目的:探讨跑台运动、二甲双胍及二者联合干预对2型糖尿病(type 2 diabetes mellitus,T2DM)小鼠主动脉ox-LDL、LOX-1蛋白表达、血管炎症反应与细胞凋亡的影响,评价防控血管损伤的干预手段.方法:清洁级雄性C57BL/6J小鼠随机分为正常组、T2DM造模组、跑台运动组、二甲双胍组和运动+二甲双胍组.运动组进行8周跑台运动,药物组进行8周二甲双胍干预.干预结束后,采用相应试剂盒检测脂质代谢指标;硝酸还原酶法检测血清NO水平;ELISA法检测血清胰岛素、ET-1、TNF-α、IL-1β水平;Western Blot法检测主动脉eNOS、ox-LDL、LOX-1、NF-κBp65、VCAM-1和Caspase-3蛋白表达.结果:1)高脂饲料联合STZ诱导的T2DM小鼠出现糖脂代谢紊乱,主动脉ox-LDL、LOX-1、NF-κBp65蛋白表达显著上调,但NO/ET-1未见显著性变化;继续高脂饲料喂养9周后,糖脂代谢进一步紊乱,机体处于炎症状态,NO/ET-1显著性降低,主动脉ox-LDL、LOX-1、NF-κBp65、VCAM-1和Caspase-3蛋白显著上调,eNOS蛋白表达显著下调.2)8周跑台运动、二甲双胍及二者联合干预均改善T2DM小鼠糖脂代谢紊乱,降低机体炎症,降低ET-1水平,显著下调主动脉ox-LDL、LOX-1、NF-κBp65、Caspase-3蛋白表达,显著上调eNOS蛋白表达;运动和联合干预增加NO、NO/ET-1水平,显著下调主动脉VCAM-1蛋白表达,二甲双胍对其未见显著性差异.3)跑台运动在降低ET-1,下调主动脉ox-LDL、LOX-1、VCAM-1蛋白表达方面较二甲双胍效果显著;联合干预在改善T2DM小鼠糖脂代谢,降低机体炎症,升高NO/ET-1,上调主动脉eNOS蛋白表达,下调ox-LDL、LOX-1、NF-κBp65、VCAM-1、Caspase-3蛋白表达较单一的跑台运动和(或)二甲双胍干预效果更为显著.结论:T2DM小鼠造模成功后未进行运动或药物干预,糖脂代谢紊乱进一步加重,血管舒张功能障碍,其可能机制主动脉炎性反应增强,血管细胞凋亡增加,导致血管损伤;8周跑台运动在降低血管炎症、改善血管舒张功能方面较二甲双胍作用显著,联合干预显示出更为有效的血管舒张功能改善效应.推测:8周跑台运动干预可能是通过下调ox-LDL、LOX-1蛋白表达,减轻血管炎症与细胞凋亡,从而改善血管舒张功能,联合干预则显示出运动与二甲双胍的协同作用.

目的:探讨有氧运动和二甲双胍对2型糖尿病大鼠血管炎症的影响及机制.方法:对2型糖尿病大鼠分别进行8周有氧运动,二甲双胍与二甲双胍+运动联合干预,检测干预后2型糖尿病大鼠糖脂代谢、主动脉形态、血管炎症标志物、氧化应激和NF-κB信号相关指标.结果:糖尿病对照组血清IL-1 β,TNF-α、MCP-l和VCAM-1浓度均显著高于正常对照组,有氧运动及联合干预能显著降低大鼠血清IL-1β、TNF-α、MCP-1和VCAM-1浓度;有氧运动及联合干预能够改善2型糖尿病大鼠血糖代谢,降低血管氧化应激,下调NF-κBp65蛋白和mRNA表达,上调IκBα蛋白表达.结论:有氧运动与联合干预可能部分通过改善2型糖尿病大鼠血糖代谢,降低血管氧化应激,抑制NF-κB信号缓解2型糖尿病大鼠血管炎症.

To investigate the effects of swimming exercise on vascular injury and oxidative stress state in prediabetic rat.Male Sprague Dawley rats were induced to the prediabetic model by high-energy emulsion,then they were randomly divided into four groups (sedentary control group,swimming control group,sedentary prediabetic group,swimming prediabetic group).Rats were sacrificed after 8 weeks' swimming training program.Malondialdehyde (MDA),superoxide-dismutase (SOD),peroxidase (POD),total antioxidant capacity (T-AOC),nitric oxide (NO) were detected in the serum and thoracic aorta.Vascular pathological changes were examined by HE staining.Compared with control group,in the serum and aorta of prediabetic rat,we found the visible vascular injury,significant oxidative stress state and remarkably decline of NO content.After eight weeks of swimming training,prediabetic rat showed obviously increase of the antioxidant capacity and NO content in serum and aorta and distinctly improving vascular damage.In conclusion,swimming training can balance oxidative stress state in prediabetic rat,so as to play a role of protection of blood vessels.

目的:评价有氧运动和二甲双胍对糖尿病大鼠血管炎症反应的影响.方法:对糖尿病大鼠(通过高脂饮食+腹腔注射链脲佐菌素制备)分别进行8周有氧运动、二甲双胍与二甲双胍和运动联合治疗干预,检测干预后糖尿病大鼠糖脂代谢、血管内皮和血管炎症标志物的相关指标,并探讨影响血管炎症反应的途径.结果:糖尿病对照组血清IL-1β、TNF-α和MCP-1浓度均明显高于正常对照组,二甲双胍组血清IL-1β、TNF-α和MCP-1浓度与糖尿病对照组相比没有明显变化,运动组大鼠血清IL-1β、TNF-α和MCP-1浓度显著低于糖尿病对照组,二甲双胍+运动组血清TNF-α浓度明显低于糖尿病对照组、二甲双胍组和运动组,MCP-1浓度明显低于糖尿病对照组.3个干预组血清IL-10浓度均高于糖尿病对照组,但仅有二甲双胍+运动组具有显著性差异.结论:有氧运动和二甲双胍+运动联合干预均能有效缓解糖尿病大鼠血管炎症反应.

目的:研究体育锻炼对大学生行走时步态参数的影响,为大学生参加体育锻炼提供理论依据.方法:利用h/p/cosmos gaitwayⅡ 测力跑台收集40名大学生步行时的步态参数.结果:长期参加体育锻炼的大学生步幅、步长、单脚支撑时长明显增加、双脚支撑时长、步态周期明显小于不参加体育锻炼的学生;长期参加体育锻炼的大学生学生最大力值、支撑力、蹬地率、明显高于不参加体育锻炼的学生,碰撞峰以及触地受力时长则减小.结论:长期参加体育锻炼的大学生步态在时空参数和运动学参数都具有显著性差异,长期参加体育锻炼有助于增加下肢肌力,改善步态稳定性.

In this study,we detected gene expressions of Frizzled 1-Frizzled 10 in the mice ovaries at different estrous stages based on real-time Q-PCR.Our results showed that they had different expression patterns except Frizzled 8.Expressions of Frizzled 1 and Frizzled 2 were the highest in proestrus;Frizzled 3 was higher in proestrus and diestrus than those in the other two periods;Frizzled 4 was found of high expression in all estrous cycle,which is highest in estrus;Other Frizzleds were all low at expression level.These results suggested that Frizzleds were widely and differentially expressed in estrous ovary and Frizzled 1-Frizzled 4 may play a more important role in the development of follicle during the estrous cycle.Our study provided a theoretical basis for revealing the role of Wnt signaling pathway in the process of ovary developement.

The purpose of this study was to optimize the procedure for cryopreservation of porcine embryos by vitrification.5-6 day-embryos(blastocyst/morula)were collected from superovulated Bama mini-pigs(sows/gilts).Different cryopreservation methods,cryopreservation tools,thining of zona pellucida(ZP) and recipient breeds were compared.The results were as follows:there were no significant different between two freezing method in embryo survival rate and cell number;the GMP vitrification method was more suitable and efficient in cryopreservation of pig embryos,the embryo survival rate(83.8% vs 77.6%) and blastocyst cell number(53.1 vs 47.5) were significantly higher than the OPS method(P<0.05);there were no significant differences in vitro survival rate by thinning the ZP(0.5% pronase,10 s) after warming but significantly improved the blastocyst cell number in surviving blastcysts(60.1% and 46.6%,P<0.01);the local pig breeds(Fengjing sows) are more suitable as recipients for vitrified/warmed blastcysts in the pregnancy rate and embryos efficiency(P<0.01).

The progesterone contents of peripheral serum of 5 Guizhou Xiang gilts and 5 Shanghai mini-gilts were determined during their oestrous cycles and early pregnancy by a radioimmunoassay method.It was found that the two gilt breeds during oestrous cycles were similar in the progesterone content change,and their peak values were 26.00 ngmL for Guizhou Xiang gilts and 24.36 ngmL for Shanghai mini-gilts and all appeared during 9~12 d of oestrous cycles.The change during early pregnancy was also largely alike.The two gilt breeds during-3~12 d of early pregnancy and-2~12 d of oestrous cycle were similar in the progesterone content change.On the 12th day of pregnancy,the peak value was 21.40 ngmL for Guizhou Xiang gilts and 27.78 ngmL for Shanghai mini-gilts.The progesterone content for both breeds slightly decreased after 12 days of pregnancy but increased again between 16~20 days of pregnancy and remained a relatively high level.The difference of the progesterone levels after16 days between the cycle and early pregnancy was significant and could be used to diagnose the gilt's early pregnancy.

The purpose was to optimize the vitrification for porcine embryos cryopreservation. Blastocyst/Morula (5-6th day-embryos) were collected from superovulated Bama mini-pigs (sows/gilts). We compared different cryopreservation methods, cryopreservation tools, thining of zona pellucida (ZP) and recipient breeds on the efficiency of porcine embryo cryopreservation. The results showed that: in embryo survival rate and blastocyst cell number, there were no significant differences between cryopreservation method I [embryos were vitrified by two step method with open pulled straw (OPS) and glass micropipette (GMP) in solution 1 (TCM199 + 20% FBS + 10% EG + 10% DMSO) for 3 min, and solution 2 (TCM199 + 20% FBS + 20% EG + 20% DMSO + 0.4 mol/L SUC) for 1 min, stored in liquid nitrogen] and method II[Blastocysts were cultured for 25 min in NCSU23 + 7.5 microg/mL cytochalasin B, centrifuged at approximately 13 000 xg for 12-13 min, and recovered back into pNCSU23. They were then equilibrated for 5 min in 2 mol/L ethylene glycol in pNCSU23, washed quickly in the vitrification medium, 8 mol/L ethylene glycol, 7% polyvinylpyrrolidone (PVP) in pNCSU23, loaded into OPS/GMP, and plunged into liquid nitrogen]. GMP vitrification method was more suitable and efficient than OPS method (P < 0.05) in embryo survival rate (83.8% vs 77.6%) and blastocyst cell number (53.1 vs 47.5) after thawing. Thining of ZP did not increase the survival rate, but significantly improved blastocyst cell number in the survival blastcysts (60.1 and 46, P < 0.01). Local pig breeds (Fengjing sows) were more suitable as recipients for embryo transfer of vitrified/warmed blastcysts, which can improve pregnant rate and embryo efficiency.

BMPs belong to the transforming growth factor-b superfamily. BMPs have been proved to have extensive biological functions in mammals, including growth regulation, cell proliferation and differentiation. More and more evidence has shown that BMPs play a key role in fertility, especially in folliculogenesis in female mammals. Smad proteins are intra-cellular signaling transduction molecules of BMP family, which can transduce the BMP signal from the cell membrane to the nucleus. In this review, BMPs, BMP/Smad-related signal transduction and the regulation of BMP activity were summarized, and the regulatory roles of BMP/Smad signal transduction pathway in folliculogenesis were discussed.

Porcine ovaries from a slaughterhouse were used as a test material,and the effects of different cryoprotectants,cryopreservation tools,in vitro maturation stages,treatments of CB Lipid removal and fertilization methods on the survival and development of vitrified porcine oocytes were investigated.The results showed that the ascending order of the toxicities of the osmotic cryoprotectants was EGGLDMSO.In the one-step vitrification of the MII-stage oocytes,the vitrification solution of EFS40 got the best result,and was significantly better than other vitrification solutions.For the time of treating the ooctyes with vitrification solutions,both 30 s and 1 min groups were not significantly different but significantly better than the 2 min group.Using GLT(55.8%) as cryopreservation tool got a significantly higher survival rate than using OPS(29.5%) and GMP(13.4%) methods.With the development of oocytes in the process of in vitro maturation,the anti-freezing ability of ooctyes increased.Compared with the control(53.6%),CB(55.6%) and Centrifugation+CB(54.5%) treatments could partly increase the survival rate of MII-stage oocytes after freezing,but lipid removal did not improve the survival rate of oocytes.Any cleavage from the vitrified MII-stage oocytes could not be got by in vitro fertilization method,but 6.1% cleavage rate could be got by the way of ICSI.

Jumonji, AT-rich interactive domain 1C (JARID1C) protein belongs to the highly conserved ARID protein family, which is involved in chromatin remodeling and transcriptional regulation during cell growth, differentiation, and development. In humans, this gene plays a vital role in normal brain development and function. Using an in silico approach in combination with 5' rapid amplification of cDNA ends (5' RACE), the full-length cDNA of JARID1C (GenBank accession No. EF139241) from porcine ovary, which contains 5,908 bp nucleotides, with an open reading frame (ORF) of 4,548 bp, has been cloned. The putative porcine JARID1C protein, which is located in the nucleus, encodes 1,516 amino acids with a molecular weight of 170 kDa and a pI of 5.44. Bioinformatic prediction indicates that the protein contains several conserved domains: a JmjN domain, an ARID domain, a JmjC domain, a C5HC2 zinc finger domain, and a PHD zinc finger domain. Similarity comparisons for nucleic and amino acid sequences reveal that the porcine JARID1C protein shares a high identity with its dog, mouse, rat, and human counterparts. The phylogenetic tree of the JARID1 subfamily proteins has been constructed to reveal the evolutionary relationship of various species. Real-time PCR analysis shows that the JARID1C gene is expressed in various tissues, but at different levels. The expression levels of this gene are higher in the brain and gonad than in other tissues, suggesting that the JARID1C protein plays a role in porcine brain and gonad functions.

In Bama-mini-pigs,five to six days post-insemination,morula and blastcysts were collected together after uterine.The purpose of this study was to vitrify zona pellucida-intact morula or blastcysts with OPS(open pulled straw) technology and obtain piglets after transfer in China.Blastocyst/morula were vitrified after a two-step equilibration in ethylene glycol,dimethyl sulfoxide and sucrose in Hepes-buffered TCM199+20%FBS.26 blastocyst/morula were loaded into OPS and plunged into liquid nitrogen.At embryo warming,a three-step dilution with decreasing concentrations of sucrose was applied.In each of 8 recipients,25-26 blastocyst/morula were transferes surgically.The pregnant recipient farrowed 8 alive piglets.Although survival rates are still compromised,OPS technology is therefore appropriate to cryopreserve porcine morula or blastcysts with intact zona pellucida.

1 卵母细胞与胚胎冷冻保存的意义 随着胚胎工程技术和低温生物学的不断进步,研究人员对哺乳动物的生殖细胞和组织的冷冻保存作了大量的研究.

291 embryos (Blastocyst/Morula) from 20 donor sows were vitrified by two step method with OPS (open pulled straw) in solution I (TCM199 + 20% FBS + 10% EG + 10% DMSO) for 3min, and solution II (TCM199 + 20% FBS + 20% EG + 20% DMSO + 0.4mol/L SUC) for 1min, stored in liquid nitrogen for 3 months, and transferred into 8 recipient sows after warming, one recipient sow was pregnant and 8 alive piglets were born. This is the first paper to report getting alive piglets by vitrification in China.

Trimethylsilyl pseudohalides Me3SiX, where X = NCS, NCO, or CN, were readily prepared conveniently in desirable yields by the reaction of Me3SiCl with NaX or KX catalyzed by PEG400 and zinc iodide under ultrasound irradiation.