Co-treatment with surfactant and sonication signifi cantly improves Agrobacterium-mediated resistant bud formation and transient expression efficiency in soybean

Soybean is a widely planted genetically modified crop around the world. However, it is still one of the most recalcitrant crops for genetic transformation due to the difficulty of regeneration via organogenesis and some factors that affect the transformation efficiency. The percentages of resistant bud formation and transient expression efficiency are important indexes reflecting the regeneration and transformation efficiency of soybean. In this study, the percentages of resistant bud formation and transient expression of 13-glucuronidase （GUS） were compared after treatment with sonication or surfactant and co-treatment with both. The results showed that treatment with either sonication or surfactant increased the percentage of resistant bud formation and transient expression efficiency. The highest percentages were acquired and significantly improved when cotyledon node explants were co-treated with sonication for 2 s and surfactant at 0.02% （v：v） using two different soybean genotypes, Jack and Zhonghuang 10. The improved transformation efficiency of this combination was also evaluated by development of herbicide-tolerant soybeans with transformation efficiency at 2.5-5.7% for different geno- types, which was significantly higher than traditional cotyledonary node method in this study. These results suggested that co-treatment with surfactant and sonication significantly improved the percentages of resistance bud formation, transient expression efficiency and stable transformation efficiency in soybean.

Isolation of protein from Chlorella sorokiniana CY1 using liquid biphasic flotation assisted with sonication through sugaring-out effect

Microalgae,a sustainable source of multi beneficial components has been discovered and could be utilised in pharmaceutical,bioenergy and food applications.This study aims to investigate the sugaring-out effect on the recovery of protein from wet green microalga,Chlorella sorokiniana CY1 which was assisted with sonication.A comparison of monosaccharides and disaccharides as one of the phaseforming constituents shows that the monosaccharides,glucose was the most suitable sugar in forming the phases with acetonitrile to enhance the production of protein(52% of protein).The protein productivity of microalgae was found to be significantly influenced by the volume ratio of both phases,as the yield of protein increased to 77%.The interval time between the sonication as well as the sonication modes were influencing the protein productivity as well.The optimum protein productivity was obtained with 10s of resting time in between sonication.Pulse mode of sonication was suitable to break down the cell wall of microalgae compared to continuous mode as a lower protein yield was obtained with the application of continuous mode.The optimum condition for protein extraction were found as followed:200g/L glucose as bottom phase with volume ratio of 1:1.25,10s of resting time for ultrasonication,5s of ultrasonication in pulse mode and 0.25g of biomass weight.The high yield of protein about 81% could be obtained from microalgae which demonstrates the potential of this source and expected to play an important role in the future.

Extraction of agar from Eucheuma cottonii and Gelidium amansii seaweeds with sonication pretreatment using autoclaving method

The effect of sonication pretreatment condition on Eucheuma cottonii and Gelidium amansii seaweed towards agar extraction wae studied.Four parameters were changed during sonication to investigate the effects on agar yield and quality.These parameters include the time interval,concentration ratio,frequency,and intensity.The highest amount of agar extracted from Eucheuma cottonii species could be obtained from the time interval of 30min,seaweed weight to solvent volume ratio of 1:20,the frequency of 35Hz,and the sonication power intensity of 30%.For Gelidium amansii species,the best agar yield also could be obtained from the time interval of 30 min,1:20 of seaweed weight to water volume ratio,the frequency of 35Hz,and power intensity of 30%.From the experiment,sonication pretreatment signifi cantly influenced the yield and properties of extracted agar.The sonication with autoclaved seaweed produced agar containing less sulfate content,which is an excellent chemical property for gel electrophoresis applications.The gelstrength of sonication with autoclaving for both seaweeds,Eucheuma and Gelidium species was the highest among those by sonication with direct heating,which proved that sonication pretreatment with autoclaving could enhance the physical properties of the agar.

Effect of Mild Sonication Conditions on the Attributes of <i>Lactobacillus delbrueckii</i>ssp. <i>bulgaricus</i>LB-12

Lactobacillus delbrueckii ssp. bulgaricus is a widely used bacterium for the production of some fermented dairy products. Mild sonication intensity condition is a non-destructive technique that uses sound waves to cause cavitation in aqueous solutions and may improve the permeability of membranes, speed up the transfer of substrates and promote cellular growth and propagation. The objective was to determine the effect of mild sonication intensities at different temperatures on growth, bile tolerance and protease activity of Lactobacillus delbrueckii ssp. bulgaricus LB-12. The treatments were four sonication intensities (8.07, 14.68, 19.83 and 23.55 W/cm2) randomized at three different temperatures (4℃, 22℃ and 40℃). The energy input (1500 J) was kept constant in all treatments. Control samples did not receive any sonication treatment. Growth and bile tolerance were determined every 2 h for 12 h of incubation. Protease activity was determined at 0, 12 and 24 h. Mild sonication conditions included 1) mild sonication intensities, 2) temperatures and 3) times, all three of which played a role in influencing the desirable attributes of Lactobacillus delbrueckii ssp. bulgaricus LB-12. Of all the mild sonication intensities studied, 14.68 W/cm2 had the best overall influence at certain time points forimproving the bile tolerance and growth at 4℃ and protease activity at 40℃. Mild sonication intensity of 23.55 W/cm2 had the best overall influence at certain time points for protease activity of at 22℃. Some mild sonication conditions could be recommended for improvement of some characteristics of Lactobacillus delbrueckii ssp. bulgaricus LB-12.

Formation of graphene oxide from carbon rods of zinc-carbon battery wastes by audiosonic sonication assisted by commercial detergent

This study aims to determine the effect of audiosonic sonication in normal modes on the formation of graphene oxide(GO)fromcarbon rods of zinc-carbon(ZnC)battery wastes.The method used in this study was sonication with an audiosonic frequency in normal modes,assisted by a surfactant solution derived from a commercial detergent.A graphite-detergent solutionwas exposed to audiosonicwaves using a frequency of 170 Hz for 3 hwith a pattern on the surface of the solution.The graphite solution was a mixture of 0.8 g of graphite powder and 100 ml of distilled water that was mixed using a blender for 2 min.25 ml of the solution was then taken and dripped with two drops of detergent solution containing 0.2 g detergent powder dissolved into 100 ml distilled water,so that a graphite-detergent solutionwas obtained.The tools used in this study included UV–Visible spectroscopy(UV–Vis),Fourier TransformInfraRed spectroscopy(FTIR),and a Scanning Electron Microscope(SEM).The solution that was audiosonicated showed a strong visible nodal pattern on its surface.The UV–Vis spectroscopy produced absorbance peaks at wavelengths of 225 nmand 270 nm,and the FTIR indicated the presence of OH and C_C functional groups,which suggested the existence of GO.The SEMimages showed GO in the formof coral-like materials.

Effects of Sonication Processing on the Behavior of the Synthesis Human Serum Albumin-SPIONs Loaded PLGA Nanoparticles

This paper reports the most prominent contributions in the field of biodegradable polymeric nanoparticles from poly (lactic-co-glycolic acid) (PLGA) used as a protein/drug delivery. We use a combination of Human Serum Albumin (HSA)-superparamagnetic iron oxide nanoparticles (SPIONs) loaded PLGA nanoparticles. To obtain protein stabilization, the optimization of each step of synthesis nanoparticle is required. One of the most common problems in encapsulating protein to PLGA nanoparticles is the presence of several challenges as a problem of instability. We explained how the effect of the various sonication processing on the synthesis HSA-SPIONs loaded PLGA nanoparticles would be one of the crucial parameters for stability.

Influence of “Mild” Sonication Conditions on the Characteristics of <i>Streptococcus thermophilus</i>ST-M5

Mild sonication intensity is an acoustic energy which involves the conversion of electrical signal into a physical vibration modifying the permeability of the cell plasma membrane. The objective of this study was to determine the effect of mild sonication intensities at different temperatures on growth, bile tolerance and protease activity of Streptococcus thermophilus. The treatments were four mild sonication intensities (8.07, 14.68, 19.83 and 23.55 W/cm2) randomized at three different temperatures (4°C, 22°C and 40°C). The energy input (1500 J) was kept constant in all treatments. Control samples did not receive any sonication treatment. Growth and bile tolerance were determined every two hours for 12 h of incubation. Protease activity was determined at 0, 12 and 24 h. Mild sonication conditions included a) mild sonication intensities, b) temperatures and c) times, all three of which played a role in influencing the desirable attributes of Streptococcus salivarius ssp. thermophilus ST-M5. Of all the mild sonication intensities studied, 14.68 W/cm2 had the best overall influence at certain time points for improving bile tolerance and growth at 4°C, growth at 22°C and bile tolerance and growth at 40°C of Streptococcus salivarius ssp. thermophilus ST-M5. Mild sonication intensity of 23.55 W/cm2 had the overall best influence at certain time points for protease activity of Streptococcus salivarius ssp. thermophilus ST-M5 at 40°C. Streptococcus thermophilus ST-M5 pretreatment with some mild sonication conditions can be recommended for improvement of some of its characteristics.

Increased Detection of Polymicrobial Infections by Sonication and Significance of Prosthesis Design and Fixation Technique in Cases of Low-Grade PJI, a Retrospective Study

Introduction: Low-grade PJI remains a diagnostic dilemma in the medical community. It is mainly caused by low-virulent bacteria that cause chronic infection. This is mainly due to the formation of biofilms on the implant surface. This biofilm formation poses a diagnostic challenge that causes difficulties in the microbiological diagnosis of the infection by conventional culture methods. Sonication culture of implants has proven to be useful for biofilm forming bacteria. Therefore, we examined the evaluation of low-grade PJI using sonication culture methods and searched for influencing factors. The methods: Between October 2016 and November 2019 we retrospectively examined adult patients with suggestive clinical and laboratory findings of low-grade PJI. The patient's medical history, demographic information, prosthesis type, laboratory findings and length of hospital stay were collected. The results of the microbiological culture were divided into the results of the conventional preoperative and intraoperative culture method and the results of the sonication. In addition, laboratory chemical diagnostics of the joint punctures and CRP determination were performed. Additionally, the influence of prosthesis design and fixation technique, with and without cement, was investigated. The Results: Fifty-two patients with a mean age of 70.5 years were included in this study. The female population was more frequently affected. The most frequently affected joints were hip and knee. A higher percentage of pathogen detection was found at sonication with 60% of positive cultures, compared to 52% of intraoperative and 40% of preoperative conventional positive culture findings. The majority of patients were monomicrobial infections, with Staphylococcus epidermidis being the most frequently detected etiologic pathogen. The sonication culture was best able to detect a polymicrobial infection in 14% of cases compared to 6% of intraoperative and 4% of preoperative conventional cultures. Patients with a polymicrobial infection showed higher CRP values (P = 0.0172) and longer hospitalization (P = 0.0017) than monomicrobial infected patients. The results of the intraoperative conventional culture showed the highest congruence with the results of the sonication culture (concordance of 79%) compared to the preoperative culture. An infectious histopathological classification confirmed the microbiological ultrasound findings in 69% of cases with an infectious category. Cemented fixation showed a trend towards less positive microbiological findings, and total hip arthroplasty showed the highest detection of bacteria by sonication culture compared to bicondylar surface knee replacement (P = 0.0072). Conclusion: The sonication culture proved to be an important microbiological diagnostic tool, especially for the detection of polymicrobial infections in cases of low-grade PJI. This method has an established place in the diagnosis of low-grade PJI. The prosthesis design and the cemented fixation technique seem to have an influence on the detection of bacteria.

小型猪乳磨牙胚SHH的时空表达

目的本研究通过探索SHH在小型猪第三乳磨牙胚的帽状期、钟状期和分泌期的时空表达情况,拟阐明SHH在牙齿发育调控中的作用。方法收集小型孕猪第三乳磨牙胚,通过原位杂交、免疫组化和实时聚合酶链反应检测SHH的m RNA和蛋白表达水平。结果原位杂交及免疫组化可见SHH在帽状期少量表达于成釉器,但在釉结部位表达较强。在钟状期,SHH在间充质中表达明显增强;在分泌期,SHH在间充质和成釉器中都有所表达,尤其在成牙本质细胞和内釉上皮细胞中表达增强。PCR结果显示,在钟状期,SHH在上皮中表达较多,而在帽状期和分泌期,SHH在上皮和间充质中的表达未见明显统计学差异。结论SHH可能在调节上皮形态发育方面具有重要功能,参与调节信号级联,介导上皮和间充质之间的相互作用,本研究进一步完善了小型猪牙齿发育的信号分子网络。

miR-130a-5p靶向Runx2对牙槽骨成骨细胞增殖和分化的影响及其调控机制研究

目的:探究miR-130a-5p靶向Runt相关转录因子2(Runt-related transcription factor 2,Runx2)对牙槽骨成骨细胞(Alveolar osteoblast,AOB)增殖、凋亡、成骨分化的影响及其机制。方法:将AOB分为NC组、miR-NC组、miR-130a-5p组、miR-130a-5p+Runx2组和miR-130a-5p+SHH组。双荧光素酶报告验证miR-130a-5p对Runx2、Sonic hedgehog(SHH)的调控关系;CCK-8及EdU染色检测细胞增殖能力;AnnexinV-FITC/PI法检测细胞凋亡能力;ALP染色、茜素红染色检测细胞成骨分化能力;RT-qPCR检测miR-130a-5p、Runx2、SHH、神经胶质瘤关联癌基因同源物1(Gli1)mRNA水平;Western Blot检测增殖细胞核抗原(Proliferating cell nuclear antigen,PCNA)、Ki67、B细胞淋巴瘤/白血病-2(B-cell lymphoma/leukemia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 related X protein,Bax)、碱性磷酸酶(Alkaline phosphatase,ALP)、骨钙素(Osteocalcin,OCN)、骨形态发生蛋白2(Bone morphogenetic protein 2,BMP2)、Runx2、SHH、Gil1水平。结果:miR-130a-5p靶向调控Runx2、SHH。过表达miR-130a-5p后,细胞24 h、48 h、72 h OD值及EdU阳性率降低,细胞凋亡率升高,成骨分化能力降低,miR-130a-5p、Bax蛋白水平升高,Runx2、PCNA、Ki67、Bcl-2、ALP、OCN、BMP2蛋白及SHH、Gil1 mRNA和蛋白水平均降低(P<0.05)。过表达miR-130a-5p和Runx2或过表达miR-130a-5p和SHH可减弱过表达miR-130a-5p对细胞增殖、凋亡、成骨分化的影响。结论:miR-130a-5p靶向Runx2抑制AOB增殖和成骨分化,并促进AOB凋亡,其可能通过抑制SHH信号通路发挥作用。

Sonication and grinding pre-treatments on Gelidium amansii seaweed for the extraction and characterization of Agarose

Various prctreatments methods including sonication and grinding were perfonned on red seaweed Gelidium amansii for the subsequent extraction ofagarose. The agarose products are usually extracted from agar powder products from seaweeds. In this study, the agarose was extracted using a direct polyethylene glycol （PEG） method without the need to first process the agar from seaweed. The agar extract was frozen then thawed and mixed directly with PEG solution to precipitate the agarose. The quality of agarose obtained was evaluated through physico-chemical properties analysis which includes spectral technique （FTIR）, melting and boiling point, gel strength and sulfate content. These properties were compared with a non-pretreated sample and it was found that the addition of pretreatment steps improved the quality of agarosc but gave a slightly lower yield. The gel strength of pretreated samples was much higher and the sulfate content was lower compared to non-pretreated samples. The best pretreatment method was sonication which gave gel strength of 742 gcm - and sulfate content of 0.63%. The extraction of agarose can be further improved with the use of different neutralizing agents. Pretreating the seaweed shows potential in improving the quality of agarosc from seaweed and can be applied for thture extraction of the agarose.

Effect of Sonic hedgehog gene-modified bone marrow mesenchymal stem cells on graft-induced retinal gliosis and retinal ganglion cells survival in diabetic mice

AIM:To investigate the effects of Sonic hedgehog(Shh)gene-modified bone marrow mesenchymal stem cells(MSCs)on graft-induced retinal gliosis and retinal ganglion cells(RGCs)survival in diabetic mice.METHODS:Bone marrow-derived MSCs were genetically modified with the Shh gene to generate a stably transfected cell line of Shh-modified MSCs(MSC-Shh).Intravitreal injections of MSC-Shh and green fluorescent protein-modified MSCs(MSC-Gfp;control)were administered in diabetic mice.After 4wk,the effects of MSC-Shh on retinal gliosis were evaluated using fundus photography,and markers of gliosis were examined by immunofluorescence and Western blotting.The neurotrophic factors expression and RGCs survival in the host retina were evaluated using Western blotting and immunofluorescence.The mechanisms underlying the effects of MSC-Shh was investigated.RESULTS:A significant reduction of proliferative vitreoretinopathy(PVR)was observed after intravitreal injection of MSC-Shh compared to MSC-Gfp.Significant downregulation of glial fibrillary acidic protein(GFAP)was demonstrated in the host retina after MSC-Shh administration compared to MSC-Gfp.The extracellular signal-regulated kinase 1/2(ERK1/2),protein kinase B(AKT)and phosphatidylin-ositol-3-kinase(PI3K)pathways were significantly downregulated after MSC-Shh administration compared to MSC-Gfp.Brain-derived neurotrophic factor(BDNF)and ciliary neurotrophic factor(CNTF)levels were significantly increased in the host retina,and RGCs loss was significantly prevented after MSC-Shh administration.CONCLUSION:MSC-Shh administration reduces graft-induced reactive gliosis following intravitreal injection in diabetic mice.The ERK1/2,AKT and PI3K pathways are involved in this process.MSC-Shh also increases the levels of neurotrophic factors in the host retina and promoted RGCs survival in diabetic mice.

Hedgehog信号通路在下颌骨发育过程中作用机制的研究进展

下颌骨的发育来源及发育过程均与全身其他骨骼有着显著差异,其发育异常会导致各种骨相关疾病,严重影响了患者的生活质量。近年来Hedgehog信号通路在骨骼发育过程中的作用越来越受到关注。Hedgehog基因包括Sonic Hedgehog(Shh)、Indian Hedgehog(Ihh)和Desert Hedgehog(Dhh)3种亚型,其中Shh及Ihh可通过多种途径参与骨代谢调节,Shh主要参与肢体发育过程,而Ihh主要是在软骨内成骨过程中发挥关键作用。Hedgehog信号通路包括Hedgehog信号蛋白配体、Patched(Ptch)受体、Smoothed(Smo)受体、核内转录因子神经胶质瘤相关癌基因同源蛋白(glioma-associated oncogene homologue,Gli)和下游靶基因等。典型Hedgehog信号通路由Gli激活调控,而非典型Hedgehog信号主要由Ptch、Smo等调控。Shh在早期脊椎动物胚胎形成过程中调控多种生物学行为,如器官的分化、神经干的形成、干细胞的分化增殖、四肢骨骼发育以及牙胚发育等;在骨细胞分化过程中,Shh、Ptch1和Gli1在成骨细胞中均有表达,进一步促进骨髓间充质干细胞向成骨细胞和软骨细胞分化。Ihh对骨骼生长发育以及稳态维持具有不可或缺的功能作用,参与膜内骨领的形成、软骨细胞的增殖和成熟,Ihh在成熟的颅骨成骨细胞中表达,其可作为促成骨因子调控Ptch和骨形态发生蛋白(bone morphogenetic protein,BMP)的表达来诱导膜内骨化过程;脑和肌肉ARNT样蛋白1(brain and muscle ARNT-like 1,BMAL1)可通过与Ptch1和Ihh结合,调节Hedgehog信号通路,在颞下颌关节的软骨形成和软骨内成骨过程中发挥关键作用;而Hedgehog信号激活剂可以改善由BMAL1缺失引起的下颌骨骨量减少。Hedgehog信号失调会对骨发育过程产生重大影响,并导致一系列骨疾病如骨发育异常、骨折、骨质疏松和骨关节炎。然而,Hedgehog信号通路与下颌骨疾病的相关作用机制尚未完全阐明,未来研究可进一步探讨Hedgehog信号作为治疗下颌骨发育相关疾病的潜在靶点。

Frequency Effects on 2-Chlorobiphenyl Sonication

In order to better understand sonication, this paper studies the effect of sound frequency on the 2-chlorobiphenyl (2-CB) sonication and analyzes two prevailing hypotheses. Four frequencies 205, 358, 618, and 1071 kHz were tested. The 2-CB degradation kinetics, dechlorination, and inhibition by a free radical scavenger were examined. The results show that sonication effectively degrades 2-CB, and the first order rate constants using 0.4 W/cm3 sonication are 0.214, 0.508, 0.454, and 0.248 min1 at 205, 358, 618, and 1071 kHz, respectively. Good dechlorination was also achieved. 358 kHz frequency provided the most effi-cient 2-CB degradation, but the worst dechlorination, and was most sensitive to the free radical scavenger. Detailed analyses show that thermolysis is relatively stable at these frequencies while the free radical reac-tion depends strongly on the sound frequency.

Preconditioning process for dermal tissue decellularization using electroporation with sonication

Decellularization to produce bioscaffolds composed of the extracellular matrix(ECM)uses enzymatic,chemical and physical methods to remove antigens and cellular components from tissues.Effective decellularization methods depend on the characteristics of tissues,and in particular,tissues with dense,complex structure and abundant lipid content are difficult to completely decellularize.Our study enables future research on the development of methods and treatments for fabricating bioscaffolds via decellularization of complex and rigid skin tissues,which are not commonly considered for decellularization to date as their structural and functional characteristics could not be preserved after severe decellularization.In this study,decellularization of human dermal tissue was done by a combination of both chemical(0.05%trypsin-EDTA,2%SDS and 1%Triton X-100)and physical methods(electroporation and sonication).After decellularization,the content of DNA remaining in the tissue was quantitatively confirmed,and the structural change of the tissue and the retention and distribution of ECM components were evaluated through histological and histochemical analysis,respectively.Conditions of the chemical pretreatment that increase the efficiency of physical stimulation as well as decellularization,and conditions for electroporation and sonication without the use of detergents,unlike the methods performed in previous studies,were established to enable the complete decellularization of the skin tissue.The combinatorial decellularization treatment formed micropores in the lipid bilayers of the skin tissues while removing all cell and cellular residues without affecting the ECM properties.Therefore,this procedure can be widely used to fabricate bioscaffolds by decellularizing biological tissues with dense and complex structures.

M2 macrophages mediate fibrotic scar formation in the early stages after cerebral ischemia in rats

In the central nervous system, the formation of fibrotic scar after injury inhibits axon regeneration and promotes repair. However, the mechanism underlying fibrotic scar formation and regulation remains poorly understood. M2 macrophages regulate fibrotic scar formation after injury to the heart, lung, kidney, and central nervous system. However, it remains to be clarified whether and how M2 macrophages regulate fibrotic scar formation after cerebral ischemia injury. In this study, we found that, in a rat model of cerebral ischemia induced by middle cerebral artery occlusion/reperfusion, fibrosis and macrophage infiltration were apparent in the ischemic core in the early stage of injury(within 14 days of injury). The number of infiltrated macrophages was positively correlated with fibronectin expression. Depletion of circulating monocyte-derived macrophages attenuated fibrotic scar formation. Interleukin 4(IL4) expression was strongly enhanced in the ischemic cerebral tissues, and IL4-induced M2 macrophage polarization promoted fibrotic scar formation in the ischemic core. In addition, macrophage-conditioned medium directly promoted fibroblast proliferation and the production of extracellular matrix proteins in vitro. Further pharmacological and genetic analyses showed that sonic hedgehog secreted by M2 macrophages promoted fibrogenesis in vitro and in vivo, and that this process was mediated by secretion of the key fibrosis-associated regulatory proteins transforming growth factor beta 1 and matrix metalloproteinase 9. Furthermore, IL4-afforded functional restoration on angiogenesis, cell apoptosis, and infarct volume in the ischemic core of cerebral ischemia rats were markedly impaired by treatment with an sonic hedgehog signaling inhibitor, paralleling the extent of fibrosis. Taken together, our findings show that IL4/sonic hedgehog/transforming growth factor beta 1 signaling targeting macrophages regulates the formation of fibrotic scar and is a potential therapeutic target for ischemic stroke.

Anethole improves the developmental competence of porcine embryos by reducing oxidative stress via the sonic hedgehog signaling pathway

Background Anethole(AN)is an organic antioxidant compound with a benzene ring and is expected to have a positive impact on early embryogenesis in mammals.However,no study has examined the effect of AN on porcine embryonic development.Therefore,we investigated the effect of AN on the development of porcine embryos and the underlying mechanism.Results We cultured porcine in vitro-fertilized embryos in medium with AN(0,0.3,0.5,and 1 mg/mL)for 6 d.AN at 0.5 mg/mL significantly increased the blastocyst formation rate,trophectoderm cell number,and cellular survival rate compared to the control.AN-supplemented embryos exhibited significantly lower reactive oxygen species levels and higher glutathione levels than the control.Moreover,AN significantly improved the quantity of mitochondria and mitochondrial membrane potential,and increased the lipid droplet,fatty acid,and ATP levels.Interestingly,the levels of proteins and genes related to the sonic hedgehog(SHH)signaling pathway were significantly increased by AN.Conclusions These results revealed that AN improved the developmental competence of porcine preimplantation embryos by activating SHH signaling against oxidative stress and could be used for large-scale production of high-quality porcine embryos.

Sonic hedgehog信号通路Smo蛋白及其下游转录因子Gli1蛋白在胃癌组织中的表达及其意义

目的探讨Sonichedgehog信号通路Smo蛋白及其下游转录因子Gli1蛋白在胃癌组织中的表达及其意义。方法收集85例胃癌组织及25例正常胃粘膜组织构建成组织芯片,应用免疫组化S-P法检测胃癌组织中Smo蛋白及Gli1蛋白的表达,并以正常胃粘膜组织作对照,分析两者的相关性。结果Smo和Gli1蛋白均在正常胃粘膜中不表达或弱表达;在胃乳头状腺癌、管状腺癌、低分化腺癌中,两者的表达强度和阳性表达率显著高于正常胃粘膜(P<0.05,并随着胃腺癌分化程度下降而上升;相关分析显示Smo和Gli1蛋白在胃癌组织中的表达呈正相关,相关系数为0.989,P<0.001。结论胃癌发生过程中的Sonichedgehog信号通路异常激活可能通过Smo蛋白高表达上调下游转录因子Gli1蛋白的表达参与部分胃腺癌的发生。

Sonic hedgehog信号通路在单侧输尿管梗阻大鼠肾组织中的表达变化及意义

目的:探讨Sonic hedgehog(Shh)信号通路在单侧输尿管梗阻(UUO)大鼠肾组织中的表达变化及意义。方法:将48只SD大鼠随机分为UUO模型组(n=24)和假手术组(n=24),梗阻术3、7和14 d后取其梗阻侧肾脏组织。用HE和Masson染色检测肾间质纤维化程度,免疫组织化学染色检测Shh通路分子Shh、Ptch1、Smo、Gli1及Ⅲ型胶原的蛋白表达,酶联免疫吸附实验(ELISA)检测肾组织中TGF-β1和Shh含量,real-time RT-PCR检测TGF-β1、I和Ⅲ型胶原及Shh通路分子mRNA表达。结果:HE和Masson染色显示,梗阻侧肾组织出现明显的纤维化病变,且随时间延长而加剧。TGF-β1、I和Ⅲ型胶原含量在梗阻肾中表达明显增高(P<0.05)。同时,Shh信号通路分子Shh、Smo和Gli mRNA和蛋白在梗阻肾中表达明显升高(P<0.05),而Ptch1 mRNA和蛋白的表达下调(P<0.01),提示Shh信号被激活。相关分析表明,Shh信号起始信号Shh水平的升高与TGF-β1含量增加呈明显的相关。结论:UUO大鼠诱导肾间质纤维化发生过程中,Shh信号通路分子被激活,推测可能的机制是活化的Shh信号通路诱导TGF-β1表达和释放,导致肾间质纤维化。

Smo蛋白及Gli1蛋白在结直肠癌组织中的表达及其意义

目的探讨Sonic hedgehog信号通路中Smo和Gli1蛋白在结直肠癌组织中的表达及其意义。方法应用免疫组化S-P法检测125例结直肠癌组织及30例正常结直肠黏膜组织中Sonic hedgehog信号通路中Smo和Gli1蛋白的表达情况。结果Smo和Gli1蛋白在结直肠癌组织的表达明显高于其正常结直肠黏膜(P<0.001),与肿瘤部位、分化程度、Dukes分期以及有无淋巴结转移无关(P>0.05),而与肿瘤组织学类型有关(P<0.001),相关分析显示Smo及Gli1蛋白在结直肠癌组织中的表达呈正相关,相关系数为0.690(P<0.001)。结论结直肠癌的发生与Smo和Gli1蛋白的高表达有关,可能与Smo蛋白高表达上调其下游转录因子Gli1蛋白的表达有关。