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 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 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.

AIM: To investigate the changes of potassium channels in thoracic aorta of streptozotocin-induced diabetic mouse in the early stage of diabetes mellitus. METHODS: The effects of 60 mmol/L KCl, phenylephrine (PE), sodium nitroprusside (SNP) were measured and concentration-response curves to SNP were determined in the presence and in the absence of the inhibitors of potassium channels on the thoracic aortic rings of diabetic and age-matched control mice in vitro. RESULTS: STZ-diabetic mice showed a significant increase in the maximum contractile response and sensitivity of thoracic aorta to 60 mmol/L KCl and PE. The endothelium-independent relaxation response to SNP was increased by diabetes and were decreased significantly by pretreatment of the vessels with 1 mmol/L tetraethylammonium (TEA), 1 mmol/L 4-aminopyridine (4-AP) and 10 ?mol/L glibenclamide in diabetes thoracic aorta. Only 4-AP decreased relaxation response to SNP in age-matched control mice. The -logIC 50 difference of TEA in thoracic aorta rings of diabetes was significantly higher than age-matched control mice.CONCLUSION: In early stage of diabetes mellitus, the opening or expression of K Ca channels is significantly enhanced.The opening of K ATP channels is also enhanced in this stage.

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.

Background and purpose: Peritoneal metastasis of gastric cancer is mainly discovered in the ad-vanced cancer. Nonetheless, the clinical applicability of each tumor biomarker in peritoneal metastasis of gastric cancer is still ambiguous. Therefore, this study investigated the diagnostic value and clinical significance of CEA, CA125 and CA72-4 in gastric carcinoma patients with peritoneal metastases. Methods: A total of 108 gastric carcinoma patients with peritoneal metastases from Jan. 2008 to Dec. 2013 were studied. All patients were diagnosed by imaging, operations and pathological examination, and also received intravenous or intraperitoneal chemotherapy. Serum tumor markers such as CEA, CA125 and CA72-4 were determined during diagnosis and before each chemotherapy. The diagnostic sensitivity of single marker and combined detection with 2 or 3 markers were analyzed. The correlations among the serum tumor markers and clinical pathological factors, chemotherapeutic effects and survival time were analyzed. Results: Positive rates of CEA, CA125 and CA72-4 were 20.4%, 46.3% and 45.4% in gastric cancer patients with peritoneal metastases, respectively. For these patients, the positive rates of CEA/CA125, CEA/CA72-4, CA125/CA72-4 and CEA/CA125/CA72-4 were 54.7%, 52.8%, 69.5% and 79.6%, respectively. The combined detection of 3 tumor markers was much better than single marker detection (P<0.05). Positive rates of CEA, CA125 and CA72-4 were correlated with the ECOG scale (P<0.05). Positive rate of CA125 was associated with ascites (P<0.001), while positive rate of CA72-4 was associated with ovarian metastasis (P<0.05). Median survival time of patients with positive rates of CEA, CA125 and CA72-4 was significantly lower than that of the patients with normal levels of these markers (P<0.05). Compared with pre-treatment, the levels of all three tumor markers significantly declined after three cycles of chemo-therapy (P<0.05). The decline in CA125 level after chemotherapy was significantly correlated with decreased amount of ascites (P<0.05). The tumor markers turned negative after 3 cycles chemotherapy in patients with positive markers upon initial diagnosis, their survival was significantly prolonged (P<0.001). Conclusion: Combined detection of serum CEA, CA125 and CA72-4 can significantly promote diagnostic rate of gastric cancer with peritoneal metastasis, and may be helpful in evaluating chemotherapeutic effects and predicting prognosis.

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.