To establish a UPLC-MS/MS method for the simultaneous determination of geniposide, genipin 1-O-beta-D-gentiobioside and geniposidic acid in rat brains and study the brain pharmacokinetics of the three iridoid glycosides in stroke rat after the oral administration of Xingnaojing. In this experiment, brain samples were precipitated with protein for twice. Acquity BEH C18 column was adopted, with acetonitrile-0.1% formic acid-water as the mobile phase for gradient elution. ESI source was adopted for mass spectra; multiple reaction monitoring (MRM) was conducted to detect negative ions. The time for sample analysis was 3.5 min. the results showed good linear relations among the three iridoid glycosides, with the extraction recovery between 99.6% and 114.3%, good intra- and inter-day precisions and accuracies and stability in line with the requirements. The t1/2 and MRT in the three components were similar in brains of stroke rats. Geniposide and genipin 1-O-beta-D-gentiobioside showed double peaks; where as geniposidic acid showed a single peak. In conclusion, the method is so specific, sensitive, accurate and reliable that it can be used to study the brain pharmacokinetics of Xingnaojing oral preparation.
Animals ; Brain ; metabolism ; Brain Chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Iridoids ; chemistry ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Candida shehatae xyl1 gene and Pichia stipitis xyl2 gene were amplified by PCR and the xyl1 and xyl2 were both placed under the promoter GAL of vector pYES2 to produce the recombinant expression vector pYES2-P12. Subsequently the pYES2-P12 vector was transformed into S. cerevisiae YS58 by LiAc to produce the recombinant yeast YSS8-12. It was indicate that the recombinant yeast YSS8-12 could converse xylose to ethanol with the xylose consumption rate of 81. 3%.

Aim To investigate the effect of dihydromyricetin on canine carotid artery and the underlying mechanism.Methods The in vitro isometric tension measurement technique was employed to investigate the effect of dihydromyricetin on canine carotid artery rings.Laser scanning confocal microscope technique was used to measure the dynamic change of intracellular calcium concentration in single VSMC.Results Dihydromyricetin(1～300 ?mol?L-1)caused a concentration-dependent contraction of both endothelium-intact and endothelium-denuded rings.This constrictive effect was attenuated in Ca2+-free solution(P

OBJECTIVETo examine the prevalence changes of diabetes mellitus (DM) and impaired fasting glucose (IFG) from 2003 to 2010 in the health check-up subjects in Shanghai.

METHODSHealth check-up subjects were divided into ten groups by sex and each 5 years old, and the prevalence of crude DM, crude IFG were calculated first. According to Chinese sex and age structure of China Population Statistics Yearbook 2006, sex and age standardized DM and standardized IFG were computed.

RESULTSIn the same year, the prevalences of crude DM and IFG increased with increasing age for both male and female, reached the summit at 60 - 69 age group, when at ≥ 70 age group, they had a down trend and were still at higher level. The prevalences of crude DM were 3.99% (986/24 699) in male and 1.61% (176/10 948) in female in 2003, and were 7.85% (3366/42 899) and 2.55% (531/20 820) in 2010. The prevalences of crude IFG were 9.97% (2462/24 699) in male and 5.88% (644/10 948) in female in 2003, and were 30.96% (13 283/42 899) and 17.16% (3573/20 820) in 2010. The prevalences of age standardized DM in 2003 and 2010 were 3.89% and 6.90% for male (χ(2) = 371.89, P < 0.01), 2.12% and 3.23% for female (χ(2) = 29.32, P < 0.01), respectively. The prevalences of age standardized IFG in 2003 and 2010 were 9.51% and 28.55% (χ(2) = 3865.56, P < 0.01) for male, 6.97% and 17.88% (χ(2) = 790.81, P < 0.01) for female. The prevalences of age and sex standardized DM were 3.00% and 5.05% (χ(2) = 385.39, P < 0.01), and prevalences of age and sex standardized IFG were 8.23% and 23.17% (χ(2) = 4480.21, P < 0.01).

CONCLUSIONFrom 2003 to 2010, prevalences of DM and IFG had increased greatly. It concluded that first-level prevention of DM for health check-up subjects should start from youth, and should lay emphasis on population of IFG, especially for male.

Adult ; Aged ; Aged, 80 and over ; Blood Glucose ; analysis ; China ; epidemiology ; Diabetes Mellitus ; epidemiology ; Fasting ; Female ; Glucose Tolerance Test ; Halfway Houses ; Humans ; Male ; Middle Aged ; Physical Examination ; Prediabetic State ; epidemiology ; Prevalence

OBJECTIVETo evaluate the effect of alpha-galactosidase A (Gla) deficiency on FV Leiden (FVL) associated thrombosis in vivo.

METHODSTo generate the mice carrying mutations in Gla and FVL and analyze the tissue fibrin deposition in organs and thrombosis.

RESULTSIn the presence of FVL, Gla deficiency greatly increased tissue fibrin deposition compared with that in wild-type [Gla(-/0) FV(Q/Q) vs. Gla(+/0) FV(Q/Q) = (0.24 +/- 0.07)% vs. (0.086 +/- 0.049)%, P < 0.0001; Gla(-/-) FV(Q/Q) vs. Gla(+/+) FV(Q/Q) = (0.32 +/- 0.03)% vs. (0.06 +/- 0.005)%, P < 0.05]. With Gla deficiency, the number of thrombi on organ sections in FVL mice was significantly increased [(Gla(-/-) FV(Q/Q) and Gla(-/0) FV(Q/Q)) vs. (Gla(+/+) FV(Q/Q) and Gla(+/0) FV(Q/Q)) = 1.9 +/- 0.7 vs. 0.3 +/- 0.1, P < 0.05].

CONCLUSIONSGla deficiency could be an important genetic modifier for the enhanced thrombosis associated with FVL.

Animals ; Fabry Disease ; genetics ; Factor V ; genetics ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Mutation ; Thrombosis ; genetics ; pathology

Objective To investigate the diagnostic value of prenatal ultrasonography and magnatic resonance imaging in the fetal intracranial hemorrhage. Methods The 20 antenatal diagnosed ICH cases was collected from 31 200 prenatal diagnosis units in Guangzhou Women and Children′s Medical Center from July 2012 to June 2014. Maternal characteristics, ultrasound, and magnetic resonance imaging findings, clinical course, and postnatal outcome were reviewed. Results Twenty consecutive cases of fetal ICH were evaluated. All cases were diagnosed at mid or third trimester. Transabdominal ultrasound showed 9 cases of hyper echoic lesions in the lateral ventricle, 2 cases of hypoechonic lesions, 2 cases of irregular mixed echo in the parenchyma, 1 case of hyperechoic cerebellar hemisphere with infarction, and 1 case of abnormal choroid plexus. Seventeen cases were associated with ventriculomegaly, brain compression or brain midline displacement. MRI showed the nodular, patchy or linear low signal on T2WI and high signal on T1WI. Some cases were combined with cortex lesions or abnormal parenchyma. ICH was complicated with other structural abnormalities:1 case of cleft palate, 2 cases of spinal deformity and 3 cases of other cerebral cortical malformations. Prenatal diagnosis results were:2 cases diagnosed as positive cytomegalovirus infection and no chromosome abnormalities found in all cases. The follow up results were:1 case was lost, 16 cases were terminated after prenatal diagnosis. Among the 3 survival cases, 1 case has the neurological complication and the other two were normal till now. Conclusions Fetal intracranial hemorrhage has some image features on ultrasound and magnetic resonance. Ultrasound showed hyper echoic lesions with ventriculomegaly. MRI showed the nodular, patchy or linear low signal on T2WI and high signal on T1WI. MRI may contribute to the accuracy of diagnosis, particularly in bleeding site. The regular ultrasonic monitoring is helpful to improve the detection rate.

Aim To investigate the effects of dihydromyricetin on the apoptosis in AGZY-83-a tumor cells and its relevant mechanisms.Methods The survival rate of AGZY-83-a cells was assayed by MTT dye reduction.The cellular apoptosis was analyzed by TUNEL method and caspase-3 activity.The intracellular free calcium i was detected to explore the apoptotic mechanism of dihydromyricetin.Results In the MTT assay,dihydromyricetin 50?mol?L-1 significantly inhibited survival ratios of AGZY-83-a cells at dose-and time-dependent manner.It was demonstrated in TUNEL assay that dihydromyricetin could induce the apoptosis of AGZY-83-a cells in a concentration-dependent manner.The examination of Caspase-3 activity indicated that the dihydromyricetin could induce the apoptosis of AGZY-83-a cells,which was dose-dependent activation of Caspase-3 in AGZY-83-a cells.The detection of the intracellular i showed that the average FI of the i could be markedly increased to 20-fold as the basic condition.Conclusions Dihydromyricetin can induce the apoptosis in AGZY-83-a cells,which is associated with the overload of the intracellular i.

OBJECTIVETo evaluate the efficacy and safety of trastuzumab in combination with chemotherapy versus chemotherapy alone in the first-line treatment of HER-2-positive advanced gastric or gastro-oesophageal junction cancer.

METHODSFifteen Chinese research centers are involved in the BO18255 (ToGA) study. Patients with gastric or gastro-oesophageal junction cancer were eligible for inclusion if their tumor showed overexpression of HER-2 protein by immunohistochemistry +++ or FISH-positive. Patients were randomly assigned in a 1:1 ratio to receive a chemotherapy regimen consisting of capecitabine or 5-FU plus cisplatin or chemotherapy in combination with intravenous trastuzumab. The primary endpoint was overall survival.

RESULTSEighty-five Chinese patients were enrolled in this study, of whom 84 were included in the primary analysis: trastuzumab plus chemotherapy (FP/H) (n = 36) and chemotherapy alone (FP)(n = 48). The median follow-up was 15.2 months in the FP/H group and 14.2 months in the FP group. The median survival time was 12.6 months in the FP/H group compared with 9.7 months in the FP group [hazard ratio 0.72, 95%CI (0.40; 1.29)]. Grade 3/4 adverse events were higher in the FP/H(63.9%)than FP (47.9%) groups, including neutropenia, vomiting and nausea. Two mild cardiac adverse events occurred in the FP/H group. Severe adverse events occurred in 3 cases of both two groups, respectively.

CONCLUSIONSAddition of trastuzumab to chemotherapy is well tolerated and shows improved survival in Chinese patients with advanced gastric or gastro-oesophageal junction cancer. These results are consistent with the results of ToGA whole population trial. Trastuzumab in combination with chemotherapy can be considered as a new option for patients with HER-2-positive advanced gastric or gastro-oesophageal junction cancer.

Aged ; Antibodies, Monoclonal, Humanized ; administration & dosage ; adverse effects ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Capecitabine ; China ; Cisplatin ; administration & dosage ; adverse effects ; Deoxycytidine ; administration & dosage ; adverse effects ; analogs & derivatives ; Esophageal Neoplasms ; drug therapy ; pathology ; Esophagogastric Junction ; Female ; Fluorouracil ; administration & dosage ; adverse effects ; analogs & derivatives ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Nausea ; chemically induced ; Neoplasm Staging ; Neutropenia ; chemically induced ; Receptor, ErbB-2 ; metabolism ; Remission Induction ; Retrospective Studies ; Stomach Neoplasms ; drug therapy ; pathology ; Survival Rate ; Trastuzumab ; Vomiting ; chemically induced

OBJECTIVETo evaluate the role of leptin in neointimal formation and related mechanisms.

METHODSFemoral arterial injury was induced in wild-type (Wt, n = 10), leptin-deficient (Lep(-)/-, n = 12), and leptin receptor-deficient (LepR(-)/-, n = 10) mice. Leptin treatment studies (tail vein injection of adenovirus expressing murine leptin on the RSV promoter, ad-leptin) were performed on Lep(-)/- (n = 5) and LepR(-)/- (n = 4) mice. Intimal (I) and medial (M) areas were measured and the ratio of I/M was calculated. Smooth muscle cells were detected by smooth muscle alpha-actin staining using an alpha-actin monoclonal antibody. Cellular proliferation was analyzed with BrdU Staining Kit and the number of BrdU-positive cells was counted manually. Plasma leptin level was measured by ELISA.

RESULTSThe I/M ratio of Lep(-)/- and LepR(-)/- mice was significantly lower than that in Wt separately (Lep(-)/- vs. Wt = 0.80 +/- 0.14 vs. 1.50 +/- 0.22, P < 0.01; LepR(-)/- vs. Wt = 0.55 +/- 0.20 vs. 1.50 +/- 0.22, P < 0.05). Plasma leptin level was significantly increased in Lep(-)/- and LepR(-)/- mice post leptin treatment. I/M was significantly increased in Lep(-)/- mice receiving ad-leptin compared with untreated Lep(-)/- mice (P < 0.05), while I/M was similar between LepR(-)/- mice with and without ad-leptin treatment (P > 0.05). The changes on number of positive alpha-actin and BrdU stained smooth muscle cells were consistent with the neointimal formation findings in various groups.

CONCLUSIONSMice lacking leptin or the leptin receptor were protected from neointimal formation following vascular injury. Leptin treatment increased neointimal formation in Lep(-)/- but not in LepR(-)/- mice, suggesting leptin receptor activation and vascular smooth muscle cell proliferation played a pivotal role on neointimal formation post-injury in this model, giving an evidence that high plasma leptin level is a risk factor for neointimal formation.

Actins ; analysis ; Animals ; Cell Proliferation ; Leptin ; blood ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Muscle, Smooth, Vascular ; pathology ; Receptors, Leptin ; metabolism ; Tunica Intima ; pathology

OBJECTIVEIn order to investigate the role of shear stress in the regulation of endothelial function, we assessed here effects of shear stress on tissue-type plasminogen activator in human endothelial progenitor cells (EPCs).

METHODSThe peripheral blood mononuclear cells were separated from healthy adult and inducted into EPCs, which were identified by double staining for the fluorescent labeled acetylated-LDL and lectin. EPCs were seeded on the small diameter artificial vessels, and then divided into four different experimental groups including stationary group, low-flow shear stress group (5 dyn/cm(2)), medium-flow shear stress group (15 dyn/cm(2)) and high-flow shear stress group (25 dyn/cm(2)). The levels of t-PA in EPC culture medium at 0 hour, 5 hours, 10 hours, 15 hours, 20 hours and 25 hours after culture were measured by enzyme-linked immunosorbent assay.

RESULTSThe peripheral blood mononuclear cells differentiated into EPCs after induction, which were positively labeled by fluorescent acetylated-LDL and lectin. Shear stress enhanced production of the t-PA by EPCs, which was paralleled to levels and times of shear stress.

CONCLUSIONSShear stress increases t-PA secretion by human EPCs, suggesting that shear stress not only regulates vascular endothelial function but also participates in the pathogenesis of arteriosclerosis.

Adult ; Endothelial Cells ; secretion ; Humans ; Stem Cells ; secretion ; Stress, Mechanical ; Tissue Plasminogen Activator ; metabolism