BACKGROUND: Increasing evidence indicates that oxidative stress and interferon γ (IFNγ)-driven cellular immune responses are responsible for the pathogenesis of vitiligo. However, the connection between oxidative stress and the local production of IFNγ in early vitiligo remains unexplored. The aim of this study was to identify the mechanism underlying the production of IFNγ by mast cells and its impact on vitiligo pathogenesis. METHODS: Skin specimens from the central, marginal, and perilesional skin areas of active vitiligo lesions were collected to characterize changes of mast cells, CD8 + T cells, and IFNγ-producing cells. Cell supernatants from hydrogen peroxide (H 2 O 2 )-treated keratinocytes (KCs) were harvested to measure levels of soluble stem cell factor (sSCF) and matrix metalloproteinase (MMP)-9. A murine vitiligo model was established using Mas-related G protein-coupled receptor-B2 (MrgB2, mouse ortholog of human MrgX2) conditional knockout (MrgB2 -/- ) mice to investigate IFNγ production and inflammatory cell infiltrations in tail skin following the challenge with tyrosinase-related protein (Tyrp)-2 180 peptide. Potential interactions between the Tyrp-2 180 peptide and MrgX2 were predicted using molecular docking. The siRNAs targeting MrgX2 and the calcineurin inhibitor FK506 were also used to examine the signaling pathways involved in mast cell activation. RESULTS: IFNγ-producing mast cells were closely aligned with the recruitment of CD8 + T cells in the early phase of vitiligo skin. sSCF released by KCs through stress-enhanced MMP9-dependent proteolytic cleavage recruited mast cells into sites of inflamed skin (Perilesion vs . lesion, 13.00 ± 4.00/high-power fields [HPF] vs . 26.60 ± 5.72/HPF, P <0.05). Moreover, IFNγ-producing mast cells were also observed in mouse tail skin following challenge with Tyrp-2 180 (0 h vs . 48 h post-recall, 0/HPF vs . 3.80 ± 1.92/HPF, P <0.05). The IFNγ + mast cell and CD8 + T cell counts were lower in the skin of MrgB2 -/- mice than in those of wild-type mice (WT vs . KO 48 h post-recall, 4.20 ± 0.84/HPF vs . 0.80 ± 0.84/HPF, P <0.05). CONCLUSION Mast cells activated by MrgX2 serve as a local IFNγ producer that bridges between innate and adaptive immune responses against MCs in early vitiligo. Targeting MrgX2-mediated mast cell activation may represent a new strategy for treating vitiligo.
Vitiligo/metabolism* ; Mast Cells/immunology* ; Animals ; Interferon-gamma/metabolism* ; Mice ; Humans ; Melanocytes/metabolism* ; Receptors, G-Protein-Coupled/genetics* ; Mice, Knockout ; Mice, Inbred C57BL ; Male ; Female ; Matrix Metalloproteinase 9/metabolism* ; Stem Cell Factor/metabolism*

The electromagnetic spectrum emitted by the sun includes visible light, infrared, and ultraviolet (UV). Among them, UV is one of the hotspots of research. Previous studies have paid more attention to the impact of UV on the skin because UV is one of the causes of skin damage. The physiological effects of UV on the skin are well understood, but its impacts on other organs of the body and other human diseases are unclear yet. This review pointed to the benefits of UV exposure in reducing weight gain, metabolic dysfunction, and cardiovascular disease. In addition, UV exposure may be helpful in reducing the incidence of diseases such as diabetes and colitis. UV may also play a role in inhibiting the development of myopia and depression. These findings provide new ideas for applying UV in the treatment of human diseases in the future. This paper reviewed the impacts of UV exposure on human non-skin diseases and explored the possibility of damage of acute UV exposure to organs, not only at the skin level, clarified the benefits and harms of UV for human body, and provided theoretical reference and research directions for deep UV exploitation and UV dose control.

To construct a eukaryotic expression plasmid containing the luciferase reporter gene (Fluc) to quickly detect apoptosis. Four amino acids, Asp-Glu-Val-Asp (DEVD), the recognize motif of Caspase-3, were introduced into the middle of the Fluc-C and N fragment. Meanwhile, four amino acids, Asp-Glu-Val-Gly (DEVG), were selected as a negative control. Subsequently, the recombinant gene was cloned into the N and C terminal end of the split intein, and named as pFluc-DEVD and pFluc-DEVG. Then the plasmids were transfected into cells and renilla luciferase was co-transfected in each sample as an internal control for transfection efficiency. Then the apoptosis level was detected by the double luciferase reporter gene and the Western blotting analysis. The results showed that when apoptosis occurred, the content of firefly luciferase expressed in the pFluc-DEVD plasmid transfected group was about 3 times higher than pFluc-DEVG plasmid transfected group. Furthermore, Western blotting detection indicated that the Fluc level was significantly increased in pFluc-DEVD transfected group when pre-treated by apoptosis stimulants. The activation degree of Caspase-3 was closely related to the expression of Fluc, and had a significant statistical difference. These results confirmed that firefly luciferase protein expressed by pFluc-DEVD plasmid can be cleaved by the intracellular Caspase-3 enzyme, and this plasmid can accurately reflect the cell apoptosis level, which provides a useful method for quantitative detection of apoptosis.
Apoptosis ; Genes, Reporter ; Luciferases, Firefly ; Transfection

Objective To investigate the clinical characteristics and treatment strategies of arachnoid cyst combined with chronic subdural hematoma.Methods Seventeen patients with arachnoid cyst combined with chronic subdural hematoma,who received treatment in our hospital from February 2008 to March 2016,were chosen in our study;13 of them were performed burr hole irrigation and drainage,2 received craniotomy for removing hematoma and cystectomy,and 2 received endoscopic cyst excision and hematoma removal surgery.A retrospectively analysis of clinical characteristics and treatment strategies of these patients was performed.Results Follow up for 0.5-2 years was performed in the 13 patients performed burr hole irrigation and drainage,and no hematoma relapse was noted;one was found rebleeding in the cyst associated with a small amount of subdural hematoma on the 5th d of surgery,and the subdural hematoma was absorbed during the follow-up observation;one was found that the cyst and subdural hematoma both disappeared in the postoperative check,while the cyst reappeared 40 days after surgery,and during the follow-up observation,the cyst was not enlarged and the chronic subdural hematoma was not relapsed.Two patients recovered after receiving the craniotomy for removing hematoma and cystectomy;two patients recovered after receiving the endoscopic cyst excision andhematoma removal surgery.Conclusion For patients with arachnoid cyst associated with chronic subdural haematoma,burr-hole irrigation and drainage are generally selected;for patients with reappeared cyst or hematoma,there is no need to burr hole again or perform the surgery aiming at the cyst in a haste,and follow-up observation is suggested;for patients whose symptoms in the cyst have appeared before hemorrhage or whose hemorrhage appeared recurrently in the cyst,surgery of hematoma removal and cyst excision can be chosen at the same time.

Objective To investigate the effect of inhibiting tumor suppressor gene phosphatase and tensin homology deleted on chromosome ten (PTEN) on growth of neurons cultured in inhibitive microenvironment.Methods Primary cortical neurons were separated from fetal rats and cultured with adult rat brain-derived myelin membrane proteins as inhibitory substrate.Cells were divided into 3 groups:normal control group,inhibitory control group (myelin membrane proteins as inhibitory substrate) and bpv treatment group (200 nmol/L bpv+myelin membrane proteins).Real-time quantitative-PCR was performed to detect the expressions of PTEN and mammalian target ofrapamycin (mTOR) genes,and immunofluorescence staining was used to observe the growth ofneurites 24,48 and 72 h,and 5 d after inoculation.Results Immunofluorescence staining revealed that significant differences of the growth of neurites at the four observation time points existed among the 3 groups (P＜0.05); during the early days of culture (24,48 and 72 h),cells in normal control group had the longest neurite (43.5±12.2,94.9±23.6 and 154.3±35.4 μm),those in the bpv treatment group enjoyed the second prize (28.8±10.2,75.4±22.5 and 136.8±40.8 μm) and those in the inhibitory control group was obviously inhibited (17.4±5.8,46.3± 13.7 and 78.4 ±29.8 μm).But the difference on the 5th d of cultivation changed into bpv treatment group＞normal control group＞inhibitory control group with the neurite length of (225.9±50.6),(218.4±58.1) and (173.6±48.7) μm,respectively (P＜0.05).PCR revealed no significant difference in expressions of PTEN and mTOR genes of the three groups between each two time points (P＞0.05); while at the same time point,significantly increased expressions ofmTOR and PTEN in the bpv treatment group were noted as compared with those in the normal control group and inhibitory control group (P＜0.05).Conclusions Growth of neurons in inhibitive microenvironment is improved on condition that the tumor suppressor gene PTEN is inhibited,which is probably related to the activation of PI3K/AKT/MTOR pathway.

Objective To introduce the clinical design of the portable and moveable laser light for simulation immobilization.Methods According to the theory of the laser immobilization system,the setting is designed by using current material,such as a ruler of ?-shape and pillar-shape laser light etc.After installation and adjustment,it is used in clinic.Results Patient can be immobilized by CT and this setting.Being simple to operate and convenient to use,this setting can act for the expensive laser position system.Conclusion The material and creation of this setting are rougher,so it needs improvement to insure the accuracy of position.

Objective To discuss the three-dimensional conformal radiotherapy(3D-CRT) plan for pancreatic tumors.Methods The 3D-CRT plan of 30 cases of primary pancreatic cancer were designed according to the request of the doctors for PTV and the restricted receiving dose of the peripheral normal sensitive organs.The best plan was chosen referring the dose-volume histogram.Results By using ICRU dose reference point(the central point of target)as theunitary dose,95% dose line could be surrounded above 95% of the PTV.Conclusion The focus in different localization can be treated with different design in treating the pancreatic tumors.The reaction to radiotherapy is directly influenced by the volume and dosage of radiation to the normal hepatic tissues.

It is not only a radiotherapy reform but a practical and effective treatment that the moulding of low-melting-point lead is used to supplement conventional and normal lead block.Based on clinical practice,the significance and implementation of integrated block are discussed in this paper.

Objective To apply lead block technology to the interdiction of X-ray destroy from linear accelerator to normal tissues and sensitive organs so as to treat large fields and irregular malignant lymphoma.Methods The steps were as follows:(1)immobilizing patients;(2)simulator photographing;(3)sketching targets;(4)tailoring lead block;(5)validating lead block.Results Problems of large fields and more targets were solved in radiotherapy for malignant lymphoma patients.Conclusion Based on clinical practice,new methods was found for realizing large irregular radiation fields.

Objective To investigate and carry out the function of the lead block validating instrument in the radiotherapy. Methods Based on clinical practice and the principle comparative steady STD and SAD of linear accelerator, the function of lead block validating instrument was developed in reason. Results The working quality and efficiency are greatly improved. Conclusion Not only the criterion extent of founded lead block and the proof of every kinds spume blocks in three-dimensional conformal radiotherapy and integer block radiotherapy and electron radiotherapy, but also the shadow tray centers and center reticle can be validated by the lead block validating instrument.