Aim To iuvestigate the effect and the mechanism of salmonmilt DNA (SMD) on age-related involutions in mouse thymus. MethodsFemale BALB/c of 10 months were divided randomly into three groupsaccording to their weights: high dosage group 333.33 mg/(kg @ d), lowdosage group 166. 67 mg/(kg @ d) and control group 0 mg/(kg @ d) .After five weeks, with Image-Pro Plus (version. 4.0) software, the thymusindexes and the thymoctytes in the thymus section were measured, as wellas the thymus cortex thickness. All the data were analyzed by SAS statisticsoftware. Mieroarray technique was applied to screen the gene fragments,which were differently expressed between the high dosage group and thecontrol group, together with RT-PCR to further confirm some of them.Results No significant differences of the variables including bodyweight, thymus weight and thymus indexes among the three groups werefound (F ＜ 3.0 and P ＞ 0.05, respectively). The thymocytes quanti-ties of thymus cortex and medulla in the high dosage group were significantlyhigher than those of the control group [cortex D(H, C) = 9.46, P ＜ 0.01;medulla t( H.C) = 2.53, P ＜ 0.05]. The thymus cortex thicknesses of bothSMD supplement groups were significantly higher than that of the control group[cortex D(L,C)=3.65, P＞ 0.05; medulla t(L, C)=0.8, P＞ 0.05] .112differently expressed gene fragments were isolated. Furthermore, we foundthe fragments with the logged number of U23789, X80232 and Aw209102were highly expressed in the high dosage group when RT-PCR techniquewas used. Conclusion SMD may reverse the age-related involutions inmouse thymus via up-regulation the expression of proliferation related genesand development and differentiation related genes simultaneously.

Objective:To investigate the current situation in construction and operation status of official WeChat public platform of community hospitals in Shanghai Downtown and provide the reference for effective application of WeChat public platform in community hospitals.Methods:The current situation of health service provided with WeChat public platform of community hospitals was investigated by website survey.We focused on the opening rate,menu services,information push and off-line operations of WeChat public platform from the perspective of ethics.Results:Of the investigated 98 hospitals,a total of 48 WeChat public platforms were established,accounting for 49.0％.Among which 28 public platforms provided menu bar service,accounting for 58.3％.The public platforms still needed to be improved in terms of article push quantity,reading quantity and daily management.Conclusion:The development level of WeChat public platform of community hospital in shanghai downtown is uneven and WeChat public platform of community hospital exists low service level,imperfect management,lack of publicity and other problems generally.It is recommended that hospitals strengthen the construction of WeChat platform from three aspects,including strengthening team management,keeping the "Six in One" and seeking for commercial assistance.

Objective To investigate the effect of human colorectal fibroblast(CCD-18Co)-conditioned medium(CCD18-Co-CM)on biological behaviors of colorectal cancer(CRC)cells and explore the possible molecular mechanisms.Methods Real-time cellular analysis(RTCA),clone formation assay and wound healing assay were used to analyze the changes in proliferation,clone formation,and migration abilities of CRC cell lines HCT116 and Caco-2 treated with CCD18-Co-CM.Western blotting was used to detect the changes in ATK,ERK and STAT3 signaling pathways in the CRC cells activated by CCD18-Co-CM.The effect of CCD18-Co-CM on spheroidization ability of the cells was assessed with sphere-formation assay,and the changes in expressions of CRC stemness markers were detected using RT-PCR.Results CCD-18Co-CM significantly promoted proliferation,colony formation,and migration of HCT116 and Caco-2 cells,enhanced sphere-forming ability and expressions of CRC stemness markers,and increased ERK phosphorylation in the cells.Treatment with SCH772984 effectively inhibited CCD-18Co-CM-induced ERK signaling pathway activation,suppressed the malignant phenotype,and lowered the sphere-forming ability and expression of stemness markers of the two CRC cells.Conclusion Colorectal fibroblasts promote malignant phenotype of CRC cells by activating the ERK signaling pathway.

Objective To investigate the effect of human colorectal fibroblast(CCD-18Co)-conditioned medium(CCD18-Co-CM)on biological behaviors of colorectal cancer(CRC)cells and explore the possible molecular mechanisms.Methods Real-time cellular analysis(RTCA),clone formation assay and wound healing assay were used to analyze the changes in proliferation,clone formation,and migration abilities of CRC cell lines HCT116 and Caco-2 treated with CCD18-Co-CM.Western blotting was used to detect the changes in ATK,ERK and STAT3 signaling pathways in the CRC cells activated by CCD18-Co-CM.The effect of CCD18-Co-CM on spheroidization ability of the cells was assessed with sphere-formation assay,and the changes in expressions of CRC stemness markers were detected using RT-PCR.Results CCD-18Co-CM significantly promoted proliferation,colony formation,and migration of HCT116 and Caco-2 cells,enhanced sphere-forming ability and expressions of CRC stemness markers,and increased ERK phosphorylation in the cells.Treatment with SCH772984 effectively inhibited CCD-18Co-CM-induced ERK signaling pathway activation,suppressed the malignant phenotype,and lowered the sphere-forming ability and expression of stemness markers of the two CRC cells.Conclusion Colorectal fibroblasts promote malignant phenotype of CRC cells by activating the ERK signaling pathway.

AIM:To explore the molecular mechanism by which leukemia inhibitory factor(LIF)enhances the stemness characteristics of colorectal cancer(CRC)cells by regulating the expression of the molecular marker CD44 in cancer stem cells(CSCs).METHODS:The expression of LIF in CRC tissues was analyzed using the TCGA public data-base and RNAscope in situ hybridization method.Stable knockdown of LIF was constructed in CRC cell lines(HCT116 and Caco2 cells)using a lentiviral infection system.The experiment was divided into 4 groups:CRC cells control group,CRC cells plus LIF group,CRC cells knockdown control group,and CRC cells knockdown LIF group.The impact of exog-enous LIF on CRC cells' capacity for spheroidization was assessed through stem cell spheroidization assays.The ability of CRC cells to proliferate and migrate was evaluated using MTT,RTCA,plate cloning,and migration tests following the ad-dition or removal of LIF.The impact of LIF on the expression of the transcription factor ELF3 and CD44 in CRC tumor cells was determined using RT-qPCR,a fully automated protein expression analysis system,and Western blot.RT-qPCR was used to identify changes in CD44 splicing variant expression following LIF knockdown.RESULTS:The results dem-onstrated that the expression level of LIF in CRC tissue was higher than that in neighboring cancer tissue(P<0.01),and that patients with CRC who had high expression of LIF had a shorter disease-free survival time(P<0.05).While LIF knockdown can decrease CRC cells' capability to proliferate and migrate(P<0.05),exogenous LIF can boost CRC cells' sphericity,proliferation,and migration capacity(P<0.05).Exogenous LIF can raise(P<0.05)CD44 expression in CRC tumor cells,however LIF reduction can lower(P<0.01)CD44 expression and lower the transcription level of CD44 splic-ing variants.Exogenous injection of LIF or LIF knockdown can cause a significant change(P<0.05)in the expression lev-el of the transcription factor ELF3.CONCLUSION:LIF improves the stemness characteristics of colorectal cancer cells by upregulating the transcription factor ELF3,which also promotes the development of malignancy by increasing CD44 ex-pression in CRC cells.

OBJECTIVE: To investigate the mechanism by which conditioned medium of colorectal cancer cells promotes the formation of cancer-associated fibroblasts (CAFs). METHODS: Normal human colorectal fibroblasts (CCD-18Co cells) in logarithmic growth phase were treated with the conditioned media of colorectal cancer HCT116 cells (HCT116-CM) or Caco-2 cells (Caco-2-CM) alone or in combination with 300 nmol/L ERK inhibitor SCH772984. The expression levels of CAFs-related molecular markers were detected in the treated cells with real-time quantitative PCR (RT- qPCR) and immunofluorescence assay, and the changes in cell proliferation, colony formation and migration were assessed with RTCA, colony formation and wound healing assays; Western blotting was performed to detect the activated signaling pathways in the fibroblasts and the changes in CAFs formation after blocking of the signaling pathway. RESULTS: HCT116-CM and Caco-2-CM significantly upregulated mRNA expression levels of CAFs markers (including α-SMA, FAP, FN and TGF-β) in CCD-18Co cells, and strongly promoted fibroblast transformation into CAFs (P < 0.05). The two conditioned media also promoted the proliferation, colony formation and migration of CCD-18Co cells (P < 0.05) and significantly increased the levels of α-SMA protein and ERK phosphorylation in the cells (P < 0.05). The ERK inhibitor SCH772984 obviously inhibited the expression of α-SMA and the transformation of CCD-18Co cells into CAFs induced by the conditioned medium of colorectal cancer cells (P < 0.05). CONCLUSION Colorectal cancer cells may induce the formation of colorectal CAFs by activating the ERK pathway in the fibroblasts.
Humans ; Cancer-Associated Fibroblasts/metabolism* ; Culture Media, Conditioned/pharmacology* ; MAP Kinase Signaling System ; Caco-2 Cells ; Fibroblasts ; Signal Transduction ; Cell Proliferation ; Cell Line, Tumor ; Colorectal Neoplasms/genetics* ; Cell Movement

Objective:To investigate the effects of ectodysplasin-A1 (EDA1) on the proliferation and cell cycle of ameloblast-like epithelial cells (LS8 cells).Methods:Wild EDA1 plasmid pCR3-Flag-EDA1-W (wild group), syndrome mutant EDA1 plasmid pCR3-Flag-EDA1-H252L (mutant group) and empty vector plasmid pCR3-Flag (control group) were transfected into LS8 cells. Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay and cell cycle was detected by flow cytometry. All tests were repeated three times.Results:Compared with the control group (0.105±0.032), the proliferation activity of the wild group (0.201±0.009) was significantly higher after 72 h ( P<0.05). Compared with the control group (0.168±0.054) and the mutant group (0.194±0.059), the proliferation activity of the wild group (0.386±0.066) was significantly higher after 96 h ( P<0.05). There was no significant difference between the mutant group and the control group at all time points ( P>0.05). In the G 0/G 1 phase, compared with the control group (65.4%±2.1%) and the mutant group (66.6%±3.1%), the cell distribution ratio of the wild group (51.2%±1.1%) was significantly lower ( P<0.01). In the S phase, compared with the control group (23.1%±2.0%) and the mutant group (21.9%±1.8%), the cell distribution ratio of the wild type group (37.3%±2.4%) was significantly higher ( P<0.01). There was no significant difference in cell cycle distribution between the mutant group and the control group ( P<0.05). Conclusions:Wild EDA1 promotes the proliferation of LS8 cells and the transformation from G 0/G 1 to S phase. The syndrome mutant EDA1 (EDA1-H252L) loses its function of regulating the cell proliferation and cell cycle of LS8 cells.