Objective To discuss the effects of capsular tension ring after phacoemulsification combined with IOL implantation on tilt and decentration of IOL in high myopia patients with cataract by ultrasonic biomicroscope (UBM).Methods A total of 36 cases (40 eyes) with high myopia and cataract underwent phacoemulsification combined with IOL implantation were chosen.The average axial length was 26.88 mm.The patients were divided into implant group (19 eyes,the capsular tension ring was implanted) and control group (21 eyes,the routine surgery was performed).The patients were examined by conventional slit lamp,and the best corrected visual acuity (BCVA)was measured at pre-operation and postoperative 6 months.Tilt and decentration were measured horizontally and vertically,and total tilt and decentration were calculated by geometry method.Results The postoperative BCVA in the two groups were all better than the pre-operation,there was no statistical difference in the preoperative and postoperative BCVA between two groups (all P ＞ 0.05).The horizontal,vertical and total decentration the implant group were (0.15 ± 0.07) mm,(0.30 ± 0.40) mm,(0.11 ±0.02)mm,respectively,which in the control group were (0.26 ± 0.19)mm,(0.32 ±0.60) mm,(0.24 ± 0.97) mm,respectively.The horizontal,vertical and total tilt in the implant group were 0.02° ±0.11°,0.70° ±0.25°,0.21° ±0.74°,respectively,which in the control group were 0.11 ° ± 0.31 °,1.09° ± 0.20°,1.24° ± 0.97°,respectively.There were statistical differences in the horizontal,total tilt and decentration between two groups (all P ＜ O.05),but no statistical difference in the vertical tilt and decentration (P ＞ 0.05).Conclusion The capsular tension ring can stable the IOL position after surgery in high myopia and cataract patients.

Objective To compare clinical efficacy of inverse less invasive stabilization system (LISS) and Gamma nail in treating unstable femoral intertrochanteric fractures combined with lateral wall fractures.Methods Fifty-two patients with intertrochanteric fractures associated with lateral wall fractures followed up for at least 12 months after surgical treatment between June 2010 and June 2013 were included in this retrospective cohort study.According to the different internal fixation,the patients were assigned to inverse LISS group [16 males and 8 females;(62.5 ± 12.4)years] and Gamma nail group [17 males and 11 females;(60.4 ± 18.6)years].According to the AO classification,there were 6 patients with A2.2 type,5 A2.3 type,5 A3.1 type,6 A3.2 type and 2 A3.3 type in inverse LISS group,and there were 4 patients with A2.2 type,7 A2.3 type,9 A3.1 type,5 A3.2 type and 3 A3.3 type in Gamma nail group.Opcration time,total blood loss (intraoperaive plus occult blood loss),hospital length of stay,bone healing,time of commencing full weight-beating and complication incidence were compared between the two groups.Harris hip score was recorded postoperatively.Results All patients were followed up for 24-36 months (mean,30.2 months).Operation time in Gamma nail group was shorter than that in inverse LISS group (P ＜ 0.05),while relatively more blood loss was found in Gamma nail group(P ＜0.05).There were no significant differences in hospital length of stay and bone healing timebetween the two groups (P ＞ 0.05).Patients in Gamma nail groups started earlier full weight-bearing than inverse LISS group (P ＜0.05).Harris score was (86.1-± 12.4)points in inverse LISS group one year after operation,not significantly different from (83.3 ± 11.2) points in Gamma nail group (P ＞ 0.05).Complication of one patient with coxa vara and one bone nonunion in inverse LISS,showing no significant from one patient with screw breakage and one femoral head osteonecrosis in Gamma nail group (P ＞ 0.05).Conclusions Both techniques are effective in treatment of intertrochanteric fractures combined with lateral wall fractures and achieve similar results in function recovery.But inverse LISS is eccentric fixation,so too early weight-bearing should not be over-emphasized.

Objective To construct prokaryotic expression plasmid of human recombinant soluble TNF-related apoptosis inducing ligand(rsTRAIL),then to investigate the effects of rsTRAIL on apoptosis in non-small cell lung carcinoma(NSCLC).Methods The encoding sequence for rsTRAIL was amplified with RT-PCR and cloned into PQE30 vector to establish the prokaryotic expression system.The competent cells of host strain of M15 were transformed by the recombinant plasmid.The expression of the target protein was induced with IPTG and purified by Ni2+-NTA agarose column.rsTRAIL was added in the media of A549 and H460wt cells,then the viability was examined by MTT assay.Apoptosis of H460wt cells was observed under fluorescence microscopy.The apoptotic rate of tumor cells was examined by FACS.Results The cloned fragment of rsTRAIL was 100% consistent with that reported in GenBank.The expressed protein with molecular weight of 21 000 in SDS-PAGE as expected was obtained and recognized by a commercial McAb.The apoptotic rate of H460wt cells after treated with rsTRAIL for 24 h was 43.2%.Cislatin enhanced the effect of rsTRAIL on A549 cells.Conclusion The rsTRAIL is obtained after Ni affinity chromatograph.rsTRAIL has a strong cytotoxic activity against NSCLC and cisplatin may enhance the antitumor effect of rsTRAIL.

Objective:To observe the change of CD4+CD25+regulatory T cells and Th17 cells in mice with experimental anti-phospholipid antibody syndrome ( EAPS ) .Methods: EAPS model was established by immunizing BALB/c mice with recombinant humanβ2 glycoprotein 1 (rhβ2GP1).The levels of serum anti-β2 glycoprotein 1 (anti-β2GP1),anti-cardiolipin antibody (aCA),IL-17,IL-2,IL-6 and TGF-βwere tested by ELISA.The rate of abortion,activated partial thromboplastin time (APTT) and platelet count were also detected.Flow cytometry was applied to detect the percentages of the CD4+CD25+regulatory T cells and Th17 cells in peripheral blood mononuclear cells.Results:Compared with the control group,the levels of anti-β2 GP1,aCA,IL-17,IL-2 and IL-6 were significantly increased,the rate of abortion was increased,APTT time was prolonged and the levels of TGF-βand platelet count were de-creased in model mice (P<0.05).No significant difference was detected of percentage of Treg cells in PBMC at the eighth weeks in model group (P>0.05),but percentage of Treg cells was lower than that in control group after 12 weeks (P<0.05);the percentage of Th17 cells in model group was higher than that in control group (P<0.05).In addition,the ratio of Treg/Th17 cells was lower in model mice than that in control group.Conclusion: The imbalance of CD4+CD25 Treg/Th17 cells may participate in the pathogenesis of EAPS.

Objective To construct an eukaryotic expression vector for shRNA targeting WT1 gene.Methods Firstly, one site targeting human WT1 gene in gene bank was selected according to the guidelines for the selection of highly effective siRNA sequence and then BLAST test was performed. Secondly,a pair of oligonucleotides fragments were synthesized and annealed to double strand. Thirdly, the double strand was cloned into plasmid vector pGenesil-1. Finally, the recombinant plasmid was identified by restriction enzyme digestion and sequencing analysis. Results Enzyme cutting, and sequencing analysis have confirmed that the construction was successful. Conclusion WT1shRNA is constructed which is the fundament to further research of WT1 gene.

BACKGROUND: Classic isolation method of bone marrow mesenchymal stem cells (BMSCs) is Percoll density gradient centrifugation. Blood cell component was removed. However, this method is complicated. Preparation density was needed when isolating dog bone marrow. Moreover, centrifugation was frequent, which had a great damage to cells. OBJECTIVE: To establish methods of the isolation, proliferation, culture and osteoinduction of canine BMSCs, and observe the in vitro proliferation and ability to osteoinduction differentiation. METHODS: 10 mL bone marrow was extracted from dog posterior superior iliac spine, heparin anticoagulation, diluted using Hanks juice, treated with 1.077 g/mL Ficoll solution 3 mL, and centrifuged at 2 000 r/min for 20 minutes. Karyocytes were absorbed to form white cloudlike layering interface, and then centrifuged twice using DMEM supplemented with fetal bovine serum, incubated at 12×10~4/cm~2 at 37 ℃ in a 5% CO_2 incubator. Following subculture, cells were incubated in DMEM containing dexamethasone, β-sodium phosphoglycerol and ascorbic acid 2-phosphate. Immunocytochemical staining and immunofluorescence staining were utilized to detect osteocalcin, osteopontin and type Ⅰ collagen expression in osteoblasts. Alkaline phosphatase staining and alizarin red staining were performed. RESULTS AND CONCLUSION: 1.077 g/mL Ficoll density gradient centrifugation was used to isolate karyocytes that were significant compared with Percoll solution. Obtained BMSCs had high purity, good growth and the mean doubling time was 24 hours. Following in vitro osteogenic incubation of dog BMSCs, osteocalcin, osteopontin and type Ⅰ collagen showed positive expression. Alkaline phosphatase staining demonstrated bluish-green cytoplasm. Alizarin red staining showed red nodes in extracellular matrix, and could differentiate into osteoblasts in vitro.

Objective To construct a fusion gene (h1a-spaO) encoding H1a-SpaO protein of Sal-monella paratyphi A ( S.paratyphi A) and to express it in prokaryotic expression system , then to further ana-lyze the immunoprotective effects of the expressed protein rH 1a-SpaO.Methods The h1a-spaO fusion gene formed from separate h1a and spaO genes was amplified by PCR using flexible peptide sequence-containing linking primers and then sequenced after T-A cloning.A prokaryotic expression system for expressing h1a-spaO fusion gene was constructed by using the genetic engineering technique .The expressed protein rH1a-SpaO was examined by SDS-PAGE.The antigenicity and immunoreactivity of rH1a-SpaO protein were deter-mined by Western blot assay .The ability of rH1a-SpaO antiserum agglutinating S.paratyphi A strains was detected by micro-Widal′s test.The immunoprotective effects of rH 1a-SpaO against the lethal dose challenge of S.paratyphi A strains were analyzed in a mouse model and that were compared with those by using equal dose of individual recombinant protein H1a and SpaO (rH1a and rSpaO) as the immunogens, respectively. Results The h1a-spaO fusion gene was 100%identical with the individual h1a or spaO gene in nucleotide and amino acid sequences .The constructed prokaryotic expression system could express the recombinant pro-tein rH1a-SpaO with an advantage of high efficiency .rH1a-SpaO protein was able to react with rH 1a or rSpaO antiserum.Moreover, rH1a-SpaO antiserum also could efficiently recognize rH 1a and rSpaO as well as agglutinate Salmonella paratyphi A strains by binding with H-antigen.The immunoprotective rate (93.3%) in mice pre-immunized with 100 μg of rH1a-SpaO protein was significantly higher than that in those pre-immunized with equal dose of rH1a (60.0%) protein or rSpaO protein(53.3%) (P<0.05).Conclusion The recombinant fusion protein rH 1a-SpaO showed more stronger immunoprotective function than the individ-ual rH1a or rSpaO protein , which could be used as an effective antigen for the development of bi -valent para-typhoid A vaccine or typhoid/paratyphoid capsular polysaccharide-protein combined vaccine .

Objective To investigate the effect to the expression of VEGF in human leukemic cell line by WT1gene silencing.MethodsK562 cells were transfected with WT1shRNA Plasmid.Transfection efficiency was detected by flow cytometIy,and the succeed transfered cells Were sorted.WT1 and VEGF mRNA variation were measured by fluorescence RT-PCR.VEGF concentrations in the cell growth medium were detected by ELISA.Results 48 hours after transfection,the transfection efficiency was 65％±4.5％.WT1shRNA positive cells were sorted by flow cytometry.Both the mRNA level of WT1 and VEGF and VEGF concentrations in cell growth medium were significantly decreased than the control group.ConclusionWT1 gene silence can down-regulate the expression of VEGF in K562 cells which suggest that WT1 gene may take part in leukemia angiogenesis through up-regulation of VEGF.

Objective To study the subacute inhalation toxicity of Ivermectin TC, and obtain its non-observed adverse effect level(NOAEL).Methods It was performed on the doses of Ivermectin TC 190, 380, 750 mg/m3, the solvent control group (0.03%Tween-80 solution) , the control group and additional group ( there were 6 female and 6 male Sprague-Dawley rats for each group) .The animals inhaled with nose-only exposure for 4 weeks (4 h/d, 5 d/week) .The additional group should be observed another 14 days after exposing.At the end of experiment, the rats were killed, the routine and biochemical detection, the body weight and organ to body weight ratios were all measured.Results In the high exposure group, clinical signs of rats included hair fluffy, dull, salivation, tremors were recorded at the exposure period;in female rats, feed efficiency was decreased, ALT and liver to body weight ratio were increased; in male rats, BUN and ALT were increased, CHOL and body weight for the 4th week were decreased.Histopathological examinations revealed that swelling in the liver cell was seen in some female rats at high exposure group.Conclusion The results suggested that the NOAEL of Ivermectin TC in SD rats was 380 mg/m3(4 h/d for 28 days).

Objective:To research rmIL-18 in vitro culture system CCs induce tumor-specific cytotoxic T lymphocytes CTL and anti-tumor effect in mice.Methods:Used Stem SepTM immune magnetic cells separation method to culture mouse spleen NK cells,T cells and DCs,established culture systems in vitro;used of different approaches,different doses rmIL-18 to immunize HCC tumor-bearing mice,researched the effect of rmIL-18 on tumor growth rate and survival time.Results:rmIL-18 could induce and promote tumor-specific CTL-mediated killing effects in vitro culture system;tumor-specific CTL could significantly inhibit tumor growth(P<0.01) of and prolong the survival time of liver cancer tumor-bearing mice(P<0.01),and the effect was increased with rmIL-18 concentration increased(P<0.01),and intratumoral injection was superior to intraperitoneal injection(P<0.01).Conclusion:rmIL-18 can induce tumor-specific CTL in vitro and play a role in anti-liver cancer in mice.