Objective:To investigate the changes in number,morphology and function of Langerhans cells and the molecular mechanisms in the tissues of condyloma accuminatum.Methods:CD1a,E-cadherin were measured by streptavidin peroxidase method and reverse transcriptase PCR in 26 CA lesions and 10 normal controls.Ultramicrostructure of LC was observed by transmission electronic microscope(TEM).Results:In CA tissues,the number of CD1a+ LC decreased(10.66?11.71,P

Objective To investigate the therapeutic efficacy and safety of carbon dioxide (CO2) fractional laser in treating periorbital syringoma.Methods Twenty-eight patients with syringoma were treated with ultrapulse CO2 fractional laser.Digital photos were taken before the treatment,1 month and 3 months after the treatment to evaluate the efficacy.The recovery process and side effects were also recorded to assess the safety.Dermatologists rated the efficacy in 0-4 points scales.The patients assessed overall satisfaction in 0-4 point scales.Results One month after the treatment,74.99 ％ of the patients had improvement rate of over 50 ％ and 67.85 ％ of the patients were satisfied or very satisfied with the overall efficacy.Three months after treatment,78.57 ％ of the patients had improvement rate of over 50 ％ and 71.42 ％ of the patients were satisfied or very satisfied with the overall efficacy.There was no statistically significant difference between 1 month and 3 months (P＞0.05).Conclusions Carbon dioxide fractional laser is an effective and safe approach to treat syringoma with minimized side effects.

Objective To evaluate the efficacy of the autologous epidermal grafting combining with NB-UVB radiation on the treatment of stable vitiligo.Methods Autologous epidermal graft acquired by suction blister under negative pressure were transplanted onto the laser-abraded depigmented areas,then the narrow-band UVB radiation was given after grafting to 35 patients with stable vitiligo.The onset time and the area repigmentation of every epidermal graft 3 months latter were observed (digital image contrasted before and after treatment).The results were compared with 37 cases only treated with autologous epidermal grafting.Results Both onset time and the cure rate in combined treatment group were statistically significant (P＜0.05) as compared with control group.Conclusions The combination of autologous epidermal grafting with NB-UVB radiation can shorten the onset time of repigmentation and promote generation and extension of the melanin.

In view of the deficiency of class hour and the limitations of classroom teaching in the course of ocular fundus disease, a network teaching platform, based on Browser/Server structure, was explored and constructed to assist classroom teaching. The network platform was constituted with teaching demonstration system, communication-test system, search system, and help system, mainly including 18 functional modules of learning-world et al. The students can be guided with the modules such as navigation, acting as a self-regulated learner through the modules such as learning-world, searching and downloading the related learning materials through the modules such as searching and discussing different learning topics with other students and their teachers through the modules such as forum. The network platform was used for classroom teaching of ocular fundus disease in auxiliary , and the result showed it was helpful to breaking the time and space constraints in conventional teach-ing, expanding the teaching content, solving the difficulties in teaching, improving the students' learning initiative, and realizing the interactive teaching.

AIM: The activity and expression of neutral endopeptidase (NEP) and the adrenomedullin (ADM) contents in various tissues were observed in septic shock and control rats to study the possible role of NEP in the change of ADM contents in tissues during septic shock. METHODS: The septic shock model of rats were established by cecal ligation and puncture (CLP). ADM contents, NEP activities, level of NEP mRNA and NEP protein were measured. RESULTS: (1) In early septic shock (ES), the ADM contents were generally higher in detected tissues, the NEP activity in left ventricle and small intestine were lower and was higher in blood than those in controls, and in lung, kidney and aorta were similar with the controls. NEP immunoreactive staining were less in lung, left ventricle, endothelium and media of aorta, but more in adventitia of aorta and kidney than those of the controls; (2) In late septic shock (LS), the ADM contents in small intestine was less but in plasma and other tissues were higher, and the NEP activity were less in plasma and other tissues than those in ES. The NEP immunoreactive staining were less in heart, endothelium and media of aorta, lung and kidney than those in ES, and was no significant change in adventitia of aorta compared with those of ES. RT-PCR found that NEP gene expression were significantly less in left ventricle, aortas, lung and small intestine than those in the controls. CONCLUSIONS: In septic shock rats, the NEP activity changes heterogeneously but the ADM contents elevate in most tissues. These results indicate that during the septic shock, the local concentrations and actions of ADM in various tissues may be regulated differently by the NEP. [

] AIM: Lysophosphatidic acid (LPA) is a bioactive phospholipid known to have growth factor-like activity on fibroblasts, and is involved in cardiovascular diseases. Besides direct effects, usually, LPA can work together with other bioactive factors to regulate cardiovascular homeostasis by induction of their expression and production, or increase in their activity. Among variety of bioactive factors, adrenomedullin (ADM) is a multifunctional peptide with an important cytoprotective effect against cardiovascular damage, but the interaction between ADM and LPA on adventitia remains unknown. METHODS: The experiment was performed on the bath of isolated rat aortic adventitia, ADM produced and secreted from adventitia stimulated by LPA was detected by using radioimmunoassay, proliferation in adventitia cells was evaluated by the level of [3H]-thymine incorporation, and prepro ADM gene expression was measured by semi-quantitative reverse transcriptase polymerase chain reaction. RESULTS: It was found that LPA stimulated aortic adventitia to secrete ADM and express its mRNA in a concentration-dependent manner. ADM inhibited LPA-induced proliferation in adventitial cells, and attenuated the activity of mitogen activated protein kinase (MAPK) stimulated by LPA. In contrast, the treatment with specific antagonists of ADM receptor potentiated the LPA-induced proliferation in adventitial cells. CONCLUSION: LPA stimulates adventitia to produce and secrete ADM, and in turn, ADM produced by adventitia regulates the vascular biological effects of LPA. [

Objective To establish a model for preparation of tissue-engineered skin grafts with hTERT cells carrying human papillomavirus type 6 (HPV 6) genome in vitro, so as to lay a foundation for studying HPV life cycle. Methods The full-length linear HPV6 genome and plasmid pEGFP-▲EGFP were electrophoretically cotransferred into hTERT cells. After selection using G418 resistance, Southern blotting was performed to determine the viral load of HPV6 in transfected cells. 3T3 J2 trophoblastic cells, type I rat-tail collagen and hTERT cells containing the full-length HPV6 genes (HPV6.hTERT cells)were mixed and cocultured on metal meshes to form skin graft-like structures. Hematoxylin and eosin (HE)staining was performed to observe the structure of formed skin grafts, an immunohistochemical assay to measure the expression of HPV6 L1 protein, and electron microscopy to observe virus particles in the skin grafts. Results The linear HPV6 gene was successfully transferred into hTERT cells, and Southern blotting showed the presence of HPV6 DNA in the transferred hTERT cells. The HPV6.hTERT cells, which were cocultured with 3T3 J2 trophoblastic cells and type I rat-tail collagen, proliferated and differentiated over time, and gradually formed skin grafts giving the appearance of verrucous hyperplasia. HE staining showed that the cocultured HPV6.hTERT cells could form typical stratified structure of skin after 7 days of cultivation, and histopathologic features of HPV infection, including obvious papillomatous hyperplasia, presence of vesicular cells, hyperkeratosis and parakeratosis, could be observed after 21 days. The immunohistochemical assay showed the expression of HPV6 L1 protein in the upper portion of skin grafts, and electron microscopy revealed the presence of HPV6 virus particles in skin grafts. Conclusions The established model for preparation of tissue-engineered skin grafts using HPV 6 genome-carrying cells provides a basis for biological studies of HPV, but its application is limited to some degree.

Objective To investigate the correlation between serum vitamin D level and central precocious puberty(CPP)in girls.Methods A total of 103 girls(case group)with central precocious puberty from Ningbo Woman and Children's Hospital and 53 healthy girls(control group)from health check-ups in Ningbo Women and Children's Hospital were collected as subjects.The serum levels of 25-hydroxyvitamin D3[25(OH)D3]in the two groups were detected by chemiluminescence method.The weight and height of girls in the case group were measured.The serum levels of follicle-stimulating hormone(FSH)and its peak value,luteinizing hormone(LH)and its peak value,estradiol(E2),prolactin(PRL),human chorionic gonadotropin(HCG)and thyroid function were measured by radioimmunoassay.The peak value of LH/FSH was calculated.B ultrasound examination of uterine adnexa was completed to calculate uterine volume and bilateral ovarian volume.According to the results of serum 25-(OH)D3,girls in the case group were divided into normal vitamin D group and vitamin D deficiency group,and the differences of hormone levels,uterine and ovarian development between the two groups were compared.Results The serum level of 25-(OH)D3 in case group was lower than that in control group,and the vitamin D deficiency rate in case group was higher than that in control group,with statistical significance(P<0.05).The age of breast nodules in vitamin D deficiency group was lower than that in vitamin D normal group(P<0.05).There were no significant differences in body weight,height,body mass index(BMI),uterine volume and left ovarian volume between vitamin D normal group and vitamin D deficiency group(P>0.05),and the right ovarian volume in vitamin D deficiency group was significantly higher than that in vitamin D normal group(P<0.05).There were no significant differences in serum levels of FSH,LH,PRL,HCG,peak value of FSH and thyroid function between normal and deficient groups(P>0.05).The levels of E2,LH and LH/FSH in vitamin D deficiency group were significantly higher than those in vitamin D normal group(P<0.05);25-(OH)D3 was negatively correlated with LH/FSH peak(r=-0.197,P<0.05),but was not significantly correlated with thyroid function,FSH,LH,PRL,E2,HCG,FSH and LH peak(P>0.05).Conclusion Vitamin D deficiency is associated with central precocious puberty in girls.Vitamin D deficiency may lead to early onset of precocious puberty.Vitamin D deficiency may affect the hypothalamic-pituitary-gonadal axis function,resulting in changes in reproductive hormone indexes and consequent increase in ovarian volume in girls.

Objective: To establish a simple method for isolation, purification and cultivation of primary retinal microvascular pericytes (RMPs) from mice. Methods: Retinas were isolated from mice following with mechanical morcel, enzymatic digestion and filtration.The retinal fragments were incubated with low glucose DMEM with 20% fetal bovine serum after 24 hours pre-incubation.Differential digestion was used for purification of primary RMPs.Morphological examination of cells was performed by phase contrast microscopy, and further characterization was analyzed by immunocytochemistry.Functional assay was evaluated by the pericytes-endothelial cells (ECs) co-culture system.The treatment and use of experimental animals followed the regulations on the administration of experimental animals promulgated by the state science and technology commission. Results: Cells migrated out of fragments after 24 hours of incubation, and developed into small or large colonies gradually.The cells and their subpassages presented typical pericyte morphology with large irregular triangular cell bodies and multiple long processes.No contact inhibition was observed.Most cells uniformly expressed the cellular markers α-smooth muscle actin (α-SMA) and platelet-derived growth factor receptor-β (PDGFR-β), a few cells expressed the cellular markers glial fibrillary acidic protein (GFAP), but no cell expressed von Willebrand factor (vWF). The purity rate of RMPs was up to 97%.In the co-culture system, RMPs directly contacted with ECs to form the capillary-like cords in vitro. Conclusions A simple method for the isolation, purification cultivation of mouse RMPs is established, and active RMPs can be readily obtained by this method.