A truncated mutant of the Open Reading Frame 2 (ORF2, aa384-606) was amplified from cDNA of genotype IV hepatitis E virus (HEV) by polymerase chain reaction (PCR), subcloned to expression plasmid pTO-T7, and expressed in Escherichia coli. SDS-PAGE and Western blotting were used to detect and identify the recombinant protein, namely rP24. After washing of inclusion bodies, dissolving in denaturing agents, refoldeding by dialysis, ion exchange chromatography and gel chromatography, dynamic light scatter was used to study the hydrated radius of rP24. Western blotting was applied to detect the immunoreactivity of rP24, and mouse immunity test and indirect enzyme linked immunosorbent assay (ELISA) were applied to evaluate the immunogenicity and the detection rate of HEV positive and negative serum. SDS-PAGE and Western blotting show that rP24 was highly expressed in the form of inclusion bodies after induction, and had strong immunoreactivity to monoclonal antibody (McAb) 15B2. After a multi-step purification of rP24, Western blotting indicated that the purified rP24 also had strong immunoreactivity to neutralizing McAb 8C11 and HEV positive serum, suggesting that rP24 simulated the nature structure of HEV capsid protein. Dynamic light scatter demonstrated that the average hydration radius of purified rP24 was 7.48 nm. The mouse immunity test showed that the purified rP24 also had good immunogenicity, and the period of serum antibodies converted from negative to positive was very short, but the antibodies maintained more than 20 weeks. Indirect ELISA tests showed that the detection rate of was the same as anti-HEV-IgG diagnostic kit (Wan Tai corporation). Taken together, the rP24 simulated the neutralizing epitopes of natural HEV, and had strong immunoreactivity and immunogenicity. It provided a basis for the further investigation of the difference of infection mechanism between genotype I and genotype IV HEV.
Animals ; Antibodies, Monoclonal ; Antibodies, Neutralizing ; Blotting, Western ; Capsid Proteins ; biosynthesis ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; Genotype ; Hepatitis E virus ; Mice ; Mutant Proteins ; biosynthesis ; Open Reading Frames ; Recombinant Proteins ; biosynthesis

Objective To investigate the factors implicated in the outcomes of patients with invasive pulmonary aspergillosis (IPA).Methods During a 5-year period,65 patients with IPA met the criteria set by the European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG)in 2008 were retrospectively evaluated. The initial CT findings of eligible patients were reviewed by two senior radiologists who specialized in chest radiology.Patients were divided into the survivor (n =43 ) and non-survivor (n =22) groups according to their survival as long as 3 months after the diagnosis of IPA was made.An initial univariate analysis was used to screen variables that were related to prognosis,followed by a multivariate logistic regression analysis to examine these variables. Results Of the 65 IPA patients analyzed,23 (35％) had a proven diagnosis and 42 (65％) were probable ones.The univariate analysis showed that the rates of extra-pulmonary infection,uncontrolled underlying diseases and invasive mechanical ventilation were significantly different between the 3-month survival group and the non-survival group ( P ＜0.05,respectively),whereas chest CT findings,including air-space consolidation/massive consolidation,macronodules,infarct-like macronodules,halo signs, ground-glass opacities,small nodules,hypodense signs,cavities,crescent signs,small-airway findings,bronchial wall thickening/bronchiectasis,pleural effusion and hydro-pericardium, were not significantly different between the two groups (P ＞ O.05,respectively).Logistic regression analysis revealed that an uncontrolled underlying disease was the only independent predictor of 3-month mortality in patients with IPA (P =0.001,OR:O.024,95 ％ CI:O.003 ～0.223,B =- 3.714,SE =1.129,Wald =10.821 ). Conclusions An uncontrolled co-morbidity was the only independent predictor of mortality within 3 months in patients with IPA.The initial CT findings did not confer any informatioin of implication in predicting the outcomes of IPA patients.

Objective There is little research focusing on the expression and function of bradykinin 1 receptor ( B1R ) and bradykinin 2 receptor ( B2R) after cerebral ischemia/reperfusion on the basis of diabetes .The aim of this study was to compare the ex-pression difference and function change of B 1R and B2R in non-dia-betic and diabetic rats . Methods The cerebral ischemia/reperfu-sion model was established on 41 non-diabetic and type 2 diabetic rats, the weight and the biochemical index were measured on these two types of rats .8 non-diabetic rats and 8 diabetic rats were respec-
tively assigned to two groups according to random number tables:control group and I/R 24 h group, 4 in each group.Real-time PCR was performed to observe the expressions of two receptors at 24 h after reperfusion .Then, 33 non-diabetic rats and 33 diabetic rats were randomly divided into 4 groups respectively, including sham group (n=6), saline group (n=9), B1R antagonist group (n=9) and B2R antagonist group (n=9).At 24 hours after cerebral I/R, neurological deficiency was evaluated by neurological severity scores ( NSS);infarct volume was observed by TTC staining;cell apoptosis was determined by TUNEL staining;neuron degeneration was de-tected by Fluoro-Jade C staining. Results Glucoses of diabetics at 3, 7, 14 d after model establishment [(23.45 ±5.01), (23.71 ±4.87), (22.72 ±4.11) mmol/L] were obviously elevated compared with non-diabetics [(5.77 ±0.75), (6.05 ±0.69), (7.15 ±1.09) mmol/L];blood cholesterin [(4.59 ±3.43) mmol/L] and insulin [(67.26 ±12.02) pmol/L] at 14 d after model establishment were evidently incresaed in comparison to those in non-diabetics [(1.58 ±0.37) mmol/L, (25.34 ±4.88) pmol/L] (P<0.05), while no significant difference was found in the blood triglyceride of diabetics between them (P>0.05).Compared with non-diabetics, diabetics suffered from more apparent up-regulation of B1R mRNA (P<0.01) but relatively less B2R mRNA (P<0.05) at 24 h after I/R.NSS score, infarction volume, damaged and apoptotic cells in B2R antagonis-treated non-diabetic rats at 24 h after I/R conspicuously decreased compared with saline-treated non-daibetic rats.Those indicators in B1R antagonis-treated diabeics were strikingly lessened compared with saline-treated daibetics . Conclusion I/R induced distinct up-regulation of B2R mRNA in non-diabetics and inhibiton of B 2R effectively ameliorated the infarct volume and cell injury after I/R in non-diabetics; I/R induced more notable up-regulation of B1R mRNA in diabetics and B1R antagonist exerted neuroprotective effects instead of B 2R antagonist af-ter I/R in diabetics.

Objective To investigate the effect of thyroxine replacement therapy with residual subclinical hypothyroidism on the success rate of catheter ablation in elderly patients with atrial fibrillation(AF).Methods Among the consecutive patients with AF who underwent a first AF ablation in our center between 2009 and 2012,we identified 56 patients(41 paroxysmal AF,15 persistent AF)with subclinical clinical hypothyroidism after receiving thyroid hormone replacement therapy as study group.The control group consisted of 56 patients with euthyroidism and no history of thyroid dysfunction.All patients underwent catheter ablation.Results At the end of follow up,37.5%(21/56)patients were AF free after the first procedure in the study group,in comparison to 64.3%(36/56)in control group(χ2=8.655,P=0.003).Last procedure was performed in 27 patients of study group and in 15 patients of control group.After the last performed ablation,62.5%(35/56)study group patients and 80.4%(45/56)controls group patients had no recurrence(χ2=4.653,P=0.031).The major complications rate did not differ between two groups(P=0.642).Conclusions Thyroid hormone replacement therapy with residual subclinical hypothyroidism reduces catheter ablation success rate in elderly patients with atrial fibrillation.

PURPOSE: The goal of this study was to purify and characterize Ebola virus glycoprotein (GP)-specific IgG antibodies from hybridoma clones. MATERIALS AND METHODS: For hybridoma production, mice were injected by intramuscular-electroporation with GP DNA vaccines, and boosted with GP vaccines. The spleen cells were used for producing GP-specific hybridoma. Enzyme-linked immunosorbent assay, Western blot assay, flow cytometry, and virus-neutralizing assay were used to test the ability of monoclonal IgG antibodies to recognize GP and neutralize Ebola virus. RESULTS: Twelve hybridomas, the cell supernatants of which displayed GP-binding activity by enzyme-linked immunosorbent assay and the presence of both IgG heavy and light chains by Western blot assay, were chosen as a possible IgG producer. Among these, five clones (C36-1, D11-3, D12-1, D34-2, and E140-2) were identified to secrete monoclonal IgG antibodies. When the monoclonal IgG antibodies from the 5 clones were tested for their antigen specificity, they recognized GP in an antigen-specific and IgG dose-dependent manner. They remained reactive to GP at the lowest tested concentrations (1.953–7.8 ng/mL). In particular, IgG antibodies from clones D11-3, D12-1, and E140-2 recognized the native forms of GP expressed on the cell surface. These antibodies were identified as IgG1, IgG2a, or IgG2b kappa types and appeared to recognize the native forms of GP, but not the denatured forms of GP, as determined by Western blot assay. Despite their GP-binding activity, none of the IgG antibodies neutralized Ebola virus infection in vitro, suggesting that these antibodies are unable to neutralize Ebola virus infection. CONCLUSION: This study shows that the purified IgG antibodies from 5 clones (C36-1, D11-3, D12-1, D34-2, and E140-2) possess GP-binding activity but not Ebola virus-neutralizing activity.
Animals ; Antibodies* ; Antibody Formation ; Blotting, Western ; Clone Cells ; Ebolavirus* ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Glycoproteins* ; Hemorrhagic Fever, Ebola ; Hybridomas ; Immunoglobulin G* ; In Vitro Techniques ; Mice ; Sensitivity and Specificity ; Spleen ; Vaccines ; Vaccines, DNA

OBJECTIVETo study the method and effect of endoscopic two-portal one-way releasing procedure for cut of transverse carpal ligament and decompression of median nerve.

METHODSEleven female patients (13 sides) with primary carpal tunnel synrome underwent endoscopic two-portal one-way releasing procedure, there were 3 left hands, 6 right, and 2 both. All the subjects had hypesthesia in the radial three and half finger's tip with a positive, Tinel sign of median nerve at wrist; 11 cases had thenar myatrophy in which 4 had opposition dysfunction. Under local anaesthesia, the proximal incision was located at the point of the proximal carpal transverse striation level between palmaris longus and flexor carpi radialis. The outlet was chosed the junction of the parallel line of the ulnar side of thumb and proximal extending line of middle ring fingers' long axis while the thumb was in abduction position. The length of each incision was only one centimeter. The hook knife was inserted to the proximate of the transverse carpal ligament, then, the transverse carpal ligament was completely released form the proximal to the distal end by hook knife under the endoscope monitor.

RESULTSThe results showed that both pinch and grip function was satisfied and no complications occurred at 4 to 20 months followed-up. S3+ M3 or more has been reached in 3 months after operation.

CONCLUSIONThe endoscopic two-portal one-way releasing procedure is an easy and effective method for the treatment of carpal tunnel syndrome.

Adult ; Aged ; Carpal Tunnel Syndrome ; pathology ; physiopathology ; surgery ; Decompression, Surgical ; Endoscopy ; methods ; Female ; Follow-Up Studies ; Humans ; Ligaments ; surgery ; Median Nerve ; surgery ; Middle Aged ; Recovery of Function

Objective To evaluate the clinical effect of Jianpi tiaozhong powder in the treatment of children with functional dyspepsia ( FD) and its effects on the gastric motility.Methods Ninety children with FD were randomly divided into treatment group and control group , 45 cases in each group.Children in treatment group were given Jianpi tiaozhong powder one hour after the breakfast and supper , 10 mL for five to nine years old children and 20 mL for ten to fourteen years old children.Meanwhile , children in control group were given domperidone syrup orally, the doses of 5-7, 8-10 and 11-14 years old children were 5, 7 and 10 mL, separately.After one -month treatment, the clinical symptoms, side effect, electrogastrogram(EGG), motilin(MOT), soma-tostatin(SS) and gastrin(GAS) were recorded and compared before and after treatment.Results After treatment , the total efficiency of treatment group was 86.67%, significantly higher than that in control group, which was 75.56%( P<0.05 ).Improvement of symptoms in treatment group was more significant than that in control group( P<0.05 ).The percents of normal rhythm before meal were significantly higher than before treatment ( P<0.05 ).The motilin and somatostatin in treatment group were ( 235.7 ±19.4 ) , ( 18.8 ±2.5 ) pg · mL-1 , significantly lower than those in control group , which were ( 267.3 ±17.2 ) , ( 20.3 ±1.4 ) pg · mL-1.Gastrin in treatment group and control group were (168.6 ±17.2), (142.8 ±11.4) pg· mL-1, respectively (P<0.05).The motilin, somatostatin and gastrin after treatment were comparable than those before treatment in two groups ( P <0.05 ).There was no adverse drug reaction in two groups.Conclusion Jianpi tiaozhong powder had precise efficacy and slight side effect on children with FD.

OBJECTIVETo evaluate the effect of Helicobacter pylor (Hp) eradication in children with acute idiopathic thrombocytopenic purpura (ITP).

METHODSNinety-three children with acute ITP and Hp infection were divided into two groups and treated with prednisone and Hp eradication (group A, 51 cases) or with prednisone without Hp eradication (group B, 42 cases).

RESULTSThe Hp eradication rate was 94.1% in group A. No difference was found in the therapeutic effects on IPT between the two groups, but the recurrence rate in one year in group A was significantly lower than that in group B.

CONCLUSIONNHp eradication does not obviously enhance the therapeutic effect on childhood acute ITP, but can decrease the relapse rate in one year. HP eradication therapy is recommended in children with acute ITP and Hp infection.

Adolescent ; Anti-Bacterial Agents ; therapeutic use ; Child ; Child, Preschool ; Drug Therapy, Combination ; Female ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; drug effects ; immunology ; Humans ; Male ; Prednisone ; therapeutic use ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; microbiology

Objective:To evaluate the risk factors for adjacent vertebral compression fracture (AVCF) after fusion surgery of lumbar canal stenosis in the elderly.Methods:A retrospective cohort study was designed for 297 patients with lumbar canal stenosis who underwent fusion surgery in Hubei Provincial Hospital of Integrated Chinese and Western Medicine from January 2017 to December 2020. There were 42 males and 255 females, aged 68-85 years [(76.3±7.2)years]. The patients were divided into AVCF group ( n=67) and non-AVCF group ( n=230) according to with or without the occurrence of AVCF. The indicators recorded were the gender, age, body mass index, education level, number of combined basic diseases, course of the disease, preoperative bone mineral density, intraoperative bleeding volume, number of level fused during operation, postoperative blood transfusion volume, postoperative observation in the ICU, types of postoperative external fixation and time of external fixation. Univariate analysis was conducted to analyze the correlation between above risk factors and AVCF after fusion surgery of lumbar canal stenosis in the elderly. Multivariate Logistic regression analysis was used to determine the independent risk factors for the occurrence of AVCF in these patients. Results:In the univariate analysis, age, education level, preoperative bone mineral density, intraoperative bleeding volume, postoperative blood transfusion volume and postoperative observation in the ICU were correlated with AVCF after fusion surgery of lumbar canal stenosis in the elderly ( P<0.05 or 0.01), while there was no correlation of AVCF with gender, body mass index, number of combined basic diseases, course of the disease, number of level fused during operation, types of postoperative external fixation and time of external fixation (all P>0.05). Multivariate Logistic regression analysis showed that gender ( OR=4.02, 95% CI 1.35, 12.00, P<0.05), preoperative bone mineral density≤-2.5 SD ( OR=2.01, 95% CI 1.47, 2.75, P<0.01), intraoperative bleeding volume≥475 ml ( OR=1.01, 95% CI 1.00, 1.01, P<0.01) and postoperative blood transfusion volume≥434 ml ( OR=0.99, 95% CI 0.98, 1.00, P<0.01) were significantly associated with AVCF after fusion surgery of lumbar canal stenosis in the elderly. Conclusion:The female, bone mineral density≤-2.5 SD, intraoperative bleeding volume≥476 ml and postoperative blood transfusion volume≥434 ml are independent risk factors for AVCF after fusion surgery of lumbar canal stenosis in the elderly.

OBJECTIVETo investigate whether the MTNR1A gene promotor polymorphism (rs2119882) are associated with the occurrence or curve severity of adolescent idiopathic scoliosis (AIS).

METHODS226 AIS patients and 277 normal controls were recruited. The maximum Cobb angles were recorded in AIS patients. PCR-RFLP was used for the genotyping.

RESULTSThe genotype and allele frequency distribution were comparable between AIS and normal control, the mean maximum Cobb angle of different genotypes of rs2119882 were similar with each other among AIS patients.

CONCLUSIONThe MTNR1A gene promoter polymorphism was neither associated with the occurrence nor the curve severity of AIS.

Adolescent ; Adult ; Child ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Receptor, Melatonin, MT1 ; genetics ; Scoliosis ; genetics