Objective To lay the foundation for analyzing the mechanism of liver cell injury caused by mtDNA deletion and mutation in patients with obstructive jaundice. Methods 30 patients were randomly selected as obstructive jaundice group (case group) and 10 patients as control group according to the strict condition. Author makes use of the methods of PCR amplification of the entire human mitochondrial genome in 17 mismatch-specific overlapping fragments and gene sequencing results to Preliminary estimate the localizathion of hepatocyte mtDNA damage in patients with obstructive jaundice. Result Deletions and length of partial liver cells were 8429-9591 of about 1. 1 kb, 16024-60 of about 0. 6 kb, 1889-3031 of about 1. 1 kb and 4977bps common deletion and the high mutation rate of some bases in D-loop region. Conclusion There are multiple mtDNA deletions and multiple point mutations in patients with obstructive jaundice

ObjectiveTo investigate the mechanism of modified Shenhong Tongluo prescription on cell apoptosis in rats with myocardial ischemia-reperfusion injury （MIRI）. MethodSixty Sprague-Dawley （SD） rats were randomly divided into a blank group， a model group， low-， medium-， and high-dose groups of modified Shenhong Tongluo prescription， and a simvastatin group. Except for the blank group， a rat model of MIRI was prepared by ligating the left anterior descending coronary artery. Starting from the first day after successful modeling， the blank group （1.0 mL·kg^-1 physiological saline）， model group （1.0 mL·kg^-1 physiological saline）， low-， medium-， and high-dose groups of modified Shenhong Tongluo prescription （1.031， 2.063， and 4.126 g·kg^-1 Shenhong Tongluo prescriptiona standard concentrate）， and simvastatin group （0.71 mg·kg^-1 simvastatin） were orally administered once daily for 2 weeks. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of cardiomyocytes. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of serum creatine kinase isoenzyme （CK-MB）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α）. TdT-mediated dUTP nick-end labeling（TUNEL） staining was used to detect the apoptosis rate of rat cardiomyocytes. Western blot was used to detect the expression levels of apoptosis-related proteins B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and caspase-3. ResultCompared with the blank group， in the model group， HE staining showed disturbed arrangement of cardiomyocytes， incomplete fibers， focal necrosis of cardiomyocytes， and inflammatory cell infiltration； serum CK-MB， IL-6， and TNF-α levels were significantly increased （P<0.05）； apoptosis rate of cardiomyocytes was significantly increased （P<0.01）， with significantly increased expression levels of Bax and Caspase-3 proteins， and significantly decreased Bcl-2 expression （P<0.05）. Compared with the model group， the low-， medium-， and high-dose groups of modified Shenhong Tongluo prescription significantly reduced CK-MB， IL-6， and TNF-α levels （P<0.05）， significantly downregulated cardiomyocyte apoptosis rate （P<0.05）， significantly decreased Bax and Caspase-3 proteins， and significantly increased Bcl-2 expression levels （P<0.01）. In the modified Shenhong Tongluo prescription groups， the expression levels of Bax and Caspase-3 proteins significantly decreased with increasing dosage， while the expression level of Bcl-2 significantly increased with increasing dosage of modified Shenhong Tongluo prescription （P<0.05）. ConclusionShenhong Tongluo prescription can alleviate myocardial tissue pathological damage and reduce myocardial cell apoptosis， possibly by inhibiting Caspase-3 and Bax expression and promoting Bcl-2 expression.

AIM: To establish the drug release method of Compound Salvia Multi-component released Doublelayer Tablet and investigate its in vitro drug release behavior and the influencing factors of the drug release rate.METHODS: The method of evaluating in vitro release rate of double-layer tablet was established with salvianolic acid B and gindenoside Rg_1 as index,The influencing factors of the release of rapid release layer and sustained release layer of double-layer tablet and their in vitro drug release behavior were studied.RESULTS: The rapid release layer showed the quick releasing effect; The drug release curve of sustained release layer accorded with Ritger-Peppas equation; artificial gastric juice had significant effect on the release curve of gindenoside Rg_1 which in double-layer tablets,but had no significant effect on the release curve of salvianolic acid B.CONCLUSION: The evaluation of in vitro release rate of Compound Salvia Multi-component released Double-layer Tablet shows good properties of fast and sustained release and clinic application is achieved.

Objective To evaluate the relationship between echocardiographic epicardial adipose tissue thickness(EAT) and the presence and severity of coronary artery disease(CAD). Methods One hundredand forty-seven patients (101 patients with CAD and 46 patients with normal coronary arteries by diagnostic coronary angiography) were enrolled. EAT thickness was measured using 2-D echocardiographic parasternal long-and short-axis views. EAT thickness measurements were compared with angiographic findings. Results EAT was significantly higher in CAD group comparison to control group [(7.41 ± 1.63)mm vs (4.41±1.60) mm, P ＜0.01 ]. Furthermore, EAT increased with the severity of CAD [(8.53 ± 1.00)mm vs (6.36 ±1.73)mm, P ＜0.01]. Gensini's score significantly correlated with EAT (r = 0.71, P ＜0.01 ). EAT thickness ≥5.35 mm had 87.13% sensitivity and 80.42% specificity (ROC area 0. 89, P = 0.01,95% CI [0.84 - 0.9;]) for predicting CAD. Conclusions EAT thickness, which is easily and non-invasively evaluated by transthoracic echocardiography, can be an adjunctive marker to classical risk factors for the prediction of CAD, it was significantly correlated with the severity of coronary artery disease.

Objective To investigate the correlation between epicardial fat thickness and non dipper hypertension.Methods A total of 150 subjects was included in the study,of which 50 were in the non dipper hypertension group,the same in the non dipper hypertension group and the healthy control group.History collection and routine laboratory tests,ultrasonic measurement of epicardial fat thickness,and 24 hour ambulatory blood pressure monitoring were carried on all subjects.Epicardial fat thickness between groups was compared to primarily analyze the correlation of epicardial fat thickness and non dipper type hypertension.The optimal screening positive value in epicardial fat thickness of non dipper type primary hypertension was obtained by receiver operating characteristic (ROC) curve and maximum Youden index.Results When non dipper hypertension group and non-dipper hypertension group were compared,epicardial fat thickness was significantly increased [(6.30 ± 0.94) mm vs (5.92 ± 0.75) mm,P ＜ 0.05],as compared dipper hypertension group to healthy group,the epicardial fat thickness was significantly increased [(5.92 ±0.75)mm vs (5.50 ±0.13)mm,P ＜0.05].Epicardial fat thickness and non dipper type primary hypertension were linearly related (r =0.43,P ＜ 0.05),and epicardial fat thickness in diagnosis of non dippers primary hypertension optimal screening positive value was 6.01 mm.Conclusions There is a close relationship of epicardial fat thickness and non dipper hypertension.

Objective To analyze the effectiveness evaluation of cardiovascular drugs which have been developed on the CYP2C9 target protein by multi-layer fuzzy evaluation technology .Methods The multi-layer fuzzy evaluation method was used to evaluate the effectiveness of cardiovascular drugs interacting with the CYP 2C9 protein and to construct the index system that affects drug efficacy .Results and Conclusion The index system was used to study such cardiovascular drugs as valsartan and to score the drug effectiveness of individual samples .The results were consistent with actual drug treatment and were well confirmed .The results contribute to evaluation of personalized medication .

OBJECTIVETo study the genome sequence of hepatitis A virus L-A-1 strain which has been applied for live attenuated vaccine production in China, to compare with other HAV strains, to understand some characteristics of L-A-1 strain, and to find the mechanism of attenuation and cell adaptation.

METHODSGenome fragments were prepared by antigen-capture PCR from infected cell (2BS), PCR products were cloned into T vector, sequenced and analyzed by using bioinformatics program.

RESULTSAnalysis of the genomic sequences(nt 25-7,418) showed that the open reading frame contains 6,675 nucleotides in length encoding 2,225 amino acids. Sequence homology comparison showed 98.00% and 94.00% homology at nucleotide level, and 98.51% and 98.65% homology at amino acid level with international strains MBB and HM 175, respectively. Through comparison with other attenuated, cell adapted and cytopathic effect (CPE) strains, L-A-1 strain had mutation at nt 152, 591, 646, 687 and insertion at nt 180-181 in 5?NTR and had mutation at nt 3,889 (aa 1 052-Val) in 2B region, these mutations and insertion are molecular basis for cell adaptation; mutation at nt 4,185 (aa 1 152-Lys) in 2C region should be attenuated marker; deletion in 3A region (nt 5,020-5,025) that caused two amino acids deletion is virus fast growth basis.

CONCLUSIONThrough analyzing L-A-1 strain genomic sequence, certain sites related to cell adaptation and attenuation were found.

Adaptation, Biological ; genetics ; Amino Acid Sequence ; Base Sequence ; Gene Deletion ; Genome, Viral ; Hepatitis A Vaccines ; genetics ; Hepatitis A virus ; genetics ; growth & development ; Mutation ; Open Reading Frames ; genetics ; Sequence Homology ; Vaccines, Attenuated ; genetics

Aim To investigate the sites and mechanisms of action of Ginseng-Rhodiola rosea in the treat ment of myocardial ischemia-reperfusion injury ( MI-RI) via using network pharmacology approach, molecu¬lar docking techniques and experimental studies. Methods The active ingredients and targets of Gin¬seng-Rhodiola rosea were screened through the TCMSP database and literature supplementation, and the GEN-EC ARDS ,DISGENET and DRUGBANK databases were searched to obtain the targets of MIRI. Functional pro¬tein interaction networks (PPIs) and the STRING database were used to screen out core targets. The DAVID database was also selected for gene ontology functional analysis ( GO) and KEGG signaling pathway enrich¬ment analysis. Lastly, the preliminary validation was performed with the help of molecular docking techniques and experimental studies. Results Forty-three active ingredients and 348 potential targets of Ginseng-Rhodiola were obtained, and targets such as IL-6 , TNF-α and VEGFA were found to be closely related to MIRI, mainly involving TNF, PDK-Akt, HIF-1 and other signaling pathways.The molecular docking results showed that soysterol, ginsenoside rh2 and rhodioloside had good binding effects and high matching with IL-6, TNF-α,Caspase-3,VEGFA,MAPK1 and other targets, among which the best binding was between Caspase-3 and ginsenoside rh2. The results of the experimental study further showed that Ginseng-Rhodiola rosea could improve myocardial tissue necrosis after myocardial ischemia-reperfusion , reduce myocardial cell edema and vascular congestion, and decrease the expression levels of TNF-α and IL-6 in MIRI rats. Conclusions Ginseng-Rhodiola may modulate multiple targets such as IL-6,TNF-α, Caspase-3, VEGFA and MAPK1 through dousterol, ginsenoside rh2 and rhodiol glycosides to inhibit inflammatory response and oxidative stress, reduce cardiomyocyte damage and exert therapeutic effects on MIRI.

Objective　To clone full length of Kif1a gene promoter,and to construct and identify Kif1a gene promoter pGL3-Kif1a vectors.Methods　Recombinant of pGEM-T easy vector (2565 bp) including Kif1a gene promoter full sequence was used as a template in the polymerase chain reaction (PCR) to amplify its promoter sequence (1853 bp).The PCR product was directly cloned into the luciferase reporter vector pGL3-Basic.Then the products were identified by DNA sequencing,nest PCR and restriction enzyme digestion.Then it was transfected into SCG cells,and the luciferase activity of the cells was analyzed after transfection.Results　Restriction enzyme digestion and DNA sequencing confirmed that the sequenced segment (1853 bp) in the recombinant was identical to that in GenBank and the segment was inserted in right direction.The activity of pGL3-Kif1a was significantly higher than that in pGL3-basic vectors.Conclusion　The luciferase expression vector-pGL3-Kif1a containing Kif1a gene promoter full length sequence is constructed successfully.The construction of pGL3-Kif1a recombinant plasmid lay a foundation of the analysis of promoter activities,gene expression regulatory mechanism and signal transduction of Kif1a.

Humans ; Joint Dislocations ; Lunate Bone ; Wrist Injuries