Glioblastoma is the most aggressive cerebral gliomas. Despite advances in therapies, the prognosis is still very poor. Therefore, novel therapeutic strategies are required. As a proteasome inhibitor, bortezomib has shown its efficacy as an active antitumor agent against a variety of tumors. However, inhibition of proteasome activity leads to cell death and also induces cell autophagy, and due to the dual roles of autophagy in the survival and death of tumor cells, the effect of inhibition of autophagy on glioblastoma cells remains to be explored. We therefore assessed whether bortezomib is capable of inducing autophagy, and investigated the antitumor effect of bortezomib combined with autophagy inhibitors on human glioblastoma U251 and U87 cells. Cell viability was measured by MTT assay. The expressions of autophagy and apoptosis-related proteins were determined by Western blot analysis. U251 and U87 cells proliferation was inhibited in a dose-dependent manner. Both apoptosis and autophagy induced by bortezomib were observed in human glioblastoma U87 and U251 cells. However, when U251 and U87 cells were co-treated with bortezomib and autophagy inhibitors 3-MA or Atg7 siRNA, the autophagy inhibitors blocked the autophagy in the cells and resulted in a further inhibition of cell proliferation and a further increase in cell apoptosis as compared with that treated with bortezomib alone. These findings indicated that combination of bortezomib and autophagy inhibitors may shed new light on glioblastoma treatment.

Eucommia leaves are dry leaves of Eucommia ulmoides which have long been considered as a functional health food for the treatment of hypertension, hypercholesterolemia, fatty liver, and osteoporosis. With the recent development of Chinese medicine, Eucommia leaves are widely used for tonifying the kidneys and strengthening bone. However, the specific molecular mechanism of Eucommia leaves for strengthening bone remains largely unknown. Osteoblasts are the main functional cells of bone formation; thus, it is essential to study the effect of Eucommia leaves on osteoblasts to better understand their mechanism of action. In the present study, we prepared an aqueous extract of Eucommia leaves (ELAE) and determined its content by high-performance liquid chromatography (HPLC). The effects of ELAE on MC3T3-E1 cells were investigated by CCK-8 assay, alkaline phosphatase (ALP), and Alizarin red S staining assays, combined with RNA sequencing (RNA-seq) and qRT-PCR validation. We demonstrated that ELAE had a significant promoting effect on the proliferation of MC3T3-E1 cells and significantly enhanced extracellular matrix synthesis and mineralization, which were achieved by regulating various functional genes and related signaling pathways. ELAE significantly increased the expression level of genes promoting cell proliferation, such as Rpl10a, Adnp, Pex1, Inpp4a, Frat2, and Pcdhga1, and reduced the expression level of genes inhibiting cell proliferation, such as Npm1, Eif3e, Cbx3, Psmc6, Fgf7, Fxr1, Ddx3x, Mbnl1, and Cdc27. In addition, ELAE increased the expression level of gene markers in osteoblasts, such as Col5a2, Ubap2l, Dkk3, Foxm1, Col16a1, Col12a1, Usp7, Col4a6, Runx2, Sox4, and Bmp4. Taken together, our results suggest that ELAE could promote osteoblast proliferation, differentiation, and mineralization and prevent osteoblast apoptosis. These findings not only increase our understanding of ELAE on the regulation of bone development but also provide a possible strategy to further study the prevention and treatment of osteogenic related diseases by ELAE.

Oxidative stress is increasingly recognized as a risk factor associated with the development and progression of osteoporosis. Fufang Lurong Jiangu Capsule (FLJC) has a known anti-osteoporotic effect, but its pharmacological effect on osteoblasts is not clearly understood. This study was designed to investigate FLJC effects/mechanisms on in vitro hydrogen peroxide (H₂O₂)-induced oxidative damage of osteoblasts and on in vivo lipopolysaccharide (LPS)-induced mice bone loss. FLJC alleviates osteoporosis via unknown pharmacological mechanisms. Chemical compositions of FLJC preparations were analyzed using high-performance liquid chromatographic fingerprinting. After rat bone marrow mesenchymal stem cell differentiation induction, resulting osteoblasts received various 48 h FLJC pretreatments before H₂O₂-based (200 μM) oxidative stress exposure. FLJC effects were measured on osteoblast cell viability, morphological changes, levels of intracellular reactive oxygen species (ROS), localization of mitochondria, activity of antioxidant enzymes, alkaline phosphatase (ALP) and mineralization, the secretion of Col I and expression of osteogenic markers. The percentages of apoptosis were determined by flow cytometric analysis; apoptosis-related protein levels, including nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and heme oxygenase-1 (HO-1) with or without Nrf2 inhibitor were analyzed via western blot. Hematoxylin and eosin (H&E) and ALP staining revealed in vivo FLJC effect on mice LPS-induced bone loss. Five chemical components in FLJC were identified, and fingerprint analysis showed good reproducibility. FLJC pretreatment significantly reduced H₂O₂-induced ROS levels in osteoblasts and increased antioxidant enzyme activities to reduce oxidative damage. With regard to osteoblast differentiation, FLJC pretreatment increased ALP expression, as well as levels of mineralization and osteoblast markers. Additionally, FLJC protected against H₂O₂-induced apoptosis by inhibiting changes in expression of major Bcl-2 family effector proteins of the mitochondrial apoptosis pathway. Furthermore, FLJC protected cells from H₂O₂-induced oxidative damage by up-regulating Nrf2 and HO-1 protein levels. Finally, we confirmed that FLJC administration could reverse the bone loss in LPS-induced mice. These results indicate that FLJC may significantly attenuate oxidative damage of osteoblasts induced by H₂O₂ via the Nrf2/HO-1 signaling pathway, providing new insights to guide development of treatments for osteoporosis induced by oxidative injury.

Prospective radiographic study. To investigate the feasibility of controlling quality of reconstructed sagittal balance for sagittal imbalance. Patients with sagittal imbalance cannot walk or stand erect without overwork of musculature because of compromised biomechanical advantage. The result is muscle fatigue and activity-related pain. During reconstructive surgery, restoration of optimal sagittal balance is crucial for obtaining satisfactory clinical results. However, there is no way to control quality of reconstructed sagittal balance before or during surgery. A method was developed to determine the lumbosacral curve in a way that theoretically would bring sagittal balance to an ideal state by calculation and simulation for each patient before surgery and then template rods of the curve and a blueprint were made accordingly for operative procedures. Ninety-four consecutive patients with sagittal imbalance due to lumbar kyphosis were treated for intractable pain and then followed up for a mean of 4.3 years. Radiographs were analyzed before surgery, 2 months after surgery, and at most recent follow-up. The mean estimated values of L1-S1 lordosis, sacral inclination angle, sacrofemoral distance, and distribution of L1-S1 lordosis at the closing-opening wedge osteotomy site and L4-S1 segments were 30.8°, 24.6°, 0 mm, 16.1% (-5°), and 62% (-19°), respectively. The mean reconstructed values were 41.1°, 23.3°, 3.9 mm, 41% (-17°), and 46% (-19°), respectively. There were significant differences between estimated and reconstructed values of L1-S1 lordosis and the percentage of distributions; however, there was no significant difference between the estimated and reconstructed magnitude of L4-S1 lordosis, sacral inclination angle, and sacrofemoral distance. A properly oriented pelvis can be brought nearly directly above the hip axis. The mean sagittal global balance, represented by the distance between the vertical line through the hip axis and sacral promontory, improved from 61.4 mm before surgery to 3.9 mm 2 months after surgery, and 1.3 mm at final follow-up. Normal sagittal global balance was reconstructed and maintained. The mean sagittal spinal balance measured as the horizontal distance between the C7 sagittal plumb line and the posterior superior corner of S1 improved from 97.4 mm before surgery to 11 mm 2 months after surgery. However, there was significant loss of sagittal spinal balance to 25.4 mm at the fi nal visit. Normal sagittal spinal balance was reconstructed and appeared to be maintained. The magnitude of T1-T12 kyphosis compensated from 13° before surgery to 25.2° 2 months after surgery and 34.5° at fi nal follow-up. Quality control of the reconstructed sagittal balance for sagittal imbalance is possible. Correctly orienting the pelvis, reconstructed by the restoration of enough L1-S1 lordosis with adequate distribution at L4-S1 segments, is a matter of critical importance for optimizing reconstructed sagittal balance. The correctly oriented pelvis can be determined before surgery. Preventing junctional fracture and persistent rehabilitation of surgically injured lumbar extensor musculature are crucial for maintaining the reconstructed sagittal balance.

Study Design. Retrospective radiographic review. Objective. To evaluate the outcome of selective thoracic fusion (STF) by using cantilever bending technique (CBT) and the direct vertebral rotation (DVR) technique for major thoracic-compensatory lumbar (MTCL) curves selected by new curve criteria, which are broader than Lenke curve criteria for STF. Summary of Background Data. Surgical treatment of MTCL curves aims to maximize the number of MTCL curves that can be treated with STF and optimize instrumented thoracic and spontaneous lumbar correction. Comparing current guidelines for STF shows that the surgical technique utilized for STF may affect the curve criteria for MTCL curves for successful STF and thoracic and lumbar correction. Methods. Seventy-eight consecutive idiopathic scoliosis patients with major thoracic-compensatory “C” modifier lumbar curves who met the following three criteria: (1) main thoracic curve (MT) to compensatory lumbar curve (CL) ratios of Cobb magnitude and apical vertebral translation (AVT) greater than one; (2) MT/CL ratio of flexibility less than one; (3) Cobb magnitude of lumbar curve less than 35° on side bending, were treated with STF by using CBT and DVR. Radiographs were analyzed before surgery, immediately after surgery, and at the most recent follow-up (range, 2–5 years). Results. All 78 MTCL curves were successfully treated with STF by using CBT and DVR. A mean 61% thoracic correction was matched by 55% lumbar correction at the most recent follow-up. Spontaneous correction of lumbar AVT occurred in all patients. Global coronal imbalance was common before surgery (mean, 14 mm) and remained so after surgery (mean, 12 mm). There were 49 MTCL curves that did not meet Lenke curve criteria for STF. All were successfully treated with STF by using CBT and DVR. Among these 49 MTCL curves, there were 14 Lenke 1C and 18 Lenke 2C curves with one or two, or all of MT/CL ratios of Cobb magnitude, AVT, and apical vertebral rotation of 1.2 or less, and 6 Lenke 3C and 11 Lenke 4C curves with the Cobb magnitude of residual lumbar curve on side bending between 25° and 35°. Conclusion. CBT and DVR can broaden the current curve criteria of MTCL curves for STF to have more MTCL curves treatable with STF and optimize instrumented thoracic and spontaneous lumbar correction. A more effective surgical technique can not only improve instrumented thoracic and spontaneous lumbar correction but also can broaden the MTCL curve criteria for STF to have more MTCL curves treatable with STF.