Myelin histology:a key tool in nervous system research

The myelin sheath is a lipoprotein-rich,multilayered structure capable of increasing conduction velocity in central and peripheral myelinated nerve fibers.Due to the complex structure and composition of myelin,various histological techniques have been developed over the centuries to evaluate myelin under normal,pathological or experimental conditions.Today,methods to assess myelin integrity or content are key tools in both clinical diagnosis and neuroscience research.In this review,we provide an updated summary of the composition and structure of the myelin sheath and discuss some histological procedures,from tissue fixation and processing techniques to the most used and practical myelin histological staining methods.Considering the lipoprotein nature of myelin,the main features and technical details of the different available methods that can be used to evaluate the lipid or protein components of myelin are described,as well as the precise ultrastructural techniques.

Time-dependent impact of a high-fat diet on the intestinal barrier of male mice

BACKGROUND Excessive saturated fat intake compromises the integrity of the intestinal mucosa,leading to low-grade inflammation,impaired mucosal integrity,and increased intestinal permeability,resulting in the migration of lipopolysaccharide(LPS)to other tissues.AIM To evaluate the chronic effects(at 10 and 16 wk)of a high-fat diet(HFD)(with 50%energy as fat)on the phylogenetic gut microbiota distribution and intestinal barrier structure and protection in C57BL/6 mice.METHODS Forty adult male mice were divided into four nutritional groups,where the letters refer to the type of diet(control and HFD or HF)and the numbers refer to the period(in weeks)of diet administration:Control diet for 10 wk,HFD for 10 wk,control diet for 16 wk,and HFD for 16 wk.After sacrifice,biochemical,molecular,and stereological analyses were performed.RESULTS The HF groups were overweight,had gut dysbiosis,had a progressive decrease in occludin immunostaining,and had increased LPS concentrations.Dietary progression reduced the number of goblet cells per large intestine area and Mucin2 expression in the HF16 group,consistent with a completely disarranged intestinal ultrastructure after 16 wk of HFD intake.CONCLUSION Chronic HFD intake causes overweight,gut dysbiosis,and morphological and functional alterations of the intestinal barrier after 10 or 16 wk.Time-dependent reductions in goblet cell numerical density and mucus production have emerged as targets for countering obesity-driven intestinal damage.

Structural Features of Nuclei in Leaf Mesophyll Cells of Salt-Tolerant Artemisia marschalliana

All nuclei in mesophyll cells of Artemisia marschalliana are located in vacuoles and occupy up to 90% of their volume. The ultrastructural organization of chromatin in nuclei shows different degrees of its decondensation, up to complete separation of DNA from histones. It is possible that the separation of DNA from histones enables Artemisia to grow in soils with high salinity.

Effects of tachyplesin on the morphology and ultrastructure of human gastric carcinoma cell line BGC-823

AIM To investigate the morphological andultrastructural changes in the human gastriccarcinoma cell line BGC-823 after being treatedwith tachyplesin.METHODS Tachyplesin was isolated from acidextracts of Chinese horseshoe crab(Tachypleustridentatus)hemocytes.BGC-823 cells and thecells treated with 2.0mg/L tachyplesin wereexamined respectively under light microscope,scanning and transmission electron microscope.RESULTS BGC-823 cells had undergone therestorational alteration in morphology andultrastructure after tachyplesin treatment.Thechanges were as follows:the shape of cells wasunanimous,the volume enlarged and cellsturned to be flat and spread,the nucleo-cytoplasmic ratio lessened and nuclear shapebecame rather regular,the number of nucleolusreduced and its volume lessened,heter-chromatin decreased while euchromatinincreased in nucleus.In the cytoplasm,mitochondria grew in number with consistentstructure relatively,Golgi complex turned to betypical and well-developed,rough endoplasmicreticulum increased and polyribosomedecreased.The microvilli at cellular surfacewere rare and the filopodia reduced whilelamellipodia increased at the cell edge.CONCLUSION Tachyplesin could alter themalignant morphological and ultrastructuralcharacteristics of human gastric carcinoma cellseffectively and have a certain inducing differen-tiation effect on human gastric carcinoma cells.

Ultrastructure of neuronal-like cells differentiated from adult adipose-derived stromal cells

β-mercaptoethanol induces in vitro adult adipose-derived stromal cells （ADSCs） to differentiate into neurons. However, the ultrastructural features of the differentiated neuronal-like cells remain unknown. In the present study, inverted phase contrast microscopy was utilized to observe β-mercaptoethanol-induced differentiation of neuronal-like cells from human ADSCs, and immunocytochemistry and real-time polymerase chain reaction were employed to detect expression of a neural stem cells marker （nestin）, a neuronal marker （neuron-specific enolase）, and a glial marker （glial fibrillary acidic protein）. In addition, ultrastructure of neuronal-like cells was observed by transmission election microscopy. Results revealed highest expression rate of nestin and neuron-specific enolase at 3 and 5 hours following induced differentiation; cells in the 5-hour induction group exhibited a neuronal-specific structure, i.e., Nissl bodies. However, when induction solution was replaced by complete culture medium after 8-hour induction, the differentiated cells reverted to the fibroblast-like morphology from day 1. These results demonstrate that β-mercaptoethanol-induced ADSCs induced differentiation into neural stem cells, followed by morphology of neuronal-like cells. However, this differentiation state was not stable.

Changes in aortic endothelium ultrastructure in male rats following castration, replacement with testosterone and administration of 50α-reductase inhibitor

Aim： To investigate the relationship between low androgen level and ultrastructure of vascular endothelium. Methods： Forty-eight male Sprague-Dawley rats were randomly divided into four groups： group A, normal rats with sham castration; group B, castrated rats; group C, castrated rats given testosterone （T） undecanoate; and group D, intact rats treated with 5α-reductase inhibitor. After 10 weeks of treatment or castration, rats in different groups were killed and serum T, free T （FT） and dihydrotestosterone （DHT） were measured. The aortic endothelia were scanned under electron microcopy and the Vascular Endothelium Structure Score （VESS） was computed. Results： Serum T and FT concentrations of rats in group B were significantly lower than those of the other three groups （P 〈 0.01）; DHT concentrations of group D rats were significantly decreased （P 〈 0.01 ） when compared with those of groups A and C. Rats in groups B and D rats （with low androgen levels） had obvious damage to their endothelial surfaces, which appeared crimpled, rough, adhesive and ruptured, and had high destruction of VESS. Conclusion： These results suggest that low concentrations of T and DHT are associated with ultrastructural damage of the aortic endothelia in male rats.

Influence of lanthanum on chloroplast ultrastructure of soybean leaves under ultraviolet-B stress

In order to investigate the effects of lanthanum（Ⅲ） on cell ultrastructure of soybean leaves under elevated ultraviolet-B irradiation （UV-B, 280-320 nm）, the chloroplast ultrastructure of soybean seedlings was studied by hydroponics under laboratory conditions. The resuits showed that the thylakoid in chloroplast was orderly and clearly as soybean leaves were pretreated by La（Ⅲ）. The thylakoid was indis- tinctly disordered, expanded and even indiscoverable in the chloroplast under UV-B stress. The impact on the thylakoid by the high intensity UV-B irradiation （T2） was bigger than that by the low intensity UV-B irradiation （T1）. However, the destruction of the chloroplast structure caused by UV-B stress was alleviated by La（Ⅲ）, and the arrangement of the thylakoid in the chloroplast became orderly and clearly. The effect of the alleviation by La（Ⅲ） under the low intensity UV-B irradiation （T1） was better than that under the high intensity UV-B irradiation （T2）.

Effects of potassium deficiency on photosynthesis,chloroplast ultrastructure,ROS,and antioxidant activities in maize(Zea mays L.)

Potassium(K)deficiency significantly decreases photosynthesis due to leaf chlorosis induced by accumulation of reactive oxygen species(ROS).But,the physiological mechanism for adjusting antioxidative defense system to protect leaf function in maize(Zea mays L.)is unknown.In the present study,four maize inbred lines(K-tolerant,90-21-3 and 099;K-sensitive,D937 and 835)were used to analyze leaf photosynthesis,anatomical structure,chloroplast ultrastructure,ROS,and antioxidant activities.The results showed that the chlorophyll content,net photosynthetic rate(P_n),stomatal conductance(G_s),photochemical quenching(q_P),and electron transport rate of PSII(ETR)in 90-21-3 and 099 were higher than those in D937 and 835 under K deficiency treatment.Parameters of leaf anatomical structure in D937 that were significantly changed under K deficiency treatment include smaller thickness of leaf,lower epidermis cells,and vascular bundle area,whereas the vascular bundle area,xylem vessel number,and area in 90-21-3 were significantly larger or higher.D937 also had seriously damaged chloroplasts and PSII reaction centers along with increased superoxide anion(O_2^-·)and hydrogen peroxide(H_2O_2).Activities of antioxidants,like superoxide dismutase(SOD),catalase(CAT),and ascorbate peroxidase(APX),were significantly stimulated in 90-21-3 resulting in lower levels of O_2^-·and H_2O_2.These results indicated that the K-tolerant maize promoted antioxidant enzyme activities to maintain ROS homeostasis and suffered less oxidative damage on the photosynthetic apparatus,thereby maintaining regular photosynthesis under K deficiency stress.

Expression of neurocan mRNA and ultrastructure of brain tissue after cerebral ischemia and reperfusion in stroke-prone renovascular hypertensive rats treated by electroacupuncture

We established a stroke-prone renovascular hypertensive rat model by bilateral constriction of the renal artery with sliver loop clips. After ten weeks, middle cerebral artery occlusion was induced for 2 hours. The rats then received electro-acupuncture at Baihui （DU 20） and Dazhui （DU 14） after onset of ischemia for 30 days. In situ hybridization study showed that electroacupuncture significantly reduced the number of neurocan mRNA-positive cells in the ischemic penumbra and hippocampal tissues of rats. Electron microscopy results demonstrated that the structure of neurons and blood vessels in the ischemic tissues were restored with electroacupuncture. Overall, these data suggest that electroacupuncture may protect neurons against ischemic reperfusion injury in stroke-prone renovascular hypertensive rats, which may be regulated by downregulation of expression of nerve inhibitory factor neurocan mRNA.

Photosynthetic Characteristics and Ultrastructure of Chloroplast of Cucumber Under Low Light Density in Solar-Greenhouse

The photosynthetic characteristics and ultrastructure of chloroplast of cucumber in solar-greenhouse were studied. The result showed that the photosynthetic rate (Pn), photosyntheticability (A350), carboxylation efficiency, light saturation point and light compensation pointall declined remarkably under low light density, indicating that the photosynthetic characteristicsof cucumber were closely related to light environment. Under low light density, the minimalfluorescence (Fo), alterable fluorescence (Fv), photochemical efficiency of PSⅡ(Fv/Fm), steadyfluorescence in light (Fs), maximal fluorescence (Fm) and actual efficiency of PSⅡ(φPSⅡ)etcincreased, indicating that the photochemical activity and efficiency for solar energy transformationenhanced, thus the light proportion used to electron transport also increased. The chlorophylla, b, a/b and carotenoid of shading leaves decreased. However, the depressed extent of Chl a andChl a/b were obviously larger than that of Chl b. The number of chloroplast and starch grainin cucumber leaves descended, but that of grana and lamella increased as a shaded result. Thesize of chloroplast and starch grain of shading leaves minished.

Effect of Potassium on Ultrastructure of Maize Stalk Pith and Young Root and Their Relation to Stalk Rot Resistance

To study the mechanism of potassium （K） application on improvement of maize resistance to stalk rot at cellular level, scanning electron microscope and transmission electron microscope were used to observe the effect of K on the ultrastructure of maize stalk pith tissue and young root tip cell influenced by K and pathogen. In K deficient treatment, parenchyma cells of stalk pith had abnormal structure, and the cell wall between upper and lower adjacent cell was damaged, resulting in the loss of connections between vascular cells and insufficient supporting capacity. However, an improved K nutrition helped to keep a quite tight arrangement of root cell with thick cell wall, and prevent the invasion of pathogen effectively. Moreover, K treated root cell had abundant golgi apparatus, which could excrete large amount of secretions to degrade mycelium. Papillary and highly electronic intensity dot were accumulated at the invading point to prevent the deveJopment of the mycelium. Improved K nutrition could increase the resistant ability of maize plant to stalk rot, through keeping cell structure stability, preventing the expansion of intracellular space to reduce the chances of pathogen invasions, and through reinforcing cell wall and formation of intercellular and intracellular material to restrict further development of pathogen in host cell.

Effects of different LEDs light spectrum on the growth,leaf anatomy,and chloroplast ultrastructure of potato plantlets in vitro and minituber production after transplanting in the greenhouse

Light spectrum plays an important role in regulating the growth and development of in vitro cultured potato(Solanum tuberosum L.) plantlets. The status of potato plantlets at the end of in vitro stage influences the minituber production after transplanting. With 100 μmol m^-2s^-1 total photosynthetic photon flux density(PPFD), a light spectrum study of 100% red light emitting diodes(LEDs) light spectrum(RR), 100% blue LEDs light spectrum(BB), 65% red+35% blue LEDs light spectrum(RB), and 45% red+35% blue+20% green LEDs light spectrum(RBG) providing illumination at the in vitro cultured stage of potato plantlets for 4 weeks using fluorescent lamp as control(CK) was performed to investigate the effects of LEDs light spectrum on the growth, leaf anatomy, and chloroplast ultrastructure of potato plantlets in vitro as well as the minituber yield after 2 months transplanting in the greenhouse. Compared to CK, RB and RBG promoted the growth of potato plantlets in vitro with increased stem diameter, plantlet fresh weight, plantlet dry weight, and health index. Furthermore, BB induced the greatest stem diameter as well as the highest health index in potato plantlets in vitro. Root activity, soluble protein, and free amino acid were also significantly enhanced by BB, whereas carbohydrates were improved by RR. In addition, thickness of leaf, palisade parenchyma and spongy parenchyma was significantly increased by BB and RBG. Chloroplasts under BB and RBG showed well-developed grana thylakoid and stroma thylakoid. Unexpectedly, distinct upper epidermis with greatest thickness was induced and palisade parenchyma and spongy parenchyma were arranged neatly in RR. After transplanting in the greenhouse for 2 months, potato plantlets in vitro from BB, RB, and RBG produced high percentage of large size tuber. BB improved fresh and dry weights of the biggest tuber but decreased tuber number per plantlet. In addition, RBG increased tuber number as well as tuber fresh and dry weight slightly. Our results suggested monochromatic blue LEDs as well as combined red, blue or/and green LEDs light spectrum were superior to fluorescent lamp spectrum in micro-propagation of potato plantlets. Therefore, the application of RBG was suitable;BB and RB could be used as alternatives.

Scleral ultrastructure and biomechanical changes in rabbits after negative lens application

AIM： To address the microstructure and biomechanical changes of the sclera of rabbits after negative lens application by spectacle frame apparatus. METHODS： Five New Zealand rabbits of seven weeks post-natal were treated with -8 D lens monocularly over the course of two weeks. Refractive errors and axial length （AXI.） were measured at the 1st, 7th and 14th days of the induction period. Ultrastructure of sclera was determined with electron microscopy. Biomechanical properties were tested by an Instron 5565 universal testing machine. RESULTS： l.ens-induced （1.1） eyes elongated more rapidly compared with fellow eyes with AXI. values of 15.56±0.14 and 15.21±0.14 mm （P〈0.01）. Fibril diameter was significantly smaller in the I.I eyes compared with control ones in the inner, middle, and outer layers （inner layer, 63.533 vs 76.467 nm; middle layer, 92.647 vs 123.984 nm; outer layer, 86.999 vs 134.257 nm, P〈0.01, respectively）. In comparison with control eyes, macrophage-like cells that engulfed fibroblasts, dilated endoplasmic reticulum, and vacuoles in fibroblasts were observed in the inner and middle stroma in the/U eyes. Ultimate stress and Young＇s modulus were lower in the I.I eyes compared with those in the control eyes. CONCLUSION： Negative lens application alters eye growth, and results in axial elongation with changes in scleral ultrastructural and mechanical properties.

Ultrastructure of the sensilla on antennae and mouthparts of larval and adult Plutella xylostella (Lepidoptera:Plutellidae)

Plutella xylostella （L.） （Plutellidae） is an important agricultural pest throughout the world. In this study, the morphology of antennal and mouthpart sensilla in the larvae and adults of P. xylostella （L.） was observed by using a scanning electron microscope. The larval antennae possess six sensilla basiconica, two sensilla chaetica and one sensillum styloconicum. Larval mouthparts possess six types of sensilla： sensilla chaetica, sensilla digitiformia, sensilla epipharyngeal, sensilla basiconica, sensilla styloconica and sensilla placodea. In the adult, seven types of sensilla are found on the antennae in males and six types of sensilla （sensilla basiconica absent） occur in females. Sexual dimorphism is also found in the number and size of these sensilla on the antennae of adults. We describe for the first time the five types of sensilla on the mouthparts of the adult of P. xylostella. This study provides useful information for further research into the function of these sensilla, and better understanding the behavioral mechanisms involved in pest control.

Taxonomy and ultrastructure of five species of Tetraselmis (Prasinophyceae) isolated from China seas

Comparative ultrastructural investigations on twenty-three isolates of Tetraselmis from the South China Sea, East China Sea and Huanghai Sea, have revealed that these isolates belong to:(1)Tetraselmis chui Butcher; (2)T. cordiformis (H. J. Carter) Stein; (3)T. helgolandica Kylin; (4)T. suecica (Kylin) Butcher; (5)T. guangdongensis sp. nov. Except T. helgolandica, the others are new records in China. T. guangdongensis sp. nov. is a new species. Its external and anatomical features closely resemble those of T. impellucida McLachlan et Parke. As in that species the pyrenoid is penetrated from many directions by cytoplasmic channels delimited by a double membrane. The protoplast withdraws from the apical portion of the theca, that portion which overlaid the trough inverts and pops out in the form of a teat. But unlike that species a starch sheath is present; pyrenoid matrix is surrounded by thylakoids which intervenes between the matrix and the starch sheath surrounding pyrenoid; the theca is stratified.

Cell Ultrastructure of Kiwifruit (Actinidia chinensis) Shoot Tips During Cryopreservation

The changes in the cell ultrastructure of in vitro cultured shoot tips from dwarf genotype of kiwifruit （Actinidia chinensis Ganmi 5） during cryopreservation were investigated. Shoot tips were preserved in liquid nitrogen using vitrification, and the cell ultrastructure was examined using transmission electron microscopy （TEM）. The regular ultrastructure of the cell wall, cell membrane and nucleus of shoot tips could be damaged during the freezing and thawing associated with preservation using liquid nitrogen. The cell plasmolysis was increased and freezing tolerance was improved after precultufing and dehydrating in a preservation and vitrification solution （PVS2） （30% glycerol （Gly）＋ 15% ethylene glycol （EG）＋ 15% dimethylsulfoxide （DMSO） ＋ 0.4 mol L^-1 sucrose）. The structure of some cells with low degree of injury and reversible damage was similar to that of the control and they could undergo normal cell division and differentiation. Besides, they could recover automatically and regenerate after their reculture.

Ultrastructure Observation of Cysticercus Pisiformis and Its Function Analysis

[ Objective] The aim of the study was to observe the ultrastructure of Cysticercus pisiformis under different state. [ Method] The Cysti- cercus pisiformis were obtained in the procedure of slaughter inspection form meat-packing plant. The Cysticercus pisiformis was taken out by cut- ting cyst directly, or cultured with physiologic saline containing 10% bile until evagination. Then it was observed and recorded by scanning electron microscopy. [ Result] Observed from top of scolex of Cysticercus pisiformis locate in cyst, the rostellum looked like an umbrella. The cutaneous muscular columns with hamula covered the front end of scolex. Viewed from the side, a row of antlers-liking hamula adhered to the scolex. Four suckers were looked like more width and body segments less time, in which there were lots of wrinkles in their surface respectively. The Cysticercus pisiformis cultured was divided obviously into scolex, neck, body segment and zone of cyst. When cutaneous muscular column of rostellum con- tracted, the antler-liking hamula extended to periphery that the ring-liking contraction appeared in neck and the lengthways contraction in body seg- ments. [ Conclusion] The Cysticercus pisiformis under different state has distinctly dissimilar ultrastructure.

Aquaporin 4 expression and ultrastructure of the blood-brain barrier following cerebral contusion injury

This study aimed to investigate aquaporin 4 expression and the ultrastructure of the blood-brain barrier at 2-72 hours following cerebral contusion injury, and correlate these changes to the formation of brain edema. Results revealed that at 2 hours after cerebral contusion and laceration injury, aquaporin 4 expression significantly increased, brain water content and blood-brain barrier permeability increased, and the number of pinocytotic vesicles in cerebral microvascular endothelia cells increased. In addition, the mitochondrial accumulation was observed. As contusion and laceration injury became aggravated, aquaporin 4 expression continued to increase, brain water content and blood-brain barrier permeability gradually increased, brain capillary endothelial cells and astrocytes swelled, and capillary basement membrane injury gradually increased. The above changes were most apparent at 12 hours after injury, after which they gradually attenuated. Aquaporin 4 expression positively correlated with brain water content and the blood-brain barrier index. Our experimental findings indicate that increasing aquaporin 4 expression and blood-brain barrier permeability after cerebral contusion and laceration injury in humans is involved in the formation of brain edema.

Calcium Forms, Subcelluar Distribution and Ultrastructure of Pulp Cells as Influenced by Calcium Deficiency in Apple (Malus pumila) Fruits

Calcium in Red Fuji and Starkrimson apples during storage were fractionated by sequentextracting. Localization and distribution of calcium and influence of calcium nutritionon cell ultrastructure were observed by transmission electron microscopy combined within situ precipitation of calcium with an improved method of potassium pyroantimonatetechnique. Results indicated that spraying calcium solution on surface of young fruitsincreased contents of calcium in all forms. During storage, contents of soluble calciumand pectic calcium declined and those in calcium phosphate, calcium oxalate and calciumsilicate increased. Calcium contents of Red Fuji in all forms were higher than those ofStarkrimson, indicating that calcium accumulating capability of Red Fuji fruits precededthat of Starkrimson. Under transmission electron microscopy, calcium antimonite precipitates(CaAP) was mainly distributed in cell wall, tonoplast, nuclear membrane and nucleoplasm,much more CaAP deposited in vacuole. Calcium deficiency during storage leads to decreaseof CaAP in locations mentioned above, disappearance of compartmentation, and entrance ofCaAP to cytoplasm. Transformation from soluble calcium and pectic calcium to calciumphosphate, oxalate and damages of biomembranes structuraly and functionally resultedfrom calcium deficiency during storage were the crucial causation of physiologicaldisorder.

Influence of compound aescine gel on ultrastructure of vein infused mannitol and its mechanism

Purpose: When hypertonic solution 20% mannitol solution was injected into vein, inflammatory mediators and Mitogen-activated protein kinases activated by mannitol can directly induce the fading of vascular endothelial cell, which leads to phlebitis. The study aims to observe the influences of reparil-gel N coated at the proximal parts of the puncture point and basing on this along with infusing heated mannitol to veins to the injure and ultrastructure of veins which were infused the 20% mannitol solution by indwelling needle in vein. Methodology: There are 15 adult New Zealand rabbits. We randomly divided 24 ear veins of 12 adult New Zealand rabbits into Control group, Gelatum group, Gel heated group and injected 20% mannitol solution by vein detained needle in three groups. In Gelatum group, we coated the proximal end of the puncture point with a thin layer of compound aescine gel. Based on Gelatum group, we heated 20% mannitol solution to 35oC-37oC in Gel heated group. Then we observed the intravenous parts and took the veins of each group out to observe their morphology and ultrastructural after the second day of transfusion. 6 ear veins of the rest 3 rabbits as Health group weren’t given any interventions, the veins were taken out to observe their morphology. Results: Comparison between Gelatum group and Gelatum heated group on vascular dilatation, Infiltration of inflammatory cell and Formation of thrombus had no significance, P> 0.05, while the case was different for the comparison on injury of vascular wall, perivascular edema and perivascular hemorrhage, P< 0.05). The statistical significance exists between control group and Gelatum group and Gel heated group, P< 0.05. It was observed under the electron microscope that, in control group, the membrane of endothelial cell peeled off and the mitochondria swelled and vacuolized. In Gelatum group, the membrane of endothelial cell was defective, the parts of the mitochondria were fuzzy, intercellular substance was slightly edematous. In Gel heated group, the mitochondria were clear and intercellular substance slightly swelled. It could be found that the function of phagocyte was complete. Conclusions: Compound aescine gel can prevent phlebitis or reduce the incidence of phlebitis. The combined intervention of coating with a thin layer of compound aescine gel and heating mannitol solution can produce better effect.