Objective To evaluate the effect of neurotrophin-3 (NT-3) on the expression of serine/threonine protein kinase (Akt) during ropivacaine-induced neurotoxicity to the spinal cord of rats.Methods Healthy adult male Sprague-Dawley rats,aged 1-2 months,weighing 280-320 g,were used in the study.A catheter was inserted at L5,6 interspace into the epidural space of rats.A total of 108 rats,in which intrathecal catheters were successfully implanted,were randomly divided into 3 groups (n =36each):control group (group C),1％ ropivacaine group (group R),and 1％ ropivacaine + NT-3 group (group NT).The equal volume of normal saline was given in group C,1％ ropivacaine 0.12 ml/kg was injected via the intrathecal catheter once every 1.5 h for 8 times in total in R and NT groups.In addition,NT-3 0.1 mg/kg was simultanenously injected via the intrathecal catheter in group NT.On days 1,3,5,7,14 and 28 after the end of administration (T1-6),6 rats were sacrificed in each group.Their lumbar enlargements were removed for determination of neuronal apoptosis (using TUNEL) and Akt expression (by immuno-histochemistry).The apoptotic rate was calculated.Results Compared with group C,the apoptotic rate was significantly increased at T1-4,and Akt expression was significantly up-regulated at T1-3 in group R,and the apoptotic rate were significantly increased,and Akt expression was significantly up-regulated at T1-3 in group NT (P＜0.05).Compared with group R,the apoptotic rate was significantly decreased at T3,4,and Akt expression was significantly down-regulated at T2.3 in group NT (P＜0.05).Conclusion The mechanism by which NT-3 reduces ropivacaine-induced neurotoxicity to the spinal cord may be related to down-regulation of the expression of Akt in rats.

AIM: To detect the role of cyclic nucleotides in the alleviation of hypoxic pulmonary vasoconstriction (HPV) in chronic hypoxic animals. METHODS: The intracellular cAMP and cGMP of the cultured porcine pulmonary arterial smooth muscle cells (PASMC) and endothelial cells (PAEC) were assayed by RIA. The length of single PASMC during acute hypoxia was measured by imaging analysis system. RESULTS: The basal levels of cAMP and cGMP in PASMC and cGMP in PAEC of Chronic hypoxic groups decreased remarkably compared with normoxic groups ( P

Objective:To investigate the role of LncRNA-TUG1 in the injury of intestinal epithelial cells induced by lipopolysaccharide (LPS).Methods:LPS was used to treat HIEC-6 human intestinal epithelial cells for 24 h to construct a sepsis injury model. Whole transcriptome RNA sequencing was used to analyze the expression changes of mRNA, microRNA and lncRNA in HIEC-6 cells after LPS treatment. Real-time fluorescence quantitative (qRT-PCR) and Western blot was performed to detect the expression changes of lncRNA-TUG1, microRNA-132-3p (miR-132-3p), SIRT1 mRNA and SIRT1 protein in HIEC-6 cells after LPS treatment. The expression levels of LncRNA-TUG1, miR-132-3p and SIRT1 were artificially changed by in vitro transfection. qRT-PCR and Western blot were used to confirm the regulatory effect of lncRNA-TUG1 on microRNA-132-3p and SIRT1. CCK-8 and flow cytometry were used to analyze the effects of LncRNA-TUG1, miR-132-3p and SIRT1 on the proliferation and apoptosis of HIEC-6 cells. The dual luciferase report analysis was used to verify the targeting relationship between LncRNA-TUG1, miR-132-3p and SIRT1. Statistical analysis was performed using SPSS 17.0, and differences between the two groups were compared using independent sample t test. Results:RNA sequencing results showed that the expressions of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells after LPS treatment ( t=3.26, P<0.05 and t=2.55, P<0.05), but the expression of miR-132-3p was increased ( t=4.12, P<0.05). In vitro cell experiments, the expression of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells treated with LPS ( t=5.69, P<0.05 and t=5.712, P<0.05), while the expression of miR-132-3p was increased ( t=3.88, P<0.05). Overexpression of lncRNA-TUG1 increased the proliferation rate ( t=6.55, P<0.05) and decreased the apoptosis rate ( t=3.94, P<0.05) of LPS-treated cells. Upregulation of lncRNA-TUG1 decreased the expression of miR-132-3p ( t=4.66, P<0.05), and increased the mRNA and protein levels of SIRT1 ( t=3.91, P<0.05). Transfection of miR-132-3P mimic could inhibit the mRNA ( t=4.08, P<0.05) and protein levels of SIRT1. In LPS-treated cells, the cells co-transfected with miR-132-3pmimic and siRNA-SIRT1 had a lower proliferation rate ( t=4.55, P<0.05 and t=5.67, P<0.05) and a higher apoptosis rate ( t=3.90, P<0.05 and t=4.22, P<0.05) than those transfected with only pcDNA3.1-lncRNA-TUG. Conclusions:lncRNA-TUG1 may act as a ceRNA to regulate miR-132-3p/SIRT1, therefore alleviating HIEC-6 cell injury caused by LPS. Intervention of lncRNA-TUG1/miR-132-3p/SIRT1 regulatory pathway may become a potential strategy to prevent sepsis-induced intestinal mucosal damage.

This research focuses on the application of multiple interactive modes in online teaching, combined with the actual teaching cases of the anesthesia equipment course of Xiangya Anesthesiology Specialty of Central South University, showing in detail the preparations for interactive teaching before anesthesia equipment learning, the interaction in online classrooms, the extension of interactive teaching outside the classroom, and the evaluation of interactive teaching feedback mechanism throughout the implementation process. By establishing a "host-guest-viewer" mode, the effect of online live broadcasting is maximized. Through the 360-degree materialized explanation with students as the main body, we will make opening in the pain points and blocking points of online teaching in which students do not go to class and students have no thinking, and promote the improvement of online teaching quality and efficiency. In the following practice, we must continue to work on issues such as the improvement of teacher talent quality, the building of an efficient talent team, and the construction of practical application value evaluation systems for teaching.

OBJECTIVE: To investigate the effect of pretreatment with neurotrophin-3 (NT-3) on intrathecal ropivacaine in rats. METHODS: A total of 144 male Sprague Dawley rats weighing 280-320 g were successfully implanted with microspinal cather following the improved methods of Yaksh. The rats were randomly divided into 4 groups and given saline (Group NS, n=36), 0.5% ropivacaine (Group M, n=36), 1% ropivacaine (Group R, n=36), and ropivacaine+NT-3 (Group T, n=36). The rats received 0.12 mL/ kg body weight of ropivacaine at 0.5% or 1%, or normal saline only, via an implanted intrathecal catheter at 90-min interval for 12 h in Group NS, M, R and T. In the meantime the rats also received NT-3 0.1 mg/kg in group T. On days 1, 3, 5, 7, 14 and 28, we assessed the paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL), behavioural change and histopathological damage score changed for possible neuronal injury within the spinal cord. RESULTS: Compared with Group NS and Group M, the PWMT and PWTL were significantly higher on 1, 3, 5 d and the histopathological damage score was significantly higher on 1, 3, 5, 7, 14 d in Group R (P<0.05). Compared with Group T, the PWMT and PWTL in Group R were significantly higher on 1, 3, 5 d and histopathological damage score was significantly higher on 5, 7, 14 d (P<0.05). CONCLUSION NT-3 pretreatment in mice has obvious protective effect against repeated intrathecal injection of 1% ropivacaine in the spinal nerve.
Amides ; adverse effects ; Animals ; Injections, Spinal ; Male ; Neurotrophin 3 ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Ropivacaine ; Spinal Cord ; drug effects

Objective:To explore the expression of microRNA-4695-5p in the serum of colorectal cancer patients and its effect on the proliferation and invasion of colorectal cancer CACO-2 cells.Methods:A total of 43 serum samples of colorectal cancer patients who were admitted to the Department of Gastrointestinal Surgery, Luoyang Central Hospital Affiliated to Zhengzhou University from March 2018 to November 2021 were selected, and serum samples of 43 healthy people who underwent outpatient physical examination were used as controls. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels in the serum of colorectal cancer patients and those of healthy individuals. The miR-4695-5p overexpression plasmid or the negative control plasmid were transfected into CACO-2 cells, namely the miR-4695-5p group and the control group, and the transfection efficiency was verified by qRT-PCR. CCK8 method and Transwell experiment were used to detect the effect of overexpression of miR-4695-5p on the proliferation and invasion of CACO-2 cells. The dual luciferase reporter gene experiment was used to verify the targeted binding relationship between miR-4695-5p and Ras-related C3 botulinum toxin substrate 1 (RAC1). qRT-PCR and Western blot were used to detect the effect of overexpression of miR-4695-5p on the expression of RAC1 gene and Wnt/β-Catenin signaling pathway protein.The software of SPSS28.0 was used to conduct data analysis. The measurement data of normal distribution were espressed by Mean±SD. The t-test was used to compare the means between two groups, and the one-way analysis of variance was used to compare the means of multiple groups. Results:The expression level of miR-4695-5p in the serum of patients with colorectal cancer was significantly lower than that of healthy individuals ( P<0.01). The relative expression levels of miR-4695-5p in the control group and miR-4695-5p group were 1.09 ± 0.65 and 8.83±2.03, respectively. The expression level of miR-4695-5p in CACO-2 cells in the miR-4695-5p group was 8.10 times that of the control group, and CACO-2 cells overexpressing miR-4695-5p were successfully constructed ( P<0.01). Compared with the control group, the proliferation ability of CACO-2 cells in the miR-4695-5p group was significantly reduced ( P<0.05), and the invasion ability of CACO-2 cells was significantly reduced ( P<0.01). Bioinformatics tools and dual luciferase reporter gene experiments confirmed that miR-4695-5p can target and bind RAC1 ( P<0.01). Compared with the control group, the expression of RAC1 gene in the miR-4695-5p group was significantly decreased ( P<0.01), and the expression of Wnt/β-Catenin signaling pathway proteins Wnt3a, β-catenin, and c-MYC decreased significantly. Conclusions:miR-4695-5p is lowly expressed in the serum of colorectal cancer patients. miR-4695-5p inhibits the proliferation and invasion of colorectal cancer CACO-2 cells by targeting the inhibition of RAC1 gene expression and down-regulating the activity of the Wnt/β-Catenin signaling pathway.

Objective To investigate the effects of the number of harvested lymph nodes in neoadjuvant chemoradiotherapy (nCRT) combined with surgery on prognosis of middle-low rectal cancer.Methods The retrospective case-control study was conducted.The clinicopathological data of 373 patients with middle-low rectal cancer who underwent nCRT combined with surgery in the Fujian Medical University Union Hospital from January 2009 to December 2013 were collected.There were 241 males and 132 females,aged from 26 to 81 years,with the age of (55 ± 11) years.Observation indicators:(1) treatment situations;(2) follow-up and survival;(3)influencing factors for the number of harvested lymph nodes;(4) prognostic analysis of the different number of harvested lymph nodes as cut-off for grouping;(5) stratified analysis.Follow-up using telephone interview and outpatient examination was performed to detect postoperative survival of patients once every three months within postoperative 2 years and once every 6 months during the postoperative third year up to March 2016.The endpoint of follow-up was tumor recurrence,retastasis or death.Measurement data with normal distribution were represented as Mean±SD,and comparison between groups was done using the independent sample t test.Measurement data with skewed distribution were represented as M (range),and comparison between groups was done using the Kruska1-Wallis H test.Count data was described as absolute numbers.Univariate and multivariate analyses were done by the multiple linear regression model.Survival rate was calculated by the Kaplan-Meier method,and Logrank test was used for survival analysis.Results (l) Treatment situations:373 patients underwent nCRT combined with surgery,including 329 combined with sphincter-sparing rectal resection and 44 combined with abdominoperineal rectal resection.The number of harvested lymph nodes was 12 ± 6 in 373 patients.There were 185 patients with the number of harvested lymph nodes ＜ 12 and 188 with the number of harvested lymph nodes ≥ 12.(2) Follow-up and survival:373 patients were followed up for 5-77 months,with a median follow-up time of 43 months.During the follow-up,the 1-,3-,5-year disease-free survival rates were respectively 90.4％,76.3％,and 67.5％ in the 373 patients.(3) Influencing factors for the number of harvested lymph nodes:univariate analysis showed that distance between the tumor and anal verge,tumor diameter,tumor pathological N staging,and regression grade of rectal cancer were associated factors for the number of harvested lymph nodes (t =3.156,2.992,x2=8.183,10.839,P＜0.05).Multivariate analysis showed that distance between the tumor and anal verge,regression grade of rectal cancer,and tumor pathological N staging were independent factors for the number of harvested lymph nodes (t=3.308,2.690,2.584,95％ confidence interval:0.808-3.180,0.446-2.873,0.332-2.448,P＜0.05).(4) Prognostic analysis of the different number of harvested lymph nodes as cut-off for grouping:with the number of harvested lymph nodes of 6,7,8,9,10,11,12,13,14,15,and 16 as cut-off for grouping,there was no significant difference in the 3-year disease-free survival rate,cumulative local recurrence rate,and cumulative distant metastasis rate between ＜6 group and ≥6 group,between ＜7 group and ≥7 group,between＜8 group and ≥8 group,between ＜9 group and ≥9 group,between ＜10 group and ≥ 10 group,between ＜11 group and ≥ll group,between ＜12 group and ≥12 group,between ＜13 group and ≥13 group,between ＜ 14 group and ≥ 14 group,between ＜ 15 group and ≥ 15 group,between ＜ 16 group and ≥ 16 group,respectively (P＞0.05).(5) Stratified analysis:with the number of harvested lymph nodes of 7,8,9,and 10 as cut-off for grouping in 45 of 373 patients with Ⅱ-Ⅲ regression grade of rectal cancer and negative lymph nodes (NO staging),there was no significant difference in the 3-year disease-free survival rate between ＜7 group and ≥ 7group,between ＜8 group and ≥8 group,between ＜9 group and ≥9 group,between＜10 group and ≥ 10 group,respetively (x2 =3.946,5.346,6.375,4.297,P＜0.05).Conclusions The number of lymph nodes as 12 is not the independent factor for prognosis of patients with middle-low rectal cancer after nCRT combined with surgery.The number of harvested lymph nodes as 7 to 10 is the important factor for evaluating the prognosis of middle-low rectal cancer patients with Ⅱ-Ⅲ regression grade of rectal cancer and negative lymph nodes after nCRT combined with surgery.

OBJECTIVETo establish a nomogram to predict long-term survival in non-metastatic colorectal cancer patients.

METHODSA retrospective analysis was conducted in patients with non-metastatic colorectal cancer who underwent radical surgery in the Department of Colorectal Surgery of Affiliated Union Hospital of Fujian Medical University between January 2000 and December 2014. Univariate and multivariate analyses on disease-free survival (DFS) were performed using the Cox proportional regression model. Based on the multivariate analysis results, a prognostic nomogram was formulated to predict the probability for DFS. Concordance index was applied in predictive evaluation of the nomogram and calibration curves were drawn to test the nomogram's prediction and actual observation of the 5-year DFS rate. The predictive ability of nomogram was compared with AJCC-7 staging system.

RESULTSA total of 2 641 patients were identified. The median age was 59.3 years old, and 60.3% of cases were men. The number of patients with TNM stage 0, I(, II( and III( was 96, 505, 923 and 1043, respectively. The most common tumor site was the rectum, accounting for 43.2%. A total of 413 (15.6%) patients underwent neoadjuvant treatment. The most common gross type of tumor was ulcerative type, accounting for 79.5%. The 3- and 5-year DFS rate was 85.8% and 79.8%, respectively. Based on the Cox proportional regression model, the following six factors were independently associated with reduced DFS rate and were selected for the nomogram: older age, higher pathologic T stage, higher pathologic N stage, higher preoperative serum CEA level, infiltrative gross type and perineural invasion. The results of the nomogram showed that the score of T0, T1, T2, T3 and T4 stage was 0, 2.2, 3.9, 4.1 and 6, respectively, and the score of N0, N1 and N2 was 0, 3.8 and 9.3, respectively. For gross type, the score of expanding type, ulcerative type and infiltrative type was 6, 9 and 10, respectively. The score of perineural invasion was 5.2. Higher scores were added to older age and higher CEA level. The total scores were calculated by taking the sum of the points from all predictors. Higher total score was associated with poor DFS. The prognostic nomogram differentiated well and showed a concordance index of 0.718, which was better than AJCC-7 staging system (concordance index=0.683). Also, the calibration of nomogram predictions was good.

CONCLUSIONSA nomogram based on 6 independently prognostic factors to predict long-term survival in non-metastatic colorectal cancer patients is established successfully. The nomogram can be conveniently used to facilitate the accurate individualized prediction of DFS rates in patients with non-metastatic colorectal cancer.

OBJECTIVETo compare the short-term efficacy of laparoscopic and open transabdominal intersphincteric resection (ISR) for low rectal cancer.

METHODSClinicopathological data of 246 patients with low rectal cancer undergoing transabdominal ISR in our department from January 2005 to January 2015 were retrospectively analyzed. According to gender, age, ASA score, neoadjuvant chemoradiotherapy or not, pathological T stage, pathologic N stage, and tumor differentiation, propensity score matching was performed by R plug-in(version 2.8.1). Finally, 74 cases treated by laparoscopic transabdominal ISR(laparoscopic group) and 74 cases by open transabdominal ISR(open group) were enrolled. Short-term efficacy and anal function were compared between two groups.

RESULTSNo perioperative death was found in the two groups. Compared to open group, laparoscopic group had longer operation time [(236±45) minutes vs. (200±46) minutes, P=0.000], less median blood loss [50(10 to 200) ml vs. 100(20 to 400) ml, P=0.000] and shorter hospital stay [(7.8±2.4) days vs. (10.5±6.9) days, P=0.002]. Laparoscopic group and open group had similar morbidity of total complication [17.6%(13/74) vs. 28.4%(21/74), P=0.118]. Incidence of pneumonia was significantly lower in laparoscopic group [4.1%(3/74) vs. 13.5%(10/74), P=0.042), while incidence of anastomotic leakage and stenosis, and complication grading were not significantly different between the two groups (all P>0.05). During a mean follow-up of 52.0 months, anal function analysis was performed in 102 patients with stoma closure and the result showed that the ratio of patients with good continence was 87.1%(54/62) and 87.5%(35/40) in laparoscopic and open group respectively (P=0.066).

CONCLUSIONLaparoscopic transabdominal ISR is safe and feasible, which is minimally invasive with fast recovery, and is worth clinical application.

Aged ; Anastomotic Leak ; Chemoradiotherapy ; Female ; Humans ; Laparoscopy ; methods ; Length of Stay ; Male ; Middle Aged ; Neoadjuvant Therapy ; Operative Time ; Rectal Neoplasms ; surgery ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the influence of anastomotic leakage (AL) on long-term survival after resection for rectal cancer.

METHODSClinicopathological data of 653 rectal cancer cases confirmed by pathology and undergoing R0 resection for rectal cancer in our department from January 2007 to December 2011 were retrospectively analyzed. Anastomotic leakage was found in 40 cases (AL group) and not in the other 613 cases (non-AL group). After median 47 (1-91) months of follow-up, 5-year disease-free survival rate, distant metastasis rate and local recurrence rate were compared between the two groups. Risk factors affecting long-term prognosis were also analyzed.

RESULTSThe 5-year disease-free survival rate, 5-year distant metastasis rate, and 5-year local recurrence rate were 78.1%, 14.2% and 4.2% in the non-AL group, and 74.5%, 20.1% and 8.4% in the AL group respectively, and the differences were not statistically significant (P=0.808, P=0.965, P=0.309). Multivariate analysis showed that preoperative neoadjuvant radiochemotherapy, TNM staging, abnormal CA199, preoperative low level of albumin were independent prognostic factors of rectal cancer patients after R0 resection, while AL was not an independent factor of 5-year disease-free survival (P=0.910). Further multivariate analysis on 507 cases receiving postoperative adjuvant chemotherapy also revealed that AL was not an independent factor of 5-year disease-free survival (P>0.05). Percentage difference of patients finishing postoperative chemotherapy between the two groups was not statistically significant (79.4% vs. 76.3%, P=0.681).

CONCLUSIONAL is not an independent predictor of long-term survival for rectal cancer.

Anastomotic Leak ; Chemotherapy, Adjuvant ; Disease-Free Survival ; Humans ; Neoadjuvant Therapy ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Postoperative Period ; Prognosis ; Rectal Neoplasms ; pathology ; Retrospective Studies ; Survival Rate