Genotype evaluation,muscle histopathology and ultrastructure in a Duchenne muscular dystrophy animal model

AIM: To evaluate the genotype , muscle histopathology and ultrastructure in dko mice. METHODS: Dystrophin/Utrophin-deficient double knockouts (dko) mice were obtained from university of Oxford, UK. Genotype of filial generation of heterozygote was evaluated by PCR-SSP. HE staining and fluorescent immunohistochemistry by SABC-Cy3 were used to detect striated muscle of dko mouse, and the muscle ultrastructure was observed by transmission electron microscope(TEM). RESULTS: In 112 filial generation mice, there were 28 mdx (25.0%), 26 dko (23.2%) and 58 heterozygote (51.8%), which coincided with the law of Mendelian inheritance. HE staining showed that the myocytes were not very uniform, there were phenomenon of round outline, centrally nucleated fibers, widening interspace, inflammatory cell infiltration and connective tissue proliferation in dko mice. There were no any immunofluorescent expression of dystrophin and utrophin in sarcolemma in dko mice. TEM showed sarcolemma breakage, separation and edema, and loose myofibril texture, inflammatory cell infiltration and connective tissue proliferation in dko mice. CONCLUSION: PCR-SSP is a very quick and accurate way for genotype evaluation of filial generation. The pathophysiology of dko mouse was very similar to Duchenne muscular dystrophy (DMD), and dko mouse is an ideal animal model for study of DMD clinical therapy.     

Influence of irbesarten on renal hypertrophy in streptozotocin-induced diabetic rats LIU Bi-cheng,LUO Dong-dong,ZHANG Xiao-liang.

AIM: To investigate the influence of irbesartan (Irb), a new angiotensin II receptor 1 antagonist, on renal hypertrophy in streptozotocin (STZ)-induced diabetic rats. METHODS: Sprague-Dawley (SD) rats were randomly divided into three groups: normal group (Group N, n=7), diabetic group (Group DN, n=6) and irbesartan treated group (Group DNI, n=7). In the experimental group, after the rats subjected to uninephrectomy, STZ was given by peritoneally injection at bolus dose of 50 mg/kg to induce diabetes. Blood glucose (BG), body weight (BW), urinary albumin excretion (Ualb), 24 hour proteinuria (24 h Upro) were measured at week 4, 8, 12, respectively. By the end of experiment at week 12, creatinine clearance (Ccr), kidney weight (KW), indicator of renal hypertrophy (KW/BW), renal total protein content (RTP), glomerular area (AG) and glomerular volume (VG) as well as glomerular basement membrane (GBM) were determined by semi-quantitative pathology technique. RESULTS: It was showed that there was no significant difference in BG between group DN and DNI, while Irb significantly reduced the increasing of Ualb, 24 h Upro in diabetic rats compared to control group (P＜0.01, respectively). Furthermore, Irb markedly inhibited the increasing of KW, KW/BW, RTP, A_G, V_G in diabetic rats (P＜0.05, P＜0.01, respectively). Of interest, Irb significantly prevented the increasing of GBM in diabetic rats. CONCLUSION: Irb exerts its early renal protective action by reducing proteinuria and inhibiting renal hypertrophy as well as the thickening of GBM.     

A novel mutation of the CLCN1 gene associated with myotonia hereditaria in an Australian cattle dog

BACKGROUND: Heritable myotonia is a genetic muscle disorder characterized by slow relaxation of skeletal muscles. The main clinical signs are skeletal muscle stiffness, especially after vigorous contraction, and muscle hypertrophy. Muscle stiffness may be enhanced by inactivity, and often is relieved by exercise. Myotonia can be inherited in an autosomal dominant or recessive manner (Thomsen- or Becker-type myotonia, respectively). In mice, goats, Miniature Schnauzer dogs, and most affected humans, the disorder is caused by mutations in CLCN1, which encodes the skeletal muscle voltage-gated chloride channel, Cl1C-1. HYPOTHESIS: We hypothesized that an Australian Cattle Dog with generalized muscle stiffness and hypertrophy examined at the Ontario Veterinary College would have a mutation in the CLCN1 gene. ANIMALS: A pure-bred Australian Cattle Dog from Ontario, Canada, was used. METHODS: Based on clinical signs and electromyographic test results, a diagnosis of myotonia hereditaria was made, and a muscle biopsy was collected for genetic analysis. RESULTS: Sequence data obtained from the affected dog confirmed that it was homozygous for a single base insertion in the CLCN1 coding sequence. This mutation would result in a truncated ClC-1 protein being expressed, which, based on molecular evidence from other studies, would result in functionally compromised chloride conduction in the skeletal muscles of the animal. CONCLUSIONS AND CLINICAL IMPORTANCE: To the authors' knowledge, this report describes the Ist case of myotonia in an Australian Cattle Dog and represents the 1st non-Schnauzer canine myotonia to be genetically characterized. In addition, we developed a polymerase chain reaction-based genetic screen to detect heterozygotes with this mutation in the at-large Australian Cattle Dog population.     

不同饲养方式对肉鸡肌纤维组织学特性及肌肉嫩度影响的研究

[目的]研究了不同饲养方式对河田鸡肌纤维组织学特性及肌肉嫩度的影响,为肉鸡品质调控提供理论依据。[方法]研究户外散养、室内平养、室内笼养3种饲养方式对不同日龄河田鸡腿肌和胸肌肌纤维组织学特性的影响及其与肌肉嫩度的关系。[结果]结果表明,户外散养鸡的胸肌和腿肌肌纤维直径和肌肉剪切力显著大于笼养鸡（P〈0．05）,而肌纤维密度显著低于笼养鸡（P〈0．05）。随着日龄的增加,肌纤维的直径和肌肉剪切力显著增加,而肌纤维密度显著下降。公鸡的肌纤维直径和剪切力大于母鸡。相关性分析表明,胸肌肌肉剪切力与肌纤维直径呈显著正相关（P〈0．05）,而与肌纤维密度呈负相关（P〈0．05）;腿肌肌肉剪切力与肌纤维直径和密度关系不显著（P〉0．05）。[结论]饲养方式对肉鸡肌纤维的组织学特性有显著影响,户外散养促进肉鸡肌纤维发育,相应增加了肌肉剪切力。     

肌肉苹果酸酶活性与猪肉品质关系的研究

[目的]探讨肌肉苹果酸酶活性与猪肌肉品质的关系,为提高猪肌肉品质提供理论依据。[方法]选取3个不同品种的猪（鄂西黑猪、长野鄂杂交猪和长大二元杂交猪）,比较了肌肉苹果酸酶活性以及肉质性状之间的差异。[结果]结果表明,鄂西黑猪的肌肉苹果酸酶活性和肌内脂肪显著高于长野鄂（三元）杂交猪和长大二元杂交猪（P〈0.05）;长大二元杂交猪的肌肉pH2值、滴水损失和剪切力显著高于鄂西黑猪（P〈0.05）。[结论]猪肌肉苹果酸酶活性对猪肉品质存在一定的影响。其中,肌肉苹果酸酶活性与肌内脂肪含量呈显著正相关,与肉色呈正相关,而与滴水损失和剪切力呈显著负相关。     

The effects of 17β-estradiol on cardiomyocyte hypertrophy induced by endothelin

AIM:To investigate the effect of 17β-estradiol (E₂) on myocardial hypertrophy induced by endothelin-1(ET-1) and the related mechanism. METHODS:Myocardial cells from neonate rats were cultured in vitro and myocardial hypertrophy model was established with ET-1.The effects of 17β-estradiol on myocardial hypertrophy were observed. The role of ERK1/2 in the effects of 17β-estradiol was also detected. RESULTS:Compared with control group,ET-1 increased cell protein content,cell surface area and ［³H］^-Leucine ［³H］^-Leu) incorporation. Pretreatment with E₂ for 24 h could inhibit the increase in cell protein content,cell surface area and ［³H］^-Leu incorporation induced by ET-1.ET-1 significantly stimulated ERK1/2 activity,which was prevented by pretreatment with E₂.Tamoxifen,estradiol receptor antagonist,partially inhibited the effect of E₂. The ability of ET-1 to stimulate ［³H］^-Leu incorporation was significantly blocked by PD98059,which could enhance the inhibitory effect of E₂ on the increase of ［³H］^-Leu incorporation in cardiomyocytes induced by ET-1.CONCLUSION:E₂ can inhibit cardiomyocyte hypertrophy induced by ET-1. This effect is mediated by estrogen receptor. ERK1/2 signal pathway is closely correlated with the inhibitory effect of E₂ on cardiomyocyte hypertrophy induced by ET-1.     

Effects of atorvastatin on cardiac myocyte hypertrophy of neonatal rat induced by angiotensinⅡ in vitro and TLR4 expression

AIM: To assess effects of atorvastatin (Ator) on cardiac myocytes (CM) hypertrophy of neonatal rat induced by angiotensinⅡ (AngⅡ) in vitro and Toll like receptor 4 (TLR4) expression for theoretical bases of preventing and treating myocardial hypertrophy.METHODS: CM of neonatal Sprague-Dawley (SD) rats were isolated with trypsin digestion method and those growth-arrested cells were stimulated with 10^-7 mol/L AngⅡ in the presence of various concentrations of Ator.The method of coomassie brilliant blue was adopted to evaluate the protein contents of CM.The changes in β-MHC,AT₁ receptor and TLR4 mRNA expression were observed by RT-PCR.RESULTS: ① AngⅡ increased the protein content of CM and β-MHC mRNA expression significantly,upregulated AT₁ receptor and TLR4 gene expression.② In a dose-dependent manner,Ator inhibited the increases in the protein contents of CM and β-MHC mRNA expression induced by AngⅡ.③ In a dose-dependent manner,Ator downregulated the AT₁ receptor and TLR4 mRNA expression of CM hypertrophy of neonatal rat induced by AngⅡ.CONCLUSION: Ator inhibits CM hypertrophy,downregulates the AT₁ receptor and TLR4 gene expression.     

双氢睾酮诱导大鼠心肌细胞肥大模型的建立

应用胰蛋白酶分次消化法分离乳鼠心肌细胞,以差速贴壁法纯化心肌细胞,α-sarconme-actin抗体免疫细胞化学染色鉴定心肌细胞。心肌细胞在无血清无酚红培养基中培养48h后,用双氢睾酮(DHT)诱导心肌细胞肥大,建立心肌细胞肥大模型。24h后检测心肌细胞肥大的指标心肌细胞表面积;BCA法测定心肌细胞蛋白含量;半定量RTPCR两步法检测心肌细胞肥大的特征性基因—心房利钠因子(atrial natriuretic factor,ANF,β-肌球蛋白重链(β-myosin heavy chain,β-MHC)mRNA的表达。结果显示免疫细胞化学染色显示培养的心肌细胞纯度达到90%以上,心肌细胞分离良好。与对照组相比,10-8 mol/L的DHT能显著的增加心肌细胞表面积、蛋白质含量、ANP和β-MHC基因表达的增加(P0.01),心肌细胞肥大模型建立成功。     

Effects of unilateral ovariectomy on recruitment and growth of follicles in the rainbow trout,Oncorhynchus mykiss

Virgin female rainbow trout, Oncorhynchus mykiss, were unilaterally ovariectomised at various stages of ovarian development to investigate the effect of the removal of one ovary on subsequent recruitment and growth of follicles in the remaining ovary. The right ovary was removed from groups of 12–15 fish, 12, 7 and 4 months before they were due to ovulate, and the gonadosomatic index and follicle number and size determined just prior to ovulation. There were no differences in fecundity or follicle size in fish unilaterally ovariectomised at 12 and 7 months prior to ovulation compared to the controls. However, in the females unilaterally ovariectomised 4 months prior to ovulation, the remaining ovary either had the normal number of follicles for a single ovary but of a significantly larger size than follicles in the controls, or alternatively had almost 70% more than the normal number of vitellogenic follicles but comprising two distinctly different size populations. Differences in plasma oestradiol-17β concentrations at the final sample were seen only in females unilaterally ovariectomised 4 months prior to ovulation, where the levels were significantly lower than both the sham operated and control fish (p < 0.05). These data show that in the rainbow trout, complete compensatory ovarian hypertrophy following unilateral ovariectomy can occur throughout a major part of ovarian development, but that follicle recruitment is limited to stages up to (and therefore fecundity is determined by) mid-vitellogenesis.     

长期服用氨氯地平对高血压病降压的疗效及左室结构功能的影响

目的；观察氨氯地平治疗高血压病的长期降压效应及左室结构功能的改变。方法：２６例患者日服氨氯地平５－１０ｍｇ，平均２４个月。于服药前，服药后４周及治疗结束时进行偶测血压，动态血压及彩色多普勒超声心动图检查。结果：治疗后偶测血压，白昼及夜间动态血压明显平稳下降，心率则差异无统计学意义，室间隔，左室后壁厚度及左室重量指数均显著下降。