Objective To investigate the expression of autophagy and the effect of complement C5a/C5aR pathway on autophagy induced by renal ischemia reperfusion injury (IRI).Methods MaleWT and C5aR gene knockout (BALB/C background) mice were selected.The model of renal IRI was established by occluding bilateral renal pedicles with microaneurysm clamps.Mice were divided into wild type BALB/C (WT) group and C5aR gene knock out (C5aRKO) group.The pathology of kidney was assessed by HE staining.The levels of BUN and KIM-1 were detected 24 h after reperfusion.The expression of the autophagy-associated protein (LC3 Ⅱ/LC3 Ⅰ and P62) was measured by Western blotting and immunofluorescence.In vitro,human renal tubular epithelial cells (HK2) were cultured.The expression of LC3 in HK2 cells was investigated by immunofluorescence and Western blotting after being treated with recombinant C5a or C5a combined with C5aR antagonist (C5aRA).Results As compared with WT group,the severity of kidney injury was obviously reduced in C5aRKO group (P＜0.05).After ischemia-reperfusion,the expression of autophagy-related protein LC3 gradually increased with the reperfusion time prolonged.The level of autophagy induced by ischemia-reperfusion was significantly reduced in C5aRKO group as compared with WT group (P＜0.05).In addition,the expression of autophagy-related protein LC3 Ⅱ in HK2 cells was increased with the augment of C5a stimulation concentration in vitro.Blockage of C5aR pathway by C5aRA led to a significant decrease in autophagy (P ＜ 0.05).Conclusion Complement C5a/C5aR pathway promotes renal IRI-induced autophagy.

Objective To investigate the expression of proinflammatory cytokine interleukin-17(IL-17) and complement cleavage fragment C5a and the regulation effect of C5a on the expression of IL-17 during renal allograft rejection .Methods The frequency of IL-17+ T cell in peripheral blood and the expression of IL-17 in renal tubular epithelial cells (HK2) after C5a stimulation in renal transplant recipients were measured by flow cytometry and the changes of serum C5a level was detected by ELISA ,respectively .Im-munohistochemistry was applied to detect and compare the expression of IL-17 and the deposition of C5b-9 in normal renal tissues and renal tissues with allograft rejection .The difference of IL-17 expression in HK2 cells before and after the recombinant C5a stimulation was detected by immunocytochemistry .Results Both the percentage of IL-17+ T cells and serum C5a levels were sig-nificantly increased after the allogeneic renal transplantation .Compared with normal renal tissues ,both the deposition of C5b-9 and the IL-17 expression in renal tissues with allograft rejection was remarkably up-regulated ,which showed the positive correlation be-tween them .The expression of IL-17 in HK2 was obviously up-regulated by the recombinant C5a stimulation .Conclusion C5a may positively regulate the expression of IL-17 by tubular epithelial cells during the renal allograft rejection .

BACKGROUND:Cel co-culture can maximize the simulation ofin vivomicroenvironment. Cel scratch test and interleukin-1β can destroy the balance between matrix metaloproteinases (MMPs) and matrix metaloproteinase inhibitors (TIMPs), resulting in extracelular matrix degradation of the articular cartilage, functional disorders of chondrocytes and articular cartilage degeneration. OBJECTIVE:To study the effect of interleukin-1β on migration, MMP and TIMP expression of chondrocytes co-cultured with osteoblast supernatantin vitro. METHODS:There were three groups: chondrocyte monoculture group, osteoblast+chondrocyte group (co-culture group), osteoblast+chondrocyte+interleukin-1β group (interleukin-1β group). Cel scratch test was conducted to observe the migration of chondrocytes within 24 hours. Semi-quantitative PCR test was used to detect the changes in expressions of MMP-1, MMP-2, MMP-3, MMP-9, TIMP-1, TIMP-2, TIMP-3, TIMP-9 in chondrocytes within 24 hours. RESULTS AND CONCLUSION:Compared with the monoculture group, cel migration rate of the other two groups were increased significantly (P< 0.01). Compared with the monoculture group, the gene expressions of MMP-1, MMP-2, MMP-3 and MMP-9 were increased significantly in the coculture group (P < 0. 05); the gene expressions of MMP-1, MMP-3, MMP-9 were increased significantly in the interleukin-1β group (P< 0. 01). Compared with monoculture group, the gene expression of TIMP-1 was increased significantly (P < 0. 01), but the gene expressions of TIMP-3 and TIMP-4 were declined significantly (P < 0. 05) in the other two groups. These findings indicate that co-culture of chondrocytes with osteoblasts can promote chondrocytes migration, enhance gene expression of chondrocytes MMP-1, MMP-2, MMP-3, MMP-9 and regulate gene expression of TIMPs family. Interleukin-1β inhibitsthe migration of chondrocytes co-cultured with osteoblasts and gene expression of TIMPs family.

Generally, extracellular matrix (ECM) has the characteristics of viscoelasticity. In osteoarthritis (OA), catabolic processes alter the viscoelastic properties of functional pericellular matrix (PCM) of chondrocytes. Chondrocytes sense and respond to their mechanical microenvironment via an array of mechanosensitive receptors and channels that activate a complex network of downstream signaling pathways to regulate several cell processes central to OA pathology. Advances in understanding the specific mechanosignalling mechanisms in articular cartilage will promote the development of cell microenvironment construction in cartilage tissue engineering and the targeted precision therapeutics for OA. In this review, the work on the mechanism of matrix viscoelasticity regulating chondrocytes mechanotransduction by Agarwal et al. was briefly commented, and the recent advances related with their work was also discussed.

This study was designed to investigate the effects of TNFSF14 on mitochondrial function in ischemia/reperfusion-induced acute kidney injury(I/R-AKI)and its mechanism.TNFSF14-/-and TNFSF14+/+mice underwent renal ischemia-reperfusion operation to establish I/R-AKI models,and their histopathology changes were compared by using Periodic Acid-Schiff stain,transmission electron microscopy.Immunohistochemistry(IHC)was used to detect levels of TNFSF14,HVEM,LT[3R,mitochondrial activity related proteins(Dpr1 and Mfn2)and inflammatory cells infiltration in kidney tissues of mice and relevant patients.In vitro cell experiments,immunofluorescence and immunoblotting were used to observe the effects of exogenous recombinant TNFSF14 factor on the damage and mitochondrial activity of renal tubular epithelial cell line HK-2 cells.Data showed that the expression of TNFSF14 and its receptors were significantly increased in kidney tissues of I/R-AKI mice and clinical human renal tissues of acute tubular injury.Compared with the sham group,I/R mice showed significantly higher levels of renal tubular injury score,inflammatory cells infiltration,cell apoptosis,mitochondrial damage,and Drp1 expression,while knocking out the TNFSF14 gene,the above indicators were significantly reduced.In vitro,exogenous TNFSF14 stimulation could aggravate the hypoxia-induced apoptosis and the decrease of mitochondrial membrane potential in HK-2 cells,while increasing phosphorylation of Drp1 at Se616 to promote its transfer from cytoplasm to mitochondria,leading to an abnormal increase in mitochondrial fission.In conclusions,TNFSF14 may mediate the pathological process of I/R-AKI by promoting mitochondrial damage.

Objective:To observe the clinical effect of ablative fractional CO 2 laser in the treatment of hypertrophic scar in children after burn. Methods:The clinical data of patients with post-burn hypertrophic scar in children who met the inclusion and exclusion criteria in the Cosmetic Clinic and Burn Clinic of the Third People’s Hospital of Bengbu City Affiliated to Bengbu Medical College from January 2019 to March 2021 were collected, and a retrospective study was conducted. All patients were divided into laser group and control group, and laser group was further divided into 1-2 times subgroups and 3-4 times subgroups. The laser group received ablative fractional CO 2 laser treatment 1-4 times on the basis of conventional anti-scar treatment (pressure therapy and topical silicone drugs), and the treatment interval was 1-3 months; the control group only received conventional anti-scar treatment. The color, blood vessel distribution, thickness and softness of scar were scored by Vancouver Scar Scale (VSS), before treatment and 2 months after treatment In the laser group, and at 3 and 6 months in the control group, respectively. The degree of pruritus of the scar was scored with Visual Analogue Scale (VAS). The patient’s satisfaction evaluation is graded as four levels: very satisfied, relatively satisfied, generally satisfied, and dissatisfied. All data were analyzed by SPSS 19.0 software with paired t-test, Wilcoxon rank-sum test or chi-square test according to the type and nature of the data. Results:A total of 103 patients with hypertrophic scars were included, with a total of 134 scars, including 58 males and 45 females; the age was (3.9±3.0) years, range 0-11 years old; the scar area accounted for 4.2%±3.1% of the body surface area; the course of scar was (3.6±2.2) months. There were 72 patients (94 scars) in the laser group, including 29 patients (37 scars) in the 1-2 times subgroup and 43 (57 scars) in the 3-4 times subgroup; 31 patients in the control group(40 scars). (1) Vascular distribution, softness and overall score assessed by VSS in 1-2 times subgroup after treatments were significantly lower than those before treatment ( P<0.05). The thickness, blood vessel distribution, softness and overall score assessed by VSS in 3-4 times subgroup after treatments were significantly lower than those before treatment ( P<0.05). The improvement degree of scar after treatment in each group was different. Compared with the control group, the improvement degree in the 1-2 times subgroup and the 3-4 times subgroup was more obvious ( P<0.05). The improvement degree in the 3-4 times subgroup was better than that in the 1-2 times subgroups ( P<0.05). (2) Compared with before treatment, the VSS scores of scars after laser treatment in different parts were significantly different except for the thickness scores of face and neck, trunk, and upper limbs group ( P<0.05). (3) The degree of pruritus was assessed by VAS method. The pruritus score of the 1-2 times subgroups and 3-4 times subgroups before treatment was (4.86±1.35) points, (4.97±0.93) points, and the pruritus score 2 months after treatment was (1.93±0.99) points, (1.90±0.83) points, the pruritus score improved significantly after treatment, and the difference was statistically significant ( P<0.01). The pruritus scores of the control group at 3 months and 6 months were (4.83±0.82) points and (4.22±0.66) points, and the scores at 6 months were slightly improved compared with those at 3 months, and the difference was statistically significant ( P<0.01). (4) In the laser group, 5 patients (6.9%) had pigmentation after the first treatment, and then gradually subsided; 7 patients (9.7%) had blisters after the second treatment, which healed after dressing change. In the control group, 3 cases (9.7%) had erosions, and the erosions improved after adjusting the pressure appropriately. (5) The overall satisfaction of patients in the laser group was higher than that in the control group [100% (72/72) vs. 80.6% (25/31), P<0.05]. Conclusions:Fractional CO 2 laser has a good effect on the treatment of hypertrophic scars in early childhood burns. It can effectively inhibit scar hyperplasia and improve the degree of itching in patients. The satisfaction of both doctors and patients is high.

Objective:To observe the clinical effect of ablative fractional CO 2 laser in the treatment of hypertrophic scar in children after burn. Methods:The clinical data of patients with post-burn hypertrophic scar in children who met the inclusion and exclusion criteria in the Cosmetic Clinic and Burn Clinic of the Third People’s Hospital of Bengbu City Affiliated to Bengbu Medical College from January 2019 to March 2021 were collected, and a retrospective study was conducted. All patients were divided into laser group and control group, and laser group was further divided into 1-2 times subgroups and 3-4 times subgroups. The laser group received ablative fractional CO 2 laser treatment 1-4 times on the basis of conventional anti-scar treatment (pressure therapy and topical silicone drugs), and the treatment interval was 1-3 months; the control group only received conventional anti-scar treatment. The color, blood vessel distribution, thickness and softness of scar were scored by Vancouver Scar Scale (VSS), before treatment and 2 months after treatment In the laser group, and at 3 and 6 months in the control group, respectively. The degree of pruritus of the scar was scored with Visual Analogue Scale (VAS). The patient’s satisfaction evaluation is graded as four levels: very satisfied, relatively satisfied, generally satisfied, and dissatisfied. All data were analyzed by SPSS 19.0 software with paired t-test, Wilcoxon rank-sum test or chi-square test according to the type and nature of the data. Results:A total of 103 patients with hypertrophic scars were included, with a total of 134 scars, including 58 males and 45 females; the age was (3.9±3.0) years, range 0-11 years old; the scar area accounted for 4.2%±3.1% of the body surface area; the course of scar was (3.6±2.2) months. There were 72 patients (94 scars) in the laser group, including 29 patients (37 scars) in the 1-2 times subgroup and 43 (57 scars) in the 3-4 times subgroup; 31 patients in the control group(40 scars). (1) Vascular distribution, softness and overall score assessed by VSS in 1-2 times subgroup after treatments were significantly lower than those before treatment ( P<0.05). The thickness, blood vessel distribution, softness and overall score assessed by VSS in 3-4 times subgroup after treatments were significantly lower than those before treatment ( P<0.05). The improvement degree of scar after treatment in each group was different. Compared with the control group, the improvement degree in the 1-2 times subgroup and the 3-4 times subgroup was more obvious ( P<0.05). The improvement degree in the 3-4 times subgroup was better than that in the 1-2 times subgroups ( P<0.05). (2) Compared with before treatment, the VSS scores of scars after laser treatment in different parts were significantly different except for the thickness scores of face and neck, trunk, and upper limbs group ( P<0.05). (3) The degree of pruritus was assessed by VAS method. The pruritus score of the 1-2 times subgroups and 3-4 times subgroups before treatment was (4.86±1.35) points, (4.97±0.93) points, and the pruritus score 2 months after treatment was (1.93±0.99) points, (1.90±0.83) points, the pruritus score improved significantly after treatment, and the difference was statistically significant ( P<0.01). The pruritus scores of the control group at 3 months and 6 months were (4.83±0.82) points and (4.22±0.66) points, and the scores at 6 months were slightly improved compared with those at 3 months, and the difference was statistically significant ( P<0.01). (4) In the laser group, 5 patients (6.9%) had pigmentation after the first treatment, and then gradually subsided; 7 patients (9.7%) had blisters after the second treatment, which healed after dressing change. In the control group, 3 cases (9.7%) had erosions, and the erosions improved after adjusting the pressure appropriately. (5) The overall satisfaction of patients in the laser group was higher than that in the control group [100% (72/72) vs. 80.6% (25/31), P<0.05]. Conclusions:Fractional CO 2 laser has a good effect on the treatment of hypertrophic scars in early childhood burns. It can effectively inhibit scar hyperplasia and improve the degree of itching in patients. The satisfaction of both doctors and patients is high.

Extracellular matrix (ECM) has been implicated in tumor progress and chemosensitivity. Ovarian cancer brings a great threat to the health of women with a significant feature of high mortality and poor prognosis. However, the potential significance of matrix stiffness in the pattern of long non-coding RNAs (lncRNAs) expression and ovarian cancer drug sensitivity is still largely unkown. Here, based on RNA-seq data of ovarian cancer cell cultured on substrates with different stiffness, we found that a great amount of lncRNAs were upregulated in stiff group, whereas SNHG8 was significantly downregulated, which was further verified in ovarian cancer cells cultured on polydimethylsiloxane (PDMS) hydrogel. Knockdown of SNHG8 led to an impaired efficiency of homologous repair, and decreased cellular sensitivity to both etoposide and cisplatin. Meanwhile, the results of the GEPIA analysis indicated that the expression of SNHG8 was significantly decreased in ovarian cancer tissues, which was negatively correlated with the overall survival of patients with ovarian cancer. In conclusion, matrix stiffening related lncRNA SNHG8 is closely related to chemosensitivity and prognosis of ovarian cancer, which might be a novel molecular marker for chemotherapy drug instruction and prognosis prediction.
Female ; Humans ; Cisplatin/pharmacology* ; Elasticity/physiology* ; Etoposide ; Extracellular Matrix/physiology* ; Ovarian Neoplasms/metabolism* ; RNA, Long Noncoding/metabolism*

Mechanical signal transduction are crucial for chondrocyte in response to mechanical cues during the growth, development and osteoarthritis (OA) of articular cartilage. Extracellular matrix (ECM) turnover regulates the matrix mechanical microenvironment of chondrocytes. Thus, understanding the mechanotransduction mechanisms during chondrocyte sensing the matrix mechanical microenvironment can develop effective targeted therapy for OA. In recent decades, growing evidences are rapidly advancing our understanding of the mechanical force-dependent cartilage remodeling and injury responses mediated by TRPV4 and PIEZOs. In this review, we highlighted the mechanosensing mechanism mediated by TRPV4 and PIEZOs during chondrocytes sensing mechanical microenvironment of the ECM. Additionally, the latest progress in the regulation of OA by inflammatory signals mediated by TRPV4 and PIEZOs was also introduced. These recent insights provide the potential mechanotheraputic strategies to target these channels and prevent cartilage degeneration associated with OA. This review will shed light on the pathogenesis of articular cartilage, searching clinical targeted therapies, and designing cell-induced biomaterials.
Chondrocytes ; TRPV Cation Channels ; Mechanotransduction, Cellular ; Biocompatible Materials ; Cartilage, Articular

Objective To fabricate a foldable microplate for single cell culture and establish finite element model of the folding microplate, so as to calculate traction force of single cells during contraction in three-dimensional (3D) state.Methods The folding angle of the microplate casued by cell traction force was calculated. Then the relation between bending moment and folding angle as well as the relation between traction force and bending moment were derived by using finite element simulation, so as to realize the characterization of traction force for singel cell in 3D state.Results The folding angles of the microplate with HSF and MC3T3-E1 cells in 3D state were 73°-173° and 49°-138°, respectively. The single cell traction forces of HSF and MC3T3-E1 cells were 55-210 nN and 52-161 nN, respectively.Conclusions The proposed method for measuring traction force of single cells in 3D state by fabricating the foldable microplate for single cell culture will provide some references for further development of calculating traction forces in 3D cell adhesion, spreading and migration.