[Value of double contrast-enhanced ultrasound QontraXt three-dimensional pseudocolor quantitative analysis to therapeutic effect evaluation of preoperative neoadjuvant chemotherapy in advanced gastric cancer patients].

Objective: To investigate the value of tumor perfusion parameter measured by using double contrast-enhanced ultrasound (DCEUS) QontraXt three-dimensional pseudocolor quantitative analysis to the therapeutic effect evaluation of preoperative neoadjuvant chemotherapy (NAC) in advanced gastric cancer (AGC) patients. Methods: Eighty-nine AGC patients underwent 3 cycles of preoperative NAC (XELOX) followed by complete resection of lesion. The DCEUS QontraXt three-dimensional pseudocolor was performed one or two weeks before the NAC and operation were applied, respectively. The peak enhancement (PE), time to peak (TP), sharpness of the bolus (ß) and area under the enhancement curve (AUC) of primary gastric tumor were measured by QontraXt three-dimensional pseudocolor quantitative analysis. These DCEUS parameters between respond and non-respond groups before and after NAC therapy were compared. The prediction accuracy of DCEUS to the therapeutic effect evaluation of preoperative NAC was determined by the receive operating characteristic (ROC) curves. Results: Among 89 AGC patients, 52 patients responded to NAC therapy, while 37 patients resisted to NAC therapy. Twelve cases in respond group and 26 cases in non-respond group were mucinous carcinoma. Forty cases in respond group and 11 cases in non-respond group were non-mucinous carcinoma (P<0.05). In responder group, the PE and TP before NAC were (53.7±9.3)% and (14 521±2 667) ms, and (32.2±5.5)% and (17 235±1 898) ms after NAC. The ratio of changes of PE (ΔPE) and TP (ΔTP) were 0.43±0.17 and 0.36±0.14, respectively. In non-respond group, the PE and TP before NAC were (54.4±7.2)% and (13 869±3 247) ms, and (45.3±6.1)% and (15 127±1 423) ms after NAC therapy. The ratio of ΔPE and ΔTP were 0.24±0.20 and 0.22±0.12. The PE and TP after NAC, the ratio of ΔPE and ΔTP were significant different among these two groups (all of P<0.05). The ROC curves showed that the ratio of ΔPE in assessing the respond of gastric cancer patients to NAC was superior compared to other parameters (AUC=0.784, P=0.004). The optimal cut-off value of the ratio of ΔPE was 24% and its sensitivity and specificity to the therapeutic effect evaluation of NAC in gastric cancer were 82.7% and 64.9%. Conclusion: DCEUS QontraXt three-dimensional pseudocolor quantitative analysis might be a novel, noninvasive and reliable method to evaluate the therapeutic effect of preoperative NAC in AGC patients.

[Comparison of aortic annular diameter defined by different measurement mordalities before transcatheter aortic valve implantation].

Objective: To compare aortic annular diameter measured by transthoracic echocardiography (TTE), transesophageal echocardiography (TEE), and multislice computed tomography (MSCT) in patients with severe aortic stenosis, and to evaluate the impact on selection of prosthetic valve type in transcatheter aortic valve implantation (TAVI). Methods: Clinical data of 138 patients with severe aortic stenosis referred for TAVI between January 2014 and June 2016 in our hospital were retrospectively analyzed.The difference of aortic annular diameter measured by TTE, TEE, and MSCT were compared.TTE was performed after TAVI to evaluate the accuracy of measurement before TAVI. Results: (1) Aortic annular diameter was (23.37±2.22) mm by TTE and (23.52±1.70) mm by TEE (P=0.12). Pearson correlation analysis showed that aortic annular diameter measured by TTE was correlated to that measured by TEE (r=0.87, P<0.05). (2)The long-axis diameter and the short-axis diameter measured by MSCT multiplanar reconstruction were significantly different ((27.86±2.87) mm vs. (21.91±2.53) mm, P<0.05). There was no significant difference between the mean of the long- and short-axis diameters and the diameter derived from cross-sectional area by MSCT ((24.92±2.38) mm vs. (24.84±2.25) mm, P=0.08). However, the diameter derived from the circumference ((25.35±2.34) mm) was significantly larger than the mean of the long- and short-axis diameters and the diameter derived from cross-sectional area by MSCT, and the difference were (0.43±0.62) mm and (0.51±0.62) mm respectively(both P<0.05). (3) Measurements of the aortic annulus diameter by MSCT including the mean of the long- and short-axis diameters, diameter derived from cross-sectional area, and diameter derived from the circumference were larger than the measurement by TTE and TTE (all P<0.05). (4) Implantation was successful in all patients.Moderate to severe paravalvular leakage was detected in 3 patients at 7 days post TAVI, and 1 patient developed severe prosthetic valve restenosis at 6 months post TAVI and received valve-in-valve implantation. Conclusions: In severe aortic stenosis patients referred for TAVI, the aortic annular diameter derived from TTE and TEE measurements are smaller than that from MSCT.In the absence of a gold standard, selection of prosthetic valve type in TAVI procedure should rely on comprehensive considerations, which is of importance to get good clinical results for severe aortic stenosis patients underwent TAVI.

Isolation of biosurfactant producers, optimization and properties of biosurfactant produced by Acinetobacter sp. from petroleum-contaminated soil.

AIMS: To screen and identify biosurfactant producers from petroleum-contaminated soil; to use response surface methodology (RSM) for medium optimization to enhance biosurfactant production; and to study the properties of the newly obtained biosurfactant towards pH, temperature and salinity. METHODS AND RESULTS: We successfully isolated three biosurfactant producers from petroleum-contaminated soil and identified them through 16S rRNA sequence analysis, which exhibit the highest similarities to Acinetobacter beijerinckii (100%), Kocuria marina (99%) and Kineococcus marinus (99%), respectively. A quadratic response model was constructed through RSM designs, leading to a 57·5% increase of the growth-associated biosurfactant production by Acinetobacter sp. YC-X 2 with an optimized medium: beef extract 3·12 g l(-1) ; peptone 20·87 g l(-1) ; NaCl 1·04 g l(-1); and n-hexadecane 1·86 g l(-1). Biosurfactant produced by Acinetobacter sp. YC-X 2 retained its properties during exposure to a wide range of pH values (5-11), high temperatures (up to 121°C) and high salinities [up to 18% (w/v) Na(+) and Ca(2+) ], which was more sensitive to Ca(2+) than Na(+). CONCLUSIONS: Two novel biosurfactant producers were isolated from petroleum-contaminated soil. Biosurfactant from Acinetobacter sp. YC-X 2 has good properties to a wide range of pH, high temperature and high salinity, and its production was optimized successfully through RSM. SIGNIFICANCE AND IMPACT OF THE STUDY: The fact, an increasing demand of high-quality surfactants and the lack of cost-competitive bioprocesses of biosurfactants for commercial utilization, motivates researchers to develop cost-effective strategies for biosurfactant production through isolating new biosurfactant producers with special surface-active properties and optimizing their cultural conditions. Two novel biosurfactant producers in this study will widen our knowledge about this kind of micro-organism. This work is the first application of RSM designs for cultural optimization of biosurfactant produced by Acinetobacter genus and the first report that biosurfactant may be more sensitive to Ca(2+) than Na(+) .

Reappraisal of the importance of mutations in the NS5A-PKR-binding domain of hepatitis C-1b virus in the era of optimally individualized therapy.

Past studies have reported that mutations in the protein kinase R-binding domain (PKRBD) sequences of hepatitis C virus (HCV) NS5A proteins are correlated with response to fixed-duration interferon (IFN)-based therapy in patients infected with HCV-1b. In this study, we investigated whether the substitutions in PKRBD, including the IFN sensitivity-determining region (ISDR) and 26 additional downstream amino acids from ISDR, will have effects upon patients infected with chronic HCV-1b in the era of individualized therapy with peginterferon and ribavirin. Thirty-seven patients were treated with optimally tailored therapy guided by baseline viral load combined with rapid and early virological responses while 23 patients were treated without guidance and/or assigned suboptimal treatment duration. The amino acid sequences of the PKRBD were determined by PCR and sequencing. The overall sustained virological response (SVR) rate of patients who received optimally individualized therapy was 78.4%, which was better than the SVR rate of patients who received suboptimal therapy (47.8%, P = 0.015). Multivariate analysis showed that optimally individualized therapy (P = 0.019) and 80/80/80 adherence (P = 0.006) were independent favourable predictors of SVR in the entire cohort. Further sub-analysis of the predictive factors of SVR in patients treated with optimally individualized therapy showed that mutations in the 26-amino acid downstream from the ISDR (P = 0.024) were the only independent predictor of SVR. We concluded that mutations in 26-amino acid downstream portion from the ISDR remained a prognosticator of SVR in the era of optimally tailored therapy.

A novel glioblastoma cancer gene therapy using AAV-mediated long-term expression of human TERT C-terminal polypeptide.

Glioblastoma multiforme is the most aggressive form of human brain tumor, which has no effective cure. Previously, we have demonstrated that overexpression of the C-terminal fragment of the human telomerase reverse transcriptase (hTERTC27) inhibits the growth and tumorigenicity of human cervical cancer HeLa cells. In this study, the therapeutic effect and molecular mechanisms of hTERTC27-mediated cancer gene therapy were further explored in vivo in established human glioblastoma xenografts in nude mice. We showed that intratumoral injection of adeno-associated virus carrying hTERTC27 (rAAV-hTERTC27) is highly effective in reducing the growth of the subcutaneously transplanted glioblastoma tumors. Histological analyses showed that rAAV-hTERTC27 treatment leads to profound necrosis, apoptosis, infiltration of polymorphonuclear neutrophils and reduced microvessel density in the tumor samples. To study the molecular mechanism of rAAV-hTERTC27-mediated antitumor effects, we analyzed the global gene expression profiles of the rAAV-hTERTC27-treated tumor tissues and cell line as compared with that of the control rAAV-green fluorescent protein-treated samples by DNA microarray. Our results suggest that hTERTC27 exerts its effect through complex mechanisms, which involve genes regulating apoptosis, cell adhesion, cell cycle, immune responses, metabolism, signal transduction, transport, transcription and telomere maintenance.

Effects of alteration in the light cycle on outer segment shedding in the fetal retina.

Shedding of outer segment discs and their phagocytosis by retinal pigment epithelial cells (RPE) in the guinea pig starts to occur during in utero development. The number of phagosomes in the fetal RPE varies in a cyclical manner somewhat analogous to that observed in mature animals. The present study addresses the question of whether the cycles in fetus and mother are independent or linked. Pregnant guinea pigs were subjected to 10 days of alteration of their lighting schedules beginning on the 46th day of gestation. Dams and fetuses were sacrificed on day 56, and large phagosomes in the RPE of each were counted. A 10-hour phase advance was employed, ie, from 0800 (lights on): 2000 (light off) in controls, to 2200 (on): 1000 (off) in experimental animals. Counts of large phagosomes in the posterior retina were compared in both control and experimental nulliparous, gravid, and fetal animals. A significant burst of shedding occurred within one hour after light onset in the experimental groups comprised of either nulliparous or pregnant animals. The latter also showed a smaller rise in phagosome numbers at the original time of onset of illumination. In contrast, no shift in the time of maximum shedding activity was observed in fetuses in the experimental regime. Their activity was similar to that present in the original 0800:2000 cycle. Whereas maternal RPE shedding cycles can be reentrained in a 10-day period, those in the 46- to 56-day fetal retina are refractory to significant change. Based on this finding, it is likely that the shedding cycles in the mother and fetus at this age are under independent control, and, therefore, that shedding in the fetus is not mediated by maternal borne factors.

Phagocytosis in the fetal pigment epithelium: evidence for cyclic activity.

Photoreceptor cells in guinea pig retinas begin to shed their outer segments while developing in utero. To ascertain whether phagocytosis of outer segments in the fetus is a cyclical event like that of mature animals, large phagosomes in fetal retinal pigment epithelial cells were counted at representative times during a 24 hr period. These data were then compared with similar data obtained from the pregnant dams. Gravid animals and fetuses both showed a burst of shedding after the onset of illumination. Shedding was well initiated in the pregnant animals 30 min after the onset of illumination but was delayed for 2 to 3 hr in their fetuses. Thereafter the two patterns of phagocytosis showed substantial differences. Congruent cyclical patterns could be discerned, however, by graphing fetal counts with an advance of 10 hr on the time scale relative to that of the gravid animals. The results presented here represent the first indication of a cyclical pattern of phagocytosis in a population of fetuses during retinal development.

The morphology and function of healing cat corneal endothelium.

We mechanically damaged the entire corneal endothelium of one eye of each of ten cats and then examined both eyes by fluorophotometry and specular microscopy for 5 months. Six weeks after damage, when the corneas had cleared sufficiently to make accurate measurements, the mean endothelial permeability to carboxyfluorescein was increased 11% (P = 0.02) and the mean central corneal thickness was increased 11% (P = 0.05) in the damaged eyes. The mean endothelial pump rate was decreased 29% (P = 0.05), indicating that the increase in permeability was insufficient to explain the increase in thickness. The permeability returned to normal by 3 months and the pump rate by 5 months. Six weeks after damage, the mean endothelial cell size was increased 89% (P less than 0.01), the mean coefficient of variation of cell size was increased 200% (P less than 0.01), and the mean percentage of hexagonal cells was decreased 34% (P less than 0.01). By 5 months, the mean cell size had changed very little, and none of the three morphologic measurements had returned to normal. As in rabbits, the endothelial barrier in cats recovers before the pump after wounding. Unlike in rabbits, functional recovery in cats requires at least several months. Such prolonged functional recovery after endothelial trauma might also be expected in humans who, like cats and unlike rabbits, have little capacity for endothelial mitosis during healing.

Long-term observation of morphologic and functional features of cat corneal endothelium after wounding.

PURPOSE: (1) To test the hypothesis that corneas with enlarged endothelial cells (and thus less intercellular space) have decreased endothelial permeability to small polar solutes. (2) To measure corneal endothelial ouabain binding (Na+/K+ ATPase "pump site" density) and Descemet's membrane production after endothelial wounding. METHODS: Bilateral specular microscopy and anterior segment fluorophotometry were performed at 2-month intervals for 1 year in ten cats after mechanically damaging the corneal endothelium in one eye of each. The measurements were repeated at 2 years in four cats and at 3 years in two cats. Eighteen months after wounding, endothelial ouabain binding was measured in both eyes of six cats. Transmission electron micrographs of Descemet's membrane were analyzed in both eyes of six cats at 18 months, two cats at 2 years, and two cats at 3 years after wounding. RESULTS: From 6 to 12 months after wounding, the endothelial permeability to carboxyfluorescein was significantly decreased (P < 0.05), and the mean endothelial cell size was significantly increased (P < 0.001) in the damaged eyes. The enlarged endothelial cells persisted in the few cats observed 2 and 3 years after wounding. There was no significant difference in endothelial ouabain binding between the damaged and control corneas in six cats tested 18 months after wounding. On subsequent histologic examination, a layer of abnormal Descemet's membrane was present in all ten wounded eyes, with additional normal Descemet's membrane posterior to it, between the abnormal layer and the endothelial cells. CONCLUSIONS: The results are consistent with the hypothesis that corneal endothelial permeability to small polar solutes varies directly with the amount of intercellular space available for diffusion across the monolayer. The results also confirm clinical reports of decreased endothelial permeability in corneas with enlarged endothelial cells. In histopathologic specimens, a layer of abnormal Descemet's membrane can be a historical marker for a period of endothelial damage and corneal decompensation.

Conopeptides from Conus striatus and Conus textile by cDNA cloning.

Conopeptide content in Conus textile and Conus striatus venoms were examined by polymerase chain reaction amplification of alpha-conopeptide cDNA and rapid amplification of 3' cDNA ends of O-superfamily conopeptide cDNA. Two new alpha-conopeptide sequences and six new O-superfamily conopeptide sequences from C. textile, four new O-superfamily conopeptide sequences, and four previously biochemically characterized conopeptide sequences from C. striatus were identified. The results suggest that this cDNA method is rapid and requires less material for the study of conopeptides.

Effectiveness of postoperative adjuvant therapy in improving reproductive outcome of endometriosis-associated infertility.

BACKGROUND AND PURPOSE: Treatment of endometriosis-associated infertility has not yet become standardized. Various protocols including surgical treatment, medical therapy, and a combination of both have been suggested but their use remains controversial. The objective of the present study was to determine whether postoperative adjuvant therapy for endometriosis is effective in improving reproductive outcome. METHODS: Medical records of infertile patients with newly diagnosed endometriosis treated in a university teaching hospital during a 50-month period were reviewed. After exclusion of patients with other major infertility factors, a total of 209 patients were included in the retrospective analysis. These patients were divided into those receiving (n = 78) or not receiving (n = 131) peri- or postoperative adjuvant medical therapy. The adjuvant therapies included danazol (n = 62), gonadotropin releasing hormone analogues (n = 11), progestins (n = 3), oral contraceptives (n = 1), and mixed treatment (n = 1). RESULTS: The pregnancy rate was lower in those receiving adjuvant therapy, although this result was not significant (32.1% vs 45.8%; p = 0.05). When patients using postoperative danazol therapy were considered alone, the pregnancy rate in patients receiving adjuvant therapy was significantly lower than that in patients not receiving it (p = 0.047). When the stage of endometriosis was considered, the pregnancy rate in patients receiving adjuvant therapy was again lower than in those not receiving it in patients with minimal or mild endometriosis (42.9% vs 60%; p = 0.043). However, in patients with moderate or severe endometriosis, the pregnancy rate was not different in the two groups (31% vs 36%; p = 0.56). Postoperative assisted reproductive techniques (ART) including controlled ovarian hyperstimulation/intrauterine insemination (COH/IUI) and in vitro fertilization (IVF) were effective in improving the pregnancy rates for all patients (53.9% with ART vs 33.1% without; p = 0.003) and for patients with advanced endometriosis (47.7% with ART vs 27.2% without; p = 0.016). CONCLUSIONS: Our results suggest that postoperative adjuvant therapy is ineffective in improving reproductive outcome in patients with either early (minimal or mild) or advanced (moderate and severe) endometriosis. This finding suggests that if fertility is the goal of treatment, adjuvant therapy may be unnecessary after surgery. In contrast, our data suggest that empirical ART, including COH/IUI or IVF, may be a better alternative to improve the pregnancy outcome after surgery.

The feasible sequential control strategy of treating high strength organic nitrogen wastewater with sequencing batch biofilm reactor.

The bio-kinetics and feasible sequential control strategy of treating high strength organic carbon and nitrogen wastewater were investigated by conducting the ABS manufacturing wastewater in a series of Sequencing Batch Biofilm Reactors (SBBRs). The on-line ORP, pH, and DO monitoring parameters were applied to identify the feature-points when ammonification, nitrification, and denitrification ends. The carbonaceous matter removal kinetics in the anaerobic and aerobic reaction stages can be expressed by the Michaelis-Menten equation. High efficiency of organic carbon removal and organic nitrogen ammonification in the anaerobic stage can eliminate the substrate competition and activation inhibition to nitrifying organisms in the following aerobic stage. In the sequencing nitrogen removal processes, the producing time and system ORP values of these feature-points have good function relationships with the influent COD loading rates of SBBR, which can be integrated into a set-point (set-time and set-ORP) sequential control strategy of nitrogen removal. The automatic control operation results revealed ORP was one of the major control parameters of the sequencing nitrogen removal process in SBBR system and high overall removal efficiency were obtained.

[Bovine alpha-sl-casein gene sequences direct expression of a variant of human tissue plasminogen activator in the milk of transgenic mice].

The fusion gene containing the promoter of casein gene, LAtPA minigene and 3' flanking sequences of casein gene was introduced into the fertilized eggs of mice by microinjection. Five positive transgenic mice were obtained. The concentration of LAtPA in the milk of one female transgenic mouse is 0.18 microgram/ml. This result showed that the LAtPA minigene could correctly express the bioactive LAtPA in the milk of the transgenic mouse under the control of the regulatory elements of the casein gene.

Recombinant bivalent live vaccines against neonatal colibacillosis in piglets.

The genes of colonization factor K88 and avirulent heat-labile enterotoxin LT A-B+ of enterotoxigenic E. coli (ETEC) have been ligated, and a vaccine strain containing recombinant plasmid that efficiently expresses two antigens has been obtained. Using the activity of beta-galactosidase as a selection marker instead of drug resistance, another bivalent vaccine strain with the same expression level has also been constructed. The vaccine strains have no toxicity and do not cause any adverse reactions. In challenge study and field trials, a high degree of protection from colibacillosis was afforded to piglets when their dams were immunized orally or parenterally. Practice of bivalent live vaccines including colonization factor and enterotoxin antigens and without antibiotic resistance gene shows effective.

Performance of risk estimation for hepatocellular carcinoma in chronic hepatitis B (REACH-B) score in classifying treatment eligibility under 2012 Asian Pacific Association for the Study of the Liver (APASL) guideline for chronic hepatitis B patients.

BACKGROUND: REACH-B [Risk Estimation for Hepatocellular Carcinoma (HCC) in Chronic Hepatitis B] scoring system was developed to predict the risk of HCC in noncirrhotic chronic hepatitis B (CHB) patients. AIM: To evaluate the discriminatory performance of REACH-B scoring system in classifying anti-viral treatment eligibility of CHB patients according to the 2012 Asian Pacific Association for the Study of the Liver (APASL) treatment guideline. METHODS: A total of 904 noncirrhotic CHB were enrolled. Patients' age, gender, liver biochemistry, HBeAg status and HBV DNA levels were recorded. RESULTS: The minimum REACH-B risk score for patients to be eligible for anti-viral treatment was 7 for HBeAg-positive and 6 for HBeAg-negative patients. Among them, increasing REACH-B score was not significantly associated with eligibility for treatment [adjusted odds ratio (OR): 1.210, 95% confidence interval (CI): 0.979-1.494, P = 0.078] in HBeAg-positive patients, as shown by logistic regression analysis after adjusting for gender. In HBeAg-negative patients, REACH-B score significantly predicted the treatment eligibility (adjusted OR: 1.783, 95% CI: 1.607-1.979, P < 0.001). Discriminatory ability of REACH-B score to classify eligibility was poor for HBeAg-positive patients ≥40 years [area under receiver operating characteristic (AUC): 0.664, 95% CI: 0.533-0.795], but good/excellent for HBeAg-positive patients <40 years (AUC: 0.903; 95% CI: 0.841-0.964), HBeAg-negative patients ≥45 years (AUC: 0.883; 95% CI: 0.848-0.917) and HBeAg-negative patients <45 years (AUC: 0.907; 95% CI: 0.874-0.940). CONCLUSION: The discriminatory performance of the REACH-B scoring system in classifying anti-viral treatment eligibility based on the 2012 APASL guideline was good/excellent, except for ≥40 years old HBeAg-positive patients.

Enhancement of CPT-11 antitumor activity by adenovirus-mediated expression of ß-glucuronidase in tumors.

CPT-11 is a clinically important prodrug that requires conversion into the active metabolite SN-38, a potent topoisomerase I poison, for antitumor activity. However, SN-38 is rapidly metabolized to the inactive SN-38 glucuronide (SN-38G) in the liver, which reduces the amount of SN-38 available for killing cancer cells. Here, we investigated if local expression of ß-glucuronidase (ßG) on cancer cells to catalytically convert SN38G to SN38 could enhance the antitumor activity of CPT-11. ßG was tethered on the plasma membrane of three different human cancer cell lines: human colon carcinoma (LS174T), lung adenocarcinoma (CL1-5) and bladder carcinoma (EJ). Surface ß-glucuronidase-expressing cells were 20 to 80-fold more sensitive to SN-38G than the parental cells. Intravenous CPT-11 produced significantly greater suppression of CL1-5 and LS174 T tumors that expressed ßG as compared with unmodified tumors. Furthermore, an adenoviral vector expressing membrane-tethered ßG (Ad.ßG) increased the sensitivity of cancer cells to SN-38G even at multiplicity of infections as low as 0.16, indicating bystander killing of non-transduced cancer cells. Importantly, intratumoral injection of Ad.ßG significantly enhanced the in vivo antitumor activity of CPT-11 as compared with treatment with CPT-11 or Ad vectors alone. This study shows that Ad.ßG has potential to boost the therapeutic index of CPT-11.

Development of recombinant adeno-associated virus and adenovirus cocktail system for efficient hTERTC27 polypeptide-mediated cancer gene therapy.

The low in vivo transduction efficiency of recombinant adeno-associated virus (rAAV) and the undesirably strong immunogenicity of adenovirus (rAdv) have limited their clinical utilization in cancer gene therapy. We have previously demonstrated that intratumoral injection of rAAV expressing a C-terminal polypeptide of human telomerase reverse transcriptase (rAAV-hTERTC27) effectively inhibits the growth of glioblastoma xenografts in nude mice. To further improve its efficacy, we combined rAAV-hTERTC27 with rAdv and investigated the efficiency of the cocktail vectors in vivo. At a nontherapeutic dose (1 x 10(8) plaque-forming units (PFUs)), rAdv-null and rAdv-hTERTC27 were equipotent in enhancing the therapeutic efficacy of rAAV-hTERTC27 (1.5 x 10(11) v.g.), and complete tumor regression was achieved in 25% of the treated animals. Importantly, the combination of rAAV-hTERTC27 and a therapeutic dose (2.5 x 10(9) PFU) of rAdv-hTERTC27 significantly augmented the therapeutic effects and led to a 38% complete tumor regression rate. In vivo optical imaging also showed that rAAV-luc/rAdv-luc cocktail vectors could synergistically enhance the early transient and latent sustained expression of luciferase, as compared to rAdv-luc and rAAV-luc alone. These findings suggest that the combination of rAAV-hTERTC27 and a therapeutic dose of rAdv-hTERTC27 is potentially a promising treatment for glioblastoma, and the rAAV/rAdv cocktail vector system warrants further development for cancer gene therapy.