ABSTRACT
Adenosine diphosphate (ADP)-ribosylation is a unique post-translational modification that regulates many biological processes, such as DNA damage repair. During DNA repair, ADP-ribosylation needs to be reversed by ADP-ribosylhydrolases. A group of ADP-ribosylhydrolases have a catalytic domain, namely the macrodomain, which is conserved in evolution from prokaryotes to humans. Not all macrodomains remove ADP-ribosylation. One set of macrodomains loses enzymatic activity and only binds to ADP-ribose (ADPR). Here, we summarize the biological functions of these macrodomains in DNA damage repair and compare the structure of enzymatically active and inactive macrodomains. Moreover, small molecular inhibitors have been developed that target macrodomains to suppress DNA damage repair and tumor growth. Macrodomain proteins are also expressed in pathogens, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, these domains may not be directly involved in DNA damage repair in the hosts or pathogens. Instead, they play key roles in pathogen replication. Thus, by targeting macrodomains it may be possible to treat pathogen-induced diseases, such as coronavirus disease 2019 (COVID-19).
Subject(s)
Humans , ADP-Ribosylation , COVID-19/metabolism , DNA Repair/physiology , Evolution, Molecular , Models, Biological , Models, Molecular , N-Glycosyl Hydrolases/metabolism , Poly(ADP-ribose) Polymerases/metabolism , Protein Domains , SARS-CoV-2/pathogenicityABSTRACT
Objective:In order to improve the understanding of the clinical, endoscopic and pathological characteristics of occult primary intestinal lymphangiectasia in infants, and to reduce misdiagnosis and missed diagnosis, we analyzed the clinical manifestations and diagnosis and treatment related data of a case of primary intestinal lymphangitis in Xiamen Children′s Hospital.The patient was admitted to the hospital with bronchopneumonia, mild diarrhea and edema, without decreasing in the number of peripheral blood lymphocytes, showing hypoproteinemia.Albumin infusion failed to alleviate hypoalbuminemia, and then gastroscopy showed that the duodenal segment had white millet like granular protrusion with different sizes and dense density, and the mucosa had extensive leukoplakia like lesions.Pathological examination showed that the small intestinal lymphadenectasia was obvious, and then the small intestinal lymphadenectasia was confirmed.After diagnosis, albumin was infused every two days for two consecutive courses and portagen medium chain fatty acid milk powder was fed.After treatment, the total protein and albumin in the plasma basically rose to the normal level, and the color Doppler ultrasonography of abdomen and chest showed that both pleural effusion and bilateral pulmonary effusion had been absorbed.The retrospective analysis of this case shows that for infants with dropsy caused by the decrease of plasma albumin and globulin, the possibility of excessive protein consumption caused by liver and kidney disease, tumor, tuberculosis and other chronic diseases should be excluded, and small intestinal lymphadenopathy should be considered, gastroscopy should be performed as soon as possible, pathological biopsy should be further improved, early diagnosis and treatment should be carried out as soon as possible And diet intervention treatment to avoid serious consequences caused by not timely handling.
ABSTRACT
OBJECTIVE:To establish HPLC fingerprint of Qingbi granules.METHODS:HPLC method was adopted.The determination was performed on Diamonsil C18 column with mobile phase consisted of methanol-0.3% phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 260 nm,and column temperature was 25 ℃.The sample size was 10 μL.Using baicalin as reference,HPLC chromatograms of 10 batches of samples were determined.Common peak identification and similarity evaluation were performed by using TCM Chromatographic Fingerprint Similarity Evaluation Software (2.0 edition).RESULTS:There were 29 common peaks in HPLC chromatograms of 10 batches of samples.The similarity among the 10 batches was more than 0.90.After validation,HPLC chromatograms of 10 batches of samples were in line with control fingerprints.CONCLUSIONS:Established fingerprints can provide reference for identification and quality evaluation of Qingbi granules.
ABSTRACT
OBJECTIVE:To study the effect of EGCG-Zn chelated by epigallocatechin gallate (EGCG) and Zn ion on learn-ing,memory and antioxidant abilities in vascular dementia(VD)rats. METHODS:Rats were randomly divided into sham group, model group,zinc gluconate group (positive control,70 mg/kg),EGCG group (5 mg/kg),and EGCG-Zn low-dose,mid-dle-dose,high-dose groups (2,5,10 mg/kg),10 in each group. The VD models were induced by middle cerebral artery occlu-sion. The rats were intragastrically given relevant medicines after 7 d of modeling,once a day,for 28 d. Morris water maze were used to evaluate learning and memory abilities of rats. Spectrophotometric method was adopted to detect the superoxide dismutase (SOD),catalase(CAT),glutathione peroxidase(GSH-Px)activities and malondialdehyde(MDA)content in brain tissue of rats. RESULTS:Compared with sham group,escape latency was extended in model group,percentage of platform quadrant swimming distance(dP/dT)was reduced(P<0.01);SOD,CAT,GSH-Px activities in brain tissue were reduced,and MDA content was in-creased (P<0.01). Compared with model group,escape latency on 4th d of recording was obviously shortened in EGCG group and EGCG-Zn dose groups,dP/dT was obviously increased and dose-depended(P<0.05 or P<0.01);SOD,CAT,GSH-Px activi-ties in brain tissue were increased,MDA content was decreased(P<0.05 or P<0.01)and dose-depended. Compared with EGCG group and zinc gluconate group,escape latency was obviously shortened in EGCG-Zn dose groups,dP/dT was increased;SOD, CAT,GSH-Px activities in brain tissue were increased,MDA content was decreased;and the effects in high-dose,medium-dose groups were more obvious(P<0.05 or P<0.01). CONCLUSIONS:EGCG-Zn can improve the learning,memory and antioxidant abilities in VD rats,and the effect is superior to EGCG.
ABSTRACT
Objective:To evaluate the biological effects of Luteolinl on the proliferation and differentiation in human periodontal ligament cells (PDLC) . Methods:MTT, ALP kit and Q-PCR was used to detect the expression of osteogenesis related gene. Results:Luteolinl (100, 10, 1, 0.1, 0.01μmol/L) could increase the proliferation of PDLC, and there were significant differences compared with control group (P<0.05) . The ALP Kit results showed that Luteolinl could increase the ALP actiuty of PDLC (P<0.05) . The Q-PCR results showed that Luteolin could increase the expression of ALP and RUNX2 (P<0.05) . Conclution:At proper concentration,Luteolinl can increase the proliferation and differentiation of PDLC.
ABSTRACT
Vascular smooth muscle cells (VSMCs) proliferation is the critical pathological process of in-stent restenosis (ISR). Recent researches indicate that the traditional Chinese medicine curcuma zedoary (E’zhu) can inhibit VSMCs proliferation, with a potential value in preventing and treating ISR. The major ac-tive components of curcuma zedoary are curcuma and β-elemene. The underlying mechanisms involve the inhibition of hemeoxygen-ase-1 expression, blockade of extracellular signal-regulated ki-nase – MAPK and Akt pathways, and subsequent cell cycle ar-rest. This article reviews the recent progress on curcuma zedoary extracts regulating VSMCs proliferation in ISR.
ABSTRACT
Objective To establish the quality standards for compound Shatagan Oral Liquid. Methods Chuanxiong Rhizoma was identified by TLC. The content of ferulic acid was determined by HPLC. Separation was performed on a Diamonsil C18 column (4.6 mm × 250 mm, 5μm) with a mobile phase consisting of methanol-1% acetic acid solution in gradient elution (0-5 min, 35% methanol;5-8 min, 35%→23% methanol;8-22 min, 23% methanol) at 30℃;The flow rate was 1.0 mL/min;The injection volume was 5μL;The detection wavelength was 322 nm.Results Ferulic acid showed a good linear relationship in the range of 0.039 4-0.630 0μg (r=0.999 7,n=7). The average recovery was 98.22% and RSD was 2.62% (n=6).Conclusion The method is reliable, sensitive and with repeatability, which can be used as the quality control method for compound Shatagan Oral Liquid.
ABSTRACT
Background It is determined that riboflavin/ultraviolet A (UVA)-induced corneal collagen crosslinking is able to increase resistance of cornea against enzymatic digestion and has antimicrobial efficacy for various kinds of bacteria in vitro.However,its in vivo study is less now.Objective This study aimed to evaluate the efficacy of iontophoresis-mediated corneal collagen crosslinking combined with or without drugs for Staphylococcus aureus keratitis.Methods Bacterial keratitis models were induced by the interstromaly injection of Staphylococcus aureus suspension with concentration 2× 109/ml in the right eyes of 40 rabbits,and then the rabbits were randomly classified into the model group,gatifloxacin eye drops group,riboflavin/UVA corneal crosslinking group and drugs+ crosslinking group.The smearing of corneal surface was performed for the identification of bacteria 24 hours after injection.Iontophoresis-mediated riboflavin/UVA crosslinking was applied on the eyes of the riboflavin/UVA corneal crosslinking group and drugs+crosslinking group,and gatifloxacin eye drops was topically used 7 times per day on the eyes of the gatifloxacin eye drops group and drugs+crosslinking group.The corneal inflammation was examined and graded under the slit lamp biomicroscope before and after treatment.Ocular anterior segment optical coherence tomography(AS-OCT),corneal histopathology and ultrastructure were examined 14 days after treatment.The living environment of the experimental animals was maintained at 21 ℃ with a 12-hour light and dark cycle.Animals used in this study were treated in accordance with the Weifang Medical College Animal Experimentation Ethic Committee (AEEC) guidelines.The study protocol was approved by the AEEC.Results Corneal inflammation and ulcer were observed,but no significant difference was found in the inflammatory grade among the 4 groups 24 hours after injection (x2=0.293,P>0.05).In the 14th day after injection,the corneal ulcer area was smaller and corneal edema was milder in the drugs+crosslinking group compared with the model group,gatifloxacin eye drops group and riboflavin/ UVA corneal crosslinking group,showing a significant difference in the inflammatory grade among them (x2 =38.710,P<0.001).The cornea thickness values of ulcer zone were (428.1 ± 146.2) μm on the 14th postinjected day in the drugs+crosslinking group,which was evidently higher than those in the model group,gatifloxacin eye drops group and riboflavin/UVA corneal crosslinking group,with a significant difference among the 4 groups (F =8.310,P<0.001).A lower degree of destruction of cornea collagen and less inflammatory cells were seen in the cornea tissue of the drugs+ crosslinking group by haematoxylin and eosin staining in comparison with other 3 groups,and normal keratocytes were much more in the drugs + crosslinking group than those in other treated groups.Conclusions Iontophoresismediated corneal collagen crosslinking can alleviate Staphylococcus aureus keratitis.The combination of crosslinking with drugs has a better effectiveness than the administration of gatifloxacin eye drops only or riboflavin/UVA corneal crosslinking only.
ABSTRACT
AIM:To investigate whether mitochondrial membrane potential (ΔΨm ) and the mitochondrial ap-optotic pathway are involved in the protective mechanism of Panax quinquefolium saponin ( PQS) against cardiomyocyte ap-optosis after ischemia/reperfusion ( I/R) injury in rat myocardium .METHODS: Ninety healthy male SD rats were ran-domly divided into sham group , I/R group, PQS (200 mg· kg -1 · d-1 ) +I/R group, cyclosporine A ( CsA) group, CsA (10 mg· kg-1 ) +I/R group and PQS +CsA +I/R group.The model of myocardial I/R injury in vivo was established by ligating the left anterior descending artery ( LAD) for 30 min followed by 120 min of reperfusion in the rats .The serum ac-tivity of lactate dehydrogenase ( LDH ) was measured by automatic chemistry analyzer .The myocardial infarct size was measured by Evans blue and 2,3,5-triphenyltetrazolium chloride (TTC) staining.Cardiomyocyte apoptosis was detected by in situ TDT-mediated dUTP nick end labeling (TUNEL).The protein levels of Bcl-2, Bax, cleaved caspase-3 and cytosol-ic cytochrome C were determined by Western blotting .ΔΨm was measured by laser scanning confocal microscopy and fluo-rescence microplate reader .RESULTS:Compared with I/R group, the serum content of LDH ,the infarction size in PQS+I/R group, CsA+I/R group and PQS +CsA+I/R group and the myocardial apoptotic index were decreased .Compared with I/R group, the fluorescence intensity of mitochondria after JC-1 staining was enhanced in PQS +I/R group, CsA+I/R group and PQS+CsA+I/R group, and the relative fluorescence units (RFU) ofΔΨm were improved in those 3 groups. In PQS+I/R group, CsA+I/R group and PQS+CsA+I/R group, the protein expression of Bcl-2 was increased, and that of Bax was decreased compared with I/R group.Moreover, in those 3 groups, the protein levels of cleaved-caspase-3 and cytosolic cytochrome C were decreased compared to I /R group, respectively.CONCLUSION:PQS attenuates myocardial injury and cardiomyocyte apoptosis during I /R, and the protective mechanisms of PQS were associated with the modulation ofΔΨm and the inhibition of mitochondrial apoptosis pathway .
ABSTRACT
AIM:To study the effect of Panax quinquefolium saponin (PQS) on cardiomyocyte apoptosis in-duced by thapsigargin ( TG) .METHODS:Primary cultured cardiomyocytes from neonatal SD rats were divided into con-trol group, TG group, PQS (40 mg/L, 80 mg/L and 160 mg/L) +TG group, si-PERK+TG group, and mock+TG group.The cells were treated with 1 μmol/L TG for 24 h to induce apoptosis.The PERK gene in the cardiomyocytes was knocked down by RNAi.The cell viability was detected by CCK-8 assay.Apoptosis was analyzed by flow cytometry.Wes-tern blotting was used to determine the expression of ERS molecules GRP78, CRT, ATF4 and CHOP, anti-apoptosis pro-tein Bcl-2 and pro-apoptosis protein Bax.RESULTS: Compared with control group, TG significantly and the apoptosis, reduced the cell viability (P<0.05), increased the phosphorylation of PERK and eIF2α, increased the expression of GRP78, CRT, ATF4, CHOP and pro-apoptosis protein Bax, and decreased the expression of anti-apoptosis protein Bcl-2 (P<0.05).Compared with TG group, PQS treatment (160 mg/L) significantly reduced the apoptosis and increased the cell viability (P<0.05).All the 3 different concentrations of PQS significantly increased the expression of anti-apoptosis protein Bcl-2 and reduced the expression of pro-apoptosis protein Bax (P<0.05) in a dose-dependent manner.PQS pre-treatment and knockdown of PERK both reduced the protein levels of GRP78, CRT, PERK, p-PERK, eIF2α, p-eIF2α, ATF4, CHOP and pro-apoptosis protein Bax, and increased the expression of anti-apoptosis protein Bcl-2 (P<0.05). CONCLUSION:PQS at concentration of 160 mg/L attenuated cardiomyocyte apoptosis induced by TG.PQS had the simi-lar effect as PERK knockdown on cardiomyocyte apoptosis.The mechanism may be associated with inhibiting PERK-eIF2α-ATF4-CHOP pathway of ERS-related apoptosis.
ABSTRACT
Objective To observe the effects of resveratrol on blood pressure and cardiac function in the rats with vascular calcification induced by vitamin D3 plus nicotine.Methods 32 male SD rats were randomly divided into the following groups for treatment:Control (Con), calcified (Cal), Cal+Res low dose [L] and Cal+Res high dose [H] groups.Blood pressure, cardiac function, alkaline phosphatase ( ALP ) activity in serum or aorta were detected and HE staining was used for pathological examination at 6 weeks after treatment.Results Compared with the Con group, LVW/BW, heart rate, systolic aortic pressure, pulse pressure, mean blood pressure, left ventricular systolic pressure and ALP activity in serum or aorta of rats in the Cal group were increased by 27.3%, 8.8%, 22.8%, 47.5%, 13.6%, 19.0%, 280%and 265%( P<0.05 or P<0.01) , respectively.Compared with the Cal group, pulse pressure and ALP activity in serum or aorta of rats in the Cal+Res [L] group were decreased by 8.5%, 34.5%and 29.5%(P<0.05 or P<0.01), respectively, and LVW/BW, systolic aortic pressure, pulse pressure, mean blood pressure, left ventricular systolic pressure and ALP activity in serum or aorta of rats in the Cal +Res [ H] group were decreased by 14.2%, 13.6%, 23.7%,10.0%, 9.0%, 53.1%and 45.9%(P<0.05 or P<0.01), respectively.Compared with the Cal+Res [L] group, systolic aortic pressure, pulse pressure, left ventricular systolic pressure and ALP activity in serum or aorta of rats in the Cal+Res [H] group were decreased by 8.3%, 16.7%, 5.8%, 28.4% and 23.2% (P<0.05 or P<0.01), respectively.Compared with the aorta in the Con group, pathological examination revealed thickened vessel walls and disordered elastic fibers in the calcified aortas.However, the thickness of aortic wall in the Cal+Res [ L] and Cal+Res [ H] groups was reduced and elastic fibers were regularly arranged.Conclusion Resveratrol can effectively reduce the blood pressure and improve the cardiac function in rats with vascular calcification.
ABSTRACT
Further study on the chemical constituents of Artemisia sphaerocephala led to the isolation of fifteen compounds. These compounds were isolated and purified by repeated column chromatography on silica gel and Sephadex LH-20. Their structures were determined by physicochemical properties and spectral data analysis. These compounds were identified as 5-hydroxy-7,4'-dimethoxyflavanone (1), 5-hydroxy-7, 4'-dimethoxyflavone (2), 5-hydroxy-7, 4'-dimethoxyflavanonol (3), 5,3'-dihydroxy-7, 4'-dimethoxyflavanone (4), 5,7-dihydroxy-6,4'-dimethoxyflavone (5), isosakuranetin (6), hesperetin (7), navingenin (8), acacetin (9), chrysoeroil (10), 5,7-dihydroxy-4'-methoxyflavone-6,8-di-C- -glucopyranoside (11), didymin (12), acacetin-7-O-rutinoside (13), piceine (14), and capillarin (15). Compounds 1-3, 5, 7, 8, 10-15 were isolated from this plant for the first time.
Subject(s)
Artemisia , Chemistry , Chromatography , Methods , Flavones , Flavonoids , Glucosides , Plant ExtractsABSTRACT
Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was aimed to find out whether these miRNAs participate in regulating myocardial ischemia reperfusion (I/R) injury.Methods Apoptosis in mice hearts subjected to I/R was detected by TUNEL assay in vivo,while flow cytometry analysis followed by Annexin V/PI double stain in vitro was used to detect apoptosis in cultured cardiomyocytes which were subjected to hypoxia/reoxygenation (H/R).Taqman real-time quantitative PCR was used to confirm whether miR-15a/15b/16 were involved in the regulation of cardiac I/R and H/R.Results Compared to those of the controls,I/R or H/R induced apoptosis of cardiomyocytes was significantly iucreased both in vivo (24.4% ± 9.4% vs.2.2% ± 1.9%,P < 0.01,n =5) and in vitro (14.12% ±0.92% vs.2.22% ± 0.08%).The expression of miR-15a and miR-15b,but not miR-16,was increased in the mice I/R model,and the results were consistent in the H/R model.Conclusions Our data indicate miR-15 and miR-15b are up-regulated in response to cardiac I/R injury,therefore,down-regulation of miR- 15a/b may be a promising strategy to reduce myocardial apoptosis induced by cardiac I/R injury.
ABSTRACT
ObjectiveTo observe nailfold capillary changes in a cohort of conncctive tissue disease ( CTD ) with Raynaud's phenomenon (RP) and to explore the diagnostic value of nailfold videocapillaroscopy (NVC) in systemic sclerosis(SSc).MethodsSixty CTD patients with RP divided into SSc group (n =36) and non-SSc group ( n =24) were referred to an experienced operator for NVC.ResultsThe patients had decreased capillary loops in SSc group with the capillary diameter more enlarged in SSc group than nonSSc group.The number of patients in SSc group with giant capillaries was 14,while 3 in non-SSc group.There were 23 patients with haemorrhagcs in SSc group and 9 in non-SSc group.The number of patients with severe effusion was 15 in SSc group,while 2 in non-SSc group.By using the ROC curves,indexes with AUC at least 0.7 of the input capillary diameter,the output capillary diameter,the middle capillary diameter,blood color and effusion for the diagnostic cutoff points were 18.5 μm,24.5 μm,19.5μm,deep red and severe effusion.With at least 2 out of the top 3 indexes,the diagnostic sensitivity and specificity of SSc were higher.ConclusionsCTD Patients with RP of SSc have less capillary loops,more enlarged capillaries,more giant capillaries, moresevereeffusionandmorehaemorrhagesthannon-SScpatients. The characteristics of nailfold capillary changes in SSc patients with RP can be helpful tor the diagnosis and the differential diagnosis of SSc.
ABSTRACT
BACKGROUND: Silk, a natural product, has been paid wide attention in medical field owing to a fact that silk's mechanical property and biocompatibility are superior to traditional artificially synthesized degradable polymers. OBJECTIVE: To observe the effects of silk on the adsorption as well as cell morphology and function of 3T3-L1 preadipocytes. METHODS: Raw silk and trypsin-digested silk were ad Ubitum twisted into reticular fibrous cord with three-dimensional structure: mesh size 70-200 μm, thickness 200-300 μm, and porosity 20%. The three-dimensional scaffolds made of digested silk were placed in the 24-well plate. 3T3-L1 preadipocytes suspension (6×10~(10)/L) was added to the culture plate, 3 drops comprising 1 × 10~7 cells per well. Following 4-hour incubation in the air, when cells fully attached to the scaffold, they were thoroughly soaked with medium, which was renewed every other 2-3 days for a total of 1-4 weeks.RESULTS AND CONCLUSION: Inverted microscope results showed that silk scaffold/3T3-L1 preadipocytes compounds presented with slender prominences that stretched out and migrated ahead to gradually connect together through a head-to-end fusion fashion and enter into meshes. Scanning electron microscope results demonstrated that in the silk scaffold/3T3-L1 preadipocytes compounds, ceils tightly attached to silk scaffold, appropriately spread out, and secreted matrix. Silk shows better absorption for 3T3-L1 preadipocytes and maintains the normal morphology and functions of 3T3-L1 preadipocytes.
ABSTRACT
AIM: To prepare and purify the polyclonal antibodies against human myofibrillogenesis regulator 1 (hMR-1), then to characterize the purity, titer, specificity and the availability.METHODS: Two polypeptides named peptide 1 and 2 were synthesized based on the bioinformatics analysis of the sequence of hMR-1 by using software TMHMM and DNAStar, then coupled with keyhole limpet hemocyanin (KLH) for immunization. These peptides for immunization were mixed and injected into New Zealand rabbits to prepare antibodies specifically against hMR-1. ELISA assay was used to detect the titers of the antibodies. After purification by immunoaffinity chromatography, antibodies were identified by Western blotting and immunocytofluorescent assays. Applications of the antibodies on neonatal rat cardiomyocytes were also employed.RESULTS: (1)The titers of antibodies were 1:10~5. In WB assay, a specific 17kD band was detected, corresponding to the predicted molecular weight of hMR-1; the positive fluorescent signals were distinct. (2)On the neonatal rat cardiomyocytes model, we observed a peri-nucleus location. The fluorescent signal of hMR-1 overexpression group was much stronger than that in vector control and normal control groups.CONCLUSION: All these results indicate that the antibodies obtained from poly peptides mixture immunization have either human original or rat original antigens. The antibody is available for using in Western blotting or immunofluorescent assays.
ABSTRACT
<p><b>OBJECTIVE</b>To study the accumulation and translocation of cadmium in the soil and Artemisia annua, and observe its effects on growth of A. annua and artemisinin content.</p><p><b>METHOD</b>A. annua were cultivated in pots with Cd concentration at 0.5, 1.5, 4.5 mg x kg(-1) level, respectively.</p><p><b>RESULT AND CONCLUSION</b>The growth of A. annua was inhibited at all the Cd levels characterized by the decreases of biomass and agronomic parameters; Most of Cd was accumulated in the roots of A. annua, and the ratios of Cd concentrations in roots and aerial part were 1.8:1 and 2.3:1 at 1.5, 4.5 mg x kg(-1) Cd level, respectively. Artemisinin content increased significant at 0.5 mg x kg(-1) Cd level, but there were no significant changes comparing with control group other Cd levels.</p>
Subject(s)
Artemisia annua , Chemistry , Metabolism , Artemisinins , Metabolism , Cadmium , Metabolism , Toxicity , Plant Extracts , Metabolism , Soil Pollutants , Metabolism , ToxicityABSTRACT
OBJECTIVE To investigate the correlated resistant genes encoding ?-lactamases,aminoglycoside and genetic marker of integron and transposon in multi-resistant Pseudomonas aeruginosa(MRPA) isolated from clinical specimens and study their relationship by phylogenetic analysis.METHODS Twenty-one resistant genes,two integron-Ⅰ genes and one transposon genetic gene were analyzed by PCR and verified by DNA sequencing.Multi-resistant genes cluster analysis was performed by UPGMA.RESULTS The positive rates of CARB,oprD2,aac(3)-Ⅱ,aac(6′)-Ⅱ,ant(2″)-Ⅰ,intⅠ1,qacE△1-sul1 and merA in 20 strains of MRPA were 15%,100%,70%,15%,15%,85%,85% and 85%,respectively,and other genes were negative.It was classfied to three subgroup,by the multi-resistant genes cluster analysis.CONCLUSIONS MRPA isolated from clinical specimens has carried many resistant genes.The deficiency rate of oprD2 gene is very high.The positive rate of genetic mark genes about integron and transposon is very high.It may be the main multi-resistant mechanism of MRPA.Multi-resistant genes cluster analysis shows that there is clone transmission in MRPA and it can induce nosocomial infection prevalence.
ABSTRACT
@#Objective To observe the effect of Edaravone pharmacological postconditioning and ischemic postconditioning on acute myocardial ischemia and reperfusion injury.Methods 40 rats were randomly divided into the sham operation group,ischemia/reperfusion group,ischemic postcondition group,Edaravone group A(injected through artery before reperfusion)and Edaravone group V(injected through vein before reperfusion)with 8 animals in each group.The animal model of acute myocardial ischemic and reperfusion was established.ECG changes were monitored during the procedure,dynamic parameters and serum biochemical markers creatine kinase MB(CK-MB),malondialdehyde(MDA),superoxide dismutase(SOD)of different groups were assessed and evaluated at the end of reperfusion.Ischemic and infarct areas were measured by Evans blue and TTC staining respectively.Results The myocardial infarct area and levels of serum CK-MB,MDA all significantly reduced(P<0.01)and activity of SOD enhanced(P<0.05)in the ischemic postcondition group and Edaravone group A compared with the ischemia/reperfusion group.The myocardial infarct area and levels of serum CK-MB,MDA reduced and activity of SOD enhanced(P<0.05)in the Edaravone group V compared with the ischemia/reperfusion group.There were no statistical differences in the foregoing indexes among the ischemic postcondition group,Edaravone group A and Edaravone group V group(P>0.05).Conclusion Edaravone injected through artery just before the onset of coronary reperfusion can reduce myocardial infarct area,which is similar to the cardio protective effect of mechanical postconditioning.The common potential mechanism of them might be associated with decreasing the injury by reactive oxygen species and strengthening the resistance to oxidation stress.The effect of intra aorta root-coronary injection is not worse than that of intra vein
ABSTRACT
BACKGROUND:In chronic congestive heart failure (CHF),both protein expression and activity of sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA2a) are decreased, which results in abnormal regulation of calcium ion and systole and diastole dysfunction.OBJECTIVE:To observe the short-and middle-term therapeutic effects of adeno-associated viral gene transfer of SERCA2a in treatment of CHF in rats,and to investigate the possible mechanisms.DESIGN:Randomized-controlled design.SETTING:Department of Geriatric Cardiology and Department of Pathophysiology,General Hospital of Chinese PLA.METHODS:Male Sprague-Dawiey rats,were randomized into 4 groups:sham-operation group,CHF group,CHF+ green fluorescent protein (GFP) group and CHF+SERCA2a group. Gene transfer 10 and 30 days two time points were set in each group.CHF rat models were established by coarctation of abdominal aorta.At postoperative 18 to 20 weeks,the successful rats in CHF group were intraperitoneally injected with 500 μL aseptic normal saline,and those in the CHF+ GFP group and CHF+ SERCA2a group were injected with the same dose of Adeno-associated viral gene with GFP and SERCA2a gene respectively.MAIN OUTCOME MEASURES:①At 10 and 30 days after gene transfer,hemodynamic parameters,the SERCA2a protein expression was analyzed by Western Blot,and SERCA2a activity was carded out by modified Jones method.②The difference of proteome of hearts was detected by expression proteomics in between CHF+ SERCA2a group and CHF group at 30 days.③Electrophoretic separation and quantitation of cardiac myosin heavy chain(MHC) isoforrns were done at 30 days.RESULTS: ①In CHF group and CHF+GFP group, SERCA2a expression and activity were lower than those in sham-operation group.At 10 days after gene transfer.SERCA2a expression was increased by 80.7%and still lower than that of sham-operation group,and activity was also increased by 89.9%,but still lower than that of sham-operation group.And at 30 days,the SERCA2a protein expression and activity in the CHF+SERCA2a group reached to levels of sham-operation group. ②At day 10,LVSP and LV +dp/dtmax and LV-dp/dtmax were increased and LVEDP was decreased dramatically,but they still did not reach to the levels of sham-operation group.At day 30,the parameters were normalized back to control levels.There were no significant differences in each index between CHF+SERCA2a group and sham-operation group.③Overexpression of SERCA2a increased some energy metabolic enzymes of hearts at 30 days.④There were a downregulation of α-MHC and an upregulation of 3-MHC in failing hearts.Overexpression of SERCA2a increased the expression of α-MHC and decreased the expression of β-MHC to the levels of sham-operation group at 30 days.CONCLUSION: Adeno-associated viral gene transfer of SERCA2a can enhance SERCA2a functions, maintain calcium homeostasis,improve cardiac energy metabolism,and normalize the expression of MHC isoforms in CHF rats.As a result,the heart functions can be improved. So adeno-associated viral gene transfer of SERCA2a has good therapy effects on CHF rats in short and relatively long periods.