Aims To construct the N-terminal Strep-tagged ( NS-tagged) fusion protein expression vector, and to apply the vector to express NS-tagged fusion proteins of Chlamydia RNA polymerase subunit. Meth-ods By using PCR method, NS fusion protein tag and a new multiple cloning sites (MCS) were inserted into pET21c-DH plasmid by primers to replace the original T7 protein tag and MCS. The newly introduced Not I cutting site was chosen for self-ligation of PCR prod-uct. Then, the cyclized PCR product was transformed into DH-5α competent cells. The positive clones were selected by PCR and sequencing. To get NS-tagged fu-sion proteins of chlamydial RNA polymerase subunits, the α, β and β′ subunits were inserted between BamH I and Sal I cutting sites of the newly constructed ex-pression vector. Then, the NS-α, NS-β and NS-β′ ex-pression vectors were transformed into Arctic Express expression cells. The fusion protein expression statuses of transformed cells were identified by Commassie blue staining and Western blot. Results The NS-tagged fusion protein expression vector pET21c-NS-MCS was successfully constructed, and NS-α, NS-β and NS-β′fusion proteins were obtained by using this newly con-structed expression vector. Conclusions In this pro-ject, we constructed an NS-tagged fusion protein ex-pression vector and applied it to express NS-α, NS-βand NS-β′ fusion proteins. Our study can lay a solid foundation for the study of transcriptional regulation of Chlamydia genes.

Allergic diseases such as allergic asthma, allergic der-matitis, allergic rhinitis, are polygenic diseases, involving the interaction between the environment, genes and immunity. In the past few decades, the incidence rate of allergic diseases in-creased predominantly and influenced the quality of people's lives seriously, so looking for new targets for the prevention and treat-ment of allergic diseases and drugs with less adverse reaction be-comes a hot topic for researchers. MicroRNAs(miRNAs)are a class of endogenous non-coding small RNAs that mediate nega-tively posttranscriptional regulation of gene expression by targe-ting specific mRNA sequences to inhibit the translation of mR-NAs. They are widely involved in the biological processes of cell differentiation, immune response and tumor development. The study shows that miRNAs can control the occurrence and devel-opment of allergic diseases. Studying the regulatory role of miR-NAs in allergic diseases has important implications for exploring the immunopathological mechanisms and discovering new thera-peutic targets of drugs.

Aim To investigate the method of establishing the atopic dermatitis mice model induced by calcipotriol ( MC903 ) . Methods Induction dose exploratory experiment: since d 0, 0.33,1,3 nmol MC903 was smeared on the right ear of BALB/c mice in each dose group respectively , for 7 consecutive days . The ear swelling degree of the mice was observed every day and the bilateral ears thicknesses were measured .The materials were drawn and analyzed in d 3 and d 7.Induction days exploratory experiment: 2 nmol MC903 was smeared on the right ear of BALB/c mice, for 14 consecutive days .The ear swelling degree of mice was observed every day and the bilateral ears thicknesses were measured .The histopathological examination of the right ear was conducted in 3 d, 7 d, 11 d and 15 d respectively .The tissue homogenate of the mice right ear was prepared .The ex-pressions of thymic stromal lymphopoietin (TSLP),IL-33, IL-4 and IFN-γin the homogenate , CD40 +,CD86 +,the DC surface markers and ILC2 contents in the peripheral lymph nodes were detected.Results ①1,3 nmol MC903 induced ear swelling in mice was significantly increased in d 3, the levels of TSLP and IL-4 were significantly increased .The level of IL-33 in 3 nmol dose group was increased significantly in d 7.② The right ear swelling of 2 nmol MC903 induced atopic dermatitis mice was significant , the ear thickness was increased gradually and reached the peak in d 14.The histopathological examination of the mice ear tissue showed in d 7, the right ear tissue of the mice was swelling and red , the capillary vessels were dilated and the infiltration of inflammatory cells was obvious .The ear in-flammatory symptoms maintained and gradually aggravated for 15 days.Compared with the normal mice , MC903 increased the TSLP level in the right ear tissue homogenate significantly in d 3 and then decreased gradually .The levels of IL-4 and IL-33 were increased significantly in d 7 and then decreased gradually .The levels of ILC2, CD40 +, CD86 + in the peripheral lymph nodes were increased in d 7 and d 15.Conclusion The atopic derma-titis mice model can be successfully established using 2 nmol MC903 induced mice for 7 days.Appropriate testing point of TSLP is d 3.Appropriate testing point of IL-33 and IL-4 is d 7.

AIM To optimize the combined enzymatic extraction for alkaloids and polysaccharides from Dendrobium nobile Lindl..METHODS With enzyme consumption,enzymolysis temperature,enzymolysis time and solidliquid ratio as influencing factors,contents of dendrobine,total alkaloids and polysaccharides as evaluation indices,orthogonal test was applied to optimizing the combined enzymatic extraction.RESULTS The optimal conditions for papain extraction were determined to be 0.10 g for enzyme consumption,45 ℃ for enzymolysis temperature,2 h for enzymolysis time,and 1 ∶ 50 for solid-liquid ratio,the contents of dendrobine,total alkaloids and polysaccharides were 3.495 5,4.341 8 and 35.898 7 mg/g,respectively.The optimal conditions for cellulase extraction were determined to be 0.30 g for enzyme consumption,50 ℃ for enzymolysis temperature,2 h for enzymolysis time,and 1 ∶40 for solid-liquid ratio,the contents of three constituents were 3.514 8,4.351 3 and 36.331 2 mg/g,respectively.The optimal conditions for pectinase extraction were determined to be 0.45 g for enzyme consumption,55 ℃ for enzymolysis temperature,2.5 h for enzymolysis time,and 1 ∶ 40 for solid-liquid ratio,the contents of three constituents were 3.524 4,4.452 8 and 26.324 2 mg/g,respectively.CONCLUSION This stable and reliable method can be used for the rapid combined enzymatic extraction for alkaloids and polysaccharides from D.nobile.

Objective To evaluate peak left ventricular(LV) apical rotation velocity during early diastole by speckle tracking echocardiography in normal peopleand patients with myocardial hypertrophy because of hypertension. Methods Two dimensional images of left basal and apical ventricular short axis view were recorded in 20 healthy people and 20 patients with hypertension and left ventricular hypertrophy(LVH). Rotation velocity-time curves of six segments in LV apex and base were obtained using Qlab software. The peak rotation velocity in early diastole of LV apex(A-Vel) and base (B-Vel) were acquired through Excel software. Results In LVH group,A-Vel was significantly decreased(P<0. 05) while B-Vel appeared no difference (P = 0.58) compared with healthy individuals. Conclusions Peak LV apical rotation velocity in early diastole can evaluate the change of LV diastolic function in patients with hypertension and LVH.

OBJECTIVETo establish a quantitative method of simultaneously determination of swertiamarin, gentiopicroside, mangiferin, swertianolin, isoorientin, 1,8-drihydroxy-3-methoxy-xthanone in Swertia from Qinghai province and Sichuan province by HPLC.

METHODThe samples were separated on the column of Kromasil C18 (4. 6 mm x 250 mm, 5 microm) which eluted with methanol and water (content 0.02% phosphoric acid). The ratio of methanol increased from 20% to 80% during 20-50 min, and from 80% to 100% during 50-60 min, with detected wavelength 254 nm, flow rate at 1 mL x min(-1), column temperature 35 degrees C.

RESULTSix compounds were base-isolated, the linear ranges of swertiamarin, gentiopicroside, mangiferin, 4-swertianolin, 5-isoorientin, 1,8-drihydroxy-3-methoxy-xthanone were excellent.

CONCLUSIONThe method was rapid and precise, and can be use for controlling medicinal materials quality.

China ; Chromatography, High Pressure Liquid ; methods ; Glucosides ; analysis ; Iridoid Glucosides ; Iridoids ; analysis ; Luteolin ; analysis ; Plants, Medicinal ; chemistry ; Pyrans ; analysis ; Pyrones ; analysis ; Quality Control ; Reproducibility of Results ; Swertia ; chemistry ; Xanthones ; analysis

OBJECTIVETo observe clinical therapeutic effect of scalp-acupuncture combined with exercise therapy on spastic cerebral palsy.

METHODSEighty children of spastic cerebral palsy were randomly divided into a scalp-acupuncture plus exercise therapy group and a exercise therapy group, 40 cases in each group. The scalp-acupuncture plus exercise therapy group were treated with scalp-acupuncture and exercise therapy, with Yundongqu (the motor area), Pinghengqu (the balance area), Ganjuequ (the sensory area), etc. selected for scalp-acupuncture, and puncture at main points Baihui (GV 20) and Sishencong (EX-HN 1) and exercise therapy. The exercise therapy group were treated by exercise therapy. Changes of GMFM scores and WeeFIM scores before and after treatment were compared.

RESULTSThere were significant differences in GMFM scores and WeeFIM scores before and treatment in the scalp-acupuncture plus exercise therapy group (P < 0.001) and in the exercise therapy group (P < 0.05), the former being better than the later (P < 0.05); the total effective rate was 92.5% in the scalp-acupuncture plus exercise therapy group and 72.5% in the exercise therapy group with a significant difference between the two groups (P < 0.05), the former being significantly higher than the later.

CONCLUSIONThe scalp-acupuncture combined with exercise therapy can improve motor function of limits of children with spastic cerebral palsy, with therapeutic effect better than that of simple exercise therapy.

Acupuncture Therapy ; Cerebral Palsy ; physiopathology ; rehabilitation ; therapy ; Child ; Child, Preschool ; Combined Modality Therapy ; Exercise Therapy ; Female ; Humans ; Male ; Meridians ; Motor Activity

Background Hyperuricemia is frequently present in patients with heart failure. Many pathological conditions, such as tissue ischemia, renal function impairment, cardiac function impairment, metabolic syndrome, and inflammatory status, may impact uric acid (UA) metabolism. This study was to assess their potential relations to UA metabolism in heart failure. Methods We retrospectively assessed clinical characteristics, echocardiological, renal, metabolic and inflammatory variables selected on the basis of previous evidence of their involvement in cardiovascular diseases and UA metabolism in a large cohort of randomly selected adults with congestive heart failure (n = 553). By clustering of indices, those variables were explored using factor analysis. Results In factor analysis, serum uric acid (SUA) formed part of a principal cluster of renal functional variables which included serum creatinine (SCr) and blood urea nitrogen (BUN). Univariate correlation coefficients between variables of patients with congestive heart failure showed that the strongest correlations for SUA were with BUN (r = 0.48, P < 0.001) and SCr (r = 0.47, P < 0.001). Conclusions There was an inverse relationship between SUA levels and measures of renal function in patients with congestive heart failure. The strong correlation between SUA and SCr and BUN levels suggests that elevated SUA concentrations reflect an impairment of renal function in heart failure.

OBJECTIVETo investigate the influences of bicortical anchorage on values of natural frequencies of dental implants utilizing the 3-dimensional finite element analysis.

METHODSUsing the commercial code of Solidworks, 3-D models of a screw-shaped dental implant and a mandibular bone segment were generated. After the 3-D implant-bone complex was meshed by ABAQUS software, effects of bicortical anchorage on the buccolingual and axial first-order natural frequencies of the implant were computed.

RESULTSBicortical anchorage increased both the buccolingual and axial natural frequencies remarkably. As the bicortical anchorage got deeper, the frequencies correspondingly got higher.

CONCLUSIONBicortical anchorage can increase the buccolingual and axial primary stability of dental implants.

Dental Implantation, Endosseous ; Dental Implants ; Dental Stress Analysis ; Finite Element Analysis ; Humans ; Mandible

Bronchial asthma is a kind of respiratory disease which affects people 's life quality seriously. Many factors in-volved in the occurrence and development of such disease, of which the aberrant expression of E-cad plays a critical role in it. Research found that E-cad is an important cell adhesion molecu-lar, and its main function is to maintain the structural integrity of cells and participate in the improvement of airway remodeling as well as restoration of immune function. Further study showed that the role of mucosal barrier of airway epithelial cells in bronchial asthma patients was often damaged. Moreover, the protein ex-pression of E-cad decreased significantly in mucosal molecular, which suggested that the abnormal expression of E-cad was in-volved in the development of bronchial asthma. A review on the relations between the abnormal expression of E-cad protein and bronchial asthma has been discussed in this paper, also it in-cludes the discussion about the mechanisms of E-cad’ s disorder-induced bronchial asthma as well as explores the strategies of bronchial asthma treatment, which may provide references for the follow-up research and clinical treatment.