BACKGROUND: The nucleus pulposus tissue engineering is regarded as an ideal method in repairing intervertebral disc degeneration. However, the resource of nucleus pulposus cells is limited, so seed cell plays a vital role in nucleus pulposus (NP) tissue engineering. OBJECTIVE: To verify whether adipose-derived stem cells (ADSCs) can differentiate into NP-like cells induced by three-dimensional scaffold. DESIGN, TIME AND SETTING: An in vitro observation based on cell-materials level was performed at the Orthopaedic Research Institute, General Hospital of Beijing Military Area Command of Chinese PLA from July to December 2008. MATERIALS: Three Japanese big-eared rabbits with 3-months old were provided by Vital River Laboratory Animal Technology Co., Ltd., and the medical chitosan was purchased from Golden-shell Biochemical Co., Ltd. METHODS: The ADSCs were isolated from the rabbit adipose and then subcultured with collagenase digestion. The second ADSCs were prepared 50 uL cell suspension (5×10~6 cells), and mixed with 150 uL chitosan-alginate (C/A) and cultured in a12-well plate for 20 minutes in room temperature to obtain C/A gel. The experiment divided into 3 groups, in the control group, 2 mL DMEM containing 10% fetal bovine serum was added; DMEM containing bovine serum albumin, transforming growth factor-1 (TGF), hexadecadrol, vitamin C, bone morphogenetic protein-7,1% ITS was added, and then cultured with 2% O_2 or 20% O_2. All the samples were placed in incubator at a humidified atmosphere of 5% O_2 at 37 V for 21 days. MAIN OUTCOME MEASURES: Histomorphological observation; levels of proteoglycan, as well as type II collagen. RESULTS: The scanning electron microscopy showed that ADSCs grew well on the chitosan-alginate scaffolds, which was positive to oil-O staining and Alcian blue staining. More proteoglycan was produced. With time increasing, there was no significant change in the proteoglycan content and type II collagen in the control group. The content of proteoglycan and collagen type II was higher in the induced group, which had difference to the control group (P < 0.001, P < 0.05). Meantime, the expression of collagen type II and proteoglycan was greater in hypoxia-induced group than that of normoxia-induced group (P<0.05, P<0.01). CONCLUSION: Rabbit ADSCs growing on the scaffolds of chitosan-alginate gel may differentiate towards NP-like cells under certain conditions, especially better under hypoxia state.

Objective:To observe the up-regulation of nuclear Clusterin (nCLU)gene on the biological behaviors of human non-small cell lung cancer cell line A549 .Methods:Sense eukaryotic expression vector of nCLU was constructed by cloning the cDNA of nCLU into pIREShyg3 vector. A549 cells were transfected with pIRES-nCLU and pIREShyg3 vectors by lipofectAMINE~(TM) 2000 mediation, respectively. Stable transfected cells were selected by hygromycin B screening. CCK-8 assay was performed to evaluate the effect of nCLU over-expression on cell proliferation in vitro. The expression level of nGLU protein was examined by Western blotting. Cell cycle distribution was detected by FCM with PI staining. The alteration of migration and metastasis potential of A549 cells before and after nCLU gene transfection were assayed by cell chemotactic migration and invasion test. Results:The proliferation speed of the transfected A549 cell clones stably over-expressing nCLU was slowed down. FCM analyses revealed that the percentage of cells in G_0/G_1 phase dramatically increased from (33.54±2.10)% to (63.31±4.30)%. The cell chemotactic migration and invasion potentials were markedly reduced after nCLU gene transfection (P<0.05). Conclusion:Up-regulation of nCLU can greatly inhibited the proliferation and decreased the migration and invasion capabilities of A549 cells.

Background and purpose:Lung cancer is currently the greatest threat to human life and health of the malignant tumor, clinical examination revealed a solitary pulmonary nodules (SPN), including a signiifcant portion of early stage lung cancer. The research aimed to discuss the diagnosis value of CT guidance Hookwire positioning thoracoscopic surgery for solitary pulmonary nodule. Methods:From Jul. 2011 to Jun. 2013, 310 SPN patients in the Department of Thoracic Surgery of Fudan University Shanghai Cancer Center were collected. Hookwire positioning pins were retained guided by CT scan into the patients’ body. Video assisted thoracic surgery (VATS) pulmonary wedge resection was adopted. According to the result of intraoperative frozen pathology, further treatment method was decided. Positioning accuracy, complications, VATS lung wedge resection surgery successful rate, transfer rate in the chest and SPN pathological classiifcation and other indicators were calculated with statistical methods. Results:Hookwire positioning successful rate was 100%, meanwhile, 2 patients with hemoptysis received symptomatic treatment.Intraoperative Hookwire fell off in 12 patients (3.87%), VATS wedge resection surgery successful rate was 99%, transit thoracotomy was carried out in 3 patients. SPN postoperative histological pathology results:237 cases with primary lung cancer, 73 cases with benign lesions. Conclusion:The method of CT guided Hookwire thoracoscopic surgery after positioning in treatment of SPN has higher diagnostic accuracy rate, reliable curative effect, fewer complications and great value in clinical promotion.

Purpose:To evaluate the effects of intraoperative radiotherapy for adenocarcinoma gastric cardia.Meth- ods:From Jan.1992 to Dec.2000 a trial on intraoperative radiotherapy for AGC was carried out in 38 patients.Whose age was younger than 75 years,at least on semiliquid diet and no contraindication of surgery.6MV or 9MV-E photon was used for the intraoperative radiation therapy to the cardia,lower segment of the esophagus,fundus,lesser curvature and hepato- gastric ligament.Results:Average follow up 65 months.The 5 year survival rates were 39.4%.Conclusions:Intraoperative radiation therapy is able to improve the results of surgery for adenocarcinoma of the gastric cardia.

Objective To investigate the value of CT scans in differential diagnosis of thyroid neoplasm.Methods CT findings ofthyroid neoplasm in 29 cases including 8 cases thyroid adenoma,4 cases nodular goiter and 17 cases thyroid carcinoma identified by operation and pathology were analyzed retrospectively.Results The contour of thyroid adenoma was distinct,a completely enhanced ring in 5 cases and enhanced nodules in 3 cases were showed on contrast-enhanced scan.4 cases of nodular goiter showed homogenous cystic low density the lesions were localized and distinct in contour,the cystic wall could be seen.Thyroid carcinoma appeared unhomogenous density,the edges contour of lesion was not distinct and infiltration on adjacent tissue might be seen.Conclusion CT scan might play an important role in diagnosis and differential diagnosis of thyroid neoplasm.

Objective To evaluate the protective effect of polysaccharide nucleic acid fraction of bacillus Calmette-Guerin (BCG-PSN) against atopic dermatitis (AD) in Nc/Nga mice,and to explore its possible mechanism.Methods Sixteen Nc/Nga mice were classified into normal control group (n =4),low-concentration BCG-PSN group (n=5) and high-concentration BCG-PSN group (n =7) to be subcutaneously injected with sodium chloride physiological solution,BCG-PSN of 0.1 mg/kg and 0.5 mg/kg respectively,at 1,8,15 and 22 days of age.Dinitrochlorobenzene (DNCB) was repeatedly and topically applied to these Nc/Nga mice to induce AD-like lesions at 49 days of age.The preventive effect of BCG-PSN against AD was evaluated by dermatitis scores,scratching frequency,histopathological manifestations and immunological parameters (including IgE,i nterleukin (IL)-4 and-12,and interferon (IFN)-γ).Results Repeated injection of BCG-PSN within 4 weeks after birth significantly decreased the severity of DNCB-induced AD-like lesions,dermatitis scores and scratching behavior in Nc/Nga mice.There was no statistical difference in scratching frequency between the high-and low-concentration BCG-PSN groups.BCG-PSN treatment reduced the plasma level of IgE in Nc/Nga mice in a dose-dependent manner.BCG-PSN at 0.5 mg/kg increased the number of cells secreting IFN-γ in skin lesions of mice.Both doses of BCG-PSN down-regulated IL-4 level,but up-regulated IL-12 level in the culture supernatant of spleen mononuclear cells from mice.Conclusion Early injection of BCG-PSN could protect Nc/Nga mice against dermatitis by promoting the proliferation of IFN-γ-secreting cells,increasing the synthesis of IL-12,and reducing the levels of IL-4 and IgE.

ObjectiveTo observe the effects of hypoxia inducible factor-1 alpha (HIF-1α)-targeting small interfering RNA(siRNA) on the expression of HIF-1α and vascular endothelial growth factor (VEGF) in HaCaT ceils under hypoxic conditions. MethodsHaCaT cells were cultured and divided into four groups, normal control group (without any treatment), hypoxia group (cultured under hypoxic conditions for 24 hours),liposome control group (transfected with liposome followed by hypoxic culture for 24 hours), RNA interference group (transfected with HIF-1α-targeting siRNA/liposome complexes followed by hypoxic culture for 24 hours). Fluorescence real-time quantitative PCR was utilized to determine HIF-1oα and VEGF mRNA expression in HaCaT cells, and Western blot to detect HIF-1α and VEGF protein expression. ResultsNo significant difference was observed in the mRNA expression of HIF-1α between the hypoxia group and normal control group(0.907 ± 0.032 vs. 0.878 ± 0.034, F =1.108, P ＞ 0.05), while the expression levels of VEGF mRNA,HIF-1α and VEGF proteins were significantly higher in the hypoxia group than in the normal control group (0.935 ± 0.032 vs. 0.652 ± 0.053, 0.813 ± 0.047 vs. 0.236 ± 0.014, 0.791 ± 0.030 vs. 0.316 ± 0.013, all P ＜0.05). A significant decline was noted in the mRNA and protein expression levels of VEGF (0.230 ± 0.044 vs.0.978 ± 0.030, 0.213 ± 0.026 vs. 0.817 ± 0.049, both P ＜ 0.05) and HIF-1α(0.497 ± 0.033 vs. 0.806 ±0.040, 0.249 ± 0.028 vs. 0.833 ± 0.052, both P ＜ 0.05) in the RNA interference group than in the liposome control group. ConclusionsHypoxia may enhance the expression of HIF-1α and VEGF in HaCaT cells, and to inhibit the HIF-1α expression may suppress the expression of VEGF in HaCaT cells under hypoxia.

Background and purpose:Gene fusions have been identiifed as recurrent oncogenic events in lung adenocarcinoma. Our purpose are to study the histologic features of anaplastic lymphoma kinase (ALK), c-ros oncogene 1 receptor tyrosine kinase (ROS1) andRETproto-oncogene fusion-positive lung adenocarcinomas and to evaluate the correlation between psammoma bodies and fusion-positive lung adenocarcinomas.Methods:In this study, we performed a comprehensive histologic analysis of 44 fusion-positive (including 15RET, 20ALK and 9ROS1)
lung adenocarcinomas and 111 fusion-negative [including 20 epidermal growth factor receptor (EGFR), 20 Kirsten rat sarcoma viral oncogene (K-ras), 71 pan-negative] lung adenocarcinomas.Results:ALK,RET andROS1 fusion-positive lung adenocarcinomas were more prevalent in solid or acinar predominant adenocarcinoma. Multivariate analysis showed that tumors harboring a fusion gene had significantly higher prevalence of the presence of signet ring cells (P=0.000), micropapillary component (P=0.044), mucinous cribriform pattern (P=0.000) and extracellular mucin (P=0.010). The incidence of psammoma bodies was higher in the lung adenocarcinomas with a gene fusion than in tumors without gene fusions (P=0.000). Psammoma bodies were more likely to be found in tumors with any micropapillary component and/or mucinous cribriform pattern than in tumors lacking a micropapillary component and/or mucinous cribriform pattern (P=0.000).Conclusion:Our data showed that the presence of psammoma bodies, micropapillary component, mucinous cribriform pattern, extracellular mucin or signet ring cells may be either sensitive or speciifc to predict tumors harboring a fusion gene. These distinct morphologic features may be helpful in selecting cases for further accurate molecular testing.

ObjectiveTo evaluate the effects of exercise training on bone histomorphometry of ovariectomized rats.Methods46 SD rats were randomly divided into the normal control group (group A), ovariectomized group (group B) , estrogen supplement group (group C) and exercise training group (group D). The rats of group A had a sham operation, while that of other groups was ovariectomized. 3 months after operation, the rats had been treated for 3 months. The rats of group C were given estradiol, and the rats of group D were trained by exercise. The tibia sections were studied by histomorphometric analysis.ResultsThe values of the trabeclar area, trabeclar area%, trabeclar perimeter and trabeclar number of group D were significantly higher than that of group B. The values of the trabeclar separation, osteoclast number/mm2 and osteoclast number/mm of group D were the least in all groups, which had been ovariectomized. The values of the labeled perimeter % mineral apposition rate, bone formation rate of group D were superior to that of all groups.ConclusionThe exercise training can increase bone mass of ovariectomized rats, which had double effects of promoting bone formation and restraining bone absorption.

Objective To analyze the metastatic rule of common hepatic artery lymph node of thoracic esophageal squamous cell carcinoma,and to investigate the strategies of common hepatic artery lymph node dissection.Methods The clinical data of 682 patients with esophageal squamous cell carcinoma who were admitted to the Cancer Hospital of Fudan University from May 2005 to December 2010 were retrospectively analyzed.The locoregional lymph node metastasis of thoracic esophageal squamous cell carcinoma,relationship between metastatic rates of common hepatic artery lymph node and clinicopathological factors and the postoperative complications were analyzed.The enumeration data were analyzed using the chi-square test.Results A total of 18 277 lymph nodes were dissected (27 lymph nodes per patient).The lymph node metastatic rate was 55.87％ (381/682),and the metastatic lymph node ratio was 7.87％ (1438/18 277).Lymph nodes adjacent to the cardia of stomach,laryngeal nerve,lesser curvature of stomach,cervical esophagus,left gastric artery had a higher metastatic rate,while common hepatic artery lymph node had a lower metastatic rate.All the common hepatic artery lymph node metastasis was accompanied with locoregional metastasis.A total of 1480 common hepatic artery lymph nodes were dissected (2 common hepatic artery lymph nodes per patient).Twenty-four patients had common hepatic artery lymph node metastasis,with the metastatic rate of 3.52％ (24/682) and the lymph node ratio of 2.16％ (32/1480).The common hepatic artery lymph node metastatic rates of upper,middle and lower esophageal squamous cell carcinoma were 2.33％ (1/43),3.76％ (16/425) and 3.27％ (7/217),with no significant difference (x2 =0.295,P ＞ 0.05).The common hepatic artery lymph node metastatic rates of patients in T1,T2 and T3 stages were 2.35％ (2/85),5.46％ (10/183) and 2.90％ (12/414),with no significant difference (x2 =2.850,P ＞ 0.05).The common hepatic artery lymph node metastatic rates of patients with high,moderate and poor differentiated esophageal squamous cell carcinoma were 0(0/63),3.50％ (16/457) and 4.94％ (8/162),with no significant difference (x2=3.259,P ＞ 0.05).The common hepatic artery lymph node metastatic rates of patients with diameter of tumor under 3 cm,3-5 cm and above 5 cm were 2.59％ (6/232),3.02％ (11/364) and 8.14％ (7/86),with significant difference (x2 =6.267,P ＜ 0.05).The common hepatic artery lymph node metastatic rates of patients in N0,N1,N2,N3 stages were 0(0/301),2.53％ (5/198),5.65％ (7/124) and 20.34％ (12/56),with significant difference (x2 =62.368,P ＜ 0.05).The common hepatic artery lymph node metastatic rates of patients in stage Ⅰ,Ⅱ,Ⅲ and Ⅳ were 0(0/62),1.78％ (6/337),5.06％ (13/257) and 19.23％ (5/26),with significant difference (x2=25.959,P ＜0.05).Two hundred and twenty-eight patients had postoperative complications with the complication rate of 33.43％ (228/682).The incidence of anastomotic fistula was the highest,which was 11.58％(79/682).Conclusions The metastatic rates of common hepatic artery lymph node in thoracic esophageal squamous cell carcinoma is the lowest.For patients suffered from esophageal cancer in stage I or the tumor diameter under 5 cm,the dissection of common hepatic lymph node can be ommitted in surgery.