Objective To study the immunodiagnostic potential of recombinant membrane protein Tetraspanin 2 of Schistosoma japonicum(rSjTsp2) as a diagnostic antigen. Methods The rSjTsp2 was expressed and purified as an antigen used in indirect ELISA to detect specific serum antibody against Schistosoma japonicum of rabbits that at different stages of infection, and the results were compared with those of ELISA using natural antigen (AWA) as antigen. Results The sensitivities of rSjTsp2-ELISA were 75.4% (46/61),77.6% (45/58) and 81.5% (44/54), respectively, 2, 4, 6 weeks after rabbits were infected with Schistosoma japonicum, meanwhile the sensitivities of AWA-ELISA were 68.9% (42/61),86.2% (50/58) and 88.9% (48/54), respectively. The sensitivity difference of two methods was not significant statistically (x2 = 0.652, 1.454,1.174, all P ＞ 0.05).Conclusions The rSjTsp2 is expressed successfully in vitro, and rSjTsp2 recombinant protein expressed in prokaryotic system has a certain application potential in immunodiagnosis.

Objective To observe the clinical effects of hyperbaric oxygen combined large dose ganglioside on delayed encephalopathy after acute carbon monoxide poisoning(DEACMP).Methods A totol of 72 patients of DEACMP were randomly divided into the observation group(37)and the control group(35).The patients of control group were given the routine treatment with hyperbaric oxygen.In the treatment group patients were given large doses of gangliosides 100 mg,1 times a day,for a period of 20 to 30 days additionally to the routine treatment of hyperbaric oxygen.All patients were examined before and after treatment in EEG examination (EEG),mini-mental state examination(MMSE).Results The abnormal rate of EEG was 37.83％(14/37) in the control group,which was significantly lower than that of 65.71％(23/35) in the treatment group((x2 =5.60,P ＜ 0.05).In the treatment group,the MMSE was(15.45±2.93) and(23.70±2.13) before and after treatment,respectively; in the control group,the MMSE was(14.88±2.84) and(20.33±2.09) before and after treatment.The MMSE was significantly impoved after treatment in both group(t =3.18 and 2.91,Ps ＜0.05).Furthermore,the MMSE impoved more significantly in the treatment group compared to the control group (t =6.28,P ＜ 0.05).The effective rate of treatment group was 86.48％(32/37),which was significantly higher than that of 51.43％(20/35) in the control group(x2 =7.72,P ＜ 0.01).Conclusion Application of large dose ganglioside treatment had significant clinical effects in delayed encephalopathy after acute car-bon monoxide poisoning,which is better than the effect of routine hyperbaric oxygen treatment.

Objective To express Schistosoma japonicum Mago nashi(SjMago)gene,and prepare its specific polyclonal antibody.Methods SjMago gene was amplified by PCR from Schistosomulum cDNA library and subcloned into pET28a(+)vector,its recombinant proteins were expressed with IPTG.Rabbits were immunized with the polyacrylamide gel particles containing the recombinant proteins for polyclonal antibody preparation,the sera were detected for antibody specificity by Western blot and titer by ELISA assay.Results SjMago prokaryotic expression plasmid was successfully recombined and the target proteins was induced by IPTG in a molecular weight of 17 X 103，the high titer(1∶40 960)polyclonal antibody was isolated from the immunized rabbit，specific rotein band was detected by Western blot.Conclusion SjMago protein has been successfully expressed and its specific polyantibody is prepared,which lays the foundation for further study.

Objective To investigate the changes and pathogenic significance of serum interleukin-12p70 (IL-12), intefferon-γ,(IFN-γ) and IL-4 in the course of hemorrhagic fever with renal syndrome(HFRS). Methods Twenty five eases were divided into mild group (14 eases) and severe group (11 cases) according to the severity of illness. Blood samples were collected in various stages(fever, hypotensian and oliguria,diuresis stage). Serum IL-12 and IFN-γ levels were determined by enzyme-linked immunoserbent assay(ELISA), IL-4 by radioimmunoassay (RIA), blood urea nitrogen (BUN) and platelet by automatic biochemical analyzer and blood analyzer. Results Serum IL-12 levels in mild and severe groups were significantly different during various stages of HFRS (F=5.765, P<0.01). The IL-12 level of both patient groups significantly increased(P<0.01) in fever[ (0.87±0.38), (1.08± 0.77)μg/L], hypotension and oliguria [ (0.77±0.21), (2.11±2.13)μg/L] ,and diuresis stage [ (1.42±1.10), (1.20±0.88)μg/L], compared with control group [(0.56±0.10)μg/L]. In various stages, IFN-γ levels of both case groups were respectively (8.04±13.05), (5.94±8.24), (15.95±18.05), (4.41±4.10), (1.09±1.24), (1.38±1.74), (1.12±1.26), (0.19±1.29)μg/L, and the difference was statistically significant compared with control [ (0.27±0.15)rig/L]. K,-4 levels did not change significantly in the stages(F=0.682, P0.05), while the ratios of IFN-γ and IL-4 contents in mild and severe cases were significantly higher than control [(0.36±0.26) μg/L] in fever[ (2.46±3.52), (16.92±22.77)p.g/L], hypotension and oliguria[(2.52±2.72), (1.77±2.06) μg/L],diuresis stage [(1.45±2.28), (2.32±3.98)μg/L], the difference had statically significant (P<0.05 or 0.01).The curve of IL-12 was similar to that of BUN, but was contrary to blood platelet count. Conclusions The elevated levels of IL-12 and IFN-γ, with the imbalance of Th1/Th2 might be the main cause of systemic inflammatoryresponse and involved in the pathogenesis of HFRS.

To investigate the epidemic and evolutionary trends of enterovirus (EV) in the external environment of Yunnan Province, China, molecular typing was performed on 4 EV strains that were isolated from environmental sewage in Yunnan. The VP1 region of isolates was amplified by RT-PCR using universal enterovirus primers, and the amplified VP1 region was sequenced for GenBank BLAST search and genotype analysis. The 4 EV strains were identified as ECHO7. Their nucleotide and amino acid homologies with the VP1 sequences of 68 ECHO7 strains retrieved from GenBank were measured by Mega software analysis. Our findings showed that ECHO7 strains from environmental sewage and population samples were in different evolutionary branches. These strains showed typical geographical and temporal differences; In addition, there were different transmission chains at the same time and in the same area. ECHO7 strains isolated from sewage water and patients with acute flaccid paralysis during the same period in Yunnan belonged to different clusters and evolved at different speeds. Special concerns are needed for this problem. Continuous molecular biological surveillance of human EV in the external environment of Yunnan will provide strong support for early warning of EV diseases.
China ; Databases, Genetic ; Enterovirus ; genetics ; isolation & purification ; Evolution, Molecular ; Humans ; Molecular Epidemiology ; Sequence Analysis ; Sewage ; virology

The genetic diversity of three Tibetan herbs, i. e., Sang-Di, E-Dewa and Ye-Xingba (Tibetan names), was studied based on the field collection, specimen identification and DNA sequence analysis. Swertia hispidicalyx, Gentiana lhassica and Scrophularia dentata, as the original plants of the three Tibetan herbs, were collected and identified. The regions of ITS, matK, rbcL, rpoC1, trnL(UAA), psbA-trnH, atpB-rbcL, trnS (GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF(GAA) and nadl 2nd intron were amplified and sequenced. The ITS regions of S. hispidicalyx and S. dentata were cloned and sequenced, and the sequences were classified into different genotypes. All the sequences were analyzed and compared with those of closely related species. Our studies may provide reference for the genetic diversity analysis and molecular identification of the three Tibetan herbs.
Genetic Variation ; Gentiana ; classification ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; classification ; genetics ; Scrophularia ; classification ; genetics ; Swertia ; classification ; genetics ; Tibet

This study aims to establish a new method for quality evaluation of Gentianae Macrophyllae Radix by simultaneous determination of five iridoids (loganic acid, 6'-O-beta-D-glucopyranosylgentiopicroside, swertiamarin, gentiopicroside, sweroside), and to detect five iridoids in the root of eight species (Gentiana macrophylla, G. straminea, G. crassicaulis, G. dahurica, G. robusta, G. waltonii, G. lhassica, and G. tibetica). The separation was carried out on a Shiseido SPOLAR C18 (4.6 mm x 250 mm, 5 microm) column eluted with mobile phase of water containing 0.04% formic acid (A) and acetonitrile (B) in a gradient program. The flow rate was 0.8 mL x min(-1). The detect wavelength was set at 240 nm. The column temperature was kept at 30 degrees C. The volume of injection was 5 microL. The five iridoids were well separated with ideal linear correlations. The average recoveries were 97.35% - 106.23%. All the five iridoids were detected in the root of eight species. The contents of same species changed in a somewhat wider range. The contents in root of G. dahurica were lower than that in other species.
China ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Gentianella ; chemistry ; Iridoid Glycosides ; analysis

Aminoacyl-tRNA syntheses (AARS) can catalyze the adenosine triphosphate (ATP)-dependent acylation of their cognate tRNA(s) with a specific amino acid. They can be seen as an index to reflect the energy metabolic rate of ischemic brain cells in ischemic penumbra. This study examined the relationship between arginyl-tRNA synthetase (ArgRS), one of the AARS, and cerebral ischemia in rats. The model of middle cerebral artery occlusion (MCAO) was established in rats. The expression levels of ArgRS protein and mRNA were detected in rat brain tissues at different time points following MCAO by Western blotting and RT-PCR, respectively. The results showed that the MCAO model was successfully established. Western blotting and RT-PCR analysis revealed that the ArgRS protein and mRNA were expressed in brain cells in both ischemic and normal penumbra tissues. The expression levels of ArgRS protein and mRNA peaked at 6 h after MCAO and decreased gradually. At 24 h, the expression levels of ArgRs protein and mRNA in ischemic penumbral tissues were lower than those in normal tissues. The expression levels of ArgRS mRNA and protein in ischemic penumbra varied with ischemic time, suggesting that the energy metabolism of brain cells in penumbra changed dynamically after ischemia to ensure the endogenous self-protection of the body. The brain oxygen supply should be improved as soon as possible, especially within 6-12 h after ischemia, so as to meet the demand for energy metabolism in ischemic penumbra and make sure the cell structure remains stable.

Objective：To study the clinical and pathologic features of patients with lupus nephritis (LN) whose myeloperoxidase anti-neutrophil cytoplasmic antibody (MPO-ANCA) were positive. Methods：The clinical and pathological features were analyzed in 18 patients with LN whose MPO-ANCA were positive. And the data of patients with different clinical outcomes were compared. Results：（1）The hematological abnormalities, hypertension and serositis in these patients were more common than general ones with LN. （2）Proteinuria and hematuria were common, the morbidities of gross hematuria and renal failure in these patients were higher than general ones with LN.（3）Various autoantibodies were positive in these patients.（4）Segmental necrosis crescentic nephritis accompanied by density of immunocomplex in glomeruli and vasculitis in intestitium were common.（5）The morbidity of ESRF and mortality of these patients were similar to general ones with LN. The morbidity of tubular atrophy in those with poor prognosis was significantly higher than those survived. Conclusion：The patients with LN whose MPO-ANCA are positive have some difference from those with negative MPO-ANCA, but positive MPO-ANCA is not directly related to the prognosis.

Using two-kidney one-clip renal hypertensive (2K1C group), stress-induced hypertensive (neural group), DOCA-salt treated hypertensive (DOCA group) and spontaneously hypertensive rats (SHR group), to investigate the change in AT(1A)-receptor autoantibodies (AT(1A)-AAs) during the development of the four types of hypertension. The biological activities of AT(1A)-AAs were examined. It was shown that the frequency of occurrence and titres of AT(1A)-AAs increased significantly during the development of hypertension. In the four hypertensive groups studied, the occurrence of AT(1A)-AAs was most prominent in SHR, 2K1C and neural groups. The biological effects of AT(1A)-AAs were shown to increase the beating frequency of cultured neonatal myocardial and vascular contractile tension. It is suggested that autoimmune mechanisms are involved the pathogenesis of different types of hypertension and the AT(1A)-AAs may be one of the mechanisms leading to cardiac hypertrophy.
Animals ; Autoantibodies ; blood ; Desoxycorticosterone ; administration & dosage ; Hypertension ; classification ; etiology ; immunology ; physiopathology ; Hypertension, Renovascular ; immunology ; physiopathology ; Male ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Rats, Wistar ; Receptor, Angiotensin, Type 1 ; immunology ; Stress, Physiological ; physiology