Matrix metalloproteinase-2 inducing COL1A1 synthesis via integrin alpha â ¤ promotes invasion and metastasis of cholangiocarcinoma cells.

INTRODUCTION AND OBJECTIVES: Cholangiocarcinoma (CCA) is characterized by early distant invasion and metastasis, whereas the underlying mechanism is still obscure. Increasing evidence shows that collagen type Ι alpha 1 (COL1A1) is a gene associated with the progression of multiple diseases. Here, we attempted to investigate the role of COL1A1 in CCA. MATERIALS AND METHODS: The expression of COL1A1 between tumor tissues and adjacent normal tissues obtained from CCA patients was detected by Western blot and immunofluorescence, followed by analysis of its clinical significance. Then, the biological effects of COL1A1 overexpression or knockdown on CCA cells were evaluated in vitro and in vivo. Finally, molecular mechanism of COL1A1 in regulating the invasion and metastasis of CCA cells was determined by a series of experiments. RESULTS: COL1A1 expression was significantly higher in CCA pathological tissues than in corresponding adjacent normal tissues. Analysis of 83 CCA patients showed that higher expression of COL1A1 was correlated with poorer patient prognosis. Notably, overexpression or knockdown experiments revealed that COL1A1 contributed to the migration and invasion, as well as epithelial-to-mesenchymal transition (EMT), in CCA cells. Further investigations demonstrated that matrix metalloproteinase-2 (MMP2) promoted COL1A1 upregulation via the integrin alpha Ⅴ pathway, therefore affecting ECM remodelling and inducing EMT in CCA cells. Moreover, COL1A1 expression was positively related to PD-1 and PD-L1 in CCA, and COL1A1 increased PD-L1 expression by activating the NF-κB pathway. CONCLUSIONS: COL1A1 plays an important role in regulating CCA progression and may act as a promising biomarker and therapeutic target for CCA.

Prealbumin and lymphocyte-based prognostic score, a new tool for predicting long-term survival after curative resection of stage II/III gastric cancer.

The aim of this retrospective study was to investigate the prognostic significance of pre-treatment immunological and nutritional statuses in patients with locally advanced gastric cancer (GC), and to use the risk factors to develop a predictive score. A total of 731 patients who underwent gastrectomy for stage II/III GC from November 2010 to December 2015 were recruited into this retrospective study. On the basis of univariate and further multivariate Cox regression analyses, decreased pretreatment lymphocyte count (<1·5×109/litre) and prealbumin concentrations (<180 mg/l) were identified to be independently associated with poorer overall survival (OS) and disease-free survival (DFS). Low albumin concentrations (<33 g/l) were identified as an independent risk factor only for OS, but not for DFS. Thereafter, patients who had a decreased prealbumin concentration and lymphocyte count were given a combination of serum prealbumin concentration and lymphocyte count (Co-PaL) score of 2. Patients with only one or neither of these concentrations were given a Co-PaL score of 1 or 0, respectively. Both the OS and the DFS time were inversely related to the Co-PaL scores, and the differences among the three groups were all significant. In contrast, the prognosis did not differ significantly between patients with good nutrition and those with mild to moderate malnutrition according to the prognostic nutritional index. This study indicated that the simple scoring system could accurately predict the prognosis of patients who underwent gastrectomy for stage II/III GC. The score might be helpful in terms of clinical preoperative decision-making.

[Multivariate analysis of risk factors for pulmonary infection after radical gastrectomy for gastric cancer].

OBJECTIVE: To explore the major risk factors for pulmonary infection after radical gastrectomy in patients with gastric cancer. METHODS: From November 2010 to February 2014, a total of 765 patients undergoing radical gastrectomy at our hospital were divided into 2 groups based upon the presence of postoperative pulmonary infection (n = 32, 4.2%) or not (n = 733, 95.8%). Their clinicopathological data were retrospectively analyzed by Logistic regressive analysis with a case-control study model. RESULTS: Comparing with the control group, the patients had longer surgical duration (245.7 ± 66.7 vs 210.9 ± 47.2 min, P < 0.01), higher rates of requiring intensive care (12.50% vs 2.86%, P = 0.02) and longer post-operative hospital stays (21.9 ± 24.9 vs 14.2 ± 4.2 days, P < 0.01) in the postoperative pulmonary infection group.Univariate Logistic regressive analysis found that age ≥ 60 years, smoking ≥ 400 year by cigarette, diabetes mellitus, chronic obstructive pulmonary disease, proximal or total gastrectomy, combined organ resection, surgical duration ≥ 240 min, intra-operative blood loss ≥ 300 ml, peri-operative transfusion, transfusion ≥ 3 unit packed red blood cell, post-operative transfusion and post-operative complications other than pulmonary infections were associated with postoperative pulmonary infection (all P < 0.05).Further multivariate analysis identified 4 independent risk factors for pulmonary infection after radical gastrectomy, including diabetes mellitus (OR = 4.77, 95%CI:1.18-19.23), post-operative complications other than pulmonary infections (OR = 3.15, 95%CI:1.25-7.90), intra-operative blood loss ≥ 300 ml (OR = 2.63, 95%CI:1.17-5.90) and post-operative nasogastric tube ≥ 5 days (OR = 2.30, 95%CI:1.02-5.21). CONCLUSION: Correcting the modifiable risk factors may reduce the incidence of pulmonary infection and shorten the length of hospital stays and costs after radical gastrectomy in patients with gastric cancer.

[Association of perioperative transfusion and postoperative complications after radical gastrectomy for gastric cancer].

OBJECTIVE: To explore the association of perioperative homologous blood transfusion (packed red blood cell, PRBC) and postoperative complications after radical gastrectomy in patients with gastric cancer. METHODS: From October 2010 to July 2013, a total of 636 patients undergoing radical gastrectomy at Department of Gastric, Duodenal & Pancreatic Surgery at Hunan Provincial Tumor Hospital were divided into 2 groups according to perioperative blood transfusion (n = 170, 26.73%) or not (n = 466, 73.27%). Their clinicopathological data, such as age, gender, co-morbidities, surgical duration, intraoperative blood loss volume and pathological stage were retrospectively analyzed by case-control study model. And the transfusion group was further divided into subgroup by transfusion volume (total PRBC<3.0, 3.0-7.5 or >7.5 U) and timing (pre-, intra- or post-operative) to examine the association of transfusion volume and timing with postoperative complications by Logistic regression. RESULTS: Thirty-two patients suffered from complications in the transfusion group (18.82%). And it was significantly more common than that in the control group (10.09% (47/466) , P < 0.01). Moreover, the complication rate (33.33% (12/36) ) was obviously higher in the large transfusion volume group (PRBC>7.5 U) than with those in the moderate (15.53% (16/103), P = 0.02) and low groups (12.90% (4/31) , P = 0.04). Infection was more common along with the total amount of transfused blood (6.45% (2/31), 10.68% (11/103) and 19.44% (7/36) in the low, moderate and large transfusion group respectively). Yet the differences were insignificant (P = 0.22). There was no significant difference of complication rates among the pre-, intra- and post-operative transfusion group classified by transfusion time (P = 0.39). And the postoperative infection rates were also insignificantly different (P = 0.88). Further Logistic analysis revealed that perioperative transfusion (OR = 2.71, 95% CI: 1.40-5.27, P < 0.01) was an independent risk factor for postoperative complications after radical gastrectomy. CONCLUSIONS: Perioperative blood transfusion is significantly associated with postoperative complications after radical gastrectomy in patients with gastric cancer. And a positive correlation exists between infection and the amount of transfused blood. But there was no association between transfusion time and complications. Thus decreasing perioperative transfusion may reduce the incidence of postoperative complications and shorten the length of hospital stays.

Comprehensive profiling of enhancer RNA in stage II/III colorectal cancer defines two prognostic subtypes with implications for immunotherapy.

BACKGROUND: Recently, enhancer RNAs (eRNAs) have garnered attention as pivotal biomarkers for the onset and progression of cancer. However, the landscape of eRNAs and the implications of eRNA-based molecular subtypes in stage II/III colorectal cancer (CRC) remain largely unexplored. METHODS: Comprehensive profiling of eRNAs was conducted on a public stage II/III CRC cohort with total RNA-seq data. We used unsupervised clustering of prognostic eRNAs to establish an eRNA-based subtyping system. Further evaluations included molecular characteristics, immune infiltration, clinical outcomes, and drug responses. Finally, we validated the eRNA-based subtyping system in The Cancer Genome Atlas (TCGA) CRC cohort. RESULTS: We identified a total of 6453 expressed eRNAs, among which 237 were prognostic. A global upregulation of eRNAs was observed in microsatellite-stable (MSS) CRCs when compared to microsatellite instability-high (MSI-H) CRCs. Through consensus clustering, two novel molecular subtypes, termed Cluster 1(C1) and Cluster 2(C2), were further identified. C1, associated with the activation of epithelial-mesenchymal transition (EMT), hypoxia, and KRAS signaling pathways, showed poorer prognosis. C2, correlated with the canonical CRC subtype, exhibited superior survival outcomes. In addition, C1 showed enrichment with immune infiltration and more sensitivity to immune checkpoint inhibitors. CONCLUSION: Our study unravels the molecular heterogeneity of stage II/III CRC at the eRNA level and highlights the potential applications of the novel eRNA-based subtyping system in predicting prognosis and guiding immunotherapy.

[Multivariate analysis of risk factors for intra-abdominal infections after radical gastrectomy for gastric cancer].

OBJECTIVE: To explore the major risk factors for intra-abdominal infections after radical gastrectomy in patients with gastric cancer. METHODS: From October 2010 to January 2013, a total of 479 patients undergoing radical gastrectomy at Department of Gastric, Duodenal & Pancreatic Surgery, Hunan Provincial Tumor Hospital were divided into 2 groups according to an onset of postoperative intra-abdominal infections (n = 32, 6.68%) or not (n = 447, 93.32%). Their clinicopathological data, such as age, gender, co-morbidities, surgical duration, operative blood loss and pathological stage were retrospectively analyzed by Logistic regressive analysis with a case-control study model. RESULTS: As compared with the control group, the patients had a greater age ((59 ± 10) vs (53 ± 11) years, P < 0.01), lower lymphocyte count ((1.4 ± 0.7) ×10(9)/L vs (1.7 ± 0.6) ×10(9)/L, P = 0.02), lower hemoglobin level ( (108 ± 28) vs (117 ± 24) g/L, P = 0.04), lower albumin level ((34 ± 6) vs (37 ± 5) g/L, P < 0.01) and longer surgical duration ((244 ± 43) vs (216 ± 45) min, P < 0.01) in the postoperative intra-abdominal infection group. Univariate Logistic regressive analysis found that a history of abdominal surgery, body mass index (BMI) >25 kg/m(2), co-morbidities, diabetes mellitus, complications due to gastric cancer, lymphocyte count <1.5×10(9)/L, hemoglobin <100 g/L, albumin <30 g/L, ascites, perioperative transfusion, total mastectomy, combined organ resection and surgical duration >240 min were associated with the occurrence of postoperative intra-abdominal infections (all P < 0.05). Further multivariate analysis identified 4 independent risk factors for intra-abdominal infections after radical gastrectomy, including combined multiorgan resection (OR = 3.64, 95%CI: 1.39-9.55), BMI>25 kg/m(2) (OR = 3.04, 95%CI: 1.17-7.92), diabetes mellitus (OR = 3.41, 95%CI: 1.05-11.09) and perioperative transfusion (OR = 2.24, 95%CI: 1.02-5.13). CONCLUSION: A correction of modifiable risk factors may reduce the incidence of intra-abdominal infections after radical gastrectomy, shorten the length of hospital stays and improve outcomes in patients with gastric cancer.

[Clavien-Dindo classification and risk factors for complications after radical gastrectomy for gastric cancer].

OBJECTIVE: To explore the complications after radical gastrectomy in patients with gastric cancer according to Clavien-Dindo classification and examine the major risk factors for complications. METHODS: From October 2010 to June 2013, a total of 614 patients undergoing radical gastrectomy at Department of Gastric,Duodenal & Pancreatic Surgery at Hunan Provincial Tumor Hospital were divided into 2 groups according to the occurrence of complications (n = 76, 12.38%) or not (n = 538, 87.62%). Their clinicopathological data, such as age, gender, co-morbidities, surgical duration, operative blood loss volume and pathological stage were retrospectively analyzed by Logistic regression with a case-control model. RESULTS: Among them, 76 patients developed complications (12.38%). According to Clavien-Dindo classification, 56(9.12%), 14(2.28%), 3(0.49%) and 3(0.49%) patients suffered stage II, III, IV and V complications respectively. Comparing with the control group, the patients had a higher transfusion rate (43.42% (n = 33) vs 24.16% (n = 130), P < 0.01) and a longer postoperative hospital stay in the complication group ((23 ± 18) vs (14 ± 6) days, P < 0.01). There was no difference in age, gender, body mass index (BMI), number of dissected lymph node, levels of hemoglobin and albumin at admission, intraoperative hemorrhage, surgical duration and pathological TNM stage in two groups (all P > 0.05). Univariate analysis revealed that BMI > 25 kg/m(2), co-morbidities, diabetes mellitus, complications due to gastric cancer, hemoglobin <100 g/L, albumin <30 g/L, ascites, total gastrectomy, combined multi-organ resection, surgical duration >240 min and perioperative transfusion were associated with postoperative complications (all P < 0.05).Further multivariate analysis showed that perioperative transfusion (OR = 2.78, 95%CI: 1.42-5.43, P < 0.01) and combined multi-organ resection (OR = 1.72, 95%CI: 1.14-2.58, P = 0.01) were independent risk factors for postoperative complications after radical gastrectomy. CONCLUSIONS: Classifying the complications after radical gastrectomy according to Clavien-Dindo classification is important for comparisons and quality assessments among different studies. And decreasing perioperative transfusion and avoiding combined multi-organ resection may reduce the incidence of postoperative complications and shorten the length of hospital stay.

Clinical value of the ThinPrep cytologic test with E6/E7 mRNA detection for cervical cancer screening in disease diagnosis.

BACKGROUND: To investigate the clinical value of the ThinPrep cytologic test (TCT) with the E6/E7 mRNA test for cervical cancer screening in disease diagnosis. METHODS: A total of 405 samples from Dazhou Central Hospital from April 2017 to July 2020 were collected, and we conducted a comparative analysis of the diagnostic performance of several test methods both individually andcombination. RESULTS: The sensitivity, specificity, positive predictive value, negative predictive value, accuracy, and area under the curve (AUC) were compared by single TCT, E6/E7 mRNA test, and combination methods. The TCT+E6/E7mRNA test was confirmed to have a relatively higher specificity of 80.32% (95% CI: 75.40%-84.48%, both P < 0.001), and AUC value (0.78, 95% CI: 0.73-0.83, and P < 0.001). CONCLUSION: The relative diagnostic value may be further improved by the combined detection of TCT and E6/E7 mRNA test. The combined detection of TCT and the E6/E7 mRNA test is expected to become a potential indicator for cervical lesions.

Platelet-derived PDGF promotes the invasion and metastasis of cholangiocarcinoma by upregulating MMP2/MMP9 expression and inducing EMT via the p38/MAPK signalling pathway.

Cholangiocarcinoma (CCA) is an aggressive tumour with a poor prognosis due to its late clinical presentation and the lack of effective non-surgical therapies. Previous studies have reported that platelets are implicated in tumour invasion and metastasis, while their role and the underlying mechanism in CCA remain unclear. Here, we show that platelets are hyperactivated in patients with CCA and that platelet-derived growth factor (PDGF) promotes the migration of CCA tumour cells both in vitro and in vivo. Further investigations revealed that PDGF can upregulate the expression of MMP2/MMP9 and induce epithelial-mesenchymal transition (EMT) by activating the p38/MAPK signalling pathway in CCA cells. In addition, the expression of MMP2/MMP9 was associated with lymph node metastasis and poor prognosis in CCA patients after surgical resection. In conclusion, our findings demonstrate that platelets play an important role in facilitating the invasion and metastasis of CCA cells by secreting PDGF, which may provide a novel target for CCA treatment.

S100A8 facilitates cholangiocarcinoma metastasis via upregulation of VEGF through TLR4/NF­κB pathway activation.

A growing body of evidence indicates that S100 calcium­binding protein A8 (S100A8) is frequently overexpressed in malignant tumor tissues and regulates tumor progression; however, the role of S100A8 in cholangiocarcinoma (CCA) remains unclear. The present study demonstrated that the protein expression of S100A8 was significantly higher in pathological tissues compared with adjacent normal tissues from patients with CCA. In addition, S100A8 expression was significantly associated with differentiation, lymph node metastasis and poor prognosis in patients following surgical resection of CCA. Furthermore, both in vitro and in vivo experiments revealed that overexpression of S100A6 promoted, while S100A8 knockdown attenuated, the migration and metastasis of CCA cells. Of note, the present results indicated that S100A8 promoted the CCA tumor cell­induced migration of vascular endothelial cells. Finally, S100A8 was demonstrated to positively regulate the expression of vascular endothelial growth factor (VEGF) in CCA cells, which was mediated by activation of the Toll­like receptor 4 (TLR4)/NF­κB pathway. In conclusion, the present study demonstrated that S100A8 had an important role in facilitating CCA cell migration and metastasis via upregulation of VEGF expression by activating the TLR4/NF­κB pathway. These findings may provide a novel target for CCA treatment.

[Corrigendum] S100A8 facilitates cholangiocarcinoma metastasis via upregulation of VEGF through TLR4/NF­κB pathway activation.

A growing body of evidence indicates that S100 calcium­binding protein A8 (S100A8) is frequently overexpressed in malignant tumor tissues and regulates tumor progression; however, the role of S100A8 in cholangiocarcinoma (CCA) remains unclear. The present study demonstrated that the protein expression of S100A8 was significantly higher in pathological tissues compared with adjacent normal tissues from patients with CCA. In addition, S100A8 expression was significantly associated with differentiation, lymph node metastasis and poor prognosis in patients following surgical resection of CCA. Furthermore, both in vitro and in vivo experiments revealed that overexpression of S100A8 promoted, while S100A8 knockdown attenuated, the migration and metastasis of CCA cells. Of note, the present results indicated that S100A8 promoted the CCA tumor cell­induced migration of vascular endothelial cells. Finally, S100A8 was demonstrated to positively regulate the expression of vascular endothelial growth factor (VEGF) in CCA cells, which was mediated by activation of the Toll­like receptor 4 (TLR4)/NF­κB pathway. In conclusion, the present study demonstrated that S100A8 had an important role in facilitating CCA cell migration and metastasis via upregulation of VEGF expression by activating the TLR4/NF­κB pathway. These findings may provide a novel target for CCA treatment.[the original article was published in International Journal of Oncology 56: 101­112, 2020; DOI: 10.3892/ijo.2019.4907].

Functional analysis of the correlation between ABCB11 gene mutation and primary intrahepatic stone.

The adenosine 5'­triphosphate binding cassette subfamily B member (ABCB)11 gene is involved in bile transport, and mutations in this gene are associated with cholestasis and cholelithiasis. Therefore, the aim of the present study was to investigate the association between ABCB11 gene mutation and primary intrahepatic stone (PIS)s and to investigate the mechanism through which ABCB11 gene mutations affect the expression of the corresponding protein. Mutations of the ABCB11 gene in 443 PIS patients and 560 healthy participants were detected by exon sequencing. The expression levels of ABCB11 mRNA and bile salt export pump (BSEP) protein in the liver tissues of patients with PISs were measured by quantitative polymerase chain reaction and western blot analysis. The mutant plasmids constructed by site­directed mutagenesis of the human BSEP gene were transfected into human embryonic kidney 293 (293) cells and Madin­Darby canine kidney (MDCK) cells, and the expression and distribution of rs118109635 of BSEP was measured. There were two significant mutations in the ABCB11 gene of the PIS patients compared with the healthy population; a missense mutation, rs118109635 (P=0.025), and a synonymous mutation, rs497692 (P=0.006). The two mutations were associated with the occurrence of preoperative jaundice (P=0.026, and P=0.011, respectively). The expression levels of BSEP in PIS patients with the missense mutation rs118109635 was decreased, whereas its mRNA expression levels remained unchanged. In PIS patients with the synonymous mutation rs497692, the expression levels of ABCB11 were decreased at both the mRNA and protein level. It was also found that mutation A865V reduced the expression levels of BSEP in 293 cells at the cellular level; its distribution in MDCK cell membranes was decreased, whereas its mRNA levels remained unchanged. The mutated loci at rs118109635 and rs497692 of the ABCB11 gene were correlated with PISs, causing a decreased expression of BSEP and reduced distribution of the protein in the cell membrane. Therefore, mutations at rs118109635 and rs497692 of the ABCB11 gene may be risk factors for PISs.

Incidence, causes and risk factors for 30-day readmission after radical gastrectomy for gastric cancer: a retrospective study of 2,023 patients.

The aim of this retrospective study was to investigate the incidence of, causes and risk factors for readmission to hospital ≤30 days after discharge of patients who underwent radical gastrectomy for gastric cancer. A total of 2,023 patients underwent radical gastrectomy operations from November 2010 to July 2017 in our hospital. Of these, 60 patients (3.0%) were readmitted within 30 days after their original discharge. The median time span between the index discharge and readmission was 14 days and the median time for readmission was 8 days. The main reasons for readmission were intestinal obstruction (n = 10, 16.7%), intra-abdominal fluid collection (n = 9, 15.0%), abdominal pain (n = 7, 11.7%), nutritional difficulty (n = 4, 6.7%) and anastomotic leakage (n = 4, 6.7%). Five patients (8.3%) required intensive care and 4 patients (6.7%) died from sudden cardiac arrest, gastrointestinal bleeding, sepsis or multiple organ dysfunctions. Multivariate analysis revealed that post-operative complications (Odds Ratio = 5.116, 95% confidence interval: 2.885-9.073, P < 0.001) was the only independent risk factor for readmission. Thus, appropriate strategies on discharge and close follow-ups for these high-risk patients should be drawn up in order to enhance significantly their quality of care.

Impact of peri-operative blood transfusion on post-operative infections after radical gastrectomy for gastric cancer: a propensity score matching analysis focusing on the timing, amount of transfusion and role of leukocyte depletion.

PURPOSE: Allogeneic blood transfusions (BTF) are sometimes inevitable during radical gastrectomy with lymphadenectomy for advanced gastric cancer. The aim of this retrospective study was to investigate the association between BTF and post-operative infections, focusing on the impact of timing, amount of transfusion and the role of leukocyte depletion. METHODS: The study cohort was 2064 patients who underwent gastrectomy for gastric cancer from November 2010 to August 2017. The association between BTF and post-operative infections was estimated by univariate and multivariate analyses after propensity score matching. Subgroup analysis was performed according to the timing and amount of transfusion, and leukocyte depletion or not. RESULTS: Out of a total 2064 patients, 426 (20.6%) received peri-operative BTF. After one-to-one matching, 361 pairs of patients were included for further analysis, of who 68 (9.4%) developed infections. Multivariate analysis identified that an operation time ≥ 240 min, combined multi-organ resection, BTF and BMI ≥ 25 kg/m2 were independent risk factors for post-operative infection. Patients given a high-volume (> 7.5 U), intra-operatively of leukocyte-non-depleted BTF had the highest risk of developing infections clarified by subgroup analysis. CONCLUSION: Infection was the most common complication following gastrectomy for gastric cancer and BTF was identified as an independent risk factor by propensity score matching and multivariate analyses. The timing, amount of transfusion and leukocyte depletion had an impact on the incidence of infection. To decrease infection, BTF should be avoided where possible, particularly during operation, with a large amount and leukocyte-not-depleted blood.

Intra-abdominal infection after radical gastrectomy for gastric cancer: Incidence, pathogens, risk factors and outcomes.

BACKGROUND: Surgical site infection, particularly intra-abdominal infection (IAI), remains a clinically important event after gastrectomy for gastric cancer. The aim of this retrospective study was to clarify the incidence, pathogens, risk factors and outcomes of IAI following gastrectomy for gastric cancer. METHODS: The study cohort was 1835 patients who underwent gastrectomy for gastric cancer from January 2011 through December 2016. The incidence, pathogens, and treatment outcomes of IAI were examined, and the risk factors were identified using univariate and multivariate analyses. RESULTS: In total, 73 patients (4.0%) developed IAI after radical gastrectomy. Bacterial culture in these patients showed that Gram-negative bacilli, such as Escherichia coli and Klebsiella pneumonia were the most common pathogens. Multivariate analysis identified that combined multi-organ resection (Odds Ratio [OR] = 2.262, 95% confidence interval [CI]: 1.114-4.596, P = 0.024), and body mass index (BMI) ≥ 25 kg/m2 (OR = 1.968, 95% CI: 1.107-3.500, P = 0.021) were independent risk factors. Three patients (4.1%) developed IAI who died from sepsis and/or multiple-organ failure, which was significantly higher than in the remaining 1762 patients without IAI (5 cases, 0.3%, P = 0.003). Moreover, IAI required more re-operations (5.5% vs 0.8%, P = 0.005) and longer post-operative hospital stays (23.3 days vs 11.2 days, P < 0.001) compared without IAI. CONCLUSIONS: IAI is a major complication after radical gastrectomy for gastric cancer, and associated with combined multi-organ resection and a BMI ≥ 25 kg/m2; thus, meticulous surgical procedures need to be performed in patients with these specific risk factors.

Variations of ABCB4 and ABCB11 genes are associated with primary intrahepatic stones.

Variations of the ABCB4 and ABCB11 genes affect the composition of bile and are associated with cholestasis and cholelithiasis. However, their roles in the formation of primary intrahepatic stones (PIS) remains to be elucidated. The aim of the present study was to determine whether there is an association between PIS and variations in these genes. Exon sequencing was performed in order to analyze the ABCB4 and ABCB11 genes of 176 patients with PIS and 178 healthy subjects. One mutation in ABCB4 (no. 69233, G>A) and two other mutations in ABCB11, reference single nucleotide polymorphism (rs)118109635 and rs497692, were identified in association with PIS (P<0.001, P=0.04 and P=0.02, respectively). A synonymous mutation at no. 69233 G>A was detected in exon 26 of ABCB4 in 23 heterozygous patients with PIS. This mutation was not detected in healthy individuals or in the Single Nucleotide Polymorphism Database. No. 69233 G>A in ABCB4 was not associated with altered protein expression but with a reduced rate of PIS recurrence (P=0.01). The missense mutation rs118109635 was located on exon 21 of ABCB11 and was associated with the increased expression of ABCB11 protein (P=0.032) as well as altered bile salt export pump function. Another synonymous mutation, rs497692 in exon 24 was reported to decrease ABCB11 protein expression (P=0.001). In addition, the mutations of ABCB11 were associated with preoperative jaundice (P<0.001 and P=0.03, respectively). Consistently decreased levels of ABCB11 protein were associated with recurrent episodes of cholangitis (P=0.006) and preoperative jaundice (P=0.015). By contrast, ABCB4 expression was not found to be associated with clinical manifestations of PIS.