Objective To observe the change endoplasmic reticulum stress in renal injury of fluorosis rats,and explore the effect of endoplasmic reticulum stree in the mechanism of renal injury of fluorosis.Methods 48 Wistar rats were divided into 4 groups:control,fluoride-treated,low calcium,and low calcium plus fluoride-treated.The rats in fluoride-treated and low calcium plus fluoride-treated groups were treated with sodium fluoride (NaF,221 mg?L-1) through drinking water for 3 months.The pathological slice of rat kidney was made and the morphological changes were observed under optical microscope.The transcription levels of Grp78,Xbp1,CHOP and PDI in the kidney tissues were determined by reverse transcription-polymerase chain reaction (RT-PCR).Results Edema and vacuolar degeneration of the proximal tubule and distal tubule cells in the kidney tissues in fluoride-treated group were observed;and cell edema,vacuolar degeneration,scattered necrosis,minor regeneration repair and interstitial hyperemia in the kidney tissues in low calcium plus fluoride-treated group were found.The mRNA expressions of Xbp1 in the kidney tissues in fluoride-treated and low calcium plus fluoride-treated groups were markedly higher than that in the responding control groups(P

BACKGROUND: In tissue engineering, three-dimensional biodegradable scaffolds are generally used as a basic structure for cell anchorage, proliferation. Currently, no perfect scaffold is available. OBJECTIVE: To observe the growth of rabbit bone marrow stromal cells (BMSCs) cultured in different-intensity three-dimensional fibrin glue in vitro, and to discuss the feasibility of fibrin glue used as a scaffold material of bone tissue engineering. DESIGN, TIME AND SETTING: The single sample observational study was performed at the China-Japan Union Hospital of Jilin University and School of Mechanical Engineering of Tianjin University of Technology from September 2007 to January 2008. MATERIALS: Fibrinogen and thrombin were mixed at various proportions, and prepared into different intensity fibrin glue. A month-old male New Zealand white rabbits, weighing 0.25 kg was used in this study. METHODS: Rabbit BMSCs were cultured and serial subcultivation in a CO2 incubator. And then the amplified BMSCs were collected and continue to be cultured in different intensity fibrin glue for 4 weeks. MAIN OUTCOME MEASURES：Observation of growing BMSCs is performed using the phase contrast microscope. The activity of BMSCs in fibrin glue at different stages was observed using hematoxylin-eosin staining. The ultrastructural changes of BMSCs were observed which had been cultivated in fibrin glue for 4 weeks. RESULTS: After growing in fibrin glue for 4 weeks, BMSCs showed strongly active status in low intensity fibrin glue and growing slowly or dying in high intensity fibrin glue. Under the electron microscope, BMSCs following 4 weeks culture in fibrin glue (proportation of fibrinogen and thrombin was 4:1) were found, with visible cellular organs, and BMSCs had good activities. CONCLUSION: BMSCs can spread and proliferate quickly in low intensity fibrin glue. The optimal proportion of fibrinogen and thrombin is 4: 1.

BACKGROUND: A superior composite scaffold hopes be constructed to resolve adhesion between seed cell and scaffoldmaterial.OBJECTIVE: To construct composite scaffolds with fibrin glue and xenogeneic inorganic bone and to explore the three-dimensional culture of rabbit marrow stromal cells (MSCs).DESIGN, TIME AND SETTING: Observational experiment was performed at the Department of Orthopedics of China-Japan Union Hospital and the Department of Mechanical Engineering, College of Mechanical Engineering of Jilin University from November 2007 to March 2008.MATERIALS: Fibrin glue was made by a certain ratio of fibrinogen and thrombin; bovine cancellous bone following defatting and deproteinization was mixed with fibrin glue to establish composite scaffold.METHODS: Rabbit MSCs were cultured in v#ro and transferred, and the MSCs were collected for three-dimensional culture withcombined scaffolds made of xenogeneic inorganic bone and fibrin glue.MAIN OUTCOME MEASURES: The growth and proliferation of MSCs were examined by phase-contrast microscope andtransmission electron microscopy.RESULTS: Phase contrast microscope showed that the MSCs could spread evenly in the combined scaffolds. After cultured 4 weeks, the MSCs formed into densely three-dimensional net. It could be observed under the transmission electron microscopethat there were micro-protrusions in local stromal cells at 4 weeks after culture, and the mitochondrion as well as ribosomes wasobserved in the cytoplasm with rough endoplasmic reticulum.CONCLUSION: The MSCs cultured in the combination of fibrin glue and xenogeneic inorganic bone scaffolds show a betteractivity, and they can proliferate rapidly.

Objective To examine the expressions of IKKε protein in the specimens and cells of epithelial ovarian cancer and investigate the effect of IKKε inhibitor on cell proliferation and apoptosis. Methods (1) A total of 118 cases of patients with the median age of 59 who have accepted surgical treatment due to ovarian cancer in the First Affiliated Hospital of Dalian Medical University from January 2006 to April 2013 were selected. Twenty cases of patients with the median age of 55 who have accepted hysterectomy and salpingo-oophorectomy due to uterine leiomyoma during the same period were selected as the control. The expressions of IKKε protein were detected by immunohistochemistry in normal ovarian tissues and epithelial ovarian cancer specimens,and the relationship between the expressions of IKKε and the clinical features of patients was analyzed. IKKε protein was determined by western blot in various ovarian cancer cells, including SKOV3, OV2008, C13, A2780S, A2780CP, OV4, OV5, OV8, and CAOV3 treated with or without IKKε inhibitor. The cellular proliferation and apoptosis of ovarian cancer cells after 48 hours treatment of IKKε inhibitor were analyzed by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry, respectively. Results (1) The immunohistochemical results showed that IKKε was highly expressed in epithelial ovarian cancer specimens with the expression rate 66.1% (78/118), compared with normal ovarian tissue with the expression rate 35.0% (7/20), which exhibited statistically significant difference (χ2=6.993, P=0.008). The expression of IKKε protein was correlated with International Federation of Gynecology and Obstetrics (FIGO) stage, histological grade, the level of CA125 in preoperative serum and distribution of the tumor (P<0.05), but no correlation with age, histological type, the incidence pattern, and tumor size (all P>0.05). (2) IKKε was widely overexpressed in different levels in SKOV3, OV2008, C13, A2780S, A2780CP, OV4, OV5, OV8, and CAOV3 cells, and the expression of IKKε decreased as the increase of the concentration of IKKε inhibitor (0.1 and 0.5 μmol/L) in OV2008, C13, A2780S, and A2780CP cells after 48 hours treatment. Different concentrations of IKKε inhibitor (0.05, 0.1, 0.5, 1, 5, 10, and 25 μmol/L) significantly inhibited the proliferation of OV2008, C13, A2780S, A2780CP, and SKOV3 cells in a concentration-dependent manner (P<0.05), and the half maximal inhibitory concentration (IC50) was 0.43, 0.86, 0.10, 0.19, and 0.24 μmol/L, respectively. The cell apoptotic rate of OV2008, C13, A2780S, A2780CP, and SKOV3 cells was significantly increased after 48 hours treatment of IKKεinhibitor with the concentration of 0.1 and 0.5 μmol/L (P<0.05). Conclusions The IKKε protein in epithelial ovarian cancer specimens and cells is overexpressed. IKKε inhibitor could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner. Together, the result indicated that IKKε may be a candidate target for the treatment of ovarian cancer in future.

BACKGROUND: Osteolysis has always occurred in pelvis. Percutaneous injection of bone cement stabilized bone fracture, relieved pain or even treated tumor. However, leakage of bone cement might cause severe complications. OBJECTIVE: To explore the therapeutic effect of peroutaneous injection of bone cement on treating osteolysis pelvic disease in 9 cases by the CT guidance. METHODS: By the CT guidance, needing degree was determined firstly. Focal size and scanning layers were used to calculate focal volume and estimate injected dose of bone cement. Three-dimensional targeting device was used to introduce the puncturation. The bone cement which was 0.2-0.5 mL less than the calculated volume was injected into osteolysis site. The accuracy, injected dose, clinical efficacy, and complications were investigated. RESULTS AND CONCLUSION: The following-up ranged from 5 months to 4 years, with mean duration of 1.5 years. At 1-48 hours after operation, symptoms were recovered, including complete recovery (n=6), partial recovery (n=2), and light recovery (n=1). Leakage of bone cement was not detected out around focal region. This suggested that percutaneous injection of bone cement into the erosion site is an effective method to treat pelvic osteolysis disease, characterizing by security, effective, and less invasive.

This study investigated the effect of arctigenin (Arc) on the cell activation, cytokines expression, proliferation, and cell-cycle distribution of mouse T lymphocytes. Mouse lymphocytes were prepared from lymph node and treated with Phorbol-12-myristate-13-acetate (PMA)/Ionimycin (Ion) and/or Arc. CD69, CD25, cytokines, proliferation and cell cycle were assayed by flow cytometry. The results showed that, at concentrations of less than 1.00 micromol x L(-1), Arc expressed non-obvious cell damage to cultured lymphocytes, however, it could significantly down-regulate the expression of CD69 and CD25, as well as TNF-alpha, IFN-gamma, IL-2, IL-4, IL-6 and IL-10 on PMA/Ion stimulated lymphocytes. At the same time, Arc could also inhibit the proliferation of PMA/Ion-activated lymphocytes and exhibited lymphocyte G 0/G1 phase cycle arrest. These results suggest that Arc possesses significant anti-inflammatory effects that may be mediated through the regulation of cell activation, cytokines expression and cell proliferation.

Objective To detect and explore the expression and clinical significance of dual specificity tyrosine phosphorylation regulated kinase1b (Dyrk1b) in the specimens and cells of cervical lesions. Methods (1)All the data were collected from 75 patients with cervical cancer and 52 cases with squamous intraepithelial lesion(SIL)admitted in the First Affiliated Hospital of Dalian Medical College during Jan. 2011 to Dec. 2013 and confirmed by pathological examination, included 60 cases of stageⅠand 15 cases of stageⅡ, 12 cases with low-grade squamous intraepithelial lesion(LSIL)and 40 cases with high-grade squamous intraepithelial lesion(HSIL). While, 28 cases with chronic cervicitis were chosen as the control group. The protein expression of Dyrk1b was detected by immunohistochemistry among the four groups.(2)The expression of Dyrk1b in HeLa and SiHa cells were detected by western blot method and the expression of Dyrk1b protein were also detected after treatment of AZ191 (5, 10 μmol/L) for 48 hours in HeLa and SiHa cells.(3)The cellular survival and proliferation of HeLa and SiHa cells treated by different concentrations of AZ191(2.5, 5, 10, 25, 50, 100 μmol/L)for 48 hours were detected by methyl thiazolyl tetrazolium (MTT) assay.(4)The rate of apoptosis of HeLa and SiHa cells was detected by flowcytometry after treatment of AZ191 (5, 10μmol/L) for 48 hours. Results (1)The positive rates of Dyrk1b protein in chronic cervicitis, LSIL, HSIL and cervical squamous cancer by immunohistochemistry were 11%(3/28), 1/12, 42%(17/40)and 71%(53/75), respectively. The expression of Dyrk1b in cervical squamous cancer and HISL were higher than those in LSIL and chronic cervicitis (P<0.01), there were significant difference between cervical squamous cancer and HSIL, or between HSIL and LSIL(all P<0.05), while there were not significant difference between LSIL and chronic cervicitis(P>0.05). Expression of Dyrk1b was correlated with stromal invasion depth of cervical cancer (P<0.05), but not with age, clinical stage, lymph node metastasis, and serum squamous cell carcinom antigen(SCC-Ag)levels (all P>0.05). (2) Dyrk1b protein was expressed in different levels in HeLa and SiHa cells, and the expression of Dyrk1b was decreased gradually as the increased of the concentration of AZ191 in both HeLa and SiHa cells by treatment of AZ191 for 48 hours. (3) Different concentration of AZ191 treated on cervical cancer cells could inhibit the cellular proliferation and induce cell apoptosis in a concentration-dependent manner(P<0.01), concomitant to the decreased cell survival rate. The apoptosis rate of HeLa and SiHa were increased significantly after 10μmol/L AZ191-treatment for 48 hours, but no any difference induced by 5 μmol/L AZ191-treatment compared to control group. Also,there was no any difference between Hela and SiHa cells in either inhibitory effect or apoptosis rate induced by AZ191. Conclusions Dyrk1b is over-expressed in either specimens or cells of cervical cancer. The expression of Dyrk1b protein in cervical lesions is increased as the progression of disease. Dyrk1b inhibitor AZ191 could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner in cervical cancer cells.

Suppressor APC domain containing 2 (SAPCD2) gene may affect tumor proliferation, invasion, migration and apoptosis, and participate in tumor-related signal pathway transduction, suggesting that SAPCD2 may be a biomarker of tumor and a potential target for treatment. Therefore, SAPCD2 has been widely concerned by researchers. This article reviews the role and related mechanisms of SAPCD2 in tumors.

Objective To investigate the prevailing situation concerning psychological crisis intervention abilities of college staff,and provide countermeasures and suggestions for further strengthening the psychological crisis intervention abilities of college counselors.Methods A"Questionnaire on the Structure of Psychological Crisis Intervention Ability of College Counselors,"was conducted among 120 college counselors and 240 college students from eight colleges in Shenyang.A total of 92 counselors and 199 college students'responses were collected,with an effective recovery rate of 80.83%.Results There were statistical differences between male counselors and female counselors in personality trait dimension(t=-2.156,P＜0.05).There were also statistical differences between counselors who had dealt with students'psychological crises and those who had not in terms of the knowledge(t=-2.786,P＜0.01)and ability dimen-sions(t=-2.151,P＜0.05).Statistical differences were also observed in the evaluation of counselors'crisis intervention ability between students who had experienced psychological crisis events and those who had not.Lastly,there were statistical differences in the empathy(t=-3.176,P＜0.01)and personality trait dimensions(t=-2.424,P＜0.05)between counselors'self-evaluation and students'evaluation.Conclusion At present,college counselors have improved intervention capacity in dealing with students'psychological crises,but it is still necessary to strengthen the preventive role in psychological crisis intervention.Universities need to strengthen speciali-zed training on psychological crisis intervention and provide resources and support for counselors.

Objective To detect phosphorylated-extracellular signal-regulated kinase (p-ERK1/2) protein expression in epithelial ovarian cancer and cell lines,and to examine the effects of mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor AZD6244 on cell proliferation,apoptosis as well as cell cycle of ovarian cancer cells.To explore the function and significance of MAPK/extracellular signal-regulated kinase (ERK) signaling pathway in the development of ovarian cancer.Methods (1) A total of 104 cases of patients with ovarian cancer who accepted the treatment of gynecological surgery and being confirmed by pathological examination in First Affiliated Hospital,Dalian Medical University from January 2004 to December 2013 were selected.The expressions of p-ERK1/2 protein were detected by immunohistochemistry in ovarian cancer specimens,and the relationship between the expressions of p-ERK1/2 and the clinical features of patients was analyzed.(2) p-ERK1/2 and other related proteins were determined by western blot in various ovarian cancer cells,including SKOV3,OV2008,C13,A2780S,A2780CP,OVCAR4,OVCAR5,OVCAR8 and CAOV3 treated with or without MEK inhibitor.The cellular proliferation,apoptosis and cell cycle of ovarian cancer cells after treatment with MEK inhibitor were analyzed by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry,respectively.Results (1) The immunohistochemical method showed that p-ERK1/2 between low grade serous carcinoma and clear cell carcinoma were not significantly higher expressed (P＞0.05).However,a lower level of the p-ERK1/2 expression were observed among high grade serous carcinoma,mucinous carcinoma and endometrioid carcinoma (all P＜0.05).There was no significant correlation between the protein expression of p-ERK1/2 and patients' age,pathological stage of surgery,and preoperative serum CA125 level (P＞0.05).(2) Western blot showed that the protein p-ERK1/2 was widely expressed in various ovarian cancer cell lines such as SKOV3,OV2008,C13,A2780S,A2780CP,OVCAR4,OVCAR5,OVCAR8 and CAOV3.After treatment with AZD6244 (5,10 μmol/L),the level of p-ERK 1/2 in OVCAR5 and OVCAR8 decreased significantly in dose-dependent manner.Additionally,we found a reduction of the expression level of cyclin D 1,caspase-3 and appeared cleaved poly adenosine diphosphate ribose polymerase (PARP) in OVCAR5 and OVCAR8,compared with control groups.MTT assays showed that OVCAR5,OVCAR8 and A2780S were differently inhibited in the dose-dependent manner after being treated with different concentrations of AZD6244 (0,2.5,5,10,25,50 and 100 μmol/L,all P＜0.05).Further tested by flow cytometry,the results showed that AZD6244 (5,10 μmol/L) was able to induce the apoptosis of OVCAR5,OVCAR8 and A2780S,as well as G0/G1 phase arrest,both in a dose-dependent manner (P＜0.05).Conclusions As the main active and functional unit of MAPK/ERK signaling pathway,p-ERK1/2 protein is expressed in both the tissues and various ovarian cancer cell lines.AZD6244 could down-regulated the expression of p-ERK1/2 in ovarian cancer cells,accompanied by the decreased proliferation and increased cell apoptosis of ovarian cancer cells.In conclusion,MAPK/ERK signaling pathway might play a role in the development and progression of ovarian cancer,and may be provide a novel option for molecular targeted therapies of the disease.