Marsdenia tenacissima injection, a standard Marsdenia tenacissima extract (MTE), has been approved as an adjuvant therapeutic agent for various cancers. Our previous study showed that MTE inhibited the proliferation and metastasis of prostate cancer (PCa) cells. However, the underlying mechanisms and active ingredients of MTE against PCa were not completely understood. This study revealed that MTE induced significant decreases in cell viability and clonal growth in PCa cells. In addition, MTE induced the apoptosis of DU145 cells by reducing the mitochondrial membrane potential and increasing the expression of Cleaved Caspase 3/7, Cyt c, and Bax. In vivo, DU145 xenografted NOD-SCID mice treated with MTE showed significantly decreased tumor size. TUNEL staining and Western blot confirmed the pro-apoptotic effects of MTE. Network pharmacology analysis collected 196 ingredients of MTE linked to 655 potential targets, and 709 PCa-associated targets were retrieved, from which 149 overlapped targets were screened out. Pathway enrichment analysis showed that the HIF-1, PI3K-AKT, and ErbB signaling pathways were closely related to tumor apoptosis. Western blot results confirmed that MTE increased the expression of p-AKT^Ser473 and p-GSK3β^Ser9, and decreased the expression of p-STAT3^Tyr705in vitro and in vivo. A total of 13 compounds in MTE were identified by HPLC-CAD-QTOF-MS/MS and UPLC-QTOF-MS/MS. Molecular docking analysis indicated that six compounds may interact with AKT, GSK3β, and STAT3. In conclusion, MTE induces the endogenous mitochondrial apoptosis of PCa by regulating the AKT/GSK3β/STAT3 signaling axis, resulting in inhibition of PCa growth in vitro and in vivo.
Mice ; Animals ; Male ; Humans ; Mice, Inbred NOD ; Mice, SCID ; Marsdenia ; Proto-Oncogene Proteins c-akt ; Glycogen Synthase Kinase 3 beta ; Molecular Docking Simulation ; Phosphatidylinositol 3-Kinases ; Tandem Mass Spectrometry ; Prostatic Neoplasms ; Apoptosis ; STAT3 Transcription Factor

Chronic osteomyelitis is a serious clinical problem with repeated courses and high disability rate, which seriously affects the physical and mental health of patients. Through continuous learning and summary in the process of using traditional therapies, the innovative improvements and changes had made in the treatment of osteomyelitis: Radical debridement of lesions was performed by applying the basic principles of modern bone tumor surgery. The application of microsurgical technique to transfer composite tissue flap can provide guarantee for tissue defect repair and wound closure without tension. Combined with bone grafting, local antibiotics and bone fixation, an one-stage operation has significantly improve the therapeutic effect of chronic complex osteomyelitis.

Objective:To investigate the clinical efficiency of gastrocnemius muscle flap combined with antibiotics loaded calcium sulfate in the treatment of postoperative infection and plate exposure of tibial plateau fracture in elderly patients.Methods:From January, 2015 to May, 2019, 21 elderly patients with postoperative infection and plate exposure of tibial plateau fracture were treated, including 14 males and 7 females with an average age of 72.6 years, ranging from 61 to 82 years. The average course of disease was 22.7 days, ranging from 6 to 91 days. The site of wound infection was at medial in 8 cases, lateral in 9 cases and bilateral in 4 cases. The size of wound ranged from 2.0 cm×3.5 cm to 5.0 cm×12.0 cm. All wounds were implanted with antibiotics loaded calcium sulfate and repaired by gastrocnemius muscle flap combined with skin graft after debridement. Muscle flap survival, wound healing, inflammatory index, fracture healing and knee joint function were recorded. The curative effect was evaluated by McKee infection treatment criteria and the knee joint function was evaluated by HSS scoring criteria.Results:All 21 muscle flaps survived. In 1 case, skin graft necrosis occurred in a small area, and the wound healed well after dressing change. One case had exudation which was clear and the bacterial culturing was negative, and the wound healed after 2 weeks of dressing change. The other incisions were healed in stage I, and the healing rate was 90%. All patients were followed-up for an average of 28.7(16-39) months. The redness and swelling occurred in 1 case without exudation after 2 months which disappeared after anti-infection treatment and didn't recur again. The infection recurred in 1 case after 5 months, and it which was controlled after debridement and plate removal. According to McKee criteria, 19 cases were cured, 1 improved and 1 recurred with an effective rate of 95.2%. The fracture healing time was from 3 to 7 months, with an average of 4.6 months. According to HSS scoring criteria, the knee joint function was excellent in 12 cases, good in 7 cases and moderate in 2 cases.Conclusion:After thorough debridement, gastrocnemius muscle flap combined with antibiotic loaded calcium sulfate can effectively control the infection, repair the wound, promote fracture union and restore limb function in the treatment of postoperative infection and plate exposure of tibial plateau fracture in elderly patients.

To investigate the effect and application value of transplantation of the free cutaneous fibular flap combined with antibiotic-loaded calcium sulfate artificial bone graft for the treatment of antibrachial chronic osteomyelitis of Cierny-Mader type IV. Methods From August, 2013 to May, 2017, 12 cases of ulna or (and) radius chronic osteomyelitis of Cierny-Mader type IV were treated by transplantation of the free cutaneous fibu-lar flap combined with antibiotic-loaded calcium sulfate artificial bone graft. There were 7 males and 5 females, with an average age of 36.3 (21-47) years.Pure ulnar osteomyelitis in 7 cases, radius osteomyelitis in 4 cases, and both ul-nar and radius osteomyelitis in 1 case. The average range of osteomyelitis lesions was 6.3 (3.0-9.0) cm. The area of soft tissue defect (including bone scar) ranged from 8.0 cm×2.0 cm to 15.0 cm×5.0 cm. The area of the flap was 10.0 cm×3.5cm-17.0 cm×7.0 cm.The average length of the fibular flap was 8.8 (5.0-12.0) cm.Locking plate internal fixa-tion was used in 9 cases, external fixator in 2 cases, and plate combined with external fixator in 1 case. Vancomycin/gentamicin, an effective component of calcium sulfate artificial bone, averaged 0.64 g/102.7 kU (0.4 g/64 kU-1.0 g/160 kU).Routine postoperative treatment.And monthly outpatient review in the first half year after operation, and outpatient review every 3 months after half a year.One year after operation, comprehensive evaluation of elbow, forearm and wrist function with Mayo Elbow Function Index, Anderson Forearm Double Fracture Evaluation System and Cooney Wrist Function Score. Results Vascular crisis occurred in 1 case after operation, prompt surgical exploration, and ultimately all flaps survived completely.The donor sites healed well in all cases.The lower extremity functions of donor sites had no change compared with that before operation.Followed-up of an average of 22.7 months, there were 2 cases who had sen-sory disturbance in the ulnar nerve innervation area and returned to normal 3 months after operation. The fibular flaps healed satisfactorily with an average healing time of 4.7 (3-6) months.No calcium sulphate artificial bone granules were seen on X-ray at 3 months after operation.One year after operation, bone healing, forearm appearance and wrist function recovered well, but elbow and forearm motor function recovered unsatisfactorily. Conclusion On the basis of master-ing the applied anatomy and vascular anastomosis techniques of microsurgery, this method of transplantation of the free cutaneous fibular flap combined with antibiotic-loaded calcium sulfate artificial bone graft for the treatment of an-tibrachial chronic osteomyelitis of Cierny-Mader type IV has achieved satisfactory results.The recipient area is beautiful. The bone healing is reliable.And it has little influence on the recipient area and the donor area.It is worthy of clinical application.

Objective To investigate the clinical efficacy of tissue flap combined with antibiotic-loaded calcium sulfate and autogenous iliac bone grafting at stage Ⅰ for infected nonunion of long bone.Methods A retrospective case series study was conducted to analyze the clinical data of 51 patients with infected nonunion of long bones admitted to Tongde Hospital of Zhejiang Province from January 2013 to December2017.There were 42 males and nine females,aged 19-71 years,with an average age of 36.3 years.There were 43 patients with bone nonunion at tibia,six at femur,one at humerus,and one at ulna and radius.The range of bone defect area was 1-9 cm,with an average of 2.9 cm.On the basis of thorough debridement,tissue flaps combined with antibiotic-loaded calcium sulfate and autologous iliac bone graft were performed at stage Ⅰ treatment.The survival of skin flaps,infection control,nonunion healing and complications were recorded.Johner-Wruhs joint function assessment criteria were used to evaluate limb function.Results All patients were followed up for 10-35 months,with an average of 18.3 months.All flaps survived eventually.Except for one patient with infection recurrence,the other 50 patients with infection obtained primary cure.All nonunions were healed,with an average healing time of 6.4 months ranging from 4 to 15 months.Complications included venous crisis of skin flaps in five patients,anterolateral femoral cutaneous nerve injury in 20 patients,recurrence of infection in one patient,and infection of external fixator nail tract in three patients.Function evaluation results by Johner-Wruhs criteria were excellent in 34 patients,good in 14,and moderate in three patients,with an excellent and good rate of 94％.Conclusion On the basis of thorough debridement,tissue flap combined with antibiotic-loaded calcium sulfate and autogenous iliac bone graft can control the infection effectively,promote the bone union and improve the limb function.

Objective: To summarize and explore the clinical effect and application of L-shaped flap pedicled with peroneal artery perforator in the treatment of unhealed lateral L-shaped incision after calcaneal fractures. Methods: From October 2013 to March 2015, 17 patients with unhealed L-shaped incision after calcaneal fractures were treated with one-stage thorough debridement, artificial bone filling with antibiotics-laden calcium sulphate and L-shaped flap pedicled with peroneal artery. Flap areas ranged from 8 cm × 2 cm to 11 cm × 3 cm. Donor sites are primarily sutured. Results: All 17 patients were followed up for 6 to 18 months with an average of 11.5 months. All patients with infection were cured effectively, demonstrating well survived flaps with normal elasticity and pigmentation, no scar contracture, satisfactory appearance and normal ankle joint motility. Conclusions L-shaped flap pedicled with peroneal artery perforator was an effective method in the treatment of unhealed lateral L-shaped incision after calcaneal fractures. It was suitable for coverage of the unhealed wound and worthy of being popularized.

Objective: To study the treatment of huge empyema with free vastus lateralis muscle flap. Methods: From May 2015 to December 2015, 3 cases of empyema were treated with free vastus lateralis muscle flaps, (unilateral vastus lateralis muscle flap, n=2; bilateral vastus lateralis muscle flaps with Flow-through technique, n=1). The volumes of defects were 12.4 cm×5.1 cm×6.4 cm, 11.8 cm×6.9 cm×5.6 cm, and 14.4 cm×7.6 cm×9.4 cm, respectively. Result: All flaps was survived after 23-29 months follow-up. The pus cavities were filled well. There was no empyema reoccurred. The patients felt good, without chest distress. The donor site of flap healed well. The function of lower limb was not affected. Conclusions Vastus lateralis muscle can fill the pus cavity of empyema successfully, without the sacrifice of chest shape. Free vastus lateralis muscle flap is an ideal choice to repair large empyema defection. For young patient, bilateral free vastus lateralis muscle flap can be used if the cavity is too large to fill with one flap.

Objective To analysis causes of the serious complications after the operation of the lower leg perforator pedicle screw flap, and to explore the corresponding countermeasures. Methods From June, 2012 to Au-gust, 2016, 60 cases of soft tissue defect of ankle and foot were repaired with propeller flaps pedicled with perforator of lower legs. with the area were soft tissue defect ranged from 3.0 cm ×2.0 cm to 19.0 cm ×9.0 cm, and all with bone exposure. Two cases of traumatic tissue defect, 7 cases were chronic osteomyelitis of the distal tibia, 13 cases were in-cision infection and necrosis after the operation of ankle joint fracture and Pilon fracture, 10 cases were simple inci-sion necrosis after calcaneal fracture, 18 cases were calcaneal osteomyelitis, 1 case were soft tissue defect after the ankle tumor operation, 6 cases were soft tissue necrosis after the Achilles tendon rupture, and 3 cases were soft tissue defect of the dorsum with infection. The posterior tibial artery perforator pedicled propeller flap was used in 18 cases. The pedicle of the vascular pedicle was 6.0-18.0 cm from the medial malleolus, the flap rotation was 135 °-180° . There were 42 cases of the perforator pedicle propeller flap of the peroneal artery, 5.0-18.0 cm from the pedicle of the vascular pedicle and 120°-180° rotation in the flap. The area of the flap was 9.0 cm ×3.0 cm-34.0 cm ×18.0 cm. There were 32 cases of direct suture in the donor site and 28 cases of free skin grafting. Results The color, swelling, elasticity, capillary reaction and healing of donor site were observed after operation. There was no flap ischemia occurred in 60 patients. Fourteen cases had venous reflux obstruction, all of which had swelling above II degree, 8 cases had swelling above III degree with obvious purple blood stasis, resulting in partial flap necrosis in 4 cases, all necrosis in 1 case, including 4 cases of free skin grafting, 1 case of flap transplantation and repair. There were 3 cases of necrosis after skin grafting in the flap area, all of which were partial necrosis. There was case of necrosis of the wound surface after di-rect suture of the donor site and 1 case of skin disintegration after disassembly, and all wounds healed after the replace-ment of the wound and the external use of the dried blood powder. All the 60 patients were followed-up for 12 to 30 (mean, 24.5)months. The flaps survived and the donor site scars healed well. The range of motion of the ankle was from-10°to 10°(mean, 5.6 °) and the flexion of the plantar was from 20 °to 50 °(mean, 37.8 °). Fourteen patients with venous reflux disorder were followed up for 15 to 28(mean, 22.3)months. The flap and skin graft survived well. Ankle dorsiflexion ranged from-10° to 10 °(mean, 2.4 °) and plantar flexion from 20° to 45 °(mean, 35.6 °). There was no obvious limp in walking. Conclusion Although the overall effect of the lower leg perforator pedicle propeller flap to repair the soft tissue defect of the foot and ankle is satisfactory, there are still various serious complications, which are mainly due to ia-trogenic. Doctors should strictly follow the basic principles of skin flap surgery from preoperative to postoperative, and during operation and postoperative management, so as to reduce the incidence of complications.

Objective To prepare biomimetic bone material by reconstructing type Ⅰ collagen combined with polyaspartic acid. Methods By acid hydrolysis,rat tail type Ⅰ collagen was decomposed into collagen fibers,which were then placed in the calcium phosphate mineralization solution. Under the cross-linking of glutaraldehyde,the collagen fibers were reconstructed and assembled into collagen fibers,and the calcium phosphate crystals were wrapped in the inner side of the collagen fibers for biomineralizationin. After poly aspartate acid was added,calcium hydroxyapatite calcium precursor was added into the collagen fibers to simulate thebiomimetic biomineralizationin the human body. After mineralization for 3-9 days,the bone mineralization process was observed by transmission electron microscopy and electron diffraction. Results Transmission electron microscopy and electron diffraction displayed that,after 3 days of mineralization,calcium hydroxyapatite precursor was wrapped in the collagen fiber gap,and the collagen fiber was partially mineralized. After 9 days of mineralization,calcium hydroxyapatite precursor completely infiltrated into the collagen fiber,and the amorphous calcium phosphate was transformed into hydroxyapatite calcium crystal. Thus,the simulation of bone mineralization was completed,and collagen type Ⅰ collagen/hydroxyapatite calcium biomimetic bone material was formed. Conclusion Reconstruction of type Ⅰ collagen combined with polyaspartic acid can prepare biomimetic bone material that has close chemical composition and molecular structure to the human bone tissue.

Objective To discuss the clinical effect of the phase-one treatment scheme for traumatic osteomyelitis in tibia by combining flap,vancomycin-loaded calcium sulfate and autogenous iliac bone.Methods From January,2009 to July,2014,49 patients which had traumatic osteomyelitis in tibia and met the inclusive criteria were investigated and treated.By taking these patients as treatment group A(34 cases),they were treated by adopting the phase-one treatment scheme of combing tissue flap,vancomycin-loaded calcium sulfate and autogenous iliac bone.Fifteen patients who were treated by using the phase-one treatment scheme,namely,removing the lesion,implanting vancomycin-loaded calcium sulfate and repairing the defect by means of tissue flap,were chosen as control group B.Concerning treatment group A,drainage fluid was collected after operation every day to measure the concentration of vancomycin until drainage tube was removed.All the patients were followed up to study the following indexes:the standing time of drainage tube,the healing time of fracture,infection control rate,bone nonunion rate and other complications.Results All cases were followed up during 17 to 40 months after operation and no amputation was conducted for the affected limb.To repair soft tissue defect,flap and direct suture were adopted for 25 and 9 cases respectively in group A;The results indicated that all flaps survived,the poor healing of flap defect was observed for 2 cases which were healed after dressing change.However,to repair soft tissue defect,all group B cases used flaps;results revealed that distal flap necrosis was found in 2 cases applying neurocutaneous flap,with defect exudation and infection while the 2 cases were cured after debridement and dressing change without performing a second flap operation.In group A,3 cases recurred during 5 months to 2 years after operation;in group B,it was 1;other complications included pintract infection,nonunion,numbness of anterolateral thigh,hematocele in iliac 1 region.In group B,refracture occurred for 2 cases at the original lesion location 18 and 25 months after healing and was cured after plate refixation and the graft of autogenous iliac bone;intraoperative pathology validated no recurrence of osteomyelitis.The standing time of drainage tube was (12.53±4.56) days on average for group A while (17.07±3.87) days for group B;The difference was statistically significant (P＜0.05).The healing time of fracture was (6.20±2.16) months on average for group A while(8.36±2.84) months for group B.The difference was statistically significant(P＜0.05).Conclusion In one stage treatment of localized and diffused traumatic osteomyelitis,the scheme of combining tissue flap,vancomycin-loaded calcium sulfate and autogenous iliac bone effectively shortened the healing time of fracture,increased the healing strength,and reduced the exudation after operation,without increasing infection recurrence rate.The scheme was superior to merely implanting vancomycin-loaded calcium sulfate.