ObjectiveTo investigate the proliferation and apoptosis effects of adenovirus-mediated interleukin-24 (Ad-IL-24) gene on Karpas299 cells in vitro. MethodsThe Karpas299 cells were divided into blank control group, Ad-IL-24 group, and the adenovirus which carrying green fluorescent protein gene group (Ad-GFP group). Karpas299 cells of Ad-IL-24 group were infected by adding 200.0 μL Ad-IL-24, Karpas299 cells of Ad-GFP group were infected by adding 200.0 μL Ad-GFP, but Karpas299 cells of blank control group were treated by adding 200.0 μL PBS. Cells' proliferation inhibition rates of 3 groups were detected by cell counting kit (CCK-8) method at 12, 24, and 48 hours after treatment, respectively, and the cells' apoptosis rates of 3 groups were detected by flow cytometry at 48 hours after treatment. ResultsAd-IL-24 can suppress the growth of Karpas299 cells, and the inhibition rate increased over time. Compared with Ad-GFP group at the same time, the cell' proliferation inhibition rate of Ad-IL-24 group was higher at 12, 24, and 48 hours after treatment (P<0.05). In addition, the cells' apoptosis rate of Ad-IL-24 group was higher than those of Ad-GFP group and blank control group at 48 hours after treatment (P<0.05). ConclusionAd-IL-24 can suppress the growth of Karpas299 cells and induce the apoptosis of it.
Objective To evaluate the host immune reaction against adenovirus mediated human bone morphogenetic protein 2 (Adv-hBMP-2) gene therapy in repairof tibial defects. Methods Twelve goats were made 2.1 cm segmental defects in he tibial diaphysis and divided into 2 groups. AdvhBMP2 transfected marrow mesenchymal stem cells(MSCs) and untransfected MSCs were implanted into the defect sites of transfected group(n=7) and untransfected group (n=5), respectively. The defect repair was observed by X-ray films after 4, 8, 16 and 24 weeks of transplantation and cellular and humoral immune reactions to adenovirus were assayed before implantation and after implantation. Results More bony callus was found in the bone defects of transfected group. The healing rates were 6/7 in transfected group and 2/5 in untransfected group, respectively at 24 weeks after implantation. The mixed culture of lymphocytes and MSCs showed that the lymphocytes stimulation indexes (SI) increased 14 days after implantation, and there was significant difference between the transfected group (4.213±1.278) and the untransfected group(－0.310±0.147，Plt;0.05); SI decreased after 28 days, but there was no significant difference between the transfected group (2.544±0.957) and the untransfected group (3.104±0.644，Pgt;0.05). After 14, 28, 49, and 120 days of treatment, the titer values of neutralizing antibody against Adv-hBMP-2 (log0.1) were 2.359±0226, 2.297±0.200, 2.214±0.215 and 2.297±0.210 in transfected group, and -0.175±0.335, -0.419±0.171, 0±0.171 and 0.874±0.524 in untransfected group, being significant differences betweentwo groups(Plt;0.05). Conclusion Adenovirus mediated BMP-2gene therapy can cause cellular and humoral immune reactions against adenovirus, which can eliminate the influence of adenoviral genes and proteins within a certain period.
ObjectiveTo analyze the effectiveness and problems existing in implementing acquired immune deficiency syndrome (AIDS) control and prevention in community health service centers, and to provide scientific evidence for promoting AIDS control and prevention. MethodsRelated information on AIDS control and prevention in community health service centers in a community in the whole year of 2013 was acquired. The effectiveness and problems existing in case management, laboratory testing and highly active antiretroviral therapy (HAART) based on the current community health service system were analyzed. ResultsBy the end of 2013, the rate of AIDS case management was from zero to 100%, the rate of CD4 T detection was from 76.60% (360/470) to 88.35% (508/575), and the rate of receiving highly active antiretroviral therapy among AIDS patients was from 81.40% (175/215) to 84.41% (287/340). But in the actual work on AIDS in community health service centers, there were still some common problems needing to be solved. ConclusionThe community level management mode of AIDS can promote the accuracy of AIDS patients' information, improve the rate of case management, the rate of CD4 T detection and HAART. It is suggested that the community level management mode of AIDS should be promoted in the cities where AIDS patients are concentrated.
ObjectiveTo describe the imaging and clinical features of vaccinia virus induced pneumonia by long-term follow-up.MethodsThe clinical data, imaging features and long-term follow-up of 5 patients with vaccinia virus pneumonia admitted to Wuxi People's Hospital Affiliated to Nanjing Medical University were analyzed.ResultsAll the 5 patients were male, aged between 21 and 54 years. The latent period of the disease was 2 to 5 days. All the patients had fever and pneumonia, while 3 of them had herpes. Two patients with severe pneumonia showed extensive patchy and nodular shadows in both lungs. Chest CT findings of the other three patients showed scattered small nodules in both lungs. All patients were followed up by telephone every half a year for 3 years. The prognosis of all patients was good. The patients reported in the English literature were clinically clustered, with fever, vomiting and rash as the main symptoms.ConclusionsVaccinia virus may cause different clinical symptoms through different transmission routes, and its infectivity is strong. Biological protection should be strengthened in laboratory and working environment.
Previous studies have shown that rehabilitation can relieve clinical symptoms, prevent complications, and improve physical function and quality of life in patients with pneumonia. This consensus summarizes the rehabilitation assessment, self-protection, rehabilitation goals, rehabilitation programs, indications for termination of treatment, nutrition support and psychological rehabilitation for patients with mild, moderate, severe and critical types of coronavirus disease 2019 based on the relevant literature and previous practice of coronavirus disease 2019. We hope this consensus can provide a reference for rehabilitation of coronavirus disease 2019.
【Abstract】ObjectiveTo investigate the relationship of magnetic resonance diffusion-weighted imaging (DWI) to histology in the patients of chronic viral hepatitis. MethodsThirty-five patients of chronic viral hepatitis who received liver biopsy and 10 healthy volunteers were included in this study. All of them underwent DWI on a 3.0T MRI device. Apparent diffusion coefficient (ADC) of the liver were measured respectively when b value were set as 100, 400, 600 and 800 s/mm2. Biopsy specimens were scored for fibrosis and necroinflammation according to the Knodell histology activity index (HAI). ResultsWhen b value was set as 800 s/mm2, statistical difference was showed between the fibrosis group and the nonfibrosis group, statistical difference was also shown among the different degrees of necroinflammation and fibrosis. ConclusionDWI is a valuable method for grading and staging of chronic viral hepatitis.
Objective To determine whether the transforminggrowth factor β1 (TGF-β1) is a key regulatory molecule required for an increase or a balance of extracellular matrix (ECM) and DNA synthesis in the goat passaged nucleus pulposus (NP) cells. Methods The NP cells isolated from the goat intervertebral discs were cultured in vitro for a serial of passages and transfected with the replicationincompetent adenoviral vectors carrying the human TGF-β1 (hTGF-β1) or lacZ genes. Then, they were cultured in monolayer or alginate bead 3dimensional (3-D) systems for 10 days.The changes in the production and the molecular components of ECM that occurredin the NP cells transfected with Ad/hTGF-β1 or the controls were evaluated by Westernblot and absorbance of glycosaminoglycan (GAG)-Alcian Blue complexes. Differences of DNA synthesis in the variant cells and culture systems were assessed by fluorometric analysis of the DNA content. ResultsA quantitation in the variant culture systems indicated that in monolayers the NP cells at Passage 3 transfected with Ad/hTGF-β1 had a much higher cell viability and more DNA synthesis(P＜0.05); however, in the alginate 3-D culture system, the NP cells transfected with Ad/hTGF-β1 did not have any significant difference from the controls(P＞0.05). The Western blotting analysis ofthe protein sample isolated from the variant cells for TGF-β1, type Ⅱ collagen, and Aggrecan expression indicated that in the monolayers and alginate 3-D culture systems the NP cells at Passage 3 transfected with Ad/hTGF-β1 revealed much higher protein levels than the controls(P＜0.05)； whereas the type Ⅰcollagen content was much lower than the controls (P＜0.05), but a significatly increased ratio of type Ⅱ/type Ⅰ collagen was found in both of the cell culture systems(P＜0.05). The GAG quantification also showed a positive result in both the cell culture systems and the NP cells at Passage 3 transfected with Ad/hTGF-β1 had a much higher GAG content than the controls(P＜0.05). Conclusion To a greaterextent, hTGF-β1 can play a key role in maintaining the phenotype of the NP cells and can still have an effect of the phenotypic modulation after a serial of the cell passages. The NP cells that are genetically manipulated to express hTGF-β1 have a promising effect on the restoration of the intervertebral disc defects. The NP cells transfected with Ad/hTGF-β1 cultured in the 3-D alginate bead systems can show a nearly native phenotype.
Since December 2019, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection has gradually spread all over the world. With the implementation of class B infectious disease management policy for coronavirus disease 2019 (COVID-19), China has experienced a pandemic. For patients receiving a time-sensitive or emergency surgery, SARS-CoV-2 infection may increase the risk of postoperative pulmonary complications. An appropriate perioperative mechanical ventilation strategy, such as lung protective ventilation strategy, is particularly important for preventing postoperative pulmonary complications in patients undergoing general anesthesia. In addition, how to protect medical personnel from being infected is also the focus we need to pay attention to. This article will discuss the perioperative mechanical ventilation strategy for COVID-19 patients and the protection of medical personnel, in order to provide reference for the development of guidelines.
Objective To research the transfer of adenovirus human bone morphogenetic protein 4 (Ad-hBMP-4) to human degenerative lumbar intervertebral disc cells in vitro and analyze its effect on the proteoglycan, collagen type II, and Sox9 of intervertebral disc cells. Methods Identified Ad-hBMP-4 was amplified and detected. Degenerative lumbar intervertebral disc cells were aspirated from the degenerative lumbar intervertebral disc of patients with Modic III level disc protrusion (aged, 27-50 years). The expressing position of collagen type II was identified in the intervertebral disc cells through the laser confocal microscope. The intervertebral disc cells at passage 1 were transfected with Ad-hBMP-4 as experimental group. After 3 and 6 days of transfection, RT-PCR was used to detect the mRNA expressions of proteoglycan, collagen type II, and Sox9, and Western blot to detect the expressions of proteoglycan and collagen type II proteins. Non-transfected cells at passage 1 served as control group. Results The virus titer of Ad-hBMP-4 was 5 × 106 PFU/mL. No morphological changes in the cells after transfection by Ad-hBMP-4. Collagen type II mainly expressed in the cell cytoplasm. The mRNA expressions of the proteoglycan, collagen type II, and Sox9 in experimental group at 3 and 6 days after transfection were significantly higher than those in control group by RT-PCR (P lt; 0.05), and the expressions of proteoglycan and collagen type II proteins were significantly higher than those in contorl group by Western blot (P lt; 0.05). There were significant differences between 3 days and 6 days in experimental group (P lt; 0.05). Conclusion Ad-hBMP-4 could transfect human degenerative lumbar intervertebral cells with high efficiency and promote collagen type II, proteoglycan, and Sox9 expressions. hBMP-4 may play an important role in the repair process during early disc degeneration.