miR-150 is downregulated in osteosarcoma and suppresses cell proliferation, migration and invasion by targeting ROCK1.

[This retracts the article DOI: 10.3892/ol.2017.5709.].

Proton density fat fraction measurements of rotator cuff muscles: Accuracy, repeatability, and reproducibility across readers and scanners.

PURPOSE: To determine the accuracy, repeatability, and reproducibility of magnetic resonance imaging-based proton density fat fraction (MRI-PDFF) measurements of rotator cuff muscles between two readers and three different scanners. METHODS: Twenty-seven volunteers underwent serial shoulder MRI examinations of both left and right sides on one 1.5-T MRI scanner and two 3.0-T MRI scanners. Two independent readers measured muscular PDFF of the supraspinatus, infraspinatus/teres minor muscle, and subscapularis. MR spectroscopy-based proton density fat fraction (MRS-PDFF) was regarded as the reference standard for assessing accuracy. A "coffee break" examination method was used to test the repeatability of each scanner. Bland-Altman plots, Pearson correlation, and linear regression analysis were used to assess bias and linearity. The Wilcoxon signed-rank test and Friedman test were applied to evaluate repeatability and reproducibility. RESULTS: MRI-PDFF measurements indicated strong linearity (R2 = 0.749) and small bias (-0.18%) in comparison with the MRS-PDFF measurements. A very strong positive Pearson correlation (r = 0.955-0.986) between the PDFF estimates of the two repeat scans indicated excellent repeatability. The PDFF measurements showed high reproducibility, with a strong positive Pearson correlation (r = 0.668-0.698) and a small mean bias (-0.04 to -0.10%) across different scanners. CONCLUSION: MRI-PDFF measurements of rotator cuff muscles were highly accurate, repeatable, and reproducible across different readers and scanners, leading us to the conclusion that PDFF can be a reliable and robust quantitative imaging biomarker for longitudinal or multi-center studies.

Ghrelin infusion into the basolateral amygdala suppresses CTA memory formation in rats via the PI3K/Akt/mTOR and PLC/PKC signaling pathways.

Ghrelin is a circulating orexigenic hormone that promotes feeding behavior and regulates metabolism in humans and rodents. We previously reported that local infusion of ghrelin into the basolateral amygdala (BLA) blocked memory acquisition for conditioned taste aversion (CTA) by activating growth hormone secretagogue receptor 1a. In this study, we further explored the underlying mechanism and signaling pathways mediating ghrelin modulation of CTA memory in rats. Pharmacological agents targeting distinct signaling pathways were infused into the BLA during conditioning. We showed that preadministration of the PI3K inhibitor LY294002 abolished the repressive effect of ghrelin on CTA memory. Moreover, LY294002 pretreatment prevented ghrelin from inhibiting Arc and zif268 mRNA expression in the BLA triggered by CTA memory retrieval. Preadministration of rapamycin eliminated the repressive effect of ghrelin, while Gsk3 inhibitors failed to mimic ghrelin's effect. In addition, PLC and PKC inhibitors microinfused in the BLA blocked ghrelin's repression of CTA acquisition. These results demonstrate that ghrelin signaling in the BLA shapes CTA memory via the PI3K/Akt/mTOR and PLC/PKC pathways. We conducted in vivo multichannel recordings from mouse BLA neurons and found that microinjection of ghrelin (20 µM) suppressed intrinsic excitability. By means of whole-cell recordings from rat brain slices, we showed that bath application of ghrelin (200 nM) had no effect on basal synaptic transmission or synaptic plasticity of BLA pyramidal neurons. Together, this study reveals the mechanism underlying ghrelin-induced interference with CTA memory acquisition in rats, i.e., suppression of intrinsic excitability of BLA principal neurons via the PI3K/Akt/mTOR and PLC/PKC pathways.

Nanocarrier-based activation of necroptotic cell death potentiates cancer immunotherapy.

Even though immunological checkpoint inhibitors have demonstrated a potent anti-tumor effect in clinical practice, the low immunogenicity of the majority of tumors still results in a lower response rate and a higher resistance to mono-immunotherapy. Recent studies revealed that immunogenic cell death (ICD) augments T cell responses against some cancers, thus indicating that this combination therapy may further improve the anti-tumor immunity produced by anti-PD-1/PD-L1. Herein a robust synergetic strategy is reported to integrate the activation of necroptotic cell death and the subsequent using of immune checkpoint inhibitors. Liposomes have good biocompatibility and are widely used as drug carriers. Using liposomes as TNF-α-loaded nanoplatforms achieves in vivo tumor targeting and long-term retention in the tumor microenvironment. Tumor cells treated with TNF-α-loaded liposomes exhibited the hallmarks of ICD including the release of high mobility group box 1 (HMGB1) and lactate dehydrogenase (LDH). Additionally, the tumor cell necrosis caused by TNF-α induces the in situ release of tumor-specific antigens, thus increasing the dendritic cell (DC) activation and T cell infiltration when combined with the checkpoint blockade therapy. Collectively, significant tumor reduction is accomplishable by this synergetic strategy, in which TNF-α-loaded liposomes convert the tumor cell into an endogenous vaccine and improve the anti-tumor immunity of anti-PD-1/PD-L1.

Development and validation of a nomogram to predict perioperative blood transfusion in patients undergoing total knee arthroplasty.

BACKGROUND: The need for a transfusion is one of the adverse events following total knee arthroplasty (TKA), and accurately predicting this need remains challenging for arthroplasty surgeons. The purpose of the present research is to study the preoperative predictors of transfusion risk in patients following TKA and develop a nomogram. METHODS: The nomogram was developed based on a training set of 5402 patients who underwent TKA at the Affiliated Hospital of Qingdao University between September 2013 and November 2018. The independent predictors of transfusion were identified by univariate, LASSO, and binary logistic regression analyses. Then, a nomogram was established based on these independent predictors. The area under the curve (AUC), calibration curve, and decision curve analysis (DCA) were selected to evaluate the nomogram. The results were validated using an independent set of 1116 patients who underwent TKA between December 2018 and September 2019. In addition, we also carried out subgroup analyses in the training and testing sets based on the independent predictors. RESULTS: Five independent predictors were identified by multivariate analysis and were used to establish the nomogram. The AUCs of the nomogram were 0.884 (95% CI: 0.865-0.903) and 0.839 (95% CI, 0.773-0.905) in the training and testing sets, respectively. In both the training and testing sets, the calibration curve indicated that the prediction by the nomogram was highly consistent with the actual observation, and the DCA indicated that the nomogram had a favorable level of clinical usefulness. In addition, the AUC of the nomogram was significantly higher than the AUC of any independent predictor for predicting transfusion risk following TKA, and the subgroup analysis showed good performance in 20 subgroups. CONCLUSION: Lower preoperative Hb levels, simultaneous bilateral TKA, lower BMI, older age, and coronary heart disease were identified as independent predictors of postoperative transfusion in patients following TKA. A nomogram incorporating the above five predictors could accurately predict the transfusion risk.

Proximal fibular osteotomy alleviates medial compartment knee osteoarthritis in a mouse model.

PURPOSE: The purpose of this study was to establish a mouse model of proximal fibular osteotomy (PFO), and to determine if PFO could delay degeneration of the medial compartment of the knee joint in a mouse model. METHODS: An animal model of destabilization of the medial meniscus (DMM) was used to induce post-traumatic knee osteoarthritis (OA). PFO was performed to examine the effectiveness of PFO on protection against medial compartment knee OA. Micro-CT was used to observe osteosclerosis development in the subchondral bone, and Safranin O-fast green staining was used to evaluate the progression of articular cartilage destruction. The condylar-plateau angle (CPA) and anatomical femorotibial angle (aFTA) were measured to determine whether knee alignment was changed after PFO. RESULTS: PFO treatment could decrease osteophyte formation and osteosclerosis development in the subchondral bone, as observed by micro-CT. The value of the ratio of trabecular bone volume to total volume (BV/TV) of DMM+PFO group was lower than that of DMM group. PFO also inhibited the progression of articular cartilage destruction. DMM + PFO group displayed decreased maximal and summed OA scores, as compared with DMM group. Moreover, the change of knee alignment was reduced by PFO, which might be the mechanism of PFO alleviating medial compartment knee OA. CONCLUSION: Our results indicated that PFO could alleviate medial compartment knee OA in a mouse model.

miR-150 is downregulated in osteosarcoma and suppresses cell proliferation, migration and invasion by targeting ROCK1.

Osteosarcoma (OS) is the most common form of bone malignancy in children and adolescents. A class of molecules known as microRNAs (miRNAs) have been routinely associated in the development and progression of OS. The present study was centered on the less well-known miRNA, miRNA (miR)-150, and its role in OS was investigated. The levels of miR-150 were examined in 40 tissue specimens from patients with OS and adjacent normal tissues using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis. In addition the expression levels of miR-150 were examined in three OS cell lines and a normal osteoblast cell line. Cell proliferation, migration and invasion assays were performed to establish the correlation between miR-150 and metastasis. The potential targets of miR-150 were theoretically predicted and one high-scoring target, Rho-associated kinase 1 (ROCK1), was established to be a direct target using RT-qPCR and western blot analyses and Pearson's correlation analysis. The results indicated that miR-150 was downregulated in tissues from patients with OS and cell lines. Secondly, it was shown that the overexpression of miR-150 was inversely correlated with OS cell proliferation, migration and invasion. It was also shown that miR-150 negatively regulated the gene expression of ROCK1 in the OS cell lines. Finally, the interaction between miR-150 and ROCK1 was established and it was shown that miR-150 directly targeted ROCK1. In conclusion, miR-150 was found to be a tumor suppressor, and the suppression of miR-150 resulted in elevation in the levels of ROCK1. This interaction between miR-150 and ROCK1 may be key in the progression of OS. Furthermore, miR-150 or ROCK1 may be potential therapeutic targets for the treatment of OS.

Immune therapy with cultured microglia grafting into the injured spinal cord promoting the recovery of rat's hind limb motor function.

OBJECTIVE: To study the effect of activated microglia grafting on rats' hind limb motor function recovery after spinal cord injury. METHODS: Microglia were separated from primary culture and subcultured for 3 generations. Lipopolysaccharide was added to the culture medium with the terminal concentration of 10 microl/L for microglia activation 3 days before transplantation. Totally 80 adult Wistar rats were divided into transplantation group and control group, with 40 rats in each group. Spinal cord injury model of rats was set by hitting onto the spinal cord using a modified Allen impactor. With a 5 microl micro-syringe, the activated microglia suspension was injected into the injured area 7 days after the first operation. Basso, Beattie and Bresnahan (BBB) scoring for hind limb motor function was taken on the 1st, 7th, 14th, 21st, and 28th day after microglia transplantation, and 8 rats were sacrificed at each time point mentioned above, respectively. Frozen sections of the spinal cord were made for haematoxylin-eosin (HE) and Naoumenko-Feigin stainings. SPSS 11.0 software was used for statistical analysis. RESULTS: BBB scores for hind limb motor function on the 14th, 21st, and 28th day were significantly higher compared with the control group. Most liquefaction necrosis areas disappeared and only a few multicystic cavities surrounded by aggregated microglia remained in the transplantation group. Naoumenko-Feigin staining for microglia showed that the transplantation group had significantly more positive cells (P < 0.05). CONCLUSIONS: Grafting of activated microglia into the injured spinal cord can significantly promote the hind limb motor function recovery in rats with spinal cord injury and reduce the size of liquefaction necrosis area. The extent of lower limb motor function improvement has a positive correlation with the number of aggregated microglia.

Anterior cruciate ligament reconstruction with bone-patellar tendon-bone autograft versus allograft.

PURPOSE: The purpose of this study was to analyze the clinical outcome of arthroscopic anterior cruciate ligament (ACL) reconstruction with bone-patellar tendon-bone autograft versus allograft. METHODS: Between May 2000 and June 2004, 172 patients undergoing arthroscopic bone-patellar tendon-bone ACL reconstruction were prospectively randomized into autograft (n = 86) or allograft (n = 86) groups. The senior surgeon performed all operations using the same surgical technique. Each fixation was performed by means of an interference screw. Patients were evaluated preoperatively and postoperatively at follow-up. Of the patients, 156 (76 in the autograft group and 80 in the allograft group) were available for full evaluation. Evaluations included a detailed history, physical examination, functional knee ligament testing, KT-2000 arthrometer testing (MEDmetric, San Diego, CA), Harner's vertical jump and Daniel's 1-leg hop tests, Lysholm score, Tegner score, International Knee Documentation Committee standard evaluation form, Cincinnati knee score, and radiograph. RESULTS: Demographic data were comparable between groups. The mean follow-up was 5.6 years for both groups. There were no statistically significant differences according to evaluations of outcome between the 2 groups except that patients in the allograft group had a shorter operation time and longer fever time postoperatively compared with the autograft group. The postoperative infection rates were 0% and 1.25% for the autograft group and allograft group, respectively. There was a significant difference (P < .05) in the development of osteoarthritis between the operated knee in comparison to the contralateral knee according to radiographs. However, no significant difference was found between the 2 groups at the final follow-up examination (P > .05). CONCLUSIONS: Both groups of patients achieved almost the same satisfactory outcomes after a mean of 5.6 years of follow-up. Allograft is a reasonable alternative to autograft for ACL reconstruction. LEVEL OF EVIDENCE: Level II, prospective comparative study.

ACL reconstruction with BPTB autograft and irradiated fresh frozen allograft.

OBJECTIVE: To analyze the clinical outcomes of arthroscopic anterior cruciate ligament (ACL) reconstruction with irradiated bone-patellar tendon-bone (BPTB) allograft compared with non-irradiated allograft and autograft. METHODS: All BPTB allografts were obtained from a single tissue bank and the irradiated allografts were sterilized with 2.5 mrad of irradiation prior to distribution. A total of 68 patients undergoing arthroscopic ACL reconstruction were prospectively randomized consecutively into one of the two groups (autograft and irradiated allograft groups). The same surgical technique was used in all operations done by the same senior surgeon. Before surgery and at the average of 31 months of follow-up (ranging from 24 to 47 months), patients were evaluated by the same observer according to objective and subjective clinical evaluations. RESULTS: Of these patients, 65 (autograft 33, irradiated allograft 32) were available for full evaluation. When the irradiated allograft group was compared to the autograft group at the 31-month follow-up by the Lachman test, the anterior drawer test (ADT), the pivot shift test, and KT-2000 arthrometer test, statistically significant differences were found. Most importantly, 87.8% of patients in the autograft group and just only 31.3% in the irradiated allograft group had a side-to-side difference of less than 3 mm according to KT-2000. The failure rate of the ACL reconstruction with irradiated allograft (34.4%) was higher than that with autograft (6.1%). The anterior and rotational stabilities decreased significantly in the irradiated allograft group. According to the overall International Knee Documentation Committee (IKDC), functional and subjective evaluations, and activity level testing, no statistically significant differences were found between the two groups. Besides, patients in the irradiated allograft group had a shorter operation time and a longer duration of postoperative fever. When the patients had a fever, the laboratory examinations of all patients were almost normal. Blood routine was normal, the values of erythrocyte sedimentation rate (ESR) were 5~16 mm/h and the contents of C reactive protein (CRP) were 3-10 mg/L. CONCLUSION: We conclude that the short term clinical outcomes of the ACL reconstruction with irradiated BPTB allograft were adversely affected. The less than satisfactory results led the senior authors to discontinue the use of irradiated BPTB allograft in ACL surgery and not to advocate using the gamma irradiation as a secondary sterilizing method.

[A study of three different procedures for avulsion fracture of tibial intercondylar eminence].

OBJECTIVE: To observe the clinical outcomes and differences among three surgical procedures for avulsion fracture of tibial intercondylar eminence. METHODS: From October 1995 to October 2005, 3 different procedures had been performed on 49 patients, which included open reduction and internal fixation (Group A, n = 17), arthroscopic reduction and internal fixation( Group B, n = 19) and limited open reduction and internal fixation assisted with arthroscopy (Group C, n = 13). All patients were followed up for 1 to 10 years (average 4. 6 years). RESULTS: For the 3 groups, normal extension function were 35.5%, 16.0% and 38.0%, mild abnormal 35.5%, 11.0% and 23.0%, moderate abnormal 29.0%, 47.0% and 31.0%; and severe abnormal were 0,26.0% and 8.0%. Normal flexion were 82.0%, 78.0% and 84.0%, mild abnormal 12.0%, 11.0% and 8.0%, moderate abnormal 6.0%, 11.0% and 8.0%. The positive rate of Lachman's or anterior drawer test were 35.0%, 45.0% and 38.0%; McIntoshi test were 11.0%, 16.0% and 13.0% respectively for 3 groups. Lysholm' scale were average 98.6, 97.3 and 98.2; Tegner' scale were 6.6, 6.4 and 6.7. KT-2000 showed that anterior translation of tibial were 6.9, 7.1 and 6.6 mm; side to side difference were 11.4, 1.7 and 1.5 mm, except that statistically significant differences were found in extension function between group A and group B (P = 0.02). There were no any statistically significant differences in other aspects. CONCLUSION: Limited open and properly over reduction and three dimensional as well as strong internal fixation assisted with arthroscopy should been attempted for the treatment of avulsion fracture of tibial intercondylar eminence.