Objective To explore the value of evaluating rat liver fibrosis using contrast enhanced sonography.Methods Carbon tetrachloride was used to induce liver fibrosis.Under the mode of pulse-inversion harmonic imaging,ultrasound contrast agent was bolus iniected through tail vein,hepatic artery arrival time(HAAT),portal vein arrival time(PVAT),hepatic vein arrival time(HVAT)was recorded,and their relationship with immunohistochemical results were analyzed.Results There was no statistical difference of HAAT,PVAT between two groups among S0-S4 stage.HVAT was shorter in S3 and S4 stage than in S0-S2 stage(P＜0.05).HVAT was negative correlated with the percentage of immunohistochemical positive area of C-Ⅳ,LN,the correlation coefficients were-0.680 and -0.639 respectively.Conclusions Contrast enhanced sonography is useful for the diagnosing liver fibrosis.

Objective To investigate the expression of ezrin in seborrheie keratosis(SK),basal cell carcinoma (BCC).squamous cell carcinoma (SCC)and its relation to elinical pathology parameters.Methods Skin samples were collected from 36 patients with SCC,27 patients with BCC,20 patients with SK and 10 normal human controls.Immunohistochemistry was performed to detect the expression of ezrin in these samples.The relationship between ezrin expression and prognosis of SCC was assessed using COX regression analysis.Results The positivity rate of ezrin in SK,BCC and SCC samples was 25.0%,66.7%,91.3%,respectively,compared to 20.0%in normal controls.Compared with the controls,a significant increase was observed in the expression of ezrin in patients with BCC and SCC(both P＜0.05).but not in those with SK(P＞0.05).Moreover.increased expression level of ezrin was correlated with a high degree of tumor malignancy,advanced pathological stage,and occurrence of lymph node metastasis of SCC (r=0.87,0.80,0.89,respectively,all P＜0.01).COX regression analysis revealed that the expression level of ezrin was an independent factor for the prognosis of SCC.Conclusion The expression level of ezrin is closely related to the malignancy of skin tumors,pathological grading and lymph node metastasis of SCC.

BACKGROUND:Anterior cruciate ligament (ACL) injury is a commonly sport-induced knee joint injury that does serious harm to the knee stability. ACL reconstruction is a commonly used treatment method, but researches on 1/2 peroneus longus tendon (PLT) graft are rarely reported. OBJECTIVE: To investigate the clinical outcomes of removing the autologous ipsilateral 1/2 PLT under arthroscopy for ACL reconstruction. METHODS:106 patients with complete ACL rupture in the Affiliated Hospital of Traditional Chinese Medicine, Southwest Medical University from December 2010 to December 2014 were enrolled, and autologous ipsilateral 1/2 PLT was removed under arthroscopy for ACL reconstruction. At baseline, 3, 6 and 12 months postoperatively, the knee stability was evaluated manually through the anterior drawer test, Lachman test, and pivot-shift test, and the knee function was evaluated by Tegner activity scale, Lysholm and International Knee Documentation Committee scores. RESULTS AND CONCLUSION: Postoperative anterior drawer test, Lachman test, and pivot-shift test tests were negative in all patients. In terms of Tegner activity scale, Lysholm and International Knee Documentation Committee scores, there were significant differences at baseline and postoperative 3 months as compared with postoperative 6 months (P < 0.05); the scores at baseline and postoperative 3 months showed significant differences compared with 12 months postoperatively (P < 0.05); the scores showed no significant difference between 6 and 12 months postoperatively (P > 0.05). These results indicate that autologous ipsilateral 1/2 PLT is a good choice for ACL reconstruction under arthroscopy, achieving rapid and satisfactory functional recovery of the knee joint, which is not only minimally invasive and easy to operate, but also exhibits good therapeutic efficacy.

OBJECTIVETo study the promoter methylation pattern of p16 and hMLH1 genes in esophageal squamous cell carcinoma and reflux esophagitis, and to correlate the results with clinical and pathologic findings.

METHODSTwelve cases of normal esophagus, 13 cases of esophageal squamous cell carcinoma, 43 cases of reflux esophagitis with basal cell hyperplasia and 21 cases of reflux esophagitis with dysplasia, as confirmed by endoscopic and pathologic examination, were enrolled into the study. Genomic DNA was extracted. The promoter methylation status of p16 was measured by methylation-specific polymerase chain reaction. The promoter methylation status of hMLH1 was measured by sodium bisulfite-restriction enzyme digestion. Immunohistochemical study for p16 and hMLH1 proteins was also carried out.

RESULTSThe rates of p16 methylation in normal esophageal epithelium, basal cell hyperplasia, dysplasia and esophageal squamous cell carcinoma were 0/12, 14.0% (6/43), 38.1% (8/21) and 6/13, respectively. The p16 methylation correlated with the progress of esophageal lesions. On the other hand, the hMLH1 methylation was not observed in the normal esophageal epithelium and reflux esophagitis. One case of esophageal squamous cell carcinoma showed the presence of hMLH1 methylation. The hMLH1 promoter hypermethylation did not correlate with the clinical and pathologic features.

CONCLUSIONSThe p16 methylation may be one of the earliest events in the pathogenesis of esophageal squamous cell carcinoma and is also observed in reflux esophagitis. Reflux esophagitis may be related to the development of esophageal squamous cell carcinoma in Chinese population. In contrast, hMLH1 methylation may not be directly involved in the tumorigenesis of esophageal squamous cell carcinoma.

Adaptor Proteins, Signal Transducing ; genetics ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; DNA Methylation ; Esophageal Neoplasms ; genetics ; pathology ; Esophagitis, Peptic ; genetics ; pathology ; Esophagus ; pathology ; Female ; Genes, p16 ; Humans ; Hyperplasia ; Male ; Middle Aged ; MutL Protein Homolog 1 ; Nuclear Proteins ; genetics ; Precancerous Conditions ; genetics ; pathology ; Promoter Regions, Genetic ; genetics

Objective: To analyze the duration and influencing factors of moderate and vigorous physical activity(MVPA) on weekends for primary school students in grades 4 to 6 in Beijing, and to provide a reference for formulating health education and promotion measures. Methods: Multi stage stratified cluster sampling method was used to randomly select 2 515 students from grades 4-6 in 14 primary schools in Beijing, and a self administered questionnaire was used to record MVPA on weekend, social demographic characteristics, other related health behaviors and knowledge. Multivariate Logistic regression analysis was used to explore the influencing factors of MVPA on weekends. Results: The prevalence of insufficient MVPA on weekends in Beijing was 63.54 %, and the prevalence was higher among girls (69.92%) than boys (57.81%) ( χ 2=39.65, P <0.01). Multiple Logistic regression analysis revealed that girls ( OR =1.74), living in rural areas ( OR =1.41), participants attending general schools ( OR = 1.34 ), from divorced family ( OR =1.46), and short sleep duration ( OR =1.50) were more likely to fail to meet the MVPA recommendations( P <0.05). Conclusion It is quite common that no sufficient weekend MVPA among senior primary school students, among them, the outer suburbs and schools with relatively weak teaching resources are the key places that need attention, and girls are the key groups that need attention.

Background Erythropoietin (EPO) has a protective effect on retinal neurons in many retinal diseases,but regarding the effect of EPO on apoptosis of retinal photoreceptor cells in retinal detachment (RD) is uncompletely clear.Objective This study was to investigate the protective effect of endogenous EPO on photoreceptors in a rat model of RD and explore its possible mechanism.Methods Seventy-two Sprague- Dawley (SD) rats were randomly assigned to control group,RD group,RD+PBS group,RD+erythropoietin soluble receptor (EPOsR) 2, 20, 200ng groups with 12 rats for each group.1.4% hyaluronic acid was slowly injected into the subretinal space to induce RD in rats,and PBS or 2,20 or 200ng EPOsR was then injected into the vitreous space.On day 3 after RD,apoptotic photoreceptors were detected using transferase-mediated dUTP nickend labeling (TUNEL),and caspase-3 activity was assessed by Western-blot and immunofluorescence staining.On day 14 after RD,retinal histopathologic examination was carried out and outer nuclear layer (ONL) thickness was measured under the light microscope.The use of animals complied with the Statement of Association for Research in Vision and Ophthalmology. Results Apoptotic photoreceptors were seen in ONL of rats of the RD group.Apoptotic photoreceptors were gradually increased with the elevation of EPOsR dose in the vitreous cavity.Western blot and immunofluorescence consistently showed that the gray scale of caspase-3 activity was 0.15±0.04,0.49±0.03,0.50±0.07,0.63±0.03,0.69±0.04 and 0.83±0.04 in the normal group,RD group,RD +PBS group,RD+EPOsR 2,20,200ng groups respectively with statistically significant differences (F=76.016;P=0.000),and caspase-3 activity was considerably stronger in the RD+EPOsR 200ng group than the other groups (P＜0.01).On day 14 after RD,the ONL thicknesses in the normal control group,RD group,RD+PBS group,RD+EPOsR 2,20,200ng groups were (47.39±3.39)μm,(33.96±3.54)μm,(31.83±5.21)μm,(31.40±2.63)μm,(24.99±2.06)μm and (19.30±3.71)μm,showing significant differences among these groups (F=44.733,P=0.000).ONL thicknesses the groups treated with different doses of EPOsR were markedly thinner than that of the RD group and RD +PBS group (P＜0.01).Conclusion EPOsR induces apoptosis of retinal cells and enhances the activity of caspase-3 in a dose-dependent manner.Endogenous EPO can protect photoreceptors against anoxia-mediated damage in RD eyes through decreasing caspase-3 activity and inhibiting apoptosis.

Objective: To explore the prevalence and associated factors of non learning-based screen time of 4-6 grade school students in Beijing, and to provide a basic data for further research on hazard control measures such as myopia, overweight and obesity among children and adolescents in Beijing. Methods: Multistage stratified random cluster sampling method was adopted. A total of 2 515 primary school students were randomly selected in from schools in Beijing, self-developed questionnaire was used to investigate on the time for each electronic products usage for non-learning purpose, the total of electronic products usage every day and other information. Results: The rates of screen time >15 minutes each time and ≥1 h daily were 48.43%, 22.90%, respectively. Multiple Logistic regression analysis showed that ordinary school, 6 th grade, male, single-parent family and other types of family, and lack of moderate-to-vigorous physical activity at weekend were associated with the students’ non learning-based screen time (OR=1.66, 2.28, 1.27, 1.44, 1.87, 2.20, P<0.05). Conclusion The situation of excessive screen time of primary school students was improved in Beijing, but still be prevelent, male students of grade 4 should be given more attention, and more importance should be attached to the offect of family in children.

Objective To retrospectively analyze the therapeutic effect on patients with tonsillar carcinoma and factors affecting their prognosi. Methods Clinical data of 61 patients pathologically confirmed with tonsillar carcinoma without distant metastasis were analyzed. All the patients were treated in Cancer Center of Sun Yat-sen University from April 1997 to April 2008. There were 2 patients with undifferentiated carcinoma, 26 with poorly differentiated squamous cell carcinoma and 33 with median-well differentiated squamous cell carcinoma. According to the AJCC 2002 staging criteria for head-neck cancers, there were 9 staged Ⅰ cases, 7 staged Ⅱ cases,23 staged Ⅲ cases and 22 staged Ⅳ cases. The treatment was radiotherapy alone in 27 cases, radiotherapy combined with chemotherapy in 23 cases, surgery combined with postoperative radiotherapy in 6 cases, neoadjuvant chemotherapy plus surgery combined with postoperative radiotherapy in 3 cases, radiotherapy with salvage surgery in 2 cases. Results The overall 5-year survival rate was 50. 2%. For 16 cases with staged Ⅰ-Ⅱ staged, there were 8 cases with radiotherapy alone, 5 years survival was 50. 0%, 6 cases with surgery combined with postoperative radiotherapy, 5 years survival was 83.3%. The difference between the two treatments was not significant in statistics (P=0.318). For Ⅲ-Ⅳ staged 45 cases, there were 19 cases with simple radiotherapy, 5 years survival was 51.5%, 21 cases with radiotherapy combined with chemotherapy, 5 years survival was 36.4%, 5 cases with surgery combined with postoperative radiotherapy, 5 years survival was 75.0%. The difference among the three treatments was not significant in statistics (P=0.239). According to T stages, the 5-year survival rates of stage T1-T4 cases were 91.8%, 46.8%, 29.1%, 0% respectively (χ~2=30.168, P<0.001). Multivariate analysis demonstrated that T stage, therapeutic effect of primary site and cervical metastatic lymph node were the independent prognostic factors (P<0.05). Conclusions T stage, the therapeutic effect of primary site and cervical metastatic lymph node were the independent prognostic factors. For Ⅰ-Ⅱ staged tonsillar tumor cases, based on organ preservation, were tendency to choice simple radiotherapy. For Ⅲ-Ⅳ staged cases,yet the ralationships between therapeutic mode and therapeutic effect still need further researches.

In order to enhance the anticoagulant properties of decellularized biological materials as scaffolds for tissue engineering research via heparinized process, the decellularized porcine liver scaffolds were respectively immobilized with heparin through layer-by-layer self-assembly technique (LBL), multi-point attachment (MPA) or end-point attachment (EPA). The effects of heparinization and anticoagulant ability were tested. The results showed that the three different scaffolds had different contents of heparin. All the three kinds of heparinized scaffolds gained better performance of anticoagulant than that of the control scaffold. The thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (APTT) of EPA scaffold group were longest in all the groups, and all the three times exceeded the measurement limit of the instrument. In addition, EPA scaffolds group showed the shortest prepared time, the slowest speed for heparin release and the longest recalcification time among all the groups. The decellularized biological materials for tissue engineering acquire the best effect of anticoagulant ability in vitro via EPA heparinized technique.
Animals ; Anticoagulants ; chemistry ; Biocompatible Materials ; chemistry ; Heparin ; chemistry ; Liver ; Swine ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo observe the effect of recombinant adenovirus Ad-ING4 on K562 cells.

METHODSHuman ING4 recombinant transfer vector pAdTrack-CMV-ING4 was constructed by enzyme digest and ligation of human ING4 gene which was obtained through site specific point mutation of mouse ING4. The vector was co-transduced into BJ5183 E. coli with pAdEasy-1. The new recombinant adenovirus vector pAdEasy-1-pAdTrack-CMV-hING4 was transfected into QBI-293A cells. To obtain the ING4 recombined adenovirus (Ad-ING4). Ad-ING4 was used to infect K562 cells. The effect on K562 cells of ING4 was tested by LSCM FCM and immunohistochemistry.

RESULTSHuman ING4 recombinant adenovirus vector was constructed successfully, and high titre ING4 recombinant adenovirus (Ad-ING4) was obtained. ING4 can down-regulate the expression of bcl-2 and up-regulate expression of bax. The apoptosis of K562 cells induced by ING4 was proved by LSCM FCM and immunohistochemistry. The apoptosis rate was 19.7% (after 72h), which displayed significant difference compared with that of control groups (P < 0.01).

CONCLUSIONAd-ING4 can inhibit the growth of K562 cells and induce the cells apoptosis. The human ING4 recombinant adenoviral vector constructed might provide an approach to the target therapy of tumors.

Adenoviridae ; genetics ; Animals ; Apoptosis ; genetics ; Base Sequence ; Carrier Proteins ; genetics ; Cell Cycle Proteins ; genetics ; Cell Proliferation ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; K562 Cells ; Mice ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Plasmids ; genetics ; Transfection ; Transformation, Bacterial ; Tumor Suppressor Proteins ; genetics