[Study on the application value of fecal SDC2 gene methylation detection in colorectal cancer screening of urban residents in Zengcheng District in Guangzhou City].

Objective: To investigate the application value of fecal Syndecan-2 (SDC2) gene methylated SDC2 (mSDC2) detection in colorectal cancer (CRC) screening among urban residents in Guangzhou City. Methods: A cross-sectional study was conducted in Shitan Town, Zengcheng District, Guangzhou City from July to December 2022. A community-based screening program for CRC was conducted among residents aged 40-74 years old. mSDC2 detection was employed in the participants, and those with positive results should be recommended to receive colonoscopy examination. The positive rate of mSDC2 detection, colonoscopy compliance rate, detection rate of intestinal lesions and clinicopathological characteristics were observed. The relationship between cycle threshold (CT) value of mSDC2 and intestinal lesions was explored. Further, the cost-effectiveness of screening was evaluated. Results: A total of 8 189 fecal samples were collected from 8 877 participants with the recovery rate of 92.25%. 8 048 qualified samples were enrolled in this study, consisted of 3 182 males (39.54%) and 4 866 females (60.46%), with the average age of 56 years old (40-74 years). The positive rate of mSDC2 detection was 7.99% (643/8 048), and the compliance rate of colonoscopy was 73.10% (470/643). 20 cases (4.25%) of colorectal cancer, 109 cases (23.19%) of advanced adenoma, 145 cases (30.85%) of non-advanced adenoma, 79 cases (16.81%) of polyps were detected. The detection rate of intestinal lesions was 75.11% and indicated significant differences in gender and age. 20 CRCs included 15 of stage 0-I, 4 of stage Ⅱ-Ⅲ and 1 of unknown stage. The CT value of mSDC2 was negatively correlated with the proportion of advanced colorectal neoplasms (χ2=16.063, P<0.001). The total cost of the screening was 4.339 5 million yuan, the screening benefit was 28.506 2 million yuan, and the benefit-cost ratio was 6.57. Conclusion: The CRC screening strategy of fecal mSDC2 detection combined with colonoscopy has high colonoscopy compliance and detection rate of intestinal lesions, which is conducive to the detection of early CRCs, and has good cost-effectiveness. This study suggests that this method may be applied to the general CRC screening in China and contribute to the prevention of CRC. The CT value of mSDC2 may have a certain suggestion on the malignant degree of intestinal tumors.

[Risk factors for delayed gastric emptying after laparoscopic pancreaticoduodenectomy: a single-center experience of 1 000 cases].

Objective: To explore the causes and summarize the treatment experience for clinically relevant delayed gastric emptying(DGE) after laparoscopic pancreaticoduodenectomy(LPD). Methods: The clinical data of 1 000 patients who underwent LPD in the Department of Liver Transplantation and Hepatobiliary Surgery,Shandong Provincial Hospital Affiliated to Shandong First Medical University between March 2017 and September 2022 was retrospectively collected. There were 640 males and 360 females,with an age of (60.1±11.4)years(range: 13 to 93 years),and 590 patients were older than 60 years. Depending on the severity of DGE,patients were divided into a clinically relevant DGE group and a 0/A grade DGE group. The comparison between the two groups was performed by the χ2 test,Fisher's exact probability method,t test or the rank sum test,and the effects of various treatment strategies for clinically relevant DGE were evaluated. Results: LPD was conducted successfully in all 1 000 patients,with a surgical time of (344.8±103.6)minutes(range:160 to 450 minutes) and intraoperative blood loss (M(IQR)) of 100 (150) ml(range:50 to 1 000 ml). A total of 74 patients(7.4%) developed clinically relevant DGE. Compared to those in the 0/A grade DGE group,patients in the clinically relevant DGE group had a higher preoperative body mass index of ((24.9±3.5)kg/m2 vs. (23.9±3.3)kg/m2,t=-2.419,P=0.016),more postoperative bile leakage(51.4%(38/74) vs. 10.8%(100/926)),pancreatic fistula(59.5%(44/74) vs. 22.9%(212/926)),abdominal infection(74.3%(55/74) vs.14.6%(135/926)),and abdominal bleeding(43.2%(32/74) vs. 11.3%(105/926))(all P<0.05). Among these patients,10 cases(13.5%) received enteral nutrition treatment,22 cases(29.7%) received parenteral nutrition treatment,and 42 cases(56.8%) received a combination of enteral and parenteral nutrition treatment. The time for patients to return to a normal diet was 21(14)days (range: 8 to 85 days). Compared to those who received only enteral(23.5(27.0)days) or parenteral nutrition treatment(15.5(11.0)days),patients who received a combination of enteral and parenteral nutrition treatment(25.5(31.0)days) had a longer time to return to a normal diet (Z=20.019,P<0.01). Among the 60 patients who developed secondary DGE,48 cases(80.0%) received ultrasound-guided puncture and drainage treatment,while 12 cases(20.0%) only received anti-infection treatment. The patients in the non-puncture drainage group had a longer time to return to a normal diet than those in the puncture drainage group (26.5(12.5)days vs. 20.0(11.0)days, Z=-2.369,P=0.018). Conclusions: Patients with clinically relevant DGE after LPD had a higher proportion of postoperative complications such as pancreatic fistula,biliary fistula and abdominal infection. A combination of enteral and parenteral nutrition treatment is needed for patients with a long-term course of DGE."Smooth" drainage and ani-infectious therapy could contribute to the recovery of DGE.

[Scientific and rigorous data registration of patients with gastric cancer is the cornerstone of high-quality surgical research on gastric cancer: lessons learned from the Dutch upper gastrointestinal cancer audit (DUCA)].

The postoperative 30-day mortality and morbidity of gastric cancer surgery has markedly improved over the past years due to minimally invasive techniques, perioperative rehabilitation programs and centralization of care. However, there is still need for improvement as postoperative complications may have a serious negative impact on the efficacy of surgical treatment of gastric cancer. High-quality clinical research is a very important tool to analyze treatment outcomes and evaluate new treatment strategies. The meticulous registration of gastric cancer patient data is the basis of high-quality surgical research. In the past 11 years, the Dutch upper gastrointestinal cancer audit (DUCA) database has vast experience in data registration and maintenance of patients with upper gastrointestinal cancer. The effective measures it has taken in data registration, data quality control, data application and use, and data security have maintained quality at a high level. These data has been used for medical care quality monitoring and scientific research leading to a positive impact on the postoperative short-term outcomes of patients with upper gastrointestinal cancer. The work of DUCA may be a good incentive for the setup of population-based databases and clinical research in other countries.

[Safety and efficacy of a new domestic distal perforated stent graft in the treatment of Stanford type B aortic dissection].

Objective: To evaluate the safety and efficacy of a new domestic distal perforated stent graft (Talos stent) in the treatment of Stanford type B aortic dissection (TBAD). Methods: Twenty-five patients with TBAD treated with Talos stent in Yan'an Hospital Affiliated to Kunming Medical University from February 2018 to December 2019 were selected as the research subjects. Intraoperative angiography was performed to determine the number of branch arteries that remained after stent release. On postoperative day 5 (POD5), the pain intensity of the patients was evaluated by visual analog scale (VAS). The computed tomography angiography (CTA) of the patients before operation, 6 months and 12 months after operation were compared including aortic diameter, true lumen diameter, and false lumen diameter at the level of tracheal bifurcation. Follow-up was performed 1 month, 6 months, 12 months, and 24 months after surgery, and the occurrence of stent-related adverse events, reoperation and survival rate were recorded. Results: The enrolled patients included 19 males and 6 females, aged (52.6±11.1) years. Intraoperative angiography showed that 4 (1, 7) branch arteries were preserved, and the VAS score was 1 (0, 1) on POD5. The aortic diameters at the level of the tracheal bifurcation were (34.9±1.1) mm, (34.6±0.9) mm and (34.8±1.0) mm before surgery, 6 months and 12 months after surgery, and there was no significant difference (P=0.926); the diameters of the main true lumen at the level of the tracheal bifurcation were (13.3±1.6) mm, (21.8±1.0) mm and (22.3±1.1) mm before surgery, 6 months and 12 months postoperatively, while the diameters of the main false lumen at the level of the tracheal bifurcation were (20.8±2.2) mm, (4.5±1.5) mm, and (4.6±1.7) mm, respectively. Compared with before surgery, the diameter of true lumen increased significantly 6 months and 12 months after surgery (both P<0.001), while the diameter of false lumen decreased (both P<0.001). No stent-related adverse events occurred within 30 days after surgery, no secondary operations occurred within 12 months after surgery, no type Ⅰ and type Ⅲ endoleaks, no deaths or cases of paraplegia were reported, and the stent structure and position remained good. There were no deaths or paraplegia cases 24 months postoperatively, and no stent-related adverse events occurred. Conclusion: Using Talos stent in the treatment of TBAD can effectively help remodel the aorta, while preserve the intercostal artery and spinal artery, with good clinical effect and safety.

Identification of ferroptosis-related genes for the prediction of prognosis and chemotherapy benefit of gastric cancer.

OBJECTIVE: As a newly reported programmed cell death, ferroptosis plays a crucial role in tumor progression. We aimed to comprehensively analyze the prognostic value of ferroptosis-related genes and identify the genes for the prediction of prognosis and chemotherapy benefit of gastric cancer (GC) patients. MATERIALS AND METHODS: Public microarray data and corresponding clinical information of GC were downloaded from GEO databases. Patients in GSE66229 were randomly separated into discovery and internal validation at a ratio of 2:1. GSE15459 was set as the external validation set. LASSO Cox regression model was performed to identify the most significant prognostic ferroptosis-related genes. RESULTS: Twenty ferroptosis-related genes were finally identified to establish the predictive signature. In the discovery data set, the signature could divide patients into low- and high-risk groups with significantly different overall survival (HR: 2.11, 95% CI: 1.40-3.17, p<0.001). These results were successfully validated in the internal validation cohort (HR 2.04, 95% CI 1.18-3.52; p=0.008) and external validation cohort (HR 4.87, 95% CI 2.99-7.93; p=0.008). Survival ROC at 5-year revealed a remarkably higher predictive ability of the ferroptosis classifier (AUC=0.835) compared with other clinical features in predicting prognosis. Further GSEA analysis showed that samples of high risk were related to several established tumor invasion and metastatic signaling pathways. Further experiments revealed that VLDLR and GCH1 were two newly identified genes associated with chemotherapy sensitivity in GC. CONCLUSIONS: The developed ferroptosis gene-set based prognostic signature indicated superior prognostic and predictive value, suggesting new possibilities for individualized treatment of GC patients.

[Bowel preparation before elective surgery for colorectal cancer].

For elective surgery of colorectal cancer, current evidence supports preoperative mechanical bowel preparation combined with oral antibiotics. Meanwhile, for patients with varied degrees of intestinal stenosis, individualized protocol is required to avoid adverse events. We hereby summarize recent high-quality evidences and updates of guidelines and consensus, and recommend stratified bowel preparation based on the clinical practice of our institute as follows. (1) For patients with unimpaired oral intake, whose tumor can be passed by colonoscopy, mechanical bowel preparation and oral antibiotics are given. (2) For patients without symptoms of bowel obstruction but with impaired oral intake or incomplete colonoscopy due to tumor-related stenosis, small-dosage laxative is given for several days before surgery, and oral antibiotics the day before surgery. (3) For patients with bowel obstruction, mechanical bowel preparation or enema is not indicated. We proposed this evidence-based, individualized protocol for preoperative bowel preparation for the reference of our colleagues, in the hope of improving perioperative outcomes and reducing adverse events.

[The relationship between insulin resistance and risk of long-term mortality in people without diabetes: a 30-year follow-up of the Daqing Diabetes Study].

Objective: To determine whether insulin resistance is associated with all-cause mortality in subjects without diabetes. Methods: A total of 505 participants without diabetes, 198 with normal glucose tolerance (NGT) and 307 with impaired glucose tolerance (IGT), were recruited from the Daqing Diabetes Study. The participants were followed up for 30 years. They were stratified into three groups (tertiles) according to baseline homeostasis model assessment of insulin resistance(HOMA-IR) levels, as the HOMA-IR 0, the HOMA-IR 1 and the HOMA-IR 2 groups, to assess the predictive effect of insulin resistance on risk of all-cause mortality. Results: During the 30-year follow-up, 52, 56 and 78 participants died across the three HOMA-IR groups, respectively. The corresponding mortality per 1 000 person-years (95%CI) were 12.12 (9.56-15.01), 13.10 (10.46-16.03) and 19.91 (16.73-23.15), respectively. Participants in the HOMA-IR 2 group had a significantly higher risk of death than those in the HOMA-IR 0 group after adjustment of age, sex and smoking status (HR=1.97,95%CI 1.38-2.81, P<0.001). Cox analyses showed that a one standard deviation increase in HOMA-IR was associated with a 22% increase in the mortality after adjustment of potential confounders (HR=1.22, 95%CI 1.08-1.39, P=0.002). Conclusions: Insulin resistance is associated with increased risk of all-cause death in Chinese people without diabetes, suggesting that improving insulin resistance could be beneficial for people without diabetic in reducing risk of long-term all-cause mortality.

Effects of dietary corn germ meal levels on growth performance, serum biochemical parameters, meat quality, and standardized ileal digestibility of amino acids in Pekin ducks.

The study aimed to investigate the effects of dietary corn germ meal (CGM) levels on growth performance, carcass characteristic, serum biochemical indexes, meat physical and chemical quality, and standardized ileal digestibility of amino acids (SIDAA) in Pekin ducks from 10 to 42 d of age. A total of 420 ten-day-old Cherry Valley ducks were randomly allotted to 5 treatments with 6 replicate cages per treatment and 14 ducks per cages based on mean body weight. Five isonitrogenous and isocaloric experimental diets were formulated on a digestible amino acid basis to produce diets containing 0, 3, 6, 9, or 12% CGM. Results showed: 1) Compared with other groups, ducks fed 12% CGM significantly increased (P < 0.05) the feed to gain ratio. 2) Dietary CGM levels had no effect (P > 0.05) on the carcass traits and breast meat physical quality; but the content of crude protein presented a linear decrease (P < 0.05) in breast meat with increasing dietary CGM levels. 3) Serum biochemical indices (e.g., alanine aminotransferase, aspartate aminotransferase, glucose, high density lipoprotein cholesterol, total cholesterol, triglyceride, total protein, and urea) showed no significant differences among all groups (P > 0.05). 4) The levels of CGM had no significant effect on SIDAA of diets (P > 0.05), except for cysteine which showed a quadratic increase (P < 0.05). These results suggested that the optimal levels of CGM in diets for meat duck aged from 10 to 42 d should be below 9% based on feed to gain ratio and the content of crude protein in breast meat.

The impacts of egg storage time and maternal dietary vitamin E on the growth performance and antioxidant capacity of progeny chicks.

Two trials were designed to investigate the impacts of egg storage time and maternal dietary vitamin E (VE) supplementation on the growth performance and antioxidant capacity of progeny chicks. In total 512 Ross 308 broiler breeder hens (71-wk-old) were assigned to 2 dietary VE treatments (6 and 100 mg/kg) for 14 wk. Progeny chicks used in trials 1 and 2 were originated from eggs laid at week 10 (stored 0 d) and week 8 (stored 14 d), and week 14 (stored 0 d) and week 12 (stored 14 d), respectively. The 4 groups in trial 1 consisted of 2 levels of maternal VE (6 and 100 mg/kg) and 2 egg storage time (0 and 14 d). The 8 groups in trial 2 consisted of 2 levels of maternal VE (6 and 100 mg/kg), 2 egg storage time (0 and 14 d) and progeny sex (male and female). In trial 1, egg storage decreased the body weight, the liver total superoxide dismutase and total antioxidant capacity of 21-day-old offspring (P < 0.05), and the body weight gain and feed intake from 8 to 21 d and 1 to 21 d (P < 0.05); and increased the serum and liver malonaldehyde (MDA) of 7-day-old offspring and the ratio of feed: gain (F/G) from 1 to 7 d (P < 0.05). Maternal VE (100 vs. 6 mg/kg) decreased the F/G from 1 to 7 d and increased the serum total superoxide dismutase of 21-day-old offspring (P < 0.05). In trial 2, egg storage decreased the body weight of 42-day-old offspring, and the body weight gain and feed intake from 22 to 42 d and 1 to 42 d (P < 0.05); and increased the serum and liver MDA of 21- and 42-day-old offspring (P < 0.05). Maternal VE (100 vs. 6 mg/kg) reduced the serum MDA of 7-day-old offspring (P < 0.05). Interactively, maternal VE (100 vs. 6 mg/kg) reduced the serum MDA of offspring originated from stored eggs (P < 0.05), but not for that of offspring originated from unstored eggs in the two trials. It can be concluded that egg storage (14 vs. 0 d) decreased the growth performance and antioxidant capacity of offspring, while maternal dietary VE (100 vs. 6 mg/kg) supplementation could partly alleviate the reduction of antioxidant capacity (except for growth performance) of offspring induced by egg storage for the early phase post-hatch.

[Interpretation of the 2020 American society of colon and rectal surgeons clinical practice guidelines for the management of rectal cancer].

Located in the pelvic cavity and contiguous to the anal sphincter complex and urogenital organs, the rectum has more intricate anatomical features compared with the colon. Consequently, the treatment of rectal cancer involves more consideration, including pelvic radiation, lateral lymph node dissection, transanal access, postoperative function, sphincter preservation, and nonoperative management. Based on the last set of American society of colon and rectal surgeons (ASCRS) practice parameters for the management of rectal cancer published in 2013, the 2020 guidelines present evidence-based updates for both long-existing and emerging controversies on surgical management of rectal cancer. These updates include the indication for local resection, lymph node dissection for radical proctectomy, minimally invasive surgery, the "watch and wait" strategy for patients with clinical complete response, and prevention of anastomotic leak. Meanwhile, the guidelines recommend a risk-stratified approach for perioperative therapies for non-metastatic disease, and an individualized multimodality treatment based on treatment intent for synchronous metastatic disease.

Protease supplementation attenuates the intestinal health damage caused by low-protein diets in Pekin ducks.

The objective of this study was to investigate the effects of low-protein diets with low digestibility of feed ingredients on intestinal damage and to explore whether the protease supplementation can alleviate the damage in Pekin ducks. A total of 576 Pekin ducklings (6 replicate pens, 16 ducks/pen) were randomly assigned to 6 dietary treatments (3 × 2 factorial arrangement) in a randomized complete block design. Factors were CP levels (13.5%, 15.5%, and 17.5%) and protease (0 or 20,000U/kg). Compared with the diets containing 17.5% CP, low-protein diets (13.5% CP) showed suppressed (P < 0.05) growth performance and feed intake (FI); reduced (P < 0.05) serum-free arginine, isoleucine, leucine, methionine, phenylalanine, valine, and proline as well as the cecal acetate and propionate concentration; increased (P < 0.05) plasma and ileal mucosal tumor necrosis factor-α (TNF-α) concentration; and downregulated (P < 0.05) mRNA expression of TNF-α, nuclear transcription factor-κb, interferon gamma, and Occludin in ileal mucosa. Irrespective of the dietary CP levels, protease supplementation significantly increased (P < 0.05) the serum-free glutamic acid concentration while decreasing (P < 0.05) the plasma endotoxin, IL-6, and the cecal isovalerate concentration. A significant interactive effect was observed between low-protein diets and protease supplementation (P < 0.05) on serum-free arginine concentration, the ratio of ileal villus height to crypt depth, and the IL-6 concentration in ileal mucosa. These results indicated that low-protein diets could damage intestinal integrity to induce systemic inflammation response and at last to suppress growth performance. Protease supplementation could partly attenuate the negative effects on gut health caused by low-protein diets in Pekin ducks.

[Dilemma and strategy in surgery of chronic radiation intestinal injury].

Chronic radiation intestinal injury denotes the repeated and prolonged damage of intestine caused by radiotherapy to pelvic malignancy, which usually occurs after three months of radiotherapy. Surgical intervention is indicated when the progressive intestinal injury leads to the development of massive intestinal hemorrhage, obstruction, perforation, fistula and other late complications. However, there is no consensus on the surgical procedures. We illustrate the dilemma in surgical treatment from the points of pathological mechanism and the frequent sites of radiation intestinal injury. Meanwhile, we discuss the surgical alternatives of radiation intestinal injury based on the literature and our experience. The pathological mechanism of chronic radiation injury is progressive occlusive arteritis and parenchymal fibrosis. The frequently involved sites are distal ileum, sigmoid colon and rectum based on the radiotherapy region. The morbidity and mortality are high in surgery of chronic radiation injury due to poor ability of tissue healing, pelvic fibrosis, multiple organ damage, and poor physical condition. Definitive intestinal resection is one of the most common surgical procedures. Extended resection of diseased bowel to ensure that there is no radiation damage in at least one end of the anastomotic bowels is the key to decrease the risk of complications related to anastomotic sites.

Effects of microRNA-21 targeting PITX2 on proliferation and apoptosis of pituitary tumor cells.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Effects of microRNA-21 targeting PITX2 on proliferation and apoptosis of pituitary tumor cells, by M. Cui, M. Zhang, H.-F. Liu, J.-P. Wang, published in Eur Rev Med Pharmacol Sci 2017; 21 (13): 2995-3004-PMID: 28742208" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/13066.

An evaluation of direct PCR assays for the detection and quantification of Porphyromonas gingivalis.

Porphyromonas gingivalis has been linked to the development and progression of oesophageal squamous cell carcinoma (ESCC), and is considered to be a high-risk factor for ESCC. Currently, the commonly used methods for P. gingivalis detection are culture or DNA extraction-based, which are either time and labour intensive especially for high-throughput applications. We aimed to establish and evaluate a rapid and sensitive direct quantitative polymerase chain reaction (qPCR) protocol for the detection of P. gingivalis without DNA extraction which is suitable for large-scale epidemiological studies. Paired gingival swab samples from 192 subjects undergoing general medical examinations were analysed using two direct and one extraction-based qPCR assays for P. gingivalis. Tris-EDTA buffer-based direct qPCR (TE-direct qPCR), lysis-based direct qPCR (lysis-direct qPCR) and DNA extraction-based qPCR (kit-qPCR) were used, respectively, in 192, 132 and 60 of these samples for quantification of P. gingivalis. The sensitivity and specificity of TE-direct qPCR was 95.24% and 100% compared with lysis-direct qPCR, which was 100% and 97.30% when compared with kit-qPCR; TE-direct qPCR had an almost perfect agreement with lysis-direct qPCR (κ = 0.954) and kit-qPCR (κ = 0.965). Moreover, the assay time used for TE-direct qPCR was 1.5 h. In conclusion, the TE-direct qPCR assay is a simple and efficient method for the quantification of oral P. gingivalis and showed high sensitivity and specificity compared with routine qPCR.

[Identification and functional study of the Schistosoma japonicum epidermal growth factor receptor gene].

OBJECTIVE: To characterize the epidermal growth factor receptor (EGFR) gene in Schistosoma japonicum (SjEGFR gene) and investigate the role of the EGFR gene in regulating the growth, reproductive system, maturation and fecundity of S. japonicum. METHODS: Rapid amplification of cDNA ends (RACE) was performed to obtain the full length of the SjEGFR gene, and the SjEGFR gene expression was quantified in different developmental stages of S. japonicum using a quantitative real-time PCR (qPCR) assay. The tissue localization of the SjEGFR gene was detected in 22-day parasite using whole-mount in situ hybridization (WISH). Following RNA interference (RNAi)-induced knockdown of the SjEGFR gene, the worm length, pairing rate and worm burden of S. japonicum were measured, and the worm morphology was observed using optical microscopy and confocal microscopy. RESULTS: The SjEGFR gene was identified with a conserved tyrosine-kinase active site, and the SjEGFR gene expression was detected at various developmental stages in male and female parasites. WISH showed that the transcript of the SjEGFR gene was localized on the tegument and in the digestive organs of S. japonicum. RNAi-induced SjEGFR knockdown resulted in marked suppression of the worm growth, smaller size of male testicles that contained more immature spermatocytes, and apparent impairment of ovary and vitelline gland development. In addition, no eggs were found in the uterus of SjEGFR knocked-down female parasites, indicating the interruption of egg production. CONCLUSIONS: Inhibition of SjEGFR expression may remarkably suppress the growth and maturation of S. japonicum, and interrupt the egg production.

MiR-142-5p reverses the resistance to gefitinib through targeting HOXD8 in lung cancer cells.

OBJECTIVE: To investigate the role and potential mechanism of micro ribonucleic acid (miR)-142-5p in the acquired resistance to gefitinib in lung cancer cells. MATERIALS AND METHODS: The drug resistance of PC9/G cells was detected via methyl thiazolyl tetrazolium (MTT) assay. Expression levels of miR-142-5p and HOXD8 in PC9 and PC9/G cells were detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and Western blotting. PC9/G cells were transfected with miR-142-5p mimic, while PC9 cells were transfected with miR-142-5p inhibitor. Subsequently, expression changes of HOXD8 were determined using qRT-PCR and Western blotting, cell sensitivity to gefitinib was detected through MTT assay, and the apoptosis was detected via flow cytometry. Moreover, Dual-Luciferase reporter assay was conducted to determine the relationship between HOXD8 and miR-142-5p. Finally, potential involvement of HOXD8 in miR-142-5p-regulated gefitinib sensitivity was confirmed via rescue tests. RESULTS: PC9/G cells were more significantly resistant to gefitinib compared with its parental PC9 cells. MiR-142-5p was down-regulated in PC9/G cells, while that of HOXD8 was up-regulated in PC9/G cells. Transfection of miR-142-5p mimic could inhibit the expression level of HOXD8 in PC9/G cells and reverse its resistance to gefitinib. Conversely, transfection of miR-142-5p inhibitor could upregulate HOXD8 in PC9 cells and promote its resistance to gefitinib. According to the Dual-Luciferase reporter assay, miR-142-5p could suppress the expression of HOXD8 through the targeted binding to the HOXD8 3'UTR. Moreover, miR-142-5p could regulate mitochondrial apoptosis pathway by targeting HOXD8. Finally, rescue tests confirmed that miR-142-5p regulated the sensitivity of PC9 cells to gefitinib by acting on the target gene HOXD8. CONCLUSIONS: Down-regulation of miR-142-5p induces the resistance of lung cancer PC9 cells to gefitinib by upregulating HOXD8.

MiR-129-5p inhibits proliferation of gastric cancer cells through targeted inhibition on HMGB1 expression.

OBJECTIVE: The aim of this study was to investigate the effects of micro ribonucleic acid (miR)-129-5p on the proliferation and apoptosis of gastric cancer cells via targeted repression on the expression of high mobility group protein B1 (HMGB1). PATIENTS AND METHODS: Expression levels of miR-129-5p and HMGB1 in gastric cancer tissues (n=25) and adjacent normal tissues were measured via reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The regulatory effect of miR-129-5p on the proliferation of gastric cancer MGC-803 and SGC7901 cells was determined through Cell Counting Kit-8 (CCK-8) assay. Flow cytometry was employed to analyze the apoptosis rate of gastric cancer cells. To further discover the mechanism of miR-129-5p in regulating malignant behaviors of gastric cancer cells, the miRDB database was employed to predict the binding targets of miR-129-5p. Finally, binding sites of HMGB1 3'-untranslated region (3'-UTR) to miR-129-5p were discovered. Subsequently, HMGB1 wild-type or mutant 3'-UTR Luciferase reporter vectors were constructed, and transfected to MGC-803 and SGC7901 cells together with miR-129-5p or negative control miRNA. Next, Western blotting was adopted to measure the protein expression level of HMGB1 in MGC-803 and SGC7901 cells transfected with miR-129-5p or negative control miRNA, so as to investigate whether miR-129-5p affected HMGB1 protein expression. Additionally, to determine whether HMGB1 mediated the regulatory effect of miR-129-5p on the proliferation of gastric cancer cells, MGC-803 and SGC7901 cells were transfected with pcDNA-HMGB1 or pcDNA-vector, respectively. The expression level of HMGB1 was measured via RT-qPCR, and cell proliferation was determined by CCK-8 assay. RESULTS: The expression level of miR-129-5p in gastric cancer tissues was significantly lower than that in adjacent normal tissues (p<0.001). Meanwhile, the level of miR-129-5p was overtly lower in gastric cancer MGC-803 and SGC7901 cell lines than that in normal gastric mucosal epithelial GES-1 cells (p<0.001). These results indicated that miR-129-5p was lowly expressed in gastric cancer tissues and cell lines. Subsequent results demonstrated that the expression of HMGB1 increased remarkably in gastric cancer tissues compared with normal adjacent tissues (p<0.05). The proliferation ability of MGC-803 (p<0.001) and SGC7901 (p<0.01) cells with over-expressed miR-129-5p was remarkably weakened. Overexpression of miR-129-5p distinctly promoted the apoptosis rate of gastric cancer MGC-803 (p<0.01) and SGC7901 (p<0.001) cells. Moreover, miR-129-5p up-regulation significantly reduced the Luciferase activity of wild-type HMGB1 (p<0.001). However, no significant effect was observed on that of mutant HMGB1. The results suggested that overexpression of miR-129-5p significantly down-regulated the level of HMGB1 in gastric cancer cells. In addition, the messenger RNA (mRNA) level of HMGB1 in cells transfected with miR-129-5p also decreased significantly (p<0.001). HMGB1 overexpression overtly reversed the inhibitory effect of miR-129-5p on the proliferation of gastric cancer cells (p<0.05). All these results demonstrated that the miR-129-5p/HMGB1 axis played a key role in regulating the growth of gastric cancer cells. CONCLUSIONS: MiR-129-5p suppresses the progression of gastric cancer through targeted inhibition on the expression of HMGB1.

Evaluation of the standardized ileal digestibility of amino acids of rapeseed meals varying in protein solubility for Pekin ducks.

This study was conducted to determine whether protein solubility (PS) of rapeseed meals (RSM) can affect standardized ileal amino acid digestibility (SIDAA) in meat ducks. A total of 1,168, 14-days-old ducks were randomly allotted to 23 treatments (6 cages per diet, 8 ducks per cage) and 1 nitrogen-free diet treatment (8 cages, 8 ducks per cage) based on body weight. The 23 experimental diets consisted of a corn-soybean meal basal diet, and 22 diets containing 15% RSM: 85% basal diet. Titanium dioxide (0.5%) was included in all diets as an indigestible marker. On day 18, all ducks were euthanized by carbon dioxide asphyxiation and digesta samples from the ileum. The contents of PS, ether extract (EE), glucosinolate, isothiocyanate, and oxazolidine were significantly different (P < 0.05) in the 22 RSM, with the CV being 52.62, 49.23, 86.84, 90.19, and 81.98%, respectively. The content of lysine (Lys) and methionine in the 22 RSM samples ranged from 1.03 to 2.71% (CV 24.19%) and from 0.33 to 0.65% (CV 15.17%), respectively. The SIDAA, except for leucine (Leu) and tyrosine, of the 22 RSM samples varied significantly (P < 0.05). A positive correlation was observed (P < 0.05) between PS and standardized ileal digestibility (SID) of Lys, isoleucine, valine, phenylalanine, histidine, serine, cysteine, and tyrosine. The R2 value of multiple linear regression equations for predicting the SID of amino acids (AA) was best for Lys (R2 = 0.958 using dry matter, crude protein, EE, crude fiber, acid detergent fiber, and PS) and least significant for Leu (R2 = 0.348 using crude fiber and ash) with intermediate values for other AA (R2 = 0.359-0.837, P < 0.05). These results suggest that PS varying from 15.06 to 98.08%, also varied considerably in the proximate nutrient content, AA composition, and antinutritional factor content, which was reflected in considerable differences in the duck's SID of AA in RSM. Therefore, PS value can partly reflect the quality of RSM.