Objective To explore the clinical effect of local prophylactic application of methotrexate(MTX) in laparoscopic linear salpingostomy. Methods The data of 102 women with confirmed unruptured tubal pregnancy and desire to conceive were collected.They were divided into two groups: the study group(local application of MTX in the operation,n=51) and the control group(without application of MTX,n=51).The demographic data,the incidence of persistent ectopic pregnancy(PEP)postoperation,and the hysterosalpingography(HSG) 3 months(postoperation) were analysed,and the side effects of the study group were observed. Results There was 1 case of PEP in the study group and 9 cases in the control group(2.0% vs 17.6%,P0.05).There were no obvious side effects such as nausea,vomit and dental ulcer in the study group. Conclusion Patients who were treated with local MTX injection during the laparoscopic linear salpingostomy present less incidence of PEP with no obvious side effects.

Objective:To investigate the effect of Rab7 on cytokine induced by TLR7 (Toll like receptor-7) R848 activated in Raw264.7,and discusses the influence of Rab7 on MAPK signal transduction.Methods: TLR7 downstream cytokines such as TNF-α,IL-6,IFN-α,IFN-β and IP-10 activated by R848 were detected through Q-PCR in Rab7 silenced mouse macrophages,and then analysis of phosphorylation of MAPK determined with Western blot showed the effect of Rab7 on signal transduction of MAPK.Results: Rab7 inhibit production of cytokine activated by TLR7,and also,Rab7 had an inhibitory effect on MAPK signal pathway.Conclusion: The experimental results further illustrate that the Rab7 is the TLR7 signal transduction pathway negative regulatory factor,and to participate in MAPK signaling pathway.

Objective:To establish cell lines stably expressing Rab5a and its the inactive mutant Rab5aN133I,analyze the effect of Rab5a on the expression of cytokines in LPS-stimulated RAW264.7 cells .Methods:RAW264.7 cells were transfected with Rab5a and its inactive mutant vector Rab5a N133I separately,and then screened by G418.Rab5a stable expressing cell lines were identified by Real time-PCR.The growth of the stable cell lines was analyzed by MTT assay.After the stable cell lines were stimulated by LPS for different time periods,the expression of iNOS,TNF-αand IL-6 was detected.Results:Rab5a and Rab5aN133I transfection resulted in elevated Rab5a mRNA expression compared with the control cells ( P<0.05 ).Rab5a overexpression enhanced the proliferation of RAW264.7 cells.However,the proliferation of Rab5aN133I cells was significantly slower than the control cells ( P<0.05).Overexpression of Rab5a promoted LPS-induced production of iNOS,TNF-αand IL-6 in RAW264.7 cells (P<0.01). Conversely,overexpression of Rab5aN133I abolished the stimulating effects of Rab5a.Conclusion: Rab5a promoted LPS-induced expression of iNOS,TNF-αand IL-6 in RAW264.7 macrophages in a GTP-binding ability-dependent manner.

Objective To explore the infection and drug sensitivity of ureaplasma urealyticum(Uu)and mycoplasma hominis (Mh)in vaginal secretion of gynecological outpatients.Methods The infection and drug sensitivity test of Uu and Mh in va-ginal secretion samples of 1 800 patients collected from January 2015 to April 2016 were detected with mycoplasma culture i-dentification and counting drug sensitivity kit produced by Zhengzhou Antu Luke Bioengineering Co.,Ltd.Results The positive rate of Uu(57.27%)was significantly higher than that of Mh(2.78%,χ2=33.69,P<0.001).The positive rate of mycoplasma was highest in the age group of 31~35(77.09%),but that of Uu was highest in the age group of 21~25 (65.83%)and that of Mh in 36~40 years old group(9.09%),in addition that of multiple infection by Uu and Mh was highest in less than 20 years old group(20.51%).There were statistical difference for Un,Mh and co-infection by Un and Mh between age groups(χ2=15.505~36.574,P<0.01).The top three drugs sensitive for mycoplasma were josamycin, minocycline and doxycycline and that last three ones were clindamycin,thiamphenicol and sparfloxacin.The drug sensitive rates for 12 antibiotics against Uu were higher than those against co-infection by Uu and Mh,but those of erythromycin,gat-ifloxacin,azithromycin,clarithromycin and Luo Hongmei against Mh were lower than those against co-infecion of Uu and Mh.Conclusion The detection of mycoplasma and drug sensitivity in vaginal secretions provides the experimental basis for clinical diagnosis and treatment.

OBJECTIVETo analyze the feasibility of reverse island flaps pedicled with cutaneous nerve nutrient vessels in repairing the defect on distal parts of extremities.

METHODSThirty patients with tissue defect in distal extremities, complicated by exposed vessels, nerve, tendons, and/or bones ,were repaired with island flaps pedicled with neurocutaneous nutrient vessel. Among them, five cases were grafted with flaps with external forearm cutaneous nerve vessels, eleven cases were grafted with flap containing sural neural nutrient vessels in its pedicle,and 14 cases with saphenous nerve nutrient vessels in the pedicles.

RESULTSThe flaps of 28 cases survived with perfect appearance and function. In one case partial necrosis occurred because of compression on the pedicle, but the patient healed after dressing, and another case with necrosis of the edge of the flap due to infection, but also healed after skin grafting.

CONCLUSIONThe reverse island flaps pedicled with cutaneous nerve nutrient vessels which are constant in anatomy, with reliable blood supply, can be recommended because its simple operative technique, non-injurious to main blood vessels and muscles, the repair of distal defects of the extremities.

Adolescent ; Adult ; Child ; Extremities ; injuries ; Humans ; Male ; Middle Aged ; Peripheral Nervous System ; Reconstructive Surgical Procedures ; Skin ; innervation ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; innervation ; Wound Healing ; Young Adult

Objective:To examine the distribution of syphilis antibody in pregnant women and newborns and to explore how to optimize the existing syphilis screening process by setting the diagnostic gray area.Methods:The results of syphilis testing obtained from 119 531 pregnant women and 21 275 newborns from 2015 to 2018 by automatic chemiluminescent immunoassay (CLIA) and the re-examination results determined by Treponema pallidum particle agglutination (TPPA) and the rapid plasma reagin test (RPR) were retrospective analyzed. Data analysis was performed by Chi-square, Fisher′s exact test and Chi-square test for trend. Results:The positive rates of Syphilis specific antibody (TPAb) in clinical specimens from pregnant women and newborns were 0.69% (825/119 531) and 1.24%(264/21 275). The total re-examination positive rates were 0.32% (380/119 531) and 0.90%(191/21 275), and the suspicious syphilis prevalence rates in these specimens were 0.13% (161/119 531) and 0.31%(67/21 275), respectively. The suspicious syphilis prevalence rates in specimens of pregnant women from 2015 to 2018 and newborns increased year by year (χ 2=9.860, P=0.002; χ 2=5.311, P=0.021). With the elevation of the optical density value of samples to cut-off ratio (S/CO) value, positive coincidence rate of TPPA and TPAb in pregnant women and newborns increased significantly (χ 2=614.833, P<0.001; P<0.001). When the S/CO value in newborns exceeded 7.00 or the S/CO value in pregnant women exceeded 15.00, the effectiveness of TPAb results is equivalent to TPPA. The prevalence of suspected syphilis in pregnant women and newborns also increased with the increase of S/CO value (χ 2=323.059, P<0.001; P<0.001). When the S/CO value in newborns bellowed 3.00 or the S/CO value in pregnant women bellowed 5.00, the prevalence rate of suspected syphilis was 0%, which could preliminarily exclude syphilis infection. Conclusions:The prevalence rates of suspected syphilis in pregnant women was increasing during the recent years. It is necessary to further strengthen syphilis screening and intervention treatment in early pregnancy to improve the rate of eugenics. Being a primary screening method for syphilis in pregnant women and newborns, CLIA has high false positive rate. According to the gray area established in this study, the syphilis screening process can be optimized to prevent missed detection, which may reduce the false positive rate and avoid clinical misdiagnosis.

BACKGROUNDTo survey a diarrhea outbreak in Guangan city and analyze the cause of the disease.

METHODSThe population enrolled in the surveillance came from four different settings and was randomly sampled. Stool specimens collected from diarrhea patients were tested ordinarily for enteric bacteria and further examined for viral pathogens with PAGE, ELISA and RT-PCR.

RESULTSIn total, 4,567 persons were surveyed, among them 942 had acute diarrhea (prevalence 20.63%). The incidence was higher in rural area (28.6%) than in urban area (19.6%) (chi-square =22.29, P less than 0.005) with a peak in May 10 through 25 four human caliciviruses were detected from stool specimens by ELISA and RT-PCR in specimens from 4 and 1 patients, respectively.

CONCLUSIONHuman calicivirus probably was the cause of this diarrhea outbreak in Guangan city.

Acute Disease ; epidemiology ; Adolescent ; Adult ; Age Distribution ; Caliciviridae Infections ; epidemiology ; virology ; Child ; China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Disease Outbreaks ; Feces ; virology ; Female ; Humans ; Male ; Middle Aged ; Norovirus ; isolation & purification ; Young Adult

OBJECTIVETo research the effects of glycogen synthase kinase (GSK3β) overexpression and GSK3β inhibitor SB-216763 on the proliferation of hepatic oval cells in rats and its regulatory mechanisms by Wnt signaling pathway.

METHODSThe hepatic oval cells WBF-344 were divided into the blank control group, GSK3β over-expression group, DMSO control group and GSK3β inhibitor groups, while the inhibitor groups set up three concentration gradients, that was 1, 5, 10 µmol/L. Using the GSK3β over-expression lentivirus, which had been identified correctly, and SB-216763 dealt with the cells WBF-344. The cells morphology of each group was observed under the phase contrast inverted microscope, and the expression of fluorescence in the lentivirus-transfected group was observed under the fluorescent microscope. The proliferation of each group cells was tested by CCK8 kits. The cells' apoptosis was detected by AnnexinV-FITC/PI kits. The expression of GSK3β, β-catenin and cyclin D1 were detected by Western blot.

RESULTSThe cells of GSK3β over-expression group were fewer and obvious aging. However, in each inhibitor added group, the cells' division and proliferation was vigorous, and the condition was good. Moreover, the cells' proliferation was getting stronger with the concentration of SB-216763 increasing. A large number of green fluorescence was expressed in the lentivirus-transfected cells. The cells' proliferation in GSK3β over-expression group restrained (t = 7.178, P < 0.01, as compared with control), while the cells' proliferation was vigorous in inhibitor groups (F = 45.030, P < 0.01, as compared with control). Flow Cytometry showed that the cells apoptosis was significant in GSK3β over-expression group. Western blot showed that the expression of GSK3β was increased, while the expression of β-catenin and cyclin D1 was decreased in the over-expression group. The expression of GSK3β had no significant difference among the control group and inhibitor groups. However, the expression of β-catenin and cyclin D1 was significantly increased with the concentration of SB-216763 increasing.

CONCLUSIONSThe overexpression of GSK3β can inhibit the Wnt signaling pathway, thus restrain the cells' proliferation and promotes apoptosis. SB-216763 can activate the Wnt pathway, thus promotes cells' proliferation.

Animals ; Cell Line ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Glycogen Synthase Kinases ; metabolism ; Hepatocytes ; drug effects ; Indoles ; pharmacology ; Male ; Maleimides ; pharmacology ; Rats ; Transfection ; Wnt Signaling Pathway ; beta Catenin ; metabolism

OBJECTIVETo investigate the effects and related mechanisms of hepatitis B virus X (HBx) protein on cell cycle and growth in hepatocellular carcinoma.

METHODSA human hepatocyte HepG2 cell line stably expressing a green fluorescent protein (GFP)-tagged HBx (HepG2/GFP-HBx cells) was used for the experiment, and HepG2 parental and HepG2/GFP cells was used as the controls. Effect of HBx on cell growth was evaluated by the MTT cell proliferation assay and on cell cycle progression by flow cytometry analysis of cells with or without treatment with 5-aza-2'-deoxycytidine (5-Aza-CdR; 5 pmol/L). Effect of HBx expression on promoter methylation status of the p16INK4A tumor-suppressor gene was detected by methylation-specific polymerase chain reaction and on p16 protein level was analyzed with western blotting.

RESULTSThe HepG2/GFP-HBx cells showed significantly higher cell proliferation at 72 hrs of culture (3.225+/-0.038 A490) than either control (HepG2: 2.012+/-0.022 A490, t = -46.86, P less than 0.001; HepG2/GFP: 2.038+/-0.029 A490, t = 42.51, P less than 0.001). The HepG2/GFP-HBx cells also showed significantly lower proportion of cells in the G0/G1 phase (16.45%+/-0.45%) than either control (HepG2: 44.81%+/-1.36%, t = -34.202, P less than 0.001; HepG2/GFP: 42.76%+/-1.58%, t = -28.88, P less than 0.001). However, 5-Aza-CdR treatment did lead to a significant amount of HepG2/GFP-HBx cells being arrested in the G0/G1 phase (33.25%+/-0.79%, t = 31.85, P less than 0.001). The p16INK4A promoter was methylated in the HepG2/GFP-HBx cells, and became demethylation after treatment with 5-Aza-CdR. However, no methylation of p16INK4A promoter was observed in both HepG2 and HepG2/GFP cells. The p16 protein level was significantly lower in the HepG2/GFP-HBx (vs. HepG2 and HepG2/GFP cells) and this level increased after treatment with 5-Aza-CdR.

CONCLUSIONHBx protein promotes hepatocellular carcinoma cell cycle progression and growth by shortening the G0/G1 phase, and the underlying mechanism may involve inducing p16INK4A promoter methylation and downregulating p16 protein expression.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Genes, p16 ; Hep G2 Cells ; Hepatitis B virus ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Promoter Regions, Genetic ; Trans-Activators ; pharmacology

This study was aimed to investigate the expression difference of serum cytokines in 20 patients receiving HLA-identical nonmyeloablative allogeneic hematopoietic stem cell transplantation (iNAHSCT) and HLA-haploidentical nonmyeloablative allogeneic hematopoietic stem cell transplantation (hiNAHSCT). IL-2, IL-4, IL-6, IL-10, TNF-α, γ-IFN and IL-17 were detected by flow cytometric bead array before and on week 1, 2, 4 after transplantation respectively. The results showed that the IL-2 level was found to be up-regulated at week 1 and 2 after transplantation in iNAHSCT group and in hiNAHSCT group respectively, but there was no difference between these two groups (P > 0.05). The γ-IFN levels was up-regulated at week 4 after transplantation in above-mentioned two groups, but no difference was found between these two groups. The IL-4 level increased at week 2 and 1 after transplantation in iNAHSCT and hiNAHCT groups respectively, but the IL-4 level in iNAHSCT group was higher than that in hiNAHSCT group. The IL-6 level rose at week 1 and 2 after transplant in above mentioned groups respectively, and reached to peak level at week 4 after transplantation, but IL-6 level in hiNAHSCT was higher than that in iNAHSCT group (P < 0.02). The IL-10 level was up-regulated at week 1 and 2 in iNAHSCT and hiNAHSCT groups respectively, but the IL-10 level in iNAHSC was higher than that in hiNAHSCT group. The TNF-α level was up-regulated at week 1 in hiNAHSCT group, but at week 2 in iNAHSCT group after transplantation. The TNF-α level in hiNHASCT group was higher than that in iNAHSCT group (P < 0.01). The IL-17 level was up-regulated at week 1 and week 4 after transplantation in hiNAHSCT and iNAHSCT groups respectively, the IL-17 level in hiNAHSCT group was high as compared with that in iNAHSCT group. It is concluded that the serum cytokine levels are obviously up-regulated in iNAHSCT and hiNHASCT groups, and reach to peak level at week 4 after transplantation. The IL-6, TNF-α and IL-17 level up-regulated significantly in hiNAHSCT group, but the IL-4 and IL-10 level up-regulated significantly in iNAHSCT.
Adolescent ; Adult ; Child ; Cytokines ; blood ; Female ; HLA Antigens ; immunology ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Interleukin-10 ; blood ; Interleukin-17 ; blood ; Interleukin-4 ; blood ; Interleukin-6 ; blood ; Male ; Middle Aged ; Transplantation, Homologous ; Tumor Necrosis Factor-alpha ; blood ; Young Adult