Congenital Abnormalities ; epidemiology ; Hospitals ; statistics & numerical data ; Humans ; Population Surveillance ; methods

OBJECTIVETo develop the monoclonal antibody against N protein of SARS virus and study its applicability.

METHODSBALB/c mice were immunized with recombinant N protein. Spleen cells were collected and infused with SP2/0 cell. The infused cells were screened for anti-N protein antibody with ELISA. The positive cells were cloned and injected into abdominal cavity. The antibodies were purified from ascites. The affinities of those purified antibodies were analyzed with ELISA. The ELISA for detection of SARS virus antigen was developed by using antibody with the highest affinity. Its sensitivity and specificity were also evaluated primarily.

RESULTSEleven monoclonal cells secreting antibody have been developed. Three of the 11 purified monoclonal antibodies had very high affinity to N protein, while 4 purified McAbs showed very weak reaction to N protein, the affinities of remaining 4 McAbs were in between. The ELISA for detection of SARS virus antigen was developed with McAb 7. Its sensitivity was about 31 PFU/ml and had no cross reaction with other respiratory viruses.

CONCLUSIONThe monoclonal antibody has good specificity and may be used to detect SARS virus antigen. However, its sensitivity is to be evaluated further with clinical samples from SARS patients, especially at acute phase.

Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibodies, Viral ; biosynthesis ; Antigens, Viral ; analysis ; Humans ; Hybridomas ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; immunology ; SARS Virus ; immunology ; Sensitivity and Specificity

OBJECTIVETo observe the therapeutic effect of Qingfei Huatan Quyu method (QHQ, a Chinese medicinal therapy for clearing Fei-heat and dissolving phlegm-stasis) combined with hormone-antibiotic therapy (HAT) on radiation pneumonia (RP).

METHODSEighty-one patients with RP were randomized into two groups, 41 patients in the control group and 40 in the treatment group were treated with HAT alone and HAT combined with QHQ respectively for 21 days. The severity of RP was evaluated before and after treatment according to the criteria of the radiation therapy oncology group. The effect on TCM symptoms and chest roentgenogram, as well as on plasma levels of interleukin-6 ( IL-6) and transform growth factor-beta (TGF-beta) were detected.

RESULTSAfter treatment, number of patients with RP graded as 0, 1, 2, 3, and 4 in the treatment group was 23, 10, 4, 2, and 1, respectively, while in the control group, 14, 9, 11, 4, and 3, respectively. The combined therapy showed effects in improving RP grading (P <0.01) and TCM syndromes were superior to those of HAT respectively (P < 0.05). Besides, levels of IL-6 and TGF-beta were lowered after treatment in the treatment group, showing a significant difference to those in the control group (P <0.05).

CONCLUSIONQHQ combined with HAT has a definite therapeutic effect on RP. It could efficiently decrease the plasma levels of IL-6 and TGF-beta in patients with RP.

Anti-Bacterial Agents ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Interleukin-6 ; blood ; Medicine, Chinese Traditional ; Radiation Pneumonitis ; drug therapy ; Transforming Growth Factor beta ; blood

A simple,rapid and sensitive upconversion immunochromatographic assay(UICA) was developed to detect imidaclothiz using NaYF4:Yb,Er upconversion nanoparticles(UCNPs) labeled with anti-imidaclothiz monoclonal antibody. The amino-modified UCNPs were conjugated with anti-imidaclothiz monoclonal antibody to prepare the UICA strip,which could realize the quantitative detection of imidaclothiz using a fluorescence photometer with an external 980 nm laser source. The working conditions of the UICA were systematically optimized, and the sensitivity, specificity, precision and accuracy were assessed by the studies of cross-reactivity (CR), spiked recovery and validation with HPLC. Under the optimal conditions (pH 8. 0, 0.3 mol/L NaCl,2.5% methanol and 0.2% PEG2000), the UICA could be completed in 25 min for the detection of imidaclothiz. The half-maximal inhibition concentration (IC50), limit of detection (IC10) and linear range (IC10-IC90) were 97.37 ng/mL,26.30 ng/mL and 26.30-363.08 ng/mL, respectively. The UICA had no CR with the analogues of imidaclothiz except for imidacloprid. The average spiked recoveries were 71.8%-97.2% with the relative standard deviations of 0.7%-10.7% in the matrices of paddy water, soil,pear,peach,wheat,cucumber,tomato and rice. The detection results of UICA for the authentic paddy water and pear samples were consistent with that of high performance liquid chromatography (HPLC).

OBJECTIVETo establish a national reference panel for HIV RNA diagnostic reagents.

METHODSSera from patients with HIV infection and healthy blood donors were collected and tested for HIV and HCV antibodies and HBsAg by using ELISA. The HIV antibody positive samples with ELISA were confirmed with HIV Blot 2.2 (Genelabs). The quantitative samples for HIV RNA were calibrated with the WHO HIV RNA standard. The stability of the panel was evaluated with acceleration method.

RESULTSAfter screening and calibration, 8 negative samples, 8 positive samples, 3 quantitative samples, 6 sensitivity samples and 5 samples for linear analysis were composed of the national reference panel for HIV RNA. The convinced international units (IU) for the quantitative samples were obtained by seven independent calibration and the logarithm of international units for the quantitative samples (b1-b3) were less than x +/- s. The results showed that this panel may stabilize for 4 days at 4 degrees C.

CONCLUSIONA national reference panel for HIV RNA reagents has been established. It may provide the basis for evaluating HIV RNA diagnostic reagents.

Blood Donors ; Calibration ; Drug Stability ; HIV Antibodies ; blood ; HIV Infections ; blood ; virology ; HIV-1 ; genetics ; isolation & purification ; Hepatitis B Surface Antigens ; blood ; Hepatitis C Antibodies ; blood ; Humans ; Indicators and Reagents ; standards ; RNA, Viral ; standards ; Reference Standards ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo study HIV, HBV and HCV infections in intravenous drug users.

METHODS2025 blood samples from intravenous drug users were collected from Sichuan, Hunan, Guangxi and Xinjiang regions, and tested for anti-HIV, anti-HCV, HBsAg using enzyme-linked immuno-sobent assays (ELISAs).

RESULTSThe positive rates of anti-HIV,anti-HCV and HBsAg were14.7%-30.4%, 60.7%-85.5% and 6.6%-22.4% in the intravenous drug users, respectively. The co-infection rates of HIV/HBV, HIV/HCV, HCV/HBV and HIV/HCV/HBV were 0%-0.4%, 11.6%-27.2%, 2.3%-14.3% and 1.6%-4.8% respectively in this population.

CONCLUSIONThe infection rates of HIV, HBV and HCV were higher in the intravenous drug users than that in general populations in the same regions, and HIV/HCV co-infection appeared most frequent in this population.

China ; epidemiology ; HIV Infections ; epidemiology ; Hepatitis B ; epidemiology ; Hepatitis C ; epidemiology ; Humans ; Prevalence ; Substance Abuse, Intravenous

Neoadjuvant chemoradiotherapy followed by surgery is the standard treatment for patients with locally advanced rectal cancer. Controversy on whether patients should receive radical surgery after pathological complete response (pCR) after neoadjuvant chemoradiotherapy has remained since pCR patients have shown favorable long-term outcome. Progress in multidisciplinary modalities has been made, including MRI, PET/CT imaging studies, genetic expression profiling, etc. The methods of predicting pCR response are inspiring. In this article, we review the methods for prediction and prognostic effect of pCR response when patients with locally advanced rectal cancer are treated with neoadjuvant chemoradiotherapy.
Chemoradiotherapy ; Humans ; Neoadjuvant Therapy ; Rectal Neoplasms ; therapy ; Remission Induction ; Treatment Outcome

OBJECTIVETo study the strategy of applying molecular genetic methods and techniques in the diagnosis of spinocerebellar ataxias (SCA).

METHODSThis study included 43 patients with SCA from 36 families, 38 sporadic SCA patients, 60 healthy individuals from the SCA families and 44 normal controls. The trinucleotide repeats were detected by polymerase chain reaction (PCR), denaturing polyacrylamide gel electrophoresis and silver staining technique. The repeat numbers were calculated by software.

RESULTSSCA3 was the most common type in the Hans of south China, accounting for 42.0%, followed by SCA2 (7.4%), SCA1 (4.9%), SCA7 (3.7%), SCA6 (2.5%) and SCA12 (1.2%). No patient was found to have SCA8, SCA10, SCA17, and dentatorubro-pallidoluysian atrophy(DRPLA).

CONCLUSIONMolecular genetic detection is an effective way to confirmation of SCA subtype diagnosis and presymptomatic genetic diagnosis.

Adult ; Electrophoresis, Polyacrylamide Gel ; Female ; Humans ; Male ; Middle Aged ; Pedigree ; Polymerase Chain Reaction ; Spinocerebellar Ataxias ; diagnosis ; genetics ; Trinucleotide Repeats ; genetics

OBJECTIVETo investigate the effects of lentivirus-mediated RNA interference targeting HOXA10 gene on the proliferation, apoptosis and morphology of leukemic cell line U937.

METHODSFour different shRNA plasmids were designed and built to interfere with HOXA10 gene. The four interference plasmids were transfected into 293T cells with the HOXA10 over expression plasmid and then the RNAi efficiency of the four interference plasmids was determined by Western blot. The best one was chosen to transfect 293T cells with lentiviral helping plasmids to produce packaged lentivirus (lenti-shHOXA10). U937 cells were divided into interference group (lenti-shHOXA10), negative control group and untreated group. After infection with the packaged lentivirus, infection efficiency of lentivirus for U937 was detected by flow cytometry, and the expression of HOXA10 gene mRNA and protein was detected by real-time PCR and Western blot. Cell survival was determined by MTT assay. Apoptosis rate was detected by flow cytometry.

RESULTSLentiviral-shRNA vector of HOXA10 gene was successfully constructed. Compared with the negative control and untreated groups, mRNA level of HOXA10 decreased by (92.3±1.3)%, protein levels decreased by 91.1%, and the inhibition rate of U937 cells [(43.9±0.7)%] increased in the interference group (P<0.05). Wright's staining showed that the ratio of karyon to cytoplasm was reduced and mitotic phase was rare in the interference group. Apoptosis rate in the interference group [(27.1±1.4)%] was significantly higher than in the negative [(19.4±1.9)%] and untreated groups [(5.5±1.3)%] (P<0.05).

CONCLUSIONSLentivirus mediated RNAi can reduce the expression level of HOXA10, effectively inhibit proliferation and promote apoptosis of U937 cells. HOXA10 gene is expected to become a new target for the treatment of leukemia at gene level.

Apoptosis ; Cell Proliferation ; Gene Silencing ; Homeodomain Proteins ; antagonists & inhibitors ; genetics ; Humans ; Lentivirus ; genetics ; RNA Interference ; Sequence Analysis, DNA ; U937 Cells

BACKGROUNDTo investigate the sensitivity and specificity of Procleix HIV/HCV RNA diagnostic assay.

METHODSHIV antibody positive or suspected positive plasmas of blood donors were collected from different provinces and detected with HIV antibody ELISA and HCV antibody ELISA. Samples positive for HIV by ELISA were confirmed by using HIV Blot. All the plasma samples were detected with Procleix HIV/HCV assay, HIV-1 discriminatory assay and HCV discriminatory assay, respectively.

RESULTSAll 74 samples positive for both HIV and HCV antibody were positive and 5 samples negative for both HIV and HCV antibody were negative when detected using Procleix HIV/HCV assay; 82 of 84 supplemental HIV antibody positive samples and 6 of 12 supplemental indeterminate samples were positive for HIV RNA, and all 7 HIV antibody negative samples were negative for HIV RNA when detected by using Procleix HIV discriminatory assay. Seventy of 81 HCV antibody positive samples and 4 of 22 HCV antibody negative samples were positive for HCV RNA when detected by using Procleix HCV discriminatory assay.

CONCLUSIONThis reagent is more sensitive and could be used in blood screening, thereby can reduce both HIV and HCV transmission of blood in window period of HIV and HCV infection.

Blood Donors ; HIV Infections ; diagnosis ; prevention & control ; virology ; HIV-1 ; genetics ; Hepacivirus ; genetics ; Hepatitis C ; diagnosis ; prevention & control ; virology ; Humans ; Nucleic Acid Amplification Techniques ; instrumentation ; methods ; RNA, Viral ; blood ; genetics ; Reproducibility of Results ; Sensitivity and Specificity