Objective To study the expression and significance of proliferation cell nuclear antigen (PCNA) ,cyclin dependent ki‐nase 4(CDK4) and p16INK4a in nonneoplastic epithelial disorders of vulvar(NNEDV) and explore the correlation of them .Methods Fifty‐two vulvar lesion samples(lesion group) and 52 vulvar skin surrounding the lesions samples(lesion‐adjacent group) were col‐lected from NNEDV patients ,and lesion group include 27 samples with squamous hyperplasia of vulvar(VSH) ,15 samples with li‐chen sclerosus of vulvar(VLS) and 10 samples containing both lesions(mixed‐lesion) .25 vulvar skin samples were collected from patients receiving perineal repair operation as control group .The expression of PCNA ,CDK4 and p16INK4a was detected by immuno‐histochemistry assay .Results The expression of PCNA in lesion group was significant higher than those in lesion‐adjacent group and control group (P< 0 .05) ,while there was no significant differences among VSH ,VLS and mixed lesion ,and there was no sig‐nificant difference between lesion adjacent group and control group (P> 0 .05) .The expression of CDK4 in lesion group and lesion‐adjacent group were significant higher than that in control group (P < 0 .05) ,while there weren′t significant differences among VSH ,VLS and mixed‐lesion (P> 0 .05) ,and there was no significant difference between lesions group and lesion‐adjacent group (P> 0 .05) .The expression of p16INK4a in lesion group was significant lower than those in lesion‐adjacent group and control group (P< 0 .05) ,while there was no significant differences among VSH ,VLS and mixed‐lesion (P> 0 .05) ,and there was no significant difference between lesion‐adjacent group and control group (P> 0 .05) .About PCNA and CDK4 ,there was a positive correlation in lesion group (P< 0 .05) and there were no correlations in other groups (P> 0 .05) .About PCNA and p16INK4a ,there was a negative correlation in lesion group (P< 0 .05) and there were no correlations in other groups (P > 0 .05) .About CDK4 and p16INK4a ,there were no correlations in all groups (P> 0 .05) .Conclusion expression of PCNA ,CDK4 and p16INK4a may be related of abnormal in‐creasing of cell proliferation and acceleration the process of cell cycle ,which may be applied to molecular target for treatment .

Objective To investigate the current situation of knowledge of stroke rehabilitation among Traditional Chinese Medicine (TCM) physicians in 14 tertiary A-level hospitals. Methods A cross section survey was conducted in 14 tertiary A-level hospital in China in May, 2012. A total of 128 TCM physicians completed the questionnaire, which was created according to Medical Care Guideline of Stroke Rehabilitation in China. Results Overall accuracy of stroke rehabilitation basic knowledge was 66.83%, 88.77% of stroke rehabilitation evaluation, 75.12% of stroke rehabilitation treatment. Conclusion The knowledge of stroke rehabilitation is insufficient among TCM physicians in tertiary A-level hospitals, and further training is needed.

OBJECTIVE: To explore the therapeutic mechanism of Liushen Wan (LSW) against colitis-associated colorectal cancer (CAC) by network pharmacology. METHODS: TCMSP, BATMAN-TCM, CNKI, PubMed, Genecards, OMIM, and TTD databases were used to obtain the related targets of LSW and CAC. The common targets of LSW and CAC were obtained using Venny online website. The PPI network was constructed using Cytoscape 3.8.2 to screen the core targets of LSW in the treatment of CAC. GO and KEGG enrichment analysis were conducted using DAVID database. The therapeutic effect of LSW on CAC was evaluated in a C57BL/6J mouse model of AOM/DSS-induced CAC by observing the changes in body weight, disease activity index, colon length, and size and number of the tumor. HE staining and RT-qPCR were used to analyze the effect of LSW on inflammatory mediators. Immunohistochemistry and TUNEL staining were used to evaluate the effect of LSW on the proliferation and apoptosis of AOM/DSS-treated colon tumor cells. Immunohistochemistry and Western blotting were used to detect the effects of LSW on the expression of TLR4 proteins in CAC mice. RESULTS: Network pharmacology analysis identified 69 common targets of LSW and CAC, and 33 hub targets were screened in the PPI network. KEGG pathway enrichment analysis suggested that the effect of LSW on CAC was mediated by the Toll-like receptor signaling pathway. In the mouse model of AOM/DSS-induced CAC, LSW significantly inhibited colitis-associated tumorigenesis, reduced tumor number and tumor load (P < 0.05), obviously improved histopathological changes in the colon, downregulated the mRNA levels of proinflammatory cytokines, and inhibited the proliferation (P < 0.01) and promoted apoptosis of colon tumor cells (P < 0.001). LSW also significantly decreased TLR4 protein expression in the colon tissue (P < 0.05). CONCLUSION LSW can inhibit CAC in mice possibly by regulating the expression of TLR4 to reduce intestinal inflammation, inhibit colon tumor cell proliferation and promote their apoptosis.
Mice ; Animals ; Toll-Like Receptor 4 ; Colitis-Associated Neoplasms ; Network Pharmacology ; Mice, Inbred C57BL ; Colonic Neoplasms/pathology*

Objective : To study the effect of bufalin on the proliferation and apoptosis of human colorectal cancer cell line HCT116，and to explore the role of endoplasmic reticulum stress ( ERS) in this process． Methods : The effect of bufalin on the proliferation of HCT116 cells was determined by CCK-8 assay．After HCT116 cells were treated with different concentrations of bufalin for 48 hours，cell apoptosis was detected by Annexin V / PI assay， and the expression of apoptosis-related proteins Bax and Bcl-2 was detected by Western blot．At the same time，the expression of ERS-related proteins glucose regulated protein 78 ( GRP78) ，phosphorylated protein kinase R like endoplasmic reticulum kinase ( p-PERK) ，eukaryotic translation initiation factor 2 α ( eIF2 α) ，phosphorylated eukaryotic translation initiation factor 2 α (p-eIF2 α) and C / EBP homologous protein ( CHOP) was detected by Western blot．HCT116 cells were divided into control group，bufalin group and combination group (bufalin + 4-phenylbutyric acid) ，and the expression of apoptosis-related proteins Bax and Bcl-2 was observed by Western blot. Results: CCK-8 assay showed that bufalin could inhibit the proliferation of HCT116 cells．Apoptosis assay showed that bufalin could induce apoptosis of HCT116 cells．The results of Western blot showed that bufalin could up-regulate the expression of pro-apoptotic protein Bax and down-regulate the expression of anti-apoptotic protein Bcl-2．It could also induce ERS and activate PERK / eIF2 α/ CHOP pathway．When bufalin combined with 4-phenylbutyric acid，the apoptosis-promoting effect of bufalin was inhibited． Conclusion Bufalin can effectively inhibit the prolif- erative activity and induce apoptosis of HCT116，which is achieved to some extent by activating ERS.

Objective : To investigate the effects of linoleic acid ( LA) on the diversity and structure of intestinal flora in mice． Methods : Twelve SPF grade male C57BL /6J mice at 7 weeks were randomly divided into control group ( CTRL group) and linoleic acid group (LA group) ．One day before the linoleic acid diet was supplemented， the normal food was removed from the LA group and the mice in the LA group were fasted for one night，so that the LA diet was more acceptable to the mice in the LA group，and LA was given on the day of the experiment recording， and the feed was updated at any time to ensure that the mice could eat freely until the end of modeling．After 12 weeks of modeling，mouse feces were collected，and mixed samples were collected for every two mice feces，and then 16sRNA high-throughput sequencing was performed to analyze intestinal flora structure，Alpha and Beta diversity. Results : 16sRNA high-throughput sequencing showed that LA intervention damaged the richness and diversity of intestinal flora．The results of principal component analysis showed that the composition of flora in CTRL group was different from that in LA group．At gate level，the relative abundance of Actinobacteria in LA group increased (P < 0. 01) ．At the genus level，the relative abundance of L．Duchennei in the LA group decreased (P＜0. 05) ，but the relative abundance of Bifidobacterium，Faecalibaculum and Erysipelotrichaceae in the LA group increased ( all P < 0. 01) ． Conclusion LA intervention could reduce the richness and diversity of intestinal flora in mice，and adjust the structure of intestinal flora．There were significant differences between beneficial bacteria and pathogenic bacte- ria in intestinal flora after LA intervention，which provided certain basis for the treatment of bioactive compounds of linoleic acid and the therapeutic adjustment of intestinal microorganisms as targets.