Objective To observe the pathology in temporal lobe cortexes and hippocampus in protopathic intractable temporal lobe epileptics.Methods The cortexes of spike foci in temporal lobe and hippocampus were obtained from 9 cases with protopathic intractable temporal lobe epilepsy who accepted operation.The samples were observed under the transmission electron microscope.Results The ultrastructure changes in spike focus of temporal lobe cortexes are similar to those in hippocampus.It is common that neurons were pycnotic and decreased.Astrocytes were hydropic and degenerative.Gliosis were found in some cases.The number of synapses increased or decreased in different cases and positions.Blood-brain barriers were destroyed because foot processes of astrocytes around capillaries were edematous.Conclusion Neuronal loss,gliosis and synaptic reorganization which occur in epileptic hippocampus and maybe also in epileptic temporal lobe cortexes destroy the balance between excitatory and inhibitory neurotransmission.The abnormalities probably associate with protopathic intractable temporal lobe epileptic seizures.

Objective To establish and evaluate the new method of 8-color flow cytometry (8c-FCM) with two tube detecting bone marrow minimal residual disease (MRD) in B lineage acute lymphoblastic leukemia (B-ALL).MethodsThe MRD cells were analyzed by using two combinations of 8c-FCM antibody panels,gating with CD19/Side scatter(SSC),CD45/CD10 and CD34(or cTdT).Nalm-6 cell of B-ALL was mixed into normal marrow cells,with proportion of 10.00％,1.00％,0.10％,0.01％and 0.005％,and recovery test and reproducibility test were carried with 8c-FCM established to value its accuracy and precision of.Fluorescence intensity was detected on different time points after marked Nalm-6 by antibodies to evaluate the fluorescent stability of the antibodies.The immunophenotyping was analyzed in 39 bone marrow specimens,including 9 cases of normal control,9 cases of B-ALL primary or recurrent,21 cases of complete remission (CR) after chemotherapy or bone marrow transplantation ( BMT),to evaluate the detection of normal B lymphocyte lineage,leukemia cells of B-ALL,MRD cells of B-ALL using the 8c-FCM and compare it with 4c-FCM in being.ResultsThe method of two tube 8c-FCM with main antibodies of CD19,CD45 and CD10 to detect MRD of B-ALL was founded; CV ＜ 2.5％,average recovery rate was 95.81％,when the actual percent of Nalm-6 mixed into normal bone marrow≥0.10％,and the percent of Nalm-6 detected and actual was linear dependent ( r =0.99,P ＜ 0.05 ) ranged from 10.00％ - 0.01 ％,when 106 cells were acquired ; the sensitivity of the method established could reach 0.01％.The fluorescent intensity decreased along with the time after Nalm-6 cell marked,but less than 10％ in 24 hours.Using the antibody combinations and analyze strategy,the immunophenotye of B lymphocyte in normal bone marrow presented four sequential stages:Stage Ⅰ CD45low/CD10stro/CD20 -/CD38stro/CD34 + or cTdT +,Stage ⅡCD45 +/CD10 + / CD20 -/CD38stro/CD34+ or cTdT +,Stage Ⅲ CD45 +/CD10 +/CD20 -/CD38stro/CD34 -or cTdT-,Stage Ⅳ CD45stro/CD10 -/CD20 +/CD38low/CD34or cTdT.Antigen expressions of leukeamic cells of B-ALL primary or recurrent were different compared with control team; there were 5 cases with MRD positive in CR team,and the main antigen expression was consistent with the results from 4c-FCM.The range of the percent of MRD cells was 0.02％ - 5.42％ of the 5 cases of MRD positive.ConclusionsThe new method of two tube 8c-FCM established shows good reproducibility and high accuracy,and can identify normal B lymphocyte populations in bone marrow and regenerated B-precursors in CR cases with MRD cells;compared with 4c-FCM,the new method of two tube 8c-FCM with the fewer specimen is faster and efficient to diagnose MRD of B-ALL.

Cystectomy ; Female ; Humans ; Intrauterine Devices

Animals ; Aorta ; pathology ; Atherosclerosis ; blood ; etiology ; metabolism ; Chemokine CCL2 ; metabolism ; Cholesterol ; blood ; Cholesterol, Dietary ; administration & dosage ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Dehydroepiandrosterone ; pharmacology ; Diet, Atherogenic ; Immunohistochemistry ; Rabbits ; Random Allocation ; Triglycerides ; blood ; Vascular Cell Adhesion Molecule-1 ; metabolism

Objective To explore the values of potential clinical application ofsingle tube/ten colorsflow cytometry for leukocyte differential count in peripheral blood.Methods Utilizing multiple monoclonal antibody combinations and the vavious logical gating strategies,the single tube/12 antibodies with no-wash method for the leukocyte differential count in peripheral blood were determined by using 10 colors flow cytometry.Leukocyte differentials of 142 peripheral blood samples were determined by both Beckman-Coulter LH750 hematology analyzer and 10 colors flow cytometry.The results were then compared to standard microscopic examination as a reference method.The clinical diagnostic efficiency ofsingle tube/10 colorsflow cytometry was calculated.The correlations between standard microscopic cytology,single tube/10 colorsflow cytometry and the hematology analyzer were determined.In addition,the clinical diagnosis efficiency for blast counts ofsingle tube/10 colorswere compared to the results determined by BD FACS Calibur flow cytometer.Results The leukocyte differentials were correlated well between the single tube/10 colorsflow cytometry and standard microscopic cytology(r>0.700,P<0.01) except for basophils.The correlations with neutrophilic granulocytes,lymphocytes,immature granulocytes and blasts were superior(r=0.972,0.951,0.801,0.912,respectively,P<0.01).When 1% was selected as the cut-off point for immature granulocytes determined by standard microscopic cytology,the sensitivity and the specificity ofsingle tube/10 colorsflow cytometry were 92%(57/62) and 79% (63/80),respectively.When 0.5% was selected as the cut-off point for blasts detected by standard microscopic cytology,the sensitivity and the specificity were 99% (67/68) and 92% (68/74).Using the immunophenotyping results from BD FACS Calibur as a standard,the sensitivity for detecting blasts bysingle tube/10 colOrsflow cytometry was 100% (40/40),the specificity was 91% (10/11),the positive predictive value was 98% (40/41),the negative predictive value was 100% (10/10) and the accuracy was 98% (50/51).Conclusions Thesingle tube/10 colorsflow cytometry has a excellent correlation with the standard microscopic cytology when applied on leukocyte differential count in peripheral blood.It may potentially use as a subsequent method for verification of abnormal results of complete blood cell count in the future.

OBJECTIVETo investigate the effects of total flavonoids of Litsea Coreana (TFLC) on the gap junction (GJ) intercellular communication in TM3 testicular Leydig cells and whether TFLC can reduce the cytotoxicity of oxaliplatin (OHP) in vitro.

METHODSWe detected the effect of TFLC on the dye spread of the in vitro cultured TM3 cells by parachute assay, observed changes in the expression of connexin 43 (Cx43) total protein in the TFLC-treated TM3 cells by Western blot, and determined the effects of TFLC on the expression of Cx43 on the membrane of the TM3 cells by immunofluorescence assay and on the cytotoxicity of OHP by MTT assay.

RESULTSTFLC obviously enhanced the GJ function with the increasing of the TFLC concentration in the TM3 cells. Western blot and immunofluorescence assay confirmed that TFLC significantly enhanced the expression of Cx43 total protein and Cx43 expression on the membrane of the TM3 cells. MTT assay showed that at a high cell density (confluent with GJ formation), 20 microg/ml TFLC enhanced the GJ function of the TM3 cells and reduced the cytotoxicity of OHP (P < 0.05), while at a low density (preconfluent with no GJ formation), TFLC exhibited no effect on the cytotoxicity of OHP (P > 0.05).

CONCLUSIONTFLC increases the Cx43 expression and GJ function in normal TM3 Leydig cells, and the enhancement of GJ function reduces the cytotoxicity of OHP.

Antineoplastic Agents ; toxicity ; Cell Communication ; drug effects ; physiology ; Cell Count ; Connexin 43 ; metabolism ; Flavonoids ; pharmacology ; Gap Junctions ; drug effects ; Humans ; In Vitro Techniques ; Leydig Cells ; drug effects ; ultrastructure ; Litsea ; chemistry ; Male ; Organoplatinum Compounds ; antagonists & inhibitors ; toxicity ; Proteins ; metabolism

OBJECTIVETo retrospectively assess serious systemic adverse effects of standardized dust-mite vaccine in children with asthma.

METHODSMedical records of 704 children (5-17 years in age) with asthma between January, 2005 and December, 2011 were reviewed. Serious systemic adverse events following treatment with a standardized dust-mite vaccine in these children were analyzed.

RESULTSA total of 336 systemic adverse reactions were observed in 17.0% (120/704) of the patients analyzed of these adverse reactions, 18 (5.4%) were serious (level 3), 318 (94.6%) were not serious (below level 3), and no single case of anaphylactic shock (level 4) was recorded. Systemic adverse events occurred most frequently in the 5 to 11-year age group and in the summer season (from June to August). In the 18 severe cases, the peak expiratory flow (PEF) dropped by 20% immediately after the vaccine injection, and other major clinical symptoms included cough, wheezing and urticaria. All children with serious systemic adverse effects were given inhaled hormone and atomized short-acting beta agonists, oral antihistamines, intravenous dexamethasone and/or intramuscular adrenaline. After these treatments, the clinical symptoms were significantly relieved.

CONCLUSIONSThe rate of serious systemic adverse events following allergen-specific immunotherapy is relatively low in children with allergic asthma. Conventional medications are effective in managing these immunotherapy-associated adverse events.

Adolescent ; Animals ; Asthma ; physiopathology ; therapy ; Child ; Child, Preschool ; Desensitization, Immunologic ; adverse effects ; Female ; Humans ; Male ; Peak Expiratory Flow Rate ; Pyroglyphidae ; immunology ; Retrospective Studies ; Vaccines ; adverse effects

OBJECTIVE: To determine the expressions of Notch1, Jagged1 and vascular endothelial growth factor (VEGF) in human non-small cell lung cancer (NSCLC) and to explore its clinical pathological significance. METHODS: Immunohistochemical SP method was used to detect the expressions of Notch1, Jagged1 and VEGF in 65 patients with NSCLC and 15 normal epithelial tissues of the lung, and the relationship between them and clinic-pathological parameters were analyzed. RESULTS: The positive rates of Notch1, Jagged1 and VEGF in NSCLC were 81.5%, 83.1% and 93.8%, respectively, higher than those in normal epithelial tissues of the lung (P<0.05). The positive expression levels of Notch1 and VEGF were closely associated with the tumor stage and the lymph node metastasis (P<0.05). The positive expression levels of Jagged1 was positively correlated with the pathological type and lymph node metastasis. There was a positive correlation between Notch1, Jagged1 and VEGF. CONCLUSION Notch1, Jagged1 and VEGF protein may play an important role in the pathway of carcinogenesis and progression of NSCLC. The up-regulation of Notch1, Jagged1 and VEGF protein expression probably predict NSCLC carrying relatively strong permeation and metastasis.
Adult ; Aged ; Calcium-Binding Proteins ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Jagged-1 Protein ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Membrane Proteins ; metabolism ; Middle Aged ; Neoplasm Staging ; Receptor, Notch1 ; metabolism ; Serrate-Jagged Proteins ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the effect of Hedgehog (HH) pathway on proliferation and in vitro tumorigenicity of gastric cancer cell lines.

METHODSThe expression of SHH, PTCH, SMO, SUFU and GLI1 in seven cell lines were tested by RT-PCR. siRNA targeting GLI1 mRNA was transfected into MKN28 cells. Cell proliferation and in vitro tumorigenicity were examined by CCK8 and soft agar colony formation test.

RESULTSSHH in six gastric cancer cell lines was up-regulated. Expression of PTCH in KATOIII cell lines and expression of SUFU in MKN28 and KATOIII were reduced. GLI1 siRNA significantly inhibited the expression of GLI1 in MKN28 cell line. Growth rate and colony formation rate of MKN28 cells treated with GLI1 siRNA were significantly lower than those of control cells (all P <0.001).

CONCLUSIONSHH signaling pathway is widely activated in gastric cancer cell lines. The activation of HH signaling pathway promotes the growth of MKN28 cells.

Cell Line, Tumor ; Cell Proliferation ; Gastric Mucosa ; cytology ; Hedgehog Proteins ; metabolism ; Humans ; Oncogene Proteins ; metabolism ; RNA, Small Interfering ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology ; Trans-Activators ; metabolism ; Zinc Finger Protein GLI1

Animals ; Carcinoma, Hepatocellular ; genetics ; metabolism ; virology ; Genes, p53 ; Hepatitis B ; genetics ; metabolism ; virology ; Hepatitis B virus ; Humans ; Inhibitor of Growth Protein 1 ; Intracellular Signaling Peptides and Proteins ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; virology ; Loss of Heterozygosity ; Nuclear Proteins ; metabolism ; Point Mutation ; Signal Transduction ; Trans-Activators ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Tumor Suppressor Proteins ; metabolism