Child ; Child Development ; drug effects ; Endocrine Glands ; drug effects ; growth & development ; physiology ; Hazardous Substances ; poisoning ; Humans ; Nervous System ; drug effects ; Urogenital System ; drug effects

AIM　To study whether KB-R7943 has selective inhibitory effect on the inward and outward Na+-Ca2+ exchange current (INa-Ca) in guinea pig ventricular myocytes. METHODS　Through setting up the model of intracellular Na+-overload during myocardial ischemia and reperfusion, the current-voltage relationship of INa-Ca was recorded using whole-cell patch clamp technique under bi-directional ionic conditions. RESULTS Currents were elicited by a declining ramp pulse depolarized immediately from holding potential of -40 mV to +60 mV, then repolarized to -100 mV at a speed of 80 mV*s-1 and returned to the holding potential under bi-directional ionic conditions, while the ［Na+］ was 25 mmol*L-1 in the pipette solution. The currents increased time-dependently and voltage-dependently which reached from (2.51±0.15) pA*pF-1 to (5.94±0.13) pA*pF-1 at +50 mV and from (-1.92±0.13) pA*pF-1 to (-3.17±0.16) pA*pF-1 at -80 mV (n=12) after 3 min and there is no significant run-down of the current. KB-R7943 10-6 mol*L-1 was found to decrease the current to (4.62±0.05) pA*pF-1 by 29.4% at +50 mV and to (-2.30±0.18) pA*pF-1 by 22.1% at -80 mV (n=5) after 5 min. While 10-5 mol*L-1 KB-R7943 was shown to decrease the current to (3.13±0.03) pA*pF-1 by 61.7% at +50 mV and to (-1.62±0.03) pA*pF-1 by 56.9% at -80 mV (n=7). CONCLUSION　KB-R7943 can block INa-Ca in guinea pig ventricular myocytes. But, it did not show selective inhibition effect on inward and outward currents.

Adult ; Cord Blood Stem Cell Transplantation ; adverse effects ; Cytomegalovirus ; Cytomegalovirus Infections ; complications ; Female ; Hemolysis ; Humans

AIM: To establish the determination of ginsenoside R e and R g1 in Compound Jiangtang Oral Liquid(Radix Ginseng, Fructus Schisandrae Chinensis, Radix Rehmannia, Radix Asparagi, etc.) by HPLC. METHODS: C 18 ODS was used as a stationary phase, acetonitrile-0.05% phosphoric acid (96∶400) as a mobile phase and detection wavelength at 203nm. RESULTS: The linear range of ginsenoside R e concentration was from 0.66 to 3.29?g?mL -1 and correlation coefficient was 0.9991, the linear range of ginsenoside R g1 concentration was from 1.05 to 5.27?g?mL -1 and correlation coefficient was 0.9998. The average recovery of sample was 98.2% and RSD 2.2% ( n =5), respectively. CONCLUSION: The method is simple and convenient, accurate with a good reproducibility and can be used for determination of ginsenoside R e and R g1 in Compound Jiangtang Oral Liquid.

Objective To observe how the ACC transmits nociceptive information and how it regulates spinal noceciption.Methods A total of 32 male SD rats were randomly divided into four groups:Sham group,CCI group,ACC group,and AP-5 group.After light-dark transition test, forced swimming test (FST),paw withdrawal mechanical threshold (PWMT),paw withdrawal ther-mal latency (PWTL)had been measured,the rats were finally anesthetized,then ACC and the spinal cord was rapidly removed,the expression of cAMP-response element binding protein (pCREB)and extracellular regulated protein kinases (pERK)were measured by Western blot.Results Compared with the sham group,the PWMT of the CCI rats were significantly decreased,rats spent less time in the light compartment and number of transition were decreased (P <0.01 ).The immobility time in FST were also significantly prolonged (P <0.01).After AP-5 injected in bilateral ACC 13 days after CCI operation,the PWMT of the CCI rats were significantly increased,rats spent more time in the light compartment and number of transition were increased (P <0.01).The immobility time in FST were also significantly shortened (P <0.01).Compared with sham group,the expression of pCREB, pERK increased significantly in ACC in CCI group (P <0.01).The expression of pCREB,pERK in spinal cord was increased in CCI group and decreased in AP-5 group (P <0.01 ).Conclusion ACC facilitate the spinal nociception in a descending mode.

Objective To assess alternations in left ventricular volume and systolic synchrony in patients with frequent premature ventricular complexes(PVCs) from the right ventricular outflow tract(RVOT).Methods Twenty-nine patients with frequent isolated PVCs from RVOT were included and 30 healthy subjects as control.Instantaneous full-volume imaging(IFI) was performed to evaluate left ventricle volumetric parameters,including end-systolic volume (ESV),end-diastolic volume (EDV),stroke volume (SV),ejection fraction (EF),and systolic synchrony parameters,including systolic dyssynchrony index (SDI),dispersion end-systole (DISPES),mean end-systolic time (MES),pre-contraction time volume (PreContr) and post-contraction time volume (PostContr).Contraction front mapping was performed to visualize volumetric contraction sequence.All values of patients with PVCs were recorded during sinus beats (PVC-S) and premature ventricular beats (PVC-V) respectively.Results Significant differences were observed in left ventricular systolic volumetric and synchrony parameters between PVC-V and control subjects (P＜0.01),as well as in MES and PreContr between PVC-S and control subjects (P＜0.01).Conclusions Left ventricular systolic dysynchrony was demonstrated in patients with PVCs from RVOT.IFI was a novel tool to analyze left ventricular global and regional volumetric alternations.

Chinese cobra (Naja naja atra) cardiotoxins are three-fingered family with 60～62 amino acids bind by four disulfide bonds. CardiotoxinⅢ (CTXⅢ) is one of the major toxic component which can cause hemolysis and cytotoxicity. However, there is no report on the fusion expression of CTXⅢ in soluble form so far. The cloning, expression and purification of recombinant CTX Ⅲ (rCTXⅢ) from Naja naja atra in E. coli and in yeast Pichia pastoris were reported here. CTXⅢ gene, fused with enterokinase in E.coli His-patch Thioredoxin expression system, were expressed in soluble form and released by osmotic-shock treatment. CTX Ⅲ gene was also cloned and expressed in the methylotropic yeast Pichia pastoris pPIC9K expression vector in the first time. The yield of the secretion level was 9.5 mg/L. Using straightforward one-step chromatography procedure, the rCTXⅢ, with three additional amino acids (GYT) at the N-terminal site, was purified to a purity of more than 90% and recovery yield of 65%. The purified rCTX Ⅲ was further characterized by cytotoxic assay with IC50 4.66μg/ml. An effective expression and purification system for recombinant CTXs in P. pastoris was developed, this system will permit us the ready isolation of active cardiotoxins. This protocol can also be easily used for the production of the toxin in a larger scale with low cost.

Objective To establish the sperm specific Sleeping Beauty ( SB ) transposase-expression transgenic mouse for the study of the genetic modification mediated by transposon system in mouse .Methods Prm1 promoter was cloned from mouse genomic DNA to drive the expression of SB transposase .The transgenic mice were generated by microinjection .The gene type of transgenic line was identified by PCR .The expressing level in testis was determined by western blot and immunohistochemistry (IHC) staining.Results Five lines of transposase transgenic mice were obtained by microinjection and three can be germline .One mouse line with higher expression level of transposase in the testis was obtained.Conclusion One transgenic mouse model with Sleeping Beauty transposase - expression was successfully established .This model will greatly contribute to the research of genetic modification mediated by transposon in mouse.

Objective To observe the acute skin and mucous membrane reactions in patients treated with concurrent radiochemotherapy for nasopharyngeal carcinoma,and to analyze the influencing factors.Methods A total of 85 nasopharyngeal carcinoma cases treated with concurrent radiochemotherapy were enrolled in the study.Fifteen clinical and laboratory indexes,including BMI,radiation dose,degree of acute oral mucous and skin reactions and blood routine test were observed weekly.Univariate and multivariate regression analysis were performed to assess the factors,and screen the independent factors.Results Multiple-factor analysis showed that the risk factors cloesly related with acute radioactive oral mucosa reactions were smoking history(OR =3.467,P ＜ 0.05),single-dose of gross tumor volume (GTV) ＞2.15 Gy(OR =3.393,P ＜ 0.05),while those with acute radiation skin reactions were diabetes history(OR =87.859,P ＜ 0.05) and hemoglobin values 1 week before radiotherapy ＞ 130 g/L (OR =21.404,P ＜ 0.05).Conclusions In the patients treated with concurrent radiochemotherapy for nasopharyngeal carcinoma,smoking history and single-dose of GTVnx is the independent risk factors of acute radiation oral mucosa reactions,while diabetes history and hemoglobin values I week before radiotherapy are the independent factors of acute skin reactions.

Objective To investigate the effects of β-arrestin1 on proliferation,migration,invasion and apoptosis of human gastric cancer BGC-823 cell line.Methods The expression of β-arrestin1 in human gastric epithelial cell line GES,human gastric cancer cell line BGC-823,MKN-28 and SGC-7901 was detected by realtime-polymerase chain reaction (PCR) and Western blot.The stable β- arrestin1 and negative control interfered BGC-823 cell line were established by RNA interference technology.The cell proliferation,migration,invasion and cell apoptosis of β-arrestin1 stable interfered BGC-823 cell line was examined by cell counting,scratch test,Transwell chamber test and flow cytometry assays.The data were analyzed by t test.Results The expression of β-arrestin1 in cell line GES,MKN-28,SGC-7901 and BGC-823 was 0.001 ± 0.001,0.002 ± 0.000,0.003± 0.002 and 0.005 ± 0.000 respectively.The inhibition ratio of proliferation in β-arrestin1 interfered BGC-823 cells and negative control cells were -30.2 ％ and 100.0 ％.The invasion ability was also inhibited,the number of migratory cells was 126.25±3.24 and 213.50±6.27 (t=0.000,P＜0.01),and the apoptosis rate was (41.350±1.053)％ and (11.497±0.589) ％ (t=0.015,P＜0.05).Conclusions β-arrestin1 is highly expressed in gastric carcinoma,and the expression increased along with the malignancy degree.The cell proliferation,migration and invasion is inhibited by interference of β-arrestin1 in BGC-823 cells,while the cell apoptosis is promoted.