Objective To investigate molecular typing and drug resistance patterns of 98 Klebsiella pneumoniae (K.pneumoniae) isolated from type 2 diabetes patients complicated with maxillofacial infection,to research the virulence and resistance mechanisms.Methods The study was a prospective study that adopted the method of continuous sampling from fixed location,from March 2010 to October 2012.The maxillofacial surgery patients diagnosed with type 2 diabetes complicated with maxillofacial infection were chosen in 7 hospitals in Zhengzhou as the research object,and a total of 431 pus sample were collected continuously,in which 98 strains K.pneumoniae were isolated and identified.The Kirby-Bauer disk diffusion test was conducted in 98 strains to determine the resistance to 19 antibacterial agents.K.pneumoniae chromosomal DNA were digested by restriction endonuclease Xba Ⅰ and analyzed by pulsed-field gel electrophoresis (PFGE).PFGE patterns of K.pneumoniae strains were analyzed using Fingerprinting software.The relationship between the molecular types and resistance phenotype was observed.The extended spectrum β-1actamase-producing K.pneumoniae were screened out by the double disc synergy test (DDST)Polymerase chain reaction (PCR) was used to detect resistant genotypes,serotype and virulence genes.The purified PCR products of resistant genes were cloned and sequenced.Hypermucoviscosity phenotype of all strains were determined by string test.Results Much severer drug-resistance for K.pneumoniae was identified and the result of extended-spectrum beta-lactamase producing rate was 57.1 ％.Ninety-eight strains were dispatched into 13 groups by PFGE.No dominant bands and specific extended-spectrum beta-lactamase DNA bands were found.The results of PCR showed that among the 56 strains of extended spectrum β-lactamase-producing K.pneumoniae,40 were positive for blaSHV (accounting for 71.4％),28 positive for blaTEM (accounting for 50.5％),21 positive for blaCTX-M (accounting tor 37.5％).The sequencing results were as follows:TEM-1,CTX-M-3 and a variety of SHV.Serotype K1,K2,K3,K5,K20,K54 and K57 and 3 kinds of virulence genes were detected,but not in strong toxicity-based.Hypermucoviscosity positive rate was 31.6％ (31/98).Conclusion Much severer drug resistance of K.pneumoniae in this study was identified and resistant mechanism was complex,in which strong toxicity serotype and virulence geues exist,which need more attention from clinical.

Objective: To study the effects of total flavonoids of Astragalus on apoptosis of bovine retinal capillary pericytes(BRPs) under high glucose.Methods: The third generation of nearly symphysic bovine retinal vessel pericytes cultivated in vitro were divided into normal control group,high glucose group,and Astragalus total flavonoids groups(0.25,0.5,1.0,2.0 mg/ml) at random.After being incubated for 6 days,apoptosis of BRPs were detected by TUNEL method.TBA method was used to detect the contents of MDA.Xanthine oxidase method was used to detect the SOD activities.Results: Compared with high glucose group,the apoptosis,MDA contents,SOD activity,MDA content/SOD activity of BRPs reduced markedly in total flavonoids of Astragalus groups(0.5,1.0,2.0mg/ml)(P

Objective:To analyze the categories, scoring methods and effects of dietary patterns in heart failure patients, systematically.Methods:The databases such as Medline, Embase, and Cochrane were searched to obtain articles on dietary patterns of heart failure patients up to December 31, 2019. Arksey and O′Malley's scoping review method was applied to analyze the articles.Results:A total of 1 500 studies were identified, 21 of them met the inclusion criteria. These studies mainly studied Mediterranean diet(11 articles), Dietary Approaches to Stop Hypertension Diet(7 articles), Palaeolithic diet(1 article), low fat diet(1 article), High-protein Diet(1 article).Conclusions:The Mediterranean and DASH dietary patterns have a positive protective effect on heart failure disease prevention and treatment, but more randomized controlled trials are needed to improve the reliability of the conclusions.

Objective To investigate the prevalence of nutritional risk,undemutrition,overweight/obesi-ty,and apphcation of nutrition support in departments of neurology in 3 metropolitan hospitals in Beijing. Methods Adult patients in the departments of neurology from 3 metropolitan hospitals in Beijing were consecutively en-rolled from March 2005 to March 2006.Nutritional Risk Screening 2002(NRS2002)score≥3 was defined a8 nu-tritian risk,while body mass index(BMI)<18.5 ks/m2 as undemutrition.NRS2002 was performed on the next morning of admission and nutritional support evaluation was performed on the 14th day of admission or discharge day.The relationship between nutrition risk and nutrition support was analyzed.Results A total of 753 patients were enrolled.NRS2002 scoring results were obtained in 461 patients.The prevalence of undemutrition was 4.2%and nutritional risk Was 21.2%.Among them 292 cases in undemutrition category were estimated by semm albu-min<30g/L without accurate BML If the cases without accurate BMl were excluded,the prevalence of undemutri-tion was 5.4%and nutritional risk Was 10.8%.Twenty-three patients(14.4%)with NRS2002≥3 received nutrition support.Thirteen patients(2.2%)with NRS2002<3 received nutrition support.The average PN:EN ratio Wag 1:2.Conclusions A large proportion of inpatients were at nutritional risk or undemutrition in the de-partments of neurology in metropolitan hospitals in Beijing.The application of nutrition support Wag somehow inap-propriate in these patients.Evidence-based guidelines are required to improve this situation

Objective To study the effects of constant and intermittent high glucose on the apoptosis of cultured bovine retinal capillary pericytes (BRPs), and to investigate the role of mitochondrial apoptotic pathway in the apoptosis of BRPs. Methods After being cultured under glucose with different concentrations tot 6 days, the change of uhrastructure of BRPs was observed under electronmieroscope, the apoptosis of pericytes was detected by TUNEL method, the mitochondrial inner transmembrane potential (△Ψm) was detected by laser scanning confocal microscopy, the change of cytochrome c (cyt-c) was assayed by spectrophotometer and the expression of apoptotic genes was detected by immunohistochemical method and RT-PCR. Results (1) BRPs showed typical changes of apoptosis in constant and intermittent high glucose concentrations. The apoptosis induced by constant high glucose concentration was more obvious than that by intermittent high glucose. (2) Constant and intermittent high glucose concentrations obviously decreased △Ψm compared with control group. The △Ψm of BRPs was correlated negatively with the apoptotie rate of BRPs (r = - 0.89, P < 0.01) ; (3) Constant and intermittent high glucose concentrations increased the release of cyt-c from mitochondria to cytoplasm, and the concentration of cyt-c in the cytoplasm was correlated positively with the apoptotic rate of BRPs (P < 0.01) ; (4) Constant and intermittent high glucose concentrations increased the expression of proapoptotie gene Bax and decreased the expression of prosurvival gene Bcl-2, resulting in increased Bax/Bcl-2 ratio. Bax/Bcl-2 ratio was negative correlatied with the △Ψm of BRPs, and positively eorrclaticd with the concentration of cyt-c in cytoplasm and apoptotic rate (both P < 0.01).Conclusion Constant and intermittent high glucose concentrations could decrease △Ψm, increase the release of cyt-c and induce the apoptosis of BRPs, the effects being stronger with constant high glucose concentration. The mitochondrial apoptotic pathway plays an important role in the apoptosis of BRPs, in which Bax and Bcl-2 are involved.

AIM:To study the effects of noninvasive delayed limb ischemia preconditioning ( NDLIP) on ani-mal cardiac function , myocardial morphology and myocardial apoptosis after myocardial infarction ( MI ) .METHODS:Healthy SD male rats [n=45, weighing (250 ±10) g] were randomly divided into 3 groups:MI group:the animal model of MI was established by surgical ligation of left anterior descending artery ( LAD) after 2 weeks;NDLIP group:after the success of the MI animal model , NDLIP was carried out every other day until the 4th, 6th and 8th weeks;sham group:as the negative control group , the animals were taken heart LAD threading but no ligation .All rats were fed conventionally .At the end of the 4th, 6th and 8th weeks, all rats were made ventricular intubation , and then the hemodynamic parameters were recorded .The blood samples were withdrawn from the abdominal aorta and the serum was separated via centrifugation . The serum contents of Bcl-2 and Bax were measured by ELISA .Left ventricular anterior wall was homogenized .The mito-chondrial respiratory chain complexes Ⅰ,Ⅱ,Ⅲand Ⅳin the myocardial tissues were detected by ELISA .RESULTS:At the end of the 4th, 6th and 8th weeks, compared with MI group, left ventricular systolic pressure in NDLIP group was significantly increased , while left ventricular end-diastolic pressure in NDLIP group was significantly decreased ( both P<0.05).Mitochondrial respiratory chain complexesⅠ, Ⅱ, Ⅲ and Ⅳ in NDLIP group were significantly increased (P<0.05).The serum level of Bcl-2 in NDLIP group was significantly increased and Bax level was reduced remarkably (both P<0.01) .CONCLUSION:NDLIP improves the hemodynamic indexes , promotes the mitochondrial respiratory function and inhibits cell apoptosis , thus improving the prognosis of MI .

Aim To study the preventative effects of noninvasive delayed limb ischemic preconditioning ( NDLIP) on sudden cardiac death in rats with myocar-dial infarction. Methods Thirty healthy SD male rats weighting ( 250 ± 10 ) g were randomly divided into 3 groups:① myocardial infarction ( MI ) group: animal model of MI was established by making surgical ligation of animal LAD. ② MI plus NDLIP group: after the success of the animal model of MI, NDLIP was carried out every other day until 4th week. ③Sham group:as the negative control group, animals were taken heart LAD threading but no ligation. All rats were fed con-ventionally. At the end of 4 weeks, three groups of rats were administered with metaraminol ( 0. 2 mg · min-1 ) . ECG, drug cumulant of sudden death and death onset time were recorded. After sudden death, blood samples were withdrawn from abdominal aorta and serum was separated via centrifugation. ELISA method was used to measure serum caspase-3 , HSP70 and SOD concentration. Results While metaraminol led animal cardiac sudden death, the rats heart rate ( HR) kept declining with the increase of dosage of metaraminol during the administration period. Rat HR of MI+NDLIP group [ ( 479 ± 8 ) vs ( 416 ± 19 ) beat ·min-1 , ( 446 ± 32 ) vs ( 370 ± 20 ) beat · min-1 , (376 ± 53) vs (305 ± 29) beat·min-1, (307 ± 63) vs (244 ± 33) beat·min-1, (283 ± 45) vs (121 ± 35 ) beat · min-1 , P <0. 01 ] was markedly higher than that of MI group at 0 , 5 , 10 , 30 , 50 min before death. Compared with MI group, drugs cumulant to sudden death and death onset time of MI + NDLIP group [ ( 14. 58 ± 3. 03 ) vs ( 10. 76 ± 2. 73 ) mg, (72. 9 ± 15. 2 ) vs ( 53. 8 ± 13. 6 ) min, P <0. 01 ] were significantly increased. Compared with MI group, serum caspase-3 content of MI+NDLIP group was sig-nificantly reduced [ ( 2. 01 ± 0. 52 ) vs ( 2. 34 ± 0. 38 )μg·L-1 , P<0. 01 ]; HSP70 levels were remarkably increased [ ( 3. 01 ± 0. 58 ) vs ( 2. 70 ± 0. 43 ) μg · L-1 , P <0. 05 ]; SOD levels were significantly im-proved [(1. 99 ± 0. 65) vs (1. 70 ± 0. 58) mg·L-1, P<0. 01 ] . Conclusion NDLIP can prevent sudden cardiac death after myocardial infarction in rats, which may be mediated by reducing the myocardial cell apop-tosis, increasing protective protein expression and en-hancing antioxidant capacity.

Objective:To explore the protective effect of miR-346 up-regulated by crocin on myocardial ischemia reperfusion injury in rats.Methods:The rat model of myocardial ischemia reperfusion was constructed by open-chest ligation of the left anterior descending coronary artery followed by reperfusion. Fifty male SD rats were randomly divided into the sham operation group, model group, crocin group, miR-346 negative control (miR-NC) group, and miR-346 inhibitor group with 10 rats in each group. Rats in the sham operation group were only received thoracotomy without ligation. Rats in the miR-NC group and miR-346 inhibitor group were injected with miR-NC or miR-346 inhibitor through the tail vein 48 h prior to ligation of the left anterior descending coronary artery ligation. The crocetin reagent for gastric lavage was prepared by dissolving 8 mg of crocetin in 100 ml of physiological saline. Rats in the crocetin group, miR-NC group, and miR-346 inhibitor group were gavaged with crocetin reagent at 80 mg/kg. Rats in the sham operation group and model group were gavaged with saline at 5 ml/kg. The crocetin reagent and saline were gavaged once a day for 15 days. Serum levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) were detected by enzyme-linked immunosorbent assay (ELISA). The miR-346 expression level was detected by real-time fluorescence quantitative PCR (qRT-PCR). The pathological changes in cardiac muscle tissues were detected by HE staining. Cardiomyocyte apoptosis rate was detected by TUNEL staining. The expression levels of apoptosis-related proteins were detected by Western Blot.Results:Compared with the sham operation group, the serum levels of CK-MB and LDH, pathological scores, cardiomyocyte apoptosis rate and expression of cleaved cysteinyl aspartate specific proteinase-3 (cleaved Caspase-3) and B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax) were increased in the model group (all P < 0.05), and the miR-346 expression level and Bcl-2 levels in myocardial tissues were decreased (all P < 0.05). Compared with the model group, serum CK-MB and LDH levels, pathological scores, cardiomyocyte apoptosis rate, and cleaved Caspase-3 and Bax expression levels were decreased in the crocin group (all P < 0.05), and miR-346 expression level and Bcl-2 levels in the myocardial tissues were increased (all P < 0.05). Compared with the miR-NC group, the serum levels of CK-MB, LDH, pathological scores, cardiomyocyte apoptosis rate, and cleaved Caspase-3 and Bax expression levels were decreased in the miR-346 inhibitor group (all P < 0.05), and miR-346 expression level and Bcl-2 level were increased in myocardial tissues (all P < 0.05). Conclusions:Crocin can reduce myocardial tissue injury and attenuate cardiomyocyte apoptosis in myocardial ischemia-reperfusion rats by up-regulating miR-346.

This study performed to determine whether OX40L overexpressed by recombinant rabies virus(LBNSE-OX40L)can enhance the innate immune response through activation of dendritic cells and thus activate early antibody production.Bone marrow dendritic cells(BMDCs)were extracted from the femur of Balb/c mice and cultured for 6 days,and the cultured BMDCs were infected with the parental virus LBNSE and the recombinant virus LBNSE-OX40L with the multiplicity of infections(MOI)=1.The effect of each virus on the maturation of BMDCs was analyzed by flow cytometry;ELISA was used to detect the expression of innate immunity-related cytokines such as interferon-α(IFN-α)and interleukin-12p40(IL-12p40)in the supernatants of the infected BMDCs.For in vivo study,Balb/c female mice were injected intramuscularly with 106 FFU of parental virus LBNSE and recombinant virus LBNSE-OX40L in both hind limbs,and the inguinal lymph nodes of mice were collected on day 6 after immunization,and the proportion of mature DCs was detected by flow cytometry.The serum was collected on day 6 after immunization,and the content of virus-neutralizing antibody(VNA)was detected by antiviral neutralizing antibody titration.Mouse serum was collected on day 6 after immunization,and virus neutralizing antibody content was measured by titration of antiviral neutralizing antibody,while IgG antibody in mouse serum was detected by ELISA.IgM antibody subclasses were detected by ELISA on days 2,4,and 6 after immunization.Compared with the parental virus LBNSE,the recombinant virus LBNSE-OX40L was able to activate more BMDCs in vitro and produce significantly higher levels of IFN-α and IL-12p40.Furthermore,the recombinant virus LBNSE-OX40L stimulated the maturation and differentiation of the DCs in vivo,which led to the rapid production of high levels of VNA and RABV-specific IgG and IgM antibodies.Taken together,LBNSE-OX40L activates dendritic cells to promote the body's innate immune response,and in turn enhances early antibody production,thus can be an early effective rabies vaccine candidate.

Objective:To observe the effect of Chinese herbal compound Siwu decoction (SW) on radiation injury of zebra fish blood system, and to clarify whether zebra fish is a suitable model for this study. Methods:Totally 72 of 4-month-old adult male zebra fish ( Danio Rerio) with body weight ranging from 0.14 to 0.20 g were used in all the experiments, where 36 zebra fish were taken to observe the hemodynamic changes after radiation, and the other 36 zebra fish were used to study the intervention effect of Siwu decoction on the blood injury after radiation. Zebra fish was irradiated with 20 Gy of 60Co gamma-rays at a dose rate of 97.33 cGy/min. For the hemodynamic study, the peripheral blood and whole kidney marrow cells (WKMCs)of irradiated zebra fish and non irradiated control were collected on days 7, 14 and 30 after irradiation, respectively, and the cell number was detected by flow cytometry. For the Siwu decoction treatment, 36 zebra fish were randomly divided into non-irradiation group, irradiation group, SW2 000 group ( Siwu decoction was diluted by 2 000-fold) and SW5 000 group ( Siwu decoction was diluted by 5 000-fold). Siwu decoction was given on the second day after irradiation. Then the peripheral blood and WKM cells of zebra fish were collected at 7 d after irradiation for further analysis. Results:After irradiation, the dynamic of blood development of zebra fish were obviously changed with the most blood cell ablation at 7 d post-irradiation. The total number of peripheral blood cells and WKM cells decreased by 26% and 52%( t=4.535, 28.987, P<0.05), the number of myeloid monocytes, lymphocytes, and red blood cells in the WKM decreased by 46%, 79% and 33%( t=18.457, 66.900, 9.872, P<0.05), and the number of precursor cells decreased by 49% ( P>0.05). The numbers of these blood cells were partly recovered at 14 d after irradiation and returned to the normal level at 30 d after irradiation. In comparison with irradiation alone group, the numbers of WKM cells, myeloid monocytes, progenitor cells, lymphocytes and red blood cells increased by 57%, 125%, 81% and 35%, respectively ( t=12.128, 21.594, 15.473, 4.594, P<0.05), and peripheral red cells did not increase significantly in the SW5 000 group after irradiation. However, SW2 000 had no significant influence on radiation injury of zebra fish blood cells ( P>0.05). Conclusions:Radiation injury of the blood system of zebra fish could be recovered at a month after irradiation, and this radiation damage could be partly intervened by Siwu decoction at a suitable concentration, which provides a reference for the study of blood system regulation of traditional Chinese medicine.