Objective To set up an antibody-capture ELISA method to detect the Japanese encephalitis virus(JEV)antibody.Methods ELISA plate was coated with the monoclonal antibody which was specific to the envelope protein epitope E39 of JEV,JEV SA14-14-2 strain as the source of antigen was used to absorb the monoclonal antibody,the absorbed virus used to capture the JEV'S antibody.The antibody that captured ELISA was established.The indirect ELISA method using the virus particles from cell culture was compared with coating ELISA plate,105 clinical serum were checked.Results The background in indirect ELISA assay could not be abscised,positive and negative serum diluted in a ratio of 1:10,1:100,1:1000,the relative value of A posative/A negative were 1.02,0.99,1.13,all<2.1.But the antibody-captured ELISA method when the serum dilution was 1:10,1:100,the A posative/A negative were 3.57,2.94,all>2.1;when the dilution was 1:1000,the A posative/A negative was 1.42,<2.1,it meant the method could distinguish the positive and negative serum efficiently when the dilution Was 1:100,the background problem in indirect ELISA assay could be solved.Antibody-capture method was used to check 105 serum samples,the A posative/A negative over a range of 0.257～0.321(0.262±0.050),all<2.1,no positive sample found.Conclusion The antibody-capture ELISA method has been preliminary set up with a high specificity,capable of quickly identifying JEV from other virus.

BACKGROUND: BuCy2 composed of large-dose oral busulfanum and cyclophosphamide has been one of the main conditioning regimens before transplantation, but oral busulfanum can result in many side effects.OBJECTIVE: To evaluate the efficacy and safety of the conditioning regimen with intravenous busulfanum (Ⅳ Bu) in hematopoietic stem cell transplantation (HSCT) for malignant hematopathy.DESIGN: Case observation.SETTING: Department of Hematology, Union Hospital,Tongji Medical College, Huazhong University of Science and Technology.PARTICIPANTS: A total of 13 patients with chronic myelogenous leukemia (CML) and 6 patients with acute leukemia,who received allogeneic HSCT with the conditioning therapy of modified BuCy2, were enrolled at Institute of Hematopathy, Union Hospital, Huazhong University of Science and Technology from May to December 2006. There were 12 males and 7 females aged 14-50 years with an average of 33 years, and there were 9 cases of related transplantation and 10 cases of unrelated transplantation. Hematopoietic stem cells (HSCs) were harvested from peripheral blood of 18 subjects and bone marrow of 1 subject. All patients and their family members signed the informed consent.METHODS: All patients were treated with allogeneic HSCT with the conditioning therapy of intravenous BuCy2.All patients received the modified combination of busulfanum and cyclophosphamide that was commonly utilized currently.Hydroxycarbamide 40 mg/kg was orally taken ten days before transplantation. Arabinosylcytosin (Ara-C) 2 g/m2 was injected via vein nine days before transplantation. Busulfanum ampule 0.8 mg/kg (equal to 1 mg/kg oral administration)was given by central venous cannula eight, seven and six days before transplantation. The injection was performed over 2 hours controlled by infusion pump, once every 6 hours, totally 12 times. Busulfanum was diluted by saline or 5% glucose to about 0.5 g/L (about 10 times) before Ⅳ Bu. Cyclophosphamide 1.8 g/m2 was intravenously injected every day from five and four days before transplantation. Methyl-CCNU 250 mg/m2 was taken orally three days before transplantation. Occurrence of hepatic vano-occlusive disease (HVOD) and its side effects were determined. Follow-up was performed at month 6.5 after operation to observe disease recurrence and patient survival.MAIN OUTCOME MEASURES: Occurrence and side effects of HVOD 100 days after transplantation, and results of follow-up.RESULTS: Totally 19 patients were involved in the result analysis. In 13 days after transplantation, blood conversion,chromosome karyotype and DNA polymorphism tests verified that reconstruction of hematopoietic function was obtained and two patients got recurrence, of which one patient got complete remission after receiving fludarabine plus cytarabine plus granulocyte colony-stimulating factor (G-CSF) and HSCs from peripheral blood. DNA polymorhism test verified that re-engraftment was obtained. Another patient received chemotherapy. Other patients lived well.

0.05).The methyla tion rates were higher in tumor tissue than those in normal and adjacent mueosa(P

Objective To evaluate the application value with low-dose CT scan and quantitative analysis in airway disease of chronic obstructive pulmonary disease(COPD).Methods All subjects(34 control cases,88 cases of stable COPD patients)underwent low-dose CT chest scans,and 88 COPD patients underwent pulmonary function tests.Measuring the airway dimensions of 3th generation in apical bronchus of the right upper lobe,the posterior basal bronchus of the right lower lobe and left lower lobe.Results COPD patients with moderate to extremely severe airflow limitation demonstrated higher WA% and Pi10 in comparison with normal volunteers(all P<0.05),meanwhile patients with extremely severe airflow limitation demonstrated higher WA % and Pi10 in comparison with patients with mild airflow limitation(both P <0.05).COPD patients with severe airflow limitation demonstrated higher T/BSA and WA/BSA in comparison with normal volunteers and patients with moderate airflow limitation (all P<0.05).The WA % and Pi10 of COPD patients group were negatively correlated with ventilation indices and small airway indices(all P <0.01).Pi10 were positively correlated with lung volume indices (all P < 0.05 ).Conclusion Low-dose CT scan and quantitative analysis can provide an objective and quantitative method to evaluate the information about airway disease of COPD,inwich both WA % and Pi10 are objective indices.With the increase of air flow limitation,the bronchial wall thickness gradually increased.

OBJECTIVE To investigate the productive rate of AmpC ?-lactamases and ESBLs produced by the Gram-negative bacilli from the nosocomial infection in our hospital. METHODS AmpC ?-lactamases and ESBLs were detected by the improved cefotaxime and ceftriaxone three-dimensional test. RESULTS The productive rate of AmpC ?-lactamases was 16.00%.Among them,the productive rate of AmpC ?-lactamases was only 8.84%.The Enterobacter cloacae,Serratia marcescens and Enterobacter aerogenes were easy to produce the enzymes(36.00%,31.25% and 28.00%).The productive rate of AmpC ?-lactamases and ESBLs at the same time was 7.16%. CONCLUSIONS The productive rate of the enzymes by the Gram-negative bacilli from the nosocomial infection is rather high.

Abstract: Objective To investigate the dynamic changes and clinical significance of specific antibodies in patients with coronavirus disease 2019 (COVID-19). Methods A retrospective study was conducted to collect 141 adult COVID-19 survivors who were followed up in the Eighth Hospital affiliated to Guangzhou Medical University from February 6, 2020, to March 24, 2021. The patients were divided into severe group (severe and critical) and non-severe group (light and ordinary) according to the diagnosis at discharge. The antibody changes of the two groups were compared and analyzed at 1 week, 2 weeks, 1 month, 3 months, 6 months and 1 year after discharge. Results After discharge from hospital, the positive rate of IgG in the severe group was 95.00% after 1 week and 100.00% in the following year, in the positive rate of IgG in the non-severe group was 59.50% after 1 week, 90.08% in 6 months and 76.03% in one year. The level of serum IgG in the severe group was significantly higher than that in non-severe group (Z=-2.441, P=0.015). One-year follow-up: the serum IgG in the severe group was significantly higher than that in the non-severe group (Z=-3.410, P=0.001). The serum IgM level of the severe group after one year follow-up was lower than that of the six months follow-up, the difference was statistically significant (Z=-2.259, P=0.024). The serum IgG and IgM level of the non-severe group after one year follow-up was lower than that of the six months follow-up, the difference was statistically significant (Z=-7.37, P<0.01; Z=3.850, P<0.01). Conclusion The level of serum protective antibody in COVID-19 patients remained high within 6 months after discharge, and remained stable within 1 year after discharge. The antibody titers in the severe group were significantly higher than those in the non-severe group and lasted for at least one year. COVID-19 survivors receive 1 year of natural immune protection, and patients with critical conditions receive immunity for longer periods of time.

The mainly antigenic sites for the adenovirus neutraliation are present on Loop1 and Loop2 of hexon.Majority research were focus in the human adenovirus.Little was known on infectious canine hepatitis virus (ICHV), which was also called canine adenovirus typeⅠ.Here,ICHV (the isolated strain) DNA was isolated and purified from the cultured MDCK cells.The Loop1 and Loop2 fragments were amplified by polymerase chain reaction(PCR) method,and then was connected by ligase T4.The target fragment was then connected with vector pET28a.The nucleotide sequence ecoding Loop1 and Loop2 was determined.The nucleotide sequence identity of Loop1 region between the isolated strain and CLL, RI261 and Toronto A26/61 strains is 100%, 100% and 83.8%, and the nucleotide sequence identity of Loop2 region between the isolated strain and CLL, RI261 and Toronto A26/61 strains is 88.1% , 88.1% and 99.3%, and amino acid identity is 93.6%, 93.6% and 98.6%.The recombinant Loop protein was expressed in E.coli and was approximately 36kDa in size,and then was purified. Then BALB/c mice were injected subcutaneously in the back and armpit with the recombinant Loop protein.The anti-ICHV antibody titers of immunized serum was tested by indirect ELISA and the titers were up to 1:320.Western blot demonstrated that immunized sera could specifically combine with ICHV. The research laid a foundation for creating new genetic engineering products of infectious canine hepatitis virus.

Objective To investigate the apearance and characteristics of skin rashes in patients suffering from dengue fever.Methods Clinical data were retrospectively analyzed for 126 hospitalized patients with dengue fever collected from August to October,2007.Results Out of 126 patients with dengue fever,69.0% (87/126)had skin rashes,which usually developed from day 1 and 11 after fever.About 59.7%(52/87)of these patients developed skin rashes from day 3 to 7 after fever.No eruption order was evidenced in 59(67.8%)patients.Skin rashes were located in both the trunk and limbs in 38(43.7%)patients,and only in the limbs in 30 (34.5%)patients.The incidence rates of maculae,maculopapules,papules.hemorrhagic rash and mixed eruptions were 14.9%(13/87),14.9%(13/87),18.4%(16/87),26.4%(23/87)and 21.8%(19/87),respectively.Conclusions Most patients with dengue fever developed skin rashes from day 3 to 7 after fever.Limbs and trunk are predilection sites of skin rashes in dengue fever.Skin rashes in dengue fever is complex,including maculae,papules,hemorrhagic rashes,vesicles and mixed rashes.

With comparison of three different methods for the marking of amines compound, an optimal deri vatization method was selected.5-(2-Hydroxyethyl) benzoacridine (HBA) reacts with coupling agent N,N'-carbonyldiimidazole(CDI) to form an activated amide intermediate 2-(benzoacridin) ethyl-imidazole-1-carbox-ylate(BAEIC).BAEIC, which is dual-sensitive probe, reacts preferably with amino compounds at 80 ℃ in the presence of 4-dimethylaminopyridine(DMAP) catalyst in N,N-dimethylformamide(DMF) solvent to give the corresponding sensitively fluorescent derivatives with an excitation maximum at λ_(ex) of 280 nm and an emis sion maximum at λ_(em) of 510 nm.BAEIC-amine derivatives simultaneously exhibited high ionization potential with percent ionization (changing from 5.62% to 58.08% in aqueous acetonitrile and from 2.14% to 56.58% in aqueous methanol.Derivatives were not only sensitive to fluorescence but also to MS ionizable potential.The fluorescence detection limits(5/iV = 3) were 0.12-0.59 μg/L.The online APCI-MS detection limits were 1.9-14 μg/L(S/N=5).

Objective To provide a theoretical foundation for language rehabilitation by investigating the characteristics of verb impairment in Chinese agrammatism(CA)and exploring its mechanism.Methods Fifteen CA patients and 15 normal subjects(control group)were recruited and evaluated using a standardized aphasia battery in Chinese,the Chinese agrammatism battery,a verb comprehension and production task and a short-term memory task.Results Compared with the control group,verb comprehension and production,as well as working memory were significantly damaged among the CA patients,and it was more difficult for the agrammatic aphasics to produce than to comprehend verbs(P＜0.01).Verbs with a more complex argument structure(in terms of the number and type of arguments)were more difficult for the agrammatie aphasics to produce than those with a less complex argu- ment structure.But the complexity of the argument structure did not influence the agrammatic aphasics'verb compre- hension.Conclusion The semantic entry of a verb,shared by both comprehension and production,was intact,but the verb's grammar entry was impaired.Poor working memory together with the complexity of the argument structure accounted for the impaired verb production of the agrammatic patients.