Adolescent ; Adult ; Aspergillosis ; complications ; microbiology ; Aspergillus ; isolation & purification ; Candida albicans ; isolation & purification ; Candidiasis ; complications ; microbiology ; Female ; Follow-Up Studies ; Granulomatosis with Polyangiitis ; complications ; microbiology ; pathology ; Humans ; Male ; Middle Aged ; Mucor ; isolation & purification ; Mucormycosis ; complications ; microbiology ; Nocardia ; isolation & purification ; Nocardia Infections ; complications ; microbiology ; Retrospective Studies ; Young Adult

OBJECTIVETo study the promoter methylation pattern of p16 and hMLH1 genes in esophageal squamous cell carcinoma and reflux esophagitis, and to correlate the results with clinical and pathologic findings.

METHODSTwelve cases of normal esophagus, 13 cases of esophageal squamous cell carcinoma, 43 cases of reflux esophagitis with basal cell hyperplasia and 21 cases of reflux esophagitis with dysplasia, as confirmed by endoscopic and pathologic examination, were enrolled into the study. Genomic DNA was extracted. The promoter methylation status of p16 was measured by methylation-specific polymerase chain reaction. The promoter methylation status of hMLH1 was measured by sodium bisulfite-restriction enzyme digestion. Immunohistochemical study for p16 and hMLH1 proteins was also carried out.

RESULTSThe rates of p16 methylation in normal esophageal epithelium, basal cell hyperplasia, dysplasia and esophageal squamous cell carcinoma were 0/12, 14.0% (6/43), 38.1% (8/21) and 6/13, respectively. The p16 methylation correlated with the progress of esophageal lesions. On the other hand, the hMLH1 methylation was not observed in the normal esophageal epithelium and reflux esophagitis. One case of esophageal squamous cell carcinoma showed the presence of hMLH1 methylation. The hMLH1 promoter hypermethylation did not correlate with the clinical and pathologic features.

CONCLUSIONSThe p16 methylation may be one of the earliest events in the pathogenesis of esophageal squamous cell carcinoma and is also observed in reflux esophagitis. Reflux esophagitis may be related to the development of esophageal squamous cell carcinoma in Chinese population. In contrast, hMLH1 methylation may not be directly involved in the tumorigenesis of esophageal squamous cell carcinoma.

Adaptor Proteins, Signal Transducing ; genetics ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; DNA Methylation ; Esophageal Neoplasms ; genetics ; pathology ; Esophagitis, Peptic ; genetics ; pathology ; Esophagus ; pathology ; Female ; Genes, p16 ; Humans ; Hyperplasia ; Male ; Middle Aged ; MutL Protein Homolog 1 ; Nuclear Proteins ; genetics ; Precancerous Conditions ; genetics ; pathology ; Promoter Regions, Genetic ; genetics

OBJECTIVETo differentiate between Aspergillus species and Mucorales of fungal sinusitis by immunohistochemistry.

METHODSFormalin-fixed paraffin-embedded tissue blocks of 66 cases of fungal sinusitis were retrieved from the archival files of Department of Pathology of Beijing Tongren Hospital during the period from 2001 to 2006. The samples included 29 cases of fungal balls, 12 cases of allergic fungal sinusitis, 24 cases of chronic invasive fungal sinusitis and 1 case of acute invasive fungal sinusitis. The types of fungi were 44 Aspergillus species (31 cases of A. fumigatus, 7 cases of A. flavus and 6 cases of A. terreus) and 22 Mucorales (14 cases of Mucor species and 8 cases of Rhizopus species). Immunohistochemistry was performed with MUC2, MUC5AC and MUC5B antibodies. The results were compared with histochemical study for periodic acid-Schiff (PAS) and Grocott methenamine silver (GMS) stains.

RESULTSImmunohistochemical study for MUC5B showed that the positive rate of Aspergillus species was 90.9%, in contrast to 4.5% in Mucorales (P < 0.001). The expression of MUC2 and MUC5AC was completely negative, whereas PAS and GMS stains were positive in all cases.

CONCLUSIONMUC5B antibody appears to be a useful immunohistochemical marker for identifying fungal types in tissue sections, especially in distinguishing between Aspergillus species and Mucorales in fungal sinusitis.

Antibodies, Fungal ; immunology ; Antibody Specificity ; immunology ; Aspergillosis ; diagnosis ; immunology ; Aspergillus flavus ; immunology ; Aspergillus fumigatus ; immunology ; Diagnosis, Differential ; Humans ; Immunohistochemistry ; methods ; Mucin-5B ; genetics ; immunology ; Mucor ; immunology ; Mycoses ; diagnosis ; immunology ; microbiology ; Sinusitis ; diagnosis ; microbiology

AIM: To investigate the effects of baicalein(BAI)on the proliferation and migration of gastric cancer MGC-803 cells and the mechanisms.METHODS:After MGC-803 cells were treated with BAI at different concen-trations,the viability of the MGC-803 cells was tested by MTT assay.The cell colony formation ability were detected by plate colony formation assay.Wound-healing and Transwell cell migration assays were used to test the migration ability of the MGC-803 cells.The concentration of 12-hydroxyeicosatetraenoic acid(12-HETE)was measured by ELISA.The pro-tein levels of platelet type 12-lipoxygenase(p12-LOX),vascular endothelial growth factor(VEGF),p-ezrin and epithelial-mesenchymal transition(EMT)markers in MGC-803 cells were determined by Western blot.RESULTS:BAI significantly inhibited the proliferation,plate colony formation and migration abilities of the MGC-803 cells(P<0.05 or P<0.01), down-regulated the concentration of p12-LOX metabolite 12-HETE significantly(P<0.05 or P<0.01), decreased the protein levels of p12-LOX,VEGF,p-ezrin,vimentin and Snail(P<0.05 or P<0.01),and increased the protein expres-sion of E-cadherin(P<0.01).CONCLUSION:BAI suppresses the proliferation and migration abilities of gastric cancer MGC-803 cells effectively.These effects of BAI may be related to regulating the protein levels of p12-LOX,VEGF,p-ezrin and EMT-related proteins.

Toxoplasma gondii infections occur throughout the world, and efforts are needed to develop various vaccine candidates expressing recombinant protein antigens. In this study, influenza matrix protein (M1) virus-like particles (VLPs) consisting of T. gondii rhoptry antigen 4 (ROP4 protein) were generated using baculovirus (rBV) expression system. Recombinant ROP4 protein with influenza M1 were cloned and expressed in rBV. SF9 insect cells were coinfected with recombinant rBVs expressing T. gondii ROP4 and influenza M1. As the results, influenza M1 VLPs showed spherical shapes, and T. gondii ROP4 protein exhibited as spikes on VLP surface under transmission electron microscopy (TEM). The M1 VLPs resemble virions in morphology and size. We found that M1 VLPs reacted with antibody from T. gondii-infected mice by western blot and ELISA. This study demonstrated that T. gondii ROP4 protein can be expressed on the surface of influenza M1 VLPs and the M1 VLPs containing T. gondii ROP4 reacted with T. gondii-infected sera, indicating the possibility that M1 VLPs could be used as a coating antigen for diagnostic and/or vaccine candidate against T. gondii infection.
Animals ; Baculoviridae ; Blotting, Western ; Clone Cells ; Cloning, Organism ; Enzyme-Linked Immunosorbent Assay ; Influenza, Human* ; Insects ; Mice ; Microscopy, Electron, Transmission ; Toxoplasma* ; Toxoplasmosis ; Virion

OBJECTIVETo characterize clinicopathological features of allergic fungal sinusitis (AFS).

METHODSThirty-six cases of AFS were retrieved from the department archival files of Beijing Tongren Hospital from 2002 to 2006. AB-PAS, GMS and MUC5B stain were performed using paraffin-embedded tissues of the cases. Ten cases with available fresh diagnostic tissue were investigated by electron microscopy.

RESULTSPatients included 21 males and 15 females. The age of patients ranged from 11 to 53 years. Atopy was very common in these patients. On plain CT scans, the affected nasal sinuses were filled with soft tissue shadow with patchy hyperdensity. The bony sinus wall showed areas of pressure erosion. Skin antigen tests showed fungal positivity in 31 of 36 cases. Serum levels of the total IgE and/or the specific fungal IgE were elevated in 20 cases. The eosinophil quantity was elevated in 23 cases. Fungal culture was positive in 10 cases. Gross examination showed thick putty secretions within the lesions. Light microscopy showed typical "eosinophilic mucin". Fungal elements were seen with AB-PAS, GMS and MUC5B stains. Electron microscopy demonstrated degranulation by the eosinophils.

CONCLUSIONS"Eosinophilic mucin" is the typical histopathological feature of AFS. AB-PAS, GMS and MUC5B staining methods can used to detect fungal species in mucin. Accurate diagnosis of AFS requires correlations among clinical findings, radiologic examinations, laboratory tests and histopathologic features. However, the ultimate diagnosis requires a histopathologic confirmation.

Adolescent ; Adult ; Child ; Eosinophils ; microbiology ; ultrastructure ; Female ; Fungi ; isolation & purification ; ultrastructure ; Humans ; Hypersensitivity ; blood ; immunology ; pathology ; Immunoglobulin E ; blood ; Leukocyte Count ; Male ; Middle Aged ; Paranasal Sinuses ; diagnostic imaging ; microbiology ; pathology ; Radiography ; Sinusitis ; blood ; immunology ; microbiology ; pathology ; Young Adult

OBJECTIVETo compare the differences in clinicopathologic features of invasive fungal rhinosinusitis caused by Aspergillus and Mucorales, and to discuss the pathogenesis of tissue injury induced by these two kinds of fungi.

METHODSThe clinical and pathologic features of 19 patients with invasive fungal rhinosinusitis due to Aspergillus (group A) and 16 patients with invasive fungal rhinosinusitis due to Mucorales (group M) were retrospectively reviewed. HE, PAS and GMS stains were performed on all the paraffin-embedded tissues. The diagnosis was confirmed by histologic examination and microbiological culture results.

RESULTSAmongst the group A patients, the clinical course was acute in 4 cases and chronic in 15 cases. Thirteen cases had underlying predisposing conditions, including diabetes (number = 4), malignant tumor (number = 5), history of trauma (number = 1) and radical maxillary sinus surgery (number = 3). Follow-up information was available in 13 patients. Seven of them died, 4 due to fungal encephalopathy and 3 due to underlying diseases. Amongst the group M patients, the clinical course was acute in 14 cases and chronic in 2 cases. Fourteen cases had underlying predisposing conditions, including diabetes (number = 8), malignant tumor (number = 5) and history of wisdom tooth extraction (number = 1). Follow-up information was available in 14 patients. Four of them died of fungal encephalopathy. There was significant difference in clinical onset between the two groups (P = 0.01). There was however no difference in terms of underlying predisposing conditions and disease mortality. Histologically, the microorganisms in group A patients formed fungal masses and attached to the mucosal surface, resulting in necrotic bands (11/19). Epithelioid granulomas were conspicuous but multinucleated giant cells were relatively rare. Deep-seated necrosis, granulomatous inflammation against fungal organisms (3/19) and vasculitis with thrombosis (4/19) were not common. On the other hand, large areas of geographic necrosis involving deep-seated tissue could be seen in group M patients (13/16). Isolated multinucleated giant cells were commonly seen. Granulomatous inflammation against fungal organisms were identified (16/16). Vasculitis and thrombosis were also observed (10/16).

CONCLUSIONSThe invasiveness of Mucorales is remarkable; and when it causes invasive fungal rhinosinusitis, the clinical course is often acute and large areas of tissue necrosis can be seen. The invasiveness of Aspergillus in tissue is relatively mild. Granulomas are more common and the disease often runs a chronic clinical course. There is however no significant difference in long-term mortality. The pathogenesis may be related to the different components of the fungi.

Adolescent ; Adult ; Aged ; Aspergillosis ; diagnosis ; microbiology ; pathology ; Aspergillus ; isolation & purification ; pathogenicity ; Child ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Mucorales ; isolation & purification ; pathogenicity ; Mucormycosis ; diagnosis ; microbiology ; pathology ; Retrospective Studies ; Rhinitis ; diagnosis ; microbiology ; pathology ; Sinusitis ; diagnosis ; microbiology ; pathology ; Young Adult

OBJECTIVETo study the clinicopathologic features of malignant tumors in head and neck region complicated by fungal infection.

METHODSTwenty-one cases of malignant tumors occurring in head and neck region complicated by fungal infection were retrieved from the archival file. The light microscopic findings were reviewed. Histochemical (for PAS and GMS) and immunohistochemical (for MUC5B) studies were carried out. Fungal culture results were available in 13 of the 21 cases.

RESULTSThe age of the patients ranged from 12 to 72 years (median = 48 years). The male-to-female ratio was 17:4. Eight cases (38.1%) were complicated by invasive fungal sinusitis, with orbital involvement in 6 cases and brain involvement in 1 case. The primary tumors in such cases included leukemia (n = 7) and nasopharyngeal carcinoma (n = 1). The fungi belonged to Zygomycete in 5 cases and Aspergillus in 3 cases. These patients had history of chemotherapy/radiotherapy or antibiotics usage. The remaining 13 cases of fungal infection often affected necrotic tumor tissue in nasal cavity, paranasal sinuses, pharynx, larynx and palate. The fungi involved were Aspergillus (n = 6) and Candida (n = 4). Seven of such patients had received radiotherapy. Fungal culture was positive in 9 cases. Fourteen patients had follow-up information available and six of them died of the disease.

CONCLUSIONSMalignant tumors occurring in head and neck region can be complicated by fungal infection. Invasive fungal sinusitis (due to Zygomycetes and Aspergillus) often occurs in patients with leukemia, tends to involve orbit and is associated with poor prognosis. On the other hand, Aspergillus and Candida are the commonest fungi found in the necrotic tumor tissue. Pathologic examination remains the hallmark in confirming the diagnosis and fungal typing.

Adolescent ; Adult ; Aged ; Antifungal Agents ; therapeutic use ; Aspergillosis ; drug therapy ; microbiology ; pathology ; Aspergillus ; isolation & purification ; Candida ; isolation & purification ; Candidiasis ; drug therapy ; microbiology ; pathology ; Carcinoma, Squamous Cell ; drug therapy ; microbiology ; pathology ; Child ; Female ; Follow-Up Studies ; Head and Neck Neoplasms ; drug therapy ; microbiology ; pathology ; Humans ; Leukemia ; drug therapy ; microbiology ; pathology ; Lymphoma, Extranodal NK-T-Cell ; drug therapy ; microbiology ; pathology ; Male ; Middle Aged ; Mycoses ; drug therapy ; microbiology ; pathology ; Retrospective Studies ; Sinusitis ; drug therapy ; microbiology ; pathology ; Young Adult ; Zygomycosis ; drug therapy ; microbiology ; pathology

OBJECTIVETo study the relationship between milk intake of 3 - 6 years old preschool children and their growth.

METHODSThe data bank of 2002 China National and Health Survey were used to analyze the relationship of milk consumption and the growth of children aged 3 - 6 in urban and rural areas.

RESULTSThe percentages of the diet with milk in urban and rural areas were 46.9% and 8.2% respectively, there was significant difference between the urban and rural areas. The averaged milk intake of preschool children were 23.8 g/d, the milk intake of urban children was significantly higher than that in the rural areas (P < 0.01). The height and weight of children with milk or its products in their diets in the group aged 4, 5 and 6 years were significantly higher than those of children without milk or its products in their diets (P < 0.05).

CONCLUSIONThere was close relationship between milk intake of children 3 - 6 years old and their growth, the volume of milk intake of preschool children should be improved in urban and rural areas.

Animals ; Body Height ; Body Weight ; Child ; Child, Preschool ; China ; Dairy Products ; Diet ; Health Surveys ; Humans ; Milk ; Rural Population ; statistics & numerical data ; Urban Population ; statistics & numerical data

AIM:To explore the effects of mammalian target of rapamycin (mTOR) double inhibitor AZD8055 on autophagy and apoptosis of human cholangiocarcinoma cell line HuCCT1. METHODS:The effect of AZD8055 on the viability of HuCCT1 cells was detected by MTT assay. Autophagosome was detected by acridine orange (AO) staining. Af-ter treated with AZD8055, the expression levels of apoptosis-related proteins Bcl-2, Bax and cleaved caspase-3 and auto-phagy marker proteins beclin 1, LC3 and p62 were determined by Western blot. Apoptotic rate was analyzed by flow cyto-metry with Annexin V-FITC/PI double staining. RESULTS:AZD8055 significantly inhibited the viability of HuCCT1 cells (P<0.05). AO staining showed that AZD8055 significantly increased orange granules in the cytoplasm. After treated with AZD8055, compared with the control group, the protein level of beclin 1 and the ratio of LC3-Ⅱ/LC3-Ⅰ were enhanced, while p62 was attenuated (P<0.05). The protein expression level of pro-apoptotic regulator Bax was down-regulated and anti-apoptotic regulator Bcl-2 was increased. The protein level of cleaved caspase-3 was reduced (P<0.05). The results of flow cytometry showed that AZD8055 inhibited cell apoptosis. CONCLUSION:AZD8055 inhibits the viability of cholangiocarcinoma cells, and the mechanism is closely related with autophagy induced by AZD8055.