PCNA-positive Cells Research Articles

Effect of spirulina on corneal epithelial wound healing in zebrafish

This study investigated the regenerative and healing effect of spirulina on corneal epithelial injury by acid burn in zebrafish. Corneal epithelium of adult zebrafish was evaluated at 0, 12, 24, 48 h after acid burn injury with or without immersion with 0.02 and 0.04 mg/mL of spirulina water extract. The pathological changes of corneal epithelium were assessed by light microscopy. Gene and protein expressions of proliferating cell nuclear antigen (PCNA), matrix metalloproteinase (MMP) 9 and MMP13 were determined by immunohistochemistry or real-time PCR. Corneal epithelium was denuded totally after acid burn and gradually regenerated. PCNA-positive cells significantly increased in spirulina treated group. MMP9 and MMP13 mRNA transcripts were significantly decreased in spirulina treated groups. In conclusion, spirulina has regenerative and healing properties by increased keratocyte proliferation, and inhibited extracellular matrix degradation during the regeneration process of corneal epithelial injury by acid burn in zebrafish.

Effects of Oxytetracycline and Gentamicin Therapeutic Doses on Hematological, Biochemical and Hematopoietic Parameters in Cyprinus carpio Juveniles.

Hematological, biochemical and hematopoietic effects of therapeutic doses of two antibiotics, oxytetracycline (OTC) and gentamicin (GEN), in clinically healthy common carp juveniles were studied. The fish were divided into four groups: controls 1 and 2 (untreated or injected with 0.6% NaCl solution), and two groups treated with antibiotics (orally with 75 mg/kg OTC four times every two days or injected with a single dose (4 mg/kg) of GEN dissolved in 0.6% NaCl). Blood and head kidneys were sampled from all fish 3 days post-treatments for hematological, biochemical and hematopoietic tissue analyses. No major alterations in the values of hematological and serum biochemical parameters occurred following administration of OTC or GEN. Glucose concentrations were significantly lower in both groups of fish subjected to injections (Control 2 and GEN), while the oxidative metabolic activity of phagocytes increased in the antibiotic-treated groups (significantly in OTC). More alterations were observed in hematopoietic tissue. Immunocytochemical analysis revealed that G caused a significant increase in the rate of cell proliferation (PCNA-positive cells) and an increase in the frequency of apoptotic cells (caspase-positive). The frequency of lymphoid lineage decreased, which was related to a decrease in the abundance of mature lymphocytes in GEN-treated fish. Percentages of neutrophilic lineage were significantly elevated in OTC and GEN groups compared to controls. The obtained results showed no considerable hematotoxicity or hepatotoxicity of therapeutic doses of OTC and GEN to carp.

Histological and Biochemical Changes in Adult Male Rat Liver after Spinal Cord Injury with Evaluation of the Role of Granulocyte-Colony Stimulating Factor

ABSTRACT Spinal cord injury (SCI) is a devastating disease leading to motor disability. Metabolic dysfunction is another complication of SCI. Thus, we aimed to study the effect of SCI on the histological and biochemical structure of the liver in adult male rats and to delineate the role of post-injury administration of G-CSF. Thirty adult male Sprague-Dawley rats were assigned into three groups: Group I; control (18 rats subdivided equally into three subgroups), and 12 rats underwent SCI and were divided into an SCI group II and G-SCF-treated group III. Twenty-one days post-injury, liver sections were processed for light and electron microscopic examinations and immunohistochemical staining for PCNA and CD68 antibodies. The biochemical assay was carried out for detection of serum levels of ALT, AST, total proteins, albumin, total cholesterol, triglycerides, HDL-c, GSH and MDA. Liver tissue levels of GPx and MDA as well as semiquantitative RT-PCR analysis of hepatic cytokine expression were also conducted. In the SCI group, results showed liver tissue damage in the form of lipid infiltration, blood vessel congestion, vacuolated cells with apoptotic nuclei and increased collagen deposition. Increased CD68-positive macrophages and a decreased number of PCNA-positive cells was detected. Moreover, liver enzymes, total cholesterol and triglycerides were increased while serum albumin, total proteins and HDL-c were decreased in the SCI group. Oxidative stress and increased expression of inflammatory cytokines were detected. Administration of G-CSF induced significant liver improvement with retained liver function by anti-inflammatory, immune-modulatory and antioxidant mechanisms.

Low-polar extract from seed of Cleistocalyx nervosum var. paniala modulates initiation and promotion stages of chemically-induced carcinogenesis in rats

BackgroundCleistocalyx nervosum var. paniala is a local fruit mainly cultivated in the north of Thailand. Our previous study has reported that the methanol extract of C. nervosum seed presented antimutagenicity in a Salmonella mutation assay. The present study focused on the effect of a low-polar extract of C. nervosum seed on the early stages of diethylnitrosamine (DEN)- and dimethylhydrazine (DMH)-induced carcinogenesis in rats. MethodsDried C. nervosum seed powder was extracted using dichloromethane. To study its effect on the initiation stage of carcinogenesis of rats, they were fed with various doses of C. nervosum seed extract (CSE) for 21 days. DEN injection was used to initiate hepatocarcinogenesis and partial hepatectomy was performed to amplify mutated hepatocytes resulting in micronucleated hepatocyte formation. To study the role of CSE on the promotion stage, rats were injected with DEN and DMH to induce preneoplastic lesions and the numbers of glutathione S-transferase placental form (GST-P) positive foci in the liver and aberrant crypt foci (ACF) in the colon were measured. This was followed by CSE administration for 10 weeks. The inhibitory mechanisms of CSE on initiation and promotion stages, including xenobiotic metabolism, cell proliferation and apoptosis, were investigated. ResultsThe total phenolic content in CSE was 80.34 ± 2.29 mg gallic acid equivalents (GAE) per g of extract and 2,4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone was found to be a major flavonoid. The main terpenoids in CSE were β-selinene, α-selinene, γ-selinene and o-cymene while 24(Z)-methyl-25-homocholesterol was a major phytosterol. CSE significantly decreased the number of micronucleated hepatocytes in DEN-initiated rats and enhanced the activities of hepatic glutathione S-transferase and UDP-glucuronyltransferase. Furthermore, the formation of preneoplastic lesions in the liver and colon was statistically reduced by CSE. CSE also diminished cell proliferation in the liver and colon indicated by the number of PCNA positive cells. However, CSE did not alter the numbers of apoptotic hepatocytes and colonocytes in DEN- and DMH-initiated rats. ConclusionsThe dichloromethane extract of C. nervosum seed demonstrated chemopreventive effects on chemically-induced carcinogenesis in both initiation and promotion stages in rats. The inhibitory mechanism might be involved in the modulation of hepatic detoxifying enzymes and suppression of hepatocyte and colonocyte proliferation.

MicroRNA-520c-3p targeting of RelA/p65 suppresses atherosclerotic plaque formation

Atherosclerosis is a chronic inflammatory disease, and it's the leading cause of death worldwide. Dysregulation of microRNAs (miRNAs) has been found to be associated with atherosclerosis. miR-520c-3p has been implicated in several types of cancer. However, little is known about the role of miR-520c-3p in atherosclerosis. In this study, we found that miR-520c-3p agomir decreased atherosclerotic plaque size, collagen content, the quantity of PCNA-positive cell and RelA/p65 expression of vascular smooth muscle cells (VSMCs) in the aortic valve of apoE-/- mice in vivo. The possible mechanisms of the protective effects of miR-520c-3p on atherosclerotic mice were then investigated in VSMCs. in vitro experiments showed that miR-520c-3p expressions were significantly reduced in human aortic vascular smooth muscle cell (HASMCs) treated with platelet-derived growth factor (PDGF-BB). miR-520c-3p mimics repress PDGF-BB-mediated the proliferation, migration and decrease in the percentage of cells in G2/M phase, which was associated with downregulation of RelA/p65. Mechanistically, miRNA pull-down, luciferase reporter and mRNA stability assays confirmed miR-520c-3p mimics was able to directly target 3'-UTR of RelA/p65 mRNA and decreased half-life of RelA/p65 mRNA in HASMCs. Overexpression of RelA/p65 reversed the inhibition of cell proliferation induced by miR-520c-3p mimics in HASMCs. In conclusion, our findings suggest that miR-520c-3p inhibits PDGF-BB-mediated the proliferation and migration of HASMCs by targeting RelA/p65, which may provide potential therapeutic strategies in atherosclerosis treatment.

Resveratrol Ameliorates the Seminiferous Tubules Damages Induced by Finasteride in Adult Male Rats.

Finasteride is commonly used in the management of alopecia and nodular prostatic hyperplasia. It was reported to have a harmful effect on spermatogenesis with subsequent infertility. Thus, this research was to determine the ameliorative effect of resveratrol against testicular damage caused by finasteride. Forty adult male rats were randomly divided into four main groups: group I acted as the control, group II was administrated resveratrol 20 mg/kg/day, group III was administrated finasteride 5 mg/kg/day, and group IV was administrated finasteride and resveratrol as in the previous groups. Finasteride induced a significant decrement in the testosterone and dihydrotestosterone levels. The level of malondialdehyde significantly increased, while the levels of glutathione peroxidase, superoxide dismutase, and catalase significantly decreased in the finasteride-administrated rats. Variable histopathological alterations in the testes were revealed in the form of irregular seminiferous tubules. Some seminiferous tubules appeared with degenerated germinal epithelium. Others showed detachment of their germinal epithelium. Congested blood vessels and homogeneous acidophilic substance in-between tubules were also detected. A significant decrement in PCNA positive cells and a significant increment in Bax expression were demonstrated. Ultrastructural examination showed Sertoli cells with rarefied cytoplasm. Vacuolated cytoplasm, shrunken nuclei, and dilated perinuclear spaces were also revealed in the spermatogonia, primary spermatocytes, and early spermatids. On the contrary, few changes were noticed in rats received resveratrol concomitant with finasteride. This study indicated that resveratrol exerted a potent ameliorative effect against testicular injury caused by finasteride.

Histological and Immunohistochemical Evaluation of the Ameliorating Role of Propolis and/or Intermittent Fasting on Induced Oral Mucositis in Albino Rats.

Context: Oral Mucositis (OM) is a very painful inflammatory condition, complicating patient’s life. Hence it was appealing to find natural remedies, dietary modifications or both to manage this condition successfully. Aim: to evaluate the efficiency of Propolis (P) and / or Intermittent Fasting (IF) in ameliorating OM. Materials & Methods: 40 male Albino rats weighing 200-250 grams, divided into: Negative Control Group (-ve control Gp): 8 rats kept healthy, Positive Control Group (+ve): 8 rats subjected to OM induction in their buccal mucosa, (P) Gp: 8 rats subjected to OM Induction, given Propolis orally, (IF) Gp: 8 rats subjected to OM induction & IF, (P&IF) Gp: 8 rats subjected to OM induction, maintained on both P &IF simultaneously. After three weeks, buccal mucosae were processed and stained by H&E, Masson’s Trichrome, and Anti-PCNA, examined by light microscope. Results: +ve control Gp showed significant increase in vacuolated epithelial cells, apparent decrease in immature collagen fibers in CT, and significant reduction in PCNA-positive cells. In the P, IF, and P&IF Gps, there were marked improvement in epithelium and CT. The (P&IF) Gp manifested maximum decrease in vacuolated epithelial cells, increase of immature collagen fibers, and maximum increase in PCNA positive cells. Discussion & conclusion: OM caused tissue destruction, which was evident in the +ve control Gp. Propolis intake greatly improved the histological appearance (confirming anti-inflammatory role of Propolis) as well as IF (confirming immune-boosting action). However, using both P & IF resulted in maximum improvement due to combined actions of both.

Increased myofiber size and reduced satellite cell numbers in medial rectus muscle of patients with intermittent exotropia

ABSTRACT Purpose To elucidate the differences in muscle bundle and satellite cells in medial rectus muscle through histological and Immunofluorescence studies of intermittent exotropia patients and normal controls. Materials and Methods: From January 2015 to December 2017, 15 patients who underwent medial rectus resection surgery at Kosin University Gospel Hospital were enrolled. Four medial recti muscles collected from two brain-dead men without strabismus were used as controls and compared with the intermittent exotropia group. Hematoxylin and eosin (HE) staining were performed, and all muscle bundle diameters were measured with the Image J program and compared to the mean value. Immunological staining for MyoHC (Myosin Heavy Chain), PAX7 (Transcription Factor), and PCNA (Proliferating Cell Nuclear Antigen) were performed to analyze the distribution of myocytes and PAX7-positive and PCNA-positive cells. Results: The mean ages of the strabismus and control groups were 17.33 ± 13.05 and 22.0 ± 5.85 years, respectively, and the male to female ratio was 7:2 and 2:0. The average angle of deviation in the exotropia patients was 36.0 ± 16.83 prism diopters. The mean muscle bundle diameter under light microscopy was 60.21 ± 1.48 in the exotropia group and 52.27 ± 0.74 in the control group. The exotropia group showed significantly greater mean muscle bundle diameter (p < .001) and diameter regularity than the control group. The PAX7(+)/muscle bundle number ratio was 0.016 ± 0.014 and 0.056 ± 0.015 in the exotropia group and control group, respectively (p < .001), and the PCNA(+)/muscular bundle number ratio was 0.015 ± 0.017 and 0.182 ± 0.102 (p < .001). Both were significantly higher in the control group compared to that in the exotropia patients. Conclusion: In the exotropia group, mean diameter of medial rectus muscle bundle was significantly larger than that of control group. The ratios of PAX7 and PCNA to muscle bundle number were significantly higher in the control group than intermittent extropia group. We found the negative relationship between activation of satellite cells and muscle bundle diameter, and it might take one step forward to elucidate the pathogenesis of intermittent extropia.

Antiapoptotic and antioxidant capacity of phytochemicals from Roselle (Hibiscus sabdariffa) and their potential effects on monosodium glutamate-induced testicular damage in rat.

Monosodium glutamate (MSG), common flavor enhancer and feed additive, causes male reproductive dysfunction. However, Roselle tea, popular Hibiscus sabdariffa (HS) beverage, has a controversial effectiveness on male fertility. Therefore, the current study aimed to investigate either the adverse effect of aqueous HS extract (HSE) on the testicle or its potential ameliorative role including some stress markers, biochemical and immunohistochemical expressions in rats subjected to MSG. Here, the animals were divided into four groups that were given distilled water, HSE, MSG, and HSE + MSG respectively via gavage. After 6weeks from the beginning of experiment, blood samples were collected for hormonal analysis. Additionally, testicular specimens were excised and processed for oxidative/antioxidant parameters determination, histological examination, and immunohistochemical evaluation of Bax and PCNA positive spermatogenic cells. Preliminary phytochemical analyses as well as antioxidant capacity of the HSE were tested. Our results revealed a strong inhibitory activity of the HSE phytochemical constituents against DPPH radical. MSG group revealed a significant decrease of testosterone, LH, FSH, and antioxidant parameters with elevated MDA compared with control and HSE groups. Additionally, an alteration of the testicular histo-architecture was observed among MSG group along with increased Bax and decreased PCNA positive cells. Meanwhile, the HSE showed a potent protective effect against testicular damage as well as oxidative stress induced by MSG. On the whole, our findings provide evidence that HSE can ameliorate MSG-induced testicular toxicity via induction of cell proliferation along with reduction of oxidative stress and cellular apoptosis in adult rat that attributed to the antioxidant and antiapoptotic effects of its phytochemical constituents.

Expression of Notch-Hif-1α signaling pathway in liver regeneration of rats.

ObjectiveTo investigate whether the Notch–Hif-1α signaling pathway is involved in liver regeneration.MethodsRats were divided into two groups and treated with daily intraperitoneal injections of saline (control) or the gamma-secretase inhibitor, Fli-06, for 2 days. Two-thirds of the rat livers were resected and rats were later euthanized at specific time points post-resection to analyze the remnant livers. Each group's liver/body weight ratio was calculated, and immunostaining and western blotting were used to determine the cell proliferation marker, PCNA and Ki-67 expression. Real-time PCR and western blotting were used to compare the mRNA expression of Notch homolog-1 (Notch1), hairy and enhancer of split-1 (Hes1), and vascular endothelial growth factor (Vegf), and the protein expression of NICD and HIF-1α, respectively.ResultsThe liver/body weight ratios and number of Ki-67- and PCNA-positive cells were significantly lower in the experimental group than the control group, indicating lower levels of liver regeneration following the disruption of Notch signaling by Fli-06. The Hes1 and Vegf mRNA levels and NICD and HIF-1α protein expression levels were all down-regulated by Fli-06 treatment.ConclusionNotch–Hif-α signaling pathway activation plays an important role in liver regeneration, where it may contribute toward liver cell proliferation.

Uncovering the mechanism of Jueyin granules in the treatment of psoriasis using network pharmacology

Ethnopharmacological relevanceOur clinical practice demonstrated that Jueyin granules (JYG) benefit patients with mild to moderate psoriasis vulgaris without apparent adverse effects. JYG have been shown to inhibit epidermal proliferation in an imiquimod (IMQ)-induced psoriasis-like mouse model, as well as keratinocyte proliferation. Moreover, JYG causes no acute or chronic toxicity in animal models. However, its related molecular mechanism has still not been elucidated. Aim of the studyTo assess the mechanism of JYG against psoriasis. Materials and methodsThis study combined network pharmacology analysis with experiments to investigate the mechanism of JYG against psoriasis. First, the molecular docking technology was used to construct the network of medicinal materials-core active plant ingredients-core targets and identify possible drug targets. Next, high-performance liquid chromatography (HPLC) was used for quality control of JYG. Finally, a mice model of psoriasis was used to further verify the effects of JYG. Results(1) Molecular docking analysis of network pharmacology revealed that the therapeutic effects of JYG on psoriasis might be achieved through Vitamin D Receptor (VDR) effects. (2) The concentrations of chlorogenic acid and paeoniflorin were determined using HPLC to establish quality control of JYG. (3) JYG ameliorated pathological characteristics that included in vivo reductions in erythema, scale, and infiltration scores of back and ear lesions in IMQ-induced psoriasis-like mice. Moreover, a reduced number of PCNA-positive and Ki67-positive cells were observed in the epidermis of JYG-treated lesions. JYG also reduced inflammation (interleukin (IL)-17, IL-23) in the peripheral blood of IMQ-induced psoriasis-like mice. As expected, JYG was found to upregulate VDR expression and downregulate p-STAT3 expression in the IMQ group, which may contribute to its mechanism against psoriasis. ConclusionOverall, this study clarifies the mechanism of JYG against psoriasis and provides evidence to support its clinical use.

Surgical results and pathological analysis of cardiac fibroma in the adolescent and the adult.

Disparities may exist between the adolescent and the adult patients with cardiac fibromas in the symptoms, surgical outcomes, and pathological characteristics. The aim of this study was to compare short and midterm surgical outcomes of cardiac fibromas and to compare the biomarker expressions of tumor tissue samples between the adult and the adolescent. Consecutive patients with the diagnosis of cardiac fibroma were admitted and received surgeries. Primary outcomes included in-hospital mortality, low cardiac output, and readmission due to heart failure. The expression of PCNA and Ki67, two widely adopted indicators of cell proliferation, were evaluated in tissue samples. A total of five adolescent patients and five adult patients diagnosed as cardiac fibroma were admitted and given surgeries. When compare with the adults, the adolescent patients were more likely to present symptoms on admission (P = .048). Postoperative low cardiac output syndrome was significantly higher in the adolescents than in the adults (80.0% vs 0.0, P = .048). The tumor volume relative to ventricular end diastolic diameter had good discriminative ability for low cardiac output (c statistics: 0.96). Pathologically, the percentage of PCNA-positive cell nuclei was significantly higher in the adolescents than in the adults (36.04% ± 10.54% vs 4.15% ± 3.93%, P = .001). However, there were no Ki67-positive nuclei in the 10 cases. In the current study, we found that postoperative low cardiac output was more likely to occur in the adolescent patients than in the adult patients. When compared with the adult patients, significantly more PCNA-positive nuclei were observed in the adolescents.

Modulatory Role of Melatonin on the Structure of Hard Palatal Mucosa of Aged Rats

Introduction: Aging greatly diminishes the ability of oral tissues to withstand various types of functional insults, and reduces cell regeneration. Hence, anti-oxidation is mandatory to preserve oral tissues especially while aging. Melatonin is an anti-oxidant with established potential in hindering age-related changes. Aim: to evaluate the extent of anti-oxidant and anti-inflammatory potentials of melatonin on aged hard palatal tissues of rats. Materials & Methods: twenty-one Albino rats 200-250 gms each, were used in this study. They were equally divided into 3 groups; Control Young Gp: 6 months of age, Control Senile Gp:18 months of age, and Melatonin Gp: 18 months of age. After 8 weeks, rats were sacrificed by high dose of anesthesia. The hard-palatal mucosae of all rats were separated and processed for histologic examination by H&E, Masson Trichrome stain, and immuno-histochemical staining by Anti-PCNA, then examined by light microscopy. Results: the control senile Gp showed statistically significant increase in vacuolated epithelial cells and area percentage of mature collagen fibers, as well as statistically significant reduction in PCNA-positive cells and apparent reduction of epithelial thickness. On the other hand, the melatonin Gp displayed statistically significant increase in PCNA-positive cells, area percentage of immature collagen fibers, while significant decrease in No. of vacuolated epithelial cells and apparent increase of epithelial thickness. Discussion: Aging resulted in deteriorated regeneration, enhanced apoptosis, with reduced cellularity and vascularity. On the other hand, melatonin greatly improved cellular proliferation, reduced apoptosis, enhanced CT tissue regeneration and collagen production. As well as reduction in inflammatory cells.

Role of TNF-α and FGF-2 in the Fracture Healing Disorder of Type 2 Diabetes Model Induced by High Fat Diet Followed by Streptozotocin.

PurposeTo investigate the effect of TNF-α and FGF-2 in the fracture healing disorder of type 2 diabetes.Design/Methodology/ApproachRat diabetes-bone traction model was established to investigate the effect of type 2 diabetes on the fracture healing and the association of TNF-α and FGF-2 with the process. Serological examination was performed to detect the related diabetes indexes. The proliferation activity of the cells was detected by MTT assay. The expressions of FGF-2 and TNF-α of osteoblasts in high glucose culture environment were detected by histochemistry and Western blotting.FindingsSerological examination showed that in rats fed with high fat and sugar diet for 8 weeks, the serum total cholesterol (TC), triglyceride (TG), fasting insulin (FINs) significantly increased, but fasting blood glucose (FBG) had no significant change. Two weeks after intraperitoneal injection of STZ, rat serum TG, TC, and FBG increased significantly, while FINs did not change obviously. Two weeks after traction osteogenesis, X-ray examination and HE staining showed that the area of osteotylus in the diabetes group was significantly smaller than that in the control group. The number of PCNA positive cells in the osteotylus of diabetes group was significantly decreased. In the osteotylus of diabetes group, the expression of TNF-α was significantly increased and the expression of FGF-2 was significantly decreased. MTT assay showed that the proliferation activity of MC3T3-E1 cells in high glucose culture medium groups was significantly decreased at 24th hour of the culture, compared with the normal culture medium group. qPCR results showed that the expression of FGF-2 was significantly decreased while the expression of TNF-α was significantly increased in high glucose culture medium groups.Originality/ValueIt was concluded that type 2 diabetes mellitus affected the fracture healing by causing osteoblast proliferation disorder. TNF-α and FGF-2 were important related factors for the process.

Effect of photodynamic therapy on the morphological changes of periapical inflammation: An experimental study in rats.

We aimed to explore the morphological changes with photodynamic therapy (PDT) in experimentally produced periapical lesions in rats, and to investigate if PDT used as an adjunct to conventional root canal debridement helps to enhance healing the inflammatory response around root apex of rats. Sixty adult Sprague-Dawley male rats (70-90 days/140-160 g) were experimented. Pulp was removed from the mesial root of the first maxillary molar and left open to oral environment to allow the formation of periapical lesion. The rats were numbered and randomly divided into two groups: (i) PDT group (n = 30) received a single session of PDT with conventional debridement and, (ii) Control group (n = 30) received conventional debridement but with no PDT. PDT application involved the use of methylene blue photosensitizer for 5 min inside the root canals and irradiated with diode laser of 805 nm and 20 W power output for 90 s and sealed. After 4 weeks, the experimental rats were sacrificed by cervical dislocation. The maxillary first molar was then collected along with the surrounding tissue for further processing. Hematoxylin and eosin and immunohistochemical staining were used to observe the morphological effects. Proliferating Cell Nuclear Antigen (PCNA), STRO-1 and CD-44 were used as the primary antibodies for the immunohistochemical study. A reduction in inflammatory cells, which were mainly composed of lymphocytes, was observed in the periapical lesions after PDT. The number of PCNA-positive cells increased to approximately twice in the PDT as compared to the control group. These PCNA-positive cells included STRO-1 and CD-44 positive cells, indicating enhancement of wound healing and reduction in inflammatory cells. The findings of the present experimental study indicate that PDT application induced proliferation of PCNA-positive cells, which included STRO-1 and CD44-positive cells. This suggests that PDT may help to enhance healing periapical lesion, indicating the potential of PDT in the treatment of periapical periodontitis.

Expression of CD34, PCNA and VEGF in CIN-3 and Invasive Squamous Cell Carcinoma Cervix: A Pilot Study

Although the role of classical (clinico-pathological features) and modern prognostic factors (immune biomarkers) has been described in pathogenesis of squamous cell carcinoma cervix (SCCC), the role of VEGF and PCNA-positive endothelial cells in CD34 expressing blood vessels has been a subject of great interest and has not been validated yet. So, the present pilot study was aimed to study the expression of VEGF and PCNA-positive cells in CD34 positive blood vessels in “cervical intraepithelial lesion-3” (CIN-3) and “invasive” areas of SCCC. It was a cross-sectional observational pilot study. In this study, 100 hysterectomy specimens done for SCCC were screened. Immunohistochemical expression of VEGF and PCNA-positive endothelial cells in CD34 expressing blood vessels was studied on 21 cases which showed both CIN-3 and invasive areas. In “invasive” areas, epithelial component had higher expression of VEGF with 61.9% and 38.1% cases showing grades 2 and 3 staining, respectively. 100% stromal component showed grade 1 in “CIN-3” areas, and 90.4% showed grade 2 in invasive cases. In case of VEGF expression in “CIN-3” compartment, epithelial compartment had grade I in 57.1% cases and grade II expression in 42.9% cases. PCNA expression was up to grade 1 in most “CIN-3” areas, whereas higher grades (2 or 3) were seen in “invasive” compartments. VEGF expression is more in “invasive” front of the tumour. The tumour parenchyma rather than the tumour stroma is responsible for neovascularisation. The vessels at the periphery on “invasive” front of tumour proliferate more.

The inhibitory influence of toluene on mare ovarian granulosa cells can be prevented by fennel

The influence of environmental contaminant toluene and of plant fennel (Foeniculum vulgare Mill.) on reproduction are reported, but the mechanisms of their action and the protective effect of fennel on contaminant influence remain to be elucidated. In this study, we hypothesized that toluene and fennel directly affects basic ovarian cell functions, and that fennel can be used as an appropriate natural protective agent against the potential adverse effects of toluene. This study aimed to examine the action of toluene (20 μg/mL) and fennel extract (0, 1, 10, 100 μg/mL), and assess their combination on viability, proliferation, apoptosis, and hormone release by cultured healthy mare ovarian granulosa cells. Viability, proliferation (percentage of PCNA-positive cells), apoptosis and release of progesterone, oxytocin and prostaglandin F were evaluated by using Trypan blue exclusion tests, immunocytochemistry and enzyme immunoassays, respectively. Toluene, when given alone, inhibited viability, proliferation, apoptosis, progesterone, prostaglandin F and IGF-I. However, it did not affect oxytocin release. Moreover, Fennel, when given alone, inhibited viability, progesterone, and prostaglandin F release, as well as stimulating proliferation and oxytocin release. In addition, Fennel did not affect apoptosis. When given in combination with toluene, fennel was able to suppress, and even invert, the effects of toluene on viability, proliferation, apoptosis, prostaglandin F, and IGF-I. However, it did not alter its effect on progesterone release. Moreover, fennel induced the inhibitory effect of toluene on oxytocin output. The findings of our study suggest direct adverse effects of toluene on the basic ovarian functions of mares. Lastly, we also observed the direct influence of fennel on these functions, as well as its ability to be a natural protector against the action of toluene on the ovarian functions of mares.

Factors secreted from dental pulp stem cells show multifaceted benefits for treating experimental temporomandibular joint osteoarthritis

Temporomandibular joint osteoarthritis (TMJOA) is a degenerative disease characterized by progressive cartilage degeneration, abnormal bone remodeling, and chronic pain. In this study, we aimed to investigate effective therapies to reverse or suppress TMJOA progression. To this end, we performed intravenous administration of serum free conditioned media from human exfoliated deciduous teeth stem cells (SHED-CM) into a mechanical-stress induced murine TMJOA model. SHED-CM administration markedly suppressed temporal muscle inflammation, and improved bone integrity and surface smoothness of the destroyed condylar cartilage. Moreover, SHED-CM treatment decreased the number of IL-1β, iNOS, and MMP-13 expressing chondrocytes, whereas it specifically increased PCNA-positive cells in the multipotent polymorphic cell layer. Notably, the numbers of TdT-mediated dUTP nick end labeling (TUNEL)-positive apoptotic chondrocytes in the SHED-CM treated condyles were significantly lower than in those treated with DMEM, whereas the proteoglycan positive area was restored to a level similar to that of the sham treated group, demonstrating that SHED-CM treatment regenerated the mechanical-stress injured condylar cartilage and subchondral bone. Secretome analysis revealed that SHED-CM contained multiple therapeutic factors that act in osteochondral regeneration. Our data demonstrated that SHED-CM treatment promoted the regeneration and repair of mechanical-stress induced mouse TMJOA. Our observations suggest that SHED-CM has potential to be a potent tissue-regenerating therapeutic agent for patients with severe TMJOA.

ARMC5 mutations are associated with high levels of proliferating cell nuclear antigen and the presence of the serotonin receptor 5HT4R in PMAH nodules.

Objective To analyze the morphological and functional characteristics of primary macronodular adrenal hyperplasia (PMAH) nodules carrying or not carrying ARMC5 mutations and the consequences of the presence of mutations in terms of the pattern of macronodule composition and functional state. Subjects and methods The analyses were performed by hematoxylin-eosin staining, immunohistochemistry, microdissection of spongiocyte tissue and RT-qPCR of histological sections from 16 patients diagnosed with PMAH with germline (5) or germline/somatic mutations (5) and without mutations (6) in the ARMC5 gene. Results Hyperplastic nodules were predominantly composed of spongiocytes in mutated and nonmutated sections. ARMC5 mRNA expression in spongiocytes was higher in ARMC5-mutated nodules than in ARMC5-nonmutated nodules, and homogenous ARMC5 protein distribution was observed. The presence of arginine-vasopressin receptor (AVP1AR) and ectopic ACTH production were observed in both cell populations regardless of ARMC5 mutations; the numbers of serotonin receptor (5HT4R)- and proliferating cell nuclear antigen (PCNA)-positive cells were higher in macronodules carrying ARMC5 mutations than in those without mutations. Conclusions Our results suggest that the presence of ARMC5 mutations does not interfere with the pattern of distribution of spongiocytes and compact cells or with the presence of AVP1AR, gastric-inhibitory polypeptide receptor (GIPR) and ectopic ACTH. Nevertheless, the higher numbers of PCNA-positive cells in mutated nodules than in nonmutated nodules suggest that mutated ARMC5 can be related to higher proliferation rates in these cells. In conclusion, our results provide more information about the crosstalk among abnormal GPCRs, ectopic ACTH in steroidogenesis and the ARMC5 gene, which may be relevant in understanding the pathogenesis and diagnosis of patients with PMAH.

Resveratrol prevented experimental pulmonary vascular remodeling via miR-638 regulating NR4A3/cyclin D1 pathway

ObjectiveResveratrol has shown benefit for pulmonary hypertension improvement. Our previous reports showed NR4A3/cyclin D1 pathway promoted pulmonary arterial smooth muscle cells (PASMCs) proliferation. This study tried to explore the mechanism underlying this process, focusing on the role of resveratrol in regulation of miRNA and NR4A3. MethodsRats were injected with monocrotaline (MCT) to establish pulmonary hypertension (PH) models. Resveratrol was used to prevent pulmonary vascular remodeling. Primary rat PASMCs were cultured in vitro and stimulated by platelet-derived growth factor (PDGF) with or without resveratrol. Cells proliferation and expression of miR-638 as well as NR4A3 were evaluated. ResultsMCT resulted in significant pulmonary vascular remodeling and down-regulation of miR-638, which could be suppressed by resveratrol. Moreover, PDGF-induced PASMC proliferation and miR-638 down-regulation were both significantly prevented by resveratrol treatment in vitro. MiR-638 mimics markedly inhibited PASMC proliferation and percentage of PCNA-positive cells in vitro. But anti-miR-638 could markedly promote cells proliferation and percentage of PCNA-positive cells. The luciferase reporter assay showed that NR4A3 was a direct target of miR-638. The loss-of-function and gain-of-function experiments indicated that NR4A3 promoted proliferation via cyclin D1 pathway. ConclusionOur data indicated that resveratrol prevented MCT-induced pulmonary vascular remodeling via miR-638 regulating NR4A3/cyclin D1 pathway.