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

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 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 expression features of glypican-3 (GPC-3) and its diagnostic and differential values in hepatocellular carcinoma (HCC).

METHODSRat hepatoma models were made and the dynamic expression features of GPC-3 protein and its gene were investigated by Western blotting and RT-PCR respectively. Liver specimens from 36 HCC patients were collected by self-control method and the expression and clinicopathological features of GPC-3 were analyzed by immunohistochemistry. Serum GPC-3 levels were quantitatively detected by ELISA and its efficiency for HCC diagnosis was evaluated in patients with liver diseases.

RESULTSThe incidence of GPC-3 was 0% in control, 83.3% in degeneration, 100% in precancerosis and 100% in canceration during dynamic formation of rat hepatoma, respectively. The positive GPC-3 was brown granule- like staining localized in membrane and cytoplasm in human HCC.

CONCLUSIONSThe GPC-3 positive rates were 80.6% in HCC, 41.7% in surrounding tissues and none in distal tissues (P < 0.01), respectively. No positive relationship presented between GPC-3 and differentiation grade or the number of tumor except of tumor size (Z = 2.941, P < 0.01). The incidence of serum GPC-3 was 52.8% in HCC patients except of one patient with cirrhosis. No significant differences were found between GPC-3 and sex, age, AFP, tumor number, Child classification or extrahepatic metastasis except of tumor size (χ² = 6.318, P < 0.05) and HBV infection (χ² = 23.362, P < 0.01). Combined detection of GPC-3 and AFP could rise up diagnosis of HCC. GPC-3 expression closely associated with HCC and might be useful for early diagnosis of HCC.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Carcinoma, Hepatocellular ; diagnosis ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Glypicans ; metabolism ; Humans ; Liver ; pathology ; Liver Neoplasms ; diagnosis ; metabolism ; pathology ; Male ; Middle Aged ; Rats ; Rats, Sprague-Dawley ; Young Adult

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

OBJECTIVETo explore the feasibility of using enriched bone marrow (BM) compound with fibrin glue (FG) in repairing old radial bone defect.

METHODSTotally 36 New Zealand rabbits were equally randomized into three groups: simple FG group, BM+FG group, and enriched BM+FG group. A 1.5-cm segmental bone defect was made at the left radial in each animal. After one month, the defect was implanted with the engineered bone. Before implantation, a compound of enriched BM with FG underwent electron microscopy, long-term culture, and bacteriological culture. Four, 8, and 12 weeks after operations, the osteogenetic effect was evaluated using X-ray observation, HE staining, or Van Gieson staining, and a semi-quantitative analysis was performed.

RESULTSElectron microscopy showed enriched BM were compatible well with FG. No bacterial contamination or oncogenicity was observed after long-term culture. X-ray showed the repair effectiveness was significantly higher in BM+FG group and enriched BM+FG group than in simple FG group. Eight and 12 weeks after surgery, the Yang scores were significantly higher in enriched BM+FG group than in BM+FG group [(9.348±0.364évs.(7.984±0.229éìF=40.167ìP=0.001; (12.664±0.388)vs. (10.584±0.836é, F=20.3647ìP=0.004]. In addition, the Yang's scores at bone defects in BM+FG group and enriched BM+FG group were higher at the 12(th) week than in the 8(th) week. (F=36.004ìP=0.001; F=155.141ìP=0.000; respectively)The bone defects were repaired at varied degrees were histologically observed in BM+FG group and enriched BM+FG group during the observations.

CONCLUSIONImplantation of BM+FG or enriched BM+FG are effective in repairing old radial bone defects, while simple FG shows not such effect.

Animals ; Bone Marrow ; Bone Substitutes ; Disease Models, Animal ; Female ; Fibrin Tissue Adhesive ; Male ; Rabbits ; Radius ; injuries ; surgery ; Tissue Engineering