Animals ; Beverages ; Macrophages ; drug effects ; metabolism ; Mice ; Mice, Inbred Strains ; Receptors, Complement 3b ; metabolism

AIM:To investigate the clinical effect of anti-vascular endothelial growth factor (VEGF) drugs combined with laser photocoagulation for diabetic macular edema (DME).METHODS: Totally 94 patients (141 eyes) with DME from June to December 2015 in our hospital were selected and randomly divided into combined group of 47 cases (68 eyes, ranibizumab combined with laser therapy) and the control group of 47 cases (73 eyes, laser treatment).The levels of best corrected visual acuity (BCVA), macular central retinal thickness (CRT), total macular volume (TMV) and macular edema level were compared between the two groups at different time after treatment.RESULTS: The mean values of BCVA in the combined group were higher than those in the control group at 2, 6 and 12wk, and the difference was statistically significant (P<0.05).At 2, 6 and 12wk after treatment, the CRT and TMV values of the combined group were lower than those of the control group, the difference was statistically significant (P<0.05).After treated for 12wk, patients with macular edema of combined group was 80.9% in mild level, 17.7% in moderate level, 1.5% in severe level, those of the control group was 60.0%, 31.5%, 5.5%, the difference between the two groups was statistically significant (P<0.05).CONCLUSION: The effect of anti-VEGF drugs combined with laser therapy for DME patients is better than that of single laser therapy alone.

Human ribosomal DNA (hrDNA) targeting vector pHr is a homologous recombinant plasmid for human genome which developed in the State Key Laboratory of Medical Genetics. pHr was used to construct a recombinant plasmid pHr-CMG expressing mda-7/GFP fusion gene and its efficacy in the hepatocellular carcinoma cell line Bel-7402 was investigated. The expression of mda-7/GFP fusion gene was detected by fluorescent microscope, RT-PCR and Western blotting, and its function was detected by cell-cycle analyses, MTT assay and Hoechst33258 staining. The results demonstrated that pHr-CMG vector could express MDA-7/GFP fusion protein effectually and the mda-7 gene could induce cell apoptosis and proliferation suppression in Bel-7402 cell line, which might be caused by the G2/M cell cycle arrest. These results also suggested that human ribosomal DNA targeting vector system and the pHr-CMG vector may be applied in further gene therapy researches for hepatocellular carcinoma.

In order to study the tumor suppression effect of p53 with CMV enhancer and hTERT promoter mediated by human-derived vector pHrn in liver cancer cell Bel-7402, report plasmid pchEGFP, tumor suppressor plasmids pchp53Arg and pchp53Pro were constructed by inserting expression cassette CMVe+hTERTp+EGFP, CMVe+hTERTp+p53Arg and CMVe+hTERTp+p53Pro into pHrn respectively. 24 h after cell transfection by lipofectamine 2000, GFP expression pattern was analyzed through fluorescence microscope and flow cytometry; RT-PCR and Western blot were taken to study the p53 expression pattern. The cell apoptosis by Hoechst 33258 and Annexin V-FITC/PI staining was also studied. Results show that the expression of GFP and p53 protein in Bel-7402were detected, but apparent cell apoptosis could not be found. The recombinant p53 mediated by human-derived vector could express in Bel-7402, but no significant tumor suppression effect was detected, which might result from the down regulation effect of the wild type p53 on hTERT promoter.

Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Gene Library ; Hepatitis B Surface Antigens ; immunology ; metabolism ; Hepatitis B virus ; genetics ; immunology ; Humans ; Immunoprecipitation ; Intracellular Signaling Peptides and Proteins ; metabolism ; Nerve Tissue Proteins ; metabolism ; Viral Proteins ; immunology ; metabolism

Objective To investigate the effect of continual care on postoperative medication compliance of patients with osteoporotic vertebral compression fractures (OVCF) after vertebroplasty.Methods A retrospective case control study was performed to analyze the clinical data of 150 patients with OVCF who underwent vertebroplasty in Guizhou People's Hospital from January 2016 to May 2017.There were 38 males and 112 females,aged 47-88 years,with an average of 67.5 years.Seventy-five patients were given continual care such as telephone follow-up and home visit (continual care group).Seventy-five patients were given routine health education (routine care group) when they were discharged from hospital.Visual analogue scale (VAS) and Oswestry dysfunction index (ODI) before operation,1 day,1,3,6 and 12 months after operation,as well as medication compliance and vertebral re-fracture at 1,3,6 and 12 months after operation were compared between the two groups.Results The preoperative VAS of the routine care group was 6 (6-7),1 (0-1),1 (1-3),2 (1-3) and 3 (2-5) points at 1 day and 1,3,6 and 12 months after operation,respectively.The preoperative ODI was 21 (18-27),0 (0-0),2 (1-4),5 (3-7),7 (5-10),10 (7-14) points at 1 day,1,3,6 and 12 months after operation.In the continual care group,VAS was 7 (6-7) points before operation,0 (0-1),1 (0-1),1 (0-2) and 2 (1-3) points at 1 day,1,3,6 and 12 months after operation,respectively.ODI of the continual care group was 18 (22-28) points before operation,0 (0-1),2 (0-4),4 (1-5),4 (3-6) and 6 (4-9) at 1 day,1,3,6 and 12 months after operation.The VAS and ODI of the two groups were lower than those before operation,and the scores of the continual care group were lower than those of the routine care group at 1,3,6 and 12 months after operation (P ＜ 0.05).The medication compliance rates of continual care group were 93％,89％,91％ and 84％ at 1,3,6 and 12 months after operation,while those of routine care group were 44％,40％,47％ and 40％ respectively (P ＜0.05).The incidence of vertebral re-fracture was 1％,1％,3％ and 3％ in continual care group and 3％,5％,5％ and 7％ in routine care group at 1,3,6 and 12 months after operation respectively (P ＜ 0.05).Conclusion Continual care can improve the medication compliance of OVCF patients after treatment with vertebroplasty,relieve pain,improve the quality of life,and reduce the incidence of vertebral re-fracture,which is worthy of clinical promotion.

Objective To explore the chemotherapeutic effect of gemcitabine for non-small cell lung cancer patients and analyzed the correlation of chemotherapeutic effect with ribonucleotide reductase M1 (RRM1),excision repair cross-complementing 1 (ERCC1) expression in pathological tissue.Methods The clinical data of 96 non-small cell lung cancer patients were selected.All patients were treated with gemcitabine + cisplatin chemotherapy,and the RRM1,ERCC1 expression in pathological tissue were detected by immunohistochemical method at 3 weeks after chemotherapy,the correlation of clinical features with chemotherapeutic effect were analyzed.Results The positive expression rates of RRM1 and ERCC1 were 60.42％,43.75％.The positive expression of non-small cell lung cancer patients pathological tissue RRM1,ERCC1 had significant correlation with the gender and smoking status (P ＜0.05 or P ＜0.01),while had no correlation with the age and clinical stages (all P ＞0.05).The effective rates of patients with pathological tissue RRM1/ERCC1 +/+,-/-,+/-or-/+ were 17.95％,74.29％ and 59.09％,and the effective rates of patients with pathological tissue RRM1/ERCC1-/-were higher than +/ +,+/-or-/+ (P ＜0.05 or P ＜ 0.01),and the effective rates of patients with pathological tissue RRM1/ERCC1 + /-or-/ + were higher than RRM1/ERCC1 + / + (all P ＜ 0.05).Spearman correlation analysis showed that,the negative expression of non-small cell lung cancer patients anthological tissue RRM1,ERCC1 had significant positive correlation with chemotherapeutic effect (r =0.248,0.306,all P ＜ 0.05).Conclusion Gemcitabine has a certain clinical effect in the treatment of non-small cell lung cancer patients,and the clinical effect had significant positive correlation with the negative expression of non-small cell lung cancer patients anthological tissue RRMl,ERCC1 protein,which can act as the evaluation index for the chemotherapeutic effect.

OBJECTIVETo analyse the male reproductive toxicity of carbaryl.

METHODSThirty-one male carbaryl exposure workers and 46 male administrators in the office in a pesticide factory were selected as the exposure group and internal control group respectively, and 22 male administrators in a center for disease control and prevention were served as the external control group. In order to evaluate the exposure levels, the concentrations of carbaryl, methyl isocyanate, ammonia and phenol in the ambient air of the work place in these three groups were monitored simultaneously for three consecutive days. Moreover, three workers in the exposure group and the external control group were selected to evaluate the amount of carbaryl of individual and dermal contamination for three consecutive days. After the semen were collected according to the standard method, the workers'semen qualities were analysed with WHO method, the sperm morphology and the sperm motility were evaluated using micro-cell slide spectrum technology and computer assisted sperm analysis (CASA) respectively.

RESULTSIn the exposure group, the concentrations of carbaryl and phenol (52.41 mg/m(3) and 0.08 mg/m(3) respectively) were significantly higher than those in the internal and external control group (P < 0.01 or P < 0.05). Furthermore, in the carbaryl exposure area the geometric mean concentration of carbaryl with the individual sampling was 7.38 mg/m(3), and the geometric mean of dermal contamination detected in the carbaryl exposure area was 862.47 mg/m(2). Carbaryl was not found in the external control area (P < 0.01). The seminal volume [(2.39 +/- 1.44) ml] and the sperm motility [(1.77 +/- 0.61) grade] were significantly lower than those in the external control group (P < 0.05), and sperm motion parameters such as linearity (LIN, 39.89% +/- 6.00%), straightness (STR, 71.51% +/- 11.22%), straight line velocity [VSL, (26.29 +/- 7.84) microm/s] and beat cross frequency [BCF, (3.99 +/- 1.55) Hz] were lower than those in the internal and external control group (P < 0.05), while the abnormal rates of viscidity, sperm motility and total aberration rate were higher than those in the external control group (P < 0.05).

CONCLUSIONOccupational exposure to carbaryl production can affect the workers'sperm and semen quality to certain extent.

Adult ; Carbaryl ; adverse effects ; Humans ; Insecticides ; adverse effects ; Male ; Occupational Exposure ; Semen ; drug effects ; Sperm Motility ; drug effects ; Spermatozoa ; abnormalities ; drug effects

OBJECTIVETo construct a human source vector containing minidystrophin-EGFP fusion gene and investigate its expression in Cos-7 cells.

METHODSThe recombinant human source vector named pHrnDysG was constructed with PCR-clone methods. Three fragments of dystrophin gene were PCR amplified from normal human dystrophin gene cDNA (GenBank NM04006). These three fragments were ligated to generate a minidystrophin gene. The enhanced green fluorescent protein (EGFP) gene was fused to the C terminal of the minidystrophin gene, and then the pHrnDysG was finally obtained by cloning the fusion gene to pHrneo. Fluorescence microscope and RT-PCR were used to detect the expression of minidystrophin-EGFP fusion gene after the recombinant construct was transfected into Cos-7 cells by lipofectamine.

RESULTSRestrictive enzyme digestion analysis and sequencing confirmed that pHrnDysG vector was constructed successfully. After the recombinant pHrnDysG was transfected to Cos-7 cells, RT-PCR demonstrated that the fusion gene was successfully transcribed, and the green fluorescence was observed at the cell membrane.

CONCLUSIONThe minidystrophin-EGFP fusion gene mediated by pHrneo vector could express in Cos-7 cells and its products' localization in the cell membrane was the same as that of full length dystrophin. These results suggested that the recombinant human source vector pHrnDysG might be potentially used in studies on the gene therapy of Duchenne muscular dystrophy.

Animals ; COS Cells ; Cercopithecus aethiops ; Dystrophin ; genetics ; metabolism ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Models, Genetic ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVE: To investigate the biological characteristics of endothelial progenitor cells (EPCs) from the umbilical cord blood (UCB), and to evaluate their oncogenicity after long-term culture in vitro. METHODS: The mononuclear cells (MNCs) were isolated from the UCB and cultured in MCDB131 medium supplemented with 20% FBS, VEGF and other growth factors. Morphology of the EPCs was observed, and the growth curve of the EPCs was investigated. Surface antigens of the EPCs were analyzed by the flow-cytometer. The capability of intaking the acetylated low-density lipoprotein (acLDL) of the EPCs was detected using fluoresencent chemical method. The vasoformative capability and genetic stability of EPCs were cultured in matrigel, and examined by karyotype analysis. The oncogenicity of EPCs was verified by the tumorigenesis test in athymic mouse and soft agar. RESULTS: EPCs were successfully derived from the UCB, and could be passaged to at least 42(nd) generation and had strong abilities of proliferation, acLDL intake and vasoformation, but there was not oncogenicity. They expressed endothelial cell-surface antigens and maintained normal karyotype. CONCLUSION The EPCs with proliferative potential can be isolated from the UCB. They can be passaged in long-term cultures without oncogenicity, and can maintain normal karyotype. The EPCs can be served as a new type of cells in cell and gene therapy.
Animals ; Antigens, Surface ; analysis ; Cell Line ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Fetal Blood ; cytology ; Flow Cytometry ; HeLa Cells ; Humans ; Infant, Newborn ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Karyotyping ; Mice ; Mice, Nude ; Neoplasms, Experimental ; pathology ; Stem Cells ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; pharmacology