Objective:To explore the possible anti-atherosclerotic mechanisms of glucose co-transporter-2 inhibitor canagliflozin.Methods:ApoE -/-mice fed on Western diet were randomly assigned into the model group ( n=10) and the canagliflozin group ( n=10). C57BL/6J mice fed on normal diet were chosen as the control group ( n=10). Mice in the canagliflozin group were gavaged with canagliflozin for 14 weeks. The presence and severity of atherosclerosis were evaluated with HE and oil red O stainings in aortic root section slices. PCR assay was performed to determine the mRNA expression levels of nitric oxide synthase. Hepatic transcriptome analysis and hepatic amino acid detection were conducted using RNA-seq and targeted LC-MS, respectively. Results:HE staining and oil red O staining of the aortic root showed that AS models were successfully established in ApoE -/-mice fed on Western diet for 14 weeks. Canagliflozin alleviated the severity of atherosclerosis in pathology. Hepatic transcriptome analysis indicated that canagliflozin impacted on amino acid metabolism, especially arginine synthesis in ApoE -/-mice. Targeted metabolomics analysis of amino acids showed that canagliflozin reduced hepatic levels of L-serine, L-aspartic acid, tyrosine, L-hydroxyproline, and L-citrulline, but raised the hepatic level of L-arginine. Compared to the model group, the canagliflozin group exhibited higher serum arginine and nitric oxide levels as well as elevated nitric oxide mRNA expression in aortic tissues ( P<0.05). Conclusion:Canagliflozin regulated the amino acid metabolism, reduced the levels of glucogenic amino acids,and promoted the synthesis of arginine in atherosclerotic mice.

Objective:To explore the effects of canagliflozin on cardiac function and its regulation of ferroptosis in rats with heart failure with preserved ejection fraction (HFpEF).Methods:Thirty-two 7-week-old Dahl salt-sensitive rats were selected and randomly divided into four groups: the control group (fed with low-salt diet), the HFpEF group (fed with high-salt diet), the canagliflozin 20 group (fed with high-salt diet and 20 mg·kg -1·d -1 canagliflozin), and the canagliflozin 30 group (fed with high-salt diet and 30 mg·kg -1·day -1 canagliflozin). Body weight and blood pressure of the rats in each group were monitored. Metabolic cage tests were conducted at the10 th week of the experiment, and echocardiography was performed at the 12 th week, after which the rats were killed. Blood and left ventricular samples were collected. HE staining, Masson staining, Prussian blue iron staining, and reactive oxygen species staining were performed to observe the cardiomyocyte size and shape, degree of interstitial fibrosis, iron staining, reactive oxygen species production under optical microscope. The ultrastructure of cardiomyocytes was observed under electron microscope. Western blotting and real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-qPCR) were used to detect the expression levels of proteins and mRNA related to ferroptosis in left ventricular myocardial tissue of rats in each group. Results:After 1 week of adaptive feeding, all rats survived. Metabolic cage results showed that compared with control group, rats in the HFpEF group, canagliflozin 20 group and canagliflozin 30 group had more food intake, water intake and urine output, and lower body weight (all P<0.05). These changes were more pronounced in canagliflozin 20 group and canagliflozin 30 group than in HFPEF group, and only the body weight at the 12 th week showed a statistically significant difference between canagliflozin 20 group and canagliflozin 30 group ( P<0.05). The blood pressure of 6 th week and 12 th week, heart weight and left ventricular corrected mass of 12 th week of rats in HFpEF group were higher than those in control group, canagliflozin 20 group and canagliflozin 30 group, while the ratio of early mitral valve peak velocity to late mitral valve peak velocity of 12 th week was lower (all P<0.05). HE and Masson staining showed that compared to control group, the myocardial fibers in the left ventricular myocardial tissue of rats in HFpEF group were disordered, with larger cell diameter ((0.032±0.004) mm vs. (0.023±0.003) mm, P<0.05), irregular shape, obvious proliferation of interstitial collagen fibers, and higher collagen volume fraction (0.168±0.028 vs. 0.118±0.013, P<0.05). Compared with HFpEF group, rats in the canagliflozin 20 group and canagliflozin 30 had more orderly arranged myocardial fibers, more regular cardiomyocyte shape, smaller cell diameter, and lower collagen volume fraction ( P<0.05). It was observed under electron microscopy that, compared to control group, most of the striated muscles in myocardial tissue of HFpEF group were broken, and the Z line and M line could not be clearly distinguished, some changes such as mitochondrial swelling, membrane thickening, cristae reduction or even disappearance occurred. In the canagliflozin 20 group and canagliflozin 30 group, the arrangement of striated muscles in the myocardial tissue of rats tended to be more regular, and the morphological changes of mitochondria were milder. Prussian blue iron staining results showed that the iron content in myocardial tissue of rats in HFpEF group was higher than that in control group, canagliflozin 20 group and canagliflozin 30 group. Reactive oxygen species staining results showed that the reactive oxygen species content in the myocardial tissue of rats in HFpEF group was higher than that of control group, canagliflozin 20 group and canagliflozin 30 group. Biochemical analysis of myocardial tissue showed that Fe 2+ and malondialdehyde content in myocardial tissue of rats in HFpEF group were higher than those in control group, canagliflozin 20 group and canagliflozin 30 group, while glutathione content was lower (all P<0.05). Western blot and RT-qPCR detection results showed that compared to control group, rats in HFpEF group had higher expression levels of transferrin receptor 1 (protein relative expression level: 1.37±0.16 vs. 0.31±0.12), acyl-CoA synthetase long-chain family member 4 (protein relative expression level: 1.31±0.15 vs. 0.63±0.09) protein and mRNA, and lower expression levels of ferritin heavy chain 1 (protein relative expression level: 0.45±0.08 vs. 1.41±0.15) protein and mRNA (all P<0.05). There was no statistically significant difference in these indicators between canagliflozin 20 group and the canagliflozin 30 group (all P>0.05). There was no significant difference in levels of glutathione peroxidase 4 protein and mRNA expression in myocardial tissue of rats in four groups( P>0.05). Conclusion:Canagliflozin improves cardiac function in HFpEF rats by regulating the ferroptosis mechanism.

Objective:To discuss the differences in serum proline dehydrogenase(ProDH)levels among chronic heart failure(CHF)patients with different ejection fraction types,and to clarify the effect of ProDH levels on cardiac function.Methods:A retrospective analysis of clinical data of 118 CHF patients was conducted.These patients were divided into heart failure with reduced ejection fraction(HFrEF)group(n=39),heart failure with mid-range ejection fraction group(HFmrEF)(n=42),and heart failure with preserved ejection fraction(HFpEF)group(n=37).A total of 45 non-CHF patients hospitalized during the same period were collected as control group.The general data of all the subjects in various groups were collected,and the levels of biochemical indicators and cardiac structure indicators in serum of all the subjects were detected.Spearman correlation analysis and point-biserial correlation analysis were used to analyze the correlation between serum ProDH levels and various biochemical indicators;multivariate Logistic regression analysis was used to analyze the factors influencing HFrEF and HFmrEF.Results:Compared with control group,the usage rate of beta-blockers of the patients in HFpEF group was significantly increased(P<0.05);in HFmrEF group,the percentage of male patients,the usage rate of statins,and the usage rate of beta-blockers were all significantly increased(P<0.05);in HFrEF group,the age and systolic blood pressure(SBP)of the patients were significantly decreased(P<0.05),while the usage rates of statins and beta-blockers of the patients were significantly increased(P<0.05).Compared with HFpEF group,the age of the patients in HFmrEF group was significantly decreased(P<0.05),and the percentage of male patients and the usage rate of statins were significantly increased(P<0.05);the age of the patients in the HFrEF group was significantly decreased(P<0.05),and the usage rate of statins was significantly increased(P<0.05).Compared with HFmrEF group,the SBP of the patients in HFrEF group was significantly decreased(P<0.05).Compared with control group,the serum levels of low-density lipoprotein cholesterol(LDL-c)of the patients in HFpEF and HFmrEF groups were significantly decreased(P<0.05),while the levels of N-terminal pro-brain natriuretic peptide(NT-proBNP)were significantly increased(P<0.05);the serum levels of glomerular filtration rate(GFR)and ProDH of the patients in HFrEF group were significantly decreased(P<0.05),and the levels of fasting blood glucose(FBG)and NT-proBNP were significantly increased(P<0.05).Compared with HFpEF group,the serum hemoglobin(Hb)level of the patients in HFmrEF group was significantly increased(P<0.05);the serum NT-proBNP level of the patients in HFrEF group was significantly increased(P<0.05),while the ProDH level was significantly decreased(P<0.05).Compared with HFmrEF group,the serum NT-proBNP level of the patients in HFrEF group was significantly increased(P<0.05).Compared with control group,the left atrial diameter(LAD)and the ratio of early diastolic mitral inflow velocity to early diastolic mitral annular velocity(E/Em)of the patients in HFpEF,HFmrEF,and HFrEF groups were significantly increased(P<0.05);the left ventricular end-diastolic diameter(LVEDD)of the patients in HFmrEF and HFrEF groups were significantly increased(P<0.05),and the left ventricular ejection fraction(LVEF)were significantly decreased(P<0.05).Compared with HFpEF group,the LVEDD of the patients in HFmrEF and HFrEF groups were significantly increased(P<0.05),and the LVEF were significantly decreased(P<0.05);the LAD of the patients In HFrEF group was significantly increased(P<0.05).Compared with HFmrEF group,the E/Em ratio of the patients in HFrEF group was significantly increased(P<0.05),and the LVEF was significantly decreased(P<0.05).The serum ProDH levels of the patients were negatively correlated with LVEDD(r=-0.210,P=0.007)and positively correlated with LVEF(r=0.220,P=0.005).Male and elevated FBG levels were the risk factors for cardiac function,while the increasing serum GFR and ProDH levels were the protective factors for cardiac function.Conclusion:There are differences in ProDH levels among the CHF patients with different ejection fraction types.The patients with poorer cardiac function have lower serum ProDH levels,and higher ProDH levels may be beneficial for improving the left ventricular systolic function in the CHF patients.

Objective To investigate the risk factors for rebleeding after emergency esophageal variceal ligation (EVL) in patients with liver cirrhosis. Methods A retrospective analysis was performed for the clinical and laboratory data of 290 patients with liver cirrhosis who underwent emergency EVL in The Fifth Medical Center of Chinese PLA General Hospital from January 2016 to December 2019, and according to the presence or absence of rebleeding within 1-year follow-up, they were divided into rebleeding group and non-rebleeding group. The t -test was used for comparison of normally distributed continuous data between two groups, and the Wilcoxon rank sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. A multivariate logistic regression analysis was performed with the statistically significant factors as independent variables to screen out the independent risk factors for rebleeding after emergency EVL, and the receiver operating characteristic (ROC) curve was plotted to obtain the indices for predicting the probability of rebleeding and establish a predictive model. Results The univariate analysis showed that there were significant differences between the two groups in platelet count ( t =-1.888, P =0.047), Child-Pugh score ( χ 2 =5.975, P =0.049), albumin level ( t =-2.229, P =0.029), and splenic vein diameter ( t =3.808, P =0.001). The multivariate logistic regression analysis showed that Child-Pugh score (odds ratio [ OR ]=0.280, 95% confidence interval [ CI ]: 0.108-0.729, P =0.009), splenic vein diameter ( OR =1.549, 95% CI : 1.197-2.005, P =0.001) and albumin level ( OR =0.832, 95% CI : 0.729-0.949, P =0.006) were independent influencing factors for rebleeding after EVL. The predictive model based on these three factors had an area under the ROC curve of 0.796, with a sensitivity of 83.7% and a specificity of 74.5% at the cut-off value of -0.086. Conclusion Child-Pugh score, albumin level, and splenic vein diameter are independent risk factors for rebleeding after emergency EVL, and the combination of the three indices has the highest sensitivity and specificity in predicting rebleeding.

Objective:To investigate the expression profile change of long non-coding RNA (IncRNA) and mRNA in plasma samples before and after drug resistance of gefitinib for non-small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR)-sensitive gene mutation treated, and to screen out RNA molecule related to gefitinib-resistance.Methods:A total of 12 NSCLC patients with EGFR-sensitive gene mutation treated by gefitinib from Xianyang Center Hospital of Shaanxi Province and Yongchuan Hospital of Chongqing Medical University from March 2015 to April 2019 were selected. Plasma samples before and after drug resistance were collected, and 6 samples in sensitive stage and 6 samples in drug-resistant stage were taken. Gene microarray was used to screen the differentially expressed lncRNA and mRNA; the biological pathway and the function of the differentially expressed mRNA were obtained by using the gene ontology (GO) function annotation analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis.Results:The microarray detection results showed that the expression profiles of lncRNA and mRNA in the plasma of NSCLC patients were different before and after gefitinib-resistance. Fold change≥2 and P < 0.05 were taken as the differential gene screening standard, finally 38 differentially expressed lncRNAs and 53 differentially expressed mRNAs were found. Compared with the sensitive stage, 18 lncRNAs were differentially up-regulated and 20 lncRNAs were down-regulated in the drug-resistant stage; the largest up-regulation lncRNA was RP1-102K2.6 (fold change was 47.31), and the largest down-regulation lncRNA was RP11-149I2.4 (fold change was 24.34). In mRNA expression microarray, compared with sensitive stage, the expressions of 29 mRNAs were up-regulated and 24 mRNAs were down-regulated in the drug-resistant stage, the largest up-regulation mRNA was CUL2 (fold change was 58.49), the largest down-regulation mRNA was CHEK2 (fold change was 23.29). GO functional analysis showed that the differentially expressed mRNA in the plasma of patients with gefitinib-resistance were enriched in the apoptosis and protein binding regulation process. KEGG analysis showed that the differentially expressed mRNA mainly targeted cancer pathway, NSCLC pathway and other pathways. Conclusion:For NSCLC patients with EGFR gene sensitive mutation, there are multiple differentially expressed lncRNAs and mRNAs in plasma before and after drug resistance, and the differential expression may play an important role in the mechanism of gefitinib resistance.

Objective:To evaluate the clinical effect of the treatment of scar formation after severe acne type Ⅳ by acupuncture.Methods:In Henan Traditional Chinese Medicine Hospital, 60 patients with acne who met the inclusion criteria were selected and divided into observation group and control group according to the randomized controlled grouping method. The observation group(9 male and 21 female, age 18-29) was treated with fire needle and salicylic acid.The control group(10 male and 20 female, age 18-27 ) was treated with needle and salicylic acid.The patients in both groups were treated with tanshinone capsule and salicylate acne pill orally.The course of treatment was 8 weeks. The two groups of clinical efficacy, acne severity score scale (Gags) to evaluate symptom improvement, vancouver scar scale, acne scar weight score were observed and compared.Results:Eight weeks after treatment, the effect rate was 90.00% (27/30) in the observation group and 68.97% (20/29) in the control group. The scores of Vancouver scar scale were (3.66 ± 0.91) in the observation group and (5.06 ± 1.09) in the control group ( P< 0.01). The scores of acne scar weight score were (39.40 ± 16.90) in the observation group and (53.16 ± 17.19) in the control group( P< 0.01). The scores of Gags in the observation group and the control group were (10.31 ± 2.14) and (17.55 ± 2.63)( P< 0.01), respectively. Conclusions:The treatment of Ⅳ grade severe acne skin lesions by fire needle is safe, effective, reliable and easy to operate. It can safely and effectively improve the patient’s facial skin lesions, and reduce the possibility of scar formation and pigmentation, so it is worthy of clinical application.

Objective:To evaluate the clinical effect of the treatment of scar formation after severe acne type Ⅳ by acupuncture.Methods:In Henan Traditional Chinese Medicine Hospital, 60 patients with acne who met the inclusion criteria were selected and divided into observation group and control group according to the randomized controlled grouping method. The observation group(9 male and 21 female, age 18-29) was treated with fire needle and salicylic acid.The control group(10 male and 20 female, age 18-27 ) was treated with needle and salicylic acid.The patients in both groups were treated with tanshinone capsule and salicylate acne pill orally.The course of treatment was 8 weeks. The two groups of clinical efficacy, acne severity score scale (Gags) to evaluate symptom improvement, vancouver scar scale, acne scar weight score were observed and compared.Results:Eight weeks after treatment, the effect rate was 90.00% (27/30) in the observation group and 68.97% (20/29) in the control group. The scores of Vancouver scar scale were (3.66 ± 0.91) in the observation group and (5.06 ± 1.09) in the control group ( P< 0.01). The scores of acne scar weight score were (39.40 ± 16.90) in the observation group and (53.16 ± 17.19) in the control group( P< 0.01). The scores of Gags in the observation group and the control group were (10.31 ± 2.14) and (17.55 ± 2.63)( P< 0.01), respectively. Conclusions:The treatment of Ⅳ grade severe acne skin lesions by fire needle is safe, effective, reliable and easy to operate. It can safely and effectively improve the patient’s facial skin lesions, and reduce the possibility of scar formation and pigmentation, so it is worthy of clinical application.

Objective:To investigate the clinical characteristics, incidence trend, underlying diseases, causative drug and prognosis of drug-induced liver injury (DILI), so as to provide basis for its prevention and treatment.Methods:A retrospective study was conducted on 2 820 DILI cases who were admitted to our hospital from January 2002 to December 2015, and their clinical characteristics, incidence trends, underlying related diseases, causative drug, treatment and outcome were analyzed.Results:Among 2 820 DILI cases, the ratio of male to female was 1:1.44, and the age was (44.00±16.32) years old. According to the clinical classification of DILI, there were 2 353 cases (83.43%) of hepatocyte injury, 353 cases (12.51%) of cholestatic type and 114 cases (4.04%) of mixed type. In the three clinical classification of DILI, there was no statistically significant difference in the ratio of male to female (χ 2 = 3.032, P > 0.05). However, the difference in the ratio of male to female between different age groups was statistically significant (χ 2 = 48.367, P < 0.001). Among the patients with liver disease and acute liver disease admitted to our hospital from January 2002 to December 2015, the proportion of DILI and acute DILI showed an overall upward trend. The main underlying related diseases of 2 820 DILI cases were fever (15.14%), skin diseases (11.84%), cardiovascular and cerebrovascular diseases (11.17%). Chinese herbal patent medicines (37.49%), antibiotics (15.85%), antipyretic-analgesics (14.37%), and so on were the main causative drugs involved, and the prognostic differences among the three clinical classifications of DILI in terms of cure, improvement, ineffectiveness, and death were statistically significant ( H = 61.300, P < 0.001). Conclusion:In recent years, among the patients with liver disease in our hospital, the proportion of DILI has shown an obvious upward trend, involving a variety of underlying diseases and causative drugs, and thus it needs clinical attention.

Objective To explore the expression of the Pendrin gene (SLC26A4) and protein in multinodular goiter.Methods Thyroid tissues were obtained from 40 multinodular goiter patients undergoing surgery while the control group were obtained from 40 nomal thyroid tissues.RT-PCR was used to test SLC26A4 gene while western blot and immunohistochemistry were used to test Pendrin protein expression and distribution.Results SLC26A4 mRNA expression in multinodular goiter tissue was significantly increased in comparison with normal nodular tissues (t=2.663,P=0.011).Pendrin protein expression in multinodular goiter group was higher than that in normal tissue (t=2.286,P=0.026).The immunohistochemistry results showed that the Pendrin protein in multinodular goiter was mainly located in cytoplasm.There was positive expression in 24 patients (60％) in multinodular goiter group,while it was in 14 patients (35％) in the normal control group.The difference was significant (X2=5.013,P=0.025).Pendrin protein mainly expressed in cytoplasm in multinodular goiter tissue while it was mainly in cytomembrane in the normal control group.Conclusion SLC26A4 mRNA and its coding protein Pendrin expression are increased in multinodular goiter group,and mainly located in cytoplasm,indicating that iodide transporter function may be damaged when multinodular goiter occurs.

Objective To explore the correlations between expression of miRNA-21 and PTEN and the invasion, metastasis of colorectal carcinoma.Methods The mRNA level of miRNA-21 and mRNA and protein levels of PTEN were assayed by real-time quantitative PCR and immunohistochemical methods respectively in 65 colorectal carcinoma specimen.Results The expression of miRNA-21 mRNA in cancer tissues was significantly higher than that in paracancerous tissues (t =3.50, P ＜ 0.05).Both mRNA and protein expressions of PTEN in tumor tissue were significantly lower than that in paracancerous tissues (t =7.35,t =12.23;P ＜ 0.05).Expression of miRNA-21 mRNA, PTEN mRNA and protein were obviously related with depth of invasion(F =18.36 ,F =17.26 ,F =12.83;P ＜ 0.05), Dukes stage (F =31.25, F =24.43, F =57.12;P ＜ 0.05) and lymph node metastasis (t =5.85, t =2.18, t =4.05;P ＜ 0.05), while it was not related to tumor differenciation and tumor location (F =7.39, t =4.62;F =7.78, t =1.29;F =5.14,t =1.37;P ＞ 0.05).Positive relation between miRNA-21 and PTEN in colorectal carcinoma was identified (r =-0.994, r =-0.927;P ＜ 0.05).Conclusion High expression of miRNA-21 and low expression of PTEN are both closely associated with invasion and metastasis of colorectal carcinoma.