A total of 152 strains of endophytic fungi were isolated from the barks of Eucommia ulmoides in three regions (Lueyang country, Zunyi country, Cili country). Based on morphological characteristics and analysis of ITS sequences, these strains were identified into 8 genera. Thereinto Phomopsis, Diaporthe and Alternaria were common genera to Eucommia barks from different sites. But the dominant genus was different: Alternaria was the dominant genus in the barks from Cili country, and Phomopsis was the dominant genus from Zunyi country, then Diaporthe was the one from Lueyang country. According to the similarity coefficient, the composition of the endophytic fungi was distinctly different between the barks from three sites. The diversity and species richness in Lueyang country and Cili country were found higher than those in Zunyi country. The evenness of endophytic fungi was 0.936 5 in Lueyang county, which was higher than 0.737 1 or 0.641 0 in Cili county or Zunyi county, respectively. After phylogenic analysis and calculating the genetic distances of typical strains belong to Phomopsis and its perfect stage--Diaporthe, there was very high genetic diversity in the two genera from our study. In conclusion, the community structure and diversity of endophytic fungi were significant different in Eucommia barks from the three habitats.
DNA, Fungal ; genetics ; DNA, Intergenic ; genetics ; Ecosystem ; Endophytes ; classification ; physiology ; Eucommiaceae ; microbiology ; Fungi ; classification ; genetics ; physiology ; Phylogeny ; Plant Bark ; microbiology

OBJECTIVETo explore the relative factors on the failure in digit replantation in order to take preventions to control the risk factors.

METHODSFrom January 2013 to December 2013, 236 consecutive patients (311 fingers) underwent digit replantation were collected to analyze retrospectively, involving 183 males and 53 females with an average age of 34.5 years old ranging from 2 to 62 years old (6 cases under 6 years old and 230 cases elder than 6 years old). There were 51 thumbs, 87 index fingers, 78 middle fingers, 63 ring fings and 32 little thumbs. Forty cases(forty fings) who were failured as the observation group, the others as the control group. The factors of age, gender, finger, cause of injury, smoking history, ischemia duration, plane of division, condition of venous drainage and condition of arterial repair we assessed.

RESULTSAll 236 cases with 311 fingers were replanted, 40 fingers were failured after operation. The relative factors on the failure in digit replantation included smoking history, cause of injury, plane of division, condition of venous drainage and condition of arterial repair (P< 0.05). There were no significant correlation between the failure and age, gender, finger and ischemia duration (P>0.05).

CONCLUSIONSmoking history, causes of injury, plane of division, condition of venous drainage and condition of arterial repair are risks of failure in digit replantation. Before choosing the type of operation, it should be think about the patient's general conditions, injury status, grasp firmly the operative indications and actively carry out surgical treatment.

Adolescent ; Adult ; Child ; Female ; Finger Injuries ; surgery ; Fingers ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Replantation ; Retrospective Studies ; Risk Factors ; Thumb ; injuries ; surgery ; Treatment Failure ; Young Adult

BACKGROUNDHeart failure (HF) is a major cause of morbidity and mortality worldwide, but current treatment modalities cannot reverse the underlying pathological state of the heart. Gene-based therapies are emerging as promising therapeutic modalities in HF patients. Our previous studies have shown that recombinant adeno-associated viral (rAAV) gene transfer of Sarco-endoplasmic reticulum calcium ATPase (SERCA2a) can be effective in treating rats with chronic heart failure (CHF). The aim of this study was to examine the effects of SERCA2a gene transfer in a large HF animal model.

METHODSHF was induced in beagles by rapid right ventricular pacing (230 beats/min) for 30 days. A reduced rate ventricular pacing (180 beats/min) was continued for another 30 days. The beagles were assigned to four groups: (a) control group (n = 4); (b) HF group (n = 4); (c) enhanced green fluorescent protein group (n = 4); and (d) SERCA2a group (n = 4). rAAV1-EGFP (1 x 10(12) microg) and rAAV1-SERCA2a (1 x 10(12) microg) were delivered intramyocardially. SERCA2a expression was assessed by Western blotting and immunohistochemistry.

RESULTSFollowing 30 days of SERCA2a gene transfer in HF beagles its protein expression was significantly higher than in the HF group than in the control group (P < 0.05). Heart function improved along with the increase in SERCA2a expression. Left ventricular systolic function significantly improved, including the ejection fraction, left ventricular systolic pressure, maximal rate of rise of left ventricular pressure (+dp/dt(max)), and the maximal rate of decline of left ventricular pressure (-dp/dt(max)) (P < 0.05). Left ventricular end-diastole pressure significantly decreased (P < 0.05). The expression of SERCA2a in the myocardial tissue was higher in the SERCA2a group than in the HF group (P < 0.05).

CONCLUSIONSIntramyocardial injection of rAAV1-SERCA2a can improve the cardiac function in beagles induced with HF. We expect further studies on SERCA2a's long-term safety, efficacy, dosage and the optimization before using it in humans with HF.

Animals ; Blotting, Western ; Disease Models, Animal ; Dogs ; Echocardiography ; Genetic Therapy ; methods ; Green Fluorescent Proteins ; genetics ; metabolism ; Heart ; physiology ; Heart Failure ; therapy ; Hemodynamics ; Immunohistochemistry ; Myocardium ; metabolism ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; genetics ; physiology

Objective Overexpression of SERCA2a could improve cardiac function in human and experimental heart failure(HF)models.We observed the proteomics changes post SERCA2a overexpression in a pacing induced HF model in dogs.Methods Beagles were divided into four groups:control group,HF group(230 beats/min for 4 weeks),HF+EGFP group(myocardial injection of 1 × 1012 v.g recombinant adeno-associated virus carrying enhanced green fluorescent protein gene,rAAV2/1-EGFP)and HF+ SERCA2a group ( myocardial injection of 1 × 1012 v.g recombinant adeno-associated virus carrying SERCA2a gene,rAAV2/1-SERCA2a).Thirty days after gene transduction,left ventficular systolic and diastolic functions were measured by echoeardiography and invasive hemodynamics in all animals.By use of 2-dimensional gel electrophoresis(2-DE),-500 distinct protein spots were detected in myocardium of all animals.Protein spots observed to be altered between failing and SERCA2a overexpressed hearts were subjected to tryptic peptide mass fingerprinting for identification by MALDI-TOF mass spectrometry in combination with LC/MS/MS analysis.Results At 30 day after gene transfer,HF signs were significantly reduced,cardiac function[LVSP:(214.72±31.74)mm Hg(1 mm Hg=0.133 kPa)vs.(139.32±36.79)mm Hg,+dp/dtmax:(6779.43±217.58)mm Hg/s vs.(2746.85±931.23)mm Hg/s and -dp/dtmax:(-4341.42±322.02)mm Hg/s vs.(r-2531.14 ±616.15)mm Hg/s,LVEDP:(21.86±6.95)mm Hg vs.(59.78±6.92)mm Hg]significantly improved in HF+SERCA2a dogs than those in HF+ EGFP group(all P＜0.05)and parameters were comparable between HF+SERCA2a and control groups.We identified alterations in the expression level of more than 10 proteins in myocardium.These protein changes were observed mainly in two subcellular compartments:the cardiac contractile apparatus and metabolism/energetics.Conclusion These results showed that overexpression of SERCA2a could improve cardiac function accompanied with numerous alterations in protein expressions involved in calcium handling,myofibrils,and energy production in this dog model of chronic heart failure.

OBJECTIVETo assess the association between 2 single nucleotide polymorphisms (SNPs) located in exonic regions of matrix metalloproteinase-10 (MMP-10) gene and instability of carotid plaques in a Han Chinese population.

METHODSFive hundred and eighty-five patients were divided into carotid vulnerable plaque group (n=206) and stable plaque group (n=379) based on results of carotid B-mode ultrasonography. The SNPs were genotyped by real-time polymerase chain reaction using an ABI 7300 TaqMan platform.

RESULTSThe distribution of rs17435959 between the two groups was significantly different at both genotypic (GC+CC vs. GG, P=0.006, OR=2.012) and allelic levels (C vs. G, P=0.001,OR=2.160). Above differences have remained significant with binary logistic regression analysis (P=0.007, OR=2.022; P=0.002, OR=2.104). The minor allele frequency of rs17293607 was 0.56%.

CONCLUSIONAbove findings suggested that rs17435959 of the MMP-10 gene is associated with carotid vulnerable plaque in ethnic Chinese Hans. The C allele may be a susceptible predictor for carotid vulnerable plaque.

Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Matrix Metalloproteinase 10 ; genetics ; Middle Aged ; Plaque, Atherosclerotic ; enzymology ; genetics ; Polymorphism, Single Nucleotide

Objective To observe the expression of extracellular matrix metalloproteinase inducer (EMMPRIN) in the unstable plaque of patients with acute coronary syndrome (ACS),and the impact of leukotriene B4 (LTB4) on the EMMPRIN expression in macrophages.Methods The EMMPRIN expression was detected by immunohistochemistry in 11 unstable plaques from patients with ACS.Protein expression of EMMPRIN was evaluated by Western blot on macrophages differentiated from THP-1 which were stimulated with LTB4 in the absence or presence of LTB4 antagonist U75302.There are 8 study groups:1-THP-1,2 -8-the macrophages derived from THP-1,2 - 6- macrophages were stimulated by LTB4 (0,10-10,10-9,10-8 and 10-7mol/L) for 24 h,7 -8-the macrophages were pretreated by 10-6 mol/L or 10-7 mol/L U75302 2 h before the LTB4 ( 10-7 mol/L) stimulation.Results Abundant EMMPRIN expression was detected in macrophages and smooth muscle cells of unstable plaques from ACS patients.As to the THP-1 derived macrophages,EMMPRIN expression was significantly upregulated in a concentration-dependent manner in LTB4 stimulated groups,which was significantly higher in group 3 -6 than in the THP-1 group ( group 1 ) and macrophages group ( group 2 ) ( all P ＜ 0.05 ) and pretreatment with U75302 significantly reduced the LTB4 induced upregulation of EMMPRIN in a dose-dependent manner (P ＜ 0.05).Conclusion EMMPRIN expression is enhanced in macrophages and smooth muscle cells on unstable coronary artery plaques from ACS patients.LTB4 could stimulate EMMPRIN expression on THP-1 derived macrophages suggesting that LTB4 and EMMPRIN might be both involved in the formation and progression of unstable plaques,future studies are warranted to explore if LTB4 and EMMPRIN antagonists are effective or not for treating patients with ACS.

Objective: To analyze the genetic landscape of 52 fusion genes in patients with de novo acute lymphoblastic leukemia (ALL) and to investigate the characteristics of other laboratory results. Methods: The fusion gene expression was retrospectively analyzed in the 1 994 patients with de novo ALL diagnosed from September 2016 to December 2020. In addition, their mutational, immunophenotypical and karyotypical profiles were investigated. Results: In the 1 994 patients with ALL, the median age was 12 years (from 15 days to 89 years). In the panel of targeted genes, 15 different types of fusion genes were detected in 884 patients (44.33%) and demonstrated a Power law distribution. The frequency of detectable fusion genes in B-cell ALL was significantly higher than that in T-cell ALL (48.48% vs 18.71%), and fusion genes were almost exclusively expressed in B-cell ALL or T-cell ALL. The number of fusion genes showed peaks at<1 year, 3-5 years and 35-44 years, respectively. More fusion genes were identified in children than in adults. MLL-FG was most frequently seen in infants and TEL-AML1 was most commonly seen in children, while BCR-ABL1 was dominant in adults. The majority of fusion gene mutations involved signaling pathway and the most frequent mutations were observed in NRAS and KRAS genes. The expression of early-stage B-cell antigens varied in B-cell ALL patients. The complex karyotypes were more common in BCR-ABL1 positive patients than others. Conclusion: The distribution of fusion genes in ALL patients differs by ages and cell lineages. It also corresponds to various gene mutations, immunophenotypes, and karyotypes.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Gene Expression ; Genes, ras ; Humans ; Infant ; Infant, Newborn ; Middle Aged ; Oncogene Fusion ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/metabolism* ; Retrospective Studies ; Young Adult