The effects of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) on proliferation of bovine coronary artery endothelial cells (BCAEC) and smooth muscle cells (BCASMC) were studied in vitro. BCAEC and BCASMC were isolated and cultured and divided into control group, VEGF (50ng/ml) group and HGF (50ng/ml) group. Cells proliferation was measured using MTT method. The results showed that the OD values of control, VEGF, and HGF group in BCAEC cultures were 0.23?0.02, 0.58?0.10, and 0.42?0.12, respectively, and those in BCASMC were 0.31?0.08, 0.45?0.09, and 0.40?0.11, respectively. The proliferation ratios of BCAEC and BCASMC induced by HGF were 152.2%?33.8% and 45.2%?25.3%, respectively, and that by VEGF were 82.6%?18.7% and 29.0%?20.4%, respectively. The results suggested that HGF could promote proliferation and migration of BCAEC and BCASMC, while VEGF could promote proliferation of BCAEC but not BCASMC. The effect of HGF on BCAEC was stronger than that on BCASMC, and the induction strength of HGF was higher than VEGF.

[Objective] To explore the effect of the method of activating spleen to solidify superficial on intestinal microecology of children recurrent respiratory tract infection.[Method] Select 60 cases aged 1~12 of Qi deficiency type of lung and spleen in recurrent respiratory tract infection,randomly divide them into treatment group 30 cases and control 30;the treatment one takes TCM Fugangning formula orally for activating spleen to solidify superficial;the other takes orally BIFICO,for 12 w;compare their Bifidobacteria(B) and Escherichia coli(E) changes and B/E value after treatment,compare it to the control one's.[Result] After treatment,in the dejecta samples,the B content was higher than that in control one(P0.05);the B and E content and B/E were lower than normal group(P0.05),while control group was more than the normal group(P

Objective To evaluate the accuracy of 99mTcO4- gastrointestinal imaging in the diagnosis of Meckel’s diverticulum in children. Methods The clinical data of 99mTcO4-imaging, surgery and pathological results of 95 children with gastrointestinal bleeding were retrospectively analyzed. Results Forty-four cases of 95 patients had positive ifnding of 99mTcO4-ectopic gastric mucous membrane imaging, and positive rate was 46.3%. In the positve cases 52.3%cases (23/44) were diagnosed of intestinal heterotopic gastric mucosa, 47.7%cases (21/44) were suspected of intestinal heterotopic gastric mucosa. The sex ratio (males to females) was 7.8:1. Thirty-seven positive cases underwent operation;among them, 35 cases were found to have Meckel’s diverticulum and 1 case had duplication of the digestive tract;Diverticulum lesion was not found in 1 case. The samples of 35 cases diagnosed surgically of Meckel’s diverticulum were examined by pathologic histology. All 35 cases were conifrmed as Meckel’s diverticulum and ectopic gastric mucosa epithelium, including 2 cases with ulcer formation, 1 case with small patches of pancreatic tissue, 1 case with ileocecal junction of suppurative enteritis and 1 case with chronic appendicitis. Conclusions 99mTcO4-radionuclide imaging technique is of high value for the non-invasive diagnosis of children’s Meckel’s diverticulum.

ObjectiveTo evaluate the nephroprotective effects of transplanting metanephric mesenchymal cells (MMCs) into the renal subcaspsule of rats with acute tubular necrosis (ATN) induced by gentamicin. MethodsMMCs were expanded in culture and immunocytochemistry was used to characterize the cells. After gentamicin-induced ATN, fluorescence-labeled cells were transplanted and traced in kidney tissues by fluorescence microscopy. Serum creatinine (Scr) and N-acetyl-b-D-glucosaminidase (NAG) were tested. Kidney pathology was studied by hematoxylin-eosin staining. Apoptosis was examined by the TUNEL assay. Ki-67 and Bcl-2 expression was examined by immunohistochemistry. ResultsMMCs were expanded in culture and the phenotype of the cells was vimentin-positive and keratin-negative. Compared with other ATN groups, in the MMCs-treated group, Scr and NAG clearly decreased[14d Scr: (101.38±20.46) μmol/L vs (248.78±23.15), (252.98±33.52), (229.08±18.18) μmol/L;NAG: (14.83±7.74) U/L vs (33.33±14.88), (29.62±10.54), (30.22±10.94) U/L, P<0.05, respectively];the histopathoiogic lesion scores were lower (P<0.05);the Ki-67 antibody and apoptosis of renal tubular epithelial cells were improved or reduced respectively;the expression of Bcl-2 protein was up-regulated (P<0.05). ConclusionThe subcapsular transplantation of MMCs can ameliorate renal function and repair kidney injury.

Objective To evaluate the clinical effect of popliteal artery perforator-based sural neurovascular flap for repairing soft tissue defects in middle and distal upper leg.Methods Between December,2011 to September,2015,18 cases with skin soft tissue defects on the middle and distal upper leg were treated with popliteal artery perforator-based sural neurovascular flap,in which 12 cases were males,and 6 cases were females.The age was from 24 to 55 years,with the average age of 35.9 years.The size of tissue defects ranged from 3.0 cm × 3.0 cm to 7.0 cm × 9.0 cm.Results All flaps survived completely in 18 cases,1 case of marginal infection heated after dressing,and the rest cases in Ⅰ healing.The outline and function of survived flap were satisfactory during 3-16 months follow-up,with two-point discrimination of 4.0-10.0 mm.Conclusion The anterograde sural neurovascular flap based on the popliteal artery perforator provides a practical option for covering tissue defects in middle and distal upper leg.This flap is characteristiced by reliable blood supply without sacrificing main vesses,good contour and texture,in addition,the operation is easy of handling.

The Y118A、Y118F、Y118T、S378A、S378T、H310A、H310R mutants of Candida albicans sterol 14?-demethylase (CACYP51) were constructed and heterologously expressed in D12667, the reconstructed strain with the deletion of CYP51 gene of the Y12667. With the strains obtained and microsome enzymes separated, the western blot and the ultraviolet absorption spectrophotometry were used to qualitative and quantitative detect the expressed protein, the GC-MS was used to detect the metabolism activity of the protein. The results showed that, the target protein expressed successfully in the reconstructed strains, with the expression level up to 25% of the total microsome proteins. The results also showed that, the wild type protein had the catalytic activity to its nature substrate. While after alteration the wild gene with Y118A、Y118F、Y118T、S378A、S378T、H310A、H310R by a single base substitution, the catalytic activity of protein markedly decreased respectively. So the wild type and mutation CYP51 were expressed successfully in Saccharomyces cerevisiae and the expression products preserved the activity to metabolism their nature substrate.

Objective To investigate the therapeutic effect of acupoint application combined with Daqinglong Decoction (DD) for children asthma with the syndrome of exterior cold and interior heat. Methods One hundred and five children with mild and moderate persistent asthma were evenly randomized into combination group, DD group and Montelukast group. The three groups were separately given acupoint application combined with oral use of DD, oral use of DD, oral use of Montelukast, respectively. The treatment for the three groups lasted for 12 weeks. The scores of traditional Chinese medical syndromes in the three groups were recorded before and after treatment. The control of asthmatic symptoms, acute asthmatic attack frequency, and the course of disease were evaluated at the end of follow-up week 12. Results (1) At the end of treatment week 4 and 16, the syndrome scores were obviously decreased in the three groups (P<0.05 compared with those before treatment). At the end of treatment week 4, the decrease of syndrome scores in the combination group was superior to that in DD group (P<0.05), but did not differ from that in Montelukast group (P>0.05) . At the end of follow-up week 12, the decrease of syndrome scores in the combination group was superior to that in DD group and Montelukast group (P<0.05). (2) The effect of the combination group on controlling the asthmatic symptoms was superior to that of the other two groups ( P<0.01). ( 3) The combination group had lower acute asthmatic attack frequency and shorter disease course than DD group and Montelukast group ( P<0.01). Conclusion Acupoint application combined with oral use of Daqinglong Decoction is effective for children asthma with the syndrome of exterior cold and interior heat by relieving symptoms, enhancing the control of asthma, reducing acute asthmatic attack frequency and shortening disease course.

Objective:To understand the expression levels of Tim-3,a new proinflammatory factor in the early stages of Echinococcus granulosus infection in mice.Methods: BALB/c mice were infected with E.granulosus.Peritoneal macrophages and spleen cells were collected at 1,5,9 and 13 days post-infection.At different time points ,the levels of Tim-3 in peritoneal macrophages and spleen CD3+lymphocyte subsets were detected by FCM , and the relative expression of TLR 4 mRNA was detected by qRT-PCR.Results:There was no significant difference in the expression levels of Tim-3 of CD3+spleen lymphocyte subsets between E.granulosus group and control group (P>0.05).The expression levels of Tim-3 of spleen macrophages (9,13 days) and peritoneal macrophage (5,9,13 days) were much higher in E.granulosus infected group than those in control group with statistical significance (P<0.05).The numbers of macrophages were no change.Compared with control groups,the relative expression of TLR4 mRNA at 1 day post-infection was statistically higher in E.granulosus infected group ( P<0.05 ).Conclusion:During early stage of E.granulosus infection in mice,the levels of Tim-3 expression are upregulated,while the expression of TLR4 are downregulated,which may inhibit the function of macrophages resulting in host-immunity-defensive-system inhibition and immune tolerance of E.granulosus to host.

Objective:To investigate the effects of TGF-β1 on T lymphocytes of BALB/c mice infected with Echinococcus granulosus( E.granulosus ) in vitro.Methods: The inhibitor group:the spleen cells of BALB/c mouse were co-cultured with E.granulosus and SB525334.The control group:the spleen cells of BALB/c mouse were co-cultured with E.granulosus and PBS.The blank group:the spleen cells of BALB/c mouse were co-cultured with RPMI-1640 medium and SB525334.The lymphocytes were collected at 48 h post-infection.The T lymphocyte subsets, the number of CD4+CD25+T cells, the number of NK cells, and the expression of NKG2D receptor were detected by flow cytometry.The NK cell activity was determined with the lactate dehydrogenase leakage assay(LDH).Results:The inhibition the TGF-β1 receptors resulted in the increase of in the number of CD4+T cells,the decrease in the number of CD8+T cells,the increase of in the ratio CD4+/CD8+T cells,the decrease of in the number of CD4+CD25+T cells,the increase in the expression of the NKG2D receptors,the increase in the lysis rate of Yac-1 cells by NK cells,and a positive cor-relation between the expression of activity receptor NKG2D and killing activity of NK, which were mediated by E.granulosus.Conclusion: The inhibition of TGF-β1 receptors can enhance the immune response of T lymphocytes against E.granulosus infection in vitro.

Objective To investigate the prokaryotic expression and immunoreactivity of BspE,a type Ⅳ secretion protein of Brucella,and the effect of recombinant protein BspE on cytokines.Methods According to the BspE gene of Brucella M5-90 published in GenBank,the gene fragments were synthesized by a company and then ligated into PUC57 vector for sequencing.The sequenced gene was cloned into a prokaryotic expression vector pET-28α and transformed.Induced expression was performed in E.coli DE3 competent cells.The obtained target protein was purified by a Ni-NTA affinity column,and its reactogenicity was analyzed by Western blotting.Mouse RAW264.7 cells were treated with 25 g/L BspE recombinant protein for 12,24,48 h,and the control group was treated with the same amount of BSA instead of BspE,and enzyme-linked immunosorbent assay (ELISA) was used to detect interleukin (IL)-1β level.Results The recombinant expresed plasmid of pET-28α-BspE was successfully obtained.The results of Western blotting showed a single band with a relative molecular mass of about 30.1 × 103,and the recombinant protein BspE had good reactogenicity,and IL-1β levels (ng/L)were significantly elevated by the recombinant protein BspE (12 h:43.27 ± 2.13 vs 30.24 ± 1.66,24 h:57.78 ± 3.44 vs 41.22 ± 1.22,48 h:72.52 ± 3.04 vs 46.77 ± 2.75,t =8.38,7.86,10.89,P ＜ 0.05).Conclusions BspE recombinant protein has better immunoreactivity and can increase the expression level of IL-1β in mouse macrophages.This study provides a scientific basis for the role of effector proteins in the pathogenesis of Brucella.