Six bacterial strains with malachite green decolorization ability were isolated from a sediment of aquaculture pond, and strain M6 was selected by further enrichment culture in nutrition broth with malachite green and decolorization rate comparison. The decolorization rate of strain M6 to malachite green was 97.14% in the conditon of 30?C and 150 r/min, and its morphology was observed by gram stain and electronmicroscopy, its physiological and biochemical characteristic was studied by ATB bacteria identification in-strument for identification of bacteria, and its 16S rDNA sequence was determined following PCR amplifi-cation, the sequence was aligned and the phylogenic tree was instructed with those bacterial strains of high identity with strain M6. In addition, its growth characteristics was also studied. The experimental results showed that strain M6 was gram negative and bacilliform with a flagellum at one end. Its size was 0.45 ?m ?0.84 ?m. Its colony produced on common agar plate appeared as round, light blue, dense, hard to choose; 16S rDNA sequence of strain M6 had high identity of 98%～99% with Pseudomonas sp. located in GenBank and strain M6 had the most close relative relation to Pseudomonas putida OW-16 (Locus number: DQ112328.1). Combined the results of the traditional morphological, physiological, biochemical character-istics and 16S rDNA sequence analysis, strain M6 was identified as Pseudomonas putida (Locus number: EU348741.1). Additionally, its growth curve in the condition of 30?C and 150 r/min was as follows: lag phase was 0～4 h, log phase was 4 h～64 h, stationary phase was 64 h～80 h, decline phase was after 80 h. Its best growth conditions were pH 7 and 30?C,and in the rotational speed of 50 r/min to 250 r/min. Its concen-tration increased with the increase in rotational speed.

12,9.41%(148/1578);RDQ≤12,91.58%(1430/1578).Logistic multiple regression analysis of gastroesophageal reflux correlation factor studied:OR= 2.781.Conclusions The results showed:there were close correlation of high salt diet and GERED.

Objective To explore the influence of the lentiviral vectors-mediated mouse genetic engineering regulatory T cells (Treg) infused after allogeneic bone marrow transplantation (alloBMT) on graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect in mice.Methods Lentivirus-mediated expression of Forkhead box P3 (Foxp3) transformed CD4 + CD25- T cells from Balb/c mice into engineered Tregs in vitro. An allo-BMT model of Balb/c→C57BL/6 mice was established. The recipients were given lethal X-ray total body irradiation before transplantation.Mice were randomly assigned into five groups and each group contained 10 recipients: (1) The recipients in radiation group were injected with 0.2 ml RPMI 1640; (2) The recipients in leukemia control group were injected with 5 × 106 donor bone marrow cells and 500 mouse T-cell leukemia/lymphoma cells (EL4 cells); (3) The recipients in transplantation control group were injected with 5 × 106 donor bone marrow cells and 5 × 106 splenocytes plus 500 EL4 cells; (4) The recipients in engineering Treg group were injected with 5 × 106 donor bone marrow cells, 5 × 106 splenocytes and 500 EL4 cells plus 5 × 106 genetic engineering Treg; (5) The recipients in empty vector control group were injected with 5 × 106 donor bone marrow cells, 5 × 106 splenocytes and 500 EL4 cells plus 5 × 106 empty vector-transduced CD4+ CD25- T cells. Survival time, clinical GVHD score or histopathological analysis (skin, liver and small intestine) were observed after allo-BMT. Chimerism of bone marrow cells from recipients survived for 60 days after transplantation was measured. Results The mean survival time in radiation group, leukemia control group, transplantation control group,engineering Treg group and empty vector control group was ( 10. 3 ± 1.5), (20. 7 ± 1.9), (26. 0 ±4.3), (49. 0 ± 17. 7) and (24. 4 ± 4. 1 ) days respectively. The survival time in engineering Treg group was significantly prolonged as compared with other groups as judged by the log-rank test (P＜0. 05).Histopathological analysis in several target organs (skin, liver and small intestine) confirmed the presence of severe GVHD in transplantation control group and empty vector control group. No histological signs of GVHD or leukemia were observed in recipients in engineering Treg group and clinical GVHD scores in this group were significantly decreased as compared with transplantation control group and empty vector control group. Conclusion Co-injection of genetic engineering Treg can efficiently prevent recipients from lethal GVHD without affecting GVL activity during allo-BMT in mice.

To observe the effects of Danshen-containing serum on SuFu and DYRK2 expression in the HSCs stimulated by leptin. SD rats (n = 60) were used to make danshen-containing serum by gastric perfusion for ten days with Danshen water decoction, normal saline and colchicine. The HSCs that were cultured in vitro would be stimulated for 24 hours by leptin (100 μg x L(-1)) except blank control group, after being intervened, the drug serum in each group would be cultured at 37 degrees C in 5% incubator. The cells would be collected after 24 hours, then the effects of danshen-containing serum on the proliferation of HSCs were detected by MTT, the expression of SuFu mRNA and DYRK2 mRNA were detected by RT-PCR, the expression of SuFu and DYRK2 proteins were tested by Western blot. Compared with blank control group, the expression of DYRK2 mRNA and DYRK2 proteins were enhanced obviously after stimulated the HSCs of rats by leptin (P < 0.01), but the expression of SuFu mRNA and SuFu proteins were decreased significantly (P < 0.01). Compared with the model group, after cyclopamine group (Hh pathway inhibitor), Danshen-containing serum and colchicine were interfered, the expression of DYRK2 mRNA and DYRK2 proteins were decreased clearly (P < 0.01), but the expression of SuFu mRNA and SuFu proteins were increased significantly (P < 0.01 or P < 0.05). Compared with model group, adding purmorphamine (Hh pathway agonist) to model group and making it activate could increase the expression of DYRK2 mRNA and DYRK2 proteins, but the expression of SuFu mRNA and SuFu proteins were decreased significantly (P < 0.01). Compared with the model group, using the Danshen-containing serum to interfere the purmorphamine group could make the expression of DYRK2 mRNA and DYRK2 proteins decrease and the expression of SuFu mRNA and SuFu proteins increase significantly (P < 0.01). Danshen-containing serum would inhibition the activation and increment of HSCs by interfering the expression of SuFu and DYRK2 which were induced by leptin.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatic Stellate Cells ; drug effects ; metabolism ; Humans ; Liver Cirrhosis ; drug therapy ; genetics ; metabolism ; Male ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Repressor Proteins ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry

China ; Congresses as Topic ; Fatty Liver ; Humans

Objective To observe the therapeutic effects of the co-administration of γ-aminobutyric acid (CABA), sodium dimercaptopropane sulfonate (Na-DMPS) and vitamin B6 in large doses on liver and heart of rats with acute tetramine intoxication, and compare their separate effects of either GABA or Na-DMPS alone with those of the triad combination. Method Thirty rats were randomized into control group (n = 6), tetramine intoxication without treatment group (n = 6), tetramine intoxication treated with GABA group (n = 6), tetramine intoxication treated with Na-DMPS group (n = 6) and tetramine intoxication treated with triad combination (GABA + Na-DMPS + vitamin B6, GNDV n = 6) group. Samples of blood, liver tissue and heart tissue of rats with acute tetramine intoxication were collected immediately two hours after medication with different drugs. Serum alanine aminotrasferase (ALT), aspartate aminotransferase (AST), creatine kinase (CK) and creatine kinase isoenzyme (CK-MB) were measured, and the pathological changes of liver tissue and heart tissue were observed under microscope. Results The symptoms of poisoning were apparently relieved and the latency for convulsion/muscular twitch were obviously delayed in poisoned rats treated with GABA, Na-DMPS and GNDV separately. Furthermore, combination group showed the latent period delayed longer than either GABA or Na-DMPS groups The GABA, Na-DMPS and GNDV significantly lowered the serum levels of ALT, AST, CK and CK-MB in rats with tetramine intoxication, and those serum levels of enzymes were lower in GNDV group than those in either GABA group or Na-DMPS group. However, there were no difference in those serum enzymes between GABA group and Na-DMPS group. Moreover, the intoxicated rats treated with combination treatment had the slightest pathological changes in liver and heart (GNDV < GABA or Na-DMPS). Conclusions The co-administration of γ-aminobutyric acid, sodium demercaptopropane sulfonate and vitamin B6 in large doses for the treatment of tetramine intoxication is a method of choice.

OBJECTIVETo evaluate the antitumour efficacy of shRNA plasmid specifically targeting Jab1 gene.

METHODSThe nude mouse tumor model was made by subcutaneous injection of human laryngeal carcinoma Hep-2 cells. The tumor growth was monitored after intratumoral injection of pJab1, pKB plasmids and saline. Jab1 and p27 expressions in tumour tissues were examined by immunohistochemistry staining and RT-PCR.

RESULTSMean volume of the pJab1-treated tumors was (267.60 ± 88.19) mm(3), significantly less than that of tumors treated with pKB plasmids (832.20 ± 140.39) mm(3) or saline (895.40 ± 145.93) mm(3) (F = 36.73, P < 0.001). Immunohistochemistry showed that the expression of Jab1 protein was significantly reduced in the pJab1-treated group (32.40% ± 5.59%) compared to the control groups, whereas the expression rate of p27 protein in the pJab1 group (76.80% ± 6.30%) was significantly increased compared to the control groups (P < 0.001). The down regulation of Jab1 protein by pJab1 plasmid was consistent with mRNA expression confirmed by RT-PCR. The level of Jab1 mRNA level in the pJab1-treated group (0.65 ± 0.03) was significantly lower than the control groups (F = 558.00, P < 0.001), however, p27 mRNA, was 0.80 ± 0.02, had no significant alteration (F = 1.52,P > 0.05).

CONCLUSIONSThe pJab1 plasmid results in downregulation of Jab1 in an xenograft tumour model of human laryngeal carcinoma Hep-2 cells and significantly inhibits the tumour growth in vivo. This suggests that pJab1 plasmid specifically targeting Jab1 gene expression could be an effective therapy for human laryngeal carcinoma.

Animals ; COP9 Signalosome Complex ; Carcinoma, Squamous Cell ; genetics ; Cell Line, Tumor ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Laryngeal Neoplasms ; genetics ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Peptide Hydrolases ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection ; Xenograft Model Antitumor Assays

Objective To construct the objective system of 5+3 integrated clinical medicine personnel training mode.Methods For the attitude,knowledge and ability clinical medical students should have,who received 5+3 integrated training,we adopted literature research,expert interviews intending to make a preliminary questionnaire items,and invited experts and graduates to modify the expressions of the items a number of times,and eventually formed the final questionnaire that met the study objective.We asked 500 teachers in basic medicine,clinical medicine,and medical humanities and so on,to evaluate the importance of the questionnaire.406 valid questionnaires were recovered and the effective recovery rate was 81.2％.Epidata 3.1 and SPSS 19.0 were used for survey data summary statistics,exploratory factor analysis and factor weighting method were used to get the goal of talent training base,and on these basis target system was constructed.Results In 5+3 integrated medical education model,clinical medical personnel training target system included knowledge,attitude and ability of the part,a total of 9 target groups.Each group contained elements in the training target system for different weights,among which attitude included a common factor,proportion of 27.51％;knowledge consisted of three factors,the proportion of 29.34％;and the ability contained five common factors,accounting for highest proportion of 43.15％.Conclusion 5+3 integrated clinical medical talents training target system established in this study highlights the ability requirements,emphasizes the comprehensive quality training,which accords with the principles of talent training goal and the requirements of 5+3 medical education model,has certain guiding significance for the curriculum reform,and can be used for reference in the development of talents training program for medical colleges and universities.

Objective To investigate the effect of long-term high-fat diet on insulin resistance and the morphology and function of islets in rats and the relationship between them.Methods Thirty normal male Wistar rats (8 weeks old) were divided into two groups and fed either with normal chow (NC,n=15),or high-caloric and high-fat diet (HF,n=15).Insulin resistance was assessed by euglycemic hyperinsulinemic clamp technique. The insulin secretory function of islets was evaluated by intravenous insulin releasing test.Morphological and quantitative analysis of pancreatic tissues was performed by double-label insulin and glucagon immunohistochemistry.Proinsulin mRNA was detected by RT-PCR.Results The glucose infusion rate (GIR) in HF rats was significandy lower than that in NC rats [(5.83?0.79)mg?kg~(-1)?min~(-1) vs (7.60?1.29)mg?kg~(-1)?min~(-1),P＜0.05].Immunohistochemistry showed that HF rats had larger islet size [(15168?1327)?m~2 vs (6264?1840)?m~2,P＜0.01] and significantly reduced insulin relative concentration of?cells[(-5.15?0.03) vs (-4.81?0.17),P＜0.01],as compared with NC rats.The islet relative?cell volume was decreased signifieandy (P＜0.01),whereas the relative?cell volume was increased (P＜0.01).So the ratio of?to?were lower in HF [(4.68?1.01) vs (11.84?3.82),P＜0.05].The peak of insulin secretion in intravenous insulin releasing test in HF was at 10 min,whereas that in NC rats was at 5 min.AUC (area under curve) 10-60 rain of insulin in HF was higher than that in NC rats [(152.51?34.53)mIU?L~(-1)?min~(-1) vs (86.40?21.21) mIU?L~(-1)?min~(-1),P＜0.01].There was no difference in proinsulin mRNA levels between two groups. Conclusion Long-term high-caloric and high-fat diet results in early impairment of islet morphology and function, as well as significant insulin resistance,suggesting that the compensation ability of islets has already been impaired in the early course of type 2 diabetes mellitus.

Objective To investigate the effect and mechanism of transplantation of marrow multi- function stem ceils in treating femoral head necrosis.Methods Sixty japanese rabbits were divided into A,B and C groups randomly.After creation of the models of hormone induced necrosis of femoral head;A group was designated as the treatment,B as the control and C as the normal groups.The bone marrow of A group was extracted and isolated and then injected into the left femoral head and the right femoral head was decompressed by drilling only.The rabbits were killed at 8 weeks after the treatment and changes in various parameters were observed,including imaging data of molybdenum target films,CT and MRI;routine pathology with HE staining and ultrastructural alteration by election microscopy.Results Eight weeks after the treatment of transplantation of marrow multifunction stem cells,the X-ray showed only a little change but the typical appearances were revealed by CT and MRI.Pathohistologic manifestation demonstrated decrease of empty bone lacuna,increase of osteoblast and new bone formation.Election microscopy displayed abundant organelles in osteoblasts with few empty bone lacuna,in addition the tansplantation of marrow multifunction stem cells could obtain better reconstraction for the involved femoral head.Conclusions The treatment of transplantation of marrow muhifunction stem cells in femoral head necrosis could accelerate the process of repairing,worthy to be acknowledged as a good and valuable management in rabbits.(J Intervent Radial,2007,16:122-126)