Effects of Shenfu yixin granule on mitochondrial autophagy of cardiomyocytes in rats with heart failure after acute yocardial infarction / 中国药房

OBJECTIVE To study the effects of Shenfu yixin granule on mitochondrial autophagy of cardiomyocytes in rats with heart failure after acute myocardial infarction. METHODS The model of heart failure after acute myocardial infarction was established by ligaturing the anterior descending branch of the left coronary artery in rats. The model rats were divided into model group，Shenfu yixin granule low-dose and high-dose groups （1.76，8.8 g/kg），Fosinopril sodium tablets group （positive control ，4 mg/kg），sham operation group was set up （only threading without ligation at the same position ），with 8 rats in each group. After 4 weeks of drug intervention ，the hemodynamic indexes of rats in each group were measured by physiological recorder. The pathological changes of myocardial tissue were observed in each group. The level of oxidative stress in cardiomyocytes ， mitochondrial membrane potential ，protein expression of PTEN-induced putative kinase 1（PINK1），E3 ubiquitin ligase Parkin and ubiquitin binding protein P 62 in myocardial tissue of rats in each group were detected. RESULTS Compared with sham operation group ，the pathological injuries such as myocardial fiber morphology disorder and inflammatory cell infiltration were serious. The left ventricular end systolic pressure （LVESP），maximum rate of rise of left ventricular internal pressure （+dp/dtmax）， maximun rate of decrease of left ventricular internal pressure （－dp/dtmax），total antioxidant capacity ，mitochondrial membrane potential，PINK1，Parkin and P 62 protein expression were significantly decreased in model group （P＜0.01）. The left ventricular end diastolic pressure （LVEDP），the level of reactive oxygen species and the activity of reduced nicotinamide adenine dinucleotide phosphate in left ventricular ischemic cardiomyocytes were significantly increased （P＜0.01）. Compared with model group ，the pathological injuries of myocardial tissue in intervention groups were alleviated ，and above indexes were improved in varying degrees（P＜0.01 or P＜0.05）. CONCLUSIONS Shenfu y ixin granule can reduce the level of oxidative stress and alleviate heart failure after acute myocardial infarction ，which may be related to the activation of Parkin-dependent pathway to strengthen mitochondrial autophagy and reduce mitochondrial dysfunction.

Identification of a COL2A1 mutation in a Chinese family with Stickler syndrome type 1 via whole exome sequencing / 中华实验眼科杂志

Objective:To identify the disease-causing mutation in a Chinese family with Stickler syndrome type 1.Methods:The pedigree investigation was conducted.A Chinese family with Stickler syndrome type 1 was enrolled in the Shantou International Eye Center in June 2012.Medical history collection and clinical examinations, such as vision, intraocular pressure, slit lamp microscopy and fundus, were carried out in all the included family members and the diagnosis was made by clinical experts.Total genomic DNAs were extracted from the peripheral blood samples (5 ml) obtained from 5 patients and 4 healthy members.The potential variant of the proband's father Ⅲ-5 were screened by whole exome sequencing (WES) and stepwise bioinformatic analysis.The segregation and mutation conformation of the variant was verified by Sanger sequencing.The pathogenicity of the variant was predicted by SIFT, Polyphen2, and MutationTaster.Conservation and three-dimensional structure of amino acid mutation were analyzed by multiple sequence alignment and UniProt.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Joint Shantou International Eye Center (No.EC20110310[2]-P02).Written informed consent was obtained from each subject or the guardian.Results:An autosomal dominant inherence in 39 members of 4 generations including 15 patients and 24 phenotypically normal members was found in the family.The proband (Ⅳ-4) showed high myopia, retinal detachment and strabismus in the right eye, and the left eye was blind.A patient (Ⅲ-5) showed high myopia and cataract in the right eye, atrophy in the left eye.A patient (Ⅳ-9) showed binocular high myopia.A heterozygous variation, c.1693C>T: p.Arg565Cys, within the exon 26 of COL2A1 gene was revealed in patient Ⅲ-5, which was only found in the patients and not in phenotypically normal members, indiacating co-separation in this family.The variant was predicted to be a severe damage by SIFT, Polyphen2 and MutationTaster.The amino acid mutation at position 565 was highly conservative among human, mouse, rat, bovine and Xenopus laevis, which caused the arginine to cysteine substitution at the X position in triple helix repeat region Gly-X-Y, affecting the function of fibrous protein and becoming pathogenic. Conclusions:Variant c.1693C>T: p.Arg565Cys in COL2A1 gene is disease-causing in this family and this is the first report about the variant in China.

Compound heterozygous mutations of CRB1 gene in a Chinese family with Leber congenital amaurosis by whole exome sequencing / 中华实验眼科杂志

Objective To investigate the disease-causing mutation in a family with Leber congenital amaurosis (LCA).Methods A Chinese Han pedigree with LCA from Chaoshan area was recruited in Shantou International Eye Center in August 2011.The clinical features of the families were evaluated,including medical history,best corrected visual acuity,intraocular pressure and fundus photography.The peripheral blood sample of 5 ml was collected from each of the family members for the extraction of genomic DNA.DNA of the proband was investigated by whole exome sequencing (WES) and was filtered for function of variants and inheritance pattern.Then,Sanger sequencing was performed to confirm the WES result on all the participating subjects in the pedigree.Results There were 11 families of 3 generations in this pedigree,and 2 female LCA patients were found (Ⅱ 2 and Ⅱ4) who were sisters.The parents (Ⅰ-1 and Ⅰ-2) and children (Ⅲ-1,Ⅲ-2,Ⅲ-3 and Ⅲ-4) of the patients showed normal phenotype,suggesting an autosomal recessive pattern.The patients appeared severe visual impairment during early childhood.Ophthalmic examination showed diffuse pigmentation on the retina and attenuation of retinal artery in both patients.WES of proband revealed two compound heterozygous mutations (c.2234C ＞T,p.T745M;c.3488G＞T,p.C1163F) of the CRB1 gene.Sanger sequencing confirmed the mutations in both patients (Ⅱ-2 and Ⅲ-4),and the parents of the patients were found to carry one mutations respectively and the other subjects with normal phenotype had neither none or only one mutation.Conclusions The compound heterozygous mutation of c.2234C＞ T,p.T745M and c.3488G＞T,p.C1163F in CRB1 is responsible for LCA pathogenesis this Chinese Han pedigree.

EP1 receptor mediates Adriamycin-induced podocyte injury through activating p38 MAPK / 中华肾脏病杂志

Objective To investigate the effect of prostaglandin E2 receptor 1 (EP1) on Adriamycin (ADR)-induced glomerular podocytes injury and its possible mechanism.Methods (1) In vivo experiments:6-8 weeks old male Balb/c mice were randomly divided into four groups:Control group;ADR group;EP1 agonist 17-phenyl PGE2+ADR group;EP1 antagonist SC-19220+ADR group.The mouse model of nephrotic syndrome was induced by injection of ADR (10 mg/kg) into tail vein,and then EP1 agonist (1 μg/g) and antagonist (25 μg/g) were administered respectively.Six weeks later,all mice were sacrificed and urine,blood and kidney tissues were collected.Detecting urine protein,blood chemistry,changes of renal pathology and podocyte-related proteins,electron microscopy changes of podocytes.(2) In vitro experiments:Podocytes were cultured in vitro and divided into different groups:Control group;ADR group (0.2 μmol/L);EP1 agonist (0.1,1,10 μmol/L)+ADR (0.2 μmol/L) group;antagonist (0.1,0.5,1 μmol/L)+ ADR (0.2 μmol/L) group.The proliferation of podocytes was measured by CCK-8.Expression of PGE2 in podocytes was detected by ELISA.Indirect immunofluorescence was used to determine the localization of podocyte-related proteins nephrin,podocin and CD2AP.Expression of nephrin,podocin,CD2AP,COX2 in podocytes was detected by Western blotting and Real-time quantitative PCR.p38 MAPK or phospho-p38 MAPK was measured by Western blotting as well.Flow cytometry was used to detect cell apoptosis.Results (1) In vivo experiments:Compared with control group,obvious proteinuria,blood biochemical changes and renal pathological changes were observed in ADR group,proteinuria,blood biochemical and renal pathological changes were more serious in mice dealt with agonist,while antagonist could reduce ADR-induced injury (all P ＜ 0.05).Results of immunohistochemistry showed that the expression of podocyterelated proteins nephrin,podocin and CD2AP in ADR group were significantly lower than those in control group,and EP1 agonist could further inhibit expression of these proteins,while antagonist could reverse this inhibitory effects (P ＜ 0.05).Electron microscopic results showed that mice in ADR group appeared foot enlargement and fusion,and the agonist group further aggravated the injury,while antagonist intervention could inhibit the injury of podocytes.(2) In vitro experiments:Compared with control group,expression of PGE2 and COX2 were increased;mRNA and protein expression of nephrin,podocin,CD2AP were decreased,p38 MAPK activity and podocytes apoptosis were increased in ADR group (P ＜ 0.05);Agonist could aggravate podocytes damage (P ＜ 0.05),while Antagonist could downregulate the expression of PGE2 and COX2,promote the expression of nephrin,podocin and CD2AP,and inhibit the activity of p38 MAPK and podocytes apoptosis (P ＜ 0.05).The addition of p38 MAPK inhibitor(10 μmol/L) could reduce the inhibitory effect of EP1 agonist on the expression of podocyterelated proteins nephrin,podocin and CD2AP (P ＜ 0.05).Conclusions EP1 receptor may activate the p38 MAPK signaling pathway to inhibit podocytes-related proteins nephrin,podocin and CD2AP,as well as mediate the ADR induced podocyte injury.Inhibition of EP1 receptor however have a protective effect.

Practice research on rehabilitation effect of using interactive graphics in humanistic care in lung cancer patients underwent thoracic surgery / 中国实用护理杂志

Objective To explore rehabilitation effect of graphic interactive education in humanistic care in lung cancer patients underwent thoracic surgery. Methods Two groups were randomly formed based on a total of 95 lung cancer patients underwent open thoracic surgery from January 2016 to December 2016: normal group (47 people) and experimental group (48 people). The patients in normal group were treated with routine health method. The experimental group was cared with graphic interactive education, i.e. cares were applied based on humanistic thoughts. Results The average rehabilitation accuracy score of the patients in the experimental group (18.833 ± 2.999) was significantly higher than the one in the normal group (12.255 ± 3.247). The difference is statistical significant (t=10.261, P < 0.01). In the experimental group the patients were more satisfied and the compliance of upper limb during rehabilitation exercise was significantly higher. The difference comparing to the normal group was statistically significant (Z=-3.419,-6.232, P<0.01). Conclusions The rehabilitation effect of using interactive graphics in humanistic care is apparent. This method is recommended to be used as a highlight of special high qualified nursing service to improve patients′satisfaction.

Extraction Technology of Flavone Ingredients of Herba Epimedii by Multi-index Uniform Design / 医药导报

Objective To optimize extraction technology of flavonesingredients from Herba Epimedii by uniform design. Methods Ultraviolet spectrophotometry was adopted to determine the content of total flavonoids. Content of epimedin A, epimedin B,epimedin C,icariin and baohuosideⅠ were determined by HPLC. U10?( 108 ) uniform design was used to conduct the multiple comprehensive evaluation of six ingredients and comprehensively analyze the influence of the concentration and amount of ethanol, extracting time on extraction of flavonesin gredients from Herba Epimedii. Results The uniform design experiment showed that 15-fold weight of 60% ethanol,extracting 2 times and each time with 165 min were the optimum extraction condition. Conclusion The method is easy and reasonable to handle,has stable and reliable results,and good repeatability and feasibility.It can be applied in industrial production.