Air Pollutants, Occupational ; analysis ; Chlorofluorocarbons, Methane ; analysis ; Chromatography, Gas ; methods ; Workplace

ObjectiveTo investigate the effects of ABI-1 gene knockdown upon the proliferation and migration of human gastric cancer cell NCI-N87 in vitro. MethodsNCI-N87-ABI-I-ShRNA cell model was successfully constructed and validated by Real-time PCR and Western blot. The cellular morphous and skeleton, proliferative and migrative potents, and also AKT expression were compared between NCI-N87-ABI-1-ShRNA and its parents by immunofluorental staining, CCK-8 assay, transwell chamber and Western blotting.ResultsCCK-8 assay showed there was no significant difference in the proliferation rates at different time points between the NCI-N87-Vector and NCI-N87 cells while the proliferation rates at the time points of 36 and 48 hours of the NCI-N87-ABI-1-ShRNA were significantly lower than the NCI-N87( t =2. 85and 4. 166, P ＜ 0. 05 ). Transwell assay showed that migrated cell number were 66 ± 8, 65 ± 8 and 30 ± 4,respectively, and there was significant difference between the NCI-N87-ABI-1-ShRNA and NCI-N87 cells (t =9. 550,P ＜0. 05). Finally, ABI-1- knock-down altered the cellular morphoos and skeleton of 90％NCI-N87 cells and inhibited p-AKT expression.ConclusionABI-1 inhibits proliferation and migration of NCI-N87 cells in vitro probably by PI3K/AKT pathway.

In order to develop a combined live vaccine that will be used to prevent against porcine parvovirus (PPV) and Pseudorabies virus (PRV) infection, the VP2 gene of PPV was inserted into the transfer vector plasmid pG to produce the recombinant plasmid pGVP2. The plasmid pGVP2 and the genome of PRV HB98 attenuated vaccine were transfected by using lipofectamine into swine testis cells for the homologous recombination. The recombinant virus rPRV-VP2 was purified by selection of green fluorescence plaques for five cycles. 6-week-old female Kunming mice were immunized intramuscularly with attenuated PRV parent HB98 strain, commercial inactivated vaccine against PPV, recombinant virus, DMEM culture solution. The injections were repeated with an equivalent dose after 2 weeks in all of the groups, and then challenged with the virulent PRV NY strain at 7 weeks after the first immunization. The recombinant virus rPRV-VP2 was successfully generated, and the recombinant virus could effectively elicite anti-PPV and PRV antibody and significant cellular immune response as indicated by anti-PPV ELISA and HI, PRV-neutralizing assay and flow cytometry. The challenge assay indicated that recombinant virus could protect the mice against the virulent PRV challenge. These results demonstrated that the recombinant virus can be a candidate recombinant vaccine strain for the prevention of PRV and PPV.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; administration & dosage ; genetics ; immunology ; Capsid Proteins ; administration & dosage ; genetics ; immunology ; Female ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Herpesvirus 1, Suid ; genetics ; metabolism ; Mice ; Parvovirus, Porcine ; genetics ; immunology ; Swine ; Swine Diseases ; immunology ; prevention & control ; virology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

Background The condition of the sclera is associated with many ocular diseases.The measurement of human scleral thickness in vivo is helpful for us to understand the features of the sclera and related diseases.Objective The present study was to measure the anterior sclera thickness(AST) in patients with senile cataract and to analyze the relationship among AST and other associated ocular parameters.Methods This study was approved by the Ethic Commission of First Hospital of Peking University.Written informed consent was obtained from each individual prior to examination.One hundred and five senile cataract patients were recruited in this study.Central corneal thickness (CCT),corneal curvature (CCV) and axial length were measured using ultrasonic pachymeter,keratometer,and A-scan unit,respectively.The AST was measured at 2 mm posterior to the scleral spur in the temporal meridian using ultrasound biomicroscope (UBM).The differences of CCT,CCV,ocular axial length and AST between bilateral eyes and the different sexes were compared by the Paired test and independent sample t test.The correlations among various parameters were assessed by the Pearson linear correlation analysis.The differences of CCT and AST among different axial length groups were evaluated by one-way ANOVA.Results No significant differences were found in the CCT,CCV,axial length and AST between bilateral eyes (t =0.584,P =0.561 ; t =1.161,P =0.248 ; t =0.140,P =0.889 ; t =0.342,P =0.773).Temporal AST at 2 mm posterior to the sclera spur was (0.589 ±0.051)mm in the right eyes.An insignificant decline in CCT was found in the male group compared with the female group (right eyes:t =0.469,P =0.641 ; left eyes:t =0.465,P =0.643).However,compared with the female group,the increase of axial length,reduction of the mean CCV value and enhancement of the mean AST were observed(right eyes:all P＜0.01 ;left eyes:all P＜0.01).CCV showed a negative correlation with ocular axial length (r =-0.50,P＜0.01),but no significant correlation was found among age,CCT,ocular axial length and AST(P＞0.05).No remarkable differences were found in CCT and AST among the various axial length groups (CCT:F =0.998,P=0.372;AST:F=1.919,P=0.383).Conclusions In senile cataract patients,correlation is not found between AST and CCT;the increase of axial length is not associated with the thinning of the eyeball wall to a certain extent.Differences exist in some ocular parameters between different sexes.

Background A close relation between sclera thickness and glaucoma has been determined.Clinical features vary in different types of glaucoma patients,which hints that the scleral thickness might be distinct among these patients.Objective The aim of this study was to investigate the differences of central corneal thickness(CCT),anterior scleral thickness(AST) and axial length in glaucomatous patients.Methods This study was approved by the Ethic Commission of First Hospital of Peking University.Written informed consent was obtained from each individual prior to the examination.A retrospective descriptive study was designed.One hundred and sixty consecutive patients were recruited from March,2009 to November,2010 in First Hospital of Peking University,including 35 eyes with primary angle closure glaucoma(PACG) (35 cases),34 eyes with primary open-angle glaucoma POAG) (34 cases),37 eyes with normal tension glaucoma(NTG) (37 cases)and 17 eyes of ocular hypertension OHT) (17 cases).Thirty-seven eyes of 37 subjects with incipient cataract served as the control group.CCT and ocular axial length were measured with ultrasonic pachymeter and A-scan unit,respectively,and AST at the temporal quadrant 2 mm posterior to the scleral spur was measured by ultrasonic biomicroscopy (UBM).The measuring parameters among different groups were compared by analysis of covariance,and the correlations of AST,CCT with ocular axial length were analyzed using Pearson linear correlation and linear regression.The differences and correlation of CCT,AST and AL among five groups were analyzed.Results The CCT values in PACG group,POAG group,NTG group,OHT group and control group were (535.54 ± 5.20),(550.47 ± 5.28),(521.61 ± 5.07),(575.75 ± 7.76) and (535.06± 5.06) μm,respectively,showing a significant difference among them (F =9.560,P =0.000),and the CCT value of OHT group was increased in comparison with POAG group,PACG group,NTG group and control group(all P =0.000).The CCT of the POAG group was thicker than that in the PACG group,NTG group and control group(P=0.046,0.000,0.040).No significant difference was found in CCT among NTG group,PACG group and control group(P=0.950,0.060).The AST values of PACG group,POAG group,NTG group,OHT group and control group were(0.593±0.050),(0.600±0.050),(0.592-±0.060),(0.610-±0.060) and(0.604±0.060) mm,respectively,showing a insignificant difference among them (F =0.700,P =0.590).The axial length in the patients with PACG was shorter than that of the POAG group,NTG group,OHT group and control group (all P =0.000).The Pearson correlation analysis showed significantly positive correlation between CCT and AST in POAG group and NTG group(r=0.445,P=0.008;r=0.400,P=0.014).Conclusions This study confirms that there is dissimilarity in CCT but not in AST among different types of glaucomatous patients.The changes of CCT and AST are consistent in POAG and NTG patients.

Background Researches showed that microperimetry can exhibit more tiny visual function damage than conventional perimetry in glaucomatous eyes.However,the study on fixation stability of glaucoma is still rare until now.Objective This study was to compare the correlation between microperimetry Maia (Macular Integrity Assessment) and Humphrey perimetry,and to investigate the changes of the fixation stability in glaucoma patients with hemifield defect.Methods This study proposal was approved by Medical Ethic Committee of Peking University First Hospital.A cross-sectional study was performed under the informed consent of each subject.Thirtyfive eyes of 35 glaucoma patients with hemifield defect by 24-2 Humphrey perimetry were included in Peking University First Hospital from December 2013 to March 2014,and 30 eyes of 30 normal volunteers served as controls.Both Humphery (10-2) and Maia (expert 10-2) were performed on the subjects respectively and the correlation of the results between Humphery (10-2) and Maia (expert 10-2) were analyzed.Then the patients with normal hemifield on Humphrey were assigned to Maia normal group and Maia abnormal group.Fixation stability differences were compared between glaucoma group and normal control group,and between Maia normal group and Maia abnormal group.Results The moderately positive correlation was found in the mean sensitivity between Maia microperimetry and Humphrey perimetry (r=0.403,P =0.001),and the average threshold of Maia microperimetry was moderately positive correlated with the mean defect (MD) of Humphrey perimetry in glaucoma patients (r=0.438,P =0.008).The fixation stability parameter P1 was (67±17)％ and (87±10)％,and that of P2 was (70±16)％ and (88±9)％;the 63％ bicurve elipse area (BCEA) was (5.08±1.55) °2and (2.21±0.60) °2,and the 95％ BCEA was (14.74± 6.04) °2 and (2.86 ± 1.17)°2 in the glaucoma group and normal control group,respectively,showing significant decreases of P1 and P2 and increases of 63％ BCEA and 95％ BCEA in the glaucoma group compared with the normalcontrol group (t=-5.604,-4.831,9.885,11.086,all at P=0.000).In Maia normal group and Maia abnormal group,the P1 was (79±8)％ and (63±17)％,the P2 was (81±10)％ and (67±16)％,the 63％ BCEA was (3.19±0.65)°2 and (5.70±1.22)°2 and the 95％ BCEA was (9.10±2.60)°2 and (19.35±5.01)°2,respectively.Compared with the Maia normal group,the P1 and P2 were significantly lower,and 63％ BCEA and 95 ％ BCEA were higher in the Maia abnormal group (t=-2.468,P=0.019;t=-2.371,P=0.024;t =5.514,P=0.000;t=5.575,P=0.000).Conclusions Maia microperimetry and Humphrey perimetry yield a good correlation for glaucomatous macular function examination.In addition,Maia microperimetry showed that fixation stability decreased in glaucoma patients with hemifield defect.

Objective To study the association of TCF7L2 gene rs11196218,rs290487 polymorphisms with metabolic syndrome in type 2 diabetes mellitus population.Methods According to the diagnostic criteria of international diabetes federation (IDF),680 cases of type 2 diabetes patients were divided into metabolic syndrome (MS) group and non metabolic syndrome (control) group.DNA was extracted from peripheral mononuclear cells,and then PCR was performed to specifically amplify TCF7L2 gene fragments.Gene polymorphisms were determined by connected enzyme detection reaction.After population representative was checked by Hardy-Weinberg equilibrium,statistical analysis was completed by software SPSS 13.0.Results The population was accorded with Hardy-Weinberg equilibrium and possessed the population representative.Frequency distributions of genotypes (GG,AG and AA) in TCF7L2 gene rs11196218 in MS and control groups were 55.6％(233/419),35.8％(150/419),8.6％ (36/419) and 54.8％ (126/230),39.1％ (90/230),6.1％ (14/230),respectively.Frequency distributions of alleles(G and A) in TCF7L2 gene rs11196218 in MS and control groups were 73.5％(616/838),26.5％(222/838)and 74.3％(342/460),25.7％(118/460),respectively.Frequency distributions of genotypes (GG,AG and AA) in TCF7L2 gene rs290487 in MS and control groups were 14.8％(62/418),42.3％(177/418),42.9％(179/418) and 15.0％(34/226),48.2％(109/226),36.8％(83/226),respectively.Frequency distributions of alleles(G and A) in TCF7L2 gene rs11196218 in MS and control groups were 36.0％ (301/836),64.0％ (535/836) and 39.1％ (177/452),60.9％ (275/452),respectively.Frequency distribution of allele and genotype in TCF7L2 genes rsl 1196218 and rs290487 between the two groups were not associated with metabolic syndrome in type 2 diabetes population (P ＞ 0.05).Conclusions TCF7L2 gene rs11196218,rs290487 polymorphisms has not association with metabolic syndrome of type 2 diabetes.

Polyethylene glycol-polybenzyl-L-glutamate copolymer (PEG-PBLG) was synthesized and paclitaxel-loaded core-shell type nano-micelles with amphiphilic copolymer PEG-PBLG was prepared by the dialysis method. The drug loading content and entrapment efficiency were determined by HPLC. The average size and its distribution were determined by dynamic light scattering method. The paclitaxel release rate in vitro from micelles was measured by HPLC. The cell cytotoxicity in vitro was observed with MTT assay. The anti-tumor activity of paclitaxel-loaded micelles were evaluated in tumor-inhibiting test of nude mice using human liver cancer HepG-2. The results indicated that paclitaxel could be entrapped in PEG-PBLG copolymer micelles and its size was in the range of 80-265 nm which increased with an increase in molecular weight of PBLG in copolymer; in vitro the paclitaxel could be released sustainably from the micelles. In high concentration of paclitaxel (>20 microg x mL(-1)) the paclitaxel-loaded PEG-PBLG micelles displayed much less cell cytotoxicity than paclitaxel injections with Cremophor EL (P<0.05); the tumor inhibiting activity of paclitaxel-loaded PEG-PBLG micelles was similar to that of paclitaxel injections with Cremophor EL in the same paclitaxel concentration. It was concluded that the paclitaxel-loaded PEG-PBLG micelles had more uniform size and size distribution, excellent drug sustainable-release behavior, less cytotoxicity, good anti-tumor activity similar to paclitaxel injections with Cremophor EL. So paclitaxel-loaded PEG-PBLG micelles would be a novel paclitaxel preparation in clinic for the treatment of tumor.
Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Delayed-Action Preparations ; Drug Delivery Systems ; Humans ; Liver Neoplasms, Experimental ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Micelles ; Nanoparticles ; Neoplasm Transplantation ; Paclitaxel ; pharmacology ; Particle Size ; Polyethylene Glycols ; chemistry ; Polyglutamic Acid ; analogs & derivatives ; chemistry ; Polymers ; Random Allocation

Objective To establish a model for preparation of tissue-engineered skin grafts with hTERT cells carrying human papillomavirus type 6 (HPV 6) genome in vitro, so as to lay a foundation for studying HPV life cycle. Methods The full-length linear HPV6 genome and plasmid pEGFP-▲EGFP were electrophoretically cotransferred into hTERT cells. After selection using G418 resistance, Southern blotting was performed to determine the viral load of HPV6 in transfected cells. 3T3 J2 trophoblastic cells, type I rat-tail collagen and hTERT cells containing the full-length HPV6 genes (HPV6.hTERT cells)were mixed and cocultured on metal meshes to form skin graft-like structures. Hematoxylin and eosin (HE)staining was performed to observe the structure of formed skin grafts, an immunohistochemical assay to measure the expression of HPV6 L1 protein, and electron microscopy to observe virus particles in the skin grafts. Results The linear HPV6 gene was successfully transferred into hTERT cells, and Southern blotting showed the presence of HPV6 DNA in the transferred hTERT cells. The HPV6.hTERT cells, which were cocultured with 3T3 J2 trophoblastic cells and type I rat-tail collagen, proliferated and differentiated over time, and gradually formed skin grafts giving the appearance of verrucous hyperplasia. HE staining showed that the cocultured HPV6.hTERT cells could form typical stratified structure of skin after 7 days of cultivation, and histopathologic features of HPV infection, including obvious papillomatous hyperplasia, presence of vesicular cells, hyperkeratosis and parakeratosis, could be observed after 21 days. The immunohistochemical assay showed the expression of HPV6 L1 protein in the upper portion of skin grafts, and electron microscopy revealed the presence of HPV6 virus particles in skin grafts. Conclusions The established model for preparation of tissue-engineered skin grafts using HPV 6 genome-carrying cells provides a basis for biological studies of HPV, but its application is limited to some degree.

OBJECTIVETo investigate the heterozygous genotype and molecular characteristics of Organophosphorus resistance associated with heterozygous Estbeta2 of esterase B2 gene from natural population of Culex pipiens complex.

METHODSGenomic DNA was extracted from natural populations of Culex pipiens complex in Hangzhou. The PCR-restriction fragment length polymorphism (PCR-RFLP) assay was applied to type the resistance associated esterase gene. Estbeta2 of esterase B2 gene was identified by PCR-RFLP, and the genotyping for heterozygous Estbeta2 was carried out after restriction enzyme digesting by Bfm I endonuclease.

RESULTSThe DNA was isolated from 207 Culex pipiens respectively, while 156 PCR samples showed positive and the positive rate was 75.36% (156/207). The PCR-RFLP assay of esterase B2 gene revealed that the Estbeta2 was accounted about 28.20% (44/156) in 156 positive samples. There were two genotypes identified, namely homozygous Estbeta2 (90.90%, 30/33) and heterozygous Estbeta2 (9%, 3/33), heterozygous Estbeta2 was in existence of a hybrid form as which combined with Estbeta2 and a subtype (Estbeta2/Estbeta2(1)).

CONCLUSIONHeterozygous Estbeta2 of Organophosphorus resistance associated with esterase genotype was determined in natural population of Culex pipiens, and a genotyping method was established.

Animals ; Culex ; enzymology ; genetics ; Genes, Insect ; Genotype ; Heterozygote ; Insecticide Resistance ; genetics ; Insecticides ; pharmacology ; Organophosphorus Compounds ; pharmacology ; Phenotype ; Serine Endopeptidases ; genetics