Objective:To elucidate the role of dpc4 gene in the development of squamous cell carcinoma. Methods:DMBA at 5 g/L was locally applied in check pouch 3 times a week for 6,9 and 12 weeks respectively in three groups of golden hamasters to induce carcinoma.The animals were sacrificed after DMBA application.Normal controls were the check pouch samples of golden hamasters without any treatment and negative controls were established by local application of acetone for 12 weeks.DPC4 4 expression was examined by immunohistochemical staining.Results:DPC4 expression was observed in all(22) cases of the normal epithelium samples(100%),20 of the 21 cases of simple hyperplasia(95.2%),20 of the 27 cases of abnormal hyperplasia(74.1%) and 12 of the 25 cases of squomous cell carcinoma(48%).Conclusion:dpc4 may play an important role in the development of oral squomous cell carcinoma.

Objective To observe the intervention efficacy of Tiao Ren Tong Du (regulating the Conception Vessel and unblocking the Governor Vessel) needling in treating perimenopausal insomnia and its effect in improving the symptoms, for providing theoretical evidence for the treatment of perimenopausal insomnia. Method A total of 72 female perimenopausal insomnia patients were randomized into a treatment group (36 cases) and a control group (36 cases). The treatment group was intervened by Tiao Ren Tong Du needling at Baihui (GV20), Shenting (GV24), Yintang (EX-HN3), Zhongwan (CV12), Qihai (CV6), Guanyuan (CV4), Neiguan (PC6), Shenmen (HT7), Sanyinjiao (SP6), and Zusanli (ST36) after standard sterilization, with lifting and thrusting manipulations until needling sensation was obtained. The treatment was given three times a week, successively for 4 weeks. The control group was intervened by conventional public education, without treatment. The Pittsburgh Sleep Quality Index (PSQI) and Kupperman Menopausal Index (MI) were scored before and after the treatment to evaluate the therapeutic efficacy. Result Thirty-three cases in the treatment group finished the treatment and 3 dropped out; while 32 cases in the control group finished the treatment and 4 cases dropped out, and the drop-out rate was 9.7%. The total effective rate was 87.9% in the treatment group versus 40.6% in the control group, and there was a significant difference in comparing the clinical efficacy between the two groups (P<0.01). After the treatment, PSQI and MI sores were significantly changed in thetreatment group (P<0.01,P<0.05). There were no significant changes in PSQI and MI scores in the control group after the treatment (P>0.05). There were significant differences in PSQI and MI scores between the two groups after the treatment (P<0.01,P<0.05). Conclusion Tiao Ren Tong Du needling is effective in treating perimenopausal insomnia, and it can improve perimenopausal syndrome and significantly enhance the patients' quality of life.

BACKGROUND: It is important to study the methods of culturing bone marrow-derived mesenchymal stem cells (MSCs) to obtain a great amount of high purity MSCs for applying ocular tissues constructed by tissue engineering technique to treat eye diseases.OBJECTTVE: To separate and culture in vitro MSCs from bone marrow of the adult rats by density gradient centrifugation combined with adherence culture, and observe the growing characteristics and the possibility of mass multiplication.DESIGN: A completely randomized grouping design/repetitive measuring experiment.SETTING: Pathological laboratory, Zhongshan Ophthalmic Center, Sun Yat-sen University.MATERIALS: Four six-week-old SD rats about 250 g, grade Ⅱ of cleaning, were provided by the Animal Center of Sun Yat-sen University [certificate number: SCSK(Yue)2004/0011], about 250 g each rat and there was no limit to the sex. The main reagents and instruments included low sugar Dulbecco modified Eagle culture medium (DMEM/F12, American Gibco Corporation), trypsin (fetal bovine serum (FBS, Hangzhou Sijiqing Bio-Engineering Material Research Institute), American Gibco Corporation), disodium edetate, lymphocyte separating medium, fibronectin, CD44, CD34, CD31 monoclonal antibodies, two-step-method kit for immunohistochemistry (Beijing Zhong shan Biotechnology Corporation).METHODS: This experiment was conducted at the Key laboratory of Ophthalmology (Sun Yat-sen University), Ministry of ethanol (750 g/L) for 10 minutes. Under aseptic condition, the medullaris cavitas was exposed, the syringe containing application m edium was directly punctured into the femoral cavity, the cells in the medullaris cavitas were washed out with the culture medium containing heparin and taken as the cell suspension. The bone marrow-derived MSCs were separated and purified by density gradient centrifugation combined with adherence culture, and the growing conditions of the wells. When the cells had generally connected with each other, they were fixed with methanol or dimethy ketone in situ for 10 minutes, and then hematoxylin eosin (HE) staining or immunohistochemical staining. Antibodies against fiinoculated to 96-well culture plate by a cell density of 4.25×107/L, with 200 μL every well; then put the culture plate into culture box. Then from the next day to the sixth day, 5 g/L MTT solution was added into two rows (20 μL every well) every day, continuously cultured for 4 hours, then the supernatant was removed, and 200 μL DMSO was added to each well, agitated for 5 minutes, then detected the absorbance (A) values at 570 nm wave length, and a growth curve was drawn.branes were clear and the cell bodies were lucent. Being cultured for 2 days, there appeared adherent cells. 3 days later,most of the adherent cells extended and appeared to be in polygon, star or long-shuttle shapes. 4 days later, the cells showed to be in division growth stage; and about 12 days later, the cell clones were connected to each other, appearing curve of subcultured MSCs was in S shape. Cells began growing fast on the 2nd day after being passaged, and they entered the growth period for 3-4 days followed by saturation period, and then cells stopped growing.CONCLUSION: It is a simple and practical method to separate MSCs from the bone marrow of adult rats by means of density gradient centrifugation and adherence. MSCs can greatly proliferate in vitro and offer seed cells for the application of tissue engineering technique to treat eye diseases.

BACKGROUND: Pterygium is associated with local chronic inflammatory responses and chronic stimulation from external factors such as. sunlight and wind dust. Presently. there are various theories concerning the onset mechanism of pterygium, but these theories are not generally accepted.OBJECTIVE: To investigate histopathological chractefistics of pterygium and analyze the mulfipotent stem cell effects on the onset of pterygium DESIGN: An open experiment.SETTING: Zhongshan Ophthalmic Center of Sun Yat-sen University.MATERIALS: Experiments were performed at the State Key Laboratory of Ophthalmology, Sun Yat-sen University from September 2006 to January 2007. 218 pterygial paraffin specimens following clinical and pathological diagnosis were obtained from Pathology Lab of Zhongshan Ophthalmic Center, Sun Yat-sea University.METHODS: Pterygial specimens harvested from clinical operations received morphology, immunohistochemistry and immunofluorescence under a confocal microscope.MAIN OUTCOME MEASURES: Morphology of ptorygium and expressions of CD34, vimentin (VIM), smooth muscle acfin (SMA), S- 100 in pterygium.RESULTS: Changes in morphology: Fibroplasia and neovascularization were the main changes in pterygium. Fibroplasia wasdiverse in different regions, and two main phenomena were observed. First, the tissues arranged tightly like the scleral fiber.Secondly, in some loose region, some of spindle-shape, polygonal, asteroid fibroblast-like cells, arranging loosely, could be seen only. No apparent collagen fibers were identified between them. Immunohistochemistry were positive for CD34 in some region where the fibroblast actively proliferated, whereas fibrocytes in mature fibrous tissue were negatively stained.Immunohistochemisu'y was positive for VIM in a large fraction of fibrocytes, vascular endothelial cells, vessel wall and perithelial cells. SMA staining was positive in basophilous small blocks, spindle or irregular cell cluster. Of the 218 cases, 56 cases had smooth muscle. S-100 staining demonstrated that neurofilament protein and adipocytes were positive. Of the 218 cases, 44 cases had adipose tissue, Immunofluorescence showed that proliferative active cells were positive and stained green under a confocal microscope.CONCLUSION: The fibrous tissues in pterygium originate from mesenchymal stem cells, and can differentiate into smooth muscle and adipose tissue.

BACKGROUND:Medical stuff have significant impacts on the development of organ donation and transplantation, however, medical students present insufficient knowledge and ambivalent attitude towards organ donation and transplantation. OBJECTIVE:To perform a systemic review on the effectiveness of educational program intervention on medical students’ knowledge and attitudes toward organ donation and transplantation. METHODS:PubMed, WILEY ONLINE LIBRARY, ProQuest Health, Medline, EMbase, CNKI, Wanfang and VIP databases were ful y searched for the published literature of medical students receiving an educational program intervention on organ donation and transplantation. According to the inclusion criteria, randomized control ed trials and quasi-experimental study were evaluated with the standard of Joanna Briggs Institute. The GRADE (Grading of Recommendations Assessment, Development and Evaluation) framework was used to guide the assessment of the methodological quality of the studies. The outcomes were mainly medical students’ knowledge and attitudes toward organ donation and transplantation.
RESULTS AND CONCLUSION:Eight studies from three countries were summarized. The methods were employed in two studies of randomized control ed trials and quasi-experiment received a high quality rating, five studies received middle quality rating and one study received low quality rating. Educational program increased the knowledge in six studies, and changed the attitudes in six studies. Educational program intervention is a significant strategy to increase medical student’s knowledge about organ donation and transplantation. The majority of involved study are
quasi-experiment study, so more high-quality randomized control ed studies are needed.

Objective To observe the anti-tumor effect and mechanism of curcumin on triple-negative breast cancer mouse model.Methods Human breast cancer cells MDA-MB-231 were inoculated to the chest subcutaneous fat pad of mice(about 1 million cells in 0.1 mL cell suspension).At day 2 after tumor inoculation,nodules were seen at inoculation site (tumor formation rate being 100％).Forty successfully-modeled mice were randomly divided into low-,middle-,and high-dose curcumin groups (10,20,40 mg/kg) and model control group,10 mice in each group.The medication lasted for 30 continuous days.After medication,the blood was taken out from the orbital venous plexus,and the serum was separated for the detection of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) contents.Spleen was separated for the calculation of organ coefficient,tumor mass was weighed,and the expression levels of Bcl-2 and caspase-3 were detected by Western blotting method.Results Compared with the model control group,caspase-3 expression level was significantly increased in the three curcumin groups(P ＜ 0.05 or P ＜ 0.01),but the contents of TNF-α and IL-6,tumor weight,spleen index and Bcl-2 expression were significantly decreased(P ＜ 0.05 or P ＜ 0.01).Conclusion Curcumin has inhibitory effect on triple-negative breast cancer mouse model through promoting the apoptosis of breast cancer cells.

Objective To investigate the changes between type 2 diabetes mellitus(T2DM) with peripheral vessel disease and osteoporosis. Methods A total of 100 cases T2DM patients were divided into non-peripheral vessel disease (A group) and peripheral vessel disease (B group). B-mode ultrasonography to measure the intima-media thickness(IMT) of carotid artery,dual energy X-ray absorptiometry (DEXA) to measured bone mineral density(BMD) and VS-1000 arteriosclerosis diagnometer to measure brachi-ankle pulse wave velocity (baPWV) were used in 100 T2DM patients. The clinical data and BMD were compared between two groups. Results Compared with normal group,the BMD in T2DM patients significantly decreased.Compared with T2DM patients without peripheral vessel disease,the BMD decreased in T2DM patients with peripheral vessel disease. Conclusions Atherosclerosis in T2DM with peripheral vessel disease is probably related to osteoporosis, and the relationship between them is worth further investigation.

Objective To investigate the expression of cyclooxygenase-2(COX-2)in breast cancer tissues,as well as the relationship between COX-2 and the clinicopathological features.Method The expression of COX-2 was detected in 60 cases of breast cancer tissues and 20 cases of breast normal tissues by using immunohistochemistry,and combined with clinicopathological information for analysis.Results The COX-2 expression rate was 65.0%(39/60)in breast cancer tissues and 10.0%(2/20)in breast normal tissues respectively.There was statistic difference between the two(P<0.01).The over expression of COX-2 was significantly correlated with TNM stages,lymphatic metastasis and the expression of epidermal growuth factor receptor-2(C-erbB-2)(P<0.05 or<0.01).Conclusion The expression of COX-2 in breast cancer tissues is significantly higher,which might play a fairly important role in tumorigenesis and progression of breast cancer.

Objective:To explore the relationship between apoptotic protease activating factor 1(APAF1) gene and oral squamous cell carcinoma (OSCC). Methods:The mRNA expression of APAF1 gene were detected with semiquantitative RT-PCR method in 18 cases of normal oral mucous membrane and 32 cases of OSCC tissues. Results:The expression of APAF1 gene was significantly decreased in OSCC tissues comparing with the normal oral mucous membrane(P

Objective To investigate the effects of inhaled low dose nitric oxide(NO)on lung ischemia-reperfusion injury during flush and delayed 10 min after reperfusion.Methods Sixty health a- dult male Sprague-Dawley rats were randomly allocated to the control and the NO group.Before the donor lung was harvested,the right hilus was clipped for 5 min(clipping test),then blood sample was collected from carotid artery for arterial blood gas analysis as baseline.Lung transplantation was per- formed in a“cuff-like”vessel anastomosis technique.Dynamic compliance(Cdyn)and resistance of airway(Raw)were monitored before operation(baseline)and after 2-h reperfusion.The graft's gas exchange and oxygenation were assessed by“clipping test”after 2-h reperfusion.The lung graft was harvested for measuring wet/dry weight ratio(W/D),the activity of myeloperoxidase(MPO)and in- ducible nitric oxide synthase(iNOS),the content of malonyldialdehyde(MDA),and the expression of iNOS gene and protein.Results After 2-h reperfusion,compared to the control group,PaO_2/FiO_2, OI,and Qs/Qt were improved significantly in the NO group(277?91 vs.157?47,P＜0.01;2.67?0.89 vs.4.72?1.48,P＜0.01;21.1?4.57 vs.27.1?2.37,P＜0.01,respectively).The activi- ties of MPO were significantly reduced in NO group(1.80?0.46 vs 3.08?0.65 U/g tissue,P＜0.01).The content of MDA in the lung tissue of NO group was significantly higher than that of the control group(34.8?7.9 vs.20.0?11.2 nmol/mg protein,P＜0.05).Inflammatory cell infiltration was also significantly reduced(P＜0.05).The expression of iNOS gene and protein in the lung tissue of NO group was significantly lower than that of the control group.The activities of iNOS were also significantly reduced in NO group(10.6?10.2 vs 97.8?82.2 nmol?g~(-1)?min~(-1),P＜0.05).The im- munohistochemical positive staining of iNOS was localized in the alveolar epithelial cells and the in- flammatory cells infiltrated in the alveolar spaces and mesenchymal tissue.But there were no signifi- cant differences between two groups in Cdyn,Raw and W/D ratio.Conclusion Inhaled low dose NO might mitigate the intrapulmonary shunt,prevent neutrophil sequestration,inhibit the expression of iNOS gene and protein in isograft,thereby ameliorate ischemia-reperfusion injury and improve the ox- ygenation of the graft.