Objective: To evaluate the role of morphine preconditioning on necroptosis during myocardial ischemia-reperfusion (I/R) injury in the rats with heart failure. Methods: Clean-grade adult male Sprague-Dawley rats, weighing 200-230 g, were injected with 2 mg/kg doxorubicin via the tail vein once a week for 6 consecutive weeks to establish the chronic heart failure model.Thirty rats with chronic heart failure at the end of 8th week were divided into 3 groups (n=10 each) using a random number table method: sham operation group (group S), I/R group and morphine preconditioning group (group MPC). Myocardial I/R was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion in each group except group S. In group MPC, the rats were subjected to 3 cycles of 5-min infusion of 0.1 mg/kg morphine via the femoral vein at 5 min intervals before ischemia.The animals were sacrificed at the end of reperfusion, and the myocardial specimens were obtained for determination of the area at risk (AAR), infarct size (IS), expression of Fas mRNA (by quantitative real-time polymerase chain reaction) and expression of Fas, receptor-interacting protein 1 (RIP1) and RIP3 (by Western blot). The IS/AAR ratio was calculated. Results: Compared with group S, the IS and IS/AAR ratio were significantly increased at the end of reperfusion, and the expression of Fas protein and mRNA, RIP1 and RIP3 was up-regulated in group I/R (P<0.05). Compared with group I/R, the IS and IS/AAR ratio were significantly decreased at the end of reperfusion, and the expression of Fas protein and mRNA, RIP1 and RIP3 was down-regulated in group MPC (P<0.05). Conclusion The mechanism by which morphine preconditioning reduces myocardial I/R injury is related to inhibiting necroptosis in the rats with heart failure.

Objective To investigate the prevalence and drug resistance of clinical Klebsiella vari-icola ( K. variicola ) isolates and to illuminate the mechanism of drug resistance in carbapenem-resistant strains. Methods Clinical K. variicola isolates were identified with matrix-assisted laser desorption/ioniza-tion time-of-flight mass spectrometry ( MALDI-TOF MS ) . The antimicrobial susceptibility profile of these strains was determined using broth microdilution. Resistance genes carried by carbapenem-resistant K. vari-icola strains were detected by PCR with specific primers. Multilocus sequence typing ( MLST) was used for molecular typing. A pan-drug resistant strain which was isolated from cerebrospinal fluid sample was ana-lyzed with whole genome sequencing ( WGS) . Results Twenty-six isolates were identified as K. variicola by MALDI-TOF MS. Results of the antimicrobial susceptibility test showed that there were 15. 4％ (4/26) re-sistant to carbapenem and 11. 5％ (3/26) unsusceptible to tigecycline. These strains were highly suscepti-ble to amikacin and gentamicin, which accounted for 96. 2％ (25/26). As for the third-and fourth-genera-tion cephalosporins, the resistance rate was 23. 1％ (6/26). All of the four carbapenem-resistant isolates carried the resistance genes of blaIMP-4 , qnrA/B and blaTEM , and one of them was also positive for blaNDM-1 gene. The fosfomycin resistance gene, fosA, was detected in three of them. Molecular typing analysis indica-ted these isolates belonged to two sequence types ( ST) of ST357 ( three strains) and ST1737 ( one strain) . Two plasmids were obtained from the pan-drug resistant strain by WGS, including IncFⅡ/FIB( k) type plas-mid (160 kb) that was highly homologous to LMG 23571 plasmid (GenBank: CP013986. 1) and IncHⅠ1B/FIB type plasmid (260 kb) sharing high homology with pIMP4 LL34 (GenBank: CP025964. 1). Be-sides the resistance genes mentioned above, the two plasmids also carried a variety of other genes that media-ted the resistance to aminoglycosides (strB, strA, armA, aac3-Ⅱd, aadA2), macrolides (msrE, mphE), chloramphenicol (catA2), sulfonamides (sulⅠ) tigecycline (tetA variant) and trimethoprim (dfrA16). However, no virulence genes were detected. Conclusions In general, the resistance profile of K. variicola was similar to that of Klebsiella pneumoniae, but the differences were that carbapenem-resistant K. variicola strains mainly belonged to ST357 and the leading causes of resistance were carrying the genes encoding IMP-4 and NDM-1 metalβ-lactamases. WGC analysis revealed that the pan-drug resistant K. variicola strain carried multiple drug resistance genes without virulence determinants, which might be resulted from the evo-lution of drug resistance.

Objective To evaluate the role of μ opioid receptor in morphine preconditioning-induced reduction of myocardial ischemia-reperfusion (I/R) injury in rats with chronic heart failure.Methods Adult male Sprague-Dawley rats,weighing 170-230 g,in which chronic heart failure was induced by injecting doxorubicin via the tail vein,were studied.The rats were sacrificed and their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95％ O2-5％ CO2 at 37 ℃.Forty isolated rat hearts with I/R injury were randomly divided into 4 groups (n=10 each):group I/R,morphine preconditioning group (group MP),μ opioid receptor antagonist CTOP plus morphine preconditioning group (group CTOP+MP) and CTOP group.Myocardial I/R was induced by occlusion of the left coronary artery for 30 min followed by 120 min of reperfusion.In group MP,the hearts were perfused with K-H solution for 15 min,with K-H solution containing 1 μmol/L morphine for 5 min and with K-H solution for 5 min,3 cycles in total,and then the model of myocardial I/R was established.The hearts were perfused with K-H solution containing 1 μmol/L CTOP starting from 10 min before morphine preconditioning until 5 min of ischemia in group CTOP + MP.The hearts were perfused with K-H solution containing 1 μmol/L CTOP starting from 40 min before ischemia until 5 min of ischemia in group CTOP.The coronary effluent was collected at 15 min of equilibration (baseline) and 5 and 10 min of reperfusion to detect the activity of lactate dehydrogenase (LDH).Myocardial infarct size (IS) and the area at risk (AAR) were measured by 2,3,5-triphenyl-tetrazolium staining,and IS/AAR percentage was calculated.The expression of Bcl-2 and Bax mRNA was determined using uantitative real-time polymerase chain reaction,and the ratio of Bcl-2/Bax was calculated.Results Compared with group I/R,the IS and IS/AAR percentage were significantly decreased,the activity of LDH in coronary effluent was decreased,the expression of Bax mRNA was downregulated,the expression of Bcl-2 mRNA was up-regulated,and the Bcl-2/Bax ratio was increased in group MP (P＜0.05),and no significant change was found in the IS or IS/AAR percentage in CTOP and CTOP+ MP groups (P＞0.05).Compared with group MP,the IS and IS/AAR percentage were significantly increased,the activity of LDH in coronary effluent was increased,the expression of Bax mRNA was up-regulated,the expression of Bcl-2 mRNA was down-regulated,and the Bcl-2/Bax ratio was decreased in group CTOP+MP (P＜0.05).Conclusion The mechanism by which morphine preconditioning reduces myocardial I/R injury may be related to activating μ opioid receptors and thus maintaining the balance between Bcl2 and Bax gene expression in the rats with chronic heart failure.

Objective To compare the curative effect of open and minimally invasive treatment of acute closed achilles tendon rupture with 5 years followed-up study.Methods From September,2010 to January,2012,28 patients with acute closed Achilles tendon rupture in our hospital were followed up for 5 years.There were 21 males and 7 females.Minimally invasive percutaneous suture in 11 cases;open suture in the treatment of 17 cases.The patients were followed up at 6 months,1 year,2 years,3 years,4 years,and 5 years after AOFAS and ATRS score.Results The follow-up time ranged from 60 to 72 months.All incisions healed by first intention and no incision related complications occurred.2 groups of patients with ATRS score at 6 months after operation:the open group was 81.23±3.99,minimally invasive percutaneous group of 88.27±4.27,the difference between the two groups was statistically significantly.After 1 year,there was no significant difference in the scores between the two groups.Two groups of patients with AOFAS score at 6 months after operation:the open group was 69.00±6.23,minimally invasive percutaneous group of 79.27±4.83,the difference between the two groups was statistically significantly.At one year after operation,the open group was 85.53±3.38,and the minimally invasive group was more than 89.90±3.38.The difference between the two groups was statistically significantly.After 2 years,there was no significant difference in the scores between the two groups.Conclusion There is no significant difference between the 2 years after surgery in the treatment of acute closed Achilles tendon rupture or open surger.

With the continuous application of high-technology in the clinical practice,the privacy of patient is infringed or leaked during the process of imaging medical examination and teaching or because of improper management of hospital.The main reasons include that relative laws and regulations are inadequate,patients don't understand the scope of privacy rights and medical technicians lack knowledge of medical ethics.From the angle of medical ethics,this paper further discussed the protection of patients' privacy in imaging medical examination,so as to effectively reduce medical disputes in practical work and build a good doctor-patient relationship.

Aim To investigate the effects of lentivirus mediated nerve growth factor ( NGF) gene silencing on pheochromocytoma cells ( PC12 ) and the possible mechanisms .Methods The NGF shRNA expression vector was constructed .PC12 cells were randomly divi-ded into five groups (n=3 each) as follows: negative control group ( NC ) , control lentivirus group ( LV CON) , lentivirus NGF shRNA1 group ( LV shNGF1 ) , lentivirus NGF shRNA2 group(LV shNGF2), lentivir-us NGF shRNA3 group(LV shNGF3).The cells in NC group were cultured in DMEM/HG and polybrene me-dium, while others were cultured in DMEM/HG, poly-brene and corresponding lentivirus medium .After the treatment, the infection efficiency was determined by fluorescent microscope .Relative expression of NGF , extracellular signal-regulated kinase ( ERK1/2 ) and p-ERK1/2 were assessed by Western blot .The expres-sion of NGF mRNA was analyzed by quantitative re-verse transcription polymerase chain reaction ( qRT-PCR) .The differentiation degree was valued according to the length of neuritis and max diameter of cells .The cell viability was detected by CCK-8.Results The in-fection efficiency in PC12 cells reached over 90%. Compared with NC group , the relative expression of NGF mRNA and NGF protein was significantly down-regulated ( P<0.05 ) .There was no difference in the expression of ERK1/2 protein and cell viability .The expression of p-ERK1/2 protein was markedly down-regulated in LV shNGF3 group ( P<0.01 ) .The cells morphology was changed , and the length of neuritis and max diameter of cells were strained in LV shNGF 3 group than those in NC group ( P<0.01 ) .Conclusion Lentivirus-mediated NGF gene silencing inhibits the differentiation of PC12 cells through suppressing the activation of ERK1/2.

Purpose To discuss the value of MRI in the diagnosis of placenta implantation abnormality, and to explore preliminarily the relationship between MRI signs and types of placenta implantation abnormality. Materials and Methods The clinical preoperative data and postnatal pathological findings of 54 women at high risk of placenta accreta were collected. All the patients undertook the conventional pelvic MRI examination. The scanning sequences mainly included: sagittal, coronal and axial T2-weighted imaging-turbo spin echo, balance fast field echo. The MRI images were observed and the areas which showed low signal in all the three directions on T2WI were measured. Then the correlation between the areas of low signal on T2WI in placental and the types of placenta implantation abnormality was analyzed. Results The incidence of placenta implantation abnormality was 64.8% in our research (35/54). The main MRI signs were low signal on T2WI (68.5%, 37/54) and heterogeneous signal in placenta (57.4%, 31/54); the main sign of placenta percreta was tenting bladder (75.0%, 6/8). The types of placenta implantation abnormality were positively correlated with the areas of low signal on T2WI (r=0.454, P<0.05). Conclusion Pregnant women at risk of placenta accreta should be evaluated with imaging examinations, particularly with MRI scanning, to improve disease detection rate. The typical indirect signs of placenta implantation abnormality are low signal on T2WI and heterogeneous signal in placenta. The larger size of low signal area on T2WI in placenta, the deeper implantation of placenta.

Objective To investigate the effects of early physiotherapy in combination with atorvastatin on the levels of serum brain-derived neurotrophic factor (BDNF) and neurological function in patients with acute ischemic stroke.Methods Fifty patients with acute ischemic stroke were randomly divided into either an atorvastatin group (monotherapy group,n =25) or a early physiotherapy + atorvastatin group (combination treatment group,n =25).All patients received the prescribed drugs according to the diagnosis and treatment guidelines for ischemic stroke.The monotherapy group added atorvastatin calcium (20 mg,1 tablet every night orally).On the basis of the monotherapy group,the combination treatment group also conducted early physical therapy.At 2 and 6 weeks before and after treatment,a double-antboody sandwich enzyme-linked immunosorbent assay was used to detect the serum BDNF levels.The National Institutes of Health Stroke Scale (NIHSS) was used to evaluate the degree of neurological deficit.Barthel index (BI) was used to evaluate the activities of daily living.The modified Rankin scale (mRS) was used to assess the degree of disability.Results There was no significant difference in demographics and baseline data between the monotherapy group and the combination treatment group.The scores of NIHSS,BI,and mRS in both groups after treatment were significantly better than those before treatment (all P ＜ 0.001).There were no difference in the scores of NIHSS,BI and mRS at 2 weeks before and after treatment,but at 6 weeks after treatment,the scores of NIHSS (2.40 ± 1.38 vs.3.36 ± 1.73; P =0.035) and mRS (1.40 ± 0.87 vs.1.96 ±0.94; P =0.047) of the combination treatment group were significantly lower than those of the monotherapy group,and the BI scores (92.60 ±7.50 vs.85.20 ± 11.68; P=0.011) were significantly higher than those of the monotherapy group.After treatment,the serum BDNF levels were increased significantly in both groups.There were significant differences among all the time points (all P＜0.001).At 2 weeks after treatment,the serum BDNF levels (3.07 ±0.93 ng/ml vs.2.45 ±0.76 ng/ml; t =2.559,P =0.014) and at 6 weeks after treatment,those (2.90 ± 0.93 ng/ml vs.2.31 ± 0.77 ng/ml; t =2.433,P =0.019) in the combination treatment group were significantly higher than those in the monotherapy group.Spearman correlation analysis showed that the serum BDNF levels were significantly negatively correlated with the scores of NIHSS (r =-0.738,P ＜ 0.001) and mRS (r =-0.654,P ＜ 0.001),but they were significantly positively correlated with the BI scores (r =0.716,P ＜ 0.001).No serious adverse reaction occurred in both groups.Conclusions Early physiotherapy in combination with atorvastatin for the treatment of acute ischemic stroke can more effectively promote the recovery of neurological function,and its mechanism may be associated with the increased serum BDNF levels.

OBJECTIVEAttempting to isolate and cultivate the microcytin-RR-producing cyanobacteria from natural blooms as well as to further investigate some characteristics of their growth and metabolite toxicity.

METHODSCapillary-pipette method was used to isolate wild Microcystis strains collected from eutrophicated lakes. The isolated strains were cultured in BG11 media at (25 ± 1) °C, under 2 000 lx illumination of fluorescent light with a light-dark rhythm of 12-12 h. The growth curve was observed by measuring optical density of culture suspension, toxin-related genes and the metabolite toxins were identified separately by PCR and HPLC, and its acute toxicity was carried out by orally administered toxins to Kunming (KM) mice.

RESULTSOne of five toxigenic strains from 198 collected samples was confirmed to be a MC-RR producing blue-green alga by existing two specific toxin-synthesized enzyme genes and showing specific chromatographic peak of the toxin compared with standard MC-RR through both PCR and HPLC methods. The toxic strain was classified as Microcystin aeruginosa by morphologic and phylogenetic tree analysis. The growth length of the strain lasted nearly 81 days with 55-60 days' exponential phase and the maximal concentration of 5.52 × 10⁷ cell/ml. The LD50 of the MC-RR to the KM mice ranged from 10.75 mg/kg to 13.45 mg/kg of body weight. As a result of the acute toxicity, the enzymatic indexes in serum such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) were significantly higher than those in the control group. The levels of ALT, AST, ALP and LDH in the treated group at 45 min were (157.08 ± 20.38), (333.00 ± 68.53), (392.70 ± 89.59) and (1 071.13 ± 160.22) U/L respectively, and at 4 h were (514.68 ± 156.87), (593.15 ± 40.41), (618.55 ± 208.76) and (2 281.72 ± 866.67) U/L respectively, and meanwhile the values of ALT, AST, ALP and LDH in the control group were (40.30 ± 4.89), (142.70 ± 26.59), (56.90 ± 11.89) and (509.50 ± 94.75) U/L separately (t values at 45 min were -11.20, -5.77, -7.38, -6.60 respectively, and at 4 h were -6.04, -20.21, -5.35, -4.07 respectively, P values were all <0.01). The liver coefficient in the treated group at 45 min and 4 h were 6.855 ± 0.225 and 8.409 ± 0.276, significantly higher than that (5.784 ± 0.286) in the control group (t values were -3.96 and -12.22, P values were both <0.01). The histopathological changes of liver were hyperemia obviously.

CONCLUSIONIsolated from the bloom waters, a strain of Microcystis aeruginosa is obtained with characteristics of longer growth duration, positive microcystin synthetase genes, and dominant production of MC-RR. The LD50 of the extracted MC-RR administered by oral route to mice is (12.10 ± 1.35) mg/kg of body weight, and liver is the target organ of MC-RR. The existence and potential risk of MC-RR in China cannot be ignored.

Animals ; China ; Chromatography, High Pressure Liquid ; Cyanobacteria ; Hyperemia ; Lakes ; Liver ; Mice ; Microcystins ; Microcystis ; Phylogeny

Brain-derived neurotrophic factor (BDNF) is one of the most prevalent growth factors in the central nervous system (CNS).In the development and maturation processes of the nervous system,BDNF plays an important role in maintaining neuronal function,promoting neuronal regeneration after injury,and preventing neuronal degeneration,etc.At present,many researchers are being dedicated to the research of BDNF for treatment of brain ischemia and have achieved some progress.This article reviews the molecular biological characteristics and biological function of BDNF,roles and mechanisms in cerebral ischemia,and the possibility as an intervention target of cerebral ischemia.