Saikosaponin-d alleviates hepatic fibrosis through regulating GPER1/autophagy signaling

Background Hepatic fibrosis is the final pathway of chronic liver disease characterized by excessive accumulation of extracellular matrix (ECM), which eventually develop into cirrhosis and liver cancer. Emerging studies demonstrated that Saikosaponin-d (SSd) exhibits a protective role in liver fibrosis. However, the mechanism underlying anti-liver fibrosis of SSd in vivo and in vitro remains unclear. Methods and results Transforming growth factor (TGF)-β and carbon tetrachloride (CCl4) were used for creating liver fibrosis model in vitro and in vivo, respectively. The role of SSd in regulating liver fibrosis was assessed through Sirius red and Masson staining, and IHC assay. We found that SSd attenuated remarkably CCl4-induced liver fibrosis as evidenced by decreased collagen level, and decreased expression of fibrotic markers Col 1 and α-SMA. Meanwhile, SSd repressed autophagy activation as suggested by decreased BECN1 expression and increased p62 expression. Compared with HSCs from CCl4-treated group, the primary HSCs from SSd-treated mice exhibited a marked inactivation of autophagy. Mechanistically, SSd treatment enhanced the expression of GPER1 in primary HSCs and in TGF-β-treated LX-2 cells. GPER1 agonist G1 repressed autophagy activation, whereas GPER1 antagonist G15 activated autophagy and G15 also damaged the function of SSd on suppressing autophagy, leading to subsequent increased levels of fibrotic marker level in LX-2 cells. Conclusions Our findings highlight that SSd alleviates hepatic fibrosis by regulating GPER1/autophagy pathway.

Effects of Cryofrequency at Dermal and Hypodermic Level: Case Study

Background: Cryofrequency consists of the combination of cryotherapy and radiofrequency, acting simultaneously in the treatment of flaccidity and localized adiposity. Objective: To investigate the effects of cryofrequency at the dermal and hypodermic levels. Materials and methods: This is a case study, composed of one woman of 39-year-old, with interest to perform abdominoplasty surgery, presenting adiposity and flaccidity located in infra-umbilical region. Its treatment consists of eight cryofrequence applications. Results: There was reduction of 7 cm on supra-umbilical plicometry and 6 cm on infra-umbilical; a decrease of 2.7 kg of body weight. The ultrasound data show a decrease of 0.6 mm, in the thickness of the adipose layer. In the histological analysis: thicker epidermis, with more layers and dermis with greater number of fibroblasts and inflammatory cells, greater quantity of neoformed collagen tissue. Conclusion: The application of cryofrequence promoted reduction of adipose tissue and increased production of collagen. Level of Evidence: Level III, case-control study.

Saikosaponin-d Alleviates Hepatic Fibrosis Through Regulating GPER1/autophagy Signaling

Hepatic fibrosis is the final pathway of chronic liver disease characterized by excessive accumulation of extracellular matrix (ECM), which eventually develop into cirrhosis and liver cancer. Emerging studies demonstrated that Saikosaponin-d (SSd) exhibits a protective role in liver fibrosis. The current study mainly explored the mechanism underlying anti-liver fibrosis of SSd in vivo and in vitro. Transforming growth factor (TGF)-β and carbon tetrachloride (CCl4) were used for creating liver fibrosis model in vitro and in vivo, respectively. The role of SSd in regulating liver fibrosis was assessed through Sirius red and Masson staining, and IHC assay. We found that SSd attenuated remarkably CCl4-induced liver fibrosis as evidenced by decreased collagen level, and decreased expression of fibrotic markers Col 1 and α-SMA. Meanwhile, SSd repressed autophagy activation as suggested by decreased BECN1 expression and increased p62 expression. Compared with HSCs from CCl4-treated group, the primary HSCs from SSd-treated mice exhibited a marked inactivation of autophagy. Mechanistically, SSd treatment enhanced the expression of GPER1 in primary HSCs and in TGF-β-treated LX-2 cells. GPER1 agonist G1 repressed autophagy activation, whereas GPER1 antagonist G15 activated autophagy and G15 also damaged the function of SSd on suppressing autophagy, leading to subsequent increased levels of fibrotic marker level in LX-2 cells. Collectively, our findings highlight that SSd alleviates hepatic fibrosis by regulating GPER1/autophagy pathway.

Porous matrix nanometric scaffold repairs rabbit tibia injury by promoting bone marrow mesenchymal stem cell osteogenic differentiation and up-regulating type I and II collagen content

To explore the effect of porous matrix nanometer scaffold on Bone marrow mesenchymal stem cell osteogenic differentiation and type I and II collagen content in rabbit tibial injury. 22 rabbits, weighing 2.20–3.10 kg, were equally divided into model and treatment groups. Von Kossa was used to detect osteogenic differentiation, while Western blot detected type I and type II collagen. The bone density meter measurement software was used to assess tibial bone density. Universal material testing machine was used to detect biomechanical indicators level, while HE staining detected bone morphology. Adherent Cells growth were observed in a week; where cells clusters appeared and continued to grow 2 weeks later, and the number of osteoblasts and mineralized nodules increased significantly 3 weeks later. The collagen level in the treatment group was significantly higher than that in model group (P < 0.05). The tibia bone mineral density in model group was significantly lower at the sixth and twelfth weeks after surgery (P < 0.05). The maximum bending load and bending stiffness of model group were significantly lower than those of treatment group with higher damage degree (P < 0.05). Treatment increased the bone tissue and osteoblasts, leading to production of trabecular bone structure. No clustered regenerated bone-like tissues in model group were observed. The porous matrix nanometer scaffold can repair rabbit tibia injury possibly by promoting Bone marrow mesenchymal stem cell osteogenic differentiation and increasing type I and type II collagen level.

A Rationale for Using Sonoelastography in Thyroid Nodular Pathology

Objective. To provide a rationale for using sonoelastography (SEG) in the differential diagnosis of thyroid cancer (TC).Material and methods. Thirty patients with thyroid nodules of various morphological structures were examined. The authors studied the data of SEG and immunohistochemistry (IHC) with monoclonal antibodies against types III and IV collagen (they evaluated the degree of the expressed collagen fibers). Analysis of variance, ROC analysis, and logistic regression were used (by comparing with the expression of collagens) to assess the predictive ability of ultrasound.Results. The study showed that irregular and uneven contours, microcalcifications, and “the height greater than the width” were most significant among the ultrasound signs in the diagnosis of TC. Cool colors prevailed when performing SEG in the pattern of thyroid cancer. Purple-blue hues were predominantly recorded (p<0.05 with regard to benign nodules), green ones were less frequently. ROC analysis of compression elastography showed that the area under the curve was 0.785 (95% CI 0.740-0.826), sensitivity 78.1%, specificity 79.0%. Comparison of the data of IHC and SEG revealed a direct correlation of tissue elasticity with the degree of a stromal component and with the presence of collagen-containing structures.Conclusion. SEG may suppose the probable nature of thyroid nodules on the basis of their morphological features. The low degree of the stromal component and the low content of types III and IV collagen make follicular colloid goiter and adenoma soft, which is recorded at SEG. TC is characterized by a high collagen level attributable to the characteristics of the metabolism of cancer cells, which makes them solid in the mode of SEG.

Early pathological signs in young dysf−/− mice are improved by halofuginone

Dysferlinopathies are a non-lethal group of late-onset muscular dystrophies. Here, we evaluated the fusion ability of primary myoblasts from young dysf−/− mice and the muscle histopathology prior to, and during early stages of disease onset. The ability of primary myoblasts of 5-weekold dysf−/− mice to form large myotubes was delayed compared to their wild-type counterparts, as evaluated by scanning electron microscopy. However, their fusion activity, as reflected by the presence of actin filaments connecting several cells, was enhanced by the antifibrotic drug halofuginone. Early dystrophic signs were already apparent in 4-week-old dysf−/− mice; their collagen level was double that in wild-type mice and continued to rise until 5 months of age. Continuous treatment with halofuginone from 4 weeks to 5 months of age reduced muscle fibrosis in a phosphorylated-Smad3 inhibition-related manner. Halofuginone also enhanced myofiber hypertrophy, reduced the percentage of centrally nucleated myofibers, and increased muscle performance. Together, the data suggest an inhibitory effect of halofuginone on the muscle histopathology at very early stages of dysferlinopathy, and better generation of force and muscle performance. These results offer new opportunities for early pharmaceutical treatment in dysferlinopathies with favorable outcomes at later stages of life.

Crangon crangon: can hydroxyproline be an indicator of changes in the species?

The hydroxyproline content in Crangon crangon tissues from the Gulf of Gdansk (southern Baltic) was determined in males, non-ovigerous females and ovigerous females, depending on the individual body length, the study area (two profiles: Gdynia and Sopot) and the depth of occurrence. Individuals were collected and analyzed from April to August 2008. The research on the migratory species C. crangon indicates that the area and depth of its occurrence do not significantly affect the level of hydroxyproline in the tissues of this animal (p > 0.05). However, certain trends have been observed. Hydroxyproline participates in various life processes of C. crangon and its level in the tissues is significantly correlated with the sex of animals (p < 0.05). In males, hydroxyproline plays a major role in the body growth. Moreover, water temperature significantly affects the hydroxyproline content in males of different body sizes. Ovigerous females use hydroxyproline in the reproductive process. In non-ovigerous females, hydroxyproline participates both in the growth of organisms and in the reproductive period. Hydroxyproline can be an indicator of the collagen level, as well as an important factor in physiological processes.

EFEK SALEP EKSTRAK PINANG TERHADAP LEVEL FIBROBLAST DAN KOLAGEN PADA PROSES PENYEMBUHAN LUKA

ABSTRACT Background: Some plants, such as betel nuts (Areca catechu), is used as traditional antiseptic. Betel nut mash is applied on ulcus wound. Fibroblast and collagen are important factor in wound healing. Research objective : this study is to determine the effect of betel nuts extract on fibroblast and collagen level in full thickness wound healing processs. Methodology: this study used male rats Sprague dawney that randomly divided into 3 groups. Groups I received no treatment, groups II and III received areca catechu extracts with concentration 15% and 30% respectively. There are 12 rats in each groups. The treatment is given every day without wound’s debridement. Incision were made full thickness with diameter size 1,5 cm, on right back skin was made by lidocain anesthesia subcutaneously. Wound area were measured every days. Termination of rats were done in day 7 and day 16 to histopathology assessment with Haematoxylin-Eosin stain for fibroblast and collagen level by semiqualitative score. Results: level of collagen were higher in group that received extract but level of fibroblast were lower than control group in histopathology of day 7th. Conclusion: Extract of betel nut increased level of collagen. Keyword : areca catechu, betel nuts, wound, fibroblast, collagen, histopathology ABSTRAK Latar belakang : Beberapa tanaman digunakan sebagai antiseptik luka, salah satunya biji pinang untuk penyembuhan luka ulkus. Fibroblast dan kolagen merupakan salah satu faktor penting dalam penyembuhan luka. Tujuan : Penelitian ini bertujuan mengetahui efek ekstrak biji pinang terhadap level fibroblast dan kolagen pada proses penyembuhan luka full thickness Metode : Penelitian ini menggunakan tikus galur Sprague dawney jantan berusia 2-3 bulan dan telah mendapatkan persetujuan etik. Setelah aklimatisasi, tikus dibagi secara acak masing-masing 12 ekor dalam 3 kelompok, yaitu kelompok I tanpa perlakuan, kelompok II diberikan salep ekstrak biji pinang 15%, kelompok III diberikan salep ekstrak biji pinang 30%. Luka full thickness dibuat dengan diameter 1,5 cm di daerah punggung belakang bagian kanan dengan anestesi lidokain subkutan. Perlakuan dilakukan setiap hari tanpa debridemen luka. Luas luka diukur setiap hari. Terminasi dilakukan pada hari ke 7 dan 16 untuk pemeriksaan histopatologi jaringan luka dengan pengecatan Haematoxylin-Eosin. Skoring secara semikualitatif untuk menilai fibroblast dan kolagen. Hasil : Pada kelompok ekstrak pinang terdapat peningkatan level kolagen, akan tetapi tidak terjadi peningkatan level fibroblast pada hasil histopatologi luka kulit hari ke-7. Kesimpulan : terjadi peningkatan level kolagen pada pemberian ekstrak biji pinang. Kata kunci : biji pinang, luka, histopatologi, fibroblast, kolagen

Assessment of Anti-Ageing Potential of Consciousness Energy Healing Treated DMEM and HFF-1 Cells using Cellular Proliferation and Collagen Metabolism Assays

Skin health and aging are the complex biological process influenced by several intrinsic (or endogenous) and extrinsic (or exogenous) factors. Various skin-based therapies are currently available to rejuvenate the skin, but they might be related with some side-effects such as scarring. The objective of the present study was to evaluate the effect of Consciousness Energy Healing Treatment (The Trivedi Effect®) on the human foreskin fibroblast (HFF-1) cell line and Dulbecco’s Modified Eagle Medium (DMEM) for skin health parameters like cell proliferation and synthesis of collagen. The rate of cellular proliferation in HFF-1 cells was identified, and the results found that the Biofield Energy Treated DMEM significantly (p ≤ 0.001) increased by 152.38% compared to the negative control group. Additionally, the cell proliferation was also significantly increased by 71.43% in the Biofield Energy Treated cells compared to the negative control group. Similarly, the collagen level was significantly (p ≤ 0.001) increased by 60.42% in the Biofield Energy Treated DMEM compared with the negative control group. Hence, the results exhibited a significant improvement of collagen synthesis and cellular proliferation in the Biofield Energy Treated DMEM for improving skin health. It can be concluded that The Trivedi Effect® - Consciousness Energy Healing Treatment might be a complementary and alternative approach with respect to the skin health, anti-aging in DMEM compared with the HFF-1 cell line. Therefore, the Biofield Energy Treated DMEM could be useful for the development of effective cosmetic products for the prevention and treatment of several skin problems such as erythema, contact dermatitis, skin aging, wrinkles, etc.