OBJECTIVE To investigate the effect and underlying mechanism of hydroxytyrosol(HT)on mouse chondrocyte injury induced by oxidative stress.METHODS Mouse chondrocytes were incubated with varying concentrations of HT 0-400 μmol L-1 for 24 h,and the viability of the mouse chondrocytes was assessed using CCK-8 kit.An oxidative stress model of chondrocytes was estab-lished by the addition of H2O2 200 μmol L-1.The experimental groups included the cell control group,H2O2 group,and H2O2+HT 10,50 and 250 μmol·L-1 groups.After 24 h,the mRNA expression levels of interleukin-6(IL-6),cyclooxygenase-2(COX-2),prostaglandin E2(PGE2),inducible nitric oxide synthase(iNOS),matrix metalloproteinase-3(MMP-3),MMP-13,a disintegrin and metalloproteinase with throm-bospondin motifs-4(ADAMTS-4),ADAMTS-5,SRY-box transcription factor-9(SOX-9)and aggrecan(ACAN)in mouse chondrocytes were detected by real-time quantitative PCR,the intracellular reactive oxygen species(ROS)level in chondrocytes was measured with 2,7-dichlorodihydrofluorescein diace-tate(DCFH-DA)staining,while the mitochondrial membrane potential was evaluated using JC-1 staining.After 48 h,the protein expression levels of iNOS,COX-2,MMP-13,and type Ⅱ collagen(Col-2)in mouse chondrocytes were detected using Western blotting.RESULTS HT at concentrations≤350 μmol·L-1 had no significant effect on the survival of mouse chondrocytes.Compared with the cell control group,after 24 h,the mRNA expression levels of IL-6,COX-2,PGE2,iNOS,MMP-3,MMP-13,ADAMTS-4 and ADAMTS-5 in the chondrocytes of mice in the H2O2 group were increased,while the mRNA expression levels of SOX-9 and ACAN were decreased.Additionally,there was an elevation in the ROS level and a significant loss of mitochondrial membrane potential in the chondrocytes of mice.Compared with the H2O2 group,after treatment with HT 10,50 and 250 μmol·L-1,there were significant decreases in mRNA expression levels of IL-6,COX-2,PGE2,iNOS,MM P-3,MMP-13,ADAMTS-4 and ADAMTS-5,the mRNA expressions of SOX-9 and ACAN were increased,the ROS level was lowered.After treatment with HT 50 and 250 μmol L-1,the loss of mitochondrial membrane potential was ameliorated.Compared to the cell control group,the protein expressions of iNOS,COX-2 and MMP-13 were upregulated in the H2O2 group,while the protein expression of Col-2 was downregulated after 48 h.Compared to the H2O2 group,treatment with HT at concentrations of 10,50 and 250 μmol·L-1 resulted in decreased protein expressions of iNOS,COX-2 and MMP-13 in mouse chondrocytes,but the protein expression of Col-2 increased following treatment with HT 50 and 250 μmol L-1.CONCLUSION HT can ameliorate H2O2-induced chondrocyte injury by reducing intracellular ROS levels and alleviating the loss of mitochondrial membrane potential,suppressing the release of inflammatory cytokines,inhibiting catabolic processes,and promoting anabolic activities.

Objective:To investigate the safety and efficacy of trans-breast endoscopic approach for lateral neck dissection (LND) in the treatment of thyroid cancer.Methods:A retrospective analysis was conducted on 80 patients who underwent trans-breast endoscopic LND at the Second Affiliated Hospital of Zhejiang University School of Medicine from Dec. 2023 to Aug. 2024. Demographic characteristics, operative duration, postoperative complications, and pathological results were analyzed, with data presented using descriptive statistics.Results:The cohort included 7 males and 73 females, with a mean age of (30.0 ± 6.7) years. The mean diameter of thyroid cancer lesion was (1.3 ± 0.7) cm. The average operative duration was (4.6 ± 1.5) hours, with a 0.0% (0/80) rate of conversion to open surgery. Intraoperative recurrent laryngeal nerve (RLN) "loss of signal" occurred in 6.3% (5/80) of cases. The mean hospital stay was (2.3 ± 1.0) days. Postoperative complications included transient hoarseness in 3.8% (3/80) and transient hypoparathyroidism in 10.0% (8/80), with no cases of permanent hoarseness or hypoparathyroidism. The mean total number of dissected lymph nodes was 47.8 ± 18.6, with 8.3 ± 6.2 metastatic lymph nodes. The incidence of postoperative lymphatic leakage was 3.8% (3/80), and no cases of incision infection were observed. The mean visual analog scale (VAS) score for pain was 4.2 ± 0.4. The mean follow-up duration was 10.3 months, at 3-month follow-up, the cosmetic satisfaction score was 4.5 ± 0.4.Conclusions:The trans-breast endoscopic approach for LND is safe and effective, with high patient cosmetic satisfaction. With appropriate patient selection, it can be considered a viable surgical option for lateral neck dissection in the treatment of thyroid cancer.

Objective:To investigate the safety and efficacy of trans-breast endoscopic approach for lateral neck dissection (LND) in the treatment of thyroid cancer.Methods:A retrospective analysis was conducted on 80 patients who underwent trans-breast endoscopic LND at the Second Affiliated Hospital of Zhejiang University School of Medicine from Dec. 2023 to Aug. 2024. Demographic characteristics, operative duration, postoperative complications, and pathological results were analyzed, with data presented using descriptive statistics.Results:The cohort included 7 males and 73 females, with a mean age of (30.0 ± 6.7) years. The mean diameter of thyroid cancer lesion was (1.3 ± 0.7) cm. The average operative duration was (4.6 ± 1.5) hours, with a 0.0% (0/80) rate of conversion to open surgery. Intraoperative recurrent laryngeal nerve (RLN) "loss of signal" occurred in 6.3% (5/80) of cases. The mean hospital stay was (2.3 ± 1.0) days. Postoperative complications included transient hoarseness in 3.8% (3/80) and transient hypoparathyroidism in 10.0% (8/80), with no cases of permanent hoarseness or hypoparathyroidism. The mean total number of dissected lymph nodes was 47.8 ± 18.6, with 8.3 ± 6.2 metastatic lymph nodes. The incidence of postoperative lymphatic leakage was 3.8% (3/80), and no cases of incision infection were observed. The mean visual analog scale (VAS) score for pain was 4.2 ± 0.4. The mean follow-up duration was 10.3 months, at 3-month follow-up, the cosmetic satisfaction score was 4.5 ± 0.4.Conclusions:The trans-breast endoscopic approach for LND is safe and effective, with high patient cosmetic satisfaction. With appropriate patient selection, it can be considered a viable surgical option for lateral neck dissection in the treatment of thyroid cancer.

OBJECTIVE To investigate the effect and underlying mechanism of hydroxytyrosol(HT)on mouse chondrocyte injury induced by oxidative stress.METHODS Mouse chondrocytes were incubated with varying concentrations of HT 0-400 μmol L-1 for 24 h,and the viability of the mouse chondrocytes was assessed using CCK-8 kit.An oxidative stress model of chondrocytes was estab-lished by the addition of H2O2 200 μmol L-1.The experimental groups included the cell control group,H2O2 group,and H2O2+HT 10,50 and 250 μmol·L-1 groups.After 24 h,the mRNA expression levels of interleukin-6(IL-6),cyclooxygenase-2(COX-2),prostaglandin E2(PGE2),inducible nitric oxide synthase(iNOS),matrix metalloproteinase-3(MMP-3),MMP-13,a disintegrin and metalloproteinase with throm-bospondin motifs-4(ADAMTS-4),ADAMTS-5,SRY-box transcription factor-9(SOX-9)and aggrecan(ACAN)in mouse chondrocytes were detected by real-time quantitative PCR,the intracellular reactive oxygen species(ROS)level in chondrocytes was measured with 2,7-dichlorodihydrofluorescein diace-tate(DCFH-DA)staining,while the mitochondrial membrane potential was evaluated using JC-1 staining.After 48 h,the protein expression levels of iNOS,COX-2,MMP-13,and type Ⅱ collagen(Col-2)in mouse chondrocytes were detected using Western blotting.RESULTS HT at concentrations≤350 μmol·L-1 had no significant effect on the survival of mouse chondrocytes.Compared with the cell control group,after 24 h,the mRNA expression levels of IL-6,COX-2,PGE2,iNOS,MMP-3,MMP-13,ADAMTS-4 and ADAMTS-5 in the chondrocytes of mice in the H2O2 group were increased,while the mRNA expression levels of SOX-9 and ACAN were decreased.Additionally,there was an elevation in the ROS level and a significant loss of mitochondrial membrane potential in the chondrocytes of mice.Compared with the H2O2 group,after treatment with HT 10,50 and 250 μmol·L-1,there were significant decreases in mRNA expression levels of IL-6,COX-2,PGE2,iNOS,MM P-3,MMP-13,ADAMTS-4 and ADAMTS-5,the mRNA expressions of SOX-9 and ACAN were increased,the ROS level was lowered.After treatment with HT 50 and 250 μmol L-1,the loss of mitochondrial membrane potential was ameliorated.Compared to the cell control group,the protein expressions of iNOS,COX-2 and MMP-13 were upregulated in the H2O2 group,while the protein expression of Col-2 was downregulated after 48 h.Compared to the H2O2 group,treatment with HT at concentrations of 10,50 and 250 μmol·L-1 resulted in decreased protein expressions of iNOS,COX-2 and MMP-13 in mouse chondrocytes,but the protein expression of Col-2 increased following treatment with HT 50 and 250 μmol L-1.CONCLUSION HT can ameliorate H2O2-induced chondrocyte injury by reducing intracellular ROS levels and alleviating the loss of mitochondrial membrane potential,suppressing the release of inflammatory cytokines,inhibiting catabolic processes,and promoting anabolic activities.

Objective:To explore the feasibility of anastomosis between bladder and intestine of experimental rabbit by drag anastomosis.Methods:In this study 40 Japanese big-eared rabbits were randomly divided into two groups through random number table, the experimental group and the control group, each group with 20 rabbits. In the experimental group, the bladder neck was fixed to the catheter and then the catheter was drawn outward. With the traction of the catheter, the bladder neck was anastomosed with the distal intestinal tube by means of suture free. The control group was anastomosed by regular interrupted suture of bladder and intestine. The operation time, anastomosis time, intraoperative blood loss, postoperative urinary leakage rate and postoperative anastomotic healing of rabbits in the two groups were compared.Results:The operation time of the experimental group was shorter than that of the traditional interrupted suture anastomosis group [(33.26±2.79)min vs. (35.25±1.83)min, P=0.014]. The anastomosis time of the experimental group was significantly shorter than that of the traditional interrupted suture anastomosis group[(7.55±1.2)min vs. (8.65±1.03 min), P=0.005]. The intraoperative blood loss in the experimental group was similar to that in the control group[(6.47±2.41) ml vs. (6.75±1.83) ml, P=0.691]. The event of contrast media extravasation occurred in 2 of the 10 experimental rabbits after receiving cystography in the experimental group, and the urinary leakage rate was 20%(2/10). In the control group, contrast media extravasation occurred in 1 of the 9 experimental rabbits after receiving cystography, and the urinary leakage rate was 11.1%(1/9), and the difference of the two groups was not statistically significant ( P=0.348). Anastomotic healing score was (2.0±0.7) in the experimental group, and (2.1±0.74) in the control group ( P=0.767). Conclusions:The bladder-intestine drag-and-bond anastomosis technique, with significantly shorter anastomosis time, was feasible, easy and convenient. Our research provides an experimental and theoretical basis for the clinical application of drag-and-bond anastomosis technique in clinic.

Objective:To compare the efficacy of staged versus elective operation for treating acute closed fracture-dislocation of tarsometatarsal joint complex.Methods:A retrospective cohort study was used to analyze the clinical data of 26 patients with acute closed fracture-dislocation of tarsometatarsal joint complex admitted to Tongji Hospital of Tongji University from January 2017 to January 2021, of whom 18 were males and 8 were females, aged 32-52 years [(44.3±5.2)years]. According to the time from injury to admission, 14 patients admitted within 8 hours after injury underwent staged surgical treatment (staged group), and 12 patients admitted more than 8 hours after injury underwent elective surgery (elective group). In the staged group, emergency reduction and temporary internal fixation with K-wire were done under the supervision of a C-arm X-ray machine in the first stage, while after the swelling subsided, open reduction and internal fixation were done for tarsometatarsal joint fracture-dislocation in the second stage. In the elective group, open reduction and internal fixation were performed for tarsometatarsal joint fracture-dislocation on a scheduled basis after the swelling subsided. The operation time, hospitalization time and fracture healing time were recorded. The visual analogue score (VAS) and American Orthopedic Foot and Ankle Society (AOFAS) midfoot score were evaluated before operation, at 1, 6, 12 months after operation and at the final follow-up. The rate of complications was observed after operation.Results:All patients were followed up for 12-24 months [(18.5±3.8)months]. The operation time, hospitalization time and fracture healing time in the staged group were (77.3±5.6)minutes, (14.3±2.2)days and (12.3±1.2)weeks, respectively, significantly shorter than those in the elective group [(101.5±7.5)minutes, (20.3±5.2)days and (14.3±2.2)weeks] (all P<0.01). VAS significantly decreased and AOFAS midfoot score significantly increased in both groups as postoperative time increased (all P<0.05). There were no significant differences in VAS between the two groups before operation, at 12 months after operation or at the final follow-up (all P>0.05). The VAS at 1, 6 months after operation was (4.4±0.8)points and (2.1±0.4)points in the staged group, significantly lower than those in the elective group [(6.0±1.0)points and (3.5±0.6)points] (all P<0.01). There was no significant difference in preoperative AOFAS midfoot score between the two groups ( P>0.05). The AOFAS midfoot score at 1, 6, 12 months after operation and at the final follow-up was (67.6±4.5)points, (75.7±5.2)points, (83.6±2.2)points and (85.9±4.3)points in the staged group, significantly higher than those in the elective group [(60.2±3.9)points, (70.2±3.4)points, (75.4±3.3)points and (78.7±4.4)points] (all P<0.01). The rate of complications was 14.3% (2/14) in the staged group, significantly lower than that in the elective group [33.3% (4/12)] ( P<0.05). Conclusion:Compared to traditional elective surgery, staged surgery for acute closed fracture-dislocation of tarsometatarsal joint complex has the advantages of shortened operation time, hospitalization time and fracture healing time, eary pain relief, improved functional recovery of the foot and reduced postoperative complications.

Objective:To analyze the correlation between the posterior malleolus fracture and fixation and the rotational stability of the ankle and to explore the surgical indications for posterior malleolus fracture aiming to provide the theories for the diagnosis and treatment of disorder.Methods:Twenty fresh frozen cadaver specimens were selected. Further, the extent of the tibial insertion of the posterior inferior tibiofibular ligament (PITFL) and inferior transverse tibiofibular ligament (ITTFL) complex was dissected and measured. Based on the tibial insertion of the ligament complex, the model for the supination-external rotation degree 3 ankle fracture with a posterior malleolar fragment and syndesmosis diastasis was created. Moreover, the area threshold of the posterior tibial insertion of posterior malleolus fracture was biomechanically assessed. The difference of the antirotating ability of the ankle-stiffness between simple posterior malleolus fixation and simple syndesmotic fixation was analyzed statistically.Results:The PITFL and ITTFL were presented in all specimens with relatively broad in PITFL tibial insertion. The PITFL was attached to the posterolateral tibia. The distance between the highest point of the tibial insertion and the articular line was 45.2±5.6 mm, while the ITTFL was attached to the posterior distal tibia. The distance between the highest point of the tibial insertion and the articular line was 5.5±1.0 mm. The width of the tibial insertion of the PITFL and ITTFL complex decreased as the distance from the joint line increased. Biomechanical analysis showed that the threshold of posterior area of posterior malleolus fracture was 1/4S. The stiffnesses of posterior malleolus fixation and syndesmosis stabilization were 0.264±0.080 N·m/° and 0.164 ± 0.061 N·m/°, respectively. The percentage of stiffness restored by posterior ankle fixation was 60.9%±10.2%, which was greater than that by syndesmosis stabilization 37.5%±7.9% ( t=17.09, P<0.001) . Conclusion:The surgical technique for posterior malleolus fracture should consider restoration of the axial and rotational stability of the ankle simultaneously. Posterior malleolus fracture fixation is recommended when the syndesmosis is unstable with the area ratio of posterior tibial insertion of posterior malleolus fracture greater than or equal to 1/4. Syndesmotic fixation is proposed to restore and maintain the rotational stability of the ankle when the syndesmosis is unstable with the area ratio less than 1/4. Regardless of the area ratio, the surgical indications for stable syndesmosis depend on the impact of the posterior malleolus fracture on the axial stability of tibiotalar joint, on the involved articular surface area and on the displacement degree of posterior malleolus fragment.

Objective:To propose a new classification of posterior malleolus fracture to further clarify its pathoanatomy.Methods:Twenty fresh frozen cadaver specimens of normal morphology of lower limbs were selected and dissected and the extent of the tibial insertion of posterior malleolus associated ligaments was measured. At the same time, a retrospective case series analysis was made on the clinical and CT information of 296 patients with posterior malleolus fracture treated at Tongji Hospital of Tongji University from January 2012 to July 2020 or at Karamay Central Hospital from January 2018 to July 2020. The percentage of articular involvement of the fracture, proximal displacement of the posterior malleolus fracture and extent of posterior talar subluxation were measured. A clinically practical new classification system for posterior malleolus fracture was created by correlating posterior malleolus associated ligaments with CT images of posterior malleolus fracture. The new classification included three types: type I was posterior malleolus fracture with only the tibial insertion of inferior transverse tibiofibular ligament involved; type II was posterior malleolus fracture with both the tibial insertions of inferior transverse tibiofibular ligament and posterior inferior tibiofibular ligament involved, which was divided into subtypes IIA and IIB based on the presence of articular cartilage and subchondral bone damage, compression or Die-Punch fragments; type III was posterior malleolus fracture that involved all the tibial insertions of inferior transverse tibiofibular ligament, posterior inferior tibiofibular ligament and posterior tibiotalar ligament, which was sub-classified into subtypes III A and III B according to number of fracture fragments. Anatomic characteristics of the extent of the tibial insertion of posterior malleolus associated ligament, CT imaging parameters for posterior malleolus fracture and corresponding fracture typing were determined. In addition, the new classification system for posterior malleolus fracture was compared with Haraguci classification and Mason classification.Results:Posterior malleolus associated ligaments included the posterior inferior tibiofibular ligament, inferior transverse tibiofibular ligament and posterior tibiotalar ligament from posterolateral to posteromedial tibia. The posterior inferior tibiofibular ligament was attached to the posterolateral tibia and the distance between the highest point of its tibial insertion and the joint line was (45.2±5.6)mm. The inferior transverse tibiofibular ligament was attached to the posterior distal tibia and the distance between the highest point of its tibial insertion and the joint line was (5.5±1.0)mm. The posterior tibiotalar ligament was attached to the posterior colliculus and intercollicular groove of the medial malleolus and the distance between the center of its tibial insertion and the intercollicular groove was (2.5±0.6)mm. Among 296 patients with posterior malleolus fracture, there were 36 patients with type I, 229 with type II (150 type IIA, 79 type IIB) and 31 with type III (11 type IIIA, 20 type IIIB). The percentage of articular involvement of the fracture, proximal displacement of posterior malleolus fracture and extent of posterior talar subluxation in type IIB fracture were significantly greater than those in type II A fracture [23.7(18.6, 28.8)% vs. 18.4(12.7, 21.7)%, 4.1(2.1, 6.0)mm vs. 1.9(0.2, 3.0)mm, 4.7(1.5, 6.2)mm vs. 2.3(1.1, 3.0)mm] (all P<0.01). The proximal displacement of posterior malleolus fracture and extent of posterior talar subluxation in type III fracture were significantly greater than those in type II fracture [7.2(6.0, 8.2)mm vs. 2.7(0.4, 4.0)mm, 10.1(6.0, 15.0)mm vs. 3.1(1.1, 5.0)mm] (all P<0.01). The new classification for posterior malleolus fracture combined the posterior malleolus ligament and injury mechanism of posterior malleolus fracture as compared with Haraguchi classification, which not only further detailed the classification, but also was more practical in clinic for increased the severity of injury was elevated with higher classification level. The new classification was more comprehensive as compared with Mason classification for it mainly added the type of simple-rotation-type posterior malleolus fracture (type IIA of the new classification). Conclusions:In combination with posterior malleolus associated ligaments, injury mechanism and fracture morphology, posterior malleolus fracture is divided into three types. The new classification system more comprehensively describes pathoanatomy of posterior malleolus fracture that contributes to related basic research and clinical diagnosis and treatment.

Objective:To develop an efficient and rapid method for the isolation and cultivation of human scalp dermal papilla cells from small specimens.Methods:Hair-bearing skin specimens measuring 0.5 cm × 0.5 cm -0.5 cm × 1 cm in size were obtained from the scalp of 3 patients with pigmented nevus and 6 with sebaceous nevus during surgery in Department of Dermatology, the First Hospital Affiliated to Army Medical University from September 2018 to January 2019. The subcutaneous fat layer containing hair follicles was cut out of the specimens, and hair follicles were sorted with ophthalmic forceps, which were subsequently digested with 0.6% dispase Ⅱ for 30 minutes, then with 0.2% collagenase Ⅳ at 37 ℃ for 30 - 60 minutes, and were centrifuged to obtain hair papillae. Morphological observation was performed on the isolated hair papillae, and dermal papilla cells were cultured, passaged and identified.Results:Under the microscope, the hair papillae isolated by two-step enzyme digestion of small scalp specimens were intact, and showed an inverted pear-like shape, and residual dermal sheaths could be observed around some hair papillae. However, no hair papilla was isolated by one-step enzyme digestion. With the two-step enzyme digestion method, the hair papilla separation rate was 60.8% ± 2.1%, the adherence rate of the dermal papilla cells at 72 hours was 86.6% ± 3.9%, the time for cells to emigrate out of hair papillae was 0.5 - 3.0 days, the total operation duration was 2.0 - 3.0 hours, and the actual operation duration after subtraction of digestion duration was 1.0 - 1.5 hours. The dermal papilla cells isolated by the two-step enzyme digestion method could grow in an aggregative pattern in early stage, but grew in a non-aggregative pattern after 8 passages.Conclusion:The two-step enzyme digestion of small specimens is a simple and efficient method for isolating human scalp dermal papilla cells.

Objective: To evaluate the predictive accuracy of fine needle aspiration (FNA) and BRAF V600E mutation in distinguishing papillary thyroid carcinoma and other thyroid nodules. Methods: This retrospective cohort study included 93 patients with papillary thyroid carcinoma who treated at Department of Thyroid Surgery, the Second Affiliated Hospital of Zhejiang University, College of Medicine from September 2016 to May 2018. There were 21 males and 72 females with age of (43.2±11.3) years (range: 19 to 67 years). All the patients got the examinations of FNA and BRAF V600E mutation by Amplification Refractory Mutation System, and subsequently underwent thyroid surgeries. The results of cytopathology, frozen section and pathology were collected and analyzed. The predictive accuracy of FNA cytology and BRAF V600E mutation was calculated. Results: In the 93 collected cases, 91 were diagnosed as papillary thyroid carcinoma postoperation, and the accurate predictive rate was 97.8%. Subgroup analysis was performed according to Bethesda System, the predictive rates were: unsatisfactory (Ⅰ) 6/6, benign (Ⅱ) 0/0, atypia of undetermined significance or follicular lesion of undetermined significance (Ⅲ) 16/17, follicular neoplasm or suspicious for follicular neoplasm (Ⅳ) 97.2% (35/36), suspicious for malignancy (Ⅴ) 100% (28/28), and malignant (Ⅵ) 6/6, respectively. Conclusion Thyroid nodules with BRAF V600E mutation can be strongly speculated as papillary thyroid carcinoma.