Congenital myopathies: The current status

Within the history of neuromuscular diseases (NMD), congenital myopathies (CM) represent a relatively new category introduced in the mid-nineteen hundreds upon advent and subsequent application of enzyme histochemistry and electron microscopy by establishing the three major CM, central core disease, nemaline myopathy, and centronuclear myopathy which later pluralized each when the molecular era began at the end of last century. Quickly, during the following 5 decades, many new CM entities were described, based on muscle biopsies and their CM-characteristic myopathology, the former a prerequisite to recognizing an individual CM, the latter of the nosological hallmark of the individual CM. When the molecular era ushered in immunohistochemistry the spectrum and nosography of CM altered in that some CM became allelic to other cohorts of NMD, e.g., congenital muscular dystrophies, other muscular dystrophies, distal myopathies based on different or identical mutations in the same gene. The nosological spectrum of a defective gene also enlarged by recognizing several entities with mutations in the same gene, and same or similar nosological conditions originated from mutations in different genes. Lately, however, CM were reported which lacked any individual myopathological hallmarks, but were clearly based on molecular defects, a fair number of them being newly identified ones. Few CM still remain without any molecular clarification. This nosographic development rendered the original definition of such new CM questionable and brought uncertainty to their classification and nomenclature.

Basic requirements to establish a neuromuscular laboratory

Histopathological analysis of muscle biopsy is a prerequisite in the evaluation of neuromuscular disorders, particularly inflammatory myopathies, metabolic myopathies, congenital myopathies, muscular dystrophies and differentiating myopathies and neurogenic disorders with overlapping clinically features. It not only provides useful information that helps in the diagnosis but also treatment and management. Fundamental skills and basic knowledge regarding handling, processing and analyzing a muscle biopsy are required in any specialized or a general pathology lab supporting neuromuscular clinical services. Care during transport of the muscle biopsy, sample receipt in the laboratory and grossing is very important. Standard operating procedure should be followed for the preanalytical steps (freezing and cryomicrotomy), routine and special staining (enzyme and non enzymatic) and immunohistochemistry. A well organized neuromuscular laboratory with good quality management system is necessary for the practice of myopathology. This article gives an overview of establishing such a laboratory.

Methods and advances in the research of lymphatics in the head and neck / 中华耳鼻咽喉头颈外科杂志

The lymphatic system plays an important role in human health and disease. In addition to a role in the immune response, the lymphatics can also serve as a pathway for cancer metastasis.Head and neck squamous cell carcinoma (HNSCC) is one of the most common cancers.In the case where the primary focus has been controlled, cervical lymph node metastasis is one of the most important factors affecting the prognosis of head and neck cancer.Thus, understanding the anatomy of the lymphatic system is of paramount importance in predicting cancer metastasis and to perform proper lymph node dissection in cancer patients.The objective of this review is to summarize current imaging approaches that facilitate both basic science and clinical investigations of lymphatic vasculature.Mutation analysis of the laryngeal lymphatic system may provide a theoretical basis for the diagnosis and treatment of HNSCC.

The Morphological Study of Muller Cells with Enzyme Histochemical Staining in Retina of Induced Diabetic Rats

PURPOSE: To evaluate the morphological changes in Muller cells of an induced diabetic rat model with carbonic anhydrase histochemical staining. METHODS: Retinae of three normal rats and four streptozotocin-induced diabetic rats were used. The morphological changes in the Muller cells of these retinae were observed using enzyme histochemical staining. RESULTS: The numbers of positive staining Muller cells in diabetic rats retinae were significantly lower than those of the normal rats. In addition, the shape of the Muller cell bodies in the streptozotocin-induced diabetic rat model's retina changed from polygonal to abnormally flat. Furthermore, the staining of the Muller cells' segment in the outer nuclear layer of the diabetic rat's retinae were weaker, and some Muller cell segments were not stained at all. CONCLUSIONS: The numbers of Muller cells in diabetic rats' retinae were significantly lower than those of the normal rats. In addition, the features of Muller cell bodies of the diabetic rats were changed morphologically.

Histological, Enzyme Histochemical, and Electron Microscopic studies of the Rat Kidney Following Administration of Folic Acid

To investigate the morphologic evidence of acute renal failure by folate, histological, histochemical (PAS), enzyme histochemical (Na-K-ATPase, G6PD, and ALP), and ultrastructural studies were performed. The results are as follows: l) Oliguria was most severe 3 hours after folate and the urine volume was 24.8% that of the control group. 2) Histologically, dilatation of tubules, degeneration and focal necrosis of the cortical tubules, and PAS(+) droplets in the tubular lumen were noted. And also frequent mitoses, mild interstital connective tissue proliferation, and neutrophilic infiltrates were observed in the late stage. 3) On enzyme histochemical examination, the activities of Na-K-ATPase and ALP were decreased, but G6PD activity was increased in comparison with the control group. 4) The ultrastructural studies revealed cytoplasmic vacuoles, apical cytoplasmic blebbing, dense bodies, mildly swollen mitochondria, dilated endoplasmic reticulum, loss bf brush border of the proximal tubules, and loss of microvilli of the thin limb of Henle's loop. Later, marked attenuation or loss of infoldings of basal plasma membrane of the cortical tubules was recognized. According to above results, the cause of acute renal failure by late is thought to be the injuries of tubular epithelial cells including sodium pump secondary to tubular obstruction.

Enzyme-histochemical observation on influence of suspension concentrate of niclosamide in Oncomelania hupensis snails / 中国血吸虫病防治杂志

Objective To observe the influence of suspension concentrate of niclosamide on the enzyme activity of Oncomelania hupensis in order to explore its molluscicidal mechanism. Methods Oncomelania hupensis snails were collected in the habitates of river marshland in Dantu District, Zhenjiang City, Jiangsu Provence and were divided into 2 groups. The snails of the treated group were sprinkled with 25% suspension concentrate of niclosamide. The snails of the control group were sprinkled with distilled water. The soft body tissue of the snail was separated and the sections of snail tissue were made in the Cryostat Microtome. The stain of enzyme-histochemistry showed CCO, LDH, SDH, CHE and NOS had been done, and then the staining block was made by routine method. The staining reaction in the snail tissue and the average gray density were observed with the image analysis system of biomicroscope. Results The enzyme activity of CCO, LDH, SDH, CHE and NOS located in the mouth, muscle fiber, tegumentary membrane, ganglia, liver and pharyngeal cavity of Oncomelania hupensis snails. The enzyme activities of CCO, LDH, SDH, CHE and NOS in the treated group were significantly lower than that in the control group. Conclusions Niclosamide can affect the transmitting of neurohypophysis and obstruct the energy and result in the disorder of the physiological functions in Oncomelania hupensis. It is one of the reasons of Oncomelania hupensis death.

Study on histochemical localization of phenol oxidase in Oncomelania hupensis / 中国血吸虫病防治杂志

Objective To investigate the histological distribution of phenol oxidase (PO) in body of Oncomelania hupensis. Methods Oncomelania hupensis snails collected from the riverside of Wuhu section of the Yangtze River were anatomized and the unimpaired software organs were sliced in frozen condition to get the continued sections, then the sections were incubated in pH 6.8 phosphate buffer containing 25 mmol/L catechol at 37 ℃ and investigated systematically for histological distribution of PO in snail bodies. Results The light microscope showed that there were black grannles of PO (histochemical reaction) in the liver, ctenidium, head-foot, tentacle and mantle collar of snail body. Conclusion PO exists in Oncomelania hupensis and locates in the liver, mantle collar, head-foot, tentacle and ctenidium.

Studies on phenol oxidase in Oncomelania hupensis and its histochemical localization / 中国血吸虫病防治杂志

Objective To understand whether phenol oxidase (PO) existed in the body of Oncomelania hupensis and to observe histological distribution of the enzyme. Methods Enzyme histochemical method: adult female and male snails collected from the riverside of Yangtze River of Wuhu section were anatomized and consequently incubated in pH 6.8 phosphate buffer containing 25 mmol/L catechol at 37 ℃. Then under a stereo microscope, some of the snails were observed and recorded for the existing condition and histological distribution of PO in their bodies. Fluorescent histochemical method: after the process of incubation mentioned above, catechol were exsucted from the group of other snails. Afterward, some phosphate buffer solution (pH 6.8) containing 0.05% sodium penobarbital was added into the group. Then the existing condition and histological distribution of PO were also observed and recorded under a fluorescent microscope. Results The first method indicated that the color of the liver surface turned from French grey to light grey in 5 minutes, to grey in 15 minutes and black in 30 minutes. The second method showed that fluorescien appeared on the surface of the snail liver. Conclusion PO exists in the bodies of male and female snails and locates in their livers.

A MORPHOLOGICAL STUDY OF THE EFFECT OF ACUTE EXPERIMENTAL ISCHEMIA ON THE PYRAMIDAL CELLS OF CANINE CEREBRAL CORTEX / 解剖学报

In the present study, 10 healthy adult dogs were used,8 for the experimental group and 2 for the normal group. We observed the histochemical and ultrastrac- tural changes of the enzymes in the pyramidal cells in the cerebral cortex under the condition of acute ischemia. The result demonstrated that the activity of LDH, AcP and AchE was increased, while the activity of SDH, MAO, ATP ase was decreased. The ultrastructural chnges showed that in the acute ischemia gromp, there were enlargement perinucleus spaces of neurone, brisement, disappearance and vasicularization of the mitochondrial crista. This indicated that acute ischemia had an obvious effect on the histochcmistry of enzymes and the ultra- structure of the neurone in the cerebral cortex.

A STUDY OF THE ENZYME ACTIVITY IN ISCHEMIC AREA OF HEART MUSCLES CAUSED BY THE EXPERIMENTAL CORONARY ARTERY SPASM INDUCED BY THE PITUTRIN INJECTION / 中国法医学杂志

Diagnosis of acute heart ischemia induced by coronary artery spasm(CAS)is not clear yet.We have systematically studied the histopathological,enzyme-histochemical and ultrastructural of the autolytic and the ischemic changes ofthe raf's heart induced by the injection of the pitutrin into the rat's sublingualvein.The PTAH stain demonstrated that some irregular transverse bands hadappeared in the muscular fibers.The adenosine triphosphatase activity in smallarteries was decreased.The author suggests that the results are helpful fordignosis of acute heart ischemia caused by CAS.

HISTOCHEMICAL OBSERVATION ON EXPERIMENTAL RAT'S EARL MYOCARDIAL INFARCTION / 中国法医学杂志

This Paper reports the histochemical changes occcured at the early acute cardial ischemic area in-duced experimentally by ligation of left coronary artery of 50 male Wistar rats. The following histo-chemical changes at the ischemical areas were observed: obvious decrease of glycogen of cardical is-chemical for 1h, of both NADHD and LDH activity of ischemical for 2h as well as CCo,CK and ATP activity of ischemical for 4h. The decrease of CCo,CK and ATP activity propagated to the whole is-chemical area at the time of ischemical for 8h. The activity of NADHD,LDH'CK and ATP can toler-ated the postmortem autolysis influence up to 24h after death.

ENZYME HISTOCHEMICAL STUDY OF EXPERIMENTAL RABBIT INCISED WOUND / 中国法医学杂志

Enzyme-histochemical investigations on experimental incised injury of thirty rabbits fortiming of wound were reported.In peripheral areas of wounds,strong reaction of six kinds ofdehydratase(ACP.AKP,ATP.ANAE.?—GA,?—Gr).were demonstrated 1～2 hrs followingincision while no reaction of four kinds of dehydrogenase(LDH.SDH.NADH.?—GPDH)werefound.The activity of dehydratase are explained by the enhanced metabolism of fibrocytes and theaggregation of acute inflammatory cells.The author suggests that the positive enzyme histochemicalreactions indicate the vital reaction in the first two hours,and PTAH staining is helpful for thediagnosis of micro—exudation of fibrin in the early stage of injury

Experimental Studies on Disuse Atrophy of the Rat Tibialis Anterior Muscle / 대한정형외과학회잡지

The experiment was designed to investigate type-selectivity of the involved muscle fibers and changes in the component ratios of the fiber types following the progression of disuse atrophy in the skeletal muscle. After inducing disuse atrophy of the skeletal muscle by applying cast on the hind limb, we used histological and histochemical stains in the sections of the rat Tibialia anterior muscles. The results were as follows. 1. Even 8 weeks after immobilization of the hind limb, differentiation of muscle fiber types by histolo gical and histochemical staining methods in the Tibialis anterior muscle could be possible. 2. Atrophy of muscle fibers was more pronounced in type IIB and type I fibers than in type IIA fibers. 3. Central migration of sarcolemmal nuclei and ring fiber appeared after 6th and 8th weeks of immobilization respectively, in the H&E and trichrome preparations. Ac-pase or Alk-pase positive fibem were not noted throughout the experimental periods.

Enzyme Histochemical Study for the Estimation of the Lapse of Time in Brain Injury

This experiment was designed for the evaluation of the usefulness of enzyme histochemistry in the determination of the lapse of time in brain wound, and also for the establishment of medicolegal 'biological time table' on brain wound. Brain injury was made by contusion and laceration of meninges and brain itself in rats. The results were as follows; 1) By routine histological technique, estimation of the lapse of time in brain wound could be possible 4 hours after the infliction of wound. 2) The earliest change of enzyme activities was recognizable by the decreased activities of ATPase and succinic dehydrogenase 30 minutes after the injury. These decreased enzyme activities were not recovered up to the 4th day after the brain injury. 3) Increased acid phosphatase activity was noticed 1 hour, and beta-glucuronidase, 2 hours after the injury in a mild degree. Both increased activities were pronounced following the lapse of time in brain wound. 4) No significant change was seen in alkaline phosphatase, monoamine oxidase, non-specific esterase and leucine aminopeptidase activities throughout the experimental period up to the 4th day. So the enzyme histochemistry of these enzymes seemed to be little valuable for the study on the timing of wound in brain injury. In the light of these results it appeared that the enzyme histochemistry, in particular of ATPase, succinic dehydrogenase, and acid phosphatase, for the estimation of timing of brain wound not only shortened the histological "lag period" up to 30 minutes after the injury, but also provided a useful information in determining the biological time table following the brain injury.

AN ENZYME HISTOCHEMICAL INVESTIGATION ON THE CHANGES IN ENTERIC NERVE PLEXUS OF GUINEA PIGS AFTER VAGOTOMY / 解剖学报

0.05). This study suggest that unlike other autonomic ganglia, the enteric nerve ganglia may exhibit a relatively high capacity of autonomic regulation and compensatory adaptation, and also provide some histochemical evidences for the transneuronal degeneration changes.

HISTOLOGICAL AND ENZYME HISTOCHEMICAL STUDIES OF THE RAT THYROID GLAND DURING EXPERIMENTAL GASTRIC ULCER / 解剖学报

Eighty adult male Wistar rats were randomly divided into three groups: 1. In the experimental peptic ulcer group, 35 animals were induced to develop peptic ulcer by injecting 0.05 ml/kg of glacial acetic acid (more than 99%) into the submucosa of the stomach after laparotomy under aseptic conditions. 2.In the saline control group, 35 animals were injected with 0.05 ml/kg of saline into the submucosa of the stomach after laparotomy. 3.In the normal control group, 10 normal animals without any treatment were raised under the same condition as group 1 and group 2. The thyroid glands of three groups were taken at definite time intervals (1-28 days) after the operation. The right thyroids were prepared for cryostat sections after hexane quenching (-60℃) and subjected to the histochemical studies of acid phosphatase (AcP), alkaline phosphatase(AIP), a-glycerophosphate dehydrogenase (a-GPD), succinate dehydrogenase (SDH), glucose-6-phosphate dehydrogenase (G6PD), and nonspecific esterase (NsE). The left thyroids were fixed in Carnoy's fluid, stained with HE and subjected to histological study. The reactions of enzymes of the follicular cells in the group 2 were weaker than those in the group 3 (normal control) during the period of 2-21 days after the operation. The follicular cells became flattened and follicular lumens increased in size. These changes recovered to normal on the 28th day after the operation. In the follicular cells of group 1. In which peptic ulcer developed after injection of glacial acetic acid, the reaction of A1P was weaker than those in the group 3, but stronger than those in the group 2. The reations of a-GPD, SDH, and G6PD were stronger than those in the group 2, and as well as those in group 3. The reaction of AcP was stronger than those in the group 2 and group 3 during the 6-21 days after the operation. The follicular cells in the group 1 became flattened and the follicular lumens increased in size only during the period of 4-10 days after the operation and recovered on the 14th day after the operation. These findings suggested that the thyroid follicular cells of rat involved in the metabolic activities of the repair of gastric ulcer.

HISTOCHEMICAL STUDIES ON ENZYMES CONCERNED WITH PHOSPHATE ESTER, ESTER AND NEUROTRANSMITTER METABOLISM IN THE ENTERIC PLEXUS OF GUINEA PIG / 解剖学报

Enzyme histochemistry of neurons in the enteric nerve plexus of guinea pigs were studied with light microscope semi-quantitatively and microphotometer quantitatively. The results showed that the neurons differ greatly in A1P (alkaline phosphatase), AcP (acid phosphatase), 5'-Nase (5'-Nucleotidase), TPPase (thiamine pyrophosphatase), NsE (non specific esterase) and ChAT (choline acetyltransferase). There were disparities to a certain extent in reactions of MAO (monoamine oxidase), AP (aminopeptidase) and AChE (acetylcholinesterase) among different segments of gastrointestinal between submucous and myenteric plexus, but all neurons were positive for the enzymes stated above. The neurons in each ganglion were relatively similar in the enzyme activities. There were about 50-66% neurons in the enteric nerve plexus showing strong reaction of ChAT, which may be cholinergic neurons. There were significant differences in enzymatic activities, except NsE, between submucous plexus and myenteric plexus statistically. Submucous plexus showed stronger reactions of AcP and AP than those of myenteric plexus, while myenteric plexus showed stronger reactions of A1P, 5'-Nase, TPPase, MAO, ChAT than those of submucous plexus. The ganglia of intramural plexus in stomach were not well developed as those of intestine, especially the submucous plexus of stomach, in which there were only few scattered neurons, and they showed weaker enzyme activities than those of intestine. The enteric neurons in duodenum and proximal colon showed strongest activities for most enzymes among different segments of intestine. The above results indicate that the enteric neurons exist remarkable differences in metabolism and functional states.

OBSERVATIONS ON THE MORPHOLOGICAL CHANGES OF EPIDERMAL LANGERHANS CELLS IN REPEATED SKIN IRRITATION TEST WITH BENZALKONIUM BROMIDE AND DINITROCHLOROBENZENE / 解剖学报

The kinetic changes in morphology and cellular density of epidermal Langerhanscells (LC) in the guinea pig were observed by ATPase cytochemical staining techniqueafter repeated skin application of 5%,3% and 1% of benzalkonium bromide (BB,primary irritant) and 0.05% dinitrochlorobenzene (DNCB,allergen) respectively.We have found that there was a significant difference in the density and morpho-logical change of LC between BB and DNCB application.Treatment with 5% BBcould induce a reversible decline in LC density and changes in cell processes,andwith 0.05% DNCB,the number of LC was decreased and the ATPase activity wasweakened only in the late stage of treatment.The significance of using the kineticchanges in morphology and cellular density of LC as the criteria for the safetyevaluation of weak allergens and weak primary irritants as well as cosmetics wasdiscussed.

HISTOCHEMICAL STUDY OF ENZYME OF AAND B-CELLS OF PANCREATIC ISLETS DURING EXPERIMENTAL PEPTICULCER IN RATS / 解剖学报

Sixty five adult male rats were selected for the experiment. The rats were divided into three groups. Peptic ulcer was induced in the experimental group by means of injection of acetic acid to the submucosa of the stomach. Animals of the control group were injected with normal saline instead. Another control group consisted of normal rats without any treatment. All rats were killed at different time intervals (2-28 d) after the operation. The pancreas tissues were prepared for enzyme histochemical study. The results were observed as follows: Two to four days after the peptic ulcer operation, in the pancreatic A-cells, the activity of alkaline phosphatase (A1P) decreased and 5-nucleotidase (5-Nase) activity increased. A1P, adenosinc triphosphatase(ATPase),succinic and lactic dehydrogenases(SDH, LDH), glucose-6-phosphate and a-glycerophosphate dehydrogenases(G-6-PD,orGPD)acuvities were rasied in varying degrees and returned to normal level after 28 days of the operation. In the pancreatic B-cells, during the early stage of the peptic ulcer, acid phosphatase (AcP) and 5-Nase activities increased and ATPase activity declined. But after four days, AcP, 5-Nase, SDH and LDH activities declined, then began to recover and returned to normal level after twenty one days of the operation. In the normal saline operation control group, the changes of A- and B-cells, two to four days after the operation were the same as in the peptic ulcer group and returned to normal level during the period of six to ten days. The above histochemical changes showed that the pancreatic A- and B-cells played a certain role in the recovery of the experimental peptic ulcer.

A MORPHOLOGICAL STUDY ON TWO TYPES OF ALVEOLAR MACROPHAGES FROM NORMAL RAT LUNG / 解剖学报

Alveolar macrophages (AM) were studied by means of phase contrast microscope, differential interference contrast microscope, SEM and TEM. The content of AcP, LDH, SDH of the AM was measured by MPV3-TSA microspectrophotometerimage analytical instrument. Under phase contrast microscope, two different types of AM can be distinguished, i. e, the light cells and the dark cells. The two types of cells are spherical and flat in shape under differential interference contrast microscope. The observation with SEM and TEM showed that the spherical cells possess more filopodia and more lysosomes in cytoplasm, but the flat cells possess more lamellar podia. Quantitative cytochemistry demonstrated that the content of AcP, LDH, SDH in spherical cells are much more than that of flat cells. The results suggest that there are two types of alveolar macrophages exist in normal rat lung.