ObjectiveOptimize the latex perfusion technique and apply it to the construction of a murine craniofacial venous vascular model.Methods A total of nine 8-week-old male C57BL/6 mice weighing (25.0±1.3) g were randomly divided into three groups: 60% latex physiological saline group, 60% latex heparin group, and 30% latex heparin group. After completion of the perfusion, the specimens were immersed in 4 °C formalin fixative for 24 h, followed by dissection, observation, and measurement of the extracranial blood vessel diameters. Results After 200 μL latex perfusion solution was injected into the external jugular vein, the supraorbital vein, infraorbital vein, temporal vein, retrofacial vein, masseter vein and external jugular vein were perfused in each group.After comparing the perfusion degree of the distal branches of blood vessels, sublingual vein and tip venule, it was found that the 30% latex heparin group had the best perfusion effect, followed by the 60% latex heparin group, and the 60% latex saline group had the worst perfusion effect.ConclusionThe optimized latex perfusion technique can effectively infuse the veins in the head and face of mice, and this technique can provide a good reference for the study of the direction and morphology of facial veins in mice.

Obiective:To investigate the design, the key points of surgery and the outcome of short-term follow-up of hemi-elbow-arthroplastic bionic total humeral prosthesis by using 3D-print.Methods:The clinical data of 8 patients with humerus malignant tumor who underwent total humerus resection and a 3D-printed auxiliary hemi-elbow-arthroplastic total humeral prosthesis replacement from March 2018 to March 2020 at Peking University People's Hospital were retrospectively analyzed. The operative data, oncological outcome, complication profiles of prosthesis and functional status of all 8 patients were also analyzed. There were 3 males and 5 females with a mean age of 5-37 years. Histological diagnosis included 5 cases of osteosarcoma, 2 cases of Ewing's sarcoma and 1 case of chondrosarcoma.Results:The time for production of the prosthesis was (9.8±2.7) d, the operation time was (209±23) min and intraoperative hemorrhage was (569±173) ml. All the prostheses were implanted successfully and no patients experienced intraoperative complications. While 2 patients had postoperative complications, 1 case of temporary palsy of radial nerve and 1 case of local recurrence. LARS artificial ligament or hernia patch was used to reconstruct joint capsule and tendon-ligament attached around the elbow and shoulder joint. The flexion and extension of the elbow was (118±15)° (100-140°) and (11±9)° (0-25°), and the abduction and anteflexion of the shoulder was (28±12)° (15-50°) and (26±9)° (15-40°), respectively. The postoperative Musculoskeletal Tumor Society (MSTS)-93 scale score was (24.1±1.5) scores. The median follow-up time was 17 months (12-32 months), 7 patients had disease-free survival and 1 patient survived with tumor.Conclusions:The novel 3D-printed total humeral prosthesis with hemi-elbow-arthroplasty has a good perioperative safety, which is effective in restoring the function of elbow joint, solving the problem of stress concentration of ulnar marrow lever of total elbow joint prosthesis and lowering long-term wear rate and loosening rate of prosthesis.

Objective:We designed and applied a novel, bionic megaprosthesis for distal femur, which substituted the hinged articulation with an artificial ligament and a constrained tibial insert in order to maintain articular stability and reduce stress and the risks of wearing, loosening and breakage.Methods:We reviewed the cases of distal femoral osteosarcoma in children population who were initially treated in our center during 2019. Twelve cases of bionic distal femoral prosthesis (bionic group) and 21 cases of fixed-hinge distal femoral prosthesis (control group) were included. In the bionic group, there were six female and six male with the mean age as 8.8±2.6 y. One patient had pathological fracture. Staging according to Enneking system included 1 case of stage IIA, 10 cases of stage IIB and 1 case of stage III. In the control group, there were seven female and fourteen male with the mean age as 8.6±2.2 y. Pathological fracture was seen in three cases. Staging included 15 cases of stage IIB and 6 cases of stage III.We investigated the peri-operative safety, oncological outcome, complication profiles and post-operative functional status of the bionic prosthesis by comparing the baseline data, operative data, and oncological and prosthetic outcomes between the two groups.Results:The two groups were comparable in terms of baseline data including gender, age, height, weight and onset duration, and operative data including duration of operation (126.7±27.9 min vs 143.3±38.9 min, P=0.203), intra-operative hemorrhage (162.5±212.3 ml vs 247.6±175.6 ml, P=0.224) and duration of wound drainage (6.3±2.4 d vs 6.4±3.4 d, P=0.908). The mean follow-up duration of bionic group was 16.0±4.7 months, during which time three patients had systemic progression and one of them died of disease. Another patient had local recurrence and subluxation of the prosthesis that was treated by amputation. Subluxation might be related to the huge tumor mass and excessive resection of the quadriceps femoris. As for the control group, the mean follow-up duration was 12.7±4.5 months. Three patients had systemic progression and all patients were still alive by last follow-up. Local recurrence was seen in two patients and was treated by excision. No prosthetic complications were seen in the control group. The averaged range of motion (ROM) was greater in the bionic group than that in the control group (120.6°±13.6° vs 92.0°±7.7°, P<0.05), but the MSTS 93 scores were similar between groups (29.1±0.9 vs 29.5±0.6, P=0.337). Conclusion:Compared with the fixed-hinge distal femoral prosthesis, the bionic prosthesis had good peri-operative safety, did not increase the risks of prosthetic complications, and could achieve a better range of motion and a similar functional status based on the results of short-term follow-up.

Objective:To investigate the causes,prevention and treatment of femoral artery puncture related complications caused by the application of resuscitative endovascular balloon occlusion of the aorta (REBOA) in the resection of pelvic and sacral tumors.Methods:Clinical data of 23 patients with femoral artery puncture related complications who received REBOA in the resection of pelvic and sacral tumors from August 2010 to August 2018 at the Musculoskeletal Tumor Center,Peking University People′s Hospital were retrospectively analyzed.There were 8 males and 15 females,with the age of (37.0±16.2) years (range:15 to 65 years).Arterial access via the Seldinger technique for REBOA was obtained in the right common femoral artery of 18 cases,and in the left of 6 cases.An arterial sheath with a diameter of 11 to 12 F(1 F≈0.33 mm) was used for the patient.The occurrence and treatment of postoperative complications were analyzed.Results:Acute femoral arterial thrombosis occurred in 18 patients,which was managed by open repair 48 hours postoperatively.Among the 349 patients admitted before 2015 who received hemostasis by compression after femoral artery sheath removal,12 patients (3.4%) developed acute femoral artery thrombosis.While the 476 patients admitted after 2015 who used a percutaneous suture device to close the femoral artery wound,6 patients (1.3%) developed acute femoral artery thrombosis.One case of retroperitoneal hematoma and 1 case of femoral pseudoaneurysm were found and surgically fixed.Postoperative follow-up was (40±18) months (range:13 to 108 months).Three cases with chronic lower extremity ischemia were confirmed by Doppler ultrasonography during 1 to 5 years follow-up.Two of them had minimal symptoms and denied further treatment,while the other one received femoral-femoral artery bypass surgery to restore distal flow for pain and numbness relief.Conclusions:Acute femoral arterial thrombosis is the most common femoral artery puncture.Technique refinement of REBOA,the use of percutaneous suture device and close follow-up can reduce the approach-specific complications,and help to detect and treat the complications timely,which may popularize the clinical application of REBOA.

Objective:To investigate the causes,prevention and treatment of femoral artery puncture related complications caused by the application of resuscitative endovascular balloon occlusion of the aorta (REBOA) in the resection of pelvic and sacral tumors.Methods:Clinical data of 23 patients with femoral artery puncture related complications who received REBOA in the resection of pelvic and sacral tumors from August 2010 to August 2018 at the Musculoskeletal Tumor Center,Peking University People′s Hospital were retrospectively analyzed.There were 8 males and 15 females,with the age of (37.0±16.2) years (range:15 to 65 years).Arterial access via the Seldinger technique for REBOA was obtained in the right common femoral artery of 18 cases,and in the left of 6 cases.An arterial sheath with a diameter of 11 to 12 F(1 F≈0.33 mm) was used for the patient.The occurrence and treatment of postoperative complications were analyzed.Results:Acute femoral arterial thrombosis occurred in 18 patients,which was managed by open repair 48 hours postoperatively.Among the 349 patients admitted before 2015 who received hemostasis by compression after femoral artery sheath removal,12 patients (3.4%) developed acute femoral artery thrombosis.While the 476 patients admitted after 2015 who used a percutaneous suture device to close the femoral artery wound,6 patients (1.3%) developed acute femoral artery thrombosis.One case of retroperitoneal hematoma and 1 case of femoral pseudoaneurysm were found and surgically fixed.Postoperative follow-up was (40±18) months (range:13 to 108 months).Three cases with chronic lower extremity ischemia were confirmed by Doppler ultrasonography during 1 to 5 years follow-up.Two of them had minimal symptoms and denied further treatment,while the other one received femoral-femoral artery bypass surgery to restore distal flow for pain and numbness relief.Conclusions:Acute femoral arterial thrombosis is the most common femoral artery puncture.Technique refinement of REBOA,the use of percutaneous suture device and close follow-up can reduce the approach-specific complications,and help to detect and treat the complications timely,which may popularize the clinical application of REBOA.

Objective:To investigate the effect of miR-23b on the malignant phenotype and the sensitivity of lenvatinib in human hepatocellular carcinoma cells.Methods:Human hepatocellular carcinoma cell line HepG2, SMMC-7721 and QGY-7703 were transfected with miR-23b mimic and its control, respectively. CCK-8 and EdU assay were used to detect cell proliferation. Transwell assay were used to detect changes in cell migration and invasion. Tube formation assay were used to detect vasculogenic mimicry formation. The comparison of the mean between groups was analyzed by t-test.Results:CCK-8 results showed that the A values ??of human hepatocellular carcinoma cell line HepG2 and SMMC-7721 in the miR-23b mimic group were 0.325 ± 0.011 and 0.537 ± 0.026, respectively, which were significantly lower than the control group 0.430±0.017 and 0.752 ± 0.051 ( P < 0.05). Transwell assay result showed that the number of cell migration of human hepatocellular carcinoma cell line HepG2 and SMMC-7721 in the miR-23b mimic group was (517.220 ± 32.873) and (242.327 ± 20.793), respectively, which were significantly lower than that of the control group (724.130 ± 15.142) and (424.432 ± 27.212) ( P < 0.01). Simultaneously, the number of cell invasions in the miR-23b mimic group were (55.671 ± 7.514) and (64.670 ± 6.011), respectively, which were significantly lower than those in the control group (124.320 ± 11.782) and (156.204 ± 12.501) ( P < 0.01). Tube formation assay showed that the number of tube forming branches of hepatocellular carcinoma cell line QGY-7703 and SMMC-7721 in the miR-23b mimic group was (489.824 ± 42.035) and (435.201 ± 44.143), respectively, which were significantly lower than that of the control group (878.620 ± 31.618) and (785.430 ± 38.723) ( P ??< 0.01). In addition, EdU results showed that after miR-23b combined with lenvatinib, the positive rates of EdU staining of hepatocellular carcinoma cell line HepG2 and SMMC-7721 in the miR-23b mimic group were (32.905 ± 1.342)% and (24.811 ± 0.820)%, respectively, which were significantly lower than the control group (52.623 ± 2.441)% and (38.702 ± 1.312)% ( P < 0.05). Conclusion:miR-23b can inhibit the proliferation, migration, invasion and vasculogenic mimicry formation, and enhance the sensitivity of lenvatinib drug in human hepatocellular carcinoma cells.

Objective To investigate the proliferative inhibition effect and mechanism of total saponins from marsdenia tenacissima on human liver cancer HepG2 cells. Methods Human liver cancer HpeG2 cells cultured conventionally were divided into the marsdenia tenacissima saponins A control group, the experimental group and the blank control group. The experimental group was treated with different mass concentrations of total saponins from marsdenia tenacissima (0.14, 0.29, 0.58, 1.15, 2.13 mg/ml), while the marsdenia tenacissima saponins A control group was treated with marsdenia tenacissima saponins A (1.0 mg/ml), and the blank control group was established stimultaneously. The methyl thiazolyl tetrazolium (MTT) method was used to detect the effect of total saponins from marsdenia tenacissima on the activity of HepG2 cells, and the cell morphology was observed by using inverted microscopy. The cell apoptosis rate was detected by using flow cytometry from Annexin V-FITC/PI staining. The expressions of bcl-2, bax and p53 were analyzed by using Western blot after the drug effect. Results The results of cell activity test showed that total saponins from marsdenia tenacissima could inhibit the proliferation of HepG2 cells at the concentration of 0.29, 0.58, 1.15, 2.13 mg/ml, which was positively correlated with the concentration. The half maximal inhibitory concentration (IC 50) of cell growth inhibition of total saponins from marsdenia tenacissima on HepG2 cells was 0.75 mg/ml. Under inverted microscopy, the adherent cells were significantly reduced, the cells fell off into clusters and the debris was increased after the effect of total saponins from marsdenia tenacissima. Moreover, flow cytometry showed that total saponins from marsdenia tenacissima could increase the late apoptosis rate of HepG2 cells (P< 0.01). Western blot showed that the expression of bcl-2 was 0.62±0.16, 0.31±0.15, 0.84±0.09 and 1.00±0.11 respectively in the experimental group (total saponins from marsdenia tenacissima 0.58 mg/ml and 2.13 mg/ml), the marsdenia tenacissima saponins A control group and the blank control group;the expression of bax protein was 0.75±0.10, 0.83±0.12, 1.00±0.14 and 0.15±0.02, respectively in the above four groups; the expression of p53 protein was 0.63±0.08, 0.78±0.11, 1.00±0.13 and 0.18±0.02 respectively in the above four groups. The protein expression of bcl-2 was decreased and the protein expressions of bax and p53 were increased in the experiment group, and there was a statistical difference between the experiment group and the blank control group (P< 0.05). Conclusions Total saponins from marsdenia tenacissima can upregulate the expression of bax by upregulating the expression of p53 gene, and inhibit the expression of bcl-2, which would cause the cascade reaction to induce the apoptosis of HepG2 cells. Its inhibitory effect can be realized through mitochondrial pathmay to induce apoptosis.

Objective To evaluate the safety and efficacy of denosumab in treatment of patients with pelvic giant cell tumor of bone (GCTB) during perioperative period. Methods This is a retrospective observational study. Twenty-three patients diagnosed with pelvic GCTB undergoing perioperative denosumab treatment in Musculoskeletal Tumor Center of Peking University People's Hospital from January 2014 to December 2016 were reviewed. The subjective adverse reactions and mandibular X-ray films were used to assess the drug safety. As for efficacy, imaging findings (including X-ray, CT, magnetic resonance imaging) were reviewed. MSTS-93 scoring system was applied in the postoperative functional assessment. Histological response rate, objective response rate, clinical benefit rate and event-free survival rate were all used to deficit the efficacy of denosumab in the treatment of pelvic GCTB combined with surgery. All the results of postoperative were compared statistically with pelvic GCTB patients who underwent surgery in the same hospital from 1999 to 2009. Results All the patients were firstly diagnosed as classic GCTB except for one case which was malignant pelvic GCTB. All patients received denosumab preoperatively and/or postoperatively, and the average number of medications was 8.43. According to the surgical patterns, patients were divided into intralesional surgery group (13 cases) and wide resection group (10 cases). The follow-up was 5-47 months(mean:27.30 months),recurrence was observed in 2 cases in the intralesional surgery group, none in the wide resection group. After drug administration, 13 cases were partial response, 7 cases were stable disease, the objective response rate was 65.0 % (13/20), and the histologically clearance rate of giant cells was 85.0 % (17/20). No case of osteonecrosis of the jaw was observed in this study, and all laboratory indicators were normal. The average postoperative MSTS-93 score was 26.87. Compared with pelvic GCTB patients who underwent surgical treatment from 1999 to 2009, in the intralesional surgery group, there was no significant difference in the recurrence rate [15.4 % (2/13) vs. 30.8 % (4/13), P = 0.514], but the limb function was significantly increased (P= 0.002). Conclusions Denosumab combined with surgery plays an important role in the multidisciplinary treatment of pelvic GCTB. The neoadjuvant strategy can reduce patient's intraoperative blood loss by shrinking the tumor size which makes the intralesional curettage surgery possible, and also diminishing the recurrence rate. But more attention should be paid to secondary malignant GCTB during the use of denousmab.

Objective To explore the inhibitory effect of curcumin on LPS-induced inflammation and the activation of Toll-like receptor 4 (TLR4 )/NADPH oxidase/reactive oxygen species (ROS)signaling pathway in vascular smooth muscle cells (VSMCs).Methods Primary VSMCs were cultured and divided into control group, LPS group,LPS + curcumin 5 μmol/L group,LPS + curcumin 10 μmol/L group and LPS + curcumin 30 μmol/L group.Cell activity was observed by MTT assay.The secretion of tumor necrosis factor-α(TNF-α)and interleukin-1 (IL-1)was measured by enzyme linked immunosorbent assay (ELISA)kits.The mRNA expressions of TLR4 and p22phox were detected by real-time PCR.Expression of intracellular ROS was measured by flow cytometry. Results The activities of VSMCs were not significantly affected by curcumin at the concentration between 0 and 80 μmol/L.Curcumin (5,10 and 30 μmol/L)significantly inhibited LPS-induced oversecretion of TNF-αand IL-1, as well as overexpression of TLR4 and p22phox at the mRNA and protein levels,and ROS production in VSMCs in a concentration-dependent manner.Conclusion Curcumin has a concentration-dependent inhibitory effect on the secretion of inflammatory cytokine,overexpressions of TLR4 and p22phox,and production of ROS in VSMCs stimulated by LPS.Furthermore,curcumin may partly depend on TLR4/NADPH oxidase/ROS signaling pathways to inhibit inflammation in LPS-induced VSMCs.

Objective To evaluate the feasibility and clinical effect of Argon plasma coagulator in simple enucleation for small renal cell carcinoma. Methods On the basis of successful performing the animal experience of coagulating therapy on the wound tissue during partial nephrectomy with Argon plasma coagulator in rabbit models, 10 cases of simple enucleation for small renal cell carcinoma with Argon plasma coagulator were accomplished. Results Both with the standard of stopping bleeding of wound tissue by Argon plasma coagulator and with the standard of re-spraying the wound tissue for 2 s after stopping bleeding using Argon plasma coagulator, the depth of wound tissue necrosis without blocking the renal pedicle is deeper than that with blocking the renal pedicle(P=0. 012 and P=0. 002, respectively).If the wound tissue was re-sprayed for 2 s after stopping bleeding by Argon plasma coagulator, the depth of the wound tissue necrosis without blocking the renal pedicle was deeper than that just with blocking the renal pedicle(P=0. 007 and P=0. 002,respectively). In the part of application in clinical, all procedures were successfully completed. The mean operative time was 163 min (range, 100-210 min) and mean blood loss was 230 ml (range, 100-400 ml). Drainage tube was pulled out 1 month after operation in 1 case for being allergic to absorbable hemostatic gauze, and the mean pulling drainage tube out time in others was 4. 2 d (range, 3-5 d). During a mean follow-up of 22 months (range, 10-38 months), no local tumor recurrence and distant metastasis was found. Conclusion Argon plasma coagulator can be used in simple enucleation for small renal cell carcinoma, and the clinical effectiveness is ideal.