Objective Galangin is a natural flavonoid with antineoplastic activity .SIRT1 is an important member of Sirtuin family which parcitipate in many physiological process .The aim of this study was to investigate the effect of SIRT 1 on HepG2 cell apop-tosis induced by galangin . Methods HepG2 cells were pre-treated with SIRT1 inhibitor EX-527 for 2 hours, and then galangin for 24 hours.DMSO solvent control group, EX-527 treatment group, galangin treatment group and EX-527 and galangin co-treatment group were established.Hoechst 33342 staining, flow cytometry and western blot were performed to detect the apoptosis of HepG2 cells.After regu-lating the expression of SIRT1 in HepG2 cells with RNA interference and transfection of exogenous genes , these cells were treated with ga-langin for 24 hours.Negative control group , vector control group , SIRT1 knock down group , blank control group , blank vector group ,and SIRT1 upregulation group were established .Western blot and Flow cytometry were performed to detect the apoptosis of HepG 2 cells. Results The apoptosis rate and the gray level ratio of shear band of PARP 1 and GAPDH that of galangin group [(11.62± 0.55) %, 0.89±0.01]and EX-527+galangin group[(25.75±0.61) %, 1.15±0.06] were all increased(P<0.01),when these were compared with DMSO solvent control group [(2.49±0.22) %, 0.06±0.00];and those in EX-527+galangin group were also markedly increased compared with galangin group (P<0.01).The result of western bolt was that the gray level ratio of PARP 1 and GAPDH of SIRT1 knocked down group(0.06±0.01) was markedly decreased compared with vector control group (1.11±0.05)and without adenovi-rus infection group (1.10±0.04)(P<0.01).The apoptosis rate and the gray level ratio of shear band of PARP 1 and GAPDH that of SIRT1 knocked down group were markedly increased compared with vector control group and without adenovirus infection group ( P<0.01).The gray level ratio of PARP1 and GAPDH of SIRT1 up-regulated group (1.63±0.04) was markedly increased compared with blank control group (0.89±0.02) and without plasmid transfection group (0.87±0.03) (P<0.01).The apoptosis rate and the gray lev-el ratio of shear band of PARP 1 and GAPDH that of SIRT1 up-regulated group were markedly decreased compared with blank control group and without plasmid transfection group (P<0.01). Conclusion SIRT1 inhibited galangin-induced apoptosis in HepG2 cells.

Objective Galangin has various antitumor activities and digital holographic microscopy ( DHM ) .This study aimed to investigate the detection of galangin-induced apoptosis of HepG2 cells using DHM. Methods HepG2 cells were cultured in vitro and treated with galangin at 130 μmol/L for 6, 12, and 24 hours respectively to induce apoptosis.Then, MTT assay was used to detect the proliferation of the HepG2 cells, Hoechst 33342 staining and flow cytometry adopted to determine their apoptosis, and DHM employed to visualize their morphological changes. Results Compared with the blank controls, galangin significantly inhibited the proliferation of the HepG2 cells ([7.32 ±2.47]%vs [39.14 ±2.04]%, P<0.01) and obviously induced their nuclear condensa-tion and apoptosis, with a total apoptosis rate of (11.550 ±0.043)%at 24 hours.DHM showed statistically significant differences be-tween the galangin-treated and control groups in the morphological changes of the HepG2 cells at the three time points (P<0.05). Conclusion Digital holographic microscopy can be used to detect the morphological changes of HepG2 cells during galangin-induced apop-tosis.

Objective To investigate the effects of saturated fatty acids and unsaturated fatty acids on proliferation and autophagy of human lung cancer cells. Methods The lung cancer cells A549 were treated with stearic acid (saturated fatty acid) and doconexent (DHA, unsaturated fatty acid), respectively, in concentrations of 0, 30, 60, 120 and 240μmol/L. MTT test and cell clone formation assay were performed to detect the proliferation of A549 cells. The morphology of A549 autophagy was observed by confocal laser scanning microscopy after A549 cells were treated with stearic acid or DHA for 24 hours. Western blotting assay was used to detect the expression of autophagy-related protein after A549 cells were treated with stearic acid or DHA for 12, 24 and 36 hours, respectively. Results 30-240μmol/L stearic acid or DHA both inhibited the proliferation of A549 cells (P<0.05). Both stearic acid and DHA induced autophagy of A549 cells, meanwhile, down-regulated Phospho-mTOR (ser2481) and up-regulated LC3Ⅱ/LC3Ⅰ of A549 cells (P<0.05). Conclusions Both saturated fatty acid and unsaturated fatty acid can inhibit the proliferation and induce autophagy of lung cancer cells. The mechanisms of autophagy may be related to Phospho-mTOR (ser2481) signaling pathway.

Objective To investigate the relationship of plasma homocysteine with the genotype distribution of MTHFR and MTRR among Chinese Han women in Xiangtan. Methods MTHFR C677T, A1298C and MTRR A66G geno?typing was analyzed to detect the distribution of gene polymorphisms among 1 701 women from Xiangtan city then the data were compared with the rest of the Han women in Zibo, Zhengzhou, Yantai, Zhenjiang, Songzi, Huizhou, Qionghai. Plasma Hcy levels from 110 patients were measured and analyzed the correlation with gene polymorphisms. Results The frequency of MTHFR C677T genotype and allele frequencies in Xiangtan is 12.6%which is higher than Huizhou (10.9%) and Qionghai (6.1%) but lower than Zibo (43.6%), Zhengzhou (36.8%), Yantai (32.2%), Zhenjiang (21.8%) with statistically significant dif?ference (P<0.05). There is no significant different in MTHFR C677T between Xiangtan and Songzi. The frequency of MTH?FR A1298C genotype and allele frequencies in Xiangtan is 4.8%which is lower than Qionghai(7.1%)but higher than Zibo (1.4%),Zhengzhou(2.4%), Yantai(1.8%), Zhenjiang(3.5%)and Songzi(2.6%)with statistically significant difference. The frenquency of MTRR A66G genotype and allele frequencies in Xiangtan is 6.8%which is higher than Zibo (4.8%) but lower than Qionghai (9.3%) with statistically signifcant difference. Plasma Hcy concentration correlate with MTHFR C677T, Hcy concentration in TT population is higher than that in CT and CC population(μmol/L:8.52±2.01 vs 5.94±1.47 vs 5.71± 0.18);Plasma Hcy concentration also correlate with MTHFR A1298C and Hcy concentration in CC population is higher than AA and AC population(μmol/L:9.83 ± 2.26 vs 6.35 ± 2.13 vs 5.55 ± 1.75);Plasma Hcy concentration does not correlate with MTRR A66G. Conclusion The gene polymorphism of MTHFR C677T, A1298C and MTRR A66G among the Han women in Xiangtan was statistically different from other selected regions of China. Mutation in MTHFR C 677T and A1298C were associated with elevated plasma levels of Hcy.

Objective To improve the reliability and accuracy of WBC counting in cerebrospinal fluid (CSF) ,this article is stud-ying the improved method of WBC counting in CSF by finding out the optimum percentage of CSF specimen with the most suitable concentration of acetic acid .Methods CSF specimen was mixed with different acetic acid at different ratio respectively .WBC counts were performed in 5 minutes on diluted samples of various concentrations .A series of 20 CSF specimens were analyzed via the proposed assay and conventional method .The average value and coefficient of variation (CV) of WBC count of each sample were c compared and analyzed .Results The optimum percentage of CSF sample was obtained at 60∶40 ratio .In this percentage , the maximal WBC count (189/μL) was obtained compared that of conventional method (161/μL) .Moreover ,the CV of the WBC counts in this percentage (7% ) was also lower than that of the conventional method (18% ) .Conclusion The reliability and accur-ancy of WBC counting in CSF was the optimum percentage of CSF specimen and 5% acetic acid was 60 :40 .It may lead to a more reliable ,accurate and standard way of WBC counts in CSF .

Objective To observe the clinical effect of replantation and non-finger replantation in the treatment of complete distal segment finger amputations,and to analyze the related factors affecting the survival rate of replantation of amputated finger,so as to provide an objective reference for clinical treatment. Methods From March 2015 to June 2016,sixty-two patients with complete distal segment finger amputations treated in the Third People's Hospital of Huizhou were randomly divided into two groups: the observation group and the control group according to the random number table method. The observation group was treated with finger replantation (Pancreatic repair,orthotopic suture and stump remodeling); the two groups of patients with early finger survival rate,replanting fingernail growth,two points discrimination,distal fingertips The clinical data of 288 patients with replantation of single finger rupture were analyzed retrospectively. The clinical data were divided into two groups:the survivors group,the survivors group 74 cases,compared the clinical data of the two groups of patients,analysis of the impact of replantation of the survival rate of the relevant factors. Results The survival rate of early finger was 8. 57% ( 3/ 35) in the observation group and 15. 15% ( 5/ 33) in the control group, the difference was not statistically significant (χ2 = 0. 216,P>0. 05). At 6 months after operation,the length of nail growth,the two-point discrimination and the distal interphalangeal mobility were significantly better than those in the control group((13. 5±2. 9)mmvs. (11. 8±2. 2)mm);(4. 6±0. 3)mmvs. (7. 5±0. 6)mm;(62. 5±4. 4)°vs. (45. 3±3. 6)°) (P<0. 05) . After 6 months,the observation group The excellent and good rate of finger joint activity was 93. 55% (29/ 31). The excellent rate of joint activity was 70. 97% (22/ 31) in the control group,the difference was statistically significant (χ2 = 3. 979,P<0. 05) 288 cases of replantation of clinical data found that smoking history,type of injury,warm ischemic time,degree of disruption,cross-section thrombosis,postoperative skin temperature,pulp elasticity and postoperative psychological status can affect the replantation ( P< 0. 05) . Multivariate logistic regression analysis showed that the factors affecting the survival rate of replantation of finger injury were significantly higher than those of severe depression(OR5. 698,95%CI:2. 892- 8. 738,P< 0. 001)>complete disconnection(OR5. 389,95%CI:2. 672-7. 964,P<0. 001) >warm ischemic time more than 6 hour (OR4. 515,95%CI:1. 366-8. 847,P<0. 001)>postoperative section Thrombosis 成(OR3. 287,95%CI:2. 543~9. 678,P< 0. 001) > low postoperative skin temperature ( OR2. 142,95% CI:1. 243 - 5. 212,P < 0. 001) > poor postoperative knee elasticity(OR2. 008,95% CI:1. 117- 5.449,P< 0. 001) . Conclusion In the treatment of patients with complete injury from the end of the finger,the effect of replantation of the finger is significantly improved,which can improve the postoperative joint activity and improve the appearance and function of the finger. The period of severe depression,complete disconnection and warm ischemia is long Affect the survival rate of finger replantation of the main risk factors for the above factors targeted measures can improve the survival rate of finger replantation.

Objective: To investigate the effect of lncRNA SBF2-AS1 on epithelial-mesenchymal transition (EMT) of cervical cancer HeLa cell via regulating miR-140-5p/VEGFA (vascular endothelial growth factor A) axis. Methods: After cell culture and transfection, the cells were divided into 5 groups: NC group, miR-140-5p mimic group, miR-140-5p mimic+pcDNA-VEGFA group, si-lncRNA SBF2-AS1+pcDNA-VEGFA group and si-lncRNA SBF2-AS1+miR-140-5p mimic group. The expression level of lncRNA SBF2-AS1 in cervical cancer tissues and cell lines was detected by qPCR. The targeted relationship between lncRNA SBF2-AS1, miR-140-5p and VEGFA was confirmed by Dual luciferase reporter gene assay. The expression levels of VEGFA and EMT-related proteins N-cadherin, Vimentin and E-cadherin in HeLa cells were detected by WB. The invasion and migration of HeLa cells were detected by Transwell. Results: lncRNA SBF2-AS1 was highly expressed in cervical cancer tissues and cell lines (P<0.05 or P<0.01). Dual luciferase reporter gene assay confirmed that lncRNASBF2-AS1 targetedly combined with miR-140-5p and VEGFAwas a target gene of miR-140-5p (P< 0.05). Knockdown of lncRNA SBF2-AS1 inhibited invasion and migration as well as EMT of HeLa cells. Further experiment confirmed that lncRNA SBF2-AS1 up-regulated the expression level of VEGFA via miR-140-5p, thereby promoting invasion, migration and EMT of HeLa cells. Conclusion: lncRNASBF2-AS1 promotes EMT of HeLa cells via miR-140-5p/VEGFAaxis.

Objective: To investigate the role of CD11b agonist leukadherin-1 (LA1) in the development of intestinal inflammation and colitis disease in a mouse model of dextran sulfate sodium (DSS)-induced colitis. Methods: The mouse model of experimental colitis was induced by DSS. Body weight changes and survival status were monitored every day. The length of colons was measured at day 7. Colon tissue sections were stained with hematoxylin and eosin (HE) and observed under an optical microscope for pathological analysis. The percentages of apoptotic cells in colon tissues were observed by TUNEL staining. Myeloperoxidase (MPO) activity was measured with MPO activity detection kit. IL-1β and TNF-α levels were detected by ELISA. Macrophages and TNF-α in colon tissues were observed using immunofluorescence staining and confocal microscopy. Flow cytometry was performed to detect the changes in TLR4 expression on macrophages after stimulating mice with LA1 for 0, 3, 6 and 12 h. Moreover, TLR4 expression was also measured by Western blot after treating bone marrow-derived macrophages (BMDMs) with LA1 for 0, 3, 6 and 12 h. Unpaired t-test was used for statistical analysis. Results: Compared with the DSS group, the LA1+ DSS group presented lower mortality rate, greater body weight and longer colon and the differences between the two groups were statistically significant. Moreover, the LA1+ DSS group showed lighter pathological damages, decreased percentage of apoptotic cells and suppressed MPO activity as compared with those of the DSS group. The number of macrophages and the relative concentrations of IL-1β and TNF-α in colon tissues were lower in the LA1+ DSS group than in the DSS group, and the differences between the two groups were statistically significant. There was no significant difference in the total expression of TLR4 on macrophages before and after LA1 treatment. However, the mean flourscence indensity (MFI) of TLR4 was weaker on the LA1-treated macrophages than on the untreated macrophages. Conclusions LA1 could alleviate the DSS-induced experimental colitis in mice through suppressing the activation of TLR4 pathway on macrophages. This study provided a new insight and theoretical reference for understanding the pathogenesis of inflammatory bowel diseases.