Objective To identify the distribution feature of methylenetetrahydrofolate reductase (M T HFR) gene polymorphism of Buyi ,Dong ,Miao nationality in Guizhou .Methods The MTHFR(677 and 1 298) genotypes of Buyi ,Miao and Dong healthy indi-viduals were determined by TaqMan-MGB probe genotyping method and constructed haplotypes .Results There were significant difference of MTHFR 677C/T genotype and allele frequencies among 3 groups(P<0 .05) ,There was significant difference of geno-type between Buyi and Miao nationality ,and there were significant differences of genotype frequencies in Buyi nationality and Dong and Miao nationality(P<0 .01) .There were no differences of MTHFR 1298A/C genotype frequencies among Buyi ,Dong and Miao nationality(P> 0 .05) .Buyi nationality had the lowest frequency in double wild homozygous type (677CC/1298AA) ,677TT/1298CC double mutation homozygous and 677TT/1298AC combination in above three minorities was not found .There were linkage disequilibrium between 677C/T and 1298A/C in Buyi and Miao nationality .Conclusion The genotypes frequencies of MTHFR 677T T/1298AC are significant differences among different regions and different ethnic groups .

Constructing prokaryotic expression vector pKY-RMBAY by gene recombination and research its optimizing productive conditions.By PCR technology synthesizing the gene of the RMBAY with preference codon of E.coli and the RMBAY gene was inserted into high efficiency expression vector pKYB-MCS.Expressed fusion proteins in E.coli ER2566 were purified with Chitin-Beads column.Fusion proteins binding on Chitin-Beads was cut on N-terminus of intein due to the induction of ?-mercaptoethanol and the target peptide RMBAY was released.The RMBAY was identified by mass spectrum.Experiment results showed RMBAY can be high efficiently expressed in E.coli ER2566,with optimizing productive conditions the yield of the RMBAY may be 6.7mg/L fermentation product and its purity is greater than 98%.The molecular weight of RMBAY is 3.887 kDa by mass spectrum and that accords with its theory value.

Objective To investigate the mechanism of acute inflammatory response and tissue repair when rats accepted transplanted bone marrow mesenchymal stem cells (MSC) in severe acute pancreatitis (SAP).Methods A total of 40 rats were randomly divided into 4 groups which included the normal group (n=10),the model severe acute pancreatitis group (n=10),the transplanted bone marrow mesenchymal stem cells group (n =10),and the combination of bone marrow mesenchymal stem cells and granulocyte colony-stimulating factor (G-CSF) group (n=10).To cure the acute severe pancreatic injury caused by SAP,rats were injected with EDU-labeled MSCs and granulocyte colonystimulating factor (Gr-CSF).The conversion rate of MSCs to pancreatic cells or MSCs to endothelial cells was detected to assess the role of MSCs in tissue regeneration and repair.The expression of amylase,interleukin-6 (IL-6),and interleukin-10 (IL-10) in serum was detected to assess the role of MSCs in an acute inflammatory response.Results The concentrations of amylase and IL-6 were reduced and the concentration of IL-10 was increased in MSC and MSC+G-CSF groups after the onset of SAP.Flow cytometry showed a small amount of MSCs converting to endothelial or pancreatic cells,but the conversion rate was relatively low.Conclusions MSCs can play an important role in the antipre-release of inflammatory mediators,reducing acute immune response to control the acute inflammatory response in SAP.Moreover,MSCs can repair a damaged pancreas by converting into both pancreatic and endothelial cells.

We wished to select a cold-adapted genotype G1P[8] ZTR-68 rotavirus (China southwest strain) in MA104 cells for possible use as a live vaccine. ZTR-68 was recovered originally from children with diarrhea. The virus was cultivated at 37 degrees C at the first passage. Then, the cultivation temperature was decreased stepwise by 3 degrees C per eight passages. In total, the virus was passaged 32 times, and cultivation was terminated at 28 degrees C. Biological characteristics of the virus were analyzed during serial passages. There was no difference between the migration patterns of genomic dsRNA segments according to polyacrylamide gel electrophoresis of original and cold-adapted viruses. Infectious and red cell-agglutination titers of cold-adapted virus were lower than those of the parent virus. Also, the virus formed small-size plaques with irregular shapes at 31 degrees C and 28 degrees C. These results suggested that a genetically stable attenuated virus can be obtained through serial cold-adapted passages. Thus, an alternative strategy is provided by cold-adaption for development of attenuated live rotavirus vaccines.
Adaptation, Physiological ; China ; Cold Temperature ; Diarrhea ; virology ; Female ; Genotype ; Humans ; Infant ; Male ; Rotavirus ; genetics ; growth & development ; isolation & purification ; physiology ; Serial Passage ; Virus Cultivation ; Virus Replication

Animals ; Chlorine ; toxicity ; Interleukin-8 ; metabolism ; Lung ; drug effects ; metabolism ; pathology ; Male ; Pulmonary Emphysema ; chemically induced ; metabolism ; Rats ; Rats, Wistar ; Time Factors ; Tumor Necrosis Factor-alpha ; metabolism

Objective To analyze CMV DNA stability of 30 EDTA plasma samples in the order of magnitude between 300 and 100 000 copies/mL over a 21 day period. Methods Thirty plasma samples were grouped into three categories according to the CMV DNA loads , including low CMV DNA contents , intermediate CMV DNA loads and high CMV DNA loads. Ten milliliters of whole blood was freshly collected from each patient. Plasma samples without hemolysis were divided into 1-ml aliquots. One aliquot was processed immediately (Day 0) for baseline PCR assays. The remaining aliquots were then processed after one , two, three, seven, 14 or 21 day of storage at 4℃. Results There was no significant difference between the mean of the difference time point in viral loads following storage at 4 ℃ by paired-samples t test, including Day 1 compared to Day 0 (t = 1.654, P =0.109), Day 2 compared to Day 0 (t = 1.487, P = 0.148), Day 3 compared to Day 0 (t = 1.609, P = 0.118), Day 7 compared to Day 0 (t=0.831, P=0.413), Day 14 compared to Day 0 (t=1.721, P=0.096), and Day 21 compared to Day 0 (t=0.244, P=0.810). Conclusion The concentration of CMV DNA in all samples stored at 4 ℃ for 21 days did not differ significantly from the baseline viral load ,and it was not observed the trend in continued degradation in different time point (Day 1, 2, 3, 7 and 14).

Objective To explore the effect of excessive fluoride on expression of mRNA and protein of Wnt3a and β-catenin in rats' osteoblasts and its correlation with pathogenic mechanism of fluorosis.Methods Thirty-six healthy SD rats,weighting 100-120 g and according to body mass,were randomly divided into three groups(twelve in each group).The rats of control were fed wich tap water(fluoride ＜ 1 mg/L) and the experimental rats were exposed to NaF(low-fluoride group:5 mg/L,high-fluoride group:50 mg/L) added to the drinking water to establish the chronic fluorosis model.After fed for eight morth,all rats were killed and metaphysic of femoral was collected.Rat dental fluorosis was observed and bone fluorine was detected by ashing-fluorin ion selective electrode method.The content of bone alkaline phosphatase (BALP) and tartrate-resistant acid phosphatase 5b(TRACP 5b) in rats' serum was detected by enzyme-linked immunosorbent assay (ELISA).The morphologic changes of the bone were observed by microscopy.The expression of mRNA and protein of Wnt3a and β-catenin in osteoblasts of rats was analyzed with gray scale by hybridization in situ and immunohistochemistry methods,respectively.Results Detection rate of dental fluorosis,fluoride contents of urine and bone were significantly increased [control group:0.0％,(1.26 + 0.17)mg/L,(305.58 ± 91.26)mg/kg; low-fluoride group:66.7％,(2.06 ± 0.64)mg/L,(632.33 ±123.21)mg/kg; high-fluoride group:91.7％,(7.69 ± 1.96)mg/L,(1088.75 ± 156.16) mg/kg] in the rats treated with fluoride,the difference between groups was statistically significant(χ2 =21.6; F =36.57,467.02; all P ＜0.05).The contents of BALP and TRACP-5b in rats' serum were significantly different between groups(F =89.57,7.68; all P ＜ 0.05).Compared with control group[(16.24 + 1.57)U/L],the contents of BALP in rats' serum of the low-fluoride and high-fluoride groups[(31.47 ± 5.30) and (54.61 ± 2.27)U/L] were increased gradually(all P ＜0.05).Compared with the low-fluoride group,the value in the high-fluoride group decreased significantly (P ＜ 0.05).The contents of TRACP-5b in rats' serum of low-fluoride group[(3.45 ± 1.85)U/L] were elevated significantly(all P ＜ 0.05) compared with the control group[(1.26 ± 0.23)U/L] and the high-fluoride group[(2.74 ± 1.85)U/L].The bone cortices were thickened and the bone trabecula was broadened,arranged closely together in chronic fluorosis rats with significant difference compared with the control group.In the low-fluoride and high-fluoride groups,the expression levels of Wnt3a and β-catenin mRNA (low-fluoride group:132.87 ± 5.72 and 132.57 ± 9.56; highfluoride group:135.60 ± 6.64 and 137.87 ± 9.16) were markedly elevated with significant difference,respectively (F =12.47,5.96; all P ＜ 0.05) compared with those in control groups(119.86 ± 5.04 and 120.58 ± 7.84) by hybridization in situ(P ＜ 0.05),but there was no statistical significance (P ＞ 0.05) of the level of Wnt3a and β-catenin mRNA between low-fluoride and high-fluoride groups.In the low-fluoride and high-fluoride groups,the protein expression of Wnt3a and β-catenin (low-fluoride group:137.50 ± 4.32 and 140.85 + 3.54; high-fluoride group:142.65 ± 11.84 and 152.52 ± 4.64) were markedly elevated with significant difference,respectively (F =10.07,53.82; all P ＜ 0.05) compared with those in control group (124.01 ± 2.63 and 126.75 ± 4.65) by immunohistochemistry(all P＜ 0.05),Wnt3a protein production in the low-fluoride group was increased without statistical significance compared with the high-fluoride group (P ＞ 0.05).But the protein production of β-catenin in the lowfluoride group was elevated with significant difference compared with the high-fluoride group(P ＜ 0.05).The mRNA and protein production of Wnt3a were positively correlated with the mRNA and protein production of β-catenin (r =0.731,0.658; all P ＜ 0.05).Conclusions Rat bone tissue lesions caused by excessive fluoride may be associated with an increased expression of Wnt3a and β-catenin mRNA and protein in osteoblasts.In chronic fluorosis,fluoride stimulates the overexpression of Wnt3a and β-catenin in the Wnt signal transduction pathway,enhances bone osteogenesis and causes skeletal fluorosis.

It was hypothesized that the VPAC1 agonist may exert anti-obesity functions because VPAC1 is involved in the anorexigenic effects and the anti-inflammatory function of pituitary adenylate cyclase-activating polypeptide (PACAP)/vasoactive intestinal polypeptide (VIP). Furthermore, our in vitro test showed that the expression of VPAC1 increased significantly after the 3T3-L1 adipocytes were differentiated, and that incubation of adipocytes with VPAC1 agonist (10-1 000 nmol/L per 1x10(6) cells) resulted in stimulation of lipolysis. To test the effect of VPAC1 agonist [Lys15, Arg16, Leu27]-VIP (1-7) GRF (8-27) on diet-induced obesity (DIO), we further designed the following two in vivo experiments: (1) Mice were fed on high-fat diet (HFD) and intraperitoneally (i.p.) treated with VPAC1 agonist simultaneously for 28 d; (2) Mice were given HFD for 35 d, and subsequently fed on the same HFD and i.p. treated with VPAC1 agonist for the next 28 d. The physiological indices, including body weight, weight of white adipose tissue, plasma glucose and blood lipid, were collected. The results showed that treatment with VPAC1 agonist inhibited ingestion significantly and prevented the elevations in body weight and the weights of the white adipose tissues (epididymal and dorsal) induced by HFD. The increases in plasma glucose, cholesterol, triglycerides and LDL induced by HFD were also down-regulated in mice treated with VPAC1 agonist. VPAC1 agonist treatment also improved the glucose tolerance. Therefore, VPAC1 agonist treatment inhibits the development of the obesity induced by HFD and helps to improve the morbidities associated with DIO.
3T3-L1 Cells ; Adipocytes ; drug effects ; Animals ; Body Weight ; Diet, High-Fat ; Mice ; Obesity ; drug therapy ; Receptors, Vasoactive Intestinal Polypeptide, Type I ; agonists ; Vasoactive Intestinal Peptide ; pharmacology

OBJECTIVETo develop a transparent, non-toxic, non-irritating anti-fogging agent with long-lasting effect for nasal endoscopy.

METHODSThe anti-fogging agent was prepared by mixing ethanol, propylene glycol, polyoxyethylene lauryl ether, sodium dodecyl sulfate, polyethylene glycol 400 and deionized water at different proportions based on an orthogonal test design. Twenty-seven test samples of the anti-fogging agents were obtained, which were colorless, transparent, and non-irritating, with a pH value of 7-8. Storz00 nasal endoscopy and its imaging system were used to test the anti-fogging time of the 27 samples, and each agent was tested for 3 times with medical Seoul iodine and 95% ethanol as control.

RESULTSThe optimal composition of the anti-fogging agent was 20% ethanol, 10% propylene glycol, 20% polyoxyethylene lauryl ether, 4% sodium dodecyl sulfate, 4% polyethylene glycol, 42% deionized water. The anti-fogging time of this agent reached 15 min, significantly longer than that of medical Seoul iodine (4 min) and 95% ethanol (18 s).

CONCLUSIONThis anti-fogging agent for nasal endoscopes is colorless and safe and has a long anti-fogging time by forming a homogenous transparent membrane over the endoscopic lens.

Endoscopes ; Endoscopy ; methods ; Ethanol ; Nose ; surgery ; Polyethylene Glycols ; Sodium Dodecyl Sulfate ; Solutions ; chemistry

According to the trend of the development of the quality education, it is necessary to re-form the traditional homework assignments in medical microbiology for improving the innovation ability of students. This program attempts to reform the homework on clinical medicine undergraduates. First, the teacher must master diversified knowledge, and then the student design and finish the homework combine with personal characteristic from the training requirements of experimental confirmation, theoretical cognition and social survey. The questionnaire shows the personalized homework can help students to master the pro-fessional knowledge, stimulate study interest and improve the comprehensive quality.