Suppression subtractive hybridization(SSH),which is feasible and highly specific,is one of the most efficient methods for cloning differentially expressed genes.It has been successfully applied in the study of tumors,stem cells,etc.

Objective To develop a HPLC-MS/MS methodfor determination of cyclosporin A(CSA)and AM1 in bone marrow transplant patient,and explore the relationship of CSA and its main metabolite AM1 within individual and between individuals,and provide reliable basis for clinical rational use of monitoring in CSA blood drug concentration.Methods CSD was used as internal standard,and whole blood samples were treated with internal standard methanol precipitated protein.The column was Ultimate XB-C18 with a column temperature of 65 ℃ and the mobile phase was eluted with 0.1％ of formic acid and 2 mmol/L ammonium acetate in water and methanol containing 0.1％ formic acid.The mass spectrometry was detected by electrospray ion trap positive ion mode,MRM scanning,monitoring CSA 1219.9 to 1203.1 m/z,AM1 1236.1 to 1219.1 m/z,CSD 1234.0 to 1217.0 m/z.Results The concentration of CSA was linear in the range of 16 to 1600 ng/mL,Y=0.0143X+0.0213(r=0.9976).The concentration of AM1 was linear in the range of 10～1000 ng/mL,Y=0.00363X-0.00528(r=0.9973).The ratio of CSA to AM1 (AM1/CSA) between individual ranged from 32％ to 356％.The ratio of CSA to AM1 within indiviolual(AM1/CSA) ranged from 27 ％ to 147 ％.Conclusion HPLC-MS/MS method for the simultaneous detection of CSA and AM1 was established.The variation of CSA exists in the bone marrow transplant patient between individuals and within individual;the HPLC-MS/MS method can be used for monitoring of whole blood concentration of CSA in bone marrow transplant patients.

Objective To estabolish the ALDH2 Glu504Lys genotyping technique for oral swab sample type based on PCR-gold magnetic nanoparticle chromatographyl.Methods According to the ALDH2 Glu504Lys gene the specific primers were designed by ARMS-PCR technique.Optimized the optimal ALDH2 Glu504Lys oral swab reaction system and PCR reaction procedures based on the routine reaction conditions and reaction procedure of laboratory PCR.60 samples of oral swabs were collected and tested by gold magnetic nano-chromatography.The results were verified by sequencing.Results The detected 60 cases of oral swab samples contain 16 cases heterozygous mutation,wild type 42 cases,homozygous mutation 2 cases and then the results by Goldmag nanoparticle detection was perfectly matched with sequencing results.Conclusion The Goldmag nanoparticles chromatographic detection technique of ALDH2 Glu504Lys buccal swabs system was established.With accuracy,fastness,reliability,it is worthy of clinical promotion.

Objective To explore the clinical effect of ozone combined with nifuratel nystatin vaginal soft capsules on the treatment of vaginitis, and to provide the basis for clinical application. Methods 204 patients with vaginitis in our hospital from January 2013 to December 2016 were divided into the observation group (114 cases) and the control group (90 cases) according to the different treatment methods. The control group was treated with nifuratel nystatin vaginal soft capsules, and the observation group was treated with ozone combined with nifuratel nystatin vaginal soft capsules. The clinical efficacy, symptom relief time and adverse reaction were compared between the two groups. Results After treatment, the total effective rate in the observation was 97.37%, which was significantly higher than 85.56% in the control group(P<0.05). The rapid remission rate of the clinical symptoms in the observation group was significantly higher than that in the control group (P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups.Conclusion The effect is significant that ozone combined with nifuratel nystatin vaginal soft capsules is used on the treatment of vaginitis, which can quickly relieve symptoms, and will not significantly increase the adverse reactions, is worthy of application.

Objective To explore the prevalence of depression in diabetes foot ulcer patients , and to an-alyze the correlation between depression morbidity and various factors such as age , HbA1c level, and quality of life etc.Methods 73 diabetes foot ulcer patients admitted from Sep .2012 to Sep.2014 were enrolled.Depres-sion was assessed using the 9-item patient health questionnaire (PHQ-9).Data such as the age, sex, duration of diabetes and foot ulcer , HbA1C level, smoking were recorded .The severity of foot ulcer was evaluated by Wagner's classification.The quality of life was scored by short form(SF-36)health survey questionnaire.Results The morbidity of depression in diabetes foot ulcer patients was 49.3%in total and 17.8% in the moderate to severe depression group .The prevalence in patients with HbA 1c≥7.0%was significantly higher than that in pa-tients with HbA1c<7.0%(66% vs 35%,P<0.01).Higher occurance of HbA1c≥7.0% was found in the moderate to severe depression group compared to that in the mild depression group .The prevalence of depression was 39.1%in grade 1 in wagner classification , 48.6% in grade 2 to 3 in wagner classification and 69.2% in grade 4 to 5 in wagner classification , showing the two was positively correlated .When duration of foot ulcer lasted more than 6 months, the morbidity of depression(58.1%)was significantly increased(P<0.05).The quality of life was significantly declined in patients with depression by SF-36 questionnaire ( P<0.05 ) .No association was found between the depression morbidity and the age , duration of diabetes , amount of complications or smoking (P>0.05).Conclusions This study shows a high prevalence of depression symptom in diabetes foot ulcer pa -tients.Depression is associated with HbA1c level, the extent and duration of foot ulcer .

Objective To apply the theory basis for effectively preventing and treating rotavirus infection by studying on specific humoral immune response in children with rotavirus diarrhea. Methods Seventy-five patients of children (diarrhea group ) and 45 controls (control group) were selected. The antigen and specific antibody in plasma and stool were detected by ELISA method;the type of viral gene was confirmed by reverse transcription PCR method;the expression of mRNA in peripheral blood mononuclear cell was measured by fluorescent light quantitation PCR method. Results In diarrhea group, rotavirus G typing to G3 dominated, accounting for 77.3% (58/75), P sub-type to P8 mainly, accounting for 82.7% (62/75). Rotavirus-specific antibody titers of plasma in diarrhea group were significantly higher than those in control group, IgA antibody titers of stool increased significantly, not only higher than that in control group and plasma. The percentage of CD19+ cells in diarrhea group [(30.8 ± 7.9)%] was significantly higher than that in control group [(23.1 ± 7.7 )%] (P = 0.009 ). And the proportion of CD4+ cells decreased significantly (P = 0.005 ), the expression level of IL- 12p40 mRNA in the entire process in diarrhea group was significantly higher than that in control group(P＜ 0.01 ). Conclusion Children with acute rotavirus infection early in the immune response to specific humoral immunity, especially mucosal immune to significantly improve the main features.

With accelerated population aging, traditional pension mode cannot adapt the diversified medical and nursing requirements of the elders. Therefore, it should reposition the function of medical service in pension career and encourage combining medical service with pension service. Through the methods of setting medical insti-tutions in pension institutions with conditions, developing social pension service in medical institutions, pension in-stitutions cooperating with medical institutions,both professional medical service and basic pension service would be offered to the elders, and thus to realize the integration of medical and nursing resources and to improve health con-dition of the elders.

Objective To develop a HPLC-MS/MS method for determination of Tacrolimus(FK506) in bone marrow trans plant patient,and explore the relationship of HPLC-MS/MS with CMIA and Elecsys in the measurement of FK506 blood drug concentration,and provide reliable basis for clinical rational use of monitoring method in FK506 blood drug concentra tion.Methods The separation was performed on a Ultimate xB-C18 column with a mobile phase of water (containing 2 mmol/L ammonium acetate and 0.1 ml/dl formic acid) and methanol (containing 0.1 ml/dl formic acid).The way of eluting was gradient.Mass spectrum detection method was ESI positive ion mode and the MRM transitions of FK506 m/z 821.5～ 768.5 and Ascomycin m/z 809.4～756.4.Selected part of the whole blood samples of FK506 in bone marrow transplant patient and respectively were tested by HPLC/MS/MS method,CMIA method and Elecsys method.Then,three methods for the detection of FK506 density were compared.Results The standard curve of FK506 was linear over the range of 0.5～50 ng/ml,Y=0.228X-0.003 08 (r=0.999 0).FK506 blood concentrations were not significantly different in HPLC/MS/MS method,CMIA method and Elecsys method.Conclusion The HPLC-MS/MS method in the detection of FK506 was established and can be used for monitoring of whole blood concentration of FK506 in bone marrow transplant patients.

Objective To explore the expression of HIF-2? and VEGF in renal cell carcinoma (RCC) and their correlation with angiogenesis and metastasis of renal cell carcinoma. Methods Expression of HIF-2? and VEGF were assessed by immunohistochemical straining in 48 cases of renal cell carcinomas. Results In 48 cases of renal cell carcinomas, positive rate of HIF-2? was 70.8%. Strong expression of HIF-2? was observed in 12 cases, moderate in 10 cases, weak in 12 cases and no expression in 14cases.There was a significant difference in HIF-2? expression between carcinoma tissues and control tissues (P

Objective To investigate the influence of mycophenolate mofetil(MMF) on blood and urine biochemical indica‐tors as well as the expressions of IL‐1 in anti‐glomerular basement membrane(GBM ) nephritis SD rats .Methods The SD rats were randomly divided into control group ,model group and MMF group .And then the models of SD rat anti‐GBM nephritis were estab‐lished by injecting rabbit anti‐SD rat GBM serum antibodies .The urinary protein and blood parameters were detected initially ,1st , 2nd ,3rd and 4th weeks after administration respectively .The expressions of IL‐1 and TGF‐β1 were detected by Elisa Kit and the re‐nal pathological changes were observed by HE and PAS staining .Results After treatment for 4 weeks by MMF ,the 24 h UP ,BUN and SCr levels were lower in MMF group than those of the model group(P<0 .05) .And the expressions of IL‐1 and TGF‐β1 were decreased in MMF group .The results of HE and PAS staining showed that in model group rats ,glomerular tuft morphology was destroyed and inflammatory cell infiltration was observed in the glomeruli .Some glomeruli had cellular crescent formation .Infiltra‐tion of inflammatory cells was also observed in the interstitium .MMF clearly improved these pathological changes overall ,with im‐provements noted in both glomerular and tubular degeneration as well as in the infiltration of inflammatory cells .Conclusion MMF shows certain renal protective effect on anti‐GBM nephritis SD rats .