Effect of mycophenolate mofetil alleviates carbon tetrachloride-induced liver fibrosis in mice / 中南大学学报(医学版)

OBJECTIVES@#Hepatic fibrosis is a serious pathological consequence of chronic liver disease. Mycophenolate mofetil (MMF) is a commonly used immunosuppressant after organ transplant. However, the relationship between MMF and hepatic fibrosis remains unclear. This study aims to explore the effect of MMF on hepatic fibrosis in mice and the potential mechanism.@*METHODS@#A total of 24 mice (male, 8-week old, C57BL/6) were randomly divided into a control group, a MMF group, a carbon tetrachloride (CCl4) group and a CCl4+MMF group (n=6 in each group). After the mice were sacrificed, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were detected. The liver tissues were taken up for Masson staining and collagen I (COL1) immunohistochemistry. The levels of transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) were detected by Western blotting. Finally, the levels of mRNA for TGF-β1, α-SMA, and COL1 were detected using real-time PCR.@*RESULTS@#Compared with the CCl4 group, the ALT and AST levels were lower (both P<0.05), the degree of liver fibrosis was alleviated, and the deposition of COL1 in the liver was significantly decreased (P<0.01) in the CCl4+MMF group. Compared with the CCl4 group, the protein expression levels of TGF-β1 and α-SMA were significantly decreased (both P<0.05) and the relative expression levels of TGF-β1, α-SMA and COL1 mRNA in the liver were significantly decreased (all P<0.05) in the CCl4+MMF.@*CONCLUSIONS@#MMF could reduce CCl4-induced hepatic fibrosis, which might be related to the inhibition of TGF-β1. This study is expected to provide a target for the treatment of hepatic fibrosis.

Inhibiting collagen I production and tumor cell colonization in the lung via miR-29a-3p loading of exosome-/liposome-based nanovesicles

The lung is one of the most common sites for cancer metastasis. Collagens in the lung provide a permissive microenvironment that supports the colonization and outgrowth of disseminated tumor cells. Therefore, down-regulating the production of collagens may contribute to the inhibition of lung metastasis. It has been suggested that miR-29 exhibits effective anti-fibrotic activity by negatively regulating the expression of collagens. Indeed, our clinical lung tumor data shows that miR-29a-3p expression negatively correlates with collagen I expression in lung tumors and positively correlates with patients' outcomes. However, suitable carriers need to be selected to deliver this therapeutic miRNA to the lungs. In this study, we found that the chemotherapy drug cisplatin facilitated miR-29a-3p accumulation in the exosomes of lung tumor cells, and this type of exosomes exhibited a specific lung-targeting effect and promising collagen down-regulation. To scale up the preparation and simplify the delivery system, we designed a lung-targeting liposomal nanovesicle (by adjusting the molar ratio of DOTAP/cholesterol-miRNAs to 4:1) to carry miR-29a-3p and mimic the exosomes. This liposomal nanovesicle delivery system significantly down-regulated collagen I secretion by lung fibroblasts in vivo, thus alleviating the establishment of a pro-metastatic environment for circulating lung tumor cells.

Interventional effect of asiaticosdide on rats exposed to silica dust / 中华劳动卫生职业病杂志

Objective: To investigate the effect of asiaticoside for fibrosis in lung tissues of rats exposed to silica and to explore its possible mechanism. Methods: 144 SD male rats were randomly divided into control group, model group, positive drug control group, asiaticoside high-dose group, medium-dose group and low-dose group, each group included 24 rats. Rats in the control group were perfused with 1.0 ml of normal saline, and the other groups were given 1.0 ml 50 mg/ml SiO(2) suspension. Gavage of herbal was given from the next day after model establishment, once a day. Rats in the positive drug control group were administration with 30 mg/kg tetrandrine and rats in the low-dose group, medium-dose group and high-dose group were given 20 mg/kg, 40 mg/kg and 60 mg/kg asiaticoside for fibrosis respectively. Rats in the control group and the model group were given 0.9% normal saline. The rats were sacrificed in on the 14th, 28th and 56th day after intragastric administration and collect the lung tissues to detect the content of hydroxyproline, TGF-β(1) and IL-18, observe the pathological changes of the lung tissues by HE and Masson staining and determine the expressions of Col-I, a-SMA, TGF-β in lung tissues by Western Blot. Results: On the 14th day, 28th day and 56th day after model establishment, the lung tissues of rats in the model group showed obvious inflammatory response and accumulation of collagen fibers, and the degree of inflammation and fibrosis increased with time. The intervention of asiaticoside could effectively inhibit the pathological changes of lung tissues. The contents of hydroxyproline, IL-18 and TGF-β1 in lung tissues of model group were higher than those in the control group (P<0.05) , while the level of hydroxyproline, IL-18 and TGF-β1 in asiaticoside groups were significantly decreased, and the difference was statistically signicant (P<0.05) . Compared with the control group, the expression levels of Col-I, TGF-β1and α-SMA in lung tissue of model group were increased (P<0.05) , while the expression level of Col-I, TGF-β1 and α-SMA were decreased after the intervention of asiaticoside, and the difference was statistically signicant (P<0.05) . Conclusion: Asiaticoside can inhibit the increase of Col-I, TGF-β1 and α-SMA content in the SiO(2)-induced lung tissues of rats, reduce the release of TGF-β1 and IL-18 inflammatory factors in lung tissue, and then inhibit the synthesis and deposition of extracellular matrix in rat lung tissue, and improve silicosis fibrosis.

CircularRNA CDR1as promotes osteogenic differentiation and angiogenesis related genes expression in mouse bone marrow mesenchymal stem cells / 口腔疾病防治

Objective@# To investigate the effects of over expression and low expression of antisense transcripts of circular RNA cerebellar degeneration associated protein 1 (CDR1as) in Balb/C mouse bone marrow mesenchymal stem cells (BMSCs) on factors related to osteogenesis and angiogenesis.@*Methods@#BMSCs were cultured and identified in vitro. The lentiviral (LV) vector containing the overexpressed and silenced circRNA CDR1as genes and the control lentivirus were respectively transfected into mouse BMSCs, and stable cell lines were screened. The cells were divided into the circRNACDR1as over expression group and the over expression control group, and the CircRNACDR1as low expression group and the low expression control group. The components were stained with Alizarin Red S and alkaline phosphatase after 14 and 21 days of osteoinduction; qRT-PCR was used to detect the target genes circRNA CDR1as, osteogenic differentiation markers alkaline phosphatase (ALP), runt- related transcription factor 2 (RUNX2), osteocalcin (OCN), osteopontin (OPN), osterix(Osx), collagen I (COL-1), and the mRNA expression levels of vascular endothelial grown factor (VEGF) and angiogenin-1 (Ang-1). @*Results@# The results of alizarin red staining and alkaline phosphatase staining showed that the extracellular matrix calcium precipitation and ALP staining area of the over expression experimental group was greater than its control group, and those of the low expression experimental group was less than its control group. As the number of days of osteogenic induction increased, the calcium precipitation and ALP staining in each group also increased. RT-PCR results showed that the mRNA expression levels of circRNA CDR1as, ALP, RUNX2, OCN, OPN, OSX, COL-1, VEGF and Ang-1 in the over expression experimental group BMSCs were significantly increased (P<0.001). In the low expression experimental group, the mRNA expression levels of circRNA CDR1as, ALP, RUNX2, OCN, OPN, OSX, COL-1, VEGF and Ang-1 in BMSCs were significantly reduced (P<0.001). @*Conclusion@# Over expression of the circRNA CDR1as gene promotes the osteogenic differentiation and angiogenesis of BMSCs. Low expression of the circRNA CDR1as gene inhibits the osteogenic differentiation and angiogenesis of BMSCs.

Effect of Tetrandrine on Col-I and FN in TGF-β1-induced MRC-5 Cells / 中国实验方剂学杂志

Objective:To investigate the effect of tetrandrine on transforming growth factor-β1（TGF-β1）stimulated MRC-5 cells. Method:Different concentrations of TGF-β1 (0, 2.5, 5, 10, 20, 40 μg·L-1) were applied to MRC-5 cells. Proliferation toxicity of TGF-β1 to MRC-5 was detected by cell counting kit-8 (CCK-8) method. Detection of alpha smooth muscle actin (α-SMA) and Vimentin's expression levels in MRC-5 by Western blot. Detection of changes of collagen I(Col-I) and fibronectin (FN)'s expression levels in MRC-5 supernatants by enzyme linked immunosorbent assay(ELISA) kit. And the appropriate concentration of TGF-β1 activated MRC-5 cells was screened. The appropriate concentration of TGF-β1 and different concentrations of Tet (0, 2.5, 5, 10, 20, 40 μmol·L-1) were applied to MRC-5 cells, and CCK-8 method was used to screen safe concentration again. Western blot was used to detect changes in α-SMA and Vimentin expression levels in MRC-5 cells, and ELISA method to detect changes in Col-I and FN in MRC-5 cell supernatant. Result:Compared with the blank group, 20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 24 hours (P<0.05), and 10，20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 48 h (P<0.05).When Tet is added for 24 h, the half inhibitory concentration (IC50) value was 14.07 μmol·L-1, and when cultured for 48 h, the IC50 value was 7.51 μmol·L-1. Compared with the blank group, the relative contents of α-SMA, FN and Col-I in the 5 μg·L-1 of TGF-β1 group were obviously increased (P<0.05), and the relative contents of Vimentin were significantly increased (P<0.01), and the relative contents of FN and Col-I, α-SMA and Vimentin in 10 μg·L-1 group were significantly increased (P<0.01). 10 μg·L-1 of TGF-β1 was co-cultured with Tet at different concentrations. Compared with the TGF-β1 group, the relative levels of α-SMA, Vimentin and FN in the 5 μmol·L-1 of Tet group were significantly reduced (P<0.01), and the relative levels of Col-I were obviously reduced (P<0.05). In the Tet 10 μmol·L-1 group, the relative contents of the α-SMA, Vimentin, FN and Col-I were significantly reduced (P<0.01). Conclusion:TGF-β1 can increase the levels of Col-I, FN and other extracellular matrices in MRC-5 cells, and Tet can effectively inhibit the occurrence of this change. It is suggested that Tet may inhibit secreting extracellular matrix of fibroblasts in the formation of pulmonary fibrosis.

Effect of mir-152 on proliferation of myocardial fibroblasts in diabetic cardiomyopathy / 中国药理学通报

Aim To explore the effect of miR-152 on proliferation of cardiac fibroblasts ( CFS) in diabetic cardiomyopathy. Methods Diabetic cardiomyopathy model was established in SD rats by STZ injection, and CFS proliferation model was established by high glucose (33. 3 mmol • L ~1). HE and Masson staining were performed in paraformaldehyde fixed myocardium of rats. Western blot determined a-SMA and collagen I protein expression. qPCK detected gene expression of miR-152. MTT assay analyzed the proliferation of cells. Results HE and Masson staining showed the higher level of myocardial collagen in diabetic cardiomyopathy model. Furthermore, the myocardial myo-cytes lined up in disorder. Western blot showed that the expressions of a-SMA and collagen I were up-regulated in the diabetes mellitus ( DM ) group, while the expression of miR-152 was down-regulated. The result of the in vitro experiment showed that a-SMA and collagen I expressions were down-regulated after trans-fected miR-152 mimics. The proliferation of CFS was also down-regulated after transfected miR-152 mimics. Conclusions miR-152 plays an important role in the proliferation of CFS and may ameliorate diabetic cardiomyopathy.

Effects of total C-21 steroidal glucosides from Cynanchum auriculatum on renal fibrosis in rats and mechanisms involved / 中草药

Objective: To investigate the effects of C-21 steroidal glucosides from Cynanchum auriculatum on renal fibrosis in rats caused by unilateral ureteral ligation and explore its mechanisms. Methods: A total of 15 rats were randomly chosen as sham operation group (Sham), while the remaining rats underwent unilateral ureteral ligation. The rats after the operation were randomly divided into four groups, namely the Sham operation group, the model group, the positive control group (20 mg/kg), C. auriculatum high group (400 mg/kg), C. auriculatum low group (200 mg/kg), 15 per group, and the rats in each group were administrated with corresponding drugs. After the intervention of 28 d, all the rats were sacrificed and the kidneys were then removed. The content of hydroxyproline was measured. HE and Masson staining were conducted to assess kidney pathological changes and renal fibrosis. The protein expression of TGF-β1, α-SMA, and E-cadherin were tested with immunohistochemistry and Western blotting. The mRNA expression of collagen-I and collagen-III was evaluated using qRT-PCR. Results: Compared with the model group, C-21 steroid glycosides significantly alleviated the kidney pathological injury and renal fibrosis, and reduced fibrous tissue and collagen proliferation. C-21 steroid glucosides from C. auriculatum can significantly reduce the kidney/body weight ratio and the content of hydroxyproline in kidney tissue (P < 0.05, 0.01), and it showed in a dose-dependent manner. Compared with the model group, the expression of α-SMA and TGF-β1 was decreased significantly, and the expression of E-cadherin was increased significantly (P < 0.05, 0.01) in C. auriculatum group. Moreover, compared with the model group, C-21 steroidal glucosides significantly down-regulated the mRNA expression of collagen-I and collagen-III mRNA (P < 0.01). Conclusion: C-21 steroid glycosides from C. auriculatum can effectively attenuate renal fibrosis in rats with unilateral ureteral ligation and its underlying mechanisms may be related to inhibiting the over-expression of collagen-I and collagen-III, down-regulating the expression of α-SMA and TGF-β1, and up-regulating the expression of E-cadherin. By regulating the tubular epithelial-mesenchymal transition, it exerts its anti-fibrotic effect.

The role of transforming growth factor β1 and collagen in vulvar lichen sclerosis / 医学研究生学报

ObjectiveTo investigate the expression of transforming growth factor β1 (TGF-β1) and collagen I, III (Col I, III) in vulvar lichen sclerosis (VLS) and their role in VLS.MethodsThe specimens of30 VLS tissues (15 in early stage and 15 in progressive stage), and 15 vulvar normal skin tissues were selected by biopsy or surgical excision from 2016 to 2018 in our hospital. The expression of Vimentin was detected by immunohistochemistry. The expression of TGF-β1, TGF-β2, Col I and Col III mRNA was detected through RT-PCR. Simultaneously, the fibroblasts were visualized in diseased tissues by labeling Vimentin.ResultsThe expression of Vimentin in VLS was increased significantly (P0.05). Col I mRNA was up-regulated in VLS, obviously in early stage. Meanwhile, Col III mRNA down-regulated gradually from the early to the progressive stage of VLS. Therefore, the Col I/III ratio increased gradually.ConclusionThe increase of fibroblasts and TGF-β1 in dermis of VLS promotes the synthesis of Col I and reduces the content of Col III, which may be one of the factors leading to the decrease of skin elasticity in VLS.

Effect of diabetes on CCl4-induced rats liver fibrosis / 中国药理学通报

Aim To observe the effect of diabetes on carbon tetrachloride (CCl4)-induced rats chemical liv-er injury and liver fibrosis by establishing diabetes mer-ged with liver fibrosis rat model (double model). Methods High fat feeding combined with streptozoto-cin (STZ) was used to induce diabetes rat model,and liver fibrosis rat model was induced by CCl4. HE stai-ning was used to observe the rat liver pathological changes, and Western blot and q-PCR were used to detect liver fibrosis related factor genes α-SMA and Collagen Ⅰ expression. Results Compared with con-trol group, diabetes group, liver fibrosis group and double model group all had different levels of liver damage,especially double model group. Rat liver tis-sues of α-SMA and CollagenⅠexpression from differ-ent model groups also increased, especially those from double model group, and significant differences were detected compared to diabetes and liver fibrosis group. Conclusions Diabetes can cause liver damage and in-crease the occurrence and development of CCl4-in-duced rats liver fibrosis, and the mechanism may be related to the formation and/or degradation of extracel-lular matrix.

Effects of inflammatory microenvironment mediated by macrophage on the proliferation and osteogenic differentiation of periodontal ligament cells / 口腔疾病防治

Objective@#The present study investigated the effects of the inflammatory microenvironment mediated by macrophages on the proliferation and osteogenic differentiation of periodontal ligament cells (PDLCs).@*Methods@#Conditioned medium containing inflammatory factors was collected following macrophage activation with 1 μg/mL lipopolysaccharide (LPS). PDLCs were isolated from healthy teeth and cultured in conditioned medium (LPS-CM group) or normal medium (control group), and the proliferation of PDLCs was detected using the MTT assay. The cells were cocultured with an osteogenic inducer for 3 d and 7 d, and the alkaline phosphatase (ALP) activity of PDLCs was detected using an ALP kit. The mRNA expression levels of runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and collagen I (COL-I) were detected using real-time PCR, and the protein levels of RUNX2, OCN, and COL-I were detected using Western blotting. Mineralization nodules were observed using Alizarin red staining after osteoinduction for 14 d. @*Results@#The OD value of PDLCs in the LPS-CM group was lower than that in the control group (P < 0.05). The mRNA levels of RUNX2, OCN, and COL-I in the LPS-CM group were lower than those in the control group (P < 0.05). In addition to the OCN 3 d group (t = 2.75, P = 0.056), the protein expression of RUNX2, OCN, and COL-I in the LPS-CM group was lower than that in the control group (P < 0.05). However, the ALP activity of the LPS-CM group was higher than that of the control group, which was 1.58-fold greater (t = 5.91, P = 0.030) at 3 d and 1.29-fold greater (t = 6.01, P = 0.046) at 7 d. The number of calcified nodules in the LPS-CM group was significantly less than that in the control group (t = 8.63, P = 0.048). @*Conclusion@# The inflammatory microenvironment mediated by macrophages may inhibit the proliferation and osteogenic differentiation of PDLCs.

The effect of exogenous VEGF on bone activity of rabbit heterotopic allograft decalcified bone / 中华显微外科杂志

Objective To investigate the effect of exogenous vascular endothelial growth factor (VEGF) on bone activity of rabbit heterotopic allograft decalcified bone.Methods 140 adult healthy China white rabbits were selected,no limitation with sex,20 rabbits as the donor preparation of allogenic decalcified bone,according to the random number table,the rest was divided into the experimental group (allograft decalcified bone ± VEGF) and the control group (Allograft decalcified bone),each group contained 60 rabbits.For the experimental group,the prepared 1.5 cm long homologous decalcified tibia was placed in rabbit right thigh of rectus femoris and vastus medialis muscle gap near by saphenous artery,and fixed on the femur with two 0.8 mm Kirschner wire.In the vicinity of the skin,implanted an osmotic pump which contain the VEGF solution 200 μl with concentration was 0.5 μg/ml.In the control group,implanted the isometric allograft decalcified bone in rabbit right thigh corresponding parts with the same method.Each group respectively at 0,2,4,6,8,10 weeks to death 10 white rabbits,By specimen observation,HE dyeing observation and detection of type Ⅰ glue protein fluorescence intensity,Analysis the bone activation degree of two groups of bone allograft decalcified.Results Experimental allograft decalcified bone gradually wrapped by connective tissue membrane,its surface appear different size of the pits and gradually increased and become deep,while the control group pits relatively little and shallow.In the experimental group and control group,the fluorescence intensity of type Ⅰ collagen reached its peak respectively at 8 weeks (47.57 ±3.50) and 10 weeks (45.07±6.02),with no statistically significant (P ＞ 0.05).Conclusion Rabbit allograft decalcified bone implanted in the muscle clearance with abundant blood supply can be transformed into activated bone after 10 weeks,and after applying exogenous VEGF,allograft decalcified bone can be transformed into activated bone after 8 weeks,the bone activation process obviously speed up.The reaults confirmed the exogenous VEGF can obviously promote the ectopic rabbit bone allograft decalcified bone activation process.

Effects of CUEDC2 on inflammation and renal interstitial fibrosis in obstructive nephropathy / 实用医学杂志

Objective To study the effects of CUEDC2 on renal interstitial fibrosis and inflammation response in rats with unilateral ureteral obstruction (UUO). Methods 30 Balb/c rats were randomly distributed into sham operation group(sham-vector),uuo operation group(uuo-vector) and CUEDC2 treatment group after uuo (uuo-cuedc2). Hematoxylin-eosin and Masson staining were used to measure renal pathology; Inflammation factors were quantified by ELISA; Immunohistochemistry was performed to measure the expression of CUEDC2;Protein expression of CUEDC2, Fibronectin, E-cadherin, Collagen I were detected by Western Blot. Results At 7 and 14d after operation, the area of interstitial fibrosis and expression of ICAM1,MCP1,IL1,IL8, Fibronectin and Collagen I in uuo-cuedc2 showed a marked decrease when compared to uuo-vector (p?0.05),the level of E-cadherin was significantly increased (P < 0.05). Conclusion CUEDC2 can inhibit renal interstitial fibrosis and decrease the expression of inflammation factors and Collagen deposition.

The Efficacy and Safety of Collagen-I and Hypoxic Conditions in Urine-Derived Stem Cell Ex Vivo Culture

Upper urinary tract-derived urine stem cells (USCs) are considered a valuable mesenchymal stem cell source for autologous cell therapy. However, the reported culture condition for USCs is not appropriate for large-quantity production, because cells can show limited replicativity, senescence, and undesirable differentiation during cultivation. These drawbacks led us to reconstitute a culture condition that mimics the natural stem cell niche. We selected extracellular matrix protein and oxygen tension to optimize the ex vivo expansion of USCs, and compared cell adhesion, proliferation, gene expression, chromosomal stability, differentiation capacity, immunity and safety. Culture on collagen type I (ColI) supported highly enhanced USC proliferation and retention of stem cell properties. In the oxygen tension analysis (with ColI), 5% O₂ hypoxia showed a higher cell proliferation rate, a greater proportion of cells in the S phase of the cell cycle, and normal stem cell properties compared to those observed in cells cultured under 20% O₂ normoxia. The established reconstituted condition (ColI/hypoxia, USCs(recon)) was compared to the control condition. The expanded USCs(recon) showed highly increased cell proliferation and colony forming ability, maintained transcription factors, chromosomal stability, and multi-lineage differentiation capacity (neuron, osteoblast, and adipocyte) compared to the control. In addition, USCs(recon) retained their immune-privileged potential and non-tumorigenicity with in vivo testing at week 8. Therefore, reconstituted condition allows for expanded uUSC cell preparations that are safe and useful for application in stem cell therapy.

Collagen I-induced dendritic cells activation is regulated by TNF-α production through down-regulation of IRF4.

Previously we have shown that collagen I enhances the maturation and function of dendritic cells (DCs). Inflammatory mediators such as tumour necrosis factor (TNF)-α, interleukin (IL)-1β and lipopolysaccharide (LPS) are also known to activate DCs. Here we investigated the involvement of TNF-α on the collagen I-induced DCs activation. TNF-a neutralization inhibited collagen I-induced IL-12 secretions by DCs. Additionally, we observed suppression of collagen I-induced costimulatory molecules expression along with down-regulation of genes involved in DCs activation pathway. Furthermore, TNF-α inhibition upon collagen Istimulation up-regulated the expression of interferon regulatory transcription factor IRF4, when compared to collagen I only treated cells. Collectively, our data demonstrate that collagen I induce TNF-α production, which is crucial for the activation and function of DCs, through down-regulation of IRF4, and implicates the importance in development of anti- TNF-α therapeutics for several inflammatory diseases.

Platelet-derived growth factor mediates interleukin-13-induced collagen I production in mouse airway fibroblasts.

Interleukin-13 (IL-13) is associated with the production of collagen in airway remodelling of asthma. Yet, the molecular mechanisms underlying IL-13 induction of collagen remain unclear; the aim of this study is to address this issue. IL-13 dose- and time-dependently-induced collagen I production in primary cultured airway fibroblasts; this was accompanied with the STAT6 phosphorylation, and pre-treatment of cells with JAK inhibitor suppressed IL-13- induced collagen I production. Further study indicated that IL-13 stimulated JAK/STAT6-dependent PDGF production and subsequent ERK1/2 MAPK activation in airway fibroblasts, and the presence of either PDGF receptor blocker or MEK inhibitor partially suppressed IL-13-induced collagen I production. Taken together, our study suggests that activation of JAK/STAT6 signal pathway and subsequent PDGF generation and resultant ERK1/2 MAPK activation mediated IL-13-induced collagen I production in airway fibroblasts.

RETRACTED: Effect of phentolamine on myocardial extracellular matrix of cardiac remodeling in rats

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal).This article has been retracted at the request of the journal Editorial Office.The authors have plagiarized part of a paper that had already appeared in Chinese Journal of Arteriosclerosis 2014, 4, 362–366. article id: 1007–3949 (2014) 22-04-0362-05. One of the conditions of submission of a paper for publication is that authors declare explicitly that their work is original and has not appeared in a publication elsewhere. Re-use of any data should be appropriately cited. As such this article represents an abuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of the journal that this was not detected during the submission process.

A study on prothrombogenic and antithrombogenic biomarkers in deep vein thrombosis following meta-epiphyseal cancellous bone traumatization in major orthopedic surgeries.

Background: This study was aimed to show differences in the mean values of Collagen I (Col I)/Nitric Oxide (NO), Collagen IV (Col IV)/NO, Tissue Factor (TF)/NO, and P-selectin (P-sel)/NO ratios between patients with DVT and those without DVT, following hip and knee meta-epiphyseal cancellous bone traumatization in major orthopedic surgeries. Methods: This is an observational prospective cohort study on 69 patients aged > 50 years, who had orthopedic surgery without thromboprophylaxis. Examination of serum Col I, Col IV, TF, P-sel, and NO biomarker levels were performed three times, i.e. before surgery, 72 hours and 144 hours after surgery. We looked for the differences in mean levels of biomarkers, and mean ratio values of the prothrombogenic/antithrombogenic (Col I/NO, Col IV/NO, TF/NO, P-sel/NO) at 72 hours and 144 hours post surgery between patients with DVT and those without. DVT events, which were confirmed at 144 hours post surgery by venography (with the exception of 8 cases where color Doppler ultrasound was done due to contrast usage contraindications). Results: DVTs were identified in 18 patients (26.1%). There were significant differences of mean levels in pre-surgical Col IV (p = 0.022) and 72 hours NO (p = 0.014) between patients with and without DVT. In addition, between the same two patient groups, significant differences were found in the mean values of the prothrombogenic/antithrombogenic ratios, i.e. Col IV/NO, TF/NO, and P-sel/NO at 72 hours post-surgery (p = 0.007, p = 0.028, and p = 0.049 respectively), with lower median values that were found in subjects with DVT. At 144-hours post surgery, the only significant ratio difference between the two groups was the mean values of Col IV/NO ratio (p = 0.014) with the median values that were higher than the median values at 72-hours post surgery . Conclusion: The incidence of DVT following traumatization of the meta-epiphyseal cancellous bone after major orthopedic surgeries in hip and knee is influenced by the balance of prothrombogenic and antithrombogenic factors as shown by the significant differences in Col IV/NO, TF/NO and P-sel/NO ratios at 72-hours and Col IV/NO ratio at 144 hours after surgery between DVT positive and DVT negative patients.

Boron as testicular toxicant in mice (Mus domesticus) / El boro como tóxico testicular en ratón (Mus domesticus)

It has been reported that boron causes changes in various systems, including the male reproductive system. Residents in some towns in northern Chile were consuming a few years ago in the drinking water 20 times more than the amount established as permissible limit by WHO. This study evaluates the effects in an animal model of high intake of boron on the testis. Boron was administered in the drinking water. Twenty male mice (Mus domesticus), sexually mature, were used, divided into 2 groups: the experimental group was given Boron at a dose of 12 mg/L, and the control group 0.6 mg/L, for 42 days. Sections of testis were obtained for: HE staining (Morphometry and Histopathology), Immunohistochemistry (Cox-2), Mallory and Picrosirious stain (evaluation of tunica albuginea). The results indicate that ingestion of a dose of 12 mg Boron/L produces vacuolization, tubular epithelial desquamation and tamponade. Morphometry revealed decreased tubular diameter and epithelial height and lumen diameter and increased interstitial area in the exposed group. Immunodetection of COX-2 was positive in high percentage of tubules in the intoxicated group. The tunica albuginea was thinner, with decreased percentage of type I collagen fibers and an increase in the percentage of type III collagen fibers in animals exposed to boron in contrast to the control group. Exposure to critical levels of boron produces severe histopathological changes in the testis, altering morphometric parameters and causing overexpression of Cox-2. Finally, evaluation of collagen fibers suggests that Boron produced a degradation of the collagen of the tunica albuginea, causing a decrease in the thickness of it and altering the percentage ratio collagen I/collagen III, a process called collagenolysis...

Se ha reportado que el Boro provoca alteraciones en diversos sistemas, incluido el sistema reproductor masculino. Los habitantes de algunas localidades del norte de Chile estuvieron hasta hace algunos años consumiendo en el agua de bebida 20 veces más que la cantidad establecida como límite permisible por la OMS. El presente estudio evalúa en modelo animal los efectos del alto consumo de Boro sobre el testículo, administrado en el agua de bebida. Se utilizaron veinte ratones (Mus musculus) machos, sexualmente maduros, divididos en 2 grupos; al grupo experimental se administró Boro a una dosis de 12 mg/L, y al grupo control 0.6 mg/L, durante 42 días. Posteriormente se obtuvieron secciones de testículo para: Tinción H-E (Morfometría e Histopatología); Inmunohistoquímica (Cox-2); Tinción de Mallory y Picrosirious Red (evaluación de túnica albugínea). Los resultados indican que la ingestión de Boro a dosis de 12 mg/L produce vacuolización, descamación epitelial y taponamiento tubular. La morfometría revela disminución en el diámetro tubular y altura epitelial, así como aumento del diámetro luminal y del área intersticial en el grupo expuesto. La inmunodetección de COX-2 resultó positiva en gran porcentaje de túbulos en el grupo intoxicado. La túnica albugínea, demostró menor grosor, así como una disminución en el porcentaje de fibras colágenas tipo I y un aumento en el porcentaje de fibras colágenas tipo III en los animales expuestos a Boro en contraste con el grupo control. La exposición a niveles críticos de Boro genera alteraciones histopatológicas severas en el testículo, alterando parámetros morfométricos y provocando la sobreexpresión de COX-2. Finalmente, la evaluación de fibras colágenas sugiere que el Boro produjo una degradación del colágeno de la túnica albugínea, provocando una disminución en el espesor de ésta y alterando la relación porcentual Colágeno I/Colágeno III, proceso llamado colagenólisis...

Possible molecular mechanism of promotion of repair of acute achilles tendon rupture by low intensity-pulsed ultrasound treatment in a rat model / Posible mecanismo molecular de promoción de la ruptura aguda del tendón de aquiles por el tratamiento de ultrasonidos pulsados de baja intensidad en un modelo de rata

OBJECTIVE: This study investigated the effect of Low Intensity-pulsed Ultrasound (LIPUS) on the repair process of ruptured Achilles tendon using a rat model and also examined the regulation of a biological molecule that may contribute to this in vitro and in vitro. METHODS: To investigate the effect of LIPUS and its biological mechanism ofpromoting Achilles tendon repair after acute injury, ninety-eight male Sprague-Dawley (SD) rats (mean body weight, 258 ±9.8 g) aged 12 weeks were used in this study. To create the model, the Achilles tendon attachment site and musculotendinous junction were ruptured under direct vision. The leg on one side was exposed to LIPUS (frequency at 1.5 MHz, the repetition cycle at 1.0 kHz, the burst width at 200 msec and the power output at 45 mW/cm2), for 20 minutes daily with a 0.7 mm diameter probe. Results:Low Intensity-pulsed Ultrasound treatment accelerated the repair of the Achilles tendon compared to the untreated group, judged by electron microscopy. Both cyclo-oxygenase (COX)-2* and EP4* expressions were over-expressed in the LIPUS treated group in the inflammatory period, and TGFJ31* expression was markedly induced in LIPUS treated groups followed by collagen I* and III* expression in the repair and reconstitution process. CONCLUSION: These findings suggest that LIPUS is potentially able to accelerate the repair of acute ruptured Achilles tendon in several ways: by exaggerating inflammation by inducing COX-2 and EP4 and reconstituting tissue by inducing TGFJ31 followed by collagen I and III. (*: p < 0.05, **: 0.001).

OBJETIVO: Este estudio estuvo encaminado a investigar el efecto de los ultrasonidos pulsados de baja intensidad (LIPUS) sobre el proceso de reparación del tendón de Aquiles tras una ruptura, usando un modelo de rata. Asimismo, se examinó la regulación de una molécula biológica que puede contribuir a este proceso in vitro e in vitro. MÉTODOS: Con el fin de investigar el efecto de LIPUS y el mecanismo biológico por el cual este efecto promueve la reparación del tendón de Aquiles tras una lesión aguda, noventa y ocho ratas machos Sprague-Dawley (SD) (peso corporal promedio, 258 ± 9.8 g) de 12 semanas de edad fueron usadas en este estudio. Para crear el modelo, el sitio de ligazón microbiológica del tendón de Aquiles y la unión músculo-tendinosa fueron desgarrados bajo visión directa. La pierna de un lado fue expuesta a LIPUS (frecuencia de 1.5 MHz, ciclo de repetición de 1.0 kHz, ancho de ruptura de 200 msec, y potencia de salida de 45 mW/cm2), por 20 minutos diariamente con una sonda de 0.7 mm diámetro. RESULTADOS: El tratamiento de ultrasonidos pulsados de baja intensidad aceleró la reparación del tendón de Aquiles, en comparación con el grupo no tratado, según se apreció mediante el microscopio electrónico. Tanto la ciclo-oxygenasa (COX)-2* como las expresiones EP4* estuvieron sobe-expresadas en el grupo tratado con LIPUS en el periodo inflamatorio, y la expresión TGFfi1* fue marcadamente inducida en los grupos tratados con LIPUS seguidos por la expresión de colágeno I* y III* en el proceso de reparación y reconstitución. CONCLUSIÓN: Estos resultados sugieren que LIPUS puede potencialmente acelerar la reparación del tendón de Aquiles luego de un desgarramiento, de varias maneras: exagerando la inflamación mediante inducción de COX-2 y EP4 y reconstituyendo el tejido induciendo TGFfil seguido por colágeno I y III. (*: p < 0.05, **: 0.001).

Effect of gallium-arsenide laser, gallium-aluminum-arsenide laser and healing ointment on cutaneous wound healing in Wistar rats

This study determined the effects of gallium-aluminum-arsenide laser (GaAlAs), gallium-arsenide laser (GaAs) and Dersani® healing ointment on skin wounds in Wistar rats. The parameters analyzed were: type I and III collagen fiber concentrations as well as the rate of wound closure. Five wounds, 12 mm in diameter, were made on the animals’ backs. The depth of the surgical incision was controlled by removing the epithelial tissue until the dorsal muscular fascia was exposed. The animals were anesthetized with ketamine and xylazine via intraperitoneal injection. The rats were randomly divided into five groups of 6 animals each, according to the treatment received. Group 1 (L4): GaAs laser (4 J/cm²); group 2 (L30): GaAlAs laser (30 J/cm²); group 3 (L60): GaAlAs laser (60 J/cm²); group 4 (D): Dersani® ointment; group 5 (control): 0.9 percent saline. The applications were made daily over a period of 20 days. Tissue fragments were stained with picrosirius to distinguish type I collagen from type III collagen. The collagen fibers were photo-documented and analyzed using the Quantum software based on the primary color spectrum (red, yellow and blue). Significant results for wound closing rate were obtained for group 1 (L4), 7.37 mm/day. The highest concentration of type III collagen fibers was observed in group 2 (L30; 37.80 ± 7.10 percent), which differed from control (29.86 ± 5.15 percent) on the 20th day of treatment. The type I collagen fibers of group 1 (L4; 2.67 ± 2.23 percent) and group 2 (L30; 2.87 ± 2.40 percent) differed significantly from control (1.77 ± 2.97 percent) on the 20th day of the experiment.