Objective To investigate the dynamic changes of serum levels of neuron-specific enolase(NSE),endothelin(ET)and calcitonin gene-related peptide(CGRP)and their clinical significance in the elderly patients with acute cerebral infarction.Methods One hundred and twenty elderly patients with acute cerebral infarction(ACI),60 elderly patients with lacunar infarct,60 elderly patients with hypertension and 60 elderly patients with cerebral artherosclerosis were enrolled. The areas of infarction were measured and the venous blood samples at different times were collected after cerebral infarction to determine the concentrations of NSE,ET and CGRP by radioimmunoassay. Results There were dynamic changes of the plasma levels of NSE,ET and CGRP.In the early time the plasma levels of NSE and ET of the elderly patients with acute cerebral infarction were significantly higher than those of the elderly patients with cerebral artherosclerosis,or with hypertension,or with lacunar infarct(P＜0.01,P＜0.01,P＜0.05,respectively),and were gradually declined along with timing.In ACI group,the level of NSE began to increase gradually after 24 hours,reached the highest in 2 days,and decreased to normal after 14 days,but the level of ET was always higher than those in the cerebral artherosclerosis and hypertension groups.The ET levels in lacunar infarct and hypertension groups were also significantly higher than in the cerebral artherosclerosis group(P＜0.01).However,when compared with the cerebral artherosclerosis and hypertension groups,the plasma concentrations of CGRP in cerebral infarction and lacunar infarct groups were obviously lower(P＜0.01),and increased gradually.We also found the larger the infarction area,the lower the level of CGRP.Conclusions The NSE,ET and CGRP concentrations are associated closely with acute cerebral infaction.Monitoring the level of NSE is applicable for early diagnosis of cerebral infarction.

OBJECTIVE:To establish a method for simultaneous determination of fatty acids in Isaria cicadae,Cordyceps militaris and C.sinensis,and to compare the difference of the contents of fatty acids among above medicinal herbs.METHODS:GC-MS method was adopted.Chromatographic condition:the determination was performed on TG-5MS gas phase capillary column with carrier gas of nitrogen at the flow rate of 1.2 mL/min.The inlet temperature was 290 ℃ by splitlesssampling;valve opening time was 1 min,and volume of sample was 1 μL.Mass spectrometry condition:the ion source is an electrospray ionization source.The temperatures of ion source and transmission line were 280 ℃,the initial temperature of the chromatographic column was 80 ℃ (gradient elution),ionization voltage was 70 eV.Solvent delay time was 5 min,and scan mass range was m/z 30-400.RESULTS:The linear ranges of tetradecanoic acid,pentadecanoic acid,palmitic acid,palmitoleic acid,heptadecanoic acid,heptadecenoic acid,docosahexaenoic acid,methyl oleate,linoleic acid,arachidonic acid,eicosenoic acid,diolefinic acid,eicosatrienoic acid,heneicosanoic acid,behenic acid,tricosanoic acid,lignoceric acid were 1.400-44.520 μg/mL(r=0.999 8),2.091-93.721 μg/mL(r=0.999 7),3.146-85.856 μg/mL(r=0.998 2),1.664-61.444 g/rnL(r=0.998 7),1.773-64.983 g/mL(r=0.999 5),1.781-68.421 μg/ mL (r=0.999 7),1.706-55.606 μg/mL (r=0.999 8),1.439-47.989 μg/mL (r=0.999 6),1.738-66.908 μg/mL (r=0.999 6),2.086-94.206 μg/mL(r=0.999 5),1.356-44.966 μg/mL(r=0.999 4),1.444-56.814 μg/mL(r=0.999 7),1.375-52.335 μg/mL(r =0.999 8),1.512-60.312 μg/mL(r=0.999 5),1.450-59.760 μg/mL(r=0.999 7),1.427-58.757 μg/mL(r=0.999 1),1.269-58.109 μg/ mL(r=0.999 3),respectively.The limit of quantitation was no more than 1 764.71 μg/mL,and the limit of detection was no more than 529.42 μg/mL.RSDs of precision,stability and reproducibility tests were all lower than 2.0％.The recoveries were 84.87％-108.93％ (RSD ranged 0.19％-2.23％,n=6).There were 17 fatty acids in C.militaris,16 fatty acids in Ⅰ.cicadae and 16 fatty acids in C.sinensis.The contents of unsaturated fatty acids in above medicinal herbs were higher than that of saturated fatty acids.The content of fatty acids in artificial cultivated Ⅰ.cicadae was mostly higher than other medicinal herbs.COCLUSIONS:The method is simple,accurate,stable and reproducible.It can be used for simultaneous determination of fatty acids in I.cicadae,C.militaris and C.sinensis.Above 3 kinds of medicinal materials.From the perspective of fatty acid content,the quality of artificial cultivated I.cicadae is best.

OBJECTIVETo investigate the effect of the selective PI3K inhibitor and MEK inhibitor on KRAS and PTEN co-mutated non-small cell lung cancer cell line NCI-H157 and the relevant mechanisms.

METHODSNCI-H157 was cultured routinely and treated with different concentrations of the two inhibitors. Cell proliferation was detected by MTT cell cycle assay. Based on the MTT results the cells were divided into four groups: the control group, PI3K inhibitor group (GDC-0941, 0.5 and 5.0 µmol/L), combination group I (0.5 µmol/L AZD6244 + 0.5 µmol/L GDC-0941) and combination group II (5.0 µmol/L AZD6244 + 5.0 µmol/L GDC-0941). Colony formation assay was performed to detect colony formation efficiency. The cell cycle and apoptosis were analyzed by flow cytometry. The expression of protein related to apoptosis was tested with Western blot.

RESULTSCell growth was inhibited by the two inhibitors. Combination groups led to stronger cell proliferation inhibition: combination group Ishowed synergistic effect of their actions and combination group II showed an additive effect; in both groups, there were decreased colony number [(77.2 ± 1.54)/well vs (61.50 ± 2.12)/well, P < 0.01] and [(51.00 ± 4.00)/ well vs (22.50 ± 3.53)/well, P < 0.01]; and enhanced apoptotic ratios [(18.30 ± 0.82)% vs (21.32 ± 0.56)%, P < 0.01] and [(27.14 ± 1.58)% vs (42.45 ± 4.42)%, P < 0.01]. In addition, compared to the PI3K inhibitor alone group, the NCI-H157 cells in the combination groups showed increased G0/G1 phase and decreased S phase (P < 0.01). Western blotting showed that the combination groups demonstrated significantly decreased expression of cyclin D1 and cyclin B1, increased p21 and cleaved PARP and decreased bcl-2/bax ratio, compared to the PI3K inhibitor only group.

CONCLUSIONThe combined inhibition of PI3K (AZD6244) and MEK (GDC-0941) has synergistic effects on the proliferation of NCI-H157 cells, but such effects appear to be in a dose-dependent manner.

Apoptosis ; drug effects ; Benzimidazoles ; administration & dosage ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin B1 ; metabolism ; Cyclin D1 ; metabolism ; Dose-Response Relationship, Drug ; Drug Synergism ; Humans ; Indazoles ; administration & dosage ; pharmacology ; Lung Neoplasms ; genetics ; pathology ; Mitogen-Activated Protein Kinase Kinases ; antagonists & inhibitors ; metabolism ; Mutation ; PTEN Phosphohydrolase ; genetics ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; metabolism ; Poly(ADP-ribose) Polymerases ; metabolism ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Proto-Oncogene Proteins p21(ras) ; metabolism ; Signal Transduction ; Sulfonamides ; administration & dosage ; pharmacology ; bcl-2-Associated X Protein ; metabolism ; ras Proteins ; genetics

OBJECTIVEEstablish the model of mouse with chronic hepatitis B virus (HBV) and nonalcoholic fatty liver disease (NAFLD).

METHODSTake 100 HBV transgenic, BALB/c mice of 4 weeks old, with each gender half. Then pick out 70 mice in one group to feed high-fat feed and the rest to feed normal feed. At the end of week 16, random kill 10 mice of high-fat, then liver tissue and serological detection target identification model is established in this paper. After that, divide the mice into model group and comparison group with 30 mice in each group. Feed model group with high-fat feed, comparison group with normal feed and normal group with normal feed till week 72 (including previous 16 weeks). Kill 10 mice of each group at the end of week 24, 48 and 72 respectively, fully automatic biochemical instrument detection of serum ALT, AST, TC, TG, FBG, fluorescence quantitative PCR method to detect HBV-DNA, chemiluminescence detection of HBsAg, liver biopsy after HE staining to evaluate histology change, observe mice model of dynamic evolution.

RESULTS(1) Feed high fat feed after 16 weeks, mice's weight, serum ALT, AST, TC, TG, FBG and blood biochemical indicators increased, HBV-DNA positive, liver HE staining obviously big blister fatty degeneration of liver cells and within the lobule lymphocytes infiltration, NAFLD activity score (NAS) getting close to NASH, the model of chronic HBV carries with NAFLD mouse built successfully. (2) The TC and TG values of model group in each period were higher than that of comparison group and normal group. (3) In week 24 and 72, HBV-DNA values of each group are obvious different from the other two groups and the difference can be applied to statistical significance (P < 0.05). (4) In week 48 and 72, NAS of each group are obvious different from the other two groups and the difference can be applied to statistical significance (P < 0.05).

CONCLUSIONS(1) Chronic HBV carries with NAFLD mice model can be established by HBV transgenic mice fed by high fat feed. (2) NAFLD accelerates the liver disease of the mice carrying HBV to some extent.

Animals ; Disease Models, Animal ; Fatty Liver ; complications ; pathology ; virology ; Female ; Hepatitis B virus ; genetics ; isolation & purification ; physiology ; Hepatitis B, Chronic ; complications ; pathology ; virology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Non-alcoholic Fatty Liver Disease

OBJECTIVE: To obtain the genetic polymorphism data of two STR loci D2S1399 and D5S2500 in Eastern Chinese Han population. METHODS: Blood samples or buccal swabs of unrelated Han individuals living in eastern China were analyzed using PCR-nature polyacrylamide gel electrophoresis-sliver staining method. RESULTS: 11 alleles of D2S1399 and 9 alleles of D5S2500 were observed in the samples respectively, the observed heterozygosity (Ho) values, the discrimination power (DP) values and the power of exclusion (PE) values of D2S1399 and D5S2500 is 0.745 and 0.807, 0.958 and 0.917, 0.554 and 0.643, respectively. CONCLUSION The result showed that D2S1399 and D5S2500 were highly informative loci and suitable for forensic application.
Alleles ; Asian People/genetics* ; Electrophoresis, Polyacrylamide Gel ; Forensic Medicine ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Silver Staining ; Tandem Repeat Sequences

AIM To observe the effects of Sangtongjian Mixture (STJ) on glucose and lipid metabolism,insulin resistance and fat cytokines in type 2 diabetic rats,and their mechanisms of action.METHODS One hundred and forty rats fed on the combination of STZ and high fat diet were established as the type 2 diabetic models.Fasting blood glucose (FBG) level reached more than 16.7 mmol/L and then the rats were randomly divided into model group,metformin (180 mg/kg) group,STJ (73.5,147 and 294 mg/kg) groups.Ten rats were set as the blank group.Each treatment group was intragastrically given the corresponding agents for twelve weeks.The fasting blood glucose levels of rats were measured once every two weeks after the administration.After a 12-week administration period,glycosylated serum protein (GSP),glycosylated hemoglobin (GHb) and lipid profile indices (TC,TG,HDL-C and LDL-C) were determined.The serum insulin level was measured by radioimmunoassay,and homeostasis model assessment of insulin resistance (HOMA-IR) and insulin sensitivity index (ISI) were calculated.The levels of serum adiponectin and leptin were detected by ELISA.RESULTS STJ remarkably decreased the levels of FBG,GSP,GHb,TC,TG,LDL-C,leptin and HOMR-IR in type 2 diabetic rats.Furthermore,STJ also significantly increased the levels of HDL-C,adiponectin and ISI.CONCLUSION STJ can improve glucose and lipid metabolism in type 2 diabetic rats by ameliorating insulin resistance and regulating fat cytokine levels.

OBJECTIVETo investigate the effects of interferon-gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-gamma) on the sperm acrosin activity and the rate of acrosome reaction and to probe into their mechanisms.

METHODSThirty-six nearly normal semen samples were treated with IFN-gamma and/or TNF-alpha after isolated by 75% Percoll. The sperm acrosin activity was tested by the method of BAEE/ADH Unity, the rate of acrosome reaction observed by Triple-stain technique, the NO concentration measured by HPLC and the activities of Na+ -K+ -ATPase, Ca2+ -ATPase and SOD assayed by kit method.

RESULTSBoth IFN-gamma and TNF-gamma could decrease sperm acrosin activity and acrosome reaction (P < 0.05 or P < 0.01). TNF-alpha showed stronger inhibiting effect, IFN-gamma markedly reduced the activities of Na+ -K+ -ATPase, Ca2+ -ATPase and SOD in sperm (P < 0.01), and their synergistic action was weaker. However TNF-alpha produced hardly any effect on Na+ -K+ -ATPase and Ca2+ -ATPase. The NO concentration in sperm was significantly increased by IFN-gamma and/or TNF-alpha (P < 0.01).

CONCLUSIONIFN-gamma and TNF-alpha have some inhibiting effect on sperm acrosin activity and the rate of acrosome reaction, which could be attributed to their influence on the activities of Na+ -K+ -ATPase, Ca2+ -ATPase and SOD, the NO concentration and so on.

Acrosome Reaction ; drug effects ; Adult ; Calcium-Transporting ATPases ; metabolism ; Chromatography, High Pressure Liquid ; Humans ; Interferon-gamma ; pharmacology ; Male ; Middle Aged ; Nitric Oxide ; metabolism ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Spermatozoa ; drug effects ; enzymology ; metabolism ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo establish a green-fluorescent protein (GFP) labeled tumor metastasis model and to evaluate its biological characteristics.

METHODSHuman gastric carcinoma cell MGC-803 and murine cervical carcinoma cell U14 were transfected with the plasmid pEGFP-N1 and the efficiency of transfection was assessed 24 h later. Limited dilution was employed to screen and establish monoclonal cell strains, MGC-803-GFP and U14-GFP. The two fluorescent tumor cell stains were transplanted into BALB/c-nu mice and C57BL/6J mice respectively. The latency period of tumor mass appearance and the growth curve in vivo were documented. The tumor growth and metastasis were evaluated in vivo by the Viviperception Fluorescence Imagining System (VFIS). Expressions of CD44 and E-cadherin in tumor tissue were monitored by immunohistochemistry.

RESULTSThe efficiency of pEGFP-N1 transfection of MGC-803 cells and U14 cells were 30% and 60%, respectively. Monoclonal GFP(+) cell strains-MGC-803-GFP and U14-GFP were established. The latency periods of tumor formation of MGC-803-GFP and U14-GFP were 3-5 days and 2-4 days, respectively. Their tumorigenicity rates were 100% in both. The tumor growth of MGC-803-GFP was well defined by the VFIS. Only one mouse was shown to harbor lymphatic metastasis by VFIS, 60 days after transplantation. The metastasis process of U14-GFP was depicted through VFIS on 27, 37 and 52 days post-transplantation. The incidence of pulmonary metastasis and lymphatic metastasis of U14-GFP was 67% and 100% respectively when the tumor volume was >or=5 cm3. CD44 was positive and E-cadherin was negative in both tumors by immunohistochemistry.

CONCLUSIONSSuccessfully established two monoclonal tumor cell strains stably expressing GFP: MGC-803-GFP and U14-GFP. Transplantation of these cells into mice can establish tumor metastasis models which could be used for future visualized tumor research in vivo.

Animals ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; secondary ; Lymphatic Metastasis ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Nude ; Neoplasm Transplantation ; Plasmids ; Stomach Neoplasms ; metabolism ; pathology ; Transfection ; Tumor Burden ; Uterine Cervical Neoplasms ; metabolism ; pathology

OBJECTIVETo elucidate the expression and function of VAP-33 gene in dendritic cell sarcoma (DCS) cell line.

METHODSThe expression of VAP-33 in DCS cells was investigated by mass spectrum with immunoprecipitation membrane protein. DCS cells were treated with antigens in different dosages (150, 850, and 1500 microl) for 24, 48 and 72 h respectively. Cell morphology and phagocytosis activity of DCS cells were measured. Indirect immunofluorescence, confocal microscopy and Western blotting were used to study the distribution and expression changes of VAP-33. Moreover, DCS cells were treated with 0.5 mol/L insulin for 20 min first and followed by Western blotting to detect changes of VAP-33 and glucose transfer protein 4 (GLUT-4) in the total cellular protein, cytoplasmic protein and membrane protein. Confocal microscopy was used to document the expression and distribution changes of VAP-33 and GLUT-4 in DCS cells.

RESULTSVAP-33 expression was obtained at the cell membrane and in the cytoplasm of DCS cells. Upon antigen stimulation, DCS cells showed more active phagocytosis and morphologically became more elongated with branched protrusions. The expression of VAP-33 was decreased by the antigen stimulation. Upon the insulin stimulation, the expression of VAP-33 and GLUT-4 were increased and co-localized.

CONCLUSIONSVAP-33 expression in DCS originated from the dendritic cells (DC) seemed relating to the vesicle transportation during antigen processing in DC. Additionally, VAP-33 and GLUT-4 also take part in the glucose transportation in the cells.

Animals ; Antigen Presentation ; Carrier Proteins ; metabolism ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Cytoplasm ; metabolism ; Dendritic Cell Sarcoma, Interdigitating ; metabolism ; pathology ; Down-Regulation ; Glucose Transporter Type 4 ; metabolism ; Insulin ; pharmacology ; Membrane Proteins ; metabolism ; Mice ; Phagocytosis ; immunology

OBJECTIVETo investigate the role that E-cadherin (E-cad) plays on cell adhesion and proliferation of human breast carcinoma.

METHODSE-cad expression vector was transfected into an E-cad-negative human breast carcinoma MDA-MB-231 cells. G418 was used to screen positive clones. E-cad, β-catenin (β-cat) and cyclin D1 expressions of these clones were confirmed by Western blot. Their cell-cell and cell-matrix adhesion abilities were detected. E-cad/β-catenin interaction was confirmed by immunoprecipitation. Cell proliferation was evaluated by MTT. Cell apoptosis was analyzed by flow cytometry. Direct two-step immunocytochemistry was used to detect the localization of β-cat.

RESULTE-cad(+) cell strains Ecad-231-7 and Ecad-231-9 were established. When cultured in ultra-low-binding dishes Ecad-231 cells grow in suspension while Ecad-231-7 and Ecad-231-9 cells grow in large clamps. When co-cultured with HCT116 cells, the average adhesion rates at 30 min are 39.0%, 60.0% and 59.5% for MDA-MB-231, Ecad-231-7 and Ecad-231-9 respectively. The average detachment rates by EDTA for 5 min are 37.4%, 4.2% and 7.4% respectively. So E-cad expression enhanced hemotypic and heterotypic cell-cell adhesion and cell-matrix adhesion. Forced exogenously expressed E-cad could combine with endogenous β-cat, whereas down stream cyclin D1 expression was significantly decreased, as evidenced by Western blot. The rates of cell apoptosis of MDA-MB-231, Ecad-231-7 and Ecad-231-9 were 1.8%, 2.0% and 2.1%. Expression of E-cad had no obvious effect on the apoptosis of tumor cells with regular culture. β-cat increased in the cytoplasma.

CONCLUSIONSTwo monoclonal tumor cell strains (Ecad-231-7 and Ecad-231-9) stably expressing E-cad were successfully established. E-cad could enhance adhesion and inhibit proliferation of human breast carcinoma cells through a pathway involving β-cat and cyclin D1.

Apoptosis ; Breast Neoplasms ; metabolism ; pathology ; Cadherins ; genetics ; metabolism ; physiology ; Cell Adhesion ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Female ; Genetic Vectors ; Humans ; Plasmids ; Transfection ; beta Catenin ; metabolism