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 immunostaining of endothelin-1(ET-1) and von Willebrand factor(vWF) in pulmonary vascular of ventricular septal defect(VSD) with pulmonary hypertension(PH),and the possible roles of ET-1 and vWF in the development of PH and remodeling in pulmonary artery.Methods VSD with non-PH were choosed as control group,20 cases with VSD with PH were divided into Ⅰgroup and Ⅱgroup according to mean pulmonary artery pressure. The specimens from middle lobe of right lung was obtained and stained by immunohistochemical technique.The degree of immunoreaction was measured with light density.Results The light density of ET-1 and vWF in small pulmonary arteries had significant statistical difference between every group,and Ⅱgroup was higher than those of Ⅰgroup,but no difference in small pulmonary veins.In every group,the light density of ET-1 and vWF had significant difference between small pulmonary arteries and pulmonary veins.There were statistical correlation among mean pulmonary pressure,VSD/A,ET-1 and vWF.Conclusions ET-1 and vWF are concerned with the development of PH and mediate pulmonary vascular remodeling.The data of small pulmonary veins suggest that the main origin of ET-1 in pulmonary tissue is small pulmonary artery.

On the basis of the development of a new CE method for the analysis of α-lactalbumin (α-Lac), β-lactoglobulin A(β-LgA) and β-lactoglobulin B(β-LgB), different concentrations of acetic acid (HAc) and trichloroacetic acid (TCA) for the removal of proteins in raw milk, infant formula A and B, and yogurt were investigated.It was shown that TCA is better than HAc for eliminating of proteins in the above four samples.The proteins in 2 g of raw milk could be eliminated by 3 mL of 100 g/L TCA.0.5 g of infant formula A and B needs 5 mL of 10 g/L and 20 g/L TCA to remove the proteins, respectively.For 2 g of yogurt, 3 mL of 20 g/L TCA is enough.The present research may provide valuable information for protein-containing sample (such as raw milk, infant formula and yogurt) pretreatment.

OBJECTIVE: Mitochondrial deoxyribonucleic acid (mtDNA) 8344 A>G (m.8344A>G) mutation is the common mutation associated with mitochondrial myoclonus epilepsy with ragged-red fibers (MERRF) syndrome. Herein we report a rare case with mitochondrial encephalopathy, lactic acidosis and stroke-like episodes/MERRF/Leigh (MELAS/MERRF/Leigh) overlap syndrome caused by m.8344A>G mutation. METHODS: The clinical and imaging data of the patient were collected and an open muscle biopsy was carried out. We further employed molecular genetic analyses to detect mtDNA mutation in the proband and his mother. And then a clinical and neuroimaging follow-up was performed. RESULTS: This patient was a 25-year-old male, who developed exercise intolerance since the age of 6. At age 10, he suffered from acute episodes of hemianopia, and cranial magnetic resonance imaging (MRI) showed occipital stroke-like lesions and cranial magnetic resonance spectroscopy (MRS) revealed a lactate peak corresponding to the lesion. After that the patient presented slowly progressive psychomotor decline. He had myoclonic seizures and cerebellar ataxia since the age of 12. At age 21, he was admitted to our hospital because of confusion and cranial MRI revealed symmetrical lesions in bilateral posterior putamen, thalami and midbrain. Then repeated MRI showed progression of original lesions and new frontal multiple stroke-like lesions. Symptomatic and rehabilitation treatment relieved his condition. Follow-up cranial MRI at age 24 showed the lesions in basal ganglia and thalami diminished, and the midbrain lesions even completely vanished. Muscle pathology indicated the presence of numerous scattered ragged-red fibers (RRF), suggestive of a mitochondrial disorder. Polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP) detected the m.8344A>G mutation of the MT-TK gene encoding mitochondrial transfer RNA for lysine in the patient's blood. Next generation sequencing (NGS) of the whole mitochondrial genome identified that the proportion of m.8344A>G was 90%, and no other mtDNA mutation was detected. Sanger sequencing further identified this mutation both in the proband and his mother's blood, although the mutation load was much lower in his mother's blood with approximately 10% heteroplasmy. CONCLUSION The present study is the first to describe a patient with m.8344A>G mutation in association with the MELAS/MERRF/Leigh overlap syndrome, which expands the phenotypic spectrum of the m.8344A>G mutation.
Acidosis, Lactic ; Adult ; Child ; DNA, Mitochondrial/genetics* ; Humans ; Male ; Mitochondrial Encephalomyopathies ; Mutation ; Stroke ; Young Adult

Objective Multimeric cyclic RGD (Arg-Gly-Asp) peptides are capable of improving the integrin αvβ3-binding affinity due to the polyvalence effect.In this study,the authors prepare 99Tcm-la-bearing cyclic RGD tetramer E{E[c(RGDfK)]2}2,and evaluate its biodistribution and imaging in nude mice beating U87 MG human glioma xenografts with integrinαvβ3-positive.Methods 99Tcm-hydrazino-nictinamide (HYNIC)-E{E[c(RGDfK)]2}2 was prepared by two-step method,while HYNIC wag chosen as bifunctional chelator,and tricine and trisodium triphenylphosphine-3,3,3-trisuifonate (TPPTS) as coligands.The af-finity of c (RGDyK) monomer,HYNIC-E[c(RGDfK)]2 dimer and HYNIC-E{E[c(RGDfK)]2}2 tetramer to integrin αvβ3 was compared by in vitro competitive assay against binding of 125I-c(RGDyK)to integrin αvβ3.positive U87 MG human glioma cells.The biodistribution [the percentage of injection dose per gram of tissue(%ID/g)] and imaging were performed in nude mice bearing UB7MG human glioma xenografts.Re-suits The labeling yield of 99Tcm-HYNIC-E{E[c(RGDfK)2}2 was over 95%,and the radiochemical purity was more than 99%after purification with Sop-Pak C18 cartridge.The 50%inhibiting concentration (IC30) val-ues of c(RGDyk),HYNIC-E[c(RGDfK)]2 and HYNIC-E{E[c(RGDfK)]2}2 were 85.9,9.5 and 4.5 nmol/L, respectively.The result indicated that RGD tetramer possessed a significantly higher affinity to in-tegrinαvβ3.The biodistribution data showed that 99Tcm-HYNIC-E{E[c(RGDfK)]2}2 was excreted mainly through kidneys.The tumor uptake of 99Tcm-HYNIC-E{E[c(RGDfK)]2}2 was two times higher than 99Tcm- HYNIC-E[c(RGDfK)]2,at 1h postinjection,with the uptake of(10.32±0.07)%ID/g and(5.15±0.52)%ID/g,respectively,which was consistent with the in vitro competitive binding data.The tumor up-tale of 99Tcm-HYNIC.E{E[c(RGDfK)]2}2 was still as higher as(9.35±1.35)%ID/g at 4 h postinjec-tion, which demonstrated that the retention time of radiotracer in tumor was long enough.The imaging showed that tumor was clearly visualized at 1h postinjection,and the image at 4 h postinjection Was better.Conclusion The higher tumor uptake and longer retention time in tumor make 99Tem-HYNIC-E{E[c(RG-DfK)J 2}2 a promising radiotracer for integrinαvβ3-positive tumors imaging,furthermore,suggest that radi-onuelides(such as 90Y).1abeled RGD tetramer is more suitable for the therapy of integrin αvβ3-positive tumors.

The chemical investigation on Ganoderma philippii led to the isolation of sixteen compounds by silica gel and Sephadex LH-20 column chromatography. On the basis of spectroscopic data analyses, their structures were elucidated as 2, 5-dihydroxyacetophenone (1), methyl gentisate (2), (S) -dimethyl malate (3), muurola-4, 10 (14) -dien-11beta-ol (4), dihydroepicubenol (5), 5-hydroxymethylfuran carboxaldehyde (6), ergosta-7, 22E-dien-3beta-ol (7), ergosta-7, 22E-dien-3-one (8), ergosta-7, 22E-diene-2beta, 3alpha, 9alpha-triol (9), 6/beta-methoxyergo-sta-7, 22E-dien-3beta, 5alpha-diol (10), ergosta-4, 6, 8(14), 22E-tetraen-3-one (11), ergosta4, 6, 8-(14), 22E-etetraen-3beta-ol (12), 5alpha, 8alpha-epidioxy-ergosta-6, 22E-dien-3beta-ol (13), 7alpha-methoxy-5alpha, 6alpha-epoxyergosta-8-(14), 22E-dien-3beta-ol (14), ergosta-8, 22E-diene-3beta, 5alpha, 6beta, 7alpha-tetraol (15), and ergosta-5, 23-dien-3beta-ol, acetate (16). All the compounds were obtained from this fungus for the first time, and compounds 4 and 5 were isolated from the Ganoderma genus for the first time.
Ganoderma ; chemistry ; Medicine, Chinese Traditional ; Organic Chemicals ; analysis ; isolation & purification

Objective To evaluate the methods and consequences of surgical technique in the treatment of Stanford A aortic dissection.Methods 108 patients with type Standford A aortic dissection underwent surgery in our study,including urgent surgery in 53 and selective surgery in 55.The operation was performed under deep hypothennic circulatory arrest (DHCA) in 85 cases.Surgical procedures included ascending and semi arch replacement or total arch replacement (some cases combined with stented graft implanted into the descending aorta),"elephant trunk" procedure.Concomitant procedures included repair of intimal tear in arch or descending aorta,Bentall procedure,aortic valve replacement,Cabrol or modified Cabrol procedure,aortic valvuloplasty,mitral valvuloplasty or mitral valve replacement,tricuspid valvuloplasty and CABG.Results In-hospital mortality was 6.5% (7 of 108 patients).The mor- tality was 7.5% (4 of 53 patients) in urgent surgery group and in elective surgery group was 5.4% (3 of 55 patients).Ninety six percent survived patients were followed up for 1 month to 13.3 years [mean (3.2?1.3) years] and 2 deaths occurred during the fel- low-up period.3 patients underwent re-operatian.Conclusion The choice of surgical procedures depend on the location of intimal tear for Stanford A aortic dissection.The better operative effects can be expected with proper surgical indication,perfecting surgical technique,and enhancing postoperative treatment.

Objective To study the antigenicity of human thyrotropin receptor(hTSHR)amino terminus (amino acid 29～280)and its association with Graves' disease.Methods Total thyroid RNA was prepared from human normal thyroid tissue.RNA was then reversely transcripted and cDNA was subjected to PCR amplification.PCR product was cloned into pcDNA3.1 and the recombinant plasmid was named pcDNA3.1/hTSHR188～940bp. Balb/c mice were immunized with peDNA3.1/hTSHR188～940bp. The levels of serum thyroxin,anti-TSHR antibody(TRAb)and thyroid stimulating antibody(TSAb)were measured,and the pathological changes of thyroid tissue were also observed.Results A 753 bp fragment encoding hTSHR ectodomain amino end was obtained after PCR amplification.Confirmed by Hind Ⅲ restriction enzyme digestion and DNA sequencing,pcDNA3.1/hTSHR188～940bphad been constructed successfully,with the correct sequence and direction of hTSHR188～940bp.In the Balb/c mice treated with pcDNA3.1/hTSHR188～940bp,elevated TRAb in week 6(0.148±0.018)were observed compared with those at week o(0.106±0.006,P＜0.01),and kept a higher level till week 10(0.134±0.011,P＜0.01).T4 and TSAb index values were significantly increased in week 10.Serum T4 concentration increased from(41.02±7.97)μg/L in week 0 to(62.20±12.77)μg/L in week 10(P＜0.01);TSAb index values rose from 0.864±0.076 at week 0 to 1.392±0.615(P＜0.01).Thyroid pathological examination showed that proliferated thyroid follicular epithelial cells and foll icular eapacity increased.Inflammatory cells were occasionally found.Conclusions There are antigen epitopes in hTSHR ectodomain amino acid 29～280,which can stimulate the production of TSAb.And the latter induces hyperthyroidism and Graves' disease like manifestations.It suggests that hTSHR ectodomain amino acid 29～280 is closely associated with Graves' disease,and maybe one of important etiological factors leading to the disease.

OBJECTIVETo apply needle-knife to treat lumber disc herniation (LDH) and surface electromyography were used to analyze biomechanical characteristic of patient's lumber muscle to make a comprehensive evaluation on its efficacy.

METHODSThirty patients who met the inclusive criteria were selected and treated with needle-knife, once a week for 2 weeks. Visual analogue scale (VAS), ASLR and JOA score before and after treatment were observed. Surface electromyography was applied to test the surface electromyography signals. AEMG, MFs and MPF were calculated before and after the treatment.

RESULTSAfter treatment, VAS was significantly reduced, ASLR, JOA, AEMG and MPF were obviously increased, and the absolute value of MFs was lowed (all P < 0.01).

CONCLUSIONThe needle-knife could significantly relieve lumbar muscle strength, muscle tone and muscle fatigue, improve in the imbalance of lumbar extensor muscle group, leading to the recovery of biomechanical characteristic, and the clinical efficacy is superior.

Acupuncture Therapy ; Adult ; Electromyography ; Female ; Humans ; Intervertebral Disc Displacement ; diagnosis ; physiopathology ; therapy ; Lumbar Vertebrae ; physiopathology ; Male ; Middle Aged

It was difficult to prepare traditional Chinese medicine pellets due to the adverse characteristics of the herbal extract. In this study, Danshen extract (DS) powder mixed with different proportions of microcrystalline cellulose (MCC), lactose and starch were made into pellets by extrusion-spheronization. Particle size, span, bulk density, tapping density, compressibility, Hausner ratio and angle of repose were used to evaluate the micromeritic properties of mixing powders. Feret diameter, aspect ratio, yield, density and friability were used to evaluate the properties of the pellets. The correlations between micromeritic properties of raw material powders and the formability of their pellets were analyzed by cluster analysis, principal component analysis and partial least squares regression analysis. As a result, the particle size of the powders was negatively correlated with the size, density, yield, and was positively correlated with the friability of their pellets. The span, density, compressibility and angle of repose of the powders were positively correlated with the size, density, yield, and were negatively correlated with the friability of their pellets. So there were certain correlations between the micromeritic properties of raw material powders and the properties of their pellets prepared by extrusion-spheronization. This research provided a foundation for the technology and method of traditional Chinese medicine extract pellets.
Cellulose ; chemistry ; Drug Compounding ; methods ; Drug Implants ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Excipients ; chemistry ; Lactose ; chemistry ; Medicine, Chinese Traditional ; methods ; Particle Size ; Powders ; chemistry ; Starch ; chemistry