RESEARCH OF THE SWITCHED RELUCTANCE STARTER/GENERATOR SYSTEMS

This paperfocuses on the sim ulation and testofthe sw itched reluctance starter/generator system s. Through the em ulationalanalysis ofthe initialstarting torque, the optim alturn-on section ofthe pow er sw itches is discovered. The fundam entaltheory ofthe generating operation is analyzed w ith the linearity m odel, and a new m ethod is presented based on voltage pulse w idth m odulation for the generating m ode control. Through the steady-state and optim ized em ulation ofthe outputpow er and system efficiency, the optim izationalcontrolapproach forthe generating m ode overa w ide speed range isintroduced. Atlast, the testofthe 3KW prototype system show sthatthe dynam icand static perform ance of this system isfine.

CRISPR/Cas9介导下UPS系统-E3泛素连接酶ITCH基因敲除细胞株的初步构建

目的:构建CRISPR/Cas9介导下泛素-蛋白酶体系统(ubiquitin proteasome system,UPS)-E3泛素连接酶(atrophin 1interacting protein 4,ITCH)基因敲除细胞株。方法:在人ITCH基因的PAS结构域内设计3个sgRNA,构建3个分别同时表达ITCH-sgRNA、Cas9和eGFP的三合一质粒。经测序序列正确的重组质粒用lipofectamine 3000转染HeLa细胞,24h后荧光倒置显微镜观察转染情况。结果:3个ITCHsgRNA寡核苷酸单链成功退火成为双链,经测序发现,设计的3个ITCH-sgRNA全部重组到了CRISPR/Cas9载体上。荧光检测发现在同时表达ITCH-sgRNA、Cas9和eGFP的三合一质粒转染的HeLa细胞内有绿色荧光。结论:成功构建了同时表达ITCH-sgRNA、Cas9和eGFP的三合一质粒,并将三合一重组质粒成功转入细胞中,为实现在HeLa细胞中完全敲除ITCH基因,从而为构建ITCH基因敲除稳定株奠定了基础。

Intradermal administration of formalin to the cheek induces itch as well as pain behaviors in rats

Objective · Formalin is a classic and most widely used algogenic substance, but its itchy effect is not clear. The present study aims to explore the hypothesis that formalin may induce itch as well as pain. Methods · Flinching, as well as licking and forelimb wiping of the site of injection were counted as pain responses, whereas biting and hind paw scratching of the cheek were counted as itchy responses. To discriminate formalin-induced sensations in rats, the irritant(saline as control) was injected, and then pain and itchy responses were recorded.Results · Intraplantar injection of formalin elicited biphasic behavior responses characterized as flinching, as well as biting or licking of the hind paw without significant gender differences. Following intradermal administration of formalin to the cheek, rats exhibited episodic forelimb wiping of the cheek, representative of pain. No gender difference was noticed for this type of behavior. In addition, episodes of hind paw scratches of the cheek, representative of pruritoceptive responses, also occurred. Interestingly, hind paw scratches appeared to be more pronounced in male than in female rats. Conclusion · Intradermal administration of formalin elicits pruritoceptive as well as nociceptive responses in rats.

Clinical Observation of Chuanbai Antipruritic Lotion"Water Film"Wet Compress Combined with Chloramphenicol Prednisone Liniment in the Treatment of Acute Eczema

[Objectives]To observe and analyze the clinical efficacy and possible mechanism of Chuanbai antipruritic lotion"water film"wet compress combined with chloramphenicol prednisone liniment in the treatment of acute eczema.[Methods]A total of 76 acute eczema cases admitted to Shiyan Taihe Hospital from January 2022 to March 2023 were divided into Western medicine treatment group and integrated traditional Chinese and Western medicine group.In the Western medicine treatment group,chloramphenicol and prednisone liniment was applied to the skin lesions and oral administration of cetirizine hydrochloride.The course of treatment in both groups was 2 weeks(w).The levels of interferon-γ(IFN-γ),interleukin-4(IL-4)and the ratio of IFN-γ/IL-4 in the peripheral blood of patients before and 2 weeks after treatment,as well as serum immunoglobulin E(IgE),anti-IgE antibody and histamine(HA)level,and with skin lesions disappearing time,skin oil,transepidermal water loss(TEWL),eczema area and severity index(EASI)score,total effective rate,degree of pruritus and traditional Chinese medicine quality of life scale(EPQOLS)score to evaluate the efficacy.[Results]Compared with the Western medicine treatment group at 2 w,the disappearance time of skin lesions in the integrated traditional Chinese and Western medicine group was shortened,TEWL,itching degree and EASI score,serum IgE and HA,and peripheral blood IL-4 levels were all decreased(P<0.05).IFN-γand IFN-γ/IL-4 ratio,anti-IgE antibody,EPQOLS score and total effective rate were all increased(P<0.05),and the difference was statistically significant.[Conclusions]Chuanbai antipruritic lotion"water film"wet compress combined with chloramphenicol prednisone liniment is an optimized and safe and efficient method for the treatment of acute eczema,which can quickly relieve the symptoms of inflammatory damage of eczema and restore the skin barrier function,dry dampness and relieve itch.Inhibition of the release of histamine active substances and regulation of immunity may be the main mechanisms.

ITCH对肺癌A549细胞增殖侵袭和凋亡的影响

目的探讨ITCH在肺癌细胞系中的表达以及ITCH沉默对肺癌A549细胞增殖、侵袭、凋亡的影响及其可能机制。方法采用Western blotting检测人正常肺上皮细胞系BEAS-2B和人肺癌细胞系A549、Calu3中ITCH蛋白的表达情况。采用PEI转染试剂分别向A549细胞转染si ITCH(si ITCH组)和si NC(si NC组),采用CCK-8法、Transwell实验和流式细胞术检测两组细胞增殖能力、侵袭能力和凋亡的变化,Western blotting检测两组ITCH、Bcl-2、Bax、Cyclin D1、MMP-2、MMP-9、β-catenin、E-cadherin蛋白表达。结果人肺正常上皮细胞系BEAS-2B中ITCH蛋白表达水平为0. 98±0. 043,显著低于Calu3细胞的1. 92±0. 073和A549细胞的2. 74±0. 151,差异均有统计学意义(P <0. 05)。si ITCH组96 h时细胞增殖倍数为1. 97±0. 021,显著低于si NC组的3. 72±0. 232(P <0. 05); si ITCH组细胞凋亡率为(43. 2±1. 52)%,显著高于si NC组的(2. 3±0. 32)%,差异有统计学意义(P <0. 05)。si ITCH组穿膜细胞数为(297. 2±22. 21)个,显著低于si NC组的(601. 2±3. 21)个,差异有统计学意义(P <0. 05)。与si NC组比较,si ITCH组Bcl-2、Cyclin D1、MMP-2、MMP-9、β-catenin表达水平降低,Bax、E-cadherin表达水平升高。结论 ITCH沉默通过调控MMP、Cyclin D1、Bcl-2/Bax等信号通路抑制肺癌细胞增殖、侵袭并诱导其凋亡。

乳腺癌组织中Cbl-b和Itch蛋白的表达变化及意义

目的观察Cbl-b和Itch蛋白在乳腺癌组织中的表达变化并探讨其意义。方法选择116例乳腺癌患者术后标本和30例乳腺癌患者癌旁正常乳腺组织(距癌组织边缘5 cm),应用免疫组化法检测Cbl-b和Itch蛋白的表达情况,并分析其与临床病理特征的关系。结果 116份乳腺癌组织中Cbl-b和Itch蛋白的阳性表达率分别为78.45%(91/116)、58.62%(68/116),30份癌旁正常乳腺组织中Cbl-b和Itch蛋白的阳性表达率分别为26.67%(8/30)、23.33%(7/30)。乳腺癌组织的表达均高于癌旁正常乳腺组织(P均<0.01)。Cbl-b、Itch蛋白表达与浸润深度、淋巴结转移及病理分期均呈正相关(r分别为0.172、0.119、0.117,0.145、0.134、0.138;P分别为0.014、0.022、0.017,0.028、0.027、0.025);Cbl-b和Itch蛋白阳性表达与患病年龄、病灶大小无相关性(P均>0.05)。结论乳腺癌组织中Cbl-b蛋白、Itch蛋白表达均升高,此二者参与乳腺癌的进展。

ITCH的功能及其与疾病的关系

ITCH又称AIP4(atrophin 1相互作用蛋白4),最初是在研究小鼠表皮颜色时被发现的。ITCH是一个单体蛋白,属于同源的HECT类型的泛素连接酶E3家族,参与体内多种蛋白质的泛素化降解过程;参与免疫反应;参与多种细胞过程,包括细胞循环、增殖和凋亡;在肿瘤的形成过程中发挥重要的作用。现就ITCH的功能及其与疾病的关系研究进展予以综述。

超声微泡介导Itch基因沉默增强T细胞对肺腺癌细胞LA795免疫杀伤作用

目的利用超声介导微泡破裂技术(UTMD)沉默T细胞Itch基因表达,观察转染T细胞对肺腺癌细胞LA795的体外免疫杀伤效率。方法磁珠分选纯化T细胞,构建靶向Itch基因的shRNA表达质粒,超声微泡介导转染48 h后,荧光显微镜和流式细胞仪评估细胞转染效率,Western blot检测Itch蛋白表达。转染72 h后,酶联免疫吸附法(ELISA)检测细胞上清液中IL-2和IFN-γ分泌水平,观察对比单纯T细胞、阴性对照T细胞及转染T细胞与小鼠肺腺癌细胞LA795共培养时肿瘤杀伤率。结果利用UTMD技术介导shRNA转染效率达到52.3%±3.8%,Itch蛋白表达能够有效被抑制。转染72 h后,超声微泡介导沉默Itch基因显著增加T细胞因子IL-2和IFN-γ分泌水平(P<0.05);与空白组或阴性对照组T细胞相比,在不同的靶效比水平(10∶1、20∶1、40∶1),转染T细胞杀瘤活性均明显增高(P<0.05)。结论利用UTMD技术介导shRNA转染能有效沉默Itch基因表达,促进T细胞免疫活性,增强T细胞对小鼠肺腺癌细胞LA795的体外免疫杀伤效率。

基于HepG2细胞胰岛素抵抗模型探讨miR-7-5p对Itch基因的靶向作用

目的:通过体外建立Hep G2细胞胰岛素抵抗模型,检测胰岛素抵抗状态下微小RNA-7-5p(miR-7-5p)及其预测靶基因Itch的差异表达,初步探讨miR-7-5p对Itch基因的靶向作用及其与胰岛素抵抗的关系。方法:采用适宜浓度的软脂酸诱导Hep G2细胞,建立体外胰岛素抵抗模型;基于生物信息学分析预测miR-7-5p可能作用的靶基因及其富集的相关信号通路;运用RT-q PCR和Western blot技术检测在胰岛素抵抗状态下miR-7-5p和Itch的表达变化。结果:0.25 mmol/L的软脂酸作用于Hep G2细胞24 h可诱导细胞产生胰岛素抵抗,RT-q PCR检测表明,与阴性对照组相比,胰岛素抵抗组的miR-7-5p表达下调(P<0.01)。生物信息学分析结果表明,miR-7-5p有相当数量的靶基因富集于泛素-蛋白酶体系统,其中E3泛素连接酶Itch基因是与胰岛素抵抗最为相关的靶基因;Western blot结果揭示,在胰岛素抵抗状态下,Hep G2细胞中Itch蛋白表达上调(P<0.01)。结论:miR-7-5p可能参与了胰岛素抵抗的病理生理过程,其机制可能通过靶向调控Itch基因的表达,进而直接或间接影响胰岛素信号通路的正常转导。

UPS-E3泛素连接酶ITCH与胰岛素抵抗的关系

糖尿病已成为严重的世界性公共卫生问题,预计至2045年全球糖尿病患者将超过6亿人[1]。胰岛素抵抗是2型糖尿病的重要表型,是指各种原因导致胰岛素促进葡萄糖摄取和利用率下降,机体代偿性地分泌过多胰岛素,产生高胰岛素血症。胰岛素是一种蛋白类激素,促进糖原、脂质和蛋白质合成等多种生物代谢过程,其作用的发挥依赖于胰岛素信号正常转导;胰岛素重要靶器官如肝脏、脂肪、肌肉、胰腺和脑中胰岛素信号通路阻断将导致胰岛素抵抗。

Expression of Notch pathway components (Numb, Itch, and Siah-1) in colorectal tumors: A clinicopathological study

BACKGROUND The role of the Notch pathway in carcinogenesis and tumor progression has been demonstrated in many organs,including the colon.Accordingly,studies aimed at developing therapies targeting this pathway in various cancers require the identification of several factors that may play a role in regulating Notch-1 expression.Although Numb,Itch,and seven in absentia homolog-1(Siah-1)have been shown to contribute to the regulation of Notch signaling,their role in colorectal carcinogenesis and tumor progression has not been fully elucidated to date.AIM To evaluate Numb,Itch,and Siah-1 expression in colorectal tumors to clarify their relationship with Notch-1 expression and their role in carcinogenesis and tumor behavior.METHODS Expression of Notch-1,Numb,Itch,and Siah-1 was investigated in 50 colorectal carcinomas,30 adenomas,and 20 healthy colonic tissues by immunohistochemistry and quantitative real-time polymerase chain reaction(PCR)analyses.RESULTS In contrast to Notch-1,which is expressed at higher levels in tumor tissues and adenomas,expression of Numb,Itch,and Siah-1 was stronger and more frequent in normal mucosa(P<0.01).There was a positive correlation between Notch-1 expression and high histological grade,the presence of lymph node metastasis,and advanced-stage tumors,whereas expression of Numb,Itch,and Siah-1 was absent or reduced in tumors with these clinicopathological parameters(P<0.05).In survival analysis,expression of Notch was related to poor prognosis but that of Numb,Itch,and Siah-1 correlated with improved survival(P<0.05).Multivariate analysis revealed Notch-1 expression and loss of Numb expression to be independent prognostic parameters together with lymph node metastasis(P<0.05).CONCLUSION Our findings support the role of Notch-1 in colorectal carcinoma and indicate that loss of Numb,Itch,and Siah-1 expression is associated with carcinogenesis.Our data also suggest that these three proteins might be involved in the Notch-1 pathway during colorectal carcinoma(CRC)progression and might play an essential role in approaches targeting Notch as novel molecular therapies for CRC.

Altered Expression of Differential Genes in Thoracic Spinal Cord Involved in Experimental Cholestatic Itch Mouse Model

The spinal origin of cholestatic itch in experimental obstructive jaundice mouse model remains poorly understood. In this study, the jaundice model was established by bile duct ligation （BDL） in mice, and differential gene expression patterns were analyzed in the lower thoracic spinal cord involved in cholestatic pruritus after BDL operation using high-throughput RNA sequencing. At 21st day after BDL, the expression levels of ENSRNOG00000060523, ENSRNOG00000058405 and ENSRNOG00000055193 mRNA were significantly up-regulated, and those of ENSRNOG00000042197, ENSRNOG00000008478, ENSRNOG00000019607, ENSRNOG00000020647, ENSRNOG00000046289, Gemin8, Serpina3n and Trim63 mRNA were significantly down-regulated in BDL group. The RNAseq data of selected mRNAs were validated by RT-qPCR. The expression levels of ENSRNOG00000042197, ENSRNOG00000008478, ENSRNOG00000019607, ENSRNOG00000020647, ENSRNOG00000046289 and Serpina3n mRNA were significantly down-regulated in BDL group. This study suggested that cholestatic pruritus in experimental obstructive jaundice mouse model is related with in the changes of gene expression profiles in spinal cord.

cir-ITCH在胃癌患者肿瘤组织中的表达及临床应用

目的检测胃癌患者组织中circ RNA ITCH(cir-ITCH)表达情况并评估其在胃癌诊断中的应用价值。方法采用实时荧光定量RT-PCR检测cir-ITCH在胃癌患者癌组织以及胃癌细胞系中的表达水平,分析cir-ITCH表达水平与胃癌患者临床病理参数的相关性。结果 cir-ITCH在胃癌细胞系SGC-7901(0.621±0.036)和MGC-803(0.118±0.003)中的表达水平明显低于胃黏膜上皮细胞GES-1(0.999±0.037),差异具有统计学意义(t分别为7.294和23.58,P均<0.01)。88例胃癌患者癌组织中cir-ITCH的表达水平[0.635(0.137,1.506)]较癌旁组织[3.042(0.808,7.972)]明显下调(Z=-5.976,P<0.01),且与TNM分期密切相关(r为0.137,P<0.05)。结论 cir-ITCH在胃癌组织中表达下调,并与TNM分期密切相关,有望成为潜在的胃癌诊断新分子标志物。

超声微泡介导沉默T淋巴细胞Itch基因对胃癌MFC细胞免疫杀伤作用的实验研究

目的利用超声介导微泡破裂技术(UTMD)沉默T淋巴细胞Itch基因表达,观察转染T细胞对胃癌MFC细胞的免疫杀伤效率。方法免疫磁珠分离T淋巴细胞,构建靶向Itch基因的sh RNA表达质粒,超声微泡介导转染48 h后,流式细胞仪分析T细胞转染成功率,Western blot测定T细胞Itch蛋白表达。转染24 h后,流式细胞仪检测T淋巴细胞活化早期标志CD69表达。转染72 h后,观察对比单纯T淋巴细胞、阴性对照T淋巴细胞及转染T淋巴细胞与小鼠胃癌MFC细胞共培养时肿瘤杀伤率。结果利用UTMD技术介导sh RNA转染效率达到51.9%,Itch蛋白表达能被有效抑制。转染24 h后,转染组T淋巴细胞早期活化标志CD69表达较其他组更高。转染72 h后,与阴性对照组和空白组相比,在不同的效靶比水平(10∶1、20∶1、40∶1),转染T淋巴细胞杀瘤活性均增强。结论利用UTMD技术介导sh RNA转染能有效沉默Itch基因表达,促进T淋巴细胞免疫活性,增强T淋巴细胞对胃癌MFC细胞的免疫杀伤效率。

The extract of Mucuna pruriens possesses histamine activity

Objective:To investigate the activity of the active principle in the spines of the seed pods of Mucuna pruriens using contraction of guinea pig ileum as index of pharmacological activity. Methods:The active principle was extracted with 0.0015 M NaCl.Muscle strips of guinea pig ileum were prepared and contractile responses were measured using a Kymograph.Two sets of experiment were conducted:（1）.The contraction of the ileum in presence of different concentrations of histamine,2-methylhistamine and the extract of Mucuna pruriens.（2）.The contractile response of the ileum in presence of different concentrations of the extract and antagonists including diphenhydramine,atropine and methysergide.Results:（1） The extract of Mucuna pruriens hair, 2- methylhistamine and histamine produced dose dependent contraction of guinea pig ileum (Extract ED50 = 13.0μg/mL,2-methylhistamine ED50=8.5μg/mL and histamine ED50=10.0μg/ mL).（2） Diphenhydramine,an H,antagonist competitively blocked the contractile response of the Mucuna pruriens extract（3） Coadminstration of the Mucuna pruriens extract either with different doses of antimuscarinic agent atropine or 5-hydroxytryptamine blocking agent methysergide did not alter the extract induced contractile response of the guinea pig ileum.Conclusion:These results demonstrate that the spines of Mucuna pruriens possess histamine activity which may contribute to its itching and painful irritation effects.

Topical use of ozone effectively alleviates the acute symptoms and quality of life of patients with moderate to severe bullous pemphigoid: a randomized controlled trial

Objective:To evaluate the adjuvant treatment of acute stage symptoms and quality of life of patients with bullous pemphigoid(BP)with ozone.Methods:We included 74 patients with moderate and severe BP hospitalized from July 2018 to December 2020.The participants were randomly divided into the control group and the ozone group,with 37 patients in each group(n=37).The control group used tap water to wash the whole body,and mupirocin ointment was used for the erosions.The ozone group used ozone water to wash the whole body,and the erosion place was coated with ozone oil.The bullous pemphigoid disease area index(BPDAI)score of BP disease was used to observe the clinical symptoms and skin lesions of the two groups before and on day 3,7,and 14 after treatment.Pruritus and sleep were observed before and on day 3,7,and 14 after treatment.The Dermatology Life Quality Index(DLQI)was used to observe the quality of life before and after treatment and the adverse reactions were recorded.Results:The total BPDAI score and erythema score in the ozone group were significantly different from those in the control group on day 3,7,and 14 after treatment(P<0.050).The scab drying time and erosion drying time of ozone group were shorter than that of control group(P<0.001).There were statistically significant differences between the ozone group and the control group in pruritus score and sleep score on day 3,7,and 14 after treatment(P<0.001).There was statistically significant difference in skin disease quality of life between the two groups 14 days after treatment(P<0.001).No significant adverse reactions and complications were observed in both groups.Conclusions:Ozone external use can rapidly improve the acute stage of edema erythema,erosion,and itching of BP,improve the quality of life of patients,is an effective treatment method,and wor thy of clinical promotion.

Vulvar Contact Dermatitis: Effect of a Treatment with a Anti-Inflammatory, Moisturizing and Healing Product on the Course of the Disease

Background: Vulvar contact dermatitis is inflammatory disorders of genital skin causing itch, burning, and plaques with overlying excoriation and linear fissures. Objective: To assess the effectiveness of a soft foam based on Tiab&#174 system (TF), complex of titanium micro crystals covalently bound with silver ions associated with hyaluronic acid, as a treatment of vulvar contact dermatitis. Method: 36 patients with the diagnosis of vulvar contact dermatitis received an 10-days treatment, during which the subjects applied TF, complex of titanium micro crystals covalently bound with silver ions associated with hyaluronic acid on the affected vulvar surfaces, used once a day. Clinical symptoms (itching and burning) and signs (vulvar surface disruption) were documented at baseline and after 1 months of follow-up. Results: There was a significant reduction of symptoms between the basal scores and the symptoms complaints 1 months of follow-up. Moreover a significant reduction of excoriations and linear fissures was observed. Conclusion: The present study shows that application of TF can be an effective agents for the reduction of symptom and sign of vulvar contact dermatitis, and the medical device can be used as a preventive agent in predisposing condition of vulvar discomfort.

AVERAGING IN OPTIMAL SWITCHING CONTROL PROBLEMS

The averaging in optimal switching control problems is considered under the following two cases: the switching cost does not depend on ε and the switching cost vanishes as ε tends to zero. The value function of the original fast problem converges locally uniformly to the value function of the averaged problem under both cases. The ways of averaging turn out to be different between both cases.

WW结构域及相关蛋白在肿瘤发生中的作用

WW结构域由35～40个氨基酸残基组成，存在两个高度保守的色氨酸残基，能特异地与富含脯氨酸的蛋白基序结合。WW结构域存在于多种蛋白中，广泛参与细胞内各种生化过程和信号通路。WW结构域及其参与构成的蛋白与包括癌症在内的多种人类疾病存在密切联系，成为疾病诊断、治疗和药物开发的新靶标。本论文中，我们综述了WW结构域及其参与构成的蛋白在肿瘤和癌症发生中的重要作用和研究进展。

Emerging role of Toll-like receptors in the control of pain and itch

Toll-like receptors （TLRs） are germline-encoded pattern-recognition receptors that initiate innate immune re- sponses by recognizing molecular structures shared by a wide range of pathogens, known as pathogen-associated molecular patterns （PAMPs）. After tissue injury or cellular stress, TLRs also detect endogenous ligands known as danger-associated molecular patterns （DAMPs）. TLRs are expressed in both non-neuronal and neuronal cell types in the central nervous system （CNS） and contribute to both infectious and non-infectious disorders in the CNS. Following tissue insult and nerve injury, TLRs （such as TLR2, TLR3, and TLR4） induce the activation of microglia and astrocytes and the production of the proinflammatory cytokines in the spinal cord, leading to the development and maintenance of inflammatory pain and neu- ropathic pain. In particular, primary sensory neurons, such as nociceptors, express TLRs （e.g., TLR4 and TLR7） to sense exogenous PAMPs and endogenous DAMPs released after tissue injury and cellular stress. These neuronal TLRs are new players in the processing of pain and itch by increasing the excitability of primary sensory neurons. Given the prevalence of chronic pain and itch and the suffering of affected people, insights into TLR signaling in the nervous system will open a new avenue for the management of clinical pain and itch.