A comprehensive HPLC-DAD-MS method was developed to study the chemical components of semi-bionic extract of Coptis-Evodia herb couple. The extract was isolated on a Hypersil BDS C18 column (4.6 mm x 200 mm, 5 microm) using acetonitrile-ammonium formic buffer as mobile phase by gradient elution. Detection was performed on DAD and MS equipped with an electrospray ionization (ESI) source by full scan and product full scan on positive mode. The chromatogram of Coptis-Evodia showed seventeen main peaks, eight of which were from Evodia while the others were from Coptis. By comparison of the retention time, the on-line UV spectra and MS spectra, four peaks were identified as jatrorrhizine, hydroxevodiamine, palmatine and berberine, and three peaks were deduced as epiberberine, columbamine and coptisine. In addition, berberine and palmatine were quantitatively determined. No new component was created in the semi-bionic extract of the herb couple, yet the solubilities of berberine and palmatine decreased.
Berberine ; analogs & derivatives ; chemistry ; isolation & purification ; Berberine Alkaloids ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal ; Evodia ; chemistry ; Plant Extracts ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Objective To know the influencing factors for the severity of motor vehicle road traffic injures ,and to provide scientific basis for making appropriate intervention measures.Methods 7 977 patients with motor vehicle road traffic injuries who were first diagnosed in injury surveillance sentinel hospital of Tongxiang City in 2012 and 2013 were selected, and the factors influencing the severity of injury were analyzed.Results The multivariate logistic regression analysis showed an increased risk of the injure severity among the people aged over 65 years old,alcohol drinking before the injury and migrant workers,the values of OR(95%CI)were 3. 231(2. 091 -4. 993),2. 796(1. 697-4. 607)and 1. 424(1. 217-1. 666)respectively.However,women,motor vehicle drivers,collision with non motor vehicle driver and using safety belt may contribute to the less serious damage,and the values of OR (95%CI)were 0. 837 (0. 753 -0. 930 ),0. 769 (0. 622-0. 952),0. 753 (0. 598-0. 948)and 0. 691 (0. 492-0. 970)respectively.Conclusion The risk factors for the severity of motor vehicle road traffic injures are elderly people,migrant workers and alcohol drinking.The safety education should be provided among the people and the targeted interventions should be developed and implemented.

Objective To explore the clinical value of 99Tcm-MIBI G-MPI in patients diagnosed with myocardial bridging(MB) by CTA. Methods Forty-five patients with MB and 17 normal controls diagnosed by CTA(64 slices CT) were included. All patients underwent rest 99Tcm-MIBI G-MPI and 17 MB patients and 9 normal controls also underwent stress 99Tcm-MIBI G-MPI. Myocardial ischemia, function and wall motion were assessed. G-MPI results were compared with CTA results by χ2 test, Fisher exact test and t test. Results In patients with MB, the positive rate of abnormal perfusion by gated stress 99Tcm-MIBI G-MPI was 64.7% (11/17) and 41.2% (7/17) using quantitative analysis and visual evaluation respectively; while the data were 42.2% (19/45) and 22.2% (10/45) by rest G-MPI (P=0.035). The positive rate by rest G-MPI in MB patients was significant different among mural coronary arteries of different depths and different locations. By quantitative analysis of the stress G-MPI, the reversible, fixed, and mixed ischemia patients were 4 (35.3%), 6 (23.5%) and 1 (5.9%) respectively; the reversed, reversible and fixed abnormity of wall motion was found in 4 (23.5%), 4 (23.5%) and 2 (11.8%) patients respectively; the reversed, reversible and fixed wall thickening were found in 6 (35.3%), 5 (29.4%) and 1 (5.9%) patients respectively. There was no significant difference in left ventricular ejection fraction and peak filling rate between MB patients and normal controls in both rest and stress studies (t: from -0.564 to 1.292, all P>0.05). Conclusion The G-MPI may be useful for the evaluation of myocardial ischemia and myocardial function simultaneously in patients with MB.

Circular RNA(circRNA)is a type of noncoding RNA with tissue specificity and high stabil-ity, which forms a closed continuous loop and is abundantly expressed in tissue cells. According to re-cent research, the regulatory function of circRNA elucidating in the occurrence and development of dis-ease shows a potential for diagnosing clinical disease and revealing disease mechanism. This paper re-views the biological characteristics, analysis methods of circRNA and its research progress in clinical ap-plication as biomarker, and outlooks its application in the field of forensic medicine.

Objective To exam the influence of psychological factors,occupational factors and dietary habits on malignant tumors.Methods A total of 2 471 patients with malignant tumor were selected from 2010 to 2012 and 1 to 1 matching method was used controlling sex and age distribution in the case and control groups.Data collected by interviewing was analyzed by univariate and multivariate conditional logistic regression.Results 12 related factors including 6 psychological factors were found in relation to malignant tumor after univariate logistic regression analysis.The multivariate logistic regression analysis showed that harmful occupational history (OR =1.731,95%CI:1.337 ~2.242),often having fried food (OR =1.889,95%CI:1.398 ~2.551),heavy salty diet(OR =1.457,95%CI:1.169 ~1.818),psychological problems (OR =1.274,95%CI:1.003 ~1.617),poor marital life (OR =2.007,95%CI:1.111 ~3.623)and lack of self -regulation (OR =1.460,95%CI:1.189 ~1.793 )were the risk factors.Conclusion Malignant tumor was related to a variety of exposure factors and psychological factor has become an important risk factor for malignant tumor in Tongxiang city.

Objective To know the plasma lipids level and influencing factors among adults in Tongxiang city,and to provide evidence for prevention and intervention of dyslipidemia.Methods Multistage stratified cluster sampling method was conducted to select participants in Tongxiang city.Questionnaire interview,physical examination and blood lipid detection were performed among the selected subjects.Models of the univariate and multivariate unconditional logistic regressions were used to analyze the influencing factors of dyslipidemia.Results A total of 600 people were involved in this study.The prevalence and standardized rates of dyslipidemia were 36.33% and 36.11 %,and abnormal rates of HDL -C,TG,TC and LDL -C were 27.33%,15.67%,4.83% and 4.83%,which standardized were 29.17%,15.1 3%,3.1 8% and 3.12%, respectively.The abnormal rate of LDL -C in females was higher than that in males (P <0.05).Borderline abnormal rate of TC and dyslipidemia rates were increased with age (P <0.05),and the abnormal rate of HDL -C were decreased with age (P <0.05).Multivariate logistic regression analysis showed that BMI (OR =3.398),diabetes mellitus(OR =1.694), hypertension (OR =1.717)and total static time(OR =1.712)were the risk factors for dyslipidemia.Conclusion The prevalence of dyslipidemia was high among the adults in Tongxiang city,especially the abnormity of HDL -C and TG. Community comprehensive prevention and control of dyslipidemia should be carried out according to its risk factors.

BACKGROUNDWe determined the diagnosis of hereditary hemorrhagic telangiectasis (HHT) in a suspected HHT family, identified ALK1 gene mutation and established a gene diagnosis method of HHT. The level of related plasma proteins (transforming growth factor beta and thrombomodulin) were also analyzed.

METHODSBleeding history and family history were collected; Dilatant nasal mucosal capillaries in proband were observed under nasal cavity endoscope; exons 3, 7, 8 of ALK1 gene in proband and her family members were amplified with polymerase chain reaction (PCR), and the PCR products were analyzed. Using enzyme-linked immunosorbent assay (ELISA), plasma TGF-beta1 and TGF-beta2 concentrations were measured. Plasma thrombomodulin (TM) level was detected by Western blotting.

RESULTSOf all family members, four had epstaxis, two had evident telangiectases on skin or mucosa. Gene screening results showed that C to T substitution at position 1231 in exon 8 of ALK1 gene (CGG-->TGG) existed in proband, her affected brother and their father. The mutation did not exist in proband's sister-in-law and nephew. Plasma TGF-beta1 concentrations in the affected HHT was 20,538, 17,194, 13,131 pg/ml, while that of normal control and unaffected family members was 15,950, 20,297, 12,836 pg/ml, respectively. Plasma TGF-beta2 in HHT patients was 14,502, 9550, 10,592 and that of normal controls 8579, 20,297, 7680 pg/ml respectively. Level of plasma TM was in HHT subjects significantly lower than in normal subjects.

CONCLUSIONSChinese HHT individuals have mutant ALK1 gene, a C1231T variation on exon 8 of ALK1 is responsible for HHT clinical phenotypes in this family. ALK1 gene analysis, together with special clinical phenotypes and family history, provides a reliable method in diagnosing HHT. In affected HHT subjects, plasma TGFbeta levels were not obviously different from those of normal subject; while plasma TM concentration was significantly lower than that in normal subjects. The significance and mechanism remain to be elucidated.

Activin Receptors, Type I ; genetics ; Activin Receptors, Type II ; Aged ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Telangiectasia, Hereditary Hemorrhagic ; blood ; genetics ; Thrombomodulin ; blood ; Transforming Growth Factor beta ; blood

OBJECTIVETo explore the biological characteristic of third-party-derived tolerogenic DC(tDC) and the influence of third-party-derived tDC on acute graft-versus-host-disease (aGVHD) following allogeneic bone marrow transplantation (allo-BMT) in mice.

METHODStDC from bone marrow cells of D1 mice was cultured with low doses of GM-CSF, IL-10 and TGF-β1D1. The phenotype, expression of cytokines and function associated molecules were identified with FACS and RT-PCR. Mixed lymphocyte reaction was applied to analyze the influence of third-party-derived tDC on allo-CD4(+)T cells proliferation in vitro. Different doses of D1-tDC were adoptive transferred in the aGVHD model in allogeneic BMT which B6 mice as donors and D2 mice as recipients. Survival time, clinical GVHD score and the levels of Th1/2 cytokines in serum were monitored after allo-BMT using the aGVHD model as control.

RESULTStDC expressed lower levels of MHC II and co-stimulatory molecules, such as CD80, CD86 and CD40, even when stimulated by LPS. The results by RT-PCR indicated that tDC expressed low levels of IL-12p40 and high levels of immunosuppressive molecules, such as IL-10, TGF-β, Fas Ligand, indoleamine 2, 3-dioxygenase (IDO) and arginase. In the allogeneic MLR, third-party tDC suppressed allo-CD4(+)T cells proliferation, which was relative to the dose of tDC. In the B6→D2 mouse model, all aGVHD mice died within 18 days. Remarkably, if 10(4) third-party tDC were transferred, 60% mice survived at least 60 days. When the doses of tDC were reduced to 10(3) cells, only 20% of mice survived day 60, and when increased tDC to 10(5), all of the mice died within day 37 after allo-BMT. The cytokine levels in serum indicated that 10(4) tDC-treated mice secreted in vivo high level of IL-10 21d after BMT (P < 0.05), the levels of IL-10 in 10(3), 10(4) and 10(5) tDC-treated mice were (114.23 ± 7.78), (646.18 ± 212.02), (121.97 ± 10.47) ng/L, respectively.

CONCLUSIONThird-party tDC could suppress allo-CD4(+)T cells proliferation in vitro and prevent aGVHD in allogeneic BMT mode, which may be mediated by modulating tolerogenic cytokines secretion, such as IL-10. And this effect was associated with the dose of tDC. Adoptive therapy by transfusing third-party tDC cultured with low doses of GM-CSF, IL-10 and TGF-β1 could significantly prolong the survival of recipients and prevent aGVHD in allogeneic BMT.

Animals ; Bone Marrow Transplantation ; adverse effects ; CD4-Positive T-Lymphocytes ; cytology ; Cell Proliferation ; Dendritic Cells ; cytology ; immunology ; metabolism ; Graft vs Host Disease ; prevention & control ; Interleukin-10 ; immunology ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Transforming Growth Factor beta1 ; immunology ; Transplantation, Homologous

AIM:To investigate the effect of urantide on the liver function and histomorphology in the rats with atherosclerosis (AS).METHODS:The AS Wistar rat model was induced by intraperitoneal injection of vitamin D3 (VD3) and feeding with high-fat diet.The rats were randomly divided into normal control group, AS model group, positive medicine group and urantide group.The liver function indexes of the rats were measured by biochemical test, and the pathological changes of the aorta and liver of the rats were observed by hematoxylin-eosin (HE) staining.The mRNA expression of urotensinⅡ (UII) and GPR14 at mRNA and protein levels in rat livers was determined by RT-qPCR and Western blot.RESULTS:The levels of alanine aminotransferase (ALT) , aspartate aminotransferase (AST) , γ-glutamyltransferase (γ-GT) , lactate dehydrogenase (LDH) , total bilirubin (TBIL) , indirect bilirubin (IBIL) and alkaline phosphatase (ALP) in AS model group were significantly increased compared with normal control group (P<0.05).The above indexes in urantide group were remarkably decreased compared with AS model group (P<0.05).No change of the levels of direct bilirubin (DBIL) , total protein (TP) , globulin (GLB) and albumin (ALB) in each group was observed.Urantide postponed hepatocyte fatty degeneration and repaired hepatocyte injury in the AS rats.Compared with normal control group, the mRNA and protein levels of UII and GPR14 in the liver were significantly increased in AS model group (P<0.05).With the prolongation of dosing time, the mRNA and protein levels of UII and GPR14 in the liver were significantly decreased in urantide group compared with AS model group (P<0.05).CONCLUSION:Urantide significantly attenuates the liver damage caused by liver fatty degeneration in AS rats.