ObjectiveTo investigate glutathione S-transferase GSTO 1 Glu155△Glu genetic polymorphism and risks of arsenic poisoning caused by coal-burning in Guizhou.Methods GSTO1 Glu155 △Glu gene polymorphism was analyzed by polymerase chain reaction-with confronting two-pair primers among one hundred and thirty arsenic poisoning patients and one hundred and thirty healthy controls.The results were verified by DNA sequencing.The association between different genotypes and arsenic poisoning was analyzed by unconditional Logistic regression model.ResultsThe results of Glu/Glu and Glu/△Glu genotype detected by this method were consistent with those of DNA sequencing.The frequencies of GSTO1 Glu/Glu genotype and Glu/△Glu genotype were 94.85％(92/97) and 5.15％(5/97) in the patients,99.15％(117/118) and 0.85％(1/118) in the controls,respectively.The difference was statistically significant(x2 =3.896,P ＜ 0.05).△Glu/△Glu genotype was not found in both patients and controls.After age and sex adjusting,GSTO1 Glu155 △Glu polymorphism was found to be a risk factor of arsenic poisoning [odds ratio (OR) =1.85,95％ confidence interval (CI):1.39 - 17.48].ConclusionsThe study finds that GSTO1 Glu 155 △ Glu polymorphism is associated with risk of arsenic poisoning.The relationship between them should be further studied through increasing sample size.

Objective To explore the biological characteristics and karyotype of bone marrow-derived mesenchymal stem cells(MSCs)in patients with systemic lupus erythematosus(SLE)and hematopoietic sup- port of MSCs.Methods MSCs were isolated from bone marrow of 11 SLE patients and 6 healthy controls by density centrifugation and adhesive culture in vitro.The surface markers were detected by flow cytometry (FCM).The morphological changes of MSCs were observed in primary and passage cultures.The growth curves were assayed.The karyotype of MSCs was detected by blocking cellular mitosis with colchicines.The MSCs from SLE patients and healthy controls were infused to ICR mice after high-dose chemotherapy.The changes of peripheral blood counts of the mice were recorded.Results Approximately(6～9)?10~9 MSCs from SLE were obtained after 5 passages and their growth was slower than normal controls(P＜0.01).Both groups were positive for CD29,CD44 and CD105,and negative for CD14,CD34,CD45 and HLA-DR.MSCs from SLE had a normal karyotype.MSCs infusions of the two groups were accompanied by no adverse event and the recovery of white blood cell,hemoglobin and platelet count was quicker when compared with the controls(P＜0.05).Conclusion MSCs from SLE have demonstrated abnormalities in expansion in vitro.MSCs from SLE have a normal karyotype.Ex vivo MSCs infusion from SLE patients can support hematopoiesis as normal MSCs.

Abstract? AIM: Through the establishment of penetrating keratoplasty model of rats, to detect the role and its mechanism of immature dendritic cells with IL-10 gene modified.? METHODS: Allogeneic penetrating corneal transplantation in rat model was performed. SD rats were randomly divided into positive control group, GFP-DC group, 8-DC and IL-10-GFP-DC group.At 3d before keratoplasty, the rats were given tail intravenous injection with the same amount of PBS, bone marrow 8-DC ( DC had cultured for 8d ) from donor Wistar rats, GFP-DC after 48h transfection and IL-10-GFP-DC.Rats were observed under slit-lamp for corneal graft cases every day, and recorded rejection index and corneal graft survival time.At 14d after keratoplasty, pathologic and immunohistochemical examinations were performed.?RESULTS:Compared with GFP-DC group and 8-DC group, corneal graft survival time of IL-10-GFP-DC group was significantly longer ( P<0.01 ); at 14d after keratoplasty, corneal opacity, edema, neovascularization and rejection index of IL-10 -GFP-DC group were significantly lower (P<0.01).Pathological examination showed that in the three experimental groups corneal inflammation was lighter than the positive control group without significant central graft neovascularization. Immunohistochemistry showed: compared to the positive control group, GFP-DC group and 8-DC group, CD4+, CD8+, CD25+, IL-2+, NK+and NF-κB+positive cells in IL-10-GFP-DC group were lower(P<0.01).? CONCLUSION: After donor -derived immature dendritic cells pretreated, corneal graft survival was significantly prolonged, successfully induced corneal transplantation tolerance. CD4+, CD8+, CD25+, IL-2+, NK+and NF-κB+positive cells are involved in corneal allograft rejection regulation, IL-10-GFP-DC may reduce CD4+, CD8+, CD25+, IL-2+, NK+and NF-κB+positive cell infiltration, inhibit corneal transplant rejection.

The aimed of this study was to prepare stabilized thiomers to overcome the poor stability character of traditional thiomers. Poly(acrylic acid)-cysteine (PAA-Cys) was synthesized by conjugating cysteine with poly(acrylic acid) and poly(acrylic acid)-cysteine-6-mercaptonicotinic acid (PAA-Cys-6MNA, stabilized thiomers) was synthesized by grafting a protecting group 6-mercaptonicotinic acid (6MNA) with PAA-Cys. The free thiol of PAA-Cys was determined by Ellmann's reagent method and the ratio of 6MNA coupled was determined by glutathione reduction method. The study of permeation enhancement and stabilized function was conducted by using Franz diffusion cell method, with fluorescein isothiocyanate dextran (FD4) used as model drug. The influence of polymers on tight junctions of Caco-2 cell monolayer was detected with laser scanning confocal fluorescence microscope. The results indicated that both PAA-Cys and PAA-Cys-6MNA could promote the permeation of FD4 across excised rat intestine, and the permeation function of PAA-Cys-6MNA was not influence by the pH of the storage environment and the oxidation of air after the protecting group 6MNA was grafted. The distribution of tight junction protein of Caco-2 cell monolayer F-actin was influenced after incubation with PAA-Cys and PAA-Cys-6MNA. In conclusion, stabilized thiomers (PAA-Cys-6MNA) maintained the permeation function compared with the traditional thiomers (PAA-Cys) and its stability was improved. The mechanism of the permeation enhancement function of the polymers might be related to their influence on tight junction relating proteins of cells.
Acrylic Resins ; chemistry ; Actins ; metabolism ; Animals ; Caco-2 Cells ; Cysteine ; chemistry ; Dextrans ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; Glutathione ; Humans ; Intestinal Absorption ; Intestinal Mucosa ; drug effects ; Nicotinic Acids ; chemistry ; Rats ; Sulfhydryl Compounds ; chemistry

A two-step method for the purification of blocking-type anti-human CD80 monoclonal antibody 4E5 from mouse ascites was developed using anion exchange and gel filtration in combination. The ascites was first purified by anion exchange after centrifugation and filtration. The experimental parameters of sample loading and elution were optimized. The optimized loading condition was pH 8.0,50 mmol/L Tris-HCl and satisfactory results were obtained using a 0~0.5mol/L NaCl step elution. The fraction containing the protein of interest was directly loaded on gel filtration column and eluted using a 20 mmol/L phosphate buffer at pH 7.2. The purity of the obtained monoclonal antibody was up to 95% with a recovery of 61%. The purity of mAb could efficiently inhibit the growth of Daudi cells. The amplification of the method was also studied using a Bio-Scale Q5 column and the result was satisfied.

Objective To investigate histopathological changes and expression of integrin ?1 of sternomastoid muscle,and probe the mechanism and significance during disease process in congenital muscular torticollis(CMT).Methods Histopathological changes of sternomastoid muscle section stained with HE and Gomori silver staining were observed and the expression of integrein ?1 with immunohistochemistry was detected,and the expressive quantity and distribution with image analysis system was quantitive analyzed.Results 1.With light microscopy observation,the results showed that the fibrous degeneration of sternomastoid muscle could be summed up 2 kinds: A category displayed the myocytes atrophyed,and there were lots of connective tissue hyperplasy around myocytes,and the direction of fibrous arrangement was disordered,meanwhile there were lots of vessels and nervers hyperplasy,and eventually the myocytes shrank back and disappeared.B category displayed that the structure of cross striation or sarcomere disappeared or changed,and myocytes could maintained the outline and the sarcolemma were integrated,and then fibrous pathological changes of myocyte took place,and there were lots of fibroblast-like that had much more enations between fiber bundles.With Gomori silver staining,the major changes of fibrotic sternomastoid muscle showed that there were lots of collagenous fibers hyperplasy.The arrangements of collagenous fibers were disordered in A category and were well-arranged in B ca-tegory.2.With immunohistochemistry,the results showed the expression of integrin ?1 was weak positive in normal control group(125.7?5.167).In diseased groups,the results showed 3 different extents:the expression of integrin ?1 displayed stronger positive in A category myocytes(30.15?6.543),and the level of expression was significantly different from normal controls(P0.05).Conclusions The fibrous pathological changes of sternomastoid muscle are a complicated and gradually process,which may has different mode,and ingetrin ?1 may participated the process of pathological changes.

OBJECTIVE To investigate how MLKL functions on the membrane and explore its electrophysiological characters and structure. METHODS The full-length human MLKL were expressed in SF21 cells and purified using glutathione-sepharose affinity chromatography. The currents of purified MLKL proteins were recorded in avoltage-clamp mode using a Warner BC-535 bilayer clamp amplifier. The currents were digitized using pCLAMP 10.2 software. HEK293 cells were cultured and transfected with MLKL plasmid. Cell viability was examined using the CellTiter- Glo Luminescent Cell Viability Assay kit. RESULT MLKL forms cation channels that are permeable preferentially to Mg2+ rather than Ca2+ in the presence of Na+ and K+. Moreover,each MLKL monomer contains five transmembrane helices:H1, H2, H3 , H5 and H6 of the N-terminal domain which is sufficient to form channels. Finally, MLKL-induced membrane depolarization and cell death exhibit a positive correlation to its channel activity.

Adult ; Anti-HIV Agents ; therapeutic use ; Familial Primary Pulmonary Hypertension ; HIV ; pathogenicity ; Hepacivirus ; pathogenicity ; Hepatitis B virus ; pathogenicity ; Humans ; Hypertension, Pulmonary ; drug therapy ; virology ; Male ; Sulfonamides ; therapeutic use

The AtTOM1 gene of Arabidopsis thaliana had been shown to be essential for the efficient multiplication of Tobacco mosaic virus (TMV) in A. thaliana. In this study, we cloned an AtTOM1-like gene from Nicotiana benthamiana named as NbTOM1. Sequence alignment showed that NbTOM1 is closely related to AtTOM1 homologues of N. tabacum and Lycopersicon esculentum with 97.2% and 92.6% nucleotide sequence identities, respectively. Silencing of NbTOM1 by a modified viral satellite DNA-based vector resulted in complete inhibition of the multiplication of TMV in N. benthamiana. The result suggests that inhibition of NbTOM1 via RNA silencing is a potentially useful method for generating TMV-resistant plants.
Arabidopsis Proteins ; genetics ; Cloning, Molecular ; Membrane Proteins ; genetics ; Plant Proteins ; antagonists & inhibitors ; genetics ; RNA Interference ; RNA, Plant ; RNA, Viral ; Sequence Homology ; Tobacco ; metabolism ; virology ; Tobacco Mosaic Virus ; physiology ; Virus Replication

OBJECTIVETo investigate the changing laws of serum high mobility group box 1 protein (HMGB1) in septic rats and intervention effect of Xuebijing on it.

METHODSLipopolysaccharide (LPS) (5 mg/kg BW) was intravenously injected into the tail vein of healthy male Wistar rats to prepare the sepsis rat model. In Experiment 1: 50 Wistar rats were randomly divided into three groups, i.e., the normal group (A, n=10); the LPS model group (B, n=10), the LPS +Xuebijing treatment group (C, n=30). Rats in the C group were further divided into three subgroups, i.e., 2 h before LPS injection (group C1), 2 h after LPS injection (group C2), and 8 h after LPS injection (group C3), 10 in each group. Blood samples were collected from the caudal vein to detect serum HMGB1 levels by Western blot at 4, 12, 24, 48, and 72 h after LPS injection. Experiment 2: 30 Wistar rats were equally divided into the LPS model group (D) and the LPS + Xuebijing treatment group (E), 15 in each group. They were treated as rats in the B group and the C1 group respectively. Five rats were sacrificed at 12, 24, and 48 h after LPS injection in the two groups. Blood as well as the tissue samples were harvested to measure such indices as ALT, AST, Cr, and BUN, as well as pathological changes of liver, lung, and kidney.

RESULTS(1) Compared with the A group, serum HMGB1 levels were higher at various time points in the B group (P < 0.05). Compared with the B group, serum HMGB1 levels at 12,24,48, and 72 h decreased in the C1, C2, and C3 groups. Besides, the decrease was more obvious at 24 h and 48 h.The decrement in the C3 group was less than that in the C1 and C2 groups (P < 0.05). (2) In the D group, ALT, AST, Cr, and BUN were significantly higher than those in the A group and reached the peak at 24 h (P < 0.05). Compared with the E group, AST, Cr, and BUN at 24 and 48 h, and ALT at each time point decreased significantly in the E group (P < 0.05). (3)The results of pathological section of liver, lung, and kidney showed local congestion and hemorrhage, cell edema/necrosis/degeneration, infiltration of inflammatory cells, damage of characteristic structures and so on; particularly serious lesion occurred at 24 and 48 h in the D group. The microscopic lesion was obviously alleviated in the E group than in the D group at corresponding time points.

CONCLUSIONSThe serum HMGB1 levels increased in septic rats, with late occurrence of peak value and longer duration of the high value. HMGB1 played an important role in excessive inflammatory response and multiple organ dysfunction. Xuebijing could reduce the serum levels of HMGB1, improve biochemical parameters, and attenuate severe inflammatory response of liver, lung, and kidney tissues in septic rats. Besides, the earlier use, the better effect obtained.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; HMGB1 Protein ; blood ; Male ; Rats ; Rats, Wistar ; Sepsis ; blood ; drug therapy