Objective To evaluate the clinical application of a muhiprobe FISH (multicolor fluorescence in situ hy-bridization,M-FISH) assay in voided urine specimens for detection of bladder cancer and compare the results with those afforded by urinary cytology.Methods Voided urine specimens from 100 cancer patients with or without surgery and 10 patients with prostate hyperplasia were obtained for urinary cytology and FISH.FISH was performed using a mixture of fluorescent labeler DNA probes for the centromeric regions of chromosomes 3,7,and 17 and 9p21 region.Cystoscopy with biopsy or tumor resection was performed in all patients and compared the pathological results with the cytological and FISH findings.Results The sensitivity of M-FISH were 75.6% for low grade tumors,100% for high grade tumors,with overall sensitivity of 85.5%.Urinary cytology affords an overall sensitivity of 62.9% (33.3% for low grade tumors,96.0% for high grade tumors).Significant difference in overall sensitivity was observed between M-FISH and urinary cytology (P＜0.05).The specificity of M-FISH and urinary cytology was 84.6% and 87.8% respectively.No significant difference in specificity was observed between M-FISH and urinary cytology.Conclusion M-FISH improves the sensitivity obtained with urinary cytology for bladder cancer detection with similar specificity,so can detect all high grade infiltrating tumors patients.

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

Natural polysaccharides are a kind of hydrophilic polymers.By grafting hydrophobic moieties to the polysaccharide backbone,could form self-assembled micelle with core-shell structure in aqueous by undergoing intramolecular or intermolecular association between hydrophobic moieties.The self-assembly micelles of amphiphilic polysaccharides provides a positive outlook for drug delivery systems with biodegradability,low antigenicity,good biocompatibility and so on.This review focus on biological activity of amphiphilic polysaccharide as self-assembly carriers,the principle of self-assembly of polymeric micelles,and the recent progress in hydrophobic modification of natural polysaccharides.

Objective To detect the expressions of Numb and α-catenin in the gastric cancer and matched normal gastric mucosa,and to analyze their relationship with clinicopathological factors of gastric cancer and explore their roles in gastric carcinogenesis and progression.Methods Immunohistochemical method was used to detect the expressions of Numb and α-catenin in 109 cases of tumor specimens and 97 cases of matched normal gastric mucosa.The correlations between Numb,α-catenin and clinicopathological factors were analyzed.Results The positive rates of Numb and α-catenin in gastric cancer were significantly lower than those in normal gastric mucosa (60.6％ ∶ 100.0％,x2 =48.361,P =0.000;76.1％ ∶ 100.0％,x2 =26.480,P=0.000).The expression of Numb protein was correlated with Lauren type (x2 =17.018,P =0.000) and tubular adenocarcinoma differentiation (x2 =17.586,P =0.000).The expression of α-catenin protein was correlated with Borrmann type (x2 =6.700,P =0.035),Lauren type (x2 =11.098,P =0.001),tubular adenocarcinoma differentiation (x2 =8.203,P =0.017) and lymph node metastasis (x2 =6.402,P =0.011).The expressions of Numb and α-catenin were statistically correlated in 109 cases of gastric cancer (rk =0.184,P =0.028).Conclusion Compared with normal gastric mucosa,both Numb and α-catenin expressions are down-regulated and the two expressions are correlated in gastric cancer.Numb and α-catenin may be involved in the regulation of histological differentiation of gastric cancer by certain passages,Numb protein may be adjusted by α-catenin pathway which is involved in Wnt-β-catenin pathway,and thus plays an important role in promoting cell proliferation,invasion and metastasis in human gastric cancer.

The flushing pump which is applied to clean operative wound has no temperature controlling function up to now, and doctors have to prepare the flushing fluid that has previously been warmed. The flushing pump system with medical constant temperature designed in our laboratory can absorb flushing fluid at the room temperature, and then eject flushing fluid with the temperature in accordance with the requirements of operations at a controlled constant flow rate. The system combines flow rate control with temperature control functions. The flushing pump system includes flushing part, temperature controlling part, key inputting part, liquid crystal displaying part and exceptional situation monitoring part. The present paper introduces the design method and principle of each part of the system at first, and then gives the debug method of all the system parameters. Finally the paper discusses the performance of the system according to the result of the experiment.
Equipment Design ; Equipment and Supplies ; Temperature

In surgery operations, wound should be cleaned with warm sterilized saline solution. In order to reach rapidly warming the washing solution from the room temperature during the surgery, we designed a thermostatic medical infusion pump. The present paper mainly presents researches on the two temperature control methods in the standby mode and in the flushing mode of the system. In the standby mode, the traditional proportional-integral-derivative (PID) control algorithm was adopted. In the flushing mode, dynamic characteristics of the system was changed in real time, which made the thermostatic control process more complex, and the fitted control function combined with the PID control algorithm was adopted in this mode. The temperature control parameters were adjusted in real time according to the initial temperature and the flow rate of the washing solution to obtain a constant temperature of the washing solution, no matter how the initial temperature and the flow rate are changed. The experiment results showed that this kind of control system performed well with a high accuracy.
Algorithms ; Equipment Design ; Infusion Pumps ; Sodium Chloride ; Solutions ; Temperature

BACKGROUND:Currently,there is not a standard method for in vitro culture and large scale amplification of umbilical cord blood mesenchymal stem cells(UCB-MSCs).OBJECTIVE:To investigate the isolation,purification and culture of UCB-MSCs in vitro,and to detect its surface marker variation.METHODS:The monocytes were harvested from UCB using 1.077 g/cm3 lymphocytes separating solution and density gradient centrifugation,followed by incubation in an incubator containing 5%CO2 at 37℃.The cell morphological changes were observed at different time points and the expression of surface marker was detected using flow cytometry.RESULTS AND CONCLUSION:The monocytes isolated from the UCB grew initially into numerous hematopoietic cell clones,most of which were granulocyte/macrophage colony-forming units and burst forming unit-erithroid,increasing by(37.1±2.3)and (10.4±1.7),respectively.Switzerland staining showed most of them were granulocyte clones(80,1±85.2)%,next was erythroid clones(14.2±1.8)%.At 7 days after culture,some shuttle fibroblast-like cells and fiat osteogenic-like cell spread the whole plastic well.At 14 days after culture,flow cytometry showed CD38+ cells accounted for 1.64%,and CD34+/CD38+ cells accounted for 1,71%,and CD34+/CD38- were 0.55%.PI+ and Annexin-V+ cells accounted for 0.05% and 0.18% respectively.At 21 days after culture,CD38+,CD34+/CD38+ and CD34+/CD38- cells were 74.32%,1.61%,and 0.24%.The results reveled that UCB-MSCs can be isolated and cultured in vitro.

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.

BACKGROUND: Stem cells are relatively primitive cells possessing the capabilities of self-renewal, high proliferation and multi-potential differentiation in vivo under certain conditions. Pancreatic stem cells and umbilical cord blood mesenchymal stem cells (MSCs) may serve therapeutic purpose clinically, but they are still difficult to culture in vitro at present.OBJECTIVE: To explore the method for isolation, purification and culture of pancreatic stem cells and umbilical cord blood MSCs in vitro and observe their morphological changes during culture in vitro.DESIGN: Completely randomized experiment with repeated measurement.SETTING: Stem Cell Research Center, Teaching and Research Division of Physiology, Medical School of Zhengzhou University.MATERIALS: This experiment was conducted in the Stem Cell Research Center, Teaching and Research Division of Physiology, Medical College of Zhengzhou University, between April 2004 and January 2005. Ten to fifteen newborn SD rats (1-3 days) were selected for culture in vitro of pancreatic stem cells, and fresh umbilical cord blood was collected from healthy woman (24-35 years old, with informed consent) at full-term delivery for culture in vitro of umbilical blood SMCs.METHODS: The abdomen of the newborn SD rat was opened under aseptic condition to obtain the pancreas, which was cut into small tissue blocks and digested with type-V collagenase for islet isolation. The isolated islets were purified in continuous roller-bottle culture. Umbilical cord blood was freshly collected for isolating the monocytes by means of density gradient centrifugation in lymphocyte separation medium (with density of 1.077 g/cm3). The islet cells and umbilical cord blood monocytes were cultured in the incubator at 37 ℃ with 5% CO2. The morphological changes of the cells were observed at designed time points and flow cytometry was used to determine the expression of cell surface molecules.MAIN OUTCOME MEASURES: The isolation and culture of pancreatic stem cells and umbilical cord blood MSCs, and their morphological changes during culture in vitro.RESULTS: During culture in vitro, the fusiform islet progenitor cells showed adherent polar growth and continuous proliferation, which covered the whole bottom of the flask after 12-14 days and could be subcultured for passages. However round cells appeared after removal of the growth factor and serum in the culture medium. The monocytes isolated from the umbilical cord blood grew initially into numerous hematopoietic cell clones, most of which proved to be granulocyte clones by Switzerland staining. Seven days later, flat flask wall-adhering epithelial cells and long fusiform fibroblasts were observed mixed with a number of osteoclasts. As the cell culture was prolonged, the cell number increased steadily.CONCLUSION: Pancreatic stem cells and umbilical cord blood SMCs can be cultured in vitro for further experiments.