Objective:To investigate the effect of the titin gene and its influence on the contraction protein gene such as ?-MHC and cTnI in the procession of MSCs differentiating into cardiomyocytes in vitro.Methods:MSCs were isolated from bilateral thighbones and tibias of Wistar rats,purified、expanded and then exposed to 5-aza.we observed the morphology of MSCs with phase-contrast microscope at 1st,4th 7th,14th day after induction.Then we transfected the recombination plasmid vector(including pSITITIN and pSIACTIN)into MSCs after induction.detected the expression of titin、?-MHC and cTnI at interval times by RT-PCR.Results:1.we obtained the mild expression of titin、?-MHC and cTnI gene before 5-aza treament,but the expression of them increased after induction.2.After recombination plasmid were transfected into MSCs which have been induced by 5-aza,the expression of titin gene was decreased 3.The expression of ?-MHC gene kept invariant,while the expression of cTnI gene was weakened after the interference.Conclusion:1.titin gene probably play an important role in the procession of MSCs differentiating into cardiomyocytes in vitro.2.titin gene may regulate the expression of cTnI.

Objective To investigate the Clinical Value of Combined Determination of Serum cystatin C and Retinol-binding protein in diabetic nephropathy.Methods Immunoturbidimetry was employed to analyze Cystatin C(Cys C)and Retinol-binding protein;(RBP) from 94 cases of diabetic patients and 110 healthy control group .and we also detected thevalue of serum creatinine (Scr),urinary creatinine (Ucr),and calculated creatinine clearance rate (Ccr),compared the correlation of Cys C,RBP with Ccr,Scr.Results The contents of serum Cys C,RBP in the group of diabetic nephropathy were significant higher than that in healthy controllers(P<0.01),and Cys C,RBP were significantly negative correlation with Ccr.Conclusion The sensitivity and accuracy of the combined determination of serum Cys C,RBP in diabetic nephropathy are improved and might provide valuable proof for early diagnostic in diabetic nephropathy.

Objective To explore the serum levels changes of heart-type fatty acid-binding protein and brain natriuretic peptide in children with Kawasaki disease,as well as their links with the heart function. Methods A total of 53 children with Kawasaki disease admitted to our hospital from February 2008 to March 2012 were selected and at the same time 50 healthy controls were randomly selected. The serum levels for the children in acute phase,recovery period of Kawasaki disease and control group children were determined by en-zyme-linked immnoabsorbent assay. The internal diameter of coronary artery,LVEF,LVSF,CI and E/A of chil-dren were measured by two-dimensional echocardiography. Results The heart-type fatty acid-binding protein and brain natriuretic peptide concentrations of children with Kawasaki disease in acute stage[(18. 23 ± 13. 81) ng/ml,(517. 2 ±213. 2) pg/ml] were higher than those in recovery stage[(6. 29 ± 1. 26) ng/ml,(92. 1 ± 46. 0) pg/ml](P<0. 05). They were also higher for children in acute stage than those of control group[(6. 26 ± 1. 60) ng/ml,(37. 6 ± 7. 6) pg/ml]. The LVEF,LVSF,CI of Kawasaki disease for children in acute stage were lower than those in recovery phase[(60. 3 ± 3. 6)% vs. (64. 8 ± 4. 3)%,(30. 6 ± 2. 5)% vs. (34. 9 ± 1. 9)%,(3. 1 ± 0. 3)% vs. (3. 5 ± 0. 3)%]. Linear correlation analysis showed the heart-type fatty acid-binding protein and brain natriuretic peptide′concentrations in Kawasaki disease were negatively correlated with LVEF (P<0. 05). The heart-type fatty acid-binding protein′positive rates were also higher than brain natriuretic peptide for the children in acute and recovery phase. Conclusion The heart-type fatty acid-binding protein and brain na-triuretic peptide could be used as the biochemical markers for myocardial damage of children with Kawasaki dis-ease. The heart-type fatty acid-binding protein was more significant than brain natriuretic peptide. The heart-type fatty acid-binding protein and brain natriuretic peptide in Kawasaki disease were also high in recovery stage,indi-cating that myocardial damage could maintain for a long period after the clinical improvement.

Pulmonary hypertension is a group of diseases with high pressure of pulmonary circula-tion,severe cases can lead to increased right ventricular afterload,even right heart failure. Children pulmonary hypertension is difficult to detect in early stage because its nonspecific clinical manifestations. It is a serious complication of many respiratory diseases,it leads to decreased lung function and life quality. Screening early pulmonary hypertension of these children will facilitate it′s diagnosis,treatment and prognosis.

Objective:It is important to make sure hyperbaric oxygen chamber safe and stable for its generalized and high risk.Methods: The gas path, control system and operation of hyperbaric oxygen chamber should be maintained timing based on quality management and clinical using requirement.Results: Medical engineering technicians and clinical use of personnel to cooperate with each other, from the regular inspection, personnel management and preventive maintenance to make sure the safety use of hyperbaric oxygen chamber.Conclusion: Making relative system and emergence plan, strengthening train to users and strict management can make its used safe.

Objective:To develop a highly efficacious and sensitive immunological reagent for further investigation on the retinoic acid-induced gene I (Rig-I) of mouse .Methods:The Helicase domain coding region (726-2 240 bp) of mRig-I-H was cloned into plasmid pET15b (+) to construct the recombinant plasmid pET15b(+)-mRig-I-H.Then the plasmid was transformed into E.coli BL21 for protein expression.Rabbits were immunized with electrophoresis-purified recombinant protein to obtain the polyclonal antibody against mRig-I-H.The titer of polyclonal antibody was detected by ELISA and the specificity was identified by Western blot and Immunofluorescence.Results:The recombinant protein was expressed successfully in E.coli.Western blot analysis showed that target protein was expressed with a molecular weight of 40 kD.Titer of the polyclonal antibody was about 1∶1?105 by ELISA assay.With this antibody,we could detect the expression of Rig-I in RAW 264.7 cell line by Western blot and Immunofluorescence.Conclusion:The high level expression of Rig-I Helicase domain is induced in E.coli expressing system.Anti-mRig-I-H polyclonal antibody with high titer and fine specificity could be a novel tool in future investigation of Rig-I.

A case of a 47-year-old female patient who had a severe headache for 2 days was reported. Spontaneous subarachnoid hemorrhage (SAH, hunt-Hess classification Ⅱ level)-anterior communicating aneurysm rupture was diagnosed by brain CT and digital subtraction angiography (DSA) examination.Intravascular interventional aneurysm embolization was performed, and the patient appeared progressive limb weakness 5 days after operation. Physical examination revealed grade Ⅱ upper limb muscle strength and grade Ⅰ lower limb muscle strength. The muscle tone of the extremities was decreased,and the tendon reflex disappeared. The pinprick pain was decreased below the double wrist joints and the ankle joints. After neuroelectrophysiology test and cerebrospinal fluid assay, the patient was diagnosed as Guillain-Barré syndrome associated with subarachnoid hemorrhagic aneurysm embolization.

Objective To study the adenovirus infection and popular features in hospitalized respira-tory infection children of Qingdao area. Methods A total of 6 820 cases of respiratory infection from March 2011 to February 2014 in our hospital were enrolled,the average age was 5 years and 2 months old. Their ve-nous serums were collected on the first day of hospitalization,adenovirus IgM was monitored by indirect im-munofluorescence application. The adenovirus-positive rates of cases between different age groups, seasons and diseases were observed. Results Adenovirus were positive in 1 043 children,the rate was 15. 29%. The positive rate of young children group(1-3 years old) was 16. 92%,which was highest,there was statistic difference between young children group and other age groups(χ2 =12. 50,P<0. 05). Adenovirus infection rates were upward in recent years,the infection rates of winter and spring were higher than those of summer and fall,and there were significant differences between them(χ2 =28. 89,P <0. 05). The positive rate of AdV-IgM in children with severe respiratory tract infection was higher[19. 90%(877/4 408)]than that of the children with mild infection[12. 27%(296/2 412)](χ2 =64. 25,P<0. 05). Conclusion In recent years, adenovirus infection rate in Qingdao hospitalized respiratory infection children was rising,the infection rates in winter and spring were high,young children were the most vulnerable populations. The positive rate of ade-novirus infection in children with severe respiratory tract infection is higher than that of the children with mild respiratory tract infection.

The levels of IgG2b monoclonal antibodies against LD H-C4 in sera of mice bearing hybridoma backpack tumors secreting anti-LDH-C4-IgG2b were detected by quantitative ELISA． The accuracy between batches is 7.04%～l3.30 %, the intra-assay variation is 3.6l%～l0.20%． Standard curveof monoclonal lgG2b was well correlated (r=0.962　　884～0.996　　795)． The sensitivit y of the assay reach e dup to0.0l?mg/L． The present modified ELISA offers a reproducible, se nsitive, specific method in determination of antigen-specific IgG2b antibody in sera．

Objective To investigate clinical effect of in vitro maturation(IVM)of immature oocytes transferred from conventional in vitro fertilization embryo transfer(IVF-ET) cycles.Methods From January 2008 to June 2009.medical documents of 155 infertile patients underwent IVF-ET in the Reproductive Medical Center of Fimt Affiliated Hospital of Wenzhou Medical College were analyzed retrospectively.If more than 20 oocytes were monitored after 5-7 days of ovulation induction or follicular developmental retardation were confirmed after 8-13 days of ovulation induction.according to patients' wish,IVM were transferred in 60 cycles(group A).In the mean time.IVF was continued in 95 cycles (group B).The mean dosage of gonadotropin,the cancel lation rate of cycles,the mean numbers of oocytes retrieved and maturation,the rate of fertilization and excellent quality embryos.pregnancy outcome and the incidence rate of ovarian hyperstimulation syndrome(OHSS)were compared and analyzed.Results The rates of embryo transfer were 92%(55/60)in group A and 63%(60/95)in group B,which showed significant differences (P<0.05).In group A,the mean dosage of the gonadotropin,the mean number of oocytes retrieved,the cleavage rate and OHSS rate were(1030±468)U,10±6,82.2%(231/281)and O,and were(1544±338)U,14±4,94.0%(502/534)and 35%(21/60)in group B,respectively,all data abeve exhibited statistieal difference between two groups(P<0.05).However,the rates of fertilization and excellent quality embryos had no significant differences between two groups(P>0.05).In group A,the rate of clinical pregnancy per transfer was 53%(29/55)and multiple pregnancy was 14%(4/29),and were 47%(28/60) and 32%(9/28)in group B,they all had no significant differences (P>0.05).Conclusion IVM of immature oocytes used in conventional IVF cycles not only obtained a high clinical pregnancy rate,but also reduced gonadotropin using dosage and avoided OHSS completely.