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ABSTRACT: To summarize the improvement of various muscle staining techniques and discuss their application in diagnosis of neuromuscular diseases.
Three hundred cases of skeletal muscle biopsy samples were examined by histopathological methods. The flash-freezing techniques were used for the preparation of frozen section, which were stained with HE, Gomori trichromic (GMR), glycogen (PAS), and fat acid (oil red O); the enzyme histochemical staining with myosin adenosine triphosphatase (ATPase), and NADH-TR. Those stained methods had been improved. The immunohistochemical staining with dystrophin; and transmission electronic microscopy were used.
Deepfreeze with heteropentane-liquid nitrogen and flash-freezing techniques could avoid artifacts of ice crystal vacuolation. GMR staining mainly showed the degenerative and necrotic lesions of mitochondria and muscle fibers. Oil red O staining showed the increase of lipid in muscle fibers. PAS staining mainly showed glycogen and glycoprotein. Application of frozen sections in muscular tissue was better than that of paraffin sections. NADH-TR staining and ATPase staining could distinguish the two types of muscle fibers, and show the changes of inner structure of muscle fibers and mitochondria enzymes.
The distribution and characteristic pathological changes of the two types of muscle fibers can be showed clearly by enzyme and non-enzyme histochemical staining techniques of the skeletal muscle. These methods can compliment with each other. Only after understanding the technical principles can we master and apply these methods.
Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences 09/2009; 41(4):459-62.
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Zhonghua bing li xue za zhi Chinese journal of pathology 12/2006; 35(11):697-8.
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ABSTRACT: To compare the advantages and disadvantages of three staining methods of HBV.
Normal Liver tissue and HBV-infected, HCV-infected, or dually infected (HBV and HCV) liver tissues were selected for this study. Formalin-fixed, paraffin-embedded sections (4 microm) were prepared. Each of the liver tissue specimens was detected by three staining methods, including immunohistochemical methods, Shikata's orcein stain and Victoria blue stain,respectively.
In the three methods, all of six HBV-infected cases showed intense staining, and three cases with dual infection (HBV and HCV) were weakly positive. However, both normal and HCV-infected liver tissues showed no staining. HBsAg stained dark brown with Immunohistochemical stain; HBsAg containing ground-glass hepatocytes stained magenta with Shikata's orcein stain; HBsAg stained blue with Victoria blue.
Each of three methods has its own advantages and disadvantages: high specificity and sensitivity, but high cost for immunohistochemical methods; complicated and overelaborate procedure for preparation of solutions, lower specificity and sensitivity, but low cost, for special staining methods.
Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences 07/2006; 38(3):318-20.
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ABSTRACT: To study the clinicopathological features and differential diagnoses of mixed epithelial and stromal tumor of the kidney.
Clinical and pathological characteristics of 4 cases of mixed epithelial and stromal tumor of the kidney were studied.
Three patients were female and one was male. All patients presented with flank pain and hematuria. Radiologic studies revealed cystic and solid masses involving the kidney. Grossly the tumors had a solid and cyst appearance. Microscopically, the tumors were composed of a mixture of stromal and epithelial elements. The epithelial elements were variable in cell types including cuboidal, hobnail and columnar cells. One case showed Müllerian and intestinal epithelial differentiations. Stromal elements essentially consisted of spindle cells, with thick-walled blood vessels and bands of smooth muscle cells as distinctive features of the tumor. Immunohistochemical staining revealed that the epithelial components were positive for AE1/AE3, whereas the stromal components were positive for ER, PR, and SMA. All patients underwent nephrectomy and were well without evidence of recurrence.
Mixed epithelial and stromal tumor of the kidney is a benign neoplasm with distinct histopathological features. It should be distinguished from many other renal neoplasms. Surgical intervention is a preferred therapy.
Zhonghua bing li xue za zhi Chinese journal of pathology 02/2006; 35(1):29-31.
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Fei Pei,
Jie Zheng,
Zi-fen Gao,
Yan-feng Zhong,
Wei-gang Fang,
En-cong Gong,
Wan-zhong Zou,
Sheng-lan Wang,
Dong-xia Gao,
Zhi-gang Xie,
Min Lu,
Xue-ying Shi,
Cong-rong Liu,
Jing-ping Yang,
Yu-ping Wang, Zhi-hui Han,
Xiao-hong Shi,
Wen-bin Dao,
Jiang Gu
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ABSTRACT: Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first manifested in humans in November 2002. The SARS-associated coronavirus (SARS-CoV) has been identified as the causal agent, but the pathology and pathogenesis are still not quite clear.
Post-mortem lung samples from six patients who died from SARS from April to July 2003 were studied by light and electron microscopy, Masson trichromal staining and immunohistochemistry. Evidence of infection with the SARS-CoV was determined by reverse-transcription PCR (RT-PCR) , serological examination and electron microscopy.
Four of six patients had serological and RT-PCR evidence of recent infection of SARS-CoV. Morphologic changes are summarized as follows: (1) Diffuse and bilateral lung consolidation was seen in all patients (6/6) with increasing lung weight. (2) Diffuse alveolar damage was universal (6/6) with hyaline membrane formation (6/6), intra-alveolar edema/hemorrhage (6/6), fibrin deposition (6/6), pneumocyte desquamation (6/6). A marked disruption in the integrity of the alveolar epithelium was confirmed by immunostaining for the epithelial marker AE1/AE3 (6/6). (3) Type II pneumocytes, with mild hyperplasia, atypia, cytomegaly with granular amphophilic cytoplasm and intracytoplasmic lipid accumulation (5/6). (4) Giant cells in the alveoli were seen in five of 6 patients (5/6) , most of which were positive for the epithelial marker AE1/AE3 (5/6), but some cells were positive for the macrophage marker CD68(2/6). (5) A pronounced increase of macrophages were seen in the alveoli and the interstitium of the lung (6/6), which was confirmed by histological study and immunohistochemistry. (6) Haemophagocytosis was present in five of the 6 patients(5/6). (7) Lung fibrosis was seen in five patients(5/6), with alveolar septa and interstitium thickening(5/6), intraalveolar organizing exudates (6/6) and pleura thickening (4/6). Proliferation of collagen was confirmed by Masson trichromal staining, most of which was type III collagen by immunostaining. The formation of distinctive fibroblast/myofibroblast foci was seen in five patients (5/6) by light microscopy and immunochemistry. (8) Squamous metaplasia of bronchial mucosa was seen in five patients(5/6). (9) Thrombi was seen in all patients(6/6). (10) Accompanying infection was present in two patients, one was bacteria, the other was fungus. In addition, electron microscopy revealed viral particles in the cytoplasm of alveolar epithelial cells and endothelial cells corresponding to coronavirus.
Direct injury of SARS-CoV on alveolar epithelium, prominent macrophage infiltration and distinctive fibroblast/myofibroblast proliferation may play major roles in the pathogenesis of SARS.
Zhonghua bing li xue za zhi Chinese journal of pathology 11/2005; 34(10):656-60.
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ABSTRACT: To study the morphologic changes of collagen type III glomerulopathy and to investigate the possible cellular origin for collagen III production.
Light microscopy, immunofluorescent staining, immunohistochemistry (for collagen I, III and IV and alpha-SMA) and electron microscopy studies on 3 renal biopsy cases of collagen type III glomerulopathy were performed.
Two cases presented with nephrotic syndrome, one of which was associated with systemic hypertension. The third case showed renal impairment and renal hypertension. None had any known family history of renal diseases. Light microscopy showed diffuse thickened glomerular basement membrane and expanded mesangium with deposition of weakly PAS-positive homogeneous material not associated with mesangial cell proliferation. Electron microscopy revealed massive collagen fiber deposits in the subendothelial spaces and mesangium. The mesangial cells also contained bundles of microfilaments in the subplasmalemmal regions. Immunohistochemically, the diffuse positivity for type III collagen corresponded to the homogeneous material seen under light microscopy. The staining for type I and IV collagens was negative. Alpha-SMA was expressed in many mesangial cells.
The diagnosis of collagen type III glomerulopathy can be made on the basis of detailed morphologic examination and ancillary investigations. It is possible that activated mesangial cells may be the cellular origin of collagen III.
Zhonghua bing li xue za zhi Chinese journal of pathology 08/2005; 34(7):385-8.
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ABSTRACT: To detect the expression of macrophage inflammatory protein-1alpha (MIP-1alpha), a disintegrin-like and metalloproteinase (ADAM) 8 and 12 and CD68 protein in giant cell lesions of jaw and giant cell tumors of long bone, and to study their effects on the histogenesis of giant cells in such lesions.
MIP-1alpha, ADAM8, ADAM12 and CD68 were detected by immunohistochemistry in 24 paraffin-embedded specimens of central giant cell lesions of jaw and giant cell tumors respectively.
MIP-1alpha positive signal was located in blood vessels and bone. ADAM8, ADAM12 and CD68 positive signals were located in the cell membrane and cytoplasm of all multinucleated giant cells and some round mononuclear cells in the lesions. In addition, some spindle mononuclear stromal cells were positive for ADAM12 in both lesions.
Multinucleated giant cells probably originate from CD68-postive round mononuclear cells, which are recruited from monocyte-macrophage system by chemokines, such as MIP-1alpha, followed by cell fusion mediated by ADAM8 and ADAM12.
Zhonghua bing li xue za zhi Chinese journal of pathology 08/2005; 34(7):393-6.
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ABSTRACT: In order to clarify the exact molecular weight of tumor metastasis suppressor gene-1 (TMSG-1) protein and its cellular localization, a monoclonal antibody against TMSG-1 was prepared, characterized and applied to evaluate the metastatic potential of human tumors.
A dominant epitope-TMSG-1(15)-derived from TMSG-1 was synthesized based on Fmoc method, and the hapten was conjugated to Imject Maleimide activated mcKLH as a carrier protein. The antigen preparation was used to immunize BAL B/C mice. Hybridomas were generated and screened by ELISA for specific monoclonal antibodies, which were further characterized by western blotting and immunohistochemical staining.
One hybridoma cell line secreting anti-TMSG-1 antibody, designated as C8, was eventually established after primary ELISA screening, followed by rapid limited dilution procedure. It was confirmed that C8 was of IgM isotype. Result of competitive inhibition assay showed that the antibody was TMSG-1 specific. Using this antibody, an expected protein band of about 45,000 (relative molecular mass) was detected in the non-metastatic variants PC(3)-2B4 and PG-LH7 cells by Western blotting, but not in the isogenetic metastatic variants of PC3-1E8 and PG-BE1 cells. Immunohistochemistry using C8 showed a positive staining of cell membrane and cytoplasm of 2B4 and LH7 cells, whereas 1E8 and BE1 cells were non-reactive. Immunostaining using C8 of paraffin sections of 52 breast carcinomas and 41 colon cancers demonstrated a strong positivity in non-metastatic tumors, but none to weakly reactive in metastatic tumors (P < 0.05).
C8 monoclonal antibody against the synthetic peptide is TMSG-1 specific and is effective for Western blot and immunohistochemistry assays to detect TMSG-1 expression in cancer cells. TMSG-1 protein is about 45 000 (relative molecular mass) at cell membrane and cytoplasm of tumor cells. Expression of TMSG-1 protein correlates well, inversely with the tumor metastatic potential.
Zhonghua bing li xue za zhi Chinese journal of pathology 01/2005; 34(1):15-21.
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ABSTRACT: To explore the significance of BCL-10 protein expression in the gastrointestinal mucosa-associated lymphoid tissue (MALT) lymphoma.
Immunohistochemistry studies were performed using CD20, CD79a, CD3, CD45RO, CD23, CD5, CD10 monoclonal antibodies in 43 cases of gastrointestinal MALT lymphomas, including 25 indolent classical MALT lymphomas and 18 MALT lymphomas with large cell transformation. BCL-10 protein expression was assayed in the tumor cells.
In 25 low-grade MALT lymphomas, expression of BCL-10 was found in the nuclei in 10 cases, in both nuclei and cytoplasm 1 case, in cytoplasm 3 cases and no expression 11 cases. In 18 transformed MALT lymphomas, BCL-10 was expressed in the nuclei in 7 cases, in both nuclei and cytoplasm 1 case, in cytoplasm 2 cases, no expression 8 cases. The frequency of BCL-10 expression in nuclei was the highest (44.2%).
The frequency of BCL-10 expression in nuclei in the gastrointestinal MALT lymphoma is high, indicating that it may associate with the pathogenesis of this entity, and may be helpful to its diagnosis.
Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi 11/2004; 25(10):592-5.
