Background Aberrant proliferation of residual lens epithelial cells (LECs) is one of main causes of posterior capsular opacification (PCO).Researches indicated that Wnt3a signaling pathway promote proliferation of epithelial cells,but its effect on LECs is still unclear. Objective The present study was to investigate the effects of Wnt3a on proliferation of human LECs and its mechanism and to provide a new gene target in the prevention and treatment of PCO. Methods Human LECs line (SRA01/04 cells ) was cultured and then incubated to 6-well plate at the density of 4×105/well.A human Wnt3a cDNA expressing vector targeted human LECs was constructed to increase the Wnt3a expression in SRA01/04 cells,and pcDNA3-HA expression vector was used as the control group.The expression of Wnt3a was identified by Western blot assay after transfected.The growth and proliferation of SRA01/04 cells were detected by MTT and flow cytometry (FCM).The expressions of β-catenin,cyclin D1 and c-myc in the cells were detected by Western blot assay.β-Catenin expression was localized using immunofluorescence assay,and the expression and localization of proliferating cell nuclear antigen ( PCNA ) were analyzed by immunocytochemistry for the exploration of the active mechanism of Wnt3a to proliferation of LECs.Results Human Wnt3a cDNA expression vector was designed successfully and transiently transfected to SRA01/04 cells,and Wnt3a/SRA01/04 cells and pcDNA3-HA/SRA01/04 cells were obtained.The expression of Wnt3a was verified in the Wnt3a transfected group compared with the control group.MTT indicated that the cell proliferating rate was significantly different between the Wnt3a transfected group and the control group ( Fgroup =15.235,P =0.005 ;Ftime =369.677,P =0.000),and that in various time points after transfected was significantly different (t =20.843,P=0.001 ;t =26.214,P＜0.001 ;t=25.177,P=0.001 ;t =35.516,P＜0.001 ;t =615.056,P＜0.001 ).The proportion of SRA01/04 cells in G1 phase was 51.74％ in the Wnt3a cDNA transfected group,with a significantly decrease in comparison with 79.44％ of the control group.However,the proportion of SRA01/04 cells in S phase in the Wnt3a cDNA transfected group was higher than that of the control group (36.23％ versus 12.34％ ).The positive expression rate of PCNA protein in SRA01/04 was (47.00％ ±7.58％ ) in the Wnt3a cDNA transfected group and ( 16.00％ ±3.61％ ) in the control group with a significant difference between them (t =8.256,P＜0.01 ).After 48 hours of transfection of the Wnt3a cDNA,the expression amount of β-catenin proteins was higher and the immunofluorescence was stronger in cell nucleus,and the expressions of cyclin D1 and c-myc proteins were elevated in Wnt3a/SRA01/04 cells. Conclusions The overexpression of Wnt3a activates the Wnt/β-catenin signaling pathway and downregulates the expression of a subset of target genes,including cyclin D1 and c-myc,which plays an important role in promoting the proliferation of human LECs.

Dentists ; Disasters ; Earthquakes ; Humans ; Oral Surgical Procedures

OBJECTIVETo investigate the effect of hypoxia on the human pulmonary artery smooth muscle cells two pore domain potassium channels TASK-1 and the regulation of non-receptor tyrosine kinase c-Src in this process.

METHODSThe cultured human pulmonary artery smooth muscle cells (hPASMCs) were divided into: normal group, hypoxia 30 minute group, hypoxia 6 hours group and hypoxia 48 hour group, and hypoxia 48 hour + PP2 group, hypoxia 48 hour + PP3 group, hypoxia 48 hour + bpV group. Flow cytometry was used to analyze the cell cycle, RT-PCR and Western blot technique were carried out to detect the expression changes of TASK-1 mRNA and protein in different groups.

RESULTS(1) Cell Cycle Show: Compared with normal control group, with prolonged hypoxia, the percentages of hPASMCs in S phases of cell cycle were increased. While compared with hypoxia 48 hour group, the percentages of hypoxia 48 hour + PP2 group hPASMCs in S phases of cell cycle were decreased. The expression of TASK-1 mRNA on hPASMCs in acute hypoxia 6 hour group was increased, while the expression of TASK-1 protein on hPASMCs in the acute and chronic hypoxia group was decreased, and the expression of TASK-1 mRNA on hPASMCs in the chronic hypoxia group was decreased; After pre-incubation of a potent and selective inhibitor of the Src family of protein tyrosine kinases PP2, the expression of TASK-1 mRNA and protein in hypoxia 48 hour group was increased, however after pre-incubation of the inhibitor of the Src family of protein tyrosine phosphatase bpV, the expression of TASK-1 protein in hypoxia 48 hour group was decreased.

CONCLUSIONHypoxia promotes human pulmonary artery smooth muscle cell proliferation, and non-receptor tyrosine kinase c-Src may participate in the expression of two pore domain potassium channels TASK-1 regulated by hypoxia. Therefore, we hypothesized that TASK-1 channels and c-Src participatein the acute and chronic hypoxic human pulmonary vasoconstriction.

Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Humans ; Myocytes, Smooth Muscle ; cytology ; Nerve Tissue Proteins ; metabolism ; Potassium Channels, Tandem Pore Domain ; metabolism ; Pulmonary Artery ; cytology ; RNA, Messenger ; Vasoconstriction ; src-Family Kinases ; metabolism

Objective: To observe the clinical efficacy of heat-sensitive moxibustion plus Western medicine in treating patients with diabetic peripheral neuropathy (DPN). Methods: A total of 70 patients with DPN were divided into an observation group and a control group by sealed envelope method combined with the random number table method, with 35 cases in each group. The control group was treated with routine medicine, and the observation group was treated with heat-sensitive moxibustion on the basis of the treatment in the control group. After 2 courses of treatment, the scores of Toronto clinical scoring system (TCSS) and vibration perception threshold (VPT) in both groups were observed, and the clinical efficacy was compared. Results: During treatment, 3 cases dropped out in the control group and 4 cases in the observation group. After treatment, the total effective rate in the observation group was higher than that in the control group (P<0.05). The scores of TCSS and VPT in both groups decreased after treatment, and the intra-group comparison showed statistical significance (both P<0.05). The scores of TCSS and VPT in the observation group were lower than those in the control group, and the differences were statistically significant (both P<0.05). Conclusion: Heat-sensitive moxibustion plus Western medicine can improve the symptoms in patients with DPN, and has a better curative effect than the Western medicine alone.

Background The surgery for femtosecond laser created laser in situ keratomileusis(LASIK)flaps has made great progression recent year,but the postoperative corneal wound healing and regeneration of nerve fibers after surgery are closely concerned.Objective This study was to compare and analyze the clinical outcomes between FEMTO LDV femtosecond laser flap and mechanical microkeratome Hansatome flap in LASIK.Methods A prospective case-controlled study was designed.The serial 38 myopic eyes of 38 patients were included from March through July,2010 in Henan Armed Police Force General Hospital.The patients were randomized into FEMTO LDV femtosecond laser assisted flap group(20 patients/20 eyes)and mechanical microkeratome Hansatome assisted flap group(18 patients/18 eyes)with the matched age,gender and refraction of spherical equivalent.HRT Ⅲ examinations were performed before surgery,1 week,1 month and 3 months after surgery to compare the morphological changes atthe center and margin of the flaps,and evaluate the similarities and differences of cellular morphology after surgery between the two approaches.Written informed consent was obtained from each patient prior to this medical trial.Results The best corrected visual acuity was ≥ 1.0 and the refract diopter was similar in both groups(+0.21 D±0.48 D and-0.04 D±0.54 D)1 month after LASIK.The corneal thickness was insignificant increased in the first week after LASIK,and the density of shallow stromal cells was decreased in 1 week,1 month and 3 months compared with pre-operation in the femtosecond laser assisted flap group(t =-27.99,-25.49,-28.87,P ＜ 0.01).In the Hansatome assisted flap group,significantly thickened corneal epithelium was seen in the first week after LASIK compared before LASIK(56.73 μm±2.47 μm versus 51.16 μm±1.11 μm)(t=9.29,P＜0.05),and the density of shallow stromal cells was decreased in 1 week,1 month and 3 months compared with pre-operation in the Hansatome assisted flap group(t =-17.57,-14.13,-19.63,P =0.00).The density of high reflective interface particles in cornea was lower in 1 week,1 month and 3 months after LASIK in the femtosecond laser assisted flap group than that in the Hansatome assisted flap group,showing significant differences between them(t =-13.505,-11.900,-14.084,P＜0.01).The active stromal cells were seen beneath the interface in both groups in the first week and gradually decreased after that time.Intact corneal nerve fibers were found in the femtosecond laser assisted flap group,but those in the Hansatome assisted flap group were shorter and smaller 3 months after LASIK.At 3 months after surgery,the flap margin showed stromal higher reflection and irregular secondary fibrosis in the femtosecond laser assisted flap group,and in contrast,the flap margin had the appearance of a unclearly identified fibrotic scar in the Hansatome assisted flap group.Conclusions Compared with the LASIK and Hansatome assisted flap,the LASIK with FEMTO LDV flap shows earlier nerve fiber regeneration and greater fibrotic scarring,which imply a good wound healing process in the LASIK with FEMTO LDV flap.

The high-copy-number plasmid pcDNA3.1+ is unstable within S almonella typhimurium. A novel plasmid pmcDNA3.1+ was constructed by removin g the promoter sequence of ampicillin resistance gene (bla gene) in plasmid pcDNA3.1+. In contrast to pcDNA3.1+, pmcDNA3.1+ was stable within Salmonel la typhimurium SL7207 in LB medium with or without ampicillin. Further experi ments showed the ?-lactamase activity of Salmonella typhimurium SL7207(pmc DNA3.1+) was apparently lowered than that of Salmonella typhimurium SL7207( pcDNA3.1+) and the high ampicillin concentration was maintained longer in LB me dium culturing Salmonella typhimurium SL7207(pmcDNA3.1+). When mice were a dministered with Salmonella typhimurium SL7207(pmcDNA3.1+) intraperitoneall y, more than 95% of Salmonella cells separated from the spleen still harbore d the plasmid pmcDNA3.1+ 7 days later; but 99% of Salmonella cells lost the plasmid pcDNA3.1+ at day 3 in mice innoculated with Salmonella typhimurium SL7207(pcDNA3.1+). By lowering the expression of bla gene, the rapid deco mposition of ampicillin in LB medium was avoided and the metabolic pressure was relieved for the host cells. This method offers a solution for the problem of t he instability of high-copy-number plasmid within Salmonella typhimurium.

OBJECTIVETo establish a stable H460 cell line with Nrf2 down-regulation to study the role of Nrf2 in Oxaliplatin resistance.

METHODSNrf2 down-regulated H460 cell line was obtained by transfecting cells with pRS-hNrf2 and followed by screening for Nrf2 expression by Western blotting. The cell lines were further characterized by analysing cellular GSH levels and cytotoxicity to anti-cancer drugs with MTS.

RESULTNrf2 down-regulated H460 cell lines were established successfully. Cellular GSH level reduced significantly in the H460-C9 and H460-C13 compared to control cells (P(C9)= 0.00249, P(C13)= 0.03944). Down-regulation of Nrf2 in H460 sensitized the anti-cancer effect of Oxaliplatin and Doxorubicin.

CONCLUSIONNrf2 plays an important role in drug resistance. Down regulation of Nrf2 in H460 cell enhances cytotoxicity of Oxaliplatin.

Antineoplastic Agents ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Drug Resistance, Neoplasm ; genetics ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Organoplatinum Compounds ; pharmacology ; Transfection

Cancer, an abnormal cell proliferation resulted from multi-factors,has the highest morbidity and mortality among all the serious diseases. Considerable progress has been made in cancer biology in recent years. Tumor immunology, cancer stem cells (CSCs), autophagy, and epithelial-mesenchymal transition (EMT) have become hot topics of interests in this area. Detailed dissection of these biological processes will provide novel directions, targets, and strategies for the pharmacological evaluation, mechanism elucidation, and new drug development of traditional Chinese medicine.
Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Humans ; Neoplasms ; drug therapy ; genetics ; immunology ; physiopathology

To examine the phylogenetic information regarding the gene pool of AIV in domestic ducks in eastern China, the NA genes of twenty-six viruses isolated during 2002-2006, including two H1N1 strains, tenH3N1 strains and fourteen HSN1 strains, which reflected the predominant N1 subtype viruses were subjected to phylogenetic analysis. The results indicated that AIVs of N1 subtype circulating in domestic ducks in eastern China were undergoing a gradual evolution. Analysis of the deduced amino acid sequences revealed that NAs from all isolated H5N1 viruses had a 20-aa deletion in the stalk region (residues 49-68), whereas no deletion was seen in the NAs from other HA subtype viruses. The viruses of H3N1 and H1N1 might have a propensity for reassortment of NA genes, whereas no direct evidence of reassortment of NA gene was obtained in H5N1 viruses.
Animals ; Birds ; China ; DNA, Viral ; analysis ; Evolution, Molecular ; Humans ; Influenza A Virus, H1N1 Subtype ; classification ; genetics ; Influenza A Virus, H3N2 Subtype ; classification ; genetics ; Influenza A Virus, H5N1 Subtype ; classification ; enzymology ; genetics ; Influenza A virus ; classification ; enzymology ; genetics ; Influenza in Birds ; virology ; Influenza, Human ; virology ; Neuraminidase ; genetics ; Phylogeny ; Poultry Diseases ; virology ; Sequence Alignment ; Sequence Deletion

OBJECTIVETo study the pathological change of rats' benign hyperplastic prostate (BHP) after radical denervation.

METHODSA total of 65 male spontaneous hypertension rats (SHR) at 30 weeks age were randomly assigned into treatment group, sham surgery control group and normal control group. In surgery group, all the axonal branches of the major pelvic ganglion (MPG) supplying the bilateral prostate were truncated, followed performing of cystostomy; In sham surgery control group, only cystostomy was performed; In normal control group, no procedure was performed. The rats were sacrificed at 3, 7, 11, 15 and >or= 21 d post-operation respectively. The gross morphological changes of prostate in all animals were observed.

RESULTSIn treatment group, the prostate in 3 d post-operation showed granular solidification and shrunken volume and the changes occurred gradually over time. The glandular epithelial cells showed gradual degeneration, necrosis and detachment. The glandular epithelium became progressively thinner, the smooth muscles elongated and thinned progressively and the stromal components showed mild to moderate overgrowth. At the later stage, the glandular epithelium, glandular lumen and smooth muscles gradually disappeared and the prostate was largely replaced by connective tissues. Electron microscopic study showed that the glandular cells gradually underwent vacuolar degeneration and the structures of basement membrane became fuzzy. The smooth muscles cells degenerated overtime and the fibroblasts and collagenous fibers in the stroma overgrew slowly. At the late stage, most of the glandular cells became necrotic, the basal membrane and smooth muscle cells disappeared and collagenous fibers were highly hyperplasic. In surgery group in 3 d post-operation, the S-100 staining of nerve fiber was diffuse and disappeared after 11 d while it persisted normally in other groups. The two values in sham surgery control group showed no significant changes post-operatively.

CONCLUSIONSAfter radical denervation, the rat prostate with benign hyperplasia (gland and smooth muscles) undergoes dramatic atrophic changes and the volume decreases significantly. It suggests that this treatment may represent a novel therapy for BPH.

Animals ; Denervation ; methods ; Disease Models, Animal ; Male ; Microscopy, Electron, Transmission ; Prostate ; innervation ; pathology ; ultrastructure ; Prostatic Hyperplasia ; pathology ; surgery ; Random Allocation ; Rats ; Rats, Inbred SHR