2.Construction of prokaryotic expression vector, expression and purification of ginseng Cu/Zn superoxide dismutase.
Hong-Mei LIN ; Ze-Yu WANG ; Yue SHAO ; Xiao-Ye QIN ; Shi-Chao LIU ; Xin ZHANG ; Li-Min YANG
China Journal of Chinese Materia Medica 2013;38(23):4052-4055
The total RNA was extracted from ginseng leaves of Panax ginseng. The Cu/Zn-SOD gene was amplified via RT-PCR and the pET-28(a)-Cu/Zn-SOD expression vector was constructed. The pET-28 (a)-Cu/Zn-SOD recombinant plasmid was transformed into Escherichia coli BL21 (DE3) competent cells and was induced by IPTG in order to select optimal induction of expression conditions. The target protein was purified by the nickel ions (Ni ) affinity chromatography and the target protein enzyme activity was determinated by the xanthine oxidase method. The similarity of the Cu/Zn-SOD gene sequences and the Cu/Zn-SOD gene sequences of Korean ginseng in NCBI was 99. 00%. The target protein expression level was about 44.42%, and the molecular weight was 16.30 kDa after the pET-28(a)-Cu/Zn-SOD recombinants were induced by IPTG. The purified Cu/Zn-SOD protease activity reached 10,596.69 U x mg(-1). The P. ginseng pET-28(a)-Cu/Zn-SOD prokaryotic expression vector was built by the method of molecular biology, which provided the foundation for studying the Cu/Zn-SOD biology function.
Cloning, Molecular
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Escherichia coli
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genetics
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Gene Expression
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Genetic Engineering
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methods
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Genetic Vectors
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genetics
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Panax
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enzymology
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genetics
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Sequence Analysis
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Superoxide Dismutase
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genetics
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isolation & purification
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metabolism
3.Rapid simultaneous determination of ten major flavonoids in Tetrastigma hemsleyanum by UPLC-MS/MS.
Wen XU ; Zhi-Qin FU ; Jing LIN ; Xue-Cheng HUANG ; Hong-Min YU ; Ze-Hao HUANG ; Shi-Ming FAN
Acta Pharmaceutica Sinica 2014;49(12):1711-1717
In this study, a rapid and sensitive analytical method was developed for the determination of 10 major compounds (procyanidin B1, catechin, procyanidin B2, rutin, isoquercitrin, kaempferol-3-O-rutinoside, astragalin, quercitrin, quercetin, and kaempferol) in Tetrastigma hemsleyanum by using ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-MS/MS) in multiple-reaction monitoring (MRM) mode. UPLC-MS/MS assay with negative ion mode was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 μm) with the mobile phase consisting of acetonitrile (A) and 0.1% aqueous formic acid (B) in gradient elution at a flow rate of 0.25 mL · min(-1) and the column temperature was set at 45 °C. Under the optimized chromatographic conditions, good separation for 10 target compounds were obtained including chiral isomer procyanidins B1 and B2 were completely separated within 8.5 min. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r > 0.996 6), the overall recoveries were ranged from 95.44%-110.40% with the RSD ranging from 2.37%-8.69%. It is the first report about simultaneous analysis of 10 major flavonoids components in Tetrastigma hemsleyanum by using UPLC-MS/MS method, which affords highly sensitive, specific, speedy and efficient method for quality control of Tetrastigma hemsleyanum
Acetonitriles
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Chromatography, High Pressure Liquid
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Flavonoids
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chemistry
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Kaempferols
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Quercetin
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analogs & derivatives
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Rutin
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Tandem Mass Spectrometry
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Vitaceae
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chemistry
4.Analysis of angle Kappa variation in corneal refractive surgery under light and dark conditions
Rong, SHI ; Ze-Hong, DONG ; Wei, ZHAO ; Xue-Ting, CHEN ; Li-Rong, DONG ; Jie, FENG ; Yu-Sheng, WANG
International Eye Science 2016;16(8):1462-1464
?AIM:To observe the values and changing rules of angle Kappa in corneal refractive surgery under light and dark conditions.?METHODS:Two hundred and thirty-four eyes of 118 patients for corneal refractive surgery were enrolled for this study.Pupil diameters and angle Kappa values under light and dark conditions were measured by Keratron Scout corneal topography.?RESULTS: There were significant differences in pupil diameters between light and dark conditions (P<0.01). More angle Kappa of both eyes distributed in the superior nasal quadrant under light conditions, and more angle Kappa distributed in the superior temporal quadrant under dark conditions.The differences of horizontal and vertical offsets of angle Kappa under two conditions were statistically significant (P<0.01).?CONCLUSION: The changes of pupil diameters in light and dark conditions could affect angle Kappa and then affect the accuracy of corneal refractive surgery centered on angle Kappa.
5.The effects of nucleoside analogues on hepatitis B virus in hepatic lymph nodes of hepatitis B virus-associated liver transplantation recipients.
Ze CHEN ; Hong ZHENG ; Rui SHI ; Wei-ping ZHENG ; Zhi-qiang CHU ; Hong-li SONG ; Zhong-yang SHEN
Chinese Journal of Surgery 2012;50(9):835-838
OBJECTIVETo investigate the effects of nucleoside analogues on hepatitis B virus (HBV) in hepatic lymph nodes of hepatitis B related liver transplantation recipients who were hepatitis B surface antigen (HBsAg) positive but negative for serum HBV DNA.
METHODSFrom June 2010 to March 2011, thirty-six cases of hepatitis B related liver transplantation recipients [32 males, 4 females, average age (54 ± 7) years] were divided into drug treatment group and non-drug treatment group according to the utility of nucleoside analogues. Drug treatment group was divided into two subgroups: drug treatment > 3 months group and drug treatment ≤ 3 months group. The hepatic lymph nodes in the hepatoduodenal ligament were taken during the operation of liver transplant. Using nested or semi-nested PCR, HBV DNA and the replicative form HBV cccDNA in hepatic lymph nodes were detected. Data were analyzed by Fisher's exact test.
RESULTSThe positive rate of HBV DNA: the difference was not statistically significant between drug treatment group (72.7%, 16/22) and non-drug treatment group (14/14) (P = 0.062), the difference was not statistically significant between drug treatment > 3 months group (10/14) and drug treatment ≤ 3 months group (6/8) in the subgroups of drug treatment group (P = 1.000). The positive rate of HBV cccDNA: drug treatment group (22.7%, 5/22) was significantly lower than the non-drug treatment (12/14) (P = 0.000), drug treatment > 3 months group (1/14) was significantly lower than drug treatment ≤ 3 months group (4/8) in the subgroups of drug treatment group (P = 0.039).
CONCLUSIONSHepatic lymph nodes maybe one of the extrahepatic HBV reservoirs. Treating with nucleoside analogues more than 3 months can significantly decrease the replication of HBV in hepatic lymph nodes of HBV associated liver transplantation recipients.
Adult ; Aged ; DNA, Viral ; analysis ; Female ; Hepatitis B ; drug therapy ; Hepatitis B virus ; drug effects ; physiology ; Humans ; Liver Transplantation ; Lymph Nodes ; virology ; Male ; Middle Aged ; Nucleosides ; therapeutic use ; Preoperative Care ; Virus Replication
6.Perioperative management of modified uvulopalatopharyngoplasty.
Zhi-hong LUO ; Shi-ming CHEN ; Ze-zhang TAO ; Yong-mao CAO
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2006;41(2):100-103
OBJECTIVEExperiences and lessons of uvulopalatopharyngoplasty (UPPP ) perioperative management, especially causes of postoperative tracheotomy, were analyzed, and related strategy was raised to have a better perioperative management and to avoid tracheotomy.
METHODSTwo hundred and fifty eight cases of obstructive sleep apnea hypopnea syndromes (OSAHS) diagnosed with polysomnography (PSG) were treated with modified uvulopalatopharyngoplasty (UPPP). The perioperative management was summarized. Patients were divided into two groups according to the perioperative management: without or with perioperative comprehensive management. In group A, there were 32 patients, without comprehensive management, and in group B there were 226 cases with comprehensive management. Sixty eight cases in group B whose apnea hypopnea index over 50 times per hour and the lowest arterial oxygen saturation was less than 0.5 were treated with continuous positive airway pressure (CPAP) for 1 to 3 weeks. For all the 258 cases, perioperative management includes treatment of medical complications, treatment with antibiotics 2 or 3 days before the operation. None of these cases had tracheotomy before surgery.
RESULTSIn group A, three of 32 patients had postoperative tracheotomy, two because of bleeding, and another one because of laryngeal spasm. In group B, none of 226 patients underwent tracheotomy, which owing to modified operative apparatus and effective perioperative and postoperative treatment (chi2 = 21.35, P < 0.001). In group A, 5 of 32 patients had oral pharynx bleeding after 24 hours of the operation. While 26 of 226 patients in group B did so (chi2 = 0.15, P > 0.05).
CONCLUSIONComprehensive perioperative management can effectively lower down the complication rate for patients receiving uvulopalatopharyngoplasty.
Adult ; Cleft Palate ; surgery ; Female ; Humans ; Male ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; adverse effects ; Palate ; surgery ; Pharynx ; surgery ; Postoperative Complications ; surgery ; Retrospective Studies ; Sleep Apnea, Obstructive ; surgery ; Tracheotomy ; adverse effects ; Treatment Outcome ; Uvula ; surgery
7.Clinical study of method of strengthening body resistance and disintoxication disintoxication in patients with HCC of post-TACE.
Ze-xiong CHEN ; Shi-jun ZHANG ; Hong-tao HU ; Bao-guo SUN ; Li-rong YIN
China Journal of Chinese Materia Medica 2007;32(12):1211-1213
OBJECTIVETo research the effect of a complex prescription of Chinese crude drug with the function of strengthening body resistance and disintoxication disintoxication in patients with HCC of post-TACE.
METHOD45 patients with HCC of post-TACE, as the treatment group, were treated by a complex prescription of Chinese crude drug with the function of strengthening body resistance and disintoxication disintoxication and routine methods of protecting liver. Other 37 patients, as the control group, with the same clinical feature were treated by routine methods of protecting liver only. In the later 1 month, accumulated points of clinical symptom, hepatic function and AFP were observed in all of the patients. And the clinical effect of the two groups was compared.
RESULTOne week later, in the treatment group, there is no improvement in anorexia but nausea, abdominal distention and lassitude were improved more obviously than pretherapy in both a week and one month later (P < 0.01 or P <0.05). In the control group, anorexia were improved a week later (P <0.05), but there is no improvement in nausea, abdominal distention and lassitude at the same time, and one month later all of the indexes above improved (P <0.01 or P <0.05). Accumulated points of clinical symptom was decreased more obviously in the treatment group than in the control group in both a week and one month later (P <0.05). At the end of the therapy, in the both groups, ALT, TBIL and AFP all improved except ALB, (P <0.01 or P <0.05). And TBIL improved more obviously in the treatment group than in the control one month later (P <0.05).
CONCLUSIONThis complex prescription of Chinese crude drug can lighten the adverse reaction of post-TACE. And also it can promote the recovery of liver function and evaluate the quality of lives of such patients.
Anorexia ; drug therapy ; etiology ; Carcinoma, Hepatocellular ; therapy ; Chemoembolization, Therapeutic ; adverse effects ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Fatigue ; drug therapy ; etiology ; Female ; Humans ; Liver Function Tests ; Liver Neoplasms ; therapy ; Male ; Nausea ; drug therapy ; etiology ; Phytotherapy ; Plants, Medicinal ; chemistry ; Treatment Outcome ; alpha-Fetoproteins ; metabolism
8.Expression of PTEN in athymic mice with HCC treated by complex prescription of Chinese crude drug.
Bao-guo SUN ; Ze-xiong CHEN ; Shi-jun ZHANG ; Yong-dong LIU ; Hong-zhong HUANG ; Li-rong YIN
China Journal of Chinese Materia Medica 2007;32(11):1057-1060
OBJECTIVETo research the treatment effect of complex prescription of Chinese crude drug in BALB/c athymic mice with human liver cancer, which were built by Bel-7402.
METHOD48 male BALB/c athymic mouse models were built by Bel-7402 with an indirect method. After 24 hours of postoperation, the 48 athymic mice were distributed randomly into 4 groups which were treated by intragastric administration with complex prescription of Chinese crude drug that had been deliquated into 3 groups by the different density as the low, middle, and high and FT207 (Tegafur) for 4 weeks. At last, athymic mice were put to death and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry (PowerVision Two-Step Histostaining Reagent).
RESULTAll of the 48 athymic mice survived 12 to 28 days (Ms 24 days) and every mouse with liver cancer demonstrated by dissection. The result of immunohistochemistry represents that the intensity of PTEN in latero-cancer tissue is the highest, and then the hepatic tissue, the lowest is cancer tissue, P < 0.01. It also represents that the intensity of PTEN in treatment groups (A, B, C) is more higher than the control group (D), P < 0.05 or P < 0.01, and group B is the highest in the treatment groups, P < 0.05 or P < 0.01. However, there is no significant statistic difference between group A and group C.
CONCLUSIONThe higher expression of PTEN in the laterocancer tissue can represent the protective reaction of stress of the organism. And anticancer effect of this complex prescription of Chinese crude drug relates to an eligible density of it. Mechanisms of this complex prescription of Chinese crude drug healing HCC may partially be explained by enhancing the expression of PTEN in liver.
Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; enzymology ; pathology ; Cell Line, Tumor ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Humans ; Immunohistochemistry ; Liver ; drug effects ; enzymology ; pathology ; Liver Neoplasms ; drug therapy ; enzymology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; PTEN Phosphohydrolase ; metabolism ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Xenograft Model Antitumor Assays
9.Mitochondria dysfunction was involved in copper-induced toxicity in MES23.5 cells.
Li-Min SHI ; Hong JIANG ; Jun WANG ; Ze-Gang MA ; Jun-Xia XIE
Neuroscience Bulletin 2008;24(2):79-83
OBJECTIVETo investigate the toxicity of copper on MES23.5 dopaminergic cells and the probable mechanisms involved in this process.
METHODSMES23.5 dopaminergic cells were selected as our experimental model. [3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide] (MTT) assay was used to detect the influence of copper on the cell viability. The semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), Western blotting and the high performance liquid chromatography-electrochemical detection (HPLC-ECD) have been used to detect the tyrosine hydroxlase (TH) mRNA and protein expression and the dopamine content in MES23.5 cells. The flow cytometry have been used to detect the changes of mitochondrial transmembrane potential.
RESULTS100 and 200 mumol/L copper had no effect on the MES23.5 cell viability, whereas 400 and 800 mumol/L of copper could decrease the cell viability (P < 0.01). Treating cells with 200 mumol/L copper for 24 h decreased the TH mRNA expression, the TH expression and the dopamine content compared with the control (P < 0.01, P < 0.01, P < 0.05, respectively). Besides, the mitochondrial transmembrane potential also decreased with the treatment of 200 mumol/L copper for 24 h (P < 0.01).
CONCLUSIONCopper could exert the toxic effects on MES23.5 dopaminergic cells and decrease the cell function. The dysfunction of mitochondria may be the mechanism of this toxicity effect.
Animals ; Cell Survival ; drug effects ; genetics ; Cells, Cultured ; Copper ; metabolism ; toxicity ; Dopamine ; biosynthesis ; Dose-Response Relationship, Drug ; Hybridomas ; Membrane Potential, Mitochondrial ; drug effects ; genetics ; Mice ; Mitochondria ; drug effects ; metabolism ; pathology ; Nerve Degeneration ; chemically induced ; metabolism ; physiopathology ; Neurons ; drug effects ; metabolism ; pathology ; Neurotoxins ; toxicity ; Oxidative Stress ; drug effects ; physiology ; Parkinson Disease ; etiology ; metabolism ; physiopathology ; RNA, Messenger ; drug effects ; metabolism ; Rats ; Tyrosine 3-Monooxygenase ; drug effects ; genetics ; metabolism
10.The level of welding fume and the healthy status of dust workers in electric welder's pneumoconiosis surveillance sentinel of one city.
Chun-hua LU ; Bao-li ZHU ; Ji-hong YANG ; Bang-mei DING ; Ze-yun YANG ; Ping ZHOU ; Shi-wei YIN ; Li-zhuang XIE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2013;31(11):847-848