a-simplified-efficient-method-to-isolate-primary-human-keratinocytes

A Simplified and Efficient Method to Isolate Primary Human Keratinocytes from Adult Skin Tissue

Primary human keratinocytes isolated from fresh skin tissues and their expansion in vitro have been widely used for laboratory research and for clinical ...

localizing-function-specific-targets-for-transcranial-magnetic

Localizing Function-specific Targets for Transcranial Magnetic Stimulation in the Absence of Navigation Equipment

Repetitive transcranial magnetic stimulation (rTMS) is a non-invasive technique that modulates neural activity in the brain. Studies have shown that rTMS ...

Shandong University

Chengdu Sport University

To establish a model of acute lung injury induced by paraquat poisoning and to observe the effects of anticoagulant therapy on acute lung injury induced by paraquat poisoning.

[Experimental anticoagulant therapy of acute lung injury induced by paraquat].

In this report, we present a twenty-nine year old patient with a five-year history of Behcet's disease (BD), who developed an ascending aortic aneurysm. Chest enhanced computed tomography scanning demonstrated a saccular aneurysm of the ascending aorta, with a maximum diameter of 11 cm. The patient was administered with prednisone, thalidomide and anticoagulant. A regular follow up was carried out. The patient lived well with blood sedimentation and C-reactive protein in the normal range.

Recurrent aortic aneurysm due to Behcet's disease: a case report from China.

To investigate the clinical significance of dynamic changes of blood urea nitrogen (BUN), serum creatinine (Cr), serum cystatin (Cys C) and urinary protein on renal injury with paraquat poisoning.

[Clinical studies of dynamic changes on the renal injury indicators of acute paraquat poisoning].

Suicidal poisoning by ingestion of organophosphate (OP) insecticides represents a serious emergency with a high mortality rate. However, attempted suicide via the parenteral route has rarely been reported. The present study reports a case of parasuicide by self-injection of an organophosphorus insecticide (phoxim, phenylglyoxylonitrile oxime O,O-diethyl phosphorothioate) into the distal region of the left arm. The patient developed necrosis at the injection site and an abscess of the affected limb following injection. A fasciotomy and surgical debridement resulted in the symptoms of the patient disappearing within a few days and were vital in shortening the course of the disease.

Parasuicidal poisoning by intramuscular injection of insecticide: A case report.

Paraquat poisoning is well known for causing multiple organ function failure (MODS) and high mortality. Acute lung injury and advanced pulmonary fibrosis are the most serious complications. Bosentan is a dual endothelin receptor antagonist. It plays an important role in treating PF. There is no related literature on the use of bosentan therapy for paraquat poisoning.

Using bosentan to treat paraquat poisoning-induced acute lung injury in rats.

The present report describes two cases of paraquat poisoning by skin absorption. The cases involved contractual workers who were spraying paraquat in an orchard. Whilst spraying, some solution adhered to their skin. The skin developed erythema followed by blistering and hemorrhaging hemorrhagic diabrosis. Six days later the patients were admitted to the Department of Poisoning and Occupational Disease, Qilu Hospital of Shandong University (Jinan, China) with 3 and 2% total body surface area (TBSA) burns, respectively. Surgical debridement was performed and immunosuppressants were administered during the patients' treatment. The patients were treated successfully and had made a complete recovery following 21 days. From these cases it was examined how paraquat may cause skin injuries and occasionally poisoning. To the best of our knowledge, cases of paraquat poisoning are rare in China. A review of the relevant literature was performed.

Paraquat poisoning by skin absorption: Two case reports and a literature review.

In China, and other Asian countries, numerous patients have succumbed to pulmonary fibrosis induced by paquarat poisoning, but the early pathogenesis remains unclear. In this study the effect of cytokine transforming growth factor (TGF)-β1 was observed in early acute paraquat poisoning and examined the mechanism by which paraquat caused early acute lung injury. It was discovered that the rat serum TGF-β1 levels in the paraquat groups were significant higher than that in the control group (P<0.05) and the rat pulmonary TGF-β1 mRNA expression levels were also higher than that in the control group (P<0.05). Histological examination indicated that the rat lung tissue was broad and congested, and had been infiltrated by inflammatory cells. Masson's trichrome staining for collagen showed that the lung tissue appeared fibrotic following paraquat poisoning. Ultramicrostructure observation found that macrophages, red blood cells, lymphocytes and granulocytes infiltrated the alveolar space and there were cytolysosomes in the macrophages. The shape of the type II alveolar epithelial cell nuclei were irregular with karyopyknosis. The heterochromatin migrated to the cell edge and lamellar body vacuolization was also observed. Type I alveolar epithelial cells shrank. In conclusion, the effect of cytokine TGF-β1 on paraquat-induced acute lung tissue injury may be important.

Effect of transforming growth factor-β1 on acute lung injury caused by paraquat.

Overexpression of integrin β6 plays an important role in a variety of malignant tumor invasion and metastasis.

Clinical significance of integrin β6 as a tumor recurrence factor in follicular thyroid carcinoma.

We tested whether the a simple injection known as the patch assay could reconstitute mature hair follicles by culture-expanded human cells and explored whether the assay could reflect the trichogenicity of cultured cells.

The patch assay reconstitutes mature hair follicles by culture-expanded human cells.

Primary skin epidermal cells isolation and in vitro expansion culture have been widely used for laboratory research and clinical applications. The conventional methods involving sequential enzymatic digestion of adult tissues have given low cell recovery rate and reduced cell viability. We report here an advanced method for human primary epidermal progenitor cells isolation from skin tissues including the Rho kinase inhibitor Y-27632. Compared with traditional protocols, the current protocol is simple, easy, and faster; moreover, it gives a greater yield of integrin-expressing epithelial stem cells. In addition, our new methodology does not require a separation of epidermis from dermis because the medium selectively blocks focal adhesion and growth of dermal cells. Importantly, the cells isolated from this method can maintain their regeneration potential and quickly reconstitute a mature human skin in vivo after grafting onto nude mice. In brief, we describe here a simple (one step) and serum-free method for isolating primary epidermal stem cells from adult tissues. The isolated cells may be widely used for both laboratory studies and clinical application, especially in the field of tissue engineering and regeneration.

Y-27632 simplifies the isolation procedure of human primary epidermal cells by selectively blocking focal adhesion of dermal cells.

Remodeling of maternal spiral artery after the implantation of embryo relies on tightly regulated trophoblast functions and is pivotal to ensure the success of pregnancy. Low-density lipoprotein receptor-related protein 6 (LRP6) is implicated in angiogenesis and several vascular diseases, but its function in trophoblast regulation is still not fully understood. We aimed to investigate the involvement of LRP6 in trophoblast functions during maternal spiral artery remodeling. HTR-8/SVneo cells, a trophoblast cell line, were employed to examine the impact of LRP6 knockdown on proliferation, apoptosis, and migration, as well as invasion and tube formation. Expression of matrix metalloproteinases (MMPs), angiogenic factors placental growth factor (PlGF), and vascular endothelial growth factor (VEGF), as well as activation of canonical Wnt/β-catenin signaling pathway, which was involved in artery remodeling process, was also analyzed. Lipoprotein receptor-related protein 6 knockdown suppressed proliferation, migration, invasion, and tube formation, as well as induced apoptosis in HTR-8/SVneo cells. In addition, LRP6 knockdown also significantly decreased expression of MMPs, PlGF, and VEGF and activation of Wnt/β-catenin signaling pathway. Lipoprotein receptor-related protein 6 might positively regulate the remodeling of the maternal artery through both endovascular formation and trophoblast invasion.

Low-Density Lipoprotein Receptor-Related Protein 6 Is Essential for Trophoblast Survival and Invasion.

Stem cells play a crucial role in maintaining and repairing tissues during homeostasis and following injury. The efficient procurement of high quantity and quality of skin stem cells is important for both laboratory studies and clinical applications. Here, we describe a one-step isolation procedure to efficiently obtain both epidermal and dermal cell population from human skin specimen, based on the different influence of the Rho kinase inhibitor Y27632 on the growth of epidermal and dermal cells during the initial culture. Compared with the conventional methods, our protocol shows that it is simpler and less time consuming and can efficiently obtain the high quality of skin stem cells and can maintain the stem cell features after culture expansion.

One-Step Simple Isolation Method to Obtain Both Epidermal and Dermal Stem Cells from Human Skin Specimen.

An increased incidence of skin inflammatory diseases is frequently observed in organtransplanted patients being treated with calcineurin inhibitor-based immunosuppressive agents. The mechanism of increased skin inflammation in this context has however not yet been clarified. Here we report an increased inflammation following inhibition of calcineurin signaling seen in both chemically induced mouse skin tumors and in tumors grafted from H-rasV12 expressing primary human keratinocytes (HKCs). Following UVB or TPA treatment, we specifically found that deletion of the calcineurin gene in mouse keratinocytes (MKCs) resulted in increased inflammation, and this was accompanied by the enhanced production of pro-inflammatory cytokines, such as TNFα, IL-8 and CXCL1. Furthermore, expression of the RNA-binding protein, tristetraprolin (TTP) was down-regulated in response to calcineurin inhibition, wherein TTP was shown to negatively regulate the production of pro-inflammatory cytokines in keratinocytes. The induction of TTP following TPA or UVB treatment was attenuated by calcineurin inhibition in keratinocytes, and correspondingly, disruption of calcineurin signaling down-regulated the amounts of TTP in both clinical and H-rasV12-transformed keratinocyte tumor models. Our results further demonstrated that calcineurin positively controls the stabilization of TTP in keratinocytes through a proteasome-dependent mechanism. Reducing the expression of TTP functionally promoted tumor growth of H-rasV12 expressing HKCs, while stabilizing TTP expression counteracted the tumor-promoting effects of calcineurin inhibition. Collectively these results suggest that calcineurin signaling, acting through TTP protein level stabilization, suppresses keratinocyte tumors by downregulating skin inflammation.

The ARE-binding protein Tristetraprolin (TTP) is a novel target and mediator of calcineurin tumor suppressing function in the skin.

Rho Kinase (ROCK) plays crucial roles in the proliferation and migration of different type cells. ROCK inhibitor Y-27632 was previously reported to inhibit melanoma cell growth, and ROCK signaling was suggested to be therapeutic target for treating melanoma. However, the negative effect of Y-27632 on melanoma cells was mainly seen from studies on murine B16 melanoma cells. Here we reported that ROCK inhibitor actually promoted human melanoma cell growth and migration in vitro. Y-27632 increased the growth and migration of BRAF mutated melanoma cells, but had negative effect on wildtype melanoma cells or primary melanocytes. We discovered Y-27632 enhanced the growth of BRAF mutated melanoma cells through increased ATK and ERK activity. The in vivo study further confirmed that in vitro finding. These data suggested the effect of ROCK inhibitor on melanoma cells is cell-context dependent, and the application of ROCK inhibitor in treatment of melanoma needs further studied. This article is protected by copyright. All rights reserved.

ArticleTotal : 2	Year
Publication title	2018
Publication title	2025

Article	Year
[Experimental anticoagulant therapy of acute lung injury induced by paraquat]. Zhonghua lao dong wei sheng zhi ye bing za zhi = Zhonghua laodong weisheng zhiyebing zazhi = Chinese journal of industrial hygiene and occupational diseases\| PubMed ID: 22804886	2012
Recurrent aortic aneurysm due to Behcet's disease: a case report from China. Annals of thoracic and cardiovascular surgery : official journal of the Association of Thoracic and Cardiovascular Surgeons of Asia\| PubMed ID: 22971707	2013
[Clinical studies of dynamic changes on the renal injury indicators of acute paraquat poisoning]. Zhonghua lao dong wei sheng zhi ye bing za zhi = Zhonghua laodong weisheng zhiyebing zazhi = Chinese journal of industrial hygiene and occupational diseases\| PubMed ID: 23257035	2012
Parasuicidal poisoning by intramuscular injection of insecticide: A case report. Experimental and therapeutic medicine\| PubMed ID: 24137249	2013
Using bosentan to treat paraquat poisoning-induced acute lung injury in rats. PloS one\| PubMed ID: 24155875	2013
Paraquat poisoning by skin absorption: Two case reports and a literature review. Experimental and therapeutic medicine\| PubMed ID: 24250726	2013
Effect of transforming growth factor-β1 on acute lung injury caused by paraquat. Molecular medicine reports\| PubMed ID: 24535699	2014
Clinical significance of integrin β6 as a tumor recurrence factor in follicular thyroid carcinoma. Head & neck\| PubMed ID: 24844802	2015
The patch assay reconstitutes mature hair follicles by culture-expanded human cells. Regenerative medicine\| PubMed ID: 28749726	2017
Y-27632 simplifies the isolation procedure of human primary epidermal cells by selectively blocking focal adhesion of dermal cells. Journal of tissue engineering and regenerative medicine\| PubMed ID: 28752652	2018
Low-Density Lipoprotein Receptor-Related Protein 6 Is Essential for Trophoblast Survival and Invasion. Reproductive sciences (Thousand Oaks, Calif.)\| PubMed ID: 29439617	2018
One-Step Simple Isolation Method to Obtain Both Epidermal and Dermal Stem Cells from Human Skin Specimen. Methods in molecular biology (Clifton, N.J.)\| PubMed ID: 29478135	2018
The ARE-binding protein Tristetraprolin (TTP) is a novel target and mediator of calcineurin tumor suppressing function in the skin. PLoS genetics\| PubMed ID: 29723192	2018
ROCK inhibitor enhances the growth and migration of BRAF mutant skin melanoma cells. Cancer science\| PubMed ID: 30168234	2018

Qian Zhou

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