Objective:To identify the epitope of mAb15A11 which is specific against RA associated autoantigen cartilage oligomeric matrix protein ( COMP ).Methods: A filamentous phage library displaying random linear dodecapeptides was used to mapping the epitope of mAb15A11.After three rounds of screenings,40 phage clones were selected at random and sequenced.The specificity of phages was confirmed by enzyme immunoassays.Homology search by ClustalW2 and structure analysis by PyMol to identified the epitope amino acid sequence.Western blot analysis of COMP and ELISA analysis of COMP-derived peptides were used to confirm epitope′s characterization.Results: After repeated screenings using bio-panning method, 2 clones were identified, which interacted specifically with mAb 15A11.Homology search did not find succession consensus sequence within COMP molecular,which indicated that the epitope was not linear.PyMol Structure analysis identified the rationality of conformational epitope.Western blot analysis and ELISA of EDTA-treated COMP further prove an conformational structure of the epitope recognized by mAb 15A11.ELISA analysis of COMP-derived peptides demonstrated both disulfide bonds between 229 C-243 C and 237 C-253 C and every epitope amino acid of 232 G,238 H,240 H,241 A,244 V,247 R and 251 R were essential to the binding of mAb 15A11 with COMP.Conclusion: In this study, the potential B cell antigentic epitopes of mAb 15A11 was identified by phage display library.The epitope amino acids sequence and char-acterization were also recognized.It may have important theoretical value for the study of reaction mechanism of COMP antibody and antigen and may also show application significance in the detection of rheumatoid arthritis.

Objective To study the effects of using 25 kDa linear and branched PEI to transfer plasmid DNA pEGFP -C1 (pDNA ,encoding the enhanced green fluorescent protein reporter gene ) to the basilar membrane of the C57BL/6 mice cochlea in vitro .Methods L -PEI/pDNA and B -PEI/pDNA polyplexes were generated in 0 .1M phosphate buffer solution (PBS) or 5% glucose solution .Polyplexes were characterized by transmission electron mi-croscopy .Agarose gel retardation assay was used to determine the plasmid binding ability of L -PEI and B -PEI . The toxicity was investigated by MTT assay .The transfection was firstly evaluated in 293T cell line ,and then the appropriate amount of PEI and plasmid were applied for cochlear explant transfection of P4 mice pups .Results Un-der the same condition ,B -PEI had better transfection efficiency than L -PEI ,but its toxicity was also higher . When generated in PBS ,the polyplexes had lower toxicity than in glucose solution .L -PEI-pDNA nanoparticles could transfect the spiral limbus fibrocytes ,some spiral ganglion neurons and supporting cells ,but the efficiency was low .Conclusion L -PEI could be used as the non -viral vector for the transfection of the cultured basilar mem-brane of P4 mice pups ,but it should be modified to reach higher efficiency .

OBJECTIVETo develop a brief version scale with good validity and reliability to evaluate self-management knowledge, attitude and behavior (KAB) of diabetes patients in a shorter time.

METHODS20 authority experts who specialized in the field of diabetes clinical and non-communicable disease self-management, and familiar with the relevant content of diabetes self-management were selected. Face to face and Email consultation methods were applied in two rounds delphi expert consultations to choose items from the Diabetes Self-management Knowledge, Attitude, and Behavior Assessment Scale (DSKAB, total scale, 98 items). In the first round of expert consultation, experts were asked to select the core items from every dimension and score the extent of familiarity and basis of judgment. In the second round of expert consultation, the significance of the core items which were selected by experts in first round, were graded. The items which entered into the brief scale were determined by harmonious condition of expert opinions.

RESULTSTwo rounds of consultation reclaimed 16 and 15 responses respectively, Positive coefficients of experts were 80% and 94% respectively, and both authoritative coefficients were 0.90. In the first round of consultation, experts selected 44.3 ± 6.2 items on average, 42 items were selected according to the integrity of the scale for measuring dimension and content at last. In the second round of consultation, experts thought that the 42 items could measure the important contents of each dimension comprehensive and representative, the item importance scores of knowledge, attitude, and behavior subscale were 8.42 ± 1.17, 8.61 ± 1.24, 8.61 ± 1.02 respectively, and the coordination coefficients of the three subscale were 0.36, 0.20, 0.49 (χ(2) were 89.74, 11.13, 96.03, P<0.05). The total coordination coefficient was 0.38 (χ(2) =199.36, P<0.001), which indicated the concentration of experts(')opinion was acceptable, showed the brief scale with good face validity and content validity.

CONCLUSIONThe brief scale could evaluate Diabetes Self-management KAB Scale rapidly and relatively comprehensive in groups or individuals diabetes with good face validity and content validity.

Diabetes Mellitus ; therapy ; Health Knowledge, Attitudes, Practice ; Humans ; Reproducibility of Results ; Self Care ; Surveys and Questionnaires

Objective: To investigate the relationship between the erythrocyte membrane protein gene mutations and the clinical severity of hereditary spherocytosis (HS). Methods: Targeted sequencings were performed on 25 HS patients, correlation between HS mutations and patients’ clinical characteristics were evaluated. Results: A total of 25 HS patients were enrolled, including 13 males and 12 females with median age of 20 (4-55) years, including 9 compensatory hemolysis patients, 9 patients with mild anemia, 3 patients with moderate anemia and 4 patients with severe anemia. Of them, 18 patients (72%) harbored HS-related mutations, including ANK1 mutation in 6 cases, SLC4A1 mutation in 6 cases, SPTB mutation in 5 cases and 1 case with EPB41 mutation. Seven patients (28%) didn’t carry common HS mutations. SPTB and SLC4A1 mutations mainly affected male patients. There was no significant difference between the age of diagnosis (P=0.130) and HGB level (P=0.585) in patients with HS mutation and those without mutation, however, the EMA binding fluorescence intensity (P=0.015), AGLT50 (P=0.032) and EOF minimal hemolytic concentration (P=0.027) were significantly different in these two groups of HS patients. Conclusion To screen erythrocyte membrane protein coding gene mutations could favor the diagnosis of HS, and patients without mutations have mild clinical phenotype.

Objective To evaluate the validity, reliability, and acceptability of the brief version of the self-management knowledge, attitude, and behavior (KAB) assessment scale for diabetes patients. Methods Diabetes patients who were managed at the Xinkaipu Community Health Service Center of Tianxin in Changsha, Hunan Province were selected for survey by cluster sampling. A total of 350 diabetes patients were surveyed using the brief scale to collect data on knowledge, attitudes, and behaviors of self-management. Content validity was evaluated by Pearson correlation coefficient between the brief scale and subscales of knowledge, attitude, and behavior. Structure validity was evaluated by factor analysis, and discrimination validity was evaluated by an independent sample t-test between the high-score and low-score groups. Reliability was tested by internal consistency reliability and split-half reliability. The evaluation indexes of internal consistency reliability were Cronbach's α coefficients, θ coefficient, and Ω coefficient. Acceptability was evaluated by valid response rate and completion time of the brief scale. Results A total of 346(98.9%) valid questionnaires were returned, with average survey time of (11.43±3.4) minutes. Average score of the brief scale was 78.85 ± 11.22; scores of the knowledge, attitude, and behavior subscales were 16.45 ± 4.42, 21.33 ± 2.03, and 41.07 ± 8.34, respectively. Pearson correlation coefficients between the brief scale and the knowledge, attitude, and behavior subscales were 0.92, 0.42, and 0.60, respectively;P-values were all less than 0.01, indicating that the face validity and content validity of the brief scale were achieved to a good level. The common factor cumulative variance contribution rate of the brief scale and three subscales was from 53.66%to 61.75%, which achieved more than 50%of the approved standard. There were 11 common factors;41 of the total 42 items had factor loadings above 0.40 in their relevant common factor, indicating that the brief scale and three subscales had good construct validity. Patients were divided into a high-score group and a low-score group, then scores of the brief scale and three subscales were compared between the groups using a t-test. The results were all significant, indicating that the brief scale and three subscales had good discriminate validity. Mean scores of the brief scale and three subscales of the high-score group were 91.55±6.81, 19.51±2.17, 22.74±1.88, and 49.30±6.20, respectively;these were higher than the low-score group (65.89 ± 5.79, 12.29 ± 4.76, 20.22 ± 1.88, and 33.39 ± 6.17, respectively) with t-values 27.76, 13.31, 9.20, and 17.56 (P-values were less than 0.001). The Cronbach's α coefficient, θ coefficient,Ω coefficient, and split-half reliability of the brief scale were 0.83, 0.87, 0.96, and 0.84, respectively. These values for the three subscales were all above 0.70, except for theθcoefficient of the attitude subscale with 0.64, indicating that the brief scale and three subscales had acceptable internal consistency reliability. Conclusion The brief version of the diabetes self-management knowledge, attitude, and behavior assessment scale showed good acceptability, validity, and reliability, to responsibly evaluate self-management KAB among patients with diabetes.

Objective To evaluate the validity, reliability, and acceptability of the brief version of the self-management knowledge, attitude, and behavior (KAB) assessment scale for diabetes patients. Methods Diabetes patients who were managed at the Xinkaipu Community Health Service Center of Tianxin in Changsha, Hunan Province were selected for survey by cluster sampling. A total of 350 diabetes patients were surveyed using the brief scale to collect data on knowledge, attitudes, and behaviors of self-management. Content validity was evaluated by Pearson correlation coefficient between the brief scale and subscales of knowledge, attitude, and behavior. Structure validity was evaluated by factor analysis, and discrimination validity was evaluated by an independent sample t-test between the high-score and low-score groups. Reliability was tested by internal consistency reliability and split-half reliability. The evaluation indexes of internal consistency reliability were Cronbach's α coefficients, θ coefficient, and Ω coefficient. Acceptability was evaluated by valid response rate and completion time of the brief scale. Results A total of 346(98.9%) valid questionnaires were returned, with average survey time of (11.43±3.4) minutes. Average score of the brief scale was 78.85 ± 11.22; scores of the knowledge, attitude, and behavior subscales were 16.45 ± 4.42, 21.33 ± 2.03, and 41.07 ± 8.34, respectively. Pearson correlation coefficients between the brief scale and the knowledge, attitude, and behavior subscales were 0.92, 0.42, and 0.60, respectively;P-values were all less than 0.01, indicating that the face validity and content validity of the brief scale were achieved to a good level. The common factor cumulative variance contribution rate of the brief scale and three subscales was from 53.66%to 61.75%, which achieved more than 50%of the approved standard. There were 11 common factors;41 of the total 42 items had factor loadings above 0.40 in their relevant common factor, indicating that the brief scale and three subscales had good construct validity. Patients were divided into a high-score group and a low-score group, then scores of the brief scale and three subscales were compared between the groups using a t-test. The results were all significant, indicating that the brief scale and three subscales had good discriminate validity. Mean scores of the brief scale and three subscales of the high-score group were 91.55±6.81, 19.51±2.17, 22.74±1.88, and 49.30±6.20, respectively;these were higher than the low-score group (65.89 ± 5.79, 12.29 ± 4.76, 20.22 ± 1.88, and 33.39 ± 6.17, respectively) with t-values 27.76, 13.31, 9.20, and 17.56 (P-values were less than 0.001). The Cronbach's α coefficient, θ coefficient,Ω coefficient, and split-half reliability of the brief scale were 0.83, 0.87, 0.96, and 0.84, respectively. These values for the three subscales were all above 0.70, except for theθcoefficient of the attitude subscale with 0.64, indicating that the brief scale and three subscales had acceptable internal consistency reliability. Conclusion The brief version of the diabetes self-management knowledge, attitude, and behavior assessment scale showed good acceptability, validity, and reliability, to responsibly evaluate self-management KAB among patients with diabetes.

Objective:To examine associations of sex hormone levels with bone turnover markers(BTMs) among men in the Northeast region of Henan Province.Methods:From December 2015 to March 2016, 707 male subjects were selected from a National Epidemiological Survey-2014(Thyroid Disorders, Iodine status and Diabetes, TIDE)research—Henan sub-center survey by using multistage stratified cluster random sampling. Fasting venous blood was collected to determine the levels of luteinizing hormone(LH), follicle stimulating hormone(FSH), estradiol(E 2), testosterone(T), dehydroepiandrosterone-sulfate (DHEAS), androstenedione(AD), sex hormone binding globulin(SHBG), dihydrotestosterone(DHT), free testosterone(FT), osteocalcin(OC), pro-collagen type 1 N-terminal propeptide(PINP), C-terminal-cross-linking telopeptide of type 1 collagen(β-CTX), 25-hydroxyvitamin D [25(OH)D], and parathyroid hormone(PTH). Results:A total of 697 men with an average age of(46.6±15.9)years were included in the study. Pearson correlation analysis showed that age was positively associated with LH, FSH, T, and SHBG, while negatively associated with E 2, DHEAS, AD, FT, β-CTX, OC, and PINP, without significant correlation with DHT, 25(OH)D, and PTH. Pearson correlation analysis and linear regression analysis showed that E 2 was negatively associated with β-CTX; T was positively associated with OC, FSH was negatively associated with OC; LH, FSH, and SHBG were negatively associated with PINP; E 2, T, FT, DHT, and AD were positively associated with PINP. After adjusting for age and BMI, linear regression analysis showed that T was still significantly positively associated with OC and PINP, with 0.302 ng/ml and 0.015 ng/ml increasing for OC and PINP every 1 ng/ml increase in T; E 2 and DHT were positively associated with PINP, with 0.250 and 0.047 ng/ml increasing for PINP every 1 pg/ml increase in E 2 and DHT. Conclusions:Age is an important factor influencing sex hormones and BTMs. Serum levels of T, E 2, and DHT are associated with bone formation and bone absorption markers.

OBJECTIVETo report two cases of chronic disseminated intravascular coagulation associated (DIC) with aortic dissecting aneurysm, and discuss the treatment strategy.

METHODSThe clinical data of two patients with chronic DIC associated with aortic dissecting aneurysm were analyzed and the related literature was reviewed.

RESULTSCase 1: female, 53 years old, she had gingival bleeding, skin ecchymosis and haematuria for 2 months, the laboratory test revealed PLT 48 × 10⁹/L, APTT 38.0 s and fibrinogen 0.53 g/L; Case 2: male 86 years old, he had skin petechia and ecchymosis,gingival bleeding for 2 weeks, the laboratory test revealed PLT 17×10⁹/L, APTT 37.5 s and fibrinogen 0.51 g/L. CT scan for both cases revealed aortic aneurysm. They were diagnosed as aortic aneurysm associated chronic DIC. Both of them received blood component transfusion. After the treatment, they showed improvement in bleeding symptoms and laboratory data. They gave up operation, and were discharged from the hospital at last.

CONCLUSIONBlood replacement can alleviate bleeding tendency in those patients with chronic DIC associated with aortic dissecting aneurysm.

Aged, 80 and over ; Aneurysm, Dissecting ; complications ; Aortic Aneurysm ; complications ; Blood Coagulation Disorders ; etiology ; Female ; Hemorrhage ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed

Background: Subclinical hypothyroidism (SCH) is the most common thyroid dysfunction, and its relationship with blood pressure (BP) has been controversial. The aim of the study was to analyze the association between SCH and newly-diagnosed hypertension. Methods: Based on data from the Thyroid disease, Iodine nutrition and Diabetes Epidemiology (TIDE) study, 49,433 euthyroid individuals and 7,719 SCH patients aged ≥18 years were enrolled. Patients with a history of hypertension or thyroid disease were excluded. SCH was determined by manufacturer reference range. Overall hypertension and stage 1 and 2 hypertension were diagnosed according to the guidelines issued by the American College of Cardiology/American Heart Association in 2017. Results: The prevalence of overall hypertension (48.7%), including stage 1 (28.9%) and 2 (19.8%) hypertension, increased significantly in SCH patients compared with euthyroid subjects. With elevated serum thyroid stimulating hormone (TSH) level, the hypertension prevalence also increased significantly from the euthyroid to different SCH subgroups, which was more profound in females or subjects aged <65 years. The age- and sex-specific regression analysis further demonstrated the same trends in the general population and in the 1:1 propensity matched population. Similarly, several BP components (i.e., systolic, diastolic, and mean arterial BP) were positively associated with TSH elevation, and regression analysis also confirmed that all BP components were closely related with SCH in female subjects aged <65 years. Conclusion The prevalence of hypertension increases for patients with SCH. SCH tends to be associated with hypertension and BP components in females younger than 65 years.

Background: Subclinical hypothyroidism (SCH) is the most common thyroid dysfunction, and its relationship with blood pressure (BP) has been controversial. The aim of the study was to analyze the association between SCH and newly-diagnosed hypertension. Methods: Based on data from the Thyroid disease, Iodine nutrition and Diabetes Epidemiology (TIDE) study, 49,433 euthyroid individuals and 7,719 SCH patients aged ≥18 years were enrolled. Patients with a history of hypertension or thyroid disease were excluded. SCH was determined by manufacturer reference range. Overall hypertension and stage 1 and 2 hypertension were diagnosed according to the guidelines issued by the American College of Cardiology/American Heart Association in 2017. Results: The prevalence of overall hypertension (48.7%), including stage 1 (28.9%) and 2 (19.8%) hypertension, increased significantly in SCH patients compared with euthyroid subjects. With elevated serum thyroid stimulating hormone (TSH) level, the hypertension prevalence also increased significantly from the euthyroid to different SCH subgroups, which was more profound in females or subjects aged <65 years. The age- and sex-specific regression analysis further demonstrated the same trends in the general population and in the 1:1 propensity matched population. Similarly, several BP components (i.e., systolic, diastolic, and mean arterial BP) were positively associated with TSH elevation, and regression analysis also confirmed that all BP components were closely related with SCH in female subjects aged <65 years. Conclusion The prevalence of hypertension increases for patients with SCH. SCH tends to be associated with hypertension and BP components in females younger than 65 years.