表皮生长因子对葡聚糖硫酸钠诱导的结肠炎模型小鼠肠道损伤的修复研究

本文旨在讨论表皮生长因子(EGF)对葡聚糖硫酸钠(DSS)诱导的结肠炎模型小鼠肠道损伤的修复作用。选用24只6周龄BALB/c小鼠,随机分为3组,即:正常对照组、DSS模型对照组、DSS+EGF组。正常对照组小鼠饮用自来水;DSS模型对照组小鼠在试验第1~7天饮用5%DSS水溶液,第8~10天饮用自来水;DSS+EGF组小鼠按照DSS模型对照组处理,同时每天皮下注射EGF 2次,共注射10 d。结果表明:1)与正常对照组相比,DSS模型对照组小鼠结肠长度极显著降低(P0.01);与DSS模型对照组相比,DSS+EGF组小鼠结肠长度极显著增加(P0.01)。2)DSS模型对照组小鼠结肠可见典型溃疡,结肠损伤程度评分(CDS)极显著高于正常对照组(P0.01);DSS+EGF组小鼠结肠组织未见溃疡,与DSS模型对照组相比,CDS极显著降低(P0.01)。3)与正常对照组相比,DSS模型对照组小鼠结肠紧密连接蛋白(Occludin)浓度显著降低(P0.05);与DSS模型对照组相比,DSS+EGF组小鼠结肠Occludin浓度显著升高(P0.05)。4)与正常对照组相比,DSS模型对照组小鼠结肠白细胞介素-2(IL-2)和白细胞介素-4(IL-4)浓度极显著降低(P0.01),白细胞介素-10(IL-10)浓度显著降低(P0.05);与DSS模型对照组相比,DSS+EGF组结肠IL-2和IL-4浓度极显著增加(P0.01),IL-10浓度显著增加(P0.05)。由此可见,EGF可能通过提高肠道Occludin表达水平,调节肠道细胞因子浓度趋于正常水平,从而修复受损肠道组织,维持肠道黏膜屏障的完整性。     

In Vitro Comparison of a Novel External Fixator and Traditional Full‐Limb Transfixation Pin Cast in Horses

Objective: To compare the mechanical properties and failure modes of a standardized short oblique distal radial metaphyseal osteotomy stabilized using either a transfixation pin cast (TPC), a modular‐sidebar external skeletal fixator (ESF), or a solid‐sidebar ESF (modular‐ or solid‐ESF, respectively) using static or cyclic axial loading to failure. Study Design: In vitro study. Animals: Equine cadaver forelimbs. Methods: A 30° oblique distal radial osteotomy was created and stabilized using 1 of the 3 fixation methods: (1) TPC, (2) modular‐ESF, or (3) solid‐ESF. Limbs were tested using static (TPC, modular‐ESF, and solid‐ESF) or cyclic (TPC and solid‐ESF) axial loading to failure. The stiffness, yield load, yield displacement, failure load, and failure displacement for static loading and the cycles to failure for cyclic loading at 75% failure load were obtained. Data were analyzed using a Kruskal–Wallis test. Level of significance was P<.05. Results: The solid‐ESF had a greater stiffness, higher yield and failure load and a lower yield and failure displacement than the TPC (P=.01) and the modular‐ESF (P=.02). TPC had a higher yield load, failure load, and yield displacement than the modular‐ESF (P=.01). Mean cycles to failure for TPC was 2996±657 at a load of 16,000 N and for solid‐ESF 6560±90 cycles at a load of 25,000 N. Conclusions: The solid‐ESF was stiffer and stronger than the TPC and modular‐ESF and failed at a greater number of cycles in axial loading compared with the TPC. Clinical Relevance: This study is an initial step in evaluating the solid‐ESF. Further testing needs to be performed, but this fixation may offer a viable alternative to the traditional TPC for stabilization of long bone fractures in adult horses.     

In vitro biomechanical properties of linear, circular, and hybrid external skeletal fixation devices for use in large ruminants

OBJECTIVE: To report the biomechanical properties of 3 external skeletal fixation (ESF) devices for use in large ruminants. STUDY DESIGN: In vitro biomechanical testing of ESF constructs. SAMPLE POPULATION: Adult buffalo (weighing, 250-350 kg) tibiae (n=27). METHODS: ESF constructs (bilateral linear fixator [BLF], 4-ring circular external fixator [CEF], and hybrid fixator [HF]) were made using mild (low carbon) steel implants plated with nickel and cadaveric buffalo tibiae. After ESF application, a 1 cm mid-diaphyseal gap was created. Constructs were loaded to failure, on a materials testing machine, in axial compression (n=5/ESF type) and craniocaudal bending (n=3/ESF type). In addition, 3 CEF constructs were tested in intact tibiae under craniocaudal bending. RESULT: In compression, HF was the strongest and most rigid construct; yield load was significantly higher for HF than for BLF or CEF. Under bending, both CEF and HF had similar strength and modulus of elasticity. Strength for BLF was higher than CEF and HF, whereas the reverse was true for modulus of elasticity. CONCLUSIONS: ESF made from mild steel for use in large ruminants could withstand相似文献   

Protein Synthesis during Maturation of Bovine Oocytes, Effect of Epidermal Growth Factor

The objective of this study was to determine whether epidermal growth factor (EGF) affected protein synthesis in oocytes during maturation. Initially, the effect of EGF on oocyte maturation was examined to ensure that there was a beneficial effect of EGF in the protein-free maturation medium used in these studies. Results showed that the presence of EGF during maturation significantly enhanced cleavage rate and development to the blastocyst stage. Development after maturation in the presence of EGF was similar to that seen in medium containing serum, luteinizing hormone, follicle-stimulating hormone and estradiol. Protein synthesis was examined in immature oocytes and after 16 or 24 h maturation. Oocytes from each group were labelled by incubation for 4 h with ³⁵S-methionine, the proteins were then separated by two-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Between 400 and 500 proteins could be separated using this method and marked changes in protein synthesis was observed during maturation. Changes in eight different proteins were observed when protein patterns from oocytes matured for 16 h with and without EGF were compared. These results suggest that EGF plays a physiological role in oocyte maturation and identification of the proteins induced by EGF could be important for improving our understanding of oocyte maturation in vitro .     

Effects of glucagon‐like peptides 1 and 2 and epidermal growth factor on the epithelial barrier of the rumen of adult sheep

Epidermal growth factor (EGF) and glucagon‐like peptides (GLP) modulate the tight junctions (TJ) of the intestinal epithelial barrier (EB) of monogastric animals. This work tried to elucidate whether GLP‐1, GLP‐2 and EGF can affect the EB of the rumen. Ovine ruminal epithelia were incubated in Ussing chambers for 7 hr with 25 or 250 nM of either GLP‐1 or GLP‐2 on the serosal side, with 2.5 nM of EGF on the serosal side or with 0.25 or 2.5 nM EGF on the mucosal side. No treatment affected tissue conductance. Short‐circuit current (I_sc) was affected by time and treatment and their interactions. Only 250 nM of either GLP‐1 or GLP‐2 decreased I_sc in certain periods compared with 25 nM GLP‐1 or 0.25 nM mucosally applied EGF; however, not when compared to control epithelia. Fluorescein flux rates (J_fluor) of ruminal epithelia were affected by treatment, time and time × treatment interaction. The time × treatment interaction was based on an increase in J_fluor between the first and last hour in epithelia incubated with 25 nM GLP‐1 or GLP‐2 and in epithelia incubated with EGF. After 7 hr incubation, claudin‐7 mRNA expression was downregulated in all treatments. Claudin‐1 mRNA was upregulated after incubation with 2.5 nM EGF on the serosal side, claudin‐4 mRNA was downregulated by 2.5 nM EGF on the mucosal side, and occludin mRNA was increased after incubation with 250 nM GLP‐2. The protein abundance of all tested TJ proteins was not influenced by treatment. We conclude that GLP‐1, GLP‐2, and EGF have no obvious acute effects on the EB of ruminal epithelia under simulated physiological conditions ex vivo. However, by decreasing the mRNA expression of claudin‐7 and partly affecting other TJ proteins, they may modulate EB in the longer term or under certain conditions.     

Biomechanical evaluation of acrylic external skeletal fixation in dogs and cats.

Acrylic external skeletal fixators (ESF) were compared with Kirschner ESF in biomechanical tests. A 2-cm unilateral acrylic ESF was found to be superior to medium Kirschner ESF in compression and shear loads. Acrylic ESF performed as well as Kirschner ESF in torsion loads. Acrylic ESF were used on 11 dogs and cats for repair of long bone fractures, for arthrodesis, or for immobilization of joints following ligament or tendon surgery. There were no complications associated with the use of acrylic ESF. Acrylic ESF offers the advantage of reduced cost, improved versatility, and simplified application technique when compared with Kirschner ESF.     

Biomechanical analysis of 3 fixation techniques in rabbit radius and humerus bones

The objective of this study was to compare the strength and stiffness of various fixation methods applied to the long bones of the rabbit forelimb. Twenty rabbit radius/ulna and 20 rabbit humeri were randomly assigned to 1 of 4 groups. Control bones remained intact, whereas all others were osteotomized to create fracture models that were fixated with locking plate and locking screws (LP), veterinary cuttable plate (VCP) with cortical screws, or external skeletal fixator constructs (ESF), and tested in 4-point bending until failure. Load/deformation curves were generated for each sample and used to calculate stiffness (slope of the curve) and strength (load to failure). Intact controls had greater strength and stiffness than any fixation techniques in the rabbit radius/ulna and humeri samples. Locking plate and VCP constructs had greater stiffness than ESF when applied to the radius, whereas locking plate constructs were stronger than VCP or ESF when applied to the humerus. Overall, the LP construct had characteristics most closely resembling those of the intact control regarding strength in the humerus. Therefore, fracture fixation with a LP would provide the greatest strength in humeral fracture repairs in the rabbit.     

鸡柔嫩艾美耳球虫不同抗药性虫株的种内多态性研究

应用RAPD技术进行柔嫩艾美耳球虫4个单一抗药性虫株与1个敏感株的基因组DNA多态性分析，发现4个抗药性虫株及敏感株之间的相似值均大于99%；OPAO3引物对抗盐霉素株扩增出一条约500bp的特异条带，OPH02引物对抗克球粉株和抗盐霉素株均扩增出了约1kb的第三条主带，这些特异条带的出现很可能与球虫抗药性基因有关，有可能用于抗药性虫株的诊断与鉴定。     

Unilateral Uniplanar External Skeletal Fixation for Isolated Diaphyseal Tibial Fractures in Skeletally Immature Dogs

Objective: To report and evaluate use of external skeletal fixation (ESF) for repair of isolated (fibula intact) diaphyseal tibia fractures in skeletally immature dogs.
Study Design: Case series.
Animals: Skeletally immature (mean age, 17 weeks; range, 12–23 weeks) dogs (n=5) with isolated diaphyseal tibial fractures.
Methods: Medical records (2006–2007) of 5 dogs with isolated diaphyseal tibial fractures treated with Type 1a ESF were reviewed and outcome assessed by clinical examination and telephone interviews.
Results: Dogs were evaluated ∼2 and 4 weeks after surgery. Limb function, muscle mass, and comfort level remained very good to excellent throughout healing. Bony union was confirmed radiographically and the ESF removed ∼4 weeks after surgery (mean, 31.4 days; range, 28–37 days). Owner satisfaction was high when contacted a minimum of 4 weeks after ESF removal.
Conclusions: Use of Type Ia ESF with positive profile threaded pins is a practical and biologic method for treatment of isolated tibial fractures in skeletally immature dogs
Clinical Relevance: ESF is an effective alternative treatment to external coaptation and internal fixation for isolated diaphyseal tibial fractures in skeletally immature dogs.     

Evaluation of a nontoxic rigid polymer as connecting bar in external skeletal fixators

OBJECTIVE: To investigate the mechanical characteristics of a nontoxic, low-cost, rigid polymer (RP) and to compare the structural and mechanical properties of a full-frame external skeletal fixator (ESF) with either RP connecting bars, polymethylmethacrylate (PMMA) connecting bars, or stainless-steel (SS) clamps and connecting bars. STUDY DESIGN: In vitro mechanical evaluation. METHODS: Mechanical properties were assessed using an in vitro bone fracture model with a bilateral uniplanar ESF (type II). Identical ESF were built with connecting bars using RP (n = 8), PMMA (n = 8), and SS connecting bars and clamps (System Meynard; n = 3). Nondestructive mechanical tests were performed in uniaxial compression (AC) and craniocaudal (CC) 4-point bending, as well as fatigue AC. Composite stiffness for each specimen and for each loading mode was calculated from 6 replicate measures using the slope of the load displacement curve at small displacements. RESULTS: RP, PMMA, and SS ESF constructs yielded mean +/- SD composite stiffness values of 227 +/- 15, 381 +/- 30, and 394 +/- 9 N/mm in AC and of 35 +/- 2, 24 +/- 2, and 15 +/- 0 N/mm in CC, respectively. CONCLUSIONS: Structural and mechanical properties of RP are satisfactorily rigid and fatigue resistant for its use as a connecting bar in ESF. CLINICAL RELEVANCE: RP connecting bars in an ESF are a reliable, versatile, nontoxic and inexpensive option for the veterinary surgeon.     

Middle upper beak fracture in a Red-crowned crane that completely recovered with external skeletal fixation

An adult Red-crowned crane in captivity that had a displaced compound fracture of the middle upper beak caused by an accident was treated by using Type I-a external skeletal fixator (ESF). The ESF that was equipped with a unilateral epoxy putty fixator and with five half-pins was fixed on the premaxilla bone. The crane with the ESF on the beak was able to feed smoothly by itself. The ESF was removed 49 days after the first surgery. Beak malocclusion, which was observed in the latter half period of fixing, spontaneously improved within one month after removal of the ESF. This is the first successful case of repair of an upper beak fracture in a Red-crowned crane.     

小鼠精原干细胞体外培养体系建立及功能鉴定

旨在通过探索ICR品系小鼠精原干细胞(spermatogonial stem cells,SSCs)的体外稳定培养,为别的品系小鼠和大动物的SSCs体外培养提供参考.本研究采集6?8日龄ICR乳鼠的睾丸,用两步酶消化法(胶原酶和胰蛋白酶)和差速贴壁法来纯化SSC细胞.用不同的饲养层(小鼠胎儿成纤维细胞(mouse em...     

表皮生长因子对成年水牛晶状体上皮细胞促增殖作用

为观察表皮生长因子 ( epidermalgrowth factor,EGF)对成年水牛晶状体上皮细胞增殖作用的影响。将不同浓度 EGF作用于体外培养的成年水牛晶状体上皮细胞 ,采用 MTT法测定细胞的增殖能力。结果 :EGF在浓度为 1ng/ m L和 1 0 ng/ m L作用的前 3 d促增殖作用逐渐加强 ,呈现时间依赖性 ,且在第 3 d达到最大促增殖效果。不同浓度的 EGF对晶状体上皮细胞的增殖作用不同 ,浓度在 1 0 ng/ m L作用 2 4h后即有明显的促增殖作用 ( P <0 .0 5) ,而作用72 h后最低的有效浓度为 1 ng/ m L,而且 2 50ng/ m L EGF有最大促增殖作用。结论 :EGF是诱导成年水牛晶状体上皮细胞增殖的重要因素。     

External skeletal fixation in the management of equine mandibular fractures: 16 cases (1988-1998)

Fifty-three cases of equine mandibular fractures were managed surgically from 1988-1998, of which 16 (30%) were repaired by external skeletal fixation (ESF). Three surgical methods were utilised: transmandibular 4.76 or 6.35 mm Steinmann pins incorporated into fibreglass casting material or nonsterile dental acrylic (methyl methacrylate - MMA) bars reinforced with steel; transmandibular 9.6 mm self-tapping threaded pins +/- 4.76 or 6.35 mm Steinmann pins incorporated into MMA bars reinforced with steel; and 4.5 mm or 5.5 mm ASIF cortical bone screws incorporated into MMA bars reinforced with steel or a ventral MMA splint. Fourteen horses were presented to the hospital for fixator removal at an average of 56.2 days. At removal, fractures were stable and occlusion of incisor and cheek teeth was considered adequate. Complications of the procedure occurred in 3 horses. Two horses with persistent drainage and ring sequestra from pin tracts required curettage 4 or 5 months after ESF removal. A third horse required replacement of the original fibreglass ESF with MMA bars to regain access to open, infected wounds. Another horse required removal of the second premolar at the time of fixator removal because the tooth root had been damaged in the original injury. ESF for the surgical management of mandibular fractures in horses has produced good results, with incisive and cheek tooth alignment reestablished in all horses. Horses that were managed via ESF had a rapid return to full feed and did not require any supplementation via nasogastric tube or oesophagostomy to maintain bodyweight or hydration status.     

饲用玉米根际促生菌资源筛选及其特性研究

为获得玉米根际促生菌(PGPR)菌株并明确其功能特性;以玉米根系及根际土壤为材料,利用选择性培养基分离筛选溶磷菌与固氮菌,测定所选菌株溶磷能力和固氮酶活性,并测定其产IAA的能力,从中筛选综合性能优良的菌株,在此基础上,运用生理生化鉴定和16SrDNA分子生物学鉴定相结合的方法鉴定优良菌株的种属。结果表明:玉米根际有大量PGPR菌,分布特征表现出根表(RP)根表土(RS)远根土(NRS)根内(HP)的数量分布规律,呈根际效应。最终获得262株PGPR菌,其中固氮菌48株,溶解无机磷细菌108株,溶解有机磷细菌106株,有分泌IAA能力的菌株15株。筛选出综合性能优良、有进一步开发应用潜力的菌株7株。经鉴定其中3株为Pseudomonas fluorescens;2株为Bacillus subtilis;Klebsiella oxytoca和Bacillus megaterium各1株。     

The development of a transport and enrichment medium for Campylobacter fetus

A transport and enrichment medium was developed for Campylobacter fetus. From inocula of between 10 and 35 organisms the medium was able to support the multiplication of 19 of 21 strains of C. fetus if the medium was incubated immediately after inoculation; when incubation was delayed for 3 days after inoculation, only seven of the 21 strains multiplied. From inocula of 100-350 organisms all 21 strains multiplied following immediate incubation, and 20 of 21 when incubation was delayed for 3 days. From inocula of about 10(4) organisms all strains multiplied following immediate or delayed incubation. The medium restricted the growth of Proteus vulgaris and Pseudomonas aeruginosa.     

Effects of epidermal growth factor on atrophic enteritis in piglets induced by experimental porcine epidemic diarrhoea virus

Epidermal growth factor (EGF) promotes gastrointestinal mucosal recovery by stimulating the mitogenic activity of intestinal crypt epithelial cells. The aim of this study was to determine the effects of EGF on atrophic enteritis induced in piglets by experimental infection with porcine epidemic diarrhoea virus (PEDV) strain Dr13. Two groups of 12 conventional, colostrum-deprived, 1-day-old, large White-Duroc cross breed piglets were inoculated orally with PEDV (3 x 10(5) 50% tissue culture infective doses), with or without EGF (10 microg/kg/day, intraperitoneally once daily for 4 days after infection) and compared to 12 uninfected, untreated control piglets. PEDV+EGF piglets had less severe clinical signs than PEDV only piglets at 48 and 60 h post-infection (hpi). Histologically, the ratio of villous height:crypt depth of PEDV+EGF piglets was significantly higher than PEDV only piglets at 36 and 48 hpi. Immunohistochemistry for Ki67 demonstrated increased proliferation in intestinal crypt epithelial cells of PEDV+EGF piglets compared to PEDV only piglets at 36, 48 and 60 hpi. EGF stimulates proliferation of intestinal crypt epithelial cells and promotes recovery from atrophic enteritis in PEDV-infected piglets.     

Low Incidence of an Altered Endometrial Epidermal Growth Factor (EGF) Profile in Repeat Breeder Holstein Heifers and Differential Effect of Parity on the EGF Profile Between Fertile Holstein (Dairy) and Japanese Black (Beef) Cattle

A high incidence (about 70%) of alteration in endometrial epidermal growth factor (EGF) profile, i.e., loss of 2 peaks on days 2–4 and 13–14, has been linked to a reduced fertility in multiparous repeat breeder Holstein cows. However, the EGF profile in Holstein heifers and other breeds (types) of cattle has not been investigated. In study 1, EGF concentrations were determined using endometrial tissues obtained by biopsy on days 3, 7 and 14 from 84 fertile Holstein heifers to obtain a normal range and 53 repeat breeder Holstein heifers to estimate incidence of alterations in the EGF profile. In repeat breeder heifers, EGF concentrations were similar to fertile controls on 3 days and five animals (9.4%) had an altered EGF profile with EGF concentrations below the normal range on days 3 and 14. In study 2, EGF concentrations on day 3 were repeatedly examined from the nulliparous period to the third postpartum period in 28 Holstein (dairy) and 47 Japanese Black (beef) cattle. The effect of parity on EGF concentrations on day 3 was different between Holstein and Japanese Black cattle. In Japanese Black cows, the EGF concentrations were consistently high throughout the study period, while in Holstein cows, the EGF concentrations decreased after the second calving. In conclusion, unlike multiparous repeat breeder Holstein cows, an altered EGF profile may not be a major cause of repeat breeding in Holstein heifers, and the peak EGF concentrations around day 3 may decrease even in fertile populations of multiparous dairy cows, but not in beef cows.     

A progestin-based treatment with a high dose of estradiol benzoate normalizes cyclic changes in endometrial EGF concentrations and restores fertility in repeat breeder cows

The objective of the present study was to examine the efficacy of a progestin-based treatment with a high dose of estradiol benzoate (EB) to normalize the epidermal growth factor (EGF) profile in the uterine endometrium and restore fertility in repeat breeder cows. Repeat breeder cows without peaks in their endometrial EGF concentrations on Days 3 and 14 of the estrous cycle were used throughout the study. The effect of 1 (standard dose), 2.5 and 5 mg of EB in a progestin-based treatment protocol (EB1, EB2.5 and EB5 treatments, respectively; n=5 for each group) on endometrial EGF concentrations was first examined. The EB1 and EB2.5 treatments in the repeat breeder cows produced a suppressed response in endometrial EGF compared with EB1 treatment in the fertile controls (n=5) and failed to restore the normal EGF profile during the next estrous cycle. However, EB5 treatment produced an increase in EGF concentrations similar to the fertile controls and normalized the endometrial EGF profile. The effects of the EB1 and EB5 treatments (n=30 for each group) on the endometrial EGF profile and fertility were then examined in the repeat breeder cows. The proportion of cows, with an EGF profile normalized by the treatments was higher in the EB5 group (66.7%) than in the EB1 (30.0%) and untreated control (13.3%; n=30) groups (P<0.01). The pregnancy rates of the cows having a normal EGF profile after treatment in the EB1 and EB5 groups were similar (88.9 and 85.0%, respectively) and higher than those of the cows having an abnormal profile within the same groups (19.0 and 30.0%, respectively, P<0.01). In summary, the endometrial response to EB in terms of the EGF concentration was suppressed in repeat breeder cows. A high dose (5 mg) of EB in a progestin-based treatment was found to be effective for restoration of a normal EGF profile and fertility in repeat breeder cows having lesser endometrial EGF concentrations on Days 3 and 14.     

Strain differences in whole-body protein turnover in the chicken embryo.

1. Whether or not there is a strain difference in embryonic whole-body protein turnover rates was tested using the chicken embryos of Rhode Island Red carrying a sex-linked dwarf gene (dwarf), White Leghorn (layer), and White Cornish X White Plymouth Rock (broiler) strains on day 12 of incubation. 2. Whole-body protein synthesis was estimated by injecting L-[15N]-phenylalanine either intraperitoneally or intravenously on day 12 of incubation in order to investigate the effect of the route of isotope administration. The results showed that the values for fractional and absolute synthesis rates were approximately 13% higher by intravenous injection than by intraperitoneal injection. 3. Whole-body protein turnover, both in terms of fractional and absolute rates, was significantly faster in dwarf than in broiler embryos, with intermediate values in layer embryos, although no growth differences were observed on day 12. 4. Difference in egg weight, measured before incubation, did not affect protein turnover. 5. It was concluded that the strain difference manifested in whole-body protein turnover of the chicken embryo would probably be a reflection of differences in genetic background.