Adult ; Blast Injuries ; therapy ; Explosions ; Humans ; Male ; Multiple Trauma ; therapy

Objective　To observe some biological features of bone marrow mesenchymal stem cells and explore the best conditions for isolatin g and culturing in vitro. Methods　Common cell culture techn ique, light and electron microscopy were used to study the effects of the growth , proliferation, morphology of the bone marrow mesenchymal stem cells in differe nt adherent time, concentration of serum and cell density. Results　The best culture condition in vitro for growth was 4-24 hours adherent time, 5%-10% fetal bovine serum, (4-8)×104/ml cell density. The cells were sp indle in shape and had a strong ability of proliferation. The time for cell duplication was 3 to 4 days. The cells showed the characteristics of stem cell s in electron microscope. Conclusion　The best condition for iso lation and culture of bone marrow mesemchymal stem cells was successfully establ ished and some biological features were obserred. It found a base for further in vestigation and using of mesenchymal stem cells.

Heart Ventricles ; physiopathology ; Humans ; Hypertension, Pulmonary ; etiology ; Male ; Middle Aged ; Myocardium ; pathology

OBJECTIVETo explore the effects of Huoxiang Zhengqi liquid (HXZQ) on enteric mucosal immune responses in mice with Bacillus dysenteriae and Salmonella typhimurium induced diarrhea (BSD).

METHODMice were randomly divided into four groups with 10 mice in each group: control group (control), BSD group, Huoxiang Zhengqi liquid treated BSD groups at high dosage and low dosage (HXZQ high, HXZQ low). HXZQ was administrated from the day of diarrhea induction at dosage of 5.21 g kg(-1) and 0.52 g kg (-1) respectively. Peyer's patch and periphery lymphocytes were prepared for flow cytometry, and level of TNF-alpha in periphery and enteric tissue homogenate were determined with ELISA. Student's t-test was used for statistics.

RESULTMice in BSD group started showing continuous diarrhea at the day of induction till the fourth day when the mice were sacrificed. Diarrhea in the mice of HXZQ high and low groups lasted for 36 and 54 h respectively. There were more CD4+ and CD8+ cells in periphery, less CD4+ cells in peyer's patch in BSD mice comparing to normal mice. In peyer's patch, there were more CD8+ cells in mice in HXZQ high and low groups and more CD4+ in mice in HXZQ high group. Higher level TNF-alpha in periphery and intestinal tissue homogenate in BSD group were observed. Mice in HXZQ high group showed the decreased level TNF-alpha in periphery and enteric tissue homogenate.

CONCLUSIONThe immune regulation on peyer's patch CD4+ and CD8+ cells and suppression on TNF-alpha level in enteric homogenate might partially explain the effect of HXZQ on improvement of BSD.

Animals ; CD4-CD8 Ratio ; Colon ; immunology ; metabolism ; pathology ; Diarrhea ; immunology ; metabolism ; microbiology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Dysentery, Bacillary ; immunology ; metabolism ; microbiology ; Immunity, Mucosal ; drug effects ; Intestinal Mucosa ; immunology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Peyer's Patches ; drug effects ; immunology ; pathology ; Plants, Medicinal ; chemistry ; Random Allocation ; Salmonella Infections ; immunology ; metabolism ; microbiology ; Salmonella typhimurium ; immunology ; Shigella dysenteriae ; immunology ; T-Lymphocyte Subsets ; drug effects ; immunology ; pathology ; Tumor Necrosis Factor-alpha ; blood ; metabolism

AIMTo study the chemical constituents of Sargentodoxa cuneata.

METHODSTo isolate compounds with chromatography technology and to elucidate their structures by spectral analysis.

RESULTSTen phenolics were isolated from Sargentodxa cuneata and their structures were determined as 1-O-(vanillic acid )-6-O-( 3", 5"-dimethoxy-galloyl)-beta-D-glycoside (I), (-)-epicatechin (II), phydroxyphenylethanol ferulate (III), chlorogenic acid (IV), methyl chlorogenate (V), apocynin (VI), vanillic acid (VII), protocatechuic acid (VIII), 3,4-dihydroxy-phenylethanol (IX), tyrosol (X).

CONCLUSIONCompound I is new compound, compound III-VI and VIII-X were isolated from Sargentodoxa cuneata for the first time.

Acetophenones ; chemistry ; isolation & purification ; Chlorogenic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Hydroxybenzoates ; chemistry ; isolation & purification ; Magnoliopsida ; chemistry ; Molecular Conformation ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Vanillic Acid ; analogs & derivatives ; chemistry ; isolation & purification

To combine the fibrinolytic with anticoagulant activities for therapy of thrombotic deseases, a fusion protein made of tissue-type plasminogen activator (t-PA) and hirudin was constructed and expressed in chia pastoris. To improve thrombolytic properties of t-PA and reduce bleeding side effect of hirudin, FXa-recognition sequence was introduced between t-PA and hirudin molecules.The anticoagulant activity of hirudin can be target-released through cleavage of FXa at thrombus site. t-PA gene and hirudin gene with FXa-recognition sequence at its 5'-terminal were obtained by RT-PCR and PCR respectively. The fusion protein gene was cloned into plasmid pIC9K and electroporated into the genome of Pichia pastoris GS115. The expression of fusion protein was induced by methanol in shaking flask and secreted into the culture medium. Two forms of the fusion protein, single-chain and double-chain linked by a disulfide bond (due to the cleveage of t-PA at Arg275-Ile276), were obtained. The intact fusion protein retained the fibrinolytic activity but lacked any anticoagulant activity. After cleavage by FXa, the fusion protein liberated intact free hirudin to exert its anticoagulant activity. So, the fusion protein is a bifunctional molecule having good prospect to develop into a new targeted therapeutic agent with reduced bleeding side effect for thrombotic diseases.
Animals ; Cloning, Molecular ; Electroporation ; Hirudins ; biosynthesis ; genetics ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Plasminogen Activator ; biosynthesis ; genetics

Objective To investigate the anti-fibrosis action of the therapy of tonifying kidney and removing stasis(shortened to Bushen Quyu method)on moderate-to-severe intrauterine adhesion and its influence on extracellular matrix degradation related factors in menstrual blood.Methods A total of 124 patients with moderate-to-severe intrauterine adhesions of kidney deficiency and blood stasis syndrome who were to undergo transcervical resection of adhesion(TCRA)were randomly divided into a study group and a control group,with 62 patients in each group.Both groups were given TCRA treatment.After the operation,the control group was treated with Progesterone + Estradiol Valerate Tablets,and the study group was treated with the combination of the Bushen Quyu method by using modified Guishen Decoction based on the treatment of the control group.The treatment covered 3 menstrual cycles.The changes in menstrual status,transvaginal color Doppler sonography parameters,scores of kidney deficiency and blood stasis syndrome,and scores of uterine adhesion in the two groups were observed before and after the treatment.Moreover,the two groups were observed before and after treatment in the changes of vascular endothelial growth factor(VEGF),transforming growth factor β1(TGF-β1),platelet-derived growth factor(PDGF),and connective tissue growth factor(CTGF)as well as the mRNA expression levels of matrix metalloproteinase 9(MMP-9),matrix metalloproteinase inhibitory factor 1(TIMP-1),phosphatidylinositol-3-hydroxykinase(PI3K)and collagen type I A1 protein(COL1A1).After treatment,the clinical efficacy and safety of the two groups were assessed.Results(1)After 3 menstrual cycles of treatment,the total effective rate in the study group was 93.55%(58/62),and that in the control group was 80.65%(50/62).The intergroup comparison showed that the therapeutic effect of the study group was significantly superior to that of the control group(P＜0.05).(2)After treatment,the menstrual flow,menstrual cycle and menstrual period of the two groups were significantly improved compared with those before treatment(P＜0.05),and the recovery of menstrual flow,menstrual cycle and menstrual period in the study group was significantly superior to that in the control group after treatment(P＜0.05).(3)After treatment,the transvaginal color Doppler sonography parameters of uterine blood flow index(FI),pulsatility index(PI),resistance index(RI),uterine cavity volume,and endometrial thickness in the two groups were significantly improved compared with those before treatment(P＜0.05).And the study group had significantly higher FI,uterine cavity volume,and endometrial thickness and had lower PI and RI than the control group(P＜0.05 or P＜0.01).(4)After treatment,the scores of uterine adhesion and the scores of kidney deficiency and blood stasis syndrome in the two groups were significantly decreased compared with those before treatment(P＜0.05),and the above scores in the study group were significantly lower than those of the control group after treatment(P＜0.01).(5)After treatment,the levels of VEGF,TGF-β1,PDGF,and CTGF in the two groups were significantly decreased compared with those before treatment(P＜0.05),and the levels of the above cytokines in the study group were significantly lower than those in the control group after treatment(P＜0.01).(6)After treatment,the relative mRNA expression levels of MMP-9 and PI3K in the menstrual blood of the two groups were significantly higher and the mRNA expression level of COL1A1 was significantly lower than that before treatment(P＜0.05).And the study group had higher mRNA expression levels of MMP-9 and PI3K in the menstrual blood than the control group after treatment(P＜0.05 or P＜0.01).(7)The total incidence of adverse reactions in the control group was 25.81%(16/62)and that in the study group was 16.13%(10/62).The intergroup comparison showed that the difference between the two groups was not statistically significant(P＞0.05).The symptoms of adverse reactions in the two groups disappeared after symptomatic treatment,and there were no adverse reactions related to the medication of Chinese medicine during the study.Conclusion The combination of Bushen Quyu formula combined with western medicine is more effective than western medicine alone for the treatment of patients with uterine adhesions after TCRA.The application of Bushen Quyu formula can further promote the recovery of menstruation in the patients,improve the circulation of blood flow in the endometrium,increase the endometrial thickness,regulate the expression of fibrosis factors,up-regulate the mRNA expressions of PI3K and MMP-9,restore the balance of the extracellular matrix(ECM),and prevent the reoccurrence of intrauterine adhesions.And its clinical medication is safe.

Adult ; China ; epidemiology ; Circoviridae Infections ; epidemiology ; virology ; DNA, Viral ; analysis ; Female ; Genotype ; Hepatitis, Viral, Human ; virology ; Humans ; Male ; Substance Abuse, Intravenous ; virology ; Torque teno virus ; genetics ; isolation & purification

The 109 whole blood samples were collected from HIV-1 infected former blood donors in Henan and Shanxi. The RNA templates were extracted from plasma and used for the full gag gene amplification and sequencing. The sequences were divided into 3 groups according to sampling year. The Entropy software was used to identify the amino acids with composition difference among different groups of amino acid sequences. The results showed that there existed 8 and 13 amino acid sites with the statistical significance difference, respectively, in sequences in year 2004 and 2005, compared to those in 2002. Among them, there existed 5 amino acid sites in two groups. Of 16 amino acid sites, the increasing polymorphism and the decreasing polymorphism along the sampling year were observed in 10 and 6 amino acid sites respectively. Of 10 sites with increased polymorphism, 8 sites were located in the CTL epitopes recognized and presented by the main HLA alleles existed in Chinese population. The 6 sites with decreasing polymorphism all existed in main domains of Gag proteins.
Blood Donors ; China ; epidemiology ; Genetic Variation ; HIV-1 ; genetics ; Humans ; Polymorphism, Genetic ; gag Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVETo describe the incidence trends and pathological characteristics of lung cancer in urban Beijing, China.

METHODSA total of 32 845 medical records of the residents diagnosed as lung cancer in urban Beijing from 1998 to 2007 were retrieved through the cancer registry system of Beijing Cancer Registry. Crude incidence rate, age-specific incidence rate, adjusted incidence rate by world standardized population, annual percentage change (APC) and histological categorized incidence rate by world standardized population were calculated in order to compare the differences of the incidence trends in different time periods, or among different gender and age groups.

RESULTSA total of 32 845 newly diagnosed lung cancer patients between 1998 and 2007 were included in our study. The crude incidence rate was 47.81/100 000 (32 845/68 704 429), increasing by 38.80% from 39.30/100 000 in 1998 to 54.55/100 000 in 2007 with APC at 3.35% in urban Beijing (Z = 9.984, P < 0.001). While it changed to 28.95/100 000 with an APC at 0.27% (Z = 0.846, P = 0.422) when adjusted by world standardized population. For male, the crude incidence rate was 58.28/100 000 (20 342/34 906 580, adjusted rate at 37.03/100 000, APC at 0.38%, Z = 1.008, P = 0.343); while for female, the crude incidence rate was 36.99/100 000 (12 503/33 797 849, adjusted rate at 21.48/100 000, APC at 0.14%, Z = 0.431, P = 0.678). 17 920 lung cancer patients being diagnosed according to histological evidence, accounted for 54.56%. The respective proportion of the patients with histological diagnosis was 43.14% (1095/2538) in 1998 and 65.55% (2641/4029) in 2007, with a 51.95% increase (χ(2) = 859.152, P < 0.001) in decade. In terms of subtypes of lung cancer, the proportion of squamous cell carcinoma decreased annually, from 30.41% (333/1095) in 1998 to 24.16% (638/2641) in 2007; while the proportion of adenocarcinoma increased from 42.83% (469/1095) to 46.80% (1236/2641). As a result, the squamous cell carcinoma to adenocarcinoma ratio declined from 0.71 (333/469) to 0.52 (638/1236) (χ(2) = 50.214, P < 0.001). For women, the ratio declined more significantly and the proportion of the squamous cell carcinoma and adenocarcinoma were 14.77% (925/6262) and 60.83% (3809/6262), respectively in the period between 1998 and 2007.

CONCLUSIONNo significant change was found in the incidence trend of lung cancer after the incidence rate adjusted by world standard population, but the proportion of the subtypes of lung cancer categorized by histological evaluation changed apparently.

Adenocarcinoma ; epidemiology ; pathology ; Adult ; Aged ; China ; epidemiology ; Female ; Humans ; Incidence ; Lung Neoplasms ; epidemiology ; pathology ; Male ; Middle Aged