Objective To investigate the association between vascular risk factors and the location of intracranial artery stenosis (anterior versus posterior).Methods Magnetic resonance angiography(MRA) were examined in 374 acute stroke patients.It was divided into two groups (anterior and posterior intracranial artery stenosis group).Analyzed possible risk factors.Results Univariate analysis showed there were differences between anterior and posterior intracranial artery stenosis in systolic blood pressure,history of smoking,drinking and stroke status,and national institutes of health stroke scale (NIHSS) score at discharge,short-term prognosis,serum creatinine,triglyceride,low density lipoprotein cholesterol (LDL-C) (P ＜ 0.05).In multivariate logistic regression analysis,high blood sugar (OR =1.135,95％ CI:1.003-1.284),history of stroke(OR =1.133,95％ CI:1.007-1.276),good short-term prognosis (OR =5.987,95％ CI:1.441-24.873) were preferentially related to anterior intracranial artery stenosis,whereas history of smoking (OR =0.003,95 ％ CI:0.000-0.376),high serum creatinine values (OR =0.509,95 ％ CI:0.328-0.790),high triglyceride values (OR =0.054,95％ CI:0.004-0.645) and high LDL-C values (OR =0.096,95％ CI:0.015-0.608) were preferentially related to posterior intracranial artery stenosis.Conclusion Vascular risk factors appeared to exert different effects of risk for anterior and posterior intracranial artery stenosis.

0.05). The rate of RCC after HHSSCT was 4.7% in study group; after second NS soaking, the rate of RCC was 19.1% in control group. The difference between the two groups was statistically significant (P

Objective To establish a sandwich ELISA for early detection of HIV antigens using a mixture of monoclonal antibodies (McAb). Methods The ascites McAbs (anti-HIV-1 p24, anti-HIV-1 gp41, anti-HIV-1 gp120 and anti-HIV-2 gp36) were purified by the SAS and the affinity chromatography,and then were labeled with HRP by sodium metaperiodate. The establishing of sandwich ELISA for detecting the single HIV antigen and the tests of specificity and sensitivity of these systems were performed in advance.A proper ratio mixture of four screened McAbs was used as the capture antibody and a proper ratio mixture of four labeled antibodies was used as the detecting antibody. The method of using sandwich ELISA to detect HIV antigens was set up with these McAbs. Results The sensitivity of this method detecting HIV antigens are:0.625 pg/ml HIV-1 p24, 6.25 ng/ml HIV-I gp41,6.25 ng/ml HIV-I gp120 and 9.25 ng/mi HIV-2 gp36 in mixed HIV antigens. Conclusion The method of using several McAbs mixture in sandwich ELISA detecting HIV antigens was established an excellent sensitivity, which provides a novel idea for early detec-ting the HIV antigen.

Carbon-limited continuous culture was used to study the relationship between the growth of Aspergillus niger and the production of glucoamylase. The result showed that when the specific growth rate was lower than 0.068 h(-1), the production of glucoamylase was growth-associated, when the specific growth rate was higher than 0.068 h(-1), the production of glucoamylase was not growth-associated. Based on the result of continuous culture, the Monod dynamics model of glucose consumption of A. niger was constructed, Combining Herbert-Pirt equation of glucose and oxygen consumption with Luedeking-Piret equation of enzyme production, the black-box model of Aspergillus niger for enzyme production was established. The exponential fed-batch culture was designed to control the specific growth rate at 0.05 h(-1) by using this model and the highest yield for glucoamylase production by A. niger reached 0.127 g glucoamylase/g glucose. The black-box model constructed in this study successfully described the glucoamylase production by A. niger and the result of the model fitted the measured value well. The black-box model could guide the design and optimization of glucoamylase production by A. niger.
Aspergillus niger ; metabolism ; Batch Cell Culture Techniques ; Carbon ; Culture Media ; Glucan 1,4-alpha-Glucosidase ; biosynthesis ; Glucose ; Industrial Microbiology ; methods ; Oxygen

Objective To investigate the relationship between serum homocysteine ( Hcy ) and cognitive impairment in patients with cerebral infarction combined chronic obstructive pulmonary disease (COPD) after stroke, and observe the plasma Hcy levels and cognitive function improvement when treated with folic acid and vitamin B12 .Methods 87 acute cerebral infarction combined with COPD patients as the research object, then the general clinical data were recorded, hematology indexes ( Hcy, folic acid, vitamin B12 ) were determined, and their cognitive function with a simple mental state scale (MMSE) was assayed.According to the plasma Hcy levels, the subjects were divided into Hcy-normal group (n =21) and Hcy-increased group (n=66), then compare the cognitive function between the two groups.Hcy-increased subjects were randomly divided into intervention group (conventional treatment +folic acid 2.5 mg +VitB12 0.5 mg) and control group (conventional treatment).After six months follow-up, we retested plasma Hcy levels and MMSE assessment, comparison of plasma Hcy concentration change and cognitive function improvement between two groups.Results Compared with Hcy-normal group, plasma folic acid, VitB12 levels significantly decreased in Hcy-increased group (P<0.05).And Hcy concentration was negatively correlated with folic acid(r=-0.351,P =0.000)and VitB12(r=-0.242,P=0.015)levels.In addition, the MMSE, directional force and delayed recall score decreased in Hcy-increased group compared with the Hcy-normal group ( P<0.05 ).Hcy levels were significantly lower than the baseline level (P<0.05), MMSE and the sub-project of cognitive function score increased after treated with folic acid and VitB12 for six months, although there was no statistically significant difference.Conclusion Plasma Hcy level is associated with cognitive impairment in patients with cerebral infarction combined chronic obstructive pulmonary disease (COPD), patients treated with folic acid and VitB12 may help slow the recent cognitive dysfunction after stroke in the near future.

BACKGROUNDTo investigate the expression of COX-2 and its relation to clinical pathophysiological features and prognosis in non-small cell lung cancer (NSCLC).

METHODSThe expression of COX-2 protein was detected in 52 NSCLC tissues by immunohistochemical (S-P) method.

RESULTSThe positive COX-2 expression was observed in 25 (48.1%) cases of NSCLC tissues. The positive rate of COX-2 expression was 76.5% and 34.3% in adenocarcinoma and squamous cell carcinoma respectively (P < 0.01). The positive rate of COX-2 expression in T3+T4 disease (92.3%) was remarkably higher than that in stage T1+T2 (33.3%) (P < 0.01). There was a remarkable difference in COX-2 expression rate between clinical stage I+II (28.1%) and clinical stage III+IV (80.0%) groups (P < 0.01). The positive rate of COX-2 expression was 83.3% in those with lymph node metastasis, but only 17.9% in those without lymph node metastasis (P < 0.01). In addition, there were significant differences in positive rate of COX-2 expression among patients with ≤2, > 2 but < 5, ≥5 years of survival span respectively (P < 0.01).

CONCLUSIONSOverexpression of COX-2 in NSCLC, especially in adenocarcinoma, is closely related to invasion, lymph node metastasis and clinical stage of lung cancer. It may play a role in development of NSCLC, and also may be a prognostic marker.

Aim To explore the effect of chemical hypoxia-mimetic agent,cobalt chloride(CoCl2)on inflammatory reaction in human keratinocytes(HaCat cells).Methods After HaCat cells were treated with CoCl2 at different concentrations to set up a chemical hypoxia-induced cell model of skin injury,cell viability,intracellular reactive oxygen species(ROS),mitochondrial membrane potential(MMP),the levels of both interleukin 6(IL-6)and interleukin 8(IL-8)as well as the expression of heme oxygenase-1(HO-1)were detected.Results The viability of HaCat cells was reduced by CoCl2 at the concentrations from 500 to 3 000 ?mol?L-1,and the higher CoCl2 doses,the lower cell viability was.CoCl2 induced oxidative stress reaction(increasing ROS production and decreasing MMP).CoCl2 induced inflammatory reaction,enhancing the release of IL-6 and IL-8.CoCl2 at concentrations from 1 000 to 3 000 ?mol?L-1 upregulated HO-1 expression in HaCat cells.Conclusion CoCl2 induces not only oxidative stress,but also inflammatory reaction,increasing the release of both IL-6 and IL-8,as well as HO-1 expression.

BACKGROUNDTumor-necrosis factor related to apoptosis inducing ligand protein(TRAIL), like tumor-necrosis factor (TNF) and Fas, is a member of TNF cytokine supper family. Many researches have showed that TNF-α can reverse the resistance to some chemotherapeutic agents in cancer cell lines, and some anticancer drugs can result in up-regulations of death receptor (DR) and further lead to the enhancement of apoptosis induced by TRAIL. In order to clarify if TRAIL can reverse the resistance to cisplatin in cancer cells, the effects of recombinant human tumor-necrosis factor related to apoptosis inducing ligand protein (rhTRAIL) on apoptosis in human lung adenocarcinoma cell lines resistant to cisplatin (DDP) in vitro was explored.

METHODSHuman lung adenocarcinoma cell lines resistant to cisplatin, A549/DDP cells, were cultured in regular condition. At 24 hours after TRAIL and DDP, alone or combined, microculture tetrazolium (MTT) dye was used to evaluate the cytotoxic effects. And besides, to detect the apoptotic effects of rhTRAIL on A549/DDP cells, flow cytometry assay was used to test the apoptosis proportion, diphenylamine assay (DPA) was applied to detect the percent of DNA fragmentation and Caspase-3 chluorometric assay was performed to test the activity of Caspase-3 among these cells.

RESULTSA549/DDP cells were not sensitive to low-dose rhTRAIL alone. The rate of growth inhibition and the apoptotic indexes such as the apoptosis proportion, the percent of DNA fragmentation and the activity of Caspase-3, had all no significant changes with rhTRAIL concentration less than 25μg/L (P > 0.05). But treated with higher-dose rhTRAIL more than 50μg/L, the four values changed obviously: 68.6%, (27.13± 0.66)%, (37.4±2.0)% and 0.117±0.011, respectively (P < 0.05). With combination of different concentration of rhTRAIL and 3mg/L DDP, the cyto-toxic and apoptotic effect was comparatively more apparent. The combination of rhTRAIL and 3mg/L DDP presented synergistic effect on A549/DDP, 12.5μg/L concentration of rhTRAIL together with 3mg/L DDP could kill 30.4% of A549/DDP cells. Furthermore, the rate of cell apoptosis, percent of DNA fragmentation and activity of caspase-3 increased to (19.39±0.54)%,(17.3±4.1)% and 0.138±0.009, which were significantly different from those of rhTRAIL alone (P < 0.01).

CONCLUSIONSHigh-dose rhTRAIL can also induce the cells resistant to cisplatin to apoptosis, but the cytotoxic and apoptotic effects of rhTRAIL alone are weaker than those of combination of rhTRAIL and low-dose cisplatin which can augment the apoptotic effect induced by rhTRAIL. rhTRAIL is expected to be an efficient biologic drug for treatment of lung cancer resistant to chemotherapy.

Objective To explore whether the phosphorylation of NF-κB P65 subunit is involved in the cytotoxicity to and inflammation in an immortal human keratinocyte cell line HaCaT during cobalt chloride (CoCl2-induced chemical hypoxia. Methods HaCaT cells were treated with CoCl2 of 2 mmol/L to set up a chemical hypoxia-induced cell model of injury. Then, RNA interference was used to down-regulate the expression of P65 in CoCl2-induced HaCaT cells. After additional culture, cell viability was tested by cell counting kit8 (CCK-8), the levels of interleukin 6 (IL-6) and interleukin 8 (IL-8) were detected by ELISA kits, phosphorylated and total P65 protein was measured by Western blot. Results The exposure of HaCaT cells to 2 mmol/L CoCl2 for 0 to 4 hours enhanced the phosphorylation of P65, which began at 0.5 hour, peaked at 1.5 hours, and restored to the normal level at 4 hours, and the level of P65 phosphorylation was about 6.6 times that in the untreated control group. The CoCl2 of 2 mmol/L decreased the cell viability of HaCaT cells in a time dependent manner, and a significant difference was observed in the viability of HaCaT cells between CoCl2-treated and untreated HaCaT cells at 2, 4, and 6 hours (P ＜ 0.05, 0.01, 0.01 ). The release of IL-6 and IL-8 from HaCaT cells was also promoted by CoCl2 treatment. The knockdown of P65 expression with siRNA markedly suppressed the CoCl2-induced cytotoxicity to and increase in the release of IL-6 and IL-8 from HaCaT cells,despite of an increment in cell viability by about 11%. Conclusion The phosphorylated P65 subunit mediates CoCl2-induced cytotoxicity and inflammatory injury to HaCaT cells.

OBJECTIVETo research the effect of Jianpi Jiedu decoction (JPJDT) to PTEN/ERK1 of athymic mice with hepatocellular carcinoma.

METHODN2 male BALB/c athymic mice models were built by Bel-7402 with an indirect method. After 24 h of postoperation, the 90 athymic mice were distributed randomly into JPJDT groups: A, B, C, D, E, F, G, NS, FT each group had 10 athymic mice. Another 10 male BALB/c athymic mice without HCC was treated by NS as normal control (DZ). Group A to G were treated by intragastric administration with JPJDT that had been deliquated into 7 kinds of density for 8 wk. Group NS were were treated by intragastric administration with Sodium Chloride for 8 wk. Group FT were were treated by intragastric administration with FT207 (tegafur) for 8 wk . At last, athymic mice were sacrificed. PTEN/ERK1 was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry (PowerVision two-step histostaining reagent).

RESULTThe expression intensity of PTEN: The result showed that the intensity of PTEN in the normal hepatic tissue was the highest, and then latero-cancer tissue, the lowest was cancer tissue. In the normal hepatic tissue, the intensity of PTEN in Group B, D, E was higher than the Group NS, Group FT, Group DZ (P < 0.05). In the latero-cancer tissue, the intensity of PTEN in Group D was higher than the Group NS (P < 0.05). In the cancer tissue, the intensity of PTEN in Group JPJDT was higher than the Group NS and Group FT (P < 0.05). The expression intensity of ERK1: The result showed that the intensity of PTEN in the cancer tissue was the highest, and then latero-cancer tissue, the lowest was normal hepatic tissue. In the latero-cancer tissue, the intensity of ERK1 in Group FT was higher than the Group NS and Group JPJDT (P < 0.05). In the cancer tissue, the intensity of PTEN in Group NS and Group FT was higher than the Group C, D, E, G, F (P < 0.05). The correlation between PTEN and ERK1: The result showed that there was inverse correlation between the expression intensity of PTEN and ERK1 in the cancer tissue (P < 0.01).

CONCLUSIONOne of mechanism of antitumous effect of JPJDT maybe up-regulate anti-oncogene PTEN, restrain the signal way of ERK1, suppress the proliferation of hepatoma carcinoma cell. The carcinogenesis of primary hepatic carcinoma may exist the deletion of PTEN. Owing to low expression or deletion of PTEN in the cancer tissue, ERK1 signal transduction pathway cannot be actively suppressed which was activated by carcinogenic factor. So hepatoma carcinoma cell multiplicated.

Animals ; Carcinoma, Hepatocellular ; genetics ; metabolism ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Liver Neoplasms ; genetics ; metabolism ; Male ; Mice ; Mice, Nude ; Mitogen-Activated Protein Kinase 3 ; metabolism ; PTEN Phosphohydrolase ; metabolism