Potent bromodomain and extraterminal domain inhibitor JAB-8263 suppresses MYC expression and exerts anti-tumor activity in colorectal cancer models.

BACKGROUND: The overexpression of the MYC gene plays an important role in the occurrence, development and evolution of colorectal cancer (CRC). Bromodomain and extraterminal domain (BET) inhibitors can decrease the function BET by recognizing acetylated lysine residues, thereby downregulating the expression of MYC. AIM: To investigate the inhibitory effect and mechanism of a BET inhibitor on CRC cells. METHODS: The effect of the BET inhibitor JAB-8263 on the proliferation of various CRC cell lines was studied by CellTiter-Glo method and colony formation assay. The effect of JAB-8263 on the cell cycle and apoptosis of CRC cells was studied by propidium iodide staining and Annexin V/propidium iodide flow assay, respectively. The effect of JAB-8263 on the expression of c-MYC, p21 and p16 in CRC cells was detected by western blotting assay. The anti-tumor effect of JAB-8263 on CRC cells in vivo and evaluation of the safety of the compound was predicted by constructing a CRC cell animal tumor model. RESULTS: JAB-8263 dose-dependently suppressed CRC cell proliferation and colony formation in vitro. The MYC signaling pathway was dose-dependently inhibited by JAB-8263 in human CRC cell lines. JAB-8263 dose-dependently induced cell cycle arrest and apoptosis in the MC38 cell line. SW837 xenograft model was treated with JAB-8263 (0.3 mg/kg for 29 d), and the average tumor volume was significantly decreased compared to the vehicle control group (P < 0.001). The MC38 syngeneic murine model was treated with JAB-8263 (0.2 mg/kg for 29 d), and the average tumor volume was significantly decreased compared to the vehicle control group (P = 0.003). CONCLUSION: BET could be a potential effective drug target for suppressing CRC growth, and the BET inhibitor JAB-8263 can effectively suppress c-MYC expression and exert anti-tumor activity in CRC models.

miR-887-3p Inhibits the Progression of Colorectal Cancer via Downregulating DNMT1 Expression and Regulating P53 Expression.

Colorectal cancer (CRC) is the third most diagnosed cancer worldwide and the second leading cause of cancer-related deaths. Many researchers have reported that abnormal microRNAs (miRs) were expressed in CRC and participated in the occurrence and progression of CRC. However, there are few reports of miR-887-3p regulating CRC development. In the current study, we investigated the abnormal expression of miR-887-3p and also demonstrated its regulatory role and detailed molecular mechanism in CRC. Initially, miRNA expression data were obtained from TCGA-COAD that consisted of 453 CRC samples and 8 normal tissue samples. These were downloaded and analyzed to compare the expression level of miR-887-3p in CRC tissues to that in normal tissues. Moreover, 32 pairs of surgically resected CRC tumors and para-cancer tissues from our hospital were collected. Quantitative real-time PCR (qRT-PCR) was performed to detect miR-887-3p expression levels in CRC tissues, para-cancer tissues, several CRC cell lines, and an intestinal epithelial cell line. Following miR-887-3p mimic transfection in colon cancer SW480 cell line, the regulatory roles of miR-887-3p on cell proliferation, apoptosis, invasion, migration, and epithelial-mesenchymal transition (EMT) were detected through CCK-8, flow cytometry, transwell assay, and Western blot. After potential targeting protein was predicted by bioinformatic websites, the luciferase reporter assay and Western blot were used to confirm the target of miR-887-3p. The targeting protein expressions were detected by Western blot and qRT-PCR. The relationship between miR-887-3p level and the effect of miR-887-3p on P53 expression was evaluated by Western blot and qRT-PCR. The effects of miR-887-3p on CRC cell growth in vivo by xenograft tumor experiments were investigated, and Ki-67 in tumor tissue was determined by immunohistochemistry. Results. The COAD data demonstrated that the expression levels of miR-887-3p in CRC clinical sample tissues and cell line cultures were remarkably lower than para-cancer normal tissues and NCM460 cells (normal colonic epithelial cell line). Functional experiments demonstrated that overexpression of miR-887-3p in SW480 cells significantly reduced proliferation, migration, invasion, and EMT, and promoted cancer cell apoptosis. Additionally, Western blot, qRT-PCR, and luciferase reporter assays confirmed that DNMT1 was a downstream target of miR-887-3p. Moreover, the blocking of DNMT1 by miR-887-3p mimics also promoted P53 expression. Finally, overexpression of DNMT1 in SW480 cells could partially reverse the regulatory effect of miR-887-3p mimics on CRC cell development. From in vivo experiments, overexpression of miR-887-3p could inhibit tumor growth in CRC xenograft mice and reduce the Ki-67 level. Conclusion. The microRNA miR-887-3p is a potential biomarker of CRC. It inhibited CRC cell proliferation, invasion, and EMT, and promoted cell apoptosis through targeting and downregulating DNMT1 and promoting P53 expression. Therefore, miR-887-3p may be a good biomarker and therapeutic target for CRC treatment.

Circular RNA circ_0006089 regulates the IGF1R expression by targeting miR-143-3p to promote gastric cancer proliferation, migration and invasion.

Circular RNAs (circRNAs) figure prominently in regulating the progression of a variety of human malignancies. This study was performed to probe how circ_0006089 functioned in gastric cancer (GC). CircRNA expression profile GSE83521 was downloaded from Gene Expression Omnibus (GEO) database, and circRNAs and analyzed. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure circ_0006089, microRNA-143-3p (miR-143-3p) and insulin-like growth factor 1 receptor (IGF1R) mRNA expressions in GC tissues and cell lines. Kaplan-Meier curves were used to detect the relationship between circ_0006089 expression and overall survival time of GC patients. Cell counting kit-8 (CCK-8) and 5-bromo-2-deoxyuridine (BrdU) assays were employed to detect the proliferative ability of GC cells after circ_0006089 was overexpressed or knocked down. Wound healing assay and Transwell assay were executed to examine the migration and invasion ability of GC cells. Western blot was adopted to detect IGF1R protein expressions. Circ_0006089 expression was up-regulated in GC samples and cell lines. And high circ_0006089 expression was associated with shorter survival time in GC patients. Circ_0006089 overexpression in GC cells significantly accelerated GC cell proliferation, migration and invasion, whereas circ_0006089 knockdown resulted in the opposite effects. Additionally, miR-143-3p was validated as a downstream target of circ_0006089, and circ_0006089 could positively regulate IGF1R expression via repressing miR-143-3p. Circ_0006089 is highly expressed in GC, and it promotes the malignancy of GC cells via modulating miR-143-3p/IGF1R axis, suggesting that circ_0006089 may serve as a promising therapeutic target for GC.

Analysis of Threshold Changes of Tumor Mutation Burden of Gastric Cancer and Its Relationship with Patients' Prognosis.

OBJECTIVE: Gastric cancer is a malignant tumor originating from gastric mucosal epithelium. Here, we aimed to investigate the analysis of the threshold change of gastric cancer tumor mutation burden (TMB) and its relationship with the prognosis of patients. METHODS: 256 patients with gastric cancer were selected as subjects. All patients were in the advanced stage and received surgical resection of D2 lymph node dissection. After the operation, a follow-up was performed for 24 months, and the disease-free survival and overall survival of patients were counted. The NGS molecular biological was detected to obtain gastric cancer tumor mutation burden (TMB) data. Pearson correlation analysis software was used to analyze the correlation between TMB threshold and disease-free survival or overall survival of patients with gastric cancer, and the multivariate logistic analysis was performed as well. RESULTS: The disease-free survival period and the overall survival period of patients in the low-to-medium TMB group were both longer than those in the high TMB group. Pearson correlation analysis results showed that the TMB threshold was negatively correlated with the disease-free survival and overall survival of gastric cancer patients. Results from multivariate logistic analysis showed that high TMB thresholds have a greater impact on disease-free survival and overall survival of patients, but the impact of medium and low TMB thresholds on disease-free survival and overall survival of patients is weakened. CONCLUSIONS: The TMB threshold level has a predictive effect on the effect of surgical resection of D2 lymph node dissection, and high levels of TMB can significantly affect disease-free survival and overall survival of patients with advanced gastric cancer.

Resection outcomes for primary and local recurrent retroperitoneal liposarcoma patients.

BACKGROUND: The clinical characteristics of primary retroperitoneal liposarcoma (PR RPLPS) and local recurrent retroperitoneal liposarcoma (LR RPLPS) cases were compared to determine the related factors involved in postoperative survival. METHODS: A total of 90 patients who underwent surgery between 2006 and 2013 were included in this study. Clinicopathological data that was prospectively gathered was analyzed to identify factors associated with overall survival (OS) and progression-free survival (PFS). RESULTS: The PR cases showed a higher complete resection rate when compared to the LR group. The LR group showed a greater number that were poorly differentiated and highly malignant. More blood loss was observed in the LR compared to the PR group. Multivariate analysis suggested that blood loss and tumor grade were prognostic factors for OS and PFS of the PR group, but extent of resection was a prognostic factor only for OS. In the LR group, the extent of resection was a significant prognostic factor associated with OS, whereas tumor grade was associated with PFS. CONCLUSIONS: Complete surgical resection is the most important factor for the survival of RPLPS patients. Tumor grade is an independent prognostic factor for PFS. In PR RPLPS, poor tumor classification and increased intraoperative bleeding are associated with a poor prognosis.

The impact of COVID-19 on gastric cancer surgery: a single-center retrospective study.

BACKGROUND: The coronavirus disease 2019 (COVID-19) has been declared a global pandemic by the World Health Organization. Patients with cancer are more likely to incur poor clinical outcomes. Due to the prevailing pandemic, we propose some surgical strategies for gastric cancer patients. METHODS: The 'COVID-19' period was defined as occurring between 2020 and 01-20 and 2020-03-20. The enrolled patients were divided into two groups, pre-COVID-19 group (PCG) and COVID-19 group (CG). A total of 109 patients with gastric cancer were enrolled in this study. RESULTS: The waiting time before admission increased by 4 days in the CG (PCG: 4.5 [IQR: 2, 7.8] vs. CG: 8.0 [IQR: 2,20]; p = 0.006). More patients had performed chest CT scans besides abdominal CT before admission during the COVID-19 period (PCG: 22 [32%] vs. CG: 30 [73%], p = 0.001). After admission during the COVID period, the waiting time before surgery was longer (PCG: 3[IQR: 2,5] vs. CG: 7[IQR: 5,9]; p < 0.001), more laparoscopic surgeries were performed (PCG: 51[75%] vs. CG: 38[92%], p = 0.021), and hospital stay period after surgery was longer (7[IQR: 6,8] vs.9[IQR:7,11]; p < 0.001). In addition, the total cost of hospitalization increased during this period, (PCG: 9.22[IQR:7.82,10.97] vs. CG: 10.42[IQR:8.99,12.57]; p = 0.006). CONCLUSION: This study provides an opportunity for our surgical colleagues to reflect on their own services and any contingency plans they may have to tackle the COVID-19 crisis.

Continuous Hemofiltration Reduces Mortality in Severe Acute Pancreatitis: A Meta-Analysis.

BACKGROUND: Severe acute pancreatitis (SAP) is a deadly condition, with a mortality rate ranging from 15% to 30%. Recently, blood purification therapy has been adopted in administrating SAP patients. The present study aimed at evaluating the effect of continuous hemofiltration therapy for SAP. METHODS: A systematic search of Cochrane Library, PubMed, and Embase was carried out until October 1st, 2019. Prospective studies comparing outcomes for SAP patients between continuous hemofiltration and standard therapy were enrolled. RESULTS: Continuous hemofiltration therapy was associated with lower level of PACHE II score (MD = -1.49; 95% CI: -2.69 to -0.29, P=0.02), CRP (MD = -1.56 mg/L; 95% CI: -2.64 to -0.47, P=0.005), Cr (MD = -3.57 umol/L; 95% CI: -5.50 to -1.65, P=0.003), and Bun (MD = -3.63 mmol/L; 95% CI: -6.07 to -1.20, P=0.003) at 72 h after onset of treatment. Continuous hemofiltration therapy was associated with shorter length of abdominal pain relief time (MD = -1.82 hours; 95% CI: -2.93 to -0.71, P=0.001), lower surgery rate (OR = 0.15; 95% CI: 0.03 to 0.78, P=0.02), and mortality rate (OR = 0.54; 95% CI: 0.37 to 0.77, P=0.0007). CONCLUSIONS: continuous hemofiltration therapy could effectively alleviate SAP as early as 72 hours after onset of treatment, lowering the level of Bun, Cr, CRP, and APACHE II scores. Continuous hemofiltration therapy could confer SAP patients with lower mortality rates.

Serum levels of PTEN mRNA in colorectal cancer: a case-control study.

Background: The purpose of this study was to identify the expression of phosphatase and tensin homolog (PTEN) and its diagnostic value in colorectal cancer (CRC).Methods: The expression level of serum PTEN at mRNA and protein level were determined using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analyses, respectively. Chi-square test was employed to explore the relationship between PTEN expression and clinical features of CRC patients. The receiver operating characteristics (ROC) curve was established to evaluate the diagnostic performance of PTEN in CRC.Results: The expression of serum PTEN was significantly lower in patients with CRC than that in healthy controls both at mRNA and protein level (p < .05). Also, the low PTEN expression was significantly related to serosal invasion, lymph node metastasis and Ki-67, but had no relation with age, sex, tumor depth and tumor site. The area under ROC curve of 0.810 corresponding with a sensitivity of 97.79% and a specificity of 70.31% were obtained, which suggested PTEN could act as a diagnostic marker for CRC.Conclusion: Altogether, serum PTEN expression was down-regulated and it participated in the development of CRC. Besides, it could act as an efficient and independent diagnostic marker for CRC patients.

Treatment Effect of Tuftsin and Antigen Peptide Combined with Immune Cells on Colorectal Cancer.

BACKGROUND The aim of this study was to investigate the effect of antigenic peptides on dendritic cell maturation and activation as well as the role of dendritic cell induced cell function. The tumor-specific cytotoxic T lymphocytes induced by activation of the dendritic cells were also evaluated. MATERIAL AND METHODS SW-480 cell lysate and peptide antigens were selected as adjuvants in dendritic cell sensitization, and tuftsin was used to induce the phagocytosis of dendritic cells. Immature dendritic cells were stimulated with the antigen and adjuvant as follows: group A was negative control; group B was SW-480 (20 µg/mL); group C was SW-480 (20 µg/mL)+tumor necrosis factor (TNF)-alpha (10 µg/mL); group D was SW-480 (20 µg/mL)+tuftsin (20 µg/mL); group E was antigen peptide (2 µg/mL); group F was antigen peptide (2 µg/mL)+TNF-alpha (10 µg/mL); group G was antigen peptide (2 µg/mL)+tuftsin (20 µg/mL). Cytotoxic T lymphocytes activation and in vitro anti-tumor effects were examined by detecting the maturation marks of dendritic cells as well as interleukin (IL)-10 and IL-12 levels secreted by dendritic cells. Cells with the strongest immunizing effects were injected into nude mice and tumor suppression status was evaluated. RESULTS Group D (SW-480+tuftsin), group E (antigen peptides), group F (antigen peptide+TNF-alpha), and group G (antigen peptides+tuftsin) displayed significant differences compared to the control group (P<0.05). Group G (antigen peptides+tuftsin) could also promote the secretion of cytokines IL-12, as well as inhibit cytokine IL-10 secretion, compared to the other experimental groups (P<0.05). In the in vivo experiments of tumor inhibitions, antigenic polypeptide+tuftsin was the most effective (P<0.05). CONCLUSIONS Combination of cytotoxic T lymphocytes and T peptide therapy in treating human colorectal cancer might be used as a new treatment strategy based on adoptive cellular immunotherapy.

Effect of TGF-ß1 on Apoptosis of Colon Cancer Cells Via the ERK Signaling Pathway.

PURPOSE: To study the effect of transforming growth factor (TGF)-ß1 on apoptosis of colon cancer cells via the ERK signaling pathway. METHODS: Human chemosensitive colon cancer cell line HT- 29 was used in this study. VEGF mRNA and protein level were detected using PCR and western blot. Enzyme-linked immunosorbent assay (ELISA) was used for prostaglandin (PG) detection. Cell proliferation was determined via MTT assay. RESULTS: TGF-ß1 had a significant effect on blocking the cancer cell growth (p<0.05). TGF-ß1 significantly reduced the VEGF mRNA level (p<0.05) and eliminated the COX-2 expression in a dose-dependent manner, while p53 expression was increased (p<0.05). TGF-ß1-induced inhibitory effect on COX-2 was significantly eliminated by the ERK inhibitor Compound C (p<0.05). The basal PGE2 production was eliminated in cells treated with TGF-ß1 (p<0.05). N-acetylcysteine (NAC) treatment almost completely eliminated the reactive oxygen species (ROS) produced by TGF-ß1 and ERK activation. Compared with administration of 5-FU or etoposide alone, TGF-Β1 combined with 5-FU or etoposide significantly administration the viability of colon cancer HT-29 cells. CONCLUSION: ERK is a newly-identified cancer target molecule, which can significantly control COX-2 in colon cancer cells treated with TGF-ß1.

Application of highly efficient and lowly toxic bufadienolides screened from toad skin in lymphatic chemotherapy for colorectal cancer through a lymphatic metastatic model.

BACKGROUND: Lymph node metastasis (LNM) remains a major obstacle to treat colorectal cancer (CRC). Increasing evidences have suggested that bufadienolides contain several fractions displaying antitumor activity and may be applied in lymphatic chemotherapy. However, effects of the highly efficient and lowly toxic (HELT) bufadienolides on CRC in lymphatic chemotherapy have not been reported. METHODS: Adenosine triphosphate tumor chemosensitivity assays (ATP-TCA) was performed to detect the inhibition rate (IR) of fractions of bufadienolides to cytokine-induced killer (CIK) cells and tumor cells. HELT fraction-loaded emulsions of different concentrations were prepared. Nude mouse bearing HCT116 tumors in footpad received high-dose emulsion (HD-E), middle-dose emulsion (MD-E), low-dose emulsion (LD-E), control emulsion (CE), Cinobufacini Injection (CI), or normal saline (NS), respectively. Hematoxylin and eosin (H&E) staining, Flow Cytometry (FCM), enzyme-linked immune sorbent assay (ELISA) and hematological examination were applied to evaluate therapeutic effects and potential toxicity. RESULTS: F18 and F19 were screened out as HELT fractions in vivo and F18-loaded emulsions of different concentrations for lymphatic administration were prepared. We confirmed that HD-E and MD-E produced obvious antitumor activities in footpad tumors and LNM compared with other groups in vitro. We also verified the effects of F18-loaded emulsions on activating hematopoietic function, stimulating proliferation of the spleen and natural killer (NK) cells, and promoting the secretion of IFN-γ and IgG1, although HD-E performed mild toxicity on liver. CONCLUSION: The present study demonstrated that lymphatic chemotherapy with HELT fraction of bufadienolides could be an effective approach to the treatment of CRC patients with LNM.

MicroRNA-150 inhibits the proliferation and metastasis potential of colorectal cancer cells by targeting iASPP.

In the present study, the function of miR-150 and its downstream target iASPP in the growth and metastasis of colorectal cancer (CRC) cells was investigated. The expression of miR-150 and iASPP was first investigated in clinical CRC samples. Subsequently, the effects of miR-150 overexpression and iASPP inhibition on cell viability, cell cycle distribution, apoptosis, migration and invasion were detected with CCK-8, flow cytometry, scratch and Transwell assays. The interaction between miR-150 and iASPP was confirmed using a dual-luciferase assay. Subsequently, the key role of iASPP in the anti-CRC function of miR-150 was assessed by inducing the expression of the gene in miR-150 overexpressed SW480 cells. In clinical samples, the level of miR-150 was downregulated, while iASPP was induced. Enforced expression of miR-150 decreased the viability, induced G1 cell cycle arrest and apoptosis, and inhibited the migration and invasion of SW480 cells. Knockdown of iASPP exerted a similar effect on SW480 cells to that of the overexpression of miR-150. Dual-luciferase assay demonstrated that miR-150 directly bound to iASPP and inhibited its transcription. The function of miR-150 depended on the inhibition of iASPP; induced expression of iASPP in miR-150-knockdown SW480 and HCT116 cells restored cell viability, migration and invasion while inhibiting G1 cell cycle arrest and apoptosis. Increased expression of miR-150 suppressed viability, proliferation, migration and invasion of SW480 cells. Furthermore, iASPP was a direct target of miR-150 and played a key role in its anti-CRC function. miR-150 may be a promising predictor of prognosis in CRC patients.

[Application of multidisciplinary treatment in patients with liver metastasis of colorectal cancer].

OBJECTIVE: To evaluate the application of multidisciplinary treatment (MDT) in patients with liver metastasis of colorectal cancer(CLM). METHODS: Clinical data of 118 patients with liver metastasis of colorectal cancer, including 32 patients with MDT (MDT group) and 86 patients without MDT (control group), from February 2014 to April 2015 in PLA General Hospital were analyzed retrospectively. Compliance of preoperative examination and adjuvant therapy, and efficacy-associated indexes were compared between the two groups. RESULTS: (1) As compared to control group, statistically significant increase in imaging examination ratio was found in MDT group: chest CT [87.5%(28/32) vs. 40.7%(35/86), P=0.0000], abdominal MRI [84.4%(27/32) vs.61.6%(53/86), P=0.019], pelvic MRI [63.7%(7/11) vs. 24.3%(8/33), P=0.017]. The preoperative assessment of TNM staging was also higher in MDT group [100%(32/32) vs. 20.9%(18/86), P=0.0000], while there was no significant difference in accuracy rate of TNM staging between the two groups [81.3%(26/32) vs. 66.7%(12/18), P=0.2465]. (2) Rates of preoperative chemotherapy and chemotherapy completion were also higher in MDT group than those in control group [90.6%(29/32) vs. 62.8%(54/86), P=0.0033; 82.8% (24/29) vs. 57.4% (31/54), P=0.000], but conversion rate of unresectable CLM showed no significant difference [24.0% (6/25) vs. 14.3% (7/49), P=0.299 ]. (3) Rate of one-stage resection or ablation was higher in MDT group compared to control group [76.9%(10/13) vs. 36.0%(9/25), P=0.038], and resection rate of metastasis nidus was also higher in MDT group [77.0%(20/26) vs. 44.9%(13/29), P=0.015]. No significant differences were observed in rates of R0 resection, positive surgical margin, lymph node clearance, ablation of metastasis nidus, pathological complete response, postoperative chemotherapy or postoperative complications (all P>0.05). CONCLUSION: MDT has the advantages on standardization of preoperative examination and perioperative chemotherapy, and can improve the rate of one-stage resection or ablation, as well as resection of metastasis nidus.

RNF43 Inhibits Cancer Cell Proliferation and Could be a Potential Prognostic Factor for Human Gastric Carcinoma.

BACKGROUND/AIMS: RNF43 is a member of transmembrane E3 ubiquitin ligases and plays important roles in tumor formation progression. In current study, we aimed to explore RNF43 expression and analyze its role in gastric carcinoma. METHODS AND RESULTS: The level of RNF43 was detected in 77 cases of gastric carcinoma and matched normal tissues by real-time PCR, western blotting and immunohistochemistry. We found that the expression of RNF43 was significantly down-regulated in the gastric carcinoma tissues compared to the normal mucosae (all P<0.001). In addition, RNF43 was significantly correlated with histological differentiation (P = 0.001), T-stage cancer (P<0.001), depth of invasion (P<0.001), metastasis of regional lymph nodes (P<0.001), pTNM stage (P<0.001) and survival (P = 0.021). We further explored the biological functions of RNF43 in gastric carcinoma cell lines. Both gain- and loss-function assays show that RNF43 could suppress cell proliferation while promotes cell apoptosis. Further, we found that RNF43 was positively correlated with p53 and cleaved-caspase3 and negatively correlated with Ki67 and Lgr5. CONCOLUSION: In conclusion, RNF43 might act as a tumor suppressor in gastric carcinoma and might be a potential indicator for the clinical assessment of gastric cancer prognosis.

Clinical significance of immunotherapy with combined three kinds of cells for operable colorectal cancer.

Surgery, chemotherapy, and radiotherapy have presented with the ability of killing tumor cells, as well as damaging the immune function, which can be corrected by the immunotherapy. The purpose of this perspective cohort study was to evaluate the efficacy of postoperative immunotherapies of tumor lysate-loaded dendritic cells (DC), in vitro DC-activated T (DC-AT), and activated T cells (ATC) combined with chemotherapy on the survival of patients with operable colorectal cancer. A total of 253 patients with primary colorectal cancer resection including 181 patients receiving postoperative simple chemotherapy (control group) and 72 patients receiving immunotherapies of DC, DC-AT, and ATC combined with chemotherapy during the corresponding period (immunotherapy group) were enrolled in this perspective cohort study. The survival of these patients was analyzed. The immunotherapy group presented a higher 5-year overall survival rate than the control group (75.63 vs 67.81 %, P = 0.035), as well as 3-year overall survival rate (87.07 vs 74.80 %, P = 0.045). For patients with advanced cancer (TNM stages III and IV), immunotherapy significantly promotes mean survival than control subjects (59.74 ± 3.21 vs 49.99 ± 2.54 years, P = 0.034). Patients who received more than three cycles of immunotherapies had a higher 5-year overall survival rate than those with less than three cycles (82.10 vs 69.90 %, P = 0.035). No serious adverse effect was observed in the immunotherapy group. Postoperative immunotherapies with DC, DC-AT, and ATC combination can promote the survival of patients with operable colorectal cancer (Clinical Trials, ChiCTR-OCH-12002610).

Berberine suppresses in vitro migration of human aortic smooth muscle cells through the inhibitions of MMP-2/9, u-PA, AP-1, and NF-κB.

Berberine, a type of isoquinoline alkaloid isolated from Chinese medicinal herbs, has been reported to have various pharmacological activities. Studies have demonstrated that berberine has beneficial effects on vascular remodeling and alleviates restenosis after vascular injury. However, its mechanism of action on vascular smooth muscle cell migration is not fully understood. We therefore investigated the effect of berberine on human aortic smooth muscle cell (HASMC) migration. Boyden chamber assay was performed to show that berberine inhibited HASMC migration dosedependently. Real-time PCR and Western blotting analyses showed that levels of matrix metalloproteinase (MMP)-2, MMP-9, and urokinase-type plasminogen activator (u-PA) were reduced by berberine at both the mRNA and protein levels. Western blotting assay further confirmed that activities of c-Fos, c-Jun, and NF-κB were significantly attenuated. These results suggest that berberine effectively inhibited HASMC migration, possibly by down-regulating MMP-2, MMP-9, and u-PA; and interrupting AP-1 and NF-κB mediated signaling pathways.

[Application of laparoscopic technique in acute abdomen of gastrointestinal surgery].

OBJECTIVE: To explore the clinical application of laparoscopy in gastrointestinal abdominal emergency. METHODS: Clinical data of 44 cases with undefined acute abdomen undergoing laparoscopic surgery from October 2008 to October 2011 were analyzed retrospectively. Sixty-five cases treated by regular surgery during the same period were enrolled as controls. RESULTS: In laparoscopic surgery group, 42 cases were diagnosed under laparoscopy(95.5%, 42/44). Thirty-four (77.3%,34/44) patients received operation successfully after diagnosis, including 20 of total laparoscopy, 14 of assistant small incision. Compared with control group, laparoscopic group had shorter length of incision[(6.7±2.2) cm vs. (15.8±3.4) cm], less blood loss[(51.4±30.3) ml vs. (117.9±49.5) ml], faster recovery of postoperative gastrointestinal function[postoperative oral intake(15.0±6.1) d vs. (30.5±8.4) d], shorter hospital stay[(5.6±4.2) d vs. (8.4±4.8) d] (all P<0.05), lower complication rate, and less surgical cost(P>0.05). CONCLUSION: Laparoscopy is safe and effective in treating gastrointestinal abdominal emergency and therapeutic operation can be performed after a definite diagnosis.

Dynamic tracing of immune cells in an orthotopic gastric carcinoma mouse model using near-infrared fluorescence live imaging.

Adoptive cellular immunotherapy (ACI) has been demonstrated to be a promising cancer therapeutic, however, the distribution of immune cells injected into a tumor-bearing body is unclear. In this study, we investigated the tumor-targeting capacity of cytokine-induced killer (CIK) cells and cytotoxic T lymphocytes (CTLs) in a human gastric carcinoma orthotopic mouse model using a near-infrared fluorescence imaging system. CIK cells and tumor-specific CTLs were prepared with the near-infrared fluorescent dye DiR. As expected, no significant change in the proliferation rate or antitumor activity of CIK cells and CTLs was noted after labeling with DiR. Furthermore, a gastric carcinoma orthotopic model was established using a fibrinogen-thrombin method in nude mice followed by intraperitoneal infusion of the labeled immune cells into nude mice with established gastric carcinoma. Dynamic tracing of the immune cells was performed using a fluorescence-based live imaging system. Concentrated fluorescence signals were observed for a minimum of two weeks at the tumor site in mice infused with either CIK cells or CTLs with a peak signal at 48 h. Notably, CTLs were more persistent at the tumor site and exhibited a more intense antitumor activity than CIK cells following infusion. These results provided visual evidence of the tumor-targeting capacity of immune cells in live animals.

[Diagnosis and surgical therapy of primary retroperitoneal neurogenic tumors].

OBJECTIVE: To explore the diagnosis and surgical therapy for retroperitoneal neurogenic tumors (PRNTs). METHODS: The clinical records of 79 surgically treated patients with retroperitoneal neurogenic tumor were retrospectively analyzed. RESULTS: Twenty-nine patients presented with abdominal pain, 26 with abdominal mass, 15 with inferior extremities pain and numbness and 9 patients without clinical symptoms. Type B ultrasound (BUS), CT scan and surgical resection were performed for all the patients. Pathological analysis identified 19 patients with neurofibroma, 8 with neurilemmoma, 4 with paraganglioma, 21 with neurofibrosarcoma, 14 with malignant neurilemmoma, 6 with malignant paraganglioma, 5 with neuroectodermal tumor and 2 with neuroblastoma. The mortality rate of PRNT operation is was 1.3%, 3-year recurrence rate of benign tumor 0%, 5-year recurrence rate of benign tumor 12.9%, reoperation rate 100%, 5-year survival rate 100%. 3 years recurrence rate of malignancy tumor 41.6%, reoperation rate 90%, 5 years recurrence rate of malignancy tumor 79.1% and 5 years survival rate is 62.5%. CONCLUSION: BUS, CT and MRI are decisive for localization diagnosis. Surgical resection is the mainstay of therapy for this disease. Pre-operative preparation of intestinal tract and blood, maintaining the intactness of involved nerve are important for tumor resection. To prevent tumor recurrence, the key surgical techniques are to minimize tumor residues and . to handle intervertebral foramen properly.

Absolute quantification of plasmid DNA by real-time PCR with genomic DNA as external standard and its application to a biodistribution study of an HIV DNA vaccine.

Real-time quantitative PCR (QPCR) has been proven to be a powerful tool for quantifying specific target DNA sequences. Compared to relative quantification, absolute quantification has the advantage of determining the absolute copy number of a given target, such as pathogen or plasmid DNA in vivo. However, matrix or impurities remaining in a DNA sample after various sample treatment procedures may influence a subsequent DNA analysis. In this work, we have compared methods of sample processing and validated the use of tissue genomic DNA as a universal external standard to facilitate quantification of plasmid DNA in biological matrix, especially addressing the amplification inhibition due to matrix effect and sample complexity. Also, we applied our high-throughput sample preparation and absolute quantification method to determine the distribution of an HIV plasmid DNA vaccine in vivo. Successful application showed the validity and reliability of the method in absolute quantification of a particular gene in vivo.