BACKGROUND:Heterotopic ossification of skeletal muscle is a clinically serious complication.For heterotopic ossification of skeletal muscles,the cells involved in the process of heterotopic ossification remain unclear. OBJECTIVE:To investigate the involvement of myocytes,fascia cells,and endothelial cells in the process of heterotopic ossification in skeletal muscle and to observe the cell origin of heterotopic ossification in skeletal muscle induced by bone morphogenetic protein 4. METHODS:Both C2C12 cells and the myotubes formed by the C2C12 cells in the induction medium were cultured,and 500 ng/mL bone morphogenetic protein 4 was added to the medium respectively,and whether the C2C12 cells and myotubes continued to proliferate within 10 days under the treatment were observed under a microscope.Myogenic cells(L6,derived from rats)and fibroblast-derived cells(derived from human)were co-cultured.After treatment with 500 ng/mL bone morphogenetic protein 4 and 10 ng/mL transforming growth factor-β,osteogenic and chondrogenic differentiation potential within 21 days were observed using Safranine O staining and Alcian blue staining.Using transgenic animal FVB/N-TgN(TIE2-LacZ)182Sato mice,15 μL of adeno-associated virus-bone morphogenetic protein 4(5×1010 PFU/mL)were implanted in the thigh muscle space of genetic mice for 10 and 14 days.X-gal staining was used to observe the formation of new blood vessel endothelium in the differentiated bone. RESULTS AND CONCLUSION:(1)Bone morphogenetic protein 4 caused myotube breakdown and increased C2C12 cell proliferation.Compared with other groups,the pure fibroblast-derived cell group had a higher area of positive alcian blue and safarin O staining(P<0.05)and a lower area of alkaline phosphatase staining(P<0.05),while the pure L6 group had a bigger area of alkaline phosphatase staining(P<0.05)but a smaller area of positive alcian blue and safarin O staining(P<0.05).(2)Transplantation of adeno-associated virus-bone morphogenetic protein 4-adsorbed gelatin sponge into FVB/N-TgN(TIE2-LacZ)182Sato mice resulted in heterotopic ossification.(3)X-gal staining results demonstrated that there was no obvious staining in chondrocytes and differentiated bones and Tie2+ endothelial cells did not participate in the formation of the alienated bone.(4)These findings verify that fibroblasts are the primary source of osteoblasts during the adeno-associated virus-bone morphogenetic protein 4-induced ectopic endochondral ossification in skeletal muscle,but myogenic cells are the main source of osteoblasts.Tie2+ endothelial cells might not be the cell source for cartilage and bone.

Objective Exploring the mechanism of electroacupuncture's anti anxiety effect through methods such as hippocampal metabolomics and 16S rRNA sequencing.Methods Rats were randomly divided into normal group,model group,electroacupuncture group,and diazepam group.A rat model of anxiety disorder was prepared using chronic restraint stress method.During the modeling period of the electroacupuncture group,simultaneous"Zusanli"acupoint electroacupuncture intervention was performed for 30 minutes each time,once a day,for 21 days.The diazepam group received daily gavage of diazepam for a total of 21 days.After the completion of modeling,open field experiments and elevated cross maze experiments were conducted to observe the behavior of rats.Subsequently,pathological staining was performed to observe the pathological changes in the hippocampus and colon of rats.Metabolomics detection of changes in metabolites in the hippocampus of rats.16S rRNA and short chain fatty acid analysis were used to detect changes in gut microbiota and short chain fatty acids in rats.Western blot detection of Occludin in the colon and TLR4,NF-κB P65 and NLRP3 in the hippocampus of rats.Immunofluorescence detection of IBA-1,NLRP3,and IL-1β in the hippocampus Protein expression.Result The results of the open field experiment showed that compared with the normal group,the total activity distance(P<0.001),central area activity distance(P<0.01),and average velocity(P<0.01)of the model group decreased.Compared with the model group,the total activity distance(P<0.05,P<0.001),central area activity distance(P<0.05,P<0.01),and average velocity(P<0.05,P<0.01)of the electroacupuncture group and diazepam group all increased.The results of the elevated cross maze experiment showed that compared with the normal group,the model group had a decrease in OE%(P<0.001)and OT%(P<0.001).Compared with the model group,both the electroacupuncture group and the diazepam group could increase OE%(P<0.05,P<0.001)and OT%(P<0.01,P<0.001).The HE staining results of the hippocampus showed that the CA1 area of the hippocampus in the normal group rats showed clear and hierarchical structures in various regions of the hippocampus.In the hippocampus of the model group,a small amount of neural cell nuclei in the CA1 area were observed to be wrinkled and deeply stained,with unclear cell boundaries and irregular arrangement,and the cytoplasm was vacuolate.The intervention of electroacupuncture and diazepam can alleviate the above pathological phenomena to varying degrees,respectively;The results of hippocampal metabolomics showed that electroacupuncture can improve hippocampal metabolic disorders caused by modeling,mainly involving taurine and low taurine metabolism,cysteine and methionine metabolism,and regulation of glycine,serine,and threonine metabolic pathways;The results of 16S rRNA sequencing and short chain fatty acid detection showed that electroacupuncture can improve the disturbance of intestinal microbiota caused by modeling,and can regulate serum LPS and levels of butyric acid,caproic acid,and valeric acid.HE staining and Western blot results showed that the model group had pathological damage to the intestine,and compared with the normal group,the expression of Occludin in the colon of the model group decreased(P<0.05).However,the intervention of electroacupuncture and diazepam could improve the pathological damage of the colon and upregulate the expression of Occludin(P<0.05,P<0.05).The immunofluorescence results showed that compared with the normal group,the expression levels of IBA-1,NLRP3,and IL-1β in the hippocampus of the model group increased(P<0.05,P<0.01,P<0.01),while compared with the model group,the electroacupuncture group and diazepam group could reduce the expression levels of IBA-1(P<0.05,P<0.05),NLRP3(P<0.05,P<0.05),and IL-1β(P<0.05,P<0.05).Western blot results showed that compared with the normal group,the expression levels of TLR4,NF-κB P65,and NLRP3 proteins in the hippocampus of the model group increased(P<0.05,P<0.001,P<0.05).Compared with the model group,the electroacupuncture group and diazepam group were able to downregulate the expression levels of NF-κB P65(P<0.01,P<0.001)and NLRP3(P<0.05,P<0.05)proteins.Conclusion The anxiolytic of electroacupuncture involves the regulation of the"microbiota-gut-brain"axis.

【Objective】 To analyze the risk factors of ileus after radical cystectomy, and to construct a nomogram predictive model accordingly. 【Methods】 Clinical data of patients who underwent radical cystectomy during Jan.2018 and Dec.2021 were collected. The risk factors related to postoperative ileus were assessed with Logistic univariate and multivariate regression analysis. After that, the predictive model was constructed and its specificity and accuracy were verified. 【Results】 A total of 326 patients were included, 65 of whom developed ileus. Statistical analysis showed that gender, lymph node dissection, serum creatinine and albumin were correlated with postoperative ileus. The area under the receiver operating characteristic curve of the model was 0.769 (95%CI:0.724-0.802). Bootstrap correction curve showed that the model had good prediction accuracy. 【Conclusion】 Male, lymph node dissection, elevated postoperative serum creatinine and postoperative blood albumin decrease are predictors of ileus. The nomogram predictive model based on these predictors can predict the probability of ileus after radical cystectomy.

Objective:To summarize and analyze the current application status of oral mucosal graft (OMG) technique in the repair of ureteral strictures in China, and clarify the feasibility, safety and effectiveness of this technique.Methods:The 175 patients who underwent repair of ureteral stricture using oral mucosal patches from June 2015 to February 2022 were etrospectively analyzed in 14 medical centers in China, including 49 cases in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 32 cases in Affiliated Seventh Medical Center of PLA General Hospital, 3 cases in The Second Hospital of Anhui Medical University, 6 cases in The First Affiliated Hospital of Zhengzhou University, 56 cases in Peking University First Hospital, 3 cases in Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 3 cases in Shanghai Sixth People' s Hospital, 4 cases in General Hospital of Estern Theater Command, 4 cases in Lanzhou University Second Hospital, 2 cases in Guizhou Province People 's Hospital, 2 cases in Peking University People' s Hospital, 5 cases in Jinzhou First People's Hospital, 5 cases in The First Affiliated Hospital of Wannan Medical College, 1 case in Shandong Provincial Hospital. In this study, 127 patients (72.6%) used lingual mucosal patches, 32(18.3%) labial mucosa, and 16(9.1%) buccal mucosa. The surgical approach for OMG ureteral reconstruction was mainly minimally invasive, with robot-assisted laparoscopy in 84 patients (48.0%), traditional laparoscopic surgery in 87 patients (49.7%), and open surgery in only 4 patients (2.3%). There were 133 males and 42 females with an average age of (35.0±17.2) years. The mean body mass index (BMI) and stenosis length were (23.1±4.1) kg/m 2 and (4.7±1.8) cm, respectively. The stricture was located in the left ureter in 116 patients, right ureter in 58 case and bilateral ureter in 1 case. The most common causes of ureteral stricture were endoscopic surgery in 88(50.3%)patients, congenital stricture in 55(31.4%)patients, failed ureteroplasty in 29(16.6%)patients, history of extracorporeal shock wave lithotripsy in 13(7.4%)patients, radiotherapy history in 3(1.7%)patients and other causes in 6(3.4%)patients. Strictures were mainly located in the upper ureter, accounting for 61.7% (108/175 cases), followed by 36.0% (63/175) at the ureteropelvic junction and 2.3%(4/175)in the middle ureter. According to the surgical methods, the patients were divided into robot-assisted laparoscopic surgery group ( n=84), traditional laparoscopic surgery group ( n=87)and open surgery group ( n=4). Subgroup analysis of patients in robot-assisted laparoscopic and traditional laparoscopic surgery groups was performed. There were no significant difference in preoperative data between the two groups except for age (32.0±18.3) years vs.(37.0±15.9)years, P=0.040], BMI[(22.5±4.3)kg/m 2 vs. (23.7±3.6)kg/m 2, P=0.028], and etiology of stenosis [endoscopic injury, 34(40.5%) vs. 53(60.9%), P=0.012]. Preoperative hydronephrosis and stricture length were assessed by CTU and ureterography. Ureterography 7-9 weeks after surgery showed patency of the reconstructed segment, or no recurrence of hydronephrosis was judged as success. Evaluate the operation method, operation time, success rate, length of OMG in repairing ureteral stricture between laparoscopic and robot-assisted groups. Results:The overall success rate of oral mucosal graft repair surgery reached 97.7%(171/175). The success rate of ureteral reconstruction in the two groups were 96.4%(81/84)and 98.9%(86/87), respectively ( P=0.351), and the difference was not statistically significant. There was no significant difference for operation time, intraoperative blood loss, and mean oral mucosal length between the robotic and laparoscopic groups[(244.7±85.8) min and (222.7±83.5)min ( P=0.116), (58.9±38.6) ml and (68.4±45.5) ml ( P=0.217), (5.0±2.0) cm and (4.6±1.5) cm ( P=0.350)], respectively.Postoperative complications were reported in 23 (13.1%) patients, such as fever, urinary leakage, lymphatic leakage, infection, but only 2 (1.4%) cases patients had complications of Clavien-Dindo score ≥ Ⅲ. The two patients developed urinary stricture after surgery with failed conservative treatment, and no urinary stricture occurred following endoscopic treatment.The short-term (three months after surgery)incidence of complications in the site where the oral mucosa was taken, such as difficulty in opening mouth, pain, and swelling, was 12.0% (21/175), and there was no significant difference for oral complications between patients harvesting different length of mucosal graft. Conclusions:Ureteroplasty with oral mucosal graft is a safe, feasible and reliable technique for ureteral reconstruction. At present, minimally invasive technology is the main surgical approach for ureteroplasty, and there is no significant difference in operation time and success rate between robotic surgery and laparoscopic surgery.

Objective:To construct a C57BL/6 mouse model of simulating transurethral thulium laser vaporization prostatectomy.Methods:Twelve male C57BL/6 mice were selected to undergo transvesical vaporization resection of the urothelium covering the urethra of the prostate using thulium laser. The urethral tissue of the prostate was retrieved on the 1st, 3rd, 5th, and 7th days after the surgery. HE staining was used to observe the process of re-epithelialization of the urethral wound of the prostate. Immunohistochemical (IHC) staining was used to detect whether the re-epithelialized cells of the urethral wound of the prostate expressed urothelin Ⅲ (UPⅢ).Results:On the first day after surgery, HE staining showed complete destruction to the urothelium covering the urethra of the prostate, with a large amount of coagulative necrotic tissue on the wound surface, and IHC staining showed no expression of UPⅢ on the wound surface. On the 3rd day after surgery, HE staining showed that there were still no regenerated epithelial cells on the wound surface, with coagulation necrosis tissue significantly reduced, and the urethral cavity was clearly visible. And IHC staining showed no expression of UPⅢ on the wound surface. On the 5th day after surgery, HE staining showed 1-2 layers of regenerated epithelial cells lacking cell polarity on the wound surface, and IHC staining showed that the regenerated epithelial cells expressed UPⅢ. On the 7th day after surgery, HE staining showed 4-6 layers of polar regenerated epithelial cells on the wound surface, and IHC staining showed the multiple layers of regenerated epithelial cells expressing UPⅢ.Conclusions:Based on the simulation of transurethral thulium laser vaporization resection of the prostate, the thulium laser and ultra micro endoscope system were used to vaporize the urothelium covering the urethra of the prostate, and the process of urethral re-epithelialization of the prostate can be observed after surgery. The establishment of the C57BL/6 mouse model simulating thulium laser vaporization prostatectomy provides a new research platform for studying the mechanism of wound repair after prostatectomy.

Objective:To evaluate the application of amide proton transfer weighted (APTw) magnetic resonance imaging(MRI) for grading of brainstem glioma (BSG) in children.Methods:Twenty-five children (16 males and 9 females) aged 0.7-12.4(5.6±3.3)years were diagnosed as BSG by surgery or biopsy in Beijing Children′s Hospital from December 2019 to March 2022, including 13 cases of low-grade BSG and 12 cases of high-grade BSG. APTw imaging and conventional MRI were performed on a 3.0 T MRI scanner. The differences in gender distribution, age, conventional MRI appearance, APTw signal intensity and apparent diffusion coefficient (ADC) between children with high and low grade BSG were analyzed. Receiver operating characteristic (ROC) curve was used to analyze the efficacy of APTw signal intensity in the differential diagnosis of high and low grade BSG, and Youden index was calculated to obtain the optimal diagnostic threshold. Pearson′s correlation coefficient analysis was used to evaluate the correlation between APTw signal intensity and Ki-67 expression.Results:There was no significant difference in age and gender distribution between high-grade and low-grade BSG patients. The maximum diameter of high-grade BSG was significantly larger than that of low-grade BSG [(4.7±0.9) vs. (3.1±1.7)cm; t=-2.94, P=0.007]; the maximum signal intensity of APTw (APTw max) in high-grade BSG was significantly higher than that in low-grade BSG [(4.9±0.6)% vs. (3.0±1.2)%; t=-5.14, P<0.001]; the average signal intensity of APTw (APTw mean) in high-grade BSG was significantly higher than that in low-grade BSG[(3.6±0.4%) vs. (2.7±1.1)%; t=-2.66, P=0.014].The area under the curve(AUC)of APTw max in distinguishing high-and low-grade BSG was 0.897; with 4.07% as the optimal diagnostic threshold of APTw max, the sensitivity for the diagnosis of high-grade BSG was 0.917 and specificity was 0.846. The AUC of APTw mean in distinguishing high-and low-grade BSG was 0.769; with 2.85% as the optimal diagnostic threshold of APTw mean, the sensitivity for the diagnosis of high-grade BSG was 0.917 and specificity was 0.692. There was a positive correlation between the value of APTw max and Ki-67 expression( r=0.453, P=0.023). Conclusion:APTw imaging is helpful to distinguish high-grade and low-grade BSG in children. APTw max value can be used to effectively evaluate the proliferative activity of BSG in children.

Objective:To investigate the clinical treatment of large segmental humeral defects with unilateral external fixation and bone transport.Methods:A retrospective study was conducted of the 9 patients who had been treated at Department of Orthopedics, Shenzhen People's Hospital for large segmental humeral defects from September 2017 to June 2019. They were 5 males and 4 females with an average age of 29 years (from 21 to 41 years). Their defects were caused by trauma in 2 cases, by chronic osteomyelitis in 6 cases and by bone tumor in one case. The length of bone defect ranged from 4.2 to 9.0 cm, with an average of 5.9 cm. A unilateral external fixator was placed in operation, and adjusted regularly 7 to 10 days after operation for bone transport and bone lengthening to restore the length of humerus gradually. The external fixation bracket was removed after 3 to 4 layers of cortex were observed on X-ray films. Recorded were length and rate of humeral lengthening, fracture healing time, time for carrying external fixator and complications; the Disabilities of the Arm, Shoulder and Hand (DASH) scores were compared between preoperation and 15 months postoperation.Results:All the patients were followed up for 15 to 36 months (mean, 19 months). The length of lengthening averaged 5.9 cm (from 4.2 to 9.0 cm) with an average lengthening rate of 26%, the healing index 31 d/cm, the bone healing time 8.3 months, and the time for carrying external fixator 10.8 months(from 8.0 to 13.5 months). Their average DASH scores improved significantly from 25.0 ± 2.4 preoperation to 12.0 ± 1.8 at 15 months postoperation ( P<0.05). Good correction of large humeral defects was achieved in all but one case who reported temporary radial nerve paralysis. There were no such complications as neurovascular injury. The shoulder and elbow functions were basically normal after operation. Conclusions:In the treatment of large segmental humeral defects, unilateral external fixation plus bone transport can quickly repair the defects and recover the upper limb function of the patients.

Objective To demonstrate the clinical and imaging features of extrarenal malignant rhabdoid tumor (MRT) outside the central nervous system(CNS) in children and to raise awareness of the disease.Methods A retrospective database review was made of 15 patients diagnosed with extrarenal MRT outside the CNS in Beijing Children's Hospital,Capital Medical University from April 2008 to February 2017.The ultrasound,CT and magnetic resonance imaging(MRI) examinations were performed in 12,10 and 7 cases,respectively,and the clinical and imaging features were analyzed.Results The 15 patients included 8 boys and 7 girls.The age at presentation varied from 11 days to 12 years and 9 months old with a median age of 4 years old.The primary tumors were predominant solid masses,most of which were irregular,with a length of 2.20-11.70 cm [(5.87 ± 2.57) cm].The primary tumors had variable locations,relatively tended to occur in the head and neck region (6 cases) and the mediastinum (4 cases).On ultrasound,11 cases of the tumors were heterogeneous,7 cases of the tumors were accompanied with cystic change,and the solid part was mainly hypoechoic.On CT,density of 6 cases of the tumors was uneven,3 cases presented cystic change,4 cases were with calcification,and the solid part had slightly low density,with enhancement of different degrees.On MRI,7 cases of tumors showed mixed signal,5 cases with cystic change of different degrees;the solid part mainly showed isointensity or slightly hypointensity on T1 weighted image and hyperintensity or slightly hyperintensity on T2 weighted image,with restricted diffusion and predominantly heterogeneous enhancement.Medical imaging showed definite hemorrhage within tumors in 5 patients,and metastases occurred in 6 patients at the time of diagnosis,the adjacent bone destruction occurred in 3 patients,and the tumor tissue of 3 patients extended into the spinal canal.Conclusion Extrarenal MRT outside the CNS is more common in preschool kids,most of which are large and heterogeneous solid masses,often with cystic change,sometimes with hemorrhage and calcification;the solid part with restricted diffusion and inhomogeneous enhancement.Some of the patients had metastasis and local invasion.

Objective To investigate the diagnostic value of MRI in reversible splenial lesion syndrome (RESLES) in children.Methods The clinical and MRI imaging data of 14 cases of RESLES in children were retrospectively reviewed.There were 4 males and 10 females,aging 11 to 35 months.Average age was (20±3) months.MRI studies were conducted in all the cases.Two experienced doctors analyzed independently the images and reached consensus.Results Fourteen cases showed the single abnormal signal in the splenial of corpus callosum on the initial cerebral MRI.Lesions in 10 of 14 cases appeared as isointensity on T1WI,hypo-intensity in 4 cases.Lesions in 14 cases were appeared as hyper-intensity on T2WI,hyper-/slightly hyper-intensity on FLAIR T2WI.The lesions were round,oval or irregular,with fuzzy boundary.DWI showed round or oval high signal with clear boundary.No edema around the lesion and no occupying effect were detected.After treatment,8 of 14 cases were reviewed after 7-14 d,and the abnormal signal in the corpus callosum disappeared in 5 cases,while the range was significantly reduced in 3 cases.MRI were reexamined after 1-2 months in 5 cases,and the lesions disappeared in the corpus callosum.One case didn't have MRI examination again after treatment and the clinical symptoms disappeared.Conclusions The clinical manifestations of RESLES in children are lack of specificity.And the cerebral MRI imaging is characterized by the reversible solitary lesion of the splenium,which can provide a reliable basis for clinical diagnosis and prognosis.

Post competency is a kind of necessary ability for professional working.In order to train the students' excellent professional skills,we have conducted exploration and reform in post competency oriented experiment teaching,through reforming experimental courses,changing teaching methods and means of examination,constructing the laboratory etc.,so that we have built a new professional training model in pathological diagnosis and technology specialty experimental teaching.The experimental teaching reform results have improved the experimental teaching quality and the talents capability.