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 .

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 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.

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 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.

Animals ; Anti-HIV Agents ; pharmacology ; HIV ; drug effects ; genetics ; physiology ; HIV Infections ; drug therapy ; immunology ; virology ; Humans ; Mucous Membrane ; immunology ; virology ; Virus Replication ; drug effects

Objective To investigate the effect of triiodothyronine on the expression of alpha subunit of C protein Co (Coa) gene in cultured rat cerebral cortex neurons. Methods Primary cultured neurons of rat were prepared from the cerebral cortex at embryonic day 19. The neurons were cultured in DMEM supplemented with 15% fetal calf serum. After 7 days, the neurons were cultured in different media; DMEM supplemented with 15% fetal calf serum (group A), DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group B), 0.5nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group C) , 5nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group D) and 50nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group E). The neurons were cultured for another 7 days and total RNA was extracted. Goa mRNA leves were measured by competitive RT-PCR. Results As compared with group A, Coa mRNA leves in group B, D and E were low (P 0.05). Conclusion T, has dual effects on the expression of Coa gene in cultured rat cerebral cortex neurons. T3 down-regulated Coa mRNA in cultured neuron. However, in the absence of T3, Goa mRNA was also very low. Thyroid hormone may exert their action on brain development by regulating the the expression of Goa gene.

Objective:To investigate the antimicrobial resistance spectrum of Escherichia coli isolated from different clinical sam-ples to guide the nosocomial infection control and rational use of antibiotics. Methods:Escherichia coli from clinical samples were isola-ted and tested by microbiological assay system of BD Phoenix 100 and susceptibility test card in 2013. The drug resistance was analyzed and compared retrospectively. Results: There were 936 strains of Escherichia coli isolated from our hospital in 2013. Escherichia coli was resistant to common antibiotics at high level. Escherichia coli strains isolated from different samples varied in the drug susceptibili-ty. The resistance rate of the strains from sputum samples was higher than that from the other samples. Conclusion:The status of drug resistance of clinical isolated Escherichia coli to antibiotics is very serious. Escherichia coli isolated from different samples show different drug resistant features, and the antimicrobial therapy should be considered according to the antimicrobial susceptibility tests.

Objective To investigate the results of graft repair of peripheral nerve defects using small intestinal submucosa(SIS) with crude Schwann cells. Methods Thirty-six male SD rats were randomly divided into three groups,with 12 in each group.Firstly,12-mm gaps of sciatic nerve were made in rats and then treated respectively with porcine SIS(group A),SIS compounded with crude Schwann cells(group B) and auto-nerve grafting(group C) in the three groups.The grafts were harvested 16 weeks postoperatively to evaluate the results of nerve regeneration through histological examination, triceps surae weight,computerized imaging analysis,Trueblue retrograde tracing and transmission electron microscopy. Results Nerve regeneration was confirmed to have extended through the gaps according to the results of histological examination,Trueblue retrograde tracing and transmission electron microscopy.Furthermore,in the respect of triceps surae weight,quantity of axis cylinder per area and percentage of neural tissue,the results of group B and group C were superior to those of group A with a significantly statistical difference(P0.05). Conclusion SIS compounded crude Schwann cells was able to achieve the outcome of auto-nerve grafting and was a promising replacement graft.