Objective:To investigate the inhibitory effect and mechanisms of action of mucopolysaccharide polysulfate cream on hypertrophic scar formation.Methods:Circular full-thickness wounds with a diameter of 6 mm were made in both ears of 16 New Zealand white rabbits to establish a rabbit ear model of hypertrophic scar. There were 3 hypertrophic scars in each rabbit ear. About 14 days after the operation, scars on the left ear were topically treated with mucopolysaccharide polysulfate cream, and served as the experimental group; scars on the right ear were topically treated with the cream vehicle, and served as vehicle control group. The dosage of topical agents for one rabbit ear was approximately 0.4 g, which were given twice a day for 6 consecutive weeks. Scar tissues were collected on days 0, 14 and 42, that is, 14, 28 and 56 after operation respectively, and subjected to hematoxylin and eosin (HE) staining, Masson staining and immunohistochemical study, so as to evaluate histopathological scores, measure the scar thickness and collagen fiber density, and determine the expression of type Ⅰ and Ⅲ collagen and the ratio of type Ⅰ/Ⅲ collagen. The t test and one-way analysis of variance were used to compare the indices between groups. Results:Compared with pretreatment histopathological manifestations, HE staining showed extensive extracellular matrix deposition, inflammatory cell infiltration and local hyperemia in the control group after 42-day treatment, but no obvious changes in the experimental group. The pathological scores of scar tissues on the rabbit ears significantly increased over time in the control group (days 0, 14 and 42: 4.16 ± 1.61, 6.50 ± 1.46, 6.53 ± 1.34, respectively; F = 13.69, P = 0.001) , while there was no significant change in the experimental group (days 0, 14 and 42: 4.65 ± 1.52, 5.13 ± 1.83, 5.38 ± 1.60, respectively; F = 0.78, P > 0.05) . Masson staining showed extremely high content of dark blue-dyed collagen fibers in the control group on day 42, but there was a decrease in the content of collagen fibers in the experimental group; with the increase in treatment duration, the thickness of scar tissues significantly increased in the control group compared with that before treatment ( F = 5.64, P = 0.007) , while there was no significant change in the experimental group ( F = 1.48, P > 0.05) . Immunohistochemical study revealed no significant change in the expression of type Ⅲ collagen in either the experimental group or the control group at any of the above posttreatment time points compared with that on day 0 ( F = 0.22, 0.92, respectively, both P > 0.05) , but the expression of type Ⅰ collagen and the ratio of type Ⅰ/Ⅲ collagen significantly increased in the control group ( F = 7.47, P < 0.001; F = 4.70, P = 0.005, respectively) . On day 42, the expression of type Ⅰ collagen and the ratio of type Ⅰ/Ⅲ collagen significantly decreased in the experimental group compared with the control group ( t = 3.04, P = 0.007; t = 2.35, P = 0.030, respectively) . Conclusion:Topical mucopolysaccharide polysulfate cream is effective in preventing and inhibiting scar hypertrophy by reducing the scar thickness and inhibiting the collagen fiber hyperplasia and type I collagen expression.

Objective:To investigate the protective effect of ulinastatin on sepsis-acute kidney injury (SA-AKI) by NF-κB signaling pathway.Methods:Total of 60 mice were randomly(random number) divided into sham group, cecal ligation puncture group (CLP group) and ulinastatin treatment group (CLP+UTI group). Ulinastatin treatment group was intraperitoneally injected with ulinastatin 50 000 U/kg once a day. 24 hours after operation, five mice were sacrificed, the kidney tissues were collected to observe renal histopathology by HE staining. The macrophage infiltration was observed by immunohistochemistry. The remaining mice in each group were used to calculate the survival rate of 7-day after operation. HK-2 cells were stimulated by LPS to obtain the SA-AKI model, and the cells were divided into control group, LPS group and LPS + UTI group. CCK-8 assay was used to detect cell viability, EdU assay was used to detect cell proliferation, and JC-1 assay was used to detect mitochondrial damage. The phosphorylation degree of NF-κB was detected by western blot. Inflammatory factors concentrations of cellular supernatant were detected by ELISA assay.Results:Compared with the sham group, the kidney tissue of mice in CLP group showed that kidney pathological obvious changed, the infiltration of macrophages increased, and the survival rate of mice decreased. CLP+ UTI group reduced the pathological changes and the infiltration of macrophages, improved the survival rate of mice. Compared with control group, LPS group obviously inhibited the cells activity and proliferation of HK-2 cells, damaged the mitochondrial membrane potential of HK-2 cells. Compared with LPS group, LPS+ UTI group attenuated the phosphorylation of NF-κB, decreased the secretion of inflammatory factors, rescued the activity and proliferation of HK-2 cells, and reduced the damage of HK-2 mitochondrial membrane potential.Conclusions:Ulinastatin can reduce mitochondrial damage, inhibit the secretion of inflammatory factors and improve the function of renal tubular epithelial cells through regulating NF-κB signaling pathway.

BACKGROUND: Systemic lupus erythematosus (SLE) is a complex autoimmune disease, and the mechanism of SLE is yet to be fully elucidated. The aim of this study was to explore the role of two-pore segment channel 2 (TPCN2) in SLE pathogenesis. METHODS: Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expression of TPCN2 in SLE. We performed a loss-of-function assay by lentiviral construct in Jurkat and THP-1 cell. Knockdown of TPCN2 were confirmed at the RNA level by qRT-PCR and protein level by Western blotting. Cell Count Kit-8 and flow cytometry were used to analyze the cell proliferation, apoptosis, and cell cycle of TPCN2-deficient cells. In addition, gene expression profile of TPCN2-deficient cells was analyzed by RNA sequencing (RNA-seq). RESULTS: TPCN2 knockdown with short hairpin RNA (shRNA)-mediated lentiviruses inhibited cell proliferation, and induced apoptosis and cell-cycle arrest of G2/M phase in both Jurkat and THP-1 cells. We analyzed the transcriptome of knockdown-TPCN2-Jurkat cells, and screened the differential genes, which were enriched for the G2/M checkpoint, complement, and interleukin-6-Janus kinase-signal transducer and activator of transcription pathways, as well as changes in levels of forkhead box O, phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin, and T cell receptor pathways; moreover, TPCN2 significantly influenced cellular processes and biological regulation. CONCLUSION TPCN2 might be a potential protective factor against SLE.
Apoptosis/genetics* ; Cell Division ; Humans ; Jurkat Cells ; Lupus Erythematosus, Systemic/genetics* ; RNA, Small Interfering/genetics*

BACKGROUND: Pathological scars are a disorder that can lead to various cosmetic, psychological, and functional problems, and no effective assessment methods are currently available. Assessment and treatment of pathological scars are based on cutaneous manifestations. A two-photon microscope (TPM) with the potential for real-time non-invasive assessment may help determine the under-surface pathophysiological conditions in vivo . This study used a portable handheld TPM to image epidermal cells and dermal collagen structures in pathological scars and normal skin in vivo to evaluate the effectiveness of treatment in scar patients. METHODS: Fifteen patients with pathological scars and three healthy controls were recruited. Imaging was performed using a portable handheld TPM. Five indexes were extracted from two dimensional (2D) and three dimensional (3D) perspectives, including collagen depth, dermo-epidermal junction (DEJ) contour ratio, thickness, orientation, and occupation (proportion of collagen fibers in the field of view) of collagen. Two depth-dependent indexes were computed through the 3D second harmonic generation image and three morphology-related indexes from the 2D images. We assessed index differences between scar and normal skin and changes before and after treatment. RESULTS: Pathological scars and normal skin differed markedly regarding the epidermal morphological structure and the spectral characteristics of collagen fibers. Five indexes were employed to distinguish between normal skin and scar tissue. Statistically significant differences were found in average depth ( t = 9.917, P <0.001), thickness ( t = 4.037, P <0.001), occupation ( t = 2.169, P <0.050), orientation of collagen ( t = 3.669, P <0.001), and the DEJ contour ratio ( t = 5.105, P <0.001). CONCLUSIONS Use of portable handheld TPM can distinguish collagen from skin tissues; thus, it is more suitable for scar imaging than reflectance confocal microscopy. Thus, a TPM may be an auxiliary tool for scar treatment selection and assessing treatment efficacy.
Humans ; Cicatrix/diagnostic imaging* ; Skin/pathology* ; Collagen ; Imaging, Three-Dimensional/methods*