Objective Valsartan, the angiotensin Ⅱ type 1 receptor blocker, is recently proved to reduce urinary albumin at the microalbuminuria stage in human diabetic nephropathy without altering glucose metabolism. But the pathway is still uncertain. In present study, we examined the changes of adrenomedullin receptor (ADMR) mRNA and protein expressions in the renal cortex of diabetic rats to investigate the protective effects of valsartan on an experimental model of diabetic renal injury. Method The SD rats were randomly divided into following groups: normal rats, STZ-induced diabetic rats, and diabetic rats treated with valsartan. STZ-induced diabetic rats were treated with valsartan (10mg/kg body weight) or vehicle for 8 weeks. The expressions of ADMR mRNA in renal cortex were analyzed by RT-PCR, as well as ADMR protein expressions were detected through western blot. Results We found (1) Valsartan treatments reduced urinary albumin excretion in 24h, compared with the untreated. But no notable difference was seen in HbA1c and blood sugar of diabetic rats between the two groups. (2) Valsartan treatments increased the expressions of ADMR mRNA and protein in diabetic rats renal cortex. Conclusion These results indicate that valsartan treatment can upgrade the expressions of ADMR in the renal cortex of diabetic rats. It may be one of renal protective pathways of Ang Ⅱ type 1 receptor blocker.

Objective To explore the relationship between transforming growth factor-β1 (TGF-β1) and obesity-related glomerulopathy (ORG),and to analyze the possible mechanism for ORG and the new approach to its treatment.Methods Based on their body weight,30 SD rats were randomly divided into 2 groups : the normal control group (15 rats) fed with common food and the ORG model group (15 rats) fed with fat-enriched diets.The rats were sacrificed at the end of the 8th week,and their kidneys were taken out.Immunohistochemistry was used to detect TGF-β1 protein expression.Real time (RT)-polymerase chain reaction (PCR) was used to extract and detect the expression of TGF-β1 mRNA,and Western blot was applied to examine the expression of TGF-β1 protein.The findings were analyzed by using SPSS 13.0 software.Results Compared with the control group, qualitative TGF-β1 expression in ORG model group were significantly increased detected by immunohistochemistry mainly in renal tubules and interstitium.The average absorbance value of the control group and the model ORG group was 0.040-0.013,0.171 ± 0.084, respectively.The difference was statistically significant(P ＜ 0.05).The expression of TGF-β1 mRNA detected by RT-PCR was also increased compared with that of the control group(4.4 vs 0.6).The difference was statistically significant(P ＜ 0.05).The protein expression of TGF-β 1 examined by Western blot showed that it was more than that in the control group(4.3 vs 0.4).The difference between the control group and ORG model group was statistically significant(P =0.002).Conclusions The expression of TGF-β 1 in kidneys of ORG model rats increased, which not only indicates it can participate in ORG's occurrence and development, but also provide the basis to find out the mechanism and the approach to treatment.

According to bias in codon choice of Pichia pastoris, The phytase phyA gene from Aspergillus niger N25 was mutated without changing its amino acid sequence. The expression plasmid pPIC9k-phyAm was constructed and transformed into GS115 strain. Positive clones,of which the chromosomes were integrated with phyA gene,were identified by the phenotype and PCR. SDS-PAGE analysis suggust that the size of enzyme protein of the expression product was about 70.15kDa.Southern blotting analysis to the yeast transformants showed that phyA gene was intergrated into the chromosome genome. The phytase activity of PP-NP m-4-4 with codons optimized reached 136 000U/ml in malt wort culture medium after being induced with 36h, which was the 2.8 times of the original strain PP-NPm-8.

Objective: To optimize the expression of recombinant human monocyte chemoattractant protein (rhuMCP-1) in E.coli DE3. Methods:With NBS-MICROS 15 L T.DR fermentor, pGEX-IN/huMCP-1 was constructed by our laboratory. Four parameters including pH,temperature,agitation rate and concentration of IPTG were studied by orthogonal experimental design. Results: It was found that the expression level was greatly affected by the amount of dissolved oxygen. This indicated that the agitation rate and ventilation amount were the most important parameters during fermentation. Examined by SDS-PAGE and gel scanning, the expression level of total protein was over 40% when agitation rate was 300 r/min and ventilation amount was 10 L/min. Conclusion: A method for high-level expression of huMCP-1 on pilot-scale is established, and it will be useful for large-scale industrial production of target protein.

Hepatitis B surface antigen (HBsAg) carrying preS sequences could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. Here we report the success in achieving efficient and stable expression of hepatitis B virus S antigen and preS1 epitope fusion protein (S/preS1) in CHO cells. The HMRCHEF53u/Neo-S/preS1 expression vector carrying S/preS1 gene was constructed and transfected into CHO-S cells. A stable and high-expression CHO cell line, named 10G6, was selected by ELISA and limiting dilution analysis. Western blotting analysis showed S/preS1 expressed from 10G6 cells possessed both S and preS1 antigenicity. 10G6 cells displayed characters of favorable growth and stable S/preS1 expression in repeated batch cultures as evaluated by viable cell density, viability and S/preS1 concentration. And cultivation of 10G6 cells in fed-batch mode resulted in S/preS1 production at 17-20 mg/L with viable cell density at 7 x 10(6)-10 x 10(6) cells/mL.
Animals ; CHO Cells ; Cricetulus ; Epitopes ; biosynthesis ; genetics ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Vaccines ; biosynthesis ; genetics ; Hepatitis B virus ; Protein Precursors ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Bacterial Typing Techniques ; Genotype ; Minisatellite Repeats ; Plague ; Yersinia pestis ; genetics

The treatment results of radiofrequency ablation (m) were analyzed in l5 patients with the complicated Supraventricular tachycardia(SVT)． The causes of SVT were dual accessory pathway(AP) in 9 patients, single M with slow-fast atrioventricular junctional reentrant tachycardio (AVJRT) in 5 patients and single AP with slow fast/fast-slow AVJRT in l patient． Results: 24 APs and 6 slow pathways of atrioventricular node were abl a ted． All patients were successful． During the follow-up of 6～48 months, there were no recurrence in l5 patients, also there were no occurrence of atrioventricular block and other complications． Conclusion: Using the proper methods of m of the complicated SVT would have a h igher successful rate, a lower recurrent rate and lower complications．

OBJECTIVETo observe the effects of Danlou Tablet (DT) on inflammatory reaction, and expressions of lipoprotein-associated phospholipase A2 (LP-PLA2), secretory phospholipase A2 (sPLA2), and to analyze potential mechanisms.

METHODSForty male Wistar rats were randomly and equally divided into five groups, i.e., the normal control group, the model group, the Western medicine (WM) group, the low dose DT group, the high dose DT group, 8 in each group. Rats in the normal control group were fed with basic forage for 12 successive weeks, while AS rat model was established in rats of the other four groups by feeding high fat and sugar forage plus intraperitoneal injection of vitamin D₃. Normal saline, atorvastatin calcium suspension (at the daily dose of 1.8 mg/kg), low dose DT suspension (at the daily dose of 450 mg/kg), and high dose DT suspension (at the daily dose of 900 mg/kg) were administered to rats in the model group, the WM group, the low dose DT group, the high dose DT group respectively by gastragavage for 8 successive weeks. The general condition of all rats was observed. Rats were sacrificed after gastric administration and their serum collected. Serum levels of lipids (TC, TG, HDL-C, LDL-C) and inflammatory factors [IL-6, TNF-α, monocyte chemoattractant protein 1 (MCP-1), oxidized low-density lipoprotein (ox-LDL), lipoprotein-associated phospholipase A2 (LP-PLA2), secretory phospholipase A2 (sPLA2)] were detected. Pathological changes of thoracic aorta were observed by HE staining. Protein and gene expressions of LP-PLA2 and sPLA2 in thoracic aorta were measured by Western blot and real-time fluorescent quantitative PCR respectively.

RESULTSCompared with the normal control group, rats in the model group were in low spirits and responded poorly. Typical atherosclerotic plaque could be seen in thoracic aorta of rats in the model group. Serum levels of TC, TG, LDL-C, IL-6, TNF-α, MCP-1, ox-LDL, LP-PLA2, and sPLA2 significantly increased (P < 0.05); protein and gene expressions of LP-PLA2 and sPLA2 in rat thoracic aorta increased (P < 0.05) in the model group. After 8 weeks of intervention, rats in 3 medication groups appeared active, and HE staining showed subsidence of plaque in rat thoracic aorta. Compared with the model group, serum levels of TC, TG, LDL-C, IL-6, TNF-α, MCP-1, ox-LDL, and LP-PLA2 decreased in 3 medication groups (P < 0.01, P < 0.05); serum sPLA2 level decreased, protein and mRNA expressions of LP-PLA2 and sPLA2 in rat thoracic aorta decreased in the WM group (P < 0.01, P < 0.05); protein and mRNA expressions of LP-PLA2 in rat thoracic aorta significantly decreased in the low dose DT group (P < 0.01, P < 0.05), and those of LP-PLA2 and sPLA2 decreased in the high dose DT group (P < 0.01, P < 0.05).

CONCLUSIONDT could fight against inflammatory reaction and AS possibly through inhibiting LP-PLA2 expression and reducing ox-LDL production.

1-Alkyl-2-acetylglycerophosphocholine Esterase ; blood ; Animals ; Aorta, Thoracic ; pathology ; Atherosclerosis ; drug therapy ; Chemokine CCL2 ; blood ; Drugs, Chinese Herbal ; pharmacology ; Inflammation ; drug therapy ; Interleukin-6 ; blood ; Lipids ; blood ; Lipoproteins, LDL ; blood ; Male ; Phospholipases A2 ; blood ; Plaque, Atherosclerotic ; Random Allocation ; Rats ; Rats, Wistar ; Tablets ; Tumor Necrosis Factor-alpha ; blood

Objective To observe the expression of Neuropilin (NRP)-1, 2, to calculate micro vessel density (MVD) in colorectal cancer, paraneoplastic tissues, normal tissues, and to investigate the correlation between NRP-1, 2 and MVD to understand the role of NRP-1, 2 in the process of tumor angiogenesis. Methods Expression of NRP-1 and NRP-2 were studied by immunohistochemistry in 66 specimens from colorectal cancer, paraneoplastic tissues and normal tissues. MVD was assessed based on CD105 immunohistochical staining. Results (1) The positive expression of NRP-1, 2 in colorectal cancer, paraneoplastic tissues, normal tissues were 71.2%, 25.8%, 0;80.3%, 15.2%, 0 respectively. There was a statistical difference between them (P<0.05). (2)The quantity of MVD in colorectal cancer, paraneoplastic tissues and normal tissues were 35.682±5.542, 14.485±3.301 and 6.864±1.771 respectively. There was a statistical difference between them (P<0.05). (3) Expression of NRP-1, 2 had a good correlation with the tumor size, infiltrating depth, lymph node metastasis, Dukes stag-ing (P<0.05), but no correlation was found with the tumor position, histological types, differentiation degree and the gender or age of patients (P>0.05). (4) The MVD value had no correlation with the gender or age of patients, tumor size, position, histological types (P>0.05), but had a good correlation with infiltrating depth, lymphatic metastasis and Dukes staging (P<0.05). (5) There was a positive correlation between NRP-1, 2 and MVD value (P<0.01). Conclusion NRP-1, 2 may play an important role in the angiogenesis of the colorectal cancer, and it correlates with the invasion and metastasis of colorectal cancer closely.

Objective: To study the therapeutic effects of combined treatment with Chinese and western medicine in children with cerebral palsy (CP). Methods: 104 CP children were treated mainly with physiotherapy and acupuncture, together with occupational therapy, sensory integration and electrical stimulation. Gross Motor Function Measure (GMFM) was used to assess the effects of treatment. Results: After treatment for 3.29±0.80 months, the majority of CP children improved their IQ and limb symptoms, with decreased abnormal posture, increased muscle strength and improved motor developmental function. GMFM88 and GMFM66 scores had significantly increased after treatment. Conclusions: Combined treatment with Chinese and Western Medicine has confirmed effects on CP children.