Advances on research of structure and function of puromycin sensitive aminopeptidase / 药学学报

Puromycin-sensitive aminopeptidase (PSAP) belongs to the M1 family of aminopeptidases, characterized by the N-terminal substrate binding sequence GAMEN, the enzyme activity center HEXXH(X)18E motif, and the C-terminal ERAP-1-like superfamily structural domain. Encoded by the gene NPEPPS located at 17q21.32, PSAP consists of 919 amino acids and is widely distributed throughout the human body, with the highest expression in the brain, followed by the heart and skeletal muscle. It is also found in the liver, renal tubular epithelium, small intestine, large intestine epithelium, and gastric epithelial cells. PSAP primarily relies on its aminopeptidase hydrolytic activity to remove toxic protein aggregates such as Tau, poly Q, and Cu, Zn-superoxide dismutase 1, making it an important factor in the development of diseases such as Alzheimer's disease, Huntington's chorea, and tumors. Existing PSAP inhibitors include bestatin, amastatin, leuhistin, actinonin, and purinomycin, some of which are already available or in clinical trials. This review provides an overview of the structural and biological functions of M1 family aminopeptidases, with a focus on PSAP, to facilitate further research and targeted drug development.

Evaluation of left ventricular structural and functional abnormalities and vascular calcification in kidney transplant recipients and the effect of bone metabolism on them / 中华器官移植杂志

Objective:To evaluate left ventricular structural and functional abnormalities and vascular calcification in kidney transplant (KT) recipients, explore their influencing factors and examine the effects of mineral and bone disorders.Methods:From January 2017 to December 2019, retrospective analysis was performed for 292 KT recipients. Biochemical markers of bone metabolism, bone mineral density (BMD), left ventricular hypertrophy (LVH), left ventricular ejection fraction (LVEF), left ventricular diastolic function, coronary artery calcification (CAC) score and thoracic aortic calcification (TAC) score were assessed. Linear regression and binary Logistic regression analyses were employed for evaluating the influencing factors of cardiovascular parameters and the influence of abnormal mineral and bone metabolism.Results:Postoperative abnormalities in mineral and bone disorders were manifested mostly as hypercalcemia (8.9%, 26/292), hypophosphatemia (27.1%, 79/292), low 25-hydroxyvitamin D (25(OH)vitD) (67.0%, 196/292), hyperparathyroidismhigh parathyroid hormone (PTH) (50.6%, 148/292), elevated bone turnover markers and bone loss rate of 25%-30%. The prevalence of LVH, LVEF<50%, left ventricular diastolic dysfunction, high CAC score and high TAC score were 39.9%(116/292), 0%, 13.1%(38/292), 17.3%(50/292) and 39.9%(116/292) respectively. The results of multivariate analysis indicated that LVH was correlated positively with hypertension and serum calcium (Ca) (95% CI: 1.242-28.080, P=0.026; 95% CI: 1.714-277.584, P=0.018); LVEF was correlated positively with lumbar vertebrae BMD (95% CI: 0.000 1-0.005 5, P=0.041); Left ventricular diastolic dysfunction was correlated positively with age, diabetes and parathyroid hyperplasia/nodules (95% CI: 1.050-1.176, P<0.001; 95% CI: 2.118-43.813, P=0.003 and 95% CI: 1.419-9.103, P=0.007); High CAC score was correlated positively with recipient age and dialysis time (95% CI: 1.036-1.160, P=0.001; 95% CI: 1.009-1.041, P=0.002); High TAC score was correlated positively with age (95% CI: 1.095-1.215, P<0.001). Correlation analysis indicated that TAC was correlated positively with serum Ca ( r=0.233, P=0.003), bone-specific alkaline phosphatase (BALP)( r=0.325, P<0.001) and type Ⅰ collagen cross-linked N-terminal peptide (NTX)( r=0.204, P=0.011) and negatively with femoral neck BMD ( r=0.194, P=0.017). Conclusions:There is a high prevalence of left ventricular structural and functional abnormalities and vascular calcification. It is closely correlated with mineral and bone disorders.

B lymphocyte deficiency affects the structure of mouse heart and the composition of cardiac immune cells / 西安交通大学学报(医学版)

【Objective】 To explore the effect of B lymphocytes on cardiac structure and function and myocardial immune cells during heart development. 【Methods】 Echocardiography, immunofluorescence staining and flow cytometry were used to evaluate the composition of immune cells of the heart and the cardiac structure and function in wild-type (WT) mice and B-lymphocyte-deficient (μMT) mice, respectively. 【Results】 Compared with those of μMT mice, the ratio of heart weight to mouse weight (P<0.05), left ventricular mass (P<0.05) and the cross-sectional area of myocardial cells WT mice were significantly increased, while the ventricular ejection fraction was significantly decreased (P<0.05). The results of mRNA sequencing showed that WT mice and μMT mice differentially expressed genes were mainly enriched in the signal pathway of heart development and hypertrophic cardiomyopathy. The results of flow cytometry showed that WT mice had more Ly6g+ neutrophils, CD4+ positive T cells (P<0.001) and CD8+T cells (P<0.05) compared with μMT mice. 【Conclusion】 B-lymphocyte depletion alters the composition of cardiomyocyte immune cells, reduces left ventricular mass, and increases myocardial contractility.

Cardiac Structural and Functional Features in Patients With Type 2 Diabetes Mellitus and Heart Failure With Preserved Ejection Fraction:A Study Based on Propensity Score Matching / 中国医学科学院学报

Objective To investigate the cardiac structural and functional characteristics in the patients with heart failure with preserved ejection fraction (HFpEF) and type 2 diabetes mellitus (T2DM),and predict the factors influencing the characteristics. Methods A total of 783 HFpEF patients diagnosed in the Department of Geriatric Cardiology,the First Hospital of Lanzhou University from April 2009 to December 2020 were enrolled in this study.Echocardiography and tissue Doppler technique were employed to evaluate cardiac structure and function.According to the occurrence of T2DM,the patients were assigned into a HFpEF+T2DM group (n=332) and a HFpEF group (n=451).Propensity score matching (PSM)(in a 1∶1 ratio) was adopted to minimize confounding effect.According to urinary albumin excretion rate (UAER),the HFpEF+T2DM group was further divided into three subgroups with UAER<20 μg/min,of 20-200 μg/min,and>200 μg/min,respectively.The comorbidities,symptoms and signs,and cardiac structure and function were compared among the groups to clarify the features of diabetes related HFpEF.Multivariate linear regression was conducted to probe the relationship of systolic blood pressure,blood glucose,glycosylated hemoglobin,and UARE with cardiac structural and functional impairment. Results The HFpEF+T2DM group had higher prevalence of hypertension (P=0.001) and coronary heart disease (P=0.036),younger age (P=0.020),and larger body mass index (P=0.005) than the HFpEF group,with the median diabetic course of 10 (3,17) years.After PSM,the prevalence of hypertension and coronary heart disease,body mass index,and age had no significant differences between the two groups(all P>0.05).In addition,the HFpEF+T2DM group had higher interventricular septal thickness (P=0.015),left ventricular posterior wall thickness (P=0.040),and left ventricular mass (P=0.012) and lower early diastole velocity of mitral annular septum (P=0.030) and lateral wall (P=0.011) than the HFpEF group.Compared with the HFpEF group,the HFpEF+T2DM group showed increased ratio of early diastolic mitral filling velocity to early diastolic mitral annular velocity (E/e') (P=0.036).Glycosylated hemoglobin was correlated with left ventricular mass (P=0.011),and the natural logarithm of UAER with interventricular septal thickness (P=0.004),left ventricular posterior wall thickness (P=0.006),left ventricular mass (P<0.001),and E/e' ratio (P=0.049). Conclusion The patients with both T2DM and HFpEF have thicker left ventricular wall,larger left ventricular mass,more advanced left ventricular remodeling,severer impaired left ventricular diastolic function,and higher left ventricular filling pressure than the HFpEF patients without T2DM.Elevated blood glucose and diabetic microvascular diseases might play a role in the development of the detrimental structural and functional changes of the heart.

Relationship between preoperative glycosylated hemoglobin level and changes in structure and function of macular area in type 2 diabetes mellitus patients after phacoemulsification / 中国医师进修杂志

Objective:To explore the relationship between preoperative level of glycosylated hemoglobin (HbA 1c) and changes in structure and function of macular area in patients with type 2 diabetes mellitus (T2DM) after phacoemulsification. Methods:A total of 118 patients with T2DM and monocular cataract undergoing phacoemulsification in Hefei First People′s Hospital, were enrolled between January 2018 and June 2021. According to preoperative HbA 1c level, they were divided into high HbA 1c group (HbA 1c >7.0%, 55 cases) and normal HbA 1c group (HbA 1c ≤7.0%, 63 cases). The central macular thickness (CST), central volume (CV), subfoveal choroidal thickness (SFCT) and incidence of postoperative cystoid macular edema (PCME) between the two groups were compared. The risk factors of PCME were analyzed. Results:At 1 week, 1 month and 3 months after phacoemulsification, CST level in high HbA 1c group was significantly higher than that in normal HbA 1c group: (256.21±19.27) μm vs. (242.33 ± 14.63) μm, (277.41 ± 16.39) μm vs. (248.12 ± 11.54) μm, (242.51 ± 11.52) μm vs. (234.87 ± 9.34) μm, and there was statistical difference ( P<0.05). At 3 months after phacoemulsification, CV level in high HbA 1c group was significantly higher than that in normal HbA 1c group: (10.40 ± 0.51) mm 2 vs. (10.17 ± 0.47) mm 2 ( P<0.05). The incidence of PCME in high HbA 1c group was significantly higher than that in normal HbA 1c group: 25.45% (14/55) vs. 7.94% (5/63) ( P<0.05). Preoperative HbA 1c>7.0% and retinopathy were independent risk factors of PCME ( P<0.05). Conclusions:The preoperative HbA 1c level will affect retina and choroid structures in macular area of patients with T2DM and cataract after phacoemulsification. The risk of PCME is higher in patients with preoperative HbA 1c>7.0%.

Bioinformatics analysis on structure and function and expression vector construction of SARS-CoV-2 related protein TMPRSS2 / 中国药理学通报

Aim Human TMPRSS2 is a transmembrane serine protease.In this paper, the structure and func¬tion of the protein were systematically analyzed by bioinformatics, the codon was optimized and the pro- karvotie expression vector was constructed to explore the molecular mechanism of SARS-CoV-2 infecting host cells.Methods The recombinant expression vector pET-22b-TMPRSS2 was generated by molecular clo¬ning technology.The homology, functional sites, sub¬cellular localization, three-dimensional structure and evolutionary characteristics of TMPRSS2 protein were systematically analyzed by using analytical tools such as Protparam, NetPhos3.1, Blast, Clustal X2 and MEGA7.0.Results The prokarvotic expression plas- mid was constructed correctly; TMPRSS2 belongs to medium molecular weight protein, which is composed of 492 amino acid residues.The theoretical isoelectric point is 8.12, the molecular extinction coefficient is 118 145 L • mol~1 • cm"1 , and the half-life is 30 h; TMPRSS2 has 15 potential glycosylation sites and 49 possible phosphorylation sites.It is a transmembrane hydrophilie protein without signal sequenee.In addi¬tion, the protein has 13 potential B-cell epitopes and 7 T-eell epitopes.Seeondarv structure analysis showed that random coil accounted for the highest proportion of TMPRSS2 protein ( 0.453 3) , followed by extended strand (0.252 0).Sequence comparison and evolu¬tionary analysis showed that the highest sequence con¬sistency and closest genetic relationship with human TMPRSS2 was Pan troglodytes, followed by gorilla.Conclusions Human-derived TMPRSS2 protein is ev- olutionarilv conserved and functionally important.Hie results of this study can help to reveal the structure and mechanism of action of TMPRSS2 protein, provide ide¬as for the diagnosis and treatment of COYID-19, and accelerate the research and development process of new drugs targeting TMPRSS2 protein.

Effect of Baihe Dihuang Decoction on synaptic plasticity of hippocampus in rats with anxious depression / 中国中药杂志

To explore the effect of Baihe Dihuang Decoction on the synaptic plasticity of hippocampal neurons in rats with anxious depression. Fifty SD rats were randomly divided into normal group, model group, venlafaxine group(6.75 mg·kg~(-1)), high-dose Baihe Dihuang Decoction group(8.64 g·kg~(-1)) and low-dose Baihe Dihuang Decoction group(4.32 g·kg~(-1)). Chronic restraint stress(6 h) combined with corticosterone(ih, 30 mg·kg~(-1)) was used to establish an anxious depression model, and 7 days after modeling, the administration started and continued for 21 days. The anxiety and depression-like behaviors of the rats were evaluated. Golgi-Cox staining and electron microscopy were used to observe the morphology and ultrastructural changes of synaptic dendrites. Immunofluorescence was used to detect the expression of hippocampal synaptic plasticity protein synapsin-1 and postsynaptic density protein 95(PSD-95). Western blot method was used to detect the expression of functional protein synaptophysin(SYP) and synaptic Ras GTPase activating protein(SynGap). The results showed that the rats in the model group had obvious anxiety and depression-like behaviors, the hip-pocampal dendritic spine density and branch length were reduced, the number of synapses was cut, and the internal structure was da-maged. The average fluorescence intensity of synapsin-1 and PSD-95 was significantly reduced and the expression of SYP and SynGap also decreased. High-dose Baihe Dihuang Decoction could significantly improve the anxiety and depression-like behaviors of model rats, relieve synaptic damage, and increase the expression of synapsin-1, PSD-95, SYP, and SynGap proteins. Therefore, we believe that Baihe Dihuang Decoction can improve anxiety and depression behaviors by regulating the synaptic plasticity of hippocampal neurons.

Effects of salidroside on structure and function of erythrocyte membrane in high altitude polycythemia rats / 中草药

Objective: To investigate the effects of salidroside on the structure and function of erythrocyte membrane in rats with high altitude polycythemia (HAPC), and to provide the scientific basis for the mechanism of salidroside preventing and treating HAPC. Methods: A total of 40 Wistar rats were randomly divided into five groups, control group, HAPC model group, salidroside high-dose (200 mg/kg), medium-dose (100 mg/kg), and low-dose (50 mg/kg) groups, four female and four male rats in each group. In addition to the control group, the remaining four groups established the HAPC model. The rats in the control group and model group were ig administered with saline, and rats in salidroside group were treated with different doses of salidroside at the same time. The dosage volume was 10 mL/kg, and once a day for 40 d. After the administration, blood was collected from femoral artery of the rats. Biochemical and enzyme-linked immunoassay were used to determine the lipid fluidity of erythrocyte membrane, total cholesterol content, total phospholipids content, the content of phospholipid components including phosphatidic acid (PA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), the activity of Na+, K+-ATPase and Ca2+, Mg2+-ATPase on erythrocyte membrane, and the concentrations of Na+ and Ca2+ in erythrocyte. Results: Salidroside can significantly improve the lipid fluidity, total phospholipids, PA, PC, PE, and PS content of erythrocyte membrane, and improve the activity of Na+, K+-ATPase and Ca2+, Mg2+-ATPase of erythrocyte membrane in HAPC rats. Total cholesterol content on erythrocyte membrane, and the concentrations of sodium and calcium in erythrocyte were significantly reduced. Conclusion: These findings suggested that salidroside can improve the function of erythrocyte membrane and cell metabolic activities by regulating the lipid composition of erythrocyte membrane, thereby alleviating the symptoms associated with high altitude polycythemia.

Research progress in evaluating velopharyngeal structures and functions by magnetic resonance imaging / 口腔疾病防治

@#Normal development of the velopharyngeal structures is key to obtaining good velopharyngeal closure. In the assessment of velopharyngeal closure and normal pronunciation, a variety of instruments can be used to detect and assist in the diagnosis of velopharyngeal dysfunction. In the past, the assessment of velopharyngeal closure often used two-dimensional imaging or relied solely on the subjective assessment of the phonetician. With the development of science and technology, magnetic resonance imaging (MRI) has become widely used in the evaluation of velopharyngeal structures and functions as an ideal examination method. This article reviews the current capabilities and limitations in evaluating velopharyngeal closure, as well as recent research on the structures and functions of the velopharyngeal using static MRI, dynamic MRI, three-dimensional MRI reconstructions and diffusion tensor imaging (DTI) techniques; in addition, this work explores the role and significance of MRI technology in evaluating the structures and functions of the velopharyngeal. A review of the literature shows that static MRI is simple in terms of the scanning mode, has easily adjustable parameters, and clearly shows the anatomical structures of palatopharyngeal in resting or transient vocal states. Dynamic MRI can capture the anatomical changes of the palatopharyngeal in a more complex pronunciation state and obtain accurate dynamic images of the velopharyngeal closure process for the study of speech pathology. Three-dimensional MRI reconstructions are usually used in fine scanning of the velopharyngeal structures in a resting state; although this method takes a long time, the images obtained are clear and reliable. This approach can be used for three-dimensional reconstruction analysis and three-dimensional finite element analysis, and it can be used to help plan an operation and evaluate the effect of the surgery. DTI is a new method for observing the contractile function of muscles by observing the locus of water molecules in muscles. DTI can be used to analyze and study many muscles involved in velopharyngeal closure.

Effect of Tai Chi on Cardiac and Static Pulmonary Function in Older Community-Dwelling Adults at Risk of Ischemic Stroke: A Randomized Controlled Trial / 中国结合医学杂志

OBJECTIVE@#To evaluate the effect of tai chi exercise on cardiac and static lung function for older community-dwelling adults at risk of ischemic stroke.@*METHODS@#A total of 170 older community-dwelling adults (aged 55-75 years old) at risk of ischemic stroke were allocated to either tai chi training group (85 cases, five 60-min sessions of tai chi training per week for 12 weeks) or control group (85 cases, usual pbysical activity for 12 weeks) using a computer-generated randomization. The echocardiographic parameters of cardiac structure, cardiac function and static lung function were measured at baseline, after 12 weeks of intervention and additional 12-week follow-up period by a blinded professional staffmember using a color Doppler ultrasound imaging device or a cardiopulmonary function instrument. The t test and linear mixed model based on the intentionto-treat analysis principle was used to calculate the effect. The adverse effect was observed.@*RESULTS@#Most of echocardiographic parameters on the cardiac structure, cardiac function and static lung function between the tai chi group and control group did not have a significant difference either post 12-week intervention or additional 12-week follow-up period. Only three parameters involving in right ventricular diameter (P=0.024), main pulmonary artery diameter (P=0.002) and vital capacity maximum (P=0.036) were beneficial to be improved in the tai chi group compared to the control group by the analysis of mixed linear model. No adverse effects were found during the intervention period.@*CONCLUSION@#The 12-week tai chi exercise did not have an obvious beneficial effect on cardiac structure, cardiac function and static lung function in older community-dwelling adults at risk of ischemic stroke. The study with a longer duration of intervention should be necessary. (Trial registration No. ChiCTR-TRC-13003601).

Community structure and diversity of soil bacteria in rhizosphere of Salvia miltiorrhiza and Salvia miltiorrhiza f. alba based on pyrosequencing / 中国中药杂志

Rhizosphere bacteria play a vital role in plant nutrition absorption,growth and disease resistance. In this study,high-throughput sequencing technology was used to analyze the rhizosphere bacterial communities of Salvia miltiorrhiza and S. miltiorrhiza f. alba. Moreover,the function of dominant rhizosphere bacterial communities was analyzed. We found that Sphingobacteriales,Sphingomonadales and Nitrosomonadaceae were both dominant and specific bacteria in the rhizosphere of S. miltiorrhiza. The main functions of dominant rhizosphere bacteria communities in both species include promoting transformation of soil nutrients,improving plant immunity and ability of stress tolerance. This study was the first to compare rhizobacterial communities structure and function of S. miltiorrhiza and S. miltiorrhiza f. alba,which provided a new theoretical reference for studing the rhizosphere mechanism of healthy S. miltiorrhiza planting in the future.

Pleiotropic action of genetic variation rs1344706 in ZNF804A on brain structure and function in pa-tients with schizophrenia / 中华行为医学与脑科学杂志

Objective To explore the association between the ZNF804A gene genetic variation poly-morphism rs1344706 and brain structure and function in patients with schizophrenia. Methods Literature search was conducted in Pubmed and other databases,the processes were performed in strict accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines,then the Comprehensive Meta-Analysis software was used for meta-analysis. Results Schizophrenia patients with rs1344706 risk al-lele had lager gray matter in the amount of brain regions including frontal lobe (z＝3.445,P＝0.001),tempo-ral lobe (z＝2.140,P＝0.032) and other brain regions; healthy controls with the risk allele had smaller gray matter and regional activity in the frontal lobe ( gray matter: z＝－2.008, P＝0.045, regional activity: z＝－4.036,P＜0.01) and other regions. Sensitivity analysis was stable,but publication bias existed in a few ana-lyses of indexes. Conclusion The risk allele in ZNF804A gene rs1344706 has positive effects on the brain structure in patients with schizophrenia,but negative effects on the brain structure and function in the healthy individuals.

The structure and function of JMJD2B protein and its progress in the development of malignant tumor / 实用肿瘤学杂志

The JMJD family is an important group of histone demethylase. JMJD2B belongs to one of its family members,and is a JMJD containing a JmjC domain. It mainly regulates chromatin structure,transcription and cell appearance. Recent studies from domestic and foreign have demonstrated that JMJD2B protein is high expressed in human malignant tumors,such as liver cancer,gas-tric cancer,breast cancer,kidney cancer,and skin cancer. Moreover,it also shows that JMJD2B is involved in the occurrence,develop-ment,migration,infiltration,and proliferation of malignant tumors.

Advances in structure-function relation of plant type Ⅲ polyketide synthases by site-directed mutagenesis / 生物工程学报

Plant type Ⅲ polyketide synthases (PKSs), the pivotal enzymes in the biosynthesis of polyketides, produce backbones of many structurally diverse and functionally different polyketides. So far, a variety of functionally diverse plant type Ⅲ PKSs have been cloned and identified from plant origin. Site-directed mutagenesis is a useful technique to study the complex relationship between protein structure and function. This review summarized advances in the structure-function relation of plant type Ⅲ polyketide synthases by site-directed mutagenesis in recent years, including the modification of the amino acid residues influencing enzyme architectures (such as controlling the specificity of starter substrates, the number of condensation reactions, and the cyclization reactions of the intermediate product). This review provides information to study the structure-function relation of plant type Ⅲ polyketide synthases.

Gene floR e a resistência ao florfenicol em isolados de Aeromonas spp. autóctones de organismos aquáticos / Gene floR and resistance to florfenicol in isolated Aeromonas spp. indigenous aquatic organisms

O gene floR descrito é descrito pela literatura como o responsável pela resistência ao florfenicol, que é um antimicrobiano amplamente utilizado na aquicultura. Esse gene já foi relatado em muitas espécies de bactérias, inclusive no gênero Aeromonas. Essas bactérias causam alta mortalidade na piscicultura trazendo prejuízos econômicos. É importante que haja estudos sobre esse gene e possíveis mutações que possam levar a alterações na estrutura e função da proteína. Os objetivos desse estudo foram caracterizar o gene floR em isolados de Aeromonas spp. obtidas do Vale do São Francisco e verificar se a presença desse gene está associada com a resistência ao florfenicol. Foram realizadas reações em cadeia da polimerase (PCR) para a presença do gene floR em 27 isolados de Aeromonas spp.. Amostras positivas para a presença do gene foram sequenciadas e analisadas quanto à presença de polimorfismos por meio de alinhamentos. Os diferentes haplótipos detectados foram utilizados para análises com os programas SIFT e PolyPhen para predição de alteração de função proteica. A modelagem estrutural da proteina codificada pelo gene floR foi realizada com o programa Modeller e, os modelos foram avaliados pelo Procheck, Verify3D e Whatif. A similaridade da estrutura tridimensional da proteína referência com as estruturas tridimensionais das proteínas codificadas pelos diferentes haplótipos foi comparada através do TM-align. A resistência das bactérias ao florfenicol foi avaliada através do teste de microdiluição em caldo, o qual também foi realizado na presença do carbonil cianeto m-clorofenil hidrazona para verificar o efeito da inibição da bomba de efluxo sobre tal resistência. Dos vinte e sete isolados avaliados quanto a presença do gene floR, 14 isolados foram positivos e 10 foram sequenciados, o que permitiu a identificação de três polimorfismos no gene floR, que levaram a construção de três haplótipos diferentes (TAA, TTA e CTG). As análises realizadas com os programas SIFT e PolyPhen apontaram que os haplótipos TTA e TAA provavelmente poderiam alterar a estrutura e função da proteína. As proteínas modeladas para os três haplótipos demonstraram possuir praticamente a mesma conformação estrutural entre si. Todos os isolados que apresentaram o gene foram resistentes ao florfenicol e aqueles que não apresentavam foram sensíveis. O teste na presença do Carbonil Cianeto m-Clorofenil Hidrazona foi realizado para três isolados, cada isolado representando um haplótipo, sendo possível observar a inibição do crescimento bacteriano em todas as concentrações independente do haplótipo. Os resultados obtidos nesse estudo mostram que a resistência ao flofenicol em Aeromonas spp. pode ser explicada pela presença do gene floR, e que esse gene está relacionado com uma bomba de efluxo. As mutações verificadas no gene floR, parecem não estar envolvidas com alteração de estrutura e função da proteína codificada por esse gene.(AU)

The floR gene is described in related literature as responsible for resistance to florfenicol, which is a widely used antimicrobial agent in aquaculture. This gene has been reported in many species of bacteria, including the genus Aeromonas. These bacteria cause high mortality in fish farming bringing economic losses. It is important that studies of this gene and possible mutations that can lead to changes in the structure and function of the protein. The aim of this study was to characterize the floR gene in isolates of Aeromonas spp. and check if the presence of this gene is associated with resistance to florfenicol in Aeromonas spp. obtained from the San Francisco Valley. PCR (Polymerase Chain Reaction) were also performed to verify the presence of the floR gene in 27 isolates of Aeromonas spp. Positive samples for the presence of the gene were sequenced and analyzed for the presence of polymorphisms using alignments. Different haplotypes detected were used for analysis with the SIFT and PolyPhen programs for prediction of changes in protein function. The structural modeling of protein encoded by the floR gene was performed using the Modeller software, and the models were evaluated by Procheck, Verify3D and Whatif. The similarity of the dimensional structure of reference protein with the dimensional structures of the proteins encoded by the different haplotypes was compared by TM-align. Bacterial resistance to florfenicol was assessed by the microdilution test, which was also performed in the presence of carbonyl cyanide m-chlorophenyl hydrazone to verify the effect of inhibiting the efflux pump. 14 isolates were positive for the presence of floR gene and 10 were sequenced and allowed the identification of three polymorphisms in the floR gene, which led to construction of three different haplotypes (TAA TTA and CTG). The analyzes carried out with the SIFT and PolyPhen programs showed that the TTA and TAA haplotypes could probably change the protein structure-function. Proteins modeled for the three haplotypes were found to have substantially the same structural conformation with each other. All isolates presenting the gene were resistant to florfenicol and those who did not have were sensitive. The test in the presence of carbonyl cyanide m-chlorophenylhydrazone was conducted for three isolates, representing each single haplotype and was observed inhibition of bacterial growth at all concentrations independent of the haplotype. The results of this study show that resistance to flofenicol in Aeromonas spp. may be explained by the presence of floR gene and that this gene is associated with an efflux pump. Mutations observed in floR gene do not appear to be involved with chenges in structure and function of the protein encoded by gene.(AU)

Gene clone, eukaryotic expression vector and tissue expression profile analysis of porcine BST-2 gene / 中国兽医学报

In order to study the biological function of pig BST-2 gene,the BST-2 gene was amplified with specific primers from porcine kidney tissue,and molecular characterization of BST-2 nuclectide and amino acid sequence were analyzed with bioinformatics tools and online server.Then the prokaryotic expression and tissue expression profile analysis was carried out.The results showed that the full length of pig BST-2 gene was 851 bp and contained 23 bp of 5'-UTR,294 bp of 3'-UTR and 534 bp of CDS and the gene encoded 177 aa.Amino acid sequence analysis of pig BST-2 protein showed 46.1％ identity with gorilla gorilla,41.7％ with cricetulus griseus,39.5％ with mus musculus,35.4％ with equus asinus,42.0％ with felis catus,40.5％ with bos mutus,44.4％ with macaca mulatta,38.7％ with ovis aries and 46.8％ with homo sapiens.BST-2 protein contained 2 transmembrane structure (27-49 aa and 154-176 aa),2 glycosylation sites and 14 potential phosphorylation sites including ATM,CK Ⅱ,PKA,PKC binding sites.The pig BST-2 protein was expressed in Vero cells after translated the recombinant plasmid FLAG-BST-2.Semiquantitative PCR results showed that BST-2 gene was expressed in all the tissues,especially in lymph nodes,thymus,tonsils,spleen,large intestine and small intestine.This study provide a foundation for further understanding the antiviral mechanism of pig BST-2 protein.

Effect of aerobic exercise on pancreatic islets and explorationof their mechanism in type 2 diabetic rats / 中国比较医学杂志

Objective To explore the effect and mechanism of aerobic exercise on islet β-cells in type 2 diabetic rats.Methods Thirty healthy SPF 8-week old Wistar rats were randomly divided into control group (C, n=10), diabetic control groups (DMC) and diabetic exercise (DME) groups, 10 rats in each group, among which 7 successful rat models were used in the experiment.The diabetic rat model was established by high fat and sugar diet and i.p.injection of streptozotocin in a dose of 50 mg/kg.The rats of group DME were forced to perform 20 m/min running for 30 min, once a day, 6 days in a week, for 8 weeks.Other rats were allowed free movement.At the end of experiment, serum glucose and insulin were measured and homeostasis model assessment (HOMA) was calculated, and pancreatic tissue samples were collected for histopathological examination.The morphology and structure of pancreatic islets were observed under a digital microscope, the perimeter and area of islets were analyzed by image analysis, and shape factor of islets was calculated.The insulin content, glucokinase and ultramicro-ATPase activity in the pancreatic homogenate were determined.Results In the DME group, the perimeter and area of islets were significantly higher than the DMC group (P< 0.05), but still lower than the control group.The shape factor was significantly increased, the cell hypertrophy, vacuolization and nuclear pyknosis were markedly alleviated than those in the DMC group, the insulin content, glucokinase and the trace total ATP activity in the DME group were significantly higher than those in the DMC group (P<0.05), and the SF and HOMA were significantly changed.Conclusions Aerobic exercise can reduce the blood glucose level, improve the morphology of islets and β-cells in the type 2 diabetic rats.It may be due to increase of the activity of glucose kinase and ATP-synthase, and increased insulin sensitivity in the pancreas.

Impairment of renal structure and function following heterogeneous chemical mixture exposure in rats.

Renal structural and functional alterations following an exposure to a heterogeneous chemical mixture (HCM) of phthalic acid di butyl ester, 1, 2–dichlorobenzene, cadmium chloride and chromium trioxide, administered through oral gavage in low doses (1/100 and 1/1000 of LD50 value of individual chemical) for 60 days, followed by withdrawal till 120 days resulted in significant rise in kidney lipid peroxidation and fall in the activities of enzymatic antioxidants. However, withdrawal of HCM treatment restored most of these altered parameters. Degenerative changes in the kidney included proximal convoluted tubules devoid of brush boarder with cytoplasmic blebbing, dissolution and sloughing of nuclei. Cortical glomeruli were also affected with epithelial disintegration, pyknosis of podocyte nuclei and mesengial cell hyperplasia. The morphological alterations recovered fully in the low dose compared to the high dose treatment group.

Identification and characterization of a gene encoding a putative lysophosphatidyl acyltransferase from Arachis hypogaea.

Lysophosphatidyl acyltransferase (LPAT) is the important enzyme responsible for the acylation of lysophosphatidic acid (LPA), leading to the generation of phosphatidic acid (PA) in plant. Its encoding gene is an essential candidate for oil crops to improve oil composition and increase seed oil content through genetic engineering. In this study, a fulllength AhLPAT4 gene was isolated via cDNA library screening and rapid amplification of cDNA ends (RACE); our data demonstrated that AhLPAT4 had 1631 nucleotides, encoding a putative 43.8 kDa protein with 383 amino acid residues. The deduced protein included a conserved acyltransferase domain and four motifs (I–IV) with putative LPA and acyl-CoA catalytic and binding sites. Bioinformatic analysis indicated that AhLPAT4 contained four transmembrane domains (TMDs), localized to the endoplasmic reticulum (ER) membrane; detailed analysis indicated that motif I and motifs II–III in AhLPAT4 were separated by the third TMD, which located on cytosolic and ER luminal side respectively, and hydrophobic residues on the surface of AhLPAT4 protein fold to form a hydrophobic tunnel to accommodate the acyl chain. Subcellular localization analysis confirmed that AhLPAT4 was a cytoplasm protein. Phylogenetic analysis revealed that AhLPAT4 had a high homology (63.7–78.3%) with putative LPAT4 proteins from Glycine max, Arabidopsis thaliana and Ricinus communis. AhLPAT4 was ubiquitously expressed in diverse tissues except in flower, which is almost undetectable. The expression analysis in different developmental stages in peanut seeds indicated that AhLPAT4 did not coincide with oil accumulation.

Multivariant analysis for structural and functional changes of left ventricle in patients with lupus nephritis / 中华风湿病学杂志

Objective To investigate the correlation between the changes of left ventricle and age,duration,blood-pressure,hemoglobin,serum creatinine level and glomerular filtration rate (GFR) in lupus nephritis (LN) patients.Methods One hundred and thirty-two patients with systemic lupus erythematosus (SLE) were divided into LN group and non-LN group,the clinical data were analyzed with t test,x2 test,Pearson correlation analysis and multiple regression analysis.Results Compared with non-LN patients (4/67,6％ ),the incidence of concentric hypertrophy was significantly increased in LN patients ( 14/65,22％ ) (x2=6.790,P＜0.05 ).Left ventricular end systolic internal dimension (LVESd) was correlated positively with blood uric acid (BUA) level (B=0.014,P＜0.01 ).Left ventricular mass index (LVMI) was correlated positively with blood urea nitrogen (BUN) level (B=2.977,P＜0.01 ).lnter-ventricular septal thickness (IVST) was correlated positively with systolic blood pressure (B=0.022,P＜0.01 ).Conclusion Compared with non-LN patients,the incidence of concentric hypertrophy is significantly increased in LN patients.It may be one of the mechanisms that SLE could involve kidney,which consequently lead to hypertension,higher BUN and BUA level,which may further aggravate heart damage in turn.