A series of researches concerning the relationship between multidrug transporters and drug resistance in medically intractable epilepsy have been done. There is accumulating evidence demonstrating that P-glycoprotein (PGP) is a candidate to cause AEDs resistance. The effect of PGP inhibitor-verapamil on the intracellular AEDs accumulation in a MDR(multidrug resistant) K562 was investigated. The multidrug resistant (overexpression of PGP) cell line K562/Dox was established and the intracellular PHT and CBZ accumulation in multidrug resistant cell line and non- multidrug resistant cell line was observed. After PGP inhibitor-verapamil was applied to the two cell lines, the concentration change of PHT and CBZ in MDR cell was observed. The results were found: compared with non-MDR cell line K562, which IC50 was significantly increased in MDR cell line K562/Dox after PHT and CBZ was applied; verapamil could decrease significantly the level of IC50 in MDR cell line K562/Dox, and the reversal index were 2.5 and 1.5. The concentration of PHT and CBZ in MDR cell line K562/Dox was lower than that in non-MDR cell line K562, and verapamil significantly increased the concentration of PHT and CBZ in MDR cell line K562/Dox(P

Objective To study the expressions of Chemerin in hepatocellular carcinoma (HCC) and HCC cell lines,and to demonstrate the relationship between the expressions and prognosis.Methods The expressions of Chemerin protein in normal hepatocellular tissues,HCC and their paired tissues were detected by immunohistochemical staining of SABC; the expressions of Chemerin protein in HCC,cell lines and immortalized hepatocyte cell lines were detected by RT-PCR.Results The positive expression rates of Chemerin were 56.67％,90.00％,100％ in HCC,HCC cell line and normal hepatocellular tissues respectively,and the differences were significant (P＜0.05).The expressions of Chemerin mRNA in HCC paired tissues were higher than in HCC (P＜0.05).There was a higher expression of Chemerin mRNA in immortalized hepatocyte cell line LO2 ; but a lower expression in HCC cell lines.The expressions of Chemerin were related to lymph node metastasis,portal vein tumour thrombi,differentiation and TNM stage but not related to sex,age,tumour size,HBsAg and AFP.Conclusions Down-regulated expression of Chemerin may play an important role in the development,progression and metastasis of HCC.It may be a molecular marker for prognosis of HCC.

BACKGROUND:Bone marrow mesenchymal stem cellcontent is less under normal conditions, and easily confounded with other cells. Therefore, to establish a simple feasible in vitro cultured amplification method and to obtain a large number of stable bone marrow mesenchymal stem cells are of important theoretical significance and application value. OBJECTIVE:To establish a simple feasible in vitro culture and amplification method and to obtain numerous stable bone marrow mesenchymal stem cells. METHODS:Bone marrow mesenchymal stem cells were isolated from Sprague-Dawley rats and cultured in vitro using density gradient centrifugation and adherence separation screening. cellmorphological changes were observed under an inverted light microscope. Submicroscopic structure of cells was observed under a transmission electron microscope. Living cellnumber was counted using Trypan blue exclusion method. cellgrowth curve was drawn. cellcycle was analyzed using flow cytometry. The expression of c-kit and CD45 was measured using immunocytochemistry. CD45 expression was analyzed using flow cytometry. RESULTS AND CONCLUSION:(1) celladhesion and pseudopodia could be seen under an inverted light microscope at 24 hours after inoculation;cellcolony formed at 4 days;90%cellconfluence was found at 14 days. After passage, the cells were tended to homogenous and became fibroblast-like cells, which showed whirl-like growing or flamboyancy growing. (2) The size of passage 3 bone marrow mesenchymal stem cells was smal and nucleolus was large, with clear nucleoli under a transmission electron microscope, with the presence of sparse chromatin and low electron density. There was microvil us on the surface of cells, abundant ribosomes could be seen in the cytoplasm with few other cellular organs such as endoplasmic reticulum, mitochondria and Golgi complex. These showed that ultrastructural structure had undifferentiated features. (3) The number of passage 3 bone marrow mesenchymal stem cells was reduced at 1 day after inoculation. The cells began to grow at 2 days, and entered the period of exponential growth at 3 days, entered the flat period at 7 days and the number of cells began to decrease at 9 days. Growth curve exhibited“S”shape. (4) The percentage of S phase cells was 21.1%as detected by flow cytometry after passage 3 bone marrow mesenchymal stem cells were stained by DNA. (5) Immunocytochemistry and flow cytometry had proved that the c-kit positive rate of cells was 53.3%and CD45 positive rate was 1.68%. (6) After osteogenic induction of mesenchymal stem cells for 16 days, cells became oval, and short processes connected each other. The cytoplasm was dark, suggesting that it may be rich in rough endoplasmic reticulum and Golgi complex. They can secrete osteoid. Results verified that obtained bone marrow mesenchymal stem cells stably grew, and actively proliferated.

Objective To study the value of 4‐item examination of gastric function in the evaluation of disease severity and course of peptic ulcer .Methods The 4‐item examination of gastric function ,including PGⅠ ,PGⅡ ,PGⅠ /PGⅡ ,gastrin 17 and He‐licobacter pylori(Hp) positive rate ,were detected by adopting enzyme linked immunosorbent assay (ELISA) in 175 cases of peptic ulcer(study group) and 60 persons undergoing physical examination(control group) .The Spearman correlation coefficient was used to analyze the relationship between the 4‐item gastric function examination with the severity and course of peptic ulcer .Results The levels of PGⅠ ,PGⅡ ,gastrin 17 and Hp positive rate in the study group were significantly higher than those in the control group(P<0 .05) ,while PGⅠ /PGⅡwas significantly lower than that in the control group(P<0 .05) .The values of PG Ⅰ /PGⅡfrom high to low were in turn the scarring stage group ,healing stage group and active stage group ,other indexes from high to low were in turn active stage group ,healing stage group and scarring stage group .The levels of PGⅠ ,PGⅡand gastrin 17 and Hp posi‐tive rate in the patients with disease course ≥3 years were significantly higher than those in the patients with disease course<3 years(P<0 .05) .The severity and disease course in the patients with peptic ulcer were positively correlated with the levels of PGⅠ ,PGⅡ ,gastrin 17 and Hp positive rate(P<0 .05) and negatively correlated with PG Ⅰ /PGⅡ(P<0 .05) .Conclusion In the pa‐tients with peptic ulcer ,4‐item gastric function examination is closely correlated with the severity and course of illness ,which can serve as the index for evaluate the severity and course of disease ,thus increases the clinical treatment pertinence ,reduces the occur‐rence of antibacterial drugs abuse ,and is worthy of being promoted and applied in clinic .

Objective:To study the expression of CD137L in laryngeal carcinoma, and to analyze its clinicopathological significance.Method:The expression of CD137L in 50 laryngeal carcinoma specimens and 9 normal laryngeal mucous tissues were detected by immunohistochemical staining.Result:The positive CD137L staining were found in all 50 cases of laryngeal carcinomas (100%), while its staining were negative in normal laryngeal mucous. There was significant difference between two groups(P<0.01). The positive ratio of CD137L staining had no relationship with the factors such as age, sex and tumor site, while it had significant correlation with the pathological stage,T stage and lymph node metastasis.Conclusion:The expression of CD137L might play an important role in the development of laryngeal carcinomas.

Vectors used to carry foreign genes play an important role in gene therapy, among which, the adeno-associated virus (AAV) has many advantages, such as nonpathogenicity, low immunogenicity, stable and long-term expression and multiple-tissue-type infection, etc. These advantages have made AAV one of the most potential vectors in gene therapy, and widely used in many clinical researches, for example, Parkinson's disease. This paper introduces the biological characteristics of AAV and the latest research progress of AAV carrying neurotrophic factor, dopamine synthesis related enzymes and glutamic acid decarboxylase gene in the gene therapy of Parkinson's disease.

Objective To investigate the level of homocysteinemia in type 2 diabetes mellitus with neuropathy,and observe the effect of medical intervention of hyperhomocysteinemia on diabetic neuropathy.Methods 200 patients with type 2 diabetes mellitus plus neuropathy and 200 control were collected.Patients with diabetic neuropathy plus hyperhomocysteinemia were divided into two groups:medical intervention group(15mg/d folic acid,20mg/d vitamin B6)and control group.The level of homocysteinemia in plasma was measured,and neuropathy score was evaluated by clinical and electrobiological methods before and three months after treatment.Results The morbidity of hyper-homocysteinemia in type 2 diabetes mellitus with neuropathy was 40%,and the levels of homocysteinemia was higher in treatment group than in control(P<0.05).Compared with the control,the levels of homocysteinemia in group of medical intervention was lower(P<0.01),and neuropathy score and nerve conductive velocity showed an obviously improvement after 3-month-treatment(P<0.05).Conclusions Hyperhomocysteinemia is an independent risk factor for diabetic neuropathy in humans.Efficiently intervening the levels of homocysteinemia could improve the electrobiological activity and neuropathy score.

Objective To study the effect of P-glycoprotein (PGP) in regulation of penetration of carbamazepine and phenytoin through blood-brain barrier, as to indicating the entry of epileptic drugs into the brain restricted by PGP and deducing the multidrug resistance mechanisms of refractory epilepsy. Methods The microdialysis probe was placed into the cortex of rats, and after systemic injection of carbamazepine and phenytoin, dialysate was collected and the antiepiletic drug concentration in the extracellular fluid of the cerebral cortex was determined by high-performance liquid chromatography (HPLC). Then observing whether the concentration of drugs in extracellular fluid can be enhanced by PGP’s inhibitor verapamil. Results Verapamil significantly enhanced the concentration of carbamazepine in extracellular fluid of the cerebral cortex during 60—90 minutes after injection ((1.74?0.28)?g/ml in 60 minutes and (1.87?0.31)?g/ml in 90 minutes,P

Objective To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from human gastric carcinoma and paired normal gastric mucosa tissues and to analyse the differential expression proteins between the two types of the tissues. Methods The total proteins of gastric carcinoma and paired normal gastric mucosa tissues were seperated by immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE).After silver staining, the differential expression proteins were analyzed by using Imaging Master 2D analysis software. Results ⑴Average protein spots were 1049?67, 1097?73 in gastric carcinoma and paired normal gastric mucosa respectively,the matched spots were 835?48, 953?56 in both the two types of tissues respectively, and the average matching rate was 79.6%, 86.7% respectively. ⑵A total of 769?45 protein spots were matched between the electrophoretic maps of 5 gastric carcinoma and paired normal gastric mucosa tissues. The 81 differential protein spots were identified between the two types of the tissues. 17 protein spots were expressed only in the gastric carcinoma and 24 protein spots were expressed only in the normal gastric mucosa. The expression levels of 26 protein spots in the gastric carcinoma were higher than those in the normal gastric mucosa, and the expression levels of 14 protein spots in the gastric carcinoma were lower than those in the normal gastric mucosa. Conclusion In this study, the well-resolved,reproducible 2-DE patterns of gastric carcinoma and paired normal gastric mucosa tissues were established and some differential proteins between the two types of tissues were found. These data will be helpful to further screen specific biomarkers of gastric carcinoma.

Objective To investgate the role of aquaporin-4(AQP4) in secondary cerebral edema after ischemia/reperfusion injury in rats.Methods When the models of reversible middle cerebral artery occlusion were established, the alterations of cerebral edema and BBB were evaluated by measuring water and Eval's Blue (EB) contents of cerebral tissue, and the expression of AQP4 in brain was observed by Western Blot at different time point after reperfusion. At last, the correlation between expression of AQP4 and water and EB contents of cerebral tissue were analysed.Results There were found that water and EB contents of cerebral tissue in rat models significantly higher than those of control group at different time point after ischemia/reperfusion ( P