Matrix-transmitted paratensile signaling enables myofibroblast-fibroblast cross talk in fibrosis expansion.

While the concept of intercellular mechanical communication has been revealed, the mechanistic insights have been poorly evidenced in the context of myofibroblast-fibroblast interaction during fibrosis expansion. Here we report and systematically investigate the mechanical force-mediated myofibroblast-fibroblast cross talk via the fibrous matrix, which we termed paratensile signaling. Paratensile signaling enables instantaneous and long-range mechanotransduction via collagen fibers (less than 1 s over 70 µm) to activate a single fibroblast, which is intracellularly mediated by DDR2 and integrin signaling pathways in a calcium-dependent manner through the mechanosensitive Piezo1 ion channel. By correlating in vitro fibroblast foci growth models with mathematical modeling, we demonstrate that the single-cell-level spatiotemporal feature of paratensile signaling can be applied to elucidate the tissue-level fibrosis expansion and that blocking paratensile signaling can effectively attenuate the fibroblast to myofibroblast transition at the border of fibrotic and normal tissue. Our comprehensive investigation of paratensile signaling in fibrosis expansion broadens the understanding of cellular dynamics during fibrogenesis and inspires antifibrotic intervention strategies targeting paratensile signaling.

Cryotherapy on Subjective Sleep Quality, Muscle, and Inflammatory Response in Chinese Middle- and Long-Distance Runners After Muscle Damage.

ABSTRACT: Qu, C, Wu, Z, Xu, M, Lorenzo, S, Dong, Y, Wang, Z, Qin, F, and Zhao, J. Cryotherapy on subjective sleep quality, muscle, and inflammatory response in Chinese middle- and long-distance runners after muscle damage. J Strength Cond Res 36(10): 2883-2890, 2022-The purpose of this investigation was to explore the effects of cold-water immersion (CWI), contrast-water therapy (CWT), and whole-body cryotherapy (CRY) on subjective sleep quality, muscle damage markers, and inflammatory markers in middle- and long-distance runners after muscle damage. Twelve male runners from Beijing Sport University completed a muscle damage exercise protocol and were treated with different recovery methods (CWI, CWT, CRY, or control [CON]) immediately after exercise and at 24-, 48-, and 72-h postexercise. The Pittsburgh Sleep Quality Index questionnaire score, lactate dehydrogenase (LDH) activity, myoglobin (Mb) activity, interleukin-6 (IL-6) activity, and soluble intercellular adhesion molecule-1 (sICAM-1) activity were measured at 7 time points (preexercise; immediately postexercise; and at 1-, 24-, 48-, 72-, and 96-h postexercise). Pittsburgh Sleep Quality Index scores indicated that the CRY condition had improved sleep quality compared with the CON and CWI conditions (p < 0.05). In terms of LDH activity, the CRY and CWT conditions had improved recovery compared with the CON and CWI conditions (p < 0.05). In terms of Mb activity, the CRY condition exhibited improved recovery compared with that of the CON and CWI conditions (p < 0.05), and the CWT condition showed better recovery than that of the CON condition (p < 0.05). In terms of IL-6 activity, the CRY condition showed improved recovery compared with the CWI condition (p < 0.05). Finally, in terms of sICAM-1 activity, the CRY condition had enhanced recovery compared with the other 3 conditions (p < 0.05). The results from this study suggest that CRY improves subjective sleep quality and reduces muscle damage and inflammatory responses in middle- and long-distance runners. In addition, CWT reduced muscle damage and inflammatory responses, but its effects on the other parameters were inconclusive.

A Mixed-Method Approach of Pre-Cooling Enhances High-Intensity Running Performance in the Heat.

We investigated whether single or combined methods of pre-cooling could affect high-intensity exercise performance in a hot environment. Seven male athletes were subjected to four experimental conditions for 30 min in a randomised order. The four experimental conditions were: 1) wearing a vest cooled to a temperature of 4 ℃ (Vest), 2) consuming a beverage cooled to a temperature of 4 ℃ (Beverage), 3) simultaneous usage of vest and consumption of beverage (Mix), and 4) the control trial without pre-cooling (CON). Following those experimental conditions, they exercised at a speed of 80% VO2max until exhaustion in the heat (38.1 ± 0.6 ℃, 55.3 ± 0.3% RH). Heart rate (HR), rectal temperature (Tcore), skin temperature (Tskin), sweat loss (SL), urine specific gravity (USG), levels of sodium (Na+) and potassium (K+), rating of perceived exertion (RPE), thermal sensation (TS), and levels of blood lactic acid ([Bla]) were monitored. Performance was improved using the mixed pre-cooling strategy (648.43 ± 77.53 s, p = 0.016) compared to CON (509.14 ± 54.57 s). Tcore after pre-cooling was not different (Mix: 37.01 ± 0.27 ℃, Vest: 37.19 ± 0.33 ℃, Beverage: 37.03 ± 0.35 ℃) in all cooling conditions compared to those of CON (37.31 ±0.29 ℃). A similar Tcore values was achieved at exhaustion in all trials (from 38.10 ℃ to 39.00 ℃). No difference in the level of USG was observed between the conditions. Our findings suggest that pre-cooling with a combination of cold vest usage and cold fluid intake can improve performance in the heat.

[Environment of high temperature or air particle matter pollution, and health promotion of exercise].

It is important to keep human health in special environment, since the special environment has different effects on health. In this review, we focused on high temperature and air particle matter environment, and health promotion of exercise. Exercise and high temperature are the main non-pharmacological therapeutic interventions of insulin resistance (IR). PGC-1α is key regulatory factor in health promotion of exercise and high temperature. The novel hormone Irisin might be the important pathway through which heat and exercise could have positive function on IR. Air particle matter (PM) is associated with onset of many respiratory diseases and negative effects of exerciser performance. However, regular exercise plays an important role in improving health of respiratory system and lowering the risk induced by PM. Furthermore, free radicals and inflammatory pathways are included in the possible mechanisms of positive physiological effects induced by exercise in air particle matter environment.

Expansion of human megakaryocyte-biased hematopoietic stem cells by biomimetic Microniche.

Limited numbers of available hematopoietic stem cells (HSCs) limit the widespread use of HSC-based therapies. Expansion systems for functional heterogenous HSCs remain to be optimized. Here, we present a convenient strategy for human HSC expansion based on a biomimetic Microniche. After demonstrating the expansion of HSC from different sources, we find that our Microniche-based system expands the therapeutically attractive megakaryocyte-biased HSC. We demonstrate scalable HSC expansion by applying this strategy in a stirred bioreactor. Moreover, we identify that the functional human megakaryocyte-biased HSCs are enriched in the CD34+CD38-CD45RA-CD90+CD49f lowCD62L-CD133+ subpopulation. Specifically, the expansion of megakaryocyte-biased HSCs is supported by a biomimetic niche-like microenvironment, which generates a suitable cytokine milieu and supplies the appropriate physical scaffolding. Thus, beyond clarifying the existence and immuno-phenotype of human megakaryocyte-biased HSC, our study demonstrates a flexible human HSC expansion strategy that could help realize the strong clinical promise of HSC-based therapies.

Heparin alleviates LPS-induced endothelial injury by regulating the TLR4/MyD88 signaling pathway.

Heparin is a commonly used in the clinic, however, Heparin's effect on endothelial injury remains unclear. The aim of the present study was to evaluate the effects and possible mechanisms of action underlying heparin treatment in lipopolysaccharide (LPS)-induced endothelial injury in vitro. TNF-α, IL-1ß, IL-6 and IFN-γ levels were measured using ELISA. Cell proliferation was measured using a 5-ethynyl-2'-deoxyuridine (EdU) assay. The number of apoptotic cells and apoptotic rate were evaluated using TUNEL assays and flow cytometry, respectively. Toll-like receptor 4 (TLR4), myeloid differentiation primary response 88 (MyD88) and NF-κB (p65) gene expression was evaluated using reverse transcription-quantitative PCR, whilst TLR4, MyD88 and p-NF-κB (p65) protein expression was evaluated using western blot analysis. The levels of phosphorylated NF-κB in the nucleus were evaluated using cellular immunofluorescence. Compared with those in the normal control group, TNF-α, IL-1ß, IL-6 and IFN-γ levels were significantly increased in the LPS group (P<0.001). In addition, 5-ethynyl-2'-deoxyuridine (EdU)-positive cells were significantly increased and apoptosis was significantly decreased (P<0.001). TLR4, MyD88 and NF-κB (p65) expression was also significantly increased (P<0.001). Compared with those in the LPS group, following heparin treatment, TNF-α, IL-1ß, IL-6 and IFN-γ levels were significantly decreased (P<0.05), whilst the number of EdU-positive cells was significantly increased and the level of apoptosis was significantly decreased (P<0.05). TLR4, MyD88 and NF-κB (p65) expression was also significantly decreased by heparin in a dose-dependent manner (P<0.001). Small interfering RNA-TLR4 transfection exerted similar effects to those mediated by heparin in alleviating endothelial injury. In conclusion, heparin suppressed LPS-induced endothelial injury through the regulation of TLR4/MyD88/NF-κB (p65) signaling in vitro.

Exendin-4 gene modification and microscaffold encapsulation promote self-persistence and antidiabetic activity of MSCs.

Mesenchymal stem cell (MSC)-based therapy to combat diabetic-associated metabolic disorders is hindered by impoverished cell survival and limited therapeutic effects under high glucose stress. Here, we genetically engineered MSCs with Exendin-4 (MSC-Ex-4), a glucagon-like peptide-1 (GLP-1) analog, and demonstrated their boosted cellular functions and antidiabetic efficacy in the type 2 diabetes mellitus (T2DM) mouse model. Mechanistically, MSC-Ex-4 achieved self-augmentation and improved survival under high glucose stress via autocrine activation of the GLP-1R-mediated AMPK signaling pathway. Meanwhile, MSC-Ex-4-secreted Exendin-4 suppressed senescence and apoptosis of pancreatic ß cells through endocrine effects, while MSC-Ex-4-secreted bioactive factors (e.g., IGFBP2 and APOM) paracrinely augmented insulin sensitivity and decreased lipid accumulation in hepatocytes through PI3K-Akt activation. Furthermore, we encapsulated MSC-Ex-4 in 3D gelatin microscaffolds for single-dose administration to extend the therapeutic effect for 3 months. Together, our findings provide mechanistic insights into Exendin-4-mediated MSCs self-persistence and antidiabetic activity that offer more effective MSC-based therapy for T2DM.

Comparison of Chondrocytes in Knee Osteoarthritis and Regulation by Scaffold Pore Size and Stiffness.

In knee osteoarthritis (OA), there is more pronounced cartilage damage in the medial compartment ("lesion zone") than the lateral compartment ("remote zone"). This study fills a gap in the literature by conducting a systematic comparison of cartilage and chondrocyte characteristics from these two zones. It also investigates whether chondrocytes from the different zones respond distinctly to changes in the physical and mechanical microenvironment using three-dimensional porous scaffolds by changing stiffness and pore size. Cartilage was harvested from patients with end-stage varus knee OA. Cartilage from the lesion and remote zones were compared through histological and biomechanical assessments, and through proteomic and gene transcription analyses of chondrocytes. Gelatin scaffolds with varied pore sizes and stiffness were used to investigate in vitro microenvironmental regulation of chondrocytes from the two zones. Cartilage from the lesion and remote zones differed significantly (p < 0.05) in histological and biomechanical characteristics, as well as phenotype, protein, and gene expression of chondrocytes. Chondrocytes from both zones were sensitive to changes in the structural and mechanical properties of gelatin scaffolds. Of interest, although all chondrocytes better retained chondrocyte phenotype in stiffer scaffolds, those from the lesion and remote zones, respectively, preferred scaffolds with larger and smaller pores. Distinct variations exist in cartilage and chondrocyte characteristics in the lesion and remote zones of knee OA. Cells in these two zones respond differently to variations in the physical and mechanical microenvironment. Understanding and manipulating these differences will facilitate the development of more efficient and precise diagnostic and therapeutic approaches for knee OA.

Synthetic liver fibrotic niche extracts achieve in vitro hepatoblasts phenotype enhancement and expansion.

It is still a challenge for synthesizing 'cellular niche-mimics' in vitro with satisfactory reproducibility and fidelity to recreate the natural niche components (e.g., extracellular matrices and soluble factors) for stem cell cultivation. Inspired by the massive amplification of hepatic progenitor cells during liver fibrosis in vivo, here we optimized the in vitro liver fibrotic niches and subsequently harvested their bioactive ingredients as niche extracts (NEs). The fibrosis-relevant NE marginally outperformed Matrigel for phenotype maintenance of human embryonic stem cell (hESC)-derived hepatoblasts (HBs) and recapitulation of the pathological angiogenesis of hESC-derived endothelial cells both in 2D culture and 3D liver organoids. Finally, defined NE components (i.e., collagen III, IV, IL-17, IL-18 and M-CSF) were resolved by the quantitative proteomics which exhibited advantage over Matrigel for multi-passaged HB expansion. The pathology-relevant and tissue-specific NEs provide innovative and generalizable strategies for the discovery of optimal cellular niche and bioactive niche compositions.

[Mechanism study of adaptive response in high background radiation area of Yangjiang in China].

OBJECTIVE: To study the adaptive response mechanisms in high background radiation area (HBRA) among Yangjiang local people through gene and protein expression of receptor for advanced glycation end products (RAGE) and S100A6 in peripheral blood and sputum in inhabitants of HBRA. METHODS: A total of 53 male inhabitants were selected from HBRA in Yangjiang as the exposure group, while 53 male inhabitants were selected from Enping (control area, CA)as the control group. The content of RAGE and S100A6 gene and protein were detected by RT-PCR and Western blotting assay. Thermo luminescent dosemeter(TLD) assay was used to measure the outside dose and estimate the effective dose. RESULTS: The effective dose in CA and HBRA was respectively 1.95 mSv and 6.24 mSv, which was 3 fold difference. Compared with CA, RAGE and S100A6 expression were significantly reduced in both gene and protein level in HBRA. The relative median mRNA expression of RAGE and S100A6 in peripheral blood were respectively 0.28, 1.06 and 0.16, 0.79 in CA and HBRA group, there was significance (with analysis Z values of -2.587 and -2.328 respectively, P < 0.05) with Wilcoxon rank test. For the protein of sputum, the relative median expression were respectively 2.98, 2.25 and 0.53, 0.47 with significant difference (with analysis Z values of -2.201 and -2.366 respectively, P < 0.05) by Wilcoxon rank test. CONCLUSION: The low expression of RAGE and S100A6 in HBRA group might be correlated with the adaptive response and the low mortality of cancer in HBRA.

Cryotherapy Models and Timing-Sequence Recovery of Exercise-Induced Muscle Damage in Middle- and Long-Distance Runners.

CONTEXT: Among sports-recovery methods, cold-water immersion (CWI), contrast-water therapy (CWT), and whole-body cryotherapy (WBC) have been applied widely to enhance recovery after strenuous exercise. However, the different timing effects in exercise-induced muscle damage (EIMD) after these recovery protocols remain unknown. OBJECTIVE: To compare the effects of CWI, CWT, and WBC on the timing-sequence recovery of EIMD through different indicator responses. DESIGN: Crossover study. SETTING: Laboratory. PATIENTS OR OTHER PARTICIPANTS: Twelve male middle- and long-distance runners from the Beijing Sport University (age = 21.00 ± 0.95 years). INTERVENTION(S): Participants were treated with different recovery methods (control [CON], CWI, CWT, WBC) immediately postexercise and at 24, 48, and 72 hours postexercise. MAIN OUTCOME MEASURE(S): We measured perceived sensation using a visual analog scale (VAS), plasma creatine kinase (CK) activity, plasma C-reactive protein (CRP) activity, and vertical-jump height (VJH) pre-exercise, immediately postexercise, and at 1, 24, 48, 72, and 96 hours postexercise. RESULTS: For the VAS score and CK activity, WBC exhibited better timing-sequence recovery effects than CON and CWI (P < .05), but the CWT demonstrated better effects than CON (P < .05). The CRP activity was lower after WBC than after the other interventions (P < .05). The VJH was lower after WBC than after CON and CWI (P < .05). CONCLUSIONS: The WBC positively affected VAS, CK, CRP, and VJH associated with EIMD. The CWT and CWI also showed positive effects. However, for the activity and timing-sequence effect, CWT had weaker effects than WBC.

High throughput scaffold-based 3D micro-tumor array for efficient drug screening and chemosensitivity testing.

Oncology drug development is greatly hampered by inefficient drug screening using 2D culture. Herein, we present ready-to-use micro-scaffolds in 384-well format to generate uniform 3D micro-tumor array (3D-MTA, CV < 0.15) that predicts in vivo drug responses more accurately than 2D monolayer. 3D-MTA generated from both cell lines and primary cells achieved high screen quality (Z' > 0.5), and were compatible with standard high throughput and high content instruments. Doxorubicin identified by 3D-MTA and 2D successfully inhibited tumor growth in mice bearing lung cancer cell line (H226) xenografts, but not gemcitabine and vinorelbine, which were selected solely by 2D. Resistance towards targeted therapy was modeled on 3D-MTA, which elicited SK-BR-3 to express higher proliferation-related genes in response to gefitinb, as compared to 2D. Screening of 56 MAPK inhibitors identified pisamertib to synergistically improve cytotoxicity effect in combination with gefitinib. Primary tumor cells derived from patient-derived xenografts further attested concordance of drug response in 3D-MTA with in vivo response. 3D-MTA was further extended to realize chemosensitivity testing using patient-derived cells. Overall, 3D-MTA demonstrated strong potential to accelerate drug discovery and improve cancer treatment by providing efficient drug screening.

The efficacy and safety of anti-fibrinolytic agents in blood management following peri-acetabular osteotomy: A meta-analysis.

BACKGROUND: Blood management after peri-acetabular osteotomy (PAO) has become a serious problem. We performed a meta-analysis to evaluate the efficacy and safety of antifibrinolytics for blood management after PAO. METHODS: PubMed, OVID, Embase, ScienceDirect, and Web of Science were searched up to January, 2018 without restrictions on publication date and language. We also searched the relevant publication sources. The research was reported according to the Preferred Reporting Items for Systematic Reviews and Meta-analysis guidelines. Randomized controlled trials (RCTs) and non-RCTs were included in our study. Weighted mean differences, risk difference, and 95% confidence intervals were calculated. We assessed statistical heterogeneity for each outcome with the use of a standard chi-square test and I statistic. The data were extracted by 2 of the co-authors independently and were analyzed by RevMan5.3. Primary outcomes were total blood loss, postoperative hemoglobin decline, and transfusion rates. Secondary outcomes were length of a hospital stay and postoperative complications. RESULTS: Four studies including 1 RCT and 3 non-RCTs were included in our study. The present meta-analysis indicated that antifibrinolytics was associated with a significant reduction of the total blood loss, postoperative hemoglobin decline, transfusion rates, and length of a hospital stay compared with control groups. No significant differences were identified in terms of the incidence of postoperative complications. CONCLUSION: Intravenous antifibrinolytics was efficacious in reduction of total blood loss, postoperative hemoglobin decline, and length of a hospital stay after PAO without increasing the risk of thromboembolic complications. More high-quality RCTs with long follow-up period were necessary for proper comparisons of the efficacy and safety of antifibrinolytics with placebo.

Serum zinc is associated with plasma leptin and Cu-Zn SOD in elite male basketball athletes.

This paper investigates the relationship between plasma trace element and plasma leptin, as well as percent fat mass, in 16 male basketball athletes. Blood samples were obtained before intensive training and 24h after intensive training to measure plasma zinc (Zn), copper (Cu), calcium (Ca), magnesium (Mg), iron (Fe), and leptin levels. High-density lipoprotein cholesterol (HDL), low-density lipoprotein cholesterol (LDL), triglyceride (TG), total and cholesterol (TC) levels were determined using commercially available kits for humans. Subjects presented similar values in terms of age (21.1±2.2 years old), body mass index (23.9±2.00kg/m(2)), percent body fat (14.40±1.52%), plasma hemoglobin (150.1±9.4g/L), plasma Zn (17.47±1.28µmol/l), plasma Cu (13.42±1.40µmol/L), plasma Ca (2.41±0.14mmol/L), and plasma Mg (0.96±0.02mmol/L). The correlation analysis between degree of plasma leptin and plasma element contents was performed using the SPSS 16.0 software. Plasma Zn correlated positively with plasma leptin (r=0.746, P<0.01), Cu-Zn SOD (r=0.827, P<0.01), and negatively with percent fat mass (r=-0.598, P<0.05) under no-training conditions. Meanwhile, plasma Cu, Ca, Mg, and Fe did not correlate with plasma leptin or percent fat mass (P>0.05). In conclusion, plasma Zn may be involved in the regulation of plasma leptin and may serve as a lipid-mobilizing factor in Chinese men's basketball athletes.

Up-regulation of RAGE and S100A6 in rats exposed to cigarette smoke.

Cigarette smoke has been widely investigated in terms of epidemiology and pathological endpoints in relation to human lung diseases and animal study. In this study we exposed Wistar rats to cigarette smoke at concentrations of 20% and 60% to explore potential molecular mechanisms at the protein level. Exposures were conducted twice a day, 5 days a week for 43 weeks. As a major metabolite of nicotine in cigarette, cotinine level in rat urine was determined by HPLC-MS. A dose-dependent analysis indicated that cotinine may be used as an exposure marker of cigarette smoke. Expression of receptor for advanced glycation endproducts (RAGE), an immunoglobulin super family that triggers the intracellular signal cascade reaction leading to inflammation and its ligand S100A6 (calgranulin) in bronchial epithelial cells and lung tissues of rats, were found to be positive correlated with cotinine levels, indicating that RAGE and S100A6 may be attributable to inflammation and oxidative damage caused by cigarette smoke.