Objective To investigate the effect of over-expressed Cyr61 on the expression of extracellular matrix of human renal tubular epithelial cells(HKC), and explore the role of Cyr61 in the pathogenesis and development of autosomal dominant polycystic kidney disease(ADPKD). Methods Cyr61 gene cDNA was amplified by RT-PCR. A recombinant plasmid pcDNA3.1+ Cyr61 was constructed by cloning Cyr61 gene into pcDNA3.1. HKC cells were transfected with wild-type Cyr61 plasmid vector (pcDNA3.1+Cyr61). Then the fusion of Cyr61 gene and expression of its protein were detected by RT-PCR and Western blot. The mRNA expression of Cyr61, collagen Ⅰ, collagen Ⅳ, and laminin were determined in four groups (the untransfected cells, pcDNA3.1 transfected cells, pcDNA3.1+Cyr61 -transfected cells and cyst-lining epithelial cells) by fluorescence quantum PCR. Results The amount of Cyr61 protein in Cyr61-transfected cells and cyst-lining epithelial cells were much greater than the control. The mRNA expression of Cyr61, collagen Ⅰ, collagen Ⅳ and laminin in Cyr61-transfected cells were all amplified significantly, and the level of collagen Ⅳwas much higher than collagen I(P

Objective To investigate the effect of cysteine-rich protein 61 (Cyr61) on proliferation and cell cycle in human renal tubular epithelial cells (HK-2).Methods Cyr61 cDNA was cloned into pEGFP-N2,then HK-2 cells were transfected with the recombinant plasmid pEGFP-N2-Cyr61 by Lipofectamine.The cell proliferation was measured by MTT.The expression level of Cyr61,p-FAK and cyclin dependent cyclin-dependent kinase 2 (CDK2) protein were detected by Western blotting.The cell cycle and cell apoptosis were analyzed by flow cytometry.Results The recombinant plasmid pEGFP-N,-Cyr61 could be transfected into HK-2 efficiently.After transfection,the proliferative activity was significantly increased,the proportion of HK-2 cells in G1 phase decreased and in S-phase increased significantly,the level of cell apoptosis decreased markedly (all P ＜ 0.01).The expressions of Cyr61,p-FAK and CDK2 in Cyr61-transfected group were all amplified significantly (all P ＜ 0.01).Conclusions Cyr61 protein over-expressed in HK-2 cells can increase CDK2 expression throngh FAK pathway,resulting in the promotion of HK-2 cells entering into S phase,cell proliferation and the reduction of cell apoptosis.

Objective To investigate the effect and mechanism of cysteine-rich protein 61 (Cyr61) on oxidative stress in human kidney tubular epithelial cell line after anoxia.Methods Human kidney tubular epithelial cell line (HK-2 cells) were divided into 5 groups:control group,Cyr61 group,MAPK inhibitor group (Cyr61 +PD98059),p38 inhibitor group (Cyr61 +SB203580) and PI3K inhibitor group (Cyr61+Wortmannin).Each group was pretreated for 12 h and then injured by anoxia.The cell viability was determined by MTT assay and the apoptosis rate of HK-2 cells was determined by flow-cytometry.The cellular ROS level was measured by spectro-fluorometry.The cellular superoxide dismutase (SOD) and catalase (CAT) were measured by nephelometry test.The expression of Nrf2 in HK-2 cells was detected by Western blotting.Results Anoxia enhanced the expression of ROS and Nrf2,decreased the expression of SOD and CAT significantly,meanwhile decreased HK-2 viability and increased HK-2 apoptosis (all P ＜ 0.05).Cyr61 increased the expression of p-Akt,Nrf2,SOD and CAT in HK-2,and decreased the expression of ROS,at the same time increased HK-2 viability and decreased HK-2 apoptosis (all P ＜ 0.05).Wortmannin inhibited the expression of p-Akt,Nrf2,SOD and CAT,meanwhile decreased HK-2 viability and increased HK-2 apoptosis (P ＜ 0.05).PD98059 and SB203580 had no affect on HK-2 compared to Cyr61 group (P＞0.05).Conclusions Cyr61 promotes the expression of Nrf2 through PI3K pathway in HK-2,which enhances the expression of SOD and CAT,and decreases the expression of ROS.Cyr61 exhibits protective effects on HK-2 cells injured by oxidative stress after anoxia.

Objective To investigate the relationship between various clinicopatholngic factors and prognosis of early gastric cancer. Methods The clinical data of 459 patients with early gastric cancer who had undergone gastrectomy combined with extended (D2) lymphadenectomy at Zhongshan Hospital from January 2002 to October 2007 were retrospectively analyzed. Survival was calculated using the Kaplan-Meier method, and clinicopathologic factors such as age, sex, tumor size, gross morphology, tumor differentiation, depth of invasion, lymphatic vessels involvement and lymph node metastasis were analyzed using the Cox regression model. Univariate analysis was done by Log-rank test. Results Univariate analysis demonstrated that tumor size and differentiation, depth of invasion, lymphatic vessels involvement and lymph node metastasis significantly affected survival in patients with early gastric cancer (χ~2 = 8.476, 6.210, 4.014, 14. 197, 55.027, P < 0.05). The status of lymph node metastasis was an independent predictor of survival in patients with early gastric cancer, and the more metastatic lymph nodes detected, the greater the influence on the prognosis. Conclusions Lymph node metastasis is the most important factor influencing the prognosis of early gastric cancer. Appropriate lymph node dissection is necessary, especially for patients with risk factors associated with lymph node metastasis.

Objective To prepare and identify monoclonal antibody against LRR-WSC domain of polycystin-1 and to investigate the distribution of polycystin-1 in kidney tissues and kidney cell lines. Methods BALB/c mice were immunized with fusion protein PC1-e of polycystin-1 LRR-WSC domain. The splenocytes were fused with myeloma cells by PEG 4000 and the hybridomas were selected in HAT medium. The hybridoma clones secreting antibodies against polycystin-1 LRR-WSC domain were detected by enzyme-linked immunosorbent assay ( ELISA) and cloned by limiting dilution. The specificity of anti-polycystin-1 LRR-WSC domain monoclonal antibody from hybridoma was verified by ELISA and Western blot. The distribution of polycystin-1 in tissues and cells was detected by immunohistochemical method. Results One cell line of hybridoma secreting monoclonal antibody against polycystin-1 was established. Western blot analysis showed that the monoclonal antibody reacted strongly and specifically to polycystin-1 LRR-WSC domain. Distribution of polycystin-1 in fetal kidney was localized in tubular epithelium. In normal adult kidney tissues, our study showed that polycystin-1 was mainly expressed in the medullary collecting ducts and distal convoluted tubules. Positive staining was also found in the majority of cyst-lining epithelial ceEs of cystic tissue from autosomal dominant polycystic kidney disease ( ADPKD) patients. Expressions of polycystin-1 were found in either ADPKD cyst-lining epithelia cell line and LLC-PK1, clearly plasma membrane and intracytoplasmic staining of polycystin-1 were observed. Conclusion Specific monoclonal antibody against polycystin-1 LRR-WSC domain were obtained. The antibody is important to researching the mechanism of ADPKD. The distribution of polycystin-1 in kidney tissues and cells show that polycystin-1 was important in tubular elongation and the maintenance of tubular architecture.

Objective To evaluate the diagnostic value of spiral CT [CT arterial portography (CTAP) and CT hepatic arteriography(CTHA)] for hepatic carcinoma. Methods The CTAP and CTHA manifestations in 21 patients with hepatocelluar carcinoma were analyzed and compared with those in three-phase enhanced CT . Results The detection rate of three-phase enhanced CT was 72.4% , CTAP and CTHA was 96.1% and 88.2% respectively. The detection rate of combination CTAP and CTHA was 98.7% . They could reveal tumors of 0.5 cm in diameter . Both CTAP and CTHA gave false-positive findings , such as perfusion defects in 22.3% of CTAP and non-pathologic enhancement in 30.2% of CTHA .Conclusion With the use of spiral CT technique , the quantity of contrast material administrated in CTAP and CTHA can be considerably reduced . And the quality of CT images significantly improved . The detection rate of CTAP and CTHA is higher than that of three-phase enhanced CT . Simultaneous use of both procedures may decrease the false-positive rate.

Objective:To combine the detection of serum levels of adenosine deaminase (ADA) and T-cell spot test (T-spot.TB),and to explore their significances in diagnosis of pulmonary tuberculosis.Methods:159 patients suspected with pulmonary tuberculosis were selected and divided into pulmonary tuberculosis group and non-tuberculosis group (n=68);80 healthy people were randonly selected as healthy control group.The serum ADA levels and number of T-spot of the subjects in three groups were detected.Ther serum ADA levels and the positive rates of T-spot.TB in various groups and their sensitivities and specifities were compared. Results:The serum ADA level of the patients in pulmonary tuberculosis gruop was (22.10±6.60)U·L-1;those in non-tuberculosis group and healthy control group were (16.90±6.35)and (8.70±5.98)U·L-1;the serum ADA level in pulmonary tuberculosis group was significantly higher than those in non-tuberculosis group and heathy control group (P<0.05).The positive rate of serum ADA level in diagnosis of pulmonary tuberculosis was 56% and the T-spot.TB positive rate in diagnosis of pulmonary tuberculosis was 87.9%. Combined use of parallel test, the detection sensitivity was 91.2%;using the series of joint tests,the specificity was 94.6%.Conclusion:Combined detection of serum level of ADA and T-spot.TB can significantly improve the clinical diagnosis efficacy of pulmonary tuberculosis.

Objective:To investigate characteristic expression of TGF-?1 in human ADPKD and clarify its role in the development and progression of human ADPKD. Methods:Cyst fluid, urinary and plasma TGF-?1 levels were determined by ELISA in 39 ADPKD patients. The results were compared with those of normal subjects and of patients with simple renal cyst. TGF-?1, TGF-?1 receptors types Ⅰ, types Ⅱ, CTGF mRNA and proteins in the kidneys of human ADPKD were examined by in situ hybridization and immunohistochemistry. The relationship of the above fibrosing-associated indicators with the degree of interstitial fibrosis was analyzed. Results:Plasma TGF-?1 level was the highest among body fluids. In ADPKD and simple renal cyst, TGF-?1 levels were significantly higher than those in normal subjects (15.12 ?8.53)?g/L vs (5.41?1.31) ?g/L,P

Objective To investigate the risk factors for contralateral hematoma progression of bilateral chronic subdural hematomas (bCSDHs) after initial unilateral evacuation,and finally develop a prognostic scoring system.Methods Sixty-one patients with bCSDHs underwent initial unilateral evacuation in our hospital from October 2012 to March 2019 were chosen in our study.During follow up,CT examination was used to determine whether the patients had developed eontralateral hematoma.The clinical data of patients from the progressive group and non-progressive group were retrospectively analyzed and compared.Multivariate Logistic regression was used to analyze the independent risk factors for postoperative contralateral hematoma progression.Receiver operating characteristic (ROC) curve was established to predict the progression of contralateral hematoma for each risk factor.A prognostic grading system was developed on the basis of independent risk factors and cut-off value.All patients were scored according to the scoring system and the progression rate of different scores were re-analyzed.Results As compared with the non-progressive group,the progressive group had higher proportions of patients with extensive contralateral hematoma distribution or low density of contralateral hematoma,higher amount of preoperative contralateral hematoma and postoperative hematoma,with statistically significant differences (P＜0.05).Multivariate Logistic regression analysis showed that extensive contralateralhematomadistribution (OR=16.726,95％CI:2.034-137.557,P=0.009),and hematoma volume after contralateral hematoma surgery (OR=1.044,95％CI:1.012-1.078,P=0.007) were independent risk factors for contralateral hematoma progression.ROC curve showed that areas under the curve for contralateral hematoma distribution and hematoma volume after contralateral hematoma surgery were 0.682 and 0.737,respectively.Limited type of contralateral hematoma was set as 0 score,and extensive type was set as one score;hematoma volume after contralateral hematoma surgery＞40 cm3 was set as one score,and that≤40 cm3 was set as 0 score.All patients were scored,and the scoring system was ranged from 0 to 2 scores;the contralateral hematoma progression rate of 0,1,and 2 were 0％,23.81％,and57.69％,respectively,with significantdifferences (P＜0.05).Conclusion Extensive contralateral hematoma distribution and hematoma volume after contralateral hematoma surgery are independent risk factors for contralateral hematoma progression of bCSDHs after initial unilateral evacuation;the prognostic scoring system is simple and practical,which can serve as part of clinical references.

OBJECTIVETo explore the risk factors associated with the prognosis in poorly differentiated(G3) advanced gastric cancer (AGC) and the effect of number of harvested lymph nodes on the prognosis.

METHODSClinical data of 484 patients with G3 advanced gastric cancer undergoing gastrectomy combined with lymphadenectomy in Zhongshan Hospital from December 2002 to October 2007 were retrospectively analyzed. The χ(2) test or Fisher's exact probability method was used to test measurement data. Survival was calculated using Kaplan-Meier method. Clinicopathological factors such as age, invasion depth, number of harvested lymph nodes, lymph node metastasis, ratio of metastatic lymph node, lymphatic vessels involvement were analyzed using the Cox regression model.

RESULTSNumber of lymph node dissected, lymph node metastasis, ratio of metastatic lymph node and lymphatic vessels involvement were significantly affected by groups of 15 lymph node dissected in G3 AGC patients (P<0.05). Invasion depth, number of lymph node dissected, lymph node metastasis, ratio of metastatic lymph node and lymphatic vessels involvement were significantly independent predictors of survival in G3 AGC patients (all P<0.05). Patients with at least 20 lymph nodes had a better survival rate than those with less than 20(P<0.01).

CONCLUSIONSInvasion depth, number of harvested lymph nodes, lymph node metastasis, ratio of metastatic lymph node and lymphatic vessels involvement are important factors influencing the prognosis of G3 AGC. At least 20 lymph nodes should be recommended to ensure the quality of lymphadenectomy for gastric cancer.

Gastrectomy ; Humans ; Lymph Node Excision ; Lymph Nodes ; Lymphatic Metastasis ; Prognosis ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; pathology ; surgery ; Survival Rate