Prospective phase II trial of nimotuzumab in combination with radiotherapy and concurrent capecitabine in locally advanced rectal cancer.

PURPOSE: The aim of the study was to evaluate the safety and efficacy of adding concurrent nimotuzumab to preoperative radiotherapy with concurrent capecitabine in locally advanced rectal cancer. METHODS AND MATERIALS: Patients with rectal cancer (clinical stage T3/4 or N+) were scheduled to receive weekly nimotuzumab (400 mg; days -6, 1, 8, 15, 22, and 29). Capecitabine (825 mg/m(2)) was delivered orally twice daily for the duration of radiotherapy. Radiotherapy was administered at 50.4 Gy (45 + 5.4 Gy). The main endpoint was the pathologic complete response (pCR) rate. RESULTS: Twenty-one patients with T3 or T4 disease were enrolled; 66.7 % were nodal-positive; the median distance from the anal verge was 5.5 cm. A pCR was achieved in four patients (19.0 %); 71.4 % patients obtained moderate or good tumor regression (Grade 2 and 3). Downstaging occurred in 15/21 (71.4 %) patients by T stage and 11/14 (78.6 %) by N stage. The actual dose intensities (median/mean, %) were nimotuzumab (100, 100) and capecitabine (100, 99.5). The most frequent Grade 1/2 toxicities were radiation dermatitis (57.1 %), nausea/vomiting (52.4 %), leukocytopenia (47.6 %), diarrhea (47.6 %), and proctitis (38.1 %). Grade 3 diarrhea was observed in 9.5 % of patients and Grade 3 leukocytopenia in 4.8 %. CONCLUSION: These preliminary results indicate that nimotuzumab can be safely combined with radiotherapy plus concurrent capecitabine. The efficacy of this regimen (pCR = 19.0 %) was significantly higher than that observed in previous phase II trials of preoperative radiotherapy with concurrent capecitabine and cetuximab in rectal cancer. Further investigation of concurrent nimotuzumab with radiotherapy plus capecitabine is warranted.

[Research progress in the study of protective effect of tanshinone IIA on cerebral ischemic stroke].

Danshen is one of the traditional Chinese herbal medicines and nas a long history or being used clinically in the treatment of cardiovascular and cerebrovascular conditions such as coronary heart disease and angina pectoris. Tanshinone IIA is a derivative of phenanthrene-quinone isolated from Danshen. It has been reported to be the major bioactive compound of Danshen and has diverse biological effects. Recent studies demonstrated that tanshinone IIA had neuroprotective effects on experimental ischemic stroke through its antiinflammatory, anti-oxidant, anti-apoptosis effects and its inhibitory effect on excitatory amino acid toxicity. In this review, we summarized all the recent progresses on the protective effect of tanshinone IIA on cerebral ischemic stroke. Hopefully, this article will throw some light on further study and application of tanshinone IIA.

[Effect of shenhuang ointment in promoting the rehabilitation of postoperative gastrointestinal dysfunction patients of Qi stagnation blood stasis syndrome: a clinical observation].

OBJECTIVE: To observe the effect and safety of plastering Chinese Compound Shenhuang Ointment (CSO) at Shenque (RN8) in promoting the rehabilitation of postoperative gastrointestinal dysfunction patients of qi stagnation blood stasis syndrome (QSBSS). METHODS: A prospective, multi-centered, randomized, double-blinded, controlled trial was conducted in 220 postoperative gastrointestinal dysfunction patients of QSBSS. They were randomly assigned to two groups, the CSO group (110 cases) and the placebo group (110 cases). CSO was plastered at Shenque (RN8) for 5 days after operation. The time of exhaustion, defecation, the recovery of intestinal peristalsis, integrals of TCM syndrome, and serum levels of motilin (MOT)and somatostatin (SS) were observed. RESULTS: Compared with the placebo group, the condition of exhaustion and defecation, the recovery of intestinal peristalsis on the 3rd day after operation was all improved (P < 0.05). The integrals of TCM syndrome at day 2, 3, and 4 were more significantly lowered in the CSO group than in the placebo group (P < 0.01, P < 0.05). The total effective rate of TCM syndrome was 95.3% in the CSO group, better than that in the placebo group (91.8%, P < 0.05). Compared with the placebo group, the serum MOT level increased and the serum SS level decreased at day 5 after operation in the CSO group (P < 0.05). CONCLUSIONS: The plastering of CSO at Shenque (RN8) could advance the time of exhaustion and defecation, and improve patients' clinical symptoms. And patients could tolerate well.

Primary Surgery Followed by Selective Chemoradiotherapy Versus Preoperative Chemoradiotherapy Followed by Surgery for Locally Advanced Rectal Cancer: A Randomized Clinical Trial.

PURPOSE: The aim of this work was to determine whether locally advanced rectal cancer (LARC) with negative mesorectal fascia (MRF) predicted by magnetic resonance imaging (MRI) can be excluded from preoperative radiation therapy treatment. METHODS AND MATERIALS: This multicenter, open-label, non-inferiority, randomized clinical trial enrolled patients with LARC within 6 to 12 cm from the anal verge and with negative MRI-predicted MRF. Participants were randomized to the intervention group (primary surgery, in which the patients with positive pathologic [CRM] circumferential margins were subjected to chemoradiotherapy [CRT] and those with negative CRM underwent adjuvant chemotherapy according to pathologic staging) or the control group (preoperative CRT, in which all patients underwent subsequent surgery and adjuvant chemotherapy). The primary endpoint was 3-year disease-free survival (DFS). RESULTS: A total of 275 patients were randomly assigned to the intervention (n = 140) and control (n = 135) groups, in which 33.57% and 28.15% patients were at clinical T4 stage and 85.92% and 80.45% patients were at "bad" or "ugly" risk in the intervention and control groups, respectively. There were 2 patients (1.52%) and 1 patient (0.77%) with positive CRM in the intervention and control groups, respectively (P > .05). The non-adherence rates for the intervention and control groups were 3.6% and 23.7%, respectively. After a median follow-up of 34.6 months (IQR, 18.2-45.7), 43 patients had positive events (28 patients and 15 patients in the intervention and control groups, respectively). There were 6 patients (4.4%) with local recurrence in the intervention group and none in the control group, which led to the termination of the trial. The 3-year DFS rate was 81.82% in the intervention group (95% CI, 78.18%-85.46%) and 85.37% in the control group (95% CI, 81.75%-88.99%), with a difference of -3.55% (95% CI, -3.71% to -3.39%; hazard ratio, 1.76; 95% CI, 0.94-3.30). In the per-protocol data set, the difference between 3-year DFS rates was -5.44% (95% CI, -5.63% to -5.25%; hazard ratio, 2.02; 95% CI, 1.01-4.06). CONCLUSIONS: Based on the outcomes of this trial, in patients with LARC and MRI-negative MRF, primary surgery could negatively influence their DFS rates. Therefore, primary surgery was an inferior strategy compared with preoperative CRT followed by surgery and cannot be recommended for patients with LARC.

[Double-mutated oncolytic adenovirus combined with gemcitabine for treating an orthotopic nude mouse model of bladder cancer].

OBJECTIVE: To investigate the therapeutic efficacy of double-mutated oncolytic adenovirus AxdAdB-3 in combination with gemcitabine for treating bladder cancer in an orthotopic nude mouse model. METHODS: The susceptibility to the adenovirus was evaluated in bladder cancer cell lines YTS-1, T24, 5637 and KK47, and normal cell lines HCV29 and WI38. The cells were infected with AxCAlacZ and stained with 5-bromo-4-chloro-3-indolyl-ß-galactoside (X-Gal). Immunostaining against adenoviral hexon protein was performed to determine the selective replication of AxdAdB-3 in the cancer cells. Flow cytometry was used to determine the YTS-1 cells in S phase of cell cycle after adenovirus infection. Cell viability after AxdAdB-3 and/or gemcitabine was measured by CCK-8 assay. Orthotopic bladder cancer model was established in nude mice, and the inhibitory efficacy of intravesical instillation therapy with AxdAdB-3 or/and gemcitabine was assessed. RESULTS: Gene transduction efficiency was different among the cell lines, and correlated with expression of CAR. 5637 and KK47 cells with high expression of CAR were more susceptible to the adenovirus, whereas YTS-1 and T24 cells with little CAR expression were resistant to adenoviral infection. Immunostaining showed that the expression levels of hexon protein varied among the cell lines. Normal cells infected with AxdAdB-3 expressed little hexon protein. The proportion of S-phase cells was (39 ± 3) % and (49 ± 5) % in the AxCAlacZ- and AxdAdB-3-infected bladder cancer cells, respectively. AxdAdB-3 effectively induced S-phase entry of cell cycle (P < 0.05). AxdAdB-3 combined with gemcitabine significantly inhibited the growth of bladder cancer cell lines. In vivo, the mean weight of the bladder tumors in mice treated with intravesical instillation of AxCAlacZ, gemcitabine, AxdAdB-3, and AxdAdB-3 + gemcitabine were 400.6, 126.4, 82. 0, 40.4 mg, respectively. Either AxdAdB-3 (P < 0.0001) and gemcitabine (P < 0.0001) suppressed the tumor growth in nude mice, and the combination therapy reduced tumors more effectively than either AxdAdB-3 (P < 0.0001) or gemcitabine (P < 0.0001) alone. CONCLUSIONS: Intravesical instillation therapy with AxdAdB-3 in combination with gemcitabine can effectively inhibit the orthotopic bladder cancer in nude mouse, and further relevant clinical studies are guaranteed.

[Detection and clinical significance of apoptosis related protein in local advanced rectal cancer patients with preoperative neoadjuvant therapy].

OBJECTIVE: To discuss the mechanism of rectal cancer apoptosis induced by preoperative chemoradiotherapy and evaluate its effect by detection of apoptosis related proteins in locally advanced colorectal cancer patients who had received preoperative chemoradiation. METHODS: To detect Bcl-XL and Bax expression in rectal cancer before and after chemoradiotherapy by EnVision method, combined with patients clinical and pathological index, statistically analysis and evaluation their relationship and clinical significance. RESULTS: Patients with or without tumor shrinkage after preoperative chemoradiotherapy was 13 cases and 21 cases. While the positive rate of Bcl-XL in rectal cancer before and after chemoradiotherapy were 58.8% (20/34) and 52.9% (18/34), respectively. There were significant difference between Bcl-XL change before and after chemoradiation with tumor size, tumor cells shrinkage and operation pattern. The positive rate of Bax in rectal cancer before and after chemoradiotherapy were 32.4% (11/34) and 44.1% (15/34), respectively. There were no significant difference between Bax change before and after chemoradiotherapy with tumor cells shrinkage. There were statistically significant difference between Bax ratio (χ(2) = 9.607, P = 0.048) before and after chemoradiation while there were no significant difference between Bcl-XL/Bax ratio before and after chemoradiation with tumor shrinkage. According to layered analysis with preoperative therapy, there were statistically significant difference (χ(2) = 13.964, P = 0.007) between Bcl-XL change with operation pattern while the same of significant difference between Bax change with tumor infiltration and tumor shrinkage (χ(2) = 10.806 and 10.455, both P < 0.05). CONCLUSIONS: Preoperative chemoradiation can influence rectal cancer cell's apoptosis and treatment effect by changing Bcl-XL and Bax expression. Bcl-XL downregulation and Bax upregulation have shown important function in colorectal cancer cell apoptosis which induced by preoperative chemoradiation, it can also improve the effection of chemoradiation in rectal cancer.

Computational Dissection of the Role of Trp305 in the Regulation of the Death-Associated Protein Kinase-Calmodulin Interaction.

Death-associated protein kinase 1 (DAPK1), as a calcium/calmodulin (CaM) regulated serine/threonine kinase, functions in apoptotic and autophagy pathways and represents an interesting drug target for inflammatory bowel disease and Alzheimer's disease. The crystal structure of the DAPK1 catalytic domain and the autoregulatory domain (ARD) in complex with CaM provides an understanding of CaM-dependent regulation of DAPK1 activity. However, the molecular basis of how distinct Trp305 (W305Y and W305D) mutations in the ARD modulate different DAPK1 activities remains unknown. Here, we performed multiple, µs-length molecular dynamics (MD) simulations of the DAPK1-CaM complex in three different (wild-type, W305Y, and W305D) states. MD simulations showed that the overall structural complex did not change significantly in the wild-type and W305Y systems, but underwent obvious conformational alteration in the W305D system. Dynamical cross-correlation and principal component analyses revealed that the W305D mutation enhanced the anti-correlated motions between the DAPK1 and CaM and sampled a broader distribution of conformational space relative to the wild-type and W305Y systems. Structural and energetical analyses further exhibited that CaM binding was unfavored in response to the W305D mutation, resulting in the decreased binding of CaM to the W305D mutant. Furthermore, the hydrogen bonds and salt bridges responsible for the loss of CaM binding on the interface of the DAPK1-CaM complex were identified in the W305D mutant. This result may provide insights into the key role of Trp305 in the regulation of CaM-mediated DAPK1 activity.

[Efficacy and toxicity analysis of XELOX and FOLFOX4 regimens as adjuvant chemotherapy for stage III colorectal cancer].

OBJECTIVE: To compare the efficacy and toxicity of capecitabine plus oxaliplatin (XELOX) versus 5-fluorouracil/leucovorin (5-Fu/LV) plus oxaliplatin (FOLFOX4) regimens as adjuvant chemotherapy for stage III colorectal cancer. METHODS: The clinicopathological data of 118 patients with stage III colorectal cancer were studied retrospectively. The patients were assigned to receive either FOLFOX4 regimen (n = 76) or XELOX regimen (n = 42). 3-year disease-free survival (DFS) and adverse events as end points were compared between the two groups. RESULTS: The number of patients that failed to finish 8 cycles was higher in FOLFOX4 group (28 vs. 8, P = 0.044). There was no significant difference for 3-year DFS and all grades adverse events between the two groups. However, the FOLFOX4 group showed more grade 3/4 neutropenia (31.6% vs. 14.3%, P = 0.039) and central venous catheter-associated complication (11.8% vs. 4.8%, P = 0.205), while XELOX showed more grade 3/4 thrombocytopenia (19.0% vs. 6.6%, P = 0.038) and hand-foot syndrome (11.9% vs. 1.3%, P = 0.012). CONCLUSION: The results of this analysis indicate that XELOX and FOLFOX4 regimens have very similar efficacy as an adjuvant chemotherapy for stage III colon cancer, but XELOX may be safer than FOLFOX4.

Circulating guanylyl cyclase C (GCC) mRNA is a reliable metastatic predictor and prognostic index of colorectal cancer.

BACKGROUND: Currently, few specific biomarkers or standard cutoff values are available for circulating tumor cells (CTCs) detection and survival prediction in patients with early stage colorectal cancer (CRC). Guanylyl cyclase C (GCC) presents as a specific expression in intestinal tumor cells and during their metastases, indicating its potential application as a metastatic predictor of CRC. METHODS: The circulating GCC mRNA of 160 colorectal cancer patients at stage I-III was detected via quantitative real-time (qRT)-PCR in our study, and the correlation of GCC mRNA level with tumor metastasis and long-term survival was explored. RESULTS: GCC mRNA was found to be positive in 43 out of 160 CRC patients and negative in ten healthy controls. It was found that GCC mRNA over the baseline (>100 copies/µL and 200 copies/µL) showed a significant correlation with disease-free survival (DFS) and overall survival (OS) in the stage II subgroup. It was further revealed that GCC mRNA over 300 copies/µL or higher than the median value of copy numbers was significantly correlated with reduced OS and DFS in CRC patients. A nomogram model based on variables including GCC mRNA copy number was established for predicting the OS of CRC patients (AUC =0.98). CONCLUSIONS: Circulating GCC mRNA over baseline is a reliable predictor for tumor metastasis and can be a prognostic index in CRC patients.

[Prognostic significance of metastatic lymph node ratio in colorectal cancer].

OBJECTIVE: To investigate the prognostic significance of metastatic lymph node ratio in patients with colorectal cancer. METHODS: The clinicopathological data of 303 surgically treated patients with colorectal cancer were retrospectively analyzed. Spearman correlation analysis was used to determine the correlation coefficient. The survival was analyzed using Kaplan-Meier method, and the survival difference was assessed by Log-rank test. Multivariate analysis was performed using Cox proportional hazard regression model in forward stepwise regression. Receiver working characteristic curve was used to compare the accuracy of the metastatic lymph nodes ratio in predicting the death of patients at 5 years postoperatively with that of the number of metastatic lymph nodes. RESULTS: The MLR was not correlated with the total number of dissected lymph nodes (Spearman correlation coefficient: -0.099, P > 0.05), but the positive rate of metastatic lymph nodes did (correlation coefficient: 0.107, P < 0.05). Kaplan-Meier survival analysis revealed that the MLR significantly influenced the postoperative survival time (Log-rank chi(2) = 42.878, P < 0.01), even in the patients with less than 12 resected lymph nodes. The 5-year survival rates for rN0, rN1, rN2 and rN3 were 90.9%, 68.9%, 54.7% and 39.4%, respectively. There was a significant difference between the different stages (P < 0.01). Cox proportional hazard regression model analysis showed that the metastatic lymph node ratio was an independent prognostic factor. (EXP(B) = 7.809, P < 0.01). There was no significant difference between metastatic lymph node ratio and the number of metastatic lymph nodes in predicting the death of patients at 5 years postoperatively based on the area under the receiver working characteristic curve. CONCLUSION: The metastatic lymph node ratio in colorectal cancer patients is not correlated with the total number of dissected lymph nodes. The metastatic lymph node ratio is a major independent prognostic factor for patients with colorectal cancer. The ability of metastatic lymph node ratio in predicting the death of colorectal cancer patients at 5 years postoperatively is the same as that of the number of metastatic lymph nodes.

Conversion chemotherapy with capecitabine and oxaliplatin for colorectal cancer with potentially resectable liver metastases: A phase II, open-label, single-arm study.

AIM: The aim of this is study is to assess the efficacy and safety of conversion capecitabine plus oxaliplatin (XELOX) in Chinese patients with potentially resectable colorectal liver metastases (CLMs). PATIENTS AND METHODS: Thirty patients (median age 57.5 years) with potentially resectable CLMs were treated with XELOX in a single-arm, open-label, nonrandomized, multicenter clinical trial. RESULTS: The objective response rate in the 30 patients was 40% (95% confidence interval: 22.7%-59.4%), and the rate of conversion to resectable CLMs was 43.3%. Patients who underwent liver resection (n = 11) had a longer median progression-free survival and overall survival than those who did not. XELOX showed an acceptable safety profile. CONCLUSION: XELOX may effectively convert potentially resectable CLM into resectable CLM, providing survival benefits with a favorable safety profile. CLINICAL TRIALS.GOV IDENTIFIER: NCT 00997685.

[Effects of enterostomy in treating locally advanced rectal cancer with combined chemoradiotherapy and operation].

OBJECTIVE: To investigate the effect of enterostomy in treatment of locally advanced rectal carcinoma patients with combined chemoradiotherapy and operation. METHODS: Clinical data from 51 cases of locally advanced rectal cancer patients treated with preoperative chemoradiotherapy and operation were analyzed. RESULTS: Thirty-three patients (64.9%) got staging down of their cancer after preoperative chemoradiotherapy, and 21.6% of patients (11 cases) had complete pathologic response. Thirty-seven patients received enterostomy, including extraperitoneal sigmoidostomy (29 cases), defunctioning ileostomy (8 cases) and double colostomy (3 cases with colon obstruction during preoperative therapy). One case experienced parastomal hernia and one stomal stenosis and 2 cases parastomal infection after enterostomy. No death of enterostomy occurred. CONCLUSION: Colostomy can reduce the pressure of obstructed intestinal tract and contribute much to the preoperative chemoradiotherapy, ileostomy can protect the distal stoma from leakage in sphincter saving operation. Enterostomy could be selected when needed in the favor of locally advanced rectal cancer patients.

Smad3 mutant mice develop colon cancer with overexpression of COX-2.

Colon cancer is the second most common cause of cancer-associated mortality in human populations. The aim of the present study was to identify the role of cyclooxygenase-2 (COX-2) in Smad3 mutant mice, which are known to develop colon cancer. Homozygous Smad3 (-/-) mutant mice were generated from inbred and hybrid Smad3 mouse strains by intercrossing the appropriate heterozygotes. Immunohistochemistry with COX-2 antibody was performed throughout this experiment and the data was validated and cross-checked with reverse transcription-polymerase chain reaction (RT-PCR). Homozygous mutant Smad3 mice were generated and the overexpression pattern of COX-2 was identified by immunohistochemistry and validated with RT-PCR. The results of the present study demonstrated a link between the Smad3 mutant mice, colon cancer and COX-2. In addition, the overexpression pattern of COX-2 in Smad3 mutant mice that develop colon cancer was identified.

A novel synthetic derivative of squamosamide FLZ inhibits the high mobility group box 1 protein-mediated neuroinflammatory responses in murine BV2 microglial cells.

High mobility group box 1 (HMGB1) is a critical pro-inflammatory cytokine that contributes to the pathogenesis of various human diseases. FLZ, a squamosamide derivative, has been demonstrated to have neuroprotective effects in Parkinson's disease models and shows strong anti-inflammatory activity, while the precise mechanism remains unclear. Here, we investigated the anti-inflammatory mechanism of FLZ on HMGB1-mediated inflammatory responses. The effects of FLZ on HMGB1 release from microglial cells induced by lipopolysaccharide were first explored by Western blot assay and ELISA. Then, co-immunoprecipition was used to study FLZ's effect on the interaction between HMGB1 and its receptor TLR4. Finally, we employed HMGB1 to simulate pro-inflammatory responses and then studied the inhibitory effects of FLZ on its bioactivity. FLZ has a significant inhibitory effect on HMGB1 release while it exerts no inhibitory effect on the binding between HMGB1 and TLR4. After the recognition of HMGB1 by TLR4, NF-κB signaling pathway is activated. FLZ could efficaciously alleviate HMGB1-induced inflammatory responses via the suppression of TLR4/MyD88/NF-κB signaling pathway. FLZ could inhibit HMGB1 release as well as HMGB1-induced inflammatory responses, HMGB1 might be one of the FLZ anti-inflammatory targets, and interfering at this inflammatory mediator may have benefit effects on neurodegenerative disorders, such as Parkinson's disease.

Interleukin-11-induced capillary leak syndrome companied with abdominal chylous leakage in primary sigmoid carcinoma patients with thrombocytopenia.

Capillary leak syndrome (CLS) is a rare condition characterized by generalized edema and hypotension. Interleukin-11 (IL-11) is a therapeutic agent for the treatment of thrombocytopenia. The relationship between IL-11 and CLS has rarely been reported, especially in patients with colorectal cancer. We describe a case with sigmoid cancer treated with IL-11 after chemotherapy. After 5 days of IL-11 therapy, the patient felt tachypnea, muscular pain and fullness of the abdomen. Chest X-ray indicated increased bronchovascular shadows, and abdominal ultrasound indicated moderate ascites. IL-11 was then discontinued, fluid resuscitation was performed, and fresh frozen plasma and packed red blood cells were transfused. On the fourth day, synchronous chylous leakage was detected. Low fat diet, nutritional support, and somatostatin was administered. The patient recovered 2 weeks later. Although rare, CLS could be a severe side effect after the administration of IL-11. The aim of treatment is to stabilize the vital parameters.

[Detection of guanylate cyclase C mRNA and cytokeratin 20 mRNA in peripheral blood and analysis of prognosis in early to moderate colorectal cancer patients].

OBJECTIVE: To investigate the associations of guanylate cyclase C (GC-C) mRNA and cytokeratin 20 (CK20) mRNA with metastasis and prognosis in early to moderate colorectal cancer patients. METHODS: GC-C mRNA and CK 20 mRNA in peripheral blood of 74 colorectal cancer patients without distant metastasis were detected by fluorescent quantitative PCR (FQ-PCR). Based on their clinicopathological and postoperative follow-up data, the relationship and clinical significance of these data with metastasis hazards and prognosis factors were analyzed. RESULTS: The positive rate of GC-C mRNA in 74 colorectal cancer patients was 33.8% (25/74), and CK20 mRNA was 31.1% (23/74). The 1-, 2-, 3- year disease-free survival rates of patients were 94.6%, 82.4% and 78.4% respectively. There were significant differences in positive rates of GC-C mRNA and CK20 mRNA, tumor differentiation, mesentery lymph node metastasis, tumor embolus in vessel and postoperative chemotherapy associated with 3-year disease free survival rate by Kaplan-Meier analysis (all P<0.05). While mesentery lymph node metastasis and tumor embolus in vessel were independent risk factors of 3-year disease-free survival (P<0.05). CK20 mRNA and tumor embolus in vessel were independent risk factors of 3-year disease-free survival by analysis stratified with clinical stage (P<0.05). CONCLUSIONS: Detection of CK20 mRNA and GC-C mRNA in peripheral blood may be important for early detection of early metastasis of colorectal cancer.

Detection of circulating tumor cells in peripheral blood of colorectal cancer patients without distant organ metastases.

PURPOSE: Recently, the detection of circulating tumor cells (CTCs) in peripheral blood has become an important tool for the non-invasive assessment of micrometastases and to predict clinical outcome. The objective of this study was to investigate if the presence of CTCs in peripheral blood influences the prognosis in colorectal cancer (CRC) patients without distant organ metastases. METHODS: The GCC mRNA and CK20 mRNA levels in peripheral blood and the serum levels of CEA of 92 CRC patients without distant organ metastasis were analyzed by quantitative RT-PCR and ELISA, respectively. Its associations with overall survival (OS) and disease-free survival (DFS) rates were analyzed. RESULTS: Univariate analyses showed that lower OS and DFS rates were significantly associated with GCC and CK20 mRNA levels, the presence of lymph node metastases, the presence of mesenteric root lymph node metastases, and the presence of tumor emboli in vessels (p < 0.05), but not with CEA levels. Multivariate analyses showed a significant association between 1) OS and GCC mRNA levels and differentiation types and 2) DFS and the presence of tumor emboli in the vessels. Kaplan-Meier curves showed that DFS was significantly associated with the presence of poorly differentiated cells, the presence of mesenteric root lymph node metastases having received prior chemotherapy, and the presence of tumor emboli in vessels. CONCLUSION: The detection of CTCs in peripheral blood may be useful for the prediction of clinical outcome in CRC patients without distant organ metastases.

[Anti-tumor effect of adenovirus-mediated Bcl-XL shRNA in vitro].

OBJECTIVE: To investigate the anti-tumor effect of adenovirus-mediated Bcl-XL shRNA on colon cancer cells in vitro. METHODS: A recombinant Bcl-xl adenovirus was constructed, amplified, and purified. The effect on mRNA expression of Bcl-XL was assessed by RT-PCR, and the effect on apoptosis-induction of colon cancer(Lovo cell line) in vitro was assessed by MTT assay and cell clonogenic assay. RESULTS: RT-PCR showed that Ad/Bcl-XL shRNA significantly down-regulated the mRNA expression of Bcl-XL in Lovo cells. Ad/Bcl-XL shRNA suppressed the proliferation of Lovo cells in a dose-dependent as well as a time-dependent manner compared with Ad/GFP (P<0.05). Treatment with Ad/Bcl-XL shRNA dramatically suppressed the colony formation of Lovo cells in a dose-dependent manner (P<0.05). Ad/Bcl-XL shRNA showed no effect on normal human fibroblast. CONCLUSION: Ad/Bcl-XL shRNA exhibits cytotoxic effect on Lovo cells and may have the potential value in the treatment of colon cancer.

E1A, E1B double-restricted adenovirus enhances the cytotoxicity and antitumor activity of gemcitabine to renal cell carcinoma.

BACKGROUND: Our previous studies have demonstrated potent oncolysis efficacy of the E1A, E1B double-restricted replication-competent oncolytic adenovirus AxdAdB-3 for treatment of bladder cancer. Here, we reported the feasibility and efficacy of AxdAdB-3 alone, or in combination with gemcitabine for treating renal cell carcinoma. METHODS: Cytopathic effects of AxdAdB-3 were evaluated in human renal cell carcinoma cell lines TOS-1, TOS-2, TOS-3, TOS-3LN, SMKT-R3, SMKT-R4 and ACHN, and in normal human renal proximal tubule epithelial cells (RPTEC). AxdAdB-3 induced down-regulation of the cell cycle was determined by flow cytometry. Combination therapies of AxdAdB-3 with gemcitabine were evaluated in vitro and in vivo on subcutaneous TOS-3LN tumors in a severe combined immunodeficiency disease (SCID) mouse model. RESULTS: AxdAdB-3 was potently cytopathic against the tested most renal cell carcinoma cell lines including TOS-2, TOS-3, TOS-3LN, SMKT-R3 and SMKT-R4, while normal human RPTEC were not destroyed. AxdAdB-3 effectively induced cell cycle S-phase entry. Combined therapy of AxdAdB-3 with gemcitabine demonstrated stronger antitumor effects in vitro and in vivo compared with either AxdAdB-3 or gemcitabine alone. CONCLUSION: AxdAdB-3 alone, or in combination with gemcitabine may be a promising strategy against renal cell carcinoma.

[Differential expression of guanylin in colorectal cancer].

OBJECTIVE: To investigate the expression of guanylin in colorectal cancer. METHODS: The expression of guanylin was examined by RT-PCR and semiquantitative analysis in 20 cases of colorectal cancer, and its relationship with clinical characteristics was analyzed. RESULTS: The positive expression of guanylin in normal tissue (80%, 16/20) was significantly higher than that in tumor tissue (35%, 7/20) (P<0.01). The same result was found in the semiquantitative analysis of 14 cases with differential expression. Differential expression of guanylin in colorectal cancer was associate with TNM stage (P<0.05), not with sex, Borrmann type and degree of differentiation (all P>0.05). CONCLUSION: There is differential expression of guanylin in colorectal cancer, and this kind of differential expression is associated with tumor TNM stage.