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 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 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 investigate the imaging features of adrenal rest tumor.Methods Twelve patients of adrenal rest tumor proved by surgery or clinical diagnosis were retrospectively analyzed.Among these 12 patients,12 were examined with ultrasound,11 with MR and 1 with CT. MR and CT were performed without and with intravenous injection of contrast material.The imaging features of adrenal rest tumor were retrospectively summarized and the relevant literatures reviewed. Results The adrenal rest tumors were found in testis in 10 of the 12 patients,and in ovaries and broad ligament in the remaining two.The imaging features of the testicular adrenal rest tumor were summarized as following:all patients had bilateral testicular masses without change of the testicular contour. On ultrasonography,the lesions were hypoechoic, with some hyperechoic areas and appeared highly vascularized on Colour Doppler ultrasonography.The masses showed iso-density on plain CT,and avid enhancement on post-contrast CT images.The masses ranging in size from0.7 cm×1.0 cm×2.2 cm to 2.3 cm ×2.7 cm ×2.9 cm with uniform signal intensity,lobulated margin on MRI.They exhibited iso- or slight hyperintensity on T1 WI and hypointensity on T2WI relative to normal testicular parenchyma.The tumors showed intense enhancement on post-contrast MR images. No abnormality was detected with Colour Doppler uhrasonography and MR in 2 patients of adrenal rest tumor in ovaries and broad ligament. Conclusion Combining imaging features with the typical clinical history,the diagnosis of adrenal rest tumor could be suggested pre-operatively.

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.

BACKGROUND:Cleft palate repair with levator veli palatini retropositioning can achieve good effects on pharyngeal function and voice quality by dissecting the muscle and rebuilding the muscle function only at the incision, without opening the mucoperiosteal flap, OBJECTIVE: To investigate the clinical effects of cleft palate repair with levator veli palatini retropositioning versus traditional surgery treatment on speech disorder correction. METHODS:Totaly 176 children with cleft palate were randomly assigned into treatment group and control group, 88 cases in each group. Patients were subjected to traditional surgery treatment in the control group and levator veli palatini retropositioning in the treatment group. RESULTS AND CONCLUSION:The postoperative speech inteligibility in the treatment group and the control group were 81.8% and 54.5%, respectively, and there was a significant difference between the two groups (P < 0.05). After cleft palate repair, the folowing indicators were al improved in the two groups, including soft palate functional length at static state and when pronouncing, and soft palate elevation angle (P < 0.05), and moreover, these indicators were better in the treatment group than the control group (P < 0.05). Scores on high nasal tone and rhinorrhea tone were decreased significantly in the two groups after repair, and the scores were significantly lower in the treatment group than the control group (P < 0.05). These findings indicate that cleft palate repair with levator veli palatini retropositioning can effectively rebuild soft palate function and perfect velopharyngeal competence in patients, which is of great significance for speech disorder correction and voice quality recovery.

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 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.

Multi-component traditional Chinese medicines are an innovative research mode for traditional Chinese medicines. Currently, there are many design methods for developing multi-component traditional Chinese medicines, but their common feature is the lack of effective connection of the traditional Chinese medicine theory. In this paper, the authors discussed the multi-component traditional Chinese medicine design methods based on medicinal property combination modes, provided the combination methods with the characteristics of traditional Chinese medicine for the prescription combinations, and proved its feasibly with hypertension cases.
Animals ; Antihypertensive Agents ; administration & dosage ; chemistry ; Blood Pressure ; drug effects ; Drug Combinations ; Drug Therapy ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Humans ; Hypertension ; drug therapy ; physiopathology ; Medicine, Chinese Traditional ; Rats