CMTM家族作为一个新的基因家族，在免疫、生殖等系统以及多种肿瘤的发病机制中起到重要作用。CMTM家族中， CMTM1可影响细胞增殖，导致肿瘤发生；与非小细胞肺癌（NSCLC）患者的化疗耐药和预后有关。CMTM2通过AP-1、CREB通 路一定程度上影响HIV-1转录，同时在男性生殖系统中起重要作用。CMTM3等位基因失活或甲基化使其丧失对细胞增殖的负 性调控能力，是胃癌独立的预后指标。CMTM4调控细胞周期影响肿瘤细胞增殖，通过对PD-L1的协同保护参与免疫逃逸。 CMTM5在许多肿瘤中沉默表达，参与肿瘤发生发展相关的信号通路。CMTM6协同PD-L1参与免疫逃逸，是潜在的免疫治疗靶 点。CMTM7在NSCLC中通过Rab5控制EGFR-AKT信号影响肿瘤发展，且与胃癌发展相关。CMTM8通过MARVEL区域影响 EGFR和相关信号通路，调控细胞增殖、分化和凋亡等。上述重要发现，为研究肿瘤的发生发展及肿瘤基因治疗提供了新思路。

Patients with knee osteoarthritis increase gradually.Arthroscopic debridement has achieved good results in clinical treatment at home and abroad in recent years.The technology is not only easy to operate at a low cost, it but also can directly improve the internal environment and function of the knee joints, cut off the vicious circle of joint cavity, which can provide a good environment for the normal production of joint fluid after a large amount of saline lavaging knee joint cavity during operation. This review summarizes the clinical curative effect of knee osteoarthritis with arthroscopic debridement in recent five years and provides the guidance and reference for the researchers.

Objective To investigate the relevance between the times of cytokine-induced killer cell (CIK cell) adoptive immunotherapy and the survival of the elderly patients with gastric cancer.Methods Lymphocyte separation medium was used to isolate mononuclear cells, and then the cultured CIK cells were infused back into the patients with gastric cancer. A retrospective cohort study was adopted by using Kaplan-Meier to estimate median survival time and survival rate, using Log-rank test to analyze the impact of clinical factors on survival rate, and using RR and 95% CI to estimate the contact intensity of death outcome, survival time and CIK cell treatment. Results There were nostatistically significant differences in gender,age, tumor site, histological type, invasion depth, lymph node metastasis, pathological grade, tumor size or tumor distribution between chemotherapy group and CIK treatment group (all P＞0.05). The median survival time of patients with gastric cancer was significantly longer in CIK treatment group than in chemotherapy group (61 vs. 21, χ2=10.215, P=0.001). Compared with the patients treated by chemotherapy alone, the increased times of CIK treatment induced more survival rate and 2-5 years life spans (χ2=12. 461, P=0.006). Conclusions With the treatment that CIK cells are infused back into the elderly patients with gastric cancer, the risk of death is reduced, and the lifespan is prolonged, which is associated with the CIK cell treatment times.

Objective To investigate the effects of down-regulation of lemur tyrosine kinase 3 (LMTK3) on the cytobiological behaviors of human lung cancer A549 cells.Methods The expression of LMTK3 in A549 cells was interfered with small hair RNA (shR-NA),and the expression level of LMTK3 was determined by RT-PCR.The effects of LMTK3 on the proliferation,migration,cell cycle and apoptosis of A549 cells were determined by the CCK-8 assay,scratch assay,Transwell assay and flow cytometry,respectively.Results The shLMTK3 cells with stably low expression of LMTK3 and control cells (Scramble) with normal expression of LMTK3 were successfully obtained.The relative proliferation rates of shLMTK3 cells cultured for 24,48 and 72 hours were significantly lower than those in the control cells (0.305 ±0.018 vs 0.354 ±0.011,t =5.24,P＜0.01;0.461 ±0.044 vs 0.551 ±0.027,t =3.91,P ＜0.01;0.74 ± 0.029 vs 0.881 ± 0.028,t =7.70,P ＜ 0.01).The relative migration rate of shLMTK3 cells 24 hours after scratching was significantly lower than that of control cells (0.51 ±0.096 vs 1.00 ± 0.029,t =4.81,P ＜ 0.01).Transwell assay showed that the number of migration cells in shLMTK3 group was significantly less than that in Scramble group (161 ±9.29 vs 308.66 ± 17.60,t =7.42,P ＜ 0.05).The results of flow cytometry showed that shLMTK3 cells were blocked at G1 phase (t =4.35,P ＜ 0.05),and that the inhibition of LMTK3 had no influence on the apoptosis of A549 cells.Conclusion Down-regulation of LMTK3 expression in human lung cancer A549 cells may inhibit the proliferation and migration of A549 cells significantly,indicating that the abnormal expression of LMTK3 in lung carcinoma cells may regulate the biological behaviors and progression of tumors.

Objective To study the in vitro anti-tumor activity of dendritic cells (DCs) loading with antigen produced by radiofrequency ablation of tumor lysate in situ combined with cytokine-induced killer cells (CIK).Methods CIK ceils derived from BALB/C mouse spleen and DCs derived from bone marrow were prepared,and experimental model of murine colon carcinoma were established for radiofrequency ablation.The supernatant of tumor tissue in situ lysis after repeated freezing and thawing were tested by lowry protein quantitative statutory,amounting to a final concentration of 5 μg/ml,then load to the first 5 days of culture DCs (Ag-DC),2 days later,co-cultured with CIK cells after the first seven days of culture 48 h (Ag-DC-CIK).Flow cytometry was used to analyze costimulatory molecules on the surface of the cells,and CCK-8 assay to detect in vitro cytotoxic activity.Results The DCs loading with antigen resulted in an increase in the proportion of CD86 + CD11 c +,MHC Ⅱ + CD11 c + and MHC Ⅱ + CD80 + cells.The main effector cells of CIK cells were CD3 + NK1.1 + cells.The percentage of CD3 + NK1.1 + cells was 1.45％ on the first day of the culture ; while when they had been cultured for 7 days,the percentage CD3 + NK1.1 + significantly increased to 36.9％.The cytotoxicity of Ag-DC-CIK cells toward C26 cells was much more efficient than that of DC-CIK,CIK cells.The cytotoxic activity of the former was significantly lower than the latter and the same target ratio.When the ratios of effector cells to target cells were 5 ∶ 1,the cytotoxic activity of Ag-DC-CIK cells against C26 cells was (74.9 ± 3.5) ％,; while the DC-CIK was (71.2 ± 2.1) ％ and the CIK cells was (68.7 ± 2.9) ％.The difference was statistically significant(F =7.007,P =0.007).When the ratios of effector cells to target cells were 10 ∶ 1,the cytotoxic activity of Ag-DC-CIK cells against C26 cells was (82.3 ± 4.5) ％,while the DC-CIK cells was (77.1 ± 5.1) ％,and the CIK cells was (72.7 ± 2.8) ％.The difference was statistically significant (F =7.727,P =0.005).When the ratios of effector cells to target cells were 20 ∶ 1,the cytotoxic activity of Ag-DC-CIK cells against C26 cells was (83.2 ± 1.9) ％,while the DC-CIK cells was (77.2 ± 4.2) ％,and the CIK cells was (73.0 ± 2.6) ％.The difference was statistically significant (F =16.594,P =0.000).Conclusion DCs loading with antigen produced by radiofrequency ablation of tumor in situ pyrolysis products can improve in vitro cytotoxic activity combined with CIK cells,which can provide a new comprehensive cancer treatment strategy.

Objective To evaluate the clinical efficacy and adverse effects of brucea javanica oil emulsion combined with DP chemotherapy in treating advanced non-small-cell lung cancer.Methods Totally 48 patients with advanced non-small-cell lung cancer were divided into two groups randomly by mechanical sampling method.Twenty-four cases in treatment group were treated by brucea javanica oil emulsion combined with DP chemotherapy, while 24 cases in control group were treated by DP chemotherapy only.The clinical effects were evaluated after treatment of two cycles.Results The short-term effective rate was 54.2% (13/24) in treatment group and 45.8% (11/24) in control group, and there was no significant difference between two groups ( χ2 = 0.333, P = 0.564).The rate of increased and stable life quality was 87.5%(21/24) in treatment group and 58.3%(14/24) in control group,and there was significant difference between two groups (χ2 = 5.169,P = 0.023).The rate of increased and stable weight was 79.2% (19/24) in treatment group and 45.8%( 11/24) in control group, and there was significant difference between two groups (χ2 = 5.689,P = 0.017).The incidence of nausea or vomiting was 45.8% (11/24) in treatment group and 41.7%( 10/24 ) in control group, and there was no significant difference between two groups (χ2 = 0.085, P = 0.771 ).Compared with those in control group, patients in treatment group had less adverse effects in decreasing of peripheral blood leucocytes and showed better immune function.Conclusion Brucea javanica oil emulsion combined with DP chemotherapy in treating advanced non-small-cell lung cancer has good clinical effect, especially enhances the quality of life, improves immune function and decreases the adverse effects of chemotherapy.

Objective To evaluate the feasibility of the novel staining protocol based on protein L in flow cytometry to detect the expression of chimeric antigen receptor (CAR) on the surface of T cells from patients.Methods The peripheral blood mononuclear cells(PBMCs) were collected from 2 patients with CD19 + lymphoma by hemapheresis and T cells were purified by magnetic bead selection.CAR was transfected with retroviruses targeting CD19 and CD22 into T cells to induce chimeric antigen receptor-modified T cells (CAR-T cells).The single-chain fragments of antibody molecules on the surface of CAR-T cells were labeled by biotinylated protein L-streptavidin-PE system.The proportions of activated CD19-CAR and CD22-CAR positive T cells in the culture were detected by flow cytometry.The results were compared with those detected by conventional flow cytometry method based on Anti-IgG staining.Results The expression of CAR on the surface of CAR-T cells was successfully detected by flow cytometry protocol based on the staining of single-chain variable fragment (scFv)-biotinylated protein L-streptavidin-PE.The percentages of CD19-CAR-T cells from the 2 patients were 71.6％ vs 64.2％ and 49.3％ vs 43.8％ in the protein L group and the Anti-IgG control group respectively,and the percentages of CD22-CAR-T cells were 53.1％ vs 46.3％ and 56.5％ vs 64.0％.The results of the both groups were similar.Conclusion The staining protocol based on protein L could be used as a routine staining method in flow cytometry for the detection of CAR-T cells.

Objective To purify the anti-T cell immunoglobulin mucin ( TIM)-3 monoclonal antibody 4E8 and examine its biological function in vitro. Methods The mouse monoclonal antibody against human TIM-3, clone 4E8, was obtained by standard protocol for monoclonal antibody purification. The cell lines expressing human TIM-3 molecule were obtained by cell transfection technique. We ex-amined the ability of 4E8 binding to human TIM-3 by flow cytometry. The ability of 4E8 blocking the binding of fusion protein TIM-3 Ig-huFc with phosphatidylserine( PtdSer) , the apoptotic cell surface TIM-3 ligand, was also analyzed by flow cytometry. Mixed lympho-cyte reaction ( MLR) and ELISA assays were used to determine the effect of TIM-3 monoclonal antibody ( 4E8) on IFN-γsecretion in CD4+ T cells. Results 4E8 specifically bound to human TIM-3 but could not block the binding of TIM-3 to Ptdser. Compared with the negative control (IFN-γ secretion: 958.3±153.2), 4E8 enhanced the ability of CD4+ T cells to secrete IFN-γ in MLR (4E8 of 10μg/mL group:IFN-γ secretion 2563±150.3 and 4E8 of 3.33 μg/mL group:IFN-γ secretion 1981±211.5) with statistically signifi-cant difference ( P<0.05) . In addition, the combined application of 4E8 with the anti-programmed death-1 ( PD-1) monoclonal anti-body nivolumab showed synergistic effects for increasing IFN-γ secretion in MLR assay ( 4E8 of 10 μg/mL group: IFN-γ secretion 3049±80.5 and 4E8 of 0.33μg/mL group:IFN-γsecretion 1957±321.3) as compared with 4E8 alone (10μg /mL group:IFN-γse-cretion 2563±150.3 and 0.33 μg/mL group:IFN-γ secretion 844±76.2) with statistically significant difference (P<0.05). Conclu-sion We successfully obtained a 4E8 clone of monoclonal antibody to human TIM-3 which may enhance the capacity of IFN-γsecre-tion from CD4+ T cells. The effect of enhancing IFN-γ secretion of CD4+T cells by TIM-3 monoclonal antibody was independent from blocking the binding of TIM-3 with Ptdser.

Objective To construct a prognostic nomogram for predicting the prognosis of patients with colorectal cancer ( CRC) , and verify its accuracy. Methods The clinical pathologic data from 438 CRC patients hospitalized in the Third Affiliated Hospital of Soo-chow University during January 2006 and May 2013 were retrospectively analyzed. The independent risk factors for predicting the prog-nosis of CRC were determined by the univariate and multivariate regression model. The prognostic nomogram was established by the R-language software. Then, the nomograms of postoperative 3-year and 5-year disease free survivals ( DFS) were drawn, and compared with the actual status. The internal validation and accuracy of the nomogram were determined by the Bootstrap method and the calculat-ed concordance index ( C-index) , respectively. The sensitivity and specificity of the nomogram for predicting the 3-year and 5-year DFS were compared with those of TNM system established by the American Joint Committee On Cancer (AJCC) (7th ed.) by using the time-dependent ROC curve. Results Among 438 CRC patients, the metastasis of CRC occurred in 233 patients, including 105 liver metas-tasis and 57 lung metastasis. Multivariate COX regression analysis showed that tumor differentiation degree, TNM stage, serum CEA level, serum CA19-9 level, neutrophil lymphocyte ratio ( NLR) and P53 level were the independent risk factors of CRC. The C-index of the constructed nomogram for predicting the survival rate of CRC patients was 0.678. The predicted 3-year and 5-year DFS by the no-mogram were highly coincident with the actual status. The analysis results of the time-dependent ROC curve showed that the sensitivity and specificity of the established nomogram for predicting the postoperative 3-year and 5-year DFS were higher than those of AJCC-TNM stage.Conclusion The established nomogram may accurately predict the prognosis of CRC patients, which may be helpful for clinicians to follow up or make beneficial treatment for CRC patients.

Objective To explore a new efficient extraction method of dorsal root ganglions(DRGs)of rats by exploring and extracting DRGs reversely along the nerve traveling pathway. Methods The DRGs were extracted by the traditional method of opening the intervertebral foramina and the new method, extracting DRGs reversely along the nerve traveling pathway,respectively. The time consuming and the number of intact DRGs obtained with these two method were compared. Results The number of intact DRGs(L3-5segments,both sides)extracted from each rat with the traditional method was(3.08 ± 1.31),and the average time consuming of each DRG was(5.58 ± 1.21)min. As for the new method ,the number of intact DRGs extracted from each rat was(4.29 ± 1.08), and the average time consuming was(1.69 ± 0.91)min,significantly better than that of the traditional method(P < 0.05 for both). Conclusions The new method of exploring and extracting DRGs reversely along the nerve traveling pathway is more efficient for obtaining intact DRGs of rats,providing more useful tissue materials for subsequent culture and morphological studies of DRG cells.