Objective:To discuss the role of angiopoietin-like protein 6(ANGPTL6)in liver fibrosis,and to analyze the improving effect of engineered exosome(Exo)-delivered ANGPTL6 mRNA on liver fibrosis.Methods:A total of 12 C57BL/6 mice were randomly divided into olive oil group(OIL group)(intraperitoneally injected with olive oil)and carbon tetrachloride(CCl4)group(intraperitoneally injected with a mixture of olive oil and CCl?),with 6 mice in each group;another 12 C57BL/6 mice were randomly divided into control group(fed a with methionine-choline sufficient diet)and methionine-choline deficient(MCD)group(fed a with MCD diet),and two kinds of mouse liver fibrosis models were established.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting method were used to detect the ANGPTL6 mRNA and protein expression levels in liver tissue of the mice in various groups.A total of 30 mice were randomly divided into olive oil+phosphate buffered saline(PBS)group(OIL+PBS group)(intraperitoneally injected with olive oil twice a week for 8 weeks,then injected with PBS buffer by tail vein twice a week for 6 weeks),CCl4+Exo-green fluorescent protein(GFP)mRNA group(established liver fibrosis model by intraperitoneal injection of CCl4 mixture and were injected by tail vein with engineered Exo loaded with GFP mRNA for 6 weeks),and CCl?+Exo-ANGPTL6 mRNA group(established liver fibrosis model by intraperitoneal injection of CCl4 mixture and were injected by tail vein with engineered Exo loaded with ANGPTL6 mRNA for 6 weeks),with 10 mice in each group.The mice in CCl4+Exo-GFP mRNA group and CCl4+Exo-ANGPTL6 mRNA group were injected with engineered Exo twice a week,20 μg per mouse each time(volume 100 μL).ELISA method was used to detect the serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)activities in the mice in various groups;Masson staining and Sirius red staining were used to observe the collagen deposition in liver tissue of the mice in various groups;immunohistochemistry method was used to detect the α-smooth muscle actin(α-SMA)expression levels in liver tissue of the mice in various groups;RT-qPCR method was used to detect the expression levels of α-SMA,collagen type Ⅰ alpha 1 chain(Col1a1),transforming growth factor β1(TGF-β1),and tissue inhibitor of metalloproteinase 1(TIMP-1)mRNA in liver tissue of the mice in various groups.Results:The bioinformatics analysis results showed that ANGPTL6 expression was significantly down-regulated in activated hepatic stellate cell(aHSC).The ultrasound examination results showed that the liver surface of the mice in OIL group was fine and smooth;compared with OIL group,the liver section of the mice in CCl? group was rough and uneven.The RT-qPCR and Western blotting results showed that compared with OIL group,the ANGPTL6 mRNA and protein expression levels in liver tissue of the mice in CCl? group were significantly decreased(P<0.05).The engineered Exo extracted from the supernatant of HEK293T cells had intact structure and could be largely enriched in the fibrotic liver after tail vein injection,with GFP protein being largely expressed in the liver.The ELISA assay results showed that compared with OIL+PBS group,the ALT and AST activities in CCl4+Exo-GFP mRNA group were significantly increased(P<0.05);compared with CCl4+Exo-ANGPTL6 mRNA group,the serum ALT and AST activities in CCl4+Exo-GFP mRNA group were significantly decreased(P<0.05).The Masson staining and Sirius red staining results showed that compared with OIL+PBS group,the collagen deposition in liver tissue of the mice in CCl?+Exo-GFP mRNA group was significantly increased,and the relative collagen area was increased(P<0.05);compared with CCl4+Exo-GFP mRNA group,the collagen deposition in tissue liver of the mice in CCl?+Exo-ANGPTL6 mRNA group was significantly decreased,and the relative collagen area was decreased(P<0.05).The immunohistochemistry results showed that compared with OIL+PBS group,the α-SMA protein expression level in liver tissue of the mice in CCl?+Exo-GFP mRNA group was significantly increased(P<0.05);compared with CCl4+Exo-GFP mRNA group,the α-SMA protein expression level in liver tissue of the mice in CCl?+Exo-ANGPTL6 mRNA group was significantly decreased(P<0.05).The RT-qPCR results showed that compared with OIL+PBS group,the expression levels of Col1a1,α-SMA,TGF-β1,and TIMP-1 mRNA in liver tissue of the mice in CCl?+Exo-GFP mRNA group were significantly increased(P<0.05);compared with CCl4+Exo-GFP mRNA group,the expression levels of Col1a1,α-SMA,TGF-β1,and TIMP-1 mRNA in liver tissue of the mice in CCl?+Exo-ANGPTL6 mRNA group were significantly decreased(P<0.05).Conclusion:Engineered Exo-delivered ANGPTL6 mRNA injected via the tail vein in the mice is mainly enriched in the liver,and engineered Exo delivery of ANGPTL6 mRNA has an improving effect on liver fibrosis in the mice.

Objective To explore the impact of heat shock factor 2 (HSF2) on the development of lung cancer by promoting interleukin (IL)-10 expression.Methods 50 lung cancer tissues and adjacent normal tissues selected from 50 patients,the expression level of mRNA and protein of HSF2 and IL-10 were respectively detected by RT-PCR,Western blot and Immunohistochemistry;To interfere with expression of HSF2 in A549 cells by siRNA,the expression level of IL-10 was detected by Western blot.Results Compared with the adjacent normal tissues,HSF2 of 76％ (38 of 50) cases were up-regulated (P＜0.01),IL-10 of 80％ (40 of 50) eases were up-regulated (P＜0.01),protein level consistent with mRNA level.The up-regulation expression of IL-10 in lung cancer tissues and HSF2 positively correlated (R2 =0.921 6).The expression of IL-10 in A549 cells was weakened through interference with HSF2 by siRNA.Conclusion HSF2 may contribute to the development of lung cancer by facilitating the expression of IL-10.

Objective: To investigate the efficacy of Baofukang suppositories on cervical columnar epithelial ectopic and the influence on the expression of uterine tissue intercellular adhesion molecule 1 (ICMI-1), transforming growth factor b1 (TGF-b1) mRNA and inflammatory cytokines in cervical tissues.Methods: Totally 102 patients with cervical columnar epithelial ectopic were randomly divided into the observation group and the control group with 51 ones in each.The observation group received Baofukang suppositories, and the control group was treated with cryosurgery.The clinical efficacy was compared between the groups, and before and after the treatment, the levels of ICMI-1 mRNA, TGF-b1 mRNA and inflammatory cytokines were also recorded and compared.Results: The clinical curative effect of the observation group was 94.12%,which was better than that of the control group (P＜0.05).After the treatment, the levels of ICMI-1 mRNA and TGF-b1 mRNA decreased when compared with those before the treatment (P <0.05), and those in the observation group were significantly lower than those in the control group (P＜0.05).The differences in IL-1 and IL-6 of the control group before and after the treatment were not statistically significant (P >0.05), while those in the observation group decreased after the treatment (P <0.05), which were significantly lower than those in the control group (P＜0.05).There were no significant changes in menstrual period and menstrual cycle in the two groups before and after the treatment, and no adverse reactions occurred during the treatment.Conclusion: Baofukang suppositories are effective in the treatment of columnar epithelial ectopic, which can reduce the levels of ICMI-1 mRNA, TGF-b1 mRNA and serum inflammatory cytokine in the patients.

Objective To assess the left ventricular systolic asynchronicity in chronic ischemic model with real-time three-dimensional echocardiography (RT-3DE), and to explore the affection of low-dose dobutamine to it. Methods A chronic ischemic model was induced by placing an Ameroid constrictor in the left circumflex(LCX) in swines,then full volume RT-3DE was performed by Philips iE33 with X3-1 probe combining rest and stress(dobutamine stress echocardiography, DSE) every week after LCX constriction.Ten normal pigs before operation served as controls (group A). Examination of all the models post operation were grouped into group B (mild stenosis, LCX stenosis＜50% ), group C (moderate stenosis, LCX stenosis 50%～75%) and group D (severe stenosis, LCX stenosis≥75%) according to the results of coronary angiography. Images were copied to QLAB 5.2 postprocess workstation,and 3DQA software was used to analyze the full volume data sets. The time to the point with minimal systolic volume (Tmsv) in each segment was taken to derive the following indexes of systolic synchrony: the maximum difference of Tmsv (Tmsv-dif) and standard deviation(Tmsv-SD) among various segments and standard index (Tmsv-dif% and Tmsv-SD%), to evaluate left ventricular dyssynchrony. Tmsv3-6 represented the maximum difference of Tmsv between lateral segment and posterior septum (Tmsv3-5: between lateral segment and inferior) in basal level. Results Tmsvl2-Dif%, Tmsv6-Dif%, Tmsv3-6% and Tmsv3-5% under stress condition in group C and D were significantly higher than those at rest;all the data in group D were significantly higher than in group A and B, and in group C higher than group A ( P ＜0.05,0.01 ). Compared with group A,Tmsv6-Dif,Tmsv3-6 and Tmsv3-5 in group B were significantly increased under stress condition,and so did their standardize data under both rest and stress conditions ( P ＜ 0.05, 0. 01 ). Conclusions RT-3DEcombined with DSE could display sensitively the left ventricular asynchrony caused by chronic ischemia,and that will be more significant in lateral wall in LCX stenosis than in normal segments.