Background Sulforaphane (SFN) is an effective chemopreventive agent and can regulate the biological molecular mechanisms to inhibit the overgrowth of cells.Autophagy is a biological process of maintaining cellular internal environment.Understanding the affection of SFN to biological behavior of human lens epithelial cells (LECs) and the association of SFN with autophagy is helpful for the prevention and target treatment of posterior capsule opacification (PCO).Objective This study was to investigate the eradication effeccts of SFN on residual lens cell population in vitro posterior capsule opacification (PCO) model and evaluate the mechanism of SFN-induced cell death.Methods In vitro human capsular bag models were generated from fresh donor eyes by phacoemulsification and were cultured in EMEM containing 2％ fetal bovine serum (FBS).Different concentrations of SFN (0,1,10 and 100 μ mol) were added in the medium for 30 days respectively according to grouping,and the growth of LECs was observed by optical microscope and immunofluorescence technique.FHL124,a human LEC line,was cultured with EMEM containing 5％ FBS and divided into 0,1,10,30 and 100 μmol SFN groups.Lactate dehydrogenase (LDH) release rate in the medium was detected to evaluate cell damage/death.The migration of the cells on capsular bags was assessed by scratch test.The ultrastructure and number of autophagosomes were examined under the transmission electron microscope.The expression of LC3 in the cells were detected using Western blot in the presence or absence of autophagy inhibitors.Results The cell coverage rates on the capsular bags were significantly lower in the 10 and 100 μ mol/L SFN groups than those in the 0 and 1 μmol/L SFN groups,with a statistically significant difference among the groups (F =48.57,P ＜ 0.01).Immunofluorescence showed that the density of F-actin-and Vimentin-positive cells was evidently decreased in the 10 and 100 μmol/L SFN groups compared with 0 and 1 μ mol/L SFN groups.The releasing levels of LDH (absorbancy) were 0.19± 0.03,0.39±0.06,0.56±0.07,0.68±0.08 and 0.89±0.09 in the 0,1,10,30 and 100 μ mol/L SFN groups,respectively,and the releasing level of LDH was gradually increased in the 10 and 100 μ mol/L SFN groups in comparison with the 1 μmol/LSFN group (all at P＜0.01).With the increase of SFN concentration,the reduction rate of scratched area decreased with the increase of SFN concentration,and the decrease of scratch area was significantly lower than that of adjacent low mass concentration group and the differences were statistically significant (P＜0.05).The relative expressions of LC3-Ⅱ protein were 0.423±0.003,14.543±0.024,0.668±0.024 and 0.576±0.056 in the blank control group,SFN group,SFN + 3-MA group and 3-MA group,respectively,and the relative expressions of LC3-Ⅱ protein were significantly lower in the SFN+3-MA group and 3-MA group than those in the SFN group (all at P＜0.01).The number of autophagosomes was 4.07±0.32,4.13±0.34,9.21 ±0.53 and 21.02± 1.34 in the blank control group,and 1,10,100 μmol/L SFN groups,and the number of autophagosomes in the 10 and 100 μ mol/L SFN groups was significantly higher than that in the blank control group and 1 μmol/L SFN group (all at P＜0.01).Conclusions SFN mediates LECs death by promoting autophagy in ex vivo capsular bags,and SFN may be a novel agent of potential chemopreventive and target treatment for PCO.

Objective To explore the feasibility of applying small private online course (SPOC) in the course of Microcomputer Principle and Interface Technology to solve its problems in complicated knowledge points,abstract contents and difficulty in understanding.Methods The characteristics of the undergraduate students in the military medical university and SPOC mode were analyzed,and then the design and implementation of a SPOC-based Microcomputer Principle and Interface Technology course were explored in the military medical university.Results By applying the SPOC into the teaching of Microcomputer Principle and Interface Technology course,the student could find out the forgotten or leaked knowledge and reiterate them to reinforce the memory of those knowledge points,which promoted their self-regulation of learning.Besides,teaching the students with real cases not only increased the learner's enthusiasm but also strengthened the military medical background.The offline group discussion facilitated the students in understanding and application of knowledge.Conclusion Applying SPOC mode into PMIT teaching no doubt improves the effect and quality of teaching and learning.

Objective To determine the three-dimensional (3D) texture features extracted from intensity and high-order derivative maps that could reflect textural differences between bladder tumors and wall tissues,in order to achieve bladder cancer and wall tissue identification.Methods A total of 62 cancerous and 62 wall volumes of interest (VOI) were extracted from T2-weighted MRI datasets of 62 patients with pathologically confirmed bladder cancer.To reflect heterogeneous distribution of tumor tissues,3D high-order derivative maps (the gradient and curvature maps) were calculated from each VOI.Then 3D Haralick features based on intensity and high-order derivative maps and Tamura features based on intensity maps were extracted from each VOI.Statistical analysis was proposed to first select the features with significant differences and then obtain a more predictive and compact feature subset to verify its differentiation performance.Results From each VOI,a total of 58 texture features were derived.Among them,37 features showed significant inter-class differences (P≤ 0.01).Conclusion The results suggest that 3D texture features deriving from intensity and high-order derivative maps can reflect heterogeneous distribution of cancerous tissues.

Objective To establish a method of PKH26 labeled bone marrow-derived endothelial progenitor cells (EPCs) into rats with liver fibrosis and observe cell immigration and differentiation in the liver after transplantation.Methods Bone marrow-derived EPCs were isolated and cultured from rats with liver fibrosis,and then labeled with PKH26 in vitro.Under the scanning confocal microscopy and flow cytometry,PKH26 fluorescent labeling rate and cell survival rate were measured.EPCs of PKH26 fluorescent labeling were transplanted into rats with liver fibrosis via the tail vein,and the migration situation was observed in the liver.Endothelial cell markers CD31 and von willebrand factor (vWF) were detected by using immunofluorescence.Results The PKH26-labeled EPCs appeared red fluorescence and the labeling rate was 96.65 ％.As compared with unlabeled cells,the labeled cells grew well,and had no significant changes in the growth curve.After transplantation into the liver of rats,the PKH26 labeled cells were mainly distributed in blood vessel endothelium along fibers and hepatic sinusoids in hepatic lobule.Endothelial cell-specific antigens such as CD31 and vWF could be detected along the vascular walls.Conclusion PKH26 could be used to label and track EPCs in the liver of rats with liver fibrosis in vitro.The PKH26-labeled cells may migrate to the surrounding of the hepatic vessels and differentiate into mature endothelial cells.

BACKGROUND:Theoretically, bone marrow-derived endothelial progenitor cells (EPCs) from liver fibrosis rats could be filtered by the pathological environment in vivo. These EPCs would be more adapted to the micro-environment of liver fibrosis, and easier to differentiate into mature endothelial cells participating in the intrahepatic vascular remodeling after transplanted into the liver.OBJECTIVE:To explore the effectiveness of transplantation of bone marrow-derived EPCs from the liver fibrosis environment in liver fibrosis rats.METHODS:Twenty-eight Wistar rats were randomly divided into three groups as follows:normal group (n=8) were injected with olive oil, twice per week; model group (n=10) were infused with carbon tetrachloride at a dose of 3 mL/kg body weight (double doses for the first time), twice per week, and infused with normal saline through the tail vein at 2, 3 and 5 weeks; EPCs transplantation group (n=10) were infused with carbon tetrachloride at a dose of 3 mL/kg body weight (double doses for the first time), twice per week, and infused with EPCs suspension through the tail vein at 2, 3 and 5 weeks. Six weeks after final injection, the angiogenesis, hepatocyte proliferation and pathological changes in the liver tissues were observed. The liver function and coagulation function were tested.RESULTS AND CONCLUSION:(1) The pathological changes of the liver:in the model group, fatty degeneration and hepatocyte necrosis in the liver tissue were serious, inflammatory cells were infiltrated around the portal and central vein,the portal areas expanded, and fibrous tissues overgrew. Compared with the model group, these changes were significantly relieved in the EPCs transplantation group (P < 0.05). (2) The expressions of liver-related proteins:compared with the normal group, the levels of hyaluronic acid, laminin, type III procollagen, vascular endothelial growth factor, epidermal growth factor were significantly increased in the model group (P < 0.05). Compared with the model group, the levels of hyaluronic acid, laminin and type III procollagen were decreased significantly (P < 0.05), and the levels of vascular endothelial growth factor and epidermal growth factor were increased in the EPCs transplantation group (P < 0.05). (3) Liver function and coagulation function:compared with the normal group, the liver function and blood blotting function of rats were seriously damaged in the model group (P < 0.05). Compared with the model group,the liver function and coagulation function were obviously improved in the EPCs transplantation group (P < 0.05). To conclude, transplantation of bone marrow-derived EPCs from the liver fibrosis environment is effective for liver fibrosis in rats. The mechanism may be associated with the promotion of angiogenesis in the liver.

Objective To evaluate the effect and pathological characters for patients with early esophageal carcinoma and intraepithelial neoplasia after endoscopic submucosal dissection (ESD). Methods 69 patients from January 2013 to January 2016 were treated with ESD at the early stage of esophageal carcinoma and intraepithelial neoplasia. The clinical features and the size of the lesions of all the patients were collected. Then analyzed postoperative complications and pathological characteristics. Results Among 69 cases, 16 were early esophageal cancer, 38 were high-grade esophageal neoplasia and 35 were low-grade esophageal neoplasia. The whole piece resection rate was 100.00 % (69/69), complete resection rate and curative resection rate was 95.65 % (66/69), respectively. The largest removal diameter is 7.0 cm. Biopsy accuracy was 69.57 % (48/69). Compared with biopsy, diagnostic accuracy with ESD specimens is higher. Conclusion The early esophageal carcinoma and intraepithelial neoplasia can be treated with ESD. ESD can resect lesions primarily, provide complete specimen for further pathological assessment and improve diagnostic accuracy.

Objective To investigate the effects of CD147/MMP-9 pathway on early left ventricular remodeling Methods 30 healthy eight-week male SHR were divided into 3 groups (n = 10 for each group). SHR group received tail vein injections of normal saline weekly; CD147 group received CD147 of 600 ng·kg-1 weekly; and CD147+DOX group received CD147 of 600 ng/kg weekly and intragastric administration of DOX ( doxycycline ) of 30 mg/kg daily . 10 healthy eight-week male Wistar-Kyoto rats (WKY group) were treated as SHR group. Echocardiography, myocardial sections microscopy examination (HE and VG stain), and Western blot (for assessing levels of MMP-9, TIMP-1, CD147, and collagen I and Ⅲin myocardial tissues) were performed on day 56. Left ventricular weight index (LVWI)was measured and calculated. Collagen volume fractions (CVF) were obtained by image analysis. Results As compared with WKY group , levels of CD147 , MMP-9 , and MMP-9/TIMP-1 were lower but TIMP-1 and collagenⅠand Ⅲ were significantly higher in SHR group. The abundance of CD147 and MMP-9 protein and the ratio of MMP-9/TIMP-1 were obviously increased in CD147 group than in SHR group (P < 0.05). Levels of CD147, MMP-9, and MMP-9/TIMP-1 did no differ between CD147+DOX group and CD147 group. LVWI and contents of collagenⅠand Ⅲ were obviously declined in CD147 group as compare with SHR group. Cardiomyocyte hypertrophy , partial myocardial fibre rupture , myocyte dissolution and fuzzy myocardial fibre boundaries , more abundant of collagen fibers, and higher CVF were found in SHR group. Cardiac fibrosis was significantly improved after CD147 intervention, but the action was suppressed as DOX was administrated simultaneously. Conclusions Early ventricular remodeling may be involved in the inhibition of CD147/MMP-9 pathway in SHR. Input of CD147 to upregulate the pathway can improve the remodeling.

Objective To investigate the expression and clinical significance of epithelial cell adhesion molecule(EpCAM) and Claudin-18 in gastric cancer.Methods Surgical specimens of gastric cancer were taken from 64 patients.The histological diagnostic criteria were based on WHO standards.Immunohistochemical staining was used to detect expression levels of EpCAM and Claudin-18 in all samples.The relationship between the expression of EpCAM and Claudin-18 and clinicopathological parameters of gastric cancer was analyzed.Correlations of the prognosis of gastric cancer patients with EpCAM and Claudin-18 expression were analyzed using the Cox proportional hazards regression model.Results The positive expression rate of EpCAM was elevated with the increase in tumor size and the occurrence of distant metastasis(x2 =4.526 and 36.090,P=0.033 and 0.000).There were significant differences in Claudin-18 protein expression among different age,TNM stage,growth pattern,depth of invasion and distant metastasis patients(all P＜0.05).Cox regression model analysis showed that tumor size and distant metastasis were the main risk factors affecting the survival of patients with gastric cancer (RR =50.076 and 1.617,P =0.016 and 0.032),while levels of EpCAM and Claudin-18 were not independent risk factors (both P ＞ 0.05).Conclusions EpCAM and Claudin-18 are closely related to the invasiveness of gastric cancer,but abnormally high levels of EpCAM and Claudin-18 are not independent risk factors for the prognosis of gastric cancer patients.

OBJECTIVETo find out the variation of 11 mental element contents in Dendrobium officinale with different germplasms and harvesting ages, the results can provide scientific basis for the quality evaluation and the breeding of D. officinale.

METHOD32 samples with 1-3 ages were collected from cultivated fields of Zhejiang and 11 samples were collected from markets. The 11 mental element contents of samples were determined by ICP-MS or AAS.

RESULTThe average contents of K, Ca, Mg, Mn, Zn, Cr, and Cu were 1,205.23, 766.82, 158.25, 31.06, 4.28, 8.28, and 0.97 mg x kg(-1), the contents of As, Hg, Pb, and Cd were all in limits except Cd content of one sample exceeded the standard limit 0.07 mg x kg(-1); germplasms and physiological ages impacted mental elements contents accumulation significantly.

CONCLUSIONThere were rich essential mental elements in D. officinale. D. officinale from Zhejiang province and medical materials from market were all safe; the breeding of D. officinale can increase the contents of essential mental elements and reduce contents of heavy mental elements; the effect of physiological age on metal elements contents was related to each element's physiological and biochemical function.