ObjectiveTo study the correlation between preoperative serum angiopoietin-2 (Ang2) levels and lymph node metastasis in early gastric cancer(EGC).MethodsPreoperative serum Ang-2 and CEA levels in 62 patients with EGG and 30 normal controls were measured by ELISA technique respectively.The metastasis of lymph node in these cases were determined by HE staining.The relation between preoperative serum Ang-2 levels and lymph node metastasis in EGC and pathologic characterization was investigated.ResultsCompared with normal controls,preoperative serum Ang-2 levels in EGC group were significantly elevated [( 282.5 ± 110.6 ) μg/L vs ( 187.4 ± 32.7 ) μg/L,P ＜ 0.01].Preoperative serum Ang-2 levels in node-positive gastric cancer group was significantly higher than that in the node-negative gastric cancer group [(34 7.2 ± 79.5 ) μg/L vs (265.6 ± 90.2) μg/L,P ＜ 0.01],while CEA levels changed a little.Preoperative serum Ang-2 levels in the EGC group presented marked correlation with the differentiated degree of the tumor tissue,tumor size,depth of tumor invasion but it had no correlation with the histological type.Preoperative serum Ang-2 levels were an independent high-risk factor of lymph node metastasis in EGC according to multivariate logistic regression analysis.ConclusionsPreoperative serum Ang-2 levels may be a good clinical predictor for lymph node metastasis of EGC,which is useful for the treatment direction.

Objective To investigate the correlation between expression of human telomerase reverse transcriptase (hTERT) mRNA in peripheral blood and postoperative recurrence and prognosis in the patients with advanced colorectal cancer (CRC).Methods The preoperative and 1 month postoperative quantitative expression of hTERT mRNA in peripheral blood in 85 patients with advanced CRC were detected with real-time fluorescence quantitative real-time fluorescence quantitative polymerase chain reaction (RTPCR).The quantitative expression of hTERT mRNA in peripheral blood in 30 normal controls were also detected.The postoperative prognosis was followed-up,and the quantitative expression of hTERT mRNA in peripheral blood in the patients with recurred CRC and the patients without recurred CRC were detected repeatedly.The factors that affected the preoperative hTERT mRNA expression were analyzed.The relationship was investigated between the hTERT mRNA expression levels and postoperative recurrence and prognosis in the patients with advanced CRC.Results The hTERT mRNA expression levels in peripheral blood in advanced CRC group were significantly higher than those in the normal controls [(6.22 ±5.85)2-△△Ct vs (0.45 ±0.27)2-△△Ct,(P ＜0.01)],and were significantly decreased at the time point of 1 month after the operation (P ＜ 0.01).The hTERT mRNA expression levels in the postoperative recurred CRC group were significantly higher than those in the 1 month postoperation group and no recurrence CRC group [(5.07 ± 2.87)2-△△Ct vs (1.83 ± 1.08)2-△△Ct,(2.15 ± 1.49)2-△△Ct,(P ＜0.01)].Univariate analysis revealed that preoperative high hTERT mRNA expression levels in peripheral blood were related to the differentiated degree of tumor,tumor size,lymph node metastasis ratio,and liver metastasis (P ＜ 0.05 or P ＜ 0.01).Multivariate logistic regression analysis revealed that differentiated degree of the tumor,lymph node metastasis,and liver metastasis were independent risk factors for the elevation of preoperative hTERT mRNA expression in the advanced CRC.Elevated preoperative hTERT mRNA expression was positively related to the postoperative recurrence rate,and was negatively related to postoperative survival rate.Conclusions Preoperative hTERT mRNA expression levels in peripheral blood were related to the important pathologic features in patients with advanced CRC,and might be a valuable prognostic factor.Postoperatively elevated hTERT mRNA expression in peripheral blood was related to recurrence of advanced CRC.

To investigate the role of progesterone receptor (PR) expression in malignant melanoma (MM), PR and proliferative cell nuclear antigen (PCNA) expression were immunohistochemistrically evaluated in a series of 35 specimens of MM, and the correlation between the immunohistochemistrical findings and clinicopathological data was also analyzed. PR expression was detected in 25.7% (9/35) of the patients with MM. No PR expression was observed in nevi. PR expression was inversely correlated with PCNA expression (r=-0.353, P=0.026). PR expression was slightly increased in females, subjects aged under 55 y, those with ulceration, non-acral subtype and diagnosis delay longer than 1 y, but the difference was not statistically significant. Selective expression of progesterone receptor in malignant melanoma might be correlated with inhibited tumor growth.
Gene Expression Regulation, Neoplastic ; Immunohistochemistry ; Melanoma/*metabolism ; Models, Biological ; Prognosis ; Proliferating Cell Nuclear Antigen/*metabolism ; Receptors, Progesterone/*biosynthesis ; Receptors, Progesterone/genetics ; Skin/metabolism ; Skin Neoplasms/metabolism

This study investigated the growth-regulating effects of progesterone (Prog) on nPR-negative malignant melanoma cells and the possible mechanisms. A375 and A875 cells were cultured and treated with Prog of different concentrations. For signal transduction pathway studies, the cells were pretreated with Prog receptor antagonist (RU486, 1x10(-7) mol/L) or MAPK inhibitor (U0126, 5x10(-6) mol/L) for 1 h and then co-incubated with prog (10(-9) mol/L) for another 24 h. Indirect immunofluorescence assay, MTT, flow cytometry and Western blotting were used for assessing the nPR expression, cell growth, cell apoptosis and ERK1/2 Phosphorylation, respectively. Our results showed that lower progesterone concentration promoted the proliferation of both A375 and A875 cells, but this growth-stimulatory effect decreased at progesterone concentration of 1x10(-7) mol/L or higher. The response could be abolished by MAPK inhibitor U0126, but could not be blocked by progesterone antagonist RU486. Flow cytometry exhibited that high concentration ([Symbol: see text]1 x 10(-7) mol/L) progesterone increased the apoptosis of the two cells in a dose-dependent manner. The level of ERK1/2 phosphorylation was increased by a lower progesterone concentration, but reduced by a higher concentration (1x10(-6) mol/L). These results suggest progesterone exerts growth-regulating effects on nPR-negative tumor cells through a non-genomic mechanism.

Objective To study the relationship of symptoms of female gonococcal infections to Chlamydia trachomatis infection, serum sex hormone levels, etc. Methods A total of 136 gonorrhea female patients without obvious symptoms were recruited in this study together with 45 gonorrhea patients with obvious symptoms as the controls. Serum progesterone (P) and estradiol (E2) levels were measured by radio immunoassay (RIA). Cervical swabs were obtained from the subjects and eluted into isotonic saline solution, the elution was divided into 2 portions and tested for the levels of TNF-α and IL-1β by ELISA and for the DNA of C. Trachomatis and N. Gonorrhea with PCR. Statistical analysis was carried out by SPSS for Windows (version 12.0). Results There was no statistical correlation between C. Trachomatis infection and asymptomatic status of female gonococcal infection (χ2 = 0.016, P > 0.05). However, the decrease in the level of TNF-α and IL-1β significantly correlated with the increase in serum progestogen (r = -0.8798, -0.8935, respectively, both P < 0.01). Conclusion The high serum level of progesterone may be associated with the asymptomatic status of gonococcal infection.

Objective To study the difference and clinical significance of serum Angiopoietin-2 (Ang-2) level in gastric cancer, precancerous disease and the changes in post operation follow-up patients.MethodsSerum Ang-2 and CEA levels were detected with ELISA method in 158 gastric cancer patients, 31 chronic astrophic gastritis patients, 38 gastric adenomatous polyp patients, 28 chronic gastritis patients with benign gastric ulcer and 30 healthy controls.The changes of serum Ang2 levels in 131 patients with radical gastrectomy were measured in 2-year post-operational followed-up and the prognosis was also evaluated.The correlation between serum Ang-2 level and the pathologic character of gastric cancer was analyzed.ResultsThe serum Ang-2 and CEA levels of gastric cancer group were (331.8± 64.3) μg/L and (42.6 ± 37.3) μg/L, respectively, which were significantly higher than those of healthy control group [(187.4±32.7) μg/L and (4.2±3.1) μg/L]and benign gastric ulcer group [(197.3±35.4) μg/L and (4.5±3.2) μg/L](all P ＜0.01).The serum Ang-2 level in chronic astrophic gastritis group was also markedly increased (P＜0.05), however there was no significant change in CEA level.There was no obvious change of Ang-2 and CEA level in gastric adenomatous polyp group (P＞0.05).Pre-operation serum Ang-2 levels of the relapse group and the metastasis group were significantly higher than that of no relapse group (P＜0.05).Post-operation serum Ang-2 level of no relapse group decreased to normal range in 1 month and without obvious increase in 2 years follow-up.The serum Ang-2 level of the relapse group also decreased to normal range in 1 month post-operation, while which was significantly higher than that of no relapse group and normal control group from 6 months to 2 years after operation (P＜0.01).Serum Ang-2 level of the metastasis group was markedly higher than that of no relapse group and normal control group during 2 years after operation (P＜0.01).The 2-year survival rate of no relapse group was significant higher than that of relapse group and metastasis group.The change of serum Ang-2 level was correlated with in the gastric cancer differentiation degree, TNM pathologic classification, lymphatic metastasis, distant metastasis, the depth of invasion and the tumor size (P＜0.01), however not correlated with histopathology type and the location of tumor.ConclusionSerum Ang-2 level may be a new and valuable gastric cancer marker, which will help to monitor tumor relapse, migration and prognosis assessment.

Objective To study the differential diagnostic value of soluble epithelial cadherin (sE-cad) in malignant and benign ascites. Methods The concentrations of sE-cad and carcino-embryonic antigen (CEA) in abdominal cavity synovia patients with benign diseases (31 cases), benign ascites(82 cases) and malignant ascites(119 cases) were ineasured by ELISA technique. Results The concentrations of sE-cad and CEA in malignant ascites patients were significantly higher than those in benign ascites patients and abdominal cavity synovia patients (P < 0.01). The sensitivity, negative predictive value and totally accurate rate of sE-cad was 89.9%, 86.8% and 92.5% respectively,which was significantly higher than those of CEA(68.1%, 65.8% and 76.6%)(P < 0.05). sE-cad in malignant ascites showed a certain tissue specificity. Conclusion sE-cad is an useful tumor marker in differential diagnosis of benign and malignant ascites.

Objective To investigate the role of myeloid dendritic cells(mDC)in the pathogenesis of ankylosing spondylitis (AS),and to study the mechanism of tumour necrosis factor (TNF)-α blocker on the treatment of AS by counting the number of mDC before and after the treatment.Methods Peripheral blood from 21 AS patients treated with 50 mg rhTNFR-Fc and 15 AS patients treated with placebo were investigated at week 0,week 2 and week 6 in a randomized,double-blind,placebo-controlled trial.Three-color flow cytometry analysis was used to investigate the change of the number of mDC before and after the treatment.And their correlation with the clinical parameters was analyzed.Results MHC Class Ⅰ positive mDC(Lin-/CD11c+/HLA-abc+)in AS patients was slightly less than in healthy controls (not statistically significant).No significant change in MHC Class Ⅰ mDC number was observed after drug treatment.The number of MHC Class Ⅱ mDC cells was not correlated with clinical parameters.Conclusion The treatment with rhTNFR-Fc in AS induces a significant upregnlation of MHC-Ⅱ DC.

Objective Clinical treatment can delay the development of renal interstitial fibrosis , but it can not reverse renal dysfuntion.The article was to discuss the influence of recombinant human erythropoietin ( rHuEPO ) on inflammatory factors in the process of renal interstitial fibrosis and its possible mechanism . Methods The vitro cultured HK-2 cells were randomized into 7 groups:the blank control group , rHuEPO control group ( addition of 20U/mL rHuEPO), albumin stimulation group (addition of 5mg/mL albumin), 5mg/mL rHuEPO intervention group (5mg/mL albumin +5U/mL rHuEPO), 10 U/mL rHuEPO intervention group (5mg/mL albumin +10 U/mL rHuEPO), 20U/mL rHuEPO intervention group (5mg/mL albumin +20U/mL rHuEPO), and Rho inhibi-taion group (addition of 5mg/mL albumin 30min after 10μmol/L Y27632), 24 h acting time for each group.We observed the changes of cell morphology in each group .Reverse transcription polymerase chain reaction ( RT-PCR) was used to evaluate the mRNA levels of RhoA, ROCK1 and IL-6 , and ELISA was applied to measure the levels of supernatant TNF-αand IL-6 protein. Results The form of pebbles or paving stone was observed in blank control group and rHuEPO intervention groups , a long and thin spindle change with the appearance of fibre cells in albumin stimulation group , the transformation to pebbles in 5, 10, 20 mg/mL rHuEPO intervention groups , the form of oval and slightly increased intercellular space in Rho inhibitaion group .Compared with the blank control group , the expressions of RhoA mRNA, ROCK1 mRNA and IL-6 mRNA significantly increased in the albumin stimulation group (P<0.05), while significantly reduced in 5, 10, 20 mg/mL rHuEPO intervention groups (P<0.05), which was in negative relation with the rHuEPO concentrations .Compared with the albumin stimulation group , the expressions of ROCK 1 mRNA and IL-6 mRNA reduced in Rho inhibtation group (P<0.05), while there was no significant difference as to the expression of RhoA mRNA .ELISA results showed:compared with blank control group , the expressions of supernatant TNF-α([452.32 ±33.23] ng/L vs [1347.54 ±41.52] ng/L), IL-6 protein([884.62 ±0.73] pg/L vs [95.12 ±0.32]pg/LP<0.05) increased significantly.Compared with albumin stim-ulation group, the expressions of TNF-αin 5, 10, 20 mg/mL rHuEPO intervention groups and Rho inhibitation group reduced signifi-cantly([1003.32 ±3.42] ng/L, [821.32 ±21.32] ng/L, [590.15 ±7.68] ng/L, [488.13 ±65.03] ng/L vs [1 347.54 ± 41.52]ng/L,P<0.05), while the expressions of IL-6 mRNA reduced accordingly in 5, 10, 20 mg/mL rHuEPO intervention groups and Rho inhibitation group reduced significantly ([656.68 ±0.55] pg/L, [422.35 ±0.22] pg/L, [217.32 ±0.35] pg/L, [309.49 ±0.21] pg/L vs [884.62 ±0.73]pg/L,P<0.05).Moreover, there was significant statistical difference among 5, 10, 20 mg/mL rHuEPO intervention groups(P<0.05). Conclusion RHuEPO can inhibit the transdifferentiation process of HK-2 cells in-duced by albumin by suppressing inflammation factors , and the mechanism may be involved in RhoA/ROCK signaling pathway .

Objective To investigate the correlation of the expression of human telomerase reverse transcriptase (hTERT) mRNA in peripheral and portal vein blood to liver metastasis and prognosis of patients with colorectal cancer (CRC).Methods From January 2009 to April 2011,a total of 181 patients diagnosed as primary CRC and received radical resection were enrolled.The relative quantitative expression of hTERT mRNA in preoperative peripheral blood and intraoperative portal vein blood were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR).All the patients were followed up for three years after operation.The relative quantitative expression of hTERT mRNA in peripheral blood before operation and intraoperative portal vein blood were compared between patients with synchronous liver metastasis (18 cases) and without synchronous liver metastasis (163 cases); between metachronous liver metastases (29 cases) and without metachronous liver metastases (152 cases).The relationship between the expression of hTERT mRNA in intraoperative portal vein blood and clinicopathological features of patients with CRC was analyzed.The t-test was used.Univariate and Multivariate Cox analysis were used for risk analysis of metachronous liver metastases.Log-rank test was used for comparisons of survival rate between the group with high hTERT mRNA expression in intraoperative portal vein blood during operation and the group with low expression.Results The relative quantitative expression of hTERT mRNA in preoperative peripheral blood and intraoperative portal vein blood in synchronous liver metastases group (8.04±3.79 and 11.88±4.19) was higher than that of no synchronous liver metastases group (4.30±2.81 and 4.94±3.37,t=5.159 and 8.084; both P＜0.01).The relative quantitative expression of hTERT mRNA in preoperative peripheral blood and intraoperative portal vein blood in metachronous liver metastases group (7.16±3.08 and 9.83 ± 2.96) was higher than that of no synchronous liver metastases group (4.11±2.58 and 4.56±3.09,t=5.648 and 8.467;both P＜0.01).The hTERT mRNA expression in intraoperative portal vein blood of patients with CRC changed significantly with the differentiated degree of tumor,tumor size,different invasion degree of tumor,lymph node metastasis,recurrence after operation,survival time (t =2.987,2.281,2.135,5.070,5.431 and 6.803,all P＜0.05).Univariate analysis revealed that the expression of hTERT mRNA in preoperative peripheral blood and intraoperative portal vein blood both were correlated with postoperative metachronous liver metastases (x2=9.522 and 16.393,both P＜0.01).Multivariate Cox analysis showed that both of them were independent risk factors (relative risk (RR)=4.286 and 9.783).The two-and three-year survival rates of the patients with high hTERT mRNA expression levels in intraoperative portal vein blood were 64.6 ％ and 52.3 ％,and those of low expression group were 91.4 ％ and 85.3 ％,and the differences between two groups in two-and three-year survival rate were statistically significant (x2 =5.313,P＜0.05; x2 =8.925,P＜0.01).Conclusions The expression of hTERT mRNA in intraoperative portal vein blood was closely correlated with the important pathologic features,prognosis and liver metastasis of patients with CRC and its predictive value of postoperative CRC metachronous liver metastases was higher than the expression of hTERT mRNA in preoperative peripheral blood.The hTERT mRNA in intraoperative portal vein blood can be a postoperative prognostic marker of patients with CRC.