1.A novel lupane triterpene from seeds of Ziziphus jujuba var.spinosa
Chinese Traditional and Herbal Drugs 1994;0(02):-
Objective To study the chemical constituents from the seeds of Ziziphus jujuba var.spinosa,which is used as a sedative herbal medicine in China.Methods The constituents were separated and purified by silica gel column.Their structures were elucidated based on their physicochemical properties and spectral analysis.Results Six compounds were obtained from 95% EtOH extracts in the seeds of Z.jujuba var.spinosa.They were identified as 2?,3?-dihydroxy-lup-20(29)-en-28-oic acid methyl ester(Ⅰ),?-sitosterol(Ⅱ),betulin(Ⅲ),betulinic acid(Ⅳ),hexadexanoic acid 2,3-dihydroxypropyl ester(Ⅴ),and daucosterol(Ⅵ).Conclusion Compound Ⅰ is a novel lupane triterpene named as alphitolic acid methyl ester,compound Ⅴ is obtained from the seeds of Z.jujuba var.spinosa for the first time.
2.Bladder Cancer J82 Cells Apoptosis Induced by Brucea Javanica Oil Emulsion and the Mechanism
Feng LV ; He WANG ; Weijun QIN
Chinese Journal of Rehabilitation Theory and Practice 2007;13(5):432-433
Objective To study the effect of Brucea javanica oil emulsion on apoptosis of bladder cancer J82 cells and the possible mechanism.Methods Brucea javanica oil emulsion of different concentrations (0.5~4.0 ml/L) was added to the bladder cancer J82 cell cultured in vitro.After 24 h, the proliferation of J82 cells was measured by methyl thiazole tetrazolium (MTT) assay, cell apoptosis was detected by morphological observation and flow cytometer analysis, and caspase-3 activity was assessed using a caspase-3 kit. Results 0.5~4.0 ml/L Brucea javanica oil emulsion inhibited J82 cell proliferation in a dose-dependant manner.2.0 ml/L Brucea javanica oil emulsion induced typical apoptosis 24 h after incubation. The caspase-3 activity of 2.0 ml/L Brucea javanica oil emulsion group was higher than those of control group or Brucea javanica oil emulsion plus Ac-DEVD-CHO group. Conclusion Brucea javanica oil emulsion induces J82 cell apoptosis, and this effect includes activation of caspase-3.
3.LC-MS/MS method for quantification and pharmacokinetic study of gabapentin in human plasma.
Zhili XIONG ; Jia YU ; Jifen HE ; Feng QIN ; Famei LI
Acta Pharmaceutica Sinica 2011;46(10):1246-50
A sensitive, rapid and specific liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for quantification of gabapentin in human plasma has been developed. After a single plasma protein precipitation with methanol, gabapentin and metformin (internal standard) were chromatographed on a Inertsil ODS-3 column (50 mm x 2.1 mm ID, 3 microm) with mobile phase consisting of methanol-0.2% formic acid aqueous solution (80:20, v/v) at a flow-rate of 0.2 mL x min(-1). Electrospray ionization (ESI) source was applied and operated in the positive ion mode. Multiple reaction monitoring (MRM) mode with the transitions of m/z 172 --> m/z 154 and m/z 130 --> m/z 71 were used to quantify gabapentin and metformin, respectively. The run time was 2.2 min. The linear calibration curve was obtained in the concentration range of 40.8-8.16x10(3) ng x mL(-1). The lower limit of quantification was 40.8 ng x mL(-1). The intra- and inter-day precision (RSD) was less than 12%, and the accuracy (RE) was within +/-6.4% calculated from quality control (QC) samples. The method was used to determine the concentration of gabapentin in human plasma after a single oral administration of 600 mg gabapentin capsule to 20 healthy male Chinese volunteers. The method was proved to be selective, sensitive, rapid and suitable for pharmacokinetic study of gabapentin in human plasma.
5.Effects of different degrees of intermittent hypoxia on NF-κB, IL-10 and visfatin in 3T3-L1 adipocytes
Miaomiao HAN ; Qin ZHOU ; Jing FENG ; Wenyan NIU ; Qing HE
Tianjin Medical Journal 2016;44(9):1124-1127
Objective To determine levels of nuclear factor (NF)-κB, interleukin (IL)-10, and visfatin in adipocytes treated by different degrees of intermittent hypoxia (IH), and to investigate the mechanism of IH leading to insulin resistance (IR). Methods The cell model of intermittent hypoxia/re-oxygenation (IH/ROX) in obstructive sleep apnea (OSA) was established. Differentiation mature 3T3-L1 adipocytes, were randomly divided into 10 groups including four different-frequency intermittent hypoxia groups(IH1-4, fixed intermittent hypoxia scheme for 1.5%O2 45 s and then re-oxygen 21%O2 for 2 min 15 s, 4 min 15 s, 5 min 45 s and 8 min 45 s, 60 times circulation), and their normal oxygen control groups (SC1-4, instead each IH group 1.5%O2 to 21%O2, the rest groups were treated as same as IH group), continuous hypoxia group (CH, 10%O2 for 6 h) and normal oxygen control group (CC, 21%O2 for 6 h). ELISA method was used to determine the levels of IL-10 and visfatin in the supematant of adipocytes. Western blot method was used to determine the protein levels of NF-κB p65 and visfatin. Real-time PCR method was used to determine the mRNA levels of IL-10 and visfatin. Results The protein and mRNA expressions of IL-10 were significantly lower in IH group and CH group than those of control groups (P<0.01). The levels of NF-κB p65 protein were significantly increased in IH group and CH group than those of control group. The protein and mRNA expressions of visfatin were significantly higher in IH1, IH2 and CH groups than those of control group (P<0.01). Conclusion As a prominent feature of OSA pathophysiology, IH may take part in insulin resistance of OSA patients by abnormally secreting NF-κB, IL-10 and visfatin in adipocytes.
6.Immunological regulation mechanism of (-)-epigallocatechin-3-gallate on rheumatoid arthritis
Qin DING ; Hong NIE ; Dongyi HE ; Wei FENG
Chinese Journal of Rheumatology 2011;15(8):526-530
Objective To study the (-)-epigallocatechin-3-gallate (EGCG) function on the proliferation of T cells derived from the peripheral blood and synovial fluid of rheumatoid arthritis (RA) and RA-related cytokine levels and the role of EGCG on RA synovial fibroblasts (FLS) proliferation was investigated. Methods ① Mononuclear cells from RA peripheral blood (30 cases) and synovial fluid (23 cases)were isolated. Blank group, negative control group, positive control methotrexate (MTX) group and therapeutic group with three different concentrations of EGCG were set up. Incorporated isotope 3H was used to test T cell proliferation from RA-PBMC and SFMC. ELISA assay was used to test cytokine (TNF-α, IFN-γ, IL-1, IL-6 and IL-17A) levels. ② MTT assay was used to test FLS proliferation from RA synovial tissue (8 cases).Results ① The CPM value of the high-dose group of EGCG in the peripheral blood and synovial fluid of RA patients was [ ( 15 136±2910), ( 11 587±3135 ) ], which was declined significantly than the control group (42856±2127) (P<0.01). The levels of TNF-α, IFN-γ, IL-1, IL-6 and IL-17A in the high-dose group of EGCG in the peripheral blood were [(321±13), (298±20), (132±12), (197±7), (59±8) pg/ml], which were decreased significantly than those of the control group [ (458±28), (505±26), (346±28), (405±25),(109±13) pg/ml ] (P<0.05 or P<0.01 ). The levels of TNF-o, IFN-γ, IL-1, IL-6 and IL-17A in the highdose group of EGCG in the synovial fluid were [(41.4±2.9), (182±16), (56.3±11.0), (34.2±1.9), (44±8)pg/ml ], which was decre-ased significantly than the control group [ ( 388.3± 19.3 ), (469±20), ( 104.2±17.8 ),( 114.5±4.8), ( 104±11 ) pg/ml] (P<0.05 or P<0.01 ). ② The level (A) of the high-dose group of EGCG in the FLS was (0.08±0.02), which was declined significantly than the blank group (0.27±0.04) (P<0.05).Conclusion ① In vitro EGCG can inhibit T cell proliferation from peripheral blood and synovial fluid of RA patients and the TNF-α, IFN-γ, IL-1, IL-6 and IL-17A secretion are decreased. ② In vitro EGCG can inhibit the proliferation of RA FLS.
9.Preventing superficial bladder cancer from recurrence by intravesical instillation with Brucea Javanica oil emulsion after TUR-Bt operation
Feng TIAN ; He WANG ; Weijun QIN ; Kaichang ZHU ; Yue LIU
Chinese Journal of Rehabilitation Theory and Practice 2005;11(6):475-476
ObjectiveTo observe the effect of intravesical instillation of Brucea Javanica oil emulsion on recurrence of bladder cancer after transurethral resection of bladder tumor (TUR-Bt) operation.Methods187 patients with superficial bladder carcinoma after TUR-Bt operation were randomly divided into the group A (85 cases) and group B (102 cases). Patients of the group A were treated with instillation of Brucea Javanica oil emulsion; those of the group B were treated with mitomycin. A three-years following up was performed to observe the recurrence and side effects.ResultsAfter a 3-years following up, the recurrence rate of group A was 12.94%, lower than that of group B (34.31%). The side effects were seldom seen in the group A.ConclusionThe effect of intravesical instillation of Brucea Javanica oil emulsion to prevent the recurrence of bladder cancer after TUR-Bt operation is favorable.
10.The study on CASP 8-652 6N I/D polymorphism and susceptibility of 2 type diabetes mellitus
Xuemei HE ; Juan LI ; Xiaoping TANG ; Yongping HE ; Chuanfei HU ; Feng CHEN ; Qin WAN ; Zhuang CHEN
The Journal of Practical Medicine 2014;(19):3073-3075
Objective To investigate the association of cysteine aspartic acid specific protease 8 (CASP 8) gene-652 6N Insertion/Deletion polymorphisms and susceptibility of type 2 diabetes mellitus (T2DM). Methods CASP 8 gene -652 6N I/D polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing in 414 controls and 410 patients with T2DM. Results I/I, I/D and D/D genotype frequency were 56.5%, 38.9%, 4.6%in controls and 58.0%, 32.9%, 9.0%in T2DM group respectively (P<0.05). The risk in D/D genotype people was 1.916 times than I/I genotype (adjusted OR=1.916, 95%CI=1.199~3.054, P<0.05). The fasting blood sugar of D/D genotype people was significantly higher than that of I/D and I/I genotype people (P<0.05). Conclusions CASP 8 gene-652 6N I/D polymorphisms are associated with T2DM outbreak.