1.Analysis of The Characteristics of Brain Functional Activity in Gross Motor Tasks in Children With Autism Based on Functional Near-infrared Spectroscopy Technology
Wen-Hao ZONG ; Qi LIANG ; Shi-Yu YANG ; Feng-Jiao WANG ; Meng-Zhao WEI ; Hong LEI ; Gui-Jun DONG ; Ke-Feng LI
Progress in Biochemistry and Biophysics 2025;52(8):2146-2162
ObjectiveBased on functional near-infrared spectroscopy (fNIRS), we investigated the brain activity characteristics of gross motor tasks in children with autism spectrum disorder (ASD) and motor dysfunctions (MDs) to provide a theoretical basis for further understanding the mechanism of MDs in children with ASD and designing targeted intervention programs from a central perspective. MethodsAccording to the inclusion and exclusion criteria, 48 children with ASD accompanied by MDs were recruited into the ASD group and 40 children with typically developing (TD) into the TD group. The fNIRS device was used to collect the information of blood oxygen changes in the cortical motor-related brain regions during single-handed bag throwing and tiptoe walking, and the differences in brain activation and functional connectivity between the two groups of children were analyzed from the perspective of brain activation and functional connectivity. ResultsCompared to the TD group, in the object manipulative motor task (one-handed bag throwing), the ASD group showed significantly reduced activation in both left sensorimotor cortex (SMC) and right secondary visual cortex (V2) (P<0.05), whereas the right pre-motor and supplementary motor cortex (PMC&SMA) had significantly higher activation (P<0.01) and showed bilateral brain region activity; in terms of brain functional integration, there was a significant decrease in the strength of brain functional connectivity (P<0.05) and was mainly associated with dorsolateral prefrontal cortex (DLPFC) and V2. In the body stability motor task (tiptoe walking), the ASD group had significantly higher activation in motor-related brain regions such as the DLPFC, SMC, and PMC&SMA (P<0.05) and showed bilateral brain region activity; in terms of brain functional integration, the ASD group had lower strength of brain functional connectivity (P<0.05) and was mainly associated with PMC&SMA and V2. ConclusionChildren with ASD exhibit abnormal brain functional activity characteristics specific to different gross motor tasks in object manipulative and body stability, reflecting insufficient or excessive compensatory activation of local brain regions and impaired cross-regions integration, which may be a potential reason for the poorer gross motor performance of children with ASD, and meanwhile provides data support for further unraveling the mechanisms underlying the occurrence of MDs in the context of ASD and designing targeted intervention programs from a central perspective.
2.Clustering analysis of risk factors in high-incidence areas of esophageal cancer in Yanting county
Ruiwu LUO ; Heng HUANG ; Hao CHENG ; Siyu NI ; Siyi FU ; Qinchun QIAN ; Junjie YANG ; Xinlong CHEN ; Hanyu HUANG ; Zhengdong ZONG ; Yujuan ZHAO ; Yuhe QIN ; Chengcheng HE ; Ye WU ; Hongying WEN ; Dong TIAN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(03):385-391
Objective To investigate the dietary patterns of rural residents in the high-incidence areas of esophageal cancer (EC), and to explore the clustering and influencing factors of risk factors associated with high-incidence characteristics. Methods A special structured questionnaire was applied to conduct a face-to-face survey on the dietary patterns of rural residents in Yanting county of Sichuan Province from July to August 2021. Univariate and multivariate logistic regression models were used to analyze the influencing factors of risk factor clustering for EC. Results There were 838 valid questionnaires in this study. A total of 90.8% of rural residents used clean water such as tap water. In the past one year, the people who ate fruits and vegetables, soybean products, onions and garlic in high frequency accounted for 69.5%, 32.8% and 74.5%, respectively; the people who ate kimchi, pickled vegetables, sauerkraut, barbecue, hot food and mildew food in low frequency accounted for 59.2%, 79.6%, 68.2%, 90.3%, 80.9% and 90.3%, respectively. The clustering of risk factors for EC was found in 73.3% of residents, and the aggregation of two risk factors was the most common mode (28.2%), among which tumor history and preserved food was the main clustering pattern (4.6%). The logistic regression model revealed that the gender, age, marital status and occupation were independent influencing factors for the risk factors clustering of EC (P<0.05). Conclusion A majority of rural residents in high-incidence areas of EC in Yanting county have good eating habits, but the clustering of some risk factors is still at a high level. Gender, age, marital status, and occupation are influencing factors of the risk factors clustering of EC.
3.Applications of Three Dimensional Printing in Chromatography Separation
Qiang LI ; Hao-Nan LU ; Han-Rong WEN ; Zong-Peng WANG ; Bo ZHANG
Chinese Journal of Analytical Chemistry 2024;52(5):624-633
Chromatography is one of the most important separation and analytical techniques in production activity and academic research.With the growing demand for applications,the development of targeted separation devices is costly.However,the difficulty of constructing modeling makes it difficult to validate the theoretical studies of chromatography.Three dimensional(3D)printing,as a technology that can fabricate objects by depositing materials from the bottom to up,can custom print complex structures for specific needs,and shows many advantages such as low cost,low waste,high precision,high flexibility and parallel manufacturing,demonstrating great potential in the field of chromatography separations.In recent years,with the rapid development of 3D printing technology,the printing resolution and speed have progressively improved,and the range of printable materials has largely expanded.This has led to preliminary research and application of 3D printing technology in the field of chromatography separation,resulting in brand new discoveries and technological innovations.This article made a comprehensive overview of the latest advancements in research and application of 3D printing technology in separation science,including 3D printed columns,3D printed stationary phase,and 3D printed solid-phase extraction devices.Finally,The prospects and challenges of 3D printing technology in separation science were discussed.
4.Long-term efficacy of oblique lateral interbody fusion combined with vertebral screw fixation for the treatment of lumbar specific infection
Wen-Hui ZHANG ; Yang-Yang DOU ; Feng-Guang YANG ; Zong-Ru HE ; Yu-Ping YANG ; Li-Zhen FAN ; Qing-Hao CHENG ; Jie LIU ; Hui-Ping TAI
China Journal of Orthopaedics and Traumatology 2024;37(9):893-898
Objective To explore medium and long term efficacy of oblique lateral interbody fusion(OLIF)in treating lumbar specific infection.Methods From October 2017 to January 2021,24 patients with lumbar specific infection were treated by OLIF combined with vertebral screw internal fixation,including 15 males and 9 females,aged from 27 to 61 years old with an average of(43.0±15.0)years old;the courses of disease ranged from 6 to 24 months with an average of(14.0±7.0)months;7 patients with L2-L3,12 patients with L3-L4 and 5 patients with L4-L5;19 patients with tuberculosis infection and 5 patients with brucella infection.The amount of intraoperative blood loss,operative time and complications were recorded,and erythro-cyte sedimentation rate(ESR),C-reactive protein(CRP),visual analogue scale(VAS),Japanese Orthopaedic Association(JOA)score and American Spinal Injury Association(ASIA)rating were compared before and one month after opertaion.Re-sults All patients were followed up from 9 to 24 months with an average of(13.0±6.0)months.Operative time was(132.5±21.4)min,and intraoperative blood loss was(227.3±43.1)ml.ESR and CRP were decreased from(82.34±18.62)mmol·h-1 and(53.08±21.84)mg·L-1 before operation to(33.52±17.31)mmol·h-1 and(15.48±8.36)mg·L-1 at one month after opera-tion,respectively(P<0.05).VAS was decreased from(7.52±1.36)before opertaion to(1.74±0.87)at one month after opera-tion(P<0.05).JOA was increased from(17.86±3.95)before operation to(24.72±3.19)at one month after operation(P<0.05).Four patients had neurological symptoms before operation,and were classified to grade D according to ASIA classifica-tion,who were recovered to grade E at 1 month after operation.One patient was suffered from psoas major muscle injury after operation,and returned to normal at 3 weeks.One patient was suffered from abdominal distension and difficulty in defecation,and relieved after gastrointestinal decompression and enema.No complications such as abdominal organ injury and poor wound healing occurred in all patients.Conclusion OLIF combined with vertebral screw internal fixation is a new minimally invasive surgical method for the treatment of lumbar specific infection,especially the lesion located on the middle lumbar vertebra.It has advantages of less trauma,short operation time,less blood loss,convenient operation,complete removal of the lesion,safety and effectiveness,and has good medium-and long-term efficacy for lumbar specific infection.
5.Novel Immune-related Proteins Identified from Mytilus coruscus by Hemocytes Full-length Transcriptome and Serum Differential Proteome
Wen-Hui XIAO ; Hao-Dong WANG ; Zong-Xin YANG ; Fang SONG ; Yue WANG ; Jian-Yu HE ; Xiao-Lin ZHANG ; Xiao-Jun YAN ; Zhi LIAO
Chinese Journal of Biochemistry and Molecular Biology 2024;40(7):947-963
Mytilus is one of bivalves with great economic and ecological values.The innate immune de-fense of Mytilus shows great significance in the study of marine biological immunology.Hemolymph is the main immune tissue for Mytilus.The Nanopore full-length transcriptome of Mytilus coruscus hemocytes,and the serum differential proteomics based on SDS-PAGE analysis were performed to identify key pro-teins involving in the immune response of Mytilus hemolymph in response of different bacteria and fungi stresses.A total of 44 proteins were identified in the serum induced by different microorganisms.Among them,26 proteins showed significant differential expression level in response to different microbial stres-ses,and their functions were involved in protein folding protection,cell autophagy and apoptosis regula-tion,reactive oxygen species production,energy metabolism regulation,cell detoxification,and immune regulation.The changes in expression levels of these proteins varied in response to different bacterial and fungal stresses,suggesting that Mytilus has different immune response strategies to different bacterial and fungal stresses.The results provide a new scientific basis for understanding the differential immune mech-anism of Mytilus innate immune system in response to different types of microbial invasion,as well as the screening of specific biomarker proteins for microbial infection,and provide ideas for the healthy develop-ment and disease prevention of shellfish aquaculture.
6.History and present situation of seromuscular flap anastomosis in proximal gastrectomy.
Lin Guang FAN ; Ke Chang ZHANG ; Peng CUI ; Liang ZONG ; Wei WEI ; Jie WANG ; Qi Sheng CHENG ; Jin Jie ZHANG ; Yong LIU ; Yin Hao YANG ; Mao Jie ZHANG ; Wen Qing HU
Chinese Journal of Gastrointestinal Surgery 2022;25(5):466-470
With the increasing detection rate of early upper gastric cancer and adenocarcinoma of esophagogastric junction, the safety of proximal gastrectomy with clear indications has been verified, and function-preserving proximal gastrectomy has been widely used. However, proximal gastrectomy destructs the normal anatomical structure of esophagogastric junction, resulting in severe postoperative gastroesophageal reflux symptoms and seriously affecting the quality of life. Among various anti-reflux surgery methods, reconstruction of "cardiac valve" has always been the focus of relevant scholars because its similarity with the mechanism of normal anti-reflux. After years of development, evolution and optimization, the designed seromuscular flap anastomosis includes tunnel muscle flap anastomosis, Hatafuku valvuloplasty, single muscle flap anastomosis and double muscle flap anastomosis. The double muscle flap anastomosis has become a research hotspot because it shows good anti-reflux effect in clinical application. This paper reviews the history, research status and hot issues of seromuscular flap anastomosis of esophageal remnant stomach at home and abroad.
Anastomosis, Surgical/methods*
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Esophagogastric Junction/surgery*
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Gastrectomy/methods*
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Gastroesophageal Reflux/surgery*
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Humans
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Quality of Life
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Stomach Neoplasms/surgery*
7.Transcriptional Modification and Potential Intracellular Signaling Mechanisms in Human Macrophages Primed by Interferon-γ.
Bei LIU ; Hong-Hao GAO ; Li CHENG ; Jia-Le ZHANG ; Yan-Xin DONG ; Shun XIE ; Wen-Rong HUANG ; Shun-Zong YUAN
Journal of Experimental Hematology 2022;30(5):1590-1596
OBJECTIVE:
To explore the transcriptional gene expression profile up-regulated in human macrophages stimulated by interferon-γ (IFN-γ) and the underlying intracellular signaling mechanisms.
METHODS:
RNA-seq was used to sequence and compare the differential gene expression profiles of human macrophage cell line U937 before and after IFN-γ stimulation, and the significantly up-regulated genes were screened out, which were verified by fluorescence-based real-time quantitative polymerase chain reaction (qPCR) in U937 and THP1 cell lines, respectively. JAK/STAT, MAPK/ERK and PI3K/AKT pathway inhibitors were added to simultaneously to the cultured U937 cells upon IFN-γ priming to detect their effects on the expressions of the up-regulated genes to explore the key regulatory mechanisms.
RESULTS:
RNA-seq and qPCR results showed that, the well-recognized chemokines CXCL9, CXCL10 and CXCL11, the APOL family including APOL1, APOL2, APOL3, APOL4, APOL6 and GBP family GBP1, GBP2, GBP3, GBP4 and GBP5 as well were significantly up-regulated in IFN-γ-stimulated U937 cells. JAK/STAT3 pathway inhibitor inhibited the upregulation of APOL1, APOL4, GBP1, GBP4 and GBP5 genes induced by IFN-γ, while MAPK/ERK pathway inhibitor inhibited the upregulation of CXCL10 gene. PI3K/AKT pathway inhibitor inhibited the upregulation of APOL1,APOL4, APOL6, GBP1 and GBP5 genes induced by IFN-γ, all three signal pathway inhibitors could inhibit the upregulation of CXCL9 gene, and none of them could inhibit the upregulation of APOL3 gene.
CONCLUSION
Upon IFN-γ stimulation, some family molecules of APOL and GBP in macrophages are significantly up-regulated, and PI3K/AKT, JAK/STAT3 and MAPK/ERK pathways have positive regulation on their expressions, respectively.
Apolipoprotein L1/pharmacology*
;
Humans
;
Interferon-gamma/pharmacology*
;
Macrophages/metabolism*
;
Phosphatidylinositol 3-Kinases/metabolism*
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Proto-Oncogene Proteins c-akt/metabolism*
;
Signal Transduction
8.3- to 24-month Follow-up on COVID-19 with Pulmonary Tuberculosis Survivors after Discharge: Results from a Prospective, Multicenter Study
Ya Jing WANG ; Yu Xing ZONG ; Hui Gui WU ; Lin Yuan QI ; Zhen Hui LI ; Yu Xin JI ; Lin TONG ; Lei ZHANG ; Bo Ming YANG ; Ye Pu YANG ; Ke Ji LI ; Rong Fu XIAO ; Song Lin ZHANG ; Hong Yun HU ; De Hong LIU ; Fang Shou XU ; Sheng SUN ; Wei WU ; Ya MAO ; Qing Min LI ; Hua Hao HOU ; Yuan Zhao GONG ; Yang GUO ; Wen Li JIAO ; Jin QIN ; Yi Ding WANG ; Fang WANG ; Li GUAN ; Gang LIN ; Yan MA ; Ping Yan WANG ; Nan Nan SHI
Biomedical and Environmental Sciences 2022;35(12):1091-1099
Objective Coronavirus disease 2019 (COVID-19) and tuberculosis (TB) are major public health and social issues worldwide. The long-term follow-up of COVID-19 with pulmonary TB (PTB) survivors after discharge is unclear. This study aimed to comprehensively describe clinical outcomes, including sequela and recurrence at 3, 12, and 24 months after discharge, among COVID-19 with PTB survivors. Methods From January 22, 2020 to May 6, 2022, with a follow-up by August 26, 2022, a prospective, multicenter follow-up study was conducted on COVID-19 with PTB survivors after discharge in 13hospitals from four provinces in China. Clinical outcomes, including sequela, recurrence of COVID-19, and PTB survivors, were collected via telephone and face-to-face interviews at 3, 12, and 24 months after discharge. Results Thirty-two COVID-19 with PTB survivors were included. The median age was 52 (45, 59) years, and 23 (71.9%) were men. Among them, nearly two-thirds (62.5%) of the survivors were moderate, three (9.4%) were severe, and more than half (59.4%) had at least one comorbidity (PTB excluded). The proportion of COVID-19 survivors with at least one sequela symptom decreased from 40.6% at 3 months to 15.8% at 24 months, with anxiety having a higher proportion over a follow-up. Cough and amnesia recovered at the 12-month follow-up, while anxiety, fatigue, and trouble sleeping remained after 24 months. Additionally, one (3.1%) case presented two recurrences of PTB and no re-positive COVID-19 during the follow-up period. Conclusion The proportion of long symptoms in COVID-19 with PTB survivors decreased over time, while nearly one in six still experience persistent symptoms with a higher proportion of anxiety. The recurrence of PTB and the psychological support of COVID-19 with PTB after discharge require more attention.
9.Impact of surgical treatment for locally recurrent rectal cancer on the quality of life of patients.
Min Wei ZHOU ; Zhen Yang LI ; Xiao Dong GU ; Yi Ming ZHOU ; Yi Wen ZANG ; Yi YANG ; Zi Hao WANG ; Zong You CHEN ; Jian Bin XIANG
Chinese Journal of Gastrointestinal Surgery 2021;24(4):352-358
Objective: To investigate the impact of surgical treatment on quality of life in patients with locally recurrent rectal cancer (LRRC). Methods: A descriptive case series study was performed. The complete clinical data of 62 patients who met the diagnostic criteria of LRRC and treated by surgical procedures in Huashan Hospital of Fudan University from January 2012 to November 2019 were analyzed retrospectively. All the patients were followed up at least 12 months. Assessments of urinary function, sexual function, mobility function of lower limb and quality of life were documented. Patients with distant metastasis and surgical history of the urinary system were excluded. According to the criteria of Memorial Sloan Kettering Cancer Center (MSKCC), recurrence were divided into central (n=27), anterior (n=20), posterior (n=7), and lateral (n=8) subtypes. Baseline characteristics, surgical procedures and short-term complications were analyzed. International prostate symptom score (IPSS) and grade of voiding dysfunction were used to evaluate the urinary function. Higher score of IPSS and higher grade of voiding dysfunction indicated worse voiding function. Sexual function for both genders was assessed preoperatively and postoperatively. International index of erectile function-5 (IIEF-5) was used for assessment of male patients and higher score indicated better function. Female sexual function index (FSFI) was used in females and higher score indicated better function. Short-form health survey with 36 items (SF-36), yielding an 8-scale profile of functional health (physical functioning, role-physical, bodily pain, general health, vitality, social functioning, emotional health and mental health) was used to evaluate the quality of life. The higher score indicated the better quality of life. Results: All the operations of 62 patients completed successfully and R0 resection rate was 88.7% (55/62). Postoperative surgical complications occurred in 16 cases (25.8%), including 3 patients of Clavien-Dindo classification III. At postoperative 3-month, 42 patients without ileum cystectomy or ureterostomy suffered from different grade of voiding dysfunction. IPSS increased significantly after the surgery (before surgery: 12.36±4.75, after surgery: 18.40±4.77, t=-9.128, P<0.001). There was no significant difference among the subtypes (P>0.05). At postoperative 12-month, IIEF-5 decreased from 14 (0~25) to 9 (0~19) in males (Z=-5.174, P<0.001) and FSFI deceased from 8.4 (2.0-27.0) to 2.0 (2.0-18.4) in females (Z=-3.522, P<0.001). Scores of physical functioning and role-physical decreased significantly [physical functioning: before surgery 70 (35-85), after surgery 65 (30-80), Z=-3.685, P<0.001; role-physical: before surgery 50 (0-50), after surgery 25(0-75), Z=-4.065, P<0.001], while those of social functioning role-emotional and mental health increased significantly after the surgery [social functioning: before surgery 44 (22-78), after surgery 56 (0-89), Z=-3.509, P<0.001; role-emotional: before surgery 17 (0-100), after surgery 33 (0-100), Z=-2.439, P=0.015; mental health: before surgery 40 (36-76), after surgery 52 (24-80), Z=-3.395, P<0.001]. All surgical procedures decreased the voiding function of LRRC patients and the sexual function of male patients (all P<0.01). However, only total pelvic exenteration and posterior pelvic exenteration decreased FSFI in female patients [before surgery: 8.4 (2.0-27.0) after surgery: 2.0 (2.0-18.4), Z=-2.810, P=0.005]. Conclusions: Multi-visceral resection in LRRC patients may damage voiding and sexual function. However, successful and effective surgical treatment can improve the psychosocial health of LRRC patients.
Female
;
Humans
;
Male
;
Neoplasm Recurrence, Local
;
Quality of Life
;
Rectal Neoplasms/surgery*
;
Rectum
;
Retrospective Studies
10.Application of modified Kamikawa anastomosis in digestive tract reconstruction after proximal gastrectomy.
Liang ZONG ; Peng CUI ; Wei WEI ; Lin Guang FAN ; Jie WANG ; Dong Yang SONG ; Yin Hao YANG ; Mao Jie ZHANG ; Guo Lin HAN ; Wen Qing HU
Chinese Journal of Gastrointestinal Surgery 2021;24(8):691-697
Objective: Traditional Kamikawa anastomosis in digestive tract reconstruction after proximal gastrectomy can greatly decrease the anastomosis-related complications and reduce the incidence of reflux esophagitis, but its complexity limits the wide application. To decrease the complexity of Kamikawa anastomosis, the surgical team of Changzhi People's Hospital of Shanxi Changzhi Medical College improved this technique by using novel notion and reduced surgical procedures. This study aims to evaluate the efficacy and safety of modified Kamikawa anastomosis in digestive tract reconstruction after proximal gastrectomy. Methods: A descriptive cohort study was carried out. Case enrollment criteria: (1) upper gastric carcinoma or esophagogastric junction carcinoma without distant metastasis was confirmed by preoperative gastroscopic biopsy and imaging examination; (2) tumor diameter was less than 4 cm; (3) preoperative clinical staging was cT1-3N1M0. Exclusion criteria: (1) patients received preoperative neoadjuvant chemotherapy; (2) patients had severe heart or lung disease, or poor nutritional status so that they could not tolerate surgery. Clinical data of 25 patients with upper gastric carcinoma or esophagogastric junction carcinoma who underwent modified Kamikawa anastomosis in digestive tract reconstruction in Heji Hospital (8 cases) and Changzhi People's Hospital (17 cases) from April 2019 to December 2020 were retrospectively collected. Of 25 patients, 21 were male and 4 were female, with mean age of 63.0 (49 to 78) years; 3 underwent open surgery and 22 underwent laparoscopic surgery. The modified Kamikawa anastomosis was as follows: (1) the novel notion of total mesangial resection of the esophagogastric junction was applied to facilitate the thorough removal of lymph nodes and facilitate hand-sewn anastomosis and embedding; (2) the diameter of the anastomotic stoma was selected according to the diameter of the esophageal stump, between 2.5 and 3.5 cm, to reduce the occurrence of anastomotic stenosis; (3) an ultrasonic scalpel was used to incise the esophageal stump, which could not only prevent bleeding of the esophageal stump, but also closely seal the esophageal mucosa, muscle layer and serosa to prevent esophageal mucosa retraction; (4) barbed suture was used to suture the remnant stomach fundus and esophagus to fix the stomach fundus in order to reduce the cumbersome and difficult intermittent sutures in a small space; (5) two barbed sutures were used to continuously suture the front and back walls of the anastomosis and complete the suture and fixation of the muscle flap. Relevant indicators of surgical safety, postoperative complications (using the Clavien-Dindo classification), esophageal reflux symptoms and the occurrence of esophagitis (using Los Angeles classification) were analyzed. The gastroesophageal reflux disease (GERD) score, gastroscopy, multi-position digestive tract radiography during postoperative follow-up were used to evaluate the residual gastric motility and anti-reflux efficacy. Results: Modified Kamikawa anastomosis in digestive tract reconstruction after proximal gastrectomy was successfully performed in 25 patients. The surgical time was (5.8±1.8) hours, the intraoperative blood loss was (89.2±11.8) ml, and the average hospital stay was (13.8±2.9) days. Three cases (12.0%) developed postoperative anastomotic stenosis as Clavien-Dindo grade III and were healed after endoscopic dilation treatment. Postoperative upper gastrointestinal radiography showed 1 case (4.0%) with reflux symptoms as Clavien-Dindo grade I. Gastroscopy showed no signs of reflux esophagitis, and its Los Angeles classification was A grade. No anastomotic bleeding, local infection and death were found in all the patients. At postoperative 6-month of follow-up, GERD score showed no significant difference compared to pre-operation (2.7±0.6 vs. 2.4±1.0, t=-1.495, P=0.148). Conclusion: Modified Kamikawa anastomosis in digestive tract reconstruction after proximal gastrectomy is safe and feasible with good anti-reflux efficacy.
Aged
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Anastomosis, Surgical
;
Cohort Studies
;
Esophagogastric Junction/surgery*
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Female
;
Gastrectomy
;
Humans
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Male
;
Middle Aged
;
Retrospective Studies

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