ObjectiveTo explore the influence of electrical stimulation on nervous cells.MethodsOn 5th day the brain primitive nervous cells of rat brain, in vitro culturing, were divided into 4 groups, in order of control group, electrical pole control group, low stimulation group, and high stimulation group. The cells were stimulated with electro-acupuncture apparatus for 7 days, 30 minutes once, twice a day. On 10th day, the cells were observed microscopically for 3 days and taken photos.ResultsElectrical stimulation is helpful to raise the survival rate of cell.Conclusions Electrical stimulation influences morphological change of nervous cells during culturing.

Objective To investigate the association of ret gene rearrangement mutation with the pathogenesis of papillary thyroid carcinoma (PTC). Methods Twenty-seven cases of PTC were analysed for expression of ret gene rearrangement (ret/PTCs) by multiplex-PCR at first, and then ret/PTC1-3 were identified by identification-PCR (ID-PCR). Finally, the specific ret/PTC was affirmed by automated direct sequencing. Ten specimens of malignant thyroid tissues of other histological types, 33 benign thyroid lesions and 30 normal thyroid specimens beside tumor (as the control) were also included. Results (1) Fifteen samples showed positive ret/PTCs, 11 of which harboured ret/PTC1,3 were positive for ret/PTC3, and 1 for ret/PTC2. All the rearrangements were clearly identified by automated direct sequencing of ID-PCR products. (2) All the 15 ret/PTC-positive tissue samples were histologically confirmed to be PTC. The prevalence of the tumor-specific ret rearrangements in 27 patients with PTC is 55.6% (15/27). (3) No significant difference was found regarding the gender and age of the patients, tumor size and metastasis of neck lymph node. 33.3% (2/6) of the PTC with invasion of extrathyroidal soft tissue were ret/PTC-positive, as compared with 66.7%(4/6) for ret/PTC-negative group (P

Actins ; immunology ; metabolism ; Antibodies, Monoclonal ; metabolism ; Child ; Diagnosis, Differential ; Fibroma ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Gastrointestinal Neoplasms ; metabolism ; pathology ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; Humans ; Leiomyoma ; metabolism ; pathology ; Male ; Pyloric Antrum ; pathology ; Stomach Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

This study was performed to investigate the chemical constituents in the twigs and leaves of Harrisonia perforate. Six compounds were isolated from the 95% EtOH extract of the twigs and leaves of Harrisonia perforate by silica gel, ODS, Sephadex LH-20 column chromatographies and preparative HPLC. On the basis of chemical properties and spectra data, these compounds were identified as harriperfin E (1), kihadanin A (2), kihadanin B (3), 6α-acetoxyobacunol acetate (4), gardaubryone C (5), and β-sitosterol methyl ether (6), respectively. Compound 1 is a new chromone, and compounds 2-6 are isolated from this plant for the first time.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Phytochemicals ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Simaroubaceae ; chemistry

Adolescent ; Child ; Child, Preschool ; Cytomegalovirus Infections ; complications ; Female ; Humans ; Kidney Diseases ; etiology ; Kidney Glomerulus ; pathology ; Male

Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; metabolism ; Protein Isoforms ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; Tumor Cells, Cultured

Objective To investigate the apoptosis of T lymphocytes,the expression of Bcl-2, Fas on the peripheral blood T lymphocytes in end stage renal disease patients;and to explore the characteristics of Th1 /Th2 profile and the influence of dialysis membranes with different permeability on the apoptosis of T lymphocytes of maintenance hemodialysis patients.Methods The study included 10 non-dialyszed (ND)patients,45 maintenance hemodialysis patients with cellulose acetate (CA) membranes(13),low-flux polusulfone(PS-LF) membranes(16),high-flux polusulfone (PS-HF) membranes (16) and 8 healthy volunteers (C).The apoptosis of T lymphocytes,expression of Bcl-2,Fas on peripheral blood T lymphocytes cultured with phytohemagglutinin (PHA) stimulation for 24 hours were measured by flow cytometry and immunohistochemical.ELISA was performed for detecting the levels of IFN-?and IL-4 in culture supematants.Results In ESRD patients,the apoptosis of T lymphocytes was greater than that of group C.Group CA was greater than group PS-HF and group PS-LF (P＜0.05).The expression of Bcl-2 on T lymphocytes in ESRD patients was lower than that of group C (P＜0.05).There was negative correlation between the T lymphocytes apoptosis and Bcl-2. The expression of Fas on T lymphocytes in ESRD patients was greater than that of group C (P＜0.05), and it was positive correlated with T lymphocytes apoptosis.The level of IFN-?of ESRD patients was decreased significantly compared with that in group C (P＜0.05),and there was negative correlation between T lymphocytes apoptosis and IFN-?.IL-4 was increased in ESRD patients (P＜0.05) and it was positive correlated with T lymphocytes apoptosis.Conclusions The accelerated apoptosis of T lymphocytes in ESRD patients may be related to the expression of Bcl-2 and Fas of T lymphocytes.ESRD patients show a suppressed secretion of IFN-?and an increased secretion of IL-4. T lymphocytes apoptosis of maintenance hemodialysis patients is influenced not only by the biocompatibility but also by the permeability of the dialysis membrane.

Objective To establish real-time PCR with SYBR Green Ⅰ for quantification the gene of survivin.Methods The components and conditions of PCR system were optimally determined by fluorescence intensity,cycle threshold(Ct),melting curve,coefficiency and slope of the standard curve.The means to eliminate the contaminating fluorescence of primer-dimers and the mode for Ct value determination were also optimized.Using the developed PCR system,we quantificated the survivin gene in 43 patients with gastric carcinoma.Results The optimized condition for PCR amplification of survivin were 2 mmol/L of MgCl_2,2.5 U/100 ?l of Taq DNA polymerase,0.2 ?mol/L of primers,and the optimized annealing temperatures for PCR were 58℃.The influence of primer dimmer can be eliminated by setting the fluorescence collecting temperature below the Tm of the specific amplicon by 2℃.The second derivative maximum mode,instead of fit point mode,was a feasible method to determine the Ct value for quantification.The sensitivity of this method was 10 copies/?l,and a good linearity was found from 10~1 to 10~4 copies/?l(r = 0.999 7).The inter-experimental coefficient of variation was 1.13%-1.91%,whereas the coefficient of variation between runs was 3.31%-4.50%.Using the optimized PCR system,we quantificated the gene of survivin,the result indicated that survivin gene was amplified in 13.9% of gastric carcinomas.Conclusions The optimal real-time PCR with SYBR Green Ⅰ,as a cost-effective and feasible DNA quantitative method,is fit for quantification of the survivin with satisfactory repeatability and high sensitivity.

Objective To investigate the relationship between refractory hypertension and renal hemodynamics in end stage renal diseases (ESRD) patients.Methods ESRD patients were classified into:patients with refractory hypertension (group A) and patients with normal blood pressure(group B).Renal hemodynamic indices were ex- amined by duplex ultrasonography.Fasting serum lipid (TC,TG,HDL-C,LDL-C,Lp(a),ox-LDL) and serum parathyroid hormane (PTH) were determined in all patients.Results Significant differences were found in renal hemodynamic indices such as peak systolic velocity (PSV),mean flow velocity (MV),pulsatility index (PI),renal- aortic ratio (RAR) and in clinical index such as Lp(a) and ox-LDL between the two group.Refractory hyperten- sion patients had lower renal hemodynamic indices and higher Lp(a) and ox-LDL levels than in patients with con- trolled BP.Logistic regression analysis revealed that refractory hypertension was related with PSV,EDV,Pl and RAR,but not relevant with sex,age,dialysis time,hematocrit,BUN,creatinine,TC,TG,HDL-C,LDL-C, PTH,MV and RI.Conclusion Atherosclerotic renal artery stenosis and severe disorder in renal hemodynamics is likely the cause for refractory hypertention in ESRD patients.The rise of serum Lp(a) and ox-LDL might acceler- ate renal artery atherosclerosis.

Objective To construct an eukaryotic expression vector with human adipose most abundant gene transcript 1 (APM1) gene,and to investigate the transfection and expression of pCDEF-APM1 eukaryotic expression plasmid in HEK293 cells.Methods pCDEF-APM1 eukaryotic expression plasmid was constructed by DNA recombinant method.Expression vector pCDEF-APM1 was transfected into HEK293 cells with Effectene reagent.The level of human adiponectin protein in the supernatant of cell culture media was detected with double antibody sandwich ELISA.Results The sequence of DNA fragment from constructed pCDEF-APM1 plasmid was identical to that published in GenBank.There was raised human adiponectin protein level in culture supernatant of HEK293 cells tnmsfected with pCDEF-APM1.Conclusion The pCDEF-APM1,an eukaryotic expression plasmid for APM1 gene is successfully constructed.High protein expression of adiponectin can be obtained in HEK293 cells transfected with pCDEF-APM1 eukaryotic expression plasmid.