In order to establish the optimum condition for purifica tion of the nucleoprotein(NP) of rabies virus by immunoaffinity chromatography, the efficient and non-denaturative eluents(Mg-el) was obtained by using ELISA elution model; furthermore, it didn't damage the activity of NP. Two kind of NPs , expressed by recombinant vaccinia virus (rVac-N) and recombinant baculovirus (BRN), were purified by a Sepharose CL 4B column and a 2C12- Sepharose 4B colum n. By Western-blot and SDS-PAGE, high purity and good antigenical intact NPs w ere identified. The purified ribonucleoprotein (RNP) of rabies virus 5aG strain was also obtained. After immunized with NP and RNP, mice developed a strong anti -nucleoprotein response and were protected against a lethal challenge of rabies virus CVS strain. There were not difference been observed among the mice immuni zed with different purified protein. These data indicate that the NPs are antige nical and immunogenical comparable to the authentic rabies RNP and therefore pre sent a potential source of an effective ,safe and economical subunit vaccine.

The rice water weevil (RWW) Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae) is an invasive insect pest of rice Oryza sativa L. in China. Little is known about the interactions of this weevil with indigenous herbivores. In the present study, adult feeding and population density of the weevil, injury level of striped stem borer Chilo suppressalis (Walker) (Lepidoptera: Pyralidae) and pink stem borer Sesamia inferens (Walker) (Lepidoptera: Noctuidae) to rice, as well as growth status of their host plants were surveyed in a rice field located in Southeastern Zhejiang, China, in 2004 with the objective to discover interspecific interactions on the rice. At tillering stage, both adult feeding of the weevil and injury of the stem borers tended to occur on larger tillers (bearing 5 leaves) compared with small tillers (bearing 2~4 leaves), but the insects showed no evident competition with each other. At booting stage, the stem borers caused more withering/dead hearts and the weevil reached a higher density on the plants which had more productive tillers and larger root system; the number of weevils per tiller correlated negatively with the percentage of withering/dead hearts of plants in a hill. These observations indicate that interspecific interactions exist between the rice water weevil and the rice stem borers with negative relations occurring at booting or earlier developmental stages of rice.
Animals ; Coleoptera ; growth & development ; Oryza ; growth & development ; parasitology ; Population Density ; Weevils ; growth & development

OBJECTIVETo investigate the clinical value of matrix assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS) in detecting K-ras gene mutation.

METHODSSixty-one paraffin-embeded specimens of colorectal cancer were selected. MALDI-TOF-MS and regular sequencing were used to test the mutation of codon 12 and 13 in K-ras exon 2.

RESULTSOnly 47 specimens could be detected successfully in regular sequencing, while all the specimens were tested successfully in MALDI-TOF-MS. Fourteen specimens had K-ras mutation in regular sequencing (30.0%), while 22 specimens had mutation in MALDI-TOF-MS (36.1%). Six specimens with mutation were found in MALDI-TOF-MS but were wild-type in regular sequencing. Same mutation types from 14 specimens were confirmed by both regular sequencing and MALDI-TOF-MS. MALDI-TOF-MS was able to detect the mutation in 2 specimens that was not identified in regular sequencing.

CONCLUSIONSMALDI-TOF-MS is a feasible approach of K-ras gene mutation testing in colorectal cancer, which is less demanding in terms of specimen quality and is more sensitive as compared to regular sequencing.

Colorectal Neoplasms ; genetics ; Genes, ras ; Humans ; Mutation ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

BACKGROUNDWith economic growth and urbanization there have been significant changes in the life style and diet of urban residents in large cities of China, which is experiencing a rapid increase in the prevalence of diabetes. While high prevalence of diabetes has been reported, little is known of the long-term effects of diabetes in such a large population. The aim of this study was to estimate the morbidity rate of diabetic peripheral neuropathy (DPN) in a Chinese urban diabetic population with more than 10 years' disease duration, and evaluate the relevant risk factors. The clinical manifestation of DPN and pain status was also assessed.

METHODSFive hundred and sixty-five diabetes patients were recruited into the study. Symptoms and examination helped diagnose neuropathy. The clinical manifestation of DPN was assessed with a visual analog pain score (VAS). Diabetic complication status was determined from medical records. Serum lipids and lipoproteins, glycosylated hemoglobin (HbA1c), and the urinary albumin excretion rate were measured.

RESULTSThe morbidity rate of DPN was 46.6%. HbA1c, hyperlipidemia, and retinopathy were significantly associated with neuropathy, and these risk factors were correlated with other diabetic micro and/or macrovascular complications. The average VAS pain score of the DPN patients was 4.12 ± 2.07. Severe and moderate pain was experienced by 11.4% and 40.5% respectively of DPN patients. About 3.7% of diabetic subjects had lower limb ulcer or amputation.

CONCLUSIONSThe morbidity rate of DPN for diabetic patients with > 10 years duration is very high compared to the range reported for other populations in the world. The risk factors for DPN include HbA1c, hyperlipidemia, and retinopathy. In long-standing diabetic patients, DPN was not associated with diabetic duration, and half of the DPN patients experienced considerable daily suffering.

Aged ; China ; Diabetic Neuropathies ; epidemiology ; metabolism ; physiopathology ; Female ; Glycated Hemoglobin A ; metabolism ; Humans ; Hyperlipidemias ; epidemiology ; metabolism ; physiopathology ; Male ; Middle Aged ; Pain ; etiology ; Risk Factors ; Urban Population

OBJECTIVETo explore biological exposure markers, we investigated the chromium content in peripheral erythrocytes from occupational population with broad ranges of soluble chromate exposure, as the candidate biomarker may provide the scientific evidence for health risk assessment in occupational chromate-exposed population.

METHODSA cross-sectional study was conducted in chromate exposed workers employed at a chromate factory in a district of Jinan city, Shandong Province. The studied population contained 114 workers from different processes of the chromate plants, which included 74 males and 40 females, with an age range from 25 to 52 years old, averaging at (35.83 +/- 6.14) years old; the length of service was ranging from 1 year to 37 years, an average of (14.20 +/- 6.77) years. In addition, 30 farmers in the countryside one hundred kilometers away from the factory, without exposure to chromate matched with exposed subjects by age, gender and smoking status were identified as a control group, which included 22 men and 8 women, with age ranging from 25 years old to 47 years old, having an average age of (36.13 +/- 6.17) years old. Personal information on age, chromate exposure, medical history, smoking habit and alcohol consumption was obtained at an interview. The air concentration of personal exposure was determined by individual sampling for 8 hours per day as shift work, and chromium was assayed by atomic absorption spectrometry. The chromium content in the erythrocytes from peripheral blood was determined by graphite furnace atomic absorption spectrometry. The potential plasma reduction capacity was determined by dibenzene anthracoamid dihydrazide spectrophotometry. The content of total vitamin C and reductive ascorbic acid were determined by 2, 4-dinitrobenzene hydrazine. The data were analyzed by SPSS10.0 software for statistical significance.

RESULTS(1) The results showed that the chromium levels in erythrocytes in the exposed group [(15.79 +/- 31.01) microg/L] were significantly higher than those in the control group [(3.21 +/- 2.20) microg/L] (P < 0.01). (2) There existed a dose-response relationship between the personal airborne chromate concentration and the chromium content in erythrocytes. As airborne chromate concentration lowered to 106.00 microg/m(3), the chromium content in erythrocytes increased, depending on the air concentration of chromate. (3) Correlation analysis showed that there was a significant positive correlation between airborne chromate concentration and the chromium content in erythrocytes (P < 0.01). (4) In multiple regression analysis, it was found that the potential plasma reduction capacity and reductive ascorbic acid may be a good indicator for oxidative stress produced by chromate exposure and be used to evaluate the effects on intracellular uptake of chromium (VI).

CONCLUSIONOur findings suggested that the chromium content in erythrocytes should be used as an effective exposed biomarker in the risk assessment for occupational chromate-exposure.

Adult ; Air Pollutants, Occupational ; analysis ; Biomarkers ; blood ; Chromates ; analysis ; Chromium ; blood ; Erythrocytes ; chemistry ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; analysis

OBJECTIVETo observe the antileukemic effect of lymphocytes from cord blood treated by CpG-oligodeoxynucleotides (CpG-ODN).

METHODSLymphocytes from cord blood were exposed to different oligodeoxynucleotides containing a panel of CpG-ODN and were cultured with K562 cells. The cytotoxic effects were detected by MTT method. Immunological markers of cord blood treated by CpG-ODN(3) which showed highest activity were measured with flow cytometry.

RESULTSDifferent CpG-motifs have different immunostimulatory activity and CpG-ODN(3) has the highest one. After treated by CpG-ODN(3), NK killing activity to K562 cells increased in a dose-dependent manner, and CD(3), CD(4), CD(19) and CD(56) increased to (60.6 +/- 7.9)%, (40.2 +/- 3.5)%, (22.4 +/- 1.9)% and (15.5 +/- 3.1)%, respectively.

CONCLUSIONCpG-ODN could reinforces the immunological competence of cord blood lymphocytes and their effects on K562 cells. This provides a new approach to reinforce the antitumor effects of cord blood.

Adjuvants, Immunologic ; pharmacology ; Antigens, CD ; blood ; Cytotoxicity, Immunologic ; Dose-Response Relationship, Drug ; Fetal Blood ; cytology ; Humans ; K562 Cells ; Leukemia ; therapy ; Lymphocytes ; drug effects ; immunology ; Oligodeoxyribonucleotides ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction

Phytase gene of Aspergillus niger 963 was cloned into baculovirus transfer vector. DNA of the recombinant vector was co-transfected with Bm-BacPAK6 DNA into BmN cells, and recombinant virus was selected by plaque assays. The recombinant virus was identified by Dot blot and Southern-blot with the specific probe for phytase gene. Phytase gene was expressed in silkworm larvae and pupae. The expression product was 1.43 g/L haemolymph for silkworm larvae and 1.90 g/L haemolymph for pupae, respectively. The enzymic characteristicses of phytase expressed in baculovirus-expression system were studied in this paper.
6-Phytase ; genetics ; metabolism ; Animals ; Aspergillus niger ; enzymology ; genetics ; Blotting, Southern ; Bombyx ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Fungal Proteins ; genetics ; metabolism ; Genetic Vectors ; genetics ; Hydrogen-Ion Concentration

OBJECTIVETo establish a method for real-time recording the oxygen consumption of mice under normobaric hypoxia.

METHODSThe experimental apparatus was made up of animal container, filling water control system, electronic balance, hose, a computer with weight recording software, etc. The working principle was that the oxygen consumed by animal was replaced by water filling which was controlled by the pneumatic and hydraulic actuator. The water was weighted by an electronic balance and the weight signal was recorded into excel file at the same time. The accuracy and precision of the apparatus were detected by a 10 ml syringe. The oxygen consumption characteristics of 6 acute repetitive hypoxia mice and 6 normal mice were observed.

RESULTSThe P value for the paired t test was 1 and the CV value was 4%. The survival time and total oxygen consumption of acute repetitive hypoxia mice were both significantly increased compared to normal mice (P < 0.05), which were (58.8 +/- 6.8) min and (46.0 +/- 8.7) min respectively for the survival time and (85.1 +/- 8.5) ml and (73.6 +/- 5.4) ml respectively for total oxygen consumption.

CONCLUSIONThe hypoxia tolerance of the acute repetitive hypoxia mice can significantly improved by taking more oxygen in the animal cabin. The accuracy and precision of the apparatus are high and it can be used for the determination of oxygen consumption in hypoxia research.

Animals ; Hypoxia ; physiopathology ; Mice ; Monitoring, Physiologic ; instrumentation ; Oxygen Consumption ; physiology

Background Leber hereditary optic neuropathy (LHON)is a common inherited eye disease,which generally affects young adults with bilateral loss of central vision.Mutation frequency of Leber hereditary has not been fully clarified. Objective This study was to investigate the mutation frequency of mitochondrial NDI gene associated with LHON in Chinese population. Methods The proposal of the study was approved by Ethic Committee of Wenzhou Medical College.Written informed consent was obtained from each subject initial of this trial.Eight hundred and ninety-four LHON patients and 134 normal subjects were collected.Genomic DNA was extracted from peripheral blood leukocytes of the all participants.Polymerase chain reaction (PCR) was used to amplify and sequence analysis of the mitochondrial ND1 gene was performed and aligned with revised Cambridge Reference Sequence(rCRS) of mitochondrial DNA.Then mutated gene frequency was screened and analyzed. Results Mutational analysis of mitochondrial ND1 gene in 894 LHON patients revealed the presence of G3316A,T3394C,G3460A,C3497T,G3635A,G3733A,and T4216C.11.19％ LHON patients (100/894 ) were found to be associated with the gene mutations mentioned above,and 3.24％ patients (29/894) showed the co-occurrence of three primary mutations.Mutation frequencies in LHON patients were 2.57％,2.23％,1.45％,3.80％,0.67％,0.11％,0.34％,respectively,and G3316A,T3394C,C3497T and T4216C also were detected in 134 normal controls with the mutation frequencies of 4.48％,2.99％,4.48％ and 1.49％,respectively.Mutation frequency analysis showed an insignificant difference in the mutations of G3316A,T3394C,C3497T and T4216C between LHON patients and normal controls (x2 =0.926,P=0.336;x2 =0.052,P=0.820; x2 =0.142,P=0.707;P=0.129).G3376A,G3496T,G3700A,A4136G,T4160C and C4171A were absent in Chinese LHON patients. Conclusions Mitoehondrial ND1 gene in LHON is a mutational hotspot in Chinese population,11.19％ (100/894)associated with LHON was caused by ND1 gene mutation.G3635A,G3733A may be rare pathological mutation in Chinese population.However,G3316A,T3394C,C3497T and T4216C are insufficient to produce the clinical phenotype,but they may play a synergic role for penetrance and phenotypic manifestation in LHON.

Objective To determine the value and the optimal cutoff point of waist circumference (WC) in screening diabetes mellitus (DM) and to provide evidence for DM prevention and identifying population at risk in mid-western rural areas of Shandong province.Methods A sample consisting 16 341 rural residents was selected and studied.All participants were physically examined on height,weight,WC and fasting plasma glucose (FPG).Oral glucose tolerance test (OGTT) was performed for subjects with FPG valued from 6.1 to 7.0 mmol/L.DM was defined according to the criteria set by WHO in 1999.Area under the curve (AUC),sensitivity,specificity and Youden index were computed based on the receiver operating characteristic (ROC) curve analysis.Optimal cutoff point was determined by the maximum of Youden index.Results The prevalence rates of DM for males and females increased along with the rise of WC (trend test X2=72.01,122.65,P＜0.01 ).It appeared significantly higher in those with WC 85 cm in females and≥80 cm in males,with those WC ＜85 cm for females and ＜80 cm for males,in particular.AUCs were 0.639 and 0.655 for males and females respectively and both had significant differences (t=7.22,11.07,P ＜0.01 ).However,the AUCs did not show significant difference (t=0.70,P ＞ 0.05) between males and females.The Youden index reached maximum when WC approached 85 cm for females (24.90%) and 80 cm for males (24.39%).The sensitivity and specificity were 58.04%and 66.86%for males,and 67.08%and 57.31%for females.Conclusion WC seemed to be an effective indicator for screening the DM.The optimal cutoff point of WC would be 85 cm for females and 80 cm for males in screening DM and defining the population at risk in this area.