Genome-wide copy number variant analysis reveals candidate genes associated with milk production traits in water buffalo (Bubalus bubalis).

Buffaloes are vital contributors to the global dairy industry. Understanding the genetic basis of milk production traits in buffalo populations is essential for breeding programs and improving productivity. In this study, we conducted whole-genome resequencing on 387 buffalo genomes from 29 diverse Asian breeds, including 132 river buffaloes, 129 swamp buffaloes, and 126 crossbred buffaloes. We identified 36,548 copy number variants (CNV) spanning 133.29 Mb of the buffalo genome, resulting in 2,100 CNV regions (CNVR), with 1,993 shared CNVR being found within the studied buffalo types. Analyzing CNVR highlighted distinct genetic differentiation between river and swamp buffalo subspecies, verified by evolutionary tree and principal component analyses. Admixture analysis grouped buffaloes into river and swamp categories, with crossbred buffaloes displaying mixed ancestry. To identify candidate genes associated with milk production traits, we employed 3 approaches. First, we used Vst-based population differentiation, revealing 11 genes within CNVR that exhibited significant divergence between different buffalo breeds, including genes linked to milk production traits. Second, expression quantitative loci analysis revealed differentially expressed CNVR-derived genes (DECG) associated with milk production traits. Notably, known milk production-related genes were among these DECG, validating their relevance. Last, a GWAS identified 3 CNVR significantly linked to peak milk yield. Our study provides comprehensive genomic insights into buffalo populations and identifies candidate genes associated with milk production traits. These findings facilitate genetic breeding programs aimed at increasing milk yield and improving quality in this economically important livestock species.

CX43 and oxidative stress are the targets of BCB staining to predict the developmental potential of buffalo oocytes.

This study used the brilliant cresyl blue (BCB) staining method to group buffalo oocytes (BCB+ and BCB-) and perform in vitro maturation, in vitro fertilization and embryo culture. At the same time, molecular biology techniques were used to detect gap junction protein expression and oxidative stress-related indicators to explore the molecular mechanism of BCB staining to predict oocyte developmental potential. The techniques of buffalo oocytes to analyse their developmental potential and used immunofluorescence staining to detect the expression level of CX43 protein, DCFH-DA probe staining to detect ROS levels and qPCR to detect the expression levels of the antioxidant-related genes SOD2 and GPX1. Our results showed that the in vitro maturation rate, embryo cleavage rate and blastocyst rate of buffalo oocytes in the BCB+ group were significantly higher than those in the BCB- group and the control group (p < .05). The expression level of CX43 protein in the BCB+ group was higher than that in the BCB- group both before and after maturation (p < .05). The intensity of ROS in the BCB+ group was significantly lower than that in the BCB- group (p < .05), and the expression levels of the antioxidant-related genes SOD2 and GPX1 in the BCB+ group were significantly higher than those in the BCB- group (p < .05). Brilliant cresyl blue staining could effectively predict the developmental potential of buffalo oocytes. The results of BCB staining were positively correlated with the expression of gap junction protein and antioxidant-related genes and negatively correlated with the reactive oxygen species level, suggesting that the mechanism of BCB staining in predicting the developmental potential of buffalo oocytes might be closely related to antioxidant activity.

Transcriptome-Wide Association Study Reveals Potentially Candidate Genes Responsible for Milk Production Traits in Buffalo.

Identifying key causal genes is critical for unraveling the genetic basis of complex economic traits, yet it remains a formidable challenge. The advent of large-scale sequencing data and computational algorithms, such as transcriptome-wide association studies (TWASs), offers a promising avenue for identifying potential causal genes. In this study, we harnessed the power of TWAS to identify genes potentially responsible for milk production traits, including daily milk yield (MY), fat percentage (FP), and protein percentage (PP), within a cohort of 100 buffaloes. Our approach began by generating the genotype and expression profiles for these 100 buffaloes through whole-genome resequencing and RNA sequencing, respectively. Through comprehensive genome-wide association studies (GWAS), we pinpointed a total of seven and four single nucleotide polymorphisms (SNPs) significantly associated with MY and FP traits, respectively. By using TWAS, we identified 55, 71, and 101 genes as significant signals for MY, FP, and PP traits, respectively. To delve deeper, we conducted protein-protein interaction (PPI) analysis, revealing the categorization of these genes into distinct PPI networks. Interestingly, several TWAS-identified genes within the PPI network played a vital role in milk performance. These findings open new avenues for identifying potentially causal genes underlying important traits, thereby offering invaluable insights for genomics and breeding in buffalo populations.

Novel Insight into the Potential Role of Acylglycerophosphate Acyltransferases Family Members on Triacylglycerols Synthesis in Buffalo.

Acylglycerophosphate acyltransferases (AGPATs) are the rate-limiting enzymes for the de novo pathway of triacylglycerols (TAG) synthesis. Although AGPATs have been extensively explored by evolution, expression and functional studies, little is known on functional characterization of how many members of the AGPAT family are involved in TAG synthesis and their impact on the cell proliferation and apoptosis. Here, 13 AGPAT genes in buffalo were identified, of which 12 AGPAT gene pairs were orthologous between buffalo and cattle. Comparative transcriptomic analysis and real-time quantitative reverse transcription PCR (qRT-PCR) further showed that both AGPAT1 and AGPAT6 were highly expressed in milk samples of buffalo and cattle during lactation. Knockdown of AGPAT1 or AGPAT6 significantly decreased the TAG content of buffalo mammary epithelial cells (BuMECs) and bovine mammary epithelial cells (BoMECs) by regulating lipogenic gene expression (p < 0.05). Knockdown of AGPAT1 or AGPAT6 inhibited proliferation and apoptosis of BuMECs through the expression of marker genes associated with the proliferation and apoptosis (p < 0.05). Our data confirmed that both AGPAT1 and AGPAT6 could regulate TAG synthesis and growth of mammary epithelial cells in buffalo. These findings will have important implications for understanding the role of the AGPAT gene in buffalo milk performance.

Subtotal colectomy, extended right hemicolectomy, left hemicolectomy, or splenic flexure colectomy for splenic flexure tumors: a network meta-analysis.

AIM: To perform a network meta-analysis of the current literature to evaluate the short-term and long-term outcomes of four operations for splenic flexure tumors. METHODS: An electronic literature search of PubMed, Baidu Scholar, EMBASE, and Cochrane Central Register of Controlled Trials databases was performed up to August 2020. A Bayesian network meta-analysis was utilized to compare the outcomes involved in subtotal colectomy (STC), extended right hemicolectomy (ERHC), standard left hemicolectomy (LHC), and splenic flexure colectomy (SFC) by using R software. RESULTS: A total of 10 non-randomized studies were included in this meta-analysis. There was no statistically significant difference among these 4 surgical techniques in terms of the utilization rate of minimally invasive surgery, reoperative surgery, anastomotic dehiscence, mortality, the proportion of patients with the number of lymph nodes harvested ≥ 12, local recurrence, distant recurrence and overall survival. Although ERHC was associated with a higher risk of postoperative ileus (ERHC vs SFC, OR = 6.4, 95% CI 1.4-45.0, P = 0.019), it has an advantage of a higher rate of primary anastomosis (ERHC vs LHC, OR = 4.2, 95% CI 1.3-18.0, P = 0.019) and a non-significant trend for lower anastomotic dehiscence when compared with more restrict resections. CONCLUSION: SFC, LHC, ERHC and STC for the curative resection of splenic flexure tumors provide similar survival. An individualized surgical plan considering both long-term and short-term outcomes is necessary to select the appropriate operations.

Timing to achieve the best recurrence-free survival after neoadjuvant chemoradiotherapy in locally advanced rectal cancer: experience in a large-volume center in China.

AIM: To identify the optimal interval from the end of neoadjuvant chemoradiotherapy to surgery (CRT-surgery interval) based on long-term oncological outcome of locally advanced rectal cancer (LARC). METHODS: Retrospective data analysis is reported from patients diagnosed with cT3 or T4 or TxN+ rectal cancer who underwent neoadjuvant treatment and curative-intent surgery between January 2010 and December 2018. With a priority focus on the effect of interval on oncological prognosis, we used recurrence-free survival (RFS) as the primary endpoint to determine the best cutoff point of time intervals. Then, the short-term and long-term outcomes of patients from longer and shorter interval groups were compared. RESULTS: Data from 910 patients were analyzed, with 185 patients who achieved pCR (20.3%). The trend for increased rates of pCR for groups with a prolonged time interval was not observed (P = 0.808). X-tile determined a cutoff value of 10.5 weeks, and the population was divided into longer (> 10 weeks) and shorter (≤ 10 weeks) interval groups. The shorter interval was associated with a higher wound infection rate (4.7% vs. 1.1%, P = 0.031), but other postoperative complications did not differ between the groups. The 5-year RFS rate was significantly higher in patients in a longer group than those in the shorter weeks group (86.8% vs. 77.8%, P = 0.016). The 5-year OS rates between groups were similar (84.1% vs. 82.5%, P = 0.257). Local recurrence and lung metastases rates were higher in shorter interval group than those of longer group (local recurrence rate: 1.7% vs. 5.1%, P = 0.049; lung metastases rate: 5.7% vs. 10.7%, P = 0.047). Cox multivariate regression analysis confirmed the CRT-surgery interval (HR = 0.599, P = 0.045) to be an independent prognostic factor of RFS. CONCLUSION: This study is the first, to the best of our knowledge, to define the optimal CRT-surgery interval based on RFS as the primary endpoint. Prolonging the waiting period to 10 weeks after the completion of CRT with additional chemotherapy cycles during the interval period might be a promising option to improve oncological survival in LARC patients treated with CRT and TME without compromising the surgical safety. Further randomized controlled trials investigating this are warranted to prove a clearly causality.

Prognostic significance of carcinoembryonic antigen combined with carbohydrate antigen 19-9 following neoadjuvant chemoradiotherapy in patients with locally advanced rectal cancer.

AIM: The clinical significance of carcinoembryonic antigen (CEA) combined with carbohydrate antigen 19-9 (CA19-9) in patients with rectal cancer is not well established. The aim of this study was to determine the prognostic value of these combined tumour markers in patients with locally advanced rectal cancer (LARC) treated with neoadjuvant chemoradiotherapy (nCRT). METHOD: A total of 687 consecutive patients with LARC who underwent nCRT and radical surgery were analysed. Tumour characteristics, recurrence-free survival (RFS) and overall survival (OS) were compared according to the number of elevated tumour markers measured before and after nCRT. In addition, the prognostic significance of perioperative changes in the combined tumour markers was further evaluated. RESULT: The RFS and OS rates decreased in a stepwise manner in association with the number of elevated pre- and post-nCRT tumour markers (all p < 0.05). Multivariate analysis showed that only the number of elevated post-nCRT tumour markers was an independent prognostic factor (both p < 0.05). For 311 patients with elevated pre-nCRT tumour markers, normalization of the tumour markers after nCRT was an independent prognostic protective factor (both p < 0.05), and patients with both markers elevated post-nCRT had a 2.5- and 3.7-fold increased risk of recurrence and death, respectively (p < 0.05). Furthermore, normalization of post-nCRT tumour markers after surgery was also closely related to an improved prognosis. CONCLUSION: This combination of post-nCRT tumour markers can accurately predict the long-term survival of patients with LARC treated with nCRT and curative resection, and normalization of the combined tumour markers after either nCRT or surgery was associated with better survival.

Overexpressed CES2 has prognostic value in CRC and knockdown CES2 reverses L-OHP-resistance in CRC cells by inhibition of the PI3K signaling pathway.

CES-2 (carboxylesterase-2) belongs to the carboxylesterase gene family, which plays crucial roles in lipid mobilization and chemosensitivity to irinotecan. However, its role in chemosensitivity to oxaliplatin (L-OHP) remains unclear. Herein, L-OHP-resistant cells (HCT-116L and RKOL) were established by increasing the concentration of L-OHP. The results showed that CES2 expression was upregulated in L-OHP-resistant tissues and cells lines (both P < 0.01). Low expression of CES2 correlated with a better survival, and the results were further confirmed in the R2 platform: a biologist friendly web-based genomics analysis and visualization application. Downregulation of CES2 suppressed cell proliferation, induced apoptosis and reversed L-OHP resistance by medicating the PI3K signaling pathway in L-OHP-resistant cells. However, both PI3K inhibitor (LY294002) and activator (IGF-1) could not medicate CES2 expression. These findings indicated that CES2 may be utilized as a novel biomarker and therapeutic target for L-OHP resistance in CRC treatment.

Incidence of and Risk Factors for Gastroepiploic Lymph Node Involvement in Patients with Cancer of the Transverse Colon Including the Hepatic Flexure.

BACKGROUND: To define the incidence of gastroepiploic lymph node (GLN) metastasis in patients with cancer of the transverse colon, including the hepatic flexure, and to identify the preoperative predictors of GLN involvement in a large-volume center in China. METHODS: This retrospective monocentric cross-sectional study respected the STROBE statement. Of 3208 consecutive patients who underwent colon cancer resection, a total of 371 patients with cancer of the transverse colon including the hepatic flexure who underwent complete mesocolic excision and GLN resection in our center were retrospectively reviewed between November 2010 and November 2017. Logistic regression was performed to identify predictors of GLN metastasis. Endoscopic obstruction was defined as a luminal obstruction of the colon severe enough to prevent the colonoscope from passing beyond the tumor regardless of the presenting symptoms. RESULTS: The GLN involvement rate was 4.0 (2.0-6.1)%. Patients who had GLN involvement had a significantly higher rate of endoscopic obstruction (P = 0.030), higher rate of signet ring adenocarcinoma or lymphovascular invasion (P < 0.05), higher preoperative CEA level (P = 0.037), more advanced pN stage (P < 0.001) and more advanced M stage (P = 0.003) than the patients without GLN involvement. ROC curve analyses showed that the cutoff value for CEA was 17.0 ng/ml (46.7% sensitivity, 84.3% specificity, P = 0.037) for the prediction of GLN metastasis. Multivariate analysis showed that endoscopic obstruction, signet ring adenocarcinoma, a CEA level ≥17 ng/ml and M1 stage were independently correlated with the GLN metastasis. CONCLUSION: The incidence rate of GLN metastasis was low. To the best of our knowledge, the present study was the first to evaluate the preoperative predictors of GLN metastasis. Combinations of predictive factors may be useful for stratifying patients at high risk of GLN metastasis.

Pelvimetric and Nutritional Factors Predicting Surgical Difficulty in Laparoscopic Resection for Rectal Cancer Following Preoperative Chemoradiotherapy.

AIM: Laparoscopic total mesorectal excision (LaTME) following preoperative chemoradiotherapy (PCRT) in locally advanced rectal cancer (LARC) is technically demanding. The present study is intended to evaluate predictive factors of surgical difficulty of LaTME following PCRT by using pelvimetric and nutritional factors. METHOD: Consecutive LARC patients receiving LaTME after PCRT were included. Surgical difficulty was classified based upon intraoperative (operation time, blood loss, and conversion) and postoperative outcomes (postoperative hospital stay and morbidities). Pelvimetry was performed using preoperative T2-weighted MRI. Nutritional factors such as albumin-to-globulin ratio (AGR) and prognostic nutritional index (PNI) were calculated. Multivariable logistic analysis was used to identify predictors of high surgical difficulty. A predictive nomogram was developed and validated internally. RESULTS: Among 294 patients included, 36 (12.4%) patients were graded as high surgical difficulty. Logistic regression analysis demonstrated that previous abdominal surgery (OR = 6.080, P = 0.001), tumor diameter (OR = 1.732, P = 0.003), intersphincteric resection (vs. low anterior resection, OR = 13.241, P < 0.001), interspinous distance (OR = 0.505, P = 0.009), and preoperative AGR (OR = 0.041, P = 0.024) were independently predictive of high surgical difficulty of LaTME after PCRT. Then, a predictive nomogram was built (C-index = 0.867). CONCLUSION: Besides previous abdominal surgery, type of surgery (intersphincteric resection), tumor diameter, and interspinous distance, we found that preoperative AGR could be useful for the prediction of surgical difficulty of LaTME after PCRT. A predictive nomogram for surgical difficulty may aid in planning an appropriate approach for rectal cancer surgery after PCRT.

Combination of Preoperative Plasma Fibrinogen and Neutrophil-to-Lymphocyte Ratio (the F-NLR Score) as a Prognostic Marker of Locally Advanced Rectal Cancer Following Preoperative Chemoradiotherapy.

BACKGROUND: Coagulation and inflammation play important roles in tumor progression. This study aimed to explore the prognostic impact of combined analysis of fibrinogen and neutrophil-to-lymphocyte (NLR) ratio (F-NLR score) in locally advanced rectal cancer (LARC) receiving preoperative chemoradiotherapy (pCRT) and radical surgery. METHOD: Totally 317 patients were included. X-tile analysis was used to determine the optimal cutoff values of preoperative fibrinogen and NLR. F-NLR scores were defined as 2 (both high fibrinogen and NLR), 1 (one of these abnormalities), or 0 (neither abnormality). Time-dependent ROC analysis was used to evaluate the predictive accuracy of fibrinogen, NLR, and F-NLR score. Cox regression analysis was performed to evaluate the prognostic impact of the F-NLR score. A predictive nomogram for disease-free survival (DFS) was developed and validated internally. RESULTS: One hundred and seventeen (36.9%), 156 (49.2%), and 44 (13.9%) patients had F-NLR score of 0, 1, and 2, respectively. Higher F-NLR score was associated with poorly differentiated tumors, deeper tumor invasion, lymph node metastasis, and more advanced pTNM stage (all P < 0.05). The 5-year OS rates in the F-NLR 0, 1, and 2 groups were 93.6%, 87.3%, and 68.4%, respectively (P < 0.001), while the 5-year DFS rates were 91.8%, 76.8%, and 56.1%, respectively (P < 0.001). Cox regression analysis demonstrated that F-NLR score (F-NLR 1, HR = 2.021, P = 0.046; F-NLR 2, HR = 3.356, P = 0.002), pTNM stage III (HR = 3.109, P = 0.009), and circumferential resection margin (CRM) involvement (HR = 3.120, P = 0.021) were independently associated with DFS. A nomogram for DFS was developed (C-index 0.708). CONCLUSION: F-NLR score is a promising predictor for disease recurrence in LARC patients after pCRT.

Is Early Initiation of Adjuvant Chemotherapy Beneficial for Locally Advanced Rectal Cancer Following Neoadjuvant Chemoradiotherapy and Radical Surgery?

AIM: This study aimed to evaluate whether earlier initiation (< 4 weeks) of adjuvant chemotherapy (ACT) confer any oncological benefits for locally advanced rectal cancer (LARC) patients undergoing neoadjuvant chemoradiotherapy (nCRT) and radical surgery. METHOD: Clinicopathological and survival outcomes were compared. Propensity score matching (PSM) was performed to adjust for differences between groups. Cox regression analysis was performed to evaluate the impact of earlier ACT initiation on overall survival (OS) and disease-free survival (DFS). RESULTS: Totally, 443 eligible patients were included. More laparoscopic surgeries, less postoperative complications, and more ACT completion were observed in patients whose ACT was initiated within 4 weeks after surgery (all P < 0.001). With a mean follow-up of 59 months, the 5-year OS and DFS rate was 89.8% and 82.0% in the early group, significantly higher than 81.6% and 73.1% in the late group (P = 0.007, and P = 0.022, respectively). After PSM, the 5-year OS and DFS rate was 90.9% and 84.4% in the early group, significantly higher than 83.4% and 68.8% in the late group (P = 0.047, and P = 0.017, respectively). Cox regression analysis demonstrated that time to ACT initiation (early vs. late, HR = 0.486, P = 0.008) was independently associated with OS. CONCLUSION: Early initiation of ACT (<4 weeks) confers a survival benefit, and is an independent prognostic factor of OS in LARC patients following nCRT. Further investigations are needed to define the role of earlier initiation of ACT in patients with LARC after nCRT.

A nomogram for predicting rectovaginal fistula after low anterior resection for rectal cancer.

PURPOSE: Rectovaginal fistula (RVF) is a complicated and troublesome complication of low anterior resection (LAR) for rectal cancer. We aimed to investigate the risk factors for post-LAR RVF and develop a predictive nomogram. METHODS: We performed a retrospective analysis of 821 female patients with rectal cancer who underwent LAR between October 2010 and October 2018. Logistic regression was performed to identify risk factors. A nomogram was developed to predict RVF. RESULTS: The incidence of post-LAR RVF was 3.4% (28/821). A multivariate analysis showed that the preoperative serum hemoglobin level (OR 2.449, 95% CI 1.144-5.239), the distance between the tumor and anal verge (OR 4.158, 95% CI 1.392-12.418), surgical procedure (OR 2.369, 95% CI 1.117-5.027), hysterectomy (OR 2.996, 95% CI 1.106-8.833), and bilateral oophorectomy (OR 5.823, 95% CI 1.639-20.689) were significantly associated with the development of RVF. A nomogram was developed, which showed a C-index of 0.824 (95% CI 0.730-0.918) and an adjusted C-index of 0.790. CONCLUSION: This study identified the preoperative serum hemoglobin level, the distance between the tumor and the anal verge, the type of surgical procedure, hysterectomy, and bilateral oophorectomy as predictors of post-LAR RVF. A nomogram was successfully developed. It could aid in the prediction of RVF in patients undergoing LAR.

Hypermethylated and downregulated MEIS2 are involved in stemness properties and oxaliplatin-based chemotherapy resistance of colorectal cancer.

The resistance against oxaliplatin (L-OHP) based regimens remains a major obstacle for its efficient usage in treating metastatic colorectal cancer (mCRC). In this study, we performed weighted gene coexpression network analysis (WGCNA) to systematically screen the relevant hub genes for L-OHP resistance using the raw microarray data of 30 consecutive mCRC samples from our earlier study (GSE69657). The results were further confirmed through datasets from Gene Expression Omnibus (GEO). From L-OHP resistance module, nine genes in both the coexpression and protein-protein interaction networks were chosen as hub genes. Among these genes, Meis Homeobox 2 (MEIS2) had the highest correlation with L-OHP resistance (r = -0.443) and was deregulated in L-OHP resistant tissues compared with L-OHP sensitive tissues in both our own dataset and GSE104645 testing dataset. The receiver operating characteristic curve validated that MEIS2 had a good ability in predicting L-OHP response in both our own dataset (area under the curve [AUC] = 0.802) and GSE104645 dataset (AUC = 0.746). Then, the down expression of MEIS2 was observed in CRC tissue compared with normal tissue in 12 GEO-sourced datasets and The Cancer Genome Atlas (TCGA) and was correlated with poor event-free survival. Furthermore, analyzing methylation data from TCGA showed that MEIS2 had increased promoter hypermethylation. In addition, MEIS2 expression was significantly decreased in CRC stem cells compared with nonstem cells in two GEO datasets (GSE14773 and GSE24747). Further methylation analysis from GSE104271 demonstrated that CRC stem cells had higher MEIS2 promoter methylation levels in cg00366722 and cg00610348 sites. Gene set enrichment analysis showed that MEIS2 might be involved in the Wnt/ß-catenin pathway. In the overall view, MEIS2 had increased promoter hypermethylation and was downregulated in poor L-OHP response mCRC tissues. MEIS2 might be involved in the Wnt/ß-catenin pathway to maintain CRC stemness, which leads to L-OHP resistance.

Coexpression network analysis linked H2AFJ to chemoradiation resistance in colorectal cancer.

Neoadjuvant chemoradiotherapy (CRT) resistance is a complex phenomenon and it remains a major problem for patients with a priori resistant tumor. Therefore, there is a strong need to investigate molecular biomarkers which may guide for treatment decision-making. In our study, weighted gene coexpression network analysis was applied to identify CRT-resistance hub modules in 12 colorectal cancer (CRC) cell lines with different CRT sensitivities from GSE20298 data set. The green module and purple module had the highest correlations with CRT resistance. Gene ontology enrichment analysis indicated that the function of these two modules focused on interferon-mediated signaling pathway, immune response, chromatin modulation, Rho GTPases activities, and regulation of apoptotic process. Then, 15 hub genes in both the coexpression and protein-protein interaction networks were selected. Among these hub genes, higher H2A histone family member J (H2AFJ) expression was independently validated in patient cohorts from two testing data sets of GSE46862 and GSE68204 to be related to CRT resistance. The receiver operating characteristic curve showed that H2AFJ could efficiently distinguish CRT-resistance cases from CRT-sensitive cases in another two testing data sets. Furthermore, meta-analysis of 12 Gene Expression Omnibus-sourced data sets showed that H2AFJ messenger RNA levels were significantly higher in CRC tissues than in normal colon tissues. High H2AFJ expression was correlated with a significant worse event- and relapse-free survival by analyzing the data from the R2: Genomics Analysis and Visualization Platform. Gene set enrichment analysis determined that the mechanism of H2AFJ-mediated CRT resistance might involve the ERK5 (MAPK7), human immunodeficiency virus Nef (HIV Nef), and inflammatory pathways. This study is the first, to the best of our knowledge, to implicate and verify H2AFJ as an effective new marker for CRT response prediction.

Downregulated SPINK4 is associated with poor survival in colorectal cancer.

BACKGROUND: SPINK4 is known as a gastrointestinal peptide in the gastrointestinal tract and is abundantly expressed in human goblet cells. The clinical significance of SPINK4 in colorectal cancer (CRC) is largely unknown. METHODS: We retrieved the expression data of 1168 CRC patients from 3 Gene Expression Omnibus (GEO) datasets (GSE24551, GSE39582, GSE32323) and The Cancer Genome Atlas (TCGA) to compare the expression level of SPINK4 between CRC tissues and normal colorectal tissues and to evaluate its value in predicting the survival of CRC patients. At the protein level, these results were further confirmed by data mining in the Human Protein Atlas and by immunohistochemical staining of samples from 81 CRC cases in our own center. RESULTS: SPINK4 expression was downregulated in CRC compared with that in normal tissues, and decreased SPINK4 expression at both the mRNA and protein levels was associated with poor prognosis in CRC patients from all 3 GEO datasets, the TCGA database and our cohort. Additionally, lower SPINK4 expression was significantly related to higher TNM stage. Moreover, in multivariate regression, SPINK4 was confirmed as an independent indicator of poor survival in CRC patients in all databases and in our own cohort. CONCLUSIONS: We concluded that reduced expression of SPINK4 relates to poor survival in CRC, functioning as a novel indicator.

Prognostic significance of neoadjuvant rectal score in locally advanced rectal cancer after neoadjuvant chemoradiotherapy and construction of a prediction model.

AIM: To evaluate the prognostic significance of neoadjuvant rectal (NAR) score after neoadjuvant chemoradiotherapy (nCRT) for locally advanced rectal cancer (LARC), and to develop a nomogram predicting disease-free survival (DFS). METHOD: A total of 522 LARC patients undergoing nCRT and surgery were included. NAR scores were calculated using the equation [5pN-3(cT-pT) + 12]^2/9.61, and classified as low (<8), intermediate (8-16), and high (>16). Clinicopathological and survival outcomes were compared. Cox regression analysis was performed to identify risk factors of DFS. A predicting nomogram was developed and validated internally. RESULTS: For NAR score classification, 193 (37.0%) were low, 183 (35.0%) were intermediate, and 146 (28.0%) were high. Higher NAR score was associated with fewer pCR, lower tumor regression grade (TRG), and higher ypTNM stage. A total of 5-year DFS for low, intermediate, and high NAR groups was 85.6%, 71.9%, and 47.2%, respectively (P < 0.001). NAR score (HR = 2.488, P = 0.002), TRG (HR = 2.811, P = 0.047), CRM involvement (HR = 2.703, P = 0.002), and IMA nodal metastasis (HR = 2.441, P = 0.001) were independent prognostic factors of DFS. A predicting nomogram was developed with C-index of 0.701. CONCLUSION: NAR score could help in predicting DFS after nCRT. A nomogram was developed to identify subpopulations with aggressive tumors during clinical decision-making.

Risk Factors for Early Postoperative Small Bowel Obstruction after Elective Colon Cancer Surgery: An Observational Study of 1,244 Consecutive Patients.

BACKGROUND: Early postoperative small bowel obstruction (EPSBO) is a common complication following colon cancer surgery. EPSBO is associated with increased hospital stays, mortality rates, and healthcare costs. The purpose of this study was to identify risk factors for EPSBO following elective colon cancer surgery. STUDY DESIGN: We retrospectively reviewed the clinicopathological variables of 1,244 patients with colon cancer who underwent partial colectomy from January 2000 to December 2014. A multivariable logistic regression model was used to identify risk factors for EPSBO. RESULTS: The EPSBO rate was 3.5%. In multivariate analysis, preoperative bowel obstruction (OR 2.378; 95% CI 0.986-5.735, p = 0.054), weight loss >10% of body weight (OR 3.029; 95% CI 1.000-9.178, p = 0.05), albumin level (in g/L; OR 0.966; 95% CI 0.937-0.996, p = 0.024), and surgical duration (in min; OR 1.008; 95% CI 1.003-1.012, p = 0.003) were significant predictors of EPSBO. CONCLUSION: EPSBO is more likely to develop in the presence of poor systemic conditions (e.g., weight loss >10% of body weight, hypoalbuminemia, and preoperative bowel obstruction) and following operations of longer duration. These predictors may facilitate the stratification of patients at risk for EPSBO following surgery for elective colon cancer.

Molecular characterisation of the buffalo SCAP gene and its association with milk production traits in water buffaloes.

The study reported in this Research Communication was conducted to investigate the molecular characterisation of buffalo SCAP gene, expression analysis, and the association between single nucleotide polymorphisms and milk production traits in 384 buffaloes. Sequence analysis revealed the SCAP gene had an open reading frame of 3837 bp encoding 1279 amino acids. A ubiquitous expression profile of SCAP gene was detected in various tissues with extreme predominance in the mammary gland during early lactation. Moreover, eleven SNPs in buffalo SCAP gene were identified, six of them (g.1717600A>G, g.1757922C>T, g.1758953G>A, g.1759142C>T, g.1760740G>A, and g.1766036T>C) were found to be significantly associated with 305-day milk yield. Thus, buffalo SCAP could sever as a candidate gene affecting milk production traits in buffalo and the identified SNPs might potentially be genetic markers.

A nomogram to predict distant metastasis after neoadjuvant chemoradiotherapy and radical surgery in patients with locally advanced rectal cancer.

PURPOSE: To compare distant metastasis (DM) in locally advanced rectal cancer (LARC) patients treated with neoadjuvant chemoradiotherapy (nCRT) and surgery alone, and to develop a predictive nomogram for DM following nCRT. METHODS: Propensity-scoring match analysis was performed to compare DM in LARC treated with nCRT (n = 375) and surgery alone (n = 375). Cox regression was performed to identify predictors of DM following nCRT. A nomogram was developed and validated by internal (n = 425) and external validation (n = 97). RESULTS: The 5-year local recurrence rate was significantly lower in the nCRT group (5.6% vs. 10.4%; P = 0.020). The 5-year DM rates (nCRT vs. surgery alone: 25.3% vs. 24.4%; P = 0.235) were similar between groups. Cox regression showed that the post-nCRT pathologic stage (ypTNM stage, OR = 2.022, P = 0.002), IMA nodal metastasis (OR = 2.171, P = 0.023), and CRM involvement (OR = 2.535, P = 0.016) were independently associated with DM following nCRT. A predictive nomogram was developed with a C-index of 0.70 on internal validation, and 0.71 on the external validation. CONCLUSION: NCRT improved local control, but not distant metastasis. A nomogram to predict 3- and 5-year DM rates, using clinicopathological parameters, was successfully developed. This prognostic tool could support decision-making in clinical practice and follow-up strategies.