Changes of Immunoreactive β-Endorphin in Plasma, Pituitary and Hypothalamus of Rats during Oxygen-Induced Convulsions

We observed for the first time the differences of immunoreactive β-endorphin(IR -β- EP) content in plasma, pituitary and hypothalamus of rats under various conditionsusing radioimmunoassay (RIA) and the effects of naloxone and β - endorphin (β- EP) antiserumon initial time of convulsions (ITC), severity of convulsions(SOC) and mortality on surface(MOS) of rats to hyperbaric oxygen(HBO). The results suggest thatβ- EP may partici-pate in the course of oxygen - induced convulsions and be one of endogenous convulsion - causingagents.

免疫球蛋白G4相关性唾液腺炎临床特点及诊疗进展

免疫球蛋白G4相关性唾液腺炎(IgG4-RS)是一种与IgG4密切相关的慢性自身免疫性疾病,其发病率低,病因和发病机制尚不明确,以唾液腺尤其是颌下腺无痛性持续性肿大为主要临床表现,多数患者血清IgG4升高,淋巴浆细胞浸润,后期全身多器官可继发病变,组织病理学是该病最可靠的诊断方法。临床上倾向于传统糖皮质激素作为首选用药,现阶段可尝试应用生物制剂进行靶向治疗。本文根据国内外文献,对IgG4-RS的病因、临床表现、诊断与鉴别诊断、治疗和预后的研究进展作一综述,以期为该病的临床诊疗和研究提供帮助。

细粒棘球绦虫膜联蛋白B5、B15和B25的原核表达和分泌特性分析

旨在探讨细粒棘球绦虫膜联蛋白B5、B15和B25的反应原性及其在中间宿主组织中的分泌特性,为进一步研究细粒棘球绦虫膜联蛋白与中间宿主的相互作用奠定基础。作者提取细粒棘球蚴原头蚴的总RNA并反转录为cDNA,以cDNA为模板PCR扩增获得EgANXB5、EgANXB15和EgANXB25基因全长编码序列,使用同源重组法构建pET32a-EgANXB5、pET32a-EgANXB15和pET32a-EgANXB25质粒并进行重组蛋白的原核表达,应用蛋白免疫印迹对上述重组蛋白进行鉴定,并采用免疫荧光染色试验分析EgANXB5、EgANXB15和EgANXB25在中间宿主组织中的分泌特性。结果显示:本研究成功克隆并表达出重组EgANXB5、EgANXB15和EgANXB25,并更正了EgANXB25的CDS序列(GenBank:OR245515.1)。蛋白免疫印迹结果显示,这3种蛋白均能够被感染细粒棘球绦虫的犬阳性血清和感染细粒棘球蚴的小鼠阳性血清所识别。同时,免疫荧光染色结果显示,在细粒棘球蚴包囊附近的肝实质组织中可以检测到EgANXB5、EgANXB15和EgANXB25的阳性信号。EgANXB5、EgANXB15和EgANXB25具有良好的反应原性,同时它们具有分泌进入包囊周围肝实质组织中的潜能,可能进一步参与细粒棘球蚴与宿主的相互作用。

酪蛋白糖巨肽对花生过敏原免疫反应性的影响

目的 探究酪蛋白糖巨肽(casein glycomacropeptides,CGMP)与花生过敏原相互作用并降低其免疫反应性的潜力。方法 通过蛋白-蛋白分子对接技术探讨CGMP与Ara h1、Ara h2是否有相互作用的潜力。进一步通过混合水浴加热制备CGMP与花生蛋白的混合溶液(mixed solution of casein glycomacropeptides and peanut proteins,MCGP),建立MCGP致敏、花生蛋白激发的BALB/c小鼠模型,研究MCGP对花生过敏反应的影响。最后使用圆二色谱法研究酪蛋白糖巨肽与Ara h1、Ara h2的相互作用力及对其结构的影响。结果 CGMP与Ara h1、Ara h2间存在次级键(盐桥、氢键、范德华力),部分作用于过敏原表位;与花生蛋白过敏组相比, MCGP致敏组血清中的花生蛋白特异性免疫球蛋白E (specific immunoglobulin E, sIgE)、sIgG1、sIgG2a含量显著下降,白介素-4 (interleukin-4, IL-4)、IL-5、组胺水平显著下降,转化生长因子-β (transforming growth factor-β, TGF-β)、γ干扰素(interferon-gamma, IFN-γ)水平显著升高, MCGP中Ara h2的α-螺旋与β-折叠的比例改变。结论 CGMP能够改变Ara h2的结构,遮蔽花生过敏原表位,抑制sIgE、sIgG结合Ara h1、Ara h2,降低部分花生过敏原的免疫反应性。

Effectiveness and safety of electroacupuncture analgesia in controlling intraoperative pain and hemodynamics during total thyroidectomy:A randomized controlled trial

Objective:To evaluate the effectiveness and safety of electroacupuncture in conjunction with additional medications in providing analgesia and stabilizing hemodynamic parameters during total thyroidectomy.Methods:This randomized controlled trial included 100 patients who underwent a total thyroidectomy between October 2022 and October 2023 at the Vietnam National Hospital of Acupuncture.The patients were randomized into two groups.The electroacupuncture analgesia(EA)group received EA stimulation at five acupuncture points:Hegu(LI 4),Neiguan(PC 6),Shuitu(ST 10),Quepen(ST 12),and Yifeng(SJ 17),while the control group received a bilateral superficial cervical plexus block.Primary outcomes included the level of analgesia and perioperative vital signs in both groups.Additionally,pain thresholds and serum b-endorphin levels were measured before and after electroacupuncture in the EA group.Results:Complete analgesia(Level A)was attained in 86%and 76%of the patients in the EA and control groups,respectively,with no significant difference between the two groups(P=1.00).In the EA group,the mean pain threshold after receiving EA doubled(648.7(77.4)g/s vs.305.3(45.3)g/s,P<.001),and the mean serum b-endorphin level increased by approximately 13.5 pg/mL(P<.001).All patients remained hemodynamically stable throughout the surgery.Conclusion:EA,in conjunction with additional medications that stimulate five acupuncture points,LI 4,PC 6,ST 10,ST 12,and SJ 17,was well tolerated and effectively maintained a suitable level of analgesia and hemodynamic stability during total thyroidectomy.

包埋青蛤免疫活性肽脂质体的制备及性质表征

为进一步提高青蛤免疫活性肽(Cyclina sinensis immunoceactive peptides,SCSP)的免疫调节功能,寻求更好的多肽递送方法,采用脂质体技术包埋SCSP,优化其制备工艺,并探究其理化性质和生物活性。比较了乙醇注入法、薄膜水合法、逆相蒸发法对包埋SCSP脂质体(SCS P-LIP)的平均粒径和包封率的影响;以平均粒径、包封率为评价指标,蛋黄卵磷脂质量、胆固醇与蛋黄卵磷脂质量比、SCSP与蛋黄卵磷脂质量比为变量,优化SCS P-LIP的制备工艺;考察了SCS P-LIP在4℃和25℃条件下贮藏7 d的稳定性;探究了SCS P-LIP对RAW 264.7细胞的免疫调节和细胞摄取效果。研究结果表明,薄膜水合法较适合SCS P-LIP的制备,优化制备条件为蛋黄卵磷脂质量40 mg、胆固醇与蛋黄卵磷脂质量比1∶4、SCSP与蛋黄卵磷脂质量比1∶4、探头超声12 min,在此条件下,SCS P-LIP的平均粒径为(107.500±2.301)nm,多分散指数为0.175±0.023,Zeta电位为(-29.800±0.577)mV,包封率为69.95%±1.24%。SCS P-LIP体系澄清透明,无特殊气味;透射电镜结果显示,SCS P-LIP呈球状,分布均匀;红外光谱结果显示,SCSP被包裹在脂质体极性部位;贮藏稳定性结果表明,SCS P-LIP更适合于4℃条件下避光保存;细胞实验结果表明,SCS P-LIP具有促RAW 264.7细胞增殖分化作用,对巨噬细胞吞噬功能的提升比SCSP更强,且具有较好的细胞摄取效果。研究结果表明,SCS P-LIP的制备工艺简单、性质稳定,有望为进一步开发具有免疫调节活性的多肽类功能食品提供技术参考。

重组粉尘螨变应原rDer f 27制备及其活性鉴定

目的:制备粉尘螨变应原第27组分(Der f 27)重组蛋白,鉴定其免疫活性。方法:构建pET-28a(+)-Der f 27质粒,转化至E.coli BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 27。IgE-ELISA和IgE-Western blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率,分别将变应性鼻炎患者PBMC、人支气管上皮细胞BESA-2B与rDer f 27共孵育24 h后测定细胞因子表达;生物信息学软件分析Der f 27的理化性质和结构。结果:成功构建pET-28a(+)-Der f 27质粒并转化至BL21(DE3)细胞,经IPTG诱导表达和纯化后,SDS-PAGE和Western blot在约48 kD处见特异性条带;IgE-ELISA和IgEWestern blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率分别为39.5%和45.5%;与对照组相比,变应性鼻炎患者PBMC与rDer f 27共孵育后,IL-6和IL-8表达升高(P<0.05);BESA-2B细胞与rDer f 27共孵育后,IL-10和TGF-β表达降低,IL-17A和IL-23A表达升高(P<0.05);生物信息学结果显示Der f 27属于丝氨酸蛋白酶抑制物家族,具有此家族通用结构及功能,二级结构主要为α螺旋(42.62%)和无规则卷曲(35.60%)。结论:成功制备了具有较好免疫反应性和免疫原性的重组变应原rDer f 27,并揭示其为变应性鼻炎的重要变应原之一。

应用纳米技术改进骨肉瘤诊治的研究现状

骨肉瘤作为一种恶性肿瘤,其早期诊断与有效治疗是医学界高度关注的焦点之一。随着纳米生物材料技术的发展,纳米技术在骨肉瘤的诊疗方面呈现了不可限量的潜力。本文以简略介绍骨肉瘤和纳米技术为基础,综合纳米材料的多种功能特性,详细探讨了纳米材料在骨肉瘤诊治中的研究现状,包括易修饰性、信号放大、高载药性、光热转换、免疫调节等方面。同时,展望了纳米技术在未来癌症检测和治疗领域的应用前景进,旨在为该领域未来的良好发展提供有益的参考。

Expression and immunoreactivity of an epitope of HCV in a foreign epitope presenting system

AIM: To construct and highly express an epitope of hepatitis C virus (HCV) in a foreign epitope presenting vector based on an insect virus, and to study the antigenicity of the epitope. METHODS: The HCV epitope sequence (amino acid residues 315 to 328: EGHRMAWDMMMNWS) of the El region was constructed at different positions of a foreign epitope presenting vector based on an insect virus, flock house virus (FHV) capsid protein encoding gene as a vector, and expressed in E. coli cells. Western blotting and ELISA were used to detect the immunoreactivity of these recombinant proteins. RESULTS: The gene encoding of the concerned B-cell epitope of HCV El envelope protein was expressed on FHV capsid carrier protein at positions I1 (aa 106), 12 (aa 153) and 13 (aa 305), respectively, on the surface of FHV capsid protein. The recombinant proteins in this system could be highly expressed in more than 40% of total cell protein of E. coli BL21. All the expressed recombinant proteins were in inclusion body form, and showed obvious immunoreactivity by Western blotting. Further purified recombinant proteins were detected by indirect ELISA as coating antigen respectively. All recombinant proteins could still show immunoreactivity. CONCLUSION: The epitope of HCV El envelope protein can be highly expressed in FHV carrier system as a chimeric protein with high immunoreactivity. This system has multiple entry sites conferring many possible conformations closer to the native one for a given sequence.

Expression and Immunoreactivity of a Human Group A Rotavirus Vp4

Rotavirus capsid protein Vp4 plays an important role in the virus adhering and entering the cells. In this study, a Vp4 gene cloned from a rotavirus strain TB-Chen was highly expressed in E.coli BL21 (DE3). The results of the Western blot showed that the protein possesses specific immuno-reactivities and can be specifically recognized by guinea pig antibodies against rotavirus strain SA11 or Wa. Some Vp4 dimers were formed during renaturation. These data obtained from this study provide a strong basis for further study on the structure and function of the Vp4.

Changes in beta-endorphin neuron numbers and serum hormone levels in the arcuate nucleus of ovariectomized rats undergoing treadmill exercise

BACKGROUND： The arcuate nucleus, when damaged in young rats, can lead to pathological changes in adults, such as osteoporosis. Ovariectomized rats suffer from osteoporosis at eight weeks following surgery and the number of β -endorphin immunoreactive neurons in the arcuate nucleus of the hypothalamus is significantly decreased. OBJECTIVE： To establish a rat model of osteoporosis using ovariectomy and to explore changes in the number of β-endorphin neurons and to correlate any such change with serum hormone levels in response to exercise or rest. DESIGN, TIME AND SETTING： The completely randomized block design, neural morphology study was performed at the Key Laboratory of Physiology, Guangdong Medical College, China between March 2004 and January 2005. MATERIALS： Sixteen healthy female rats were selected for ovariectomy. METHODS： Following model establishment, rats were assigned to either rest or exercise groups and each rat was housed individually. Rats in the exercise group underwent an exercise regimen using a treadmill. MAIN OUTCOME MEASURES： Eight weeks following exercise, radioimmunoassay was performed to detect serum growth hormone, estrogen and osteocalcin levels. Immunohistochemistry was used to measure changes in the number of β -endorphin neurons in the arcuate nucleus of the hypothalamus. Changes in bone metabolism were assessed using bone histomorphometry. RESULTS： In the exercise group, the β -endorphin immunoreactive neurons were high in number, darkly stained, and the nucleus was not obvious. In the rest group, the β -endorphin immunoreactive neurons were low in number and lightly stained. The number of β-endorphin immunoreactive neurons in the exercise group was higher compared with the rest group （t = 2.83, P 〈 0.05）. Estrogen levels were similar between the rest and exercise groups （P 〉 0.05）. Serum osteocalcin and growth hormone levels were significantly higher in the exercise group compared with the rest group （t = 2.78, 2.32, P 〈 0.05）. Compared with the rest group, the percentage of trabecular bone area, trabecular thickness, and trabecular number were significantly increased, but trabecular separation was significantly reduced （t=2.48, 2.57, 2.32, 3.06, P 〈 0.05） in the exercise group. In the exercise group, the trabeculae of the tibia were arranged regularly and were high in number. In the rest group, the trabeculae of the tibia were organized in a disorderly manner and were low in number, with many fat particles. CONCLUSION： Exercise promotes bone growth and delays osteoporosis by inducing an increase in the number of β-endorphin immunoreactive neurons in the arcuate nucleus of the hypothalamus and by increasing serum growth hormone and osteocalcin levels.

Trigeminal primary afferent terminals containing substance P－like immunoreactivity in the paratrigeminal nucleus

With the methods of transganglionic transport of horseradish peroxidase(HRP) combined with postembedding colloidal gold stain immunoelectron microscopy, the shape and synaptic relationship of the trigeminal primary afferent terminals containing substance

Effects of plasma β-endorphin on hemorrhagic shock in acute hypoxic rats

Forty rots were randomized into 2 groups and naloxone or saline were injected to therats after they were inflicted with hemorrhagic shock at sea level and at a simulated altitude of4000m respectively to observe the effects of naloxone on left ventricular systolic pressure(LVSP),left ventricular diastolic pressure(LVDP),the maximal changing rate of LVSP(dp/dt max),heartrate(HR),and survival time of the animals.Plasma β-endorphin(β-EP)was determined beforeand after hemorrhage to observe the relationship between β-EP and hemorrhagic shock.It wasfound that the circulatory parameters of hemorrhagic shock changed more markedly at high alti-tude than at sea level,naloxone could restore these parameters and prolong the survival time inboth the animals of the sea level and high altitude groups,and plasma β-EP level was elevatedafter hemorrhage especially in those animals at high altitude.These findings indicate:(1)Hemorrhagic shock at high altitude is usually accompanied with severe clinical manifestations,rapid progression,and high mortality.(2)β-EP seems to participate in the pathologicalmanifestafions of hemorrhagic shock at high altitude,and its depressive action on myocardialcontraction may be one of the factors inducing hemorrhagic shock.(3)Naloxone possesses defi-nite property to comet hemorrhagic shock at high altitude.

EFFECTS OF β-ENDORPHIN ON PHYTOHEMAGGLUTININ-INDUCED LYMPHOCYTE PROLIFERATION AND MOUSE PLAQUE-FORMING CELL RESPONSE VIA AN OPIOID RECEPTOR MECHANISM

The effects of opioid peptides on iminune responses were investigated. It was found that β-endorphin (β-END) can depress proliferative responses to PHA in rat splenocytes but enhance those in mice, and it could also inhibit the plaque-forming cell (PFC) response to sheep red blood cells when mouse splenocytes immunized in vivo were cultured in vitro with the peptide. The peptide antagonist naloxone was able to reverse β-END suppression of the PFC response. The data indicate that β-END suppresses antibody production or secretion via a specific opioidreceptor-mediated mechanism.

Expression of Endogenous Retrovirus ev/J gp85 Gene and Analysis of Its Immunoreactivity in Comparison with Exogenous Viral Protein

The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This report described expression of ev/J envelope gene gp85 derived from commercial meat-type chicken using the Invitrogen Bac-to-Bac baculovirus expression system. The antigenicity and immunoreactivity of the recombinant endogenous gp85 gene product (SU) were analyzed by indirect immunofluorescence, Western blot, indirect and blocking Enzyme-Linked ImmunoSorbent Assay (ELISA) using JE9 monoclonal antibody (MAb) against the envelope protein of ALV-J (ADOL-4817), positive mouse antiserum against the ev/J gp85 SU and sera from chicken naturally infected with ALV-J. The results showed that the ev/J gp85 SU can bind specifically to JE9 MAb and antiserum from chicken naturally infected with ALV-J, and the binding reactivity between exogenous ALV-J gp85 SU and natural positive chicken serum against exogenous ALV-J can be blocked by positive mouse serum against the ev/J gp85 SU. It is concluded that recombinant endogenous gp85 gene product (SU) has close immunological relatedness to the envelope protein of exogenous ALV-J (ADOL-4817 and IMC10200 strain).

LABELING McAb 3H11 AND ITS Fab FRAGMENT WITH ^（211）At AND THEIR IMMUNOREACTIVITIES AND INJURY EFFECTS ON HUMAN GASTRIC CANCER CELLS

An antigastric cancer monoclonal antibody, 3H11, and its Fab fragment, were labeled using p-(211At)-astatobenzoic acid (pAtBA) intermediate, in the yields of more than 30%. The results of in vitro experiments show that 211At-3H11 and 211At-3H11Fab are stable, and have specific immunoreactivities and cytotoxic effects to human gastric cancer cell M85. The cytotoxic effects are dependent on the concentration of 211At-3H11 or 211At-3H11 Fab and obviously stronger than that of Na211At.

Effects of Peripherally Acting Opioid Ligands on Central Opioid Receptors and <i>β</i>-Endorphin Release in Stressed Rats

Using the radioreceptor binding assay, μ-opioid receptor (MOR) affinity in the midbrain of stressed rats was higher than in naive controls. MOR density in the rat frontal cortex was reduced after stress. Intragastric administration of the MOR antagonist naloxone methiodide was followed by an increase in the number of MORs in the frontal cortex. However, the MOR agonist loperamide significantly decreased the density of MORs in the frontal cortex and midbrain of naive animals. Loperamide and naloxone methiodide were shown to prevent an increase in MOR affinity and a decrease in MOR density in the midbrain of rats after restraint stress. The restraint stress was accompanied by an increase in the release of β-endorphin (BE) in the ventral tegmental area (VTA) of control rats. After administration, loperamide slightly decreased the release of BE, naloxone methiodide significantly increased the release of BE in the cingulate cortex (CC) of untreated animals, while drugs had no effect on the release of BE in the VTA. The drugs significantly increased the extracellular level of BE in the CC of stressed animals. Loperamide abolished the increase in the stress-induced release of BE in the VTA. By contrast, naloxone methiodide significantly increased the release of BE in the VTA of stressed rats. Our data indicated that activation of peripheral MORs induces depression of the central part of the μ-opioid system, but suppression of peripheral MOR activity induces activation of the central μ-opioid system, the interaction of which can be modulated by stress.

Plasma β-Endorphin Levels in Women with Early Threatened Abortion before and after the Treatment of Integrated Chinese and Western Medicine

To observe plasm a β-endorphin (β-EP) and gonadotrophin releasing horm one (GnRH), hum an chorionic gonadotrophin (hCG), progesterone (P4) levelsin w om en w ith early threatened abortion and w ith a history of recurrent spontaneous abortions (RSA). Tw enty patientsw ith threatened abortion at7～8 w eeksof gestation w ere re- cruited, allof them had a history of 3 or m ore recurrentunexplained abortions. They w ere treated w ith psychologicalconsultation accompanied by traditionalChinese herbs. Blood samples w ere taken to m easure β-EP, GnRH, hCG and P4 levels by radioim - m unoassay (RIA). The treatm ents w ere continued till10～12 w eeks, blood w astaken during this period to compare changes in these peptides / horm ones. Tw enty norm al pregnantw om en at7～8 and 10～12 w eeksand 20 patientsw ith incompleteabortion at 10～12 w eeksw ererecruited for comparativestudies. Results: (1) In norm alpregnant w om en, plasm a β-EP, GnRH, hCGand P4 levelsat10～12 w eeksw ere significantly higher than thatat7～8 w eeks (P< 0.01). (2) In patients w ith threatened abortion and a history of RSA, plasm a β-EP levels at7～8 w eeks w ere significantly higher than those of norm al pregnantw om en (P< 0.01); on the contrary, plasm a GnRH, hCGand P4 levelsin these patientsw ere significantly low er than thosein norm alcases (P< 0.01). After treatm ent, 16 of the 20 patients succeeded in m aintaining their pregnancies, the levels of the four plasm a contents at10～12 w eeks w ere sim ilar to thosein norm alpregnantw om en (P> 0.05). (3) Plasm a β-EPlevelsin patientsw ith incomplete abortionsat10～12 w eeksw ere dram atically higher and GnRH, hCGand P4 levelsw ere low er than in norm alpregnantw om en (P< 0.01). β-EP m ightplay a role in the pathophysiology of spontaneousabortion.

The effect of low frequency electromagnetic field on plasma β-endorphin in human brain

The effect of electromagnetic field on plasma β endorphin in 30 patients with migraine were studied in the experiment. All subjects received a 20 minute repetitive transcranial magnetic stimulation (Frequency 10Hz, Average intensity 8mT) per time, and the total experiment lasted 20 times. Before and after the experiment, the EEG and plasma β endorphin were tested. The results show that the level of plasma β endorphin in patients blood increased significantly from (73.486±26.002)mg/ml to (116.934±67.592)mg/ml (p<0.01), and the EEG average magnitude of the migraine patients were improved obviously from 41.77μV to 47.42μV.

不同原核表达载体对非洲猪瘟病毒CD2v蛋白可溶性表达及免疫反应性比较

本研究旨在系统性地探究不同原核表达载体对非洲猪瘟病毒CD2v蛋白的可溶性表达差别,并利用临床非洲猪瘟病毒抗体阳性血清比较包涵体和可溶性CD2v蛋白的免疫反应性。利用5种原核表达载体pCold-TF、pET28a、pMAL-C6T、pGEX-4T-1、pET32a分别表达去除信号肽和跨膜区的非洲猪瘟病毒CD2v蛋白,利用镍-氨三乙酸(nickel-nitrilotriacetic acid,Ni-NTA)亲和层析法分别纯化源自pET28a和pCold-TF表达载体的包涵体和可溶性CD2v蛋白,并用间接酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)方法比较纯化蛋白的免疫反应性。结果显示,pCold-TF载体以表达可溶性蛋白为主,pMAL-C6T载体同时表达包涵体和可溶性蛋白,而其他载体主要表达包涵体蛋白。临床非洲猪瘟病毒抗体阳性血清检测数据显示,可溶性蛋白的免疫反应性显著优于包涵体蛋白(P<0.05)。pCold-TF载体的触发因子(trigger factor,TF)标签可促进CD2v蛋白的可溶性表达,其表达蛋白的免疫反应性优于包涵体蛋白。本研究结果为CD2v蛋白的免疫原性研究奠定了基础,亦为其他重要抗原的可溶性表达提供了思路。