ABSTRACT
Objective@#To establish a method for in vitro expansion of regulatory T cells (Tregs) for clinical study and immunotherapy.@*Methods@#CD4+ T cells were isolated from BALB/c spleens by magnetic activated cell sorting (MACS), then cultured in 24-well plates coated with anti-CD3/CD28 microbeads in the presence of 100 U/ml interleukin-2 (IL-2) and 5 ng/ml recombinant human transforming growth factor-β (rhTGF-β). The purity of the expanded Tregs were tested by flow cytometry. In vitro inhibition of CD4+ CD25-T cells response by expanded Tregs was measured by mixed lymphocyte reaction test. The number and the viability of the expanded Tregs were detemined by trypan blue staining.@*Results@#Tregs were expanded up to 20 folds after 5 days in culture, and the activity was (93±4)%. The purity of expanded Tregs were (79.1±1.5)%, and maintained suppressive ability.@*Conclusions@#We can obtain large numbers of Tregs with highly purity and suppressive ability, thereby providing a solution to the availability of sufficient Tregs in clinical study and immunotherapy.
ABSTRACT
Objective To establish a method for in vitro expansion of regulatory T cells (Tregs)for clinical study and immunotherapy.Methods CD4 + T cells were isolated from BALB/c spleens by magnetic activated cell sorting (MACS),then cultured in 24-well plates coated with anti-CD3/CD28 microbeads in the presence of 100 U/ml interleukin-2 (IL-2) and 5 ng/ml recombinant human transforming growth factor-β (rhTGF-β).The purity of the expanded Tregs were tested by flow cytometry.In vitro inhibition of CD4 + CD25-T cells response by expanded Tregs was measured by mixed lymphocyte reaction test.The number and the viability of the expanded Tregs were detemined by trypan blue staining.Results Tregs were expanded up to 20 folds after 5 days in culture,and the activity was (93 ± 4) ％.The purity of expanded Tregs were (79.1 ± 1.5) ％,and maintained suppressive ability.Conclusions We can obtain large numbers of Tregs with highly purity and suppressive ability,thereby providing a solution to the availability of sufficient Tregs in clinical study and immunotherapy.
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Objective To investigate the efficacy and safety of early goal-directed sedation with dexme-detomidine in patients with septic shock.Methods This study is selected from August 2015 to December 2016 in our department 70 patients receiving treatment of septic shock,and through random sample table method divided into the control group and the study group two,there were 32 cases in control group,the control group selected ob-ject according to the conventional sedation treatment,namely the application of midazolam extraction treatment, the study group of 38 patients with septic shock in patients with application of dexmedetomidine sedation,analysis and application of different sedation method analysis of two groups of patients with septic shock. The efficacy and safety of observation and etc. Results Two groups of patients with different treatment methods,The onset time of the study group of patients with septic shock after sedation,h recovery time and time of mechanical ventilation was obviously better than the control group,the effect of difference(P < 0.05),has significant research value. Conclusions For patients with septic shock early goal orientation selection and application of sedation of dexme-detomidine can play a better effect,the more safe and effective,sedation score is high,it is worth of application in clinical treatment.
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Objective To explore the clinical effect of acupuncture on acute renal injury of sepsis.Methods From January 2015 to May 2017,adult patients with sepsis who received ≥ 7 days of treatment were collected from the Central Hospital of Zhuzhou.The participants were randomly divided into the control group (n =35) and the treatment group (n =37),and the control group was given routine treatment according to the sepsis guidelines.On the basis of control group,the treatment group was given acupuncture treatment,acupoints Shenshu and Sanyinjiao,Taixi,Zusanli.The clinical effects of the two groups of patients were compared.Results The levels of cystatin C,blood urea nitrogen and creatinine in the treatment group were significantly lower than those in the control group (P ＜ 0.05).There was no significant difference in the number and proportion of patients with blood purification between the two groups (P ＞ 0.05),while the frequency and time of blood purification treatment in the treatment group were significantly lower than those in the control group (P ＜ 0.05).The levels of blood interleukin-6 and tumor necrosis factor-alpha in the treatment group were significantly lower than those in the control group (P ＜ 0.05).From the fourth day of treatment,the Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score in the treatment group was significantly lower than that in the control group (P ＜ 0.05).There was no significant difference in mortality between the two groups at 28 days (P ＞ 0.05).Conclusions Acupuncture of Shenshu,Sanyinjiao,Taixi,Zusanli can improve sepsis patients with acute kidney injury in renal function.