Response of CRH system in brain and gill of marine medaka to seawater acidification.

Corticotropin-releasing hormone (CRH) is mainly secreted by the hypothalamus to regulate stress when environmental factors change. Gills contact with water directly and may also secrete CRH to maintain local homeostasis. Ocean acidification changes water chemical parameters and is becoming an important environmental stressor for marine fish. The response of brain and gill CRH systems to ocean acidification remains unclear. In this study, marine medaka were exposed to CO2-acidified seawater (440 ppm, 1000 ppm, and 1800 ppm CO2) for 2 h, 4 h, 24 h, and 7 d, respectively. At 2 h and 4 h, the expression of crh mRNA in gills increased with increasing CO2 concentration. Crh protein is expressed mainly in the lamellae cells. crhbp and crhr1 expression also increased significantly. However, at 2 h and 4 h, acidification caused little changes in these genes and Crh protein expression in the brain. At 7 d, Crh-positive cells were detected in the hypothalamus; moreover, Crh protein expression in the whole brain increased. It is suggested that CRH autocrine secretion in gills is responsible for local acid-base regulation rather than systemic mobilization after short-term acidification stress, which may help the rapid regulation of body damage caused by environmental stress.

Microbiome characterization and re-design by biologic agents for inflammatory bowel disease insights.

The therapeutic effect of inflammatory bowel disease has improved in the past decades, but most of patients cannot tolerate, do not respond to drugs, or relapse after treating with conventional therapy. Therefore, new and more effective treatment methods are still needed in treatment of IBD. In this review, we will discuss the relevant mechanisms and the latest research progress of biologics (anti-TNF treatments, interleukin inhibitors, integrin inhibitors, antisense oligonucleotide, and JAK inhibitors) for IBD, focus on the efficacy and safety of drugs for moderate-to-severe IBD, and summarize the clinical status and future development direction of biologics in IBD.

Ultrasensitive Gas Refractometer Using Capillary-Based Mach-Zehnder Interferometer.

In this paper, we report a capillary-based Mach-Zehnder (M-Z) interferometer that could be used for precise detection of variations in refractive indices of gaseous samples. The sensing mechanism is quite straightforward. Cladding and core modes of a capillary are simultaneously excited by coupling coherent laser beams to the capillary cladding and core, respectively. An interferogram would be generated as the light transmitted from the core interferes with the light transmitted from the cladding. Variations in the refractive index of the air filling the core lead to variations in the phase difference between the core and cladding modes, thus shifting the interference fringes. Using a photodiode together with a narrow slit, we could interrogate the fringe shifts. The resolution of the sensor was found to be ~5.7 × 10-8 RIU (refractive index unit), which is comparable to the highest resolution obtained by other interferometric sensors reported in previous studies. Finally, we also analyze the temperature cross sensitivity of the sensor. The main goal of this paper is to demonstrate that the ultra-sensitive sensing of gas refractive index could be realized by simply using a single capillary fiber rather than some complex fiber-optic devices such as photonic crystal fibers or other fiber-optic devices fabricated via tricky fiber processing techniques. This capillary sensor, while featuring an ultrahigh resolution, has many other advantages such as simple structure, ease of fabrication, straightforward sensing principle, and low cost.

Polycarbonate mPOF-Based Mach-Zehnder Interferometer for Temperature and Strain Measurement.

In this paper, an endlessly single mode microstructured polymer optical fiber (mPOF) in a Mach-Zehnder (M-Z) interferometer configuration is demonstrated for temperature and strain measurement. Because there is no commercial splicer applied for POF-silica optical fiber (SOF) connectorization, prior to the M-Z interferometric sensing, we introduce an imaging projecting method to align a polycarbonate mPOF to a SOF and then the splice is cured permanently using ultraviolet (UV) glue. A He-Ne laser beam at 632.8 nm coupled in a SOF is divided by a 1 × 2 fiber coupler to propagate in two fiber arms. A piece of mPOF is inserted in one arm for sensing implementation and the interference fringes are monitored by a camera. For non-annealed fiber, the temperature sensitivity is found to be 25.5 fringes/°C for increasing temperature and 20.6 fringes/°C for decreasing temperature. The converted sensitivity per unit length is 135.6 fringes/°C/m for increasing temperature, which is twice as much as the silica fiber, or 852.2 rad/°C/m (optical phase change versus fiber temperature), which is more than four times as much as that for the PMMA fiber. To solve the sensitivity disagreement, the fiber was annealed at 125 °C for 36 h. Just after the thermal treatment, the temperature measurement was conducted with sensitivities of 16.8 fringes/°C and 21.3 fringes/°C for increasing and decreasing process, respectively. One month after annealing, the linear response was improved showing a temperature sensitivity of ~20.7 fringes/°C in forward and reverse temperature measurement. For the strain measurement based on non-annealed fiber, the sensitivity was found to be ~1463 fringes/%ε showing repeatable linear response for forward and reverse strain. The fiber axial force sensitivity was calculated to be ~2886 fringes/N, showing a force measurement resolution of ~3.47 × 10-4 N. The sensing methodology adopted in this work shows several advantages, such as very low cost, high sensitivity, a straightforward sensing mechanism, and ease of fabrication.

[Correlation of aldo-keto reductases with prostate cancer and intervention with traditional Chinese medicine].

The androgen receptor signaling pathway is a key factor in the development and progression of prostate cancer. Aldo-keto reductases AKR1C1-AKR1C4 play an important role in the synthesis and metabolism of androgens in the body, and their expressions influence the androgen receptor signaling pathway and consequently the development and progression of prostate cancer. For the treatment of androgen-resistant prostate cancer, which cannot be cured currently, Chinese medicine and phytotherapy are receiving more and more attention for the mild, long-lasting and multi-target advantages of the small molecules of traditional Chinese medicine. This review summarizes the roles of aldo-keto reductases in the progression of prostate cancer and compares the anti-tumor activities of small molecules in Chinese medicine targeting aldo-keto reductases, hoping to provide a basis for the discovery of new targets for prostate cancer and the development of anti-tumor drugs.

CPP2-p16MIS treatment-induced colon carcinoma cell death in vitro and prolonged lifespan of tumor-bearing mice.

BACKGROUND: Cell-penetrating peptides (CPPs) are a research hotspot due to their noninvasive delivery ability. Among the identified CPPs, the TAT and R8 peptides have been preferentially applied to transduction into different cells. However, this process is nonselective among various cells. Recent research suggested that CPP2 could selectively penetrate human colorectal cancer (CRC) cells. METHODS: Using in vitro experiments, the mean fluorescence intensity of fluorescein isothiocyanate-labeled CPPs (CPPs-FITC) incubated with different cell lines was compared to corroborate the colon tumor targeting ability of CPP2. The targeting ability of CPP2 was determined in the same way in tumor-bearing mice. We synthesized antitumor peptides by fusing CPP2 to the minimal inhibitory sequence of p16 (p16MIS), which had the ability to restore the function of lost p16, the expression of which was absent in tumor cell lines of various origins. The antitumor effect of the combined peptide was tested in both CRC cell lines and tumor-bearing mice. RESULTS: In each CRC cell line, the mean fluorescence intensity of CPP2-FITC was higher than that of the TAT-FITC (p < 0.001) and R8-FITC (p < 0.001) groups. CPP2-p16MIS, the targeting carrier, showed a higher antitumor response in the in vitro cell research. CPP2-p16MIS showed a prolonged mean lifespan of tumor-bearing mice, further characterizing its role in specific tumor-targeting ability in vivo. Survival analysis showed that the mice treated with CPP2-p16MIS had significantly longer survival than the mice treated with phosphate-buffered saline (p < 0.05) or those treated with control peptides, including the CPP2 (p < 0.05) and p16MIS (p < 0.05) groups. CONCLUSION: CPP2 could more selectively penetrate CRC cells than TAT or R8 as well as effectively deliver the p16MIS to the tumor.

Microglial activation and pyroptosis induced by nano-TiO2 in marine medaka brain.

Recently, nano-titanium dioxide (nano-TiO2) has been widely distributed over surface water. However, there are few reports on its effects on the central nervous system of fish. In this study, we investigated whether nano-TiO2 enters the medaka brain after exposure and its effect on the brain. Marine medaka brains were examined after exposure to 0.01 g/L nano-TiO2 for 3, 10, and 20 d. Nano-TiO2-like particles were found in the telencephalon of treated fish. There was no obvious brain histopathological injury. The number of irregular mitochondria with absent cristae increased. Gene expression of the apoptosis-related genes, casp8, bcl2b, and bax, decreased significantly in the nano-TiO2 group at 3 d. In contrast, the pyroptosis-related genes, gsdmeb and casp1, and inflammation-related factor, il18, increased significantly. As an activated microglia marker, mRNA expression of cd68 increased significantly in the nano-TiO2 treated group. Moreover, CD68 protein expression also increased significantly at 10 d. Altogether, we show that nano-TiO2 can alter mitochondrial morphology in the telencephalon of medaka, leading to microglial activation and pyroptosis.

Versatile Fabrication of Biocompatible Antimicrobial Materials Enabled by Cationic Peptide Bundles.

Antimicrobial peptides (AMPs) are expected to be an alternative promising solution to the global public health problem of antibiotic resistance due to their unique antimicrobial mechanism. However, extensive efforts are still needed to improve the shortcomings of traditional AMPs, such as rapid proteolysis, hemolysis, slow response, toxicity, etc., by exploring AMP-based new antimicrobial strategies. Here, we develop cationic peptide bundles into novel antimicrobial architectures that can rapidly kill multiple types of bacteria including drug-resistant bacteria. Remarkably, cationic peptide bundles can be used as polymerization units to cross-link with other polymers through simple two-component polymerization to produce diverse antimicrobial materials. For the proof of concept, three materials were fabricated and investigated, including an antimicrobial hydrogel that can significantly accelerate the healing of infected wounds, a multifunctional antimicrobial bioadhesive that shows promise in antimicrobial coatings for medical devices, and a photo-cross-linked antimicrobial gelatin hydrogel with broad application potential. The integration of antimicrobial units into the materials' backbone endows their biocompatibility. Cationic peptide bundles not only represent a new antimicrobial strategy but also provide a versatile and promising processing method to create diversified, multifunctional, and biocompatible antimicrobial materials.

Exosome as potential biomarkers for gastrointestinal tumors.

INTRODUCTION: Exosomes are polyvesicles that are formed by invagination of intracellular lysosomal particles, and are released into the extracellular matrix after the fusion of polyvesicular outer membrane and cell membrane. In the body, immune response, antigen presentation, cell migration, cell differentiation and tumor invasion are closely related to tumorigenesis and tumor progression. This study aimed to conduct a meta-analysis for evaluating the clinicopathological, diagnostic and prognostic significance of exosomal expression in gastrointestinal tumors. METHODS: The original English articles were systematically searched in the online databases. The diagnostic accuracy, prognostic utility and clinicopathological correlation of gastrointestinal tumors were investigated. The quality assessment for studies of diagnostic accuracy II and Newcastle-Ottawa scale were used for quality evaluation, and the data was strictly extracted to judge the deviation of the study. RESULTS: A total of 14 studies with 1837 gastrointestinal tumor patients were included. The change in exosomal expression showed significant correlation with poor clinicopathological parameters (tumor diameter: combined P = .00024394; differentiation: combined P = 2.796e-08; lymphatic metastasis: P = 9.610e-07; distant metastasis: combined P = .00017326; pathological classification: combined P = .00875213; invasion depth: combined P = 3.504e-08) carcinoembryonic antigen (combined P = . 04458857) and tumor location (combined P = .00145983). The difference in the area under the curve between gastrointestinal tumor patients and healthy people showed an area under the curve of 0.89 (95%Cl 0.85-0.91) and heterogeneity of 0.59, 95% CI=[0.55-0.68]. The sensitivity was 0.88 (95%Cl 0.83 mi 0.91), the specificity was 0.72 (95%Cl 0.63 mi 0.80), and the diagnostic odds ratio was 18 (10-33). The results of survival analysis revealed that the abnormally expressed exosomes were significantly correlated with poor overall survival (hazard ratio =2.81, 95% CI: 2.02-3.93, P=0.013∗ 62.7%∗). CONCLUSION: The abnormally expressed exosomes might act as auxiliary biomarkers in diagnosing gastrointestinal tumors and demonstrated good prognostic significance in predicting the survival of patients with gastrointestinal tumors.

Overexpression of Rictor protein in colorectal cancer is correlated with tumor progression and prognosis.

In order to understand the clinical significance of rapamycin-insensitive companion of mammalian target of rapamycin (Rictor) in colorectal cancer (CRC), 62 CRC tissue samples excised during operations were evaluated by immunohistochemistry. Analysis of the association between the expression level of Rictor protein and clinicopathological parameters demonstrated that the expression level of Rictor in CRC tissues was significantly higher than that in paracarcinoma tissues (P<0.0001). In cellular experiments, this result was further confirmed by comparing differences in Rictor expression between the CRC cell lines HCT116, SW480 and LoVo, and the human normal liver cell line HL-7702. It was also noticed that the expression of Rictor was associated with Dukes stage, lymphatic metastasis and prognosis, as determined by χ2 test, Kaplan-Meier analysis and log-rank test. These results suggest that Rictor may be a novel target for the treatment and prognostic assessment of CRC patients in the future.

[Expression of Rictor and mTOR in colorectal cancer and their clinical significance].

OBJECTIVE: To explore the expression of Rictor and mTOR in the colorectal cancer and their clinical significance. METHODS: The expression levels of Rictor and mTOR in HCT116, SW480, LoVo and HCoEpiC cells were detected by indirect immunofluorescence and Western blotting. Sixty-two paraffin-embedded surgical specimens of colorectal cancer tissue and adjacent tissues were examined for Rictor expression using immunohistochemistry. The association of the expression levels of Rictor protein with the clinicopathologic features and the overall survival of the patients was analyzed. RESULTS: The expression level of Rictor was significantly higher in colorectal cancer tissues than in the adjacent tissues (P<0.05). The expression levels of Rictor and mTOR in the colon cancer cell lines were higher than those in human normal colon epithelial cell line HCoEpiC. The expression of Rictor was correlated with Dukes stage and lymphatic metastasis of the tumors but not with other clinicopathological parameter (P>0.05). Patients with Rictor expression had a lower overall survival rate than those without Rictor expression. CONCLUSION: Rictor overexpression is associated with the carcinogenesis and progression of colorectal cancer and can be an independent indicator for evaluating the prognosis of colorectal cancer patients.

Gap junction composed of connexin43 modulates 5­fluorouracil, oxaliplatin and irinotecan resistance on colorectal cancers.

Chemotherapy is one of the most commonly used therapeutic strategies for metastatic colon cancer. However, the development of resistance to chemotherapeutic agents limits their application in clinical use. The underlying mechanisms of this resistance development require further elucidation. The current study investigated the effects of connexin43 (Cx43) gap junctions on 5­fluorouracil (5­FU), oxaliplatin and irinotecan in colon cancer cells. Three different methods were used to manipulate Cx43 gap junction function: i) Cell culture at different densities; ii) pretreatment with a Cx43 specific inhibitor or enhancer; and iii) Cx43 gene knock­down. Results indicated that the cell toxicity of 5­FU, oxaliplatin and irinotecan was cell density­dependent, which was mediated by gap junctions. Downregulation of Cx43 gap junction functioning attenuated 5­FU, oxaliplatin and irinotecan toxicity in colon cancer cells, which was increased in cells treated with a Cx43 gap junction function enhancer. Thus, the results of the present study suggest that resistance to 5­FU, oxaliplatin and irinotecan in colon cancer cells was relative to Cx43 expression loss as cancer developed, which may indicate a novel basis for therapeutic strategy development to combat drug resistance in numerous cell types, in addition to colon cancer cells.

[Effects of the lipid-lowering drugs on endothelial hyperplasia in inferior vena cava grafts in dogs].

OBJECTIVE: To investigate the effects of the lipid-lowering drugs in alleviating endothelial hyperplasia in the inferior vena cava (IVC) grafts in dogs. METHODS: The Dacron grafts seeded with autologous venous fragments were implanted into the IVC of 20 dogs, including 12 dogs receiving oral lipid-lowering drugs serving as the treatment group and the other 8 without medication as the control group. The levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-ch) and high-density lipoprotein cholesterol (HDL-ch) in the serum were measured regularly, and all the grafts harvested to measure the thickness of the endothelium. RESULTS: The total patency rate of the IVC were higher in the treatment group (75%) than in the control group (37.5%), and new endothelial lining was formed two weeks after the operation. Compared with the control group, the endothelial thickness of the grafts at the proximal (P<0.01), middle (P<0.05) and distal segments (P<0.05) of the IVC were all smaller in the treatment group, which also had lower serum LDL-ch and TC levels (both P<0.05) but with comparable HDL-ch levels (P>0.05). CONCLUSION: Administration of lipid-lowering drugs may reduce the level of serum LDL-ch and TC and the endothelial thickness of the grafts to improve the patency rate of the vessels.

[Application of three-dimensional high-definition laparoscope in laparoscopic radical resection of gastric cancer].

OBJECTIVE: To investigate the clinical value of three-dimensional (3D) high-definition (HD) laparoscope in laparoscopic radical resection of gastric cancer. METHODS: From January to December, 2013, 40 patients underwent radical resection of gastric cancer with 3D HD laparoscopy (3D group) and another 40 patients received 2D HD laparoscopy (2D group). The duration of surgery, intra-operative blood loss, learning curve, and costs during hospitalization were compared between the two groups. RESULTS: The average operation duration of 3D group was 2.8=0.6 h, significantly shorter that in the 2D group (3.2=0.8 h, P<0.05); the intraoperative blood loss in the 3D group was significantly less than that in the 2D group (110=18 ml vs 120=21 ml, P>0.05). The mean hospitalization cost was 75 000=16 000 RMB Yuan in 3D group, similar to significantly lower than that of 71 000=13 000 RMB Yuan in 2D group (P>0.05). CONCLUSION: 3D HD laparoscopy can provide three-dimensional vision and better sense of depth to facilitate precise operation and shorten the operation time. The high-definition 3D vision also allows surgeons to quickly improve surgical skills and shorten the learning curve.

Interference of Skp2 effectively inhibits the development and metastasis of colon carcinoma.

Colon cancer is a common type of malignancy in the digestive system. The aim of the present study was to investigate the role of S-phase kinase-associated protein 2 (Skp2) in colon carcinoma and to identify whether depletion of Skp2 by Skp2­RNA interference (RNAi) attenuates the proliferation and migration of colon carcinoma. Three pairs of small interfering (si)RNA were designed based on the Skp2 gene sequence and the most effective one was used to silence the Skp2 gene in SW620 cells. Subsequent to the interference, quantitative polymerase chain reaction and western blot analysis were used for detecting the expression of Skp-2 mRNA and protein, respectively. The data demonstrated that the Skp2­siRNA effectively inhibited proliferation (P<0.01), increased the levels of apoptosis and induced G0/G1 phase arrest of the SW620 cells (P<0.01). Transfection of the Skp2 siRNA into SW620 cells effectively reduced Skp2 protein levels, while p27 protein levels increased. In the in vivo experiments, a lentiviral vector of the Skp2-RNAi transfected into SW620 cells markedly inhibited Skp2 expression, as detected by immunohistochemical analysis of nude mice. Additionally, tumorigenicity experiments showed that inhibition of Skp2 significantly increased the survival rate of nude mice. Thus, the in vitro and in vivo results demonstrated that interference of Skp2 expression significantly inhibited the proliferation and induced the apoptosis of SW620 cells. This suggests that Skp2 protein has an important role in the progression of colon cancer. Therefore, Skp2 may enable the early diagnosis of colon cancer and provide new insights into molecular targets for cancer therapy.

Alpinetin attenuates inflammatory responses by interfering toll-like receptor 4/nuclear factor kappa B signaling pathway in lipopolysaccharide-induced mastitis in mice.

Alpinetin, a novel plant flavonoid derived from Alpinia katsumadai Hayata, has been reported to exhibit anti-inflammatory properties. However, the effect of alpinetin on mastitis has not been investigated. The aim of this study was to investigate the protective effect of alpinetin against lipopolysaccharide (LPS)-induced mastitis and to clarify the possible mechanism. In the present study, primary mouse mammary epithelial cells and an LPS-induced mouse mastitis model were used to investigate the effect of alpinetin on mastitis and the possible mechanism. In vivo, we observed that alpinetin significantly attenuated the infiltration of neutrophilic granulocytes, and the activation of myeloperoxidase; down-regulated the level of pro-inflammatory cytokines, including TNF-α, IL-1ß and IL-6; inhibited the phosphorylation of IκB-α, NF-κB p65 and the expression of TLR4, caused by LPS. In vitro, we also observed that alpinetin inhibited the expression of TLR4 and the production of TNF-α, IL-1ß and IL-6 in LPS-stimulated primary mouse mammary epithelial cells. However, alpinetin could not inhibit the production of IL-1ß and IL-6 in TNF-α-stimulated primary mouse mammary epithelial cells. In conclusion, our results suggest that the anti-inflammatory effects of alpinetin against LPS-induced mastitis may be due to its ability to inhibit TLR4-mediated NF-κB signaling pathways. Alpinetin may be a promising potential therapeutic reagent for mastitis treatment.

[Selective cytotoxic effect of lentivirus-mediated double suicide gene transfer on human gastric adneocarcinoma cells].

OBJECTIVE: To study the selective cytotoxic effect of lentivirus-mediated double suicide gene (CD/TK) against human gastric carcinoma cells SGC-7901 in vitro. METHODS: SGC-7901 cells were infected with FGW-KDRP-CD/TK vector and the infection efficiency was observed under a fluorescence microscope. The morphological changes of the infected cells were observed by Giemsa staining. Flow cytometry (FCM) was employed for cell cycle analysis, and the expression of CD/TK was detected by RT-PCR. The infected cells were then treated with the prodrugs ganciclovir (GCV) and/or 5-fluorocytosine (5-FC) at different concentrations, and the cytotoxic effects were evaluated using MTT method. RESULTS: The infection efficiency of the lentiviral vector in SGC-7901 cells increased with the titer of the virus, which produced no significant effect on the cancer cell morphology in vitro or on the percentages of G0-G1, G2-M and S phase cells (P>0.05). RT-PCR demonstrated the expression of CD/TK gene in SGC-7901 cells infected by FGW-KDRP-CD/TK. The infected cells were highly sensitive to the prodrugs with a dose-dependent cytotoxic effect within a specific concentration range of the drugs, whereas the non-infected cells were not sensitive to the prodrugs. Combined use of the two prodrugs produced an obviously stronger inhibitory effect than either of the them (P<0.05). When combined, GCV and 5-FC at the concentration of 0.1+40, 1+80, 10+160, and 100+320 mg/L demonstrated a synergetic effect with a CDI<1. CONCLUSION: Lentivirus-mediated CD/TK fusion gene system can selectively kill gastric cancer cells, and the two prodrugs show a synergistic cytotoxic effect.

[Preliminary effect of VEGF promoter-driven recombinant adenovirus containing double suicide genes on apoptosis of human gastric carcinoma cells].

OBJECTIVE: To study the effect of the adenovirus containing CD/TK fusion gene controlled by the human vascular endothelial growth factor (VEGF) promoter on apoptosis of human gastric carcinoma cells SGC-7901. METHODS: VEGF-expressing SGC-7901 cells were infected by the recombinant adenovirus Ad-VEGFP-CD/TK, and the infection efficiencies were observed with fluorescence microscopy. The toxic effect and intracellular calcium concentration induced by 5-fluorocytosine (5-FC) and ganciclovic (GCV) were determined by light microscopy, electron microscopy and flow cytometry. RESULTS: The transfection efficiency of the recombinant adenovirus in SGC-7901 cells increased with the viral titer. At the multiplicity of infection (MOI) of 100, 5-FC and GCV could induce apoptosis of SGC-7901 cells within a given dose range in a dose- and time-dependent manner, and apoptotic changes of the cells were observed with electron microscopy. Apoptotic peak was also detected by flow cytometry. Cell cycle analysis revealed increased cell percentage in G(0)-G(1) phase and decreased percentage of cells in G(2)-M and S phases in response to treatment with the pro-drugs, which also induced marked elevation of intracellular calcium concentration in the infected cells. CONCLUSIONS: CD/TK fusion gene system driven by VECF promoter selectively induces apoptosis of VEGF-expressing SGC-7901 cells, the action of which is probably mediated by intracellular calcium variation.

[Targeted killing of colorectal tumor cells by lentiviral constructs containing CD/TK suicide genes and KDR promoter].

OBJECTIVE: To investigate the selective killing of colorectal tumor cells by lentivirus-mediated double suicide gene under the regulation of KDR promoter. METHODS: 293T packaging cells were transfected with the plasmid FGW-KDRP-CD/TK to obtain the infectious viruses. KDR-expressing LoVo cells and LS174T cells that did not produce KDR were transfected with the recombinant virus, and the transfection efficiency was evaluated by the fluorecence microscope. RT-PCR was employed to examine the expression of CDglyTK. After treatment of the cells with 5-FC and GCV, the killing effects on the two cell lines were evaluated. RESULTS: The recombinant construct showed similar infection rate of the two cell lines. RT-PCR demonstrated that CDglyTK gene was expressed only in LoVo cells infected with FGW-KDRP-CD/TK but not in LS147T cells, and the sensitivity of the two cell lines to the prodrugs was significantly different (P<0.001). The killing effect of the double suicide gene was much stronger than that of single suicide gene administered (P<0.001). CONCLUSION: The double suicide gene driven by KDR promoter has specific killing effect on the KDR-expressing colorectal tumor cells.