Objective: To investigate the role of uncoupling protein -2 in development of nonalcoholic fatty liver diseases ( NAFLD) ,and explore the effect of glutathione ( GSH) on NAFLD. Methods: 32 SD rats were randomly divided into basic diet - treated group ( B group) ,model group( M group) , Pathology group( P group) and GSH treated group( G group). The following parameters were determined by biochemical analysis; superoxide dismutase( SOD) , glutathione(GSH) , malondialdehyde (MDA). Liver histopathologic e-valuation was performed by hematoxylin - eosin staining. Serum tumor necrosis factor alpha(TNF?) level was done by enzyme -linked immunoadsordent assay ( ELISA). Ucp2 and TNFaproteins expression in hepatocytes was examined by immunohistochemistry, and UCP2mRNA expression by semi - quantitative reverse transcription - polymerase chain reaction( RT - PCR). Results: ( 1 ) compared with B group, no evident increase in liver MDA,SOD and GSH of M group rats as well as serum TNF?level were observed. Neither marked change in number of hepatocytes with TNF?positive was shown by immunohistochemistry staining, but an increase in UCP2 mRNA and UCP2 protein expression was seen in M group rats. (2) Compared with M group, a significant increase in serum TNF? and MDA levels was found in P group .while a significant decrease in liver homogenate SOD and GSH was observed . An apparent increase in number of hepatocytes with TNF?and UCP2 positive was shown by immunohistochemistry staining, as well as UCP2mRNA level by RT-PCR. (3) Compared with P group, a significant decrease in serum MDA and a significant increase in liver homogenate SOD, GSH levels was found in G group. A little lower level of UCP2 expression was shown in G group than in P group. Conclusion; High level of serum and tissue TNFaand UCP2 over - expression are concomitant with an extreme decrease even depletion in the liver ATP in NAFLED model rats, that cause oxidative stress and lipid super - oxidation in liver, and induce or exacerbate fatty liver disease progression to steatohepatitis even fibrosis. Glutathione seems to be effective to some extent for NAFLD by protection of liver function through replenishment of exogenous antioxidants.

Objective To investigate changes in indicators related to inflammatory cytokines, coagulation and fibrinolysis system in patients with disseminated intravascular coagulation (DIC) and multiple organ dysfunction syndrome (MODS) resulted from sepsis. Methods In total, 97 patients diagnosed as sepsis were divided into different groups and their plasma concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), tissue factor (TF) and tissue factor pathway inhibitor (TFPI), tissue plasminogen activator (t-PA), and plasminogen activator inhibitor 1 (PAI-1) were measured by ELISA and plasma activity of protein C (PC:A) was measured by chromogenic substrate assay. Results Plasma concentrations of TNF-α and I L-6 were (38 ± 12) ng/L and (77 ± 9) ng/L, respectively, in patients of sepsis complicated with DIC, much higher than those without DIC [ ( 17±6) ng/L and (45 ± 6), respectively], P <0.05;and (63±25) ng/L and (103±28) ng/L, respectively, in those complicated with MODS, significantly higher than in those without MODS [ (29 ± 7 ) ng/L and (48±9)ng/L, respectively], P < 0/05 and those without DIC [( 17 ± 6) ng/L and (45 ± 6) ng/L, respectively], P <0. 05;as well as significantly higher in the dead than in survivors (P <0. 05). Plasma activity of protein C was (32 ± 10) percent in those with DIC and (24 ± 12) percent in those with MODS, both significantly lower than in those without DIC [ (57±28) percent] and without MODS [ (55 ± 17) percent], respectively, P <0. 05, as well as significantly lower in the dead than in survivors, P <0. 05. Plasma concentrations of t-PA and PAI-1 were significantly higher in sepsis patients with DIC [(48±17)μg/L] than that in those without DIC [(103 ± 38)μg/L], P < 0.05. Conclusions Inflammatory cytokines play important roles in development of DIC as well as MODS in patients with sepsis. Decreased activity of protein C and increased plasma level of PAI-1 can result in deposition of fibrin on the vessel wall and thrombosis, which can be used as indicators of poor prognosis for patients with DIC.

Objective To compare the therapeutic effects of cordyceps sinesis (CS) and reduced glutathione on experimental non-alcoholic fatty liver disease (NAFLD) rats and explore the possible molecular mechanisms. Methods After NAFLD rats were induced by high-fat diet and were treated by CS and reduced glutathione. The histopathologic changes of livers were evaluated. The levels of TG and FFA in serum and liver were measured. The levels of SOD and ATP in liver were measured too. Results (1)In the development of NAFLD, extensive adipose degeneration, inflammatory cell infiltration and necrosis, local fibrous tissue hyperplasia were found in the liver. The increase of TG, FFA in the serum and liver and decrease of SOD and ATP in the liver were seen. (2)In CS treated group, adipose degeneration had been alleviated with slightly inflammatory cells infiltration and no necrosis or fibrosis had been found. The concentrations of TG and FFA were decreased in the serum and liver, but SOD and ATP increased. (3)In glutathione treated group, adipose degeneration of liver and inflammatory cells infiltration remained obviously with focus or punctiform necrosis, but without fibrosis. The increase of SOD in liver was distinguished. No changes of TG, FFA, UCP-2 and ATP had been detected. Conclusion Both CS and reduced glutathione have therapeutic effects on NAFLD, by preventing the generation of liver fibrosis. CS has a better therapeutic effects on metabolic disturbance or accumulation of lipid and energy metabolic imbalance of liver cells.

Objective To investigate the difference in complete blood count (CBC) within the defined time among 28 hospitals with different grade in Urumchi City, explore its influencing factors,offer the countermeasures, and further to provide scientific evidence for the mutual recognition among different medical institutions. Methods Totally 28 laboratories with different grade were enrolled in the investigation. Three fresh blood samples were employed to perform complete blood counting, in-cluding normal and pathological specimens. The measuring time and turn around time (TAT) were de-fined. Results According to the literature report on biological variability of testing results, the per-missible deviation of CBC was tentatively prescribed as follows: WBC 15% ;RBC 6% ;HB 7% ;HCT 6 % ;PLT 25 %. Qualification decision was based on the external quality assessment (EQA) criteria of Clinical Laboratory Centre of the Health Ministry. Qualification: the total score of five measured items is higher than or equal to 80%;disqualification: the total score of five measured items is lower than 80%. For the first and second sample, 26 laboratories (92.86%) were qualified;for the third sample (only 21 laboratories enrolled), 14 laboratories (67%) were qualified. Conclusion 28 laboratories par-ticipated in this test. The results of five items are congruent for the first and second sample (normalspecimens) in 28 laboratories, and the measured results are comparable. By contrast, the comparative result is unsatisfactory for the third sample (pathological specimen), mainly due to the narrow linear range of equipment. Meanwhile, the laboratory quality management should be strengthened.

Objective To study the characteristics of molecular genetics concerning Chinese mitochondr ial encephalomyopathy,lactic acidosis and stroke-like episodes (MELAS). Methods A3243G and T3271C point mutations in the mtDNA of muscle and (or) blood cells were investigated in 9 patients with MELAS and some o f their maternal relatives from 7 families by using PCR-RFLP. Furthermore,mut ant mtDNA in the sample harboring mutation was quantitatively analysed. Results The mtDN A A3243G point mutation was unanimously identified in tissues of all patients an d 1 of their relatives. However,the T3271C point mutation was identi f ied in none of series in our study. The proportion of mtDNA A3243G was 46.8%～ 61. 0% in muscle (4 cases) as well as 26.8%～50.3% in blood (9 cases). In the 3 p atie nts with muscles and blood cells available,their mutant mtDNA proportion in mus cle is consistently higher than in blood cells. The study of leukocytes of some maternal relatives from 6 families showed that,while only 1 proband had a siste r harboring A3243G mutation and none of the mothers of another 3 probands or sib lings of the other 2 probands had the point mutation. However,the sons of 2 pr o bands had not only phenotype of MELAS,but also mtDNA A3243G point mutation in t heir blood. Conclusion mtDNA A3243G mutation highly exists in the series with M E LAS syndrome in our study and can be detected in various tissues,which is consi stent with reports abroad. However,most of our cases are sporadic rather than m aternal inherited. It is presumedly caused by a de novo mutation. Whether i t is due to ethnic difference or sporadic event needs to be investigated further.

Objective:To explore the effect of glycynhizinatis on serum IgE level of rat model and patients with asthma.Methods:Trie rats were divided into four groups randomly: normal control, asthmatic model, model treated with GL, model treated with DS. The rats were sensitized and challenged with ovalbumin to establish an asthmatic model. The established models were measured by breathing curves, pathological changes in pulmonary tissue slices and the level of serum IgE. Furthermore, serum IgE level of patients with asthma was also detected. Results: High amplitude of breathing curve and fast rhythm were showed in group of asthmatic model, there were more inflammatary cells infiltration in the tiny bronchial wall, hyperplasia of cup-shaped cells and thicker smooth muscle in the pulmonary tissue slices. Compared with normal control, the level of serum IgE in model group increased obviously. The rats treated with glycyrrhizinatis showed low amplitude of breathing curve, decreasing inflammatary cells infiltration in pulmonary tissue slices of tiny bronchial wall, no hyperplasia of cup-shaped cells, and decreased serum IgE level. There were no significant difference between the groups of GL and DS. At the same time, the serum IgE level in patients with asthma was observed. The results showed that the level of serum IgE was higher in patients with asthma than that in normal control (P

0.05). The median duration of survival was 11.5 months for TEP gr oup versus 8.5 months for EP group (P0.05,no statistical difference). The main toxicity wa s myelosuppression for the two groups.The incidence rate of Ⅲ～Ⅳ degree neutro p enia and thrombocytopenia was higher in the TEP group than that in the EP group( P

Objective To explore the intervention effect of emodin on organophosphorus poisoning induced respiratory failure.Methods 60 male Wistar rats of clean grade were randomly divided into:normal control group, model control group, positive drug group and emodin group, with 15 rats in each group.Except the normal control group rats were given intraperitoneal anesthesia, the right common carotid artery intubation, when rats stayed awake began a septic model.Blood gas analysis and serum level of oxygen free radicals and respiratory rate were compared before poisoning, respiratory failure, intervention of 5, 10, 30 min.Results Mouth breathing, slow respiratory frequency and cyanosis, appeared after exposure.Respiratory frequency decreased after exposure , compared with the positive drug group, respiratory frequency of emodin group 10 min and 30 min was higher ( P<0.05), PaO2, SaO2, BE decreased, PaCO2 increased after respiratory failure, Compared with the positive drug group, PaO2, PaCO2, SaO2 and BE of emodin group for the treatment of 10 min, 30 min was higher,(P <0.05).The level of oxygen free radicals in rats of each group had no significant difference before the exposure and the respiratory failure.Compared with the positive drug group, SOD and MDA of emodin group in 30 min after intervention were higher,( P<0.05 ) .Conclusion Emodin can improve the respiratory frequency of organic phosphorus poisoning induced respiratory failure ,improve blood gas analysis of the indicators and the level of oxygen free radicals.

Objective To observe the effect of capsaicin on lipid deposition in liver cells and the expression of autophagy,to provide new ideas and targets for the study of fatty liver disease.Methods The model of nonalcoholic fatty liver disease (NAFLD) was established by inducing LGF cell line of liver primary cells with oleic acid (40 μg/mL).The experiment was divided into blank control group,oleic acid group,capsaicin group[oleic acid--capsaicin(100 μmol/L)].The level of intracellular lipid was detected by oil red O staining and triglyceride (TG) kit.Western bolt was used to detect the expression level of autophagy marker molecule p62 and LC3,and to calculate the ratio of LC3 Ⅱ/LC3-Ⅰ.Results After treated with 40 μg/mL oleic acid for 24 h,the oil red O staining showed that orange-red lipid droplets were found in the oleic acid group,but there was no obvious orange-red lipid droplet formation in the blank group,which suggesting that the NAFLD cell model was established successfully by using 40 μg/mL oleic acid in vitro.Oil red O staining was observed,the cells in the capsaicin group were significantly less than the oleic acid group,and the content of triglyceride in the liver cells was significantly lower than that in the oleic acid group(P＜0.05).Western bolt test results showed that capsaicin group p62 protein levels were significantly lower compared with the oleic acid group(P＜0.05),while the LC3-Ⅱ/LC3-Ⅰ ratio was higher than that of oleic acid group(P＜0.05).Conclusion NAFLD model was established by inactivating LO2 cell line with oleic acid in vitro.Capsaicin stimulated NAFLD cells,upregulated the autophagy level of NAFLD cells and reduced intracellular lipid deposition,but the specific mechanism need further study.

ObjectiveTo construct human embryonic kidney cells (HEK293) modified with human preproenkephalin (hPPE) gene.MethodshPPE gene fragments were obtained from recombinant plasmid pcDNA3.1( + )/hPPE by using restriction endonuelease Hind Ⅲ and Not Ⅰ.Homologous recombination of lentivirus and hPPE gene was produced by using recombinant DNA technology.HEK293 cells were then transfected with the recombinant lentivirus vectors.The expression of hPPE gene in HEK293 cells was detected by Western blot.ResultsThe results of DNA sequencing indicated that the positive clone of recombinant lentivirus was completely consistent with sequencing of hPPE in Genebank.The titer of the concentrated virus was 2.07 × 108 TU/ml.GFP fluorescence was not seen in HEK293 cells transfected with the lentiviral vector under fluorescence microscope.A strong fluorescence was seen in HEK293 cells transfected with Ubc-GFP-L.V.empty viral vector.Positive expression of hPPE was demonstrated in HEK293 cells transfected with lentiviral vector by Western blot.Conclusion HEK293 cells modified with hPPE gene were successfully constructed and the target gene hPPE was stably expressed in HEK293 cells.