Objective To investigate the effects of Xiaotan Sanjie Recipe (XTSJR) on the proliferation and apoptosis of colorectal cancer cell;To study its relevant mechanism. Methods The original XTSJR aqueous solution was lyophilized, weighted, and dissolved in cell culture and stored. HCT116 cell line was treated with different concentrations of XTSJR for 72 h, and the cell proliferation was detected by CCK-8 assay. XTSJR treated HCT116 cell line with the concentration of 0.94 mg/mL and 1.88 mg/mL for 24 h, 48 h, 72 h, and the cell proliferation was detected by CCK-8 assay. HCT116 cell line was treated with different concentrations (0.24, 0.47, 0.94, 1.88 mg/mL) of XTSJR for 72 h. The cell apoptosis was detected by TUNEL assay under the fluorescence microscope, and the post-intervention expression of Caspase3 protein was detected by Western blot. Results The inhibitory effects of XTSJR on the proliferation of HCT116 cell line was concentration-time dependent. The IC50 value at 72 h-time point was 0.94 mg/mL. Medicine concentration and the treated time have interactive contribution to the inhibitory effect. XTSJR induced the cell apoptosis. The apoptosis rate increased with the increasing medicine concentration. There was statistically significant difference compared with the control group ( P<0.01). After treatment the expression of Pro-Caspase3 decreased, while the expression Cleaved Caspase3 protein increased. Conclusion XTSJR inhibits the proliferation of HCT116 cell line and induce apoptosis. Its mechanism might be related to the activity of Caspase3 protein.

Aim To investigate the method of establishing the atopic dermatitis mice model induced by calcipotriol ( MC903 ) . Methods Induction dose exploratory experiment: since d 0, 0.33,1,3 nmol MC903 was smeared on the right ear of BALB/c mice in each dose group respectively , for 7 consecutive days . The ear swelling degree of the mice was observed every day and the bilateral ears thicknesses were measured .The materials were drawn and analyzed in d 3 and d 7.Induction days exploratory experiment: 2 nmol MC903 was smeared on the right ear of BALB/c mice, for 14 consecutive days .The ear swelling degree of mice was observed every day and the bilateral ears thicknesses were measured .The histopathological examination of the right ear was conducted in 3 d, 7 d, 11 d and 15 d respectively .The tissue homogenate of the mice right ear was prepared .The ex-pressions of thymic stromal lymphopoietin (TSLP),IL-33, IL-4 and IFN-γin the homogenate , CD40 +,CD86 +,the DC surface markers and ILC2 contents in the peripheral lymph nodes were detected.Results ①1,3 nmol MC903 induced ear swelling in mice was significantly increased in d 3, the levels of TSLP and IL-4 were significantly increased .The level of IL-33 in 3 nmol dose group was increased significantly in d 7.② The right ear swelling of 2 nmol MC903 induced atopic dermatitis mice was significant , the ear thickness was increased gradually and reached the peak in d 14.The histopathological examination of the mice ear tissue showed in d 7, the right ear tissue of the mice was swelling and red , the capillary vessels were dilated and the infiltration of inflammatory cells was obvious .The ear in-flammatory symptoms maintained and gradually aggravated for 15 days.Compared with the normal mice , MC903 increased the TSLP level in the right ear tissue homogenate significantly in d 3 and then decreased gradually .The levels of IL-4 and IL-33 were increased significantly in d 7 and then decreased gradually .The levels of ILC2, CD40 +, CD86 + in the peripheral lymph nodes were increased in d 7 and d 15.Conclusion The atopic derma-titis mice model can be successfully established using 2 nmol MC903 induced mice for 7 days.Appropriate testing point of TSLP is d 3.Appropriate testing point of IL-33 and IL-4 is d 7.

Objective To study the effects of diet and physical activity factors on blood pressure in nine provinces, using the multilevel model. Methods Data was collected in the China Health and Nutrition Survey (CHNS). A total of 6706 men and 7140 women aged above 18 who attended at least one of the surveys in the year of 1997,2000,2004 and 2006 were selected, and a two-level male and female random intercept-slope growth models were applied to estimate the relationship between the intake of daily salt, vegetable, fruit, fat, protein as well as the time spent on the moderate or heavy physical activity and blood pressure. Results After controlling for age,education, BMI, drinks and total energy intake, mean of the daily salt intake per person was positively associated with systolic blood pressure in women ( β= 0.0481, s-x= 0.0178, P＜0.01 ). Mean of the daily vegetable intake per person was negatively associated with systolic blood pressure in men and women, with the regression coefficients as -0.0063,-0.0068 respectively, indicating that if mean of the daily vegetable intake per person increased by 100 g, the systolic blood pressure would decrease by 0.6 mm Hg (1 mm Hg=0.133 kPa) or more. In addition, the daily vegetable intake was also negatively associated with diastolic blood pressure (P＜0.01). Daily fruit intake was negatively associated with systolic blood pressure both in men and women, with regression coefficients as -0.0029 and -0.0031 respectively. Time spent on the moderate or heavy physical activity was also negatively associated with systolic blood pressure in both men and women, and diastolic blood pressure in women (P＜0.05). No relationship was found between daily fat, protein intake and blood pressure. Conclusion Daily salt, vegetable, fruit intake, time spent on the moderate or heavy physical activity were associated with blood pressure in both men and women. Programs on integrated lifestyle modifications including dietary salt reduction, eating more vegetable and fruits, increasing physical activity level, plus weight control were critical for the control of high blood pressure.

Objective To explore the setting of logos on tobacco control information at outlets for retails and restaurants in 12 selected cities of China.Methods For all the shops for retail of tobacco,alcohol,food and restaurants under survey in 333 blocks of 12 cities (Beijing,Tianjin,Shanghai,Qingdao,Hangzhou,Shaoxing,Suzhou,Nantong,Zhenjiang,Chengdu,Xining and Harbin),setting and contents of logos on tobacco control information,inside and outside them were examined.Results 45 700 objectives were included in the study.Among all types of retail shops,the identification rate of tobacco control information at the entrance and inside were 3.6％ and 4.4％,with an overall identification rate as 7.0％.The overall rate at the entrance of all the restaurants was 4.6％ which was larger than the ones at the retail shops.Our result showed that there were differences between cities and types of establishments and higher rates seen in the larger ones.Of all the places that having had placement of information on tobacco control,only 18.5％ of them had put them both inside and outside.Slogans or images on "No Smoking" were the main forms of information but less than 10％ of them would show signs as ‘exclusive non-smoking'.Conclusion Data from our survey showed that the identification rate of tobacco control information was at a low level in 12 cities,and differences were seen between cities,size of establishment,that called for improvement of the existing tobacco control policies in China.

OBJECTIVETo investigate the expression and the subcellular localization of HDAC9 in different brain regions of mice after cerebral ischemic injury and explore the association between HDAC9 and ischemic stroke.

METHODSTwenty-one male C57BL/6 mice were randomly divided into sham-operated group (n=9) and operated group (n=12). In the latter group, the mice with Zea-Longa neurological deficit scores of 2 or 3 following middle cerebral artery occlusion (MCAO) were assigned into MCAO group (n=9). Immunofluorescence was performed to investigate the subcellular localization of HDAC9 in the brain tissues on day 3 after MCAO. Western blotting and qRT-PCR were used to analyze the expression of HDAC9 in different regions of the brain. Results Immunofluorescence showed more intense HDAC9 expressions in the brain tissues around the infarct focus, and in the cells surrounding the infarct, HDAC9 expression was obviously increased in the cytoplasm and reduced in the cell nuclei. Compared with the other brain regions, the ipsilesional cortex with MCAO showed more abundant HDAC9 expressions at both the mRNA and protein levels (P<0.05).

CONCLUSIONHDAC9 may be closely related to cerebral ischemic injury and participate in the pathophysiological process of ischemic stroke.

BACKGROUND: Repair of tendon interface is a difficulty in orthopedics and sports medicine, and the formation of new bone is conducive to its healing. Calcitonin gene-related peptide (CGRP) plays a critical role in tissue homeostasis and repair, but its effect on the bone-tendon interface repair is unknown. OBJECTIVE: To investigate the effect of CGRP on the expression of osteocalcin and to evaluate the effect of CGRP on the early healing of rotator cuff injury in a mouse model. METHODS: A mouse model of supraspinatus insertion-humerus injury was created. All model mice were then randomized into two groups, and given the injection of 5 nmol/kg CGRP (experimental group) or same volume of normal saline (control group) through the glenohumeral joint immediately after operation, thrice weekly, for 2 weeks. The mice were sacrificed at postoperative 4 and 6 weeks to remove the rotator cuff samples for hematoxylin-eosin staining and biomechanical test. The mRNA and protein expression levels of osteocalcin were surveyed by qRT-PCR, immunohistochemistry, and western blot assay. RESULTS AND CONCLUSION: At postoperative 4 and 6 weeks, the mRNA and protein expression of osteocalcin in the experimental group were significantly higher than those in the control group (P < 0.05). Compared with the control group, there were more fibrocartilages in the experimental group at 4 weeks postoperatively, and more new bone formation in the experimental group at 6 weeks postoperatively. At 4 weeks postoperatively, failure load in the experimental group increased slightly, but it was not significantly different from that in the control group (P > 0.05); at 6 weeks postoperatively, failure load in the experimental group was significantly higher than that in the control group (P<0.05). To conclude, local injection of CGRP can up-regulate the expression of osteocalcin and the formation of new bone at the injury site, which can enhance early healing the injured rotator cuff.

OBJECTIVE: To investigate the expression of fibroblast growth factor receptor 2 (FGFR2) in clear cell renal cell carcinoma (ccRCC; or kidney renal clear cell carcinoma, KIRC), to analyze the relationship between the expression of FGFR2 and the clinical pathological features and prognosis of ccRCC, to study the relationship between the expression of FGFR2 and other molecules, and to explore its role in the development of ccRCC. METHODS: Gene expressional and clinical information of ccRCC patients were downloaded from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus(GEO) database. Next, the data were transformed and collated. In the study, 104 clinical ccRCC samples and corresponding paracancerous normal tissue samples were collected from Department of Urology, Peking University First Hospital. Immunohistochemistry (IHC) was performed and the staining results were scored, so as to compare the expression of FGFR2 in ccRCC and paracancerous normal tissues. Besides, quantify real-time polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression level of FGFR2 in normal renal epithelial cell lines (293) and ccRCC cell lines (786-O, 769-P, OSRC-2, Caki-1, ACHN, and A498). In addition, the relationship between FGFR2 expression and clinical pathological characteristics (including TNM staging and pathological grading) and survival prognosis in ccRCC patients was further analyzed. Furthermore, the relationship between FGFR2 expression and B cells, T cells, natural killer (NK) cells and neutrophil infiltration in the ccRCC patients was analyzed, and the Biological General Repository for Interactionh Datasets (BioGRID) was used to builds protein-protein interaction (PPI) networks to study molecules that interacted with the FGFR2 protein. RESULTS: In the TCGA database, the expression of FGFR2 was down-regulated in ccRCC tissue samples compared with normal tissue samples, and the expression in the GEO database also showed this differences. Furthermore, FGFR2 expression was downregulated in ccRCC clinical samples and ccRCC cell lines, compared with corresponding paracancerous normal tissue or normal renal epithelial cell lines. In addition, FGFR2 high expression was associated with earlier, lower-level ccRCC and was associated with a better prognosis in the patients with ccRCC. Moreover, FGFR2 expression was not significantly related to B cells, T cells, NK cells and neutrophil infiltration, and the PPI network showed that FGFR2 protein interacted with certain molecules. CONCLUSION Our work sheds light on the potential role of FGFR2 in the development of ccRCC, suggesting that FGFR2 may serve as a prognostic marker and potential therapeutic target for patients with ccRCC.
Biomarkers, Tumor/analysis* ; Carcinoma, Renal Cell/pathology* ; Humans ; Kidney Neoplasms/pathology* ; Prognosis ; Receptor, Fibroblast Growth Factor, Type 2/genetics*

Background/Aims: Ubiquitination is widely involved in the progression of hepatocellular carcinoma (HCC) by regulating various cellular processes. However, systematic strategies for screening core ubiquitin-related genes, clarifying their functions and mechanisms, and ultimately developing potential therapeutics for patients with HCC are still lacking. Methods: Cox and LASSO regression analyses were performed to construct a ubiquitin-related gene prediction model for HCC. Loss- and gain-of-function studies, transcriptomic and metabolomics analysis were used to explore the function and mechanism of UBE2S on HCC cell glycolysis and growth. Results: Based on 1,423 ubiquitin-related genes, a four-gene signature was successfully constructed to evaluate the prognosis of patients with HCC. UBE2S was identified in this signature with the potential to predict the survival of patients with HCC. E2F2 transcriptionally upregulated UBE2S expression by directly binding to its promoter. UBE2S positively regulated glycolysis in a HIF-1α-dependent manner, thus promoting the proliferation of HCC cells. Mechanistically, UBE2S enhanced K11-linkage polyubiquitination at lysine residues 171 and 196 of VHL independent of E3 ligase, thereby indirectly stabilizing HIF-1α protein levels by mediating the degradation of VHL by the proteasome. In particular, the combination of cephalomannine, a small molecule compound that inhibits the expression of UBE2S, and PX-478, an inhibitor of HIF-1α, significantly improved the anti-tumor efficacy. Conclusions UBE2S is identified as a key biomarker in HCC among the thousands of ubiquitin-related genes and promotes glycolysis by E3 enzyme-independent ubiquitination, thus serving as a therapeutic target for the treatment of HCC.

Background/Aims: Ubiquitination is widely involved in the progression of hepatocellular carcinoma (HCC) by regulating various cellular processes. However, systematic strategies for screening core ubiquitin-related genes, clarifying their functions and mechanisms, and ultimately developing potential therapeutics for patients with HCC are still lacking. Methods: Cox and LASSO regression analyses were performed to construct a ubiquitin-related gene prediction model for HCC. Loss- and gain-of-function studies, transcriptomic and metabolomics analysis were used to explore the function and mechanism of UBE2S on HCC cell glycolysis and growth. Results: Based on 1,423 ubiquitin-related genes, a four-gene signature was successfully constructed to evaluate the prognosis of patients with HCC. UBE2S was identified in this signature with the potential to predict the survival of patients with HCC. E2F2 transcriptionally upregulated UBE2S expression by directly binding to its promoter. UBE2S positively regulated glycolysis in a HIF-1α-dependent manner, thus promoting the proliferation of HCC cells. Mechanistically, UBE2S enhanced K11-linkage polyubiquitination at lysine residues 171 and 196 of VHL independent of E3 ligase, thereby indirectly stabilizing HIF-1α protein levels by mediating the degradation of VHL by the proteasome. In particular, the combination of cephalomannine, a small molecule compound that inhibits the expression of UBE2S, and PX-478, an inhibitor of HIF-1α, significantly improved the anti-tumor efficacy. Conclusions UBE2S is identified as a key biomarker in HCC among the thousands of ubiquitin-related genes and promotes glycolysis by E3 enzyme-independent ubiquitination, thus serving as a therapeutic target for the treatment of HCC.

Background/Aims: Ubiquitination is widely involved in the progression of hepatocellular carcinoma (HCC) by regulating various cellular processes. However, systematic strategies for screening core ubiquitin-related genes, clarifying their functions and mechanisms, and ultimately developing potential therapeutics for patients with HCC are still lacking. Methods: Cox and LASSO regression analyses were performed to construct a ubiquitin-related gene prediction model for HCC. Loss- and gain-of-function studies, transcriptomic and metabolomics analysis were used to explore the function and mechanism of UBE2S on HCC cell glycolysis and growth. Results: Based on 1,423 ubiquitin-related genes, a four-gene signature was successfully constructed to evaluate the prognosis of patients with HCC. UBE2S was identified in this signature with the potential to predict the survival of patients with HCC. E2F2 transcriptionally upregulated UBE2S expression by directly binding to its promoter. UBE2S positively regulated glycolysis in a HIF-1α-dependent manner, thus promoting the proliferation of HCC cells. Mechanistically, UBE2S enhanced K11-linkage polyubiquitination at lysine residues 171 and 196 of VHL independent of E3 ligase, thereby indirectly stabilizing HIF-1α protein levels by mediating the degradation of VHL by the proteasome. In particular, the combination of cephalomannine, a small molecule compound that inhibits the expression of UBE2S, and PX-478, an inhibitor of HIF-1α, significantly improved the anti-tumor efficacy. Conclusions UBE2S is identified as a key biomarker in HCC among the thousands of ubiquitin-related genes and promotes glycolysis by E3 enzyme-independent ubiquitination, thus serving as a therapeutic target for the treatment of HCC.