Objective To investigate the relationship between the microsatellite polymorphism of cytotoxic T lymphocyte associated antigen-4 (CTLA-4) gene and inflammatory bowel disease(IBD) patients in Zhejiang province. Methods One hundred and eighteen IBD patients without consanguinity (99 patients with ulcerative colitis, 19 patients with Crohn's disease) and 140 healthy controls were studied. The (AT) repeats in the 3' untranslated region of exon 4 of CTLA-4 gene were amplified by allele-specific PCR. The amplified products were electrophoresed with 12% polyacrylamide gel, followed by silver staining. Results Twenty alleles were found in IBD patients and healthy controls. The frequency of 122 bp allele was higher in patients with ulcerative colitis (P=0.0001/Pc=0.0025, OR= 11.393, 95% CI:2.574-50.429) and Crohn's disease (P=0.0003/Pc=0.0050, OR=21.061, 95% CI:3.927-112.94) than that in healthy controls. Conclusion CTLA-4 gene microsatellite polymorphism was obviously associated with IBD in Zhejiang province.

Objective To explore the effect of different cold ischemia (CI)times on the patterns of intra-graft T cell activation gene-3 (TCA-3) expression early after isogeneic half liver transplantation (PLT)in mice. Methods The models of C57BL/6 full-size(FS) and PLT were established.The CI time was 1,4 and 8 h.Specimen were collected at 4 and 24 h post-reperfusion.Ribonuclease protection assays (RPA)was used to evaluate serial mRNA expression of the TCA-3 chemokine in all mice.Histopathology was used to examine cold ischemia injury in the liver grafts. Results A total of 45 control and 30 PLTs were performed.The survival rate at 7 day after control and PLT was 100％.Quantitative analysis demonstrated the levels of TCA-3 mRNA expression were low at 4 h after reperfusion in control group with 1,4,8 h CI.The levels of TCA-3 significandy increased at 4 h after reperfusion in PLT group with CI of 1,4,8 h and the difference between the two groups was statistically significant ( F =7.41,P ＜ 0.05 ). TCA-3 mRNA expression significantly decreased at 24 h after reperfusion in two groups. But the difference was not statistically significant ( P ＞ 0.05 ). Histopathology showed severer cold ischemia injury in PLT grafts compared with control grafts. Conclusions The expression of TCA-3 significandy increased early after PLT and played an important role in cold ischemia injury.TCA-3 could be used as the early therapeutic target for reducing ischemia injury in PLT grafts.

Calcaneal fracture is a kind of common injury of foot. Calcaneal fracture could be treated with conservative therapy, open reduction and internal fixation (ORIF), minimal invasive poking reduction and Kirschner wire fixation, arthrodesis or external fixation and other treatment methods. Open reduction and internal fixation as the main treatment method of calcaneus fracture, especially for intra-articular calcaneal fractures, as Sanders type II and III fractures, could gain satisfactory effect. But whether bone grafting should be taken is controversial in Sanders type IV fractures, because soft tissue complications are disadvantages of ORIF. The treatment of minimally invasive percutaneous poking reduction and internal fixation for calcaneal fractures has been paid more and more attention. Because of the differ of the classification of calcaneal fractures, indications of poking reduction and fixation methods, efficacy evaluation standard of choose, the clinical efficacy is different. Because of the complex treatment process, various treatment methods have their own advantages and disadvantages. Through consulting relevant literatures of domestic and abroad in recent years, comprehensive analysis is made and new progress of the calcaneal fractures treatment is summarized.
Calcaneus ; injuries ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Minimally Invasive Surgical Procedures

Cell Membrane ; metabolism ; Gastrointestinal Stromal Tumors ; diagnosis ; metabolism ; Humans ; Immunohistochemistry ; Proto-Oncogene Proteins c-kit ; analysis

Objective To assess the application of three-dimensional digital subtraction angiography (3D-DSA) in evaluation of ruptured intracranial aneurysm after clipping and to discuss the different variable use of vol-ume rendering(VR), gradient rendering (GR) and maximum intensity projection (MIP). Methods From January 2011 to December 2012 , 88 patients with 92 ruptured intracranial aneurysms were treated with clipping using titani-um clips in our hospital and followed up by both 2D-DSA and 3D-DSA. Residual aneurysms , Clips place, clips and parent arteries and stenosis of parent arteries were evaluated by volume rendering (VR), gradient rendering (GR) and maximum intensity projection (MIP). Results Among 92 clipped aneurysms, 23 residual aneurysms were found by 3D-DSA. Residual aneurysms were recorded according to the Sindou grade: 15 of gradeⅠ, 3 of gradeⅡ, 4 of grade Ⅲand 1 of grade Ⅳ. Three patients of grade Ⅲand 1 of grade Ⅳwith residual aneurysms were retreated by clipping or coiling, and 1 patient of grade Ⅲ was dead with rupture of residual aneurysm. The clips and number of clips were clearly visualized , and relationship between the clips and the aneurysms was well demonstrated by VR, GR and MIP images. VR, GR images showed the remnants clearly. Three-dimensional digital subtraction angiography did not showed accurate details of the stenosis of parent arties which required an analysis of 2D-DSA. Conclusion Three-dimensional digital subtraction angiography can be used for definite evaluation of resid-ual aneurysms after clipping, especially by VR, GR images. It is helpful to manage the residual ruptured aneurysms.

AIM:To evaluate the application and effect of bandage contact lens in pterygium excision combined with conjunctival transplantation (CAT).METHODS:In a prospective, randomized, controlled clinical study, 110 patients (110 eyes) diagnosed with primary pterygium were collected in PLA NO.474 Hospital from January 2015 to January 2016.The 110 patients enrolled in the study in turn, and divided into two groups by the odd and even number.The odd number divided into bandage contact lens group (CAT + bandage contact lens, n=55), while the even number divided into control group (CAT, n=55).Visual analog scale (VAS) and corneal irritation were evaluated on 1, 3 and 7d after operation.Cornea fluorescent staining testing was carried out on 3d after operation.Following-up all the patients with 1a at least observed the recurrence of pterygium.RESULTS:The score of VAS in bandage contact lens group less than that in control group on 1d (4.13±2.06 vs 5.80±1.93, t=4.391, P<0.001) and 3d (2.09±1.36 vs 3.65±1.65, t=5.422, P<0.001) after operation, while there was no significant difference between two groups on 7d (t=1.295, P=0.198) after operation.The corneal irritation in bandage contact lens group less than that in control group on 1d and 3d after operation (P<0.05), while there was no significant difference between two groups on 7d after operation (P=0.052).Cornea fluorescent staining testing area in bandage contact lens group was less than that in control group on 7d after operation (0.33±0.37mm2 vs 2.73±2.21mm2, t=7.921, P<0.01).There was no significant difference in recurrence rate between two groups after 1a operation (P=1.000).CONCLUSION:Bandage contact lens could significantly release pain and corneal irritation, promote the healing of the corneal epitheliums in the defected area, and increase the postoperative comfort level in patients after operation.

Porcine reproductive and respiratory syndrome virus (PRRSV) has been recognized as one of the most important pathogens of pigs throughout the world. In 2006, more than 10 provinces of China have experienced an epizootic outbreak of pig diseases characterized by high fever, reddened skin and high morbidity and mortality. From June 2006 to April 2007, we have investigated some clinical samples in Hubei province by RT-PCR and cloned several major genes, N, GP5 and NSP2 gene, shown in this study. Phylogenetic analysis of these genes revealed that the highly pathogenic PRRSV variant, ZB, was responsible for 2006 emergent outbreak of pig disease in Hubei province similar with those variants isolated from other provinces in China in 2006, and belongs to the NA-type PRRSV. In the PRRSV variants, the N and GP5 shear about 90% identity with prototypic ATCC VR-2332 and some typical NA-type Chinese isolates, except the 2850bp NSP2 gene (only shares 65% identity with ATCC VR-2332). But they all shear more than and 97% identity with other highly pathogenetic Chinese PRRSV strains. Additionally, there are extensive amino acid (aa) mutations in the GP5 protein and 2 deletions in the Nsp2 protein when compared with the previous isolates. Most of the variants found in 2006 epizootic outbreak of pig diseases in China were the farthest variants from the typical NA-type PRRSV in phylogenetic distance, and these diversities may be responsible for the differences in the pathogenicity observed between these variants and original Chinese PRRSV strains.

OBJECTIVETo study the effect of antizyme 1 (ZA1) gene transfection on the cell proliferation, cell cycle and apoptosis of human neuroblastoma SH-SY5Y cells in vitro.

METHODSThe recombinant eukaryotic expression vector pAZ1m was constructed by cloning mutant AZ1 gene into the vector pEGFP-N1, and subsequently transfected in SH-SY5Y cells. The transfected cell proliferation was examined using MTT assay, and the changes in cell cycle and apoptosis were assayed using flow cytometry analysis. RT-PCR was performed to measure cyclin D1 and caspase-3 mRNA expressions, Western blotting carried out to examine cyclin D1 protein expression, and the changes in caspase-3 activity were detected using a caspase-3 detection kit.

RESULTSAZ1 gene transfection significantly inhibited the proliferation of SH-SY5Y cells, causing cell cycle arrest at G0/G1 stage and down-regulated cyclin D1 and up-regulated caspase-3 expressions. Obviously increased caspase-3 activity was also observed in the transfected cells.

CONCLUSIONExogenous AZ1 gene transfection can inhibit the proliferation and cause cell cycle arrest of SH-SY5Y cells by down-regulating cyclin D1 expression. Up-regulated caspase-3 expression resulting from AZ1 gene transfection may induce apoptosis of the neuroblastoma cells.

Apoptosis ; Caspase 3 ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Down-Regulation ; Genetic Vectors ; Humans ; Neuroblastoma ; metabolism ; Proteins ; genetics ; Transfection ; Up-Regulation

OBJECTIVETo provide the basis for clinical acquired immunodeficiency syndrome (AIDS) surveillance and to avoid cross infection in hospital, we study the infection status of AIDS in Shandong province.

METHODSThe fourth-generated Akzo's ELISA kit and the fourth generated Immunoluminometric detection reagent were used for HIV antibody screening for 399 303 cases of both inpatients and outpatients from Jan. 2003 to Dec. 2011. Beijing WanTai ELISA kit and Se-marked rapid detection reagent were used for re-detection, and the positive samples were sent to the local CDC for confirming test by Western Blot.

RESULTSThe HIV-1 antibody detection results of 129 (0. 3230 per thousand) patients were confirmed to be positive, including 54 (0. 1352 per thousand) cases of outpatients and 75 (0. 1878 per thousand) cases of inpatients. HIV infection rates in outpatients from 2003 to 2011 were 0.050 per thousand, 0.030 per thousand, 0.111 per thousand, 0.120 per thousand, 0.124 per thousand, 0.113 per thousand, 0.148 per thousand, 0.201 per thousand, 0.2152 per thousand; and that in inpatients were 0. 150 per thousand, 0.089 per thousand, 0.138 per thousand, 0. 144 per thousand, 0. 104 per thousand, 0. 132 per thousand, 0. 197 per thousand, 0. 329 per thousand, 0. 313 per thousand respectively. Among these inpatients, there were 61 cases of medical patients and 14 cases of surgical patients, and most were youths and farmers.

CONCLUSIONSHIV infection rate was increasing year by year. Most inpatients whose HIV-1 antibody was positive were in the phase of AIDS. Therefore, it's very necessary to execute routine testing for inpatients and outpatients who need special examination for early diagnosis of HIV infection

Acquired Immunodeficiency Syndrome ; epidemiology ; immunology ; virology ; Adolescent ; Adult ; Aged ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Female ; HIV Antibodies ; analysis ; HIV Infections ; epidemiology ; immunology ; virology ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the effect of Astragalus Injection solution on rat hepatic stellate cells (HSC) and hepatic fibrosis.

METHODSHSCs of rats were incubated with various concentrations of Astragalus Injection solution (0 mg/ml, 25 mg/ml, 50 mg/ml, 100 mg/ml, 200 mg/ml, 400 mg/ml) for 24, 48 and 72 hours. Cell proliferation was detected with 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphennyltetrazolium bromide (MTT) colorimetric assay. Cell cycle was detected with flow cytometry. Cell apoptosis was detected with acridine orange/ethidium bromide (AO/EB) fluorescent staining and flow cytometry. In vivo, rats were randomly allocated into a normal control group, a model control group and an Astragalus Injection group. Astragalus Injection (800 mg.kg-1.d-1) was administered to rats of the Astragalus Injection group. Rats of the model control group received saline. Serum concentrations of hyaluronic acid (HA) and laminin (LN), hepatic tissue activity of superoxide dismutase (SOD), and hepatic tissue contents of malondialdehyde (MDA) were measured in these groups at 8 weeks. Hepatic tissue expression of LN was assessed by using immunohistochemistry. The pathological changes of hepatic tissues were examined by hematoxylin-eosin (HE) and van Gieson (VG) staining of their histological slides.

RESULTSIn vitro, compared with the 0 mg/ml group, the proliferation of HSCs in other concentration groups was significantly inhibited by Astragalus Injection solution in a dose and time dependent manner, the cell proliferation cycle of HSCs was blocked in the G2-M phase, there was no apoptosis of HSCs in AO/EB fluorescent staining and flow cytometry. In vivo, compared with rats of the model control group, the rats of the Astragalus Injection solution treated group had remarkably decreased serum HA and LN levels (114.3+/-25.6) microg/L vs (85.6+/-37.3) microg/L and (78.8+/-11.7) microg/L vs (66.8+/-17.6) microg/L, P < 0.05, and liver MDA level (3.7+/-0.4) micromol/g protein vs (2.4+/-0.2) micromol/g protein, P < 0.01, but had increased activity of liver SOD (49.6+/-5.7) NU/mg protein vs (75.9+/-5.9) NU/mg protein, P < 0.01. Microscopic studies revealed that the livers of rats receiving Astragalus Injection solution showed decreases in fibrosis and in expression of LN.

CONCLUSIONSAstragalus Injection solution has an inhibitive effect on experimental hepatic fibrogenesis. The mechanisms of its effects might possibly be associated with its antioxidant activity, expression of decreasing LN and its inhibition of HSCs proliferation.

Animals ; Astragalus membranaceus ; Cells, Cultured ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hepatocytes ; pathology ; Injections ; Liver Cirrhosis, Experimental ; drug therapy ; pathology ; Male ; Phytotherapy ; Random Allocation ; Rats ; Rats, Wistar