Objective To understand the clinical distribution characteristics and drug-resistance genotypes of carbapenem-resistant Pseudomonas aeruginosa(CRPA)in a tertiary hospital in Shanghai,so as to guide the rational use of antibiotics,reduce bacterial resistance and control nosocomial infections.Methods A total of 94 consecutive and non-redundant CRPA strains isolated from clinical specimens were collected in The First Affiliated Hospital of Naval Medical University between Jan.1,and Dec.31,2019.The distribution of CRPA across departments and sample sources were analyzed.Antimicrobial susceptibility testing against 13 antibiotics was determined using the VITEK 2 Compact automated microbiology analyzer and the Kirby-Bauer disk diffusion method.Polymerase chain reaction(PCR)was employed to detect genes encoding extended-spectrum β-lactamase resistance gene,carbapenemase resistance gene,and porin resistance gene.Homology analysis of selected isolates was performed using kSNP3,a k-mer-based software,for single nucleotide polymorphism(SNP)analysis.An evolutionary tree was constructed to assess genetic relationships.Additionally,multilocus sequence typing(MLST)was performed using the Pseudomonas aeruginosa typing scheme from the PubMLST database.Results Among the 94 CRPA isolates,high resistance rates were observed for carbapenems,including imipenem(92.6%,87 strains)and meropenem(87.2%,82 strains).In contrast,low resistance rates were detected for aminoglycosides,such as amikacin(10.6%,10 strains),gentamicin(20.2%,19 strains),and tobramycin(20.2%,19 strains).The top 3 departments in terms of isolate distribution were the Emergency Intensive Care Unit(9.6%,9 strains),Department of Cerebrovascular Surgery(8.5%,8 strains),and Department of Respiratory Medicine(8.5%,8 strains).PCR analysis of 94 CRPA strains detected outer membrane protein D2(OprD2)gene deletion in 47(50.0%)strains,13(13.8%)strains were positive for blaKPC,4(4.3%)strains for blaVIM,2(2.1%)strains for blaIMP,1(1.1%)strain for blaNDM,12(12.8%)strains for blaTEM,4(4.3%)strains for blaPER,and 2(2.1%)strains for blaGES,while blaOXA-48,blaBIC,blaSIM,blaVEB,and blaSHV were not detected.MLST identified 36 different sequence types(STs),with ST463 and ST274 being the most common,and 2 new ST(ST4023 and ST4024)were identified for the first time.Conclusion CRPA strains carry multiple resistance genes and exhibit concurrent resistance to several commonly used clinical antibiotics.The resistance is primarily associated with the presence of blaKPC,blaVIM and blaTEM genes and the deletion of OprD2 gene.Clinical monitoring of CRPA should be strengthened,and rational use of antimicrobial agents is essential to control its spread within the hospital.

The study adopted muscle injection of pVAX1-SP-GHRH(1-44)expression plasmid and electrostimulation to determine its effects on mouse growth and sow production performance.One hundred and fifty four-week-old C57 BL/6 female mice were randomly divided into 6 groups of 5 replicates each.Muscle single-injection followed by electrostimulation was performed.The con-trol group received an empty plasmid injection(80 μg/kg),while the treatment groups received pVAX1-SP-GHRH(1-44)plasmid injections(20,40,80,120,160 μg/kg).Twenty healthy preg-nant sows were randomly divided into 2 groups,each with 10 sows.Electrostimulation treatment was applied to the semimembranosus muscle of the pregnant sows after a single injection.The con-trol group received physiological saline injection,while the plasmid group received a 2 mg pVAX1-SP-GHRH(1-44)expression plasmid injection.Mouse weight,feed intake,and serum GHRH and IGF-1 levels were measured at days 0,7,14,21,and 28 after injection.Pregnant sows were bled via the tail vein at days 0,14,28,and 42 after injection,and their serum was separated to measure serum GHRH and IGF-1 levels.The birth weight,placental weight,number of piglets born,number of healthy piglets,number of weak piglets,number of deformed piglets,number of stillborn piglets,and number of mummified piglets were recorded at day 14.The mouse study re-sults showed that muscle injection of pVAX1-SP-GHRH(1-44)plasmid followed by electrostim-ulation could promote mouse feeding and increase weight gain(P＜0.05),significantly increase mouse serum GHRH and IGF-1 levels(P＜0.05),and maintain its effects until day 21.The results of the pregnant sow study showed that the average birth weight of the piglets in the plasmid group was significantly increased(P＜0.01),and the placenta weight was significantly increased(P＜0.05).The serum GHRH and IGF-1 concentrations in the plasmid group sows were significantly increased(P＜0.01).The study results showed that muscle injection of pVAX1-SP-GHRH(1-44)expression plasmid followed by electrostimulation could promote mouse feeding and increase weight gain,and also significantly improve the average birth weight and placental weight of the piglets in pregnant sows.

Objective To study the concentration distribution of acetone in fasting exhaled breath in diabetic patients and healthy subjects,to explore the effect of individual indexes on the concentration of acetone in fasting exhaled breath,and to study the role of individual indexes of fasting exhaled breath acetone in diabetes screening.Methods The acetone concentration measurements of fasting exhaled breath were performed on 265 healthy subjects,39 patients with type 1 diabetes (T1D),and 300 patients with type 2 diabetes (T2D) using real-time online respiratory acetone analyzer based on cavity ring-down spectroscopy (CRDS).SPSS 19.0 software was used to eliminate outliers,and relevant statistical analysis was carried out with the corresponding gender,age,height,body mass,body mass index (BMI) and blood glucose concentration (BGL).The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of fasting breath acetone concentration for diabetes diagnosing.Results The mean fasting breath acetone concentration in T1D patients was (2.24±1.43)×10-6 was significantly higher than (1.43±0.55)×10-6 in healthy subjects and (1.41±0.73)×10-6 in T2D patients,and the differences were statistically significant (all P＜0.05).The average fasting breath acetone concentration in male diabetic patients was higher than that in female patients.The mean fasting breath acetone concentration was positively correlated with age (R=0.31,P＜0.01) in healthy subjects,was positively correlated with BMI (R=0.33,P＜0.05) in T1D patients,and was positively correlated with height (R=0.18,P＜0.01) in T2D patients.The area under the ROC curve for the diagnosis of T1D by fasting breath acetone concentration was 0.853 with a sensitivity of 71.9％ and specificity of 87.4％ (P＜0.01),and for the diagnosis of T2D was 0.528 with a sensitivity of 54.1％ and specificity of 55.0％ (P＞0.05).Conclusions The detection of fasting breath acetone concentration is meaningful for T1D diagnosing,but has a low accuracy for T2D diagnosing (no statistically significant).

Objective To investigate the neuroprotective effect and mechanism of liraglutide on diabetic rats. Methods 24 healthy male SPF Goto-Kakizaki (GK) rats with random blood glucose greater than 11.1 mmol/L were selected as the experimental group, and randomly divided into diabetes mellitus group ( n＝12) and liraglutide group (n＝12). Ten healthy male SPF Wistar rats with the same age and weight as GK rats were selected as normal control group. After adaptively feeded for 2 weeks, the liraglutide group was given liraglutide (400 μg·kg－1·d－1, subcutaneous injection), while the control group and diabetes mellitus group were given the same volume of saline, and continued to be administered for 8 weeks. After 10 weeks, data and biochemical indicators were recorded. Effects of liraglutide on learning and memory in diabetes mellitus rats were detected by Morris water maze test. HE staining observed the hippocampal neurons morphology. Western blotting method detected the expression of p- IκB kinase (IKK) β, p-NF-κB, NF-κB, Klotho, and PRX2 in hippocampus. Results Morris water maze test showed that liraglutide can improve the spatial learning and memory ability of diabetes mellitus rats. HE staining showed that liraglutide significantly reduced the pathological damage of hippocampal neurons of diabetes mellitus rats. Western blotting showed that liraglutide inhibited NF-κB signaling pathway in hippocampus of diabetes mellitus rats. The expression of Klotho protein in hippocampus of diabetes mellitus group was significantly lower than that of control group, while the expression of PRX2 protein was higher than control group (t＝8.298,－7.398,all P<0.01). The expression of Klotho and PRX2 protein in hippocampus of liraglutide group were higher than diabetes mellitus group (t＝－13.059, 14.113, all P<0.01). The expression of Klotho protein of liraglutide group was similar to that of control group ( t ＝ －1. 137, P>0. 05 ). The expression of PRX2 protein was significantly higher than control group (t＝－28.055, P<0.01). Conclusions Liraglutide may enhance the expression of antioxidant stress protein including Klotho and PRX2, by inhibiting NF-κB signaling pathway in hippocampus of diabetes mellitus rats, reduced oxidative stress and improved the injury of hippocampal neuronal in diabetes mellitus rats, which seems to play a neuroprotective effect, to prevent and delay the occurrence of diabetic encephalopathy.

Objective To investigate the changes of C- reactive protein( CRP ) and homocysteine ( Hcy)in the type 2 diabetes with depression,and its clinical significance and potential mechanism. Methods One hundred and twenty-four cases with type 2 diabetes were divided into the depression group(63 cases)and non-depression group( 61 cases ) according to the Self-Rating Depression Scale and verified by Self-Rating Anxiety Scale. The information including age,sex,education degree,body mass index,course of disease and the number of complications were recorded. The levels of CRP,Hcy,fasting plasma glucose( FPG ),glycosylated hemoglobin(HbA1c)and blood lipid were measured. The depression group was divided into mild,medium and heavy group to compared the changes of Hcy and CRP. Results The levels of Hcy,HbA1c and the number of complications in depression group were 11. 5( 8. 6,15. 6 )μmol/L,( 10. 13 ± 2. 17 )%,and 2( 1,3 ) respectively,higher than that of non-depression group(8. 6(7. 4,11. 2)μmol/L,(9. 33 ± 2. 20)%,1(0,2)), while the education degree of depression group((9. 75 ± 3. 36)years)was lower than that of non-depression group((11. 56 ± 3. 73)years),and the differences were significant( t/Z = -3. 537,0. 952,-2. 339,0. 228 respectively;P ﹤0. 05). The levels of Hcy in mild,medium and heavy depression group were(8. 75(7. 45, 10. 45)μmol/L,12. 2(8. 90,14. 40)μmol/L,19. 50(14. 33,28. 03)μmol/L respectively and the difference was significant(F =25. 963,P =0. 000). No significance difference was found in terms of CRP level(2. 35 (1. 10,4. 92)mg/L,3. 25(1. 11,5. 68)mg/L,2. 32(1. 27,5. 41)mg/L;F=0. 194,P=0. 907). There was significant correlation between depression scores and Hcy( r=0. 615,P=0. 000). Conclusion Type 2 diabetes with depression is associated with the level of blood glucose,education degree and the course of disease. Hcy,not CRP is an independent risk factor of type 2 diabetes with depression.

OBJECTIVETo detect the relationship between point mutations on exon 2 of parkin gene and sporadic early-onset Parkinson's disease.

METHODSThe point mutations on exon 2 of parkin gene were detected using polymerase chain reaction(PCR), agarose electrophoresis, single strand conformation polymorphism(SSCP), DNA sequencing and analysis of restrict enzyme in DNA of 60 Parkinson's disease patients with an onset age under 50 and 120 normal controls.

RESULTSOne homozygous mutation (G(237)-->C) on exon 2 was found by sequencing and verified by analysis of restrict enzyme, whereas no mutation was found in normal controls.

CONCLUSIONPoint mutations on exon 2 of parkin gene are likely to be related to sporadic early-onset Parkinson's disease.

Adult ; Aged ; Aged, 80 and over ; Exons ; Female ; Humans ; Ligases ; genetics ; Male ; Middle Aged ; Parkinson Disease ; genetics ; Point Mutation ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA ; Ubiquitin-Protein Ligases