Inhibition of HMGB1 suppresses inflammation and catabolism in temporomandibular joint osteoarthritis via NF-κB signaling pathway.

HMGB1 is a highly conserved nuclear protein that is rapidly released into the extracellular environment during infection or tissue damage. In osteoarthritis, HMGB1 acts as a pro-inflammatory cytokine inducing a positive feedback loop for synovial inflammation and cartilage degradation. The aim of this study was to explore the role of HMGB1 in inflammation and catabolism of temporomandibular joint osteoarthritis (TMJOA) and whether inhibition of HMGB1 affects TMJOA. Human synovial fibroblasts were incubated with HMGB1, the expression of pro-inflammatory cytokines and catabolic mediators were measured by Western blot and ELISA. NF-κB signaling pathway involvement was studied by the NF-κB inhibitor and detected by Western blotting and immunofluorescence staining. TMJOA was induced by an injection of Complete Freund's adjuvant (CFA) into anterosuperior compartment of rat's joint. An anti-HMGB1 antibody was used to assess the effect to HMGB1 in the synovium and cartilage of the CFA-induced TMJOA rats by H&E, Safranin O, Masson trichrome staining, immunohistochemistry and immunofluorescence. HMGB1 markedly increased the production of MMP13, ADAMTS5, IL-1ß and IL-6 through activating NF-κB signaling pathway in human synovial fibroblasts. In vivo, application of the HMGB1 neutralizing antibody effectively ameliorated the detrimental extent of TMJOA. Furthermore, the HMGB1 neutralizing antibody reduced the expression of NF-κB, pro-inflammatory cytokines and catabolic mediators in the synovium and cartilage of CFA-induced TMJOA rats. HMGB1 inhibition alleviates TMJOA by reducing synovial inflammation and cartilage catabolism possibly through suppressing the NF-κB signaling pathway and may become a therapeutic method against TMJOA.

Diverse Triterpenoids from Mastic Produced by Pistacia lentiscus and Their Anti-Inflammatory Activities.

Eight undescribed triterpenoids (1-8), including one apotirucallane-type triterpenoid (1), six tirucallane-type triterpenoids (2-7), and one oleanane-type triterpenoid (8), along with ten known compounds (9-18) were isolated from the resins of Pistacia lentiscus. Their structures were elucidated by integrating NMR spectroscopic analyses and ESI-HR-MS. Compounds 5, 11-17 exhibited moderate inhibitory abilities against NO production in LPS-induced RAW264.7 cells, with IC50 values in the range of 18.26-50.37 µM, compared to that of the positive control dexamethasone (IC50 =20.24 µM).

Euphatexols C - G, five new triterpenoids from the latex of Euphorbia resinifera.

Five new triterpenoids named as euphatexols C - G (1-5) were isolated from the latex of Euphorbia resinifera. Their chemical structures were determined by extensive spectroscopic analysis, including IR, HRMS, 1 D and 2 D NMR data. Compounds 1-5 showed moderate anti-inflammatory activity with IC50 values of 22.30, 48.04, 21.89, 38.15 and 41.15 µM, respectively.

[Seasonal Variation and Source Apportionment of Carbonaceous Species in PM2.5 in Chengde].

In order to study the seasonal variations and pollution sources of carbonaceous species in PM2.5 in Chengde, the concentration of these components was determined in atmospheric PM2.5 samples collected in January, April, July, and October 2019. The change in carbonaceous species were analyzed based on the estimation of the ratio of organic carbon(OC) to elemental carbon(EC), total carbonaceous aerosol(TCA), and secondary organic carbon(SOC). The source of these pollutants was determined by means of the backward trajectory and principal component analysis(PCA). The results showed that the mean mass concentrations of PM2.5, OC, and EC during the sampling period were(31.26±21.39) µg·m-3,(13.27±8.68) µg·m-3, and(2.80±1.95) µg·m-3, respectively. The seasonal variations of PM2.5 were:winter[(47.68±30.37) µg·m-3]>autumn[(28.72±17.12) µg·m-3]>spring[(26.59±15.32) µg·m-3]>summer[(23.17±8.38) µg·m-3], consistent with the trend of total carbon(TC), OC, and EC. The source of OC and EC during winter(R2=0.85) was similar. Based on the ratio of OC/EC, all four seasons were affected by traffic and coal-burning source emissions, and the most affected season by bituminous coal emissions was winter. The average concentration of TCA was(21.38±13.68) µg·m-3, which accounted for 68.39% of PM2.5. The order of secondary conversion rate(SOC/OC) was:spring(54.09%) >autumn(37.64%) >summer(32.91%) >winter(25.43%). The results of the backward trajectory simulation show that the pollutant concentrations carried by air masses are relatively low in spring and summer, and the transport channels of pollutants are southwest in autumn and northwest in winter. The results of the PCA showed that the key to reducing PM2.5 in Chengde is to control emissions from vehicle exhausts, and coal and biomass combustion sources.

[Four new lanostane triterpenoids from latex of Euphorbia resinifera].

Four new lanostane triterpenoids, 3ß-hydroxy-12α-methoxylanosta-7,9(11),24-triene(1), 3ß-hydroxy-12α-methoxy-24-methylene-lanost-7,9(11)-dien(2), 3,7-dioxo-lanosta-8,24-diene(3), and 3,7-dioxo-24-methylene-lanost-8-en(4), were isolated from the latex of Euphorbia resinifera with a variety of chromatography methods. Their structures were elucidated based on spectroscopic data and/or comparison with the data reported in previous research. Compounds 1, 2, and 4 showed moderate inhibition of LPS-induced NO production by RAW264.7, with IC_(50) of 30.4, 37.5, and 28.3 µmol·L~(-1), respectively.

[Spatio-Temporal Evolution Characteristics and Source Apportionment of O3 and NO2 in Shijiazhuang].

Ground-level O3, NO2, and meteorological (temperature, humidity, wind speed, precipitation, and sunshine duration) data from 18 counties in Shijiazhuang City from 2014 to 2017, and volatile organic compounds (VOCs) data for Summer 2017, were analyzed to explore the spatial patterns, evolution, influencing factors, and source apportionment of O3 and NO2 in Shijiazhuang City. Network analysis and inverse distance weighted (IDW) spatial autocorrelation and backward trajectories analyses were performed. The results indicate that O3 concentrations increased between 2014 and 2017, and monthly variations showed a unimodal trend. The typical period of peak O3 pollution (O3 ≥ 160 µg·m-3) was from May to September, characterized by high temperatures, low humidity, weak winds, and strong solar radiation. The O3 concentrations were negatively correlated with the NO2. Furthermore, O3 concentrations increased year-on-year since 2015 in main urban area, and the dominant pollutant type had changed from NO2 (2014 to 2016) to VOCs (2016 and 2017). However, the O3 concentration of county-areas limited by the VOCs. The main factors affecting O3 concentrations were industry, agriculture, economy, and population, and centers of O3 pollution associated with secondary industry appeared in the main urban areas of Shijiazhuang and Luancheng. Moreover, VOCs trajectories during the summer monitoring period were clustered in three source directions:(A) East-northeast, 26.67%; (B) Northwest-west, 43.33%; and (C) Southeast-south, 30%). Trajectories (A) and (C) were the dominant directions of VOC transmission (east-southeast).

[Characteristics and Source Apportionment of VOCs and O3 in Shijiazhuang].

To study the composition characteristics and sources of volatile organic compounds (VOCs) in Shijiazhuang City, three national control points were selected to conduct VOCs sampling and analysis from March 2017 to January 2018. The correlation of VOCs through combination with meteorological and ground-level O3 data, and the sources of VOCs were analyzed by positive matrix factorization (PMF). To quantify the pollution period of O3 in summer, its temporal sequence characteristics were studied by wavelet analysis. During the sampling period, the average concentration of ambient total VOCs (TVOCs) was (137.23±64.62) µg·m-3. Haloalkanes were the most dominant VOC compounds, accounting for 31.77% of total VOCs mass, followed by aromatic (30.97%) and oxygenated VOCs (OVOCs, 23.76%). The seasonal variation in VOC concentration followed the trend in winter (187.7 µg·m-3) > autumn (146.8 µg·m-3) > spring (133.24 µg·m-3) > summer (107.1 µg·m-3); the concentration of VOCs shows a trend of increasing gradient from west to east. The O3 concentration correlated negatively with VOCs and NO2, and positively with temperature, sunshine duration, wind speed, and visibility. Changes in meteorological elements were concerned before the occurrence of ozone pollution in summer, especially in 4-5 days in June and 7-8 days during July to August after the occurrence of increasing temperature. Finally six potential sources of VOCs were quantified by the PMF model, including from gasoline emissions (24.78%), diesel vehicle emissions (24.69%), solvent usage (18.64%), the chemical industry (11.87%), regional background (10.84%), and the pharmaceutical industry (9.17%). Ozone formation potential (OFP) contribution of emission sources of gasoline and diesel vehicles (54.98%) was over half of the total contribution. Meanwhile, these findings illustrated that control of vehicle emissions and industrial sources would be an important way to reduce VOCs concentrations and improve air quality in Shijiazhuang.

Effect of silver diammine fluoride on micro-ecology of plaque from extensive caries of deciduous teeth - in vitro study.

BACKGROUND: The mechanism of action of silver diammine fluoride (SDF) on plaque micro-ecology is seldom studied. This study investigated micro-ecological changes in dental plaque on extensive caries of deciduous teeth after topical SDF treatment. METHODS: Deciduous teeth with extensive caries freshly removed from school children were collected in clinic. Unstimulated saliva collection and initial plaque sampling were done before tooth extraction, then each caries was topically treated with 38% SDF in vitro. After intervention, each tooth was stored respectively in artificial saliva at 37 °C. Repeated plaque collections were done at 24 h and 1 week post-intervention. Post-intervention micro-ecological changes including microbial diversity, microbial metabolism function as well as species correlations were analyzed and compared after pyrosequencing of the DNA from the plaque sample using Illumina MiSeq platform. RESULTS: After SDF application, microbial diversity decreased (P > 0.05), although not statistically significant. Microbial community composition post-intervention was noticeably different from that of supragingival and pre-intervention plaque as well as saliva. At 1 week post-intervention, the relative content of Pseudomonas, Fusobacterium and Pseudoramibacter were higher than before, while most of the other bacteria were reduced, although the changes were not statistically significant (P > 0.05). The inter-microbial associations became more complex, much more positive associations among survived bacteria were observed than negative ones. COG function classification diagram showed carbohydrate transportation and metabolic functions in the plaque were significantly reduced at 24 h and 1 week post-intervention. CONCLUSIONS: SDF has extensive antimicrobial effect on dental plaque, which may reduce carbohydrate metabolism in dental plaque and help promote new balance of the plaque flora.

The protective role of NR4A3 in acute myocardial infarction by suppressing inflammatory responses via JAK2-STAT3/NF-κB pathway.

Inflammatory responses play a critical role in left ventricular remodeling after acute myocardial infarction (AMI). NR4A3, a member of the NR4A orphan nucleus receptor family, has recently emerged as a therapeutic target for treatment of inflammation. This aim of this study is to explore the therapeutic effect of NR4A3 in cardiac remodeling post AMI. Male C57BL/6 mice were administered with lentiviral over-expression of NR4A3 (lenti-NR4A3) or empty vector (lenti-con) 7 days before coronary artery ligation. H9c2 cardiomyocytes deprived of serum were used to mimic ischemic conditions in vivo. Lenti-NR4A3 treatment significantly repressed neutrophil infiltration in the myocardium, reduced infarct size, and attenuated the reduction of left ventricular function after AMI. Furthermore, NR4A3 over-expression inhibited the NF-κB (IκB) signaling by decreasing IκBα phosphorylation and by inhibiting the translocation of p65 to the nucleus. Meanwhile, NR4A3 over-expression also increases the activity of JAK2-STAT3 signaling in mouse hearts after AMI. The inhibitory effect of NR4A3 on NF-κB activation was almost completely abolished by the JAK2 inhibitor AG490, indicating that NR4A3 prevented serum deprivation induced NF-κB activation in a STAT3 dependent manner. These findings provide novel evidence that NR4A3 could inhibit post-AMI inflammation responses via JAK2-STAT3/NF-κB signaling and may well be a therapeutic target for cardiac remodeling after AMI.

Antibody incubation at 37°C improves fluorescent immunolabeling in free-floating thick tissue sections.

Fluorescent immunolabeling and imaging in free-floating thick (50-60 µm) tissue sections is relatively simple in practice and enables design-based non-biased stereology, or 3-D reconstruction and analysis. This method is widely used for 3-D in situ quantitative biology in many areas of biological research. However, the labeling quality and efficiency of standard protocols for fluorescent immunolabeling of these tissue sections are not always satisfactory. Here, we systematically evaluate the effects of raising the conventional antibody incubation temperatures (4°C or 21°C) to mammalian body temperature (37°C) in these protocols. Our modification significantly enhances the quality (labeling sensitivity, specificity, and homogeneity) and efficiency (antibody concentration and antibody incubation duration) of fluorescent immunolabeling of free-floating thick tissue sections.

Behavioral characterization of neuropathic pain on the glabrous skin areas reinnervated solely by axotomy-regenerative axons after adult rat sciatic nerve crush.

In cranial and spinal nerve ganglia, both axotomized primary sensory neurons without regeneration (axotomy-nonregenerative neurons) and spared intact primary sensory neurons adjacent to axotomized neurons (axotomy-spared neurons) have been definitely shown to participate in pain transmission in peripheral neuropathic pain states. However, whether axotomized primary sensory neurons with regeneration (axotomy-regenerative neurons) would be integral components of neural circuits underlying peripheral neuropathic pain states remains controversial. In the present study, we utilized an adult rat sciatic nerve crush model to systematically analyze pain behaviors on the glabrous plantar surface of the hindpaw sural nerve skin territories. To the best of our knowledge, our results for the first time showed that heat hyperalgesia, cold allodynia, mechanical allodynia, and mechanical hyperalgesia emerged and persisted on the glabrous sural nerve skin areas after adult rat sciatic nerve crush. Interestingly, mechanical hyperalgesia was sexually dimorphic. Moreover, with our optimized immunofluorescence staining protocol of free-floating thick skin sections for wide-field epifluorescence microscopic imaging, changes in purely regenerative reinnervation on the same skin areas by axotomized primary sensory afferents were shown to be paralleled by those pathological pain behaviors. To our surprise, Protein Gene Product 9.5-immunoreactive nerve fibers with regular and large varicosities ectopically emigrated into the upper dermis of the glabrous sural nerve skin territories after adult rat sciatic nerve crush. Our results indicated that axotomy-regenerative primary sensory neurons could be critical elements in neural circuits underlying peripheral neuropathic pain states. Besides, our results implied that peripheral neuropathic pain transmitted by axotomy-regenerative primary sensory neurons alone might be a new dimension in the clinical therapy of peripheral nerve trauma beyond regeneration.

[Early Systemic Administration of IL-10 Inhibits Neuropathic Pain in Adult Rats with Tibial Nerve Injury].

OBJECTIVES: To determine the effect of early systemic administration of IL-10 on peripheral neuropathic pain induced by tibial nerve permanent transection [modified spared nerve injury (mSNI)]in adult rats. METHODS: Male adult Sprague-Dawley (SD) rats (ten-week old, 250-300 g) with mSNI were randomly divided into mSNI, sham-operated, IL-10 intervention (intraperitoneal injection), PBS intervention (intraperitoneal injection) groups, each containing six rats. Intraperitoneally injections (IL-10 or PBS) were given immediately after surgeries for a single regime with a dosage of 500 uL (0.1 mg/mL). Plantar test, von Frey hairs test, pinprick test and acetone test were performed before and after tibial nerve injuries (0 d, 4/5 d, 7/8 d, 14/15 d) to evaluate region-specific pain responses of the rats on the plantar sural and saphenous skin territories of ipsilateral and contralateral hindpaws. The hindpaw position (on 8 d) of six additional rats with standard SNI was compared with those with mSNI. RESULTS: The rats with standard SNI showed an eversion posture of hindpaws, more prominent than those with mSNI. Region-specific pathological pain evoked by mechanical and thermal stimuli on the sural and saphenous skin territories of the plantar surfaces of rat hindpaws was demonstrated on the ipsilateral rather than contralateral hindpaws. This effect was shown in the rats with mSNI but not in those with sham operations. Compared with PBS, early intraperitoneal injection of IL-10 significantly and persistently attenuated either allodynia or hyperalgesia in the rats with mSNI. CONCLUSIONS: Tibial nerve permanent transection models of adult rats can be used as a simple but useful rodent model of peripheral neuropathic pain. Early systemic administration of IL-10 impairs the pathogenesis of neuropathic pain induced by tibial nerve injuries.

[A prospective randomized control clinical trial about clopidogrel combined with warfarin versus clopidogrel alone in the prevention of restenosis after femoral-popliteal artery angioplasty].

OBJECTIVE: Using two antithrombotic treatment (clopidogrel vs. clopidogrel combined warfarin) strategies after femoral-popliteal artery angioplasty prospectively, to evaluate which strategy is more effective for the restenosis prevention. METHODS: Totally 50 patients referred for endovascular treatment (including the percutaneous transluminal angioplasty (PTA) and stent implantation) of the superficial femoral artery and popliteal artery from January 2008 to May 2009 were randomly divided into clopidogrel group (group A, 25 cases, 30 limbs) and clopidogrel plus warfarin group (group B, 25 cases, 33 limbs) before operation. Clinical outcomes and restenosis rate of the target lesions were evaluated at 3, 6 and 12 months after operation. RESULTS: Totally 88 patients were screened for participation in the study, 56 patients were included after the follow-up of 12 months. At 3 months, the rates of restenosis were 16.7% in group A and 18.2% in group B (χ² = 0.025, P = 0.874). At 6 months, the accumulated restenosis rates were 36.7% in group A and 36.4% in group B (χ² = 0.001, P = 0.98). At 12 months, the accumulated restenosis rates were 53.3% in group A and 42.4% in group B (χ² = 0.75, P = 0.387). Analysis for the critical limb ischemia sub-group showed that follow-up of 12 months, the accumulated restenosis rate was 8/10 in group A and 6/12 in group B (χ² = 1.023, P = 0.312). CONCLUSION: The clopidogrel alone treatment for PTA or PTA plus stent implantation of femoral popliteal artery has no statistically significant difference in comparison with the clopidogrel combined warfarin treatment in terms of the cumulative vascular restenosis rate at 3, 6, 12 months postoperatively.

A novel first aid stretcher for immobilization and transportation of spine injured patients.

Effective immobilization and transportation are vital to the life-saving acute medical care needed when treating critically injured people. However, the most common types of stretchers used today are wrought with problems that can lead to further medical complications, difficulty in employment and rescue, and ineffective transitions to hospital treatment. Here we report a novel first aid stretcher called the "emergency carpet", which solves these problems with a unique design for spine injured patients. Polyurethane composite material, obtained by a novel process of manually mixing isocyanate and additives, can be poured into a specially designed fabric bag and allowed to harden to form a rigid human-shaped stretcher. The effectiveness of the emergency carpet was examined in the pre-hospital management of victims with spinal fractures. Additionally, it was tested on flat ground and complex terrain as well as in the sea and air. We demonstrated that the emergency carpet can be assembled and solidified on the scene in 5 minutes, providing effective immobilization to the entire injured body. With the protection of the emergency carpet, none of the 20 patients, who were finally confirmed to have spinal column fracture or dislocation, had any neurological deterioration during transportation. Furthermore, the carpet can be handled and transported by multiple means under differing conditions, without compromising immobilization. Finally, the emergency carpet allows the critically injured patient to receive multiple examinations such as X-ray, CT, and MRI without being removed from the carpet. Our results demonstrate that the emergency carpet has ideal capabilities for immobilization, extrication, and transportation of the spine injured patients. Compared with other stretchers, it allows for better mobility, effective immobilization, remarkable conformity to the body, and various means for transportation. The emergency carpet is promising for its intrinsic advantages in the pre-hospital management of accident victims.

Glycomimetic improves recovery after femoral injury in a non-human primate.

In adult mammals, restoration of function after peripheral nerve injury is often poor and effective therapies are not available. Previously we have shown in mice that a peptide which functionally mimics the human natural killer cell (HNK)-1 trisaccharide epitope significantly improves the outcome of femoral nerve injury. Here we evaluated the translational potential of this treatment using primates. We applied a linear HNK-1 mimetic or a functionally inactive control peptide in silicone cuffs used to reconstruct the cut femoral nerves of adult cynomolgus monkeys (Macaca fascicularis). Functional recovery was evaluated using video-based gait analysis over a 160-day observation period. The final outcome was further assessed using force measurements, H-reflex recordings, nerve histology, and ELISA to assess immunoreactivity to HNK-1 in the treated monkeys. Gait deficits were significantly reduced in HNK-1 mimetic-treated compared with control peptide-treated animals between 60 and 160 days after injury. Better outcome at 160 days after surgery in treated versus control animals was also confirmed by improved quadriceps muscle force, enhanced H-reflex amplitude, decreased H-reflex latency, and larger diameters of regenerated axons. No adverse reactions to the mimetic, in particular immune responses resulting in antibodies against the HNK-1 mimetic or immune cell infiltration into the damaged nerve, were observed. These results indicate the potential of the HNK-1 mimetic as an efficient, feasible, and safe adjunct treatment for nerve injuries requiring surgical repair in clinical settings.

Early neurosurgical intervention of spinal cord contusion: an analysis of 30 cases.

BACKGROUND: The incidence of spinal injury with spinal cord contusion is high in developed countries and is now growing in China. Furthermore, spinal cord injury happens mostly in young people who have a long life expectance. A large number of patients thus are wheelchair bound for the rest of their lives. Therefore, spinal cord injury has aroused great concern worldwide. Despite great efforts, recovery from spinal cord injury remains unsatisfactory. Based on the pathology of spinal cord contusion, an idea of early neurosurgical intervention has been formulated in this study. METHODS: A total of 30 patients with "complete" spinal cord injury or classified as American Spinal Injury Association (ASIA)-A were studied. Orthopedic treatment of the injured vertebra (e), internal fixation of the vertebral column, and bilateral laminectomy for epidural decompression were followed directly by neurosurgical management, including separation of the arachnoid adhesion to restore cerebrospinal fluid flow and debridement of the spinal cord necrotic tissue with concomitant intramedullary decompression. Rehabilitation started 17 days after the operation. The final outcome was evaluated after 3 months of rehabilitation. Pearson chi-square analysis was used for statistical analysis. RESULTS: All the patients recovered some ability to walk. The least recovered patients were able to walk with a wheeled weight support and help in stabilizing the weight bearing knee joint (12 cases, 40%). Thirteen patients (43%) were able to walk with a pair of crutches, a stick or without any support. The timing of the operation after injury was important. An optimal operation time window was identified at 4 - 14 days after injury. CONCLUSIONS: Early neurosurgical intervention of spinal cord contusion followed by rehabilitation can significantly improve the locomotion of the patients. It is a new idea of a therapeutic approach for spinal cord contusion and has been proven to be very successful.

[Synthesis and characteristic of cobalt bearing ferrite particles at room temperature].

Cobalt bearing magnetite particles were synthesized by aerial oxidation of ferrous and cobalt ions at alkaline condition and room temperature. The samples obtained at different time intervals during reaction were subjected to XRD and IR, and it was indicated that the cobalt-bearing ferrites were produced through green-rust intermediate phase. Different compounds were synthesized using 1/20, 1/15, 1/10, 1/7, 1/5 and 1/3 Co2+/Fe2+ starting ratios, and Co-ferrites with Co/Fe of 0.05, 0.06, 0.08, 0.12, 0.17, 0.18 were obtained. The chemical composition, microstructure and micrograph of those precipitates were characterized by chemical analysis, IR, XRD and SEM. The results revealed that Co2+ replaced Fe3+ at octahedral position in spinel structure, and the increase of Co in initial solution showed deleterious effect on the crystallinity of the precipitates.