张付峰医生的科普号

现代医学研究表明，菜籽油中含有40％的芥酸，而芥酸是一种长链脂肪酸，对心脏病人的心血管系统功能有很大危害。若心脏病人长期食用菜籽油，血液中每日吸收少许被酶消化了的芥酸后，就会使本来不正常的心血管功能超负荷，更容易诱发“血管壁增厚”及“心肌脂肪沉积”等病变，直接危害心脏病人的身体健康。为此，联合国粮农组织和世界卫生组织已对菜籽油中的芥酸含量做出限量规定，规定菜籽油中芥酸的含量一律不得超过5％。所以各类心脏病人，特别是冠心病、高血压、冠状动脉供血不足或心绞痛患者，应尽量少吃菜籽油。

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AbstractWe previously found that the K141N mutation in heat shock protein B8 (HSPB8) was responsiblefor Charcot-Marie-Tooth disease type 2L in a large Chinese family. The objective of thepresent study was to generate a transgenic mouse model bearing the K141N mutation in thehuman HSPB8 gene, and to determine whether this K141NHSPB8 transgenic mouse model wouldmanifest the clinical phenotype of Charcot-Marie-Tooth disease type 2L, and consequently besuitable for use in studies of disease pathogenesis. Transgenic mice overexpressing K141NHSPB8were generated using K141N mutant HSPB8 cDNA cloned into a pCAGGS plasmid driven by ahuman cytomegalovirus expression system. PCR and western blot analysis confirmed integrationof the K141NHSPB8 gene and widespread expression in tissues of the transgenic mice. TheK141NHSPB8 transgenic mice exhibited decreased muscle strength in the hind limbs and impairedmotor coordination, but no obvious sensory disturbance at 6 months of age by behavioral assessment.Electrophysiological analysis showed that the compound motor action potential amplitudein the sciatic nerve was significantly decreased, but motor nerve conduction velocity remainednormal at 6 months of age. Pathological analysis of the sciatic nerve showed reduced myelinatedfiber density, notable axonal edema and vacuolar degeneration in K141NHSPB8 transgenic mice,suggesting axonal involvement in the peripheral nerve damage in these animals. These findingsindicate that the K141NHSPB8 transgenic mouse successfully models Charcot-Marie-Tooth diseasetype 2L and can be used to study the pathogenesis of the disease.Key Words: nerve regeneration; peripheral nerve injury; axonal injury; animal models; Charcot-Marie-Tooth disease type 2L; gene mutation; pronuclear injection; transgenic model; small heat shockprotein B8; NSFC grant; neural regenerationIntroductionCharcot-Marie-Tooth disease is the most common inheritedperipheral neuropathy, with an estimated prevalence of 17–40/10,000[1-2]. Charcot-Marie-Tooth is characterized by distalmuscle weakness and atrophy, distal sensory loss, depressedtendon reflexes, and pes cavus. There are two main forms:the demyelinating type 1 (Charcot-Marie-Tooth disease type1) and the axonal type 2 (Charcot-Marie-Tooth disease type2). Autosomal-dominant inheritance is most common, butX-linked and autosomal-recessive forms are also seen[1-3].There is considerable genetic heterogeneity, and more than40 genes have been identified (http://neuromuscular.wustl.edu/time/hmsn.html); however, the underlying pathogeneticmechanisms remain unclear[4-5].Tang et al.[6] identified an autosomal dominant familialform of Charcot-Marie-Tooth disease type 2 in a Chinesefamily, linked to 12q24 by genome-wide screening, anddesignated Charcot-Marie-Tooth disease type 2L (OMIMNumber 608673). A positional candidate cloning studyshowed that the G423T (K141N) mutation in the smallheat shock protein B8 (HSPB8) gene co-segregated with theCharcot-Marie-Tooth disease type 2L phenotype, and that itwas the causative mutation[7]. The expression of K141NHSPB8in SHSY5Y cells results in intracellular aggregates that aremainly distributed in the cytoplasm and that are colocalizedwith HSPB1 and neurofilament light polypeptideNEFL[8-9].To provide insight into the molecular pathogenesis ofthe disease, in the present study, the transgenic plasmidpCAGGS-HA-K141NHSPB8 was constructed, and K141NHSPB8transgenic mice were generated by pronuclear injection.Behavioral, electrophysiological, and pathological analyseswere performed on K141NHSPB8 transgenic mice to deter-mine whether they manifest the clinical phenotype of Charcot-Marie-Tooth disease type 2L and would be suitable forstudies on disease pathogenesis.ResultsGeneration of K141NHSPB8 transgenic miceWe used PCR combined with sequencing for genotyping.The amplification yielded a 976-bp genomic fragment withprimers CAG1 and CAG2, and a 516-bp genomic fragmentwith primers CAG1 and HSP (Figure 1A–C). Sequencing ofthe 976-bp amplification product showed the presence ofthe G423T mutation when aligned with the HSPB8 sequence(Figure 1C). Western blot assay revealed extensive expressionof the HA-tagged 22-kDa human HSPB8 in heart andgastrocnemius muscle of founder mice (Figure 1D). Threefounder males were generated and bred normally to establishlines of K141NHSPB8 transgenic mice.Quantitative analysis of the experimental miceNine 6-month-old K141NHSPB8 transgenic mice, and nine agematchedC57BL mice, as control, were used for behavioralassessments, and data from all mice were included in the final analyses. Three of the nine 6-month-old K141NHSPB8 transgenicmice and three age-matched C57BL mice were used for sciaticnerve electrophysiological and histopathological studies.Impaired motor performance in K141NHSPB8 transgenicmiceAll K141NHSPB8 transgenic mice were normal at birth andshowed normal weaning and grooming behavior. The frequencyat birth of the various genotypes exhibited a normalMendelian inheritance pattern. At 6 months of age, themice displayed an unsteadiness of gait, reminiscent of the“steppage” gait in Charcot-Marie-Tooth disease patients.Tremor and seizures were absent. When walking over a flatsurface in a straight line, the transgenic mice moved clumsilyand displayed outward positioning of their hind pawwith a significantly more open hind paw angle than wildtype mice (P < 0.05; Figure 2A–C). In the fixed-bar test,all the wild type mice balanced and supported their bodyweight easily, and moved agilely on the bar. In contrast, thetransgenic mice either remained motionless or crawled withdifficulty using only their forelimbs, dragging their hindlimbs behind. Two of the transgenic mice dropped off thebar in all eight consecutive trials within 120–240 seconds. Inthe rotarod test, the K141NHSPB8 transgenic mice remained onthe rotating roller for a significantly shorter period of timethan wild type mice (P < 0.05; Figure 3). However, in thepain threshold experiment involving plantar electric shocks,there were no significant differences between K141NHSPB8mice and age-matched wild type mice.Electrophysiological alterations in the proximal and distalsciatic nerve in K141NHSPB8 transgenic miceK141NHSPB8 transgenic mice showed a significant decreasein the peak-to-peak amplitude of compound motor actionpotentials in both proximal and distal sciatic nerve at the ageof 6 months, confirming the axonal basis of the motor neuropathy(P < 0.01; Figure 4, Table 1). Motor nerve conductionvelocity was normal, indicating that the myelin sheathremained basically intact (Table 1).Histopathological alterations in the distal portion of thesciatic nerve in K141NHSPB8 transgenic miceSemithin sections of the distal portion of the sciatic nervefrom 6-month-old K141NHSPB8 transgenic mice revealed adecrease in the number of axons compared with wild typecontrol mice (P < 0.05; Figure 5A–C). No signs of demyelinationor remyelination were observed. Under the electronmicroscope, ultrathin sections of the sciatic nerve showedthe presence of notable axonal edema and degeneration, althoughthe myelin sheath remained relatively intact (Figure5D, E).DiscussionIn the present study, we created a transgenic mouse modelof Charcot-Marie-Tooth disease type 2L by overexpressinghuman HSPB8 containing the K141N mutation. The ubiquitous-expression pCAGGS plasmid, driven by the humancytomegalovirus expression system, has proven to be suitablefor the creation of transgenic mouse models. Therefore,we chose this plasmid for the generation of the K141NHSPB8transgenic mice[10-12]. PCR and western blot assays on theK141NHSPB8 transgenic founder mice confirmed that the humanG423T (K141N) mutant HSPB8 gene had successfullyintegrated into the mouse chromosome and was widely expressed.Footprint analysis, a behavioral assessment, mainly reflectsthe motor function status of the hind limbs. Becausethe outward positioning of the hind paw helps to controlthe forward momentum generated during walking, hindpaw angle was used as a specific index to evaluate the stateof muscle relaxation of the hind limbs[13-14]. The increasedangle of paw placement in K141NHSPB8 mice (6 months ofage, equivalent to human 16.1 years) suggested motor nerveimpairment in the hind limbs. In both the fixed-bar androtarod tests, the K141NHSPB8 mice showed notable impairmentin motor coordination. However, the pain thresholdexperiment involving plantar electric shocks did not showa significant difference between K141NHSPB8 mice and agematchedwild type mice. This suggests that sensory functionin K141NHSPB8 mice was not significantly affected.Electrophysiological study is mainly used to examine thephysiological status of large-diameter nerve fibers. Demyelinationof peripheral nerve results in a substantial reduction inconduction velocity, and other typical changes in motor nerveconduction, including a notable increase in motor nervelatency and conduction block. However, axonal deficit ischaracterized by notable decreases in the amplitude of compoundmotor action potentials[15-16]. The electrophysiologicalstudy of the peripheral nerve in K141NHSPB8 mice revealed asignificantly decreased amplitude of compound motor actionpotentials, and a relatively normal nerve conduction velocity.This suggests that the phenotype of K141NHSPB8 transgenicmice is caused by axonal deficits, rather than myelin abnormalities.Histopathological study of the peripheral nerve inK141NHSPB8 transgenic mice showed reduced numbers ofmyelinated fibers, axonal vacuoles and degeneration. Therewas no thickening of the myelin sheath or onion-bulb likestructures, which further suggests that peripheral neuropathyis an axonal neuropathy rather than a demyelinating one.The behavioral, electrophysiological and histopathologicalcharacteristics of the transgenic mice are similar to the featuresin patients with familial Charcot-Marie-Tooth diseasetype 2L, in whom the onset of disease occurs at young ormiddle age. In these patients, impairment mainly involvesthe lower limbs, and electrophysiological and histopathologicalstudies show an axonal neuropathy.The G423T (K141N) mutation in HSPB8 causes Charcot-Marie-Tooth disease type 2L. Interestingly, the K141Nand K141E mutations in HSPB8 were also identified in familieswith distal hereditary motor neuropathy (dHMN)[17-18].The dHMNs comprise a heterogeneous group of diseasesthat share the common feature of a length-dependent predominantmotor neuropathy. Many forms of dHMN haveminor sensory abnormalities and/or a significant uppermotor neuron component, and there is often an overlapwith the axonal forms of Charcot-Marie-Tooth disease type2 and with juvenile forms of amyotrophic lateral sclerosisand hereditary spastic paraplegia[19]. Patients in the Charcot-Marie-Tooth disease type 2L family had mild sensoryimpairments, including deficits in pain and touch, but noevidence of painless injury or ulceration. While both startin adulthood, distal HMN type II progresses more rapidly,and complete paralysis of all distal muscles of the lower extremitiesoccurs within 5 years, while Charcot-Marie-Toothdisease type 2L progresses slowly and all patients remainambulant. It remains unclear how the K141N mutation inHSPB8 produces different phenotypes in the two differentdiseases. In the present study, the K141NHSPB8 transgenicmice developed slowly progressing hind limb weakness, andhad no obvious sensory impairment at the age of 6 months,indicating relatively mild disease progression, with little orno sensory disturbance.We consider the K141NHSPB8 transgenic mice that wegenerated to be a suitable model of human Charcot-Marie-Tooth disease type 2L. However, the degree of sensoryinvolvement in the peripheral nerves needs to be furtherinvestigated. Further study on dorsal root ganglion neuronsis needed to clarify whether the sensory system is affected tovarying degrees in individual transgenic mice[20-21].In a previous study, combined expression of HSPB8K141N and HSPB8 K141E mutant proteins in motor neuronsresulted in neurite degeneration, manifested by areduction in number of neurites per cell and a reductionin average length of neurites[22]. In early passage, primary fibroblastcultures derived from dHMN patient skin biopsies,HSPB8 protein aggregates were present and mitochondrialmembrane potential was reduced[23]. Cytoplasmic aggregateswere observed when K141NHSPB8 was transiently expressed incultured cells, and cell viability was impaired after heat shocktreatment[24]. Overexpression of mutant HSPB8 was found toresult in autophagosomes that colocalized with protein aggregates,but failed to colocalize with lysosomes[25]. HSPB8 andBag3 form a chaperone complex that stimulates degradationof protein substrates by macroautophagy. Thus, defects inHSPB8-mediated autophagy are likely to be pathogenic[26-30].We anticipate that the novel K141NHSPB8 transgenic mousemodel, coupled with cell and tissue culture systems, will bea valuable research tool for elucidating the cellular and molecularpathogenesis of Charcot-Marie-Tooth disease type 2,and should advance the development of novel therapeuticstrategies for this neurological disorder.Materials and MethodsDesignEstablishment of a transgenic mouse model.Time and settingMajor components of experiments were performed atthe State Key Laboratory of Medical Genetics and ThirdXiangya Hospital, Central South University, China; parts ofexperiments were performed at Institute of Neuroscience,Shanghai Institute for Biological Sciences and Kunming Instituteof Zoology, Chinese Academy of Sciences, China fromMarch 2008 to August 2012.MaterialsA total of nine clean K41NHSPB8 transgenic mice (four females,five males), aged 6 months, weighing 23–28 g, and nineclean wild type C57BL mice (four females, five males), aged 6months, weighing 23–28 g, as controls, were provided by theInstitute of Laboratory Animal Sciences, Chinese Academyof Medical Sciences in China (license No. SCXK (Jing) 2005-0013). Animals were housed at the Laboratory in the AnimalFacility of Third Xiangya Hospital, Central South Universityin China. Animals were reared in 17 cm × 26 cm cages, eachcontaining two or three mice, under a 12-hour light/darkcycle (lights off between 18:00–06:00), at an average temperatureof 22°C, with free access to food and water. This studywas approved by the Animal Ethics Committee, Third XiangyaHospital, Central South University, China.MethodsCreation and genotyping of K141NHSPB8 transgenic miceThe pEGFPN1-K141NHSPB8 vector constructed in our previousstudy was used as a template to clone HA-taggedK141NHSPB8 cDNA using the EcoRI restriction endonucleaseinto the pCAGGS plasmid[8]. An expression vector driven bya human cytomegalovirus immediate-early enhancer linkedto the chicken β-actin promoter was used[8]. We excised a2,896-bp fragment from the pCAGGS-HA-K141NHSPB8 vectorwith the restriction endonucleases SalI and HindIII, andthen with BsaXI. We purified the fragment from an agarosegel with the QIAquick Gel Extraction Kit (Qiagen, Hilden,Germany), dialyzed it against injection buffer, and diluted itto a concentration of 2 ng/μL. After preparing C57BL femalemice for ovulation and egg fertilization, a 0.5-μL aliquot ofDNA was microinjected into the fertilized eggs. Injected zygoteswere maintained overnight and transferred into pseudo-pregnant C57BL females. The mice were placed in individuallyventilated cages. The tail tips of 1-week-old pupswere collected (1–2 cm/pup), and the phenol-chloroformmethod was used to extract DNA. Genotyping of transgenicanimals was performed with two primer sets: forwardprimer (CAG1), 5′-GCC ACC ATG TAC CCA TAC G-3′ andreverse primer (CAG2), 5′-GCA GGA GGC TGT TTC ATA-3′ to amplify the transgenic construct including the wholeHSPB8 gene; forward primer CAG1 and reverse primer(HSP), 5′-TGG GGA AAG TGA GGC AAA TA-3′ to amplifythe transgenic construct including part of the HSPB8 geneto confirm that HSPB8 had inserted into the mouse genome.PCR products were separated by 8% polyacrylamide gelelectrophoresis, and DNA sequencing was performed usingan ABI PRISM 3100 sequencer (Perkin Elmer, Waltham, MA,USA). Sequencing results were analyzed with DNASTAR Lasergene.v7.1 software (DNASTAR. Inc, Madison, WI, USA).The transgenic founders were identified and subsequentlytransferred to the animal facility of Third Xiangya Hospital,Central South University in China.Western blot assay for HA-tagged HSPB8 in K141NHSPB8founder miceThree 10-month-old K141NHSPB8 founder mice and onewild type C57BL control were killed by cervical dislocation,and flash-frozen tissue samples were maintained at –80 °C.Frozen tissues from heart and gastrocnemius muscles werehomogenized in radio immunoprecipitation assay buffer(containing 50 mmol/L Tris, 150 mmol/L NaCl, 1% TritonX-100, 1% sodium deoxycholate, 0.1% sodium dodecylsulfate and phosphatase inhibitors). Protein concentrationswere determined using the microBCA kit (Pierce ChemicalCo., Rockford, IL, USA) according to the manufacturer’sinstructions. Western blot assay was performed as describedbefore[8]. Goat anti-HSPB8 antibody (1:500; Sigma-Aldrich,Milwaukee, WI, USA), mouse anti-HA-Tag antibody(1:2,000; Sigma-Aldrich) and mouse anti-β-actin antibody(1:500; Sigma-Aldrich) were used as primary antibodiesand incubated with the blots at 4°C for 2 hours. Rabbit anti-goat IgG (1:2,000; Sigma-Aldrich) and goat anti-mouselgG (1:6,000 or 1:10,000; Sigma-Aldrich) were the secondaryantibodies, and were incubated with the blots at 4°C for 2hours. The target bands were visualized using an enhancedchemiluminescence detection kit (BioRad, Hercules, CA,USA) and then exposed to X-ray film in a dark room. TheX-ray film signal was scanned on an imaging analysis system(Bio-Rad), and absorbance values were analyzed.Behavioral assessments of K141NHSPB8 transgenic miceFor each behavioral experiment, each mouse was testedtwice a day for 4 days in a row to ensure the repeatabilityand reliability of the data. Footprint analysis was performedas described by Sereda et al.[13]. Waterproof black ink wasapplied to the hind paws and footprints were taken on watercolorpaper. The animal was allowed to walk forward in a12-cm-wide lane. The footprints were then scanned and theangle of deviation of the median foot axis with regard to themovement axis (i.e., the hind paw angle) was measured for10 clearly visible footprints per mouse.Fixed-bar test was performed as described by Norreel etal.[31-32]. For the fixed-bar test, a round wooden bar (diameter:1.5 cm; length: 50 cm) was positioned 40 cm abovethe cage floor. Mice were placed on the middle of the bar.The movements and the time (in seconds) that the animalsremained on the bar were monitored using videos. A maximumof 5 minutes per trial was allowed. Motor coordinationwas further assessed with the rotarod test, as describedby Zhao et al.[33]. The apparatus (DXP-2, Institute of MateriaMedica, Chinese Academy of Medical Sciences, Beijing,China), consisting of a base platform and a rotating rodwith a diameter of 2.5 cm, was subdivided into four equalsections. The mice were trained to reach their performancebaseline with 5 sessions of 2-minute periods of walking at4 r/min per day for two days before the test. After training,each mouse was placed on the rod and allowed to rest for30 seconds, and then the rod was rotated at 2 r/min. Therotation speed was increased every 30 seconds to 4, 8, 12,16, 24, 28, 32 and 36 r/min. In a series of six trials per animal,the time (in seconds) that the mouse remained on therod was measured. Pain threshold test was performed aspreviously described[34-35]. Mice were placed in a ThresholdActivity Monitoring system (Med Associates Inc., Pittsburgh,PA, USA). Electric shocks began at 0.2 mA, 0.5 seconds, andwere increased by 0.1 mA in 1-min intervals until the mousehad the first pain response (either limb flick or jump). Theamplitude of the current at which the mouse had the firstpain response was recorded.Nerve electrophysiology in K141NHSPB8 transgenic micesFollowing anesthesia with pentobarbital (240 mg/kg), micewere fixed on a thermostat-controlled heating plate at 37°C.The sciatic nerve on one side was carefully isolated. The sciaticand posterior tibial nerves were stimulated with needleelectrodes inserted alongside the nerves at the sciatic notchand at the hock. The distance between the two points was1 cm. The square pulses (1 Hz, 3 mV) were delivered fivetimes per site using the BL-420E+ biological and functionalexperimental system (Pclab, Chengdu, China). Compoundmotor action potentials were recorded from an intrinsic footmuscle using a concentric needle electrode. The latencies ofcompound motor action potentials, elicited by stimulation atboth proximal (hip) and distal (hock) sites, were measuredfrom the stimulus artifact to the first negative deflection.Amplitudes were determined as the maximum peak-to-peakvoltage. Motor nerve conduction velocity was calculatedusing the distance between the two points of stimulation(1 cm) and the difference in the latencies between the twopoints (conduction time).Sciatic nerve histopathology in K141NHSPB8 transgenic miceAfter nerve electrophysiology, anesthetized mice were perfusedthrough the left ventricle with saline followed by 4%paraformaldehyde in cacodylate buffer (pH 7.2). The sciaticnerves on the other side were carefully isolated and dissectedfrom the region of the sciatic notch to the hock. Nerve specimenswere placed in either 4% paraformaldehyde (0.05 mol/LPBS, pH 7.2) or 2.5% glutaraldehyde. Specimens fixed in 4%paraformaldehyde were embedded in paraffin, subjected tosemi-thin (1-μm) sectioning, stained with 1% toluidine blueat 80°C for 30–45 seconds, then observed with an OlympusCX31 light microscope (Olympus Corporation, Tokyo, Japan).The number of axons was counted on four slides fromeach mouse. Specimens in 2.5% glutaraldehyde were embeddedin porous rubber bodies, subjected to ultrathin (50-nm)sectioning, double stained with uranyl acetate and lead nitrate,and then observed with an H-7500 transmission electronmicroscope (Hitachi, Tokyo, Japan) and photographed.Statistical analysisData were expressed as mean ± SEM, and processed withSPSS 19.0 software (SPSS, Chicago, IL, USA). Differencesbetween repeated measures of different genotypes werecompared using two-way analysis of variance followed byBonferroni’s post hoc test. Comparisons of means were madeusing a two-sample t-test. A value of P < 0.05 was consideredstatistically significant.Author contributions: Zhang RX and Zhang FF participated inthe study conception and design. Zhang FF, Li XB, Huang SX, ZiXH, Liu T, Liu SM and Li XN provided the data and ensured itsintegrity. Zhang RX and Zhang FF analyzed the data. Zhang RXwrote the manuscript and was in charge of manuscript revision.Zhang RX and Zhang FF obtained funding. Xia K, Pan Q andTang BS provided technical support. All authors approved the finalversion of this paper.Conflicts of interest: None declared.Peer review: Although progress in molecular genetics research issignificant, the pathological mechanism of Charcot-Marie-Toothstill waits to be unveiled. In this study, the K141NHSPB8 transgenicmouse model is successfully generated, and can be used as a modelto study the pathogenesis of Charcot-Marie-Tooth disease type 2L.References[1] Dyck PJ, Thomas PK. Peripheral Neuropathy. 4th ed. Philadelphia:Elsevier Saunders. 2005.[2] Patzkó A, Shy ME. 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猝死，从佛家来说，是一个人修来的福气，但对于凡夫俗子来说，却是对整个家族带来了灾难性的后果。一个个令人惋惜的案例仍清晰地不时闪现在我们的脑海，胡可心，38岁，青年科学家，心脏病突发猝死；高秀敏，46岁，著名演员，突发心梗猝死；王均瑶，38岁，，均瑶集团董事长，长期劳累与高压致肠癌；吴立君，36岁，御泥坊创始人，长期熬夜致脑疾发作猝死；李开复，51岁，长期熬夜致淋巴癌。 从健康管理理论看，一个人的一生动态波动于“健康”、“亚健康”、“疾病”三种状态之中，一个群体也可以根据健康状态分为“健康”、“亚健康”、“疾病”三种人群。个体从健康到疾病也需要一个经历一完整发生发展过程，是从低危险状态到高危险状态，再到发生早期改变，出现临床症状。中域康达倡导的健康管理服务理念和提供的健康管理服务，正是对这三种不同的健康状态和相应人群，通过健康教育、健康干预和健康促进等不同手段，维护和恢复其至健康状态。

感悟是对生活中真善美的认同和信任，是对自己心灵杂质的摈弃和净化不论你遇见谁，他都是对的不论发生什么事，那都是唯一发生的事不管事情开始于哪个时刻，那都是对的时刻已经结束的，就让他结束吧当生命中有些事情结束，他会帮我们进化要完整享受已然发生的事情，最好是放下并继续前行善待自己，爱你的存在让自己健康快乐地生活让自己自由开心地生活放下，自在，念佛阿弥陀佛

宁静才能致远平心才能静气静气才能干事干事才能成事涵养静气过程追求内在平衡营造一种和谐积蓄一种底蕴成就一种境界

脑动脉硬化中药妙方-活血通脉汤中南大学湘雅医院老年病科 张付峰丹参 30g三七粉 3g赤芍 10g葛根 20g制何首乌 20g酒女贞子 20g法半夏 6g每日一剂，十剂为一疗程此方功能主治：活血化瘀，益气养血， 滋肾豁痰。

（化淤解瘫散）中南大学湘雅医院 张付峰黄芪 10g葛根 10g川穹 6g益母草 15g毛冬青 20g丹参 10g红花 6g广地龙 10g陈皮 6g天麻 6g秦麔 10g连翘 10g石菖蒲 6g此方功效：补气活血，化淤消栓宁络。

便秘是指由于粪便在肠内停留过久，以致大便次数减少、大便干结、排出困难或不尽。一般两天以上无排便，可提示便秘存在。如果每天均排大便，但排便困难且排便后仍有残便感，或伴有腹胀，也应纳入便秘的范围。 便秘时，常出现下腹膨胀，便意未尽，严重者出现食欲不振、头昏、无力等症状，这可能与粪便的局部机械作用引起神经反射有关。 对有便秘情况的患者建议如下： (一)便秘预防 因为粪便主要是由食物消化后构成的，所以通过饮食调节来防治大便秘结是简单易行的方法。首先要注意饮食的量，只有足够的量，才足以刺激肠蠕动，使粪便正常通行和排出体外。特别是早饭要吃饱。其次要注意饮食的质，主食不要太精过细，要注意吃些粗粮和杂粮，因为粗粮、杂粮消化后残渣多，可以增加对肠管的刺激量，利于大便运行。副食要注意多食含纤维素多的蔬菜，因为正常人每公斤体重需要90-100毫克纤维素来维持正常排便。可多食青菜、韭菜、芹菜、蕃芋等。因为纤维索不易被消化吸收，残渣量多，可增加肠管内的容积，提高肠管内压力，增加肠蠕动，有利于排便。还有就是要多喝水，特别是重体力劳动者，因出汗多，呼吸量大，水分消耗多，肠管内水份必然被大量吸收，所以要预防大便干燥就得多喝水。早饭前或起床后喝一杯水有轻度通便作用。足量饮水，使肠道得到充足的水分可利于肠内容物的通过。另外可有意多食含脂肪多的食品，如核桃仁、花生米、芝麻、菜籽油、花生油等，它们都有良好的通便作用。(二)养成良好的排便习惯每个人都有各种习惯，大便也不例外，到一定的时间就要排便，如果经常拖延大便时间，破坏良好的排便习惯，可使排便反射减弱，引起便秘，所以不要人为地控制排便感。对经常容易发生便秘者一定要注意把大便安排在合理时间，每到时间就去上厕所，养成一个良好的排便习惯。(三)积极锻炼身体活动、活动，大便自通。散步、跑步，作深呼吸运动、练气功、打太极拳，转腰抬腿、参加文体活动和体力劳动等可使胃肠活动加强、食欲增加，膈肌、腹肌、肛门肌得到锻炼；提高排便动力，预防便秘。经常劳动的农村老年人很少便秘，而懒于活动，养尊处优的城市老年人便秘者较多，说明了这个道理。古代曾用导引术来防治便秘。《杂病源流犀浊》曰“保生秘要曰，以舌顶上腭，守悬壅，静念而液自生，俟满口，赤龙搅动，频漱频吞，听降直下丹田，又守静咽数回，大肠自润，行后功效。”对于年高体弱之人实为相宜。(四)及时治疗有关疾病有关疾病的治疗对预防大便秘结亦有一定的作用。如过敏性结肠炎、大肠憩室炎、结肠肿瘤、结肠狭窄；甲状腺功能低下、糖尿病；子宫肌瘤；铅、汞等金属中毒。另外可用中药同米煮粥吃来预防便秘：1.红薯粥 将红薯半斤洗净连皮切成小块，与粳米2-3两，加水适量煮粥，待粥成时，加白糖适量再煮二、三沸即可。2.紫苏麻仁粥 取紫苏子、麻子仁各10-15克，捣烂如泥，加水慢研，滤汁去渣，再用粳米2两煮为稀粥食用。老年或产后服用较为适宜。3.柏子仁粥 柏子仁10-15克，去皮捣烂，加粳米1-2两，水适量，煮粥。