Administration of necrostatin-1 ameliorates glucocorticoid-induced osteonecrosis of the femoral head in rats.

Glucocorticoid (GC)-induced osteonecrosis of the femoral head (ONFH) is a serious complication of glucocorticoid treatment and is characterized by dysfunctional bone reconstruction at necrotic sites. Our previous study confirmed the protective potential of necrostatin-1, a selective blocker of necroptosis, in glucocorticoid-induced osteoporosis. In this study, rat models of GC-induced ONFH were established to evaluate the effects of necrostatin-1 on osteonecrotic changes and repair processes. Osteonecrosis was verified by histopathological staining. An analysis of trabecular bone architecture was performed to evaluate osteogenesis in the osteonecrotic zone. Then, necroptotic signaling molecules such as RIP1 and RIP3 were examined by immunohistochemistry. Histopathological observations indicated that necrostatin-1 administration reduced the incidence of osteonecrosis and the osteogenic response in subchondral areas. Additionally, bone histomorphometry demonstrated that necrostatin-1 intervention could restore bone reconstruction in the necrotic zone. The protective mechanism of necrostatin-1 was related to the inhibition of RIP1 and RIP3. Necrostatin-1 administration alleviated GC-induced ONFH in rats by attenuating the formation of necrotic lesions, recovering the function of osteogenesis, and suppressing glucocorticoid-induced osteocytic necroptosis by inhibiting the expression of RIP1 and RIP3.

Effectiveness of abduction brace versus simple sling rehabilitation following rotator cuff repair: systematic review and meta-analyses.

BACKGROUND: To date, no conclusions have been reached regarding the type of brace worn after arthroscopic rotator cuff repair. To this end, a systematic review and meta-analysis of randomized controlled trials (RCTs) were conducted. METHODS: According to the updated guidelines of the Preferred Reporting Items of Systematic Review and Meta-Analysis, all related literature in PubMed, Embase, and Cochrane Central Register of Controlled Trials, from their establishment to March 1, 2022, were searched systematically. Outcome measures included the Constant score, Western Ontario Rotator Cuff (WORC) index, visual analog scale (VAS) score, shoulder joint range of motion (ROM), and failure events of rotator cuff healing. The Cochrane risk of bias tool was used to evaluate the quality of RCT. RESULTS: Two independent reviewers (Chen, Wu) reviewed 275 articles, of which only five met the inclusion criteria, and four were included in the meta-analysis, with a total of 302 patients. The overall risk of bias was high in two RCTs, unclear in one, and low in two. Considering the clinical outcomes, the Constant score (P = .08 mean deviation [MD], 3.06; 95% confidence interval [CI], -0.42 to 6.53), WORC (P = .23; MD, 3.32; 95%CI, -2.15 to 8.79), VAS score (P = .09; MD -1.27; 95%CI, -2.75 to 0.21), ROM (P = .1; MD, 4.75; 95%CI, -0.98 to 10.48), and failure events of rotator cuff healing (P = .78; odds ratio [OR], 0.86; 95%CI, 0.32 to 2.37) did not significantly differ between the abduction brace and simple sling after arthroscopic rotator cuff repair. CONCLUSION: The findings of this systematic review and meta-analysis suggest that wearing abduction braces after rotator cuff repair neither improved the Constant score, VAS, and WORC scores, and ROM of the shoulder joint, nor did it reduce the risk of re-tearing. Therefore, a simple sling may be a better option in terms of cost effectiveness. It is expected that studies with larger and more homogeneous samples will help verify our results.

Effect and Mechanism of Solidified Microstructure on Deformation Behavior, Mechanical Properties, and Residual Stress of Cu-Ni-Si Alloy.

Cu-Ni-Si alloy is the key raw material for the lead frame of large integrated circuits. The disordered grain orientation of alloy billet, high hardening rate, residual stress, and poor surface quality of cold working strips seriously affect its processability. In order to improve the cold-working properties of Cu-Ni-Si alloy, two kinds of C70250 copper alloy strips were produced through hot mold continuous casting (HMCC) and cold mold continuous casting (CMCC) technology. The effects of solidified microstructure on the cold-working deformation behavior, mechanical properties, and residual stress of the alloy were studied. The results show that C70250 copper alloys with columnar grain and equiaxed grain were prepared through HMCC and CMCC. After a 98% reduction in cold rolling, columnar grain strip surface quality was very good, and the elongation was still as high as 3.2%, which is 2.9 times that of equiaxed grain alloy. The residual stress of equiaxed grain strips reached 363 MPa, which is 2.7 times that of columnar grain strips. During the cold rolling process, equiaxed grain strips are prone to cause intersecting plane dislocations, stacking faults, shear bands, and grain breakage during large deformation cold rolling. The columnar grain strip causes parallel plane dislocations, stacking faults, and shearbands. Furthermore, the deformation structure was found to be uniform, and, ultimately, the alloy formed a fibrous structure. Therefore, the elongation and latter distortion of columnar grain strips improved after being put through large deformation cold rolling, which greatly reduced residual stress.

Astragalin mitigates inflammatory osteolysis by negatively modulating osteoclastogenesis via ROS and MAPK signaling pathway.

Inflammatory bone destruction has gradually attracted attention worldwide and has been observed in several kinds of pathological bone diseases, such as osteoarthritis, osteomyelitis, rheumatic arthritis, and other infectious clinical trials in the skeletal system. In this regard, excessive osteoclasts and bone resorption activity participate in osteolytic processes. Thus, negatively modulating osteoclast differentiation and bone erosion has been considered an effective therapeutic strategy to limit the poor progression of inflammatory osteolysis. Astragalin (AST) is a bioactive component of traditional Chinese drugs, such as Rosa agrestis, which presents anti-inflammatory and antioxidant effects. However, it is unclear how AST may play an essential role in regulating the dynamic balance of the bone matrix by affecting osteoclastogenesis. This study found that AST could inhibit osteoclastic formation and bone resorption activity in a dose-dependent manner without cytotoxicity. Administration of AST also inhibited the expression of cathepsin K, c-Fos, NFATc1, and TRAP at different stages of mRNA and protein levels during osteoclastogenesis. Reactive oxygen species (ROS) signalling could also be modulated by treatment with AST during RANKL-induced osteoclast differentiation through the Nrf2-HO1 signalling pathway. Additionally, AST could negatively regulate mitogen-activated protein kinase (MAPK) signalling in this process. In vivo, AST significantly reduced lipopolysaccharide (LPS)-induced bone loss in an osteolytic mouse model. AST might be a promising therapeutic candidate for treating osteolytic bone diseases in the future.

Development of biodegradable Zn-Mn-Li and CaP coatings on Zn-Mn-Li alloys and cytocompatibility evaluation for bone graft.

Zn-based alloys are considered as new kind of potential biodegradable implanted biomaterials recently. The difficulty of metal implanted biomaterials and bone tissue integration seriously affects the applications of metal implanted scaffolds in bone tissue-related fields. Herein, we self-designed Zn0.8Mn and Zn0.8Mn0.1Li alloys and CaP coated Zn0.8Mn and Zn0.8Mn0.1Li alloys, then evaluated the degradation property and cytocompatibility. The results demonstrated that the Zn0.8Mn0.1Li alloys had profoundly modified the degradation property and cytocompatibility, but Zn0.8Mn0.1Li alloys had particularly adverse effects on the surface morphology of osteoblasts. The results furtherly showed that the CaP-coated Zn0.8Mn and Zn0.8Mn0.1Li alloys scaffold had better biocompatibility, which would further guarantee the biosafety of this new kind of biodegradable Zn-based alloys implants for future clinical applications.

Microporous structures on mineralized collagen mediate osteogenesis by modulating the osteo-immune response of macrophages.

It is a new hot pot in tissue engineering and regenerative medicine to study the effects of physicochemical properties of implanted biomaterials on regulating macrophage polarization to promote bone regeneration. In this study, we designed and fabricated mineralized collagen (MC) with different microporous structures via in vitro biomimetic mineralization method. The microporous structures, mechanical properties, shore hardness and water contact angle measurements were tested. Live/dead cell staining, CCK-8 assay, phalloidine staining, staining of focal adhesions were used to detect cell behavior. ELISA, qRT-PCR, ALP, and alizarin red staining (ARS) were performed to appraise osteogenic differentiation and investigated macrophage response and their subsequent effects on the osteogenic differentiation. The results showed that RAW264.7 and MC3T3-E1 cells were able to survive on the MC. MC with the microporous structure of approximately 84 µm and 70%-80% porosity could promote M2 macrophage polarization and increase the expression level of TGF-ß and VEGF. Moreover, the gene expression of the osteogenic markers ALP, COL-1, and OCN increased. Therefore, MC with different microporous structures mediated osteoimmunomodulation in bone regeneration. These data will provide a new idea of biomaterials inducing bone repair and direct the optimal design of novel immune biomaterials, development, and rational usage.

Comparison of time consumption and success rate between CT angiography- and CT perfusion- based imaging assessment strategy for the patients with acute ischemic stroke.

BACKGROUND: Our study aimed to compare the time consumption and success rate between CTA- and CTP- based assessment strategy, and to clarify the risk factors associated with the CTP scan failure. METHODS: Clinical and radiological data of 437 consecutive AIS patients who underwent multiphase CTA or CTP for pre-treatment evaluation were retrospectively enrolled (CTA group, n = 302; CTP group, n = 135). Time consumption and success rate of CTA- and CTP- based assessment strategy were compared using Mann-Whitney U test and Chi-Squared Test. Univariate analysis and receiver operating curve analysis were used to clarify the risk factors, and their performance in predicting the CTP scan failure. RESULTS: Time consumption of CTP scan and reconstruction was significantly longer than that of CTA [775 s vs 263.5 s, P < 0.001]. CTP scan showed significantly higher failure rate than CTA (11% vs 1%, P < 0.001). Severe motion was the most common cause of CTP failure (n = 12, 80%). Baseline National Institute of Health Stroke Scale (NIHSS) score in CTP failure group was significantly higher than that in CTP success group [17 vs 13, P = 0.007]. Baseline NIHSS score of 11 was the optimal threshold value to predict CTP failure with an area under the curve of 0.715, a sensitivity of 86.7%, and a specificity of 45.0%. CONCLUSIONS: CTP- based strategy showed longer time consumption and higher failure rate than CTA- based strategy. High baseline NIHSS score was significantly associated with CTP scan failure in AIS patients.

Genomic analyses of wild argali, domestic sheep, and their hybrids provide insights into chromosome evolution, phenotypic variation, and germplasm innovation.

Understanding the genetic mechanisms of phenotypic variation in hybrids between domestic animals and their wild relatives may aid germplasm innovation. Here, we report the high-quality genome assemblies of a male Pamir argali (O ammon polii, 2n = 56), a female Tibetan sheep (O aries, 2n = 54), and a male hybrid of Pamir argali and domestic sheep, and the high-throughput sequencing of 425 ovine animals, including the hybrids of argali and domestic sheep. We detected genomic synteny between Chromosome 2 of sheep and two acrocentric chromosomes of argali. We revealed consistent satellite repeats around the chromosome breakpoints, which could have resulted in chromosome fusion. We observed many more hybrids with karyotype 2n = 54 than with 2n = 55, which could be explained by the selfish centromeres, the possible decreased rate of normal/balanced sperm, and the increased incidence of early pregnancy loss in the aneuploid ewes or rams. We identified genes and variants associated with important morphological and production traits (e.g., body weight, cannon circumference, hip height, and tail length) that show significant variations. We revealed a strong selective signature at the mutation (c.334C > A, p.G112W) in TBXT and confirmed its association with tail length among sheep populations of wide geographic and genetic origins. We produced an intercross population of 110 F2 offspring with varied number of vertebrae and validated the causal mutation by whole-genome association analysis. We verified its function using CRISPR-Cas9 genome editing. Our results provide insights into chromosomal speciation and phenotypic evolution and a foundation of genetic variants for the breeding of sheep and other animals.

Comparative investigation of a lattice Boltzmann boundary treatment of multiphase mass transport with heterogeneous chemical reactions.

Multiphase reactive transport in porous media is an important component of many natural and engineering processes. In the present study, boundary schemes for the continuum species transport-lattice Boltzmann (CST-LB) mass transport model and the multicomponent pseudopotential model are proposed to simulate heterogeneous chemical reactions in a multiphase system. For the CST-LB model, a lattice-interface-tracking scheme for the heterogeneous chemical reaction boundary is provided. Meanwhile, a local-average virtual density boundary scheme for the multicomponent pseudopotential model is formulated based on the work of Li et al. [Li, Yu, and Luo, Phys. Rev. E 100, 053313 (2019)10.1103/PhysRevE.100.053313]. With these boundary treatments, a numerical implementation is put forward that couples the multiphase fluid flow, interfacial species transport, heterogeneous chemical reactions, and porous matrix structural evolution. A series of comparison benchmark cases are investigated to evaluate the numerical performance for different pseudopotential wetting boundary treatments, and an application case of multiphase dissolution in porous media is conducted to validate the present models' ability to solve complex problems. By applying the present LB models with reasonable boundary treatments, multiphase reactive transport in various natural or engineering scenarios can be simulated accurately.

Incremental value of Alberta Stroke Program Early CT Score to collateral score for predicting target mismatch in stroke patients with extended time window or unknown onset time.

PURPOSE: To evaluate whether Alberta Stroke Program Early CT Score (ASPECTS) could provide incremental value to collateral score, and their integration could be an effective surrogate of CTP in predicting target mismatch. MATERIAL AND METHODS: One hundred and fifty-nine stroke patients (onset time 6-16 h or with unknown onset time) with MCA and/or ICA occlusion underwent non-contrast computed tomography (NCCT) and CT perfusion (CTP) scan for initial assessment. Simulated single-phase CT angiography (sCTA, peak arterial phase) and multiphase CTA (mCTA) were reconstructed from CTP. ASPECTS was assessed on NCCT and sCTA. Collateral score was evaluated on mCTA. Target mismatch was defined as infarct core volume < 70 mL, the mismatch ratio ≥ 1.8, and the absolute mismatch volume ≥ 15 mL. Pearson correlation analysis, Mann-Whitney U test, chi-square test, and receiver operating characteristic curve analyses were performed. RESULTS: Median CTA source image (CTA-SI) ASPECTS was significantly lower than NCCT ASPECTS (p = 0.001). NCCT ASPECTS, CTA-SI ASPECTS, and mCTA collateral score correlated significantly with infarct core volume and mismatch ratio (all p < 0.05). Mismatch group showed significantly higher NCCT ASPECTS, CTA-SI ASPECTS, and mCTA collateral score than non-mismatch group (all p < 0.001). NCCT ASPECTS and CTA-SI ASPECTS showed comparable predicting performance with mCTA collateral score (p > 0.05). Adding CTA-SI ASPECTS to mCTA collateral score improved the performance of mCTA in predicting target mismatch (area under curve, 0.905 vs. 0.804, p = 0.003). CONCLUSION: ASPECTS can provide incremental information to collateral score in predicting target mismatch. If CTP scan fails, clinical decision based on ASPECTS and collateral score might be reasonable.

Cerebral blood volume Alberta Stroke Program Early Computed Tomography Score predicts intracranial hemorrhage after thrombectomy in patients with acute ischemic stroke in an extended time window.

BACKGROUND: Higher baseline Alberta Stroke Program Early Computed Tomography Score (ASPECTS) was associated with a lower probability of hemorrhagic transformation in patients with acute ischemic stroke (AIS). PURPOSE: To investigate the predictive value of cerebral blood volume (CBV)-ASPECTS of intracranial hemorrhage (ICH) in AIS treated with thrombectomy selected by computed tomographic perfusion (CTP) in an extended time window. MATERIAL AND METHODS: A total of 91 consecutive patients with AIS with large vessel occlusion in the anterior circulation after thrombectomy in an extended time window were enrolled between January 2018 and September 2019. ICH was diagnosed according to Heidelberg Bleeding Classification. CBV-ASPECTS was assessed by evaluating each ASPECTS region for relatively low CBV value compared with the mirror region in the contralateral hemisphere. Demographic characteristics, clinical data, CBV-ASPECTS, and procedure process and results were compared between patients with ICH and those without. RESULTS: ICH occurred in 31/91 (34.1%) patients with AIS. Symptomatic ICH (sICH) was observed in 4 (4.4%) patients, while asymptomatic ICH (aICH) was seen in 27 (29.7%). In univariate analysis, both ICH and aICH were associated with high admission NIHSS score (P<0.001 and P<0.001, respectively), more passes of retriever (P = 0.007 and P = 0.019, respectively), low NCCT-ASPECTS (P = 0.013 and P = 0.034, respectively), and low CBV-ASPECTS (P < 0.001 and P < 0.001, respectively). After multivariable analysis, low CBV-ASPECTS remained an independent predictor of ICH (odds ratio [OR] 0.521, 95% confidence interval [CI] 0.371-0.732, P < 0.001) and aICH (OR 0.532, 95% CI 0.376-0.752, P < 0.001), respectively. CONCLUSION: Low CBV-ASPECTS independently predicts ICH in patients with AIS treated with thrombectomy selected by CTP in an extended time window.

Lattice Boltzmann modeling of interfacial mass transfer in a multiphase system.

In the present study, a numerical model based on the lattice Boltzmann method (LBM) is proposed to simulate multiphase mass transfer, referred to as the CST-LB model. This model introduced continuum species transfer (CST) formulation by an additional collision term to model the mass transfer across the multiphase interface. The boundary condition treatment of this model is also discussed. In order to verify the applicability, the CST-LB model is combined with the pseudopotential multiphase model to simulate a series of benchmark cases, including concentration jump near the interface, gas dissolution in a closed system, species transport during drainage in a capillary tube, and multiphase species transport in the porous media. This CST-LB model can also be coupled with other multiphase LBMs since the model depends on the phase fraction field, which is not explicitly limited to specified multiphase models.

Asymptomatic Pulmonary Embolism After Shoulder Arthroscopy: Case Report and Literature Review.

BACKGROUND: Pulmonary embolism (PE) is extremely rare after shoulder arthroscopy. However, early identification of the situation deserves attention due to its potential risk of causing death. By now, it is still difficult to detect the PE without symptoms and clear sources during the perioperative period. CASE PRESENTATION: We report here two cases of asymptomatic PE, both happening within 24 h after shoulder arthroscopy, without any detected deep venous thrombosis of extremities. It is suspected the cases were due to the abnormal decrease in partial pressures of oxygen and arterial oxygen saturation, and were confirmed by computed tomography pulmonary angiography. We also discuss the reason why the patients showed no related symptoms when PE occurred and perform a review of PE following shoulder arthroscopy. CONCLUSIONS: This report highlights that PE could occur in the early phase after shoulder arthroscopy. An unexplained decrease in partial pressure of oxygen or arterial oxygen saturation should be considered seriously. The symptoms of PE might be masked due to patients' tolerance to hypoxia.

Macrophage-Derived Exosomes Promote Bone Mesenchymal Stem Cells Towards Osteoblastic Fate Through microRNA-21a-5p.

The effective healing of a bone defect is dependent on the careful coordination of inflammatory and bone-forming cells. In the current work, pro-inflammatory M1 and anti-inflammatory M2 macrophages were co-cultured with primary murine bone mesenchymal stem cells (BMSCs), in vitro, to establish the cross-talk among polarized macrophages and BMSCs, and as well as their effects on osteogenesis. Meanwhile, macrophages influence the osteogenesis of BMSCs through paracrine forms such as exosomes. We focused on whether exosomes of macrophages promote osteogenic differentiation. The results indicated that M1 and M2 polarized macrophage exosomes all can promote osteogenesis of BMSCs. Especially, M1 macrophage-derived exosomes promote osteogenesis of BMSCs through microRNA-21a-5p at the early stage of inflammation. This research helps to develop an understanding of the intricate interactions among BMSCs and macrophages, which can help to improve the process of bone healing as well as additional regenerative processes by local sustained release of exosomes.

Hyperperfusion on Arterial Spin Labeling MRI Predicts the 90-Day Functional Outcome After Mechanical Thrombectomy in Ischemic Stroke.

BACKGROUND: The prognostic significance of hyperperfusion after reperfusion therapy in patients with acute ischemic stroke (AIS) remains controversial. PURPOSE: To investigate the clinical factors associated with hyperperfusion, and the 90-day prognostic value of hyperperfusion after mechanical thrombectomy in AIS patients. STUDY TYPE: Retrospective. POPULATION/SUBJECTS: Fifty-four AIS patients who underwent mechanical thrombectomy. FIELD STRENGTH/SEQUENCE: Time-of-flight MR angiography, pulsed arterial spin labeling (ASL), diffusion-weighted imaging (DWI), and susceptibility-weighted imaging were performed at 3.0T within 1 week after thrombectomy. ASSESSMENT: Clinical factors including demographics, risk factors, stroke and treatment characteristics were collected and assessed. Hyperperfusion on ASL was defined as a focal increased cerebral blood flow on the affected side ≥130% of its mirror counterpart. Good clinical outcome at 90 days was defined as modified Rankin Scale score of 0-2. STATISTICAL TESTS: The interrater agreement was assessed using Cohen's kappa or the intraclass correlation coefficient. The relationship between hyperperfusion and clinical factors were analyzed by appropriate univariate statistics. Predictors of 90-day functional outcome were assessed by univariate analyses followed by multivariate logistic regression analysis and receiver-operating-characteristic curves. RESULTS: Thirty-six (66.7%) patients developed hyperperfusion on ASL after thrombectomy. Hyperperfusion was significantly correlated with successful recanalization (P < 0.05) and improvement of National Institutes of Health Stroke Scale scores at 24 hours (NIHSS24h ) (P < 0.05). A higher incidence of hemorrhage transformation was observed in patients with hyperperfusion than those without (63.9% vs. 50.0%), but no significant difference was found (P = 0.327). NIHSS24h (odds ratio [OR], 0.75, [95% confidence interval [CI] 0.62-0.91], P < 0.05), lesion volume on diffusion-weighted imaging (OR, 0.97, [95% CI 0.95-1.00], P < 0.05), and hyperperfusion on ASL (OR, 9.8, [95% CI 1.7-55.3], P < 0.05) were independent variables for predicting good functional outcomes. DATA CONCLUSION: Hyperperfusion on ASL correlated with successful recanalization and may be an independent prognostic marker for good neurological outcomes at 90 days in AIS patients after mechanical thrombectomy. LEVEL OF EVIDENCE: 4 TECHNICAL EFFICACY STAGE: 2.

Phytosphinganine Affects Plasmodesmata Permeability via Facilitating PDLP5-Stimulated Callose Accumulation in Arabidopsis.

Plant plasmodesmata (PDs) are specialized channels that enable communication between neighboring cells. The intercellular permeability of PDs, which affects plant development, defense, and responses to stimuli, must be tightly regulated. However, the lipid compositions of PD membrane and their impact on PD permeability remain elusive. Here, we report that the Arabidopsis sld1 sld2 double mutant, lacking sphingolipid long-chain base 8 desaturases 1 and 2, displayed decreased PD permeability due to a significant increase in callose accumulation. PD-located protein 5 (PDLP5) was significantly enriched in the leaf epidermal cells of sld1 sld2 and showed specific binding affinity to phytosphinganine (t18:0), suggesting that the enrichment of t18:0-based sphingolipids in sld1 sld2 PDs might facilitate the recruitment of PDLP5 proteins to PDs. The sld1 sld2 double mutant seedlings showed enhanced resistance to the fungal-wilt pathogen Verticillium dahlia and the bacterium Pseudomonas syringae pv. tomato DC3000, which could be fully rescued in sld1 sld2 pdlp5 triple mutant. Taken together, these results indicate that phytosphinganine might regulate PD functions and cell-to-cell communication by modifying the level of PDLP5 in PD membranes.

Effect of Huoxue Ointment on Osteoarthritis Caused by Knee Joint Injury in Rabbits / 中国实验方剂学杂志

Objective:To investigate the efficacy and anti-inflammatory and analgesic effect of Huoxue ointment on osteoarthritis of the knee caused by knee joint injury in rabbits. Method:A total of 60 big ear white rabbits were selected, with half male and half female. Ten rabbits were randomly selected by weight and set as normal control group. For the remaining rabbits, the injury of articular cartilage and anterior cruciate ligament were caused in the femoral ankle joint surface of the right hind knee joint, so as to establish an animal model of knee osteoarthritis in rabbits. For the normal control group, the skin was cut open and immediately sutured. Two weeks after the operation, rabbits were randomly divided into model control group according to body weight，positive voltaren group (1 cm·kg-1·d-1), and high-dose (6 g·kg-1·d-1), medium-dose (3 g·kg-1·d-1) and low-dose(1.5 g·kg-1·d-1) Huoxue ointment groups, with 10 animals in each group. Interleukinin-1β （IL-1β） in rabbit articular cavity fluid and peripheral blood were measured, contents of tumor necrosis factor-α （TNF-α） and free radical nitric oxide（NO）, rabbit articular cartilage was examined by histopathology, in order to study the efficacy of Huoxue ointment on rabbit knee arthritis. The model of increased permeability of capillaries in abdominal cavity of mice induced by acetic acid and the model of foot swelling of rats induced by carrageenan were used to determine the absorbance of mice peritoneal fluid and the rat foot swelling value, and study the anti-inflammatory effect of huoxue ointment. A total of 120 SPF ICR mice and 60 SD rats were selected, with half male and half female. The mice were randomly divided according to body weight into model control groups, positive Voltaren group (3 cm·kg-1·d-1), and high-dose (16 g·kg-1·d-1), medium-dose (8 g·kg-1·d-1), low-dose (4 g·kg-1·d-1) Huoxue ointment groups，with 12 animals in each group. The rats were randomly divided into the model control group, positive voltaren group (2 cm·kg-1·d-1), and high-dose (8 g·kg-1·d-1), medium-dose (4 g·kg-1·d-1), low-dose (2 g·kg-1·d-1) Huoxue ointment groups，with 10 animals in each group. Result:The tested dose of Huoxue ointment，the levels of IL-1β, TNF-α and NO in rabbit serum and joint fluid were decreased to varying degrees, and the effect was significant in the high-dose group (P<0.05). According to the pathological examination, the tested dose of Huoxue ointment, knee cartilage and synovial tissue lesions were significantly lower than those of the model control group (P<0.05)，the value of peritoneal fluid concentration and the foot swelling in medium and high-dose Huoxue ointment groups were significantly lower than those in the model control group (P<0.05). The analgesic effect of tail shaking in mice, and the pain threshold rate of the high-dose Huoxue ointment group were significantly higher than those of the model control group (P<0.05)， the analgesic effect lasted for 4 hours, the foot tenderness of rats in the large and medium-dose Huoxue ointment groups was significantly higher than that in the model control group (P<0.05). Conclusion:Huoxue ointment has obvious joint repair and anti-inflammatory and analgesic effects on osteoarthritis of knee caused by knee joint injury in rabbits.

Effect and Mechanism of Four Kinds of Traditional Chinese Medicine Moisturizers Based on Shaoyao Gancaotang on Atopic Dermatitis / 中国实验方剂学杂志

Objective:To investigate the therapeutic effect and possible mechanism of four types of Chinese herbal moisturizers made in laboratory for atopic dermatitis induced by 2，4-dinitrofluorobenzene （DNFB） in mice. Method:According to the body weight， BALA/c mice were randomly divided into normal group， model group， blank cream group （moisturizer A）， Shaoyao Gancaotang group （moisturizer B）， Shaoyao Gancaotang with Portulacae Herba，Ginseng Radix et Rhizoma and Honeysuckle Stem group （moisturizer C）， and Shaoyao Gancaotang with Ginseng Radix et Rhizoma and Honeysuckle Stem group （moisturizer D） ， with the dose of 25 g·kg-1 per day， as well as tacrolimus ointment group of 3 g·kg-1 per day， with 10 to 12 mice in each group. Except the normal group， the mice in the other groups were treated with 0.5% DNFB in the hair removal skin of back， 100 μL each for 7 days. Starting from the 7th day， each group was given the appropriate skin cream for external use intervention， once per day， for 15 consecutive days， except for the normal and the model groups. The animal body mass was measured once a week， and the animal back skin was graded three times a week， and the skin lesion score was recorded. After the mice were killed， the left and right ears were taken， the weight of both ears was punched and the degree of swelling was calculated. The back skin was fixed and stained with hematoxylin-eosin（HE） method， and then pathologic examination was conducted to observe and score the pathological changes of mouse back skin. Blood was obtained after the last dose and enzyme-linked immunosorbent assay （ELISA） was used to determine the immunoglobulin（Ig）E content in serum. Western blot was used to measure the expression of signal transduction and activator of transcription 3 （STAT3）， phosphorylation （p）-STAT3 in the skin tissue. Result:Compared with the normal group， the body mass decreased continuously， a series of inflammatory changes such as erythema， edema， dryness， desquamation and callus exfoliation and so on occurred in the modeling area， and the skin lesion score increased significantly in the model group. Additionally， the cuticle of ear skin was thickened and the degree of ear swelling was obviously increased in the model group. Microscopically， the occurred changes in the model mice included the local necrosis of the epidermis， epidermal thickening， epidermal hyperplasia， and the hyperkeratosis and hypokeratosis in the cuticle， as well as the subcutaneous inflammatory cell infiltration and so on. Furthermore， the content of serum IgE andthe expression of p-STAT3 in skin tissues increased significantly in the model group. Compared with the model group， the body mass of mice in group C and D was significantly increased （P<0.01）， and the skin lesion status score was decreased （P<0.05，P<0.01）.The degree of auricle swelling was significantly reduced in group B， C and D compared with that in the model group （P<0.01）.The degree of skin necrosis and defect and epidermal hyperplasia of mice in moisturizer C group was significantly reduced compared with that in model group （P<0.05，P<0.01）. Serum IgE levels of mice in group C and D were significantly lower than those in the model group （P<0.05，P<0.01）. The expression of p-STAT3 protein in skin tissues of mice in moisturizer C group was significantly lower than that in model group （P<0.05）. Conclusion:The moisturizers B， C and D all have certain therapeutic effect on atopic dermatitis， among which moisturizers C has the most obvious therapeutic effect. The possible mechanism may be that it reduces the level of inflammatory cytokines by inhibiting the increase of serum IgE content and the phosphorylation of STAT3.

[Interaction between necroptosis and apoptosis in MC3T3-E1 cell death induced by dexamethasone].

OBJECTIVE: To investigate the relationship between necroptosis and apoptosis in MCET3-E1 cell death induced by glucocorticoids. METHODS: MC3T3-E1 cells were incubated with 10-6 mol/L dexamethasone followed by treatment with the apoptosis inhibitor z-VAD-fmk (40 µmol/L) or the necroptosis inhibitor necrostatin-1 (40 µmol/L) for 2 h. At 72 h after incubation with dexamethasone, the cells were harvested to determine the cell viability using WST-1 assay and the rate of necrotic cells using annexin V/PI double staining; the percentage of apoptotic cells was determined using Hoechst staining. The mitochondrial membrane potential and the level of ATP in the cells were also evaluated. Transmission electron microscopy was used to observe the microstructural changes of the cells. The expressions of RIP-1 and RIP-3 in the cells were detected by Western blotting. RESULTS: At a concentration of 10-6 mol/L, dexamethasone induced both apoptosis and necroptosis in MC3T3- E1 cells. Annexin V/PI double staining showed that inhibition of cell apoptosis caused an increase in cell necrosis manifested by such changes as mitochondrial swelling and plasma membrane disruption, as shown by electron microscopy; Hoechst staining showed that the percentage of apoptotic cells was significantly reduced. When necroptosis was inhibited by necrostatin-1, MC3T3-E1 cells showed significantly increased apoptosis as shown by both AV/PI and Hoechst staining, and such changes were accompanied by changes in mitochondrial membrane potential and ATP level in the cells. CONCLUSIONS: In the process of dexamethasone-induced cell death, necroptosis and apoptosis can transform reciprocally accompanied by functional changes of the mitochondria.