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 :The study discussed the relationship between the cell differentiation and its func-tion differentiation of enamel organ durrng the development of enamel ,explained the meaning of shape andfunction of SA and RA in the mature enameloblasts. Methods:The rats were fixed with pouring hearts ,em-bed with Epon and GMA resin,made semi-thin slice ,stained with toluidine blue and histochemistry, thenwe observed the changes of histomorphology and the law of distribution of enamel. Results:The develop-memt of tooth germ of rat incisor could be divided into proliferation stage, difference stage and maturestage. Enameloblasts expressed special periodic changes in mature stage :SA legion and RA legion present-ed altermately. Histochemical study indicated that most of amelogenin stayed in enamel matrix and othersdiffused to dentin, dental tube and odontoblast layer. Conclusion: (1) The proliferation stage, differencestage and mature stage of tooth of development of enamel organ in rat were similar to the bud stage ,capstage and bell stage of human being. There was a relationship between the RA legion in mature enam-eloblast and mineral substance pouring into them. RA legion was presented because water and protein lost.)That amelogenin spread into dentin could promote the odontoblast differentiation and induce dentin calci-fication.

Objective To investigate the impact of aging and vascular endothelial function on arterial stiff- ness in patients with isolated systolic hypertension.Methods Patients with isolated systolic hypertension (ISH,n=75)age-matched healthy subjects(n=30)and young healthy subjects(n=50)were submitted to deter- mination of aortic pulse wave velocity(baPWV)and vascular endothelial function evaluated by flow-mediated dila- tion(FMD).Results baPWV was progresively decreased(ISH:2459.2?436.8 vs elderly healthy:2097.2? 315.7 vs young healthy:1619.7?214.2 cm/s,P

Objective:To investigate the predictive value of serum soluble urokinase-type plasminogen activator receptor (suPAR) combined with alpha-fetoprotein (AFP) and model for end-stage liver disease (MELD) score in short-term prognosis assessment of patients with chronic hepatitis B (CHB) related acute-on-chronic liver failure (ACLF).Methods:From January 2018 to May 2020, 66 patients with CHB related ACLF from Fuzhou First People′s Hospital were enrolled. After 90 days of follow-up, the patients with CHB related ACLF were divided into death group and survival group according to the outcome. Meanwhile, 30 patients with CHB were enrolled by simple random sampling method. The differences of serum suPAR in patients with CHB related ACLF and patients with CHB were analyzed. The values of suPAR, AFP and MELD score were compared between death group and survival group in patients with CHB related ACLF. The predictive value of suPAR, AFP, MELD score, Child-Turcotte Pugh score (CTP score) and suPAR combined with AFP and MELD score in the short-term prognosis of patients with CHB related ACLF were analyzed by area under the receiver operator characteristic curve (AUROC). Data were analyzed by two independent sample t test or non-parametric test. Results:The serum suPAR level of patients with CHB related ACLF was (9.6±0.8) ln ng/L, which was higher than that of patients with CHB ((8.0±0.3) ln ng/L). The difference was statistically significant ( t=14.533, P<0.01). The suPAR and MELD score of patients with CHB related ACLF in the death group were (9.9±0.7) ln ng/L and 29.6 (7.1) points, respectively, which were higher than those in the survival group ((9.4±0.7) ln ng/L and 21.0 (5.0) points, respectively). The AFP level in the death group was 45.9 (108.1) μg/L, which was lower than that in the survival group (209.3 (187.1) μg/L). There were significant differences in suPAR ( t=2.895, P=0.005), MELD score ( Z=4.708, P<0.01) and AFP ( Z=3.051, P<0.01) between the death group and the survival group. AUROC of suPAR (0.741, P=0.001), AFP (0.724, P=0.002) and MELD score (0.885, P<0.01) had predictive value for death in patients with CHB related ACLF. The sensitivities of suPAR, AFP, MELD score, CTP score and suPAR combined with AFP and MELD score were 84.6%, 73.1%, 88.5%, 96.2% and 84.6%, respectively, and the specificities were 75.0%, 72.5%, 70.0%, 52.5% and 92.5%, respectively. The AUROC of suPAR combined with AFP and MELD score was 0.871 ( P<0.01), which was higher than that of CTP score (0.793, P<0.01). Conclusions:Serum suPAR is increased in patients with CHB related ACLF. SuPAR combined with AFP and MELD score could apply in the prognostic value for patients with CHB related ACLF.

ObjectiveTo explore the reliability and validity of WHO ICF-Clinical Checklist for patients with spinal cord injuries.MethodsClinical testing, case records and check had been implemented for 50 patients with spinal cord injuries with ICF-Clinical Checklist, ADL and ASIA.Results and ConclusionThere were high test-retest reliability and validity in WHO ICF-Checklist for patients with spinal cord injuries.

Adult ; Aged ; Carcinoma, Squamous Cell ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; pathology ; virology ; Colposcopy ; Cytodiagnosis ; DNA Probes, HPV ; DNA, Viral ; isolation & purification ; Female ; Humans ; Middle Aged ; Nucleic Acid Hybridization ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus Infections ; Uterine Cervical Neoplasms ; pathology ; virology ; Vaginal Smears ; Young Adult

Humans ; Infant, Newborn ; Male ; Neuromuscular Nondepolarizing Agents ; therapeutic use ; Tetanus ; drug therapy ; Vecuronium Bromide ; therapeutic use

Objective To observe the therapeutic efficacy of balance cupping plusYang He decoction in treating granulomatous mastitis (GM).Method Eighty GM patients were randomized into a treatment group and a control group according to the last digit of registered number, odd or even, 40 cases each. The control group was treated withYang He decoction; the treatment group was intervened by balance cupping in addition to the treatment given to the control group. They were treated for 6 months in total. The size of lump, swollen size and infection size by the ultrasound B-scanning were observed before and after the treatment, and the clinical efficacies were compared between the two groups.Result The total effective rate was 82.5％ in the treatment group versus 60.0％ in the control group, and the between-group difference was statistically significant (P<0.05). There was a significant difference in comparing the clinical efficacy between the two groups (P<0.05). The size of lump, swollen area and infection size by the ultrasound B-scanning were significantly changed in the two groups after the treatment (P<0.05); the between-group comparisons showed significant differences after the treatment (P<0.05).Conclusion Balance cupping plusYang He decoction can produce a certain efficacy in treating GM.

The growing status of Anoectochilus roxburghii seedling was observed and the survival rate of seedlings, height, stem diameter and plant fresh weight under the conditions of different transplanting substrate compositions, planting density, shading rate were measured. The results showed that the effects of different transplanting substrates, planting densities, shading rates and nutrient solutions on the growing status of A. roxburghii plantlets varied greatly. A. roxburghii plantlets demonstrated a high survival rate and better growing status under the Following conditions: the ratio of peat and river sand as 2: 1, the planting density as 3 cm x 3 cm, the shading rate as 70%, and the nutrient solution as 1/4MS. The findings of the study provide a solid technical solution for the artificial cultivation of A. roxburghii plantlets.
Breeding ; methods ; Culture Media ; chemistry ; pharmacology ; Orchidaceae ; drug effects ; growth & development ; Seedlings ; drug effects ; growth & development ; Survival Analysis

The aim of this study was to investigate the in vitro effect of interleukin-24 (IL-24) on apoptosis of bone marrow mononuclear cells (BMMNC) in children with acute leukemia. Every group of acute lymphocytic leukemia (ALL) and acute myeloid leukemia (ANLL) had 20 children who did not receive any therapy. The bone marrow was taken from patients and controls, the MNC were isolated from bone marrow, DNA was detected by glucose electrophoresis. Apoptosis of BMMNC was assayed by flow cytometry with propidium iodine staining. RT-PCR was used to detect the expression level of bcl-2, caspase-3 mRNA, and to analyze the effect of IL-24 on them. The results showed that the IL-24 induced apoptosis of BMMNC in children with acute leukemia. After acute leukemia BMMNC were exposed to IL-24 for 48 hours, DNA ladder fragment appeared, and the apoptotic rate of the group treated with IL-24 of 50 ng/ml was obviously higher than that of the control group (0 ng/ml). IL-24 decreased the bcl-2 mRNA expression level, enhanced caspase-3 mRNA expression level of BMMNC from AL patients. It is concluded that the IL-24 can induce apoptosis of AL BMMNC in vitro, which may be due to decreasing of bcl-2 mRNA level and enhancing of caspase-3 mRNA level.
Acute Disease ; Adolescent ; Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; drug effects ; Caspase 3 ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Interleukins ; pharmacology ; Leukemia ; pathology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism