Objective To establish the quality standard for Haijinhuwei powder. Methods Corydalis Rhizoma, Angelicae Dahuricae Radix, Angelicae Sinensis Radix were identified by TLC. The content of tetrahydropalmatine was determined by HPLC. Results TLC spots were clear and specific.There was a good linear relationship between peak area and concentration of tetrahydropalmatine at the range of 10.78-107.8μg.mL-1(r=0.999 9).The average recovery rate was 96.57% and RSD was 1.40%(n=6). Conclusion All these results indicated that the developed TLC-HPLC method was proved to be reliable, accurate and specific, which could be used for the quality control of Haijinhuwei powder.

Objective To construct a recombinant lentiviral expression vector containing NIS and EGFP gene,and to explore the feasibility of NIS gene for monitoring the bone marrow derived mesenchymal stem cells (BMSCs) migration to the intracranial glioma.Methods The NIS and EGFP gene fragments were subcloned into lentiviral vector pLVX-puro,then packaged and amplified in HEK293T cells to obtain recombinant lentivirus pLVX-CMV-NIS-EGFP.pLVX-CMV-0-EGFP was constructed as control.BMSCs were isolated,cultured,and transfected by lentivirus.The antibiotic-resistant transfected BMSCs (BMSCs-NIS-EGFP and BMSCs-EGFP) were selected.The expression of NIS gene was examined by Western blot.Functional NIS activity was confirmed by the uptake of 125I and the inhibition effect of NaClO4.The nude mice intracranial glioma models were established.MicroSPECT was performed at 24 h post BMSCs-NIS-EGFP injection via the tail vein.Results pLVX-CMV-NIS-EGFP and pLVX-CMV-0-EGFP were successfully constructed and packaged.BMSCs were successfully isolated and cultured.Stable cell lines BMSCs-NIS-EGFP and BMSCs-EGFP were constructed after lentivirus transfection and puromycin selection.The expression of NIS gene was detected by Western blot in BMSCs-NIS-EGFP,but not in BMSCs-EGFP.BMSCs-NIS-EGFP showed significantly more uptake of 125I (nearly 10 times than the uptake in BMSCs-EGFP) and the uptake could be significantly inhibited by NaClO4.The nude mice intracranial glioma models were successfully established and the BMSCs-NIS-EGFP in glioma foci could be visualized by microSPECT imaging at 24 h post injection.Conclusions A recombinant lentivirus containing NIS gene could be successfully constructed for monitoring BMSCs migration towards intracranial glioma.It might provide evidence on the research of BMSCs and NIS gene mediated therapy for glioma.

Objective:To study the expression of caspase-3 and free radical injury in hypoxic-ischemic brain damage ( HIBD) of neonatal rats.Methods:All seven-day-old Wistar rats were randomly divided into HIBD group and sham operation group.Brains was obtained at time of 6 h,12 h,24 h,48 h,72 h,96 h after HIBD.Neuronal cell apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling ( TUNEL).The expression level of caspase-3 protein was detected by immunohisto-chemistry.The level of malondialdehyde ( MDA) and supemxide dismutase ( SOD) were measured by thiobarbitic acid colorimetry and xanthin oxidase,respectively.Results:The number of neuronal cell apoptosis and the expression of caspase-3 protein began to increase at 6 h and reached the peak at 48 h in HIBD group,they were both significantly higher than those in the sham operation group at each time point (P<0.05).The level of MDA began to increase at 6 h and reached the peak at 24 h in HIBD group,it was significantly higher than the sham operation group at each time point (P<0.05).The level of SOD began to decrease at 6 h and reached the lowest level at 24 h in HIBD group,it was significantly lower than the sham operation group at each time point ( P<0.05 ) .The number of neuronal cell apoptosis and the expression of caspase-3 protein were positively correlated with the level of MDA, but they were negatively correlated with the level of SOD.Conclusion: Free radical injury promotes the expression of caspase-3 and neuronal cell apoptosis in HIBD.

Objective To observe the effect of scalp acupuncture and rehabilitation training on the regulation of neuro-plasticity-associated protein MAP-2 in gerbils modelling cerebral ischemia and reperfusion.Methods Cerebral ischemia and reperfusion were modelled into thirty-six male gerbils.They were then randomly divided into a model group (A),a scalp acupuncture and rehabilitation training group (B) and a rehabilitation training group (C),each of 12 cases.The scalp acupuncture and rehabilitation training were administered daily to the gerbils in group B beginning 24 h after modeling,for 14 d.Any changes in Bederson scores were observed after one,7 and 14 days.On the 14th day the expression of MAP-2 around the infarct focus was detected using western blotting in all 3 groups.Results On the 7th day the average Bederson score in group B (1.81 ± 0.52) was not significantly different from that in group A (2.13 ± 0.49) or group C (2.00 ± 0.31) (P ＞ 0.05).At the end of the treatment,however,there were significant differences between the groups,with group B (0.47 ± 0.31) scoring significantly better than groups C (1.04 ± 0.63) and A (1.46 ± 0.72) (P ＜ 0.05).Group C was also significantly better than group A (P ＜0.05).The expression of MAP-2 as measured as integrated optical intensity (IOD) in group B (0.91 ±0.18) was significantly higher than that in group A (0.43 ± 0.21) and group C (0.67 ± 0.24) (P ＜ 0.05) ; so was group C compared to group A (P ＜0.05).Conclusion The scalp acupuncture combined with rehabilitation training can improve the recovery of motor function and enhance the expression of neuro-plasticity-associated protein MAP-2 in gerbils with after cerebral ischemia and reperfusion.

85 mL/min),CL=6.0?(WT/60)~(0.52).④The increased volume of peripheral distribution (V_2) was observed when norvancomycin was co-administered with diuretics;④Reduced drug clearance,prolonged t_(1/2),and increased values of AUC_(24) were found in elderly patients.Conclusions Renal function impairment and age have significant impact on PK parameters of norvancomycin.Dosing regimens of norvancomycin were finally established for different patients on the basis of important PPK parameters generated from different groups of patients.

OBJECTIVETo determine the prevalence of saliva Helicobacter pylori in Lanzhou and investigate Helicobacter pylori-related diseases.

METHODSHelicobacter pylori was detected through bacterial culture, Gram stain microscopy, and urease test from saliva samples collected from 941 residents of Lanzhou. The infection rate and growth of Helicobacter pylori among the residents were analyzed in terms of different oral health conditions, oral disease, gender, urban and rural status, and age.

RESULTSThe rate of Helicobacter pylori-positive saliva in Lanzhou was 42.72%. The status of Helicobacter pylori infection showed significant difference among subjects with different oral hygiene and oral diseases. The rate of Helicobacter pylori-positive saliva among females was 47.89%, which was greater compared with the rate among males (38.45%, P = 0.004, chi2 = 8.492). The rate of Helicobacter pylori-positive saliva in the town was 33.99%, which was less than the rate for the villages (50.93%, P = 0.000, chi2 = 27.551). The rate of Helicobacter pylori-positive saliva among residents aged 10 to 59 showed a flat trend with no significant differences. However, the rate of Helicobacter pylori-positive saliva among residents over 60 years old showed a significant increase. No significant difference was found in the growth of saliva Helicobacter pylori (P = 0.086).

CONCLUSIONThe rate of Helicobacter pylori-positive saliva is related to the subjects' oral hygiene, oral disease, gender, age, and living conditions.

Adolescent ; Adult ; Child ; China ; epidemiology ; Female ; Helicobacter Infections ; epidemiology ; Helicobacter pylori ; Humans ; Male ; Middle Aged ; Prevalence ; Saliva ; Young Adult

OBJECTIVETo optimize formulas of Quban gel.

METHODThe U6 (6(2) x 3) uniform design was adopted to optimize gel formulas, with rheological parameters, such as viscosity and yield value in room temperature, viscosity and yield value in average temperature of skin, thixlotropy.

RESULTThe optimum proportion of matrix was made of 1.0 g carbomer 940, 5 mL glycerin and pH value 5-6.

CONCLUSIONThe regression model for gel matrix quality and gel rheological parameters was established to directly reflect the impacting effect of various factors, and provide certain preference basis for the screening of gel matrix formulas. Quban gel prepared by the method was evenly distributed, moderately viscous and highly thixotropic

Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Gels ; Quality Control ; Regression Analysis ; Viscosity

Objective To construct a recombinant baculovirus dual expression vector containing NIS gene under the control of human telomerase reverse transcriptase (hTERT) promoter and plasminogen kringle 5 (K5) gene driven by early growth response 1 (Egr1) promoter,and to explore the feasibility of targeting both tumor and tumor vessel with combination of radioiodide and antiangiogenic therapy.Methods The hTERT-NIS gene and Egr1-K5 gene fragments were subcloned into baculovirus vector,then packaged and amplified in the sf9 cells to obtain recombinant baculovirus Bac-hTERT-NIS-Egr1-K5.Bac-CMV-NISEgr1-K5,Bac-hTERT-0-Egr1-K5 and Bac-hTERT-NIS-Egr1-0 were constructed as controls.The expression of NIS and K5 genes in human cervix cancers cells (HeLa) was examined by Western blot and quantitative real-time PCR.Functional NIS activity was confirmed by the uptake of 125I,the inhibition of NaClO4 and the cytotoxicity of 131I.The apoptotic effect of 131I-inducedK5 on human umbilical veins endothelial cells (HUVEC)was analyzed by an apoptosis assay using flow cytometry.Statistical analysis was performed using the analysis of variance.Results The recombinant baculovirus Bac-hTERT-NIS-Egr1-K5 was successfully constructed.The NIS gene under the control of hTERT promoter was specifically expressed in HeLa cells.The baculovirusinfected HeLa cells showed a significant increase of 125I uptake,which was significantly inhibited by NaClO4(F199.296,P＜0.05).Furthermore,a notable decreased cell survival rate (38.3％) was found after 131I treatment.The expression of K5 gene induced by 131I was elevated in a dose or time dependent manner and resulted in obvious inhibition with cell survival rate of 30.8％ in baculovirus-infected HUVEC cells,which was significantly higher than that in the control groups (11.2％ and 10.9％ respectively,F=19.926,45.409;both P＜0.05).Conclusions A recombinant baculovirus dual expression vector containing the NIS and K5 genes has been successfully constructed.This study suggests the feasibility of a synergistic strategy of NISbased raidoiodide therapy and K5-based antiangiogenic therapy in vitro,and make it possible to perform in vivo study in the near future.

BACKGROUND: Representative formulae of Qi-Benefiting and Blood-Activating Chinese medicinal herbs Bu Yang Huan Wu Tang (BYHWT) can improve microcirculation and hypoxia, decrease fibrinogen, resist platelet aggregation, reduce blood lipid, dilate blood vessel, resist thrombosis, and reduce neonatal plaque rupture, and has a multi-target, multi-channel anti-atherosclerotic effect.
OBJECTIVE: To observe the influence of representative formulae of Qi-Benefiting and Blood-Activating Chinese medicinal herbs BYHWT on aorta Rho kinase, tissue factor, matrix metal oproteinase-2, matrix metal oproteinase-9 mRNA expression in atherosclerosis models, and to explore the action mechanism of BYHWT on anti-atherosclerosis.
METHODS: Vitamin D3 and high fat diet induced atherosclerosis in rats. 60 rats were randomly divided into normal control group, model group, 10 g/kg BYHWT therapy group, 20 g/kg BYHWT therapy group, simvastatin control group and BYHWT prevention group (n=10). At 28 days after model establishment, the expression level of Rho kinase, tissue factor, matrix metal oproteinase-2, matrix metal oproteinase-9 mRNA expression and blood lipid level were detected.
RESULTS AND CONCLUSION: (1) High fat diet and vitamin D3 could induce atherosclerosis in rats. Atherosclerotic plaque formed in the model group. (2) Rho kinase, tissue factor, matrix metal oproteinase-2, matrix metal oproteinase-9 mRNA expression and blood lipid were significantly lower in the normal control group, 20 g/kg BYHWT therapy group, simvastatin control group and BYHWT prevention group than in the model group (P < 0.01). Above indexes were significantly lower in the 10 g/kg BYHWT therapy group than in the model group (P < 0.05). No significant difference in above indexes was detected among BYHWT prevention group, 20 g/kg BYHWT therapy group and simvastatin control group (P > 0.05). (3) Results indicated that Qi-Benefiting and Blood-Activating BYHWT can resist atherosclerosis, down-regulate the expression of Rho kinase, tissue factor, matrix metal oproteinase-2, and matrix metal oproteinase-9 mRNA expression, which may be one of the mechanisms of anti-atherosclerosis.

OBJECTIVE:To optimize the preparation technology of Sinomenine topical paste. METHODS:Using“initial vis-cous force”,“holding viscous force”and“peeling strength”as index,heating and stirring time (A,h),heating temperature (B,℃) and the sequence of adding composition (softening agent,blank matrix and sinomenine)(C) as influential factors,the preparation technology of Sinomenine topical paste was optimized by orthogonal test and verified. At the same time,the content of sinomenine was determined by HPLC method. RESULTS:The optimal preparation technology of Sinomenine topical pasta was as follows as adding blank matrix and sinomenine,and then adding softening agent,heating at 80 ℃,stirring for 1 h. In verification test,RSD of comprehensive score for 3 batches of samples were 2.09%(n=3);average contents of samples were 6.7 mg/g, which was in line with the requirement of ≥6.0 mg/g. CONCLUSIONS:The optimal preparation technology of Sinomenine topical pasta is reasonable,stable and feasible. The paste shows good adhesiveness and is qualified in content.