[Guilu Erxian Glue Induced Osteogenic Differentiation of BMMSCs and Its Effects on Wnt Signal Pathway].

Objective To observe the effects of Guilu Erxian Glue (GEG) containing serum on osteogenin differentiation of bone marrow mesenchymal stem cells (BMMSCs) and Wnt signal pathway related factors. Methods Totally 100 three months old female SD rats had their bilateral ovaries excised peritoneally They were divided into the low, middle, high GEG groups, and the blank control group by random digit table, 25 in each group. The dose of GEG was calculated according to body surface area, and GEG containing serum was administered by gastrogavage for 7 successive days. Blood was collect- ed by abdominal aorta to prepare drug containing serum. F3 passage BMMSCs of 1-month SD rats were i- solated by whole bone marrow adherent method, and cultured in vitro for 3 passages. The cell surface markers (CD45 and CD90) of F3 passage were detected by flow cytometry (FCM). BMMSCs were trea- ted with different concentrations GEG containing serum for 72 h. Then the cell cycle was determined by FCM, and the proliferation index calculated. The optimal intervention concentration was determined. Then F3 passage BMMSCs were divided into four groups, i.e., the fetal bovine serum (FBS) group, the blank control group, the GEG group, the classical induction group. After they were treated with corresponding medium for 21 days, BMMSCs were dyed with alizarin red staining (ARS) to observe their osteogenin dif- ferentiation. mRNA expressions of alkaline phosphatase (ALP) and osteocalcin (OC) of BMMSCs were detected by RT-PCR. mRNA and protein expressions of Wnt5a, ß-catenin, and lymphoid enhancer factor- 1 (Lef-1) were detected by RT-PCR and Western blot. Results The ratio of CD45 positive expression was 1. 46% ?0. 23%, and the ratio of CD90 positive expression was 96. 97% ±3. 21%. Middle EGE contai- ning serum (10%) could significantly stimulate the proliferation of BMMSCs. In ARS citrus red calcium nodules could be seen in the GEG group and the classical induction group. Compared with the FBS group and the blank control group, mRNA expressions of OC and ALP were up-regulated, mRNA and protein expressions of Wnt5a and p-catenin were up-regulated in the GEG group and the classical induction group (P<0. 05). Compared with the FBS group, the blank control group, and the classical induction group, mRNA and protein expressions of Lef-1 were up-regulated in the EGE group (P <0. 05). Compared with the FBS group and the blank control group, protein expressions of Lef-1 increased in the classical induc- tion group (P <0. 05). Conclusions GEG containing serum had the functions of stimulating the prolifera- tion of BMMSCs, and inducing the osteogenic differentiation of BMMSCs. Its mechanism might be possi- bly related with regulating Wnt signal pathway related factors.

Wireless Optogenetic Targeting Nociceptors Helps Host Cells Win the Competitive Colonization in Implant-Associated Infections.

The role of nociceptive nerves in modulating immune responses to harmful stimuli via pain or itch induction remains controversial. Compared to conventional surgery, various implant surgeries are more prone to infections even with low bacterial loads. In this study, an optogenetic technique is introduced for selectively activating peripheral nociceptive nerves using a fully implantable, wirelessly rechargeable optogenetic device. By targeting nociceptors in the limbs of awake, freely moving mice, it is found that activation induces anticipatory immunity in the innervated territory and enhances the adhesion of various host cells to the implant surface. This effect mediates acute immune cell-mediated killing of Staphylococcus aureus on implants and enables the host to win "implant surface competition" against Staphylococcus aureus. This finding provides new strategies for preventing and treating implant-associated infections.

No genetic causal association between Alzheimer's disease and osteoporosis: A bidirectional two-sample Mendelian randomization study.

Objective: Many observational studies have found an association between Alzheimer's disease (AD) and osteoporosis. However, it is unclear whether there is causal genetic between osteoporosis and AD. Methods: A two-sample Mendelian randomization (MR) study was used to investigate whether there is a causal relationship between osteoporosis and AD. Genes for osteoporosis and AD were obtained from published the genome-wide association studies (GWAS). Single nucleotide polymorphisms (SNPs) with significant genome-wide differences (p < 5 × 10-8) and independent (r 2 < 0.001) were selected, and SNPs with F ≥ 10 were further analyzed. Inverse variance weighted (IVW) was used to assess causality, and the results were reported as odds ratios (ORs). Subsequently, heterogeneity was tested using Cochran's Q test, pleiotropy was tested using the MR-Egger intercept, and leave-one-out sensitivity analysis was performed to assess the robustness of the results. Results: Using the IVW method, MR Egger method, and median-weighted method, we found that the results showed no significant causal effect of osteoporosis at different sites and at different ages on AD, regardless of the removal of potentially pleiotropic SNPs. The results were similar for the opposite direction of causality. These results were confirmed to be reliable and stable by sensitivity analysis. Conclusion: This study found that there is no bidirectional causal relationship between osteoporosis and AD. However, they share similar pathogenesis and pathways.

[Prevention of dura adherence in spinal canal after microendoscopic discectomy by different methods: a clinical study of 165 cases].

OBJECTIVE: To evaluate the effects of different methods in prevention of post-operational scar formation and dura adherence in the spinal canal after microendoscopic discectomy (MED). METHODS: 165 patients undergoing MED were randomly divided into 3 equal groups: Group A, with the yellow ligament dissected and with the space between vertebral laminae sprinkled with sodium hyaluronate before the closing of the incision; Group B, with the yellow ligament reserved; and Group C, with the yellow ligament reserved and with the space between vertebral laminae sprinkled with sodium hyaluronate before the closing of the incision. All the patients were followed up 2, 4, and 8 weeks, and 1 and 2 years after the operation. Japanese Orthopedic Association (JOA) scoring system was used to evaluate the outcomes. RESULTS: The JOA scores 2 weeks after MED were not significantly different among the 3 groups; and from then on the JOA scores of Groups B and C were all higher than those of group A (A and B: t(4w) = 0.602, t(8w) = 0.701, t(1y) = 0.623, t(2y) = 0.654; A and C: t(4w) = 0.833, t(8w) = 0.759, t(1y) = 0.714, t(2y) = 0.771, all P < 0.05), however, there were not significantly differences at all time points between Groups B and C (B and C: t(2w) = 0.041, t(4w) = 0.135, t(8w) = 0.980, t(1y) = 0.530, t(2y) = 0.103, all P > 0.05). CT showed that a great amount of scar was seen, surrounding the dura mater sac and nerve roots in Group A, and there was a great amount of scar outside the yellow ligament and no remarkable compression of dura mater sac and nerve roots in Groups B and C. CONCLUSION: Reservation of yellow ligament effectively prevents scar adhesion inside the vertebral canal after MED. Sprinkling of sodium hyaluronate is also effective, however, its effect only lasts a short time.

Correlation of bone fragments reposition and related parameters in thoracolumbar burst fractures patients.

The aim of this study is to determine if thoracolumbar vertebral body collapse or canal compromise (CC) is associated with reposition of bone fragment. We retrospective review medical charts of patients with thoracolumbar burst fractures from July 2010 to September 2013. The fractures were classified according to the Arbeit Fuer Osteoosynthese (AO) classification system. Neurological status was classified according to American Spinal Injury Association (ASIA). Patients were divided into two groups (reposition group and non-reposition group) according to whether the bone fragments were reposition or non-reposition after surgery. Mimics measured mid-sagittal canal diameter (MSD), transverse canal diameter (TCD), local kyphosis (LK) and calculated anterior vertebral body compression ratio (AVBCR), middle vertebral body compression ratio (MVBCR), posterior vertebral body compression ratio (PVBCR), and mid-sagittal canal diameter compression ratio (MSDCR) on the preoperative CT image. The results indicated that 55 patients were included in the study. There are 35 patients with reposition of bone fragment and 20 patients with non-reposition of bone fragment after surgery. There were significant difference on MSD (t = 3.258, P = 0.002), TCD (t = 2.197, P = 0.032), AVBCR (t = -2.063, P = 0.044), MVBCR (t = -2.526, P = 0.015), PVBCR (t = -2.211, P = 0.031), MSDCR (t = -4.975, P = 0.000) between two groups before surgery. There was a significant correlation between reposition of bone fragment and AO classification (OR = 5.251, P = 0.022), and MSDCR (OR = 7.366, P = 0.007). There was no significant correlation between reposition and AVBCR, MVBCR, PVBCR, LK, MSD and TCD. In conclusion, this study indicates that AO classification and MSDCR are predictors of reposition of bone fragment.

Pien Tze Huang induces apoptosis in multidrug­resistant U2OS/ADM cells via downregulation of Bcl­2, survivin and P-gp and upregulation of Bax.

Pien Tze Huang (PZH) is a well-known traditional Chinese formula that was first prescribed by a royal physician in the Ming Dynasty. PZH has been used to treat various types of cancers including osteosarcoma. Previous studies have shown that PZH may effectively inhibit osteosarcoma cell growth in vivo and in vitro via induction of apoptosis and inhibition of migratory and invasive abilities. However, little is known regarding the effects of PZH on osteosarcomas that are resistant to chemotherapy, which has emerged as a major clinical problem. In the present study, the cellular effects of PZH on multidrug-resistant U2OS/ADM human osteosarcoma cells were investigated. Our results showed that PZH reduced cell viability in a dose- and time-dependent manner and arrested cells in the G2/M phase of the cell cycle, suggesting that PZH inhibits the proliferation of U2OS/ADM cells. Hoechst 33258 staining and Annexin V/propidium iodide double staining revealed typical nuclear features of apoptosis, and treatment with PZH increased the proportion of apoptotic Annexin V-positive cells in a dose-dependent manner. Further experiments demonstrated that apoptosis induction by PZH was accompanied by downregulation of Bcl-2 and survivin and upregulation of Bax. In addition, following treatment with PZH, intracellular Rhodamine 123 accumulation was increased and the expression of P-gp was significantly suppressed. Taken together, these results provide a possible molecular mechanism for the anticancer effect of PZH on U2OS/ADM cells and suggest that PZH may be a potent therapeutic agent for drug-resistant osteosarcoma.