Allergic reactions caused by traditional Chinese medicine injections (TCMIs) become a greatest concern in the clinic application safety. The integral animal evaluation method commonly used in the preclinical evaluation for allergic reactions of TCMIs was not sensitive, specific, quick and objective in observation indexes. Therefore, more researchers have paid attention to the in vitro test method for evaluating allergic reactions induced by TCMIs. Currently, the methods for evaluating allergic reactions induced by TCMIs are mainly targeted at type I allergic reaction and anaphylactic reaction, with only a few in vitro methods for evaluating type II allergic reaction. In this paper, researchers summarized relevant literatures published about evaluation methods for allergic reactions induced by TCMIs in recent years.
Animals ; Complement Activation ; Drug Hypersensitivity ; diagnosis ; etiology ; Humans ; Injections ; Medicine, Chinese Traditional ; adverse effects

Objective To evaluate the diagnostic value and safety of endobronchial ultrasoundguided transbronchial needle aspiration (EBUS-TBNA) in elderly patients with bronchogenic carcinoma.Methods From August,2013 and July,2014,31 patients aged 65 years and over with space-occupying mass and enlarged mediastinal lymph nodes detected by CT or positron emission tomography-computed tomography (PET-CT) underwent EBUS-TBNA.Rapid onsite cytology evaluation was not performed.Results There were 26 males and 5 females in this study,aged 65-77 years (70.1 years old on average).In 31 patients,70 samples were obtained from lymph nodes (LNs) and 4 samples were obtained from intrapulmonary lesions.29 cases were diagnosed as lung cancer,and 2 cases had false-negative diagnoses.The sensitivity and specificity rates of EBUS-guided TBNA method were 93.6％ and 100.0％,respectively.No major complications were observed in this series.Conclusions EBUS-TBNA is an safe and effective method in diagnosing bronchogenic carcinoma in elderly patients.

Adolescent ; Child ; Child, Preschool ; Chromosomes, Human, Pair 6 ; Female ; Humans ; Infant ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics

Objective To investigate the effect of recombinant human bone morphogenetic protein 2 (rhBMP-2) on the osteogenic activities of human osteosarcoma cell line SaOS-2. Methods SaOS-2 cells were exposed to rhBMP-2 for 12,24,48 h at 0(control) ,2,20,200 μg/L, respectively. The mRNA expression of alkaline phosphatase(ALP) and bone gla(BCP) were detected by real time polymerase chain reaction. Results The mRNA expression of ALP and BGP of SaOS-2 cells increased gradually with rhBMP-2. The mRNA expression of ALP of the 20 μg/L group exposed for 48 h(1.60 ± 0.64), and the 200 μg/L group exposed for 12,48 h(1.70 ± 0.41, 1.80±0.19) were significantly higher than those of control (12 h: 0.80±0.25, 48 h: 0.74±0.21, allP<0.05). The mRNA expression of BGP of the 2 μg/L group exposed for 24 h(1.67 ± 0.33), the 20 μg/L group exposed for 12,24 h(2.42 ± 0.13,1.82 ± 0.14) and the 200 μg/L group exposed for 12,24 h(1.46 ± 0.11,1.24 ± 0.07) were significantly higher than those of control( 12 h: 1.01 ± 0.14, 24 h: 0.84 ± 0.12, all P< 0.05). Conclusions rhBMP-2 can promote the mRNA expression of ALP and BGP of SaOS-2 cells. They have a dose-response relationship, but represent a different dose-response effect.

Objective To observe the effects of recombinant human parathyroid hormone 1 to 34(referred to as hPTH) on the expression level of alkaline phosphatase(ALP) and bone gla protein(BCP) in human osteosarcoma cell line SaOS-2(referred to as SaOS-2 cells). Methods SaOS-2 cells were subcultured and treated with 1, 10 and 100 nmol/L hPTH for 12, 24 and 48 h. Total cellular RNA was extracted, cDNA was synthesized by reverse doses of hPTH, different duration of action, and their interaction on the expression level of ALP mRNA of SaOS-2 cells was significantly different(F = 29.32, 2.92, 7.64, all P < 0.05). The expression level of ALP mRNA(0.78 ± 0.43, 0.71 ± 0.05, 0.75 ± 0.19, 0.76 ± 0.14) of SaOS-2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h(1.01 ± 0.16, 1.37 ± 0.38, 1.49 ± 0.16, 2.52 ± 0.70, all P< 0.05) and 24 h (1.80 ± 0.47, 1.30 ± 0.36, 1.27 ± 0.17, 1.17 ± 0.11, all P< 0.05). The expression level of ALP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 hours was higher than that of the control(P < 0.05); the expression level of ALP mRNA of SaOS-2 cells after treatment with 1, 10 and 100 nmol/L hPTH for 24 h interaction on the expression level of BGP mRNA of SaOS-2 were significantly different (F = 8.26, 10.33, 5.51, all P< 0.05). The expression level of BGP mRNA(1.17 ± 0.28, 0.98 ± 0.08, 0.92 ± 0.17 and 0.84 ± 0.59) of SaOS2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h( 1.01 ± 0.14, 1.21 ± 0.18, 1.34 ± 0.30, 1.68 ± 0.62, all P< 0.05), and 24 h(1.71 ± 0.35, 1.41 ± 0.47, 1.28 ± 0.31 and 1.01 ± 0.18, all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 h was higher than that of those groups treated with 0 and 1 nmol/L hPTH(all P< 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 10 and 100 nmol/L hPTH for 24 h and 48 h was lower than those of the control(all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 24 hours was lower than that the group treated with 1 nmol/L hPTH(P < 0.05). Conclusions In vitro, hPTH significantly enhances osteogenic activities of human osteoblast in a short time, however, with prolonged stimulation time, osteogenic activity can show a downward trend.

OBJECTIVEThis study analyzed the clinical data of newly diagnosed childhood leukemia from various hospitals in the cities or counties of Hunan Province between 2002 and 2005 in order to provide references for further epidemiologic survey of childhood leukemia.

METHODSThe clinical data of children with newly diagnosed leukemia from hospitals of various cities or counties of Hunan Province between 2002 and 2005 were collected and reviewed.

RESULTSThere were 803 children with leukemia during 2002-2005. Acute lymphoid leukemia was most commonly seen (597/803, 74.35%), followed by acute non-lymphoid leukemia (192/803, 23.91%) and chronic myelocytic leukemia (14/803, 1.74%). There were no significant differences in the clinical type and the prevalence of leukemia between males and females. The prevalence of newly diagnosed childhood leukemia in the urban area was noticeably higher than that in the rural area (2.02/10(5) vs 1.50/10(5), P < 0.05). 41.79% of children with newly diagnosed leukemia from the urban area received treatments but only 22.80% of patients from the rural area received treatments (P < 0.05).

CONCLUSIONSThis study of patients-based hospitals showed some features of the morbidity of childhood leukemia in Hunan Province. It provides references for further epidemiologic investigation of this disease in Hunan Province.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Incidence ; Infant ; Infant, Newborn ; Leukemia ; diagnosis ; epidemiology ; therapy ; Male ; Time Factors

OBJECTIVETo examine the long-term outcomes of total colonic aganglionosis (TCA) and to evaluate their nutritional status.

METHODSEleven pediatric patients treated for TCA between January 1999 and December 2010 were included in the study and followed up. Physical measurements including height, weight and laboratory tests were assessed. Anorectal functions were evaluated with Kelly score and quality of life(QOL) using questionnaire.

RESULTSThe length of follow-up ranged from 8 to 147 months. The children had satisfactory anorectal function (Kelly score, 5-6). One child had a Kelly score of 3. The children who were followed up less than 48 months had significant higher Kelly scores compared with those with more than 48 months follow-up(P<0.05). QOL was good in nine patients (QOL score, 9-10) and moderate (score, 7-8) in 2 patients. Weight-for-age was normal in 2 patients, mild malnutrition in 6 patients, and moderate malnutrition in 3 patients. Height-for-age was normal in 6 patients, mild malnutrition in 3 patients, and moderate malnutrition in 2 patients. The serum albumin was(49.0±2.7) g/L in children with well-educated parents, significantly higher than those with poorly-educated parents(44.3±1.9) g/L(P<0.05).

CONCLUSIONSLong-term outcomes of children with TCA are satisfactory with good anorectal function and quality of life. Low body weight is more common than low height. Children with well-educated parents have better nutrition status.

Follow-Up Studies ; Hirschsprung Disease ; surgery ; Humans ; Infant ; Male ; Nutritional Status ; Treatment Outcome

This study is to investigate the effect of compound B2 on the damage of PC12 cells induced by serum deprivation and to explore its related mechanisms. The binding characteristics of B2 to rat striatum adenosine A2A receptor was studied by radioligand 3H-MSX-2 binding assay. Cell viability was detected by MTT assay. ROS formation was measured after DCFDA fluorescent staining. B2 has affinity to rat adenosine A2A receptor (K1 = 0.37 micromol x L(-1)). B2 remarkably increased PC12 cell survival rate in serum deprivation-induced PC12 cells. The percentage of serum deprivation-induced death of PC12 was 49.6%, and the treatment of B2 (0.1-100 micromol x L(-1)) increased the cell viability to 63.3%, 74.9%, 86.3% and 88.1%, respectively. Adenosine A2A receptor antagonist SCH 58261 could significantly block the protective effect of B2. The cell viability with 0.1 micromol x L(-1) SCH 58261 decreased by 16.1%, 24.0% and 19.8%, in the presence of B2 (0.1-10 micromol x L(-1)). Serum deprivation-induced ROS formation was 3.5 times more than that of control group, and treatment with B2 significantly and dose-dependently inhibited ROS over-formation. The protective effect of B2 may be related with adenosine A2A receptor. Decrease of serum-deprivation induced ROS formation may also be one of the mechanisms.
Adenosine ; analogs & derivatives ; metabolism ; pharmacology ; Adenosine A2 Receptor Antagonists ; pharmacology ; Animals ; Cell Survival ; drug effects ; Corpus Striatum ; metabolism ; Culture Media, Serum-Free ; Female ; Male ; PC12 Cells ; Pyrimidines ; pharmacology ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Receptor, Adenosine A2A ; metabolism ; Triazoles ; pharmacology

OBJECTIVETo analyze the expression of genes in the Slit/Robo signaling pathway, and the methylation status of their promoters in hepatocellular carcinoma (HCC) cell lines.

METHODSGenomic DNA and total RNA were isolated from 9 HCC cell lines of different metastatic ability (Hep3B, HepG2, PLC/PRF/5, SMMC-7721, BEL-7402, MHCC97-H, MHCC97-L, LM3, LM6) and a control cell line L-02. The expression profiles of Slit1, Slit2, Slit3, Robo1, and Robo3 were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The methylation status of the promoters was detected by methylation specific polymerase chain reaction (MSP).

RESULTSThe promoters of Slit1, Slit2 and Slit3 genes were almost methylated in all the HCC cell lines. The Slit1 and Slit3 RNAs were not detected in most of the cell lines. Furthermore, the mRNA Slit2 was decreased gradually as the metastatic potential of the cell lines increased. As the candidate ligand of the Slit2 gene, Robo1 was frequently methylated in HCC cell lines whereas its mRNA was detected in all of these cells except SMMC-7721, BEL-7402 and L-02. Robo3 was unmethylated in HCC cell lines while its mRNA was not detected in these HCC cell lines.

CONCLUSIONThe hypermethylation status of Slit/Robo signaling pathway related genes is a universal event in the HCC. The hypermethylation status of Slit1, Slit2, Slit3 genes associated with the loss of expression or reduced expression. Those data suggest that Slit/Robo pathway may play a significant role in the progress or metastasis of HCC.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; CpG Islands ; genetics ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Membrane Proteins ; genetics ; metabolism ; Nerve Tissue Proteins ; genetics ; metabolism ; Promoter Regions, Genetic ; Receptors, Immunologic ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction

OBJECTIVETo profile the methylation alterations of CpG islands in hetpatocellular carcinoma cell lines.

METHODSA global analysis of DNA methylation using the Human CpG-island 12K Array (HCGI12K) from Canada University Health Network was performed on nine human hepatocellular carcinoma (HCC) cell lines (Hep3B, HepG2, PLC/RPF/5/RPF/5, SMMC-7721, BEL-7402, MHCC97-H, MHCC97-L, HCCLM3, HCCLM6) and a control cell line Chang's liver. Metastatic potential related alterations were also screened in MHCC97 series cell lines (MHCC97-H, MHCC97-L, HCCLM3, HCCLM6), using MHCC97-L, a cell line with low metastatic potential, as control. To screen the key genes which are hypermethylation or hypomethylation in the HCC cell lines compared with the normal liver cell line by normalization processing and cluster analysis of microarray data. Two randomly selected genes was analyzed by methylation specific PCR to verify the chip results.

RESULTSBy a standard of methylation alteration ratio > or = 2 or < or = 0.5, fifty-eight CpG island cloning sites and sixty-six upstream or downstream tumor-related genes were identified. The genes were oncogenes, tumour suppressor genes and their ligand genes, apoptosis-related genes, cell proliferation and differentiation genes, cell cycle-related gene and cell signaling pathway key genes such as Wnt, ras, and FGF pathway-related genes. The methylation specific PCR results were consistent with those obtained by chips.

CONCLUSIONThe results of this study demonstrate that there are a series of CpG island methylation alterations in HCC cell lines. The expression of many oncogenes, tumor suppressor genes and other key genes may be up- or down-regulated, respectively, because of their CpG island hypomethylation or hypermethylation accordingly. It may provide a basis for screening HCC biological markers by CpG island methylation profilling.

Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line ; Cell Line, Tumor ; CpG Islands ; genetics ; DNA Methylation ; Gene Expression Profiling ; methods ; Genes, Neoplasm ; Humans ; Liver ; cytology ; Liver Neoplasms ; genetics ; pathology ; Oligonucleotide Array Sequence Analysis