Bacillus megaterium strains are commonly used in microbial fertilizer(MF) . MF products are often contaminated by other B. cereus group members,which have similar phenotype such as Bacillus cereus,B. thuringiensis,B. mycoide. For quality control and safety of MF,a rapid and accurate method is needed to distinguish the strains of Bacillus megaterium from B. cereus group. Based on specific nucleotide sequences of the spoOA genes,2 pairs of species-specific primers were designed and a multiplex-PCR(mPCR) was developed for this purpose. When the optimized mPCR was used to detect the DNAs of 24 reference strains from three genera of Bacillus,Paenibacillus,and Brevibacillus,all B. megaterium strains showed singlefragment of 443 bp and Bacillus cereus group showed a fragment of 411 bp. However,no any amplified product was from the other bacteria. The sensitivity of mPCR was 105 CFU/mL. The mPCR results of 10 isolates of B. megaterium/B. cereus group and 8 products of MF coincided with the biochemical assay. Taken together,our newly developed mPCR assay was species-specific and effective in application. It can be used to detect and identify the strains of B. megaterium and B. cereus group from microbial fertilizer products.

OBJECTIVE To investigate the conditions and associated risk factors of nosocomial infections caused by Stenotrophomonas maltophilia.METHODS Thirty isolates of S.maltophilia causing nosocomial infections were collected and identified with API 20NE test strips.Minimal inhibitory concentration(MIC) of 14 antimicrobial agents against 30 isolates was determined by broth microdilution method.Case-control study and multivariate Logistic regression analysis were used for statistics to verify risk factors of infections caused by S.maltophilia.RESULTS Thirty isolates of S.maltophilia were highly resistant to imipenem,meropenem,cefotaxime, aztreonam and amikacin,but showed certain susceptibility to cefoperazone/sulbactam,piperacillin/tazobactam,trimethoprim-sulfamethoxazole and ticarcillin/clavulanic acid(96.7%,76.7%,73.3% and 60.0%,respectively).The independent risk factors leading to infections of S.maltophilia were mechanical ventilation(OR=7.629) and over 60 days of length of stay(OR=4.466).CONCLUSIONS S.maltophilia shows multiresistance to commonly used antimicrobial agents.The mechanical ventilation and over 60 days of length of stay are the independant risk factors for nosocomial infections caused by S.maltophilia.

Objective To investigate the efficacy and safety of dipeptidyl peptidase 4(DDP-4)inhibitor in the treatment of diabetes mellitus.Methods Eighty-six cases patients with poor glycemic control in type 2 diabetes mellitus in Shunde First People's Hospital of Foshan from May 2015 to May 2016 were selected as the research objects and divided into two groups according to random number table method,each group with 43 cases.The control group with acarbose(50 mg/times,3 times/d,with the same as the 3 meals,sustained medication for 12 weeks)to control blood sugar,while the observation group used DPP-4 inhibitors sitagliptin(100 mg/times,1 times/d,sustained medication for 12 weeks),compared the blood glucose,blood lipid indexes and adverse reactions of two groups before and after treatment.Results After treatment,FPG,2 hPG,HbAlc level of observation group were respectively(6.71±0.65)mmol/L,(8.10±0.17)mmol/L,(7.12±0.41)％,significantly lower than control group((7.86±0.72)mmol/L,(9.20±0.65)mmol/L,(7.51±0.52)％,the differences were statistically significant(P＜0.01).After treatment,there were no significant differences in terms of TC,TG,HDL-C,LDL-C between two groups(P＞0.05).Treatment for 12 weeks,the blood glucose compliance rate of observation group was 88.37％(38/43),the control group was 67.44％(29/43),the observation group was significantly higher than the control group(P=0.019).After treatment,the HOMA-IR value of the observation group was 4.42±0.17,significantly lower than the control group(4.91±0.24),HOMA-beta value was 88.20±6.31,significantly higher than that of the control group(80.21±5.67),the differences were significant(P＜0.01).Conclusion DDP-4 inhibitor in the treatment of type 2 diabetes,can effectively reduce the level of blood glucose and glycosylated hemoglobin,and have no significant adverse reactions,is effective and safe hypoglycemic drugs.

Objective To evaluate the effects of dexmedetomidine on the expression of tumor necrosis factor-α(TNF-α) and intercellular adhesion molecule-1 (ICAM-1 ) during cerebral ischemia-reperfusion (I/R ) in rats .Methods Thirty-six male Sprague-Dawley rats , aged 8-12 weeks ,weighing 280-320 g ,were randomly divided into 3 groups ( n= 12 each ):sham operation group (group S ) , cerebral I/R group (group I/R ) and dexmedetomidine group (group D ) .The animals were anesthetized with 10% chloral hydrate 3.5 ml/kg .The focal cerebral ischemia was induced by middle cerebral artery occlusion using a nylon thread with rounded tip inserted into internal carotid artery and advanced cranially until resistance was met .In group D ,dexmedetomidine 3μg/kg was injected intravenously immediately after beginning of ischemia ,followed by infusion at 3μg·kg-1 ·h-1 for 120 min ,while the equal volume of normal saline was given in group I/R .Four rats from each group were chosen at 24 ,48 and 72 h of reperfusion , and blood samples were taken from the left ventricle to determine the concentrations of serum S100B protein .The rats were then sacrificed and the brains were removed to determine the expression of TNF-αand ICAM-1 in brain tissues .Results Compared with group S ,the concentrations of serum S100B protein were significantly increased , and the expression of TNF-α and ICAM-1 in brain tissues was up-regulated in I/R and D groups ( P<0.05) .Compared with group I/R ,the concentrations of serum S100B protein were significantly decreased , and the expression of TNF-α and ICAM-1 in brain tissues was down-regulated in group D ( P<0.05 ) .Conclusion The mechanism by which dexmedetomidine reduces the cerebral I/R injury is related to down-regulation of TNF-αand ICAM-1 expression in rats .

Optically pure α-substituted propanoic acids and their derivatives represent as an important kind of organic building blocks and key intermediates,which has been widely used in the synthesis of chiral drugs.Some of them have been used directly as nonsteroidal anti-inflammatory drugs (NSAIDs),such as ibuprofen,naproxen,ketoprofen and so on.Dihydroartemisinic acid (DHAA),the same structure as the α-substituted propanic acids,is a key intermediate for the synthesis of artemisinin,the most effective and current used anti-malarial drug.Therefore,the asymmetric synthesis of α-substituted propanoic acids is always a hot topic for chemical scientists.Asymmetric catalytic hydrogenation attracts more and more attentions because of its atom economy and efficiency.This dissertation will disclose the asymmetric synthesis of α-substituted propanoic acids using transition metal-complex as a chiral catalyst.

Objective To observe the effect of the soft tissues injury around atlantoaxial joint on memory abilities of mice.Methods 44 mice were divided into the model group and control group with 22 animals in each group.The soft tissues around atlantoaxial joint were injured in the model group by surgery between the first and the second cervical vertebrae.The changes of learning and memory of mice were observed with step-down avoidance test,water-maze test and hole-board test;the expression of caspase-3 in hippocampus was observed by immunohistochemical staining.Results Compared with the control group,escape latency was longer and wrong times were more in water-maze test of the model group(P<0.01). In step-down avoidance test of the model group,the reaction period prolonged and wrong times increased obviously(P<0.01).In hole-board test,the latency period prolonged and the times of extending the holes decreased significantly(P<0.01).Degenerated or dead neuron in hippocampus of surgery-treated mice was found scatteredly or continuously with HE staining,the expression of caspase-3 protein was showed positive with immunohistochemical staining.Conclusion The soft tissues injury arround atlantoaxial joint can induce abnormality of the stability of cervical vertebra,which aggravated the aging process of the mice and decreased abilities of learning and memory.

Objective To investigate the effect of preconditioning with different concentrations of sevoflurane on hypoxia-reoxygenation(H/R)-induced apoptosis in rat hippocampal neurons and the role of mitochondrial KATP(mito-KATP)channels.Methods Primary cultured hippocampal neurons isolated from newborn SD rats(＜24h)of both sexes,weighing 5-6 g,were randomly divided into 7 groups with 48 wells and 12 dishes in each one:control group(C group),H/R group,preconditioning with 6%,4%and 2% sevoflurane groups(S1-3 groups),5-hydroxydecanoate(5-HD,mito-KATP channel blocker)100 μmol/L preconditioning group(5-HD group)and preconditioning with 5-HD 100 μmol/L+6% sevoflurane group(5-HD+S group).The neurons were exposed to 4 h hypoxia followed by 24 h reoxygenation. In S1-3 groups, preconditioning was performed with 6% , 4% and 2% sevoflurane respectively before H/R. In 5-HD group, preconditioning was performed with 5-HD (final concentration 100 μmol/L) before H/R. In 5-HD + S group, preconditioning was performed with 5-HD 100 μmol/L and 6% sevoflurane before H/R. The neuronal viability, apoptosis rate and expression of Bcl-2 and Bax were determined after 24 h reoxygenation.Results The neuronal viability was significantly lower,while the apoptosis rate and expression of Bcl-2 and Bax were significantly higher in the other 6 groups than in group C(P＜0.01).The neuronal viability and expression of Bcl-2 were significantly higher,while the apoptosis rate and Bax expression were lower in S1-3 groups than in group H/R. There was no significant difference in the parameters mentioned above between 5-HD and 5-HD + S groups(P＞0.05).The neuronal viability and expression of Bcl-2 were significantly lower, while the apoptosis rate and Bax expression were higher in S2, S3 and 5-HD + S groups than in group S1, and in group S3 than in group S2(P＜0.0l) .Conclusion Sevoflurane preconditioning can inhibit H/R-induced apoptosis in rat hippocampal neurons and reduce the injury to neurons in a concentration-dependent manner, and the underlying mechanism may be related to activation of mito-KATP channels, up-regulation of Bcl-2 expression and down-regulation of Bax expression.

Objective Transesophageal Doppler monitoring ( TDM) has been widely applied to fluid therapy in some major op-erations but rarely used in aortic valve replacement.The purpose of this study was to assess the value of TDM in fluid therapy, vasoactive drugs use, and left ventricular function in patients undergoing aorta valve replacement after cardiopulmonary bypass. Methods This study included 40 patients undergoing aotic valve replacement after cardiopulmonary bypass in Anhui Provincial Hospital from March 2014 to June 2015, which were randomly assigned to a TDM and a control group of equal number.TDM was used for the guidance of fluid infu-sion and vasoactive drug administration for the patients in the former group, while central venous pressure, mean arterial pressure, and heart rate were employed for those in the latter.The fluid volume infused, urine volume, blood loss, hemodynamics, arterial blood lac-tate, and outcomes were compared between the two groups. Results Compared with the control, the TDM group showed significantly increases in the colloidal fluid volume ([303 ±60] vs [373 ±65]mL, P<0.05), crystal fluid volume ([533 ±87] vs [596 ±83]mL, P<0.05), and urine volume ([274 ±95] vs [338 ±84]mL, P<0.05), but remarkable decreases in the red blood cell count, blood plasma vol-ume, postoperative extubation time, doses of dopamine and dobu-tamine, and length of hospital stay (all P<0.05).The hemodynamics were more stable and arterial blood lactate was lower in the TDM group than in the control (P<0.05). Conclusio n Transesophageal Doppler monitoring can timely and exactly manifest the hemodynamic changes and left ventricular function of the patient undergoing aotic valve replacement and provide precise guidance for fluid therapy and vasoactive drug administration.

Objective To investigate the value of plasma miR-22 in diagnosis of idiopathic pulmonary arterial hypertension ( IPAH ),and its role of regulation mechanisms in the pathogenesis of the disease.Methods Circulating miR-22 levels of IPAH patients and healthy controls were evaluated by RTPCR.The silico analysis of targets for miR-22 was taken, and followed by eGFP reporter assay for verification of predicted target gene Myc binding protein (MYCBP). Results Compared with healthy controls,the expression of plasma miR-22 in IPAH patients was significantly decreased (P ＜ 0.01 ).The area under curve (AUC) of ROC curve was 0.744.MYCBP was a real target of miR-22 confirmed by silico analysis and eGFP reporter assay. Conclusions The expression of plasma miR-22 was significantly decreased in IPAH patients,and it could serve as a potential biomarker for diagnosis.The miR-22 might be involved in the pathogenesis of the disease through promoting its target gene MYCBP to activate the c-Myc pathway.

Objective To explore the relevance between nitrate transporters (Sialin) and ischemia-reperfusion injury (IRI) in free flaps.Methods Twenty rats were randomly divided into control group and experimental group,10 in each group.After preparatiing 4 cm×8 cm free tissue flap on left lower abdominal,the rats in experimental group were experienced ischemia in 6 hours,then reperfusion.The rats in control group were without ischemia treatment,and tissue flaps were sutured in situ.After postoperative observation for 7 days,the furthest edge of survival flaps were sampled.The expression of Sialin was detected by the Real-time PCR and Western blot.The change of nitrate content was analysed by chemiluminescence.Results Both of results of Real-time quantitative PCR and Western blot,it was found that expression levels of Sialin in the experimental group were significantly lower than the control group(0.945±0.0530)(P＜0.05).Applying Real-time quantitative PCR relative quantitative analysis,it was found that the experimental group Sialin mRNA expression level was 0.284±0.0486,significantly lower than that of control group (0.945±0.0530)(P＜0.05).Using Western blot technical analysis,experimental Sialin protein expression level was(0.1413±0.0446),significantly lower than the control group (0.3519±0.0368) (P＜0.05).The concentration of nitrate of test results found that the expression of nitrate transporters (Sialin) linearly related with the concentration of nitrate in free flaps,which means tbe Sialin expression level decreased,the nitrate content decreased(P＜0.05,R=0.81).Conclusion The concentration of nitrate was linearly related to the level of Sialin expression:when Sialin expression levels dropped,the concentration of nitrate decreased.Nitrate transporters(Sialin) may be correlated with IRI in free flaps.