50 000 bp)was extracted from frozen gastric cancer tissue and their corresponding normal samples and used for RLGS assay.The genome DNA was digested by methylation-sensitive restriction enzyme Not Ⅰ, and labeled by radioisotope ~(32)p,then separated by two-dimensional electrophoresis and autoradiography. Experimental conditions for each step were optimized step by step.DNA fragment sequences for the dots on scanning profile were identified according to the human RLGS database.Results RLGS assay was set up and used to get the RLGS profiles of the representative tested samples successfully.These profiles can display more than 1 200 available dots averagely,the profiles of high quality DNA sample can display more than 1 800 dots which is the average level at an excellent RLGS lab,discrepant dots which had weaken or enhanced signals and their sequence information were obtained.The result can be reproduced.Conclusion The RLGS assay is established,stabilized for detection of DNA methylation of tissue samples.

Objective To investigate the regulative effect of the extract of Radix Ranunculi Ternati,Radix Sophorae Flavescenti,Prunella vulgaris L.and Stellera chamaejasme L.on cellular immunity induced by multiple drugs resistant bacillus tuberculosis(MDR-TB) from pneumoconiosis patients complicated with tuberculosis in rats.Methods MDR-TB model in rats was induced by MDR-TB.Normal control group were feed by standard feed,model group were irrigated by normal saline,and the other groups were respectively feed by the extract of Chinese herbal medicines.The content of IFN-γ,IL-4,IL-10 and IL-12 were examined by ELISA.RT-PCR was used to detect the mRNA levels of them.Results The content of IFN-γof the four extracts of Chinese herbal medicines were (2.01 ±0.73 ),( 1.92±0.56),( 1.98 ±0.67 ) and (1.94±0.59) pg/ml,IL-4 were (6.01±1.46),(6.12±1.35),(6.47±1.46) and (6.15±1.44) pg/ml,IL-10 were (12.09±3.07),( 12.45±4.01 ),( 12.13±3.43) and (12.54±3.78) pg/ml,IL-12 were (2.99±0.89),(2.75±0.84),(3.02±0.86) and (2.89±0.75) pg/ml.Compared to the model group,they resulted in significant in serum IFN-γ,IL-12,IL-4 and IL-10 (P＜0.05 or P＜0.01 ),the mRNA levels of them were significantly (P＜0.05 or P＜0.01 ).Conclusion The four extracts of Chinese herbal medicines can enhance the cellar immunological function in rats from up-regulation of the level of genetic transcription,accordingly provide the theory base of healing of pneumoconiosis patients complicated with tuberculosis with them.

Objective To improve the scientific literature novelty assessment by studying the characteristics and novelty of researches in clinical medicine.Methods The objects, levels, locations, methods and novelty of 380 scientific literature novelty assessment files in Library of Guilin Medical College from 2012 to 2014 were analyzed by chi-square test.Results The novelty of researches was mainly manifested as an effect index in 2012-2013 and as an in-tervention factor in 2014(x2=110.12, P<0.01).The novelty of effect index was mainly manifested as a compre-hensive index in 2012-2013 and as a sensitivity index in 2014(x2=44.10, P<0.01).Conclusion Understanding and keeping abreast of the characteristics and novelty of researches in clinical medicine can serve them better and more effectively encourage their innovation .

Objective To analyze the affection and clinical significance of CD26/DPP4 on CD4+T cells and its cytokines in patients withCrytococcalMeningitis.Methods Peripheral blood was collected from 36 patients diagnosed withCrytococcal Meningitis in Changhai Hospital and Changzheng Hospital,Shanghai from August,2011 to December,2015,meanwhile 36 health controls’was also acquired.Peripheral blood mononuclear cell (PBMC)was separated by density gradient centrifuga-tion,CD26+CD4+T and CD26-CD4+T cell groups were classified by Flow Cytometry,the expression level of cytokines was tested by reverse transcriptase-polymerase chain reaction (RT-PCR).The correlation between DPP4 activity,CD26+CD4+T (%)and APACHE II score,IL-17,TNF-α,IL-4,IFN-γwas measured by Pearson coefficient.Results CD26+CD4+T(%)between experimental and control groups was 13.35±3.83 vs 8.39±2.14 (t=6.78,P<0.000 1).DPP4 activity was 50.89±17.21 mU/ml vs 73.83±20.24 mU/ml (t=5.18,P<0.000 1),with statistically significant differences.In ex-perimental groups,CD26+CD4+T (%)was positively related with APACHE II score,IL-17,TNF-α(r=0.431,0.564, 0.688,P=0.003 8,0.001,0.004 6).DPP4 activity was negatively interrelated with APACHE II score,IL-17,TNF-α,IFN-γ(r=-0.544,-0.489,-0.678,-0.734;P<0.001).Conclusion CD26/DPP4 may be involved in the pathogenesis of Crytococcal Meningitis through regulation of Th subgroups,and it was the potential therapeutic target and the predicted marker of the disease.

Objective To observe the effects of comprehensive measures of changing grain,selenium supplementation,off-site education and resettlement on prevention of children's Kashin-Beck disease in Aba state.Methods Fifty eight villages in Aba Kashin-Beck disease areas were chosen as intervention points in Aba state Sichuan province from 2007 to 2011.Based on the implementation of prevention and control measures,the villages were divided into off-site education + changing grain + selenium supplementation group and resettlement + off-site education + changing grain + selenium supplementation group,Geletuo town of Seda county,Ganzi state was selected as a control point,and right-hand anteroposterior X-ray examination(including the wrist) was carried out on children aged 6-13 from 2007 to 2011 annually.Clinical and X-ray diagnosis of Kashin-Beck disease was made in accordance with the Diagnostic Criteria of Kaschin-Beck Disease(GB 16003-1995).The effects of prevention and control measures were evaluated by comparing the child X-ray detection rate before and after the implementation of the measures.Results The average X-ray positive detectable rate of children in the intervention points was 2.07％(66/3181),2.72％ (69/2540),1.16％ (35/3017),0.56％ (19/3397) and 0.56％ (24/4273),respectively from 2007 to 2011,with a downward trend (x2trend =66.74,P ＜ 0.01).There was a downward trend in the average X-ray positive detectable rate of children in off-site education + changing grain + selenium supplementation group [1.60％(29/1809),2.63％ (39/1484),1.29％ (25/1941),0.64％ (15/2332),0.42％ (10/2379)] and resettlement + off-site education + changing grain + selenium supplementation group [2.70％ (37/1372),2.84％ (30/1056),0.93％(10/1076),0.38％ (4/1065),0.74％(14/1894)] (x2trend=30.97,35.19,all P ＜ 0.01).The average X-ray positive detectable rate of children in the intervention group was 0 from 2007 to 2010,and was 1.61％ (1/62) in 2011.The difference of X-ray positive detectable rate was not statistically significant in the control group in the 5 years from 2007 to 2011.The difference of children's X-ray positive detectable rate was not statistically significant between control group and intervention group.Conclusions The effect of taking changing grain,selenium supplementation,off-site education and resettlement comprehensive measures to prevent children's Kashin-Beck disease is not significant in those places where the state of Kaschin-Beck disease is not active.

· AIM:To explore the modulation effects of 17-β estradiol (E2) and tamoxifen (TAM) in chronic intraocular hypertension mouse model.· METHODS:We performed anterior chamber injection of magnetic beads to induced chronic ocular hypertension models.Adult C57BL/6 male mice were used in the experiments and randomly divided into four groups:control,Beads group,E2 group and E2+TAM group.The intraocular pressure (lOP) were measured by Tonolab tonometer.Central retinal thickness was evaluated by HE staining.Brn3a as a specific marker of retinal ganglion cells (RGCs),were stained and counted by immunohistochemistry (IHC) staining.Glial fibrillary acidic protein (GFAP) as a marker of proliferation of astrocytes,was quantified using western blotting.· RESULTS:The IOP level was significantly elevated after anterior chamber injection of magnetic beads compared to control group (P＜0.05),while in E2 and E2+TAM group,the IOP levels were reduced (P＜ 0.05 vs Beads group),especially in E2+TAM group in 2wk.The RGCs happened to degenerated in Beads group after 2wk,while the effects were reversed by E2+TAM (P＜0.05).The central retinal thickness showed no significant statistical difference among the four groups after 2wk (P ＞ 0.05).The expression level of GFAP increased caused via beads injection,however,decreased in E2 and E2+TAM group (P＜0.05).· CONCLUSION:E2 and E2+TAM could both effectively decrease the IOP level in chronic intraocular hypertension mouse model,increase the survival of RGCs from high intraocular pressure,suppress expression of GFAP,which indicated neuroprotective effects of E2 and TAM in glaucoma through an anti-inflammatory effects.

AIMTo study the chemical components from the bioactive extract of Herpetospermum pedunculosum, a Tibetan medicinal herb for liver diseases.

METHODSThe isolation and purification of this extract were conducted by means of silica gels column chromatography and preparative HPLC. The structures of the compounds were elucidated based on their physical and chemical features, and spectral data.

RESULTSTwo lignans were isolated from this extract. They were elucidated as herpetone (I), dehydrodiconiferyl alcohol (II).

CONCLUSIONCompound I is a new compound with the rare structure of sesqui-norlignan. Compound II is isolated from the genus of Herpetospermum for the first time.

Chromatography, Gel ; methods ; Chromatography, High Pressure Liquid ; Cucurbitaceae ; chemistry ; Lignans ; chemistry ; isolation & purification ; Molecular Structure ; Phenols ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Seeds ; chemistry ; Silica Gel ; Silicon Dioxide

Homo sapiens longevity assurance homologue 2 (LASS2) is a novel gene isolated from a human liver cDNA library by our laboratory, and it is a human homologue of the yeast longevity assurance gene LAG1 (Saccharomyces cerevisiae longevity assurance gene). According to our previous results, LASS2 could interact with subunit c of vacuolar type H(+)-ATPase (V-ATPase), and the overexpression of LASS2 could inhibit the cell growth of a human hepatocellular carcinoma (HCC) cell line, SMMC-7721. In order to understand the role of the interaction between LASS2 and V-ATPase in HCC cell growth, we transiently transfected plasmid pCMV-HA2-LASS2 into HCCLM3, a HCC cell line without the significant expression of endogenous LASS2. The pH-sensitive fluorescence probes, BCECF and BCECF-AM, were used to measure the intracellular and extracellular H(+) concentrations of HCCLM3 cells respectively. The effect of LASS2 gene on apoptosis was evaluated with Annexin-V/FITC and propidium iodide (PI) by flow cytometry. Western blot was used to detect cytochrome c (Cyt c) in the cytosol and mitochondria, as well as pro-caspase-3 in cytosol. The results showed that the cell growth of LASS2-transfected HCCLM3 cells was significantly inhibited compared with that of the mock control. LASS2 transfection increased intracellular H(+) concentration of HCCLM3 cells, while decreased extracellular H(+) concentration. Moreover, LASS2 transfection significantly enhanced the apoptosis of HCCLM3 cells. In LASS2-transfected cells, the amounts of Cyt c increased in the cytosol, while decreased in the mitochondria. Meanwhile, the expression of pro-caspase-3 in the cytosolic extracts was decreased. These results implicate that LASS2 gene might increase intracellular H(+) of HCC cells via the interaction with V-ATPase, thereby inducing cell apoptosis through mitochondrial pathway.
Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Liver Neoplasms ; pathology ; Membrane Proteins ; metabolism ; RNA, Small Interfering ; Sphingosine N-Acyltransferase ; metabolism ; Transfection ; Tumor Suppressor Proteins ; metabolism ; Vacuolar Proton-Translocating ATPases ; metabolism

OBJECTIVETo design a platform for microarray data analysis and processing in the browser/server mode running in Linux operating system.

METHODSBased on the Apache HTTP server, the platform, programmed with Perl language, integrated R language and Bioconductor packages for processing and analysis of the input data of oligonucleotide arrays and two-color spotted arrays. Users were allowed to submit data and parameter configurations to the platform via the web page, and the results of analysis were also returned via the web page.

RESULTSWith an easy operation and high performance, the platform fulfilled the functions of processing, quality assessment, biological annotation and statistical analysis of the data from oligonucleotide arrays and two-color spotted arrays. Using the platform, we analyzed the gene expression profiles in Mtb-stimulated macrophages of three clinical phenotypes, namely latent TB (LTB), pulmonary (PTB) and meningeal (TBM), and obtained valuable clues for identifying tuberculosis susceptibility genes. We also analyzed the effect of INH treatment on Mycobacterium tuberculosis gene expression in various dormancy models, such as hypoxia and KatG mutant, and found that a set of genes responded to INH treatment during exponential growth but not in dormancy, and that the overall number of differentially regulated genes was reduced in the cells in low metabolic state.

CONCLUSIONThe platform we have constructed integrates comprehensive resources, and with a user-friendly interface, allows direct result visualization to facilitate microarray data analysis.

Computational Biology ; methods ; Databases, Genetic ; Gene Expression Profiling ; methods ; Genetic Predisposition to Disease ; Humans ; Macrophages ; metabolism ; microbiology ; Mycobacterium tuberculosis ; genetics ; Oligonucleotide Array Sequence Analysis ; Software ; Tuberculosis ; genetics ; immunology ; User-Computer Interface

The biological characters of Sindbis virus strain of Yunnan(YN87448 strain) were studied by the test of the filtration, acid-resistant, ether-resistant, CPE, susceptibility of animal, HA, plague, determination of virus titres, and the cross-HI, cross-IFAT and PRNT as well.The results indicated that YN87448 strain belongs to Sindbis virus, Alphavirus, Togaviridae. YN87448 strain virus was plaque purified(PYN87448). The biological character of PYN87448 strain virus was studied too. PYN87448 strain virus will be used in the molecule biological test.