Objective:To isolate the invasive and non-invasive cells from primary human renal cell carcinoma(RCC) in vitro.Methods: Fresh RCC surgical specimens from 32 primary RCC patients were primarily cultured following enzyme digestion or mechanical minimization in vitro.In vitro invasion assay using the Transwell cultures coating Matrigel was performed for separation and recovery of invasive and non-invasive cells from the primary culture of 3 RCC patients.The concentration of Matrigel,recovery time and trypsinization were subsequently optimized.Results: The successful rate of primary culture was(90.6%)(29/32).Recovery of invasive cells was performed ideally when matrigel(diluted into 1.0 mg/ml and 20 ?l) was coated onto the filter of the well;cell suspension was at a concentration of 5?10~(5)/ml and invasive cells were recovered on the 5th day of culture.The growth of non-invasive cells was scattered,while that of the invasive cells was focal.The doubling time of invasive cells was 36.1 h and that of non-invasive was 50.6 h.Conclusion: The in vitro invasion assay using the Transwell is able to separate and recover the highly invasive primary RCC cells.The primary cells represent intact subpopulation composition,but it can hardly get through the life span of human primary tumor cells.

Objective:To establish orthotopic mouse models of human renal cell carcinoma and to separate metastatic or non-metastatic renal cell carcinoma(RCC) from the same source.Methods: Surgical specimens/cell suspensions were transplanted into the various tissues of BALB/c nude mice(subcutis,cellular orthotopic injection into renal capsule,perinephrium,surgical orthotopic implantation into renal capsule).Tumorigenicity and metastasis were subsequently evaluated.Immunohistochemical analysis was employed to determine the expression of VEGF,bFGF,P16,Bcl-2 and C-met in the metastatic and non-metastatic RCC xenograft.Results: The incidences of tumorigenicity and metastasis of orthotopic model were the highest,being at 73.3%(11/15)and 20%(3/15),respectively.Compared with the non-metastastic RCC, VEGF expression was upregulated in the metastatic RCC(P

Objective To explore the factors related to the prognosis and survival duration of primary liver cancer patients after hepatectomy.Methods The data of primary liver cancer patientswho were treated by surgical resection were analyzed retrospectively.Kaplain-Meier method was used to evaluate survival rates.Log-rank test and Cox regression analysis were used to screen out related clinical phathology factors.Results The median survival time was eighteen months.Univarivate analysis showed that liver function Child-Pugh classification,cirrhosis,tumor size,HBV infection,AFP,portal vein tumor thrombus significantly correlated with survival rates (P ＜ 0.05).Multivariate analysis showed that liver function Child-Pugh classification,tumor size,AFP and portal vein tumor thrombus were the independent prognostic factors of primary liver cancer (P ＜ 0.05).Conclusion Many factors are related to the prognosis of primary liver cancer after operation.Liver function Child-Pugh classification,tumor size,AFP and portal vein tumor thrombus affect prognostic independently.

Vasculogenic mimicry (VM) can promote tumor growth, invasion, and metastasis. The formation of VM is regulated by various proteins including epithelial cell kinase, phosphoinositide-3 kinase, and hypoxia-inducible factor 1. Local microenvironment also plays an important role in regulating VM formation.

Hypoxia inducible factor (HIF-1α) is a hypoxia response regulator,which is highly expressed in malignant tumors.Researches have shown that HIF-1α plays an important role in epithelial mesenchymal transformation.It can regulate epithelial mesenchymal transition through a series of signal pathways and mechanisms at many levels,to participate in the invasion and metastasis of tumors.To explore the mechanism of HIF-lα controlling epithelial mesenchymal transformation has important significance for the development of new,effective tumor treatment methods.

ObjectiveStudy the relationship among CaSR expression, tubulointerstitial damage,metabolic disturbance of calcium and phosphorus and microvascular density around the tubulointerstitium in children with steroid-resistant nephrotic syndrome.Methods36 cases of children with primary nephrotic syndrome were divided into hormone-sensitive group and steroid-resistant group.Semi-quantitative scores for tubulointerstitial pathological evaluation of the extent of damage, automatic biochemical analyzer for the determination of serum calcium (Ca), phosphorus (P) concentration of renal tubular epithelial CaSR expression and microvessel microvascular density around the tubulointerstitium were determined by immunohistochemical assay.ResultsMore severe the tubulointerstitial damage, lower level of serum Ca and higher level of serum P were observed [(2.26 ± 0.15) mmol/L]in children of the steroid-resistant group and the steroid-sensitive group [(1.90 + 0.12) mmol/L, P ＜ 0.05].CaSR expression (4.63 + 0.78) of renal tubular epithelial cells in the steroid- sensitive group was significantly lower than that in the steroid-resistant group (6.56 + 1.22, P ＜ 0.05), but microvascular density was significantly higher in the steroid- sensitive group(2.98 +0.35 vs 2.02 +0.24, P ＜0.05).When the tubulointerstitial damage was mild, CaSR expression (4.15 +0.58) in renal tubular epithelial cells in the steroid- sensitive group (4.26 ±0.61) was lower than the steroid-resistant group(3.12 ± 0.33; 3.01 ± 0.21), and microvascular density was higher,but the difference was not significant(P ＞0.05).In the moderate tubulointerstitial damage, CaSR expression in renal tubular epithelial cells in the steroid- sensitive group (5.35 ± 0.64) was significantly lower than the resistant group (7.37 +0.81, P ＜0.01), and microvascular density was significantly higher than the resistant group (2.81 ±0.16, 2.02 ±0.14, P ＜0.05).Compared by mild and moderate tubulointerstitial damage in children with the steroid-resistant, CaSR expression (11.46 ± 1.38) in children with severe tubulointerstitial damage was significantly increased, and microvascular density (1.15 ± 0.11) was significantly decreased (all P ＜ 0.01).ConclusionsCaSR expression was increased and microvascular density around the tubulointerstitium was decreased in children with steroid-resistant nephrotic syndrome.Dut to steroid resistance, the cytotoxic of steroid damaged the renal tubular epithelial cells, the metabolic disturbance of calcium and phosphorus and the damage of blood vessel endothelium finally resulted in severe tubulointerstitial damage.

Organ-specific tumor cell adhesion to extracellular matrix (ECM) components and cell migration into host organs often involve integrin-mediated cellular processes. Direct integrin-mediated cell adhesion to ECM components in the space of Disse appears to be required for the successful liver metastatic formation of colon cancer. In the present study, human colon cancer HT-29 cells were transfected by liposome with integrin-beta(1) antisense oligodeoxynucleotide (ASODN). The integrin-beta(1) gene expression in HT-29 cells was significantly down-regulated. The migration of HT-29 cells was assayed using transwell cell culture chambers in vitro. The number of migrating HT-29 cells in experimental group was far less than that in control group (P<0.05). The models of hepatic metastasis in nude mice were established by the intrasplenic injection of transfected HT-29 cells. Thirty days later, the nude mice were killed and the average number of hepatic metastases (4.00+/-0.93 per mouse), average volume (10.10+/-6.50 mm(3) per mouse), average weight (0.0440+/-0.0008 g per mouse) in experimental group were remarkably reduced as compared with those in control group (P<0.05). Integrin-beta(1) expression in the hepatic metastasis was studied by immunohistochemistry (SP). Positive cell percentage of hepatic metastases in experimental group was markedly decreased as compared with that in control group (P<0.05). It was concluded that integrin-beta(1) may take part in hepatic metastasis, and down-regulation of integrin-beta(1) expression may play a key role in decreasing migration and hepatic metastasis of human colon carcinoma cells (HT-29).

Objective To explore the applied value of thyroid imaging reporting and data system (TI-RADS) strati?fication in diagnosing thyroid nodules in patients with different ages.Methods A total of 527 patients with thyroid nod?ules (aged 26-65) were divided into four groups:aged 26-35 group, aged 36-45 group, aged 46-55 group and aged 56-65 group. TI-RADS was used to evaluate patients in four groups. The results of TI-RADS were compared with the pathologic di?agnostic analysis. Results For the low aged groups (aged 26-35 group and aged 36-45 group), the positive predictive val?ue, the negative predictive value and accuracy rate were more than 90%. With regard to the patients aged 46-55, the positive and negative predictive values and accuracy rate were 81.5%, 87.5%, and 81.9%, respectively. The positive and negative predictive values, accuracy rate were 76.6%,88.8%and 77.9%for patients aged 56-65 group. The diagnostic rate of TI-RADS in patients with thyroid lesions was lower in aged 56-65 group than that of other groups. Conclusion TI-RADS method is affected by age. The diagnostic rate is lower in the high age group than that of the low age group.

The research and quality control of traditional Chinese medicine has meaningful importance, because it has influence not only in the health and treatment of patients, but also in the solid growth and development of phar-maceuticals companies. In some cases, for the complex of TCM, the common QC method on single or multi-target compounds can't really and truly disclose the quality of the Chinese materia medica. Therefore, a lot of researchers do plenty of works to make clear the effectiveness basis, to improve the quality and realize the modernization of TCM. All of these works close together with modern analysis and separation technology. In this article, a novel analy-sis technology-UltraPerformance Convergence Chromatography (UPC2) based its characters and applications should be introduced. It should be a helpful technology for the TCM researchers to facilitate the study and QC works.

Objective To measure the concentration of serum transthyretin (TTR) of patients with different stages of diabetic retinopathy (DR).Methods A total of 176 patients with diabetes mellitus were included in this study.There were 104 males and 72 females.The patients aged from 21 to 74 years,with the mean age of(56± 11) years.The diabetes duration raged from 1 to 30 years,with the mean diabetes duration of (10 ± 7) years.The HbA 1C was 5.2％-14.1％,with the mean HbA 1C of (8.6 ± 2.0)％.According to the fundus examination,58 patients had DR (33.0％),but the other 118 patients not (67.0％).For these DR patients,10 patients were in stage Ⅰ (5.7％),26 patients in stage Ⅱ (14.8％),8 patients in stage Ⅲ (4.5％),and 14 patients in stage Ⅳ (8.0％).The concentration of serum TTR was measured by enzyme-linked immunosorbentassay kit.The differences in the concentration of serum TTR between different DR stages were compared.Bivariate analysis was used to analyze the influencing factors of TTR.Results The concentrations of serum TTR of the patients without DR or with DR of stage Ⅰ to Ⅳ were (224.96±65.47),(383.68± 102.99),(247.44±63.21),(228.2 ± 45.89),(189.34± 70.12) mg/L,respectively.The difference between different DR stages was statistically significant (F=14.690,P＜ 0.001).Bivariate analysis showed that the concentration of TTR was correlation to DR (r=0.179,P=0.017).There was no correlation between the concentration of TTR and diabetes duration (r=-0.027,P=0.727),hypertension (r=0.018,P=0.810),hyperlipoidemia (r=0.101,P=0.182),and the use of insulin (r=-0.032,P=0.675).Conclusion The concentration of serum TTR was increased in early DR patients,and gradually decreased with the progression of DR.The concentration of TTR is correlated to DR.