tiRNA-Val promotes angiogenesis via Sirt1-Hif-1α axis in mice with diabetic retinopathy

BACKGROUND: Diabetic retinopathy (DR) is a specific microvascular complication arising from diabetes, and its pathogenesis is not completely understood. tRNA-derived stress-induced RNAs (tiRNAs), a new type of small noncoding RNA generated by specific cleavage of tRNAs, has become a promising target for several diseases. However, the regulatory function of tiRNAs in DR and its detailed mechanism remain unknown. RESULTS: Here, we analyzed the tiRNA profiles of normal and DR retinal tissues. The expression level of tiRNA-Val was significantly upregulated in DR retinal tissues. Consistently, tiRNA-Val was upregulated in human retinal microvascular endothelial cells (HRMECs) under high glucose conditions. The overexpression of tiRNA-Val enhanced cell proliferation and inhibited cell apoptosis in HRMECs, but the knockdown of tiRNA-Val decreased cell proliferation and promoted cell apoptosis. Mechanistically, tiRNA-Val, derived from mature tRNA-Val with Ang cleavage, decreased Sirt1 expression level by interacting with sirt1 3'UTR, leading to the accumulation of Hif-1α, a key target for DR. In addition, subretinal injection of adeno-associated virus to knock down tiRNA-Val in DR mice ameliorated the symptoms of DR. CONCLUSION: tiRNA-Val enhance cell proliferation and inhibited cell apoptosis via Sirt1/Hif-1α pathway in HRMECs of DR retinal tissues.

Effect of Geniposide on Liver Injury and Bile Acid Profile in Serum, Liver and Excrement of Rats / 中国实验方剂学杂志

Objective:To investigate the hepatotoxicity of different doses of geniposide on the liver of rats and the effects on bile acid profile in serum， liver tissue and feces. Method:The 60 Sprague Dawley rats， half male and half female， were randomly divided into 5 groups according to body weight： blank group and four different doses （50， 100， 200， 400 mg·kg^-1） geniposide groups， 12 rats in each group. The rats were treated by gavage once a day for 7 consecutive days， and the serum， liver and cecal contents were collected on the 8^th day of treatment. The activities of alanine aminotransferase （ALT）， aspartate aminotransferase （AST） and alkaline phosphatase （ALP）， the contents of albumin （ALB）， total bilirubin （TBIL）， total bile acid （TBA）， creatinine （Crea） and carbamide （Urea） were detected in each group. The sections of liver tissue were stained with hematoxylin-eosin（HE）， and the protein expressions of cytokeratin 7（CK7） and cytokeratin 19（CK19） were detected by immunohistochemistry. The protein expressions of CK7 and CK19 in the liver tissue were detected by Western blot. And the mRNA expressions of cholesterol 7<italic>α</italic>-hydroxylase （CYP7A1）， cholesterol 27<italic>α</italic>-hydroxylase （ CYP27A1） and cholesterol 12<italic>α</italic>-hydroxylase （CYP8B1） were detected by real-time PCR. The contents of 18 kinds of bile acids in serum， liver and cecal contents were determined by ultra-performance liquid chromatography-mass spectrometry（UPLC-MS）. Result:Compared with the control group， TBIL level in each dose of geniposide group was increasesd significantly （<italic>P</italic><0.01）. ALT， AST activity and TBA content in 400 mg·kg^-1 geniposide group were increased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. HE staining showed that， compared with control group， there was bile duct reaction in the portal area and inflammatory cells infiltrate around bile duct in 200 mg·kg^-1 and 400 mg·kg^-1 geniposide groups， especially 400 mg·kg^-1. The expressions of CK7 and CK19 in liver tissue of 400 mg·kg^-1 geniposide group were significantly higher than those in the control group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the control group， the contents of glycoursodeoxycholic acid （GUDCA） and glycohyodeoxycholic acid （GHDCA） in liver tissue of 400 mg·kg^-1 geniposide group decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）， the contents of sodium taurochenodeoxycholate （TCDCA）， hyodeoxycholic acid （HDCA）， cholic acid （CA） and chenodeoxycholic acid （CDCA） in liver tissue increased significantly （<italic>P</italic><0.01）， the contents of glycocholic acid hydrate （GCA）， glycochenodeoxycholic acid （GCDCA）， glycodeoxycholic acid hydrate （GDCA）， glycocholic acid （GLCA）， tauroursodeoxycholic acid （TUDCA）， GUDCA， GHDCA， ursodeoxycholic （UDCA） and taurolithocholic acid （TLCA） decreased， the proportions of TCDCA， HDCA， CA， CDCA and deoxycholic acid （DCA） in liver tissue increased， the contents of GHDCA and lithocholic acid （LCA） in serum decreased significantly （<italic>P</italic><0.01）， while sodium taurohyodeoxycholate hydrate （THDCA）， taurocholic acid （TCA）， GCA， TCDCA， UDCA， CA， CDCA， DCA in serum decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. The contents of CA， UDCA， CA， CDCA and DCA increased significantly （<italic>P</italic><0.05）， the ratio of CA/DCA increased significantly （<italic>P</italic><0.05）， and the ratio of CA and CDCA increased by 19.60% and 4.63%， respectively； Compared with the control group， the contents of all bile acids in cecal contents of 400 mg·kg^-1 were decreased， and the contents of GCA， UDCA， HDCA， GCDCA， GDCA， TLCA， GLCA， CDCA， DCA and LCA were decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. In addition， real-time PCR results showed that the mRNA expressions of CYP7A1， CYP27A1 in the 400 mg·kg^-1 geniposide group were significantly higher than those in the control group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:The 400 mg·kg^-1geniposide can cause obvious hepatotoxicity in rats， and the bile acid profile in liver， serum and excrement changes significantly， and the changes of the each bile acid in liver， serum and feces are different. However， the causal relationship between the gardenoside-induced liver injury and the changes in bile acid profile are<italic> </italic>not clear. It needs to be further studied.

Effect of Xiayuxue Tang with Different Processing Method on CCl4-induced Hepatic Fibrosis in Rats / 中国实验方剂学杂志

Objective::To compare the anti-hepatic fibrosis effect of ethanol extract of honey Xiayuxue Tang (H-Xiayuxue Tang)and prepared slices of crude drug-Xiayuxue Tang (C-Xiayuxue Tang). Method::Totally 40 wistar rats were randomlydivided into normal group, model group, H-Xiayuxue Tang group, C-Xiayuxue Tang group, and Fuzheng Huayu group. Liver fibrosis model was induced by hypodermic injection of 50% carbon tetrachloride for 9 weeks. From the seventh week, the normal group and the model group were given 0.3% sodium carboxymethyl cellulose by gavage. The other three groups were given H-Xiayuxue Tang(0.58 g·kg-1), C-Xiayuxue Tang (0.43 g·kg-1), Fuzheng Huayu (2 g·kg-1), respectively. At the end of the ninth week, liver tissues and serum samples were collected for follow-up studies. The pathological changes and fibrosis degree were observed by HE and Sirius red staining, hydroxyproline content in liver tissues was detected by alkaline hydrolysis, serum liver function was determined by automatic analysis, the expressions of collagen type I (COL-I) and α-smooth muscle actin (α-SMA) in liver tissues were detected by Real-time PCR. The expressions of α-SMA in liver tissues was detected by Western blot and Immunohistochemistry, and serum inflammatory factors and oxidantion indexes were determined by ELISA, respectively. Result::Compared with control group, the severity of liver fibrosis was observed, the area ratio of Sirius red positive staining, hydroxyproline (Hyp) content, the protein and mRNA expressions of α-SMA, COL-I in the liver tissue were significantly increased in the model group (P<0.01). And the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBil), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-17A (IL-17A) and malondialdehyde (MDA) in the serum of the rats were markedly increased (P<0.01). The levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were apparently decreased (P<0.01). Compared with model group, the degree of liver fibrosis was significantly improved, the area ratio of Sirius red positive staining, the content of Hyp, the protein and mRNA expression of α-SMA and COL-I, and the serum levels of AST, ALT, TBil, TNF-α, IL-1β, IL-17A and MDA were significantly decreased (P<0.05, P<0.01). The serum contents of SOD and GSH-Px were markedly increased (P<0.05, P<0.01), respectively, by H-Xiayuxue Tang or C-Xiayuxue Tang. Among them, the improvement of liver histopathological changes was more obvious in the H-Xiayuxue Tang group than in the C-Xiayuxue Tang group. Meanwhile, the percentage of Sirius red positive staining area, the content of hydroxyproline, the expressions of COL-I and α-SMA mRNA and protein in liver tissue were significantly decreased compared with the C-Xiayuxue Tang group(P<0.05, P<0.01). In addition, there was no significant difference in serum liver function, inflammatory factors and oxidantion related indicators between groups, but the improvement trend of H-Xiayuxue Tang was more obvious. Conclusion::H-Xiayuxue Tang and C-Xiayuxue Tang have a better effect in hepatic fibrosis, especially H-Xiayuxue Tang, suggesting that honey can increase the anti-hepatic fibrosis effect of Xiayuxue Tang.

Effect of electrical stimulation at auricular points combined with sound masking on the expression of CREB, BDNF and TrkB in the auditory cortex of tinnitus rats / 中国针灸

OBJECTIVE@#To explore the effect of electrical stimulation at auricular points (EAS) combined with sound masking on the expression of cAMP-response element binding protein (CREB), brain-derived neurotrophic factor (BDNF) and tyrosine receptor kinase B (TrkB) in the auditory cortex of tinnitus rats.@*METHODS@#A total of 27 adult male SD rats were randomly divided into a control group, a model group and an EAS group. The rats in the model group and the EAS group were intervened with intraperitoneal injection of sodium salicylate to induce tinnitus model, while the rats in the control group were intervened with injection of 0.9% NaCl solution. After the model was successfully established, the rats in the EAS group were treated with electrical stimulation at "Shenmen" (TF) and "Yidan" (CO), combined with sound masking; the treatment was given once a day for 15 days. The gap prepulse inhibition of acoustic startle (GPIAS) and prepulse inhibition (PPI) testing were performed using the acoustic startle reflex starter package for rats. The expression of BDNF, TrkB, CREB and p-CREB in the auditory cortex of each group were measured with Western Blot analysis.@*RESULTS@#① Compared with the control group, the GPIAS values in 12 kHz, 16 kHz, 20 kHz and 28 kHz were significantly decreased in the model group (all 0.05).@*CONCLUSION@#EAS could improve the GPIAS values of high-frequency background sound in tinnitus rats, which may be related with the upregulation of the BDNF/TrkB/CREB signaling pathway in the auditory cortex, leading to the reversion of the maladaptive plasticity.

Analysis of research status and hotspots of snail intestinal flora based on bibliometrics / 中国血吸虫病防治杂志

Objective To understand the current research focus and trends in the field of snail intestinal flora. Methods The literature focusing on snail intestinal flora and published from 1998 to 2017 were retrieved from the core database of Web of Science. The quantitative analysis of literature was then conducted by using CiteSpace software based on the bibliometricsmethod.The research trends were then summarized systematically, and the potential research fronts and focuses were explored. Results Totally 139 articles were identified in the field of snail intestinal flora. The top three countries with highest publications included the United States of American, Brazil, and South Korea; while the top three institutions were Kyung Hee University, Osvaldo Cruz Foundation, and Oxford University. Five terms were identified as the key words in this field, including diversity, cellulose, Achatina fulica, lignocelluloses, and species nova. Meanwhile, 5 critical papers with the citation frequency over 15 were recognized, and 5 study clusters were formed including the application, diversity, and function of intestinal flora, difference of snail source and flora, and newly discovered bacteria in the snail intestine. Conclusion The current research focuses on intestinal flora of snails include the diversity, function and application of intestinal flora.

Analysis of research status and hotspots of snail intestinal flora based on bibliometrics / 中国血吸虫病防治杂志

Objective To understand the current research focus and trends in the field of snail intestinal flora. Methods The literature focusing on snail intestinal flora and published from 1998 to 2017 were retrieved from the core database of Web of Science. The quantitative analysis of literature was then conducted by using CiteSpace software based on the bibliometricsmethod.The research trends were then summarized systematically, and the potential research fronts and focuses were explored. Results Totally 139 articles were identified in the field of snail intestinal flora. The top three countries with highest publications included the United States of American, Brazil, and South Korea; while the top three institutions were Kyung Hee University, Osvaldo Cruz Foundation, and Oxford University. Five terms were identified as the key words in this field, including diversity, cellulose, Achatina fulica, lignocelluloses, and species nova. Meanwhile, 5 critical papers with the citation frequency over 15 were recognized, and 5 study clusters were formed including the application, diversity, and function of intestinal flora, difference of snail source and flora, and newly discovered bacteria in the snail intestine. Conclusion The current research focuses on intestinal flora of snails include the diversity, function and application of intestinal flora.

Mechanism of quercetin improving rat coronary artery myogenic re-sponse under high glucose / 中国病理生理杂志

AIM:To investigate the mechanism of quercetin improving rat coronary artery myogenic response under high glucose ( HG) by measuring muscle tension of coronary arterial ring and recording voltage -gated K +channel ( Kv) current of coronary artery smooth muscle cells by whole cell patch clamp .METHODS:The coronary rings from the normal SD rats were acutely isolated , and then divided into 6 groups:(1) control group;(2) HG group;(3) HG+low dose (3 μmol/L) of quercetin group;(4) HG+moderate dose (10 μmol/L) of quercetin group; (5) HG+high dose (30 μmol/L) of quercetin group;(6) HG+C6303 (PKC inhibitor) +high dose of quercetin group.Determinations of coronary artery response to vasoconstrictor (60 mmol/L KCl or 0.1 mmol/L U46619) or vasodilator (Ach at 10 -9 ~10 -5 mol/L) were performed, and the percentage of coronary ring tension was calculated using the contraction as 100%caused by 60 mmol/L KCl.The rat coronary artery smooth muscle cells were acutely isolated for recording the Kv current using whole cell patch clamp .RESULTS:Compared with control group , the contraction amplitudes to 60 mmol/L KCl or 0.1 mmol/L U46619 were significantly increased under HG incubation .Quercetin intervention concentration-dependently re-duced the coronary artery contraction amplitude .Incubation of PKC specific inhibitor C 6303 attenuated the effect of querce-tin.Compared with control group , the diastolic amplitude to Ach decreased significantly in HG group , and quercetin inter-vention concentration-dependently increased the coronary artery diastolic amplitude .Incubation of PKC specific inhibitor C6303 attenuated the effect of quercetin .Compared with control group , HG incubation inhibited Kv current of coronary ar-tery vascular smooth muscle cells significantly , and quercetin intervention attenuated the inhibitory effect of HG on Kv cur-rent intensity .Incubation of PKC specific inhibitor C 6303 attenuated the effect of quercetin .CONCLUSION: Quercetin has a protective effect on myogenic response of coronary artery under HG and the effects is related to the increase in Kv cur -rent and the activation of PKC in vascular smooth muscle cells .

Mechanism of quercetin improving rat coronary artery myogenic re-sponse under high glucose / 中国病理生理杂志

AIM:To investigate the mechanism of quercetin improving rat coronary artery myogenic response under high glucose ( HG) by measuring muscle tension of coronary arterial ring and recording voltage -gated K +channel ( Kv) current of coronary artery smooth muscle cells by whole cell patch clamp .METHODS:The coronary rings from the normal SD rats were acutely isolated , and then divided into 6 groups:(1) control group;(2) HG group;(3) HG+low dose (3 μmol/L) of quercetin group;(4) HG+moderate dose (10 μmol/L) of quercetin group; (5) HG+high dose (30 μmol/L) of quercetin group;(6) HG+C6303 (PKC inhibitor) +high dose of quercetin group.Determinations of coronary artery response to vasoconstrictor (60 mmol/L KCl or 0.1 mmol/L U46619) or vasodilator (Ach at 10 -9 ~10 -5 mol/L) were performed, and the percentage of coronary ring tension was calculated using the contraction as 100%caused by 60 mmol/L KCl.The rat coronary artery smooth muscle cells were acutely isolated for recording the Kv current using whole cell patch clamp .RESULTS:Compared with control group , the contraction amplitudes to 60 mmol/L KCl or 0.1 mmol/L U46619 were significantly increased under HG incubation .Quercetin intervention concentration-dependently re-duced the coronary artery contraction amplitude .Incubation of PKC specific inhibitor C 6303 attenuated the effect of querce-tin.Compared with control group , the diastolic amplitude to Ach decreased significantly in HG group , and quercetin inter-vention concentration-dependently increased the coronary artery diastolic amplitude .Incubation of PKC specific inhibitor C6303 attenuated the effect of quercetin .Compared with control group , HG incubation inhibited Kv current of coronary ar-tery vascular smooth muscle cells significantly , and quercetin intervention attenuated the inhibitory effect of HG on Kv cur-rent intensity .Incubation of PKC specific inhibitor C 6303 attenuated the effect of quercetin .CONCLUSION: Quercetin has a protective effect on myogenic response of coronary artery under HG and the effects is related to the increase in Kv cur -rent and the activation of PKC in vascular smooth muscle cells .

Overexpression of amphiregulin promotes the growth of mouse colon carcinoma CT26 cells in vivo / 肿瘤

Objective: To investigate the effect of epidermal growth factoramphiregulin (AREG) on the growth of mouse colon carcinoma CT26cells and its related mechanisms.Methods: The protein expression level of AREG in different mouse cancer cells was detected by ELISA. Mouse colon carcinoma CT26 cells, melanoma B16 cellsand hepatocellular carcinoma LPC-Akt cells were transfected with the recombinant lentiviralplasmid carrying AREG gene, while the ones transfected with empty plasmid were used asthe negative controls. After AREG overexpression, the cell proliferation, colony-formingabilities and cell cycle progression in vitro were detected by MTT, colony-forming assay andFCM, respectively. After the homograft mouse model of CT26 cells was constructed, thegrowth of homograft tumor was observed, the distribution of immune cells in tumor tissueswas detected by FCM, furthermore the expression of chemokine was detected by real-timefluorescent quantitative PCR.Results: The levels of AREG expression were relatively low in mouse colon carcinoma CT26cells, melanoma B16 cells and hepatoma LPC-Akt cells. AREG overexpression did notmarkedly affect the proliferation, colony-forming abilities and cell cycle progression of thesethree types of tumor cells in vitro (all P > 0.05). However, in the homograft mouse model ofCT26 cells, AREG overexpression significantly promoted the growth of tumor cells in vivo (P <0.01), decreased the percentage of CD8+ T cells (P < 0.05), and reduced the mRNA level ofCC chemokine 5 ligand (CCL5) (P < 0.05) which was related to CD8+ T cell recruitment.Conclusion: AREG promotes the growth of mouse colon carcinoma CT26 cells in vivo . AREGmay affect the tumor microenvironment by regulating the production of chemokine which isrelated to CD8+ T cell recruitment.