Objective To investigate the changes of regulatory T cells (Treg) in patients with hepatocellular carcinoma (HCC) after ultrasound- guided percutaneous cool- tip radiofrequency ablation (RFA), and to discuss its influence on the prognosis. Methods A total of 30 patients with HCC were enrolled in this study. The percentage of Treg in peripheral blood was estimated with flow cytometry before RFA and one, 4, 7 and 12 months after RFA. During the follow-up period, the therapeutic effects were evaluated by contrast enhanced sonography or contrast enhanced CT scanning. By using the methods of receiver operating characteristic (ROC) curve and Kaplan-Meier survival function, the correlation of Treg dynamic changes with the progression-free survival time was analyzed. Results One month after RTA, the tumor response (TR) rate in the 30 patients was 93.3% (28/30), the tumor progression (TP) rate was 6.67%(2/30). The percentage of Treg before RFA was (9.42 ± 1.16)%, which decreased to (6.55 ± 0.97)% one month after RFA, the difference was statistically significant (t = 15.325, P <0.001). Twelve months after RFA, TR rate became 33.3%(10/30), and TP rate became 66.7%(20/30). The preoperative percentage of Treg of TR group was (8.75 ± 0.72)%, which was significantly lower than that of TP group (9.76 ± 1.20)%, the difference was statistically significant (t=-2.448, P=0.021). ROC curves indicated that the optimal cut-off value of Treg nadir was 4.82%, the sensitivity was 90.0% and the specificity was 60.0%. The optimal cut-off time to reach Treg nadir was 5.5 months, the sensitivity was 70.0% and the specificity was 85.0%. Kaplan-Meier curve analysis showed that after RFA the progression-free survival rate (PFS) of patients with Treg nadir ≤ 4.82% was significantly higher than that of patients with Treg nadir>4.82%. PFS of patients with reaching Treg nadir≥5.5 months was significantly higher than that of patients with reaching Treg nadir<5.5 months. Log-rank test results were字2=5.207, P=0.023; 字2=22.079, P < 0.001, respectively. Conclusion Percutaneous cool-tip radiofrequency ablation can decrease the percentage of Treg cells. Besides, Treg nadir and the time reaching Treg nadir can reflect the prognosis of HCC patients after RFA to a certain extent.

Objective: To investigate the effect and safety of apatinib in the treatment of advanced gastric cancer. Methods: A total of 60 patients with advanced gastric cancer at the first Affiliated Hospital of Hainan Medical College from April 2015 to October 2018 were retrospectively analyzed, and all patients were divided into two groups according to the treatment method, each group of 30 cases. The control group was treated with conventional treatment, and the observation group was given the treatment of apatinib on the basis of the control group, and then the clinical curative effect and adverse reactions were compared between the two groups. Results: There were 2 cases (6.67%) of complete remission in the observation group, 14 cases (46.67%) of partial remission, 10 cases (33.33%) of stability and 4 cases (13.33%) of disease progression, and the total effective rate was 53.33% (16/30). The control group had no complete remission, 8 cases (26.67%) of partial remission, 11 cases (36.67%) of stability and 11 cases (36.67%) of disease progression, and the total effective rate was 26.67% (8/30). The difference of total effective rate between the two groups was statistically significant (χ ² = 4.444, P = 0.035). There was no significant difference in the level of tumor marker carcinoembryonic antigen (CEA), carbohydrate antigen (CA)199, CA125, cytokeratin 19 fragment antigen 21-1 (CYFRA21-1) between the two groups before and after treatment (all P > 0.05). The incidence of hand-foot syndrome and hypertension in the observation group was higher than that in the control group [73.33% (22/30) vs. 43.33% (13/30), H = 5.554, P = 0.018; 76.67% (23/30) vs. 46.67% (14/30), H = 5.711, P = 0.017]. The incidence of proteinuria, bone marrow suppression and the cardiac adverse reactions in the observation group was higher than that in the control group [63.33% (19/30) vs. 33.33% (10/30), H = 3.300, P = 0.069; 60.00%(18/30) vs. 36.67% (11/30), H = 5.455, P = 0.071; 53.33% (16/30) vs. 30.00% (9/30), H = 3.360, P = 0.067]. Conclusion Apatinib is effective in the treatment of advanced gastric cancer, but the occurrence of hand-foot syndrome and hypertension should be paid close attention during the treatment.

Objective To compare the efficacy of erlotinib and pemetrexed in treatment of advanced non-small cell lung cancer (NSCLC) and its effect on the immunity of patients. Methods A total of 82 patients with NSCLC at the First Affiliated Hospital of Hainan Medical College from June 2014 to May 2018 were retrospectively analyzed. According to the different treatment methods, the patients were divided into erlotinib group (150 mg/d, oral administration, 2 hours after the meal) and pemetrexed group (500 mg/m2, intravenous drip, 21-day each cycle). There were 41 cases in each group. The clinical effects of the two groups were analyzed. Flow cytometry was used to detect the immune index. Results The objective effective rate in pemetrexed group was lower than that in erlotinib group [34.2％ (14/41) vs. 39.0％ (16/41), χ 2 = 0.210, P =0.647). There was no significant difference in T lymphocyte subsets between the two groups before and after treatment (both P > 0.05). The values of CD3+ T cells, CD4+ T cells, CD4+ / CD8+ in both groups after the treatment were decreased compared with the values before the treatment; CD8 + T cells was increased after the treatment compared with the value before the treatment (all P < 0.05). Quality of life questionnaire-C30 (QLQ-C30) score in erlotinib group was higher than that in pemetrexed group [(75.1±13.5) vs. (68.9±12.9), t = 2.158, P = 0.017]. Myelosuppression and gastrointestinal reactions were the main adverse events in pemetrexed group; rashes and diarrhoea were the main adverse reactions in erlotinib group. Grade Ⅰ-Ⅱ side effects occurred in both groups. There were statistical differences in the incidence of myelosuppression, gastrointestinal reactions, rashes and diarrhoea of both groups (all P < 0.05). Conclusions The efficacy oferlotinib and pemetrexed in treatment of advanced NSCLC is similar, and both of them could regulate the immune response. Erlotinib has a significant advantage in improving the quality of life.

Objective The aim of this study was to investigate the synergistic anti-tumor effect of cryptotanshinone and cis-platin in non-small cell lung cancer NCI-H1975 cells and its possible molecular mechanism. Methods NCI-H1975 cells were treated with control,cryptotanshinone,cisplatin or combination of cryptotanshinone and cisplatin groups(referred to as the combination group). The inhibitory rate of cell proliferation was determined in NCI-H1975 cells by CCK-8 assay;Flow cytometry was used to determine the rates of survival and cell apoptosis;The expression of apoptotic protein,anti-apoptotic protein,JAK2/STAT3 protein and its phosphorylation levels were detected in NCI-H1975 cells by Western blot. The localization and transcription activity of STAT3 cells were determined by laser confocal microscopy/luciferase assays. Results 1. The survival rates in cryptotanshinone,cisplatin and combination groups at each time-point were lower in NCI-H1975 cells than that in the control group(P<0. 05);The proliferation of NCI-H1975 cells in the combination group were inhibited when compared to the control,cryptotanshinone,or cisplatin groups(P<0. 05);2. After treatments for 24 h,the expression levels of Bcl-2 and Survivin protein were decreased(P<0. 01),the activity of Caspase3 and Caspase9 protein was increased(P<0. 01),and the expression levels of p-STAT3 and p-JAK2 protein were de-creased in NCI-H1975 cells(P<0. 01);3. After NCI-H1975 cells treated with the combination of cryptotanshinone and cisplatin, the STAT3 in the nucleus was decreased,and STAT3 activity was also decreased in the nucleus. Conclusion Cryptotanshinone com-bined with cisplatin can exert a synergistic antitumor effect on non-small cell lung cancer NCI-H1975 cells,and its mechanism is related to the inhibition of JAK2 and STAT3 phosphorylation signal pathways.

Objective To investigate the differential expression profile of epithelial mesenchymal transition (EMT) related long chain noncoding RNA (LncRNA) in hepatocellular carcinoma (HCC) cells after incomplete radiofrequency ablation (RFA) in vitro.Methods Incomplete RFA was simnulated in vitro by using Huh7 cells in water bath at 47℃.EMT change was detected by microscopy and Western blot.Cell invasion and migration were detected by transwell assays.Cell proliferation was determined by cell counting kit-8 (CCK-8) assay.Differential expression profile of EMT-related LncRNAs between Huh7-H and Huh7 were analyzed by LncPath human EMT array,and it was validated by RT-PCR.Results Under microscopy,Huh7-H presented characteristic EMT morphological changes.Western blot analysis showed that the expression of E-cadherin in Huh7-H cells was decreased,while the expressions of N-cadherin and Vimentin were increased.Transwell assay test indicated that cell migration and invasion were increased in Huh7-H cell when compared with Huh7 cell (61.0±5.2 vs 138.0±11.8 and 33.3±7.8 vs 82.7±39.4,respectively),the abilities of Huh7-h cell in migration and invasion were evidently strengthened (P＜0.05).CCK-8 assay shawed that the proliferation ability of Huh 7-H was obviously higher than that Huh 7 (P＜0.05).LncPath human EMT array screened out 3 differential expressed LncRNAs (P≤0.05 and fold changes ≥ 1.5),including two down-regulated LncRNAs (FUNDC2P4,RPL27P7) and one up-regulated LncRNA (MTND4LP14).RT-PCR validation revealed that the expressions of FUNDC2P4,RPL27P7 and MTND4LP14 in Huh7-H cells were 0.137,0.869 and 1.037times of that in Huh7 cells,respectively.Clinical specimen validation showed that the expression of LncRNA FUNDC2P4 in peficancerous tissue was higher than that in HCC tissue,and the expression of LncRNA FUNDC2P4 in HCC tissue was higher than that in the residual HCC tissue after RFA.Conclusion EMTrelated LncRNA FUNDC2P4 in HCC cells with low expression after incomplete RFA is successfully screened out,which provides basis for the further investigation of the function and molecular mechanism of this gene.