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Objective To analyze the immunological characteristics of Trichinella spiralis secretory antigen P53 and to evaluate its value in diagnosis of trichinellosis. Methods An open read frame of secretory antigen P53 was cloned from Trichinella spiralis by reverse transcriptasepolymerase chain reaction (RT-PCR) and then sequenced. Bioinformatics analysis was performed to search for its homologues in other helminths and predict its potential linear B cell epitopes and T cell epitopes. The sequence coding mature peptide was inserted into prokaryotic expression vector pET28a(+) and the purified recombinant product was identified by Western blot using serum samples of patients infected with Trichinella spiralis or other helminth. Results Bioinformaties analysis results showed that there was no P53 homologue in other helminths, which indicated that there were many linear B cell epitopes and T cell epitopes in TsP53. The recombinant P53 antigen only reacted with the serum samples of patients infected with Trichinella spiralis without any cross-reaction with the serum of patients infected with other helminths. Conclusion P53 has strong immunogenicity and immunoreactivity, which may be a promising candidate for developing Trichinella spiralis specific diagnostic method.
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Objective: To discovery the central mechanism of acupuncture Heart Meridian precondition myocardial ischemia in gene expression pattern, the authors applied gene chip tech to filter variably expressed genes in hypothalamus. Methods: Rats were seperated into normal, model, acupuncture Heart Meridian group and acup Lung Meridian group randomly. Acute myocardial ischemia rat model was made with ligation left anterior descending branch of the coronary artery. After model succeed, select hypothalamus seperately and mixed the same group together of 3 rats. Then applied Rats U230A genechip refered by Affymetrix Co. to compare the variations between these groups. Results: To compare with normal group, differential expression genes and expression sequence tags (ESTs) in model group were 73 with signal log ratio ≤ 1 and 92 with signal log ratio ≤-1 , mainly included ion channel, calcium/ calmodulin-dependent protein kinase Ⅱ inhibitor, antigen and so on. Similarly, compared with model group, differential expression genes and expression sequence tags (ESTs) were 190 with signal log ratio ≥ 1 and 34 with signal log ratio ≤-1 in acupuncture Heart Meridian group, mainly included 5-hydroxytryptamine receptor, cellular metabolism, fatty, immuno reaction, G-protein coupled receptors, ion transport, signal transductions and so on, while in acupucnture Lung Meridian, the number is 57 and 26 correspondly. Conclusion: There were exactly variations in hypothalamus mechanism that relate to acupuncture Heart and Lung Meridians.
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Objective To detect human metapneumovirus (hMPV)in respiratory intection rapidly and perform molecular analysis of hMPV.Methods Seven respiratory tract virus(11 subtypes)were assessed using multiplex PCR technology and flexible Multi-Analyte Profiling(suspension array).Human metapneumovirus was confirmed by using a real.Time reverse ranscriptase CR(RT-PCR)assay followed by sequencing.The cladogram analysis was performed further.Results The virus were detected in 40.2%(19/47)samples collected from clinicsl respiratory tract infections,including 8(42.1%)HRSV,7(36.8%)influenza virus,1(5.3%)parainfluenza virus,1(5.3%)rhinovirus,1(5.3%) coxsackievirus and 1(5.3%)human etapneumovirus infections.This is the first time that hMPV was deteced from clinical samples in Shenzhen.The sequencing of specific fragment of neucleoprotein of hMPV showed this hMPV shares over 98% homology with Beijing strain.Japan strain and Thailand strain.The cladogram analysis showed that they were in the same cluste.Conclusions Human etapneumovirus is a maior cause of children respiratory tract disease. Multiplex PCR technology and nexible Multi-Analyte Profiling were hish sensitive and high-throughput for detection of human metapneumovirus.They axe very robust and applicable in etiology analysis.
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Objective To analyze the expression of apoptosis-relared genes of retinal blood vessel in early diabetic rats by gene chip technology. Methods To make diabetic rat model by intraperitoneal injection of streptozotocin (STZ). On the 6th week after blood pressure increased, 10 rats were executed in Diabetic group and normal control group respectively. 20 retinal blood vessels were extracted and the RNA was isolated. The probe was made of α-32 P-deoxyadenosine triphosphate (dATP)-labeled sample which hybridized 1176 nylon chips, and then analyzed by software. Three different expression genes were selected to verify by reverse transcription polymerase chain reaction (RT-PCR). Results On the 6th week, 136 (11.5%)genes were differentially expressed [up-regulated genes were 90 (7.6%), down- regulated genes were 46(3.9%)] in diabetic group. These genes involved into different groups according to their function. Especially in 72 apoprosis-related genes, 15 genes were differentially expressed. The up- regulated genes were some TNF receptor family members such as TNFRSF12, TRAIL, TNFRSF9, FADD;Bcl-2 family members such as bcl-w, bax, bakl and AKT. The down-regulated genes were FAF1 which related to fas. Conclusions The expression of retinal vascular gene in early diabetic rats has been changed complicatedly. In particular, the multiple apoptosis-related genes have been changed in early diabetic, and most of them are at the upstream of apoptosis pathway. These findings indicate that the development of diabetic retinopathy is associated with multiple signaling pathways leading to apoptosis, while the alterations on the level of molecular biochemistry are still limited in apoptosis induction period.