Objective To investigate the effects of electroacupuncture (EA) at Quchi (LI11) and Zusanli (ST36) acupoints on the ultrastructural structure of cortical neurons in peripheral area and the protein expression of caspase- 3, Bcl- 2, Bax in rats with cerebral ischemia- reperfusion injury. Methods 36 male adult Sprague-Dawley rats were randomly divided into sham operation group, model group, and electroacupuncture group, with 12 rats in each group. The model group and electroacupuncture group were performed with left middle cerebral artery occlusion (MCAO) according to the modified Longa' methods. The electroacupuncture group received electroacupuncture at Quchi (LI11) and Zusanli (ST36) on the paralyzed limb, for 30 minute. The neurobehavioral scores were recorded before and after treatment. The ultrastructural structure of cortical neurons was observed with transmission electron microscope (TEM). The protein expression of caspase- 3, Bcl-2 and Bax were detected by Western blotting technique. Results The neurobehavioral score was lower in the electroacupuncture group than in the control group (P<0.05). Compared with the model group, the chromatin of neurons was even relatively, and the number of mitochondria increased. The expression of Bcl-2 was higher and the expression of caspase-3 and Bax was lower in the electroacupuncture group than in the model group (P<0.05). Conclusion Electroacupuncture at Quchi (LI11) and Zusanli (ST36) acupoints can inhibit the neurons apoptosis in peripheral area through mitochondria-caspase-3 pathway.

BACKGROUND:Mineralized nodules are the mature marker of osteoblast differentiation, and the observation methods mainly use alizarin red staining.
OBJECTIVE:To compare the observation results of mineralized nodules by three methods, and to explore their characteristics and advantages, as wel as further application in the research of bone disease.
METHODS:The rat osteoblast-like cellline UMR-106 were cultured in the fresh medium that was changed every day, for 14 days. Alizarin red staining-light microscope, tetracycline fluorescence labeling-laser confocal scanning microscopy, and scanning electron microscopy were used to observe mineralized nodules. The calcium content of mineralized nodules was quantified using scanning electron microscopy and energy dispersive spectroscopy. In addition, tumor necrosis factor alpha that could inhibit the proliferation and differentiation of osteoblasts was used as the control.
RESULTS AND CONCLUSION:The three morphology methods could be used to observe the mineralized nodules of normal osteoblasts. As for tumor necrosis factor alpha, no mineralized nodules of osteoblasts were observed by alizarin red staining-light microscopy;smal mineralized nodules were observed by tetracycline staining-laser scanning confocal microscopy and scanning electron microscopy, suggesting tetracycline staining and scanning electron microscopy were more sensitive in the observation. Scanning electron microscopy could be used to observe the submicroscopic structures of mineralized nodules in the osteoblasts, and the formation of mineralized nodules, including the calcium secretion. Additional y, scanning electron microscopy combined with energy dispersive spectroscopy analysis can successful y quantify and position the mineralized nodules, indicating a potential application in the research of bone diseases.

BACKGROUND:Tougu Xiaotong capsule is the clinical prescription for the treatment of osteoarthritis, however, its mechanism has not been fuly elucidated. Urokinase type plasminogen activator system which participated in the degradation of the extracelular matrix of articular cartilage and hyperplasia of joint synovium plays an important role in the pathological process of osteoarthritis. OBJECTIVE: To determine the effect ofTougu Xiaotong capsule on urokinase-type plasminogen activator system in knee cartilage tissues of knee osteoarthritis rats. METHODS: Of 144 Sprague-Dawley rats, 120 rats were randomly made into models of knee osteoarthritisvia intra-articular injection of papain, and randomly assigned to model group,Zhuanggu Guanjie Wan group [1.2 g/(kg?d)], low-doseTougu Xiaotong capsule group [0.092 g/(kg?d)], moderate-doseTougu Xiaotong capsule group [0.184 g/(kg?d)] and high-doseTougu Xiaotong capsule group [0.368 g/(kg?d)]. Each group contained 24 rats. Every 2 weeks was considered as a course, with a 2-day interval, totaly 4 courses. The remaining 24 normal rats were included in the blank group. After every two courses, a batch of experimental animals was sacrificed. The pathological changes were observed folowing staining with hematoxylin and eosin. The positive cels of urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor and plasminogen activator inhibitor were measured by immunohistochemistry. The protein levels of urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor and plasminogen activator inhibitor were measured by western blot assay. RESULTS AND CONCLUSION:Mankin’s score was significantly lower in theTougu Xiaotong capsule group and Zhuanggu Guanjie Wan group compared with the model group (P < 0.01), in a time-dependent manner. Immunohistochemical staining indicated that the positive cels of urokinase-type plasminogen activator and urokinase-type plasminogen activator receptor were significantly decreased, but plasminogen activator inhibitor was significantly increased in theTougu Xiaotong capsule group andZhuanggu Guanjie Wangroup in a time-dependent manner. Western blot assay results had an identical trend to immunohistochemistry. These indicated thatTougu Xiaotong capsule showed preventive and therapeutic effects on osteoarthritis by regulating urokinase-type plasminogen activator system.

BACKGROUND:Tougu Xiaotong capsule is the clinical prescription for the treatment of osteoarthritis in Fujian University of Traditional Chinese Medicine, and the previous studies mainly focus on effect to cartilage.
OBJECTIVE:To observe the effect of Tougu Xiaotong capsule on the proliferation and differentiation of osteoblasts as wel as the expressions of bone remodeling correlated factors.
METHODS:Rat osteoblast-like cel line ROS17/2.8 cel s were incubated with Tougu Xiaotong capsule. The ROS17/2.8 cel s were divided into blank control group and Tougu Xiaotong capsule groups with different
concentrations. The cel proliferation was determined by methylthiazolyldiphenyl-tetrazolium bromide assay. Osteoblast differentiation biomarkers alkaline phosphatase activity, osteocalcin and bone mineralized nodules were measured with colorimetry, enzyme-linked immunosorbent assay and alizarin red staining, respectively. The real-time PCR and enzyme-linked immunosorbent assay were used to detect the expressions of bone remodeling factors osteoprotegerin/receptor activator of nuclear factorκB ligand.
RESULTS AND CONCLUSION:Compared with the control group, the Tougu Xiaotong capsule with the
concentration of 0.25-2 g/L could significantly promote the ROS17/2.8 cel proliferation (P<0.05), up-regulate the alkaline phosphatase activity, osteocalcin expression level and mineralized nodules area, and increase the
percentage of bone remodeling factors osteoprotegerin/receptor activator of nuclear factorκB ligand (P<0.05). The mechanism of Tougu Xiaotong capsule protecting osteoarthritis may partly result from the regulation of
proliferation and differentiation of osteoblasts and bone remodeling.

Microscopic and histochemical methods were used to investigate flavonoids localization in the leaf and the stem of the Sarcandra glabra. The results indicated that flavonoids distributed mainly in epidermis, collenchyma, vascular bundles, secretory cells and palisade tissue of leaf. In the stem, they distributed mainly in epidermis, collenchyma, phloem and secretory cells. Histochemical localization of flavonoids using 5% solution of NaOH is convenient, rapid and reliable. The content of flavonoids in the leaf was higher those than in the stem. For sustainable utilization of the resources we suggested that only the leaves could be harvested.
Flavonoids ; metabolism ; Magnoliopsida ; metabolism ; Microscopy ; Plant Leaves ; metabolism ; Plant Stems ; metabolism

OBJECTIVETo investigate the anti-apoptosis effects of Rubus alceaefolius total alkaloids in rats with hepatic injury.

METHODThe hepatic injury model of rat was induced by intraperitoneal injection with CCl4. Sixty SD rats were randomly divided into the normal group, the model group, the R. alceaefolius total alkaloids intervened group, and the bifendate intervened group. The expressions of the levels of liver cell apoptosis were determined by TUNEL. Ultrastructures of the liver cells were observed with transmission electron microscope.

RESULTCompared with the model group, the degree of hepatic injury and the positive expressions of apoptosis in liver tissues in the R. alceaefolius total alkaloids intervened groups and the bifendate intervened group were significantly lower.

CONCLUSIONR. alceaefolius total alkaloids could reduce the pathological changes and degree of hepatic injury in rats, which may be partially through inhibiting the expressions of apoptosis in liver tissue.

Alkaloids ; pharmacology ; therapeutic use ; Animals ; Apoptosis ; drug effects ; Disease Models, Animal ; Female ; Hepatitis, Animal ; drug therapy ; metabolism ; pathology ; In Situ Nick-End Labeling ; Liver ; drug effects ; metabolism ; pathology ; ultrastructure ; Male ; Rats ; Rats, Sprague-Dawley ; Rosales ; chemistry

Objective To study the effect of Jiedu-xiaozheng decoction on theUltrastructure changes and quantitative analysis of SGC-7901 in vitro and its mechanisms. Methods The compound prescription Jiedu-xiaozheng decoction was used to prepare serum containing drug. Serologic pharmacology methods was applied. The decoction was used to co-culture the gastric cells for24 h and 72 h, and the ultrastructure of tumor cells was observed by the transmission electron microscope. Then the parameters of the nucleus and mitochrondria were determined in SGC-7901 cells, including volume density(Vv) ,form factor PE( PE), average volume(V) ,length(L) average surrace area(S),contrasting to cytoplasma. The SIS powervision was applied to analyze their difference. Results Jiedu-xiaozheng decotion can result in reduced microvilli, deflated volume, decreased mitochondrion volume and reduced ridge. The volume density(Vv) ,form factor PE(PE) ,average volume(v) ,length (L) average surrace area(S) reduced in the 24 h group clearly in Jiedu-xiaozheng decoction group compared to control group. Conclusion Jiedu-xiaozheng decoction can significantly induce SGC-7901 cells apoptosis, exhibiting certain early ultrastructure changes such as marginated heterochromatin and mitochondrion pyknosis. The quantity analysis by electronic imaging system may objectively reveal the ultrastructural changes of cell nucleus and mitochondria.

OBJECTIVETo observe Trichomonas vaginalis (T. vaginalis) adhering to and phagocytizing male genitourinary epithelial cells in order to study the pathogenetic mechanism of male trichomoniasis.

METHODSCultured T. vaginalis bodies were incubated with male genitourinary epithelial cells, and then the ultrastructure was observed with transmission electron microscopy.

RESULTST. vaginalis adhered to epithelial cells like amoeba, and formed pseudopodium or surface invagination surrounding or nibbling other parts of the epithelial cytoplasm.

CONCLUSIONST. vaginalis has the speciality of adhering to and phagocytizing to male genitourinary epithelial cells. Genitourinary epithelial cells may be injured directly by the phagocytosis of T. vaginalis. Attention has to be paid to the correlation.

Animals ; Bacterial Adhesion ; Epithelial Cells ; parasitology ; Genitalia, Male ; parasitology ; Humans ; Male ; Microscopy, Electron ; Trichomonas vaginalis ; ultrastructure

Objective To study the ultrastructure of neural stem cell neurosphere cultured in vitro. Methods Neural stem cell neurospheres from the rat brain were observed under transmission electron microscope or with stain of lanthanum nitrate, ruthenium red and tannic acid. Results The conjunction between the cells within neurosphere was loose and no tight junction was observed. Neural stem cells were proliferating lively. Some neural stem cells differentiated into cells with process and structure of axon, even showed the structures similar to myelin.Conclusion The ultrastructure of neural stem cell neurosphere cultured in vitro was revealed.

Objective To investigate the ultrastructural changes of retinal pigment epithelium (RPE) and its permeability in spontaneously hypertensive rats (SHR) and explore the relation between these changes and hypertensive retinopathy. Methods The ultrastructure of RPE cells in the SHR aged five、six、seven months was observed with transmission electronmicroscope and compared to its normotensive control strain(WKY) with the same age.Then, lanthanum tracer procedures were carried out to investigate pathological changes of the blood retinal barrier. Results ①In SHR the main pathological changes involved swelling of mitochondria, enlargement of endoplasmic reticula,decrease of RPE cell infolding, and sparseness of microvilli. These degenerations were more serious in older rats with higher blood pressure.②The breakdown of outer blood retinal barrier with permeation of lanthanum tracers were evident in SHR aged six or seven month, however, in WKY and five month SHR the traces were preven ted from passing by tight junctions. Conclusion The degeneration of RPE owing to ischemia and anoxia arises in early period of hypertensive retinopathy. The pathological changes of ultrastructure and permeability might interact with the damage of visual cells and play a main role in the hypertensive retinopathy.