【Abstract】ObjectiveTo construct a recombinant adeno-associated virus(rAAV2) vectors carrying the combined transcriptional regulatory sequences of α-fetoprotein enhancer and albumin promoter for the purpose of targeted gene therapy for hepatocellular carcinoma (HCC). MethodsThe fragment of combined transcriptional regulatory sequences of α-fetoprotein enhancer and albumin promoter was amplified through polymerase chain reaction (PCR) and cloned into the promoter site of pAAV-IRES-hrGFP instead of the CMV promotor in AAV Helper-Free System to construct the rAAV2 expression plasmid pAAV-IRES-hrGFP-EP. Then the packaging cell lines (HEK 293 cell) was co-transfected with the pAAV-IRES-hrGFP-EP together with the control plasmid pAAV-RC and pHelper in AAV Helper-Free System by means of lipofectamine.The recombinant adenoassociated virus vector(rAAV2-EP) carrying the combined transcriptional regulatory sequences of α-fetoprotein enhancer and albumin promoter was packaged and amplified in the HEK 293 cell. Then the viral titer was checked by GFP. ResultsThe recombinant adeno-associated virus vector(rAAV2-EP) carrying the combined transcriptional regulatory sequences of α-fetoprotein enhancer and albumin promoter was constructed successfully, the b green fluorescence was observed in HEK 293 cells under fluorescence microscope. The viral titer was 1.2×105. ConclusionConstruction of the recombinant adeno-associated virus vector rAAV2-EP driven by the combined transcriptional regulatory sequences of α-fetoprotein enhancer and albumin promoter would provide a sound basis and improved vector for targeted gene therapy for HCC.
Objective To evaluate the suitability of the biodegradable microsphere encapsulation of adenovirus as a targeting vector for gene therapy of hepatocellular carcinoma. Methods Encapsulate the recombinant adenovirus in PLG 〔poly (lactic/glycolic)〕 copolymer by the solution evaporation method, the release test and the bioactivity of viruses incorporated in vitro were studied. Results More than 19.3% of adenovirus was encapsulated in PLG microspheres. The release test shows that the adenovirus was released for more than 200 h, 50% were shed within the first 100 h, and their activity was retained. Conclusion Recombinant adenovirus can be formulated in a polymer preparation of PLG with retention of bioactivity. It may be a valuable vector for the gene therapy of liver cancer.
In this study, the role of newcastle disease virus (NDV) combined thermic solidified tumor vaccine in inhibiting growth of tumor and immune control was investigated, and rate of inhibiting tumor and cellular immunity were measured. The results showed that rate of inhibiting tumor in experimental group Ⅰ and Ⅱ were 24.8% and 41.1% respectively; average weight of tumor was significantly lower in both experimental groups than in control group, and activity of natural killing (NK) cells in experimental groups was higher than that in control group (Ｐ＜0.01). This suggests that NDV combined thermic solidified tumor vaccine can inhibit growth of tumor and improve activity of NK cells, and their effects are better than that of NDV.
Epstein-Barr (EB) virus infection is associated with various tumors of lymphoid and epithelial origin. EB virus exists in most humans as a latent infection. EB virus latent infection-related genes play a key role in the EB virus latent infection, and also play an important role in promoting the occurrence and development of related tumors. This article will briefly introduce the characteristics of EB virus latent infection, the protein coding genes and non-coding genes related to EB virus latent infection (including EB virus nuclear antigen genes, EB virus latent membrane protein genes, EB virus encoded small RNA genes and EB virus microRNA genes), and the main functional mechanism of these EB virus latent infection-related genes in EB virus latent infection and subsequent tumorigenesis. The purpose is to providea theoretical basis for a comprehensive understanding of the EB virus latent infection and the mechanism of tumors caused by EB virus.
目的:分析艾滋病患者抗病毒治疗后的临床疗效,比较不同基线CD4+T淋巴细胞计数增长情况。方法:纳入51例符合治疗标准的初治患者,采用国家标准抗病毒治疗一线方案和卫生部统一提供的免费药物,通过对服药后半月、1月、3月、6月、12月的时段进行临床评估和实验室检查,并比较不同基线CD4+T淋巴细胞计数水平治疗后的增长情况。结果:治疗12月后,各方案组疗效无差异,不同基线CD4+T淋巴细胞计数的增长有显著差异。毒副反应为肝损伤、过敏性皮疹,消化道反应为主。结论:HAART可显著的抑制体内HIV病毒的复制,重建机体的免疫功能,缓解患者病情,有利于存活期的延长。严重的毒副作用发生较少。
To construct the recombinant adeno-associated virus (rAAV) vector co-expressinghVEGF165 and hBMP-7 depending on internal ribosome entry site (IRES) sequence, to measure the virus titer and to ver ify the correct recombination. Methods The AAV helper-free system was used to generate the rAAV co-expressing hVEGF165 and hBMP-7 genes. The IRES sequence from the bicistronic eukaryotic expression plasmid pIRES was cut down and subcloned into the ITR/MCS containing vector pAAV-MCS to get pAAV-MCS A-IRES-MCS B, in which upstream MCS A and downstream MCS B was constructed. The hVEGF165 and hBMP-7 genes were ampl ified by PCR and inserted into MCS A and MCS B respectively. The recombinant expression plasmid pAAV-hVEGF165-IRES-hBMP-7 was co-transfected into AAV-293 cells with pHelper and pAAV-RC for packaging of recombinant AAV. The green fluorescent protein (GFP) labeled rAAVIRES GFP was simultaneously packaged by using the parallel plasmid pAAV-IRES-GFP. The efficiency of rAAV packagingwas monitored under fluorescent microscope and recombinant viral particles were harvested from infected AAV-293 cells.The virus titer was measured through infecting AAV-HT1080 cells, and the recombinant rAAV-hVEGF165-IRES-hBMP-7was verified by PCR of the exogenous interest genes of hVEGF165 and hBMP-7. Results The recombinant plasmid pAAVhVEGF165- IRES-hBMP-7 was verified by double digestion. Using the AAV helper-free system, GFP expression could be observed under fluorescent microscope 72 hours after triple plasmid co-transfection and the system provided a high packing ratio of 95%-100%. The rAAV has a high purity and high titer of 5.5 × 1011vp/mL, and AAV-HT1080 cell could be infected at a ratio of 90%. The recombinant virus was confirmed by PCR of exogenous hBMP-7 and hVEGF165 genes. Conclusion Re combinant rAAV-hVEGF165-IRES-hBMP-7 was successfully constructed with a high virus titer, which may offer the basement
At present, there are few studies on patients with epilepsy (PWE) and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), and the results of many studies are inconsistent. SARS-CoV-2 may cause new seizures through a variety of mechanisms, and the susceptibility and mortality of PWE to SARS-CoV-2 remains controversial. During the SARS-CoV-2 pandemic, anxiety, depression and other psychological problems were common among epileptic patients. Clinicians providing telemedicine, telephone chat and video call can effectively reduce the risk of psychological disorders in PWE patients. At the same time, there are many interactions between antiepileptic drugs and various antiviral drugs, which should be carefully considered when using. Considering that the research results are few and many studies contradict each other, PWE and SARS-CoV-2 deserve further exploration in future studies.
Objective To investigate the clinical characteristics and risk factors for perioperative lung surgery patients with SARS‐CoV‐2 Omicron variant infection. Methods The clinical data of patients who underwent lung surgery at the Department of Thoracic Surgery, Renmin Hospital of Wuhan University from December 1, 2022 to January 9, 2023 were retrospectively analyzed. The patients were divided into an infection group and a non-infection group according to whether they were infected with SARS-CoV-2. And the clinical data of two groups were collected and compared. Multiple linear regression analysis was used to explore the risk factors affecting the time of hospitalization. Results A total of 70 patients were enrolled in this study, including 36 (51.4%) males and 34 (48.6%) females at a median age of 61.0 (49.0, 66.8) years. There were 28 patients in the infection group and 42 patients in the non-infection group. The proportion of preoperative abnormal coagulation function and the risk of postoperative pulmonary infection in perioperative patients infected with SARS-CoV-2 were higher than those in the non-infection group (P<0.05). Subgroup analysis found that patients with preoperative SARS-CoV-2 infection were more likely to have pulmonary infection after surgery, but did not prolong the time of hospitalization or increase the risk of severe disease rate. The patients with postoperative SARS-CoV-2 infection had worse clinical prognosis, including longer time of hospitalization (P=0.004), higher ICU admission rate (P=0.000), higher lung infection rate (P=0.003) and respiratory failure rate (P=0.000). Multiple linear regression analysis showed that gender and extent of surgery were independent risk factors for prolonged hospitalization time. Conclusion Preoperative infection with SARS-CoV-2 Omicron variant will increase the risk of pulmonary infection, but it will not affect the clinical prognosis. However, postoperative infection with SARS-CoV-2 Omicron variant will still prolong the time of hospitalization, increase the ICU rate, and the risk of pulmonary complications.