Variability of BMP-2 content in DBM products derived from different long bone.

Demineralized bone matrix (DBM) has been regarded as an ideal bone substitute as a native carrier of bone morphogenetic proteins (BMPs) and other growth factors. However, the osteoinductive properties diverse in different DBM products. We speculate that the harvest origin further contributing to variability of BMPs contents in DBM products besides the process technology. In the study, the cortical bone of femur, tibia, humerus, and ulna from a signal donor were prepared and followed demineralizd into DBM products. Proteins in bone martix were extracted using guanidine-HCl and collagenase, respectively, and BMP-2 content was detected by sandwich enzyme-linked immunosorbent assay (ELISA). Variability of BMP-2 content was found in 4 different DBM products. By guanidine-HCl extraction, the average concentration in DBMs harvested from ulna, humerus, tibia, and femur were 0.613 ± 0.053, 0.848 ± 0.051, 3.293 ± 0.268, and 21.763 ± 0.344, respectively (p < 0.05), while using collagenase, the levels were 0.089 ± 0.004, 0.097 ± 0.004, 0.330 ± 0.012, and 1.562 ± 0.008, respectively (p < 0.05). In general, the content of BMP-2 in long bones of Lower limb was higher than that in long bones of upper limb, and GuHCl had remarkably superior extracted efficiency for BMP-2 compared to collagenase. The results suggest that the origin of cortical bones harvested to fabricate DBM products contribute to the variability of native BMP-2 content, while the protein extracted method only changes the measured values of BMP-2.

Repair of tendons treated with peracetic acid-ethanol and gamma irradiation by EDC combined with NHS: a morphological, biochemical and biomechanical study in vitro.

The purpose of this study was to investigate whether 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) combined with n-hydroxysuccinimide (NHS) can repair tendon damage caused by peracetic acid-ethanol and gamma irradiation sterilization. The semitendinosus tendons of 15 New Zealand white rabbits were selected as experimental materials, and the tendons were sterilized in a solution containing 1% (v/w) peracetic acid and 24% (v/w) ethanol. After 15 kGy gamma irradiation sterilization, the tendons were randomly divided into three groups (n = 10). The tendons were repaired with EDCs of 0, 2.5 and 5 mM combined with 5 mM NHS for 6 h, the tendons were temporarily stored at - 80 ± °C. The arrangement and spatial structure of collagen fibers were observed by light microscopy and scanning electron microscopy, the collagen type and collagen crimp period were observed under a polarizing microscope, and the collagen fibril diameter and its distribution were measured by transmission electron microscopy, from which the collagen fibril index and mass average diameter were calculated. The resistance of collagen to enzymolysis was detected by the free hydroxyproline test, and tensile fracture and cyclic loading tests of each group of tendons were carried out, from which the elastic modulus, maximum stress, maximum strain, strain energy density and cyclic creep strain were calculated. The obtained results showed that the gap between loose collagen fibers in the 0 mM control group was wider, the parallel arrangement of tendons in the 2.5 and 5 mM groups was more uniform and regular and the fiber space decreased, the crimp period in the 5 mM group was lower than that in the 0 mM group (P < 0.05), and the concentration of hydroxyproline in the 5 mM group (711.64 ± 77.95 µg/g) was better than that in the control group (1150.57 ± 158.75 µg/g). The elastic modulus of the 5 mM group (424.73 ± 150.96 MPa) was better than that of the 0 mM group (179.09 ± 37.14 MPa). Our results show that EDC combined with NHS can repair damaged tendons after peracetic acid-ethanol and gamma radiation treatment, and 5 mM EDC has better morphological performance, anti-enzymolysis ability and biomechanical properties than 2.5 mM EDC.

Modular intercalary prosthetic reconstruction for malignant and metastatic tumours of the proximal femur.

To illustrate the surgical technique and explore clinical outcomes of the reconstruction for the malignant and metastatic bone tumour of proximal femur with metallic modular intercalary prosthesis. Sixteen patients who underwent modular intercalary prosthetic reconstruction after tumour resection were included from April 2012 and October 2020. Prosthesis and screws parameters, resected bone length and residual bone length, clinical outcomes and survivorship were analyzed. All patients were followed up for an average of 19 months (range 1-74). In our series, 12 patients died of the progression of the primary disease at the final follow-up. The cumulative survivorship since the treatment of proximal femoral metastasis was 78.6% (11 patients) at 6 months and 38.5% (5 patients) at 1 year. The mean MSTS score was 22.25 ± 4.55 among all patients. There were no cases of loosening or breakage of the prostheses, plates or screws, despite the various measurements of prostheses and residual bones. Modular intercalary prosthetic reconstruction was an effective method for malignant tumour of the proximal femur, including the advantages of providing early pain relief, quickly restoring postoperative function, required a short operation time, and preserving the adjacent joints.

Resuscitative Endovascular Balloon Occlusion of the Aorta for Blood Control in Lumbar Spine Tumor Resection Surgery: A Technical Note.

OBJECTIVES: To describe the technique of the aorta balloon occlusion, and evaluate the blood loss in lumbar spine tumor surgery assisted by aortic balloon occlusion, and to observe the balloon-related complications. METHODS: Six patients with lumbar spine tumor underwent resuscitative endovascular balloon occlusion of the aorta prior to tumor resections in our institution between May 2018 to January 2021. Medical records including demographic, diagnosis, tumor location, surgical approach, intraoperative blood loss, surgical duration, and perioperative balloon-related complication were evaluated retrospectively. RESULTS: This series included four males and two females, with a median age of 50 years (range 22 to 69). Of these, three primary tumors were plasmacytoma, giant cell tumor of bone, and osteosarcoma, while recurrence of undifferentiated pleomorphic sarcoma (UPS), recurrence of giant cell tumor of bone (GCT), and metastatic thyroid cancer were diagnosed in cases 1, 6, and 2, respectively. L2 was involved in cases 1 and 5. L3 was involved in case 6. L4 was involved in case 2, 3, and 6. L5 was involved in case 4. One-stage total en bloc resection surgery (TES) was accomplished in all patients; of this series, signal anterior approach was conducted in case 1, signal posterior approach was utilized in cases 2, 3, and 6, while combined anterior and posterior approach was performed in cases 4 and 5. The median intraoperative blood loss was 1683 mL and ranged from 400 to 3200 mL with a median surgical duration of 442 min and a range from 210 to 810 min. During the perioperative period, no serious balloon-related complications occurred. CONCLUSIONS: Endovascular balloon occlusion of the aorta successfully controls intraoperative exsanguination, contributing to a more radical tumor resection and a low rate of tumor cell contamination in lumbar tumor surgery.

Dewatering of sewage sludge via thermal hydrolysis with ammonia-treated Fenton iron sludge as skeleton material.

In this study, the alkaline hydrothermal ferric carbon (AHFC), which was prepared by hydrothermal liquefaction method using Fenton iron sludge with NH3·H2O, was used as a skeleton materials for the dewatering of sewage sludge (SS) via thermal hydrolysis. NH2 functional group presented in the AHFC and nano-sized γ-Fe2O3 was anchored on the surface of AHFC. The NH2 functional group notably promoted thermal hydrolysis of SS from the increasing of TOC and TN value. γ-Fe2O3 showed adsorption effect to the water, resulting in decline of the dewatering rate of SS in present condition. When 100% of AHFC was added, dewatering rate of SS was decreased by 19.93% (at 160 °C), 4.50% (at 180 °C) and 8.34% (at 200 °C) respectively. 3D-EEM results showed that the degree of hydrolysis was deepened when AHFC was added for the intensity of soluble microbial products decline. AHFC promoted the decomposition of protein to form heterocycle compounds in the resulting cake according to in situ FTIR results. The nano γ-Fe2O3 catalysis to cake also can be observed for the activation energy was lower than blank in the range of 40˜60%. The study demonstrated concept and the effectiveness the reuse/recycle of the Fenton iron sludge for dewatering of SS.

Cell growth inhibition, G2/M cell cycle arrest, and apoptosis induced by chloroquine in human breast cancer cell line Bcap-37.

Chloroquine is an antimalarial drug that has been used in the treatment and prophylaxis of malaria since the 1950s. The present study was undertaken to examine the effects of chloroquine on Bcap-37 human breast cancer cells' growth, cell cycle modulation, apoptosis induction, and associated molecular alterations in vitro. The chloroquine treatment decreased the viability of Bcap-37 cells in a concentration- and time-dependent manner, which correlated with G(2)/M phase cell cycle arrest. The chloroquine-mediated cell cycle arrest was associated with a decrease in protein levels/activity of polo-like kinase 1 (Plk1), phosphorylated cell division cycle 25C (Cdc25C), phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2), phosphorylated Akt. The chloroquine-treated Bcap-37 cells exhibited a marked decrease in the level of mitochondrial transmembrane potential (DeltaPsim), which was accompanied by the activation of caspase-3 and cleaved poly(ADP-ribose) polymerase (PARP). Exposure of Bcap-37 cells to chloroquine also resulted in the induction of spindle abnormalities. In conclusion, the findings in this study suggested that chloroquine might have potential anticancer efficacy, which could be attributed, in part, to its proliferation inhibition and apoptosis induction of cancer cells through modulation of apoptosis and cell cycle-related proteins expressions, down-regulation of mitochondrial transmembrane potential (DeltaPsim), and induction of spindle abnormalities.

[Application of contralateral supplementary puncture in unilateral percutaneous vertebroplasty for poor bone cement dispersion].

OBJECTIVE: To evaluate the clinical value of contralateral supplementary puncture in unilateral percutaneous vertebroplasty(PVP) with poor cement dispersion. METHODS: From January 2015 to December 2016, PVP was performed unilaterally in 285 patients(319 vertebrae) with osteoporotic compression fractures(OVCF). Contralateral percutaneous puncture was performed in 13 patients with poor cement dispersion. Among the patients, 5 cases were male and 8 patients were female, ranging in age from 63 to 88 years old; 1 case of T11, 4 cases of T12, 3 cases of L1, 2 cases of L2, 1 case of L3, and 2 cases of L5. The time from injury to operation ranged from 1 to 16 days. The VAS score, ODI score, residual low back pain and loss of vertebral height were observed. RESULTS: The VAS score and ODI score of 13 patients who underwent contralateral supplementary puncture were significantly improved (P<0.01), and there was no postoperative residual low back pain. During the follow-up period, there was no significant difference in the height loss between the two sides of the vertebral body (0.35 to 3.69 mm on the original puncture side and 0.59 to 3.66 mm on the supplementary puncture side). CONCLUSIONS: For unilateral PVP with poor cement dispersion, contralateral puncture can reduce the occurrence of postoperative residual pain; at the same time, can reduce the difference between the loss of height on both sides of the vertebral body, and then prevent the occurrence of lateral vertebral deformity, which is a safe and reliable method.

[Progress in the design of selective ATP-competitive kinase inhibitors].

Kinases play crucial roles in the life of cell. Their functional abnormity usually leads to many human major diseases including tumors. The prospecting of ATP-competitive small-molecule kinase inhibitors targeting kinases of therapeutic interest has become the focus of researches. Due to the high conservation of the catalytic domain of kinases, the selectivity of kinase inhibitors is poor in general. However, along with the development of structural biology and computer-aided drug design, great progress in the research of selective, ATP-competitive kinase inhibitors has been achieved in recent years. In this account, the review has been made on the development of the design of selective kinase inhibitors.

Antitumor and antimetastatic activities of chloroquine diphosphate in a murine model of breast cancer.

Metastatic breast cancers are hard to treat and almost always fatal. Chloroquine diphosphate, a derivative of quinine, has long been used as a potent and commonly used medicine against different human diseases. We therefore investigated the effects of chloroquine diphosphate on a highly metastatic mouse mammary carcinoma cell line. In vitro treatment of 4T1 mouse breast cancer cells with chloroquine diphosphate resulted in significant inhibition of cellular proliferation and viability, and induction of apoptosis in 4T1 cells in a time- and dose-dependent manner. Further analysis indicated that induction of apoptosis was associated with the loss of mitochondrial membrane potential, release of cytochrome c, and activation of caspase-9 and caspase-3, and cleavage of poly(ADP-ribose) polymerase. The effect of chloroquine diphosphate was then examined using a mice model in which 4T1 cells were implanted subcutaneously. Chloroquine diphosphate (25mg/kg and 50mg/kg, respectively) significantly inhibited the growth of the implanted 4T1 tumor cells and induced apoptosis in the tumor microenvironment. Moreover, the metastasis of tumor cells to the lungs was inhibited significantly and the survival of the mice enhanced. These data suggested that chloroquine diphosphate might have chemotherapeutic efficacy against breast cancer including inhibition of metastasis.

Pharmacophore modelling and virtual screening for identification of new Aurora-A kinase inhibitors.

Aurora-A has been identified as one of the most attractive targets for cancer therapy and a considerable number of Aurora-A inhibitors have been reported recently. In order to clarify the essential structure-activity relationship for the known Aurora-A inhibitors as well as identify new lead compounds against Aurora-A, 3D pharmacophore models were developed based on the known inhibitors. The best hypothesis, Hypo1, was used to screen molecular structural databases, including Specs and China Natural Products Database for potential lead compounds. The hit compounds were subsequently subjected to filtering by Lipinski's rules and docking study to refine the retrieved hits and as a result to reduce the rate of false positive. Finally, 39 compounds were purchased for further in vitro assay against several human tumour cell lines including A549, MCF-7, HepG2 and PC-3, in which Aurora-A is overexpressed. Two compounds show very low micromolar inhibition potency against some of these tumour cells. And they have been selected for further investigation.