Objective: To explore the changes of mechanosensitive channels Piezos (Piezo 1 and Piezo 2) in neurogenic bladder (NB) of young rats and their effects，so as to provide reference for clinical search of new therapeutic targets. Methods: A total of 30 female young SD rats were divided into 5 groups based on random number table method：sham operation group (sham)，2-week nerve transection group (NB-2W)，6-week nerve transection group (NB-6W)，2-week nerve transection + Piezos inhibitor group (NB-P-2W) and 6-week nerve transection + Piezos inhibitor group (NB-P-6W)，with 6 rats in each group.The NB models were constructed by transecting the L6 and S1 spinal nerves of young rats.The NB-2W and NB-6W groups were not intervened after modeling，while the NB-P-2W and NB-P-6W groups were intraperitoneally injected with Piezos inhibitor GsMTx4 (10 mg/kg) every 2 days after modeling.Bladder cystometry and ultrasound were performed after 2 and 6 weeks of transection.The expressions of Piezos and fibrosis-related indexes (Collagen Ⅰ and α-smooth muscle actin) were detected in bladder tissues. Results: The results of bladder cystometry showed that the basal bladder pressure in NB-2W group was significantly increased，while it was slightly decreased but was still higher in NB-6W group than in the sham group (P<0.05).Basal bladder pressure was lower in NB-P-2W group than in NB-2W group，but was higher than that in the sham group; basal bladder pressure was lower in NB-P-6W group than in NB-6W group，but higher than that in the sham group (P<0.05).Compared with the sham group，the NB-2W and NB-6W groups had firstly increased and then decreased maximum cystometric capacity (MCC) (P<0.05).Compared with NB-2W group，NB-P-2W group had lower bladder leakage point pressure (BLPP)，but higher MCC and bladder compliance (BC) (P<0.05).Compared with NB-6W group，NB-P-6W group had significantly lower BLPP but higher MCC and BC (P<0.05).HE and MASSON staining and ultrasound results showed that，with the extension of nerve transection time，bladder fibrosis gradually worsened，the bladder wall became rough and thickened，calculi were visible inside，and hydronephrosis gradually appeared; the degree of fibrosis in NB-P-2W and NB-P-6W groups was less than that in NB-2W and NB-6W groups，and no hydronephrosis was observed in the upper urinary tract.In addition，Western blotting and immunohistochemical results showed that NB-2W and NB-6W groups had significantly higher relative expression levels of Piezos，Collagen Ⅰ and α-SMA than the sham group (P<0.01)，while NB-P-2W and NB-P-6W groups had lower relative expression levels of Piezos,Collagen Ⅰ and α-SMA than NB-2W and NB-6W groups (P<0.01). Conclusion: The increased expressions of mechanosensitive channels Piezos in NB young rats may be involved in the progression of bladder fibrosis，but its mechanism needs further study.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a prevalent type of cancer with a high mortality rate in its late stages. One of the major challenges in OSCC treatment is the resistance to epidermal growth factor receptor (EGFR) inhibitors. Therefore, it is imperative to elucidate the mechanism underlying drug resistance and develop appropriate precision therapy strategies to enhance clinical efficacy. METHODS: To evaluate the efficacy of the combination of the Ca 2+ /calmodulin-dependent protein kinase II (CAMK2) inhibitor KN93 and EGFR inhibitors, we performed in vitro and in vivo experiments using two FAT atypical cadherin 1 ( FAT1 )-deficient (SCC9 and SCC25) and two FAT1 wild-type (SCC47 and HN12) OSCC cell lines. We assessed the effects of EGFR inhibitors (afatinib or cetuximab), KN93, or their combination on the malignant phenotype of OSCC in vivo and in vitro . The alterations in protein expression levels of members of the EGFR signaling pathway and SRY-box transcription factor 2 (SOX2) were analyzed. Changes in the yes-associated protein 1 (YAP1) protein were characterized. Moreover, we analyzed mitochondrial dysfunction. Besides, the effects of combination therapy on mitochondrial dynamics were also evaluated. RESULTS: OSCC with FAT1 mutations exhibited resistance to EGFR inhibitors treatment. The combination of KN93 and EGFR inhibitors significantly inhibited the proliferation, survival, and migration of FAT1 -mutated OSCC cells and suppressed tumor growth in vivo . Mechanistically, combination therapy enhanced the therapeutic sensitivity of FAT1 -mutated OSCC cells to EGFR inhibitors by modulating the EGFR pathway and downregulated tumor stemness-related proteins. Furthermore, combination therapy induced reactive oxygen species (ROS)-mediated mitochondrial dysfunction and disrupted mitochondrial dynamics, ultimately resulting in tumor suppression. CONCLUSION Combination therapy with EGFR inhibitors and KN93 could be a novel precision therapeutic strategy and a potential clinical solution for EGFR-resistant OSCC patients with FAT1 mutations.
Humans ; ErbB Receptors/metabolism* ; Mouth Neoplasms/metabolism* ; Cell Line, Tumor ; Animals ; Drug Resistance, Neoplasm/genetics* ; Cadherins/metabolism* ; Carcinoma, Squamous Cell/metabolism* ; Mice ; Mutation/genetics* ; Mice, Nude ; Protein Kinase Inhibitors/therapeutic use* ; Cetuximab/pharmacology* ; Afatinib/therapeutic use* ; Cell Proliferation/drug effects* ; Signal Transduction/drug effects*

The medicinal use of Malvae Semen has a long history. In this paper， by consulting the ancient materia medica， prescription， agronomy， literature and other aspects of the classics， the name， origin， evolution of scientific name， quality， harvesting and processing， functions and indications and others of Malvae Semen were systematically sorted out and verified， so as to provide a basis for the development and utilization of famous classical formulas containing this herb. According to the textual research， Shennong Bencaojing began to use Dongkuizi as the correct name， which was used in the past dynasties， and there were also aliases such as Kuicaizi， Huacai， and Kuizi. Through the original research， it can be seen that Kuicai is the mainstream original plant of Malvae Semen， that is， Malva verticillata var. crispa， the Alcea rosea and M. cathayensis are also used. In modern times， the seeds of Abutilon theophrasti have been passed off as Malvae Semen， while the seeds of M. verticillata var. crispa have rarely been used in medicine. And Abutili Semen has been another medicinal material with different efficacy since the collection of Newly Revised Materia Medica in the Tang dynasty. Since the Ming and Qing dynasties， the cultivation of Kuicai has been decreasing， while A. theophrasti is more common and easy to obtain， and Abutili Semen and Malvae Semen are similar in morphology and confused， which should be corrected. In addition， Malvae Fructus is a Mongolian customary medicinal herb， which is different from the traditional use of seeds in traditional Chinese medicine. Kuicai， as an important vegetable in history， was widely cultivated and gradually shrunk after the Song dynasty， it is now mainly produced in southern provinces. The quality evaluation of Malvae Semen is better for those with dry bodies， full grain， grayish brown color， no mud， and no impurities. The harvesting is generally in the autumn and winter. After drying， it is seeded， sieved peel and impurities， mashed， or slightly stir-fried to yellow-white color with gentle fire. It is sweet， cold and slippery in nature and taste， with the main effects of laxation， diuresis， lactation and elimination of swelling. The efficacy of Abutili Semen is clearing heat and removing toxicity， promoting diuresis and removing nebula， the efficacy is quite different from that of Malvae Semen. Based on the results of textual research， it is suggested that M. verticillata var. crispa should be used as the medicinal source of Malvae Semen in the development of famous classical formulas， the corresponding processing methods should be selected according to the requirements of drug processing in the formulas， while the raw products are recommended to be used if the processing is not specified.

The medicinal use of Malvae Semen has a long history. In this paper， by consulting the ancient materia medica， prescription， agronomy， literature and other aspects of the classics， the name， origin， evolution of scientific name， quality， harvesting and processing， functions and indications and others of Malvae Semen were systematically sorted out and verified， so as to provide a basis for the development and utilization of famous classical formulas containing this herb. According to the textual research， Shennong Bencaojing began to use Dongkuizi as the correct name， which was used in the past dynasties， and there were also aliases such as Kuicaizi， Huacai， and Kuizi. Through the original research， it can be seen that Kuicai is the mainstream original plant of Malvae Semen， that is， Malva verticillata var. crispa， the Alcea rosea and M. cathayensis are also used. In modern times， the seeds of Abutilon theophrasti have been passed off as Malvae Semen， while the seeds of M. verticillata var. crispa have rarely been used in medicine. And Abutili Semen has been another medicinal material with different efficacy since the collection of Newly Revised Materia Medica in the Tang dynasty. Since the Ming and Qing dynasties， the cultivation of Kuicai has been decreasing， while A. theophrasti is more common and easy to obtain， and Abutili Semen and Malvae Semen are similar in morphology and confused， which should be corrected. In addition， Malvae Fructus is a Mongolian customary medicinal herb， which is different from the traditional use of seeds in traditional Chinese medicine. Kuicai， as an important vegetable in history， was widely cultivated and gradually shrunk after the Song dynasty， it is now mainly produced in southern provinces. The quality evaluation of Malvae Semen is better for those with dry bodies， full grain， grayish brown color， no mud， and no impurities. The harvesting is generally in the autumn and winter. After drying， it is seeded， sieved peel and impurities， mashed， or slightly stir-fried to yellow-white color with gentle fire. It is sweet， cold and slippery in nature and taste， with the main effects of laxation， diuresis， lactation and elimination of swelling. The efficacy of Abutili Semen is clearing heat and removing toxicity， promoting diuresis and removing nebula， the efficacy is quite different from that of Malvae Semen. Based on the results of textual research， it is suggested that M. verticillata var. crispa should be used as the medicinal source of Malvae Semen in the development of famous classical formulas， the corresponding processing methods should be selected according to the requirements of drug processing in the formulas， while the raw products are recommended to be used if the processing is not specified.

BACKGROUND:The relationship between rheumatoid arthritis and coronary atherosclerosis has received extensive attention.Inflammation is related to rheumatoid arthritis and coronary atherosclerosis,indicating that there may be a common pathophysiological pathway between the two diseases.However,observational studies have not yet clarified the causal relationship.OBJECTIVE:To explore whether there is a causal relationship between rheumatoid arthritis and coronary atherosclerosis,as well as the potential causal relationship with 1 400 serum metabolites and 91 inflammatory factors through a Mendelian randomization analysis.METHODS:Coronary atherosclerosis data are from Finngen database,rheumatoid arthritis data are from IEU OpenGWAS database,serum metabolites data are from Canadian Longitudinal Study on Aging,Augsburg Cooperative Health Research and British Twin Project Research,and data of 91 inflammatory proteins are from research published in Nature Immunology in 2023.Mendelian randomization analysis was performed using data from genome-wide association studies,and causal effects were evaluated using inverse variance weighting,MR-Egger regression,weighted median,weighted model,and simple model methods,with inverse variance weighting being the primary analysis method.To enhance robustness,Cochran's Q-test MR-Egger intercept was used for sensitivity analysis.RESULTS AND CONCLUSION:(1)Inverse variance weighting results showed that rheumatoid arthritis was positively correlated with the increased relative risk of coronary atherosclerosis(odds ratio=1.002,95％confidence interval=1.001-1.003,P=0.003).There was no reverse causal relationship between coronary atherosclerosis and rheumatoid arthritis.In addition,96 serum metabolites and 9 inflammatory factors were found to have causal relationships with coronary atherosclerosis.There was a causal relationship between 51 serum metabolites and 7 inflammatory factors and rheumatoid arthritis.(2)This study provided epidemiological evidence between rheumatoid arthritis and coronary atherosclerosis,and emphasized the potential role of serum metabolites and inflammatory factors in the pathogenesis of these diseases.These findings may contribute to the development of new treatment strategies.Due to the limited inclusion of data from Asian populations,most contemporary studies used international databases and European population analyses.By collecting and analyzing the health data of European populations,it is conducive to a better understanding of the effects and potential role of Chinese medicine in Europe,and to further promote the practice of modern integration of Western and Chinese medicine.Meanwhile,through the comparative study with the European databases,it is possible to reveal the genetic differences and susceptibility to diseases among different populations,providing more dimensions and perspectives for global health research.

Objective To construct a prediction model for targeted biopsy(TB)of the prostate based on multiparameter magnetic resonance imaging(mp-MRI)and prostate-specific antigen density(PSAD)to predict the outcomes TB in patients with a score of 4-5 on the Prostate Imaging Reporting and Data System(PI-RADS).Methods Clinical data of 669 patients with PI-RADS 4-5 receiving transperineal TB in our hospital during Jan.2022 and Dec.2023 were retrospectively analyzed.The data were divided into the training set and validation set with a ratio of 2∶1.Independent predictors of TB results were identified with univariate and multivariate logistic regression to construct a formula for the prediction model.A prediction model was subsequently constructed and validated using the validation set to assess its efficacy and predictive performance with the area under the receiver operating characteristic curve(AUC).The relative importance of each independent predictor in the formula was analyzed.Results Univariate and multivariate logistic regression analyses showed that age,total number of lesions,histological location,PI-RADS score and PSAD were significantly associated with the TB outcomes(P＜0.05)and could be used as independent predictors,with PI-RADS score and PSAD making the highest contribution to outcome prediction,accounting for 27.59％and 37.58％,respectively.The training set had an AUC of 0.840(95％CI:0.800-0.881),which was more predictive than other single predictors,and the high-risk group based on the optimal threshold of 0.833 increased the positive biopsy rate from 79.3％to 94.4％.The validation set had an AUC of 0.865(95％CI:0.810-0.920),and the high-risk group based on the optimal threshold of 0.594 increased the positive biopsy rate from 80.0％to 96.2％.Conclusion The prediction model has good predictive ability for lesions with PI-RADS 4-5,which can significantly improve the positive detection rate and reduce a large number of unnecessary systematic puncture.

BACKGROUND:The analgesic effect of stagnant moving needle on myofascial pain syndrome is remarkable,but the analgesic mechanism is still unclear.OBJECTIVE:To investigate the analgesic mechanism of stagnant moving needle acupuncture in the treatment of myofascial pain syndrome.METHODS:Fifty-four SD rats were randomly divided into a blank group(n=16)and a modeling group(n=38).The models of leftmyofascial pain syndrome in the modeling group were prepared by using the method of"striking combined with centrifugal movement".Twelve weeks after modeling,six mice were randomly selected to verify the success of the modeling.The rest of the 32 rats were randomly divided into the model group and the stagnant moving needle group,with 16 rats in each group.The stagnant needle moving group was treated by stagnant moving needle into the local excitation point nodule of the left medial vastus muscle fascia in rats,twice a week,for 4 weeks.The mechanical foot contraction reflex threshold of the leftfoot were measured weekly in the pre/post modeling and post-intervention groups of rats.At 4 weeks after treatment,hematoxylin-eosin staining was used to observe the morphological changes in the muscle tissue of the leftmedial femoral muscle of rats,ELISA was used to detect the levels of substance P and β-endorphin in the serum and the gray matter around the midbrain aqueduct.Immunohistochemistry was used to detect positive expression of microglia markers(Iba-1)and c-fos in the gray matter around the midbrain aqueduct.Western blot assay was used to detect the expression level of brain-derived neurotrophic factor protein in the periaqueductal gray.RESULTS AND CONCLUSION:Compared with the blank group,the mechanical pain threshold of the rats in the model group and the stagnant moving needle group decreased after modeling(P<0.05).After 4 weeks of treatment,the mechanical pain threshold of the rats in the stagnant moving needle group was higher than that in the model group(P<0.05).Hematoxylin-eosin staining results showed that in the model group,the muscle fibers of the leftlower limb medial femoral muscle of rats were disorganized,unequal in thickness,myocytes were enlarged,with inward movement of the nucleus,rounded contracture nodules and tension bands;whereas in the stagnant moving needle group,the muscle fibers were arranged in a neat way,the myocytes were angular,and the contracture nodules were occasionally seen.Compared with the blank group,the expression of substance P in the serum of the model group was significantly higher(P<0.05),while the levels of β-endorphin in serum and substance P and β-endorphin in brain were decreased(P<0.01).Compared with the model group,the level of serum substance P in the stagnant moving needle group was decreased(P<0.05),and the levels of serum β-endorphin and brain substance P and β-endorphin were increased(P<0.05).Compared with the blank group,the positive expression of c-fos and Iba-1 and the protein of brain-derived neurotrophic factor in the model group were increased(P<0.05).Compared with the model group,the positive expression of c-fos in the stagnant moving needle group was increased(P<0.05),and the positive expression of Iba-1 and the protein of brain-derived neurotrophic factor were decreased(P<0.05).These findings suggest that stagnant moving needle may indirectly promote the release of β-endorphin by microglia polarized to the M2 phenotype and increase the excitability of c-fos neurons by inhibiting the activity of microglia in the gray matter around the periaqueductal gray and downregulating the expression of brain-derived neurotrophic factor protein,thereby reducing the degree of central sensitization and effectively relieving myofascial pain syndrome.

Objective:To evaluate the effect of cathepsin B(CTSB)/NOD-like receptor pyrin domain containing 3(NLRP3)pathway on the polarization of macrophages induced by LPS.Methods:The well-growing RAW264.7 mouse mononuclear macrophage lines were cultured in vitro and divided into 3 groups(n=6)according to the random number table method:control group(C group),LPS group(L group)and LPS+CA074-me(CTSB inhibitors)group(B group).C group was cultured normally for 24 h,L group was cultured with LPS concentration of 1 μg/ml medium for 24 h.B group was pretreated with CTSB inhibitor CA074-me 30 μmol/L for 1 h before LPS induction,and co-cultured with LPS concentration of 1 μg/ml medium for 24 h.After 24 hours,the morphological changes of the cells were observed by microscope,the concentrations of IL-1β and IL-18 in the supernatant were determined by ELISA.The ex-pressions of cathepsin B precursor(pro-CTSB),mature cathepsin B(mature-CTSB),NLRP3,apoptosis-related speck protein(ASC)and apoptosis-related speck protein-1(caspase-1)were detected by Western blot.The mRNA expression levels of CD32,inducible ni-tric oxide synthase(iNOS),arginase 1(Arg-1)and CD206 were detected by qRT-PCR.The positive expression rates of M1 macro-phage surface marker CD86 and M2 macrophage surface marker CD206 were detected by flow cytometry.Results:Compared with group C,the morphology of cells in groups L and B became larger and pseudopodia appeared.The concentrations of IL-1β and IL-18 in cell supernatant were increased,the expressions of pro-CTSB,mature-CTSB,NLRP3,ASC and caspase-1 were increased,and the expressions of CD32,iNOS mRNA were up-regulated and the positive rates of CD86 and CD206 were increased(P<0.01).Arg-1 and CD206 mRNA in group B were up-regulated(P<0.01).Compared with group L,the pseudopodia of group B were reduced,and the morphology was closer to group C.The concentration of IL-1β and IL-18 in the supernatant,the expression of mature-CTSB,NLRP3,ASC and caspase-1,CD32 and iNOS mRNA and the positive rate of CD86 were down-regulated in group B.The expression of pro-CTSB,Arg-1 and CD206 mRNA and the positive rate of CD206 were increased(P<0.01).Conclusion:Inhibition of CTSB/NLRP3 pathway can reduce the inflammatory response,reduce the LPS-induced polarization of RAW264.7 cells to M1 macrophages,and pro-mote their polarization to M2 macrophages.

Objective:To Explore the changes of annexin A2 and vascular endothelial cadherin (VE-Cad) in patients with cerebral infarction (CI) undergoing emergency thrombolysis, and analyze their relationship with progression within 10 d.Methods:Using a prospective research method, 78 patients with CI were selected from October 2019 to June 2022 in Xi'an International Medical Center Hospital, and all patients were treated with emergency thrombolysis. The serum levels of annexin A2 and VE-Cad before and after thrombolysis were measured by enzyme-linked immunosorbent assay, and the National Institute stroke scale (NIHSS) was used to assess patients' neurologic impairment. The baseline data, imaging findings at admission and routine laboratory examination indexes were recorded. The progression within 10 d after thrombolysis was recorded. Pearson correlation analysis was used to analyze the correlation between annexin A2, VE-Cad and NIHSS score. Multivariate Logistic regression was used to analyze the independent risk factors of progression within 10 d after thrombolysis in patients with CI. The value of annexin A2 and VE-Cad in predicting the progression within 10 d after thrombolysis in patients with CI was evaluated by the receiver operating characteristics (ROC) curve. A restricted cubic spline model was drawn to evaluate the dose-response relationship between annexin A2, VE-Cad and the progression within 10 d after thrombolysis in patients with CI.Results:Compared with before thrombolysis, the annexin A2 after thrombolysis was significantly higher: (24.50 ± 3.27) μg/L vs. (20.86 ± 3.84) μg/L, the VE-Cad and NIHSS score were significantly lower: (4.72 ± 1.05) mg/L vs. (6.81 ± 1.31) mg/L and (8.64 ± 2.35) scores vs. (13.01 ± 2.86) scores, and there were statistical differences ( P<0.01). Before and after thrombolysis, Pearson correlation analysis result showed there was a negative correlation between annexin A2 and NIHSS score ( r =-0.796 and - 0.568, P<0.01), and a positive correlation between VE-Cad and NIHSS score ( r = 0.820 and 0.502, P<0.01). Among 78 patients with CI treated with emergency thrombolysis, 7 cases (8.97%) experienced progression within 10 d. There were statistical differences in hypertension, diabetes, hyperlipidemia, onset to thrombolysis time, infarct site, systolic blood pressure, triacylglycerol, high-density lipoprotein cholesterol, and the NIHSS score, annexin A2, VE-Cad before and after thrombolysis between patients with progression within 10 d after thrombolysis and patients without progression within 10 d after thrombolysis ( P<0.05 or <0.01); there were no statistical differences in gender composition, age, body mass index, coronary heart disease, atrial fibrillation, smoking, alcohol consumption, family history of stroke, carotid plaques, blood glucose, diastolic blood pressure, white blood cell count, platelet count, total cholesterol low-density lipoprotein cholesterol between the two groups ( P>0.05). After adjusting for hypertension, diabetes and hyperlipidemia, multivariate Logistic regression analysis result showed that the infarction site, onset to thrombolysis time, VE-Cad after thrombolysis and annexin A2 after thrombolysis were still independent factors of progression within 10 d after thrombolysis in patients with CI ( OR = 2.570, 2.496, 3.147 and 0.352; 95% CI 1.285 to 5.139, 1.303 to 4.781, 1.629 to 6.080 and 0.158 to 0.782; P<0.05 or <0.01). ROC curve analysis results showed that the area under the curve of annexin A2 combined with VE-Cad after thrombolysis to predict the progression within 10 d after thrombolysis in patients with CI was significantly larger than that of annexin A2 and VE-Cad after thrombolysis alone (0.898 vs. 0.822 and 0.799, χ2 = 2.17 and 1.98, P = 0.039 and 0.048). The optimal cutoff values of annexin A2 and VE-Cad after thrombolysis were <23.27 μg/L and >4.92 mg/L, with a sensitivity of 88.24%, and a specificity of 77.05%. The restricted cubic spline analysis result showed that the continuous changes in annexin A2 after thrombolysis were roughly negatively correlated with the progression within 10 d after thrombolysis in patients with CI ( OR = 0.720, 95% CI 0.561 to 0.930, P = 0.010), the continuous changes in VE-Cad after thrombolysis were roughly positively correlated with the progression within 10 d after thrombolysis in patients with CI ( OR = 1.450, 95% CI 1.126 to 1.188, P = 0.004). When annexin A2<23.80 ng/L and VE-Cad>5.25 mg/L after thrombolysis, the risk of progression within 10 d after thrombolysis in patients with CI significantly increased. Conclusions:The expression of annexin A2 increases and VE-Cad decreases after emergency thrombolysis in patients with CI, and the expression levels of both are closely related to the degree of neurologic impairment, and the risk of progression within 10 d after thrombolysis could be determined clinically by detecting their changes.

Objective:To evaluate the long-term clinical efficacy and the influence on anal function of surgery combined with ustekinumab (UST) in active Crohn's disease (CD) patients with perianal fistula.Methods:A retrospective cohort study was conducted. Clinical data of active CD patients with perianal fistula undergoing surgery combined with UST at Longhua Hospital of Shanghai University of Traditional Chinese Medicine from August 2020 to December 2022 were collected. The primary endpoints were clinical healing rate, Wexner score, and anorectal manometry values at week 52 of treatment. Secondary endpoints included the Crohn's disease activity index (CDAI), perianal Crohn's disease activity index (PDAI), laboratory indicators [C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), fecal calprotectin (FC) ], endoscopic remission rate, Van Assche score, and radiographic healing rate at week 52 of treatment.Results:A total of 28 patients were included, with 22 males (78.6%) and 6 females (21.4%) ; age (25.2 ± 7.7) (16.0-52.0) years. There was 1 patient (3.6%) of simple anal fistula. There were 27 patients (96.4%) of complex anal fistulas, including 12 of high intersphincteric type, 15 of high transsphincteric type, in which 15 with branched tracts (or ≥2 fistula tracts). The pre-treatment CDAI was 187.0 (156.0, 245.0), and the PDAI was 10.0 (9.0, 12.0). Among the 28 patients, 23 (82.1%) underwent fistulotomy, 1 (3.6%) underwent transanal opening of intersphincteric space (TROPIS), and 4 (14.3%) underwent video-assisted anal fistula treatment (VAAFT) combined with fistula-tract laser closure (FiLaC). All the patients received UST treatment postoperatively, without concurrent use of immunosuppressants or corticosteroid therapy. At week 52 of treatment, 28 (100%) patients achieved clinical healing. Compared to pre-treatment, Wexner score of patients at week 52 of treatment was significantly lower [0 (0, 0) vs. 1.0 (0, 3.0), P < 0.001], maximum anal sphincter pressure increased [ (137.6±40.9) mmHg vs. (105.1±29.2) mmHg, P < 0.001], maximum anal sphincter contraction time extended [9.0 (5.0, 15.0) s vs. 4.0 (2.0, 6.0) s, P < 0.001], and there was no significant decrease in anal resting pressure ( P > 0.05). Compared to pre-treatment, CDAI, PDAI, Van Assche scores, and simple endoscopic score for Crohn's disease (SES-CD) of patients at week 52 of treatment all significantly decreased (all P < 0.001), and CRP, ESR, and FC all decreased (all P < 0.05), with statistically significant differences. The radiographic healing rate at week 52 of treatment was 75.0% (21/28), and the radiographic remission rate was 92.9% (26/28). The endoscopic remission rate was 57.1% (16/28), and the endoscopic response rate was 82.1% (23/28) . Conclusion:The long-term clinical healing rate of active CD patients with perianal fistula receiving surgery combined with UST is high, and the anal function can be improved significantly.