AIM: To investigate the ability of Chinese cobra snake venom-metalloproteinase(MT) to induce the histamine release from human mast cells and its potential mechanisms.METHODS: MT was purified from the snake venom by using heparin agarose and Superdex75 chromatography.Mast cells were dispersed from human lung, colon and tonsil tissues after digestion with collagenase and hyaluronidase.The dispersed mast cells were then challenged with MT,stimulus and control in LP4 tubes for 15 min at 37 ℃.A glass fibre-based fluorometric assay was used to measure histamine in the supernatants of dispersed mast cells.RESULTS: MT induced a dose-dependent release of histamine from human colon,lung and tonsil mast cells.As low as 0.03(mg/L) of MT was able to stimulate significant histamine release from human colon mast cells,but a minimum of 0.3 or 30 mg/L of MT was required to stimulate a similar level of histamine release from lung or tonsil mast cells,respectively.The release of histamine from colon and lung mast cells in response to MT was maximized at 12 min following the addition of the stimulus.This was quite different from the picture of the peak histamine release from tonsil mast cells,in which histamine release was maximized at 8 min following the addition of MT.Pretreatment of cells with metabolic inhibitors and pertussis toxin reduced dramatically histamine release from human colon,lung and tonsil mast cells by MT.In exogenous Ca~(2+) and Mg~(2+) free experiments,the release of histamine induced by MT was significantly decreased.CONCLUSION: Cobra snake venom MT induces human mast cells to release histamine through a G-protein-related mechanism,which may contribute to the pathogenesis of venomous snake bite.

Objective To evaluate the potential value of IgG antibodies against recombinant PPE65 protein (rPPE65) of Mycobacterium tuberculosis in serodiagnosis of tuberculosis.Methods The gene encoding PPE65 protein of M.tuberculosis was cloned into the PET-28a vector and then expressed in Escherichia coli.The rPPE65 was purified with Ni-NTA affinity and ion exchange chromatography.After dialysis renaturation, the concentration of rPPE65 was determined using Lowry assay.ELISA was used to detect the levels of specific IgG against rPPE65 and recombinant PstS1 protein (rPstS1) in sera from 144 patients with pulmonary tuberculosis (PTB patients), 144 health controls, and 56 patients with non-tuberculosis pulmonary diseases.ROC curves were used to determine cut-off values with the results of IgG antibodies against rPPE65 and rPstS1 for 144 PTB patients and 97 controls with negative PPD skin test.The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of rPPE65 and the combination of rPPE65 and rPstS1 were counted.Results The PPE65 protein of M.tuberculosis was successfully expressed in E.coli. The purity and concentration of rPPE65 were 95% and 0.5 mg/ml, respectively.ROC analysis showed that the cut-off of ELISA using rPPE65 was 0.64.The sensitivity, specificity, PPV, NPV, and accuracy of rPPE65 were 34.7%(50/144), 93.5%(187/200), 79.4%(50/63), 66.5%(187/287), and 68.9%(237/344), respectively.The sensitivity, specificity, PPV, NPV, and accuracy of the combination of rPPE65 and rPstS1 were 59.0%, 91.0%, 82.5%, 75.5%, 77.6%, respectively.Conclusions The rPPE65 of M.tuberculosis appears to be a candidate antigen for serodiagnosis of tuberculosis.Detection of IgG antibodies against the combination of rPPE65 and rPstS1 can increase the sensitivity of serological test for tuberculosis.

Objective To explore the correlation between the distribution of uric acid (UA) level and the biochemical indicators which reflect the degree of organ lesions among hospitalized children.Methods Patients who were hospitalized to the Department of Pediatrics and received the blood UA test from June 2012 to October 2013 were included,23 217 cases in total.The biochemical analyzer-Japan's Olympus AU 2700 was used to detect blood biochemistry; and uricase-peroxidase coupling method was used to detect blood UA.Among these patients,2 099 cases whose UA level exceeded the normal level.Then the patients' gender,age,primary diagnosis and UA level were recorded; and the distribution of their UA level was described.The Chi-square or Fisher test was used to assess the incidence rate.At the same time,each blood biochemical indicators of the patients with high UA level were recorded ; and the relationships between the blood UA of the 1 650 patients with complete records and each blood biochemical indicator were analyzed with Pearson correlation analysis.Results The incidence rate of hyperuricemia among the hospitalized children was 9.04％(2 099/23 217).In particular,the incidence rate among boys and girls was 6.5t％(890/13 657) and 12.65％ (1 209/9 560) respectively (x2 =256.9,P＜0.05).The incidence rate of hyperuricemiin different diseases was as follows:in the critical illness 36.93％ (113/306),neonatal disease 20.34％ (922/4 533),urinary system diseases 12.08％ (47/389),circulatory system diseases 11.67％ (21/180),nervous system diseases 11.05％(112/1 014),digestive diseases 10.50％(190/1 810),infectious diseases 10.18％(120/1 179),blood diseases 7.58％ (55/726),endocrine system diseases 5.74％ (17/296),autoimmune diseases 4.24％ (48/1 131),respiratory diseases 3.90％ (454/11 653) respectively (x2=1423.0,P＜0.05).The incidence of hyperuricemia at younger than one month was 18.31％(929/5 075),younger than one year old was 4.22％ (359/8 501),younger than six years old was 10.68％(600/5 618),younger than 15 years old was 5.24％ (211/4 023) respectively (x2=858.5,P＜0.05).Blood UA was positively correlated to urea nitrogen,creatinine,lacticdehydrogenase,α-hydroxy-butyrate dehydrogenase,creatine kinase and creatine kinase-MB (r=0.426,0.44,0.324,0.367,0.413,0.431,P＜0.05).Blood UA was not correlated to fructosamine,blood glucose,triglycerides,total cholesterol,low-density lipoprotein and high density lipoprotein.Conclusion The incidence of hyper-uricemia among hospitalized children is high; and the incidence among children with severe diseases and newborn babies is high; followed by in children with urinary system and circulatory system diseases.The blood UA level is closely related to the blood biochemical indicators which reflect the lesions of heart and kidney.

The most common metastatic site of prostate cancer is the bone, followed by the lung, bladder, liver, and adrenal gland. We report on a rare case of pancreatic metastasis from prostate cancer. A 52-year-old patient was admitted to the hospital with epigastric pain for 20 days. PET-CT showed malignant lesions in the prostate and pancreas, and prostate and pancreas puncture biopsies were performed, respectively. The patient was diagnosed as prostate cancer with pancreatic metastasis according to the pathological findings. After undergoing androgen deprivation therapy and docetaxel chemotherapy for 6 cycles, reexamination revealed that the pancreatic metastases had disappeared.

Objective To detect the expression levels of high mobility group box chromosomal protein 1 (HMGB1) and Th17 cells transcription factors, related cytokines in peripheral blood of rheumatoid arthritis (RA) patients and analyze the relations between HMGB1 and CRP, ESR, RF in RA patients. The other aim of this study is to identify the expression level of HMGBI and the relationship between HMGB1 and Th17 in RA patients. Methods The mRNA levels of HMGB1, RORyt, interleukin (IL)-17 in the peripheral blood mononuclear cells (PBMC) were determined by quantitative real-time PCR (QRT-PCR) from 80 patients with rheumatoid arthritis,including 32 RA patients in stable phase and 48 patients in active phase, and 50 healthy volunteers. The concentration of HMGB1, IL-23, IL-17 in plasma were detected by enzyme linked immunosorbent assay (ELISA), one-way ANOVA and Spearman's correleation were adopted for statistical analysis.Results The mRNAs of HMGBI, RORyt and IL-17 in RA patients were higher than that in healthy control group (P<0.05), especially in active RA patients [ HMGB 1 (0.424±0.262) pg/ml, RORγt (0.34±0.25) pg/ml,IL-17 (1.42±0.38) pg/ml,P<0.01 ] when compared with patients with stable disease. The concentration of HMGB1, IL-23 and IL-17 in the plasma of RA patients was higher than that of the healthy control group (P< 0.05), and was positively correlated with the expression levels of HMGB1, Th 17-associated factors and the level of CRP, ESR, RF in RA patients' plasma(P<0.05). Conclusion The HMGB1 and Thl7 cells levels are higher in active RA patients than those in patients with stable disease, arid there is significant positive correlation between them. Detection of peripheral HMGB1 and Thl7 cell-specific transcription factors or related cytokines can help to understand the development and progress of rheumatoid arthritis and provide clues for new treatment targets for RA.

G heterozygote carriers.The carrying rate of deafness gene was 26‰(32/1234).In the 32 carriers,there are 5 babies showed 'refer' at the first step of hearing screening.In the 1234 babies,112 babies showed 'refer' at the first step of hearing screening.CONCLUSION Deafness gene screening can make up for the deficiencies of the universal newborn hearing screening,and should be used in this kind screening more widely.

[Abstract] Objective： ： To explore miR-103a-3p expression in the tumor tissues and the serum of breast cancer patients, and its role and mechanism in breast cancer development. Methods: Pathologically confirmed 31 cases of tumor tissues and 21 cases of para-cancerous tissues resected at Department of Oncological Surgery of the Second Affiliated Hospital of Hainan Medical University (Haikou, China) from March 1, 2017 to August 31，2017 were collected for this study; in addition, serum samples from 38 breast cancer patients and 22 healthy subjects as well as the breast cancer cell lines MCF-7 and MDA-MB-231 were used in this study. pHBLV-U6-Luc-T2A-Puro and PLL3.7 lentivirus were applied to knock down miR-103a-3p and PDK4 in MCF-7 and MDA-MB-231 cells, respectively. qPCR and Western blotting were performed to examine the mRNA and protein expressions of miR-103a-3p and PDK4 in tissues and serums of breast cancer patients as well as the in cell lines, respectively; CCK-8 assay was applied to detect the proliferation of MCF-7 and MDAMB-231 cells; Olympus AU5400 was applied to detect the glucose consumption and lactate production in indicated cell line. Results： ： miR-103a-3p was significantly decreased in tumor tissues compared with the paracancerous tissues (P<0.01). miR-103a-3p knockdown activated the glucos consumption and lactate production (all P<0.01), increased the PKD4 expression (P<0.01) in MCF-7 and MDAMD-231 cells, and promoted the proliferation of MCF-7 and MDA-MB-231 cells (P<0.01). Furthermore, knockdown of PDK4 suppressed the glucose consumption, lactate production and proliferation in MCF-7 and MDA-MB-231 cells with miR-103a-3p silencing (all P<0.01). Conclusion： ：In the breast cancer, miR-103a-3p inhibited the proliferation of breast cancer cells through down-regulation of PDK4 and PDK4-mediated aerobic glycolysis.

Objective: To compare the cytogenetic response detected by conventional banding analysis (CBA) and fluorescence in situ hybridization (FISH) and to explore the correlation between the cytogenetic and molecular response in chronic myeloid leukemia (CML) patients during tyrosine kinase inhibitor (TKI) treatment. Methods: CBA, FISH and real-time quantitative reverse transcriptase polymerase chain reaction (RQ-PCR) methods were performed to detect the cytogenetic and molecular response simultaneously in 504 bone marrow samples from 367 CML patients who received TKI treatment. Results: Among 504 samples, 344 were detected to reach complete cytogenetic response (CCyR) by CBA, while 297 samples reached CCyR by FISH which were considered to carry BCR-ABL positive cells<1%. When the results of CBA, FISH and RQ-PCR were compared in 493 samples at the same time, it showed that in 337 samples with CBA-CCyR, 273 (81.0%) reached FISH-CCyR and 289 (85.8%) were BCR-ABL^IS (International Scale, IS) ≤1% by RQ-PCR, compared to 9.0 (261/290) were BCR-ABL^IS ≤1% among 290 samples with FISH-CCyR. There was no significant difference in the median value of BCR-ABL^IS between samples in CBA-CCyR and FISH-CCyR (0.21% vs 0.13%, z=-1.875, P=0.061) . Furthermore, when the samples were divided into three groups according to BCR-ABL positive cells (0,>0~<1%, 1%~5%) by FISH, the statistical difference was observed, the proportion of samples with BCR-ABL^IS ≤1% in the three groups were 94.1%, 57.6% and 27.7% respectively (χ²=43.499, P<0.001; χ²=9.734, P=0.003) , while the median value of BCR-ABL^IS were 0.10%, 0.64% and 1.80% respectively (z=-5.864, P<0.001; z=-4.787, P<0.001) . Conclusion FISH results were in good concordance with CBA in identify samples in CCyR, FISH was more sensitive and had better correlation with RQ-PCR results than CBA, but how to define FISH-CCyR need further study.

Objective: To analyze the characteristics of myeloid neoplasms with t (3;21) (q26;q22) . Methods: Clinical data of patients with t (3; 21) (q26; q22) , diagnosed as hematologic malignancies in Peking University people's hospital from January 2011 to March 2018, were collected retrospectively. 19 patients in our hospital and forty-eight patients bearing t (3;21) (q26;q22) with detailed survival data reported in literature were summarized. Kaplan- Meier method was used for survival analysis. Results: Among 19 patients, including 15 males and 4 females with a median age of 36 years (22-68 years) , 4 cases was diagnosed as de novo acute myeloid leukemia (AML) , 4 as myelodysplastic syndromes (MDS) , 3 as MDS-AML and 8 as chronic myelogenous leukemia (CML) in myeloid blast transformation. All of the 19 patients were detected to have t (3;21) (q26;q22) by G-banding technique and 13 carried additional cytogenetic aberrations. 9 of the 19 patients were detected for positive AML1-MDS1 fusion genes. In the 9 patients with detailed follow-up data, 6 patients received chemotherapy and only 2 achieved complete remission (CR) while 4 with no response. During the follow-up period, 8 patients died and the median overall survival (OS) was 6 months (4.5 to 22 months) . Survival analysis of the present 9 patients together with the literature data showed that the prognosis was poor and the median OS was 7 months. In particular, AML/t-AML had the worst prognosis. Hematopoietic stem cell transplantation (HSCT) could significantly improve survival, the median OS in HSCT group and non-HSCT group were 20.9 and 4.7 months respectively (P<0.001) . Conclusions t (3; 21) (q26; q22) is a rare recurrent chromosomal abnormality which is detected mainly in myeloid neoplasm and confer to poor clinical prognosis. HSCT should be recommended to improve the outcomes.

OBJECTIVETo report on a case of therapy-related acute monocytic leukemia(t-AML) with t(11;17) (q23;q21)/MLL-AF17q after successful treatment for acute promyelocytic leukemia(APL) with t(15;17) (q22;q21)/PML-RARα.

METHODSA MICM method (bone marrow morphology(M), immunophenotype(I), cytogenetics(C), and molecular biology(M)) was used for the diagnosis and classification of the disease at the time of onset and transformation.

RESULTSThe patient was initially identified with typical morphology and immunophenotype of APL. She has carried t(15;17)(q22;q21) and PML-RARα fusion gene but was without t(11;17)(q23;q21) or MLL gene abnormalities. After 13 months of successful treatment, she has transformed to AML with typical morphology and immunophenotype. t(11;17)(q23;q21) and MLL-AF17q fusion gene were detected in her bone marrow sample, while no PLZF-RARα fusion gene was detected by real-time quantitative reverse-transcription PCR(RQ-PCR) and fluorescence in situ hybridization(FISH).

CONCLUSIONt-AML is a serious complication after successful treatment of APL. t(11;17)(q23;q21) is not specific for the diagnosis of variant APL and can also be detected in t-AML. RQ-PCR and FISH are essential for the diagnosis of such patients.

Chromosomes, Human, Pair 11 ; Chromosomes, Human, Pair 15 ; Chromosomes, Human, Pair 17 ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Monocytic, Acute ; genetics ; Leukemia, Promyelocytic, Acute ; genetics ; Middle Aged ; Neoplasms, Second Primary ; genetics