Artificial intelligence system of faster region-based convolutional neural network surpassing senior radiologists in evaluation of metastatic lymph nodes of rectal cancer.

BACKGROUND: An artificial intelligence system of Faster Region-based Convolutional Neural Network (Faster R-CNN) is newly developed for the diagnosis of metastatic lymph node (LN) in rectal cancer patients. The primary objective of this study was to comprehensively verify its accuracy in clinical use. METHODS: Four hundred fourteen patients with rectal cancer discharged between January 2013 and March 2015 were collected from 6 clinical centers, and the magnetic resonance imaging data for pelvic metastatic LNs of each patient was identified by Faster R-CNN. Faster R-CNN based diagnoses were compared with radiologist based diagnoses and pathologist based diagnoses for methodological verification, using correlation analyses and consistency check. For clinical verification, the patients were retrospectively followed up by telephone for 36 months, with post-operative recurrence of rectal cancer as a clinical outcome; recurrence-free survivals of the patients were compared among different diagnostic groups, by methods of Kaplan-Meier and Cox hazards regression model. RESULTS: Significant correlations were observed between any 2 factors among the numbers of metastatic LNs separately diagnosed by radiologists, Faster R-CNN and pathologists, as evidenced by rradiologist-Faster R-CNN of 0.912, rPathologist-radiologist of 0.134, and rPathologist-Faster R-CNN of 0.448 respectively. The value of kappa coefficient in N staging between Faster R-CNN and pathologists was 0.573, and this value between radiologists and pathologists was 0.473. The 3 groups of Faster R-CNN, radiologists and pathologists showed no significant differences in the recurrence-free survival time for stage N0 and N1 patients, but significant differences were found for stage N2 patients. CONCLUSION: Faster R-CNN surpasses radiologists in the evaluation of pelvic metastatic LNs of rectal cancer, but is not on par with pathologists. TRIAL REGISTRATION: www.chictr.org.cn (No. ChiCTR-DDD-17013842).

Clinicopathological and prognostic significance of aberrant Arpin expression in gastric cancer.

AIM: To detect the expression of Arpin, and determine its correlation with clinicopathological characteristics and the prognosis of gastric cancer (GC) patients. METHODS: A total of 176 GC patients were enrolled as study subjects and classified into groups according to different clinicopathological variables. GC mucosal tissues were obtained via surgery. Another 43 paraffin-embedded tissue blocks of normal gastric epithelium (> 5 cm away from the edge of the tumor) were included in the control group. Immunohistochemistry (IHC) for the Arpin and Arp3 proteins was performed on the formalin-fixed, paraffin-embedded GC tissues. Additionally, expression of the Arpin protein in 43 normal gastric tissues was also determined using IHC. RESULTS: Expression of the Arpin protein in GC was lower than that in normal gastric mucosa (30.68% vs 60.47%, P < 0.001). A χ2 test of the 176 GC samples used for IHC showed that decreased Arpin expression was associated with advanced TNM stage (P < 0.01) and the presence or absence of lymph node metastasis (80.92% vs 35.56%, P < 0.001). Additionally, a significant correlation was observed between the expression of Arpin and the presence of the Arp2/3 complex in GC tissues (χ2 = 30.535, P < 0.001). Moreover, a multivariate Cox regression analysis revealed that Arpin expression [hazard ratio (HR) = 0.551, P = 0.029] and TNM stage (HR = 5.344, P = 0.001) were independent prognostic markers for overall survival of GC patients. Regarding the 3-year disease-free survival (DFS), the recurrence rate of GC patients with low Arpin expression levels (median DFS 19 mo) was higher than that in the high-Arpin-expression group (median DFS 34 mo, P = 0.022). CONCLUSION: Low Arpin levels are associated with clinicopathological variables and a poor prognosis in GC patients. Arpin may be regarded as a potential prognostic indicator in GC.

[Regulation of p14(ARF) expression and induction of cell apoptosis with c-myc in a p53-independent pathway].

OBJECTIVE: To explore the regulation of p14(ARF) expression and induction of cell apoptosis with the mutant and wild-type c-myc genes in a p53-independent pathway of signal transduction. METHODS: The mutant and wild-type c-myc genes were transfected by lentivirus into HCC1937 to form the stable over-expression cell lines. Uninfected cells and lentivirus-infected ones carrying no c-myc gene acted as blank and infection controls respectively. And c-myc and p14(ARF) mRNA and protein, proliferation and apoptosis in HCC1937 with mutant and wild-type c-myc were detected by reverse transcription (RT)-PCR, Western blotting, thiazolyl blue tetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase mediated X-dUTP nick end labeling (TUNEL) respectively. RESULTS: After the lentivirus-mediated gene transfer, c-myc mRNA and protein expression increased in the mutant and wild-type groups. p14(ARF) mRNA and protein increased in the wild-type group and the mutant group and there were significant difference between them with blank and infection controls (mutant groups: 0.560 ± 0.010, 0.154 ± 0.011, wild-type groups: 0.651 ± 0.010, 0.382 ± 0.013, both P < 0.05). The group of mutant and wild-type c-myc could promote the proliferation of cell growth. And c-myc was more effective to induce apoptosis in the wild-type group as compared with the mutant group (7.1% ± 0.7% vs 3.2% ± 0.4%, P < 0.05). CONCLUSIONS: In a p53-independent pathway, the over-expression of wild-type c-myc obviously up-regulates the expression of p14(ARF). And cell apoptosis may be induced through the regulation of p14(ARF)-related gene, keep balance of proliferative promotion and apoptosis induction. When there is a loss-of-function of mutant c-myc, tumorigenicity increases via a disturbed balance of proliferative promotion and apoptosis induction.

[Effect of early oral enteral nutrition on clinical outcomes after gastric cancer surgery].

OBJECTIVE: To investigate the effect of early oral feeding with enteral nutrition preparation after surgery on clinical outcomes in patients with gastric cancer. METHODS: Sixty patients with gastric cancer undergoing radical operation between July 2010 and May 2011 were randomly divided into two groups using random digit table: experimental group(n=30, administration of water and enteral nutrition early after surgery) and control group(n=30, conventional postoperative care protocol). Clinical outcomes, immune function, and nutritional status between the two groups were compared. RESULTS: As compared to the control group, duration of fever was significantly shorter in the experimental group [(81.1±6.4) h vs. (87.3±8.0) h, P<0.05], as were postoperative time of flatus [(79.9±9.5) h vs. (86.6±8.7) h, P<0.05] and postoperative hospital stay [(7.83±2.23) d vs. (9.57±1.96) d, P<0.01]. The medical cost [(30,220±3,220) RMB vs.(34,600±32,120) RMB, P<0.01] was lower than that in the control group. There was no significant difference in morbidity between the two groups[13.3%(4/30) vs. 16.7%(5/30), P>0.05]. The levels of CD3(+)T, CD4(+)T, NK cell, CD4(+)T/CD8(+)T, albumin, and prealbumin were higher in the experimental group as compared to the control group on postoperative day 3 and 7(P<0.05). CONCLUSION: Early oral feeding with enteral nutrition preparation after surgery can improve the nutritional status and immune function, and accelerate the rehabilitation for patients with gastric cancer.

Clinicopathologic significance of slug expression in human intrahepatic cholangiocarcinoma.

AIM: To explore the expression and function of slug, a transcriptional repressor, in human intrahepatic cholangiocarcinoma (IHCC) and identify its role in IHCC progression. METHODS: Expression of slug was detected in 36 cases of IHCC and 12 cases of normal intrahepatic bile ducts and liver parenchyma by immunohistochemistry. The patients were divided into low slug expression group (< 20% of carcinoma cells stained) and high slug expression group (> or = 20% of carcinoma cells stained). Slug expression was correlated with clinicopathological parameters of IHCC patients. The patients were defined as short-term survivors if their survival time was < 12 mo and as long-term survivors if their survival time was > or = 12 mo. RESULTS: Slug was not expressed in normal liver epithelium samples, lowly expressed in 15 tissue samples (10 -, 5 +) and highly expressed in 21 tissue samples (16 ++; 5 +++) from IHCC patients. The survival rate of patients with a low slug expression was 33.3% (n = 5) and 66.7% (n = 10), respectively. The survival rate of patients with a high slug expression was 61.9% (n = 13) and 38.1% (n = 8), respectively (P = 0.02). Lymph node metastasis was found in 4 (26.7%) out of the 15 patients with a low slug expression and in 14 (66.7%) out of the 21 patients with a high slug expression, respectively. The incidence rate of lymph node metastasis increased with the increasing slug expression level (P = 0.003), and higher in patients with a high slug expression than in those with a low slug expression. Slug expression did not significantly correlate with the tumor size and stage or histologic grade, or with the gender and age of patients CONCLUSION: Slug expression is a novel prognostic marker for IHCC with lymph node metastasis.

[Association of interleukin-10 gene polymorphism with cachexia in patients with gastric cancer].

OBJECTIVE: To investigate whether the single nucleotide polymorphisms (SNPs) at -1082, -819 and -592 of interleukin-10 gene and its haplotype are associated with cachexia in patients with gastric cancer. METHODS: Radioimmunoassay was used to examine the serum levels of IL-10 in 223 patients with gastric cancer. The single nucleotide polymorphisms (SNPs) of IL-10 gene -1082G/A, -819T/C and -592A/C were analyzed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The serum levels of IL-10 were significantly higher in patients with cachexia than those without (P < 0.001). An increased frequency of -1082G allele was noted in patients with cachexia (P = 0.049). The frequencies of -1082AG and -819CC genotypes were elevated in patients with cachexia than those without (P = 0.036, 0.024). In a logistic regression analysis adjusted for actual weight, carcinoma location and stage, the -1082AG genotype was associated with an odds ratio of 1.989 (95%CI, 1.041 - 3.802, P = 0.037), and the -819CC genotype with an odds ratio of 3.393 (95%CI, 1.298 - 8.871, P = 0.013) for cachexia. Furthermore, haplotype analysis revealed that G1082C819C592 haplotype was associated with a significantly increased risk of cachexia (OR = 2.21; 95%CI, 1.14 - 4.30; P = 0.02). CONCLUSION: Our results suggest that the gene haplotype of IL-10 contributes to the occurrence of cachexia in patients with gastric cancer in Chinese population.

[Association of cytokine gene polymorphisms with cachexia related to sporadic female breast cancer].

OBJECTIVE: To investigate the relation of the frequency of tumor necrosis factor (TNF)-alpha - 308, TNF-beta + 252, IL-1 beta + 3954, and IL-10 - 1082 gene polymorphisms to female breast cancer. METHODS: Peripheral blood samples were collected from 102 breast cancer patients with cachexia and 120 breast cancer patients without cachexia. Biallelic polymorphisms were performed by analyzing the incision enzyme-digested DNA fragment obtained using PCR. RESULTS: The allele frequencies of TNF-beta + 252, IL-1 beta + 3954, and IL-10 -1082 in the patients with cachexia were comparable with those of the patients without cachexia (all P > 0.05). The patients with cachexia showed a significantly higher prevalence of TNF2 than the patients without cachexia (20.6 % vs 10.0 %, P = 0.027). Logistic regression analysis indicated TNF2 as a risk factor for cachexia in breast cancer ( OR = 2.333, 95% CI: 1.085-5.017). CONCLUSION: TNF2 plays an important role in the susceptibility of cachexia in breast cancer.

Relationship between matrix metalloproteinase 2 and lung cancer progression.

BACKGROUND AND OBJECTIVE: Matrix metalloproteinases (MMPs) play an important role in several steps of cancer development. MMP2 and MMP9 have previously been implicated in lymphatic and vascular invasion of lung cancer; however, the expression and prognostic significance of MMP2 and MMP9 is not fully clarified. This study was designed to investigate the significance of MMP2 and MMP9 in lung cancer tissue or serum, and their correlation with lung cancer prognosis. METHODS: Immunohistochemical analysis was performed to determine MMP2 and MMP9 staining in human nonsmall cell lung cancer (NSCLC). Serum MMP2 and MMP9 protein levels in patients after surgery were measured using the ELISA method. The correlation between MMP2 and MMP9 serum levels and clinicopathological features of NSCLC were analyzed by survival analysis. We also performed reverse transcriptase (RT)-PCR assays to detect messenger RNA (mRNA) expression to further confirm the activity of MMP2 and MMP9 in human lung cancer. RESULTS: Increased MMP2 immunostaining and MMP2 serum level correlated with advanced tumor stage and the presence of distant metastasis (Pearson's chi(2) test and ANOVA, p < 0.05). However, for MMP9, only serum level showed a correlation with advanced tumor stage. No significant correlation was observed between MMP2 or MMP9 immunostaining expression and tumor histologic features (Pearson's chi(2) test, p = 0.061 and p = 0.087, respectively). A high densitometry value of MMP2 and MMP9 PCR products (i.e. mRNA expression level) was related to poor differentiation grade, distant metastasis, and small cell carcinoma histologic type (ANOVA, p < 0.05). CONCLUSIONS: Our results suggest that MMP2 is a more sensitive predictor than MMP9 of lung cancer progression, metastasis, and survival. Serum MMP2 levels may be a valuable prognosis variable and could help to stratify lung cancer patients into low- and high-risk groups.

Effects of L-arginine on serum nitric oxide, nitric oxide synthase and mucosal Na+-K+-ATPase and nitric oxide synthase activity in segmental small-bowel autotransplantation model.

AIM: To explore a simple method to create intestinal autotransplantation in rats and growing pigs and to investigate the effect of L-arginine supplementation on serum nitric oxide (NO), nitric oxide synthase (NOS) and intestinal mucosal NOS and Na+-K+-ATPase activity during cold ischemia-reperfusion (IR) in growing pigs. METHODS: In adult Wistar rat models of small bowel autotransplantation, a fine tube was inserted into mesenteric artery via the abdominal aorta. The superior mesenteric artery and vein were occluded. Isolated terminal ileum segment was irrigated with Ringer's solution at 4 degrees and preserved in the same solution at 0-4 degrees for 60 min. Then, the tube was removed and reperfusion was established. In growing pig models, a terminal ileum segment, 50 cm in length, was isolated and its mesenteric artery was irrigated via a needle with lactated Ringer's solution at 4 degrees. The method and period of cold preservation and reperfusion were described above. Ten white outbred pigs were randomly divided into control group and experimental group. L-arginine (150 mg/kg) was continuously infused for 15 min before reperfusion and for 30 min after reperfusion in the experimental group. One, 24, 48, and 72 h after reperfusion, peripheral vein blood was respectively collected for NO and NOS determination. At the same time point, intestinal mucosae were also obtained for NOS and Na+-K+-ATPase activity measurement. RESULTS: In adult rat models, 16 of 20 rats sustained the procedure, three died of hemorrhage shock and one of deep anesthesia. In growing pig models, the viability of small bowel graft remained for 72 h after cold IR in eight of 10 pigs. In experimental group, serum NO level at 1 and 24 h after reperfusion increased significantly when compared with control group at the same time point (152.2+/-61.4 micromol/L vs 60.8+/-31.6 micromol/L, t=2.802, P=0.02<0.05; 82.2+/-24.0 micromol/L vs 54.0+/-24.3 micromol/L, t=2.490, P=0.04<0.05). Serum NO level increased significantly at 1 h post-reperfusion when compared with the same group before cold IR, 24 and 48 h post-reperfusion (152.2+/-61.4 micromol/L vs 75.6+/-16.2 micromol/L, t=2.820, P=0.02<0.05, 82.2+/-24.0 micromol/L, t=2.760, P=0.03<0.05, 74.2+/-21.9 micromol/L, t=2.822, P=0.02<0.05). Serum NOS activity at each time point had no significant difference between two groups. In experimental group, intestinal mucosal NOS activity at 1 h post-reperfusion reduced significantly when compared with pre-cold IR (0.79+/-0.04 U/mg vs 0.46+/-0.12 U/mg, t=3.460, P=0.009<0.01). Mucosal NOS activity at 24, 48, and 72 h post-reperfusion also reduced significantly when compared with pre-cold IR (0.79+/-0.04 U/mg vs 0.57+/-0.14 U/mg, t=2.380, P=0.04<0.05, 0.61+/-0.11 U/mg, t=2.309, P=0.04<0.05, 0.63+/-0.12 U/mg, t=2.307, P=0.04<0.05). In control group, mucosal NOS activity at 1 and 24 h post-reperfusion was significantly lower than that in pre-cold IR (0.72+/-0.12 U/mg vs 0.60+/-0.07 U/mg, t=2.320, P=0.04<0.05, 0.58+/-0.18 U/mg, t=2.310, P=0.04<0.05). When compared to the normal value, Na+-K+-ATPase activity increased significantly at 48 and 72 h post-reperfusion in experimental group (2.48+/-0.59 micromol/mg vs 3.89+/-1.43 micromol/mg, t=3.202, P=0.04<0.05, 3.96+/-0.86 micromol/mg, t=3.401, P=0.009<0.01) and control group (2.48+/-0.59 micromol/mg vs 3.58+/-0.76 micromol/mg, t=2.489, P=0.04<0.05, 3.67+/-0.81 micromol/mg, t=2.542, P=0.03<0.05). CONCLUSION: This novel technique for intestinal autotransplantation provides a potentially consistent and practical model for experimental studies of graft cold preservation. L-arginine supplementation during cold IR may act as a useful adjunct to preserve the grafted intestine.

Association of two polymorphisms of tumor necrosis factor gene with acute biliary pancreatitis.

AIM: To investigate TNF-alpha-308 and TNFB polymorphisms in acute biliary pancreatitis (ABP) and to related them to the plasma TNF-alpha levels. METHODS: Genomic DNA was prepared from peripheral blood leukocytes. Genotypes and allele frequencies were determined in patients (n=127) and healthy controls (n=102) using restriction fragment length polymorphism analysis of polymerase chain reaction (PCR) products. Reading the size of digested bands from polyacrylamide gel demonstrated the two alleles TNF1 and TNF2, or the two alleles TNFB1 and TNFB2. RESULTS: The frequencies of TNF2 polymorphism and TNFB2 polymorphism were both similar in patients with mild or severe pancreatitis, so were in pancreatitis patients and in controls. Patients with septic shock showed a significantly higher prevalence of the TNF2 than those without. No significant differences were found in the genotype distribution of TNF-alpha-308 and TNFB among different groups. Plasma TNF-alpha levels did not differ significantly in ASBP patients displaying different alleles of the TNF gene studied. CONCLUSION: Results indicate that TNF gene polymorphisms studied play no part in determination of disease severity or susceptibility to acute biliary pancreatitis; however, TNF2 polymorphism is associated with septic shock from ASBP. Genetic factors are not important in determining plasma TNF-alpha levels in ASBP.

[The associations of the single nucleotide polymorphisms on TNF and CD14 promoters with the mortality of infection, systematic inflammatory response syndromec and sepsis in surgical patients].

OBJECTIVE: To investigate the association of Single nucleotide polymorphism (SNPs) tumor necrosis factor (TNF) and CD14 promoter with the systematic inflammatory response syndromec (SIRS) and sepsis in surgical patients. METHODS: The DNA and RNA sample of PBMC from 113 patients, 40 of them being complicated with sepsis, and 100 healthy volunteers were extracted. The SNP genotypes of TNF-alpha -308 G/A, -863 C/A, CD14-159C/T and TNFB1/B2 were examined by restriction fragment length polymorphism PCR (PCR-RFLP). The expressions of TNF-alpha mRNA of PBMC in parts of the patients who have at least one genotype of SNP were detected by RT-PCR. The risks for sepsis associated with polymorphisms in the TNF-alpha or CD14 promoter were determined by multivariate analysis. RESULTS: The rates of TNF2, -863A, CD14-159T alleles were 15%, 32.5%, and 40% respectively in patients with sepsis, significantly higher than those in the patients with SIRS (8.9%, 22%, and 23.3%), and those in the healthy volunteers (5%, 16% and 26%). The expression of TNF-alpha mRNA was much higher in those patients with at least one kind of SNP than those without SNP. CONCLUSION: The A-allele at the -308 and -863 position in the TNF-alpha promoter and the T-allele at the -159 position in the CD14 promoter increase the risk for sepsis. The effect of SNP genotypes on TNF-alpha expression can modulate inflammatory response.