Objective:To investigate the clinical value of high intensity focused ultrasound (HIFU) ablation combined with levonorgestrel releasing in uterine system (LNG-IUS) in the treatment of adenomyosis.Methods:From January 2015 to January 2018, patients with adenomyosis diagnosed and treated in Obstetrics and Gynecology of our hospital were selected as the research objects. According to the wishes and treatment methods of the patients, the patients were divided into two groups: the combined treatment group was treated with HIFU combined with LNG-IUS, and the HIFU group was only treated with HIFU; the lesion volume and uterine volume reduction rate, hemoglobin, carbohydrate antigen 125 (CA125) and verbal rating scale (VRS) scores were compared between the two groups before and after treatment, and adverse reactions after treatment the scores were recorded.Results:Cases were followed up for 12 months after treatment: the lesion volume reduction rate and uterine volume reduction rate in the combined treatment group were better than those in the HIFU group [(58.8±14.1)% vs (49.3±17.2)%, (37.4±6.2)% vs (32.9±5.0)%, P<0.05]; the hemoglobin level was higher than that in the control group [(113.4±12.8)g/L vs (107.5±10.6)g/L, P<0.05]. In addition, the VRS score of dysmenorrhea in the combined treatment group was also better than that in the HIFU group, with statistically significant difference ( P<0.05). Conclusions:Compared with HIFU ablation alone, patients performed in HIFU combined with LNG-IUS achieved better lesion volume reduction rate and uterine volume reduction rate, and hemoglobin level and dysmenorrhea symptoms improved better in patients with adenomyosis.

Objective: To investigate the anti-tumor effect of CTL cells on colon cancer xenograft in nude mice after knocking out the immune check point CTLA-4 by CRISPR/Cas9 technology. Methods: A specific small guide RNA (sgRNA) for CTLA-4 was designed to construct sgRNA/Cas9 plasmid, which was then transfected into CTL using a lentiviral vector to obtain CTL cells with CTLA-4 deletion (CTLA-4 KO CTL). The transfection efficiency of the plasmid and the deletion efficiency of CTLA-4 were verified. BALB/c nude mice were randomly divided into two groups to prophylactically inoculate CTLA-4 KO CTL (experimental group) or CTL (control group); 3 days later, the animals of two groups were inoculated with colon cancer cell line LS174-T to observe the tumor formation rate and tumor formation time. After constructing colon cancer xenograft model in nude mice, the animals were randomly divided into two groups, respectively treated with CTLA-4 KO CTL (experimental group) and CTL (control group) cells to observe the tumor growth volume and survival time of mice. The serum levels of TNF-α and IFN-γ in nude mice were detected. Results: sgRNAwas designed and CRSIPR/Cas9 system with lentivirus as vector was successfully constructed. CTL cells were transfected with the established CRSIPR/ Cas9 system, and the highest transfection efficiency was up to (28.80±0.62)%. After transfection, the deletion efficiency of CTLA-4 was detected by Flow cytometry. The CTLA-4 expression of CTLA-4 KO CTL group was significantly lower than that of CTL group [(0.91±0.25)% vs (42.70±2.72)%, P<0.05]. In prophylactic assay, the formation rate of colon cancer xenografts in the experimental group was significantly lower than that in the control group(33.33%vs100%,P<0.05). In treatment assay, the tumor volume in the experimental group was significantly inhibited compared with the control group ([503±23.9] vs [911.2±51.4] mm3, P<0.05), and the survivaltimeoftheexperimentalgroupwassignificantlyprolonged (mediansurvivaltime:78dvs42d,P<0.05); Moreover, the secretion levels of serumTNF-α([268.93±17.04]pg/mlvs[148.26±20.07]pg/ml,P<0.05) and IFN-γ(315.38±18.67 pg/ml vs 202.92±29.32 pg/ml, P<0.05) in the experimental group were significantly higher than those in the control group. Conclusions: The lentiviral vector CRSIPR/Cas9 system is an effective gene editing method; its successful deletion of CTLA-4 in CTL cells can significantly inhibit the tumor formation rate of colon cancer xenografts in nude mice and enhance the anti-tumor effect of CTLon colon cancer xenografts.