In order to prepare monoclonal antibody to σ A protein of avian reovirus(ARV)and es-tablish a sandwich ELISA method for the detection of ARV pathogens.In this study,the σ A pro-tein of ARV was expressed as antigen by prokaryotic expression and used to immunize BALB/c mice.Then,stable hybridoma cell lines were screened,and monoclonal antibodies were prepared.A sandwich ELISA detection method based on monoclonal antibody of σA protein was established,and the sensitivity,specificity,repeatability,and accuracy were tested.The results showed that the recombinant plasmid pET-32a-σA was successfully constructed and well expressed in Escherichia coli.After immunizing mice,two hybridoma cell lines 6B3 and 8E11,which could secrete mono-clonal antibodies stably,were successfully prepared.Both monoclonal antibodies could react with natural ARV.One of the monoclonal antibodies secreted by 6B3 was selected as the capture anti-body and the ARV-positive chicken polyclonal antibody was used as the detection antibody.A sand-wich ELISA method was established to detect ARV by optimizing the reaction conditions.The specific test showed that the method only detected ARV pathogens and no other common chicken viral pathogens were detected.The detection limit was 7.72 X 102 EID50/mL of ARV antigen.The coefficient of variation of the intra-and inter-assay tests were less than 5.0％and the reproducibili-ty was good.Thirty samples were tested simultaneously by σA-sandwich ELISA and PCR,and the results were consistent with each other.In conclusion,a sandwich ELISA method based on the monoclonal antibody of σA protein was successfully established for the identification and detection of ARV,which provided a technical means for the accurate and rapid detection of ARV.

The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β₂m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a Superdex^TM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.
Animals ; Capripoxvirus ; Epitopes, T-Lymphocyte/genetics* ; Peptides/genetics* ; Poxviridae Infections ; Sheep ; Sheep Diseases

Objective: To characterize the clinical epidemiological features of primary lung cancer patients based on massive clinical data. Methods: The demographic and histological information of 8 081 primary lung cancer patients who were initially identified from 1 January 2012 to 31 December 2013 in 16 hospitals from 6 provinces were retrospectively analyzed to determine the characteristics of different histological subtypes among different gender, age-group and birth cohort. Results: Among the 8 081 lung cancer patients, 5 422 (67.10%) were male and 2 659 (32.90%) were female, the average age was (60.96±10.14) years. The most common histological subtypes of lung cancer successively were adenocarcinoma (ADC), squamous cell carcinoma (SCC), small cell carcinoma and large cell carcinoma, and the proportions of these subtypes were 53.13%, 24.51%, 14.59% and 0.66%, respectively, which collectively accounted for 92.89%. The current highest smoking rate was observed in SCC patients, which was 62.45%, while lowest in ADC, which was 29.68%. The incidence of lung cancer in male was significantly higher that that of female, with a sex ratio of 2.04∶1. The highest sex ratio was observed in SCC, which was 9.14∶1, while lowest in ADC, which was 1.14∶1. The distribution of histological subtypes in male lung cancer patients was consistent with the general situation.While among the female lung cancer patients, the proportion of ADC was the highest (75.42%), followed by SCC (10.08%), squamous cell carcinoma (7.34%) and large cell carcinoma (0.39%). Analyzed by the birth cohort, the proportion of ADC gradually increased with the age, while the reduced tendency was observed in SCC (P<0.000 1). Conclusion Adenocarcinoma is the most important histological subtype of lung cancer, and the distribution characteristics of histological subtypes of lung cancer differs among genders, age and birth cohort.

OBJECTIVE: To explore the role of transforming growth factor-β1/integrin-linked kinase/fibroblast-specific protein 1 (TGF- β1/ILK/FSP1) signaling pathway in cyclosporine A (CsA)-induced renal tubular epithelial cell transdifferentiation. METHODS: Rat renal tubular epithelial NRK-52E cells were induced with 1 mg/L CsA, treated with TGF-β1 inhibitor (SB431542, 10 μmol/L), or transfected with the ILK-RNAi lentiviral expression vector (ILKshRNA) or a negative control vector before CsA induction. The expressions of TGF-β1, ILK and FSP-1 mRNAs and proteins in the cells were detected using real-time PCR and Western blotting. The positive cells for α-SMA expression were detected by immunohistochemistry. RESULTS: Compared with the blank control cells, the cells treated with CsA showed significantly increased levels of TGF-β1, ILK and FSP-1 mRNAs and proteins ( < 0.05). The expressions of TGF-β1, ILK and FSP-1 were significantly lower in TGF-β1 inhibitor group than in CsA group ( < 0.05). The levels of ILK and FSP-1 were significantly decreased after shRNA-mediated ILK silencing ( < 0.05). The number of positive cells for -SMA was significantly lower in cells treated with SB431542 and in cells with ILK silencing than in the cells treated with CsA alone ( < 0.05). CONCLUSIONS The activation of TGF-β1/ILK/FSP-1 signaling pathway is an important mechanism for CsA-induced transdifferentiation in rat renal tubular epithelial cells. ILK participates in CsA-induced epithelialmesenchymal transition of renal tubular epithelial cells.
Animals ; Calcium-Binding Proteins ; Cells, Cultured ; Cyclosporine ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; Protein-Serine-Threonine Kinases ; Rats ; Signal Transduction ; Transforming Growth Factor beta1

Objective To characterize the clinical epidemiological features of primary lung cancer patients based on massive clinical data. Methods The demographic and histological information of 8 081 primary lung cancer patients who were initially identified from 1 January 2012 to 31 December 2013 in 16 hospitals from 6 provinces were retrospectively analyzed to determine the characteristics of different histological subtypes among different gender, age?group and birth cohort. Results Among the 8 081 lung cancer patients, 5 422 (67.10%) were male and 2 659 (32.90%) were female, the average age was (60.96± 10.14) years. The most common histological subtypes of lung cancer successively were adenocarcinoma ( ADC ), squamous cell carcinoma ( SCC ), small cell carcinoma and large cell carcinoma, and the proportions of these subtypes were 53.13%, 24.51%, 14.59% and 0.66%, respectively, which collectively accounted for 92.89%. The current highest smoking rate was observed in SCC patients, which was 62.45%, while lowest in ADC, which was 29.68%. The incidence of lung cancer in male was significantly higher that that of female, with a sex ratio of 2.04 ∶ 1. The highest sex ratio was observed in SCC, which was 9.14 ∶ 1, while lowest in ADC, which was 1.14 ∶ 1. The distribution of histological subtypes in male lung cancer patients was consistent with the general situation.While among the female lung cancer patients, the proportion of ADC was the highest (75.42%), followed by SCC (10.08%), squamous cell carcinoma ( 7.34%) and large cell carcinoma (0.39%). Analyzed by the birth cohort, the proportion of ADC gradually increased with the age, while the reduced tendency was observed in SCC ( P＜0.000 1). Conclusion Adenocarcinoma is the most important histological subtype of lung cancer, and the distribution characteristics of histological subtypes of lung cancer differs among genders, age and birth cohort.

Objective To characterize the clinical epidemiological features of primary lung cancer patients based on massive clinical data. Methods The demographic and histological information of 8 081 primary lung cancer patients who were initially identified from 1 January 2012 to 31 December 2013 in 16 hospitals from 6 provinces were retrospectively analyzed to determine the characteristics of different histological subtypes among different gender, age?group and birth cohort. Results Among the 8 081 lung cancer patients, 5 422 (67.10%) were male and 2 659 (32.90%) were female, the average age was (60.96± 10.14) years. The most common histological subtypes of lung cancer successively were adenocarcinoma ( ADC ), squamous cell carcinoma ( SCC ), small cell carcinoma and large cell carcinoma, and the proportions of these subtypes were 53.13%, 24.51%, 14.59% and 0.66%, respectively, which collectively accounted for 92.89%. The current highest smoking rate was observed in SCC patients, which was 62.45%, while lowest in ADC, which was 29.68%. The incidence of lung cancer in male was significantly higher that that of female, with a sex ratio of 2.04 ∶ 1. The highest sex ratio was observed in SCC, which was 9.14 ∶ 1, while lowest in ADC, which was 1.14 ∶ 1. The distribution of histological subtypes in male lung cancer patients was consistent with the general situation.While among the female lung cancer patients, the proportion of ADC was the highest (75.42%), followed by SCC (10.08%), squamous cell carcinoma ( 7.34%) and large cell carcinoma (0.39%). Analyzed by the birth cohort, the proportion of ADC gradually increased with the age, while the reduced tendency was observed in SCC ( P＜0.000 1). Conclusion Adenocarcinoma is the most important histological subtype of lung cancer, and the distribution characteristics of histological subtypes of lung cancer differs among genders, age and birth cohort.

Objective: To estimate the incidence and mortality of female breast cancer in China based on the cancer registration data in 2014, collected by the National Central Cancer Registry (NCCR), and to provide support data for breast cancer prevention and control in China. Methods: There were 449 cancer registries submitting female breast cancer incidence and deaths data occurred in 2014 to NCCR. After evaluating the data quality, 339 registries′ data were accepted for analysis and stratified by areas (urban/rural) and age group. Combined with data on national population in 2014, the nationwide incidence and mortality of female breast cancer were estimated. Chinese population census in 2000 and Segi′s population were used for age-standardized incidence/mortality rates. Results: Qualified 339 cancer registries covered a total of 288 243 347 populations (144 061 915 in urban and 144 181 432 in rural areas) in 2014. The morphology verified cases (MV%) accounted for 87.42% and 0.59% of incident cases were identified through death certifications only (DCO%), with mortality to incidence ratio of 0.24. The estimates of new breast cancer cases were about 278 900 in China in 2014, accounting for 16.51% of all new cases in female. The crude incidence rate, age-standardized rate of incidence by Chinese standard population (ASRIC), and age-standardized rate of incidence by world standard population (ASRIW) of breast cancer were 41.82/100 000, 30.69/100 000, and 28.77/100 000, respectively, with a cumulative incidence rate (0-74 age years old) of 3.12%. The crude incidence rates and ASRIC in urban areas were 49.94 per 100 000 and 34.85 per 100 000, respectively, whereas those were 31.72 per 100 000 and 24.89 per 100 000 in rural areas. The estimates of breast cancer deaths were about 66 000 in China in 2014, accounting for 7.82% of all the cancer-related deaths in female. The crude mortality rate, age-standardized rate of mortality by Chinese standard population(ASRMC) and age-standardized rate of mortality by world standard population (ASRMW) of breast cancer were 9.90/100 000, 6.53/100 000, and 6.35/100 000, respectively, with a cumulative mortality rate of 0.69%. The crude mortality rates and ASRMC in urban areas were 11.48 per 100 000 and 7.04 per 100 000, respectively, whereas those were 7.93 per 100 000 and 5.79 per 100 000 in rural areas. The incidence and mortality rates of breast cancer were higher in areas than those in rural areas. The age-specific incidence rates of breast cancer increased greatly after 20 years old and peaked at the age group of 55-60. The age-specific mortality rates increased rapidly with age, particularly after 25 years old. They remained at a relative stable level from 55 to 65 years of age, and then increased dramatically and peaked in the age group of 85 and above. Conclusions Breast cancer is still one of the most common malignant tumor threatening to famale health in China. The disease is more prevalent in urban areas at the age group of 55-60. Comprehensive prevention and control strategies referring to local status and age groups should be carried out to reduce the burden of breast cancer.

Objective: To estimate the incidence trend and change in the age distribution of female breast cancer in cancer registry areas in China from 2000 to 2014. Methods: 22 cancer registries in China with continuous monitoring data from 2000 to 2014 were selected. All datasets were checked and evaluated based on data quality control criteria and were included in the analysis. The cancer registries covered 675 954 193 person-years, including 342 010 930 person-years of male and 333 943 263 person-years of female. Female breast cancer cases (International Classification of Diseases-10^th Revision: C50) were extracted. Crude incidence rate (CR), age-standardized incidence rate by Chinese standard population(ASIRC), annual percent change (APC), crude and adjusted mean age at onset were calculated. Incidence rates stratified by regions and age groups were calculated. Results: Female breast cancer incidence rate significantly increased from 31.90/100 000 in 2000 to 63.30/100 000 in 2014. Incidence rate increased rapidly from 2000 to 2008 (CR: APC=6.5%, 95%CI: 5.3%-7.8%; ASIRC: APC=4.6%, 95%CI: 3.6%-5.7%). Its increment slowed down from 2008-2014 (CR: APC=3.2%, 95%CI: 1.4%-5.1%; ASIRC: APC=1.4%, 95%CI:-0.1%-2.9%). The crude mean age at onset increased from 54.4 in 2000 to 57.0 in 2014. Adjusted mean age at onset remained around 54.3 in 2014. Crude mean age at onset increased significantly over time in all registry areas (β=0.192, P<0.001), urban (β=0.205, P<0.001) and rural (β=0.092, P=0.014) areas, while adjusted mean age at onset remained stable in all registry areas (β=0.009, P=0.289), urban (β=0.017, P=0.139) and rural (β=-0.054, P=0.109) areas. Conclusion Female breast cancer incidence rate in China increased from 2000 to 2014. Aging of the population resulted in a significant increase in crude mean age at onset. After age adjustment, no significant changes in age distribution were found.

Objective: To analyze the incidence trend and mean age at diagnosis for lung cancer in cancer registration areas of China from 2000 to 2014. Methods: The data of lung cancer incidence used in this study were from 22 registries submitted to National Central Cancer Registry with continuous data during 2000 and 2014, covering about 621 593 469 person-years. All cancer cases were coded as C33-C34 according to the International Classification of Diseases-10^th Revision (ICD-10) were extracted for this analysis with about 343 663 patients. The incidence of different sex and regional population, the standardized incidence rate by Chinese population, the average annual change percentage (AAPC), the mean age and adjusted mean age of cancer incidence were calculated. The incidence of each year was described by regional and age groups, and the linear regression model was employed to analyze the relationship between mean age at onset and year. Results: The crude incidence rate and age-standardized incidence rate (ASR) of lung cancer for men in cancer registry areas in 2000 were 56.98 per 100 000 and 48.43 per 100 000, respectively. The rates were 89.51 per 100 000 and 46.85 per 100 000 in 2014, respectively. For women in the same areas, the rates were 27.77 per 100 000 and 20.17 per 100 000 in 2000; while 51.31 per 100 000 and 25.44 per 100 000 in 2014, respectively. The crude incidence rate increased along with the age. In 2000-2014, the trend of crude rate and ASR of lung cancer were significantly increased (CR: AAPC=3.8%, 95%CI: 3.5%-4.1%; ASR: AAPC=0.4%, 95%CI: 0.2%-0.7%). The rise of crude rate in females was higher than that in males (Male: AAPC=3.5%, 95%CI: 3.2%-3.7%; Female: AAPC=4.5%, 95%CI: 4.1%-5.0%). However, the rise of the ASR declined for both male and female (Male: AAPC=-0.2%, 95%CI:-0.4%-0.0%; Female: AAPC=1.4%, 95%CI: 1.0%-1.9%). The average age at diagnosis of lung cancer in rural areas was 64.35 years old in 2000, and increased to 65.97 years old in 2014 (β=0.11, P<0.001), while adjusted mean age at onset remained stable in all areas and urban areas (P>0.05). And the average age at onset increased significantly over time in male (β=-0.02, P=0.014), which was not seen in female (β=-0.01, P=0.522). Conclusion The crude incidence rate of lung cancer in cancer registry areas in China increased slowly during 2000-2014; and the standardized average age of male at diagnosis decreased slightly, while the age in rural areas increased during 2000-2014. Lung cancer will still be the focus of cancer prevention and control in the near future.

Objective: To analyze the trend of cancer incidence and age changes among men in cancer registration areas of China from 2000 and 2014. Methods: We select the information of national cancer registry with continuous data from 2000 to 2014, review and organize the monitoring data at the above registries. A total of 22 monitoring registries were included in this study. The covering population of male were about 314 330 648 person years. The information on the incidence of all male prostate cancer patients with C61 was extracted from the International Classification of Diseases-10^th Revision (ICD-10). To understand the incidence of male prostate cancer in each year, the age-standardized rate by Chinese population (ASR), average annual percent change (AAPC), adjusted mean age at onset were calculated. Incidence rates stratified by regions and age groups were also calculated. The linear regression model was employed to analyze the relationship between mean age at onset and year. Results: The prostate cancer incidence in China increased by 11.5% (95%CI: 10.3%-12.7%) from 2000(4.62/100 000) to 2014(21.62/100 000), the age-standardized incidence rate increased by 7.1% (95%CI: 6.0%-8.1%) and the growth of rural was greater than that of urban. The age-specific incidence showed that the incidence rate increased significantly among the age group of 50 years; the incidence rates in men who have the same age but with different birth years showed a significant increase as birth years increased. The adjusted mean age at diagnosis of prostate cancer in cancer registry areas was 74.09 years old in the year of 2000, reduced by 0.13 year old to 72.35 years old in 2014 (β=-0.13, P<0.001). The adjusted mean age at onset declined significantly over time in urban areas (β=-0.13, P<0.001). Conclusion The trend of prostate cancer incidence among men in cancer registry regions generally increased, and the average age at diagnosis declined slightly from 2000 to 2014.