This study was designed to investigate inhibitory effects and possible mechanisms of snake venom tripeptide (pENW) on platelet adhesion in order to promote the development of a novel anti-platelet therapy. To study the inhibitory effects of pENW on platelet adhesion, washed platelets pre-incubated with pENW (116.5-466.2 μmol x L(-1)) were used to test the ability of platelet adhesion to fibrinogen. Effect of pENW on fibrin clot retraction was also tested. Effect of pENW on platelets viability was tested by MTT assay. Effect of pENW on reactive-oxygen species (ROS) levels of platelet was studied by flow cytometry assay. Calcium mobilization in Fura-2/AM-loaded platelets was monitored with a spectrofluorimeter. Cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP), thromboxane A2 (determined as its metabolite thromboxane B2) were measured using enzyme immunoassay kits. Akt, ERK and p38 phosphorylation were tested by Western blot. The results showed that pENW inhibited platelet adhesion and fibrin clot retraction in a concentration-dependent manner without cytotoxicity. Intracellular cGMP and cAMP in both resting and thrombin-activated platelets were increased by pENW. In addition, pENW attenuated intracellular Ca2+ mobilization and TXA2 production in platelets stimulated by thrombin. As shown by Western blot assay, Akt, ERK and p38 phosphorylation in thrombin-induced platelet were attenuated by pENW. However, inhibitory effects of pENW had nothing to do with ROS. Thus, pENW exhibited a significant inhibition on platelet adhesion to fibrinogen, which means pENW could block the first step of thrombosis as while as retard the more stable clot formation. The mechanisms of pENW on inhibition platelet adhesion might be related to instant regulations, such as protein kinases.
Blood Platelets ; drug effects ; Blotting, Western ; Calcium ; metabolism ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Flow Cytometry ; Phosphorylation ; Platelet Aggregation ; drug effects ; Reactive Oxygen Species ; metabolism ; Snake Venoms ; chemistry ; Thromboxane A2 ; metabolism ; Thromboxane B2 ; metabolism

?AIM: To study the association of the single nucleotide polymorphism ( SNP) rs1157699 in the calcitonin receptor-like receptor ( CRLR ) gene with primary angle closure ( PAC) in a Han Chinese population.?METHODS: All samples, involved 232 PAC cases and 306 controls, were obtained from an epidemiologic survey conducted in Funing, Jiangsu Province, China. Genotyping were carried out by TaqMan-MGB probe using the real time quantitative polymerase chain reaction system to study the relationship between SNP of rs1157699 in CRLR gene and PAC.?RESULTS: The prevalence of CRLRrs1157699 genotype was 67.4%, 30.0%, 2.6% for CC, CT, TT in cases, and 71.3%, 27.0%, 1.7% in controls respectively.There was no difference between the two groups in the distribution of genotype and allele frequencies of rs1157699 (P>0.05).?CONCLUSION:Our results do not support a significant role for rs1157699 in CRLR with PAC.

Agroinfiltration is a newly developed plant transient gene expression technique, which is simple, rapid and reproducible. It has been widely used in analyses of foreign gene expression, hypersensitive reaction, gene silencing, promoter activity and identification of new disease-resistance genes. In this paper, we describe the principle and the operation procedure of Agroinfiltration and its application in diverse aspects of plant molecular biology research. Our experiences in modification of the Agroinfiltration technique are also provided.
Agrobacterium tumefaciens ; genetics ; Genetic Vectors ; genetics ; Plants ; genetics ; Plants, Genetically Modified ; Research Design

In order to effectively remove the invalid impurities in Tongan injection, optimize the optimal parameters of the impurity removal technology of liquid mixing process, in this paper, taking Tongan injection as the research object, with the contents of celandine alkali, and sinomenine, solids reduction efficiency, and related substances inspection as the evaluation indexes, the removal of impurities and related substances by the combined process of refrigeration, coction and activated carbon adsorption were investigated, the feasibility of the impurity removal method was definited and the process parameters were optimized. The optimized process parameters were as follows: refrigerated for 36 h, boiled for 15 min, activated carbon dosage of 0.3%, temperature 100 degrees C, adsorption time 10 min. It can effectively remove the tannin, and other impurities, thus ensure the quality and safety of products.
Adsorption ; Charcoal ; chemistry ; Drug Compounding ; instrumentation ; methods ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Quality Control

Objective:To observe the effect of oral administration of dietary fiber (DF) on flatus and defecation promotion and abdominal distension reduce,as well as the impact on exclusive breastfeeding and adverse reactions.Method:A total of 80 pregnant women who received repeated cesarean section at obstetrics department in Shanxi Provincial People's Hospital(2016.12 ～ 2017.03) were randomly divided by computer into two groups.DF group included 40 cases,and the patients took DF (1 bag,15g,with 200ml warm boiled water) 6h after cesarean section in 10min,and repeated it 8 ～ 12 h after operation.Patients in control group only received 200ml warm boiled water at the same point-in-time.The two groups shared the same dietary guidance,nursing and medical treatment.Bowel sound,flatus and defecation conditions as well as the incidence rates of abdominal distension and exclusive breastfeeding were compared between two groups.The adverse reactions after administration of DF were collected.Result:There were 38(97.4 ％)cases that had bowel sound in 12h pro-operation in DF group.There were 25(62.5 ％) cases that had bowel sound in 12h pro-operation in control group.There were 30(76.9 ％) and 12(30.0 ％) cases had first flatus during 24h pro-operation respectively in DF group and in control group.There were 22 (56.4 ％) and 7 (17.5 ％)cases had first defecation ＜48h pro-operation respectively in DF group and in control group.The differences were all statistically significant (P ＜ 0.01).In the comparison of gastrointestinal reaction,there were more cases reported abdominal distension(3,7.7％;12,30.0％) and nausea (3,7.7％ ∶10,25.0 ％),with statistical significances (P ＜ 0.05).There was no statistical significance (1,2.6 ％ ∶ 4,1.0 ％,P ＞ 0.05) in vomiting.Breast feeding rate was no statistical significance (56.％ ∶52.5 ％,P ＞ 0.05).One case of DF group discovered adverse reactions,such as nausea and vomiting.Conclusion:Oral administration of soluble DF significantly promoted the flatus and defecation after repeated cesarean section,however,patients with intestinal irritability need to be paid more attention to the adverse reactions,such as nausea and vomiting.

Objective To analyze the related factors with prognosis in patients with serous ovarian adenocarcinoma and to set up a prognostic model of serous ovarian adenocarcinoma.Methods The clinical, pathological and follow-up data of 104 cases with serous ovarian adenocarcinoma were retrospectively analyzed.Kaplan-meier univariate analysis was used to screen the prognostic factors;COX univariate and multivariate analyses were used to determine the risk coefficient of each factors and different layers in each factor.Pearson rank correlation was used to reject the influence of different factors with each other.And the prognostic model of serous ovarian adenocarcinoma was set up based on the result of the above study,which could be used to deduce the survival probability of patients with serous ovarian adenocarcinoma.Results International Federation of Gynecology and Obstetrics(FIGO)stage(P=0.0029),histological grade(P= 0.0054),residual disease(P=0.0000),metastasis of lymph nodes(P=0.0000)and chemotherapy(P= 0.0000)were the related factors of prognosis in patients with serous ovarian adenocareinoma,of which FIGO stage was the most important one,followed sequentially by histological grade,metastasis of lymph node, residual disease and chemotherapy(the independent risk coefficient of each factor was 1.3392,0.9206, 0.7071,0.6004,0.4985 in sequence).We set up a prognosis model according to the prognostic index of each factors.The effect of chemotherapy and residual disease on prognosis could be quantified by this model, and the higher the score,the lower the survival probability of patients.Condusions FIGO stage, histological grade,residual disease,metastasis of lymph nodes and chemotherapy are important prognostic factors of serous ovarian adenoearcinoma.This model can be used to estimate the prognosis of patients with serous ovarian adenoearcinoma,and the effect of both chemotherapy and residual disease on the prognosis could be quantified by the model.

Adult ; Amyloidosis ; diagnosis ; pathology ; Female ; Humans ; Liver Diseases ; diagnosis ; pathology

Objective To investigate the prevalence of uncorrected refractive errors among urban population aged 50 years and above in Ftming county,Jiangsu province.Methods Survey research was conducted and randomly cluster sampling was used to select individuals aged ≥50 years for visual acuity test and eye examination in Funing county,Jiangsu province.The criteria of uncorrected refractive errors in this study was defined as an improvement of at least 0.2 log MAR (equivalent to 2 lines) in the best corrected visual acuity with the base vision ＜ 0.5 log MAR in daily life.The quantitative data were expressed as mean ± standard deviation,and t-test was used for comparison between groups,and while the count data were expressed as rate or composition ratio,and the x2 test was adopted for comparison between the groups.Logistic regression was used to examine the effect of possible factors (i.e.age and gender) on the prevalence of uncorrected refractive errors.Results A total of 6145 persons aged 50 years and above were enumerated and 5947 (96.8％) participants were examined,of whom 2388 had uncorrected refractive errors,with the prevalence of 40.2％.The prevalence of uncorrected refractive errors for myopia only,hyperopia only,astigmatism,and for hyperopia and astigmatism were 84.4％,84.2％,64.1％ and 100％,respectively.Moreover,the higher prevalence of uncorrected refractive errors presented in elderly person (OR =1.07,P ＜ 0.00l) and female (OR =1.38,P ＜ 0.001),and education was a protective factor for junior high school (OR =0.74,P =0.003) and high school (OR =0.55,P ＜ 0.001).Conclusion Uncorrected refractive errors presented high prevalence in rural population aged 50 years and above in Funing county,Jiangsu province,which are the leading cause of visual impairment.

OBJECTIVETo investigate the effects of tanshinone IIA (TA, one of the active components of Salvia miltiorrhiza Bge), on the proliferation of cultured rat vascular smooth muscle cells (VSMC), and to clarity the probable mechanism.

METHODCell culture technique was used in vitro and treated with or without 10% FBS. The inhibitory effect of TA on proliferation of VSMC was observed by cell count, MTU metabolism measuring and BrdU incorporation assay. Flow cytometry was performed to track cell cycle progression. Western bolts were performed to evaluate ERK1/2 activity. The expression of c-fos was examined by RT-PCR.

RESULTThe results of cell number, MTU assay and BrdU incorporation showed that TA cound significantly inhibit 10% FBS induced proliferation in a dose-dependent manner. Flow cytometry (FCM) analysis indicated that the G5/G1 phase fraction ratio of TA group was higher than that of 10% FBS group, while the S-phase fraction ratio was lower than that of 10% FBS group. Western blot's results displayed that TA inhibited the phosphorylation of ERK1/2, and in accordance with this findings. TA could decrease the early elevation of c-fos expression.

CONCLUSIONThese results suggest that TA can inhibit 10% FBS induced the proliferation of VSMC. This effect may be related to its blocking VSMCs cell cycle in G0/G1 phase, inhibiting of the ERK1/2 activity, and decreasing the expression of c-fos.

Animals ; Aorta ; cytology ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Diterpenes, Abietane ; Down-Regulation ; Gene Expression ; drug effects ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Phenanthrenes ; pharmacology ; Proto-Oncogene Proteins c-fos ; genetics ; metabolism ; Rats

The structure analysis of porcine hemoglobin alphabeta dimer and the calculation of solvent accessible surface of the amino acids showed the epsilon-amino groups of the lysine are suitable for modification by polyethylene glycol (PEG). The modification of the lysine residues will not affect the carring oxygen capacity of Hb. Three types of linker have been designed to connect PEG and porcine hemoglobin. The lysines between porcine and bovine hemoglobin (pHb and bHb) are highly conserved, but the solvent accessible surface of conserved lysines are different. These suggested that the properties of homologous proteins are similar in pHb and bHb, but the characteristic derived from the homology analysis will be deviated from the actual status. The results of molecular dynamics simulation suggested that the chemical modified porcine hemoglobin would be no immunogenicity.