【Abstract】ObjectiveTo investigate the prophylactic effect of lamivudine monotherapy on the recurrence of hepatitis B after liver transplantation. MethodsThirtyone patients with hepatitis B related benign decompensated cirrhosis who underwent liver transplantation between February 1999 to June 2002 and survived more than 3 months were analyzed retrospectively. Lamivudine was administered to each patient after operation and some patients before operation for the prophylaxis of HBV recurrence. The HBV markers and HBV DNA in serum and bioptic liver tissues in all patients were evaluated before and after operation. ResultsTotal HBV recurrence rate was 19.4％（6/31） during average 38.2 months (3.2-70.2 months) follow up. HBV recurrence rate was 7.1％(2/28), 16.0％(4/25), 26.1％(6/23) and survival rate was 87.1％(27/31), 80.6％(25/31), 66.1％(20.5/31) after 1-, 3-and 5-year, respectively. One hundred milligram lamivudine administration peroral daily for 2 weeks prior to transplantation enable HBeAg 54.5％(6/11) and HBV DNA 50.0％(5/10) positive patients convert to negative respectively. ConclusionPreoperative administration of lamivudine monotherapy can effectively prevent allograft from HBV re-infection after liver transplantation. Lamivudine should be used to convert HBV DNA and HBeAg to negative.
目的:寻找对腺病毒灭活的有效方法,来预防大规模生产腺病毒时所致的生产细胞的污染。方法:采用紫外线照射和不同浓度过氧乙酸的灭活方法灭活腺病毒,并将处理后的病毒加入到正常人胚肾293细胞中培养,观察细胞是否出现病变作为灭活效果判断标准。结果:我们发现一定强度紫外线照射至少2 h以上才能完全灭活腺病毒,过氧乙酸达到1 %的浓度和至少作用30 min,才能将完全病毒灭活。结论:紫外线照射2 h以上能有效灭活腺病毒,我们可以用于操作台的腺病毒灭活;1%浓度的过氧乙酸作用30 min以上能有效将腺病毒灭活,我们可以运用于实验室大空间的腺病毒灭活。
Inpatients after COVID-19 infection, especially those admitted to intensive care unit (ICU), may encounter a series of coagulation dysfunction, which may lead to thrombosis, such as pulmonary embolism (PE), deep vein thrombosis (DVT) or arterial thrombosis (AT). Although there are many literatures on the incidence rate, prevention and treatment of venous thromboembolism (VTE) in hospitalized patients with COVID-19 infection, there are few data on the symptomatic and subclinical incidence rate of VTE after COVID-19 infection discharge. Therefore, there are no specific recommendations or guidelines for the prevention of VTE after discharge from hospital due to COVID-19 infection, and the current guidelines are controversial. In this study, we reviewed and summarized the existing literature on the incidence rate, prevention, diagnosis and treatment of venous thromboembolism in patients with COVID-19 infection, in order to provide guidance for VTE prevention in patients with COVID-19 infection after discharge.
摘要:目的: 探讨联合LCT和高危型HPV检测对CIN宫颈治疗后的随访意义。 方法 :对200例LCT异常,高危型HPV阳性,阴道镜活检证实为CIN1~3的患者行LEEP治疗或宫颈冷刀锥切,治疗后进行严格随访,包括LCT和高危型HPV检测,阳性病例行组织学检查。 结果 :(1)所有病例经治疗后均无病变残留,其治愈率为100%。(2)从治疗后3个月起,CIN1组高危型HPV转阴率为100%。在随访的第3个月和6个月,CIN2~3组高危型HPV转阴率分别为7317%和9085%,显著低于CIN1组,差异有统计学意义(〖WTBX〗P ＜005)。(3)从随访12个月起,一直有2例病例持续HPV阳性,均为CIN3患者,但LCT和阴道镜检查未发现细胞学异常,继续随访。 结论 :CIN治疗后高危型HPV的转阴时间及转阴率与CIN的级别有关;高危型HPV持续阳性,但LCT和阴道镜检查无异常者可继续严格随访;LCT联合高危型HPV检测是CIN治疗后临床追踪随访的有效手段。Abstract: Objective: To investigate the Significance of LCT joint highrisk HPV testing for followup after CIN treatment. Methods : 200 cases that highrisk HPV infection were tested by realtime PCR and CIN1~3 were confirmed with LCT and colposcopy biopsy were considered. The patients were treated with LEEP treatment or cold knife conization. After treatment, all cases were strictly followed up with LCT and HPV test, and the patients with positive results were examined by histology. Results : 1) After treatment, there was no residual disease in all cases, the cure rate was 100%. 2) From 3 months after treatment, highrisk HPV negative rate was 100% in CIN1 cases. While at 3rd and 6th month after treatment, highrisk HPV negative rate in CIN2~3 cases were 7317% and 9085%, which were significantly lower than those in CIN1 cases,the difference was statistically significant. 3) From the 12th monthafter treatment, there are still two cases of sustained highrisk HPV positive but normal with LCT and colposcopy biopsy. All cases are still strictly followedup. Conclusion : After treatment, the negative rate and time of highrisk HPV concerned with the grade of the CIN; the patients with persistent positive highrisk HPV, but without abnormalities detected by LCT and colposcopy biopsy could continue to strictly follow up; LCT joint highrisk HPV detection is an effective clinical means for followup after CIN treatment.
ObjectiveTo establish 16HBE cell lines stably expressing glutathione S-transferase mu 5 (GSTM5) gene, and explore the mechanism of GSTM5 nuclear translocation. MethodsRecombinant lentiviral expression vector containing GSTM5 gene was constructed and lentivirus was produced. After lentivirus infection of 16HBE cells, 16HBE-GSTM5 cell lines were obtained by screening with puromycin. Expression of GSTM5 in different cells was examined by RT-qPCR and Western blot. The nuclear translocation of GSTM5 was observed by confocal laser scanning microscope, after the 16HBE-GSTM5 cell lines were treated with tumor necrosis factor-α (TNF-α; 10 ng/ml) for 0.5 hour. ResultsLentiviral expression plasmids, PLVX-puro-3*flag-SBP-GSTM5-C and PLVX-puro-GSTM5-SBP-3*flag-N, were constructed and lentiviral particles were successfully packed. After infected with lentivirus and screened by puromycin, two cell lines, 16HBE-GSTM5-SBP-3*flag-N and 16HBE-3*flag-SBP-GSTM5-C, were obtained. GSTM5 expression in these two cell lines was significantly higher compared with the control group and parental cells. After treated with TNF-α for 0.5 hour, the nuclear translocation of GSTM5 in 16HBE-GSTM5-SBP-3*flag-N was much more obviously than that in 16HBE-3*flag-SBP-GSTM5-C. ConclusionThe N-terminal region of GSTM5 is critical for nuclear translocation induced by TNF-α, which is mediated by a novel and non-classical nuclear localization signal.
ObjectiveTo use failure mode and effect analysis (FMEA) to check and improve the risk of severe acute respiratory syndrome coronavirus 2 nucleic acid detection, and explore the application effect of FMEA in the emergency inspection items.MethodsFMEA was used to sort out the whole process of severe acute respiratory syndrome coronavirus 2 nucleic acid detection from January 30 to February 21, 2020. By establishing the theme, setting up a team, analyzing the failure mode and potential influencing factors. Then calculate the risk priority number (RPN), formulate preventive measures and implement continuous improvement according to the analysis results.ResultsA total of 2 138 cases were included. After improvement, the number of potential failure modes has been reduced by 2 (17 vs.19); the value of total RPN decreased (3 527.49 vs. 1 858.28). There was significant difference in average RPN before and after improvement [(185.66±74.34) vs. (97.80±37.97); t=6.128, P<0.001].ConclusionsIn the early stage of emergency inspection items, using FMEA can systematically check the risk factors in the process, develop improvement measures. It also can effectively reduce the risk of severe acute respiratory syndrome coronavirus 2 nucleic acid detection in hospital.
Objective To study the adenovirus-mediated human bone morphogenetic protein-2 gene （Ad-hBMP-2）transferred to the intervertebral disc cells of the New Zealand rabbit in vitro. Methods The cells of New Zealand white rabbitswere isolated from their lumbar discs. The cells were grown in the monolayer and treated with an adenovirus encoding the LacZ gene (Ad-LacZ) and Ad-hBMP-2 (50,100, 150 MOI,multiplicity of infection) in the Dulbecco’s Modified Eagle Medium and the Ham’s F-12 Medium in vitro. Three days after the Ad-hBMP-2 treatment,the expression of hBMP-2 in the cells that had been infected by different dosesof MOI was determined by immunofluorescence and the Western blot analysis, and the expression was determined in the cells with the Ad-LacZ treatment in a dose of 150 MOI. Six days after the Ad-hBMP-2 treatment, mRNA was extracted for the reverse transcription polymerase chain reaction (RT-PCR) and the difference was detected between the control group and the culture group that was treated withAd-hBMP-2 in doses of 50, 100 and 150 MOI so that the expressions of aggrecan and collagen ⅡmRNA could be observed. Results The expression of hBMP-2 in the cells was gradually increased after the transfection in an increasing dose, which was observed by immunofluorescence and the Western blot analysis. At 6 days the aggrecan and collagen type Ⅱ mRNA expressions were up-regulated by Ad-hBMP-2 after the transfection at an increasing viral concentration in the dosedependent manner. Conclusion The results show that Ad-hBMP-2 can transfect the rabbit intervertebral disc cells in vitro with a high efficiency rate and the expression of hBMP-2 after theinfection is dose-dependent in the manner. AdhBMP-2 after transfection can up-regulate the expression of aggrecan and collagen Ⅱ mRNA at an increasing viral concentration.
【Abstract】Objective To explore the effect against gastric cancer induced by Newcastle disease virus modified autologous tumor vaccine （NDV-ATV）pulsed dendritic cells(DCs). Methods The Newcastle disease virus infected the gastric cancer lines (MNK45) and was lost its activity. Peripheral blood mononuclear cell (PBMC) were cultured under condition of recombinant human granulocyte macrophage-colony stimulating factor (1 000 u/ml)+IL-4(1 000 u/ml) + TNF-α(100 ng/ml). The tumor antigen specific cytotoxic T lymphocytes (CTL) was generated from activated autologous T cell by the Newcastle disease virus infected the MNK45 pulsed DC. And Cyto Tox 96TM in vitro assayed the cytotoxicity of CTL to MNK45. Thawed gastric cancer cell antigen were used as control in these experiments. Results The killing rate of MNK45 by antigen specific CTL reached (90.15±9.82)%, which was nearly twice as high as that of control(60.57±5.74)%. The CTL had much higher cytotoxicity to different differentiated type of gastric cancer cells such as MGC803〔(52.23±6.45)% 〕 and SGC7901〔 (61.75±8.84)%〕, as compared with LOVO〔(9.11±3.42)%〕 and HepG2 〔 (8.30±3.12)%〕tumor cells(P＜0.05). Conclusion Efficient and specific of against gastric cancer immunoreaction can be induced in virtue of NDV-ATV pulsed DCs, NDV-ATV loaded DCs might provide a new kind of theraputic means for gastric cancer.
目的:探讨丙型肝炎病毒非结构蛋白NS4B在原发性肝癌发生中的作用,及其发生机制。方法:设置对照组、空白载体PCXN2组、转染NS4B组。使用脂质体介导法,转染丙型肝炎病毒非结构蛋白重组质粒NS4B进入Chang肝细胞内,并用G418筛选。绘制生长曲线,分别用流式细胞仪检测瞬时表达及稳定表达时肝细胞凋亡率,用均数±标准差表示,统计学分析采用Dunnett t检验及q检验。结果:瞬时表达各组(PCXN2,NS4B)凋亡率分别为:(918±060)%,(445±053)%。稳定表达各组(PCXN2,NS4B)凋亡率分别为:(1575±209)%,(366±034)%。与PCXN2组比较Plt;001。结论:NS4B抑制肝细胞凋亡率,可能导致肝细胞异常增殖,诱导肝癌发生。