OBJECTIVETo establish and evaluate a BA-ELISA method for the quantitative detection of cystic fibrosis transmembrane conductance regulator (CFTR) protein.

METHODSWe deliberately selected three tables of CFTR and made the synthetic peptide be expressed in E. coli, then used the antigen to immunize rabbits to obtain the anti-CFTR polyclonal serum. After that, 96 well plates were coated with the purified antibody against CFTR. The antigen CFTR which was extracted from human sperm was detected by anti-CFTR antibody labeled with biotin, horseradish peroxidase conjugated avidin, and the substrate. The concentrations of two kinds of antibodies and the experiment parameters were optimized. Thereby, the double antibody sandwich BA-ELISA method for the quantitative detection of CFTR protein was established. Furthermore, the reproducibility, specificity and so on were evaluated by clinical specimens of sperm.

RESULTSThe optimal concentration of coated anti-CFTR IgG was 4 µg/ml, while the biotin labeled anti-CFTR IgG was 10 µg/ml; the optimal blocking buffer was 1% BSA-PBST, the optimal time of the reaction between antigen and antibody was 60 min, the optimal chromogenic time was 15 min, the intra-assay and inter-assay coefficient were 2.16%-9.23% and 2.29%-11.71% respectively; The lowest detectable limit was 0.15 ng/ml; the standard curve had a good linear correlation of R2 = 0.962.

CONCLUSIONThe BA-ELISA method for the quantitative detection of CTFR protein is successfully established, and it is demonstrated that the method has strong specificity, high sensitivity and good reproducibility. It provides the basis and evidence of the further application of the method.

Animals ; Antibodies ; Cystic Fibrosis Transmembrane Conductance Regulator ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; Humans ; Peptides ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity

Objective To evaluate the performance of magnetic nanoparticle chemiluminescence immunoassay (NM-CLIA) in detection of allergen-specific immunoglobulin E (sIgE) antibodies to Dermatophagoides pteronyssinus (International Allergen Code D1) and Dermatophagoides farinae (International Allergen Code D2). Methods A total of 489 serum samples from the patients with suspected allergic disease (244 cases caused by D1, and 245 caused by D2), who were treated at Suzhou Kowloon Hospital, Shanghai Jiaotong University School of Medicine, were detected by NM-CLIA and immunofluorescence assay, respectively. χ2 test and Kappa test were used to evaluate the correlation between the two methods in detection of D1 and D2 sIgE antibodies. The limit of detection (LoD), linear range and precision of NM-CLIA in detection of D1 and D2 sIgE antibodies were verified by the standard method of American Clinical Laboratory Standardization Association. Results The LoDs of NM-CLIA in detecting D1 and D2 sIgE antibodies were both less than 0.01 U/mL, the linearity ranged from 0.1 to 100 U/mL, the within-run precision was less than 5%, and the between-run precision was less than 8%. Methodological comparison results showed that NM-CLIA and immunofluorescence assay had good consistency in detecting D1 and D2 sIgE antibodies. For D1, the positive coincidence rate and negative coincidence rate were 95% and 92%, respectively (χ2＝174.45, P0.001, Kappa＝0.843), and the ±1 class agreement was 95.6%; for D2, the positive coincidence rate and negative coincidence rate were 91% and 97%, respectively (χ2＝154.263，P0.001，Kappa＝0.787), and the ±1 class agreement was 94.2%. Conclusion NM-CLIA has good correlation with immunofluorescence assay in detecting D1 and D2 sIgE antibodies, and has good LoD, linear range and precision, suggesting that it can be recommended for clinical testing of D1 and D2 sIgE antibodies.

Objective To investigate the prevalence of dyslipidemia and the relationship between breakfast frequency and dyslipidemia in residents over 40 years old in Yiling area of Yichang City in Hubei Province. Methods A random sampling was conducted, and 10 420 inhabitants were investigated during 2011 to 2012. Results The morbidity of dyslipidemia was 64. 0%. It was significantly higher in female than in male (65. 9% vs 60. 6%). Compared with regular breakfast eaters, non-breakfast eaters had significantly higher morbidity of higher blood low density lipoprotein-cholesterol(LDL-C) and hypertriglyceridemia(P<0. 05). The risk of elevated serum LDL-C was higher in the non-breakfast eaters group(OR=2. 382, 95%CI 1. 300-4. 367, P=0. 019) after adjusted by age, sex, smoking, drinking, etc. Conclusions Compared with regular breakfast eaters, breakfast skippers had significantly higher morbidity of dyslipidemia. Eating breakfast on daily basis may have a significant protective effect on preventing dyslipidemia.

OBJECTIVECancer is one of the leading causes of death in China. The study aimed to examine the temporal trend of cancer mortality rate during 1989-2008 in urban and rural areas of China.

METHODSThe mortality data of all cancers from 1989 to 2008 from National Cancer Registry database were sorted and checked. Age standardized mortality rates were calculated by the direct methods using the China population of 1982 and World Segi's population. Joinpoint regression was performed to obtain the annual percentage changes (APC) in mortality rates. The top ten cancer sites were calculated and analyzed. The mortality rates were compared with statistics of the United States.

RESULTSFrom 1989 to 2008, the trend of crude cancer mortality increased with an annual percentage change (APC) of 1.0%. After age standardization, the mortality rate was significantly decreased, with an APC of -1.2%. In urban areas, lung cancer was the most common cancer of death, whereas in rural areas, stomach cancer and esophageal cancer remained top cancers of death. Especially, in both urban and rural areas, the mortality of lung cancer was on increase. The mortality rates of stomach and esophageal cancers showed a decrease in urban areas. Compared with the cancer mortality rates of the United States, the Chinese cancer mortality rate in males remained highest. The decreasing trend of cancer mortality in females of China was less obvious than that of the United States.

CONCLUSIONSThe crude mortality rates of cancer in China show an increase whereas the age standardized mortality raters has declined between 1989 and 2008. Cancer is still a major public health issue threatening people's life in China. Effective intervention for cancer control and prevention is needed in the future.

China ; epidemiology ; Esophageal Neoplasms ; mortality ; Female ; Humans ; Lung Neoplasms ; mortality ; Male ; Mortality ; trends ; Neoplasms ; mortality ; Registries ; Rural Population ; Sex Factors ; Stomach Neoplasms ; mortality ; United States ; epidemiology ; Urban Population

OBJECTIVENationwide cancer incidence data were used to analyze the trends of cancer incidence in China in order to provide basic information for making cancer control strategy.

METHODSWe retrieved and re-sorted valid cancer incidence data from the National Central Cancer Registry Database over the 20 year-period 1989-2008. Crude incidence rate and age-standardized incidence rate were calculated for analysis. Annual percent changes in incidence for all cancers combined were estimated using Joinpoint software.

RESULTSThe cancer incidence rate in cancer registration areas was increased from 184.81/10(5) in 1989 to 286.69/10(5) in 2008 (from 209.33/10(5) to 307.04/10(5) in urban and from 176.10/10(5) to 269.57/10(5) in rural areas). Uptrends of crude cancer incidence were shown in both male and female in urban and rural areas over the 20 year-period. After standardized by age, overall incidence rate kept stable with 0.5% annual increase in urban and no change in rural areas. Since 2000, the cancer incidences in both sexes and areas were significantly increased. The incidence increased for most major cancers, especially lung cancer, colorectal cancer, female breast cancer and cervical cancer.

CONCLUSIONSOver the 20 year-period 1989-2008, cancer incidence of most cancers has been increasing by time. The incidences of gastric cancer, liver cancer and esophageal cancer still keep gradually increasing. The incidences of lung cancer, female breast cancer, colorectal cancer and cervical cancer are markedly going up, so that cancer prevention and control should be enhanced. Cancer registration will play an important role on cancer control in China along with the number of registries increasing and data quality improving.

Breast Neoplasms ; epidemiology ; China ; epidemiology ; Colorectal Neoplasms ; epidemiology ; Esophageal Neoplasms ; epidemiology ; Female ; Humans ; Incidence ; Liver Neoplasms ; epidemiology ; Lung Neoplasms ; epidemiology ; Male ; Neoplasms ; epidemiology ; Registries ; Rural Population ; Stomach Neoplasms ; epidemiology ; Urban Population ; Uterine Cervical Neoplasms ; epidemiology

To study immunophenotype and cytotoxicity of the immunocytes in bone marrow and peripheral blood after activation by combined cytokines, mononuclear cells (MNC) of bone marrow and peripheral blood were activated by IFN-gamma, IL-1, IL-2 and McAb-CD3 in vitro. The cell amount and morphology during culture were observed. Cytochemical staining and immunophenotype analysis were done before and after culture in two groups of MNC. Cytotoxicity was tested by MTT method. The results showed that the cell number of two groups increased obviously in culture (P < 0.05), while the peripheral blood mononuclear cells increased more markedly (P < 0.05). The cytochemical staining showed POX decrease, but PAS increase in two groups. The positive ratios of CD3(+), CD56(+) and CD38(+) cells in two groups increased obviously after culture (P < 0.05), but there was no significant difference between those two groups. CD3(+) CD56(+) cells increased obviously in peripheral blood mononuclear cells activated by cytokines (P < 0.05), but CD3(+) CD56(+) cells did not increase in bone marrow mononuclear cells. There was no significant difference between two groups' cytotoxicity. It was concluded that IFN-gamma, IL-1, IL-2 and McAb-C D3 increased cell number and cytotoxicity of both bone marrow and peripheral blood mononuclear cells that can be used in cell immunotherapy.
ADP-ribosyl Cyclase ; immunology ; ADP-ribosyl Cyclase 1 ; Antibodies, Monoclonal ; pharmacology ; Antigens, CD ; immunology ; Bone Marrow Cells ; cytology ; drug effects ; immunology ; CD3 Complex ; immunology ; CD56 Antigen ; immunology ; Cell Count ; Cell Division ; drug effects ; Coculture Techniques ; Cytokines ; pharmacology ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; drug effects ; HL-60 Cells ; Humans ; Immunophenotyping ; Interferon-gamma ; pharmacology ; Interleukin-1 ; pharmacology ; Interleukin-2 ; pharmacology ; K562 Cells ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Membrane Glycoproteins ; Time Factors

Objective To investigate the prevalence and characteristics of visual hallucination among patients with Parkinson's disease (PD), and analyse the potential risk factors. Methods One hundred and twenty-eight patients with PD were administered self-prepared visual hallucination questionnaires, and prevalence of visual hallucination was surveyed. The differences in sex, age, disease duration, Mini Mental State Exam (MMSE) scores, Hoehn & Yahr stage, types of medicine used and levodopa equivalent doses (LDE) were compared between the patients with visual hallucination and those without visual hallucination. The prevalence of rapid eye movement sleep behavior disorder (RBD) was investigated in patients with visual hallucination. Results Eighteen patients (14.06%) experienced visual hallucination, among whom 10 (55.56%) experienced visual hallucination no less than one time per day, 11 (61.11%) "saw" the shadow of human figure and 15(83.33%)were complicated with RBD. There were significant differences in MMSE scores, Hoehn & Yahr stage, female proportion and usage of dopamine agonists between patients with visual hallucination and those without visual hallucination(P<0.05). Conclusion Visual hallucination is a common non-motor symptom among patients with PD, and cognitive function, disease severity and usage of dopamine agonista may be related to visual hallucination.

OBJECTIVE: To investigate the effectiveness and safety of laparoscopic CY Liu non-mesh pelvic floor repair surgery in treatment of pelvic organ prolapse(POP).METHODS: From February 2016 to September 2017,a total of 83 patients with POP were treated in Zhejiang Province People's Hospital,and they were included in this retrospective study.The clinical data were retrospectively analyzed;the POP-Q scores were used as the objective evaluation indicators for POP.Analyze the changes of POP-Q indicator points after surgery,and observe surgery related data and complications.Evaluate the postoperative symptom improvement and subjective satisfaction rate of the patients using pelvic floor impact questionnaire short form(PFIQ-7).RESULTS: All surgeries of the included 83 patients were successfully finished laparoscopically.The average follow-up time was(18.88±3.82)months,but there were 8 cases of loss of follow-up.During the follow-up,the total anatomical success rate of laparoscopic CY Liu non-mesh pelvic floor repair surgery was 97.33%(73/75).The differences in POP-Q scores and PFIQ-7 scores were statistically significant before and after the surgery(P<0.05).Subjective satisfaction rate was 96.00%(72/75)after surgery.CONCLUSION: Laparoscopic CY Liu non-mesh pelvic floor repair surgery is effective in the management of Ⅱ-Ⅳ pelvic organ prolapse.There's no need for mesh,and the recurrence rate is low in short-term follow-up,which is worth paying attention to.

OBJECTIVETo determine the relationship between polymorphisms in the genes encoding IL-1, IL-6, and IL-10 with primary biliary cirrhosis (PBC) in Chinese population.

METHODSWhole-blood samples were taken from 77 patients with PBC and 160 healthy controls. DNA was extracted and the polymorphisms at positions IL-1 +3953, IL-1RN intron 2, IL-6 -174, and IL-10 -1082, -819, and -592 were determined by using sequence-specific polymerase chain reaction (SSP) or polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

RESULTSThe frequency of IL-1RN1,1 allele in PBC group was significantly higher than in control group (90.9% vs 79.4%, P=0.026), and the frequency of IL-1RN1,2 in PBC group was significantly lower than in control group (6.5% vs 18.8%, P=0.013). There was no significant difference in the frequence of IL-1RN*2 allele between PBC group and control group (P=0.06). Of the 77 patients with PBC, 4 patients were IL-6 -174GC, 73 were IL-6 174GG. All the 160 health controls are IL-6 -174GG (P=0.0036). The frequence of IL-6 -174C allele in PBC group was significantly higher than that in control group (P=0.0038). No significant differences of polymorphisms for IL-1 +3953 and IL-10 (-1082, -819 and -592) were found between PBC group and control group.

CONCLUSIONThe polymorphisms of IL-1RN and IL-6 -174G/C appear to be associated with PBC, and the polymorphisms of IL-1 +3953 and IL-10 promoter gene are not associated with PBC in a Chinese population.

Adult ; Aged ; Female ; Humans ; Interleukin-1 ; genetics ; Interleukin-10 ; genetics ; Interleukin-6 ; genetics ; Liver Cirrhosis, Biliary ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length

Conditions of disuse such as bed rest, space flight, and immobilization result in decreased mechanical loading of bone, which is associated with reduced bone mineral density and increased fracture risk. Mechanisms involved in this process are not well understood except the suppression of osteoblast function. To investigate the effect of simulated weightlessness on mRNA level of extracellular matrix proteins, osteoblasts were rotated in horizontal plane as a model of simulated microgravity. Primordial osteoblasts of rats were grown for 2 d and then rotated for 24, 48 and 72 h, respectively. After isolating total RNA in cells, reverse transcription polymerase chain reaction (PT-PCR) was made to examine the mRNA level of osteopontin (OPN) and osteonectin (ON). Meanwhile, the levels of alkaline phosphatase (ALP) and osteocalcin (BGP) in the cultured medium were measured to evaluate the calcific function of cell. The expression of OPN and ON mRNA fell significantly after rotating for 24, 48 and 72 h, respectively. The contents of BGP descended significantly, meanwhile, the activity of ALP also showed a degressive tendency. Horizontal rotation decreased the expression of ON and OPN as well as diminished the secretion of BGP and ALP, which affected the calcific function of osteoblast. The results obtained suggest that depression of extracellular matrix proteins expression plays a key role in bone loss during weightlessness.
Animals ; Animals, Newborn ; Cells, Cultured ; Extracellular Matrix Proteins ; biosynthesis ; genetics ; Osteoblasts ; cytology ; metabolism ; Osteonectin ; biosynthesis ; genetics ; Osteopontin ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Rotation ; Sialoglycoproteins ; biosynthesis ; genetics ; Skull ; cytology ; Weightlessness Simulation