[Mutation of isocitrate dehydrogenase gene in Chinese patients with glioma].

OBJECTIVE: To investigate mutation status of isocitrate dehydrogenase (IDH) 1 and IDH2 genes in Chinese patients with gliomas in correlation with clinicopathological characteristics. METHODS: Formalin-fixed and paraffin-embedded (FFPE) tissue samples of 234 gliomas were collected including the matched blood samples in 30 patients. DNA was extracted, followed by PCR-Sanger sequencing to detect IDH1 and IDH2 gene mutations. Immunohistochemistry was performed using mutation-specific antibody recognizing IDH1R132H mutation. Immunostains for p53 and epidermal growth factor receptor (EGFR) were also performed. Oligodendroglial tumors with IDH mutation were double stained with IDH1R132H and GFAP by immunofluorescence to investigate the location of IDH1R132H expression. RESULTS: (1) By IDH1 heterozygous somatic mutation analysis, Arg132His (c: G395A) was found in 31.6% (74 of 234) of the cases. IDH mutations were more frequent in oligoastrocytomas (9/13), anaplastic oligoastrocytomas (7/11), oligodendrogliomas(18/26, 69.2%), anaplastic oligodendrogliomas (8/10), and less frequent in diffuse astrocytomas (17/47, 36.2%), anaplastic astrocytomas (5/18), and glioblastomas (10/69, 14.5%). The mutation rate inversely correlated with the tumor grade in a linear fashion in astrocytic tumors (P = 0.007). Primary glioblastomas were characterized by a lower frequency of mutations than secondary glioblastomas (5/55 vs. 5/14, P = 0.036); IDH mutation was not detected in pilocytic astrocytoma and ependymoma. No IDH2 mutation was identified in this study cohort. (2) Immunohistochemistry of IDH1R132H demonstrated a strong cytoplasmic staining in 80 cases, which was highly correlated with IDH mutation status (P = 0.001). IDH1R132H was highly specific to tumor cells. (3) p53 immunostain was significantly correlated the IDH mutation in diffuse astrocytoma, anaplastic astrocytoma and secondary glioblastomas (P = 0.007, 0.026, 0.038 respectively). (4) No correlation between EGFR and IDH mutation was found. CONCLUSIONS: High prevalence of IDH heterozygous somatic mutation occurs in the earlier stage of gliomas, which can be detected by mutation-specific antibody IDH1R132H. Furthermore, evaluation of p53 and EGFR expression combined with IDH mutation analysis may significantly aid in the diagnosis and differential diagnoses of gliomas in Chinese patients.

[Insulin protects the loss of colonic interstitial cells of Cajal and acetylcholine in patients with type 2 diabetes mellitus].

OBJECTIVE: A deficiency of interstitial cells of Cajal (ICC) and neuronal nitric oxide synthase (nNOS) in the gastrointestinal muscle layer have been shown in both diabetic animal models and patients with diabetes mellitus (DM), but little is known about the alteration of colonic acetylcholine (Ach) expression in patients with DM, and it is remain unclear whether those changes are related to different treatments. This study examined whether the ICC density, or the expression of Ach containing nerves in the colon, was altered in patients with type 2 DM, and whether those changes were protected by insulin treatment. METHODS: Paraffin embedded colonic specimens that have been fixed in 10% formalin were collected from 81 patients with colon cancer who underwent colectomy (non-DM as controls, M/F = 17/12; DM with insulin treatment, M/F = 12/12; DM with oral medication treatment, M/F = 19/9). Serial sections were stained with antibodies to c-kit and Ach using immunohistochemical method.Meanwhile, mast cells were stained with toluidine blue. The ICC number was calculated by subtraction of the number of mast cells from the number of c-kit positive cells. The number of ICC and the expression of Ach were compared among the controls and DM patients with different treatment. RESULTS: C-kit positive cells, which located within the inner muscle layer and muscular plexus mainly, were expressed in control group and they were much more regular and less vacuolization than that of controls. The density of both intramuscular-ICC (ICC-IM) and myenteric ICC (ICC-MY) were much lower in patients with DM than that in the controls. Furthermore, ICC density in DM patients who underwent insulin treatment was higher than that in patients with oral medical therapy (control vs insulin vs oral medication group: ICC-MY, 60.12 vs 23.95 vs 16.49, P = 0.000; ICC-IM, 41.79 vs 33.18 vs 25.88, P = 0.000). In addition, the alteration of Ach expression in colonic enteric nerves was similar as the pattern of ICC (control vs insulin vs oral medication group: 147.50 vs 103.82 vs 86.38, P = 0.000). There was no influence of gender and age to the alteration of ICC and Ach. CONCLUSIONS: The colonic ICC and Ach in DM patients is irregular, and the distribution is loose, with lots of vacuolization. The density of clonic ICC-IM and ICC-MY were lower in DM patients than that in controls accompanied with downregulation of enteric nerves Ach expression. These alterations could be protected by insulin treatment.

[Improvement and application of diagnostic techniques of the skeletal muscle biopsy].

OBJECTIVE: To summarize the improvement of various muscle staining techniques and discuss their application in diagnosis of neuromuscular diseases. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Investigation in vivo of effect of ultrasound-mediated microbubble destruction on entrance of feridex into the aortal wall].

OBJECTIVE: To explore feasibility for entrance of the contrast agent Sonovue and Feridex into the aortal wall. METHODS: 17 male Japanese giant ears rabbits (common grade), including 11 atherosclerosis (AS) animal models fed with food containing high-content lipid and normal animals fed with common food as control. Respectively, 10 animals in the AS group and 6 animals in the normal group were selected in a random way to undergo ultrasound-mediated microbubble destruction (UMMD) and no ultrasound-mediated microbubble destruction (-UMMD) half and half. One animal was administrated with double doses of Feridex. After general anesthesia, MR plain scan and intravenous injection of Feridex 100 micromol Fe/kg, immediately ultrasound focused on the front wall of the aortic arch, which underwent UMMD at the pressure of 3.5 Mpa with MI1.2 while 10 ml solution (Sonovue + normal saline)was injected intravenously at the speed of 0.5 ml/min FOR 20 min. 3T magnetic resonance (MR) was performed with a moderately T2* weighted gradient sequence. Enhanced scan were performed for 1 h, 24 h, 48 h, 72 h and after killing the animal. then the specimen were delivered to conduct optical and electronic microscope examination. Variance test for the re-measured data was adopted to verify the data obtained in every group. RESULTS: The effect of UMMD group on SPIO particles entrance into the aortal wall is of marked significance (P = 0.0004) statistically. The effect of UMMD on distribution in the vessel wall is of statistical significance (P = 0.01), more particles in the dventitia. Gas or microbubbles were found to enter into the intima, media of the aorta, and verified by Oil Red O staining. After staining the findings of iron particle in the cell and out of the cell are different. CONCLUSIONS: UMMD may facilitate entrance of those SPIO particles with a bigger diameter and microbubbles into the aortal wall. This discovery may provide a new solution for penetration of complex macromolecule probes and gene-carried drug through the tunica intima of the aorta.

[Pathology of enterovirus 71 infection: an autopsy study of 5 cases].

OBJECTIVE: To study the clinicopathologic features of fatal enterovirus 71 (EV71) infection. METHODS: Autopsy was performed in 5 neonates died of EV71 infection. Tissue samples from major organs were collected, formalin-fixed and examined under light microscopy. Immunohistochemical study was carried out in selected examples. RESULTS: Four of the 5 cases showed predominant changes in central nervous system, with encephalitis and encephalomyelitis identified mainly in brainstem and upper cervical spinal cord. Histologic findings included neuronal degeneration and necrosis, neuronophagia, perivascular cuffing and diffuse or nodular hyperplasia of macrophages/microglia. Cerebral edema, brain herniation and aseptic meningitis were also noted. The lungs showed mainly pulmonary congestion, neurogenic pulmonary edema and focal hemorrhage. There were minimal changes in the intestinal epithelium. The intestinal lymphoid tissue however was hyperplastic and associated with apoptosis of follicular center cells. The remaining case had cerebral edema and mild meningitis. The lung alveolar septa were thickened with lymphocytic infiltrates. Some alveolar cells were hyperplastic and associated with diffuse hyaline membrane formation. No specific abnormalities were identified in gastrointestinal tract. In all the 5 cases studied, there was enlargement of lung hilar and mesenteric lymph nodes, coupled with apoptosis of follicular center cells. In general, no significant pathologic changes were demonstrated in heart, liver and kidneys. CONCLUSIONS: In fatal EV71 infection, the major pathologic changes lie in the central nervous system. The pulmonary lesions are mainly secondary in nature. The usual cause of death is cerebral edema complicated by brain herniation and pulmonary edema. It is also noteworthy that some cases show only lung damages, without classic neurologic changes.

[Muscle atrophy of hand as an initial sign in a patient with protoplasmic astrocytoma: a case report and literature review].

We reported a rare case of protoplasmic astrocytoma presenting small muscle atrophy of the right hand as an initial sign. A 39-year-old male was admitted to hospital complaining of chronic muscle atrophy and subtle headache. Electromyography (EMG) showed brief small denervation and no signs of sensory-motor conduction impairment. CT and MRI revealed multiply expansive intracranial lesion in left hemisphere, which was highly suspected of cerebral echinococcus or Balo disease. The patient underwent surgical excision and pathological report was protoplasmic astrocytoma, with glial fibrillary acidic protein (GFAP, +++) of immunohistochemical method. We reviewed clinical features, radiological manifestations and pathology of protoplasmic astrocytoma with medical literature documents.

Expression of E-cadherin, beta-catenin, cathepsin D, gelatinases and their inhibitors in invasive ductal breast carcinomas.

BACKGROUND: E-cadherin, beta-catenin, cathepsin D, matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 are all invasion-related proteins. The expression patterns of these proteins in invasive ductal breast carcinomas, and their associations with known clinicopathological parameters, tumor recurrence and expressions of estrogen receptor (ER), progesterone receptor (PR), PS2 and c-erbB2 were not well studied in Chinese patients. METHODS: In a set of 94 invasive ductal breast carcinomas, protein expressions of these molecular markers were investigated by immunohistochemistry, and their associations with known clinicopathological parameters, tumor recurrence and expressions of ER, PR, PS2 and c-erbB2 were also examined. In addition, the interrelationship between the expressions of these proteins were studied. RESULTS: Preserved membrane E-cadherin expression was associated with late tumor stage and tumor recurrence, whereas the reduced junctional beta-catenin associated with positive lymph node status and c-erbB2 overexpression. Positive staining of cathepsin D in tumor stromal cells displayed a significant association with late tumor stage. High expression of MMP-2 in cancer cells was associated with large tumor size and PR positive expression. TIMP-2 expression was positively associated with tumor recurrence. In addition, inter-relationship between the expressions of these biomarkers was also assessed. Cathepsin D staining in cancer cells was inversely correlated with its staining in stromal cells, and also inversely correlated with MMP-2 staining in tumor stromal cells. MMP-2 expression in stromal cells displayed an inverse correlation with TIMP-2 expression. MMP-9 expression displayed parallel associations with TIMP-1 and TIMP-2 expression. CONCLUSION: Evaluation of E-cadherin, beta-catenin, cathepsin D, MMP-2 and TIMP-2 expression may be of some help in more accurately predicting the prognosis of invasive ductal breast carcinomas.

[Inhibiting cerebral glioma growth with continuous low-dose chemotherapy and cyclooxygenase-2 inhibitor in nude mice].

OBJECTIVE: To observe the changes in glioma growth characteristic and apoptosis of tumor cells after single handed continuous low-dose chemotherapy, cyclooxygenase-2 inhibitor treatment alone and the combination of the two treatments. METHODS: The U251MG cells in exponential phase of growth were made into 10(7)/mL cell suspension in free-serum 1640 and stored in 37 degrees C incubator. The survival rate of cells was above 95%. The U251MG cells were implanted into the right parietooccipital lobe of the 4-week old nude mouse with a 5 micro liter micro amount sample injector. The number of injected U251MG cells was 5 x 10(4) for a mouse. Twenty days after the model making, the nude mice were treated with elemene and indometacin respectively and the combination of them, twice a week. The mice were divided into four groups. Group I was treated with indometacin alone, group II elemene alone, group III low-dose elemene plus indometacin, Group IV was used as controls, including tumor control and blank control. The animals were killed on the 40th and 50th day after implantation by breaking cervical vertebra. The fixed brain was made into 3 microm slices by paraffin section. The slices were carried out with HE staining and immunohistochemical staining of glial fibrillary acidic protein(GFAP), cell proliferation-associated antigen(Ki-67), cyclooxygenase-2(COX-2), CD34, programmed cell death 5(PDCD5) and terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling(TUNEL). RESULTS: The proliferation of glioma cells was predominant in the tumor control mouse brain. Several immature blood vessels were observed in the tumor implanted for 40 days. The white matter was infiltrated by bulk glioma cells along with capillary vessel clusters in the mouse brain implanted for 50 days. In groups with combination treatment of the two drugs, 40 days after the implantation, several apoptosis cells and glioma cells were observed in tumor where the positive signal for GFAP was showed; and 50 days after the implantation, lots of apoptosis cells were observed in tumor cell implantation area where the negative signal for GFAP and positive signal for PDCD5 was showed. The volume of tumor was (29.8+/-39.1) mm(3) 40 days after the implantation, and (78.4+/-125.9) mm(3) 50 days after the implantation. There was no statistically significant difference in tumor volume among groups(P=0.11). CONCLUSION: The combination of two treatments could merely prolong the survival time of the nude mouse model, without the effect of eliminating the tumor completely.

[Effects of beta1-integrin, fibronectin and laminin on invasive behavior of human gliomas].

OBJECTIVE: To investigate the effects of beta1-integrin, fibronectin (FN) and laminin (LN) on the invasive behavior of human gliomas. METHODS: Functional impacts of beta1-integrin, fibronectin and laminin on cell adhesion, migration and metastasis of U251 malignant glioblastoma cells were investigated by in vitro adhesion, migration and invasion assays. The amount and distributions of cellular microfilaments and pseudopodia were studied by fluorescent cytochemistry, confocal laser scanning microscope and scanning electron microscope. Lastly, beta1-integrin, fibronectin and laminin were investigated for their roles in cellular microfilament skeleton. RESULTS: (1) Fibronectin did not affect cell adhesion of U251MG cells, but anti-beta1 integrin antibodies inhibited cell adhesion (P < 0.01); Laminin stimulated cell adhesion of U251MG cells (P < 0.01) but anti-beta1 integrin antibodies had little effect on the laminin-mediated cell adhesion. (2) The migration of U251MG cells on dishes coated with FN was inhibited by anti-beta1 integrin antibodies (P < 0.05). (3) F-actins formed strong and dense stress fibers in U251MG cells on dishes coated with FN and LN. Anti-beta1 integrin antibodies disrupted the microfilament network and F-actin aggregation. (4) FN and LN increased the number of pseudopodia on cell surface, whereas anti-beta1 integrin antibodies reversed this function. (5) FN and anti-beta1 integrin antibodies had little effects on the invasive ability of U251MG cells in vitro. The invasion was increased by LN, but inhibited by anti-beta1 integrin antibodies. CONCLUSIONS: (1) The interaction between beta1-integrin, FN may stimulate U251MG cell migration via changing the structures of microfilament skeleton and the number of pseudopodia. (2) beta1-integrin may play a role in the LN-mediated in vitro invasion of U251MG cells.

ATRX loss in adult supratentorial diffuse astrocytomas correlates with p53 over expression and IDH1 mutation and predicts better outcome in p53 accumulated patients.

BACKGROUND: IDH1/2 mutation, 1p/19q-codeletion and MGMT hypermethylation are well known molecular markers for gliomas. ATRX and p53 alterations are two lineage-specific genetic aberrations in diffuse astrocytic tumors. The aim of the present study is to clarify the significance of ATRX loss and its correlation with p53 overexpression, IDH1/2 mutations, 1p/19q-codeletion and MGMT hypermethylation in supertentorial astrocytoma, and to determine the prognostic value of these factors in Chinese patients. METHODS AND RESULTS: A total of 135 adult supertentorial astrocytomas were evaluated. ATRX loss was detected by immunohistochemistry (IHC) and was shown to be much less frequent in pGBs (3.5%) than in grade II, III astrocytomas and IV sGBs (31%). Direct sequencing and/or IHC analysis of the IDH1R132H gene mutation and p53 accumulation demonstrated correlation with age. Strong correlations were found between ATRX loss and IDH1R132H mutation, p53 overexpression as well as MGMT hypermethylation. 1p/19q-codeletion detected by fluorescence in situ hybridization (FISH) showed mutually exclusive with ATRX loss and p53 accumulation. In addition, patients with p53 overexpression combined with ATRX alterations demonstrated substantially longer survival than patients with wild-type ATRX. CONCLUSIONS: There may be interactions among these distinct molecules in astrocytoma development. ATRX loss may predict better clinical outcome in astrocytoma patients with p53 overexpression as compared to patients with wild-type ATRX. Tumors with astrocytoma phenotype accompanied by 1p/19q-codeletion and IDH1R132H mutation are mutually exclusive with ATRX and p53 alterations. Routine IHC can be used for evaluation of ATRX loss, p53 protein accumulation and IDH1R132H mutation, which may allow a means of classification of astrocytoma outcome.

[Clinical pathological study of the anterior lens capsule abnormalities in familial congenital aniridia with cataract].

OBJECTIVE: To observe the pathological changes of the lens and anterior lens capsule of the patients with familial congenital aniridia, and discuss the histopathological etiology of the fragility of the anterior capsule and the significance of surgical project. METHODS: Anterior lens capsules and lens specimens were obtained from aniridic patients during cataract surgery. The intraoperative behavior of each capsule was noted, after which the specimens were submitted for histopathologic evaluation and electron microscope examination. RESULTS: The anterior lens capsule was extremely fragile and remarkably thin. Degenerative changes(degeneration, necrosis, loss) of the lens epithelium and discontinuity of the lens epithelium were found in some specimens. Proliferation and double layer of the epithelial cells in some area of the specimens can be seen also. Ply structure of the anterior capsule became thin or disappeared. CONCLUSION: Degenerative or proliferative changes of the lens epithelial cells were associated with the thinness and extreme intraperative fragility of the anterior lens capsules in familial aniridia with cataract. Greater awareness of anterior capsule fragility in some aniridic patients with cataract may reduce the risk of capsule complications and lead to safer surgical outcomes.

[Expression of SARS-CoV in various types of cells in lung tissues].

OBJECTIVE: To investigate the cell types infected by severe acute respiratory syndrome-associated coronavirus (SARS-CoV) in lung tissues and explore the mechanism of lung injury in SARS. METHODS: In-situ hybridization(ISH) and immunohistochemistry(IHC) double staining was applied to study the lung tissues from 7 SARS cases of Beijing and one of Anhui province. According to SARS-CoV genome sequence, the cDNA probe was synthesized and labelled by digoxin. Immunohistochemically, antibodies of cytokeratin(CK), CD34, CD68, Vimentin and CD3 were applied to demonstrate bronchial epithelial cells, type II pneumocytes, endothelial cells, macrophages, fibroblasts and T cells respectively. RESULTS: The positive results of in-situ hybridization showed that the lung tissues of all cases expressed SARS-CoV RNA, and positive signals displayed in cytoplasms (purple-blue, NBP-BCIP. ISH-IHC double staining showed that positive signals of both ISH (purple-blue NBT-BCIP and IHC (red-brown, AEC expressed in the cytoplasms (purple and red). The positive results of double staining indicated that bronchial epithelial cells, type II pneumocytes, endothelial cells, macrophages, fibroblasts and T lymphocytes were diffusely infected by SARS-CoV. CONCLUSION: This study of ISH-IHC double staining in lung tissues of SARS patients showed that bronchial epithelial cells, type II pneumocytes, endothelial cells, macrophages, T lymphocytes and fibroblasts were attacked diffusely in SARS lungs. Various types of cells damaged by SARS-CoV and inflammatory mediators released by those cells play an important role in the pathogenesis of lung injury in SARS.

[Lung pathology and pathogenesis of severe acute respiratory syndrome: a report of six full autopsies].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Nerve biopsy findings contribute to diagnosis of multiple mononeuropathy: 78% of findings support clinical diagnosis.

Multiple mononeuropathy is an unusual form of peripheral neuropathy involving two or more nerve trunks. It is a syndrome with many different causes. We reviewed the clinical, electrophysiological and nerve biopsy findings of 14 patients who suffered from multiple mononeuropathy in our clinic between January 2009 and June 2013. Patients were diagnosed with vasculitic neuropathy (n = 6), perineuritis (n = 2), chronic inflammatory demyelinating polyradiculoneuropathy (n = 2) or Lewis-Sumner syndrome (n = 1) on the basis of clinical features, laboratory data, electrophysiological investigations and nerve biopsies. Two patients who were clinically diagnosed with vasculitic neuropathy and one patient who was clinically diagnosed with chronic inflammatory demyelinating polyradiculoneuropathy were not confirmed by nerve biopsy. Nerve biopsies confirmed clinical diagnosis in 78.6% of the patients (11/14). Nerve biopsy pathological diagnosis is crucial to the etiological diagnosis of multiple mononeuropathy.

[Pathologic study of circulating blood leukocytes in severe acute respiratory syndrome].

OBJECTIVE: To study the pathologic characteristics and pathogenesis of circulating blood leucocytes infected by severe acute respiratory syndrome associated coronavirus (SARS CoV or SCV) in SARS patients. METHODS: Blood samples of 22 SARS patients were studied, and 4 healthy blood samples were observed as negative controls. The white blood cells were collected from whole blood. The ultrastructural characteristics were observed by transmission electron microscopy. CD45RO antibody was used for pre-embedding immunoelectron microscopy. The SARS viral sequence was detected with real-time polymerase chain reaction (RT-PCR). RESULTS: Coronavirus-like particles were founded in the leukocytes in 6 of the 22 blood samples. Five of them gave positive results in the real-time PCR. The number of granulocytes was increased (P < 0.05) and that of lymphocytes was decreased (P < 0.05) respectively. Immunoelectron microscopy showed that CD45RO positive T lymphocyte decreased to 6% - 7%. Circulating lymphocytes had the highest percentage of infection. The morphologic characteristics of coronavirus-like particles were spherical or oval in shape, about 80 - 120 nm in diameter, with a dense round core and a clear halo around the core. A distinct membrane and club-shaped surface projections were seen in the periphery. The particles were located in the cytoplasm, the cisternae of endoplasmic reticulum, Golgi apparatus and vesicles. Virus entered cells by endocytosis or membrane fusion and was released through a budding process. CONCLUSION: Our data suggested that lymphocytes, particularly T cells, were probably the target cells of SARS CoV. The viruses may actively infected the immune cells during SARS CoV acute infection phase and the destruction of target cells may be one of the important reasons for the death of the circulating leukocytes in SARS.

[Expression and significance of integrin alpha6beta4 in experimental allergic neuritis].

OBJECTIVE: To investigate the expression of the integrin alpha6beta4 in experimental allergic neuritis (EAN) and the relationship of the integrin alpha6beta4 with functional states of Schwann cells (Sc) as well as the injury and repair of the myelin during EAN. METHODS: EAN was induced in Lewis rats and sciatic nerves were resected in 18 EAN and 3 normal rats. The expression of tissue integrin alpha6beta4 was analyzed during the course of EAN induction and in controls by in situ hybridization and semi-quantitative RT-PCR. RESULTS: The detection of integrin alpha6 and beta4 subunit by hybridization in situ demonstrated that expression of alpha6 subunit present no significant changes during the course of EAN, while expression of beta4 declined in the early phase, showing less positive signals than those of the control, and restored its expression in the later or recovery phase. The changes of expression of integrin alpha6 and beta4 in EAN were confirmed by semi-quantitative PT-PCR, using GAPDH as the internal standard. CONCLUSIONS: The degeneration and injury of Sc caused by inflammation affect the expression of integrin, which shows similar changes in Sc during embryogenesis, indicating alpha6beta4 may be a marker of Sc differentiation and at least an important molecule to mark the course of EAN. The expression of alpha6beta4 correlate with the injury and repair of myelin during EAN.

Intraneural perineurioma affecting multiple nerves: a case report and literature review.

Intraneural perineurioma is a neoplasm of perineurial cells, corresponding to WHO grade I. We present a case of intraneural perineurioma affecting multiple nerves, which usually involved one or two of major nerve trunks in one patient. We describe the clinical presentation, magnetic resonance (MR) neurography characteristics, and pathological characteristics. The differential diagnosis with other diseases, such as neurofibroma, Schwannomatosis and HNPP, will also be discussed. We also review the literature in efforts to highlight recent studies on intraneural perineurioma and heighten and awareness for the possible presentations of this disorder.