AIM: To express chimeric toxin Stx2a’-LHRH a nd to investigate the cytotoxic activity of recombinant toxin Stx2a’-LHRH to huma n carcinoma cells.METHODS: Stx2a’-LHRH sequences that added the res tri ction endonucleases NcoⅠ and EcoRⅠ at the 5' and 3 ends were amplified by PCR a nd digested with appropriate restriction enzymes. The digested fragment was subc loned into the vector obtatined by digestion of plasmid pET-28a(+) with NcoⅠ an d EcoRⅠ. E. coli BL21 (DE3) cells were transformed with plasmids of interst and cultured in LB medium containing ampicillin. Expression of the recombinant protein was induced by the addition of isopropylthio-?-D-galactoside (IPTG). T h e cytotoxity of Stx2a’-LHRH to Hep-2 cells was observed under the microscop y. RESULTS: Recombitant plasmid pET-SL was constructed successfu lly and the clones expressing pET-SL stablely were obtained. A special electroph oretic band in SDS-PAGE (a glycoprotein of 28kD) was noted. Stx2a’-LHRH killed He lp-2 cells clearly. CONCLUSION: In this study, construction of c himeric toxin Stx2a’-LHRH and its expression were described. Moreover, it has o bvious cytotoxity to Hep-2 cell. These finding could open up new vistas in the s tudy of targeted durgs.

OBJECTIVETo explore and compare the clinical effects of three immuno-potentiators and their influence on immune function in preventing and treating recurrent respiratory tract infection (RRI).

METHODSSeventy-two children with RRT were assigned to three groups, the 23 patients in the M group treated with milkvetch extract oral liquid, the 23 in the P group treated by P-transfer factor and the 26 in the U group treated by Utilins injection. Clinical effect was compared among three groups after treatment, and changes of T cell subgroups as well as immune antibodies were detected before and after treatment.

RESULTSThe markedly effective rate in the M, P, and U group was 69.6%, 65.2%, and 73.0% respectively, and the total effective rate 87.0%, 82.6%, and 92.3%, showing an insignificant difference among them (all P>0.05). After treatment, the levels of CD3+, CD4+ and CD4+/CD8+ rose obviously in the three groups (all P<0.05); levels of CD8+ in the M group and the P group lowered significantly (P<0.05, P<0.01), but with no significant difference among three groups in the changes of T cell subgroups (all P>0.05). Serum levels of IgG in all the three groups, IgA in the P group and the M group, and IgM in the M group and the U group rose significantly (P<0.05 or P<0.01), with statistical difference presented in comparing of IgA level in the P group with that in the other two groups (P<0.01).

CONCLUSIONThe clinical effects and the outcomes of immunological indexes' changing are similar in RRI patients treated by the three immuno-potentiators. They all can enhance the immunological function of the organism, strengthen the disease-resistant ability of patients, and reduce the incidence of RRI. Among them, the milkvetch extract oral liquid is worthy of spreading due to its lower cost, simple administration, exemption from painful injection, and good compliance.

Administration, Oral ; Astragalus Plant ; chemistry ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Female ; Humans ; Immunoglobulins ; blood ; Infant ; Male ; Respiratory Tract Infections ; blood ; drug therapy ; immunology ; prevention & control ; Retrospective Studies ; Secondary Prevention ; T-Lymphocyte Subsets ; drug effects ; immunology ; Treatment Outcome

Objective To explore feasibility of the Chinese version of MoCA for the detection of vascular cognitive impairment-no dementia (VaCIND) and control in a cross-sectional study. Methods One hundred and three Chinese Han were assessed by the MoCA and MMSE. 64 met criteria for VaCIND and 39 were considered cognitively normal. Sensitivities and specificities were calculated using the recommended cut-off scores,and ROC curve analyses were performed to determine optimal sensitivity and specificity. Results No differences were found between groups on age,gender,education degrees. According to their MoCA scores,cognitive impairments including memory,visuospatial, executive function, attention, language, and orientation sub-scores in VaCIND ((0.44 ± 0.96), (2.13 ±1.40), (1.90 ±1.02), (4.61 ±1.41), (4.23 ±1.40), (5.38 ±1.15)) significantly decreased compared with that in controls((2.92 ± 1.42) ,(3.16 ± 1.08) ,(3.32 ± 1.07) ,(5. 87 ±0.41) ,(5.34 ±0.75), (5.79 ±0. 70)) (P<0. 05). The MMSE scale was insensitive to cognitive impairment as compared with MoCA scale. Using cut-off score of 24,the MoCA exhibited excellent sensitivity (0.923) and specificity (0.906). Conclusion MoCA is a more sensitive instrument than the MMSE for the detection of VaCIND and warrants further investigation regarding its applicability in large group and varying ethnic groups.

Objective To investigate outcome and cognitive changes of amnestic mild cognitive impairment (aMCI) in a follow-up study. Methods A cross-sectional and longitudinal parallel cohort study design was conducted among 109 aMCI patients and 104 matched normal controls. Multi-dimension neuropsychologic tests were used to extensively assess the cognitive function. Results The scores of neuropsychologic tests in aMCI patients were significantly lower than those in the normal controls( all P＜0.01 ) ,with the largest impairment on 20minutes delayed recall of the auditory verbal memory test ( AVMT), which reflects episodic memory ( aMCI pa-tients :2.50 ± 1.48, normal controls :7.85 ± 1.59, Z = - 12.697, P ＜ 0.01 ); AD was diagnosed in 15 of the 69aMCI patients with a prevalence rate of 22% ,but none was converted to AD in the normal controls. The cognitivechanges of performance in AVMT, CDT, MMSE of the patients in aMCI group (( 3.77 ± 60.83 )%, (6.89 ±28.24) %, (6.13 ± 16.89) % respectively) were significantly poorer than those of the controls group(( - 10.75 ±27.46) %, ( - 5.23 ± 14.05 ) %, ( - 1.11 ± 8.26 ) % respectively) ( all P ＜ 0.05 ). At baseline, demented aMCIperformed poorer in AVMT, CFT, TMT, SDMT, CDT, MMSE when compared to stable. During the follow-up, demented aMCI groups performed significantly poorer than did stable subjects in AVMT, CFT, DST, VFT, SDMT,MMSE ( all P ＜ 0.05 ). Conclusion aMCI is a prodromal period of AD and characterized by episodic memory impairment. The neuropsychologic test is a predictive factor for aMCI to develop AD.

Objective: To estimate the incidence of metabolic syndrome and explore possible risk factors for metabolic syndrome in adults of rural communities in Yuhuan county, Zhejiang province, China. Methods: During June-December, 2018, a follow-up survey was conducted in participants without metabolic syndrome at baseline survey in 2012 to obtain the information collected in questionnaire survey, anthropometric data and laboratory data. The incidence of metabolic syndrome in the participants was estimated, and Logistic regression model was used to explore the risk factors, adjusted risk ratio (aRR) and 95%CI. Results: Among 3 162 participants, 522 new metabolic syndrome cases were identified. The 6-year cumulative incidence rate of metabolic syndrome was 16.5%, and the cumulative incidence rate was higher in women (20.6%) than that in men (12.3%, P<0.001). Those incidence rates were higher in those in jobless, smoking or drinking groups. Being women (aRR=1.96, 95%CI: 1.50-2.58) and family history of hypertension (aRR=1.31, 95%CI: 1.04-1.63) were independent risk factors for metabolic syndrome. Conclusion: The follow up indicated that the incidence of metabolic syndrome was relatively high in rural adults on islands in Zhejiang, and women or those with family history of hypertension were more likely to have metabolic syndrome.
Adult ; Female ; Humans ; Incidence ; Islands ; Male ; Metabolic Syndrome/epidemiology* ; Risk Factors ; Rural Population

OBJECTIVE: One of the objective of the study was to evaluate the reliability and validity of 18 items survey (OSA-18) which was made for disease specific quality of life in children with obstructive sleep apnea. The other was to explore whether OSA-18 could be used as a well appreciation method on OSAHS in children's quality of life before and after surgery. METHOD: First, one hundred and twenty-two children's parents were interviewed with this survey scales and the survey scales was assessed by split-half reliability, retest reliability, internal consistency, construct validity and content validity. Second,OSA-18 was used to evaluate the QOL of 122 children within 4 weeks before PSG and in 6 to 12 months after operation. RESULT: OSA-18 has a satisfactory internal consistency. Global Cronbach's alpha coefficient were 0.939. All domains and items Cronbach's alpha coefficients > 0.6; also has a well retest reliability, coefficient correlation of pearson equal 0. 619. Construct validity and content validity were satisfactory. The impact of pediatric OSAHS on QOL was moderate and severe in 77.05% patients pre-operation. However, it was only 28.69% after surgery. OSA-18 total scores, every domain and item score were significant decrease. CONCLUSION OSA-18 QOL survey had better reliability and validity, and was applicable for life quality assessment in OSAHS children. OSA-18 QOL survey could help clinical diagnosis of OSAHS in children and give quantitative evaluation for therapeutic measure.
Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Parents ; Quality of Life ; Reproducibility of Results ; Sleep Apnea, Obstructive ; Surveys and Questionnaires

Objective To study on the relationship between the serum level of highsensitivity c-reaction protein (hs-CR),matrix metalloproteinase 9 (MMT-9),tissue inhibitor of metalloproteinase 1 (TIMP-1) and carotid plaque stability in elderly people.Methods According to the carotid ultrasonography examination,120 old people were divided into vulnerable plaque group (group A,n =45),stable plaque group (group B,n =41) and no plaque group (group C,n =34),and serum levels of hs-CRP,MMP-9,TIMP-1 in each group were also detected.Results The serum levels of hs-CRP and MMP-9 of unstable plaque group and stable plaque group were higher than those of non-plaque group (P ＜ 0.05).hsCRP and MMP-9 were positively correlated with carotid plaque unstability,while TIMP-1 was negatively correlated with carotid plaque unstability,hs-CRP,MMP-9 and TIMP-1 were independent risk factors for carotid plaque stability.Conclusions Serum levels of hs-CRP,MMP-9 and TIMP-1 are closely related to the stability of carotid plaque.Elevated levels of hs-CRP and MMP-9 increase the risk of carotid plaque,and elevated levels of TIMP-1 decrease the risk of carotid plaque.

To screen the specific anti-human intercellular adhesion molecule-1 (ICAM-1) single chain fragment variable (scFv) using phage display library technology and to identify its biological activity. P1 peptide was used as antigen, and the phage antibodies against human ICAM-1 antigen were panned by four binding-eluting-amplifying cycles using Tomlinson I+J phage display library. After four rounds of selective enrichment screening, the positive clones were determined by PCR, enzyme linked immunosorbent assay (ELISA)-based antigenic cross reaction and Dot blotting. Then the binding specificity and biological activity of purified scFv were identified by Western blotting, competitive ELISA and cell adhesion inhibition assay respectively. Furthermore, four positive clones were first panned through P1 peptide coated-ELISA assay, and then J-A1 was obtained and identified by PCR, ELISA-based antigenic cross reaction and Dot blotting, which could show a specific binding between P1 peptide and human ICAM-1 protein antigen. Subsequently, the purified scFv showed a satisfactory specificity and anti-adhesive activity in competitive ELISA and the cell adhesion inhibition assay. The specific anti-human ICAM-1 scFv was prepared successfully from Tomlinson I+J phage display library, which pave the way for further application of anti-human ICAM-1 scFv for inflammation diseases therapeutics.
Antibodies ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin Variable Region ; Intercellular Adhesion Molecule-1 ; immunology ; Peptide Library ; Single-Chain Antibodies