Animals ; Electrochemical Techniques ; methods ; Gene Silencing ; Genes, Reporter ; Humans ; MicroRNAs ; blood ; genetics ; metabolism ; physiology ; Microarray Analysis ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

The specific ScFv gene against botulinum neurotoxin A (BoNTa)was cloned into pPIC9k. Positive integrators were screened by increasing the dose of G418 in culture and expressed in Pichia pastoris GS115. As a result, engineered recombinant clone were obtained. 26 kD product of interest was seen easily in SDS-PAGE. Expression of human ScFv got the highest level 15% of total secreted proteins during 72～84 h after 1% methanol inducing. Purification of ScFv was finished by two steps: gel filter and ion exchange. Competing ELISA showed that recombinant ScFv could compete with antiserum to specific bind BoNTa.

ObjectiveTo study the differentiation of adipose tissue-derived mesenchymal stem cells (ADMSCs) into cardiomyocytes in vitro. MethodsADMSCs were isolated and purified by the method of digesting and adhering to the culture plastis. The third generation of cells were determined with immunocytochemistry method and induced by 5-aza. On the 7th, 14th, 21st, 28th day after being induced, the surface antigen of myoblast cell and cardiac myocyte were determined, the expression of gene of GATA4 and Nkx2.5 were tested, and the content of atrial natriuretic polypeptide (ANP) assayed. ResultsImmunocytochemical staining showed CD44, CD13, CD105 positive, CD45, CD34, HLA-DR, factor Ⅷ negative. After being induced by 5-aza, the direction of cell arraying was gradually similar. The cells were stained positively for Desmin, α-Sarcomeric Actin, myosin heavy chain and Troponin T. The cells express GATA4 and Nkx2.5 and secrete ANP. ConclusionThere was mesenchymal stem cells in human adipose tissue. 5-aza may induce ADMSCs to differentiate into cardiomyocytes in vitro.

Objective To investigate the clinical features,treatment response and prognosis in children with Takayasu′s arteritis(TA) in order to improve the understanding of TA.Methods A retrospective study of 10 children with TA was performed.All of them were admitted and diagnosed in Peking University First Hospital from Jan.1998 to Oct.2008.The clinical features,laboratory tests,imaging modalities,treatment response and prognosis were all collected and evaluated.Results There were 3 boys and 7 girls in the 10 patients with TA,and the ratio of male to female was 12.3.The onset was from 4 months to 9 years old,with average age at 5.5 years old.The average duration of diagnosis was 7.6 months.The incidences of hypertension,vascular bruits,albuminuria,convulsion were present in 100%,100%,70% and 40%,respectively.The clinical types included typeⅡ(60%),type Ⅲ(10%) and type Ⅳ(30%).The acute phase inflammatory indices of activity such as erythrocyte sedimentation rate(ESR),C-reactive protein(CRP) and white blood cell(WBC) were not evidently increased.Tuberculosis infection was found in 6 out of 10 patients and anti-tuberculosis treatment was performed.Six patients were treated with steroids and 3 cases of them were also given immunosuppressives cyclophosphamide or methotrexate.Three of the 10 patients received anti-hypertensive and vasodilator.Two patients received percutaneous translurminal angioplasty and 1 patient received nephrectomy.One patient died of renal failure,heart failure and shock.Conclusions The patients with TA had high prevalence of tuberculosis infection,diagnosis as often late because of lack of specific clinical features at the acute inflammatory period.When organic ischaemia occurred,treatment response was usually unsatisfactory.Patients with multi-systemic and multi-viscera lesions should have comprehensive examination,especially for those with hypertension,pulseless and vascular bruits,in order to rule out TA.Early ultrasonography,computed tomography and magnetic resonnance image methods are valued in eariler diagnosis and they are the key factors to improve prognosis.

China ; Female ; Gynecology ; trends ; Humans ; Physical Examination ; statistics & numerical data ; Rural Health ; statistics & numerical data ; Rural Population

1:16) of the relative serum antibody has been observed in 80%, while none of this high level was observed in patients with Echovirus-25 and adenovirus dermatitis (

Objective To investigate the proliferation of rabbit adipose-derived stem cell(ADSCs) transfected by adenoviral vector mediated hTGF-?_1 gene and its chondrocyte differentiation potential.Methods The Ad-hTGF-?_1 plasmid vetor which had the hTGF-?_1 gene was developed and transfected the ADSCs.The experimental group was the hTGF-?_1 transfected group.The cells enclosed by alginate were cultured in com- plete chondrogenie medium(CMM).The morphology of the cells were observed,and RT-PCR was used to measure hTGF-?_1 and collagenⅡexpression,at the same time western blot and immunohistochemistry were applied to detect the expression of collagenⅡin ADSCs before and after transfected with hTGF-?_1 gene. Results The hTGF-?_1 transfected ADSCs became the polygon and it proliferated well.The RT-PCR result of hTGF-?_1 on the transfected group was better than the control after transtected for 7 day and 21 day.The dif- ference between the two groups was significant(P

In order to find an easy and rapid quantitative analytical method to detect rhamnolipid produced by Pseudomonas aeruginosa, three methods, H_ 2 SO_ 4 -anthrone analysis method, L-cysteine-H_ 2 SO_ 4 method and phenol-H_ 2 SO_ 4 method, were compared in the present paper, and the influence factors were also considered.The results showed that H_ 2 SO_ 4 -Anthrone analysis method was better than the others and its optimal reaction condition was obtained.The influence to the quantitative analysis of rhamnolipid from the residual glucose and the top clean liquid layer in the ferment solution could be ignored.But the influence from the bacterial body and the middle layer of the ferment solution reached a certain degree.Thus, the bacterial body should be removed before measuring.However, the influence from the middle layer of the ferment solution could be avoided by making a standard curve which was made by using a rhamnose mixed with the middle layer ferment solution.

Objective To study the different expressions of receptor activator of nuclear factor-kappa B ligang(RANKL) mRNA in spleens of rats fed with diet of low calcium and high fluoride. Methods A 2× 2×2 factorial design was used and the factors were calcium, fluoride and action time. In the design, 40 Wistar rats [average body mass(118.9±13.5)g] were divided into four groups randomly by weight: control with normal diet (0.790%, calcium), low calcium group with low calcium intake(0.063%, calcium), high fluoride group with normal diet and high fluoride intake(100 mg/L, fluoride) and low calcium and high fluoride group with low calcium and high fluoride intake. After 4 and 8 months, 5 rats of each group were sacrificed and total RNA was extracted from spleen. And the expression levels of RANKL mRNA were determined by reverse transcription polymerase chain reaction (RT-PCR). Results At time of 4 months, the expression level of RANKL mRNA was 0.13± 0.05,0.13± 0.03,0.17±0.02,0.27± 0.05 and at time of 8 months, it was 0.11 ± 0.01,0.16 ± 0.02,0.16± 0.03,0.36 ± 0.07 in control group, low calcium group, high fluoride group, low calcium with high fluoride group, repectively. The factorial design AVONA showed that low calcium and high fluoride had significant effects on RANKL mRNA expression(F = 40.224,56.679, all P < 0.05) while action time had not(F = 2.850, P > 0.05 ). The interactions of low calcium with high fluoride or high fluoride with action time were signifieant(F = 7.247, 18.789, all P < 0.05) while the interaction of high fluoride with action time was not(F = 1.751, P > 0.05). Conclusions Low calcium alone or high fluoride alone or low calcium with high fluoride or low calcium with action time can increase the the RANKL mRNA expression level. High fluoride does not affect the RANKL mRNA level as the action time is prolonged.

Objective To explore the prognostic value of pediatric critical illness score(PCIS)and pediatric risk of mortality score(PRISMⅢ)and the accuracy for evaluating the state of children with acute respiratory distress syndrome(ARDS).Methods Seventy-one cases hospitalized children from 29 days to 14 years old of Hebei ARDS cooperation group were selected during the 13 months between 2005 and 2006.All cases were confirmed according to ARDS diagnostic standard.For prospective studies,the patients were scored simultaneously with PCIS and PRISMⅢ at different times:when the patients entered PICU,when the patients were in the worst situation in PICU,when the patients were diagnosed as ARDS and when ARDS was serious.The data were performed by using Logistic regression etc.Results Values of Logistic regression were P