Objective· To identify the effect and potential mechanism of gambogic acid (GA) on natural killer/T-cell lymphoma (NK/TCL) cell lines.Methods · SNK-1,SNK-6 and SNT-8 were incubated with various concentrations of GA for 24 h,and cell viability was detected with CCK-8 assay.Cell apoptosis was examined by Annexin V-FITC/PI staining assay.Levels of proteins regulating cell apoptosis and phosphorylation levels of proteins in key signaling pathways were detected by Western blotting.Results · GA showed a potent effect on reduction of cell viability of NK/fCL cell lines in CCK-8 assay.GA increased the percentages of Annexin V positive cells and induced activation of caspase-3 and caspase-9,cleavage of PARP as well as the reduction of Bcl-xl.GA also inhibited the phosphorylation levels of STAT3 in SNK-1 and SNT-8,and ERK1/2 in SNK-1 and SNK-6 significantly.Conclusion· GA induces cell apoptosis in NK/TCL cell lines SNK-1,SNK-6 and SNT-8.Anti-apoptosis protein Bcl-xl and signaling pathway JAKs/STATs and MEK/MAPK might be involved in this process.

Objective To investigate the method of establishing a model of delay eyeblink condi-tioning ( DEC) in no surgical cynomolgus monkey and analyze the related acquisition rule.Methods Using the special monkey chairs to fix 6 adult male cynomolgus monkeys,they were sitting on the chairs and keep-ing awake,and heads could rotate freely during the whole training session.In the experiment,the 6 monkeys were trained in DEC with a tone (500 ms,85 dB) as the conditioned stimulus (CS),paired with a corneal oxygen-puff (100 ms,5 psi) as the unconditioned stimulus (US).There were 120 trials per-session,and 2 sessions per-day.During the experiments,an infrared emitter/detector attached to spectacles for eyeblink re-cording,the data of conditioned response ( CR) and startle response ( SR) were analyzed offline.Result-s Among the 6 cynomolgus monkeys,4 of them completed all of the training process of DEC and 3 monkeys had successful acquisition of DEC.The average CR rate in the last training session reached ( 64.67 ± 2.00)%,(P<0.01);1 monkey only showed a high acquisition rate( 85.00%) at the first training session, and the low CR rate in the next training sessions with the average rate of 18.16%,suggesting that the DEC model was failed to established.The SR rates in 4 cynomolgus monkeys were low with average rate of 5.47%. Conclusion Although the DEC behavior training of the cynomolgus monkey is easy to be influenced by many inside or outside factors,DEC can be obtained under the condition of non surgery and proper braking.

Objective To explore the expression of miRNA-181a (miR-181a) in patients with multiple myeloma (MM) and its effect on biological features of MM cells. Methods CD138+cells of bone marrow from 25 MM patients and 10 patients with hematological non-malignancies were purified by using immunomagnetic separation, and the expression of miR-181a in CD138+cells and MM cell lines including RPMI 8226, H929 and U266 were detected by real-time quantitative PCR. The effects of down-regulation and up-regulation of miR-181a expression on the biological characteristics of MM cells were studied with miR-181a antagomir and agomir. Results Compared with patients with hematological non-malignant diseases, the expression of miR-181a in CD138+ cells was upregulated in MM patients. Compared with CD138+ cells in hematological non-malignancies, high expressions of miR-181a were observed in RPMI 8226 and U266 myeloma cell line, while low expressions of miR-181a were observed in H929 cells. Down-regulation of miR-181a with 100 nmol/L miR-181a antagomir could inhibit the proliferation of U266 cells at 24,48 and 72 h [(67.1 ± 3.3) %vs. (50.5 ± 4.1) %, (71.5 ± 3.6) % vs. (52.3 ± 2.2) %, (78.1 ± 5.4) % vs. (69.5 ± 4.3) %, P < 0.05 respectively], whereas up-regulation of miR-181a with 100 nmol/L miR-181a agomir could significantly promote the proliferation of H929 cells at 24 h and 48 h [(21.2 ± 2.4) %vs. (38.5 ± 3.6) %, ( 61.3 ± 5.4) %vs. (82.2 ±6.9)%, P<0.01 respectively]. Cell cycle analysis showed that miR-181a antagomir made U266 cell cycle arrest in the G0/G1 phase. Meanwhile, susceptibility test results indicated that the apoptosis of U266 cells induced by doxorubicin, paclitaxel and 5-fluorouracil was increased when the proliferation of miR-181a expression was down-regulated with miR-181a antagomir. In migration assay, the data showed that down-regulation of miR-181a with miR-181a antagomir could inhibit the migration of U266 cells, and the proportion of migrated cells in the experimental group (62 ± 10) %was lower than that in the control group (89 ± 12) %(P< 0.05), whereas up-regulation of miR-181a with miR-181a agomir could improve the migration of H929 cells, and the proportion of migrated cells in the experimental group (242 ± 9) % was higher than that in the control group (98 ± 8)%(P<0.01). Conclusions The high expression of miR-181a expressed highly by MM cells may promote the proliferation, migration and drug resistance of myeloma cells, indicating that miR-181a could be an important prognostic biomarker candidate, and the application of gene silencing may improve the prognosis of MM.

Objectives:To analyze the correction of postoperative coronal imbalance in adult degenerative scoliosis patients undergoing spinal fusion surgery,and to explore the impact of leveling of the most tilted vertebra(L4 or L5)in the lumbosacral curve on the correction of coronal imbalance.Methods:A retrospective analysis was conducted on 61 patients(17 male and 44 female;aged 51 to 78 years,mean 63.57±8.41 years;body mass index 15.79-35.55kg/m2,mean 25.83±3.69kg/m2)who underwent degenerative scoliosis correction surgery between January 2019 and March 2023 in Tianjin Union Medical Center.The patients were catego-rized based on the Obeid classification and the length of fusion segments into concave long segment group(17 cases),concave short segment group(15 cases),convex long segment group(15 cases),and convex short seg-ment group(14 cases).Coronal radiographic parameters[Cobb angle,coronal balance distance(CBD),coronal tilt angle]and sagittal radiographic parameters(sagittal vertical axis,thoracic kyphosis,lumbar lordosis,pelvic inci-dence,pelvic tilt,sacral slope)were measured on full spine anteroposterior and lateral X-rays preoperatively and on 3d postoperatively.The postoperative coronal and sagittal imbalance corrections were compared be-tween groups.Pearson correlation analysis was performed to investigate the relationship between the correction ratio of the maximal coronal tilt angle in the lumbosacral curve(L4 or L5)and postoperative CBD.The opera-tive time,blood loss,and visual analogue scale(VAS)pre-and postoperatively were compared between the four groups.Results:The postoperative coronal Cobb angle,CBD,and maximal coronal tilt angle at L4 or L5 of the concave long segment group,convex long segment group,and convex short segment group were statisti-cally different from those before operation(P＜0.05).In the concave short segment group,the postoperative coronal Cobb angle and maximal coronal tilt angle at L4 or L5 were significantly different from those before operation(P＜0.05),while no significant difference was found between pre-and postoperative CBD(P=0.085).Pearson correlation analysis revealed a significant negative correlation(r=-0.570,P＜0.01)between postoperative CBD and coronal tilt correction ratio in the convex group,with the linear regression equation being postoper-ative CBD=32.81-28.38 x coronal tilt correction ratio.When the correction ratio of the maximal coronal tilt angle at L4 or L5 exceeding 45％in convex coronal malalignment patients,it's predicted that the CBD could be corrected to within 20mm.No significant correlation was found between postoperative CBD and the coronal tilt correction ratio in the concave group(r=0.098,P=0.59).Patients in both the convex and concave short segment groups had shorter operative time and less blood loss than long segment groups.Postoperative VAS scores improved significantly in all groups(P＜0.05).Conclusions:In adult degenerative scoliosis patients with convex coronal malalignment,both short and long segment fusions are capable of correcting coronal imbalance.Concave coronal malalignment patients require long segment fusion for the correction of coronal imbalance.To achieve optimal coronal balance in convex coronal malalignment patients,a correction ratio of the maximal coronal tilt angle at L4 or L5 shall exceed 45％.

Objective:To analyze the relationship between perineural invasion and other clinicopathological factors and its effect on the prognosis of gastric cancer.Methods:The clinicopathological data of 665 patients with gastric cancer were retrospectively analyzed. According to the presence of perineural invasion, the patients were divided into perineural invasion positive group and perineural invasion negative group. The relationship between perineural invasion and other clinicopathological factors and its effect on the prognosis of gastric cancer were analyzed. After eliminating the potential confusion bias between the two groups by propensity score matching (PSM) , the differences of 5-year cumulative survival rate between the two groups of gastric cancer patients were compared.Results:The incidence of perineural invasion was 17.0% (113 cases) . The binary logistic regression analysis showed that the depth of tumor invasion and vascular tumor thrombus were independent factors influencing the occurrence of gastric cancer perineural invasion (all P<0.001) . Univariate analysis showed that age (>60 years) , tumor diameter (>4 cm) , borrmann classification, depth of invasion, lymph node metastasis, TNM stage, degree of differentiation, vascular tumor thrombus, perineural invasion, tumor nodule, tumor site, resection site, and surgical operation were the influencing factors for the prognosis of patients with gastric cancer ( P<0.05) , but multivariate analysis showed that age (>60 years) , tumor diameter (>4cm) , depth of invasion, lymph node metastasis, and positive vascular tumor thrombi were independent risk factors affecting the prognosis of gastric cancer patients ( P<0.05) .However, perineural invasion cannot be an independent factor influencing the poor prognosis of gastric cancer in a multivariate analysis. Survival analysis was performed after propensity matching scores, and it was found that there was no statistically significant difference in the five-year survival rate between the perineural invasion positive group and the perineural invasion negative group (34.6% vs 43.0%; χ2=1.713; P=0.191) ,and there was no significant difference in the survival curve analysis between the two. Conclusion:Most patients with gastric cancer of perineural invasion have poor prognosis, but perineural invasion cannot be an independent prognostic factor for the prognosis of gastric cancer.

Objective: To identify the expression pattern of TIM-3 in natural killer/T-cell lymphoma (NK/TCL) cell lines, and to investigate the effect and mechanism of its ligand galectin-9 (GAL-9) inducing apoptosis of NK/TCL cell lines. Methods: Expression of TIM-3 in NK cell of peripheral blood from healthy donors and NK/TCL cell lines (SNK-1、SNK-6、SNT-8) was detected by Western blotting. After being treated with rhGAL-9 at various concentrations for 24h, the cell proliferation ability was analyzed with CCK-8 assay. Apoptosis ratio of the cells was determined by flow cytometry. Expressions of caspase-3, PARP and their cleavages were detected by Western blotting; moreover, phosphorylation levels of proteins in MAPK signaling pathway were also detected by Western blotting. Results: The expression of TIM-3 in SNK-1, SNK-6 and SNT-8 cell lines was significantly higher than that of NK cells from healthy donors (P<0.05). CCK-8 result showed that rhGAL-9 obviously inhibited the proliferation of NK/TCL cell lines in a concentration dependent manner. Flow cytometry showed that rhGAL-9 induced the apoptosis of NK/TCLcells; and Western blotting proved that the expression of cleaved caspase-3, cleaved-PARP, and p-JNK in MAPK signaling pathway were significantly elevated. Conclusion: TIM-3 was over-expressed in NK/TCL cell lines, and its ligand galectin-9 induced cell apoptosis probably through the activation of JNK kinase pathways.