2.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
3.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
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.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
7.Unilateral buccinator myomucosal island flap with double opposing Z-plasty for wider palatal cleft repair.
Ze-hong SHI ; Sen-kai LI ; Yang-qun LI ; Ning-bei YIN ; Bin XIONG ; Zhen-min ZHAO
Chinese Journal of Plastic Surgery 2007;23(4):290-292
OBJECTIVETo explore a method to repair larger cleft palate and lengthen soft palate without oral palate raw surface and scar formation, reduce the effect on maxilla and dental arch development.
METHODSA modified double opposing Z-plasty was used to lengthen soft palate and the nasal palate was closed by using large turn-over mucoperiosteal flaps on the oral surface of the junction of the hard palate and soft palate, oral raw surface on the palate was closed by a buccal myomucosal island flap.
RESULTSThirty-six palates have been repaired by this procedure, all of which had satisfactory results without flap necrosis, infection, difficulties in opening mouth and facial nerve injury except two post-operative fistulas. Eight patients were followed up and all display complete velopharyngeal closure.
CONCLUSIONSUsing unilateral buccinator myomucosal island flap with double opposing Z-plasty to repair wider palatal cleft can get a satisfactory soft palate lengthening. At the same time it can avoid bone surface exposing and scar formation; it is a safe and reliable procedure.
Adolescent ; Cheek ; surgery ; Child ; Child, Preschool ; Cleft Palate ; surgery ; Female ; Humans ; Male ; Mouth Mucosa ; transplantation ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Young Adult
8.Qualitative and quantitative analysis of major constituents in Tetrastigma hemsleyanum by HPLC-Q-TOF-MS and UPLC-QqQ-MS.
Wen XU ; Zhi-qin FU ; Jing LIN ; Xue-cheng HUANG ; Dan CHEN ; Hong-min YU ; Ze-hao HUANG ; Shi-ming FAN
China Journal of Chinese Materia Medica 2014;39(22):4365-4372
A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS) was developed for identification of multi-constituents and an analytical method was developed for simultaneously determining 4 major compounds (rutin, isoquercitrin, kaempferol-3-0-rutinoside, and astragalin) in Tetrastigma hemsleyanum Diels et Gilg. The HPLC-Q-TOF-MS assay was performed on a Welch Ultimate XB-C18 column (4.6 mm x 150 mm, 5 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% Formic acid (B) in gradient mode at a flow rate of 0.8 mL x min(-1). The column temperature was at 30 degrees C, and negative ion mode was used for TOF-MS. The UPLC-QqQ-MS assay was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% formic acid (B) in gradient mode at a flow rate of 0.25 mL x min(-1). The column temperature was at 45 degrees C, and MRM mode was used for QqQ-MS. Based on the retention time and MS spectra, 24 compounds were identified or tentatively characterized by comparing with reference substances or literatures. For quantitative the linear range of 4 detected compounds were good (r > 0.9966), and the overall recoveries ranged from 98.27% to 101.58%, with the RSD ranging from 3.15% to 5.88%. The results indicated that new approach conbined HPLC-Q-TOF-MS and UPLC-QqQ-MS was applicable in qualitative and quantitative quality control of Tetrastigma hemsleyanum.
Acetonitriles
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chemistry
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Chromatography, High Pressure Liquid
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methods
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Formates
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chemistry
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Tandem Mass Spectrometry
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methods
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Vitaceae
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chemistry
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Water
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chemistry
9.An improved imaging analysis for quantitative measurement of brain slice volume.
Ze-hua LIANG ; Er-qing WEI ; Chao-yang ZHU ; Shi-hong ZHANG ; Hui-min XU
Journal of Zhejiang University. Medical sciences 2003;32(6):497-501
OBJECTIVETo improve computer-assisted imaging analysis for quantitatively measuring brain slice volume of rats and mice in comparison with conventional measuring methods,and to evaluate its usefulness in assessment of focal cerebral ischemia.
METHODSThe accurate volumes of rat and mouse brain slices were measured by weight and special gravity measuring. The areas of brain slices were measured by imaging analysis, then the slice volumes of right and left hemispheres were calculated by multiplying the adjusted thickness of the slices. In addition, the brain slice volumes of right and left hemispheres from focal cerebral ischemic mice were compared to assess ischemic injury using the imaging analysis.
RESULTArea measurement by computer-assisted imaging analysis was linear with different accurate areas (r=1.000). Slice volumes measured by imaging analysis correlated well with the accurate volumes measured by special gravity method, r=0.809 (n=45, P<0.001) in rats, and r=0.844 (n=74, P<0.001) in mice. The brain volumes in ischemic hemispheres were larger than in non-ischemic hemispheres in ischemic mice.
CONCLUSIONComputer-assisted imaging analysis can measure the brain slice volumes accurately and compare right and left hemisphere volumes quantitatively.
Animals ; Brain ; pathology ; Brain Ischemia ; pathology ; Image Processing, Computer-Assisted ; Male ; Mice ; Mice, Inbred ICR ; Rats ; Rats, Sprague-Dawley
10.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