET32a(+) system with the biological activity of stimulating the proliferation of NIH3T3 cells.

Objective To construct eukaryotic expression vector of human pancreatic duodenal homeobox 1(PDX-1) gene,and to detect its expression in NIH3T3 cell lines. Methods The whole coding sequence of PDX-1 gene was amplified by polymerase chain reaction(PCR) from human pancreatic-cell tumors cDNA.The fragment was inserted into eukaryotic expression vector pcDNA3.1 plasmid.The recombinant plasmid was verified by double digestion and DNA sequencing.The expression of PDX-1 gene in NIH3T3 cells was assayed by Western blot. Results The length of specific fragment amplified by PCR was 852 bp,and the recombinant plasmid pcDNA3.1-PDX-1 showed two bands of 5.5 kb and 852 bp by digestion using respective restriction enzymes BamHⅠand EcoRⅠ.The sequence of PDX-1 gene was approved or confirmed by blasting to GenBank.It was suggested that PDX-1 gene had been cloned into pcDNA3.1 vector correctly.Western blot showed that PDX-1 gene was expressed,which was detected 24 h after pcDNA3.1-PDX-1 plasmid was transfected into NIH3T3 cells. Conclusion The recombinant eukaryotic expression vector pcDNA3.1-PDX-1 was successfully constructed and expressed in NIH3T3 cell lines.

Objective To evaluate the efficacy of ultrasound?guided transversus abdominis plane ( TAP) block with different concentrations of ropivacaine for analgesia after cesarean section. Methods A total of 120 parturients, aged 24-31 yr, of American Society of Anesthesiologists physical statusⅠ or Ⅱ, weighing 64-73 kg, at 35 to 41 week gestation, scheduled for elective cesarean section, were randomly di?vided into 3 groups ( n= 40 each) using a random number table: 0?25% ropivacaine group ( group Ⅰ) , 0?20% ropivacaine group ( groupⅡ) , and 0?15% ropivacaine group ( groupⅢ) . A patient?controlled an?algesia pump was connected at the end of surgery, and the corresponding concentration of ropivacaine 1?5 mg∕kg was injected into the bilateral TAP under the guidance of ultrasound in each group. Visual analogue scale score was maintained ≤3. The number of attempt and the number of patients requiring rescue analge?sic were recorded at 48 h after surgery. The occurrence of TAP block?related complications was observed and recorded. Results Compared with groupⅢ, the number of attempt and the number of patients requi?ring rescue analgesic were significantly decreased in Ⅰ and Ⅱ groups ( P<0?05 ) . Compared with groupⅡ, the number of attempt and the number of patients requiring rescue analgesic were significantly de?creased in group Ⅰ (P<0?05). No TAP block?related complications were detected in the three groups. Conclusion Ultrasound?guided TAP block with 0?25% ropivacaine is helpful in improving the analgesic efficacy after cesarean section without serious complications.

Objective To investigate the prevalence rate of new-onset malignant tumors in type 2 diabetes mellitus patients .Methods Type 2 diabetes mellitus patients ( 1771 cases ) over 3 years since May 2006 in Xinhua Hospital were retrospectively recruited .The prevalence and distribution of different tumors were analyzed .Results ⑴Total prevalence of malignant tumors was 2.88%, and more common in male ( P <0.05 ) .The prevalence of different malignant tumors in human systems from high to low were digestive, urinary, and respiratory .⑵Age-dependent analysis from <50, 50~, 60~, 70~,and≥80 showed that the prevalence were 0.73%,1.93%,3.27%,3.58%, and 5.02% respectively .A significant difference was found in them ( P <0.05 ) .Conclusion The prevalence of new-onset malignancies is higher in type 2 diabetes mellitus , which is associated with age and sex .Tumor screening should be strongly recommended in such a group of the patients for their earlier diagnoses and treatments due to the un -typical clinical manifestations .

The abdominal visceral fat content in obese SD rats induced by high fat diet for 10 weeks was significantly higher than that in control group [(26±6 vs 13±3)g,P<0.01] ,along with increased CRP mRNA expression in abdominal visceral fat (0.901±0.085 vs O. 402±0.036, P<0.01). As compared with normal control group, in the high fat group the concentrations of CRP in portal vein [(743.8±95.8 vs 558.3 ±118.3) mg/L, P<0.01] and peripheral vein[(596.3±38.9 vs 485.8±30.2) mg/L,P<0. 05] were higher. The concentration of CRP in portal vein was significantly higher than that in peripheral vein in high fat diet group(P<0.01) ,but this was not evident in control group. These results suggest that the increased CRP expression in visceral adipose tissue may partially account for the elevation of serum CRP in obesity.

Objective To explore the impact of advanced glycation end-products ( AGEs) on matrix metalloproteinase 2 ( MMP-2) expression in cultivated INS-1 cells. Method INS-1 cells were cultivated and MMP-2 expression was analyzed. Glycated serum was prepared for incubating with INS-1 cell. Reactive oxygen species (ROS) was detected by flow cytometry. The intracelluar MMP-2 expression was analyzed by RT-PCR, realtime PCR and Western blot. The MMP-2 cDNA was expressed in cultivated INS-1 cells. Result The level of ROS treated with AGEs was significantly higher than that in the control( P<0.05 ) , and the levels of MMP-2 and its protein expressions turned out as well( P<0. 05). Conclusion The results suggest that MMP-2 was expressed in INS-1 cells. Increased MMP-2 expression in ?cells may be induced by AGEs, suggesting that MMP-2 might play an important role in oxidative stress-mediated islet injury.

Objective To evaluate the distribution of non-motor symptoms(NMS) and dominant factors impacting on the prevalence of NMS in Parkinson's disease(PD). Methods The NMS questionnaire(NMSQuest) was administered to 128 patients(PD group) and 128 healthy adults(control group).The distribution of items of NMS was analyzed between the two groups,and clinical characteristics were also collected to assess the dominant factors impacting on the prevalence of NMS. Results There were significant differences in the prevalence of NMS between the two groups(P

Objective To evaluate the clinical value of CHO-hTSHR cells in detecting thyroid stimulating antibody (TSAb). Methods The cAMP production and TSAb activity were measured and cal-culated by stimulating CHO-hTSHR cell line with IgGs of normal control group and Graves" disease ( GD )group. TSAb positive standard was set to more than the mean + 2SD of TSAb activities in control subjects.The positive percentage of TSAb activity in GD group was calculated. Results The cAMP production and TSAb activities of GD group were higher than those of normal group[( 353. 65±126. 34 ) pmol/L vs (237.21±77. 15)pmol/L, ( 149. 08±53. 26)% vs ( 100±32. 52)%, P <0. 05] . The value that higher than 165% was set to be positive for TSAb. The positive percentage of TSAb in GD group was 50% ( 14/28). Conclusion CHO-hTSHR cell line constructed by our group is suitable for detecting TSAb activity in the sera of patients with GD.

Objective To investigate the relationship between the serum apolipoprotein B (ApoB)/ ApoA-Ⅰ ratio and intracranial artery stenosis in young patients with ischemic stroke.Methods Patients with ischemic stroke aged from 18 to 45 were enrolled in the study.Brain CT angiography was used to evaluate the degree of intracranial artery stenosis,and the concentrations of serum total cholesterol,triglyceride,highdensity lipoprotein cholesterol,low-density lipoprotein cholesterol,ApoA-Ⅰ,and ApoB were detected.The ratio of ApoB/ApoA-Ⅰ was calculated.The Demographic and clinical characteristics of the intracranial artery stenosis group and the non-intracranial artery stenosis group were compared.Multivariate logistic regression analysis was used to identify the independent risk factors for intracranial artery stenosis in young patients with ischemic stroke.Results A total of 161 young patients with ischemic stroke were enrolled,including 89 in the intracranial artery stenosis group and 72 in the non-intracranial artery stenosis group.The constituent ratios of diabetes mellitus (20.2％ vs.6.9％;x2 =4.641,P =0.032),smoking (47.5％ vs.15.2％;x2 =15.121,P=0.001),hyperlipidermia (56.1％ vs.48.6％;x2 =4.197,P=0.040),as well as the radios in serum high-density lipoprotein cholesterol (1.29 ± 0.30 mmol/L vs.1.65 ± 0.34 mmol/L;t =7.131,P=0.002),ApoA-Ⅰ (1.49 ± 0.65 g/L vs.1.63 ± 0.23 g/L;t =2.751,P =0.001),ApoB (1.49 ± 0.65 g/L vs.1.63±0.23 g/L;t=2.751,P=0.001),and ApoB/ApoA-Ⅰ ratio (1.49±0.65 vs.1.63± 0.23;t =2.751,P=0.001) had significant differences between the two groups.Multivariate logistic regression analysis showed that diabetes (odds ratio [OR] 3.052,95％ confidence interval [CI] 1.186-7.856;P =0.021),smoking (OR 2.997,95％ Cl 1.456-6.172;P =0.003),hyperlipidemia (OR 4.745,95％ CI 2.108-10.668;P =0.001),ApoB (OR 4.861,95％ CI 3.029-7.802;P=0.001),and ApoB/ ApoA-Ⅰ ratio (OR 5.684,95％ CI 2.215-14.584;P=0.002) were the independent risk factors for intracranial artery stenosis in young patients with ischemic stroke,while HDL-C (OR 0.561,95％ CI 0.354-0.888;P=0.014) and ApoA-Ⅰ (OR 0.065,95％ CI 0.010-0.409;P=0.004) were the independent protective factors.After adjustment for hypertension,diabetes,smoking,hyperlipidemia,HDL-C,ApoA-Ⅰ,and ApoB,ApoB/ApoA-Ⅰ ratio was still an independent risk factor for intracranial artery stenosis in young patients with ischemic stroke (each increase of 1 standard deviation,OR 4.255,95％ CI 2.348-7.711;P=0.001).Conclusion ApoB/ApoA-Ⅰ ratio is an independent risk factor for intracranial artery stenosis in young patients with ischemic stroke.

Aim To study the effect of emodin on IL-8 secretion and NF-?B activation of HT-29 cells,and explore the molecular mechanism of emodin.Methods The cytotoxicity of emodin was assessed by WST;NF-?B activation was detected with co-focal microscopy by immunofluorescence;the production of IL-8 was investigated by ELISA.Results Emodin with the concentration of 10~80 ?mol?L-1 could decrease the mass production of IL-8 Secretion of HT-29 cells stimulated by IFN-?+LPS in a dose-dependent manner.Emodin with various concentrations could inhibit NF-?B activation dose-dependently.Conclusions Emodin inhibited IL-8 secretion and NF-?B activation of HT-29 cells stimulated by IFN-?+LPS.