[Effectiveness of TRB3 on human hepatocellular carcinoma cells proliferation, apoptosis and migration].

Objective: To explore the regulatory role and mechanism of tribbles pseudokinase 3 (TRB3) on hepatocarcinoma (HCC) cells proliferation, apoptosis and migration. Methods: Immunohistochemistry and Western blot were used to detect TRB3 expression in cancerous and adjacent cancerous liver tissues of HCC patients. TRB3 expression was detected in vitro in HepG2 and Huh7 hepatocarcinoma cell lines. Simultaneously, CCK8 and EdU were used to detect cell proliferation after TRB3 targeted inhibition with small interfering RNA. CCK8 and EdU were used to detect cell proliferation. Flow cytometry assay was used to detect apoptosis. Transwell assay was used to evaluate migration ability. Simultaneously, Western blot was used to detect changes in apoptosis, migration-related proteins and AKT phosphorylation activity. The mean comparison between the two groups was performed by t-test, and the comparison between multiple groups was performed by one-way analysis of variance. Results: Western blot showed that the expression of TRB3 was significantly up-regulated in HCC tissues. Compared with normal liver tissues adjacent to cancer, the relative expression levels were 0.78 ± 0.12 and 0.29 ± 0.09, respectively, P < 0.01, and the difference was statistically significant. After interfering siRNA inhibited TRB3, CCK8 and EdU tests showed that the proliferation activity of HepG2 and Huh7 cells were significantly weakened (P < 0.05). Flow cytometry results showed that the apoptotic proportions of HepG2 and Huh7 cells was significantly increased (P < 0.01). Western blot also showed that the expression of apoptosis regulatory proteins BAX and BIM were significantly increased (P < 0.01). Transwell assay results showed that the migration ability of HepG2 and Huh7 cells was decreased (P < 0.05), and the expression of migration regulatory proteins MMP4 and MMP9 was also significantly down-regulated. Western blot results showed that the AKT phosphorylation level was significantly increased. Conclusion: TRB3 regulates hepatocarcinoma cells proliferation, apoptosis and migration by inhibiting the AKT phosphorylation activity. Therefore, TRB3 may be a potential target site for the liver cancer treatment.

LncRNA DCST1-AS1 accelerates the proliferation, metastasis and autophagy of hepatocellular carcinoma cell by AKT/mTOR signaling pathways.

OBJECTIVE: The previous research revealed that long noncoding RNAs (lncRNAs) play a vital role in the development of hepatocellular carcinoma (HCC). To further discuss the underlying mechanisms of lncRNA DCST1-AS1 in the pathogenesis of HCC. PATIENTS AND METHODS: We screened the abnormally expressed genes in HCC tissues through microarray analysis and found that lncRNA DCST1-AS1 was one of the genes significantly up-regulated. Real Time-Polymerase Chain Reaction (RT-qPCR) was used to test the gene expression of lncRNA DCST1-AS1 in HCC tissues and HepG2 cells. Respectively, CCK-8 assay, flow cytometry detection, transwell assay, wound healing assay, transmission electron microscopy, and immunofluorescence staining were used to assess the proliferation, apoptosis, migration, and autophagy of HepG2 cells. Meanwhile, the expression of signaling pathway proteins was detected by Western blot. RESULTS: LncRNA DCST1-AS1 was confirmed hyper-expression both in HCC tissues and HCC cells. High expression of lncRNA DCST1-AS1 was significantly correlated with inferior prognosis. Moreover, lncRNA DCST1-AS1 depletion suppressed proliferation and accelerated apoptosis, activated cycle arrest, restrained cell migration, and stimulated autophagy in HCC cells. In addition, it is found that the depletion of lncRNA DCST1-AS1 on HepG2 cells exhibits anti-tumor characteristics and was mediated by the AKT/mTOR signal transduction pathway. Furthermore, pre-treated HepG2 cells with SC79, an AKT signal activator, partially restored the effect of lncRNA DCST1-AS1 silencing on HepG2 cell proliferation, apoptosis, migration, and autophagy. CONCLUSIONS: Our results suggested that lncRNA DCST1-AS1, as a carcinogenic factor in HCC, promoted cell proliferation, and invasion, inhibited apoptosis and autophagy by modulating the AKT/mTOR signaling cascade. Therefore, our findings showed that lncRNA DCST1-AS1 may improve potential treatment strategies for HCC.

[Comparative study on clinical efficacy of two surgical methods for gastric gastrointestinal stromal tumors at unfavorable position].

Objective: To investigate the safety and feasibility of laparoscopic operation in thetreatment of gastric gastrointestinal stromal tumor (GIST) at unfavorable positions. Methods: A retrospective cohort study was conducted to analyze the clinical data of patients with gastric GIST at unfavorable positions confirmed by pathology after surgery (laparoscopy or laparotomy) at the Southwest Hospital of the Army Medical University and the Minda Hospital of Hubei Minzu University from June 2008 to June 2018. The unfavorable positions of stomach are defined as the esophagogastric junction, the proximal cardia of gastric lesser curvature, the pylorus of stomach, the posterior wall of stomach and the antrum of stomach.Exclusion criteria:(1) preoperative chemotherapy or imatinib therapy; (2) diameter of tumor > 10 cm; (3) tumor metastasis or concurrence of other malignant tumors. A total of 244 patients (238 in Southwest Hospital and 6 in Minda Hospital) were enrolled, including 122 males and 122 females with age of 41-70years. Operative methods should be adopted according to patients' wishes. There were 146 cases in the laparoscopic surgery group and98 cases in the open surgery group. The intraoperative blood loss, operative time, postoperative first flatus time, postoperative firstfeeding time,average hospital stay, morbidity of postoperative complication,1-,3-,and 5-year recurrence free survival(RFS) and overall survival (OS)rate were compared between the two groups. Results: There were no significant differences in sex, age, tumor size, tumor risk grade or growth pattern between the laparoscopic and the open surgery groups (all P>0.05),and these two groups were comparable. Compared with open group, laparoscopic group had less intraoperative blood loss [(31.4±2.3) ml vs. (143.9±3.7) ml, t=292.800, P<0.001], shorter postoperative first flatus time [(2.1±0.7) days vs.(3.8±0.8) days, t=17.550,P<0.001], shorter postoperative first feeding time [(2.1±0.5) days vs.(2.3±1.7) days, t=1.339,P=0.020], shorter hospital stay [(8.6±2.6) days vs. (13.6±3.2) days, t=13.410, P<0.001], and lower morbidity of postoperative complication [16(11.0%) vs. 21(21.4%),χ2=4.996,P=0.025], whose differences were statistically significant. While the operation time was similar in two groups [(124.7±15.8) minutes vs. (120.9±14.5) minutes, t=1.903,P=0.058]. The median follow-up of all the patients was 43 (1 to 119) months.In laparoscopic group and open group, the 1-, 3- and 5-year RFS were 94.5% vs. 93.9%, 91.1% vs. 90.8%,and 82.2% vs. 81.6%, respectively, and 1-, 3- and 5-year OS were 98.6% vs. 95.9%, 95.9% vs. 94.9%,and 91.1% vs. 88.8%, respectively, whose differences were not statistically significant (all P>0.05). Conclusion: In the experienced gastrointestinal surgery center, laparoscopic resection of gastric GIST at unfavorable position is safe and feasible, achieving the same efficacy of open surgery.

Separation and expansion of tumor infiltrating lymphocytes of digestive system in vitro and their cytotoxicity.

Primary tumor tissues in the digestive system were harvested from 15 patients. By mincing, enzymatic digestion and gradient density separation, sufficient TILs (> x 10(6)) were obtained from 13 of 15 (88.7%) patients in vitro in the presence of 500 mu/ml of recombinant interleukin-2 and 5% fetal calf serum after one month culture. 92.3% (12/13) of TILs proliferated well in vitro (92.3%). TILs expanded from 10(/)-folds to 10(3)-folds after being cultured for one month. CD25+ cell of the most fresh TILs was more than that of peripheral blood lymphocytes. CD25+ cells of TILs during 4th week of the culture was significantly greater (P < 0.01) than that of fresh TILs. CD4+/CD8+ ratio was decreased during four culture weeks because of increase of CD8 cells. By using modified colorimetric MTT assay for measuring activity of TILs against various tumor cells the results showed that cytotoxicity of gastro-intestinal TILs in autologous tumor cells is greater than on the other tumor cells.

Effect of the vertical force component of Class II elastics on the anterior intrusive force of maxillary archwire.

Class II elastics are usually employed in the treatment of excessive overbite and overjet with the Begg technique. The effect of Class II elastics on bite opening and the extent of such an effect is uncertain. This article, based on measurements of 30 cases, illustrates that the effect of the vertical component of force from Class II elastics in reducing the intrusive force generated by the anchor bands in the upper archwire is less than previously believed. The position of the circles in the archwire for intermaxillary elastics has the greatest influence on the anterior intrusive force of the upper archwire. The direction of the Class II elastics and the length of the dental arch (a negative correlation) came second, while the force actually exerted by the elastics had the smallest influence. It is, therefore, suggested that the position of the intermaxillary elastic circles should be located according to the differing clinical objectives.

[Trephine craniotomy in neurosurgery].

For three years, we have performed craniotomy with a 5 cm diameter trephine through a linear incision. 53 craniotomies were done in 49 patients. This simple and time-saving method produced little bleeding and slight injury to the brain. The technique allowed the surgeon to spend most of his time at the management of lesion. Healing of wound was rapid and brain edema was minimal.