Free Chimeric Superficial Circumflex Iliac Artery Perforator Flap in Reconstructing the Distal Complex Extensor Tendon Injury.

SUMMARY: The distal complex extensor tendon injury, presenting as traumatic skin, zones 1 and 2 of extensor pollicis longus and extensor hallucis longus, and bony insertion loss, represents a challenging issue and requires a well-vascularized skin paddle, tendinous graft, and insertional reconstruction. Guided by the all-in-one-step reconstruction rule, the chimeric superficial circumflex iliac artery perforator (SCIAP) flap, generally considered as a promising multiple-type tissue provider (eg, vascularized skin paddle, fascia, iliac flap), can fulfill the reconstructive demands and has an edge over the two-stage countermeasure. The authors adopted tripartite SCIAP flaps to reconstruct distal complex thumb or toe injuries in eight cases (six thumbs and two halluces), all of which were reattached with vascularized fascia lata-iliac crest conjunctions using a pull-out technique. All SCIAP flaps survived uneventfully without donor-site complications. The remodeled interphalangeal joints regained nearly normal radiologic manifestation. The chimeric SCIAP flap may be a promising technique for distal complex extensor tendon injury; providing vascularized skin paddle and fascia lata-iliac crest graft, it also qualifies for the all-in-one-stage reconstruction concept. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, IV.

Combination of chemically modified SDF-1α mRNA and small skin improves wound healing in diabetic rats with full-thickness skin defects.

OBJECTIVES: Diabetes mellitus is associated with refractory wound healing, yet current therapies are insufficient to accelerate the process of healing. Recent studies have indicated chemically modified mRNA (modRNA) as a promising therapeutic intervention. The present study aimed to explore the efficacy of small skin engineered to express modified mRNAs encoding the stromal cell-derived factor-1α (SDF-1α) facilitating wound healing in a full-thickness skin defect rat model. This study, devised therapeutic strategies for diabetic wounds by pre-treating small skin with SDF-1α modRNA. MATERIALS AND METHODS: The in vitro transfection efficiency was evaluated using fluorescence microscopy and the content of SDF-1α in the medium was determined using ELISA after the transfection of SDF-1α into the small skin. To evaluate the effect of SDF-1α modRNA and transplantation of the small skin cells on wound healing, an in vivo full-thickness skin defect rat model was assessed. RESULTS: The results revealed that a modRNA carrying SDF-1α provided potent wound healing in the small skin lesions reducing reduced scar thickness and greater angiogenesis (CD31) in the subcutaneous layer. The SDF-1α cytokines were significantly secreted by the small skin after transfection in vitro. CONCLUSIONS: This study demonstrated the benefits of employing small skin combined with SDF-1α modRNA in enhancing wound healing in diabetic rats having full-thickness skin defects.

Regulating mitochondrial homeostasis and inhibiting inflammatory responses through Celastrol.

Background: The high morbidity and mortality rate of coronary heart disease poses a serious threat to human health. Atherosclerosis, a chronic inflammation of the blood vessel wall, is a significant pathological process leading to coronary heart disease. Macrophage inflammation plays a crucial role in the occurrence and development of atherosclerosis. Methods: Macrophage inflammation model was constructed by lipopolysaccharide (LPS), and macrophages were treated with Celastrol at different concentrations (0, 0.1, 1, 10, 100 ng/mL) and different time points (0, 1, 3, 6, 12 h). Real-time quantitative PCR (qPCR) and Western Blot were used to detect the expression of Nur77 mRNA and protein. Macrophages were then pretreated with 100 nmol/L tripterine for 40min and co-cultured with 100 ng/mL LPS. The expression levels of inflammatory factors and chemokines, phosphorylation of phospho-dynamin-related protein 1 (p-Drp1) at Ser637 and expression of mitochondrial fusion protein mitochondrial fusion protein mitofusin-2 (Mfn2) were detected by qPCR, Western blot and ELISA, respectively. The changes of mitochondrial membrane potential were detected by JC-1 probe. Results: 100 nmol/L Celastrol can significantly inhibit LPS-induced inflammatory responses and down-regulate the expression levels of cytokines such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX2), tumor necrosis factor-α (TNF-α), chemokines (CCL-2, and CXCL-10), as well as chemokines. And Celastrol could regulate mitochondrial fission and fusion by promoting the phosphorylation of the Drp1 at the Ser637 site, thereby inhibiting mitochondrial fission. At the same time, by up-regulating the level of the Mfn2, Celastrol also promoted mitochondrial fusion. In addition, we found that the nuclear factor-k-gene binding (NF-κB), extracellular signal-regulated kinase 1/2 (ERK1/2), and p38 signaling pathways aided the drug's anti-inflammatory effects. We also explored the relationship between Celastrol and the nuclear receptor Nur77 and found that it could up-regulate the expression of Nur77. Conclusions: Our study found that Celastrol could reduce inflammation by regulating Drp1 dependent mitochondrial fission and fusion, as well as the ERK1/2, p38, NF-κB signaling pathways. This finding provides a strong direction for the development of new anti-inflammatory drugs for atherosclerosis.

Protective Functions of Liver X Receptor α in Established Vulnerable Plaques: Involvement of Regulating Endoplasmic Reticulum-Mediated Macrophage Apoptosis and Efferocytosis.

Background Liver X receptor (LXR) belongs to the metabolic nuclear receptor superfamily, which plays a critical regulatory role in vascular physiology/pathology. However, effects of systemic LXR activation on established vulnerable plaques and the potential isotype-specific role involved remain unclear. Methods and Results The 8-week-old male apolipoprotein E-/- mice went through carotid branch ligation and renal artery constriction, combined with a high-fat diet. Plaques in the left carotid artery acquired vulnerable features 4 weeks later, confirmed by magnetic resonance imaging scans and histological analysis. From that time on, mice were injected intraperitoneally daily with PBS or GW3965 (10 mg/kg per day) for an additional 4 weeks. Treatment with LXR agonists reduced the lesion volume by 52.61%, compared with the vehicle group. More important, a profile of less intraplaque hemorrhage detection and necrotic core formation was found. These actions collectively attenuated the incidence of plaque rupture. Mechanistically, reduced lesional apoptosis, enhanced efferocytosis, and alleviated endoplasmic reticulum stress are involved in the process. Furthermore, genetic ablation of LXRα, but not LXRß, blunted the protective effects of LXR on the endoplasmic reticulum stress-elicited C/EBP-homologous protein pathway in peritoneal macrophages. In concert with the LXRα-predominant role in vitro, activated LXR failed to stabilize vulnerable plaques and correct the acquired cellular anomalies in LXRα-/- apolipoprotein E-/- mice. Conclusions Our results revealed that LXRα mediates the capacity of LXR activation to stabilize vulnerable plaques and prevent plaque rupture via amelioration of macrophage endoplasmic reticulum stress, lesional apoptosis, and defective efferocytosis. These findings might expand the application scenarios of LXR therapeutics for atherosclerosis.

Hsa_circ_0091581 promotes glioma progression by regulating RMI1 via sponging miR-1243-5p.

Glioma is a pervasive malignancy and the main cause of cancer-related deaths worldwide. Circular RNA is an important subject of cancer research, and its role and function in glioma are poorly understood. This study demonstrated that hsa_circ_0091581 is upregulated in glioma tissues and cells. The results of the CCK-8, EdU, and transwell assays indicated that hsa_circ_0091581 promotes proliferation, migration, and invasion of glioma cells. The results of the luciferase reporter and RNA immunoprecipitation assays indicated that the mechanism of the effects of hsa_circ_0091581 on glioma cells involves sponging miR-1243-5p to regulate RMI1. The results of the rescue experiments indicated that hsa_circ_0091581 regulates proliferation, migration, and invasion of glioma cells by targeting RMI1 in a miR-1243-5p dependent manner. The results of the nude mice xenograft assays showed that knockdown of hsa_circ_0091581 inhibits glioma growth in vivo. Thus, our study determined the role of hsa_circ_0091581/miR-1243-5p/RMI1 in glioma and suggests that this axis may be a novel therapeutic target in glioma.

Ginkgolide B improves multiterritory perforator flap survival by inhibiting endoplasmic reticulum stress and oxidative stress.

BACKGROUND: The therapeutics used to promote perforator flap survival function induces vascular regeneration and inhibit apoptosis. The present study aimed to explore the potential mechanism of the angiogenesis effects of Ginkgolide B (GB) in perforator flaps. Methods: A total of 72 rats were divided into three groups and treated with saline, GB, or GB + tunicamycin (TM; ER stress activator) for seven consecutive days, respectively. Apoptosis was assayed by determining the Bax/Bcl-2 ratio and caspase-3 level. Endoplasmic reticulum (ER) stress markers (CHOP, GRP78, and caspase-12) were detected by Western blot analysis. Oxidative stress was assessed by measuring the superoxide dismutase activity (SOD) and malondialdehyde (MDA), heme oxygenase-1(HO-1), and nuclear factor erythroid-2-related factor 2 (Nrf2) mRNA levels in the flaps. The percentage flap survival area and blood flow were assessed on postoperative day (POD) 7. Angiogenesis was visualized by hematoxylin and eosin and CD34 staining on POD 7. Results: GB increased the survival of perforator flaps, the flap survival area of GB, GB + TM, and control groups was 90.83 ± 1.93%, 70.93 ± 4.13%, and 62.97 ± 6.50%. GB decreased the Bax/Bcl-2 ratio and caspase-3 level. ER stress-related proteins were downregulated by GB. GB also decreased the MDA level and increased SOD activity, HO-1 and Nrf2 mRNA levels in the flaps. Further, GB induced regeneration of vascular vessels in comparison with saline or GB + TM. Conclusions: GB increased angiogenesis and alleviated oxidative stress by inhibiting ER stress, which increased the survival of perforator flaps. In contrast, GB + TM alleviated angiogenesis and induced oxidative stress by activating ER stress and decreasing the survival of perforator flaps.

Macrophage autophagy regulates mitochondria-mediated apoptosis and inhibits necrotic core formation in vulnerable plaques.

The vulnerable plaque is a key distinguishing feature of atherosclerotic lesions that can cause acute atherothrombotic vascular disease. This study was designed to explore the effect of autophagy on mitochondria-mediated macrophage apoptosis and vulnerable plaques. Here, we generated the mouse model of vulnerable carotid plaque in ApoE-/- mice. Application of ApoE-/- mice with rapamycin (an autophagy inducer) inhibited necrotic core formation in vulnerable plaques by decreasing macrophage apoptosis. However, 3-methyladenine (an autophagy inhibitor) promoted plaque vulnerability through deteriorating these indexes. To further explore the mechanism of autophagy on macrophage apoptosis, we used macrophage apoptosis model in vitro and found that 7-ketocholesterol (7-KC, one of the primary oxysterols in oxLDL) caused macrophage apoptosis with concomitant impairment of mitochondria, characterized by the impairment of mitochondrial ultrastructure, cytochrome c release, mitochondrial potential dissipation, mitochondrial fragmentation, excessive ROS generation and both caspase-9 and caspase-3 activation. Interestingly, such mitochondrial apoptotic responses were ameliorated by autophagy activator, but exacerbated by autophagy inhibitor. Finally, we found that MAPK-NF-κB signalling pathway was involved in autophagy modulation of 7-KC-induced macrophage apoptosis. So, we provide strong evidence for the potential therapeutic benefit of macrophage autophagy in regulating mitochondria-mediated apoptosis and inhibiting necrotic core formation in vulnerable plaques.

The Effect of Leonurine on Multiterritory Perforator Flap Survival in Rats.

BACKGROUND: Leonurine (Leo), a natural active compound of Leonurus cardiaca, has been shown to possess various biological activities. However, it is not known whether Leo promotes perforator flap survival. METHODS: In this study, a perforator flap was outlined in the rat dorsum. The rats that survived surgery were divided randomly to control and Leo groups (n = 36 per group). Flap viability, flap perfusion, and level of protein linked with oxidative stress, cell apoptosis, and angiogenesis were evaluated. RESULTS: Relative to control group, the Leo group showed significantly higher the flap survival percentage (70.5% versus 90.2%, P < 0.05) and blood perfusion (197.1 versus 286.3, P < 0.05). Leo also increased 1.8-fold mean vessel density and upregulated 2.1-fold vascular endothelial growth factor protein expression compared with the control group, both of which indicate increased angiogenesis. Moreover, it significantly inhibited apoptosis by lowering caspase-3 activity. Superoxide dismutase expression was remarkably elevated in Leo group compared with the control group (56.0 versus 43.2 U/mg/protein, P < 0.01), but malondialdehyde quantities were significantly lower in the Leo group compared with control group (41.9 versus 57.5 nmol/mg/protein, P < 0.05). CONCLUSIONS: Leo may serve as an effective drug for improving perforator flap survival in rats via antioxidant and antiapoptotic mechanisms and promotion of angiogenesis.

Inhibiting the PI3K/AKT/NF-κB signal pathway with nobiletin for attenuating the development of osteoarthritis: in vitro and in vivo studies.

Osteoarthritis (OA), an age-related degenerative disease, is characterized by progressive degradation of the articular cartilage. There is increasing evidence that nobiletin (NOB) exerts special biological functions in a variety of diseases. However, whether it protects against OA remains unknown. In this study, we investigated the anti-inflammatory and chondroprotective effects of NOB on IL-1ß-induced human OA chondrocytes and in the surgical DMM mice OA models. In vitro, NOB treatment completely suppressed the overproduction of pro-inflammatory mediators, including PGE2, NO, COX-2, iNOS, TNF-α and IL-6 in IL-1ß-induced human OA chondrocytes. Moreover, NOB exerted a potent inhibitory effect on the expression of MMP-13 and ADAMTS-5 as well as the degradation of aggrecan and collagen-II, which leads to the degradation of the extracellular matrix. Furthermore, NOB dramatically suppressed the IL-1ß-stimulated phosphorylation of PI3K/Akt and activation of NF-κB in human OA chondrocytes. In addition, treatment with NOB not only prevented the destruction of cartilage and the thickening of subchondral bone but also relieved synovitis in mice OA models. In conclusion, our study suggests that NOB holds novel therapeutic potential for the treatment of OA.

Analysis of risk factors for loss of reduction after acromioclavicular joint dislocation treated with the suture-button.

BACKGROUND: The most commonly reported complication after acromioclavicular (AC) joint dislocation treated with the Suture-button is loss of reduction. Loss of reduction is a major factor influencing the patient's joint function and subjective satisfaction. The objective of this study is to analyze the risk factors causing loss of reduction after AC joint dislocation treated with the Suture-button. METHODS: One hundred and thirty patients with AC joint dislocation who were surgically treated the Suture-button in our hospital from February 2009 to February 2015, were recorded their age, sex, BMI, time from injury to surgery, Rockwood's classification, with or without osteoporosis, double or triple button technique, position of the clavicle tunnel, tunnel diameter, coracoid button position, alignment of the button, acromioclavicular ligament repair or not, different methods of postoperative limb immobilization, and so on. Mean comparisons or chi-square test was used for univariate analysis of the above factors, and then multivariate logistic regression analysis was performed to predict risk factors. RESULTS: Reduction was lost in 23.1% of the patients. Univariate analysis showed that button alignment, double or triple button technique, coracoid button position, position of the clavicle tunnel, acromioclavicular ligament repair or not and osteoporosis had statistically significant association with loss of reduction for AC joint (P = 0.031, 0.034, 0.000, 0.042, 0.047 and 0.000 respectively). Multivariate logistic regression analysis demonstrated that osteoporosis (P = 0.003), position of the clavicle tunnel (P = 0.032) and coracoid button position (P < 0.001) were the risk factors that significantly associated with the loss of reduction after AC joint dislocation treated with the Suture-button. CONCLUSIONS: Clavicle tunnel location using relative ratio method, accurate placement of button plate under coracoid process (inside or outside deviation <20°), various reinforcement operations for patients with osteoporosis are important factors in preventing loss of reduction.

The Feasibility of 3D Printing Technology on the Treatment of Pilon Fracture and Its Effect on Doctor-Patient Communication.

PURPOSE: The aim of this study was to assess the feasibility and effectiveness of the three-dimensional (3D) printing technology in the treatment of Pilon fractures. METHODS: 100 patients with Pilon fractures from March 2013 to December 2016 were enrolled in our study. They were divided randomly into 3D printing group (n = 50) and conventional group (n = 50). The 3D models were used to simulate the surgery and carry out the surgery according to plan in 3D printing group. Operation time, blood loss, fluoroscopy times, fracture union time, and fracture reduction as well as functional outcomes including VAS and AOFAS score and complications were recorded. To examine the feasibility of this approach, we invited surgeons and patients to complete questionnaires. RESULTS: 3D printing group showed significantly shorter operation time, less blood loss volume and fluoroscopy times, higher rate of anatomic reduction and rate of excellent and good outcome than conventional group (P < 0.001, P < 0.001, P < 0.001, P = 0.040, and P = 0.029, resp.). However, no significant difference was observed in complications between the two groups (P = 0.510). Furthermore, the questionnaire suggested that both surgeons and patients got high scores of overall satisfaction with the use of 3D printing models. CONCLUSION: Our study indicated that the use of 3D printing technology to treat Pilon fractures in clinical practice is feasible.

Comparison of the outcomes of pediatric tibial shaft fractures treated by different types of orthopedists: A prospective cohort study.

BACKGROUND: The objective of this study was to compare the outcomes of pediatric tibial shaft fractures treated with titanium elastic nail (TEN) by pediatric orthopedists and non-pediatric orthopedists. MATERIALS AND METHODS: We conducted a prospective cohort study of 90 children of tibial shaft fractures, who were randomized to operative stabilization either by pediatric orthopedists (Group A, 45 cases) or by non-pediatric orthopedists (Group B, 45 cases) from April 2010 to May 2015. Demographic data and clinical characteristics (age, sex, weight, fracture side and type, cause of injury, number of fibula fracture and time from injury to operation) were comparable between the two groups before surgery. Clinical data, complications and functional outcomes between the two groups were recorded. RESULTS: A total of 10 patients were lost to follow-up for various reasons and each group remained 40 cases. The mean follow-up period was 14.9 ±â€¯1.9 months for Group A and 15.3 ±â€¯2.2 months for Group B (P = 0.451). There was no significant difference in length of hospitalization, full weight-bearing time, fracture union time and TEN outcome scores between the two groups (P = 0.917, P = 0.352, P = 0.404, P = 506, respectively). However, Group A exhibited significantly shorter operation duration and less fluoroscopy times than Group B (P < 0.001 and P < 0.001, respectively). Also, there was a trend for patients of Group A to have lower rate of open reduction than Group B (P = 0.019). When comparing the total complications, no significant difference existed between the groups (P = 0.764). CONCLUSIONS: Our results indicated that pediatric tibial shaft fractures treated surgically by pediatric orthopedists offered potential advantages including a shorter operating time, less times of fluoroscopy and a lower rate of open reduction. However, both pediatric and non-pediatric orthopedists could achieve satisfactory clinical results in treatment of these injuries.

Comparison of three surgical fixation methods for dual-bone forearm fractures in older children: A retrospective cohort study.

BACKGROUND: The aim of this study was to compare the outcomes of dual ESIN (D-ESIN) fixation, hybrid fixation, and open reduction and dual plate (d-plate) fixation in the treatment of dual-bone forearm fractures in children aged 10-16 years. MATERIALS AND METHODS: 137 patients with dual-bone forearm fractures (48 patients in the D-ESIN group, 45 patients in the hybrid group, and 44 patients in the d-plate group) were reviewed. Duration of surgery, length of incision, intraoperative blood loss, intraoperative times of fluoroscopy, and duration of postoperative immobilisation were recorded. Radiographic outcomes, functional outcomes, and complication rate were also recorded. RESULTS: Surgeries and incisions were significantly shorter, and less intraoperative blood loss occurred, in the hybrid group than the d-plate group (P < 0.001). The hybrid group was also characterised by less intraoperative fluoroscopy times and shorter duration of postoperative immobilisation compared with the D-ESIN group (P < 0.001). The union rate of the ulna at 3 months postoperatively was higher in the hybrid and d-plate groups than in the D-ESIN group (P = 0.003). The union rate of the radius was similar in all three groups (P = 0.403). No significant difference in the union rate of the radius or ulna was observed among groups at 6 months postoperatively (P = 0.052). The mean union time was notably later in the D-ESIN group than in the hybrid and d-plate groups. However, no significant difference in functional outcome or complication rate was observed among the three groups (P = 0.822 and P = 0.912). CONCLUSION: Hybrid fixation was superior in terms of the duration of surgery, intraoperative use of fluoroscopy, intraoperative blood loss, duration of postoperative immobilisation, delayed union of the ulna, and bone union time. Therefore, hybrid fixation is a safe and effective treatment for dual-bone forearm fractures in children aged 10-16 years.

Comparison of the Conventional Surgery and the Surgery Assisted by 3d Printing Technology in the Treatment of Calcaneal Fractures.

PURPOSE: This study was aimed to compare conventional surgery and surgery assisted by 3D printing technology in the treatment of calcaneal fractures. In addition, we also investigated the effect of 3D printing technology on the communication between doctors and patients. METHODS: we enrolled 75 patients with calcaneal fracture from April 2014 to August 2016. They were divided randomly into two groups: 35 cases of 3D printing group, 40 cases of conventional group. The individual models were used to simulate the surgical procedures and carry out the surgery according to plan in 3D printing group. Operation duration, blood loss volume during the surgery, number of intraoperative fluoroscopy and fracture union time were recorded. The radiographic outcomes Böhler angle, Gissane angle, calcaneal width and calcaneal height and final functional outcomes including VAS and AOFAS score as well as the complications were also evaluated. Besides, we made a simple questionnaire to verify the effectiveness of the 3D-printed model for both doctors and patients. RESULTS: The operation duration, blood loss volume and number of intraoperative fluoroscopy for 3D printing group was 71.4 ± 6.8 minutes, 226.1 ± 22.6 ml and 5.6 ± 1.9 times, and for conventional group was 91.3 ± 11.2 minutes, 288.7 ± 34.8 ml and 8.6 ± 2.7 times respectively. There was statistically significant difference between the conventional group and 3D printing group (p < 0.05). Additionally, 3D printing group achieved significantly better radiographic results than conventional group both postoperatively and at the final follow-up (p < 0.05). However, No significant difference was noted in the final functional outcomes between the two groups. As for complications, there was no significant difference between the two groups. Furthermore, the questionnaire showed that both doctors and patients exhibited high scores of overall satisfaction with the use of a 3D printing model. CONCLUSION: This study suggested the clinical feasibility of 3D printing technology in treatment of calcaneal fractures.

Silibinin protects against osteoarthritis through inhibiting the inflammatory response and cartilage matrix degradation in vitro and in vivo.

Osteoarthritis (OA) is a degenerative joint disease characterized by cartilage degradation and inflammation. Silibinin, a polyphenolic flavonoid derived from fruits and seeds of Silybum marianum, has been reported to possess various potent beneficial biological effects, such as antioxidant, anti-cancer, hepatoprotective and anti-inflammatory activities. However, the anti-inflammatory effects of silibinin on OA have not been reported. This study aimed to assess the effects of silibinin on OA both in vitro and in vivo. In this study, we found that silibinin significantly inhibited the nterleukin-1ß (IL-1ß)-induced production of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α) and IL-6, expression of cyclooxygenase2 (COX-2), inducible nitric oxide synthase (iNOS), matrix metalloproteinase-1 (MMP-1), MMP-3, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) and ADAMTS-5, degradation of aggrecan and collagen-II in human OA chondrocytes. Furthermore, silibinin dramatically suppressed IL-1ß-stimulated phosphatidylinositol 3 kinase/ protein kinase B (PI3K/Akt) phosphorylation and nuclear factor-kappa B (NF-kB) activation in human OA chondrocytes. In addition, treatment of silibinin not only prevented the destruction of cartilage and the thickening of subchondral bone but also relieved synovitis in mice OA models. Also, the immunohistochemistry results showed that silibinin significantly decreased the expression of MMP-13 and ADAMTS-5 and increased the expression of collagen-II and aggrecan in mice OA. Taken together, these results suggest that silibinin may be a potential agent in the treatment of OA.

Osthole Enhances the Therapeutic Efficiency of Stem Cell Transplantation in Neuroendoscopy Caused Traumatic Brain Injury.

Neuroendoscopy processes can cause severe traumatic brain injury. Existing therapeutic methods, such as neural stem cell transplantation and osthole have not been proven effective. Therefore, there is an emerging need on the development of new techniques for the treatment of brain injuries. In this study we propose to combine the above stem cell based methods and then evaluate the efficiency and accuracy of the new method. Mice were randomly divided into four groups: group 1 (brain injury alone); group 2 (osthole); group 3 (stem cell transplantation); and group 4 (osthole combined with stem cell transplantation). We carried out water maze task to exam spatial memory. Immunocytochemistry was used to test the inflammatory condition of each group, and the differentiation of stem cells. To evaluate the condition of the damaged blood brain barrier restore, we detect the Evans blue (EB) extravasation across the blood brain barrier. The result shows that osthole and stem cell transplantation combined therapeutic method has a potent effect on improving the spatial memory. This combined method was more effective on inhibiting inflammation and preventing neuronal degeneration than the single treated ones. In addition, there was a distinct decline of EB extravasation in the combined treatment groups, which was not observed in single treatment groups. Most importantly, the combined usage of osthole and stem cell transplantation provide a better treatment for the traumatic brain injury caused by neuroendoscopy. The collective evidence indicates osthole combined with neural stem cell transplantation is superior than either method alone for the treatment of traumatic brain injury caused by neuroendoscopy.

Fisetin inhibits IL-1ß-induced inflammatory response in human osteoarthritis chondrocytes through activating SIRT1 and attenuates the progression of osteoarthritis in mice.

Osteoarthritis (OA) is a degenerative joint disease characterized by cartilage degradation and inflammation. Fisetin, a polyphenol extracted from fruits and vegetables, has been reported to have anti-inflammatory effects. Our study aimed to investigate the effect of fisetin on OA both in vitro and in vivo. In vitro, chondrocytes were pretreated with fisetin alone or fisetin combined with sirtinol (an inhibitor of SIRT1) for 2h before IL-1ß stimulation. Production of NO, PGE2, TNF-α and IL-6 were evaluated by the Griess reaction and ELISAs. The mRNA (COX-2, iNOS, MMP-3, MMP-13, ADAMTS-5, Sox-9, aggrecan and collagen-II) and protein expression (COX-2, iNOS, MMP-3, MMP-13, ADAMTS-5 and SIRT1) were measured by qRT-PCR and Western blot respectively. Immunofluorescence was used to assess the expression of collagen-II and SIRT1. SIRT1 activity was quantified with SIRT1 fluorometric assay kit. The in vivo effect of fisetin was evaluated by gavage in mice OA models induced by destabilization of the medial meniscus (DMM). We found that fisetin inhibited IL-1ß-induced expression of NO, PGE2, TNF-α, IL-6, COX-2, iNOS, MMP-3, MMP-13, ADAMTS-5. Besides, fisetin remarkably decreased IL-1ß-induced degradation of Sox-9, aggrecan and collagen-II. Furthermore, fisetin significantly inhibited IL-1ß-induced SIRT1 decrease and inactivation. However, the inhibitory effect of fisetin was obvious abolished by sirtinol, suggesting that fisetin exerts anti-inflammatory effects through activating SIRT1. In vivo, fisetin-treated mice exhibited less cartilage destruction and lower OARSI scores. Moreover, fisetin reduced subchondral bone plate thickness and alleviated synovitis. Taken together, these findings indicate that fisetin may be a potential agent in the treatment of OA.

Neurotrophin-3 promotes proliferation and cholinergic neuronal differentiation of bone marrow- derived neural stem cells via notch signaling pathway.

AIMS: Recently, the potential for neural stem cells (NSCs) to be used in the treatment of Alzheimer's disease (AD) has been reported; however, the therapeutic effects are modest by virtue of the low neural differentiation rate. In our study, we transfected bone marrow-derived NSCs (BM-NSCs) with Neurotrophin-3 (NT-3), a superactive neurotrophic factor that promotes neuronal survival, differentiation, and migration of neuronal cells, to investigate the effects of NT-3 gene overexpression on the proliferation and differentiation into cholinergic neuron of BM-NSCs in vitro and its possible molecular mechanism. MAIN METHODS: BM-NSCs were generated from BM mesenchymal cells of adult C57BL/6 mice and cultured in vitro. After transfected with NT-3 gene, immunofluorescence and RT-PCR method were used to determine the ability of BM-NSCs on proliferation and differentiation into cholinergic neuron; Acetylcholine Assay Kit was used for acetylcholine (Ach). RT-PCR and WB analysis were used to characterize mRNA and protein level related to the Notch signaling pathway. KEY FINDINGS: We found that NT-3 can promote the proliferation and differentiation of BM-NSCs into cholinergic neurons and elevate the levels of acetylcholine (ACh) in the supernatant. Furthermore, NT-3 gene overexpression increase the expression of Hes1, decreased the expression of Mash1 and Ngn1 during proliferation of BM-NSCs. Whereas, the expression of Hes1 was down-regulated, and Mash1 and Ngn1 expression were up-regulated during differentiation of BM-NSCs. SIGNIFICANCE: Our findings support the prospect of using NT-3-transduced BM-NSCs in developing therapies for AD due to their equivalent therapeutic potential as subventricular zone-derived NSCs (SVZ-NSCs), greater accessibility, and autogenous attributes.

Arctigenin Treatment Protects against Brain Damage through an Anti-Inflammatory and Anti-Apoptotic Mechanism after Needle Insertion.

UNLABELLED: Convection enhanced delivery (CED) infuses drugs directly into brain tissue. Needle insertion is required and results in a stab wound injury (SWI). Subsequent secondary injury involves the release of inflammatory and apoptotic cytokines, which have dramatic consequences on the integrity of damaged tissue, leading to the evolution of a pericontusional-damaged area minutes to days after in the initial injury. The present study investigated the capacity for arctigenin (ARC) to prevent secondary brain injury and the determination of the underlying mechanism of action in a mouse model of SWI that mimics the process of CED. After CED, mice received a gavage of ARC from 30 min to 14 days. Neurological severity scores (NSS) and wound closure degree were assessed after the injury. Histological analysis and immunocytochemistry were used to evaluated the extent of brain damage and neuroinflammation. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was used to detect universal apoptosis. Enzyme-linked immunosorbent assays (ELISA) was used to test the inflammatory cytokines (tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-10) and lactate dehydrogenase (LDH) content. Gene levels of inflammation (TNF-α, IL-6, and IL-10) and apoptosis (Caspase-3, Bax and Bcl-2) were detected by reverse transcription-polymerase chain reaction (RT-PCR). Using these, we analyzed ARC's efficacy and mechanism of action. RESULTS: ARC treatment improved neurological function by reducing brain water content and hematoma and accelerating wound closure relative to untreated mice. ARC treatment reduced the levels of TNF-α and IL-6 and the number of allograft inflammatory factor (IBA)- and myeloperoxidase (MPO)-positive cells and increased the levels of IL-10. ARC-treated mice had fewer TUNEL+ apoptotic neurons and activated caspase-3-positive neurons surrounding the lesion than controls, indicating increased neuronal survival. CONCLUSIONS: ARC treatment confers neuroprotection of brain tissue through anti-inflammatory and anti-apoptotic effects in a mouse model of SWI. These results suggest a new strategy for promoting neuronal survival and function after CED to improve long-term patient outcome.