CYLD alleviates NLRP3 inflammasome-mediated pyroptosis in osteoporosis by deubiquitinating WNK1.

BACKGROUND: Osteoporosis (OP) is the result of bone mass reduction and bone structure disorder. Bone marrow mesenchymal stem cells (BMSCs) are the main source of osteogenic precursor cells involved in adult bone remodeling. The involvement of the deubiquitinating enzyme CYLD in OP has recently been discovered. However, the detailed role and mechanism of CYLD remain unknown. METHODS: The OP mouse model was established by performing ovariectomy (OVX) on mice. Hematoxylin and eosin staining, Masson and Immunohistochemical staining were used to assess pathologic changes. Real-time quantitative PCR, Western blot, and immunofluorescence were employed to assess the expression levels of CYLD, WNK1, NLRP3 and osteogenesis-related molecules. The binding relationship between CYLD and WNK1 was validated through a co-immunoprecipitation assay. The osteogenic capacity of BMSCs was determined using Alkaline phosphatase (ALP) and alizarin red staining (ARS). Protein ubiquitination was evaluated by a ubiquitination assay. RESULTS: The levels of both CYLD and WNK1 were decreased in bone tissues and BMSCs of OVX mice. Overexpression of CYLD or WNK1 induced osteogenic differentiation in BMSCs. Additionally, NLRP3 inflammation was activated in OVX mice, but its activation was attenuated upon overexpression of CYLD or WNK1. CYLD was observed to reduce the ubiquitination of WNK1, thereby enhancing its protein stability and leading to the inactivation of NLRP3 inflammation. However, the protective effects of CYLD on osteogenic differentiation and NLRP3 inflammation inactivation were diminished upon silencing of WNK1. CONCLUSION: CYLD mitigates NLRP3 inflammasome-triggered pyroptosis in osteoporosis through its deubiquitination of WNK1.

Remodeling Microenvironment for Implant-Associated Osteomyelitis by Dual Metal Peroxide.

Implant-associated osteomyelitis (IAOM) is characterized by bone infection and destruction; current therapy of antibiotic treatment and surgical debridement often results in drug resistance and bone defect. It is challenging to develop an antibiotic-free bactericidal and osteogenic-enhanced strategy for IAOM. Herein, an IAOM-tailored antibacterial and osteoinductive composite of copper (Cu)-strontium (Sr) peroxide nanoparticles (CSp NPs), encapsulated in polyethylene glycol diacrylate (PEGDA) (CSp@PEGDA), is designed. The dual functional CSp NPs display hydrogen peroxide (H2 O2 ) self-supplying and Fenton catalytic Cu2+ ions' release, generating plenty of hydroxyl radical (• OH) in a pH-responsive manner for bacterial killing, while the released Sr2+ promotes the in vitro osteogenicity regarding cell proliferation, alkaline phosphatase activity, extracellular matrix calcification, and osteo-associated genes expression. The integration of Cu2+ and Sr2+ in CSp NPs together with the coated PEGDA hydrogel ensures the stable and sustainable ion release during short- and long-term periods. Benefitted from the injectablity and photo-crosslink ability, CSp@PEGDA is able to thoroughly fill the infectious site and gelate in situ for bacterial elimination and bone regeneration, which is verified through in vivo evaluation using a clinical-simulating IAOM mouse model. These favorable abilities of CSp@PEGDA precisely meet the multiple therapeutic needs and pave a promising way for implant-associated osteomyelitis treatment.

New understandings of the genetic regulatory relationship between non-coding RNAs and m6A modification.

One of the most frequent epigenetic modifications of RNA in eukaryotes is N6 methyladenosine (m6A), which is mostly present in messenger RNAs. Through the influence of several RNA processing stages, m6A modification is a crucial approach for controlling gene expression, especially in cancer progression. It is universally acknowledged that numerous non-coding RNAs (ncRNAs), such as microRNAs, circular RNAs, long non-coding RNAs, and piRNAs, are also significantly affected by m6A modification, and the complex genetic regulatory relationship between m6A and ncRNAs plays a pivotal role in the development of cancer. The connection between m6A modifications and ncRNAs offers an opportunity to explore the oncogene potential regulatory mechanisms and suggests that m6A modifications and ncRNAs could be vital biomarkers for multiple cancers. In this review, we discuss the mechanisms of interaction between m6A methylation and ncRNAs in cancer, and we also summarize diagnostic and prognostic biomarkers for clinical cancer detection. Furthermore, our article includes some methodologies for identifying m6A sites when assessing biomarker potential.

Multiple Reconstructive Osteotomy Treating Malunited Calcaneal Fractures Without Subtalar Joint Fusion.

OBJECTIVE: Various surgical techniques have been reported in treating calcaneal malunions over the decades, while the operations on single plane were accompanied by respective limitations. The purpose of this study is to evaluate the efficacy of a novel multiple reconstructive osteotomy for treating malunited calcaneal fractures without subtalar joint fusion. METHODS: From March 12, 2010, to August 17, 2017, 10 patients (10 feet) with malunited calcaneal fractures were treated with multiple reconstructive osteotomy with subtalar joint-preserving operations. All patients were treated with a corrective osteotomy, joint realignment, soft tissue balancing, and secondary internal fixation at a mean of 5.6 ± 2.41 months since the initial injury. With the utilization of the multiple reconstructive osteotomy, the posterior facet was restored to preserve the subtalar joint. All patients were evaluated clinically and radiographically at a mean follow-up of 3.04 ± 1.21 years. RESULTS: All patients were subjectively satisfied with the treatment. The average time to union was 12.2 ± 1.11 weeks. The American Orthopedic Foot and Ankle (AOFAS) ankle and hind foot score was 86.3 ± 4.45 (t = 27.64, P < 0.0001, paired t-test), which was significantly higher than the preoperative assessment. Postoperative radiographic assessment revealed great improvement in Böhler's angle (from 25.4° to 86.3°), talocalcaneal height (65.15-72.68 mm) and Calcaneus-talus angle (from 34.46° to 39.7°). One patient had mild discomfort after a 1-h brisk walk. One patient was suspected to have early posttraumatic arthritis of the subtalar joint based upon radiographic evidence during the follow-up, but the patients could walk normally for a long time without pain. CONCLUSION: Multiple reconstructive osteotomy is an effective way to restore the calcaneal morphology and preserve the subtalar joint for selected calcaneal malunion.

Malunited calcaneal fracture: the role and technique of osteotomy-a systematic review.

PURPOSE: The research is aimed to introduce various corrective osteotomies utilized in treating calcaneal malunions in published papers, to further analyze the results, and to summarize recommended indications. METHODS: The relevant research screening was conducted on the following search engines: the Cochrane Library, Web of Science, PubMed, Embase, Medline, and Academic Search Premier. Key words input included "calcaneal/calcaneus", "malunion," and "malunited fracture(s)" with Boolean operators "AND" and "OR." The inclusion criteria were researches containing surgical procedures treating calcaneal malunion with corrective osteotomy and published in the English language. For included research article, such information was extracted and analyzed: the type of calcaneal malunion, the time from initial injury to corrective surgery, the method of osteotomy, outcomes of each osteotomy (score systems, Bohler angle, talocalcaneal height and width of calcaneus, etc.), the function of the affected limb, post-operative complications, and patients' satisfaction. For included review, descriptive, commentary, or indicative sentences about corrective osteotomy were highlighted, analyzed, and summarized. RESULTS: Ten research articles (170 patients with 184 feet) and nine reviews were included in this review, presenting seven types of corrective osteotomies (lateral wall exostectomy, Dwyer osteotomy, lateral wedge opening osteotomy, Romash osteotomy, tongue osteotomy, sagittal resection osteotomy, and modified Dwyer osteotomy). CONCLUSION: A different corrective osteotomy with/without arthrodesis is recommended to be utilized based on the classification of the malunion and the condition of the cartilage in treating malunited calcaneal fractures. With adequate postoperative care and rehabilitation, the results of treatment could be associated with patients' satisfaction and good function.

A neglected problem in the utilization of free anterolateral thigh flap toward reconstructing complicated wounds of extremities: the obliteration of deep dead space.

BACKGROUND: Deep dead space may be thought as an independent risk factor of the poor infection control after flap reconstruction in complex limb wounds. But it can be easily neglected. The conventional skin flap and musculocutaneous flap are difficult to obliterate the deep dead space in irregular shape effectively. It was investigated that the clinical application of chimeric anterolateral thigh perforator flap in the treatment of complex wounds complicated with deep dead space of the extremities in the paper. METHODS: Fifty-six cases complicated with deep dead space wounds were registered in group. Following thorough debridement and treatment with VSD, the granulation tissues grew with well-controlled infection. And then the chimeric anterolateral thigh perforator flap was used to obliterate the deep dead space and repair the wounds. The postoperative flap survival and infection conditions were evaluated. RESULTS: Overall, the infection was effectively controlled, without persistent exudation or sinus tract formation after wound healing. While 5 cases lost to follow-up, the remaining 51 cases were followed up until 15 months on average. Generally, the affected extremities recovered satisfactorily with normal appearances and texture of the flaps, along with normal functions. Importantly, no recurrence of infection was observed. CONCLUSION: During the grafting of chimeric perforator flap pedicled with lateral thigh muscle flap, the muscle flap is recommended to obliterate the deep dead space while the skin flap is being used to cover the wound. The combination of these two technologies performed well in the repair and reconstruction of the complex wounds of the extremities, possessing potential for broader clinical application.

Chinese Consensus on Insertional Achilles Tendinopathy.

BACKGROUND: Insertional Achilles tendinopathy (IAT) is a common finding in the clinic. However, consensus on its mechanism, pathological process, diagnosis, treatment, and rehabilitation is lacking. Thus, the Chinese Society of Sports Medicine organized and invited experts representing the fields of ankle disease and tendinopathy to jointly develop an expert consensus on IAT. STUDY DESIGN: A consensus statement of the Chinese Society of Sports Medicine. METHODS: A total of 34 experts in the field of sports medicine and orthopaedics were invited to participate in the compilation of a consensus statement regarding IAT. Consensus was achieved according to the Delphi method. First, 10 working groups composed of 34 experts were established to compile draft statements about clinical problems related to IAT by reviewing and analyzing the available literature. An expert consensus meeting to discuss drafts was then arranged. Each statement was individually presented and discussed, followed by a secret vote. Consensus was reached when more than 50% of the experts voted in its favor. The strength of the proposed recommendation was classified based on the proportion of favorable votes: consensus, 51% to 74%; strong consensus, 75% to 99%; unanimity, 100%. RESULTS: Of the 10 expert consensus statements on the clinical diagnosis and treatment of IAT, there was strong consensus for 8 statements and unanimity for 2 statements. CONCLUSION: This expert consensus focused on the concepts, causes, pathological process, clinical diagnosis, and treatment of IAT. Accepted recommendations in these areas which will assist clinicians in carrying out standardized management of related diseases.

[Progress in treatment of acute achilles tendon rupture].

OBJECTIVE: To review the progress in the treatment of acute Achilles tendon rupture. METHODS: Recent literature about the treatment of acute Achilles tendon rupture was reviewed and analyzed. RESULTS: Treatments of acute Achilles tendon rupture include operative and non-operative treatments. Operative treatments include open surgery and percutaneous minimally invasive surgery. Compared with non-operative treatment, operative treatment can effectively reduce the re-rupture incidence, but it had higher complication incidences of wound infection and nerve injury. Although early functional rehabilitation during non-operative treatment could reduce the re-rutpture incidence, there is no consistent orthopaedic device and guideline for functional rehabilitation. CONCLUSION: Both operative and non-operative treatments have advantages and disadvantages for the treatment of acute Achilles tendon rupture. No consistent conclusion is arrived regarding functional recovery. Future studies should explore the strategy of early functional rehabilitation during non-operative treatment and its mechanism of promoting tendon healing.

Reconstruction of shoulder abduction by multiple nerve fascicle transfer through posterior approach.

PURPOSE: To evaluate the feasibility and clinical efficacy of multiple nerve fascicle transfer through posterior approach for reconstruction of shoulder abduction in patients with C5 or upper brachial plexus injury. METHODS: 11 patients (aged between 17 and 56 years) with dysfunction of shoulder abduction post C5 or upper brachial plexus injury were recruited in this study. Among them, four out of 11 patients also had dysfunction of elbow flexion simultaneously. The duration from injury to the surgery ranged from 4 to 12 months, with an average of 6.7 months. The affected shoulder joints showed abduction, extension and elevation dysfunction, but the muscle strength of shoulder shrugging and elbow extension was graded to M4 or higher. Accessory nerve was transferred to the suprascapular nerve and triceps muscle was branched to the axillary nerve through posterior approach. Ulnar fascicle was transferred to the motor branches of biceps for the 4 patients involved with elbow flexion dysfunction. RESULTS: Ten out of 11 cases were followed-up for 15-36 months. Neo-potential of deltoid and supraspinatus/infraspinatus was documented at 4-5 months post surgery. Shoulder abduction (and elbow flexion) was reanimated at 4-8 months post surgery. Significant improvement was observed at 15-36 months post surgery, shoulder abduction regained to 40-160° (mean: 92.5°), muscle strength of supraspinatus/infraspinatus and deltoid were graded to M3-M5 (mean: 4.0 and 4.1); 3 cases muscle strength of elbow flexion was graded from M4 to M5- (mean: 4.4) with 1 case loss. Shoulder shrugging of trapezius was graded to M5 in 5 cases, M5- in 2 cases, M4 in 2 cases and M3 in 1 case (mean: 4.5). All cases showed normal elbow extension and muscle strength of triceps (M5). CONCLUSION: It is feasible to carry out multiple nerve fascicle transfers for early reconstruction of shoulder abduction by posterior approach. Patients who received this procedure achieved good functional recovery and their donor site morbidity/injury was minimal.

A report of 15 hand allotransplantations in 12 patients and their outcomes in China.

BACKGROUND: Limb allotransplantation is emerging as a promising solution to the loss of a limb with the development of advanced surgical techniques and new, highly effective immunosuppressive agents. METHODS: We retrospectively reviewed the records of 15 hand allotransplantations in 12 patients in China which were performed from September 1999 to May 2008. RESULTS: In total, there were 1 bilateral and 5 unilateral hand transplantations, 3 unilateral and 2 bilateral forearm transplantations, and 1 palm and 1 thumb transplantation. The average age of recipients was 34 ± 11.3 years (range, 19-52 years). At 1-year follow-up, all grafts were viable and with good function. Of the 15 hands transplanted, 8 are currently viable (mean follow-up, 52 ± 36.3 months; range, 16-112 months), including all 3 bilateral cases. Reasons for graft failure were rejection and failure of compliance with immunosuppressive therapy. CONCLUSIONS: Long-term survival of hand transplantation with appropriate immunosuppression is feasible, and satisfactory functional results have been achieved. Careful pretransplant psychologic and social evaluation, consideration of the financial burden of long-term immunosuppressive medications, and close multispecialty collaboration is critical for good outcomes. Limb rejection was related with immunosuppression use. Further study and experience is required before hand allotransplantation can become a generally recommended treatment.

[Effect of platelet-rich plasma injection on early healing of Achilles tendon rupture in rats].

OBJECTIVE: Platelet-rich plasma (PRP) can promote wound healing. To observe the effect of PRP injection on the early healing of rat's Achilles tendon rupture so as to provide the experimental basis for clinical practice. METHODS: Forty-six Sprague Dawley rats were included in this experiment, female or male and weighing 190-240 g. PRP and platelet-poor plasma (PPP) were prepared from the heart arterial blood of 10 rats; other 36 rats were made the models of Achilles tendon rupture, and were randomly divided into 3 groups (control group, PPP group, and PRP group), 12 rats for each group. In PPP and PRP groups, PPP and PRP of 100 microL were injected around the tendons once a week, respectively; in the control group, nothing was injected. The tendon tissue sample was harvested at 1, 2, 3, and 4 weeks after operation for morphology, histology, and immunohistochemistry observations. The content of collagen type I fibers also was measured. Specimens of each group were obtained for biomechanical test at 4 weeks. RESULTS: All the animals survived till the end of the experiment. Tendon edema gradually decreased and sliding improved with time. The tendon adhesion increased steadily from 1 week to 3 weeks postoperatively, and it was relieved at 4 weeks in 3 groups. There was no significant difference in the grading of tendon adhesion among 3 groups at 1 week and at 4 weeks (P > 0.05), respectively. The inflammatory cell infiltration, angiogenesis, and collagen fibers were more in PRP group than in PPP group and control group at 1 week; with time, inflammatory cell infiltration and angiogenesis gradually decreased. Positive staining of collagen type I fibers was observed at 1-4 weeks postoperatively in 3 groups. The positive density of collagen type I fibers in group PRP was significantly higher than that in control group and PPP group at 1, 2, and 3 weeks (P < 0.05), but no significant difference was found among 3 groups at 4 weeks (P > 0.05). The biomechanical tests showed that there was no significant difference in the maximal gliding excursion among 3 groups at 4 weeks postoperatively (P > 0.05); the elasticity modulus and the ultimate tensile strength of PRP group were significantly higher than those of control group and PPP group at 4 weeks (P < 0.05). CONCLUSION: PRP injection can improve the healing of Achilles tendon in early repair of rat's Achilles tendon rupture.

[Establishment of a goat model of tibial bone hole defect suitable for repair using injectable bone materials].

OBJECTIVE: To prepare a goat model of tibial bone hole defect suitable for studies of bone defect repair using tissue-engineered injectable bone materials. METHODS: A circular hole bone defect 1.2 cm in diameter was induced below the tibial medial plateau of the goat. X-ray, histological inspection, and image analysis were carried out to evaluate the validity of the model in simulating limb bone defect for the study of tissue-engineered injectable bone materials. RESULTS: At 4 and 8 weeks after the operation, neither X-ray nor histological examination showed obvious bone tissues in the bone defect. Image analysis showed a area of new bone tissue formation of (8.79 - or + 3.63)% in the total defect area at 4 weeks, which increased to (15.41 - or + 4.21)% at 8 weeks. CONCLUSION: The goat model of tibial bone hole defect established in this study is suitable for studying the ability of injectable bone materials for repairing limb bone defect, and offers a simple and reliable means to simulate the local condition of bone regeneration and mechanical environment of bone defect in the limbs.

[Design and preliminary clinical application of a new digitalized navigation template for fixation of the inferior tibiofibular joint].

OBJECTIVE: To design and prepare a new digitalized navigation template for fixation of inferior tibiofibular joint using three-dimensional reconstruction and reverse engineering techniques. METHODS: Five patients with inferior tibiofibular joint rupture without fibula fracture underwent three-dimensional CT scanning of the lower limbs. The image data were transferred into Mimics software, and after reconstruction of the three-dimensional models of inferior tibiofibular joint rupture and saving in .stl format, the three-dimensional models were imported into Imageware10.0 software to determine the three-dimensional plane of reference. The location of the optimal pedicle channel was defined using reverse engineering and AO internal fixation principle. The template was designed according to the anatomic features of the fibular surface, and the optimal pedicle channel and the template were overlapped as the navigational template, which was manufactured by rapid prototyping. The inferior tibiofibular joint was reduced and the template was placed distally on the external fibula, and the location for screw insertion was defined by the navigation template. RESULTS AND CONCLUSION: The digitalized model of the inferior tibiofibular joint was established. The navigation template manufactured offered good compatibility and was applied successfully for fixation of the inferior tibiofibular joint. This approach provides a new means for fixation of ruptured inferior tibiofibular joint using the reverse engineering and digitized 3-dimensional reconstruction techniques.

[Dynamic monitoring of sIL-2R, IL-2 and IL-6 levels of serum after double-hand transplantation].

OBJECTIVE: To investigate the value of monitoring cytokines in serum after human double-hand allotransplantation. METHODS: We detected solube interleukin-2 receptor(sIL-2R), interleukin-2(IL-2) and interleukin-6(IL-6) levels of the serum in the patient who were performed double-hand transplantation with immunosuppressive from September 2000 to November 2000, and in 15 healthy volunteers by using enzyme linked immunosorbent assays. RESULTS: The IL-2 and IL-6 levels decreased quickly during inducing period (P < 0.05), and then increased gradually. While the sIL-2R level increased. The IL-2 level was lower(P < 0.05), the IL-6 and sIL-2R levels were slightly higher when compared with those before transplantation. There was significant difference in the IL-2 level between the patient and healthy volunteers both before and after transplantation, in the IL-6 level between the patient and healthy volunteers during inducing period. CONCLUSION: IL-2, IL-6 and sIL-2R levels of serum may be useful in diagnosis of rejection, prevention of opportunistic infection, tumor and graft versus host disease.

Monitoring of T lymphocyte subset during ATG induction therapy in hand allograft with report of 3 cases.

OBJECTIVE: To investigate the significance of T lymphocyte subset determination during antithymocyte globulin (ATG) induction therapy in reducing the total drug dose, incidence of complications and cost of treatment in hand allograft. METHODS: The changes in peripheral blood T lymphocyte subsets (CD3+, CD4+, CD8+, and CD28) were determined by flow cytometry in 3 cases of hand allograft who received ATG treatment. RESULTS: Flow cytometry showed that the percentages of CD3+, CD4+, and CD8+ T lymphocytes, along with the ratio of CD4/CD8, decreased significantly during ATG induction therapy, and the results were consistent in the 3 cases. Long-term continuous changes of peripheral blood lymphocytes were observed after antithymocyte globulin induction therapy. CONCLUSION: The understanding of the immunological state of the patient with hand allograft after ATG induction therapy by monitoring T lymphocyte subsets may allow adjustment of the total dose of the drugs administered and help prevent the occurrence of complications.