Probiotics are natural systems bridging synthetic biology, physical biotechnology, and immunology, initiating innate and adaptive anti-tumor immune activity. We previously constructed an all-in-one engineered food-grade probiotic Lactococcus lactis (FOLactis) which could boost the crosstalk among different immune cells such as dendritic cells (DCs), natural killer cells, and T cells. Herein, considering the limited clinical efficacy of naked personalized neoantigen peptide vaccines, we decorate FOLactis with tumor antigens by employing a Plug-and-Display system comprising membrane-inserted peptides. Intranodal injection of FOLactis coated with neoantigen peptides (Ag-FOLactis) induces robust DCs presentation and neoantigen-specific cellular immunity. Notably, Ag-FOLactis not only triggers a 45-fold rise in the quantity of locally reactive neoantigen-specific T cells but also induces epitope spreading in both subcutaneous and metastatic tumor-bearing models, leading to potent inhibition of tumor growth. These findings imply that Ag-FOLactis represents a powerful platform to rapidly and easily display antigens, facilitating the development of a bio-activated platform for personalized therapy.

Objective:To evaluate the short-term efficacy of parallel overlapping anastomosis in 3D laparoscopic radical resection for transverse colon cancer.Methods:A retrospective analysis of clinical data was conducted on 73 patients who underwent 3D laparoscopic radical resection and parallel overlapping anastomosis for reconstruction of digestive tract transverse colon cancer at He'nan Provincial People's Clinical Medical School of Zhengzhou University between Apr 2019 and Jul 2023.Results:The average operation time was (165.3±28.9) min, the parallel overlapping anastomosis time was (15.6±2.6) min, the intraoperative blood loss was (47.7±19.4) ml, the first postoperative exhaust time was(2.2±0.9) d, the first time getting out of bed was (1.1±0.4) d, the number of lymph node dissection was (18.2±5.1), the first postoperative oral fluid diet time was (4.4±0.8) d, the postoperative hospital stay was (6.4±1.8) d. Anastomotic leakage occurred in one patient , who was discharged after enterostomy; Two patients with poor wound healing were cured after wound debridement and dressing change; Pulmonary infection in one patient was healed by anti-infection treatment. The postoperative complication rate was 6%, and no complications such as anastomotic bleeding, anastomotic stenosis, intestinal obstruction, abdominal infection occurred.Conclusion:Parallel overlapping anastomosis is a safe and reliable anastomotic procedure for the reconstruction of digestive tract in patients undergoing 3D laparoscopic radical resection of transverse colon cancer.

Objective To explore the regulatory role of N-methyl berbamine(N-MB)in intracellular calcium homeostasis in H9c2 car-diomyocytes,and,thereby,clarify the possible mechanism of the myocardial protective effect of N-MB.Methods Binding of N-MB to CaV1.2 channels was simulated using the MOE software,and the binding affinity and binding mode were determined.The hCaV1.2 gene was transfected into HEK293 cells,and the effect of N-MB(30 μmol/L)on the CaV1.2 current was detected using the whole-cell patch clamp technique.In addition,a Fluo 3-AM fluorescent probe was loaded into H9c2 cardiomyocytes,and the effect of N-MB(3,30 μmol/L)on intracellular calcium ion concentration was observed under a laser confocal microscope.The effect of N-MB(3,30 μmol/L)on the expression of Ca2+regulation-related genes Cacna1c,Cacnb2,Ryr2,Serca2a,and Ncx1 in H9c2 cardiomyocytes was examined using real-time quantitative PCR.Results N-MB was predicted to bind to CaV1.2 channels.The binding sites mainly involved Phe1191,Thr1420,and Asn771,and the binding modes were H-donor,pi-pi,and pi-H.N-MB(30 μmol/L)significantly inhibited CaV1.2 currents,with an inhibition rate of 76.09％±7.41％.The fluorescence intensity of intracellular Ca2+level in H9c2 cardiomyocytes was significantly enhanced with N-MB treatment(3,30 μmol/L,P＜0.01).Compared with the control group,differences in the expression of Cacna1c,Serca2a,and Ncx1 in H9c2 cardiomyocytes were not significant after N-MB(3,30 μmol/L)intervention(P＞0.05),whereas the expression of Cacnb2 significantly reduced(P＜0.001)and the expression of Ryr2 significantly increased(P＜0.05).Conclusion N-MB binds to CaV1.2 calcium channels.N-MB may regulate intracellular calcium homeostasis by inhibiting calcium currents by decreasing the gene expression of CaV1.2 calcium channels.Additionally,N-MB may also increase intracellular Ca2+concentration by promoting the expression of Ryr2,which could be the mechanism underlying the myocardial protective effect of N-MB.

Objective To explore the regulatory role of N-methyl berbamine(N-MB)in intracellular calcium homeostasis in H9c2 car-diomyocytes,and,thereby,clarify the possible mechanism of the myocardial protective effect of N-MB.Methods Binding of N-MB to CaV1.2 channels was simulated using the MOE software,and the binding affinity and binding mode were determined.The hCaV1.2 gene was transfected into HEK293 cells,and the effect of N-MB(30 μmol/L)on the CaV1.2 current was detected using the whole-cell patch clamp technique.In addition,a Fluo 3-AM fluorescent probe was loaded into H9c2 cardiomyocytes,and the effect of N-MB(3,30 μmol/L)on intracellular calcium ion concentration was observed under a laser confocal microscope.The effect of N-MB(3,30 μmol/L)on the expression of Ca2+regulation-related genes Cacna1c,Cacnb2,Ryr2,Serca2a,and Ncx1 in H9c2 cardiomyocytes was examined using real-time quantitative PCR.Results N-MB was predicted to bind to CaV1.2 channels.The binding sites mainly involved Phe1191,Thr1420,and Asn771,and the binding modes were H-donor,pi-pi,and pi-H.N-MB(30 μmol/L)significantly inhibited CaV1.2 currents,with an inhibition rate of 76.09％±7.41％.The fluorescence intensity of intracellular Ca2+level in H9c2 cardiomyocytes was significantly enhanced with N-MB treatment(3,30 μmol/L,P＜0.01).Compared with the control group,differences in the expression of Cacna1c,Serca2a,and Ncx1 in H9c2 cardiomyocytes were not significant after N-MB(3,30 μmol/L)intervention(P＞0.05),whereas the expression of Cacnb2 significantly reduced(P＜0.001)and the expression of Ryr2 significantly increased(P＜0.05).Conclusion N-MB binds to CaV1.2 calcium channels.N-MB may regulate intracellular calcium homeostasis by inhibiting calcium currents by decreasing the gene expression of CaV1.2 calcium channels.Additionally,N-MB may also increase intracellular Ca2+concentration by promoting the expression of Ryr2,which could be the mechanism underlying the myocardial protective effect of N-MB.

Objective:To evaluate the short-term efficacy of parallel overlapping anastomosis in 3D laparoscopic radical resection for transverse colon cancer.Methods:A retrospective analysis of clinical data was conducted on 73 patients who underwent 3D laparoscopic radical resection and parallel overlapping anastomosis for reconstruction of digestive tract transverse colon cancer at He'nan Provincial People's Clinical Medical School of Zhengzhou University between Apr 2019 and Jul 2023.Results:The average operation time was (165.3±28.9) min, the parallel overlapping anastomosis time was (15.6±2.6) min, the intraoperative blood loss was (47.7±19.4) ml, the first postoperative exhaust time was(2.2±0.9) d, the first time getting out of bed was (1.1±0.4) d, the number of lymph node dissection was (18.2±5.1), the first postoperative oral fluid diet time was (4.4±0.8) d, the postoperative hospital stay was (6.4±1.8) d. Anastomotic leakage occurred in one patient , who was discharged after enterostomy; Two patients with poor wound healing were cured after wound debridement and dressing change; Pulmonary infection in one patient was healed by anti-infection treatment. The postoperative complication rate was 6%, and no complications such as anastomotic bleeding, anastomotic stenosis, intestinal obstruction, abdominal infection occurred.Conclusion:Parallel overlapping anastomosis is a safe and reliable anastomotic procedure for the reconstruction of digestive tract in patients undergoing 3D laparoscopic radical resection of transverse colon cancer.

Objectives:To investigate the application value of laparoscopic double stapler firings and double stapling technique combined with rectal eversion and total extra-abdominal resection (LDER) in the anal preservation treatment of low rectal cancer.Methods:Inclusion criteria: (1) age was 18-70; (2) the distance of the lower tumor edge from the anal verge was 4-5 cm; (3) primary tumor with a diameter ≤3 cm; (4) preoperative staging of T1~2N1~2M0; (5) "difficult pelvis", defined as ischial tuberosity diameter<10 cm or body mass index>25 kg/m 2; (6) patients with strong intention for sphincter preservation; (7) no preoperative treatment (e.g., chemotherapy, radiotherapy, molecular targeted therapy, or immunotherapy); (8) no lateral lymph node enlargement; (9) no previous anorectal surgery; (10) patients with good basic condition who could tolerate surgery. Exclusion criteria: (1) previously suffered from malignant tumors of the digestive tract or currently suffering from malignant tumors out of the digestive tract; (2) patients with preoperative anal dysfunction (Wexner score ≥ 10), or fecal incontinence. The specific surgical steps are as follows: the distal end of the rectum was dissected to the level of the interspace between internal and external sphincters of anal canal. Five centimeters proximal to the tumor, the mesorectum was ligated, and a liner stapler was used to transect the rectum. The distal rectum with the tumor were then everted and extracted through the anus. The rectum was transected 0.5-1.0 cm distal to the tumor with a linear stapler. Full thickness suture was used to reinforce the stump of the rectum, which was then brought back into the pelvic cavity. Finally, an end-to-end anastomosis between the colon and the rectum was performed. A retrospective descriptive study was performed of the clinical and pathological data of 12 patients with T1-T2 stage low rectal cancer treated with LDER at Henan Provincial People's Hospital from January 2020 to December 2022. Results:All 12 patients successfully completed LDER with sphincter preservation, without conversion to open surgery or changes in surgical approach. The median surgical time was 272 (155-320) minutes, with a median bleeding volume of 100 (50-200) mL. No protective stoma was performed, and all patients received R0 resection. The average hospital stay was 9 (7-15) days. There were no postoperative anastomotic leakage or perioperative deaths. All 12 patients received postoperative follow-up, with a median follow-up of 12 months (6-36 months) and a Wexner score of 8 (5-14) at 6 months postoperatively. There was no tumor recurrence or metastasis during the follow-up period.Conclusions:LDER is safe and effective for the treatment of low rectal cancer.

Objective:To investigate the expression and clinical application value of exosomal circRPS6 in serum of colorectal cancer (CRC) patients.Methods:Peripheral serum samples were collected from 115 CRC patients admitted to Henan Provincial People′s Hospital from January 2019 to December 2020. There were 68 males and 47 females, aged (63.0±9.5) years. Meanwhile, one hundred and twenty healthy subjects from the same period wereenrolled, with 70 males and 50 females, aged (61.0±10.7) years. In addition, sixty pairs of tumor and adjacent tissue specimens from CRC patients undergoing surgical treatment were collected. The circRPS6 expression in serum exosome and tissue of CRC patients were detected via real-time fluorescence quantitative PCR (RT-qPCR), and its relationship with clinicopathological features and prognosis of CRC patients were also investigated. The levels of CEA and CA19-9 in serum were detected by electrochemiluminescence assay. The ROC curve and AUC were used to estimate the diagnostic capacity. Univariate and multivariate regression analysis was performed using Cox proportional hazard analysis.Results:The expression level of circRPS6 in CRC tissue was significantly higher than that in adjacent tissue( Z=5.38, P<0.001). Compared with healthy control, the expression of serum exosomal circRPS6 was significantly upregulated in the CRC group( t=14.52, P<0.001). ROC curve analysis results showed that the AUC of exosomal circRPS6 was 0.882, which had a higher diagnostic efficacy in CRC patients than CEA and CA19-9 detection. There was a positive correlation between the expression level of exosomal circRPS6 with TNM stage and lymph node metastasis and distant metastasis( P<0.05). Kaplan-Meier survival analysis revealed that CRC patients with low exosomal circRPS6 levels had a much longer average survival time compared with those in high group. Moreover,multivariate analysis results indicated that exosomal circRPS6 was an independent prognostic factor in colorectal cancer. Conclusion:Exosomal circRPS6 is highly expressed in the serum of CRC patients and correlated with malignant progression and poor prognosis, which is expected to be a potential marker for the diagnosis and prognosis evaluation of CRC patients.

Objectives:To investigate the application value of laparoscopic double stapler firings and double stapling technique combined with rectal eversion and total extra-abdominal resection (LDER) in the anal preservation treatment of low rectal cancer.Methods:Inclusion criteria: (1) age was 18-70; (2) the distance of the lower tumor edge from the anal verge was 4-5 cm; (3) primary tumor with a diameter ≤3 cm; (4) preoperative staging of T1~2N1~2M0; (5) "difficult pelvis", defined as ischial tuberosity diameter<10 cm or body mass index>25 kg/m 2; (6) patients with strong intention for sphincter preservation; (7) no preoperative treatment (e.g., chemotherapy, radiotherapy, molecular targeted therapy, or immunotherapy); (8) no lateral lymph node enlargement; (9) no previous anorectal surgery; (10) patients with good basic condition who could tolerate surgery. Exclusion criteria: (1) previously suffered from malignant tumors of the digestive tract or currently suffering from malignant tumors out of the digestive tract; (2) patients with preoperative anal dysfunction (Wexner score ≥ 10), or fecal incontinence. The specific surgical steps are as follows: the distal end of the rectum was dissected to the level of the interspace between internal and external sphincters of anal canal. Five centimeters proximal to the tumor, the mesorectum was ligated, and a liner stapler was used to transect the rectum. The distal rectum with the tumor were then everted and extracted through the anus. The rectum was transected 0.5-1.0 cm distal to the tumor with a linear stapler. Full thickness suture was used to reinforce the stump of the rectum, which was then brought back into the pelvic cavity. Finally, an end-to-end anastomosis between the colon and the rectum was performed. A retrospective descriptive study was performed of the clinical and pathological data of 12 patients with T1-T2 stage low rectal cancer treated with LDER at Henan Provincial People's Hospital from January 2020 to December 2022. Results:All 12 patients successfully completed LDER with sphincter preservation, without conversion to open surgery or changes in surgical approach. The median surgical time was 272 (155-320) minutes, with a median bleeding volume of 100 (50-200) mL. No protective stoma was performed, and all patients received R0 resection. The average hospital stay was 9 (7-15) days. There were no postoperative anastomotic leakage or perioperative deaths. All 12 patients received postoperative follow-up, with a median follow-up of 12 months (6-36 months) and a Wexner score of 8 (5-14) at 6 months postoperatively. There was no tumor recurrence or metastasis during the follow-up period.Conclusions:LDER is safe and effective for the treatment of low rectal cancer.

Objective:To investigate the clinical efficacy of three-dimensional (3D) laparos-copic radical resection of rectal cancer with left colic artery preservation and natural orfice specimen extraction surgery (NOSES).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 186 patients with rectal cancer who were admitted to Henan Provincial People's Hospital from December 2018 to December 2019 were colleted. There were 120 males and 66 females, aged from 30 to 81 years, with a median age of 59 years. Patients underwent 3D laparoscopic radical resection of rectal cancer. Observation indicators: (1) surgical and postoperative situations; (2) follow-up. Follow-up using outpatient examination or telephone interview was conducted to detect survival of patients and tumor recurrence up to January 2021. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers and percentages. Result:(1) Surgical situations: 186 patients with rectal cancer under-went 3D laparoscopic radical resection and postoperative with left colic artery preservation and NOSES. The operation time was (123±24)minutes, volume of intraoperative blood loss was (30±20)mL, the numer of lymph nodes dissected was 15±7. The time to postoperative first flatus, time to semiliquid food intake, time to postoperative out-of-bed activities were (2.3±0.7)days, (4.1±1.4)days, (2.9±1.0)days, respectively. The incidence of postoperative complications was 8.06%(15/186). The duration of postoperative hospital stay of 186 patients was (6.6±1.9)days and the treatment cost was (3.8±1.1) ten thousand yuan. Results of postoperative pathological examination showed 54 cases of low differentiated adenocarcinoma, 97 cases of moderate differentiated adenocarcinoma, 19 cases of high differentiated adenocarcinoma and 16 cases of mucinous adenocarcinoma. (2) Follow-up: 186 patients with rectal cancer were followed up for 13 to 24 months, with a median follow-up time of 13 months. During the follow-up, 18 patients had tumor recurrence or metastasis including 6 patients of death, 168 cases recovered well.Conclusion:3D laparoscopic radical resection of rectal cancer with left colic artery preservation and NOSES is safe and feasible.

Objective:To investigate the expression of circular ribonucleic acid ABCB10 (circABCB10) in colorectal cancer tissues and cells and its effects on cell biological behavior, radiosensitivity and growth of subcutaneous xenografts.Methods:The tumor tissue and adjacent tissue from colorectal cancer patients treated in Henan People′s Hospital were collected from January 2018 to December 2018. Quantitative polymerase chain reaction (qPCR) was used to detect the expressions of circABCB10 and miR-217, cell viability was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT), cell apoptosis rate was detected by flow cytometry, cell migration and invasion were detected by Transwell method, cell radiosensitivity was detected by colony formation assay. The downstream miRNAs of circABCB10 were predicted by Circular RNA Interactome and verified by the dual luciferase reporter gene experiment. The effect of circABCB10 on the growth of transplanted tumor was examined in nude mice.Results:The expression level of circABCB10 mRNA in colorectal cancer tissues was (3.97±2.12), higher than (1.13±0.64) in adjacent tissues ( P<0.05). The expression level of circABCB10 mRNA in FHC cells was (1.00±0.09), lower than that (4.53±0.44) in SW480, (3.12±0.32) in HCT116 and (3.51±0.36) in HT29 cells, respectively (all P<0.05). The MTT results showed that the absorbance values of SW480 cells in si-circABCB10-1 group at 48 and 72 hours after transfection were (0.36±0.04) and (0.43±0.04), lower than (0.48±0.05) and (0.82±0.08) in circ-negative control (NC) group, respectively (all P<0.05). The number of migrating cells and invasive cells in si-circABCB10-1 group were (45±8) and (34±7), lower than (106±21) and (84±15) in circ-NC group, respectively (all P<0.01). The radiosensitization ratio was 1.632. The results of subcutaneous transplantation assay showed that the tumor volume and tumor weight of the si-circABCB10-1 group were significantly lower than circ-NC group after 8 days of inoculation ( all P<0.05). MiR-217 is a target gene of circABCB10. Inhibition of miR-217 reversed the inhibitory effect of circABCB10 silencing on cell proliferation, migration, invasion and subcutaneous xenograft growth in nude mice and the radiosensitization activity. Conclusion:Silence of circABCB10 can up-regulate the expression of miR-217 to inhibit the proliferation, migration, invasion and growth of subcutaneous xenografts and increase the radiosensitivity of SW480 cells, which reveals the underlying molecular mechanism of colorectal cancer progression and provides a new sensitizing target for clinical radiotherapy of colorectal cancer.