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 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 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 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 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 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 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 explore the predictive value of serum thyroglobulin autoantibody (TgAb) for recurrence / metastasisin of thyroglobulin (TG)-negative and TgAb-positive patients with differentiated thyroid carcinoma(DTC) after thyroid ablation.Methods:The clinical data of 57 patients with DTC,undergoing complete resection of thyroid tissue,TG negative and TgAb positive,who were reviewed in Liaocheng People's Hospital during April 2013 to April 2015,were selected and divided into recurrence/metastasis group (n=20) and no recurrence/metastasis group (n=37).The TG and TgAb levels in the two groups were detected and compared by the electrochemical luminescence method,the sensitivity,specificity,positive predictive value and negative predictive value of TgAb in the diagnosis of DTC recurrence / metastasis were analyzed;the independent risk factors of DTC recurrence / metastasis were analyzed by Logistic regression.Results:The TgAb levels(72～3850 IU/mL) in the recurrence/metastasis group was higher than that(18～3638 IU/mL) in the no recurrence/metastasis group,the difference was statistically significant (P＜0.05).The sensitivity,specificity,positive predictive value,negative preictive value of TgAb in the diagnosis of recurrence/metastasis of DTC were 85.71％,83.33％,75.00％,90.91％,respectively.Logistic Regression analysis showed that the 100≤ TgAb ＜204 IU/mL,204≤ TgAb≤ 1000IU/mL,and ＞ 1000IU/rnL levels of TgAb were the independent risk factors of rucurrence/metastasis ofDTC (OR=1.267,2.853,6.791,P＜0.05).Conclusion:TgAb can be used as evaluation of an important index of the recurrence/metastasis of patients with DTC when serum thyroglobulin (TG) was negative and TgAb was positive after thyroid ablation.The higher the TgAb levels,the more probability of the recurrence/metastasis.

Objective To evaluate the effect of obesity on the potency of propofol for sedation.Methods Sixty patients of both sexes, aged 35-55 yr, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ , scheduled for elective operation under general anesthesia, were enrolled in the study.The patients were divided into 2 groups (n=30 each) according to the body mass index (BMI) : normal body weight (BMI ＜ 25 kg/m2) group (group C) and obesity (BMI 30-40 kg/m2) group (group O).No patients received premedication.Propofol was given by target-controlled infusion.The initial target plasma concentration of propofol was set at 1.2 μg/ml.After the target concentration was steadily maintained for 30 s, it was increased in 0.3 μg/ml increment until the patients lost consciousness (OAA/S score =1).The target plasma concentration of propofol was recorded during each period.The median effective concentration (EC50) and 95％ confidence interval of propofol for loss of consciousness was calculated using probit analysis.Results The EC50 and 95％ confidence interval of propofol for loss of consciousness were 3.82 (3.73-3.90) and 3.29 (3.20-3.37) μg/ml in group C and group O, respectively.Compared with group C, the EC50 was significantly decreased in group O (P＜0.05).Conclusion Obesity can enhance the potency of propofol for sedation.

Objective To investigate the emotional state and its influencing factors in children with nocturnal enuresis,so as to improve clinical attention and give appropriate intervention.Methods Totally 166 nocturnal enuresis patients aged 9 -15 years old were admitted to Beijing Children′s Hospital Affiliated to Capital Medical University from August 2011 to August 2014.Specifically,Screen for Child Anxiety Related Emotional Disorders (SCARED)and Depres-sive Self -Rating Scale for Childhood (DSRSC)were utilized to test the patients′emotional state.The comparison be-tween the general norm data of the healthy children and the psychological test of enuresis children were performed,the lat-ter emotional state and its influence factors were evaluated.Analysis of variance and t -test were performed.Frequency comparison was done by using χ2 test.Influence factors analysis was made by Logistic or multiple regression analysis. Results The incidence of children′s anxiety with enuresis was 39.2%(65 /166 cases).Anxiety overall scores of children with enuresis amounted to 21 .16 ±11 .88 which were significantly higher than the norm(14.40 ±9.41 )(P <0.05).The diagnosis rate of depression in the NE patients was 9.0%.No significant difference was found in the depression total scores(P >0.05).The emotional disorders mainly appeared in patients with primary enuresis,without family history,poor grade and poor relationships with family members and peers.The school performance was an important factor for enuresis patients with anxiety disorders(P <0.05).Conclusions Children with nocturnal enuresis are more likely to have anxiety disorders than normal children.Their emotional states is related to the enuresis type,family history,school performance, family and peer relationship.Therefore,attention should be paid to the emotional state of children with nocturnal enuresis. According to the adverse factors for nocturnal enuresis patients,effective treatment and appropriate psychological interven-tion should be given at the same time.