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@#Objective: To prepare human peripheral blood T lymphocytes with TIM3 (T cell immunologlobulin and mucin-3) gene knockout by using CRISPR-Cas9 gene editing technique and plasmid electrotransfection system, and to discuss whether the knockout of TIM3 gene could enhance the immune response and anti-tumor efficacy of T cells. Methods: Double plasmids hTIM3 sgRNA/Cas9 were transfected into human peripheral blood T lymphocytes of EBV positive gastric cancer patients by using electrotransfection system. The transfection efficiency was examined 24 h later by flow cytometry and fluorescence microscope. The proliferation activity of the T cells after gene knockout was observed during in vitro culture, and the knockout efficiency and phenotypes of the modified T cells were evaluated by flow cytometry. Furthermore, tumor antigen peptide was used to activate T cells, and the level of modified T cells secreting cytokines and its cytotoxicity against gastric cancer AGS-EBV cells were evaluated. Results: Electrotransfection system could successfully transfect hTIM3 sgRNA/Cas9 double plasmids into human peripheral blood T lymphocytes with an average transfection efficiency of (41.5±3.6)%, and the gene knockout efficiency fluctuated between 40.0% and 50.0% (all P<0.01). The proliferation of the modified T cells was not significantly changed in the TIM3 gene knockout group even after the prolonged co-culturing with tumor antigenic peptide;and for the activated molecules, only HLA-DR exhibited significant elevation as compared with control group (P<0.05). Remarkably, T cells with TIM3 gene knockout showed significantly elevated secretion of TNF-α and IFN-γ (P<0.01 or P<0.05), as well as obviously enhanced in vitro cytotoxicity against gastric cancer AGS-EBV cells (P<0.05). Conclusion: It’s simple and feasible of CRISPR-Cas9 gene editing technique and plasmid electrotransfection system to prepare T lymphocytes with engineered TIM3 gene knockout. The expression level of TIM3 was down-regulated in in vitro culture. More importantly, the modified T cells performed superior immune response and cytotoxicity, which may provide a new idea for gene engineering cell immunotherapy.
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Objective To understand the prevalence of synthetic drug use among man who have sex with man (MSM) in Nanjing and the associated factors. Methods MSM were recruited by snow-ball sampling, peer recommendation and network recruitment. Multivariate logistic regression analysis was performed to evaluate the correlated factors of recreational drug use. Results 29.6% of the 876 MSM ever used recreational drugs. The prevalence of HIV and syphilis infections were 9.5% and 9.0% respectively. After controlling the influence of correlated factors, recreational drug use was correlated with unprotected anal intercourse in the past 6 months(OR=1.83, 95% CI: 1.35-2.47, P<0.001), multiple male sex partnership in the past 6 months(OR=2.25, 95% CI: 1.65-3.05, P<0.001), higher HIV prevalence (OR=1.95, 95% CI: 1.21-3.17, P=0.007) and higher syphilis prevalence(OR=1.71, 95% CI: 1.02-2.84, P=0.040). Multivariate Logistic analysis showed that demographic and behavioral factors associated with recreational drug use including: less than 30 years old (OR=1.50, 95% CI: 1.07-2.11, P=0.020), self-identified as homosexual orientation (OR=1.65, 95% CI: 1.16-2.34, P=0.005), seeking male partners mainly via the internet(OR=1.58, 95% CI: 1.07-2.35, P=0.022), higher monthly income(OR=1.57, 95% CI: 1.13-2.17, P=0.007), receptive sexual role(OR=1.55, 95% CI: 1.04-2.32, P=0.033). Conclusions MSM has a high prevalence of recreational drug use in Nanjing, which was positively correlated with HIV/syphilis infection and high risk behaviors. Intervention efforts should be devoted to reduce and combat drug use among MSM.
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Objective To investigate the effect of topiramate in the treatment of intractable epilepsy in children. Methods 120 cases with intractable epilepsy from May 2012 to May 2017 were randomly divided into 2 groups, the control group used antiepileptic drug treatment, study group combined application of anti epileptic drugs targeted nursing application. Results The clinical effect of the treatment group was better than that of the control group (P< 0.05). The frequency of epileptic seizure in the treatment group was significantly higher than that in the control group (P<0.05). Conclusion In the treatment of intractable epilepsy in children, the application of topiramate in the treatment of basic care is helpful to improve the effect of treatment, the effect is ideal, and should be further popularized in clinical practice.
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Objective To investigate the effect of topiramate in the treatment of intractable epilepsy in children. Methods 120 cases with intractable epilepsy from May 2012 to May 2017 were randomly divided into 2 groups, the control group used antiepileptic drug treatment, study group combined application of anti epileptic drugs targeted nursing application. Results The clinical effect of the treatment group was better than that of the control group (P< 0.05). The frequency of epileptic seizure in the treatment group was significantly higher than that in the control group (P<0.05). Conclusion In the treatment of intractable epilepsy in children, the application of topiramate in the treatment of basic care is helpful to improve the effect of treatment, the effect is ideal, and should be further popularized in clinical practice.
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<p><b>OBJECTIVE</b>To assess the value of contrast-enhanced ultrasound imaging in monitoring the therapeutic effect of argon-helium cryosurgical treatment of malignant tumors.</p><p><b>METHODS</b>Before and after argon-helium cryosurgical treatment, 42 patients underwent contrast-enhanced ultrasound imaging, conventional ultrasound imaging and enhanced CT or magnetic resonance imaging (MRI) for examining the number of tumor foci and the size of necrotic areas.</p><p><b>RESULTS</b>A total of 80 tumor lesions were detected by contrast-enhanced ultrasound imaging. Compared with conventional ultrasound imaging, contrast-enhanced ultrasound imaging detected a significantly greater number of tumors and the intratumoral necrotic areas (96 vs 19) as well as a significantly increased mean size of necrotic areas (5.7∓3.6 cm vs 2.8∓1.7 cm), showing no significant differences from the results by enhanced CT and MRI (94 and 5.5∓3.3 cm, P=0.872 and 0.978, respectively). The short-term therapeutic effect of argon-helium cryosurgery evaluated by contrast-enhanced ultrasound imaging were also similar to that assessed by enhanced CT or MRI (P=0.906).</p><p><b>CONCLUSION</b>Contrast-enhanced ultrasound imaging has important values in monitoring malignant tumors during argon-helium cryosurgical treatment and in evaluating the short-term therapeutic effect of the treatment.</p>