Adult ; Calmodulin-Binding Proteins ; metabolism ; Desmin ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Leiomyoma ; metabolism ; pathology ; surgery ; Leiomyosarcoma ; pathology ; Spinal Canal ; Spinal Neoplasms ; metabolism ; pathology ; surgery ; Young Adult

AIM:To investigate the expression of microRNA-141 (miR-141) in human hepatocellular carcino-ma (HCC) cell line SMMC-7721 and normal hepatocyte line HL-7702, and to analyze the effect of abnormal expression of miR-141 on the malignant biological behaviors of human hepatocarcinoma cells.METHODS:The RNA from SMMC-7721 cells and HL-7702 cells was extracted.SYBR Green real-time PCR was performed to detect the expression of miR-141. Synthetic miR-141 mimic and its negative control were transfected into the SMMC-7721 cells, and miR-141 inhibitor and its negative control were transfected into the HL-7702 cells by the method of Lipofectamine.After transfection, MTS assay and BrdU-ELISA were employed to evaluate the effect of miR-141 on the cell proliferation.Flow cytometry was used to detect cell cycle and apoptosis.The changes of migration ability were investigated by Transwell invasion assay.RESULTS:The expression of miR-141 in the SMMC-7721 cells was significantly lower than that in the HL-7702 cells ( P<0.05 ) .Com-pared with blank group, Lipofectamine group and negative control group, the proliferation of the SMMC-7721 cells trans-fected with 25 nmol/L miR-141 mimic was significantly inhibited in a time-dependent manner (P<0.05).The percenta-ges of G1 phase cells and early apoptotic rate were significantly increased when miR-141 was up-regulated, but the migra-tion ability was inhibited (P<0.05).Compared with blank group, Lipofectamine group and negative control group, the proliferation of HL-7702 cells transfected with 50 nmol/L miR-141 inhibitor was significantly increased in a time-dependent manner (P<0.05).When miR-141 was down-regulated, the percentages of G1 phase cells and early apoptotic rate were significantly decreased, but the migration ability was enhanced (P<0.05).CONCLUSION:miR-141 is down-regulated in human hepatocarcinoma cell line.Up-regulation of miR-141 will not only inhibit cell proliferation and migration ability, but also affect the cell cycle and apoptosis of SMMC-7721 cells.miR-141 may function as a tumor suppressor gene during HCC development.

Objective To investigate the diagnosis and treatment value of laparoscopic surgery in tubal obstruction infertility patients with endometriosis.Methods Retrospective analysis was done in 381 cases underwent laparoscopic treatment for tubal obstruction from May 2005 to May 2010,,of which 119 cases were combined with endometriosis.The treatment results were analyzed,including the decomposition of the umbrella side tubal obstruction adhesions,lesions of endometriosis electrocoagulation,endometriosis cystectomy and pregnancy and so on.Results Hysterosalpingography(HSG)showed tubal occlusion in both side or one side in the distal end in 381 cases,and these patients were treated with laparoscopic surgery.There were 262 cases showed distal tubal occlusion not companied with endometriosis,of which 82 cases clinically pregnanced,accounting for 31.3％.Surgical treatment of endometriosis found in 119 cases,accounting for 31.2％ ; and 29 cases pregnaced,which accounted 24.4％.We found 42 cases of tubal adhesions grade Ⅰ-Ⅱ and endometriosis stage Ⅰ-Ⅱ,16 cases of clinical pregnancy,accounting for 38.1％ ;Twenty-six cases of tubal adhesions grade Ⅰ-Ⅱ and endometriosis stage Ⅰ-Ⅱ,7 cases of clinical pregnancy,accounting for 26.9％ ;Twenty-eitht cases of tubal adhesions grade Ⅲ-Ⅳ and endometriosis stage Ⅰ-Ⅱ,4 cases of clinical pregnancy,accounting for 14.3％ ;Twenty-three cases of tubal adhesions grade Ⅲ-Ⅳ and endometriosis stage Ⅲ-Ⅳ,2 cases of clinical pregnancy,accounting for 8.7％.The clinical pregnancy rate in the first group was significantly higher than the other groups,chnical pregnancy rate of the last group was the lowest.Conclusion Laparoscopic surgery is an effective treatment of tubal obstruction infertility combined with endometriosis.After laparoscopic surgery,the clinical pregnancy rate is significantly higher in Tubal obstruction without endometriosis than patients with endometriosis.After surgery,the clinical pregnancy rate in patients with the light stage tubal adhesions and endometriosis is better than those severely.

Benzene ; poisoning ; Female ; Humans ; Male ; Occupational Diseases

AIM:To investigate the changes of connexin 43 (Cx43) via establishing a model of sympathomi-metic atrial fibrillation ( AF) .METHODS:The mongrels ( n=15) were randomly divided into control group , rapid atrial pacing (RAP) group and isoprenaline (ISO) perfusion+RAP group (ISO+RAP group).All mongrels’ hearts were taken out rapidly by median sternotomy to establish the cardiac model with Langendorff perfusion in vitro.The atrial effective re-fractory period ( AERP) and AF inducability were tested .The expression and distribution of tyrosine hydroxylase ( TH) were analyzed by immunohistochemistry .Total protein level of Cx 43 and phosphorylation of Cx 43 were determined by West-ern blot.The distribution of Cx43 were also observed by immunofluorescence staining .The cell apoptosis was analyzed by TUNEL staining.The generation of reactive oxygen species ( ROS) in the mitochondria was measured by fluorescence spec-trophotometry .RESULTS:No significant change of AERP was found between control group and RAP group , while that in ISO+RAP group was significantly decreased (P<0.05) and induced AF.Compared with control group, the expression of TH, apoptotic index and the generation of ROS increased gradually (P<0.05), while the content of Cx43 decreased grad-ually both in the total protein and the phosphorylation levels in RAP group and ISO +RAP group (P<0.05).The fluores-cence intensity of Cx43 was also attenuated and Cx43 were lateralized apparently in RAP group , while Cx43 were character-ized as punctate distribution in ISO +RAP group.CONCLUSION:Sympathetic nerves may activate autophagosome at in-tercalated discs and trigger cell apoptosis , resulting in remodeling and downregulation of Cx 43 via oxidative stress , thus having effects on mediating and maintaining AF .

Objective Lamins are the major components of nuclear lamina underneath the inner nuclear membrane (INM).Lamins express in most cells and are involved in the whole process of growth, also play a major role in cell stability and embryonic development.Mutant in human LMNA gene may lead to a series of disorders, which are similar to progeria or other aging-associate syndrome.In this study, we report a new lmna knockdown animal model generated in our laboratory in order to provide a useful tool for studying laminopathies.Methods Two plasmids tagged to zebrafish lmna gene were designed based on morpholino oligonucleotides technology.Co-microinjected the plasmids into zebrafish embryos to knockdown lmna gene.Imagining and western blot detection were used to identify the mutants.Results Two different proteins, Lamin A/C, were expressed in the zebrafish embryos.Two plasmids lmna-MO and lmna-EGFP-pCS 2 + were generated and co-microinjected into embryos.The results of imagining and western blot showed that the expression of lmna gene was downregulated in the zebrafish embryos.Conclusions Lamin A/C are expressed in zebrafish.lmna gene can be knocked down by the injection of lmna-MO and lmna-EGFP-pCS 2 +.This new animal model may be a powerful tool for study on laminopathies.

Objective: To explore the relationship between NLRP3 inflammasome and atrial fibrillation (AF) by examining peripheral blood level of NLRP3 inlfammasome and other inlfammatory factors in relevant patients.
Method: A total of 60 AF patients were enrolled and divided into 2 groups: Paroxysmal AF (PAF) group and Non-paroxysmal atrial fibrillation (nPAF) group, n=30 in each group;in addition, there was a Control group including 26 healthy subjects from physical examination. NLRP3 expression in peripheral blood mononuclear cells (PBMCs) was measured by lfow cytometry;blood levels of IL-1β, IL-6, CRP and NT-proBNP were detected by ELISA. The correlations among different factors were studied by liner regression analysis and the differences were compared among groups.
Result:①Compared with Control group, PAF and nPAF groups had increased PBMCs level of NLRP3 and blood levels of IL-1β, IL-6, CRP, NT-proBNP, P<0.05, while NLRP3 level was similar between PAF group and nPAF group, P>0.05.②PAF and nPAF groups showed elevated blood level of NT-proBNP than Control group, P<0.05. ③PBMCs level of NLRP3 was positively related to left atrial diameter (r=0.579, P<0.05) and negatively related to left ventricular ejection fraction (r=-0.490, P<0.05) in both AF groups.
Conclusion: ① NLRP3 inflammasome was closely related to AF, which may provide a therapeutic target for AF treatment. ② AF was closely related to inflammatory response. ③ Downstream product of NLRP3 may cause the inlfammatory response which could induce the occurrence, development and maintenance of AF in relevant patients.

Objective Type A lamins are encoded by LMNA and a major component of the nuclear lamina, which have been suggested to play important roles in chromatin organization, transcription, DNA replication, and cell apoptosis. The aim of this study was to analyze the bioinformation of zebrafish lamins. Methods A phylogeny analysis was figured out with protein sequences of different species by Clustal X and MEGA 4?0 software. Then we compared the lamin protein sequences of different species with that of zebrafish by BLAST tool from NCBI. A figure of synteny analysis results was done with lamin sequence information of humans, murine and zebrafish cited from UCSC, Vega and Ensemble. Results The a?nalysis results showed that lmna, lmnb1, and lmnb2 genes of zebrafish are highly conservative and they may be homology of human LMNA, LMNB1 and LMNB2 genes. Conclusions Zebrafish lamins and human lamins have homologous sequence similarity, indicating that these two genes are orthologous genes.

Objective To choose zebrafish as the experimental animal model for studying the spatiotemporal expression rule of rap1 gen in zebrafish embryo early development process .Methods The Rap1 gene fragment was cloned from the zebrafish emby‐oscDNA ,then the Rap1 gene fragment and pCS2+ plasmid were performed the in vitro connetion and recombination was extracted , the combinant plasmid was correct after the double enzyme digestion ,colony PCR and sequencing identification .T3 RNA polymer‐ase in vitro transcription system was used to obtain the digoxin (DIG)‐labeled anti‐sense mRNA probe of Rap1 gene .The whole mount in situ hybridization method was adopted to detect the Rap1 expression in zebrafish embryo early development process . Results The positive hybridization signal of Rap1 gene was detected at the cell division junction region of 0 .75 hpf ,animal pole of 3 .70 hpf and 6 .00 hpf ,and notochord of 12 .00 -72 .00 hpf .Conclusion Rap1 gene might be involved in the early development process of notochord nervous system in zebrafish .

High temperature and humidity stress during seed growth and development of spring soybean can result in seed deterioration in South China. We isolated two genes (GmSBP and GmSBPL) encoding putative SBP proteins from soybean (Glycine max (L.) Merr.) to study their biological functions and response to abiotic stress,. The two SBP proteins are hydrophilic and incomplete membrane ones. Real-time quantitative (RT-PCR) analysis reveals that the expression of the two genes in the developing seeds of the seed deterioration resistant cultivar Xiangdou No. 3 and sensitive cultivar Ningzhen No. 1 was significantly affected by high temperature and humidity treatment. Meanwhile, the levels of sucrose and soluble sugar in the developing seeds of both cultivars were also affected under high temperature and humidity stress. During seed growth and development, the expression of the two genes as well as the levels of sucrose and soluble sugar reached the highest at 30 days after flower. GmSBP2 and GmSBPL were found to be differentially expressed in different soybean tissues. Sub-cellular localization indicated that two genes were located in cytoplasm and cell membrane. Our results indicate that GmSBP2 and GmSBPL might be involved in the response to abiotic stress, which will enrich our understanding of pre-harvest seed deterioration and resistance in soybean from one side.
China ; Gene Expression Regulation, Plant ; Genes, Plant ; Membrane Transport Proteins ; genetics ; Plant Lectins ; genetics ; Real-Time Polymerase Chain Reaction ; Seeds ; Soybean Proteins ; genetics ; Soybeans ; genetics ; Stress, Physiological