Objective To construct a lentiviral vector carrying CRKL gene RNA interfering( RNAi ).Methods The CRKL RNAi was selected and subcloned into the lentiviral vector,pGCL-GFP(including U6 promotor and green fluorescent protein),generating the lentiviral vector LV-shCRKL.The corrected CRKL was confirmed by endoenzyme digestion ,sequencing.Recombinant lentiviruses were produced by 293T cells following the eo-transfection of LV-shCRKL,with the packaging plasmids pHelper1.0 and pHelper2.0.The virus titer was detected by GFP expressions in 293T cells.Results Plasmid LV-shCRKL carried the correct sequence.The recombinant lentiviruse LV-shCRKL could be produced by co-transfection of LV-shCRKL to 293T cells.Conclusion The recombinant lentiviruse vector LV-shCRKL is constructed successful.

Objective To develop a new kind of tracheal catheter for large or medium -sized surgical anesthesia as a medical equipment of respiratory management,and to rescue the patients with anhelation of respiratory arrest,which is essential to first-aid treatment and will be widely used clinically.Methods The disposable non-toxic transparent plastic was adopted,which was reinforced by the spiral stainless steel wire with regular separations.The diameter of the tracheal catheter varied with the different models.The tracheal catheter was composed of the items put into the trachea when applied for anesthesia such as the inflatable bladder and liquid mouth,the ones connected with the anesthesia machine such as the ring lines regulating direction of the front end of the catheter,and the syringe adapters for the liquid and gas.Results The flexible,air-proof tracheal catheter,with its direction adjustable,injected the medicine through its front end.Conclusion The tracheal catheter can be used for intratracheal medication,and will be popularized clinically.

ObjectiveTo study the curative effects of using oxygen treatment combined mixture to Ⅲ-Ⅳ bedsore. Methods Divided 63 patients into the observation group (32 cases) and the traditional group (31 cases) randomly. Oxygen treatment combined mixture was used in the observation group and the traditional nursing measures were used in the traditional group. Compared the curative effects of bedsore between the 2 groups.Results The cure rate of Ⅲ-Ⅳ bedsore in the observation group was significant higher than that of in the traditional group,P

Objective:To observe the change of BDNF level expression in experimental in hippocampal and habenular nucles in experimental depression rats.Methods:The depression model of rats was established by chronic unpredictable mild stress. Openfield test was performed to detect the behavior of rats. Immunohistochemistry was used to observe the changes of BDNF level.Results:Compared to the control group,the number of depressed animals was much less than that of normal animals.Conclusion:The level of BDNF decrease significantly in experimental depressive model of rats.

Objective:To study the influence of “Warming Yang and Nourishing Qi”Acupuncture in the expression levels of acetylcholine receptor protein (AchR)in gastrocnemius neuromuscular junction (NMJ)of the rats with experimental autoimmune myasthenia gravis (EAMG),and to clarify the mechanism of “Warming Yang and Nourishing Qi”Acupucture in the treatment myasthenia gravis (MG).Methods:The female Lewis rats were used to establish the EAMG rat models immuned with AchRα1 129-145 polypeptide fragments.30 EAMG rats were selected randomly and divided into model control group, drug control group, and acupuncture treatment group (n=10),and another 10 rats purchased at the same period were used as blank control group.The rats in drug control group were treated with 1 8.5 mg · kg-1 · d-1 pyridostigmine by gavage. And the rats in acupuncture treatment group were treated with irritating by“Warming Yang and Nourishing Qi”Acupuncture.The rats in blank control group and model control group were not treated .The Lennon symptom scores of the rats in various groups were recorded.After treatment,the tissues of gastrocnemius NMJ of the rats were taken,and the AchR protein expression levels in NMJ of the rats in various groups were detected by Western blotting method. Results:Compared with blank control group,after successfully modelling,the Lennon symptom scores of the rats in model control group, drug control group and acupuncture treatment group before treatment were significantly increased (P<0.01).Compared with model control group,the Lennon symptom scores of the rats in drug control group and acupuncture treatment group after two treatment course were significantly increased (P<0.01 ). Compared with blank control group, the AchR protein expression level in gastronemus NMJ of the rats in model control group was significantly decreased (P<0.01). Compared with model control group, the AchR protein expression levels in gastronemus NMJ of the rats in acupuncture treatment group and drug control group after treatment were significantly increased (P<0.01).Conclusion:“Warming Yang and Nourishing Qi”Acupuncture has good therapeutic effect on the EAMG through increasing the NMJ AchR protein expression level of the EAMG rats.

ObjectiveTo evaluate the impacts of pereutaneous nephrolithotomy (PCNL) on perioperative renal hemodynamics.MethodsThe hemodynamics of operated renal arteries of 30 patients who underwent unilateral PCNL with single pole access were observed 1 day before and 5-7 days after operation with CDFI.Parameters were analyzed statistically.ResultsAfter operation,resistance index (RI) of renal arteries decreased (P＜0.05).The diastolic flow statistically increased at main renal artery (MRA) of renal hilus,interlobar renal artery and interlobular renal artery (all P＜0.05).After PCNL,in serious hydronephrosis patients,RI decreased (P＜0.05) at segmental renal artery (SRA) and interlobar artery,end-diastolic flow velocity (Vmin) increased at interlobar renal artery (P＜0.05).In moderate hydronephrosis patients,RI decreased at all renal arteries (P＜0.05) after PCNL,Vmin increased at MRA and interlobular renal artery (P＜0.05).In minor hydronephrosis patients,RI decreased at MRA and SRA,Vmin increased at SRA.In patients without hydronephrosis,RI changeed like serious hydronephrosis patients.ConclusionAfter PCNL,ipsilateral renal perfusion improves,renal diastolic flow increases and RI decreases.CDFI can be used to observe the blood perfusion of kidney,and provide quantitative information of renal hemodynamics.

Objective To investigate the phenotypes and genotypes of extended -spectrum β-lacta-mases (ESBLs) and AmpC β-lactamases produced by cefoxitin-resistant Shigella strains isolated from Zhe-jiang province and the virulence of those strains .Methods Kirby-Bauer antibiotic testing was performed to screen cefoxitin-resistant strains from 356 Shigella isolates.The serotypes of the cefoxitin-resistant strains were detected .The phenotypes of ESBLs and AmpC β-lactamases from cefoxitin-resistant strains were detec-ted by ESBLs confirmatory test and AmpC-three-dimensional test ,respectively .The genotypes of ESBLs and AmpC β-lactamases were analyzed by PCR and sequence analysis .The virulence genes ( virA, ial, iapH, set1A, set1B and sen) in the cefoxitin-resistant strains were screened by PCR .Results Eight cefoxitin-re-sistant strains were identified from 356 Shigella isolates.Among them,six strains were identified as Shigella flexneri (S.fleaneri) strain (four F2a,one F2b and one F2c),and the rest were identified as Shigella sonnie (S.sonnei) strain.Among all eight cefoxitin-resistant strains,five strains showed positive results for ESBLs confirmatory test and two strains showed positive results for AmpC-three-dimensional test .Seven out of the eight strains harbor ESBLs genes (CTX-M-14,15,57 and/or OXA-30),two of which were positive for AmpC genes (DHA-1 and CMY-2).Five out of the eight cefoxitin-resistant strains carried all of the six tested viru-lence genes,while the other three strains possessed four virulence genes except for set1A and set1B.The two strains producing both ESBLs and AmpC β-lactamases were susceptible only to imipenem .Conclusion ESBLs positive isolates are prevalent strains of cefoxitin-resistant Shigella with potential high virulence .Some of them also produce AmpC β-lactamases ,and the DHA-1 type of AmpCβ-lactamase is identified for the first time in China .

Objective To detect the antibodies of streptococcal C5a peptidase from group B streptococcus(SCPB)in neonates,demonstrate the existence of SCPB antibody in pregnant women after natural group B streptococcus(GBS)infection,and provide clinical evidence for prevention of GBS infection. Methods Sera were collected from 107 neonates(80 term infants and 27 premature infant)between February 2007 and December 2007. The antibodies of SCPB were detected using ELISA method,and cultures of GBS were done simultaneously. Results 21(19.6%)newborns were found to be SCPB antibody positive(including 20 term infants and 1 premature infant),the difference of positive ratio between term and premature infant was significant(25% and 3.7%,respectively). Conclusions This study indicated that pregnant women could produce SCPB antibody by immune response,and transmitted it to the infants through the placenta. Further study is needed to clarify the effect of SCPB antibody in expectant mother and newborn with GBS infection.

Objective:To evaluate the therapeutic methods of sensorineural hearing loss with iron deficiency anemia and their effects. Method:Sixty-eight Wistar rats were randomly divided into 3 groups: Group A (iron deficiency group A) 24 rats, group B (iron deficiency group B) 24 rats and group C (control group) 20 rats. Group A and B were fed with iron deficiency diet for 6 weeks. By analyzing ABR and DPOAE, hearing loss was detected in group A and B. Then,group A was given iron supplementation diet and hyperbaric oxygen therapies. and group B was given iron supplementation diet only. Result:The auditory thresholds and period latencies (PL) of I waves of ABR and DPOAE amplitude in 2 kHz and 3kHz within group A and B were significantly different before and after treatment (P＜0.05). But the effects of treatment between these two groups had not significant differences (P＞0.05). Conclusion: It suggested that good results can be obtained by early iron supplementation in iron deficiency hearing loss cases. Hyperbaric Oxygen therapy may not be necessary.

ObjectiveTo investigate the consistency between the results of passive particle agglutination test ( PPA ) and ELISA on Mycoplasma pneumonia ( M.pneumonia ) infection.Study the diagnostic value of both assays.Methods From November 2010 to May 2011,the serum samples of ]191 patients with respiratorysymptoms were collected fromAffiliatedHospitalof JiningMedical University.All samples were tested for antibody levels against M.pneumonia using PPA,and for IgM,IgG,IgA subclass using ELISA.The correlation between the results of two methods was evaluated by Kappa test and Spearman rank correlation analysis.The variances of the antibody subclasses among samples with different PPA titers and different age groups were analyzed by Kruskal-Wallis test.The infection status of patients was analyzed based on ELISA results and the clinical relevance of both assays was evaluated in comparison with clinical diagnosis for samples with high PPA titer.ResultsThe level of agreement between the results of PPA and ELISA was 84.3％,with Kappa value of 0.642 ( P ＜0.01 ).The prevalence of IgM and IgA antibody against M.pneumonia was significantly different among samples with various PPA titers ( P ＜0.05 ).The prevalence of IgM subclass was higher in chill and teeuager groups,while that of IgA and IgG were higher in elderly group.Antibody isotyping results suggested that 58.1％ of PPA positive samples (75 cases),especially 96.4％ of samples with high PPA titer ( 27 cases),were of current infection,which was in consistent with clinical diagnosis.ConclusionPPA showed good consistency with ELISA on diagnosis of Mycoplasma pneumonia infection.Antibody subclass determination hy ELISA indicates disease progression,thus to differentiate current infection from past.