The Wiskott-Aldrich syndrome(WAS)is a rare X-linked recessive immunodeficiency disorder characterized by thrombocytopenia and small platelets,eczema,recurrent infections,and increased risk for autoimmunity and malignancy.The gene responsible for this syndrome is WAS protein(WASP)gene.Six mutational hotspots have been identified now.Flow cytometric analysis of the WASP expression in lymphocytes is simple,rapid,and applicable for screening of WAS.WASP gene sequencing analysis can confirm the diagnosis.Hemato-poietic stem cell transplantation(HSCT)is the mainstay of treatment for WAS.So far more than 40 WAS patients have been diagnosed in China.Due to the few reports and varied clinical manifestations,WAS is often misdiagnosed as idiopathic thrombocytopenic purpura.This review will focus on recent progress in understanding the clinical presentations associated with mutations of the WASP gene,the genotype-phenotype correlation and management of this lethal disease.

0.05),but the markedly effective rate in the treatment group was superior to that in the control group(P

Acupuncture Points ; Aged ; Aged, 80 and over ; Bloodletting ; Female ; Humans ; Middle Aged ; Pruritus ; therapy

A simple and rapid expression and purification method of recombinant firefly luciferase was developed for bacteria detection. A modified luciferase gene from North American firefly Photinus pyralis was cloned into pET28a expression vector and the recombinant protein was produced in Escherichia coli BL21. The recombinant luciferase, equipped with a polyhistidine affinity tag, was purified by immobilized metal ion affinity chromatography (IMAC). The approach generated an abundant expression and an efficient purification of a recombinant luciferase with final yield 1.995mg/L of cell culture. Experiments on the recombinant luciferase also showed that the relative light units (RUL) of the enzyme were 5.8×108, and the specific activity was 2.9×1010 RLU/mg. By applying adenosine triphosphate (ATP) bioluminescence to detection of the coin bacteria using the recombinant protein, the ATP content of bacteria was 9.48×10-16mol/mL, and was identical to the bacteria counts (4500CFU/mL) in order of magnitude. Taken together, our results provided a simple and efficacious method of the preparation of recombinant luciferase, which could be applied in the determination of bacteria via ATP bioluminescence.

Animals ; Humans ; MicroRNAs ; Pneumonia ; genetics ; Pulmonary Fibrosis ; genetics

Objective To prepare monoclonal antibody(McAb) against human tissue kallikrein (HK) and develop an ELISA kit allows for the in vitro quantitative determination of human tissue kallikrein in urine. Methods To generate a monoclonal antibody specific for TK, the synthetic TK peptide consisting of 12 amine acids(12P), was fused to keyhole limpet hemocyanin(KLH) and used for immunization. Using hybridoma screening, monoclonal secreting cell lines were identified and used to generate ascites in BALB/c mouse. Antibody was purified by affinity column chromatography. 12% SDS-PAGE and Western blot were used to visualize the purified antibody. This kit employs indirect competitive ELISA technique and BiotinAvidin System. 12P was fused to bovine serum albumin(BSA) and has been pre-coated onto a microplate at first. Standards and samples were added to the appropriate microplate wells with a biotin-conjugated McAb croplate well. A TMB substrate solution is added to each well. The enzyme-substrate reaction is terminating by the addition of a sulphuric acid solution and the color change is measured spectrotometrically at a wavelength of 450 nm. The concentration of tissue kallikrein in the samples is then determined by comparing the O.D. of the samples to the standard curve. Results 8 hybridoma cell lines secreting mAbs special to HK,SDS-PAGE and Western blot demonstrated successful preparing and purification of McAb( 100% ). The linearity of this ELISA kit is demonstrated(r =0. 990). The range of detection of the assay is 0.008 μg/ml to 0. 5 μg/ml. The assay remained stable, with no change in the values measured, over five cycles of freezing and thawing. Conclusion 8 McAbs against HK have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for detecting HK can meet the needs of detection of HK in urine samples.

OBJECTIVETo explore the effects of chronic hypoxia on left and right ventricular function and the expression of cardiac transient receptor potential canonical (TRPC) channels in rats.

METHODSForty eight SD male rats were randomly divided into control group (CON) and chronic hypoxic pulmonary hypertension model group (CH) (n = 24). In CH group, rats were exposed in chronic hypoxia environment (10% +/- 0.2% O2) to induce myocardial hypertrophy. After 3 weeks, mean systemic arterial pressure (mSAP), right ventricular systolic pressure (RVSP), left ventricular systolic pressure (LVSP), left or right ventricular pressure maximum rate of rise (LV/RV + dp/dt(max)), left or right ventricular pressure maximum rate of descent (LV/RV-dp/dt(max)), right ventricular hypertrophy index (RVMI) and left ventricular hypertrophy index (LVMI) were measured. Left and right ventricular myocardium tissue sections were stained by HE and observed under light microscope. Real-time polymerase chain reaction (real-time-PCR) and Western blot were performed to detect the expression of TRPC subfamily.

RESULTSRVSP, RVMI, RV + dp/dt(max) and RV-dp/dt(max) were markedly elevated in CH group (P < 0.01) in comparison to CON group. LVMI was markedly reduced in CH group in comparison to CON group (P < 0.01). LVSP, LV + dp/dt(max) and LV- dp/dt(max) had no significant changes in CH group in comparison to CON group. Right ventricular myocardial cells of CH group became thick, the nuclei stained deeply, the shape of nuclei became not regularity. Left ventricular myocardial fibers did not change significantly. There was significant difference in the levels of mRNA and protein of TRPC1 between CON and CH groups.

CONCLUSIONFor three weeks exposed to chronic hypoxia induced right ventricular hypertrophy specifically, raised the mRNA and protein expression of TRPC1 on right ventricular myocardial cells . TRPC1 might be involved in the development of cardiac hypertrophy.

Animals ; Disease Models, Animal ; Hypertension, Pulmonary ; metabolism ; physiopathology ; Hypoxia ; metabolism ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley ; Transient Receptor Potential Channels ; metabolism ; Ventricular Function, Left ; physiology ; Ventricular Function, Right ; physiology

Objective To study the effect of intratracheal anti-tumor necrosis factor-α antibody(TNF-αAb)on ultra-structure of lung after cardiopulmonary bypass(CPB).Method 28 healthy rabbits were selected and randomly evenly divided into four groups:I group only received open chest operation;Ⅱ-Ⅳ groups underwent CPB.In the IV group,rabbit TNF-α Ab (2400 ps/kg)Was dropped into the intracheal tube before operation and just after releasing the aortic clamp.Saline was given to the Ⅲ group by the same way.Water volume,TNF-α mRNA,TNF-α protein,apoptosis and pathomorphological changes were measured in the lung tissues.Results TNF-α Ab can re-duce releasing of TNF-α.It could also reduce the occurrence of apeptosis and attenuate pathomorphological changes in the lung tissue.Conclusion Intratracheal TNF-α Ab markedly lessenes the injury of nltrastructure of lung after CPB.

This study aims to investigate the relationship between odor and contents of the chemical compounds in Lonicera japonica, including chlorogenic acid, galuteolin and polyphenols. High performance liquid chromatography (HPLC) was applied to determine the contents of chlorogenic acid and galuteolin in L. japonica. The ponptent of polyphenols was determined by UV-Vis Spectrophotometry. Electronic nose was used to extract and measure the odor of L. japonica. Then SPSS 17.0 software was employed for data processing. There is a significant positive correlation between the comprehensive index value of aroma and the contents of chlorogenic acid and polyphenols. The regression equations have been established. However, the relationship between the comprehensive index value and the content of galuteolin is not obvious. This is proof that the odor of L. japonica has close connection with the chemical compounds. Therefore, this research offered a new method for initially determine or predict the content of the chemical composition in L. japonica,
Chlorogenic Acid ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Electronic Nose ; Lonicera ; chemistry ; Odorants ; analysis ; Polyphenols ; chemistry ; Smell

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.