13C metabolic flux analysis(13CMFA)have been the research hotspots of metabolic engineering internationally due to its accuracy and applicability.It is vital that the measurement of 13C labelling pattern of proteinogenic amino acids for 13C metabolic flux analysis.To acquire 13C-labelling proteinogenic amino acids,Pseudomonas denitrifican which products Vitamin B12 was firstly fed with mimimal culture medium contained 20% U-13C and 80% natural glucose,after the culture reached a steady state,then about 20 mg biomass was hydrolyzed by 1 ml of 6 mol/L hydrochloric acid for 24h at 110℃.Then amino acids was separated,concentrated,evaporated in a vacuum,and derivatized with MBDSTFA,TBDMS-derivatized amino acids can be observed by GC-MS last we get 13C labelling pattern of fifteen aminio acids through mass spectrum.The experimental methods and sample preparation offers referential value for the development of 13C metabolic flux analysis in our courtry.

Objective To evaluate the efficacy of port catheter system(PCS)placement in pulmonary artery via percutaneous subclavicle vein treatment for multiple metastatic tumor in the two lungs and discuss the PCS technique.Methods Fifteen multiple metastatic tumor patients(13 hepatocellular carcinomas,one mandible grand adenocarcinoma,one oral bottom squamous carcinoma)were carried out with pulmonary artery PCS placement by way of percutaneous subclavicle vein.FPA/FPM/GP chemotherapy scheme were introduced every 4～6 weeks.Results The success rate of PCS placement technique was 93.3%(14/15).One case failed.Percutaneous subclavicle veins were performed 14 cases in right side and 1 in left one.Following up 2～43 months,2～7 chemotherapy cycles(mean 5 cycles)were accomplished,and the clinical CR and PR were achieved in 1 and 3 cases respectively with clinical efficacy rate 28.6%(4/14).Major side reaction was late wound healing in 1 case.Conclusion PCS placement in pulmonary artery treatment for multiple metastatic tumor in the two lungs is effective,and mastering operation technique is the key for increasing operation suc- cess rate.

A total of 105 patients with first-episode schizophrenia (male =51,female =54) and 99 normal controls (male =51,female =48) were included into this retrospective case-control study.Childhood trauma questionnaire-28 item short form (CTQ-SF) was used to assess the experience of childhood abuse.The result of binary logistic regression showed that emotional abuse (β =0.630,P ＜ 0.05) and emotional neglect(β =0.270,P ＜ 0.05) were included into the final model of predicting schizophrenia.It indicates that patients with first-episode schizophrenia experienced more early life stress than controls.Particularly emotional abuse and emotional neglect may play important roles in the onset of schizophrenia.

BACKGROUND: Silibinin has broad pharmaceutical effects, such as anti-free radicals, anti-lipid peroxidation, anti-lipoid oxidase, anti-glutathione (GSH) depletion, anti-neoplastic and serum lipid-lowering effects. Clinically, silibinin is often used in treating alcoholic liver disease. OBJECTIVE: To investigate the pharmacological mechanism of silibinin for alcoholic fatty liver in rats. DESIGN: Randomized and controlled study.SETTING: Guangzhou Hospital of Traditional Chinese Medicine.MATERIALS: The experiment was conducted at the Animal Experimental Laboratory of Guangdong Pharmaceutical Institute from August to October 2003. Totally 57 SD rats, without unusual bacteria, weighting (150±10)g and of either gender, were selected. Yiganling tablets containing 38.5 mg silibinin were produced by Zhuzhou No.3 Pharmaceutical Factory (Batch No. 20020808).METHODS: Among the 57 SD rats, 18 rats were regarded as normal control group. Rats in normal control group were administered with normal saline by gavage, and fed with normal food and distilled water in place of alcohol for 10 weeks. Rats in model group and silibinin group were fed with high-calorie food and 100 mL/L alcohol for 6 weeks to establish model of rat alcoholic fatty liver. The other rats were divided into model control group (n=18) and silibinin group (n=21). Rats in model control group were treated with distilled water while those in silibinin group were treated with 100 mg/kg silibinin. Meanwhile, 100 mL/L ethanol and hyperalimentation feed were given for 4 weeks. After animals were killed, TG, SOD, GSH and MDA levels were measured with liver suspension.MAIN OUTCOME MEASURES: Contents of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), total cholesterol (TC), triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), tumor necrosis factor (TNF)-α , and transforming growth factor (TGF)-β1.RESULTS: All the 57 rats entered the final analysis. Silibinin could inhibit the activities of serum AST, ALT and AKP [(2 550.5±400.1), (533.4±100.0), (2 217.1±750.2)nkat/L], and the differences were significant as compared with those in model control group [(3 600.7±666.8), (800.2±100.0), (2 900.6±1 333.6) nkat/L, P ＜ 0.05-0.01]. Contents of TG, LDL-C, TNF-α and TGF-β1 in silibinin group [(1.8±0.8), (0.17±0.04), (6.66±1.38), (24.1±4.1) mmol/L] were lower than those in model group [(2.8±1.4), (0.20±0.05), (7.81±1.06), (28.8±6.3) mmol/L] with significant differences (P ＜ 0.05-0.01). Silibinin could increase the content of HDL-C but decrease the contents of TG and MDA (P ＜ 0.05-0.01), and improve SOD activity as well as hepatocyte and fatty degeneration (P ＜ 0.01).However, it had no obvious effect on the content of reduced estathion (P ＞ 0.05).CONCLUSION: Silibinin can inhibitthe formation of alcoholic fatty liver in rats. The pharmacological mechanism of silibinin may involve anti-oxidation, removing free radicals, inhibiting lipid peroxidation, regulating blood lipid component, reducing fatty sediment in liver, and anti-immunoinflammation and anti-hyperplasia effects.

Objective To study the effects of inhibing AKT2 by siRNA on SMMC7721 liver cancer cells proliferation,apoptosis,migration and invasion.Methods The siRNA targeting AKT2 was designedandthe SMMC7721AKT2- siRNAplasmidwasconstructed andtransfected into SMMC7721 cells.The stable cell lines were screened by G418.The effects of AKT2 by siRNA on SMMC7721 liver cancer cells,growth inhibition was evaluated by MTT assay.Cell cycle was analyzed by flowcytometry (FCM).Protein of P27 and CyclinD1 was evaluated by Western-blot.The ability of migration and invasion was evaluated by wound healing and Transwell assay.ResultsThe growth of SMMC7721 cells was significantly inhibited by siRNA (P＜0.05).Flow cytometry display that AKT2 by siRNA can induce G1 phase arrest,the ratio of G1 phase increased homologously and S phase declined homologously.The protein of CyclinD1 was declined and the protein of P27 was increased by Western-blot.Wound healing and Transwell assay show that the ability of cells,migration and invasion was inhibited by AKT2 by siRNA.Conclusion AKT2 by siRNA can significantly inhibit the growth of SMMC7721 cells,arrest cell cycle.AKT2 by siRNA can inhibit the ability of invasion and migration of SMMC7721 cells.

Objective ToexploretheclinicalefficacyandsafetyofEmbospheremicrospherescombinedwithcoilsinbronchialarteryembolization (BAE)forpatientswithmassivehemoptysis.Methods FromJanuarytoDecember2017,63patientswith massivehemoptysiswere enrolledinourhospital,ofwhich29patientsreceivedEmbospheremicrospherescombinedwithcoilembolizationwereincludedasobservation group,andother34patientsreceivedembolizationwithEmbospheremicrospheresweretreatedascontrolgroup.Allthepatientswere followed-upin72hours.Results Theincidenceofhemostasisinobservationgroupwas96.55% within72hours,whichwassignificanthigher thanthatincontrolgroup (79.41% )(P<0.05).Themainreasonsforfailureweretherecanalizationofembolizedtargetvessel,the leakageofresponsiblevesselandpulmonaryarterypseudoaneurysm.Therewasnosignificantdifferenceoftheincidenceofadversereactions (mainlyincludingchesttightness,fever,chestpain)betweenthetwogroups(P=0.966).Conclusion Embospheremicrospherescombinedwith coilscansignificantlyimprovethehemostaticeffectofBAE,anddidnotincreasetheincidenceofpostoperativeadversereactions.

OBJECTIVETo investigate the molecular mechanisms underlying in the treatment of inflammatory bowel disease by Pulsatilla Decoction.

METHODSForty Wistar male rats were randomly divided into 5 groups( n = 8)control group, model group, model + positive control group (mesalazine), Pulsatilla Decoction treatment group, in addition, the Pulsatilla Decoction treatment group was divided into middle and high dose group. Intragastric administration was used in the positive control group and Pulsatilla Decoction treatment group. The expression of interleukin-1beta (IL-1beta), interleukin-6(IL-6) and tumor necrosis factor-alpha (TNF-alpha) were detected by real time PCR after extraction of RNA from colons.

RESULTSCompared with the model group, positive medicine and Pulsatilla Decoction group, especially high-dose group, could effectively inhibit the expression of IL-1beta, IL-6 and TNF-alpha.

CONCLUSIONPulsatilla Decoction could exert its effect in the treatment of inflammatory bowel disease by inhibiting the expression of proinflammatory cytokines.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Inflammatory Bowel Diseases ; drug therapy ; metabolism ; Interleukin-1beta ; genetics ; metabolism ; Interleukin-6 ; genetics ; metabolism ; Male ; Phytotherapy ; Pulsatilla ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

Aim To investigate the effect of TaorenHonghua drug pair on intervertebral disc degeneration (IVDD) in rats.Methods Fifty healthy Wistar rats were randomly divided into control group,model group,sham group,meloxicam group and Taoren-Honghua drug pair group,with 10 rats in each group.We established dynamic and static forces imbalance of cervical disc degeneration model or sham surgery in rats.12 weeks later,rats were intragastrically administered with meloxicam,Taoren-Honghua drug pair or saline for 30 days.C4/5 and C6/7 discs were harvested from rats.ABOG staining was used for observation of intervertebral disc morphology,real time PCR for mRNA expressions of type Ⅱ collagen (Col Ⅱ) and type Ⅹ collagen (Col Ⅹ),and immunohistochemical staining for Col Ⅱ and Col Ⅹ.Results Compared with model group,Col Ⅱ expression increased,while Col X expression decreased in chondrocyte of intervertebral disc in Taoren-Honghua-treated group(P ＜ 0.01).Conclusion Taoren-Honghua drug pair could delay the degeneration of cartilage endplate in rat intervertebral disc.

AIM:To investigate the effect of growth hormone receptor(GHR) knockdown on nuclear factor-κB (NF-κB) activity and inflammatory cytokine production stimulated by growth hormone (GH) in 3T3-L1 adipocytes. METHODS:The specific siRNA for GHR was transfected into 3T3-L1 adipocytes to silence GHR expressions. The effects of GH on NF-κB activation and inflammatory cytokine production in 3T3-L1 adipocytes transfected with siRNA-GHR or siRNA-control were measured by dual-luciferase system analysis,real-time RT-PCR and ELISA. RESULTS:The protein expression of GHR was diminished after transfection with GHR specific siRNA. Dual-luciferase reporter system analysis re-vealed that GHR knockdown resulted in attenuation of GH-stimulated NF-κB activation in the 3T3-L1 adipocytes. GHR knockdown ameliorated the GH-induced production of inflammatory cytokines TNF-α,IL-1β,IL-6,MCP-1 and MIP-1α in the 3T3-L1 adipocytes. CONCLUSION:Knockdown of GHR might be efficacious to prevent GH-induced inflammatory re-sponses in the 3T3-L1 adipocytes.

The influences of E23K polymorphism of inwardly rectifying K~+channel 6.2 (Kir6.2) gene on the clinical phenotype of type 2 diabetes and glucose-lowering effect of gliclazide were investigated.The result showed that E23K polymorphism did not influence glucose-lowering effect of gliclazide,but serum creatinine level of patients with K/K genotype was higher than those of E/E and E/K genotypes (P