Epithelial architecture is formed in tissues and organs when groups of epithelial cells are organized into polarized structures.The epithelial function as well as signaling across the epithelial layer is organized by polarity proteins.Epithelial polarity complexes form and regulate epithelial integrity.Polarity proteins establish and maintain cell polarity,regulate tight junctions and adherens junctions.Polarity proteins act as neoplasms suppressors in mammals and interact with oncogenes,disrupt cell polarity,induce tumorigenesis.Therefore,polarity mechanisms of epithelial cell could therapeutic targets for clinical therapy.

Objective To evaluate effectiveness of peripheral Mohs micrographic surgery for the treatment of extramammary Paget′s disease (EMPD). Methods A total of 28 patients with EMPD were treated with peripheral Mohs micrographic surgery. The depth and extent of tumor infiltration were evaluated before the surgery. One day before the surgery, 20% aminolevulinic acid hydrochloride was topically applied to determine and label surgical margins under a Wood′s lamp. After fluorescence-based localization, peritumoral skin tissues were resected and underwent frozen-section examination according to the protocol for Mohs micrographic surgery. Meanwhile, the tumor was resected. After surgery, patients were followed up every 3 - 6 months to detect local recurrence and metastasis. Results Of the 28 patients, 25 were male and 3 were female. Six patients each underwent 3 sessions of frozen-section examination, and 12 patients each received 2 sessions, with an average of 1.86 sessions for each patient. During the follow-up for 5 - 72 months, local recurrence occurred in 3 cases, and 1 patient died of tumor metastasis and uremia after 2 years of follow-up. Conclusion Peripheral Mohs micrographic surgery is a time-saving and effective treatment for EMPD.

Objective To establish the HPLC characteristic chromatogram of Shenshitong Granules. Methods The chromatographie fingerprints were obtained through Thermo Hypersil GOLD-C18 column (4.6 mm×250 mm, 5μm) with the gradient elution solvent system composed of acetonitrile-0.2% phosphoric acid (0-20 min, 5%→10% acetonitrile;20-40 min, 10%→12%acetonitrile;40-60 min, 12%→14%acetonitrile;60-90 min, 14%→20%acetonitrile;90-120 min, 20%→28%acetonitrile). The detective wavelength was set at 280 nm;the flow rate was 1.0 mL/min;the column temperature was maintained at 30 ℃;the analysis time was 120 min. Results The HPLC characteristic chromatogram was built on basis of 10 batches of Shenshitong Granules, including 27 common peaks which contain the characteristic peaks of 6 Chinese herbal medicines, such as Radix Salvia Miltiorrhizae, Herba Lysimachiae, etc. Conclusion The established HPLC fingerprint has high sensitivity and good repeatability, and can be available for quality evaluation of Shenshitong Granules.

Objective To investigate the clinical features and prognostic factors in children with idi-opathic pulmonary arterial hypertension(IPAH). Methods The data of children with IPAH diagnosed in Beijing Children′s Hospital from January 2006 to December 2015 were collected. The cases were divided into survival group and death group according to the prognosis. The risk factors influencing prognosis in children with IPAH were identified by Logistic regression analysis. Results Total 26 patients were enrolled in this study,and 17 of them survived and 9 of them were dead. IPAH was found to be more common in school-age children. The average age of the patients was(6. 6 ± 3. 6)years and the male to female ratio was 1. 17∶1. All of the children had the symptoms with decreased activity tolerance and fatigue. The incidence of syncope and dyspnea and edema of lower extremity were 34. 6%,23. 1%. Accentuated pulmonic second sound(P2) was detected in 92. 3% of patients during physical examination, which was also the most common sign. About 50. 0% patients were functional class Ⅲ-Ⅳ. Echocardiography showed that all childrens′ mean pulmonary artery systolic pressure(PASP) was(79. 0 ± 20. 3) mmHg(1 mmHg=0. 133 kPa) and 16 of them with right ventricular dilatation. Electrocardiogram with ST-T changes and right ventricular high voltage were common. The average plasma level of brain natriuretic peptide(BNP) in all cases was(870. 4 ± 720. 9) pg/ml. The single factor analysis illustrated that syncope(OR=26. 25,95%CI 3. 04 -226. 60,P=0. 003),heart func-tional class Ⅲ-Ⅳ(OR=19. 199,95%CI 1. 88 -196. 53, P=0. 0128), PASP≥70 mmHg(OR =9. 936, 95%CI 1. 81-∞,P=0. 005),BNP≥850 pg/ml(OR=59. 991,95%CI 4. 69-767. 62,P=0. 002) indica-ted the worse outcome. Conclusion IPAH in children is rare and the prognosis is poor. Syncope, cardiac function grade,PASP and the levels of BNP are the main important factors affecting the prognosis of children with IPAH. BNP has important clinical significance for the evaluation of IPAH and the prognosis of the disease.

Objectives To assess the effect of nicorandil and trimetazidine on myocardial microcirculation reperfusion in patients with NSTEMI after elective PCI. Methods 80 patients with NSTEMI were randomly assigned to four groups: normal medicine group (CON), nicorandil group (NIC), trimetazidine group (TMZ) and Combination group (NIC+TMZ). The coronary angiography and PCI were performed after 10 days. MCE was taken since 72 hours after operation. Results There were no significant differences in baseline characteristics between the four groups (P > 0.05). The A, β and A ·β of group NIC were significantly higher than group CON (P < 0.05). There were no significant differences between the group TMZ and the group CON (P>0.05). The A,βand A·βof group NIC+TMZ were significantly higher than group NIC or TMZ (P<0.05). Conclusions Nicorandil can improve the situation of myocardial reperfusion after elective PCI in patients with NSTEMI, and it has some synergetic effect when combined with trimetazidine.

Purpose To assess the feasibility of dual-energy dual-source virtual noncontrast (VNC) CT in the diagnosis of laryngeal carcinoma,in order to reduce radiation hazard.Materials and Methods Forty-nine patients with clinically identified laryngeal carcinoma underwent conventional non-contrast (CNC) scan as well as arterial and venous dual-phase and dual-energy VNC scan.The VCN images of arterial and venous phase were obtained by using the dual-energy software.The mean CT value,signal-to-noise ratio (SNR),contrast noise ratio (CNR),lesion morphology,image quality and radiation dose were compared between the VNC and CNC images.Results The mean CT value,SNR,CNR and image quality scores of CNC were higher than those of VNC images of arterial and venous phase (P＜0.05).There was no significant difference in the image quality scores of VNC between the arterial and venous phase (P＞0.05).The image quality scores of VNC were all above 3,which could meet the diagnostic requirements.There was no significant difference in the shape,size,boundary and cystic necrosis of the lesions,invasion of the surrounding space,osteolytic destruction and size of the surrounding lymph node between the CNC and VNC image.The radiation dose of dual-phase dual-energy scan was 0.42 mSv,which was 18.3％ lower than that of the conventional three-phase scan.Conclusion For laryngeal carcinoma,the VNC images from the dual-energy CT scan can be used to reduce the radiation dose without affecting image quality and diagnosis accuracy,which has good clinical value.

Objective To establish a rapid,sensitive and specific diagnostic test for detecting Neisse-ria gonorrhoea.Methods The major outer membrane proteins(P Ⅰ )in different gonococcal serogroups were obtained by isolation of outer membrane complex with CTB-ethanol precipitation,the outer membrane proteins were extracted with Z 3,14 and EDTA,and purified with DEAE-Sepharose CL-6B to obtain P Ⅰ .Hybridoma cell lines producing McAbs against P Ⅰ were established with lymphocyte hybridoma techniques.Results The molecular weight of P Ⅰ A and P Ⅰ B were determined with SDS-PAGE as35.2kDa and36.7kDa,respectively.Five hybridoma cell lines producing McAbs continuouslly and stably against P Ⅰ A and P Ⅰ B were obtained,in-cluding two hybridoma cell lines producing McAbs against P Ⅰ A and three hybridoma cell lines producing McAbs against P Ⅰ B.The titers of McAbs in the supernatants in the cultures and in abdominal ascites of BALB/c were from1:64to1:256and from1:4096to1:16384,respectively;and the specificity of the McAbs against P Ⅰ A and P Ⅰ B was so high that they easily reacted with N.gonorrhoeae but did not with other antigens such as N.meningitidis etc.Conclusion The purified P Ⅰ and the McAbs obtained in the study provide a basis to establish a rapid,sensitive and specific diagnostic test for detecting N.gonorrhoea.

OBJECTIVETo detect sperm plasma membrane integrity (PMI) of cigarette smoking infertile males using SYBR-14/ PI fluorescent staining and flow cytometry and investigate its clinical significance.

METHODSWe collected semen samples from 132 cigarette smoking infertile men and 70 normal fertile controls, the former divided into a heavy-smoker group (> 20 cigarettes a day, n = 68) and a light-smoker group (< or = 20 cigarettes a day, n = 64). We performed computer-assisted semen analysis of the semen samples, and determined sperm PMI by flow cytometry after rinsing with PBS and staining by SYBR-14/PI, the sperm with normal PMI indicated as the percentage of those emitting green fluorescence (SYBR-14+/PI- %), dead sperm as the percentage of those emitting red (SYBR-14-/PI+), and moribund sperm as the percentage of those emitting both green and red (SYBR-14+/PI+).

RESULTSBoth the heavy- and light-smoker groups showed significant differences in SYBR-14-/PI+ % and SYBR-14+/PI- % from the normal controls (P < 0.01 or P < 0.05). SYBR-14+/PI- % was remarkably lower, while SYBR-14-/PI+ % markedly higher in the heavy-smoker than in the light-smoker group (P < 0.05). There was a significant correlation between SYBR-14+/PI- % and sperm motility (r = 0.938, P = 0.000).

CONCLUSIONSYBR-14/PI fluorescent staining and flow cytometry analysis could quickly and exactly detect sperm PMI. Cigarette smoking reduces sperm PMI and consequently sperm motility, which might be an important factor of male infertility.

Adult ; Case-Control Studies ; Cell Membrane ; pathology ; Flow Cytometry ; Humans ; Infertility, Male ; Male ; Semen Analysis ; Smoking ; adverse effects ; Spermatozoa ; cytology ; pathology

Objective: To evaluate the effects of repeated ultraviolet A (UVA) radiation on DNA damage, repair and replication processes in human skin fibroblasts, and to explore their mechanisms. Methods: Fibroblasts were isolated from the circumcised foreskins of 3 children in the Department of Urological Surgery, Third Affiliated Hospital, Sun Yat-sen University, and subjected to a primary culture. Cultured human skin fibroblasts of 3rd-10th passages were divided into 2 groups: UVA group treated with repeated UVA radiation to establish a chronic photodamaged cell model, and control group receiving no treatment. Cell counting kit 8 (CCK-8) assay, β-galactosidase staining and flow cytometry were performed to assess cellular proliferative activity, and determine the proportion of photoaged cells and apoptosis rate respectively. Reverse transcription reaction was performed to establish a differentially expressed cDNA library, which was then subjected to high-throughput sequencing. The cDNA sequencing results were compared between the control group and UVA group, and the differentially expressed genes were analyzed in Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The data were compared between the two groups by using two independent sample t test. Results: Compared with the control group, the UVA group showed significantly decreased cellular proliferative activity (72.0% ± 5.2% vs. 96.0% ± 3.7%, t = 6.51, P < 0.05) , but significantly increased proportion of photoaged cells (79.7% ± 5.2% vs. 6.4% ± 0.8%, t=24.12, P < 0.05) and apoptosis rate (29.0% ± 3.3% vs. 6.0% ± 5.9%, t= 5.89, P < 0.05) . Among the key enzymes involved in DNA mismatch repair, replication and base excision repair processes, the expression of DNA ligase 1 (Lig1) , ribonuclease (RNase) H2A and helicase Dna2 in the UVA group was 0.47 ± 0.13, 0.44 ± 0.07 and 0.49 ± 0.11 times (all P < 0.01) that in the control group respectively. After the UVA-induced chronic photodamage in the human skin fibroblasts, After the UVA-induced chronic photodamage in the human skin fibroblasts, Lig expression decrease could block DNA single-base excision repair, Lig and Lig1 expression decrease could block DNA multiple-base excision repair, Lig1 expression decrease could block DNA mismatch repair, and RNaseH2A, Dna2 and Lig1 expression decrease could block DNA replication. Conclusion Repeated UVA radiation can change the expression of key enzymes involved in DNA base excision repair, DNA mismatch repair and DNA replication processes in skin fibroblasts, and then affect DNA repair and DNA replication processes in skin fibroblasts.

OBJECTIVESTo study the fibrinolytic activity and the expression of uPAR and Annexin II in leukemic cell lines and their alterations on all-trans retinoic acid ( ATRA) treatment.

METHODSThe fibrinolytic activity was measured by chromogenic assay in NB4, SHI-1, K562, Jurkat and Raji cell lines. The protein expression of uPAR and Annexin II on cells surface and the mRNA expression of uPAR and Annexin II in cells of these cell lines were detected using flow cytometry and RT-PCR method respectively.

RESULTSThe plasmin activity in supernatant was increased significantly after incubation of SHI-1 and NB4 cells with plasminogen. The plasmin activity of NB4 cells was obviously decreased by ATRA. The plasmin activity of NB4 and SHI-1 cells was significantly decreased by uPAR monoclonal antibodies. The expressions of uPAR and Annexin II and their mRNA in SHI-1 and NB4 cells were higher than that in other cell lines. ATRA could remarkably decrease the expressions of Annexin II and uPAR and their mRNA in NB4 cells.

CONCLUSIONIn leukemia cell lines, NB4 and SHI-1 cells have stronger fibrinolytic activity. Both Annexin II and uPAR on the leukemic cell membranes might contribute to this activity. The high fibrinolytic activity can be corrected by ATRA by down-regulating Annexin I and uPAR mRNA and protein expression in NB4 cells.

Annexin A2 ; biosynthesis ; genetics ; Cell Line, Tumor ; Fibrinolysis ; drug effects ; Humans ; Leukemia ; metabolism ; pathology ; RNA, Messenger ; genetics ; Receptors, Cell Surface ; biosynthesis ; genetics ; Receptors, Urokinase Plasminogen Activator ; Reverse Transcriptase Polymerase Chain Reaction ; Tretinoin ; pharmacology