Human osteoprogenitor cells obtained from traumatic heterotopic ossification samples showed enhanced osteogenic differentiation potential and ERK/hedgehog signaling than that from normal bone.

Traumatic heterotopic ossification (HO) refers to the abnormal ectopic osteogenesis following trauma, causing limb dysfunction and seriously lowering the life quality of patients. Aberrant osteogenic behavior of progenitor cells that ectopically accumulated within the soft tissues are believed to be responsible for HO formation. However, the detailed mechanism still remained to be clarified. Here in this study, we successfully isolated osteoprogenitors from human heterotopic ossification tissues (HO-ops) and identified their stemness and multi-directional differentiation potential. Using alkaline phosphatase staining together with alizarin red staining, we confirmed that the HO-ops in the heterotopic ossified tissues gained greater osteogenic potential than the normal human bone marrow mesenchymal stem cells (HBMSCs). RT-qPCR also indicated that HO-ops obtained more gene transcriptions of critical osteogenic determinators than HBMSCs. In addition, through Western blot, we proved that ERK signaling pathway and hedgehog signaling pathway were significantly activated in the HO-ops. When U0126 and cyclopamine were used to inhibit ERK signaling and hedgehog signaling respectively, the osteogenic potential of HO-ops decreased significantly. The hedgehog signaling and ERK signaling also showed cross-talk in HO-ops during osteogenic differentiation in HO-ops during osteogenic differentiation. The elevated ERK signaling and hedgehog signaling were further confirmed in the human traumatic HO sample sections by immunohistochemical staining. In sum, our results showed that the activation of ERK and hedgehog signaling pathway jointly enhanced the osteogenic potential of HO-ops to induce the formation of traumatic HO, which provides novel insights into the molecular basis of HO formation and offers promising targets for future therapeutic strategy.

Adjusted association between type 2 immunity and low risk thyroid nodules: a retrospective cohort study.

BACKGROUND: Immune responses, especially type 2 immunity, might be related to the prevalence of thyroid nodules, while the key regulators and potential pathways are remaining largely unknown. In addition, the immune status of individuals could be affected by mixed metabolic background. Herein our aim was to investigate the adjusted association between ultrasound-diagnosed low risk thyroid nodules and immune responses, excluding the interference of metabolic effects on immunity. METHODS: We retrospectively enrolled 1764 subjects who underwent a thorough thyroid ultrasound examination. To eliminate the interference of confounders, we used propensity score matching (PSM) to match age, gender, cigarette smoking and alcohol drinking, parameters that are related with metabolic syndrome (MetS). Then the potential effectors of immune responses involved in the laboratorial assays were evaluated. Binary logistic regression analysis was used to assess the independent predictors of thyroid nodules in a multivariate manner. RESULTS: The 1172 subjects were remained after PSM, and differences of demographic background between subjects with and without thyroid nodules were eliminated. Metabolic parameters comprising blood pressure, fasting blood glucose, total cholesterol, triglyceride, high-density lipoprotein, low-density lipoprotein and serum uric acid were shown no significant difference between post-PSM subjects with and without thyroid nodules. Among the biochemistry and hematological parameters, white blood cell count and the positive rate of eosinophil percentage were increased in subjects with thyroid nodules than in those without thyroid nodules. In contrast, the positive rate of basophil percentage was lower in subjects with thyroid nodules than in those without thyroid nodules. In addition, the thyroid function test results showed that subjects with thyroid nodules had higher positive rates of antithyroglobulin antibody (TgAb) and antithyroid peroxidase antibody (TPOAb) than subjects without thyroid nodules. The logistic regression analysis indicated that the positive value of TgAb as well as high level of white blood cell count and BMI could serve as independent risk factors of thyroid nodules. CONCLUSIONS: The type 2 immune responses mediated by increased level of eosinophils, along with positive value of TgAb and TPOAb were associated with the presence of thyroid nodules. In addition, the potential role of basophils in protecting against thyroid nodules and the pathogenesis of immune-metabolic status remains to be elucidated.

Synthesis and evaluation of photo-activatable ß-diarylsydnone-l-alanines for fluorogenic photo-click cyclization of peptides.

Herein, we design and synthesize a series of photoactivatable ß-diarylsydnone-l-alanines (DASAs), which have excellent photo-reactivity with high fluorescence turn-on toward alkenes in a biocompatible environment. The environmental sensing properties of the resulting fluorescent pyrazoline-alanine facilitate its probing capability. By introducing the DASA residue on the side chain of linear peptides, the macrocyclic peptides resulting from the in situ photo-cyclization toward the alkene residue exhibited fluorogenic translocation through live cell membranes.

Discovery of Fluorogenic Diarylsydnone-Alkene Photoligation: Conversion of ortho-Dual-Twisted Diarylsydnones into Planar Pyrazolines.

A small library of diarylsydnones (DASyds) was constructed based on aryl-pairing combinations and subjected to click reaction toward alkenes under photoirradiation with high efficiency. We were able to demonstrate the utility of DASyds for highly fluorescent turn-on ligation targeting the trans-cyclooct-4-en-1-ol moieties on protein.

Multi-layer electrospun membrane mimicking tendon sheath for prevention of tendon adhesions.

Defect of the tendon sheath after tendon injury is a main reason for tendon adhesions, but it is a daunting challenge for the biomimetic substitute of the tendon sheath after injury due to its multi-layer membrane-like structure and complex biologic functions. In this study, a multi-layer membrane with celecoxib-loaded poly(l-lactic acid)-polyethylene glycol (PELA) electrospun fibrous membrane as the outer layer, hyaluronic acid (HA) gel as middle layer, and PELA electrospun fibrous membrane as the inner layer was designed. The anti-adhesion efficacy of this multi-layer membrane was compared with a single-layer use in rabbit flexor digitorum profundus tendon model. The surface morphology showed that both PELA fibers and celecoxib-loaded PELA fibers in multi-layer membrane were uniform in size, randomly arrayed, very porous, and smooth without beads. Multi-layer membrane group had fewer peritendinous adhesions and better gliding than the PELA membrane group and control group in gross and histological observation. The similar mechanical characteristic and collagen expression of tendon repair site in the three groups indicated that the multi-layer membrane did not impair tendon healing. Taken together, our results demonstrated that such a biomimetic multi-layer sheath could be used as a potential strategy in clinics for promoting tendon gliding and preventing adhesion without poor tendon healing.

Optimization of parameters of Yi Zhi Chan Tuina manipulation promotes peripheral circulation.

OBJECTIVE: To explore the most effective parameters of Yi Zhi Chan Tuina manipulation for improving peripheral blood circulation. METHODS: A total of 45 volunteers were recruited from Pudong district in Shanghai, China, from October to December 2010, and randomly divided into nine groups using computer-generated random numbers. Participants received Yi Zhi Chan Tuina manipulation on Chengjin (BL 56) acupoint; each group received a particular combination of manipulation force and treatment time. We used a two-factor, three-level factorial design to examine the effects of force and treatment time on changes in popliteal artery average volume flow, pulsatility index, and vessel diameter to determine the optimal parameter group. Outcomes were assessed at baseline and after Tuina manipulation by interviewers blind to treatment group status. RESULTS: After manipulation, two of the nine groups showed an increase in popliteal artery volume flow. An inter-participants effect test showed that for main effect of time, F = 0.331, P = 0.720; for main effect of force, F = 2.934, P = 0.066; and for the force-time interaction effect, F = 1.072, P = 0.385, indicating no interaction between force and time. However, a pairwise comparison of the three levels of time showed that a treatment time of 10 min was significantly more effective than that of 2 min (P = 0.024). A pairwise comparison of light force, medium force, and heavy force showed a statistically significant effect for medium force (P = 0.035). CONCLUSION: Yi Zhi Chan Tuina manipulation with vertical force of 9.31 N for 10 min is most effective in improving peripheral circulation.

Stability of severely stiff elbows after complete open release: treatment by ligament repair with suture anchors and hinged external fixator.

BACKGROUND: Instability is a crucial issue in severe post-traumatic elbow stiffness during complete-release surgery. This study aimed to evaluate the efficacy of ligament repair using a suture anchor in the operative treatment of severely stiff elbows for which a hinged external fixator was indicated. METHODS: We retrospectively reviewed 46 cases of severely stiff elbows (flexion arc <60°) undergoing open release. During the operation, all 46 elbows were noted to have instability. Suture anchors were applied to restore the ligament if it was impossible to repair the ligament directly, and a hinged external fixator was simultaneously applied to protect the vulnerable ligament and facilitate rehabilitation. No allograft or autograft was used in any of our cases. The stability, arc of motion, Mayo Elbow Performance Score, ulnar nerve symptoms, and radiographs were evaluated. RESULTS: At a mean follow-up of 24.3 months, the postoperative Mayo Elbow Performance Score was 91 points, as compared with 63 points preoperatively. The mean flexion arc improved from 25° to 126°. Three patients presented with moderate elbow instability when the hinged external fixator was removed; however, all of them regained stability by the last follow-up. Furthermore, 7 cases of new-onset nerve palsy were noted; however, all of them resolved with conservative management. None of the patients required secondary surgery for any reason. CONCLUSIONS: Repair of an avulsed collateral ligament with suture anchors and hinged external fixation was effective in restoring functional mobility in patients with severe post-traumatic elbow stiffness after complete release. This could be an option for treating ankylosed, severely or very severely stiff elbows.

Target NF-κB p65 for preventing posttraumatic joint contracture in rats.

RelA/p65 is as a crucial component of the nuclear factor κB (NF-κB) signaling pathway that has a significant impact on various fibrotic diseases. However, its role in the fibrosis of tissues surrounding the joint after traumatic injury remains unclear. In this study, rats were divided into three groups: non-operated control (NC) group, p65-siRNA treated (siRNA-p65) group, and negative siRNA treated (siRNA-neg) group. Then, 10 µL (10 nmol) of p65-siRNA was injected into the joint of the siRNA-p65 group. Meanwhile, 10 µL of negative siRNA was administered to the knee joint of the operated siRNA-neg group for comparison. The rats in the NC group did not receive surgery or drug intervention. After 4 weeks of right knee fixation in each group, X-ray measurements revealed significantly reduced degree of knee flexion contracture following p65-siRNA treatment (siRNA-neg: 77.73° ± 2.799°; siRNA-p65: 105.7° ± 2.629°, p < 0.0001). Histopathological examination revealed that the number of dense fibrous connective tissues decreased following p65-siRNA inhibition. Western blot analysis revealed significantly different expression levels of fibrosis-related proteins between the siRNA-p65 and siRNA-neg groups. Immunohistochemical analysis revealed a reduction in the average number of myofibroblasts in the siRNA-p65 group compared with that in the siRNA-neg group. Thus, intra-articular p65-siRNA injection could attenuate fibroblast activation and fibrosis-related protein production, suppress periarticular tissue fibrosis, and prevent joint contracture by downregulating the NF-κB p65 pathway. Statement of clinical significance: Intra-articular injection of p65-siRNA could reduce myofibroblast proliferation and fibrosis-related protein expression by downregulating the NF-κB p65 pathway, inhibit periarticular tissue fibrosis, and prevent joint adhesion, which represents a potential therapy in the prevention of joint fibrosis following traumatic injury.

Mesoporous Silicon with Strontium-Powered Poly(Lactic-Co-Glycolic acid)/Gelatin-Based Dressings Facilitate Skin Tissue Repair.

Purpose: Functional inorganic nanomaterials (NMs) are widely exploited as bioactive materials and drug depots. The lack of a stable form of application of NMs at the site of skin injury, may impede the removal of the debridement, elevate pH, induce tissue toxicity, and limit their use in skin repair. This necessitates the advent of innovative wound dressings that overcome the above limitations. The overarching objective of this study was to exploit strontium-doped mesoporous silicon particles (PSiSr) to impart multifunctionality to poly(lactic-co-glycolic acid)/gelatin (PG)-based fibrous dressings (PG@PSiSr) for excisional wound management. Methods: Mesoporous silicon particles (PSi) and PSiSr were synthesized using a chemo-synthetic approach. Both PSi and PSiSr were incorporated into PG fibers using electrospinning. A series of structure, morphology, pore size distribution, and cumulative pH studies on the PG@PSi and PG@PSiSr membranes were performed. Cytocompatibility, hemocompatibility, transwell migration, scratch wound healing, and delineated angiogenic properties of these composite dressings were tested in vitro. The biocompatibility of composite dressings in vivo was assessed by a subcutaneous implantation model of rats, while their potential for wound healing was discerned by implantation in a full-thickness excisional defect model of rats. Results: The PG@PSiSr membranes can afford the sustained release of silicon ions (Si4+) and strontium ions (Sr2+) for up to 192 h as well as remarkably promote human umbilical vein endothelial cells (HUVECs) and NIH-3T3 fibroblasts migration. The PG@PSiSr membranes also showed better cytocompatibility, hemocompatibility, and significant formation of tubule-like networks of HUVECs in vitro. Moreover, PG@PSiSr membranes also facilitated the infiltration of host cells and promoted the deposition of collagen while reducing the accumulation of inflammatory cells in a subcutaneous implantation model in rats as assessed for up to day 14. Further evaluation of membranes transplanted in a full-thickness excisional wound model in rats showed rapid wound closure (PG@SiSr vs control, 96.1% vs 71.7%), re-epithelialization, and less inflammatory response alongside skin appendages formation (eg, blood vessels, glands, hair follicles, etc.). Conclusion: To sum up, we successfully fabricated PSiSr particles and prepared PG@PSiSr dressings using electrospinning. The PSiSr-mediated release of therapeutic ions, such as Si4+ and Sr2+, may improve the functionality of PLGA/Gel dressings for an effective wound repair, which may also have implications for the other soft tissue repair disciplines.

Mild synthesis of ultra-bright carbon dots with solvatochromism for rapid lipid droplet monitoring in varied physiological processes.

Lipid droplets (LDs) participating in various cellular activities and are increasingly being emphasized. Fluorescence imaging provides powerful tool for dynamic tracking of LDs, however, most current LDs probes remain inconsistent performance such as low Photoluminescence Quantum Yield (PLQY), poor photostability and tedious washing procedures. Herein, a novel yellow-emissive carbon dot (OT-CD) has been synthesized conveniently with high PLQY up to 90%. Besides, OT-CD exhibits remarkable amphiphilicity and solvatochromic property with lipid-water partition coefficient higher than 2, which is much higher than most LDs probes. These characters enable OT-CD high brightness, stable and wash-free LDs probing, and feasible for in vivo imaging. Then, detailed observation of LDs morphological and polarity variation dynamically in different cellular states were recorded, including ferroptosis and other diseases processes. Furthermore, fast whole imaging of zebrafish and identified LD enrichment in injured liver indicate its further feasibility for in vivo application. In contrast to the reported studies to date, this approach provides a versatile conventional synthesis system for high-performance LDs targeting probes, combing the advantages of easy and high-yield production, as well as robust brightness and stability for long-term imaging, facilitating investigations into organelle interactions and LD-associated diseases.

Prevention of intra-abdominal adhesion by bi-layer electrospun membrane.

The aim of this study was to compare the anti-adhesion efficacy of a bi-layer electrospun fibrous membrane consisting of hyaluronic acid-loaded poly(ε-caprolactone) (PCL) fibrous membrane as the inner layer and PCL fibrous membrane as the outer layer with a single-layer PCL electrospun fibrous membrane in a rat cecum abrasion model. The rat model utilized a cecal abrasion and abdominal wall insult surgical protocol. The bi-layer and PCL membranes were applied between the cecum and the abdominal wall, respectively. Control animals did not receive any treatment. After postoperative day 14, a visual semiquantitative grading scale was used to grade the extent of adhesion. Histological analysis was performed to reveal the features of adhesion tissues. Bi-layer membrane treated animals showed significantly lower adhesion scores than control animals (p < 0.05) and a lower adhesion score compared with the PCL membrane. Histological analysis of the bi-layer membrane treated rat rarely demonstrated tissue adhesion while that of the PCL membrane treated rat and control rat showed loose and dense adhesion tissues, respectively. Bi-layer membrane can efficiently prevent adhesion formation in abdominal cavity and showed a significantly decreased adhesion tissue formation compared with the control.

Combined effect of SDF-1 peptide and angiogenic cues in co-axial PLGA/gelatin fibers for cutaneous wound healing in diabetic rats.

Skin regeneration is hindered by poor vascularization, prolonged inflammation, and excessive scar tissue formation, which necessitate newer strategies to simultaneously induce blood vessel regeneration, resolve inflammation, and induce host cell recruitment. Concurrent deployment of multiple biological cues to realize synergistic reparative effects may be an enticing avenue for wound healing. Herein, we simultaneously deployed SDF (stromal cell-derived factor)- 1α, VEGF (vascular endothelial growth factor)-binding peptide (BP), and GLP (glucagon like peptide)- 1 analog, liraglutide (LG) in core/shell poly(L-lactide-co-glycolide)/gelatin fibers to harness their synergistic effects for skin repair in healthy as well as diabetic wound models in rats. Microscopic techniques, such as SEM and TEM revealed fibrous and core/shell type morphology of membranes. Boyden chamber assay and scratch-wound assay displayed significant migration of HUVECs (human umbilical vein endothelial cells) in SDF-1α containing fibers. Subcutaneous implantation of membranes revealed higher cellular infiltration in SDF-1α loaded fibers, especially, those which were co-loaded with LG or BP. Implantation of membranes in an excisional wound model in healthy rats further showed significant and rapid wound closure in dual cues loaded groups as compared to control or single cue loaded groups. Similarly, the implantation of dressings in type 2 diabetes rat model revealed fast healing, skin appendages regeneration, and blood vessel regeneration in dual cues loaded fibers (SDF-1α/LG, SDF-1α/BP). Taken together, core/shell type fibers containing bioactive peptides significantly promoted wound repair in healthy as well as diabetic wound models in rats.

Electrophysiological, biomechanical, and finite element analysis study of sacral nerve injury caused by sacral fracture.

Background: We aimed to study the mechanism of sacral nerve injury caused by sacral fractures and the relationship between nerve decompression and nerve function. Methods: First, we observed the anatomical features of lumbosacral nerve root region in Sprague-Dawley rats. Next, the rats were divided into the sham, 10 g, 30 g, and 60 g groups for electrophysiological studies on nerve root constriction injury. Then we studied the biomechanical properties of rat nerve roots, lumbosacral trunk, and sacrum. Finally, we established a finite element analysis model of sacral nerve roots injury in rats and determined the correlation between sacral deformation and the degree of sacral nerve roots injury. Result: Anatomical study showed L5 constitutes sciatic nerve, the length of the L5 nerve root is 3.67 ± 0.15 mm, which is suitable for electrophysiological research on nerve root compression injury. After a series of electrophysiological study of L5 nerve roots, our results showed that nerve root function was almost unaffected at a low degree of compression (10 g). Nerve root function loss began at 30 g compression, and was severe at 60 g compression. The degree of neurological loss was therefore positively correlated with the degree of compression. Combining biomechanical testing of the lumbosacral nerve roots, finite element analysis and neuroelectrophysiological research, we concluded when the sacral foramina deformation is >22.94%, the sacral nerves lose function. When the compression exceeds 33.16%, early recovery of nerve function is difficult even after decompression. Conclusion: In this study, we found that the neurological loss was positively correlated with the degree of compression. After early decompression, nerve root function recovery is possible after moderate compression; however, in severe compression group, the nerve function would not recover. Furthermore, FEA was used to simulate nerve compression during sacral fracture, as well as calculate force loading on nerve with different deformation rates. The relationship between sacral fractures and neurological loss can be analyzed in combination with neurophysiological test results.

Water-Involved Ring-Opening of 4-Phenyl-1,2,4-triazoline-3,5-dione for "Photo-Clicked" Access to Carbamoyl Formazan Photoswitches In Situ.

Cyclic azodicarbonyl derivatives, particularly 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD), commonly serve as arenophile, dienophile, enophile and electrophile. Perplexed by its instability in aqueous environment, there are few studies focused on the transient intermediate produced by hydrolysis of PTAD to achieve synthetic significance. Herein, we describe a "photo-click" method that involves nitrile imine (NI) from diarylsydnone to capture the diazenecarbonyl-phenyl-carbamic acid (DACPA) generated by water-promoted ring-opening of PTAD. DFT calculation reveal that H-bonding interactions between PTAD and water are vital to form DACPA which exhibited an umpolung effect during ligation by nature bond orbit (NBO) analysis. The ultra-fast ligation resulted in carbamoyl formazans, as a unique Z↔E photo-switchable linker on target molecules, including peptide and drugs, with excellent anti-fatigue performance. This strategy is showcased to construct highly functionalized carbamoyl formazans in situ for photo-pharmacology and material studies, which also expands the chemistry of PTAD in aqueous media.

One-step synthesis of multifunctional nanoparticles for CT/PA imaging guided breast cancer photothermal therapy.

Advances in nanotheranostics have promoted the development of precision medicine, which has great potential as a weapon for clinical diagnosis and therapy of tumors. However, the combination of three functional principle components (imaging probes, therapeutic agents and surface coating) in traditional theranostic system is difficult to be achieved in only one step, while undergoing multiple synthesis procedures, time-consuming process and unknown toxicity. Herein, we fabricated iodinated polyaniline (LC@I-PANi) nanoparticles via a facile one-step synthesis approach integrating chemical oxidative polymerization and iodine-doping process for computed tomography (CT) imaging and photoacoustic (PA) imaging-guided photothermal therapy (PTT). Iodic acid (HIO3) as an oxidant induces chemical oxidation polymerization of aniline monomers. Meanwhile, iodine is incorporated into the polyaniline structural units in the process of polymerization to obtain LC@I-PANi nanoparticles. Moreover, thel-cysteine (LC) has an effect on diameter of LC@I-PANi nanoparticles, which enables nanoparticles have size-controlled spherical morphology and good colloidal stability. The hemolysis assay and cytotoxicity assessment verified the good biocompatibility of LC@I-PANi. Moreover, our LC@I-PANi nanoparticles could not only exhibit appealing PTT efficiency, but also achieve excellent CT/PA dual-mode imaging effect. The histological evaluations suggested the negligible toxicity of LC@I-PANi in vivo. This is the first time to our knowledge that multifunctional LC@I-PANi nanoparticles were prepared by an ingenious one-step method. This work not only highlights a one-step strategy that simplified the complex synthesis of LC@I-PANi nanoparticles, but also provides insight for further biomedical application of "all-in-one" theranostic agent.

Dual Colorimetric/Fluorometric Double-Throw pH-Switches: The Dimroth Rearrangement of N,9-Diaryl 8-Azaadenines.

A library of 12N,9-Diaryl 2-methyl-8-azaadenine (DAMA) compounds was designed and constructed through an aryl-pairing combination strategy for identifying a nucleobase-containing molecular switch that functions by the pH-regulated Dimroth rearrangement. By utilizing 2D thin-layer chromatography/mass spectrometry (2D-TLC-MS), the DAMA compounds were easily screened to identify which compounds could be used as molecular switches. The pH-switching ability of the DAMA was achieved by incorporating the acridine group as the key structural unit, as well as dual-modal colorimetric/fluorometric on/off properties as the probe functions. The real-time tracing of the switching process clearly indicated that the paired aromatics on both terminals of the DAMA molecule play a key role in tuning the switching kinetics.

Photo-accelerated "click" reaction between diarylsydnones and ring-strained alkynes for bioorthogonal ligation.

We constructed a library of diarylsydnone (DASyd) candidates in search of a photoclickable reaction toward alkynes, enabling an ultra-accelerated reactivity, while suppressing the background cycloaddition in the dark. The in vitro and in vivo protein labelling experiments revealed that the photo-accelerated DASyd-alkyne cycloaddition exhibits robust selectivity.

Possible mechanism underlying analgesic effect of Tuina in rats may involve piezo mechanosensitive channels within dorsal root ganglia axon.

OBJECTIVE: To demonstrate the analgesic effect of Tuina mainly from mechanically sensitive ion channels in peripheral myelinated nerve fibers. METHODS: A total of 40 healthy and pathogen-free adult male Sprague-Dawley rats were used in the study [weight: (220.0 ¡À 1.4) g, Shanghai Slac Laboratory Animal Co., Ltd., Shanghai, China; license No. Shanghai ICP 05033115]. The rats were housed in cages with free access to water and food in a temperature-controlled room [(22 ¡À 1) ¡æ and 12-h/12-h light-dark cycle. Thirty-two rats were randomly divided into five groups: naive, sham, chronic compression of dorsal root ganglion (CCD), Tuina (7 d) and Tuina (21 d). CCD rat model was established via unilateral DRG compression by ""L"" liked steel bar. Chinese Tuina treatment was accepted once per day. Behavior monitoring of paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) were tested. The expression of Piezo1 and Piezo2 in myelinated nerve fiber were analyzed by immunohistochemistry and Western-blotting. RESULTS: There was a high expression of Piezo2 and a low expression of Piezo1 in the naive and CCD groups. In contrast, the expression of Piezo2 was down regulated and Piezo1 was increased after a period of Tuina. There was significant difference (P¡Ü0.05) between the groups. CONCLUSION: Our findings suggest that Tuina therapy can increase the expression of Piezo2 and decrease the expression of Piezo1 in the test rats. The different changes in the expressions of Piezo1 and Piezo2 may play an important role in alleviating CCD-induced allodynia and hyperalgesia.

RelA/p65 inhibition prevents tendon adhesion by modulating inflammation, cell proliferation, and apoptosis.

Peritendinous tissue fibrosis which leads to poor tendon function is a worldwide clinical problem; however, its mechanism remains unclear. Transcription factor RelA/p65, an important subunit in the NF-κB complex, is known to have a critical role in many fibrotic diseases. Here, we show that RelA/p65 functions as a core fibrogenic regulator in tendon adhesion and that its inhibition exerts an anti-fibrogenic effect on peritendinous adhesion. We detected the upregulation of the NF-κB pathway in human tendon adhesion using a gene chip microarray assay and revealed the overexpression of p65 and extracellular matrix (ECM) proteins Collagen I, Collagen III, and α-smooth muscle actin (α-SMA) in human fibrotic tissues by immunohistochemistry and western blotting. We also found that in a rat model of tendon injury, p65 expression correlated with tendon adhesion, whereas its inhibition by small interfering (si)RNA prevented fibrous tissue formation and inflammatory reaction as evidenced by macroscopic, biomechanical, histological, immunohistochemical, and western blotting analyses. Furthermore, in cultured fibroblasts, p65-siRNA, p65-specific inhibitor, Helenalin and JSH23 suppressed cell proliferation and promoted apoptosis, whereas inhibiting the mRNA and protein expression of ECM components and cyclo-oxygenase-2, an inflammatory factor involved in tendon adhesion. Our findings indicate that p65 has a critical role in peritendinous tissue fibrosis and suggest that p65 knockdown may be a promising therapeutic approach to prevent tendon adhesion.

The use of SHP-2 gene transduced bone marrow mesenchymal stem cells to promote osteogenic differentiation and bone defect repair in rat.

Bone tissue engineering is a promising approach for bone regeneration, in which growth factors play an important role. The tyrosine phosphatase Src-homology region 2-containing protein tyrosine phosphatase 2 (SHP2), encoded by the PTPN11 gene, is essential for the differentiation, proliferation and metabolism of osteoblasts. However, SHP-2 has never been systematically studied for its effect in osteogenesis. We predicted that overexpression of SHP-2 could promote bone marrow-derived mesenchymal stem cell (BMSC)osteogenic differentiation and SHP-2 transduced BMSCs could enhance new bone formation, determined using the following study groups: (1) BMSCs transduced with SHP-2 and induced with osteoblast-inducing liquid (BMSCs/SHP-2/OL); (2) BMSCs transduced with SHP-2 (BMSCs/-SHP-2); (3) BMSCs induced with osteoblast-inducing liquid (BMSCs/OL) and (4) pure BMSCs. Cells were assessed for osteogenic differentiation by quantitative real-time polymerase chain reaction analysis, western blot analysis, alkaline phosphatase activity and alizarin red S staining. For in vivo assessment, cells were combined with beta-tricalcium phosphate scaffolds and transplanted into rat calvarial defects for 8 weeks. Following euthanasia, skull samples were explanted for osteogenic evaluation, including micro-computed tomography measurement, histology and immunohistochemistry staining. SHP-2 and upregulation of its gene promoted BMSC osteogenic differentiation and therefore represents a potential new therapeutic approach to bone repair. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1871-1881, 2016.