Humans ; Vision Disorders/etiology* ; Visual Acuity

Objective To clone the gene of arginine kinase (AK) from Periplaneta americana, produce its recombinant protein and investigate its allergenicity. Method The cDNA of AK was cloned using specific primers from the total RNA of P. americana. The cloned gene was inserted into pMD18-T vector and digested by BamHI and HindⅢ. The cDNA was sequenced and subcloned into pET-28a expression vector. The cloned AK cDNA gene was expressed in Escherichia coli BL21 (DE3) by IPTG induction. The recombinant AK (rAK) was purified by metal (Ni2+) chelating affinity chromatography. Its allergenicity was examined by both Western blotting and enzyme-linked immunosorbent assay (ELISA). Result The cloned cDNA ORF sequence (Accession no. EU429466) contained 1 068 bp and encoded 365 amino acids. Its sequence homology with the published one (Accession no. AY563004) was 99.9% at nucleotide level. The allergen rAK was highly expressed in E. coli BL21 (DE3) as a soluble protein mainly with the molecular weight of about Mr 45 000 under induction of IPTG and purified by 6-His-tag purification system. Both in the non-denaturalization and denaturalization conditions, the recombinant allergen was identified as its affinity to IgE antibodies from the cockroach-allergic patient sera by Western blotting and ELISA. Conclusion The recombinant cockroach arginine kinase has been obtained with proper allergenicity.

Objective To establish a novel real-time PCR method to detect HCV RNA using Selfreporting duplex mutation primers.Methods The recombinant vector pMD18-T-HCV 5′-NCR was used as the calibrator.The Self-reporting duplex mutation primers were designed according to the gene sequence.And then the PCR reaction system was optimized and evaluated.The specificity,sensitivity and reproducibility of real-time PCR were estimated,The serum specimens from 90 cases(30 cases of HCV,30 cases of other viral hepatitis and 30 healthy volunteers) were tested with this real-time PCR; Results were compared with those obtained using a commercial TaqMan kit.Results The assay was established.It showed linearity over a wide range from 20 - 109 IU/ml.Intra-experimental coefficients of variation(CVs) were 1.37% -4.59%,and inter-experimental CVs were 1.58% -4.81%,respectively.There was no significant difference of HCV genome number tested by the two methods(R2 = 0.95) in 30 hepatitis C patients; HCV DNA was not detected in any serum samples of 30 healthy volunteers by the two methods.The specificity was 100%(60/60).All the samples in patients with clinically confirmed HCV infections showed HCV RNA positive.There wass good correlation between the quantitaive results and results obtained using the commercial TaqMan kit.Conclusions It is demonstrated that real-time PCR is a reliable,accurate and feasible assay for HCV.The establishment of this assay provided alternative technology for clinical diagnosis or therapeutic drug monitoring in the field of HCV infection and epidemiologic survey.

Air Pollutants, Occupational ; analysis ; Guanidines ; analysis ; Humans ; Occupational Exposure ; analysis ; Threshold Limit Values ; Workplace

Adolescent ; Dermatitis Herpetiformis ; complications ; Esophageal Diseases ; etiology ; Humans ; Male ; Mucous Membrane ; pathology

Animals ; Animals, Newborn ; Hippocampus ; cytology ; drug effects ; physiology ; Interleukin-6 ; pharmacology ; Ion Channels ; drug effects ; physiology ; N-Methylaspartate ; physiology ; Neurons ; drug effects ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Wistar

Animals ; Caudate Nucleus ; drug effects ; physiology ; Dopamine Antagonists ; pharmacology ; Rats ; Rats, Wistar ; Substantia Nigra ; drug effects ; physiology

Objective To investigate the effects of montelukast(MK),a selective cysteinyl leukotriene receptor 1 antagonist on interleukin(IL)-5 and eotaxin expression as well as serum total immunoglobulin E(IgE) production during MK treatment of mouse acute asthma airway inflammation.Methods Sensitized Balb/c mice were challenged by ovabumin(OVA)to establish the acute asthmatic mo-(del).Twenty-five mg/kg of MK(MK group) or Saline(Saline group)were given by intravenously for 3 days.Cellular infiltration of bronchoalveolar larvage fluid(BALF) were assessed by Wright staining.Production of IL-5 and eotaxin in the lung or BALF and serum IgE were determined by enzyme linked immunosorbent assay(ELISA).The expression of IL-5 and eotaxin mRNA were measured by semi-quantified reverse transcriptase-polymerase chain reaction(RT-PCR).Plasma MK level was determined by liquid chromatography.Results After 24 h OVA challenge,the numbers of total white blood cells,neutrophils and eosinophils(EOS)of BALF were(26.0?18.9)?10~7/L,(5.92?8.09)?10~7/L and(0.74?0.88)?10~7/L in MK treatment group.They were significantly reduced compared with those in Saline group,respectively(P80%.The level of IL-5 in BALF and lung tissue were(48.52?14.45) ng/L and(27.40?9.62) ng/g protein in MK group,which significantly declined compared with that in saline group;BALF eotaxin level declined too.Serum IL-5 and total IgE level were also significantly reduced;IL-5 mRNA expression in lung significantly decreased.Eotaxin and its mRNA expression in lung were not decreased significantly.Conclusion MK(exerts) its anti-inflammatory effect mainly through the suppression of IL-5 and IgE production.

Objective To summarize the clinical characteristics and causes of ischemic cerebrovascular disease(ICD)in children.Methods A retrospective analysis was conducted on the clinical data of 53 cases with ICD from Feb.2002 to Jun.2008 at the department of neurology in Wuhan Children's Hospital.The self-designed questionnaire of children with ICD was used,whose items included patients' age,gender,personal history,clinical features,cerebrospinal fluid examination,neurological imaging,immunologic examination,metabolic examination,and so on.Results Of 53 children with ICD,30 cases(56.6%)were male,and 23 cases(43.4%)were female.Patients' age varied from 9 months to 12 years old,in which 45 cases(84.9%)were less than 6 years old.Patients from rural area(60.4%)were more than those from city(39.6%).Ratio of limb paralysis was 75.5%(40 cases)in first clinical symptomatology of children with ICD,including hemiplegia in 32 cases(60.4%),alternate hemiplegia in 5 cases(9.4%)and monoplegia in 3 cases(5.7%).Skull CT/MRI scan was performed to reveal 27 cases(50.9%)with basal ganglia region infarction and secondly 15 cases(28.3%)with multi-lobar infarction.Forty cases were found in abnormal cerebrovascular image by means of magnetic resonance angiography/digital subtraction angiography,in which middle cerebral artery and its branches were involved in 21 cases(52.5%).There were 41 cases(77.4%)of patients to be found with clear causes,of which 13 cases(24.5%)were of infections,8 cases(15.1%)of moyamoya disease,5 cases(9.4%)of cerebral vascular malformations,4 cases(7.5%)of head trauma.However,another 12 cases(22.6%)of patients had unknown etiology.Conclusions Children with ICD had characteristics themselves.The limb paralysis was mostly the first symptoms,and the middle cerebral artery and its branches lesions were the most common locations in children with ICD,and next the internal carotid artery involvement,anterior cerebral artery involvement,posterior cerebral artery involvement,cerebral vascular malformations,and so on.Their major cause was infection,followed by Moyamoya disease,cerebrovascular malformations and head trauma,and there were still some unknown causes.

Objective: To explore the cause and the strategy for air quantity signaling in old patients using mechanic air. Method: Summary and analysis of the causes for air quantity signaling in 187 cases of old patients using respiratory machine. Results: One hundred fifty-one cases showed low-limited air quantity signaling per minute whereas 36 cases presented high-limited air quantity signaling per minute. Conclusion: Judging and removing the obstacle in respiratory machine on time and accurately are the key point raising both the survival rate of severe patients and the successful rate of mechanic air.