Objective:To construct a RNAi lentiviral vector targeting rat CD80 gene and detect its effect of gene silencing in NRK and IEC6 cells.Methods:The effective sequence of siRNA targeting rat CD80 gene was confirmed in our previous work.Oligo-DNA fragment containing short hairpin frame was synthesized and reannealed,and then cloned into pGCSIL-GFP lentiviral expression vector.PCR and sequencing analysis were made for verifying the positive clones.The virus packaging plasmids were transfected into 293T cells to harvest shRNA lentivirus.After infection in NRK and IEC6 cells,Real-time PCR was performed to determine the expressing level of CD80.Results:PCR and sequencing revealed that shRNA plasmids was correctly constructed.Virus with a titer of 4×10~8 TU/ml was successfully packaged.CD80 expression in NRK and IEC6 cells could be knockdown by virus infection as characterized by 66.9% and 63.5% decrease of CD80 mRNA in NRK and IEC6 cells respectively,compared with negative control lentivirus.Conclusion:The recombinant lentiviral shRNA expressing vector targeting rat CD80 gene has been successfully constructed and packaged.CD80 mRNA could be down-regulated availably in NRK and IEC6 cells.

OBJECTIVETo investigate the influence of interferon-alpha on the variation of cytotoxic T lymphocytes (Tc) and suppressor T lymphocytes (Ts) in the peripheral blood of 32 patients with chronic hepatitis B, and to analyse the relationship between the efficacy of interferon-alpha and the variation of Tc and Ts cells.

METHODSThe peripheral blood Tc and Ts cells were detected by the double-staining immunocytochemistry technique.

RESULTSAt the end of the treatment with interferon-alpha, there were 9 complete responders (CR), 12 partial responders (PR) and 11 non-responders (NR). Tc cells significantly increased and Ts cells markedly decreased in CR or PR group compared with the healthy control group. There was no significant difference in the level of Tc and Ts cells between CR and PR groups, and no significant difference in the level of Tc cells in NR, CR and PR groups, The Ts cells was significantly higher in NR group than in CR or PR group.

CONCLUSIONSThe treatment of interferon-alpha can result in the change of Tc and Ts cells in patients with chronic hepatitis B. The variation of Ts cells may play an important role in the efficacy of interferon-alpha against hepatitis B virus.

Adult ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Lymphocyte Count ; Male ; Middle Aged ; T-Lymphocytes, Cytotoxic ; cytology ; drug effects ; T-Lymphocytes, Regulatory ; cytology ; drug effects ; Time Factors ; Treatment Outcome

The expression of TNF-α in the liver at different periods post Schistosoma japonica infection and the effect on liver fibrosis after supplementary injection of these eytokines were investigated. The mice infected with schistosome cercariae were divided into 3 groups: normal control group, TNF-α-untreated infection group and TNF-α-treated infection group. ABC immunohistochemistry and pathologic image multimedia quantification system were applied to dynamically detect the activity of TNF-α. The results showed that the levels of TNF-α in the liver in TNF-α-untreated infection group were slowly decreased with prolongation of infection time (from 8th, 11th, 14th to 18th week), while in the TNF-α-treated infection group, those were increased significantly after intraperitoneal injection of TNF-α at 6th week after infection. At first to 8th week after the final injection of TNF-α, the intrahepatic TNF-α levels in the TNF-α-treated infection group were significantly higher than in the other two groups (P＜0.01), and the granulomatous inflammation and fibrosis in the liver were also milder in the normal control group. It was concluded that at the early stage of Schistosoma japonica infection mouse liver mainly released Th1 cytokine and TNF-α from Th1 activated macrophages. Six weeks after infection (post egg deposition), exogenous supplement with intraperitoneal injection of TNF-α could induce the enhanced expression of Th1 cytokines and alleviate the liver granulomatous inflammation and fibrosis.

Activity of IL-1 produced by peripheral blood monocytes stimulated with lipopolysaccharide in vitro was detected from 17 acute cases, 14 chronic cases and 19 advanced cases of schistosomiasis japonica. It was found that the level of IL-1 was significantly increased and positively related to the body tempereture in the group with acute schistosomiasis. The activity of IL-1 was statistically reduced in the chronic and advanced groups, especially in the latter. After inhibiting the synthesis of prostaglandin with indomethacin, the level of IL-1 was significantly increased in three groups of patients, but no apparent change in the normal contro group. The results indicate that IL-1 may play an important role in inducing the inflammatory reaction in patients with acute schistosomiasis japonica and in the immunoregulation in the chronic stage. The changes of IL-1 activity in patients with schistosomiasis japonica may be closely related to prostaglandin.

Objective:To observe the impact of core stability training in the sling exercise therapy pattern on the balance of stroke survivors.Methods:Sixty stroke survivors were randomly divided into a control group ( n=30) and an experimental group ( n=30). In addition to routine rehabilitation training, the control group received traditional core stability training, while the experimental group underwent core stability training applying the sling exercise therapy pattern. Before and after 4 weeks of training, the standing balance of both groups was evaluated using the Prokin balance trainer, with the length and area of motion recorded when performing the balance test with the eyes open and closed. Surface electromyography was used to record the average EMG (AEMG) values of the bilateral erector spinae and multifidus muscles during the balance testing. Results:After the 4 weeks of training the length and the area of progression of the center of the pressure were both significantly smaller for both groups than before the training with the eyes both open and closed. The average length and area in the experimental group were significantly less than among the controls. The AEMG values recorded during the balance tests were significantly higher than those before the intervention for both groups, with the multifidus muscle averages on the affected side significantly greater in the experimental group than among the controls when performing the balance test with the eyes closed.Conclusion:Core stability training in the sling exercise therapy pattern is superior to conventional core stability training because it can better improve the balance of stroke survivors.

Deepening the ideological and political construction of curriculum and carrying out the fundamental task of cultivating people with morality are the important requirements of education reform and talent cultivation in the new era. Microbiology Experiment is an important basic course and core practice course of Bioengineering, Pharmaceutical Engineering, Food Science and Engineering, et al. In order to give full play to the education function of Microbiology Experiment, this article deeply developed the ideological elements contained in the curriculum referring to the guidelines for the construction of ideological and political courses in institutions of higher education. And the article explored the ideological and political reform of Microbiology Experiment from three aspects: teaching content reform, teaching method innovation and improvement of teachers' ideological and political construction ability. Strive to integrate the value shaping, knowledge transference and ability training, cultivate high-quality professionals with firm ideals and beliefs.
Curriculum ; Humans ; Universities

Objective: To explore the clinical phenotype, immunological features, pathogenesis and gene variation of a case with A20 haploinsufficiency (HA20). Methods: A patient diagnosed with tumor necrosis factor α-induced protein 3 (TNFAIP3) mutated HA20 was admitted into Shenzhen Children′s Hospital in May,2019.The clinical data was analyzed. Flow cytometry was used to detect the patient′s peripheral blood lymphocyte subsets, and also, the percentage of follicular helper T cell (TFH) cells in the patient and thirteen healthy controls. After the construction of empty vector, wild-type and mutant plasmid vectors, a wild-type or mutant overexpression system of the TNFAIP3 gene was established in 293T cells and Hela cells. Then, the expression level of A20 in 293T cells and the expression of inhibitor K binding α (IKBα) in green fluorescent protein (GFP)+Hela cells before and after tumor necrosis factor α (TNF-α) stimulation were measured, to verify the pathogenicity of this variation. Results: A 5 years and 11 months old boy, presented with recurrent oral ulcer, abdominal pain, joint swelling and arthralgia. Oral ulcer, chronic skin rashes, knee joint swelling were observed. The levels of inflammatory markers were increased. Colonoscopy showed congestion of mucosa and multiple ulcers in terminal ileum and ileocecus. The absolute number of naive B cells was 124×10⁶ cells/L (reference range 147×10⁶-431×10⁶ cells/L), accounting for 0.430 of the total B cells (reference range 0.484-0.758). Compared to healthy controls (0.016-0.071), the percentage of TFH cells in CD4⁺T cells was much lower (0.008).A heterozygous mutation of TNFAIP3 gene (c.909_913 del, p.L303fs) was identified by genetic analysis. In vitro study showed that truncated A20 protein was expressed in TNFAIP3 mutant overexpressed 293T cells, which verified the pathogenicity of this variation. Besides, after TNF-α stimulation, the degradation rate of IkBα protein in mutant overexpressed Hela cells (35%) was between the other two groups (15% in the wild-type group and 57% in the non-loaded group). Conclusions This case with HA20 due to a de novo TNFAIP3 gene mutation presents with early onset Behcet-like autoinflammatory syndrome. This variation leads to expression of truncated A20 protein, enhanced degradation of IkBα, and further activation of nuclear factor κB signaling pathway.

Natural extracellular vesicles (EVs) play important roles in many life processes such as in the intermolecular transfer of substances and genetic information exchanges. Investigating the origins and working mechanisms of natural EVs may provide an understanding of life activities, especially regarding the occurrence and development of diseases. Additionally, due to their vesicular structure, EVs (in small molecules, nucleic acids, proteins, etc.) could act as efficient drug-delivery carriers. Herein, we describe the sources and biological functions of various EVs, summarize the roles of EVs in disease diagnosis and treatment, and review the application of EVs as drug-delivery carriers. We also assess the challenges and perspectives of EVs in biomedical applications.