Objective To detect the antibodies against recombinant chlamydial plasmid-encoded protein 3(rPgp3),chlamydial protease-like activity factor(rCPAF),Ct143 encoded protein(rCT143), Ct101 encoded protein(rCT101),Ct694 encoded protein(rCT694),Ct813 encoded protein(rCT813), Chlamydia membrane protein A(rIncA),Ct875 encoded protein(rCT875),major outer membrane protein (rMOMP)and heat shock protein 60( rHsp60)in serum samples collected form patients with urogenital Chlamydia trachomatis(Ct)infection and to evaluate the antigenicity of those proteins. Methods The re-combinant plasmids expressing the 10 proteins and a blank plasmid were transformed into E. coli BL21 strains,respectively. The transformed E. coli BL21 strains were induced by isopropyl β-D-1-thiogalactopyr-anoside(IPTG)to express recombinant proteins. The glutathione pre-coated 96-well ELISA plates were coa-ted with lysates. Serum samples were collected from 50 patients with Ct infection and 10 patients without Ct infection. ELISA was performed to detect the antibodies against 10 recombinant proteins. Results The anti-bodies against rPgp3,rCPAF,rCT143,rCT101,rCT694,rCT875,rCT813,rMOMP,rIncA and rHsp60 proteins were respectively detected in 44 cases(88% ),38 cases(76% ),37 cases(74% ),36 cases (72% ),33 cases(66% ),31 cases(62% ),30 cases(60% ),26 cases(52% ),24 cases(48% )and 17 cases(34% )out of 50 serum samples. No antibodies against 10 recombinant proteins were detected in the serum samples collected from patients without Ct infection. Conclusion The rPgp3 protein showed the strongest antigenicity among all of the studied proteins,followed by rCPAF and rCT143 proteins. The rHsp60 protein showed the lowest antigenicity.

Objective To optimize immunodominant protein combinations for serological screening for Cblamydia trachomatis (Ct) infection.Methods Both serum and genital swab samples were collected from 50 patients with Ct infection confirmed by colloidal gold immunochromatographic assay (GICA),and 30 GICA-negative clients without Ct infection at a sexually transmitted disease (STD) clinic in Tianjin Medical University General Hospital.The 30 serum samples from GICA-negative clients were also negative for microimmunofluorescence (MIF) assay.Eight Ct immunodominant proteins,including Pgp3,CPAF,CT143,CT101,CT694,CT875,CT813 and IncA,were selected as antigens to detect corresponding antibodies in the serum samples by enzyme-linked immunosorbent assay (ELISA) with the Ct proteins Hsp60 and major outer membrane protein (MOMP) as references.The results of ELISA were compared with those of the traditional gold standard method MIF assay to determine the immunodominant protein combination with the highest sensitivity and specificity.Results Of the 50 serum samples from patients with Ct infection,44 were positive and 6 negative by MIF.The results of ELISA with the combination of immunodominant proteins Pgp3,CT694 and CT875 as antigens were 97.73％ (43/44) consistent to those of MIF assay.Of the 30 serum samples from GICA-negative clients,all were negative by MIF.Meanwhile,no antibody was detected against any of the immunodominant proteins Pgp3,CT694 and CT875 in any of the serum samples from GICA-negative clients.Conclusions The ELISA with the combination of immunodominant proteins Pgp3,CT694 and CT875 as antigens has good sensitivity and specificity for serological screening for Ct infection,and is simple to operate and easy to popularize.

Objective To evaluate the clinical application value of reflectance confocal microscopy(RCM) in the diagnosis of several common diseases manifesting as papules in children, including lichen nitidus, verruca planae, lichen striatus, milium, molluscum contagiosum and lichen pilaris. Methods A total of 579 children clinically characterized by papules were recruited into this study. RCM was used to observe lesions and perilesional normal skin. The RCM features of 6 diseases manifesting as papules were analyzed and compared. Results Based on RCM images, 236 patients were diagnosed with lichen nitidus, 70 with verruca planae, 123 with lichen striatus, 40 with milium, 53 with molluscum contagiosum and 57 with lichen pilaris. All the 6 diseases had typical RCM features. Concretely speaking, RCM images of lichen nitidus lesions showed infiltration of dense inflammatory cells and melanophages in enlarged dermal papillae. In RCM images of verruca planae lesions, cells in the granular and spinous layers were arranged in concentric circles, giving a rose cluster?like appearance. RCM images of lichen striatus lesions revealed focal swelling of stratum spinosum, absent or local liquifaction degeneration of basal cells, and clustering of a moderate number of inflammatory cells in the superficial dermis. In RCM images of milium lesions, well?circumscribed round or oval structures containing highly but nonuniformly refractive materials could be seen in the dermis. RCM images of molluscum contagiosum lesions showed intact cystoid structures containing highly refractive molluscum bodies. Lowly to moderately refractive cutin ? like materials were observed along with the dilation of hair follicle infundibula in RCM images of lichen pilaris lesions. In RCM images, the 6 diseases were distinguished mainly based on structural features(patterns and refractivity)of skin lesions shown by continuous vertical scanning. Conclusion RCM is of great value to the diagnosis of diseases manifesting as papules in children.

Objective To investigate the effect of up-regulation of miR-132 on expressions of angiopoietin-1 (Ang-1)/endothelium-specific tyrosine kinase receptor 2 (Tie2) in focal cerebral ischemia reperfusion injury of rats.Methods Forty adult healthy SD rats were randomly divided into sham-operated group,cerebral ischemia group,miR-132 mimic group and negative control group (n=10).The models of middle cerebral artery occlusion in the later three groups were established by using modified Longa suture method.Rats in the miR-132 mimic group and negative control group were injected miR-132 mimic 15 μg and negative control 15 μg via paracele.Rats in each group were sacrificed 24 h after ischemia,and the brain tissues were collected;the total infarct volumes were calculated by TTC staining.The mRNA expressions ofmiR-132,Ang-1 and Tie2 in ischemic cerebral cortex tissues were detected by real-time fluorescence quantitative PCR.The protein expressions of Ang-1,Tie2,CD31 and vessel endothelial growth factor (VEGF) in ischemic cerebral cortex tissues were detected by Western blotting.Results The total infarct volume in the miR-132 mimic group was (27.92±3.05) mm3,which was significantly smaller than that in the cerebral ischemia group and negative control group ([51.34±2.86] mm3 and [50.46±2.57] mm3,P＜0.05).The relative mRNA expression levels of miR-132,A ng-1,Tie2,and the protein expression levels of Ang-1,Tie2,CD31,VEGF in the ischemic cerebral cortex tissues of the miR-132 mimic group were significantly higher than those in the negative control group,cerebral ischemia group and sham-operated group (P＜0.05);and those in the negative control group and cerebral ischemia group were significantly higher than those in the sham-operated group (P＜0.05).Conclusion Up-regulation ofmiR-132 expression could improve the ischemic states of ischemic stroke in rats,which might be related to Ang-1/Tie2 increased expressions to promote angiogenesis in ischemic brain tissues.