3.Simultaneous determination of six Salvia miltiorrhiza gradients in rat plasma and brain by LC-MS/MS.
Sheng-Min LIU ; Zhi-Hong YANG ; Xiao-Bo SUN
China Journal of Chinese Materia Medica 2014;39(9):1704-1708
To develop a LC-MS/MS method for the determination of protocatechuic acid, protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, cryptotanshinone and tanshinone II(A) in rat plasma and brain. The plasma and brain samples were precipitated with ethyl acetate, then were separated on an Agilent eclipse plus-C18 column (2.1 mm x 50 mm, 3.5 microm) using acetonitrile (consisting of 0.1% formic acid) and water (consisting of 0.1% formic acid) as mobile phase in gradient elution mode. The mass spectrometer was operated under both positive and negative ion mode with the ESI source, and the detection was performed by MRM. The transition of 154.3/153.1 m/z for protocatechuic acid, 137.3/108 m/z for protocatechuic aldehyde, 493.0/295.2 m/z for Salvianolic acid A, 718.0/520.0 m/z for salvianolic acid B, 321.4/152.3 m/z for chloramphenicol, 297.4/254.3 m/z for cryptotanshinone, 295.5/249.3 m/z for tanshinone II(A) and 285.2/154.0 m/z for Diazepam. The calibration curves in the range of 0.625-1 000 microg x L(-1) for protocatechuic acid and protocatechuic aldehyde, 1.25-1 000 microg x L(-1) for salvianolic acid A, 2.5-1 000 microg x L(-1) for salvianolic acid B, 0.15-1 000 microg x L(-1) for cryptotanshinone, 0.625-1 000 microg x L(-1) for tanshinone II(A) are with good linearityin rat plasma and brain. The analysis method is sensitive, simple, and suitable enough to be applied in the pharmacokinetic study of the 6 main components. Animal testing gives the lgBB of the drugs and further studies of the 6 components cross the blood-brain barrier can be carried out.
Animals
;
Benzaldehydes
;
administration & dosage
;
blood
;
pharmacokinetics
;
Benzofurans
;
administration & dosage
;
blood
;
pharmacokinetics
;
Blood-Brain Barrier
;
metabolism
;
Brain
;
metabolism
;
Caffeic Acids
;
administration & dosage
;
blood
;
pharmacokinetics
;
Catechols
;
administration & dosage
;
blood
;
pharmacokinetics
;
Chromatography, Liquid
;
methods
;
Diterpenes, Abietane
;
administration & dosage
;
blood
;
pharmacokinetics
;
Hydroxybenzoates
;
administration & dosage
;
blood
;
pharmacokinetics
;
Injections, Intravenous
;
Lactates
;
administration & dosage
;
blood
;
pharmacokinetics
;
Phenanthrenes
;
administration & dosage
;
blood
;
pharmacokinetics
;
Plant Preparations
;
administration & dosage
;
blood
;
pharmacokinetics
;
Rats
;
Reproducibility of Results
;
Salvia miltiorrhiza
;
chemistry
;
Tandem Mass Spectrometry
;
methods
4.Intra-operative implantation of I125 seeds to treat advanced pancreatic cancer
Yang LIU ; Bo SHENG ; Kai JIANG ; Yanbin WANG
Chinese Journal of Hepatobiliary Surgery 2014;20(6):446-448
Objective To study the clinical efficacy of intra-operative implantation of I125 seeds to treat advanced pancreatic cancer.Methods We retrospectively analyzed the clinical data of 28 patients who underwent intra-operative implantation of I125 between January 2009 and June 2010 to treat advanced pancreatic cancer.Patients who received conservative treatment (n =136) at the same period were used as the control group.Results The mean tumor diameter was (5.7 ± 1.7) cm.The average number of implanted seeds were 50.There were no patients who suffered from pancreatic fistula or post-operative bleeding,and no patient died from the treatment.Twenty-five patients were regularly followed up,and the follow-up rate was 85.7%.The survival rates at one month,half a year and one year were 100%,70.8% and 26.9%,respectively.The average survival time was (9.9 ± 1.4) months.The survival data were significantly higher than that in the control group.Conclusion Intra-operative implantation of I125 seeds was an efficacious treatment for patients with advanced pancreatic cancer.
5.Research of Desulfhydrase Involved in L-cysteine Biosynthetic Pathway in Pseudomonas sp.
Yang-Sheng YU ; Yang LI ; Yong-Jie JIN ; Gang BAI ; Wen-Bo YANG ;
Microbiology 1992;0(04):-
The L-cysteine desulfhydrase gene (cd) of Pseudomonas sp.TS1138 was amplified by PCR,and the amplified gene was recombined in the cloning vector pBluescript SKII.The 1.2kb DNA fragment containing cd was sequenced,and its homology with other desulfhydrases was blast; then the cd was cloned into the expression vector pET-21a(+), and afterward expressed by IPTG inducement.The expression protein was purified by Ni-NTA His-Bind Resin.Then the expression protein was identified by the method of activity staining of desulfhydrase, and the characterization of L-cysteine desulfhydrase and the critical role it played in the L-cysteine biosynthetic pathway were discussed.
6.Purification and Properties of L-Cysteine Synthetase from Pseudomonas
Yong-Jie JIN ; Wen-Bo YANG ; Zhong LIU ; Gang BAI ; Yang-Sheng YU ;
Microbiology 1992;0(06):-
The L-ATC hydrolase and L-SCC amidohydrolase which convert L-ATC to L-cysteine in Pseudomonas sp.TS-1138 are purified about 83.9 and 90.3 fold by salting-out method, Sephadex G-75 gel chromatography, DEAE-cellulose 52 ion exchange and Sephadex G-100 gel chromatography, etc. The purified enzyemes are both demonstrated by SDS-PAGE to be a homogeneous protein. Their molecular weight are about 37.5kD and42.8kDa respectively. The optimum reaction temperature are both 35℃, and the optimum pH are 7.0 and 8.0 respectively. The Km of the two enzymes are 0.67 mmol/L and 0.15 mmol/L, and the Vmax are 0.39?10 -3mmol/L?min and 0.42?10 -3mmol/L?min respectively.
7.Dynamic Changes of Phenotypes and Secretory Functions during the Differentiation of Pre-DCs to Mature DCs
WANG JING ; ZHAO LI-BO ; CHANG SHENG ; MING CHANG-SHENG ; YANG JUN ; GONG NIAN-QIAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(2):191-196
The dynamic expression of cytokines and phenotypes during the differentiation process of dendritic cell precursors (pre-DCs) to mature dendritic cells (DCs) was investigated,and the effects of inflammatory stimulation with lipopolysaccharide (LPS) on DCs differentiation were understood.The differentiation of bone marrow cells isolated from Balb/c mice was induced to DCs in an 8-day cell culture system with RPMI-1640 complete culture medium containing 10% FBS,20 ng/mL recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and 10 ng/mL recombinant mouse interleukin-4 (rmIL-4).On the day 3,6 and 7 after culture,DCs were divided into non-LPS group and LPS group,given 500 ng/mL LPS for 24 h stimulation and no stimulation respectively.The expression levels of CD 1 l c+,MHC-Ⅱ +,CD40+,CD80+ and CD86+ were detected by flow cytometry,and those of IL-2,IL-4,IL-10,IL-12 p70 and IFN-γ in the supernatant by ELISA.On the day 3 and 6 after culture,the expression of IL-2,IL-4,IL-10 and IFN-γ in DCs showed no significant differences between non-LPS group and LPS group,whereas the differences were significant at day 7.The expression levels of cytokines (for IL-2,IL-4,IL-10,IFN-γ and IL-12 p70:152.86±6.91,778.33±8.35,44.55±2.54,5.8.26±1.09 and 2423.00±57.21 pg/mL respectively) in LPS group were higher than those in non-LPS group,especially IL-12 p70 increased obviously at day 7.It was concluded that during the differentiation process of pre-DCs to mature DCs,LPS stimulates DCs producing large amounts of IL-12 p70 and Thl-type cytokines as compared with Th2-type cytokines,and day 7 may be a key time-point for DCs polarization.
8.Analysis of surveillance results of drinking-water supply status in drinking-water-borne endemic fluorosis areas in Henan
Bo, YU ; Li, ZHANG ; Guo-qiang, HOU ; Yang, LIU ; Chun-sheng, YUAN ; Gan, CHEN
Chinese Journal of Endemiology 2013;(2):186-188
Objective To understand the status of drinking-water supply and the progress of waterimproving projects in drinking-water-borne endemic fluorosis areas in Henan,and to provide scientific data for related government departments to carry out prevention and surveillance plan for those areas.Methods Questionnaire survey was carried out in all the villages in drinking-water-borne endemic fluorosis areas in Henan in 2010.Each village was given one set of questionnaire.Results By the end of 2010,the total number of fluorosis villages in Henan was 25 434,among them 11 484 villages had been conducted water-improving projects,accounting for 45.15%.Currently,9267 water-improving projects worked properly,accounting for 36.4%(9267/25 434)of all villages surveyed,and 80.7% (9267/11 484)villages had water-improving projects.The projects in 2217 villages do not work properly,accounting for 19.3%(2217/11 484) of all projects.There were 5832 water-imp.roving projects conducted amnog 2005-2010,accounting for 50.8% (5832/11 484) of all water-improving projects.Among villages with no such projects,97.5%(15 769/16 167) of them used shallow groundwater,and in villages with water-improving projects,89.6% (8303/9267) of them used deep groundwater as drinking-water supply.Conclusions After 2005,the water improving progress was significantly speed up in drinking-water-borne endemic fluorosis areas in Henan,but progress of the water improving projects is relatively slow due to changing and expanding of fluorosis areas.The government should increase investment and improve the proportion of waterimproving defluoridation.
9.Combination chemotherapy with etoposide and cisplatin for high-risk,chemorefractory and recurrent gestational trophoblastic neoplasia
Jie JIANG ; Fang-Fang NAN ; Xing-Sheng YANG ; You-Zhong ZHANG ; Bo WANG ; Bei-Hua KONG ;
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
Objective To evaluate the feasibility and effectiveness of combination chemotherapy with etoposide and cisplatin(EP)regimen on the patients with high-risk,chemorefractory and recurrent gestational trophoblastic neoplasia(GTN).Methods Thirty-nine patients with gestational trophoblastic tumors were analyzed retrospectively,25 of 39 patients were of high-risk,9 patients were chemorefractory and 5 patients were recurrent.All 39 patients were administrated with EP regimen,and 10 patients were assisted with surgery.All the patients were followed up.Clinical response,toxicity,the occurrence of secondary tumors of all patients,and the fertility of 30 patients whose fertility function was preserved were investigated. Results Thirty-nine GTN patients underwent a total of 221 cycles of the EP regimen.The average number of courses for each patient was 5.7.The total complete remission rate of the regimen was 74%(29/39). Twenty-five patients with high-risk GTN received a total of 139 cycles and the average number of courses was 5.6.Nineteen patients achieved complete remission and 6 patients showed drug-resistant.The complete remission rate of the high-risk group was 76%(19/25).Nine patients with chemorefractory GTN obtained a total of 55 cycles and the average number of courses was 6.1.Six patients achieved complete remission and 3 patients showed drug-resistant again.The complete remission rate of the chemorefractory group was 6/9. Five patients with recurrent GTN received 27 cycles and the average number of courses was 5.4.Four patients achieved complete remission,1 patient showed drug-resistance and died.Bone marrow toxicity, gastrointestinal reaction and alopecia were the main side effects of the EP regimen,but the bone marrow toxicity was slight and no grade Ⅳ side effect occurred.No fatal effect was found.Eight of 30 patients whose fertility faction was preserved had become pregnant after recovery,with a total of 8 pregnancies.Among them,2 were terminated by induced abortion,and 6 underwent normal term delivery and gained 6 infants who had no congenital malformation.All the 6 children had normal growth and development after childbirth. None of the women developed secondary tumors.Conclusion The EP regimen is effective and safe for the treatment of high-risk,chemorefractory and recurrent GTN.
10.Investigation of the carotid intima-media thickness in 221 individuals with metabolic syndrome
Wen-Sheng JIN ; Chang-Yu PAN ; Ju-Ming LU ; Guang ZHI ; Bo YANG ;
Chinese Journal of Endocrinology and Metabolism 1986;0(03):-
Metabolic abnormalities were identified and carotid intima-media-thickness(IMT)was measured in 221 individuals at risk for metabolic syndrome(MS).The results indicated that IMT was significantly thicker in MS individuals than that in non-MS individuals(P<0.01).And there was a tendency of progressive increase in IMT with increasing components of metabolic syndrome.