Electron-Microscopic Studies in Guinea Pig with Tympanosclerosis / 听力学及言语疾病杂志

Objective To study the middle ear mucosa of guinea pigs with tympanosclerosis by electron microscope and try to obtain some insights into the feature and pathogenesis of tympanosclerosis.Methods Eight healthy variegated guinea pigs were devided into 2 groups.Six(8 ears) guinea pigs were subjected to inoculation of 1×10~8/L of staphylococcus aureus solution 100 μl into the middle ear cavities under the microscope.All the guinea pigs were observed for more than 6 months with no farther treatment.For electron-microscopic studies,the mucosa tissues were taken from the tympanic mucosa in 6 guinea pigs (8 ears) with tympanosclerosis from various sites,while the middle ear mucosa of two healthy guinea pigs (4 ears) were taken as a control.Results Uhrastructural examination of the normal middle ear mucosa revealed a few collagen fibers,normal morphous of fibrocyte,rough endoplasmic reticulum and mitochondria,and there was no lysosome.However,the tympanosclerosis specimens showed that irregular deformation,elongation,and degeneration of fibrocytes and oval nucleus were darkly stained,lots of mitochondria and lysosomes gathered into the cytoplasm around the nuclear and cystic expansion of the endoplasmic reticulum.In the submucosa extracellular matrix,there were a large number of collagen fibers containing lots of amorphous high-density electron-rich body.Conclusion Electron-microscopic studies of the middle ear mucosa of guinea pigs with tympanosclerosis revealed evident proliferation of collagen fibers,and calcifications were seen in the structures such as extracellular matrix vesicle,lysosomes,myelin structures within lipid granules,which mainly in extracellular matrix vesicles.

The effects of hyperbaric oxygen therapy on T cell subpopulation and structure changes of electron microscopy in rats with acute pancreatitis / 中国基层医药

Objective To investigate the effect of hyperbaric oxygen therapy on T-lymphocyte subpopulations in rats with acute pancreatitis. Methods 56 rats were randomly divided into three groups the sham group ( n = 8 ),control group( n = 24) and hyperbaric oxygen therapy group( treatment group, n = 24), then the control group and treatment groups were divided into three subgroups of 8 rats each undergoing euthanasia on days 1,3,7 after the acute pancreatitis induction. The CD4+ ,CD8+ subpopulations of T-lymphocytes in peripheral blood were detected respectively at the fist day and each day of the euthanasia. The sham group was used to make sure that the model was successfully induced. After euthanasia the pancrea was examined using electron microscopy. Results In the control group, the CD4+ cells in AP rats was significantly decreased and the ratio of CD4+/CD8+ also decreased. After 7days of HBO therapy,compared with the control group, the CD4+ lymphocytes of peripheral blood in the treatment group markedly increased( P ＜0.01 ). The CD8+ lymphocytes also increased to a certain extent. And the CD4+/CD8+ ratio of peripheral blood was obviously increased(P ＜0. 01 ). Also more severe pathological changes appeared in the untreated group than in the treatment group. Conclusion Hyperbaric oxygen therapy could improve the oxygen supply in acute panereatitis, regulate T cell immune function.

Morphological Change with the Induction of Cisplatin Resistance from the Bladder Cell Lines / 대한비뇨기과학회지

PURPOSE: When Managing metastatic bladder tumors, to overcome the resistance mechanism of cisplatin is a main problem to be solved. The objective is to confirm the changes of general and ultrastructural morph ology with the induction of cisplatin resistance from the bladder cell line. MATERIALS AND METHODS: The samples of this investigation are 2ng/ml-cisplatin resistant human bladder cell lines T24R2 established by SNUH Urology and the drug resistant bladder cell lines T24 was obtained from ATCC, as a control group. We cultured the resistant cell line on the slide and observed it using light microscopy to see the general morphology. For the ultrastructural morphology, we fixed cultured cells, made an epon block, sliced an ultrathin section and observed it using H-71000 EM. RESULTS: Under light microscopy, the cytoplasm of the resistant cell line shows a plumper pattern than that of the parent cell. Under electronmicroscopy, the chromatin of the resistant cell line has a relatively finely dispersed chromatin pattern when compared to the parent cell line, which shows a coarse and aggregated chromatin pattern. Within the cytoplasm, the mitochondrial volume, dilated rough endoplasmic reticulum, polyribosomes and ribosomes are moderately increased in the resistant cell line when compared to the parent cell line. In particular, we found a great amount of double membrane vesicle near the cell surface and pinocytic vesicles on the surface, which are seldom observed within the parent cells. CONCLUSIONS: We concluded that the cisplatin resistant human bladder cell lines (T24R2) underwent a morphological change with the induction of cisplatin resistance, and we hypothesize that the resistant cell's ultrastructure, which shows morphological change, will be involved in the drug resistance mechanism. Regarding this matter, further research will be needed.

Study on the Action Mechanism of Insulin-Like Growth Factors and HBV-DNA Integration During Hepatocarcinogenesis / 中国医师杂志

Objective To observe the expressions of insulin-like growth factors(IGFs) and proliferating cell nuclear antigen(PCNA) in various kinds of liver tissues,we discussd the relationship among IGFs,PCNA and HBV-DNA integration in the process of hepatocarcinogenesis,provide some theoretical basis for the mechanism of hepatocarcinogenesis and clinical diagnosis and therapy of HCC.Methods 32 cases of HCC,32 cases of paracancerous liver tissues(PLT),12 cases of chronic active hepatitis(CAH),10 cases of fetal liver tissues(FLT) and 8 cases of normal adult liver tissues(NALT) were collected.Southern hybridization was used to detect HBV-DNA integration in various kinds of liver tissues.Then a colloid gold immunoelectron microscopic technique was used to make a systematic study on the expression and ultrastructural location of IGFs and PCNA in different liver tissues.Results The HBV-DNA integration was positively existed in 81 3%(26/32) of HCC and PLT,and 83 3%(10/12) in CAH,but the integration in NALT and FLT was no existed.PCNA was expressed by 87 5%(28/32) of HCC and 84 4%(27/32) of PLT,respectively.The postive rates of IGF-Ⅰ and IGF-ⅠR expressed by HCC were 40 6%(13/32) and 56 3%(18/32),those of IGF-Ⅰand IGF-ⅠR expressed by PLT were 50 0%(16/32) and 65 6%(21/32),respectively.The positive rates of IGF-Ⅱ and IGF-Ⅱ receptor(IGF-ⅡR) expressed by HCC were both 78 1%(25/32),and those expressed by PLT were both 81 3%(26/32).In the integration group of HBV-DNA had higher positive rates of IGF-Ⅱ and IGF-ⅡR,which were expressed by the HCC and PLT groups,than that of non-integration group of HBV-DNA(P

Solitary Fibrous Tumor A clinicopathologic review of five cases

We experienced five cases of solitary fibrous tumor; two in the pleura, two in the orbital soft tissue, and one in the lung parenchyma. Three patients were male, and the age of the patients ranged from 38 to 71 years (mean age: 53.6). Grossly, the masses were well circumscribed and had varying sizes from 2.5 to 30.0 cm. The cut surfaces were grayish-yellow firm with focal variegated hemorrhage, necrosis, cystic change, and myxoid area. Microscopically, these were characterized by a haphazard proliferation of spindle cells or polygonal cells separated by variable amounts of hyalinized collagen and showed a prominent vascular channels reminiscent of hemangiopericytoma in foci. Immunoperoxidase stains showed a strong reactivity for CD34, and were weakly positive for vimentin. Electron microscopical examination revealed features of fibroblast; spindle to round tumor cells were arranged in groups and surrounded by collagen. Nucleoli were seldom prominent. The cytoplasm contained many microfilaments and a moderate number of cisternae of rough endoplasmic reticulum.

A Case of Bullous Systemic Lupus Erythematosus / 대한피부과학회지

Vesiculobullous eruptions are a rare cutaneous manifestation of systemic lupus erythematosus (SLE). They are related to intense inflammation within the cutaneous lesions and subsequent blister formation. We report a case of vesiculobullous eruption of SLE in a 23-year-old male. The bullous eruptions were present ori the lip, forehead, both feet and hands. He met ARA(American Rheumatism Association) is criteria for a diagnosis of SLE and the eruptions developed during a systemic attack of the disease. Routine histology of the lesion showed subepidermal blisters with intact epidermis and dense neutrophilic infiltration in the upper dermis. Direct immunofluorescence of lesional skin showed a linear-granular pattern of immune deposits at the dermoepidermal junction. Electronmicroscopy of the lesional skin showing subepidermal bullae are located beneath the lamina densa. Combination therapy of prednisolone and hydrochloroquine showed complete clearing of the lesions within several weeks. We observed a good response without evidence of recurrence.

Immunohistochemical Stain and Electron Microscopic Study on the Pericytes of Fibrovascular Diabetic Preretinal Membrane

Seven fibrovascular diabetic preretinal membranes were examined with lightmicroscophic immunohistochemical stain and electron-microscopy to evaluate the possibility of pericytes to be involved in membrane contraction. Pericytes were positively stained with anti-actin antibody together with some stromal cells thought to be myofibroblasts presumedly. On transmission electron microscopic study, pericytes were highly active with numerous cytoplasmic processes and contained abundant microfilaments considered as actin in their cytoplasm. Pericyte/endothelial cell ratio of vascular channels were increased in some actively proliferative portion of the membranes. Myofibroblasts that contain abundant cytoplasmic microfilaments were also demonstrated in the extravascular stroma of the membranes and were very similar to the pericytes morphologically. Although the evidence that the pericytes are related to the origin of the myofibroblasts could not be demonstrated, this study suggested that the pericytes may play important roles in development and contraction mechanism of fibrovascular diabetic preretinal membranes.

Hepatitis B virus (HBV) infections in turtles

Thirty turtles (15 Clemys mutica and 15 Geoclemys reevesii) which were inoculated with human sera those were positive for hepatitis B surface antigen (HBsAg) and hepatitis B "e" antigen (HBeAg) were found to be infected with hepatitis B virus (HBV). The levels of HBV infection markers, such as HBsAg and antibody to HBsAg (anti-HBsAg), were retinely monitored in the turtles' serum for 46 weeks. Within two weeks of the inoculation, 42% of the turtles tested were positive for HBsAg, and their reciprocal titers as measured by reverse passive hemagglutination (RPHA) and enzyme linked immunoabsorbance assay (ELISA) ranged from 16 to 96. Within 20 weeks, the remaining turtles tested HBsAg positive, as confirmed by ELISA. At 20 weeks, all but one of the turtles exhibited changes in HBV blood marker from HBsAg to anti-HBs; the one exception was positive for both HBsAg and anti-HBs. At the 47th week, 7 animals were killed and their organs were examined for HBV infected cells utilizing an immunofluorescent technique. Numerous fluorescent cells which reacted with human anti-HBs nad anti-HBc were observed in the following organs: pancreas, liver, kidney, and brain. Histopathologically, edematous changes in hepatocytes and minor cellular infiltration attributed to an inflammatory response were noted. Liver and kidney cells from the infected animals were cultured, and HBV antigen positive cells for HBsAg and HBcAg were detected in the cultures. Throughout the experiment, HBsAg was detected in the supernatant by ELISA. Virus particles which were indistinguishable from Dane particles were seen in the cytoplasmic vacuoles of the cultured cells by electron microscopy. Finally, the presence of HBV DNA was established by molecular hybridization techniques in the culture supernatants of kidney cells from the infected turtles.

Intrathecal levobupivacaine downregulates substance P expression in the distal cerebrospinal fluid contacting neuron for formalin-induced inflammatory pain in rats / 中国药理学通报

Aim To investigate the effects of intrathecal levobupivacaine(LB)on substance P(SP)expression in spinal dorsal horn(L4~5)and the distal cerebrospinal fluid contacting neuron(dCSF-CN)in a rat model of formalin induced inflammatory pain.Methods Twelve male Sprague-Dawley rats(290~310 g,Grade SPF)were randomly divided into 2 groups with 6 in each.Rats were anesthetized with sodium pentobarbital(40 mg?kg-1,ip)and then fixed in the stereotaxic instrument.A 3 ?l volume of 30% CB-HRP to trace dCSF-CN was injected into one of the rats'lateral ventricles.The animals kept alive,forty eight hours later artificial cerebrospinal fluid(ACSF)10 ?l and 0.5% LB 10 ?l(diluted with ACSF)were intrathecally injected in group ACSF and group LB respectively.Inflammatory pain was induced by formalin injection into left hind paw.Then nociception of the animals was assessed behaviorally with the formalin test.Two hours after formalin administration,rats were sacrificed with deeply anesthesia and transcardially perfused.L4~5 segments of spinal cord were removed for immunohistochemical SP-labeling.Brainstem segments containing dCSF-CN were removed for SP/CB-HRP dual-labeling with immunohistochemical procedures and colloidal gold immunoelectronmicroscopy.Results LB significantly reduced paw licking time in the formalin test(P 0.05vsbaseline,P

THE ANGIOARCHITECTURE OF HUMAN ANTERIOR AND POSTERIOR CENTRAL GYRI / 解剖学报

The angioarchitecture of both anterior and posterior central gyri of thirty human brains was studied by means of scanning electron microscopy, double injection method and alkaline phosphatase method. The study was divided into two parts. (a) Pial vessels: The branching pattern of pial arteries on the gyrus surface could be classified into 4 types. Type Ⅰ (bilateral branching type) accounted for 44.44% of the total, type Ⅱ (unilateral branching) 17.99%, type Ⅲ (comb-like) 16.93% and type Ⅳ (irregular) 20.63%. Each gyrus was supplied by several main branches, forming various areas separated by poorly vascularized lines. Two types of anastomoses (general and straight anastomoses) were found. (b) Intracerebral vessels: based on the degree of their penetration, intracerebral arteries and veins were divided into 5 groups, respectively. They were short, intermediate and long cortical, subcortical and medullary vessels. Arteries gave off forward, horizontal and recurrent branches. There were more long cortical arteries in the anterior central gyrus and more intermediate arteries in the posterior. The arteries were not accompanied by veins. Though rich blood supply was found in layer Ⅲ-Ⅳ, there was a broad vascular network with large meshes in layer Ⅴ and Ⅵ in the anterior central gyrus. Problems in distinguishing arteries from veins and particular vascular features were discussed.

THE SYNAPTIC RELATIONSHIPS OF VL NUCLEUS AND CRUCIATE CORTEX OF THE CAT / 解剖学报

The distribution of afferent terminals from VL nucleus to cruciate cortex was studied by using nerve fibers degeneration and electron microscopic technigues. Cortical area 4 could be further divided into three subdivisions according to their geographical relations to the eruciate sulcus: an anterior, a posterior and a lateral zone, respectively. It was demonstrated that the degenerating axon terminals were mainly distributed in anterior and lateral zones. Much less terminals were observed in the posterior zone. Totally 112 terminals were observed in the anterior zone. Among them, 78.6％ were seen in layer Ⅲ, 19.6％ in layer Ⅰ, 1.8％ in layer Ⅴ. Of the 112 terminals found in the lateral zone, 62.5％ were distributed in layer Ⅲ, 31.2％ in layer Ⅰ and 6.3％ in layer Ⅴ. On the other hand, only 33 degenerating terminals were found in posterior zone. their distributions were: 51.6％ in layer Ⅰ, 42.4％ in layer Ⅲ and 6.0％ in layer V. Approximately 75％ of the degenerating axon terminals examined in these three zones made synaptie contacts with dendritic spines, and 25％ with dendritic shafts. The vast majority of these synapses could be classified as asymmetrical type synapse.In area 6, most of the degenerating terminals were encountered in layer Ⅰ and Ⅲ. All of them formed asymmetrical type synapses with dendritic spines. No degenerating terminals could be found in area 3a.

DISTRIBUTION OF MICROTUBULES IN SERTOLI CELL OF RAT TESTIS / 解剖学报

The distribution of microtubules (MTs)in the Sertoli cell of rat testis was studied with indirect immunofluorescence and electron microscopy. We have found that MTs are mainly located in the cytoplasm apical to the nucleus and oriented parallel to the long axis of the Sertoli cell. MTs may extend into the stalks and processes of the cell which embrace different germ cells. With the changes of the architecture of the seminiferous epithelium and the shape of Sertoli cells, there are some regular changes in MTs distribution during the seminifeous epithelium cycle. In stage Ⅰ-Ⅴ, many MTs aggregate around the elongated spermatids and are parallel to their long axis. During stage Ⅶ maturing spermatids are suspended into the lumen by the Sertoli cell processes containing numerous MTs. Some of the MTs conform to the contour of the hook-shaped spermatids heads. After spermiation (stage Ⅶ-Ⅸ), MTs retract from the lumen with the Sertoli cell processes and gather around the spermatids which just start their elongation. These results indicate that the distribution of the MTs in Sertoli cell has close relations with the architecture of the seminiferous epithelium, and the changes of the Sertoli cell shape and the movement of the spermatids.