OBJECTIVES: To investigate the subcellular localization and biological functions of transmembrane protein 240 (TMEM240). METHODS: NCBI BLAST and TMHMM bioinformatics software were used for protein sequence analysis and prediction of transmembrane domain of TMEM240. Brain tissues from male C57BL/6 mice (18-20 days old) were examined for distribution of TMEM240 using in situ hybridization, and qPCR and Western blotting were used to detect TMEM240 expression in different mouse tissues and in cortical neurons at different time points (n=3). In the in vitro experiment, HepG2 and Neuro-2a cells were transfected with plasmids for overexpression of TMEM240, and subcellular localization of TMEM240 was analyzed using cell imaging. In primary cultures of cortical neurons isolated from C57BL/6 mice, TMEM240 expression and its biological functions were investigated using qPCR, Western blotting, and immunofluorescence staining. RESULTS: Human and mouse TMEM240 proteins share a 97.69% similarity in the protein sequences, and both are transmembrane proteins with two transmembrane domains. TMEM240 mRNA and protein were highly expressed in mouse brain tissues and cortical neurons. In isolated mouse cortical neurons, TMEM240 expression reached the peak level after primary culture for 9 days and distributed in scattered spots within the cells. In HepG2 cells, TMEM240 was characterized as intracellular membrane structures and showed 80% colocalization with peroxisomes. In Neuro-2a cells, TMEM240 overexpression caused significant enhancement of the expressions of Rho GDP dissociation inhibitor β (ARHGDIB) at both the mRNA and protein levels. CONCLUSIONS TMEM240 is a novel intracellular subcellular structure specifically expressed in neurons with significant potential for targeted cellular function regulation.
Animals ; Humans ; Mice ; Peroxisomes/metabolism* ; Membrane Proteins/genetics* ; Mice, Inbred C57BL ; Neurons/metabolism* ; Male ; rho-Specific Guanine Nucleotide Dissociation Inhibitors ; Hep G2 Cells ; Brain/metabolism*

Objective To analyze the correlation between lipid metabolism indexes,serum gamma-glutamyl transpeptidyase(γ-GGT)and coronary heart disease(CHD)complicated with coronary artery calcification(CAC).Methods A total of 300 CHD patients admitted in this study were divided into the CAC group(n=193)and the non-CAC group(n=107).Clinical data of the two groups were compared,including high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),total cholesterol(TC),triglyceride(TG),Apolipoprotein A1(Apo-A1),Apo-B(APO-B)and γ-GGT.The influencing factors of CAC were analyzed by multiple Logistic factors.And a nomogram prediction model was established.Results The basic data of the two groups were compared.Patients of the CAC group was older,had higher proportion of patients with hypertension and diabetes,had higher levels of LDL-C,TC,Apo-B and γ-GGT and lower level of Apo-A1 than those of the non-CAC group(P＜0.05).The results of Logistic multivariate regression analysis showed that advanced age,combined history of diabetes,elevated LDL-C,TC,Apo-B and γ-GGT were risk factors of CHD complicated with CAC,while elevated Apo-A1 was the protective factor of CHD complicated with CAC(P＜0.05).The AUC of the constructed nomogram model was 0.880(95%CI:0.840-0.919),which showed good distinguishing ability.Conclusion CHD complicated with CAC is related to lipid metabolism and γ-GGT level.The nomogram model constructed based on influencing factors can be used for clinical early warning of CAC risk.

Objective To analyze the correlation between lipid metabolism indexes,serum gamma-glutamyl transpeptidyase(γ-GGT)and coronary heart disease(CHD)complicated with coronary artery calcification(CAC).Methods A total of 300 CHD patients admitted in this study were divided into the CAC group(n=193)and the non-CAC group(n=107).Clinical data of the two groups were compared,including high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),total cholesterol(TC),triglyceride(TG),Apolipoprotein A1(Apo-A1),Apo-B(APO-B)and γ-GGT.The influencing factors of CAC were analyzed by multiple Logistic factors.And a nomogram prediction model was established.Results The basic data of the two groups were compared.Patients of the CAC group was older,had higher proportion of patients with hypertension and diabetes,had higher levels of LDL-C,TC,Apo-B and γ-GGT and lower level of Apo-A1 than those of the non-CAC group(P＜0.05).The results of Logistic multivariate regression analysis showed that advanced age,combined history of diabetes,elevated LDL-C,TC,Apo-B and γ-GGT were risk factors of CHD complicated with CAC,while elevated Apo-A1 was the protective factor of CHD complicated with CAC(P＜0.05).The AUC of the constructed nomogram model was 0.880(95%CI:0.840-0.919),which showed good distinguishing ability.Conclusion CHD complicated with CAC is related to lipid metabolism and γ-GGT level.The nomogram model constructed based on influencing factors can be used for clinical early warning of CAC risk.

Objective]To explore the metastasis regulatory of lymph node of papillary thyroid cancer and to analyze the influence factors.[Methods]Clinical data of 375 papillary thyroid cancer patients at our hospital between Jun 2011 and Sep 2015 were retrospectively reviewed and summarized the metastasis regulatory of lymph nodes and the tumor characteristics.[Results]All selected patients were diagnosed papillary thyroid cancer. The Total metastasis rate of cervical lymph node was 67.47%,the metastasis rate of region Ⅵ lymph nodes was 64.27%;the metastasis rate of region Ⅱ~Ⅴ lymph nodes was 36.53%. The metastasis rate of lymph nodes of the patients with tumor diameter over 1 cm,breaking through thyroid membrane and invading the cervical muscle were significantly increased(P < 0.05).[Conclusion]The central group lymph nodes were the most metastasis region of papillary thyroid cancer and should routinely be dissected by the first time of surgery. When the tumor diameter greater than 1 cm or cancer breakthrough thyroid membrane and/or invading the cervical muscles ,the ipsilateral lateral neck lymph nodes should be dissected at the same time.

The high morbidity and mortality of non-small cell lung cancer (NSCLC) did influence the quality of life of tumor patients world-wide. There is an urgent need to develop new therapies that have high anti-tumor activity and low toxicity side effects. It is widely accepted that autophagy can play diverse roles in carcinogenesis, such as induces pro-death of lung cancer cells or helps the escape from cell death, making it become a proper anticancer target. It's believed that various monomers of Chinese traditional medicine closely correlates to anti-NSCLC activities, and that even could affect the acquired multiple drug resistance (MDR). Furthermore, autophagy might be the underling mechanisms which could play a role as the candidate targets of natural active compounds. Recent studies of terpenoids, alkaloid, dietary polyphenols, saponins and other active ingredients that extracted from a large variety of herbs suggest that different monomer compounds could either regulate the activity of pro-death autophagy or influence the level of protective autophagy of NSCLC cells, thus changing their drug sensitivity and cell viability. This paper aims to give a systemic description of the latest advances about natural compounds and their derivatives that involved in tumorigenesis of NSCLC via inducing the autophagy.

ObjectiveTo evaluate the therapeutic effects of radiofrequency ablation and surgical resection for liver metastasis from breast cancer.MethodsClinical data of 15 patients with liver metastasis from breast cancer admitted and treated in the Third Affiliated Hospital of Sun Yat-sen University between January 2010 and May 2014 were retrospectively studied. All patients were females with the age ranging from 33 to 66 years old and the median of 49 years old. The informed consents of all patients were obtained and the local ethical committee approval had been received. According to the different therapeutic regimen, the patients were divided into radiofrequency ablation + chemotherapy group (ablation group,n=9) and surgery +chemotherapy group (surgery group,n=6). The conditions of two groups during the perioperative period were observed and the survival analysis was performed. The observations during perioperative period between two groups was compared usingt test, the rates was compared using Fisher's exact test and the survival analysis was conducted using Kaplan-Meier method and Log-rank test.ResultsThe mean operation time of the ablation group was (26±5) min, which was significantly shorter than (151±27) min of the surgery group (t=-18.69,P<0.05). No case in the ablation group received blood transfusion while 3 cases in the surgery group received blood transfusion during the operation. The tumor clearance rate of the surgery group was 100%,which was the same with that of the ablation group. The postoperative hospital stay of the ablation group was (6±2) d, which was signiifcantly shorter than (11±5) d of the surgery group (t=-3.70,P<0.05). The incidence of postoperative complications of the ablation group was 4/9, which was signiifcantly lower than 5/6 of the surgery group (P=0.02). The median survival time of the ablation group and the surgery group was respectively 33, 23 months. The 1-, 2- and 5-year cumulative survival rate were respectively 88.9%, 66.7% and 22.2% for the ablation group and were respectively 82.3%, 50.0% and 0 for the surgery group and no significant difference was observed between two groups (χ2=1.53,P>0.05).ConclusionsThe radiofrequency ablation for patients with liver metastasis from breast cancer can have the same therapeutic effect with surgical resection, and has advantages of short operation time, low incidence of postoperative complications and short postoperative hospital stay.

Objective To establish a NOD/SCID mouse model with human immune reconstitution and observe its immune response to human triple-negative breast cancer xenograft. Methods Twenty-four NOD/SCID mice without immune leakage were subjected to cyclophosphamide (CTX) treatment 3 days prior to immune reconstitution with human peripheral blood mononuclear cell (PBMC) injection and subcutaneous transplantation of human triple-negative breast cancer MDA-MB-231 cells, CTX treatment and PBMC injection without tumor cell transplantation, MDA-MB-231 cell transplantation only, or no treatments. The tumor growth and immune responses of the mice were observed at regular intervals. Results Compared with the tumor-bearing mice, the tumor-bearing mice with immune reconstitution showed prolonged incubation period of tumor formation, slower tumor growth rate and increased survival rate. Human IgG and CD3+T cells were detected in the peripheral blood of the mice 1 week after human PBMC injection. The percentage of CD3+T cells in the spleen cells was 55.3%at 9 weeks in tumor-bearing mice with immune reconstitution and 52.7% in tumor-bearing mice without immune reconstitution. The spleen index of the tumor-bearing mice with immune reconstitution was much higher than that in mice with only immune reconstitution and the control mice (9.64 vs 3.82±0.31 and 1.51±0.14 mg/g). Conclusion A stable NOD/SCID mouse model with immune reconstitution has been established successfully, which shows immune responses to triple-negative breast cancer xenografts and allows studies of immunological therapy study of triple-negative breast cancer.

Objective To establish a NOD/SCID mouse model with human immune reconstitution and observe its immune response to human triple-negative breast cancer xenograft. Methods Twenty-four NOD/SCID mice without immune leakage were subjected to cyclophosphamide (CTX) treatment 3 days prior to immune reconstitution with human peripheral blood mononuclear cell (PBMC) injection and subcutaneous transplantation of human triple-negative breast cancer MDA-MB-231 cells, CTX treatment and PBMC injection without tumor cell transplantation, MDA-MB-231 cell transplantation only, or no treatments. The tumor growth and immune responses of the mice were observed at regular intervals. Results Compared with the tumor-bearing mice, the tumor-bearing mice with immune reconstitution showed prolonged incubation period of tumor formation, slower tumor growth rate and increased survival rate. Human IgG and CD3+T cells were detected in the peripheral blood of the mice 1 week after human PBMC injection. The percentage of CD3+T cells in the spleen cells was 55.3%at 9 weeks in tumor-bearing mice with immune reconstitution and 52.7% in tumor-bearing mice without immune reconstitution. The spleen index of the tumor-bearing mice with immune reconstitution was much higher than that in mice with only immune reconstitution and the control mice (9.64 vs 3.82±0.31 and 1.51±0.14 mg/g). Conclusion A stable NOD/SCID mouse model with immune reconstitution has been established successfully, which shows immune responses to triple-negative breast cancer xenografts and allows studies of immunological therapy study of triple-negative breast cancer.

Objective To investigate the role of SIAH2 protein in the occurrence and development of hepatocellular carcinoma (HCC). Methods Human HCC Bel-7404 cells in logarithmic growth phase were inoculated. Empty carrier small interference ribonucleic acid (siRNA) and SIAH2 siRNA were transfected in human HCC Bel-7404 cells. Then the cells were divided into 2 groups:control-siRNA group and SIAH2-siRNA group. Theβ-actin was taken as control, and the expression of SIAH2 protein in human HCC Bel-7404 cells of 2 groups was detected by Western Blot. The proliferation of cells in 2 groups was detected by methyl thiazolyl tetrazolium (MTT) assay. Cell migration in 2 groups was observed by cell scratch test. Cell invasion in 2 groups was observed by Transwell assay. The data in 2 groups were compared using t test. Results The average relative expression of SIAN2 in control-siRNA and SIAH2-siRNA group were 0.71±0.02, 0.33±0.01 respectively. The expression of SIAN2 in SIAH2-siRNA group decreased obviously compared with that in control-siRNA group (t=-4.629, P<0.05). The proliferation rate in SIAH2-siRNA group also decreased obviously. The cell migration rate in SIAH2-siRNA group[(14.3±0.4)%] was significantly lower than that in control-siRNA group [(45.3±0.4)%]( t=-3.689, P<0.05). The membrane permeating cell count in SIAH2-siRNA group (122±7) was signiifcantly less than that in control-siRNA group (563±10) (t=-3.428, P<0.05). Conclusion SIAH2 protein can promote the proliferation, migration and invasion of HCC cells and thus accelerate the occurrence and development of HCC.

Objective To investigate the curative effect of radiofrequency ablation (RFA) combined with chemotherapy for liver metastases from breast cancer. Methods Clinical data of 6 patients with liver metastases from breast cancer in the Third Affiliated Hospital of Sun Yet-sen University from January 2009 to December 2012 were analyzed retrospectively. The informed consents of all patients were obtained and the ethical committee approval was received. All the patients were female with the age ranging from 35 to 65 years old and a median of 54 years old. The liver metastases were all single metastases. The tumor diameter was 1.5 to 4.5 cm with a median of 2.5 cm. The patients received modified radical mastectomy 6 to 30 months before treatments and all received chemotherapy after the operations. The liver metastases were confirmed by ultrasound, CT and MRI. RFA was performed percutaneously under the guidance of ultrasound on the patients. After 1 week of RFA treatment, individual chemotherapy was given to the patients according to their previous chemotherapy regimen and the situation of liver metastases. The focus necrosis was checked by contrast enhanced CT or MRI examination 1 month after the RFA treatment. The curative effect of RFA was observed based on the imaging examinations. According to the patients' survival and tumor recurrence, survival analysis was conducted by drawing Kaplan-Meier curves. Results After the first RFA on 6 cases, the liver metastases in 5 cases were observed with total necrosis, and 1 case with partial necrosis. The one with partial necrosis then received the second RFA. Local recurrence of the tumor was observed in 1 case during the follow-up. All the patients survived with a median tumor-free survival time of 18 months. Conclusions The combination theraphy of RFA and chemotherapy has a definite effect for liver metastases from breast cancer and is a safe, effective comprehensive therapeutic regimen.