1.EGR2 maintains neuropathic pain by promoting microglial phagocytosis.
Caiyun XI ; Jianxi ZHANG ; Zhifeng HUANG ; Liqiong HE ; Kailu ZOU ; Xiaoping XU ; Qulian GUO ; Bei SUN ; Changsheng HUANG
Journal of Central South University(Medical Sciences) 2025;50(4):586-601
OBJECTIVES:
Neuropathic pain (NP) is one of the most common forms of chronic pain, yet current treatment options are limited in effectiveness. Peripheral nerve injury activates spinal microglia, altering their inflammatory response and phagocytic functions, which contributes to the progression of NP. Most current research on NP focuses on microglial inflammation, with relatively little attention to their phagocytic function. Early growth response factor 2 (EGR2) has been shown to regulate microglial phagocytosis, but its specific role in NP remains unclear. This study aims to investigate how EGR2 modulates microglial phagocytosis and its involvement in NP, with the goal of identifying potential therapeutic targets.
METHODS:
Adult male Sprague-Dawley (SD) rats were used to establish a chronic constriction injury (CCI) model of the sciatic nerve. Pain behaviors were assessed on days 1, 3, 7, 10, and 14 post-surgery to confirm successful model induction. The temporal and spatial expression of EGR2 in the spinal cord was examined using real-time quantitative PCR (RT-qPCR), Western blotting, and immunofluorescence staining. Adeno-associated virus (AAV) was used to overexpress EGR2 in the spinal cord, and behavioral assessments were performed to evaluate the effects of EGR2 modulation of NP. CCI and lipopolysaccharide (LPS) models were established in animals and microglial cell lines, respectively, and changes in phagocytic activity were measured using RT-qPCR and fluorescent latex bead uptake assays. After confirming the involvement of microglial phagocytosis in NP, AAV was used to overexpress EGR2 in both in vivo and in vitro models, and phagocytic activity was further evaluated. Finally, eukaryotic transcriptome sequencing was conducted to screen differentially expressed mRNAs, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses to identify potential downstream effectors of EGR2.
RESULTS:
The CCI model successfully induced NP. Following CCI, EGR2 expression in the spinal cord was upregulated in parallel with NP development. Overexpression of EGR2 via spinal AAV injection enhanced microglial phagocytic activity and increased pain hypersensitivity in rats. Both animal and cellular models showed that CCI or LPS stimulation enhanced microglial phagocytosis, which was further amplified by EGR2 overexpression. Transcriptomic analysis of spinal cord tissues from CCI rats overexpressing EGR2 revealed upregulation of numerous genes associated with microglial phagocytosis and pain regulation. Among them, Lag3 emerged as a potential downstream target of EGR2.
CONCLUSIONS
EGR2 contributes to the maintenance of NP by enhancing microglial phagocytosis in the spinal dorsal horn.
Animals
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Microglia/metabolism*
;
Phagocytosis/physiology*
;
Rats, Sprague-Dawley
;
Neuralgia/physiopathology*
;
Early Growth Response Protein 2/metabolism*
;
Male
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Rats
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Spinal Cord/metabolism*
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Sciatic Nerve/injuries*
2.Thirteen serum biochemical indexes and five whole blood coagulation indices in a point-of-care testing analyzer: ideal protocol for evaluating pulmonary and critical care medicine.
Mingtao LIU ; Li LIU ; Jiaxi CHEN ; Zhifeng HUANG ; Huiqing ZHU ; Shengxuan LIN ; Weitian QI ; Zhangkai J CHENG ; Ning LI ; Baoqing SUN
Journal of Zhejiang University. Science. B 2025;26(2):158-171
The accurate and timely detection of biochemical coagulation indicators is pivotal in pulmonary and critical care medicine. Despite their reliability, traditional laboratories often lag in terms of rapid diagnosis. Point-of-care testing (POCT) has emerged as a promising alternative, which is awaiting rigorous validation. We assessed 226 samples from patients at the First Affiliated Hospital of Guangzhou Medical University using a Beckman Coulter AU5821 and a PUSHKANG POCT Biochemistry Analyzer MS100. Furthermore, 350 samples were evaluated with a Stago coagulation analyzer STAR MAX and a PUSHKANG POCT Coagulation Analyzer MC100. Metrics included thirteen biochemical indexes, such as albumin, and five coagulation indices, such as prothrombin time. Comparisons were drawn against the PUSHKANG POCT analyzer. Bland-Altman plots (MS100: 0.8206‒0.9995; MC100: 0.8318‒0.9911) evinced significant consistency between methodologies. Spearman correlation pinpointed a potent linear association between conventional devices and the PUSHKANG POCT analyzer, further underscored by a robust correlation coefficient (MS100: 0.713‒0.949; MC100: 0.593‒0.950). The PUSHKANG POCT was validated as a dependable tool for serum and whole blood biochemical and coagulation diagnostics. This emphasizes its prospective clinical efficacy, offering clinicians a swift diagnostic tool and heralding a new era of enhanced patient care outcomes.
Humans
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Point-of-Care Testing
;
Critical Care
;
Blood Coagulation Tests/methods*
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Male
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Blood Coagulation
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Female
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Middle Aged
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Reproducibility of Results
;
Prothrombin Time
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Aged
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Adult
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Point-of-Care Systems
3.Meta-analysis of sealants versus fluoride varnishes for the prevention of occlusal surface caries in children's first permanent molars.
Jingya HAN ; Yajun ZHANG ; Mengzhen JI ; Jingfei SUN ; Shuhan JIA ; Zhifeng WANG
West China Journal of Stomatology 2025;43(3):383-394
OBJECTIVES:
To assess the effectiveness of the comparison between pit and fissure sealants and fluoride varnishes, as well as various types of sealants, in preventing caries on the occlusal surface of children's first permanent molars (FPM).
METHODS:
Conduct a comprehensive search of literature published between January 1, 1988, and May 30, 2024, in the following databases: China National Knowledge Infrastructure, Web of Science, Cochrane Library, Embase, PubMed, China Science Periodical Database and China Biology Medicine database. Meta-analysis and subgroup analyses were performed on the literature that met the inclusion criteria.
RESULTS:
A total of 5 618 pieces of literature were retrieved, resulting in the inclusion of 14 in the study. Meta-analysis showed that there was no statistically significant difference in the efficacy between varies pit and fissure sealants compared to fluoride varnishes, and between varies types of sealants in preventing caries on the occlusal surface of children's first permanent molars within 24 months post-surgery (P>0.05).
CONCLUSIONS
Within 24 months, there was no significant difference in the effectiveness of using resin-based or glass iomomer pit and fissure sealants compared with fluoride varnishes in preventing occlusal caries in FPM in children; within 24 months, there was no significant difference in the effectiveness of using resin-based sealants compared with ART sealants in preventing occlusal caries in FPM in children. ART sealants are recommended over resin-based sealers for children who have no conditions for chair-side manipulation or who are poorly co-operative.
Humans
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Pit and Fissure Sealants/therapeutic use*
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Dental Caries/prevention & control*
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Molar
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Child
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Fluorides, Topical/therapeutic use*
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Dentition, Permanent
4.Mechanistic insights into “Three Methods and Three Acupoints” Tuina therapy for improving spinal microcirculation and motor function in sciatic nerve injury model rats
Hanyu Zhang ; Yingqi Zhang ; Hourong Wang ; Jiayue Liu ; Jiawei Sun ; Jinping Chen ; Zhifeng Liu ; Tianyuan Yu ; Jian Shu
Journal of Traditional Chinese Medical Sciences 2025;2025(1):125-134
Objective:
To investigate the effects of “Three Methods and Three Acupoints” (TMTP) Tuina therapy on spinal microcirculation in sciatic nerve injury (SNI).
Methods:
Thirty-six Sprague–Dawley rats were randomly assigned to four groups: normal, sham operation, model, and TMTP Tuina. Successful model induction was confirmed by observable hind limb lameness. After 20 sessions, hind limb grip strength and motor nerve conduction velocity (MNCV) were measured at baseline and following the 10th and 20th intervention. CD31 and α-SMA in the ventral horn of SNI model rats were detected using immunofluorescence. Motor neurons in the ventral horn were detected by Nissl staining. PTEN levels in the ventral horn were measured by ELISA, and PI3K, Akt, BDNF, VEGF, and HIF-1α expression was determined by RT-PCR. Spinal cord microcirculation was evaluated by western blotting analysis of the levels of Akt, p-Akt, BDNF, and VEGF.
Results:
Hind limb grip strength and MNCV significantly improved in the TMTP Tuina group compared to the model group (both P < .001). Morphology of ventral horn motor neurons in the TMTP Tuina group improved compared to the model group, with increased expressions of α-SMA (P = .002) and CD31 (P = .006). Western blot analysis indicated increased expression of VEGF (P = .005), p-Akt (P < .001), and BDNF (P = .008) in the ventral horn following Tuina treatment. RT-PCR analysis revealed increased expression of PI3K, Akt, BDNF, VEGF and HIF-1α (all P < .05). In contrast, expression of PTEN decreased compared to the model group (P < .001).
Conclusion
TMTP Tuina therapy may restore motor function in rats, enhance ventral horn motor neuron morphology, and promote angiogenesis and vascular smooth muscle proliferation. The mechanism may involve the activation of the PI3K/Akt signaling pathway.
5.Mechanism of the immediate analgesic effect of the"three methods and three points"tuina technique based on the IL-17F/IL-17RC signaling pathway and M1 microglia
Jinping CHEN ; Zhifeng LIU ; Tianyuan YU ; Hourong WANG ; Yingqi ZHANG ; Qian GUAN ; Yajing XU ; Zhenjie YANG ; Chula SA ; Runlong ZHANG ; Hanyu ZHANG ; Jiayue LIU ; Jiawei SUN
Journal of Beijing University of Traditional Chinese Medicine 2024;47(1):116-123
Objective By observing the effects of"three methods and three points"tuina technique on the expression of interleukin-17F(IL-17F),interleukin-17 receptor C(IL-17RC),activator 1 of nuclear transcription factor-κB(Act1),tumour necrosis factor receptor-associated factor 6(TRAF6)and M1 microglial cell expression in the spinal dorsal horn of rats with mild chronic compressive injury(minor CCI)model,we explored the immediate analgesic mechanism of tuina on peripheral neuropathic pain(pNP).Methods Thirty-six SD rats were divided into the sham group,the model group and the tuina group according to the random number method,twelve rats in each group,and the minor CCI model was replicated by ligating the right sciatic nerve.The rats in the tuina group were subjected to pointing,plucking and kneading at the BL37,BL57 and GB34 points on the affected side using a tuina simulator,while the sham group and the model group were only grasped and restrained,and were intervened for one time.The mechanical pain test and cold plate test were used to evaluate the response of rats to mechanical stimulation and cold stimulation after immediate intervention.The protein expression of IL-17F and TRAF6 in the spinal dorsal horn of rats in each group was detected by Western blotting.The mRNA expression of IL-17F,IL-17RC,Act1 and TRAF6 in the spinal dorsal horn of rats in each group was detected by real-time PCR.The average fluorescence intensity of M1 microglia in the spinal dorsal horn of rats in each group was detected by immunofluorescence.Results Behavioral results showed that before intervention,compared with the sham group,paw mechanical withdraw threshold(PMWT)decreased and cold sensitivity threshold(CST)increased in the model group and the tuina group;after tuina intervention,PMWT in the tuina group was increased,and CST was decreased compared with the model group;after intervention,PMWT in the tuina group was increased,while CST was decreased(P<0.05).RT-PCR results showed that compared with the sham group,mRNA expression levels of IL-17F,IL-17RC,TRAF6 and Act1 in the spinal dorsal horn of the model group were increased;compared with model group,the mRNA expression levels of above indexes in the tuina group were decreased(P<0.05).Western boltting results showed that compared with the sham group,the expression levels of IL-17F and TRAF6 protein in the spinal dorsal horn of the model group were increased;compared with the model group,the expression levels of IL-17F and TRAF6 protein in the tuina group decreased(P<O.05).Immunofluorescence results showed that the mean fluorescence intensity of CD40 in the spinal dorsal horn of model group was enhanced compared with the sham group;compared with the model group,the mean fluorescence intensity of CD40 in the tuina group was decreased(P<0.05).Conclusion The"three methods and three points"tuina technique can produce immediate analgesia by inhibiting the expression of IL-17F,IL-17RC,Act1,TRAF6 and the activation of M1 microglia in the dorsal horn of the spinal cord after one intervention.
6.Furry animal allergen components diagnosis: identification of main components and clinical management strategies
Zhifeng HUANG ; Aoli LI ; Huiqing ZHU ; Ziyu YIN ; Baoqing SUN
Chinese Journal of Preventive Medicine 2024;58(6):931-940
Furry animal allergens, particularly cat and dog hair and dander, are common allergens in indoor environments, affecting the health of people world widely. Key sensitizing components such as Fel d 1 from cats and Can f 1 from dogs have been extensively studied and identified by the scientific community. Component resolved diagnosis (CRD) technology in modern diagnostic methods provides an accurate way to identify and distinguish these components, which is extremely important for the prevention of furry animal allergies and the formulation of personalized treatment strategies. To enhance the understanding of furry animal component diagnosis and promote the alignment of the Chinese discipline of allergology with international standards, this article interprets and explains the content of the "Molecular Allergology User′s Guide 2.0" recently released by the European Academy of Allergy and Clinical Immunology. It focuses on the epidemiological characteristics of furry animal components, the diversity of allergen protein families, and their clinical diagnosis and management.
7.Design and application of clinical probation log based on subjective-objective-assessment-plan
Zhifeng WU ; Li MING ; Zhangya YANG ; Min WANG ; Yizhen SUN ; Wang YANG ; Zhiqiang CHEN ; Zhangxue HU ; Yuping ZHANG
Chinese Journal of Medical Education Research 2024;23(7):906-909
Objective:To investigate the application value of a new clinical probation log designed based on the subjective-objective-assessment-plan (SOAP) structured medical record.Methods:Quantitative and qualitative studies were conducted among 97 students in the autumn semester to evaluate the effect of the new clinical probation log based on the SOAP structured medical record on their academic performance and clinical thinking. SPSS 26.0 was used to perform the t-test and ANOVA. Results:The students using the new version had a significantly better score (96.29±1.38) than those using the old version (93.53±1.60) ( P<0.001), while the multivariate analysis showed that the students who first used the old version and then switched to the new version had a significantly better improvement in the score compared with those who first used the new version and then switched to the old version ( P<0.001). The qualitative interview showed that it was necessary to use handwritten internship log, and compared with the old version, the new version could better promote the ordered clinical thinking of students. The teacher comments could give feedbacks, and all the students interviewed thought that the new version held promise for clinical application. Conclusions:The clinical probation log based on the SOAP structured medical record can help to improve the effectiveness of probation and cultivate clinical thinking ability, and thus it holds promise for application in clinical probation teaching.
8.Clinical characteristics of 11 patients with Vibrio vulnificus infection and the establishment of a rapid diagnosis procedure for this disease
Weipeng LIN ; Xu MU ; Shenghua CHEN ; Chunjing HE ; Hanhua LI ; Chuanwei SUN ; Huining BIAN ; Wen LAI ; Zhifeng HUANG
Chinese Journal of Burns 2024;40(3):266-272
Objective:To analyze the clinical characteristics of patients with Vibrio vulnificus infection, share diagnosis and treatment experience, and establish a rapid diagnosis procedure for this disease. Methods:This study was a retrospective case series study. From January 2009 to November 2022, 11 patients with Vibrio vulnificus infection who met the inclusion criteria were admitted to the Department of Burns and Wound Repair of Guangdong Provincial People's Hospital Affiliated to Southern Medical University. The gender, age, time of onset of illness, time of admission, time of diagnosis, route of infection, underlying diseases, affected limbs, clinical manifestations and signs on admission, white blood cell count, hemoglobin, platelet count, C-reactive protein (CRP), alanine transaminase (ALT), aspartate transaminase (AST), creatinine, procalcitonin, albumin, N-terminal pro-B-type natriuretic peptide (NT-proBNP), and blood sodium levels on admission, culture results and metagenomic next-generation sequencing (mNGS) results of pathogenic bacteria and the Vibrio vulnificus drug susceptibility test results during hospitalization, treatment methods, length of hospital stay, and outcomes of all patients were recorded. Comparative analysis was conducted on the admission time and diagnosis time of patients with and without a history of exposure to seawater/marine products, as well as the fatality ratio and amputation of limbs/digits ratio of patients with and without early adequate antibiotic treatment. For the survived patients with hand involvement, the hand function was assessed using Brunnstrom staging at the last follow-up. Based on patients' clinical characteristics and treatment conditions, a rapid diagnosis procedure for Vibrio vulnificus infection was established. Results:There were 7 males and 4 females among the patients, aged (56±17) years. Most of the patients developed symptoms in summer and autumn. The admission time was 3.00 (1.00, 4.00) d after the onset of illness, and the diagnosis time was 4.00 (2.00, 8.00) d after the onset of illness. There were 7 and 4 patients with and without a history of contact with seawater/marine products, respectively, and the admission time of these two types of patients was similar ( P>0.05). The diagnosis time of patients with a history of contact with seawater/marine products was 2.00 (2.00, 5.00) d after the onset of illness, which was significantly shorter than 9.00 (4.25, 13.00) d after the onset of illness for patients without a history of contact with seawater/marine products ( Z=-2.01, P<0.05). Totally 10 patients had underlying diseases. The affected limbs were right-hand in 8 cases, left-hand in 1 case, and lower limb in 2 cases. On admission, a total of 9 patients had fever; 11 patients had pain at the infected site, and redness and swelling of the affected limb, and 9 patients each had ecchymosis/necrosis and blisters/blood blisters; 6 patients suffered from shock, and 2 patients developed multiple organ dysfunction syndrome. On admission, there were 8 patients with abnormal white blood cell count, hemoglobin, and albumin levels, 10 patients with abnormal CRP, procalcitonin, and NT-proBNP levels, 5 patients with abnormal creatinine and blood sodium levels, and fewer patients with abnormal platelet count, ALT, and AST levels. During hospitalization, 4 of the 11 wound tissue/exudation samples had positive pathogenic bacterial culture results, and the result reporting time was 5.00 (5.00, 5.00) d; 4 of the 9 blood specimens had positive pathogenic bacterial culture results, and the result reporting time was 3.50 (1.25, 5.00) d; the mNGS results of 7 wound tissue/exudation or blood samples were all positive, and the result reporting time was 1.00 (1.00, 2.00) d. The three strains of Vibrio vulnificus detected were sensitive to 10 commonly used clinical antibiotics, including ciprofloxacin, levofloxacin, and amikacin, etc. A total of 10 patients received surgical treatment, 4 of whom had amputation of limbs/digits; all patients received anti-infection treatment. The length of hospital stay of 11 patients was (26±11) d, of whom 9 patients were cured and 2 patients died. Compared with that of the 6 patients who did not receive early adequate antibiotic treatment, the 5 patients who received early adequate antibiotic treatment had no significant changes in the fatality ratio or amputation of limbs/digits ratio ( P>0.05). In 3 months to 2 years after surgery, the hand function of 8 patients was assessed, with results showing 4 cases of disabled hands, 2 cases of incompletely disabled hands, and 2 cases of recovered hands. When a patient had clinical symptoms of limb redness and swelling and a history of contact with seawater/marine products or a pre-examination triage RiCH score of Vibrio vulnificus sepsis ≥1, the etiological testing should be initiated immediately to quickly diagnose Vibrio vulnificus infection. Conclusions:Vibrio vulnificus infection occurs most frequently in summer and autumn, with clinical manifestations and laboratory test results showing obvious infection characteristics, and may be accompanied by damage to multiple organ functions. Both the fatality and disability ratios are high and have a great impact on the function of the affected limbs. Early diagnosis is difficult and treatment is easily delayed, but mNGS could facilitate rapid detection. For patients with red and swollen limbs accompanied by a history of contact with seawater/marine products or with a pre-examination triage RiCH score of Vibrio vulnificus sepsis ≥1, the etiological testing should be initiated immediately to quickly diagnose Vibrio vulnificus infection.
9.A study on quantification assessment of segmental volume after thigh liposuction with three-dimensional scanning technology
Yifei LI ; Zhifeng LI ; Yunpeng GU ; Yicheng LIU ; Qianwen LYU ; Jingjing SUN ; Yue QI ; Guie MA
Chinese Journal of Plastic Surgery 2024;40(6):597-604
Objective:To investigate the quantification assessment of segmental volume after thigh liposuction utilizing three-dimensional(3D) scanning technology.Methods:This retrospective study was performed with the analysis of 3D scanning images of patients who had undergone bilateral thigh liposuction in Body Contouring & Fat Grafting Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2018 to September 2022. Preoperative and postoperative 3D scanning were performed to build visual 3D models of bilateral thighs. From top to the bottom, bilateral thighs were segmented into seven sections with a space of 5 cm in acquired 3D model. Certain measurements and calculation of preoperative and postoperative volume and volume change rate of the overall thigh and each segment were conducted, to validate the efficiency of liposuction (EOL). Additionally, EOL of each thigh segment was computed and the symmetry of bilateral thighs was analyzed before and after surgery. The volume differences were computed using the paired Wilcoxon rank-sum test, and intraclass correlation coefficient (ICC) was exerted to assess the symmetry of bilateral thighs before and after surgery.Results:A total of 36 female patients were included in the study, with an age range of 18 to 49 years and a mean age of (28.6±7.6) years. Follow-up duration ranged from 23 to 1 133 days postoperatively, with a mean follow-up period of 274.7 days. The results of 3D scanning measurements indicated significant changes ( P<0.01) in both the overall volume of the thigh and the volumes of each segment before and after surgery. The highest volume change rate and EOL were observed in the uppermost segment, and the volumetric change rate and EOL exhibited a descending trend across the segments of the thigh from the uppermost to the lowermost segments. The ICC of the volume of each segment consistently surpassed 0.950 whether preoperatively or postoperatively, indicating a high level of symmetry between the bilateral thighs, and the ICC of overall volume showed a notable increase from 0.992 preoperatively to 0.997 postoperatively. Conclusion:3D scanning technology can be exerted to quantify the volume changes before and after thigh liposuction. This study provided quantitative and objective evidence to confirm the efficacy of thigh liposuction procedure, elucidating that the most significant liposuction effects observed in the uppermost segment of the thigh. Moreover, postoperative assessments reveal a further enhancement in bilateral thigh symmetry.
10.Comparison of the anorectal function before and after neoadjuvant radiotherapy in mid-low rectal cancer: a retrospective observational study from single center
Zhen SUN ; Zhifeng WANG ; Xiyu SUN ; Lai XU ; Guannan ZHANG ; Junyang LU ; Yi XIAO
Chinese Journal of Gastrointestinal Surgery 2024;27(1):63-68
Objective:The aim of this study was to evaluate the impact of neoadjuvant radiotherapy on anorectal function of patients with mid-low rectal cancer by means of high-resolution anorectal manometry.Methods:A retrospective observational study was conducted. Information on patients with mid-low rectal cancer was collected from the prospective registry database of Rectal Cancer at Peking Union Medical College Hospital (PUMCH) from June 2020 to April 2023. Anorectal functions were detected using three-dimensional high-resolution manometry system. Logistic regression analysis was performed to identify the factors associated with the changed anorectal manometry.Results:A total of 45 patients with mid-low rectal cancer were included in the study. Thirty-two (71.1%) patients were male, 13 (28.9%) patients were female. The mean age was 60±11 years, and the mean BMI was 23.4±3.7 kg/m 2. The mean distance between the lower edge of the tumor and the anal verge was 5.4±1.5 cm. The median size of the tumor was 3.4 (2.9-4.5) cm, and the median circumferential extent of the tumor was 66.0 (45.5-75.0) %. 41 (81.1%) patients were MRI T3-4 and 40 (88.9%) patients were MRI N positive. The resting pressure has a decreasing trend after neoadjuvant radiotherapy (55.3±32.0 mmHg vs. 48.0±28.5 mmHg, t=1.930, P=0.060). There was no significant change in maximum squeezing and the length of the high-pressure zone after neoadjuvant radiotherapy. All volumes describing rectal sensitivity (first sensation, desire to defecate, and maximum tolerance) were lower after neoadjuvant radiotherapy. And maximum tolerance was significantly lower (66.0 [49.0,88.0] ml vs. 52.0 [39.0,73.5] ml, Z=-2.481, P=0.013). Univariate analysis demonstrated that the downstage of N-stage was associated with the decrease in maximum tolerance (OR=6.533, 95%CI:1.254-34.051, P=0.026). Conclusion:Neoadjuvant radiotherapy damages anorectal function by decreasing the resting pressure and rectal sensory threshold of patients. The N-stage downstaging was associated with a decrease in maximum tolerance.


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