The incidence of cholangiocarcinoma is rising, poor prognosis due to lack of viable treatment.Recent investigations into the underlying molecular mechanisms of cholangiocarcinogenesis and tumor growth have contributed greatly to our understanding of this disease, which will lead to the identification of therapeutic targets for this devastating cancer. The purpose of the following review is to address what has been learned over the past decade concerning the molecular basis of cholangiocarcinogenesis and factors regulating cholangiocarcinoma growth. Through a better understanding of these mechanisms, more specific diagnostic, therapeutic, and preventative strategies may be developed and hopefully improve the outcome of this devastating disease.

Objective To establish a purified model of rat retinal ganglion cells (RGCs) cultured by serum-free medium,and provide a good cell model to investigate the damage of RGCs in glaucoma,retinal ischemia,and degenerative retinopathy. Methods Two monoclonal antibodies,anti-rat SIRP (OX-41) against rat macrophage and antibody against rat Thy-1 (OX-7),were used to purify and characterize RGCs from 1-3-day old Sprague-Dawley (SD) rats by means of two-step filtration.Purified RGCs were cultured in serum-free neurobasal medium containing B27 and ciliary neurotrophic factor (CNTF) meeting the neuronal cell's special requirements.Photomicrographs illustration,immunfluorescence staining of Thy-1,calcein-acetoxymethyl ester (calcein-AM) fluorescence images were used to observe and identify cultured retinal cells and purified RGCs. Results Among the primary cultured rat retinal cells,91% were retinal neurons.Protuberances of RGCs were seen after cultured for 24 hours.At the4th to 8th day,many cells had uniform configuration,large body,and long protuberances. At the 14th day,over 60% cells maintained viability.Immunoflurescence staining of Thy-1 showed the purity of RGCs was about 90%. The results of calcein-AM staining,which stained the living cells only,showed large cell body of RGCs and most of RGCs had a protuberance whose length was twice longer than the diameter of the cells. Conclusion RGCs cultured by serum-free medium has uniform size,good configuration,and high purity,which is adapt to the research of damage of RGCs caused by various factors and to evaluate the protective effects of neuroprotective agents.

Objective To establish the ranges of normal reference values of fetal brainstem-vermis angle (BVA)and brainstem-tentorium angle (BTA)in normal fetuses at different gestational weeks,and to evaluate their clinical values in the diagnosis of mega cisterna magna.Methods Totally 320 normal singleton fetuses between 1 8 and 40 weeks of gestation were enrolled in this study.Three-dimensional ultrasound (3DUS)with volume contrast imaging (VCI)was used to measure their BVA and BTA,the correlations of BVA and BTA with gestational age were analyzed.The BVA and BTA of 24 fetuses with mega cisterna magna were also measured and were compared with those of the normal fetuses.Results In these normal fetuses,the BVA and BTA showed no correlations with gestational age (r=-0.05,0.04,both P >0.05);the ranges of normal reference values of BVA and BTA were 2.23°-19.30°and 18.53°-48.13°, respectively.The BVA and BTA of 24 fetuses with mega cisterna magna were 4.23°-17.42°and 20.41°-40.89°,respectively,which were not significantly different from those of normal fetuses (t =-0.55,0.92, both P >0.05).Conclusions 3DUS with VCI is feasible for measuring the BVA and BTA of fetuses,and the measured values can be used as the reference standards for BVA and BTA measurement in China.

Objective To observe the psychological stress reactions of 238 patients injured in earthquake in our hospital and explore their interventions to such reaction. Methods A total of 238 patients wounded in earthquake were recruited. Along with the active treatment of primary physical trauma, wide range of preventative psychological interventions for crisis was carried out individually or in group. The psychological emergency measures included communication and exchange of ideas, mental health education, and psychological support and a relaxing environment to the patients. All the patients were evaluated by using depression and anxiety assessment scales before and after the intervention. Results Various psychological stress responses were found in 238 patients after injury, in which sleep disorders, memory decline, tension and fear, wabble and emotional instability ranked the top 5. Some patients' psychological status recovered after one-week's hospitalization, while most recovered after two-week's hospitalization. Three patients were transferred to other professional psychiatric hospitals for professional psychological treatment and intervention because of serious psychological obstacle. Conclusion The wounded caused by earthquake manifested various psychological stress reactions. Immediate psychological intervention can effectively help the patients to recover psychologically, promote their physical rehabilitation, and prevent posttraumatic stress disorder.

Objective To investigate the role of natural killer cells (NK cells) in patients with chronic hepatitis B (CHB). Methods Blood samples and liver biopsies from 108 CHB patients including 43 mild, 28 moderate and 37 severe CHB patients were collected. The levels of interferon (IFN) gamma in liver tissues and peripheral blood were detected by intracellular cytokine staining.The data were analyzed by t test. Results The levels of IFN gamma produced by NK cells in liver tissues and peripheral blood of patients with severe CHB were (8. 12±4. 26) % and (4. 59±2. 95) % ,respectively, which were significantly higher than those in patients with mild CHB [(3. 53±1. 56)%and (2. 26±0.61)%, respectively; t=3. 80 and t = 4. 30, respectively; both P<0. 05] and those in patients with moderate CHB [(4. 59±1. 98)% and (2. 48±1. 05)%, respectively;t=2. 62 and t =3. 28, respectively; both P<0. 05]. While there were no differences of IFN gamma produced by NK cells both in liver tissues and peripheral blood between mild and moderate CHB patients (t=0. 99 and t=1. 27, respectively; both P>0. 05). The levels of IFN gamma produced by NK cells in liver tissues and blood samples in severe phase of severe CHB were (10. 02 ± 4. 15)% and (6. 78 + 2. 91)%, respectively, those in recovery phase were (6. 13±2. 01)% and (3. 13± 1. 52)%, respectively (t = 1. 89 and t=1. 74, respectively; both P<0. 05]. Conclusion NK cells may play a pivotal role in the pathogenesis of severe CHB.

Objective To explore the expression of KAI1/CD82,E-cadherin and β-catenin in endometrial carcinoma,and to investigate their correlations to clinicopathological parameters of endometrial carcinoma. Methods The expressions of KAI1/CD82,E-cadherin and β-catenin in 76 specimens of endometrial carcinoma,15 specimens of atypical endometrial hyperplasia and 20 specimens of proliferative endometrium were examined by immunohistochemical envision technique.Their correlations to clinicopathological parameters of endometrial carcinoma were statistically analyzed. Results Compare to normal proliferative phase endometrium and atypical endometrial hyperplasia,the expression of KAI1/CD82 in endometrial carcinoma was significantly decreased(P ＜0.01),the abnormal expression of E-cadherin and β-catenin in endometrial carcinoma were significantly higher(all P <0.01).In endometrial carcinoma,the expression of KAI1/CD82 was negative correlated with histological grade and depth of myometrial invasion(P <0.01,P <0.05); The abnormal expression of the E-cadherin is related to histological grade and type(P <0.01,P<0.05); The abnormal expression of β-catenin was positively correlated with histological grade and FIGO stage(P ＜0.01 ,P <0.05).The down-regulation expression of KAI1/CD82 was closely associated with the abnormal expression of E-cadherin and beta-catenin in endometrial carcinoma(P <0.01,P <0.05). Conclusion The down-regulation of KAI1/CD82 and the aberrant expression of E-cadherin and β-catenin could be involved in the development of endometrial carcinoma.The loss or reduced expression of KAI1/CD82 was closely associated with the abnormal expression of E-cadherin and β-catenin in endometrial carcinoma.

Objective The aim of this study was to examine the association between plasma Pentraxin 3(PTX3) levels with acute ischemic stroke and carotid artery atherosclerosis.Methods We enrolled 103 patients with acute ischemic stroke (AIS)and 83 control subjects.The levels of plasma PTX3 were measured by using Enzyme-linked immunosorbent assay (ELISA) at admission and after 7 days treatment in the AIS group.The carotid artery plaques in the AIS group were detected by using Color Doppler Ultrasound.Patients with AIS were divided into two groups according to the stability of carotid artery atherosclerosis plaques.The association between plasma PTX3 levels with acute ischemic stroke and the stability of carotid artery atherosclerosis plaques was examined.Results ① The plasma PTX3 levels were significantly higher in the AIS group than in the control group (P＜0.05).After 7 days standard treatment,the plasma PTX3 levels in AIS group were significantly decreased (P＜0.05),but still higher than those in control group (P＜0.05).Multivariable logistic regression suggested that the plasma PTX3 levels had a close relationship with AIS (OR=15.043,95％CI:3.46～65.45,P＜0.001).② In the AIS group,the plasma PTX3 levels before and after treatment were significantly higher in unstable plaque group than in no plaque and stable plaque group(P＜0.05).Conclusion The plasma PTX3 levels are higher in patients with acute ischemic stroke.The plasma PTX3 levels are significantly higher in AIS group with unstable plaque.The plasma PTX3 levels are closely associated with acute ischemic stroke.

Objective To investigate the mecbanism of safflor injection treating myocardial contusion following thoracic impact.Methods Thirty healthy Japan big-ear rabbits were randomly assigned to the myocardial contusion control group (control group,n =15) and safflor injection group (safflor group,n =15).The rabbits with severe myocardial contusion were induced by BIM-Ⅱ Horizontal Bioimpact Machine.Blood samples were taken from common carotid artery of both groups to measure the levels of plasma thromboxance A2 (TXA2) and prostaglandin I2 ( PGI2 ) at 10 minutes before impact and at t0 minutes,30 minutes,1,3,6,12 and 24 hours after impact.Rabbits were dissected to observe the gross and micropathological changes of the hern at 24 hours after impact.Results The control group showed a distinct falling of plasma PGI2 concentration and a significant increase of plasma TXA2 concentration in the early period after severe myocardial contusion.The morphological observation revealed severe myocardial tissue injury as well.However,the safflor group showed insignificant variation pertaining to the reduction of plasma PGI2 concentration and increase of plasma TXA2 concentration,but obvious alleviation of myocardial tissue injury,as compared with the control group.Meanwhile,the ratio of TXB2 to 6- Ketoprostaglandin Fla (6-Keto-PGFla) in the safflor group was lower than that of the control group (P ＜0.05).Conclusion Safflor injection has therapeutic effect on myocardial contusion by elevating the concentration of plasma PGI2 and decreasing the concentration of plasma TXA2 in the early period after severe myocardial contusion.

Objective To study the expression of programmed death 1(PD-1) on the peripheral T lymphocytes and the serum cytokine levels in different phases of hepatitis B virus (HBV) infection (immune tolerance,immune clearance and non-replicating phases).Methods A total of 105 HBV infected patients in different phases of infection were enrolled in Wuxi No.5 People's Hospital from Apr to Sep 2011,and divided into three groups:the immune tolerance group (35 cases),the immune clearance group (35 cases) and the non-replicating group (35 cases).Meanwhile,20 healthy people were enrolled as controls.The peripheral blood was collected and PD-1 expressions on the surface of T lymphocytes were assessed by flow cytometry.The cytokine levels in different groups were analyzed by enzyme-linked immunosorbent assay (ELISA).The data were analyzed by t test and Pearson correlation analysis.Results The PD-1 expression on surface of CD8+ T lymphocytes of HBV infected patients was (12.35± 3.48)％,which was significantly higher than healthy control group [(4.65±1.21) ％] (t=9.76,P＜0.01).The PD-1 expression on CD4+ T lymphocytes was (4.95±2.87) ％,which was not significantly different from the healthy controls [(4.08±2.14) ％] (t =1.29,P＞0.05).The PD-1 expressions on CD8+ T lymphocytes of patients in the immune tolerance group,the immune clearance group and the non-replicating group were (15.87±3.18)％,(7.69±3.64)％and (10.12±2.84) ％,respectively; that in the immune tolerance group was higher than the immune clearance group (t=10.01,P＜0.01) and that in healthy control group was lower than those in the other 3 groups (t=15.12,3.61 and 8.17,respectively; all P＜0.01).The PD-1 expression on the surface of CD8+ T lymphocytes in chronic HBV infected patients was positively correlated with the serum interleukin-10 (IL-10) level (γ=0.377,P＜0.05),while that was negatively correlated with interferon-γ (IFN-γ) level (r=-0.620,P＜0.05).Conclusions PD-1 is up-regulated on the surface of CD8+ T lymphocytes from chronic HBV infected patients.And PD-1 is negatively correlated with serum IFN-γ levels and positively correlated with serum IL-10 levels.The expression of PD-1 on the surface of CD8+ T lymphocytes influences the balance of helper T cell (Th)1/Th2,which might play a role in the persistence of HBV infection.