1.Recurrence outcomes of robotic-versus laparoscopic-assisted gastrectomy for gastric cancer: a multi-center propensity score-matched cohort study
Jun LU ; Taiyuan LI ; Li ZHANG ; Junjun SHE ; Junyu CHEN ; Qing ZHONG ; Zukai WANG ; Changming HUANG ; Chaohui ZHENG
Chinese Journal of Gastrointestinal Surgery 2024;27(8):799-807
Objective:To compare and evaluate recurrence patterns after robotic-assisted gastrectomy (RAG) versus laparoscopic-assisted gastrectomy (LAG).Methods:This was a retrospective cohort study of 2915 consecutive patients with gastric adenocarcinoma confirmed by postoperative histology as T1-4aN0-3M0, who had undergone minimally invasive radical gastrectomy at four large gastric cancer treatment centers (Fujian Medical University Union Hospital: 1426 patients; the First Affiliated Hospital, Nanchang University: 1108; Tianjin Medical University Cancer Institute and Hospital: 196; and First Affiliated Hospital of Xi'an Jiaotong University: 185 cases) between 1 January 2015 and 30 June 2019. 930 patients had undergone RAG (RAG group) and 1985 had undergone LAG (LAG group). We assessed the following characteristics: age, sex, body mass index, American Society of Anesthesiologists score, comorbidities, tumor size, extent of surgery, extent of lymph node dissection, pT, pN, year of surgery, and adjuvant chemotherapy, after propensity score matching (1:1). There were no significant differences in baseline clinical characteristics between the two groups formed by propensity score matching (837 in each group) (all P>0.05). The 3-year recurrence-free survival (RFS), recurrence pattern, and conditional RFS were compared. Results:We detected no significant differences in the overall recurrence rate at 3 years (128/837 [15.3%] vs. 141/837 [16.8%], P=0.387) or time to recurrence (15.7±8.1 months vs. 16.4±8.4 months, P=0.449) between the RAG and LAG groups. Peritoneal recurrence was the most common type of recurrence in both groups (55 [6.6%] vs. 69 [8.2%], P=0.524). The difference in 3-year RFS between the RAG and LAG groups was not statistically significant (83.2% vs. 82.5%, P=0.781). We found that age > 60 years, total gastrectomy, and worse pT stage and pN stage were independent risk factors for recurrence in the study patients (all P<0.05), whereas the surgical procedure (RAG or LAG) was not an independent risk factor for RFS ( P=0.242). The 3-year conditional RFS at various time points was comparable between the two groups (1 year postoperatively: 84.6% vs. 84.7%, P=0.793; 3 years postoperatively: 91.5% vs. 94.9%, P=0.647). Conclusions:In this multicenter study of patients with locally resectable gastric cancer, we demonstrated that RAG performed by surgeons at large gastric cancer centers is not inferior to LAG in 3-year recurrence rate or recurrence patterns.
2.A real-world retrospective study of pelvic floor muscle training efficacy using an APP-based home device
Ji-Qiong ZHENG ; Shi-Ping ZHOU ; Zhong-Yun ZHANG ; Wei-Qing QIAN ; Zheng-Wang ZHANG
Fudan University Journal of Medical Sciences 2024;51(1):96-101
Objective To retrospectively investigate the efficacy of pelvic floor muscle training(PFMT)using APP-based home device in the real-world scenario.Methods A retrospective survey was conducted among 171 000 Chinese female users over 18 years old who freely registered APP and performed PFMT using a domestic manufactured home device from Oct 1,2019 to Mar 31,2021.The Patient Global Impression of Improvement(PGI-I)questionnaires concerning pelvic floor general condition,sexual satisfaction and other three major symptoms related to pelvic floor muscle laxity such as stress urinary incontinence(SUI),pelvic organ prolapse(POP),and vaginal laxity(VL)were broadcast public online for users to self-assess the improvement.The vaginal muscle strength values prior and post training recorded by APP were also reported.All data were collected anonymously for further stratified analysis.Results A total of 984 valid questionnaires were collected by systematic sampling.The PGI-I scores of pelvic floor general condition(P=0.000 1),sexual satisfaction(P=0.009),SUI(P=0.000 1),POP(P=0.044)and VL(P=0.034)were statistically significant in users who reported to use the device for 3 months or more compared with those less than 3 months.In addition,the increase of vaginal muscle strength was related to the improvement of PGI-I scores in SUI and POP with statistical significance.There were no significant difference in subgroups such as age,education,parity,and delivery mode.Conclusion The subjective benefit of pelvic floor function-related symptoms and the improvement of vaginal muscle strength could be observed after PFMT for three months or more using APP-based home device in Chinese women in the real-world scenario.
3.Establishment of a rapid detection method for Pseudomonas aeruginosa based on real-time fluorescent recombinase polymerase amplification technology
Yun XING ; Yan ZHANG ; Daohong ZHOU ; Qiu ZHONG ; Yuansu JIANG ; Qing HUANG ; Baihui ZHENG
International Journal of Laboratory Medicine 2024;45(19):2329-2333
Objective To establish a real-time fluorescent recombinase polymerase amplification(RPA)technology for the detection of the virulence gene exoS of Pseudomonas aeruginosa,and evaluate the specifici-ty,sensitivity and practicability of the method.Methods According to the specific conserved region of the vir-ulence gene exoS of Pseudomonas aeruginosa,the specific primers and probes of RPA were designed,and the extracted target DNA was detected to determine the specificity and sensitivity of RPA.Real-time fluorescence quantitative PCR(qPCR)was established to detect the target DNA,and the detection limits of different detec-tion methods for Pseudomonas aeruginosa were compared.The feasibility of RPA in detecting Pseudomonas aeruginosa was further confirmed by the performance verification test of clinical samples.Results The estab-lished RPA detection method had good specificity.Only Pseudomonas aeruginosa had specific amplification curve,but no specific amplification curve for other bacteria.The sensitivity of RPA was 5×102 cfu/mL,which was consistent with the detection limit of qPCR and the results were reliable.The detection time of RPA method was only 30 min,which was significantly lower than that of the traditional method.Conclusion The RPA method for the detection of Pseudomonas aeruginosa established in this study has high specificity and sensitivity,and significantly shortens the detection time compared with the traditional detection method.It can be used for the rapid detection of Pseudomonas aeruginosa in clinical specimens.
4.Recurrence outcomes of robotic-versus laparoscopic-assisted gastrectomy for gastric cancer: a multi-center propensity score-matched cohort study
Jun LU ; Taiyuan LI ; Li ZHANG ; Junjun SHE ; Junyu CHEN ; Qing ZHONG ; Zukai WANG ; Changming HUANG ; Chaohui ZHENG
Chinese Journal of Gastrointestinal Surgery 2024;27(8):799-807
Objective:To compare and evaluate recurrence patterns after robotic-assisted gastrectomy (RAG) versus laparoscopic-assisted gastrectomy (LAG).Methods:This was a retrospective cohort study of 2915 consecutive patients with gastric adenocarcinoma confirmed by postoperative histology as T1-4aN0-3M0, who had undergone minimally invasive radical gastrectomy at four large gastric cancer treatment centers (Fujian Medical University Union Hospital: 1426 patients; the First Affiliated Hospital, Nanchang University: 1108; Tianjin Medical University Cancer Institute and Hospital: 196; and First Affiliated Hospital of Xi'an Jiaotong University: 185 cases) between 1 January 2015 and 30 June 2019. 930 patients had undergone RAG (RAG group) and 1985 had undergone LAG (LAG group). We assessed the following characteristics: age, sex, body mass index, American Society of Anesthesiologists score, comorbidities, tumor size, extent of surgery, extent of lymph node dissection, pT, pN, year of surgery, and adjuvant chemotherapy, after propensity score matching (1:1). There were no significant differences in baseline clinical characteristics between the two groups formed by propensity score matching (837 in each group) (all P>0.05). The 3-year recurrence-free survival (RFS), recurrence pattern, and conditional RFS were compared. Results:We detected no significant differences in the overall recurrence rate at 3 years (128/837 [15.3%] vs. 141/837 [16.8%], P=0.387) or time to recurrence (15.7±8.1 months vs. 16.4±8.4 months, P=0.449) between the RAG and LAG groups. Peritoneal recurrence was the most common type of recurrence in both groups (55 [6.6%] vs. 69 [8.2%], P=0.524). The difference in 3-year RFS between the RAG and LAG groups was not statistically significant (83.2% vs. 82.5%, P=0.781). We found that age > 60 years, total gastrectomy, and worse pT stage and pN stage were independent risk factors for recurrence in the study patients (all P<0.05), whereas the surgical procedure (RAG or LAG) was not an independent risk factor for RFS ( P=0.242). The 3-year conditional RFS at various time points was comparable between the two groups (1 year postoperatively: 84.6% vs. 84.7%, P=0.793; 3 years postoperatively: 91.5% vs. 94.9%, P=0.647). Conclusions:In this multicenter study of patients with locally resectable gastric cancer, we demonstrated that RAG performed by surgeons at large gastric cancer centers is not inferior to LAG in 3-year recurrence rate or recurrence patterns.
5.Application and practice of indocyanine green-guided laparoscopic radical gastrec-tomy for gastric cancer
Huang CHANGMING ; Liang HAN ; Zheng CHAOHUI ; Chen QIYUE ; Zhong QING ; Qiu TAOYUAN
Chinese Journal of Clinical Oncology 2024;51(13):663-669
Indocyanine green(ICG)near-infrared imaging technology has significant research value in laparoscopic radical gastrectomy and has garnered widespread attention and research both domestically and internationally.However,the application of ICG near-infrared ima-ging technology in laparoscopic radical gastrectomy is still in the exploratory stage in current clinical practice,with no unified standards es-tablished as yet.This review introduces the mechanism of ICG fluorescence imaging and discusses the indications and contraindications of ICG use in laparoscopic radical gastrectomy.The application methods and procedures of ICG molecular fluorescence imaging technology in laparoscopic radical gastrectomy and its clinical applications are elaborated upon.Finally,the practical applications of ICG-guided laparo-scopic radical gastrectomy is summarized and generalized to help facilitate its promotion and further standardization.
6.A multi-center epidemiological study on pneumococcal meningitis in children from 2019 to 2020
Cai-Yun WANG ; Hong-Mei XU ; Gang LIU ; Jing LIU ; Hui YU ; Bi-Quan CHEN ; Guo ZHENG ; Min SHU ; Li-Jun DU ; Zhi-Wei XU ; Li-Su HUANG ; Hai-Bo LI ; Dong WANG ; Song-Ting BAI ; Qing-Wen SHAN ; Chun-Hui ZHU ; Jian-Mei TIAN ; Jian-Hua HAO ; Ai-Wei LIN ; Dao-Jiong LIN ; Jin-Zhun WU ; Xin-Hua ZHANG ; Qing CAO ; Zhong-Bin TAO ; Yuan CHEN ; Guo-Long ZHU ; Ping XUE ; Zheng-Zhen TANG ; Xue-Wen SU ; Zheng-Hai QU ; Shi-Yong ZHAO ; Lin PANG ; Hui-Ling DENG ; Sai-Nan SHU ; Ying-Hu CHEN
Chinese Journal of Contemporary Pediatrics 2024;26(2):131-138
Objective To investigate the clinical characteristics and prognosis of pneumococcal meningitis(PM),and drug sensitivity of Streptococcus pneumoniae(SP)isolates in Chinese children.Methods A retrospective analysis was conducted on clinical information,laboratory data,and microbiological data of 160 hospitalized children under 15 years old with PM from January 2019 to December 2020 in 33 tertiary hospitals across the country.Results Among the 160 children with PM,there were 103 males and 57 females.The age ranged from 15 days to 15 years,with 109 cases(68.1% )aged 3 months to under 3 years.SP strains were isolated from 95 cases(59.4% )in cerebrospinal fluid cultures and from 57 cases(35.6% )in blood cultures.The positive rates of SP detection by cerebrospinal fluid metagenomic next-generation sequencing and cerebrospinal fluid SP antigen testing were 40% (35/87)and 27% (21/78),respectively.Fifty-five cases(34.4% )had one or more risk factors for purulent meningitis,113 cases(70.6% )had one or more extra-cranial infectious foci,and 18 cases(11.3% )had underlying diseases.The most common clinical symptoms were fever(147 cases,91.9% ),followed by lethargy(98 cases,61.3% )and vomiting(61 cases,38.1% ).Sixty-nine cases(43.1% )experienced intracranial complications during hospitalization,with subdural effusion and/or empyema being the most common complication[43 cases(26.9% )],followed by hydrocephalus in 24 cases(15.0% ),brain abscess in 23 cases(14.4% ),and cerebral hemorrhage in 8 cases(5.0% ).Subdural effusion and/or empyema and hydrocephalus mainly occurred in children under 1 year old,with rates of 91% (39/43)and 83% (20/24),respectively.SP strains exhibited complete sensitivity to vancomycin(100% ,75/75),linezolid(100% ,56/56),and meropenem(100% ,6/6).High sensitivity rates were also observed for levofloxacin(81% ,22/27),moxifloxacin(82% ,14/17),rifampicin(96% ,25/26),and chloramphenicol(91% ,21/23).However,low sensitivity rates were found for penicillin(16% ,11/68)and clindamycin(6% ,1/17),and SP strains were completely resistant to erythromycin(100% ,31/31).The rates of discharge with cure and improvement were 22.5% (36/160)and 66.2% (106/160),respectively,while 18 cases(11.3% )had adverse outcomes.Conclusions Pediatric PM is more common in children aged 3 months to under 3 years.Intracranial complications are more frequently observed in children under 1 year old.Fever is the most common clinical manifestation of PM,and subdural effusion/emphysema and hydrocephalus are the most frequent complications.Non-culture detection methods for cerebrospinal fluid can improve pathogen detection rates.Adverse outcomes can be noted in more than 10% of PM cases.SP strains are high sensitivity to vancomycin,linezolid,meropenem,levofloxacin,moxifloxacin,rifampicin,and chloramphenicol.[Chinese Journal of Contemporary Pediatrics,2024,26(2):131-138]
7.Correlation analysis and benchmark dose study on bone metabolic biochemical index of low doses of exposed hydrogen fluoride workers.
Guang ZHENG ; Feng Yi LI ; Xiang WANG ; Dong Qing ZHU ; Zhong Lin ZHAO ; Ying GUO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2023;41(3):198-203
Objective: To analyze correlation of occupational hydrogen fluoride exposure to low doses of bone metabolism index through occupational epidemiological investigation and benchmark dose calculation. Methods: In May 2021, using cluster sampling method, 237 workers exposed to hydrogen fluoride in a company were selected as the contact group, and 83 workers not exposed to hydrogen fluoride in an electronics production company were selected as the control group. The external exposure dose and urinary fluoride concentration, blood and urine biochemical indicators of the workers was measured.The relationship between external dose and internal dose of hydrogen fluoride was analyzed. The external dose, urinary fluoride was used as exposure biomarkers, while serum osteocalcin (BGP), serum alkaline phosphatase (AKP) and urinary hydroxyproline (HYP) were used as effect biomarkers for bone metabolism of hydrogen fluoride exposure. The benchmark dose calculation software (BMDS1.3.2) was used to calculate benchmark dose (BMD) . Results: Urine fluoride concentration in the contact group was correlated with creatinine-adjusted urine fluoride concentration (r=0.69, P=0.001). There was no significant correlation between the external dose of hydrogen fluoride and urine fluoride in the contact group (r=0.03, P=0.132). The concentrations of urine fluoride in the contact group and the control group were (0.81±0.61) and (0.45±0.14) mg/L, respectively, and the difference between the two groups was statistically significant (t=5.01, P=0.025). Using BGP, AKP and HYP as effect indexes, the urinary BMDL-05 values were 1.28, 1.47 and 1.08 mg/L, respectively. Conclusion: Urinary fluoride can sensitively reflect the changes in the effect indexes of biochemical indexes of bone metabolism. BGP and HYP can be used as early sensitive effect indexes of occupational hydrogen fluoride exposure.
Humans
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Fluorides/adverse effects*
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Hydrofluoric Acid
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Benchmarking
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Biomarkers
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Occupational Exposure/adverse effects*
8.Establishment of a nomogram prediction model using common preoperative indicators for early weight loss after laparoscopic sleeve gastrectomy
Yiming JIANG ; Jing JIA ; Qing ZHONG ; Qiyue CHEN ; Jun LU ; Jiabin WANG ; Jianwei XIE ; Ping LI ; Zhaohui ZHENG ; Changming HUANG ; Xinyu LI ; Jianxian LIN
Chinese Journal of Gastrointestinal Surgery 2023;26(11):1058-1063
Objectives:To construct a nomogram prediction model using common preoperative indicators for early weight loss (EWL) 1 year after laparoscopic sleeve gastrectomy (LSG).Methods:Relevant data of obese patients who had undergone LSG from January 2015 to May 2022 in Fujian Medical University Union Hospital and Quanzhou First Hospital Affiliated Fujian Medical University were analyzed. Patients with a history of major abdominal surgery, severe gastroesophageal reflux disease, pregnancy within 1 year after surgery, or who were lost to follow-up were excluded, resulting in a total of 200 patients in the study (190 from Fujian Medical University Union Hospital and 10 from Quanzhou First Hospital Affiliated Fujian Medical University). The participants were 51 men and 149 women of a mean age 29.9±8.2 years and a body mass index (BMI) 38.7±6.5 kg/m 2. All patients in this group underwent standardized LSG procedure. Achieving ideal weight (BMI≤25 kg/m 2) 1 year after LSG was defined as goal of EWL. Logistic regression analyses were performed to identify factors that independently influenced EWL. These factors were incorporated into the nomogram model. Receiver operating characteristic (ROC) curves (the larger the area under the curve [AUC], the better the predictive ability and accuracy of the model), likelihood ratio test (higher likelihood ratio indicates greater model homogeneity), decision curve analysis (higher net benefit indicates a better model), Akaike information criterion (AIC; smaller AIC indicates a better model), and Bayesian information criterion (BIC; smaller BIC indicates a better model) were used to validate the predictive ability of the column line diagram model. Results:In this study of 200 obese patients who underwent LSG surgery, 136 achieved EWL goal, whereas the remaining 64 did not. The rate of EWL goal achievement of the entire group was 68.0%. Compared with patients who did not achieve EWL goal, those who did had lower BMI, alanine transaminase, aspartate transaminase, triglycerides, and higher cholesterol. Additionally, the proportion of female was higher and the proportions of patients with fatty liver and hypertension lower in those who achieved EWL goal (all P<0.05). Univariate and multivariate logistic regression analysis revealed that preoperative BMI (OR=0.852, 95%CI: 0.796-0.912, P<0.001), alanine transaminase (OR=0.992, 95%CI: 0.985-0.999, P=0.024), presence of fatty liver (OR=0.185, 95%CI: 0.038-0.887, P=0.035) and hypertension (OR=0.374, 95%CI: 0.144-0.969, P=0.043) were independently associated with failure to achieve EWL goal. Cholesterol (OR=1.428, 95%CI: 1.052-1.939, P=0.022) was independently associated with achieving EWL goal. We used the above variables to establish an EWL nomogram model. ROC analysis, the likelihood ratio test, decision curve analysis, and AIC all revealed that the predictive value of the model was better than that of BMI alone (nomogram model vs. BMI: area under the curve 0.840 vs. 0.798, P=0.047; likelihood ratio: 58.785 vs. 36.565, AIC: 193.066 vs. 207.063, BIC: 212.856 vs. 213.660). Conclusion:Our predictive model is more accurate in predicting EWL after LSG compared with using BMI.
9.Learning curve analysis and influencing factors of operation time of laparoscopic sleeve gastrectomy
Zhixin SHANGGUAN ; Qing ZHONG ; Yiming JIANG ; Chaohui ZHENG ; Ping LI ; Jianwei XIE ; Jiabin WANG ; Jun LU ; Jianxian LIN ; Changming HUANG
Chinese Journal of Digestive Surgery 2023;22(8):996-1002
Objective:To investigate the influencing factors of operation time for laparos-copic sleeve gastrectomy (LSG) and analyze the learning curve of LSG in sarcopenic obesity (SO) and non-sarcopenic obesity (NSO).Methods:The retrospective cohort study was conducted. The clinical data of 240 obesity patients who underwent LSG in the Fujian Medical University Union Hospital from January 2018 to June 2022 were collected. There were 52 males and 188 females, aged (30±8)years. Patients underwent L3 vertebral body horizontal axial computer tomography (CT) scanning before and after receiving LSG to accurately segment muscles and fats. Observation indicators: (1) treatment and follow-up; (2) influencing factors of operation time for LSG; (3) cumulative sum (CUSUM) of learning curve; (4) comparison of clinical data between patients in the initial and profi-cient stages. Measurement data with normal distribution were represent as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M(IQR), and comparison between groups was conducted using the non-parameter test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test. Univariate and multivariate analyses were conducted using the Logistic regression model. The CUSUM of learning curve was calculated and the fitting process was conducted on scatter plot of learning curves. Results:(1) Treatment and follow-up. Of the 240 patients, there were 97 cases of SO and 143 cases of NSO. All 240 patients underwent LSG successfully, without conversion to open surgery. The operation time of 240 patients was (108±23)minutes. None of patient died during the perioperative period and all patients underwent follow-up during the postoperative 6 months. (2) Influencing factors of operation time for LSG. Results of multivariate analysis showed that SO was an independent factor influencing operation time for LSG ( odds ratio=2.207, 95% confidence interval as 1.207-4.038, P<0.05). (3) CUSUM of learning curve. Results of CUSUM of operation time in patients of SO and NSO showed that the best fit equation of patients of SO was y=-4E-08x 6+1E-05x 5-0.001 1x 4+0.063 1x 3-1.89x 2+28.126x-48.671 (x means the number of surgical cases), with goodness-of-fit R 2 as 0.833, and the best fit equation of patients of NSO was y=3E-09x 6-1E-06x 5+0.000 2x 4-0.010 9x 3+0.063 8x 2+12.053x-65.025 (x means the number of surgical cases), with goodness-of-fit R 2 as 0.716. Based on the trend of CUSUM of learning curve of operation time, the peak value of number of surgical cases in patients of SO and NSO was 81 and 36, respec-tively, which was used to divide the learning curve as two stages of the initial stage and the proficient stage. (4) Comparison of clinical data between patients in the initial and proficient stages. ① Of the 97 patients of SO, there were 81 cases and 16 cases in the initial stage and the proficient stage of LSG, with the operation time, postoperative duration of hospital stay as (119±23)minutes, (5.9±2.3)days and (106±21)minutes, (4.7±0.5)days, showing significant differences between them ( t=2.074, 2.147, P<0.05). ②Of the 143 patients of NSO, there were 36 cases and 107 cases in the initial stage and the proficient stage of LSG, with gender (female), height, preoperative body mass, defatted body mass, operation time, postoperative duration of hospital stay, body mass at postoperative 6 month, body mass index (BMI) at postoperative 6 month, percentage of excess weight loss (EWL%) at postoperative 6 month, cases with EWL% >100% at postoperative 6 month, excess BMI at post-operative 6 month as 20, (170±10)cm, (110±25)kg, (57±12)kg, (108±22)minutes, (6.1±1.6)days, (80±16)kg, (27.63±4.22)kg/m2, 83%±35%, 9, 1.99(6.03)kg/m2 and 87, (164±8)cm, (99±20)kg, (52±12)kg, (100±19)minutes, (4.7±1.1)days, (71±16)kg, (25.89±4.48)kg/m2, 103%±42%, 48, 0.31(5.82)kg/m2, showing significant differences between them ( χ2=9.484, t=3.266, 2.424, 2.141, 2.137, 5.821, 2.740, 1.993, -2.524, χ2=4.432, Z=-2.300, P<0.05). Conclusions:SO is an independent factor influencing operation time for LSG. It is suggested that the surgeons need to finish 81 cases and 36 cases master LSG in patients of SO and NSO.
10.The mechanism of action and prognostic value of Dynamin 3 in gastric cancer
Ruhong TU ; Gildas Eric Sita Emmanuel ; Qing ZHONG ; Chaohui ZHENG ; Ping LI ; Jianwei XIE ; Jiabin WANG ; Jianxian LIN ; Jun LU ; Qiyue CHEN ; Longlong CAO ; Mi LIN ; Changming HUANG
Chinese Journal of Digestive Surgery 2023;22(9):1100-1112
Objective:To investigate the mechanism of action and prognostic value of Dynamin 3 (DNM3) in gastric cancer.Methods:The bioinformatic analysis, experimental study and retrospective cohort study was conducted. The clinicopathological data, fresh gastric cancer tissues, paired normal tissues and the corresponding paraffin sections of 153 gastric cancer patients who underwent radical gastrectomy in Fujian Medical University Union Hospital from January 2013 to July 2018 were collected. Tissues and the corresponding paraffin sections were subjected to quanti-tative real-time polymerase chain reaction, immunoblotting assay, flow cytometric cell cycle assay and immunohistochemical staining, respectively, and clinicopathological data were used for prognostic analysis. The stomach adenocarcinoma (STAD) dataset from the Cancer Genome Atlas (TCGA) database was collected for bioinformatic analysis. Observation indicators: (1) DNM3 gene expression in TCGA-STAD in gastric cancer; (2) mutations and copy number alterations of DNM3 in gastric cancer; (3) methylation level of promoter of DNM3 in gastric cancer; (4) relative protein expression of DNM3 and p53 in gastric cancer; (5) DNM3 correlation and enrichment analysis; (6) ratio of G0/G1 phase, S phase and G2/M phase of cell cycle progression; (7) correlation between immune cell infiltration and DNM3 in gastric cancer; (8) correlation between results of immunohistochemical (IHC) staining and clinical features; (9) analysis of independent factors influencing 5-year overall survival rate of gastric cancer patients. Measurement data with normal distribution were represented as Mean±SD, and comparison among multiple groups was conducted using the ANOVA and further comparison between two groups was conducted using the LSD. Comparison between two groups was conducted using the t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and compari-son between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the rank sum test. The Pearson correlation coefficient or Spearman correlation coefficient was used to test the correlation between groups. Univariate and multivariate analyses were conducted using the COX proportional risk regression model. The Kaplan-Meier method was used to draw survival curves and calculate survival rates, and the Log-Rank test was used for survival analysis. The Benjamini-Hochberg false discovery rate correction was used for adjusting of the P-value. Results:(1) DNM3 gene expression in TCGA-STAD. The expression levels of DNM3 gene in the 27 tumor tissues and paired normal tissues of the TCGA-STAD database were 0.775(0.605,1.161) and 1.216(0.772,1.681), showing a significant difference between them ( Z=?2.64, P<0.05). The messenger RNA (mRNA) expression levels of DNM3 gene in 48 pairs of gastric cancer tissues and paired normal tissues of the author′s center were 4.370(2.870,6.040) and 2.520(0.850,4.170), showing a significant difference between them ( Z=?4.39, P<0.05). (2) Mutations and copy number alterations of DNM3 in gastric cancer. There were 16 gastric cancer patients in the TCGA-STAD database with DNM3 mutation or somatic copy number alterations, including 6 cases with missense mutations, 1 case with truncated mutation, 8 cases with copy number gain and 1 case with copy number loss. The mRNA expression levels of DNM3 gene before and after mutation in the 370 gastric cancer patients of the TCGA-STAD database were 6.13(5.40,7.08) and 5.02(3.98,5.46), showing a significant difference between them (Log 2FC=?1.11, Z=?2.59, P<0.05). (3) Methylation level of promoter of DNM3 in gastric cancer. There were 372 gastric cancer patients in the TCGA-STAD database undergoing DNM3 methylation and mRNA examinations, and the results showed that levels of methylation and mRNA expression of DNM3 was 0.198 (-0.458, 0.301) and 6.014 (5.141, 6.628), respectively. The levels of methylation in DNM3 was negatively correlated with its mRNA expression ( r=?0.38, P<0.05). Results of follow-up in 32 patients showed that the 3-year overall survival rate of 16 cases with high levels of methylation in DNM3 and 16 cases with low levels of methylation in DNM3 was 18.8% and 41.3%, respectively, showing a significant difference between them ( hazard ratio=1.40, P<0.05). Results of immunoblot-ting assay showed that the relative expression level of DNM3 protein in the AGS cells treated with 0, 0.5, and 1.0 μmol/L of 5-azacytidin was 0.270±0.020, 0.357±0.051 and 0.599±0.039, respectively, showing a significant difference among the three groups ( F=57.84, P<0.05). The relative expression level of DNM3 protein in the HGC-27 cells treated with 0, 0.5, and 1.0 μmol/L of 5-azacytidin was 0.316±0.038, 0.770±0.031 and 0.877±0.052, respectively, showing a significant difference among the three groups ( F=156.30, P<0.05). (4) Relative protein expression of DNM3 and p53 in gastric cancer. Results of immunoblotting assay showed that the relative expression of DNM3 and p53 protein was 0.688±0.047 and 0.872±0.041 in the AGS cells transfected with pCMV-DNM3 plasmid, versus 0.249±0.029 and 0.352±0.020 in the AGS cells transfected with control plasmid, showing significant differences in the above indicators between the two types of cells ( t=13.77,19.74, P<0.05). The relative expression of DNM3 and p53 protein was 0.969±0.069 and 1.464±0.081 in the HGC-27 cells transfected with pCMV-DNM3 plasmid, versus 0.456±0.048 and 0.794±0.052 in the HGC-27 cells transfected with control plasmid, showing significant differences in the above indicators between the two types of cells ( t=10.57, 12.06, P<0.05). (5) DNM3 correlation and enrichment analysis. Results of correlation analysis showed that DNM3 was positively correlated with genes such as RBMS3, CNTN4 and PDE1A ( r=0.52, 0.52, 0.50, P<0.05) and negatively correlated with genes such as SLC25A39, PAICS and GAPDH ( r=?0.41, ?0.40, ?0.40, P<0.05) in gastric cancer. Results of gene set enrichment analysis showed that the set of genes related to ribosome and oxidative phosphorylation were upregulated in gastric cancer patients with DNM3 low expression [normalized enrichment score (NES)=?3.30, ?2.16, P<0.05], while the set of genes related to immunomodulatory interactions between lymphocytes and non-lymphoid cells were upregulated in gastric cancer patients with DNM3 high expression (NES=1.67, P<0.05). Results of gene ontology analysis showed that the low expression of DNM3 was associated with the separation of mitotic sister chromatid (No.0000070), nonsense-mediation of nuclear transcriptional mRNA catabolic process, sister chromatid separation (No.0000819), nuclear transcriptional mRNA catabolic process and regulation of oxidative phos-phorylation (NES=?2.29, ?3.10, ?2.33, ?2.56, ?2.68, P<0.05). Results of Kyoto encycl opedia of genes and genomes analysis showed that metabolic pathway related to ribosome and oxidative phosphory-lation were upregulated and crosstalked in gastric cancer with low expression of DNM3 (NES=?3.34, ?2.21, P<0.05). (6) Ratio of G0/G1 phase, S phase and G2/M phase of cell cycle progression. Results of flow cytometric cell cycle experiments showed that the proportions of G0/G1 phase, S phase and G2/M phase in the cell cycle was 65.1%±3.0%, 17.3%±3.0% and 17.6%±1.0% in the AGS cells transfected with pCMV-DNM3 plasmid, versus 53.4%±4.0%, 26.3%±2.0% and 20.3%±3.0% in the AGS cells transfected with control plasmid, showing significant differences in the proportions of G0/G1 phase and S phase in the two types of cells ( t=4.05, 4.32, P<0.05). (7) Correlation between immune cell infiltration and DNM3 in gastric cancer. Results of immune cell infiltration examination showed that the expression level of DNM3 was positively associated with mast cells, NK cells, pDCs, B cells, follicular helper T cells, effector memory T cells, T cells, central memory T cells, CD8 T cells, DC cells, macrophages, γ-δ T cells (Tgd), iDCs and eosinophils infiltration (Spearman correlation coefficients as 0.41, 0.29, 0.26, 0.20, 0.22, 0.22, 0.13, 0.16, 0.15, 0.14, 0.14, 0.17, 0.18, 0.22, P<0.05) and negatively associated with Th17 cell, Th2 cells and NK CD56 dim cells infiltration ( r=?0.18, ?0.23, ?0.10, P<0.05). (8) Correlation between results of IHC staining and clinical features. Results of IHC staining analysis showed that the IHC score of DNM3 was 3(2,4) in the 105 gastric cancer tissues, versus 6(4,9) in the 105 paired normal tissues, showing a significant difference between them ( Z=-7.35, P<0.05). There were significant differences in gender, tumor location and N stating between the 70 patients with low expression of DNM3 and the 35 patients with high expression of DNM3 ( χ2=4.29, 7.67, 6.86, P<0.05). (9) Analysis of independent factors influencing 5-year overall survival rate of gastric cancer patients. Results of multivariate analysis showed that stage pT3?4 and low IHC score of DNM3 were independent risk factors for 5-year overall survival rate of gastric cancer patients ( hazard ratio=1.91, 0.51, 95% confidence interval as 1.06?3.43, 0.26?0.98, P<0.05). The 5-year overall survival rate was 44.3% in patients with low expression of DNM3, versus 65.7% in gastric cancer patients with high expression of DNM3, showing a significant difference between them ( χ2=5.02, P<0.05). Conclusion:DNM3 is a tumor suppressor and an independent predictor of poor prognosis for gastric cancer, which may regulate gastric cancer cell cycle and immunosuppression in the tumor microenvironment through methylation.

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