Undifferentiated embryonic stem (ES) cells can be maintained in vitro if cultured in the presence of the cytokine leukaemia inhibitory factor (LIF). ES cells can also differentiate in vitro. A particularly efficient method for inducing ES cell differentiation is to culture ES cells as aggregates in the absence of LIF. Under these conditions they form structures known as embryoid bodies (EBs). However the current protocols for EB formation are still diverse. In order to facilitate further study, we carefully controlled the culture conditions for EB formation, and here we report an efficient protocol by which uniformly differentiated EBs were obtained, monitored by measuring the differentiation of beating cardiomyocytes. Furthermore, by using this protocol we observed in long-term cultured plating EBs (> 60 days) there still exist cell colony with pluripotency. This observation raised a potential possibility that ES cells may keep pluripotent in a niche provided by differentiated cells.
Animals ; Cell Differentiation ; physiology ; Cells, Cultured ; Culture Media ; Embryonic Development ; physiology ; Embryonic Stem Cells ; cytology ; Leukemia Inhibitory Factor ; pharmacology ; Mice ; Stem Cell Niche ; physiology ; Time Factors

Objective: To investigate the clinical value of the serum new molecular markers, soluble triggering receptor expressed on myeloid cells-1(sTREM-1)and soluble hemoglobin scavenger receptor(sCD163), in the diagnosis of sepsis in elderly patients with burns. Methods: A total of 58 inpatients with burns from Jun 2017 to June 2018 were enrolled in the study.Patients were divided into three groups: the sepsis group(n=12), the localized infection group(n=21)and the non-infection group(n=29). The levels of sTREM-1 and sCD163 were determined by enzyme-linked immunosorbent assays(ELISAs). The clinical diagnostic value of sTREM-1 and sCD163 was assessed by receiver operating characteristic(ROC)curve analysis. Results: There was a statistically significant difference in the levels of sTREM-1 and sCD163 at day 1 between the three groups(F=20.994 and 38.363, P<0.01). Serum levels of sTREM-1 and sCD163 were higher in the sepsis group than in the localized infection group and the non-infection group.Serum levels of sTREM-1 and sCD163 were higher in the localized infection group than in the non-infection group.Serum levels of sTREM-1 and sCD163 were lower at day 7 than those at day 1 in all groups(F=21.242 and 41.035, P<0.01). Serum sTREM-1 levels were positively correlated with serum sCD163 levels(r=0.609, P=0.000). The AUC of sTREM-1 and sCD163 for the diagnosis of sepsis was 0.880(95%CI: 0.816~0.926). Conclusions Serum levels of sTREM-1 and sCD163 are elevated with increasing degrees of infection.Monitoring serum sTREM-1 and sCD163 levels is helpful for the diagnosis of sepsis in elderly patients with burns.

Objective To explore the potential impact pathways of self-management behavior based on COM-B model,in maintenance hemodialysis(MHD)patients,and provide reference for the intervention of self-manage-ment behavior.Methods Judgment sampling was used to select 350 MHD patients undergoing dialysis at the hospital in a certain district of Guangzhou,and a questionnaire survey was conducted among them.Smart PLS software was used to construct a partial least squares structural equation model and perform path analysis.Results Self-manage-ment behavior score of MHD patients was(57.06±13.28).Univariate analysis showed that age,education back-ground,employment status,occupational classification,per capita monthly income of families,and the number of comorbidities could directly affect self-management behavior of MHD patients(P<0.05).PLS-SEM model showed that self-efficacy(β = 0.246),family care level(β = 0.124)and social support(β = 0.140)positively impacted patients' self-management behavior,while disease perception negatively affected self-management behavior(β =-0.097).Self-efficacy can mediate the impact of disease perception and social support on self-management behavior of MHD patients(P<0.05).Conclusions Self-management behavior of MHD patients is currently at a low level.Therefore,when devising intervention plans for these patients,it is crucial to consider the combined effects of disease knowledge education,disease perception reconstruction,opportunity provision,and motivation.The primary focus should be on enhancing patients' self-efficacy,which will ultimately elevate their overall level of self-management.

Objective To investigate the effect of ERH gene knockdown on the proliferation and apoptosis of human bladder cancer T24 cells. Methods T24 cells infected by lentivirus with interference on ERH gene sequence were cloned to establish stable T24 cells clone in ERH gene suppression. The expression of ERH mRNA gene in bladder cancer was detected by using quantitative real time polymerase chain reaction (qPCR). The effects of ERH knockout on the cell proliferation and apoptosis were examined by using methylthiazolyl tetrazolium (MTT) assay, colony formation assay and flow cytometry. The effect of ERH knockout on the tumorigenic effect of T24 cells in vivo was verified by subcutaneous tumor formation in nude mice. Results After lentiviral transfection, qPCR results showed that the knockdown effect of ERH mRNA in ERH normal group (untreated T24 cells) was better than that in ERH gene knockdown group, and the difference was statistically significant [(1.006±0.126) vs. (0.079±0.007); t=12.72, P=0.0002]. After knocking out ERH gene, MTT assay showed that the proliferation ability of T24 cells in ERH gene knockdown group was weakened compared with ERH normal group, and the difference was statistically significant [A490 value: (0.13±0.00) vs. (0.66±0.01);t=104.61, P<0.0001]. Colony formation assay indicated that the ability of clone in ERH normal group was weakened compared with ERH gene knockdown group [(10.5 ±1.2) vs. (196.4 ±4.0); t= 73.63, P< 0.0001]. Flow cytometry showed that the cell apoptosis rate in ERH gene knockdown group was higher than that in ERH normal group [(11.0 ±0.5) ％ vs. (4.2 ±0.5) ％; t= 16.06, P<0.0001]. Imaging results of subcutaneous tumor formation in nude mice showed that the total fluorescence intensity of the tumor area in ERH gene knockdown group was (4.67 ±0.59) × 1010 μW/cm2, and the corresponding part in ERH normal group was (9.54±4.20) × 1010μW/cm2 (t=3.64, P=0.0051);tumor weight in ERH gene knockdown group was (0.80±0.62) g, and in ERH normal group was (1.79±0.71) g (t=3.33, P=0.0037). Conclusion ERH gene knockout can inhibit the proliferation of human bladder cancer T24 cells, and promote the cell apoptosis.

Objective:To investigate the effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on targeted therapy of prostate cancer and its effect on tumor microenvironment. Methods:125I-RSOAds-hTERT/PSA ( 125I-virus complex) oncolytic adenovirus was constructed by PCR amplification and double restriction enzyme ligation. TUNEL staining, flow cytometry and Caspase-3 immunoblotting assay were used to detect the killing effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on prostate cancer cells in vitro and in vivo, respectively. To explore the effect of 125I-virus complex on tumor tissue cytokine secretion levels, interleukin 2 (IL-2), IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in the culture supernatant of human prostate cancer cell line PC3, mouse prostate adenocarcinoma cell line RM-1, and mice serum were detected by ELISA. We explored the regulation of 125I-virus complex on the expression of CD24, CD44 and prostate stem cell antigen (PSCA) in prostate tumor tissues and tumor cells through immunohistochemistry. Meanwhile, the expression levels of CD32 and vascular endothelial growth factor (VEGF), as well as CD4+ , CD8+ and macrophage infiltration in tumor tissue were detected through immunofluorescence experiments. Results:125I-virus complex oncolytic adenovirus significantly increased tumor cell apoptosis in vitro and in vivo that was significantly higher than that of 125I group and virus complex group. Meanwhile, IL-2 ( t=-183.30, -38.20, P<0.05), IL-10 ( t=113.80, 92.71, P<0.05), TNF-α ( t=-73.20, -73.91, P<0.05), IFN-γ ( t=-65.37, -139.70, P<0.05) increased in vitro and in vivo. 125I-virus complex reduced the expression of CD24, CD44 and PSCA in tumor cells and tumor tissue, reduced the weight of tumor tissue, inhibited angiogenesis of tumor tissue ( t=8.55, P<0.05), and regulated the immune response in tumor tissue. Conclusions:125I-virus complex targeting prostate cancer can significantly kill cancer cells, reduce the weight and angiogenesis of tumor, and improve tumor microenvironment.

OBJECTIVETo elucidate the current diagnosis and treatment status of gastric cancer in the Chinese population based on three high volume databases.

METHODSClinical and pathological data of patients who underwent gastric cancer resection with complete follow-up information between January 2000 and December 2012 from Sun Yat-sen University Cancer Center, The First Affiliated Hospital of China Medical University and Tianjin Medical University Cancer Hospital were retrospectively analyzed. The overall survival rate was calculated by Kaplan-Meier method. The prognostic risk factors were analyzed by using Cox proportional hazards model.

RESULTSA total of 8 338 cases were enrolled into the study, including 2 977 cases (35.7%) from Sun Yat-sen University Cancer Center, 3 043 cases (36.5%) from The First Affiliated Hospital of China Medical University and 2 318 cases (27.8%) from Tianjin Medical University Cancer Hospital. There were 5 852 male cases and 2 486 female cases with a ratio of 2.4 to 1.0. The age of patients was from 15 to 89 years old (median 59 years old). The ratio of early gastric cancer (T1NanyM0) was 11.5% (956/8 338). There were 2 226 gastric cancer cases (26.7%) originating from the fundus and cardiac region, 1 637 cases (19.6%) from the body, 3739 (44.9%) cases from the antrum, and 736 cases (8.8%) from the whole stomach. The median maximal tumor diameter was (4.5 ± 2.8) cm. Based on the Lauren classification, 3 448 cases (41.4%) were intestinal type and 4 890 cases (58.6%) diffuse type. A total of 1 975 cases (23.7%) and 6 363 cases (76.3%) underwent complete and subtotal gastrectomy respectively. The majority of patients (7 707 cases, 92.4%) underwent radical gastric resection, while 631 cases (7.6%) palliative resection. According to AJCC/UICC seventh edition of gastric cancer TNM staging system, 802 patients (9.6%) were stage I(A, 735 patients (8.8%) stage I(B, 695 patients (8.3%) stage II(A, 1 507 patients (18.1%) stage II(B, 1 247 patients (15.0%) stage III(A, 1 342 patients (16.1%) stage III(B, 1 583 patients (19.0%) stage III (C and 427 patients (5.1%) stage IIII(. The average number of retrieved lymph node was 21.0 ± 13.1, in which 5 761 patients (69.1%) had more than 15 retrieved lymph nodes. The overall 1-, 3-, 5- and 10-year survival rates were 83.0%, 56.8%, 49.1% and 43.0% respectively. For patients receiving radical resection, the 1-, 3-, 5- and 10-year survival rates were 84.9%, 59.5%, 51.7% and 45.3% respectively. The overall 5-year survival rates for different stages were as follows: stage I(A 93.8%, stage I(B 80.8%, stage II(A 70.8%, stage II(B 59.6%, stage III(A 44.4%, stage III(B 32.9%, stage III(C 18.9% and stage IIII( 10.2%. Cox regression model showed that age, tumor site, tumor size, Lauren type, T staging, N staging, M staging and number of retrieved lymph nodes were independent factors affecting the prognosis of gastric cancer patients (P=0.000).

CONCLUSIONRetrospective study on these domestic three high volume databases demonstrates the clinical and pathological characteristics of gastric cancer based on Chinese population, which is expected to stand as a ground of basic data for future clinical research.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; Databases, Factual ; Female ; Gastrectomy ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; diagnosis ; surgery ; Survival Rate ; Young Adult