Local icariin application enhanced periodontal tissue regeneration and relieved local inflammation in a minipig model of periodontitis

Periodontitis is an inflammatory autoimmune disease. Treatment should alleviate inflammation, regulate the immune reaction and promote periodontal tissue regeneration. Icariin is the main active ingredient of Epimedii Folium, and it is a promising compound for the enhancement of mesenchymal stem cell function, promotion of bone formation, inhibition of bone resorption, alleviation of inflammation and regulation of immunity. The study investigated the effect of icariin on periodontal tissue regeneration in a minipig model of periodontitis. The minipig model of periodontitis was established. Icariin was injected locally. The periodontal clinical assessment index, a computed tomography(CT) scan, histopathology and enzyme-linked immune sorbent assay(ELISA)were used to evaluate the effects of icariin. Quantitative analysis results 12 weeks post-injection demonstrated that probing depth,gingival recession, attachment loss and alveolar bone regeneration values were(3.72 ± 1.18) mm vs.(6.56 ± 1.47) mm,(1.67 ± 0.59)mm vs.(2.38 ± 0.61) mm,(5.56 ± 1.29) mm vs.(8.61 ± 1.72) mm, and(25.65 ± 5.13) mm3 vs.(9.48 ± 1.78) mm3 in the icariin group and0.9% NaCl group, respectively. The clinical assessment, CT scan, and histopathology results demonstrated significant enhancement of periodontal tissue regeneration in the icariin group compared to the 0.9% NaCl group. The ELISA results suggested that the concentration of interleukin-1 beta(IL-1β) in the icariin group was downregulated compared to the 0.9% NaCl group, which indicates that local injection of icariin relieved local inflammation in a minipig model of periodontitis. Local injection of icariin promoted periodontal tissue regeneration and exerted anti-inflammatory and immunomodulatory function. These results support the application of icariin for the clinical treatment of periodontitis.

ENDOCOCHLEAR POTENTIAL AND POTASSIUM CONCENTRATION RECORDING IN MINIPIG COCHLEA

Objective To record the endocochlear potential (EP) and calculate potassium concentration [K+] in Minipig cochleae. Methods We used multi-barreled electrodes to measure the EP and the potential, [K + ]. EP and potassium electrode recording were made in 9 cochleae from 5 minipigs to get normal EP values. Results The average EP value in the cochlea from the minipigs was 77.3 ± 14 mV. The average [K+] for the minipigs was 147.1 ± 13 mM. Conclusions The EP and potential, [K + ] in minipigs are lower than in the human and rodents. This may be the reason why porcine ABR thresholds are slightly higher than humans and rodents.

Transcript variants and expression profiles analysis of Mitf gene in minipigs

Object： To identify transcript variants and expression patterns of porcine Mitf. Materials and methods： A pairwise BLAST search at NCBI database was performed to deduce the structure of porcine Mitf gene. Subsequently, 5＇ RACE and fluorescent quantitative RT-PCR were used to analyze the expression pattern of porcine Mitf in different tissues. Results： Four transcript variants of porcine Mitf, MITF-A, MITF-H, MITF-M and MITF-SUS were identified, all sharing high homology with those in humans, except Mitf-SUS.Conclusion： The sequence of porcine Mitf appear highly homologous to human MITF. However, only 4 transcript variants of porcine Mitf were identified in these minipigs, less than the 9 transcript variants in human MITF.

In vivo Digestive Stability of Soybean β-conglycinin β-subunit in WZS Minipigs

By now, the digestive stability experiments provided by most authoritative organizations are in vitro tests. Evaluating the protein digestive stability with in vivo models should be more objective. The present study aimed to verify the in vivo digestibility of soybean β-conglycinin β-subunit in Wuzhishan （WZS） minipigs. Three minipigs were surgically fitted with O-stomach and T-ileum cannulae and fed with soybean meals. According to SDS-PAGE, the 50 kD fraction of soybean β-conglycinin β-subunit persisted in the gastric fluid until 6 h after feeding, which was detected at 3 h and clearly visible at 4-6 h in the intestinal fluid. Western blot with anti-β-conglycinin β-subunit McAb confirmed it.

Effects of partial portal vein arterialization on liver regeneration after hepatectomy in minipigs with obstructive jaundice

Background Hilar cholangiocarcinoma is a malignant tumor that is difficult to cure. The aim of this study was to observe the effects of flow-controlled partial portal vein arterializations （PPVA） on liver regeneration after hepatectomy in minipigs with chronic obstructive jaundice. Methods Eight minipigs were made into chronic obstructive jaundice models. United semi-hepatectomy, which imitates extended radical surgery for treatment of hilar cholangiocarcinoma, was then performed. The eight minipigs were randomly divided into groups A and B （n=4 minipigs each）. PPVA was performed in Group A but not in Group B. The effects of flow-controlled PPVA on live regeneration after hepatectomy were observed for 30 days after hepatectomy. Results The portal vein PO2 at the immediate time point and on postoperative day 30 was higher in Group A （（47.33±2.43） and （48.50±4.44） mmHg） than in Group B （（35.38±4.06） and （35.55±2.55） mmHg respectively, all P 〈0.01）. The mitotic index of liver cells on postoperative days 14 and 21 was higher in Group A （12.55%±2.85% and 15.25%±1.99% respectively） than in Group B （6.85%±2.10% and 11.88%±1.15% respectively, all P 〈0.05）. The regeneration rate of residual liver on postoperative days 14 and 21 was higher in Group A （24.56%±6.15% and 70.63%±9.83% respectively） than in Group B （11.96%±5.43% and 44.92%±7.42% respectively, P 〈0.05 and P 〈0.01 respectively）.Conclusion Flow-controlled PPVA can promote liver regeneration after hepatectomy and prevent liver failure in minipigs with chronic obstructive jaundice.

Expressed sequence tags analysis of a liver tissue cDNA library from a highly inbred minipig line

Background Porcine liver performing efficient physiological functions in the human body is prerequisite for successful liver xenotransplantation. However, the protein differences between pig and human remain largely unexplored. Therefore we investigated the liver expression profile of a highly inbred minipig line. Methods A cDNA library was constructed from liver tissue of an inbred Banna minipig. Two hundred randomly selected clones were sequenced then analysed by BLAST programme. Results Alignments of the sequences showed 44% encoded previously known porcine genes. Among the 56% unknown genes, sequences of 72 clones had high similarities with known genes of other species and the similarities to human were mostly above 0.80. The other 40 clones showing no similarity to genes in National Centre for Biotechnology Information are newly discovered, expressed sequence tags specific to liver of inbred Banna minipig. Twenty-two of the 200 clones had full length encoding regions, 38 complete 5＇ terminal sequences and 140 complete 3＇ terminal sequences.Conclusion These newly discovered expression sequences may be an important resource for research involving physiological characteristics and medical usage of inbred pigs and contribute to matching studies in xenotransplantation.

水凝胶创伤修复研究中应用的小型猪模型

背景:小型猪因皮肤组织结构、心血管系统等与人高度相似,所以常被应用于皮肤损伤、血管创伤与美容医学等研究领域。水凝胶作为创伤修复敷料拥有较强的保水性和黏附性等多种优异理化性质,能够为伤口提供隔离湿润和药物释放作用。目的:归纳和总结水凝胶治疗小型猪不同实验用途创伤修复模型的应用进展,揭示各类小型猪创伤修复模型的发展现状,分析现阶段小型猪创伤修复模型的不足。方法:查阅Web of Science数据库及中国知网从数据库建立至2023年发表的相关文献,设置中文检索词为“小猪,小型猪,微型猪,迷你猪;凝胶,水凝胶;创伤,损伤,伤口,创口”等,英文检索词为“miniature Swine,miniature pig,minipig;gel,hydrogel;injury,wound,lesion,incision”等。共检索到438篇中、英文文献,通过纳入和排除标准共纳入59篇文献进行探讨。结果与结论:①目前用于创伤修复的模型主要有大型动物物种(犬和猪)、兔子及啮齿类动物(大鼠和小鼠),由于小型猪皮肤结构与人类较相似,因此小型猪是创伤修复理想的动物模型。②在体外皮肤损伤模型中,皮肤缺损模型是基础的伤口模型,按伤口缺损深度可分为全层皮肤缺损模型和中厚皮肤缺损模型;烧伤伤口模型和感染伤口模型是在皮肤缺损模型的基础上施加热金属烫伤和细菌培养的多维度模型,具有安全系数高、操作难度低的优点。③在体内创伤修复模型中,小型猪可用作更符合临床疾病病理状态的食管环切模型;在胃穿孔、血管止血模型中,小型猪可观地展示出水凝胶较强的脏器黏附性、止血性能及促进组织再生效应。④在猪的特定部位也有相应模式用途:猪耳通常用来评价水凝胶的药物缓释效果;猪细胞蛋白和猪皮胶原多制备组织来源的复合水凝胶。

Using Huntingtin Knock-In Minipigs to Fill the Gap Between Mouse Models and Patients with Huntington's Disease

Huntington＇s disease （HD） is an inherited autosomal dominant neurodegenerative disease characterized by pro- gressive motor deficits, cognitive decline, and psychiatric symptoms. It is caused by a pathological expansion of CAG trinucleotide repeats in exon 1 of the HD gene, resulting in the translation of a mutant form of huntingtin protein （mutant Htt） with an expanded polyglutamine domain in the N-terminal region [1 ]. Despite great progress in understanding the pathogenesis of HD using multiple mouse models, the exact mechanisms by which mutant Htt induces neuronal dysfunction and death are still not completely clear, and there is no curative treatment for this disease. An important reason is that the mouse, which is the most widely used animal model in HD research, differs from the human in many aspects, including the physiology, drug metabolism, blood-brain barrier, life span, brain volume, and neuroanatomical organization [2]. Thus, it is necessary to establish HD models with higher species than rodents, such as the dog, pig, and non- human primate, so as to bridge the gap between preclinical mouse models and clinical studies.

铁过载致心肌纤维化小型猪模型的建立

目的建立铁过载致心肌纤维化的小型猪模型,探讨铁过载致心肌纤维化的作用机制/病理生理特征。方法选取巴马小型猪18只,随机分为实验组15只、对照组3只。每隔7 d经肌内注射右旋糖酐铁溶液1次,首次剂量为400 mg/kg,其余剂量为200 mg/kg,对照组注射与实验组相同剂量生理盐水,于注射铁剂6 d后随机处死1~2只巴马小型猪;取部分心脏组织切片进行心肌胶原容积分数(CVF)测定和心肌铁半定量测定,剩余心脏组织用于测量心脏铁浓度(CIC)。结果随注射次数增多,实验组巴马猪CIC呈上升趋势,心肌胶原纤维及铁颗粒占视野面积比例增多。注射铁总量与CIC、心肌铁病理半定量、CVF之间高度正相关(r=0.957,r=0.971,r=0.957,P<0.001),CIC与CVF之间高度正相关(r=0.924,P<0.001),心肌铁半定量与CIC、CVF之间高度相关(r=0.973,P<0.001;r=0.944,P<0.001)。结论通过肌内注射首次剂量为400 mg/kg,后续剂量为200 mg/kg的右旋糖酐铁,共15次,可以建立心肌纤维化小型猪模型,心肌铁颗粒沉积越多提示心肌纤维化程度越严重。

小型猪在中医动物模型中的应用研究进展

中医动物模型是开展中医药研究的实验基础和必备工具。小型猪具有遗传特性稳定、体型小、易繁殖、性格温顺、易饲养、生理生化水平和疾病病理表现与人类相似等特点,是构建实验动物模型的理想选择,在医药领域具有广阔的应用前景及研究价值。目前已有学者利用小型猪构建了虚寒证、血瘀证、实热证、脾气虚证等单纯证候模型;动脉粥样硬化脾虚痰浊证和冠心病痰瘀互结证等中医病证结合模型,均取得较好成果。但目前小型猪在中医动物模型中的应用尚不广泛,值得进一步探索。

小型猪动脉粥样硬化斑块稳定性模型研究

目前已有的动物模型在研究动脉粥样硬化斑块破裂、破裂的可控性及量化研究方面均不能满足研究的需要.为了建立类似于人类动脉粥样硬化病变的斑块模型,体外研究斑块稳定性,应用传统的高脂高胆固醇膳食诱导建立了小型猪动脉粥样硬化模型,并从血脂水平和斑块病理形态学特征方面加以了证实.该模型中斑块与人类成熟斑块的高度相似性使其成为研究斑块稳定性和斑块破裂的较好模型.从量化比较这一出发点着手,建立了一个体外可控可量化诱导斑块破裂模型,方法简单易行,是一个较好的量化研究斑块破裂和破裂相关因素间关系的实验模型.

转SCK基因大米的营养学评价—小型猪比较喂养研究

目的:通过小型猪喂养实验,分析和评价转豇豆胰蛋白酶抑制剂基因大米(转SCK基因大米)的营养价值,并比较与亲本大米之间的差异。方法:选择断乳期雄性中国实验用小型猪15只,初始体重(6.22±0.42)kg,按体重和窝别随机等分成3组,分别为普通饲料组(NG),仅饲喂市售猪饲料;对照大米组(CG),饲喂70%亲本大米+基础饲料;转基因大米组(GG),饲喂70%转SCK基因大米+基础饲料。基础饲料包括12%小麦麸,8%豆粕,6%酪蛋白和4%的矿物质和维生素混合物。喂养62d,观察和比较三组动物生长发育、摄食量、血生化指标、骨骼发育、组织病理等方面的差异。结果:GG组动物与CG组的一般营养状况、血常规和血生化值、体格发育、骨骼与肌肉发育和脏器发育及病理方面均未见明显差异,其营养作用基本等同,且两个大米组动物的体重增长值均高于NG组(P<0.05)。GG和CG组动物的食物利用率相近,但均高于NG组(P<0.05)。结论:转SCK基因大米与对照大米对小型猪的生长发育的影响基本等同,未发现转SCK基因大米造成任何可观察到的非预期不良结果。

不同海拔地区两种小型猪血液生理生化指标比较

本试验旨在比较高原蕨麻小型猪与平原巴马小型猪的血液生理生化指标差异,为高原蕨麻小型猪的实验动物化提供理论基础。试验分别在高原地区和西安市采集高原蕨麻小型猪与巴马小型猪血样,检测10项血液生理指标和14项血液生化指标,并对结果进行统计分析。结果显示,血液生理指标中高原蕨麻小型猪的红细胞计数(RBC)、血红蛋白(HGB)、红细胞压积(HCT)、红细胞平均血红蛋白含量(MCH)、红细胞平均血红蛋白浓度(MCMH)均极显著高于巴马小型猪(P<0.01),血小板(PLT)、红细胞分布列宽变异系数(RDW-CV)和红细胞分布宽度标准差(RDW-SD)与巴马小型猪相比差异显著(P<0.05);血液生化指标中高原蕨麻小型猪γ-谷氨酰基转移酶(GGT)、葡萄糖(GLU)、肌酐(CRE)均极显著高于巴马小型猪(P<0.01),其余指标均无显著差异(P>0.05)。结果表明,高原蕨麻小型猪具有低氧适应的生理特征,与巴马小型猪相比有独特的高原适应性,具有开发成为高原疾病动物模型的潜力。

小型猪心肌缺血血瘀证动物模型的复制方法

目的:建立中华小型猪心肌缺血血瘀证的模型。方法:在小型猪左冠状动脉前降支第一间隔支起始部远端放置Ameroid缩窄环,使Ameroid缩窄环内层缓慢膨胀压迫冠状动脉,造成慢性心肌缺血模型。术后,对动物的行为改变、一般情况及血液流变学指标等进行动态观察。结果:手术后4周冠状动脉造影显示心肌缺血小型猪冠状动脉狭窄率大干75%,其行为及一般情况、舌象和血液流变学指标等与术前和同期假手术组相比,差异有统计学意义(P<0.05)。结论:术后4周,该模型之证候属于中医血瘀证范畴,而且其发病过程与人类慢性心肌缺血的病理过程相似,适用于心肌缺血血瘀证的研究。

跑步运动联合高脂单笼喂饲建立巴马小型猪高脂血症脾虚痰浊证动物模型

目的:通过跑步运动联合高脂单笼喂饲建立巴马小型猪高脂血症脾虚痰浊证动物模型。方法:选取6月龄雄性去势普通级小型猪47只,以随机数字表法随机分为对照组和模型组,对照组5只,模型组42只。模型组施以每日跑步训练联合高脂喂饲干预,对照组予普通饲料喂食,共喂饲24周。对巴马小型猪等待喂食行为、皮毛光泽、口色、粪便性状、划痕、生命体征、形体指标、血脂和行为学指标进行观测。结果:跑步运动联合高脂喂饲可使巴马小型猪出现高脂血症脾虚痰浊证表现:(1)模型组血清TC(14.89±5.56)mmol/L高于对照组(3.88±1.89)mmol/L,TG(1.00±0.36)mmol/L高于对照组(0.35±0.09)mmol/L,HDL-C(1.97±0.61)mmol/L高于对照组(0.77±0.17)mmol/L,LDL-C(8.00±3.17)mmol/L高于对照组(1.09±0.25)mmol/L,ox-LDL(8.84±0.86)μg/L高于对照组(8.04±0.25)μg/L,差异均有统计学意义(P<0.01)。(2)模型组更趋于倦怠、懒惰,表现为咬物品、摆尾频次、闻物体、站立、走持续时间等低于对照组,闻同伴、卧、坐持续时间高于对照组(P<0.01)。(3)模型组等待喂食行为、粪便性状、划痕、口色评分大于对照组(P<0.05)。(4)模型组体质量、腹围、臀宽大于对照组(P<0.01)。(5)模型组心率大于对照组,呼吸频率小于对照组(P<0.01)。结论:跑步运动联合高脂单笼喂饲能成功建立巴马小型猪高脂血症脾虚痰浊证动物模型,为巴马小型猪进一步应用于中医药研究中提供有益的参考。

西双版纳小耳猪群体遗传结构的RFLP分析

目的 对西双版纳小耳猪的群体遗传结构进行RFLP分析。方法 采用ECL核酸直接标记和检测系统 ,以HRP标记浕 -珠蛋白 - 3’HVR多基因座小卫星探针 ,对西双版纳小耳猪的HinfⅠ或HaeⅢ酶切片段进行Southern杂交 ,以化学发光法进行检测 ,获得了清晰可辨的、具有高度多态性的DNA指纹图谱。结果 2kb以上的谱带数在 6～ 1 5条之间 ,HinfⅠ酶切产生的图带数低于HaeⅢ。JB亚系内 80 5、 80 7家系和JS亚系内 1 1 1、 1 2 1、1 2 1、 1 51家系不同个体间的相似系数分别为 0 94± 0 0 9、 0 85± 0 0 8、 0 84± 1 7、 0 78±1 6、 0 83± 0 42、 0 87± 0 0 7,显著高于各家系间的相似系数 ,JB亚系内的 80 5家系和JS亚系内的 1 51家系不同个体间相似系数较大 ,分别为 0 94和 0 87。结论 西双版纳小耳猪近交程度较高 ,个体间差异较小 。

巴马小型猪在医学研究中的应用进展

广西巴马小型猪作为国内小型猪主要品种之一,具有遗传性稳定、多产、体重较小、体表多覆白毛等特征,组织器官及生化指标与人类相似,已越来越广泛地应用于医学研究领域:猪的心脏解剖与生理特点与人类高度相似,已被广泛应用于心血管系统研究中,在我国,巴马小型猪被用来构建心肌缺血、卵圆孔未闭等心血管疾病模型;猪具有杂食性及与人相似的脂质代谢,可用于研究内分泌疾病,巴马小型猪已用于糖尿病动物模型建立及其遗传易感性、并发症的防治研究等;巴马小型猪的消化系统与人类相似,利用此特点已建立阻塞性慢性胰腺炎、结肠穿孔、胆肠吻合等消化系统疾病模型;巴马小型猪除头、尾外,体表覆以白毛,这一特点使其成为研究皮肤创伤、烧伤修复等的理想动物;小型猪的牙齿解剖结构与人类相似,口裂大,可作为口腔医学研究中的理想动物,巴马小型猪已用来建立牙髓坏死模型及对上颌扩弓方式的研究;类似人的解剖、生理、病理使其成为较为适合的异种移植供体。在中医药研究方面,已用巴马小型猪分别建立了肝脏、脾脏、股动、静脉出血及头颈恶性肿瘤放疗后的腮腺损伤动物模型以研究中药制剂的疗效及机制。

三个品系小型猪Ⅱ型糖尿病模型的比较研究

目的比较三个品系小型猪对高糖高脂饲料诱发Ⅱ型糖尿病模型的敏感性，为我国小型猪在Ⅱ型糖尿病研究中的应用提供基础资料。方法4～5月龄巴马小型猪、五指山小型猪和中国农大小型猪各6头，以4头中国农大小型猪为对照，同步开展高糖高脂饲料喂养试验，试验周期8个月。每月称1次体重，测定血糖、血清胰岛素含量，每2个月进行1次静脉葡萄糖耐量试验（IVGTT），测定脂质代谢相关指标。结果巴马小型猪和五指山小型猪对高糖、高脂饲料比较敏感，在饲喂5～6个月后，开始出现糖代谢异常，表现为显著肥胖、血清胰岛素升高和胰岛素敏感性下降。至实验后8个月，巴马小型猪和五指山小型猪各有1头发展为糖尿病，分别有3头和2头出现糖尿病前期症状。中国农大小型猪对高糖高脂饲料诱发糖尿病相对不敏感，实验期间空腹血糖水平、血清胰岛素含量、胰岛素敏感指数和IvGlTr一直无显著变化，仅在实验后7个月开始表现肥胖，8个月出现胰岛素敏感性减低。三个品系小型猪在高糖高脂饲喂后，血脂相关指标（TG，CHO，HDL-C，LDL-C）普遍升高，以巴马小型猪升高幅度最大。结论巴马小型猪和五指山小型猪对高糖、高脂饲料诱发Ⅱ型糖尿病比较敏感，品系内个体差异较大。中国农大小型猪相对不敏感。这一结果为进一步的Ⅱ型糖尿病相关基因分析和高敏感家系培育研究提供了基础资料。

氯胺酮与速眠新Ⅱ复合应用对小型猪麻醉的观察

为探讨氯胺酮与速眠新Ⅱ复合应用对实验小型猪的麻醉效果。将氯胺酮与速眠新Ⅱ等量混合,采用肌肉注射法(0.28mL/kg)对12头实验小型猪(11.33kg±0.83kg)进行复合麻醉,观察麻醉起效时间、麻醉持续时间及镇痛效果,并监测麻醉过程中体温、呼吸、心率等生理指标及有无呕吐、流涎等不良反应。小型猪成功麻醉12例,占100%,肌肉注射后(4.2±1.6)min进入麻醉状态,麻醉状态维持(87.3±15.4)min以上,麻醉期间肌肉松弛效果好,动物呼吸和心率平稳,无不良反应。结果表明,采用氯胺酮与速眠新Ⅱ复合麻醉效果好,麻醉过程平稳,生命体征稳定。氯胺酮与速眠新Ⅱ复合应用是较为简便、安全的小型猪麻醉方法,适合于耗时较长的手术或试验的开展。