Objective To evaluate cytotoxic effects of cytokine-induced killer cells (CIK cells) transfected with the interleukin-2 (IL-2) gene on malignant melanoma cells.Methods Mouse spleen cells were extracted,lymphocyte cells were separated,and CIK cells were prepared from these lymphocyte cells.PEGF-N1 plasmids containing IL-2 gene (PEGF-NI-IL-2) were transfected into CIK cells.Fluorescence microscopy was used to observe transfection products,and reverse transcriptase-polymerase chain reaction (RT-PCR) was conducted to determine the IL-2 mRNA expression.Then,effector cells such as CIK cells and IL-2-transfected CIK cells were separately co-cultured with target cells (B16 melanoma cells) at effector-target ratios of 10∶ 1,20∶1 and 40∶1,then 4-hour lactate dehydrogenase release assay was performed to evaluate cytotoxic effects of the two kinds of CIK cells on B 16 cells.After effector cells were cocultured with target cells at the effector-target ratio of 40∶1 for 48 hours,enzyme-linked immunosorbent assay (ELISA) was conducted to detect levels of IL-2,interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in the supernatant of the two kinds of CIK cells.Finally,mouse models of melanoma were established,and a total of 28 melanoma-bearing mice were divided into 4 groups to be peritumorally injected with 0.2 ml sodium chloride physiological solution (control group),100 IU IL-2 solution (IL-2 group),CIK cell suspension at a cell density of 1 × 106 cells per milliliter (CIK group) and IL-2-transfected CIK cell suspension at a cell density of 1 × 106 cells per milliliter (IL-2-transfected CIK group) respectively.Tumor morphology,tumor inhibition rate and cell apoptosis rate were used to evaluate tumor growth in the above groups.If data were normally distributed,t-test was used for comparing means between two groups,and analysis of variance and least significant difference (LSD)-t test were used for comparing means among multiple groups.Results Fluorescence microscopy and RT-PCR both showed that IL-2 was successfully transfected into CIK cells.The cytotoxic effect of IL-2-transfected CIK cells on B16 cells was strongest at the effector-target ratio of 40:1.Levels of IL-2,IFN-γ and TNF-α were also significantly higher in the supernatant of IL-2-transfected CIK cells [(1107.26 ± 6.49) pg/ml,(50.01 ± 3.35) pg/ml,(39.86 ± 3.25) pg/ml] than those in that of CIK cells [(51.09 ± 3.85) pg/ml,(32.71 ± 2.43) pg/ml,(30.11 ± 3.08) pg/ml,t =442.60,14.93,6.89,all P ＜ 0.01].Animal experiments showed that the tumor volume obviously increased in the control group (P ＜ 0.05),but significantly decreased in the IL-2 group,CIK group and IL-2-transfected CIK group (all P ＜ 0.001) after intervention compared with those before intervention.Furthermore,the tumor volume in the IL-2-transfected CIK group was significantly less than that in the other three groups (all P ＜ 0.01),but no significant difference was observed between the IL-2 group and CIK group (P ＞ 0.05).In addition,the apoptosis rate was significantly higher in the IL-2 group,CIK group,and IL-2-transfeeted CIK group than that in the control group (all P ＜ 0.01).The apoptosis rate and tumor inhibition rate were significantly higher in the IL-2-transfected CIK group than those in the IL-2 group and CIK group (all P ＜ 0.01),but insignificantly different between the IL-2 group and CIK group (P ＞ 0.05).Conclusion IL-2-transfected CIK cells had stronger killing effects on malignant melanoma.

Objective To expand the influence and promotion effect of the grassroots health demonstration base of appropriate technology at county level,explore the practice model for full coverage.Methods Four consortium and eight units in the county were engaged into the whole process,the whole cycle,synchronous implementation;the promotion practices were divided into different stages with different focuses based on priority setting;Stratified training,classified promotion strategies were involved to carry out the appropriate technology for all 11 items covered.Results The technical promotion training,technical promotion applications were completed with full coverage in the county,gained high satisfaction from both medical staff and public.Enhanced the technology radiation ability,also the base's annual development was increasing year by year.Conclusions The base construction full coverage promotion experiences can be shared and learned by other areas which aims for the promotion of fit health techniques.

Objective To provide effective reference for quality analysis of the chemical composition and extraction of astragalus separation process by comparing the extract of astragalus and it’s IR spectra. Methods The saponins and flavonoids in astragalus were firstly extracted by the method of circumfluence with ethanol as solvent and the residue of ethanol-extraction was then used to extract polysaccharides by distilled water. Fourier transform infrared spectroscopy (IR) combined with second derivative infrared spectroscopy was applied to quickly identify astragalus herbs powder, water extraction of astragalus, astragalus alcohol extraction and water extraction of the residue of ethanol-extraction. Results The powder and 70% ethanol extract, 80% ethanol extract were around at 1 735 cm-1 (carbonyl stretching vibration absorption peak) has a weak, broad absorption, while the absorption peak was less obvious in boiling water extraction. So the maln components of astragalus water extraction are polysaccharides and also contaln a small amount of water-insoluble flavonoids. Alcohol extraction malnly contalns saponins and flavonoids, and flavonoid extract volume increases with increasing alcohol concentration in a certaln range.Conclusion This method can be a quick and easy identification for astragalus and it’s extraction for its chemical composition class, and provide the basis for further research quality.

Objective:To explore the possible related factors of Beh?et's disease complicated with tendinitis, in order to better understand the etiology and development mechanism so to guide clinical diagnosis and treatment.Methods:The clinical data of patients with Beh?et's disease complicated with tendonitis treated at Department of Rheumatology and Immunology of Lanzhou University Second Hospital from October 2018 to September 2019 were retrospectively reviewed and related literature were reviewed.Results:Two patients were diagnosed as Beh?et's disease. Foot pain occurred during the treatment. Ultrasound showed tendonitis, and the corresponding treatment relieved the symptoms.Conclusion:Tendons may be involved and presents as a chronic change in patients with Beh?et's disease. In patients with rheumatic diseases, attention should be paid to the correlation between the disease and tendonitis. Aggressive treatment can prevent adverse consequences.

Objective To investigate the inhibitory effect of T ubacin,a selective inhibition of histone deacetylase 6 (HDAC6),on the release of inflammatory mediators in lipopolysaccharide (LPS) activated microglias and its underlying mechanism.Methods BV-2 microglias were divided into control group (conventional culture),LPS group (100 ng/mL LPS),Tubacin treatment group (1 μmol/L Tubacin) and experimental group (LPS 100 ng/mL+Tubacin 1 μmol/L).The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA).The production of nitric oxide (NO) was assayed by Griess reagent and the expression of inducible nitric oxide synthase (iNOS) was measured by Western blotting.The oxidative stress levels of BV-2 cells were determined by reactive oxygen species (ROS) and superoxide dismutase (SOD) assays.Results As compared with those in the control group,the productions of IL-6,TNF-α and NO were notably increased,the iNOS protein expression was significantly up-regulated,the ROS level was apparently elevated and the SOD activity was significantly decreased in the LPS group (P＜0.05).As compared with those in the LPS group,the productions of IL-6,TNF-α and NO were notably decreased,the iNOS protein expression was significantly down-regulated,the ROS level was apparently lessened and the SOD activity was significantly increased in the experiment group (P＜0.05).Conclusion Tubacin curbs the release of inflammatory mediators in activated microglial cells induced by LPS,whose effect may be achieved through decreasing oxidative stress levels in LPS activated microglial cells.