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.

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 ; Chromatography, High Pressure Liquid ; Flavonoids ; chemistry ; Kaempferols ; Quercetin ; analogs & derivatives ; Rutin ; Tandem Mass Spectrometry ; Vitaceae ; chemistry

OBJECTIVETo assess the association between SIRT1 gene polymorphisms and the longevity phenomena in Yongfu region of Guangxi. In this case-control study, 500 individuals from Yongfu region of Guangxi were recruited. The subjects were divided into a longevity group (n=223, average age=93.17 U+00B1 3.08 yr) and a healthy control group (n=277, average age=46.92 U+00B1 17.12 yr). Polymerase chain reaction-high resolution melting curve (PCR-HRM) and DNA sequencing were used to determine the allelic and genotypic frequencies of rs3758391, rs3740051, rs2273773, rs4746720 and rs10997870 polymorphisms of SIRT1 gene in the two groups. The association between above polymorphisms and longevity was assessed.

RESULTSIn the longevity group, CT genotype of the rs4746720 locus was significantly more common than CC and TT genotypes (P=0.000, OR=2.098, 95%CI:1.412-4.117). However, no significant difference was found in the allelic and genotypic frequencies of rs3758391, rs3740051 and rs2273773 between the two groups.

CONCLUSIONThere is an association between rs4746720 of SIRT1 gene and longevity in Yongfu region of Guangxi.

Adult ; Aged ; Aged, 80 and over ; Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; China ; Female ; Gene Frequency ; Gene Order ; Genetic Association Studies ; Genotype ; Humans ; Longevity ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Sirtuin 1 ; genetics ; Young Adult

Animals ; Antimicrobial Cationic Peptides ; physiology ; Carrier Proteins ; physiology ; Cation Transport Proteins ; physiology ; Ceruloplasmin ; physiology ; Female ; Ferritins ; physiology ; Hemochromatosis Protein ; Hepcidins ; Histocompatibility Antigens Class I ; physiology ; Humans ; Iron ; metabolism ; Iron-Regulatory Proteins ; physiology ; Membrane Proteins ; physiology ; Placenta ; metabolism ; Pregnancy ; Transferrin ; physiology

Objective To assess the inlfuence of depth on liver stiffness measurement with real-time shear wave elastography (SWE) and determine the optimal depth for SWE in liver. Methods SWE of liver was performed on 89 healthy volunteers between May 2012 and November 2012. The depths of each liver were varied from 0 cm to 7 cm (from the liver capsule) in 1 cm increment and there were 8 depth groups in total. Then the elastic modulus of liver in each depth group were measured three times by SWE. The body mass index (BMI) and the distance from body surface to liver capsule were documented. The success rates and the mean elastic modulus of each group were calculated. Results The success rates of 0-7 cm were 0, 98.9%(88/89), 98.9%(88/89), 98.9%(88/89), 71.9%(64/89), 24.7%(22/89), 3.4%(3/89) and 0, respectively. The success rates were highest in 1 cm, 2 cm and 3 cm groups but signiifcant decreased with the increasement of depths in 4 cm, 5 cm and 6 cm groups ( 3 cm vs 4 cm, χ2=25.94, P<0.001; 4 cm vs 5 cm, χ2=39.68, P<0.001;5 cm vs 6 cm,χ2=16.79, P<0.001). The mean elastic modulus of 1 cm, 2 cm, 3 cm, 4 cm and 5 cm groups were (4.77±0.99), (4.68±0.99), (4.76±0.95), (5.19±1.10) and (5.41±0.95) kPa, respectively. The mean elastic modulus of 4 cm and 5 cm groups were signiifcant higher than those of 1 cm, 2 cm, 3 cm groups (4 cm vs 1 cm, t=-2.85, P=0.005;4 cm vs 2 cm, t=-3.49, P=0.001;4 cm vs 3 cm, t=-2.76, P=0.006;5 cm vs 1 cm, t=-3.13, P=0.002;5 cm vs 2 cm, t=-3.66, P=0.000;5 cm vs 3 cm, t=-3.05, P=0.003). In the group of 4 cm, the BMI and the distance from body surface to liver capsule of the volunteers performed successfully and unsuccessfully were (20.70±2.87), (22.07±2.42) kg/m2 and (1.45±0.25 ), (1.60±0.29) cm, respectively. In the group of 5 cm, the BMI and the distance from body surface to liver capsule of the volunteers performed successfully and unsuccessfully were (19.82±2.76), (21.49±2.72) kg/m2 and (1.35±0.21), (1.54±0.26) cm respectively. The BMI had no signiifcant difference between the successful and unsuccessful groups (t=-2.83, P=0.108 for 4 cm;t=0.77, P=0.709 for 5 cm), but the distance from body surface to liver capsule was signiifcantly different (t=26.51, P=0.012 for 4 cm;t=79.57, P=0.004 for 5 cm). Conclusions The success rates and elastic modulus were different at different depths. SWE should be performed at the depths of 1-3 cm from the liver capsule.

OBJECTIVE: To explore the correlation between the interleukin-17 (IL-17) level of peripheral blood and aggression of bipolar mania. METHODS: Thirty-six patients of bipolar mania were selected as experimental group by DSM-IV-TR and received treatment with quetiapine and lithium. Thirty-six healthy volunteers with similar age and gender were selected as control group. The level of IL-17 at baseline in each group and the level of IL-17 in the experimental group after treatment for 2, 4 and 8 weeks were detected by ELISA. RESULTS: The level of IL-17 in experimental group at baseline, after treatment for 2 and 4 weeks were all significantly higher than that in control group. After 8 weeks treatment, there was no significant difference between the two groups (P > 0.05). After 2, 4 and 8 weeks treatment, the total score and aggression score of Young Mania Rating Score (YMRS) were significantly lower than the baseline level (P < 0.05). In experimental group, the level of IL-17 was positively correlated with the two scores of YMRS at baseline (P < 0.05). CONCLUSION Bipolar mania may be related to the up-regulation of IL-17. The level of IL-17 is related to the severity of manic symptoms at baseline, especially aggression symptom.
Aggression/drug effects* ; Antipsychotic Agents/therapeutic use* ; Biomarkers/blood* ; Bipolar Disorder/drug therapy* ; Case-Control Studies ; Diagnostic and Statistical Manual of Mental Disorders ; Double-Blind Method ; Humans ; Interleukin-17/metabolism* ; Lithium Compounds/therapeutic use* ; Quetiapine Fumarate/therapeutic use* ; Treatment Outcome

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 ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Formates ; chemistry ; Tandem Mass Spectrometry ; methods ; Vitaceae ; chemistry ; Water ; chemistry

Objective: To analyze the features and patterns of acupoint selection in acupuncture-moxibustion treatment of chronic atrophic gastritis (CAG) by data mining technique. Methods:Relevant clinical studies published before 25 June, 2017 were searched in databases including PubMed, EMBASE, Cochrane library, Chinese Biomedical Literature Database (CBM), China National Knowledge infrastructure (CNKI), and Wanfang Academic Journal Full-text Database (Wanfang). Results:A total of 122 papers were included, involving 69 points. It was found that the top three points on the frequency list were Zusanli (ST 36), Zhongwan (CV 12) and Weishu (BL 21). The points selected were distributed in 11 meridians, in which the Stomach Meridian of Foot Yangming, Conception Vessel, and Bladder Meridian of Foot Taiyang ranked the top and accounted for 74.0％ of the total frequency. Of the involved specific points, Five Shu-Transmitting points, crossing points and Back-Shu points ranked the top, accounting for 47.1％. The analysis of association pattern has shown that Zusanli (ST 36) and Zhongwan (CV 12) won the highest support rate in the paired groups; Zusanli (ST 36), Weishu (BL 21) and Zhongwan (CV 12) had the highest support rate among the point groups. The Five Shu-Transmitting points and the Lower He-Sea points had the highest support rate among the specific point groups. Conclusion: The data mining results of the studies on acupuncture-moxibustion for CAG are substantially in line with the acupuncture-moxibustion treatment theories in traditional Chinese medicine. The results can reflect the acupoint selection patterns in treatment of CAG and provide reference for acupuncture-moxibustion treatment of CAG in clinic.

OBJECTIVETo investigate the cell viabilities of vascular smooth muscle cells and vascular endothelial cells stimulated by cigarette smoke extract(CSE) .

METHODSThe CSE was prepared by smoke-bubbled phosphate buffered saline(PBS) generation.After culturing cells with different concentrations of CSE, we used the cell counting kit-8 to determine the cell viability.The expression levels of c-jun and cyclinD1 were analyzed through Western blot.The c-jun plasmid was transfected to detect the change of cyclinD1 expression.

RESULTSThe smooth muscle cell viability increased when the CSE concentration ranged 0.625%-10%, whereas the endothelial cells viability decreased when exposed to the CSE concentration. After exposure to CSE for 48 hours, there was no difference in c-jun expression between toxin group and PBS group;however, the expression of p-c-jun in the smooth muscle cells significantly increased in the toxin groups than in the PBS group(P<0.05) and the expression of p-c-jun in the vascular endothelial cells significantly decreased(P<0.05) . The level of cyclinD1 significantly increased after exposed to CSE, and its expression level also increased in respond to the c-jun overexpression.

CONCLUSIONCSE can enhance the proliferation of vascular smooth muscle cells and decrease in the activity of endothelial cells proliferation, which may be explained by the phosphorylation of c-jun and the expression of cyclinD1.

Cell Proliferation ; drug effects ; Cell Survival ; Cells, Cultured ; Cyclin D1 ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Humans ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Proto-Oncogene Proteins c-jun ; metabolism ; Tobacco ; adverse effects

BACKGROUND: Proximal femoral nail anti-rotation is widely used to treat various intertrochanteric fractures. Although its operation trauma is small, and the blood loss of perioperative period is still large. Tranexamic acid has been gradually used to reduce the bleeding of intertrochanteric fracture. The effectiveness and safety of reducing blood loss during perioperative period were not reported. OBJECTIVE: To explore the safety and efficacy of tranexamic acid on perioperative blood loss in patients with intertrochanteric fracture undergoing proximal femoral nail anti-rotation. METHODS: One hundred and eight patients with intertrochanteric fracture undergoing proximal femoral nail anti-rotation were selected from First Affiliated Hospital, Guangzhou University of Chinese Medicine between January 2015 and January 2017. Among all the subjects, 52 patients who received the operation before January 2016 served as the control group and 56 patients who received the operation after January 2016 were selected as the treatment group. Half an hour before operation, patients in the treatment group received 1 g tranexamic acid dissolved in 250 mL normal saline by intravenous dropping; patients in the control group just received 250 mL normal saline by intravenous dropping. The bleeding volume, blood transfusion volume, hemoglobin, hematocrit, coagulation index, D-dimer levels and complications were compared between the two groups. RESULTS AND CONCLUSION: (1) During perioperative period, actual blood loss, intraoperative blood loss, dominant blood loss, recessive blood loss, volume of drainage, blood transfusion volume and blood transfusion rate were lower in the treatment group than in the control group (P < 0.05). (2) There was no statistically significant difference in the hemoglobin and hematocrit between the two groups before operation (P > 0.05). The hemoglobin and hematocrit of the two groups gradually decreased after the operation, and there was a slight improvement in the fifth day after surgery. At postoperative 2 hours, 1, 3 and 5 days, the hemoglobin and hematocrit of the treatment group were higher than in the control group (P < 0.05). At preoperation and each time point postoperation, prothrombin time, activated partial thromboplastin time, and fibrinogen levels were not statistically significant between the two groups (P > 0.05). Postoperative D-dimer levels in the two groups were significantly higher than preoperation, and there was a return on the fifth day. There was no statistically significant difference between groups at preoperation and each time point of postoperation (P > 0.05). (3) The results suggest that the tranexamic acid can effectively reduce the dominant and recessive blood loss in patients with the intertrochanteric fracture, and it is safe and effective.