Cells, Cultured ; Chemokine CCL4 ; Chemotaxis, Leukocyte ; drug effects ; Endothelial Cells ; cytology ; metabolism ; Homocysteine ; pharmacology ; Humans ; Macrophage Inflammatory Proteins ; biosynthesis ; genetics ; Monocytes ; physiology ; RNA, Messenger ; biosynthesis ; genetics ; Umbilical Veins ; cytology

The method of monoclonal antibody-based immunoassay has a great importance in the study of quality control of traditional Chinese medicine (TCM) and detection of trace components in vivo animals. Synthesis of small molecule artificial antigen is the prerequisite for the establishment of this method. In present study, catalpol-BSA was synthesized by sodium periodate oxidation method. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry ( MALDI-TOF-MS) and molecular exclusion chromatography showed that catalpol was successfully conjugated with BSA. The mice could specifically produce anti-catalpol antibodies with titer up to 1:8000. The artificial antigen of catalpol was successfully synthesized.
Animals ; Antibodies ; immunology ; Antigens ; chemistry ; immunology ; Immunoassay ; Iridoid Glucosides ; chemistry ; immunology ; Male ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred BALB C ; Serum Albumin, Bovine ; chemistry ; immunology

OBJECTIVETo investigate the clinical characteristics of children with critical hand-foot-mouth disease (HFMD) who were treated with mechanical ventilation and to explore the risk factors for poor prognosis.

METHODSThe clinical data of 63 children with critical HFMD who were admitted to the pediatric intensive care unit between April 2012 and September 2013 and needed mechanical ventilation were retrospectively analyzed.

RESULTSAmong the 63 children, 43 were boys and 20 were girls, and their mean age was 25 ± 18 months, with 81% under 3 years old. The four death cases were all under three years old. Compared with the cured cases, the death cases had a significantly lower mean age (8 ± 3 months vs 25 ± 18 months; P<0.05). Poor peripheral circulation above the elbow or knee joint, pulmonary edema involving at least two thirds of the lung field, and pulmonary hemorrhage were all closely related to death (P<0.01). The death cases and cured cases had significantly different peripheral white blood cell counts, blood lactic acid, and blood glucose (24 ± 11× 10⁹/L vs 12 ± 5×10⁹/L; 6.6 ± 1.8 mmol/L vs 3.6 ± 1.7 mmol/L; 16.4 ± 2.5 mmol/L vs 10.0 ± 3.0 mmol/L). The cases with critical illness score <90 had a significantly higher death risk (P<0.01).

CONCLUSIONSChildren with critical HFMD are mainly under 3 years old. The children face extremely high risk of death when they suffer from poor peripheral circulation above the elbow or knee joint, pulmonary edema involving at least two thirds of the lung field, and pulmonary hemorrhage. Significant increases in peripheral white blood cell counts, blood lactic acid, and blood glucose are risk factors for poor prognosis. Critical illness score is also related to poor prognosis.

Blood Glucose ; analysis ; Child ; Child, Preschool ; Female ; Hand, Foot and Mouth Disease ; blood ; mortality ; therapy ; Humans ; Infant ; Lactic Acid ; blood ; Leukocyte Count ; Male ; Prognosis ; Respiration, Artificial ; Retrospective Studies

OBJECTIVETo investigate the relationship between vascular endothelial growth factor D (VEGF-D) and metastasis of lymphatic vessel in gastric carcinoma.

METHODSThe human VEGF-D cDNA was amplified from total RNA isolated from human normal gastric tissue, then it was inserted into T-A clone plasmid and subcloned into pEGFP eukaryotic expression vector. After the full-length sequence expected was confirmed by enzymatic digestion and sequencing,the human gastric carcinoma cell line SGC7901, which expressed a low level of VEGF-D, was transfected with the pEGFP/VEGF-D expression vector. Stable SGC7901 clones with high expression of VEGF-D were selected in vitro with G418, which were then combined and subcutaneously injected into nude mice to observe the density and morphology of lymphatic vessel. The outcomes were later compared with those of SGC7901 cells transfected with null vector(pEGFP) by immunostaining with a specific antibody LYVE-1.

RESULTSThe average weight of tumors in the pEGFP group (1.13+/-0.40) g at day 35, was significantly lower than that in the pEGFP/VEGF-D group (2.24+/-0.82)g (P<0.05). The lymphatic vessel density (LVD) in the pEGFP group (2.89+/-1.32) was significantly lower than that in the pEGFP/VEGF-D group (5.74+/-1.30)(P<0.01). There were dilated functional lymphatic vessels around the tumor margin.

CONCLUSIONVEGF-D may promote the growth and metastasis of tumor in gastric carcinoma by increasing the growth of lymphatic vessels.

Animals ; Cell Line, Tumor ; Humans ; Lymphatic Metastasis ; pathology ; Lymphatic Vessels ; pathology ; Mice ; Mice, Nude ; Stomach Neoplasms ; pathology ; Transfection ; Vascular Endothelial Growth Factor D ; genetics

Objective To evaluate the performance of diluted thrombin time (dTT) assay for detecting Dabigatran levels and observe whether this assay may meet the requirements of clinical laboratory.Methods According to EP15-A2,EP6-A,EP7-A and C-24 documents of the Clinical and Laboratory Standards Institute (CLSI),the precision,trueness,analytical measurement range,carryover rate and anti-biological interference of dTT assay were evaluated and the stability of specimen for dTT assay was observed.Results Both the within-day and between-day coefficient of variation (CV) of dTT assay for detecting Dabigatran levels were consistent with manufacturer's stated CV.Compared with target values of Dabigatran,the relative bias of 3 levels of proficiency test materials from College of American Pathologists (CAP) were less than 10％.The results meet linear verification when Dabigatran concentration was between 30.92 and 249.13 ng/mL.The carryover rate was-0.84％.There was no interference for Dabigatran levels by dTT assay for detecting Dabigatran when Hb≤3 g/L,triglyceride≤873 mg/dL,heparin≤2.2 IU/mL and FDP≤29 mg/L.The results of stability showed that plasma specimens for dTT could not be stored at room temperature more than 4 hours,at 4 ℃ more than 4 days,at-20 ℃ exceed 1 month,while at-80℃ the plasma specimens could be stored at least 6 months for dTT assay.Conclusion The precision,trueness,analytical measurement range,carryover rate,anti-biological interference of dTT assay may meet the requirement of clinical laboratory.The stability of the specimen can fulfill the clinical requirements.

A retrospective analysis was performed for the clinical data of four children with Epstein-Barr virus (EBV)-related acute liver failure. There were two boys and two girls with a median age of 10 months (range 8.5-44 months). Of the four children, three were diagnosed with infectious mononucleosis (IM), among whom two met the diagnostic criteria of hemophagocytic lymphohistiocytosis (HLH), and one was diagnosed with past EBV infection. All the children had positive EBV DNA in blood and all had pyrexia, hepatomegaly, and jaundice on admission. Three children had the symptom of splenomegaly, ascites, or vomiting. Two children had enlargement of cervical lymph nodes, skin rash, or pleural effusion. One child had gastrointestinal bleeding or stage 2 hepatic encephalopathy. All the children had an abnormal lymphocyte count of <10%, and only one child had leukocytosis and thrombocytopenia. Among the four children, alanine aminotransferase level increased by 10-100 times; total bilirubin level increased by 3-5 times; lactate dehydrogenase level increased by many 10 times; prothrombin time prolonged significantly. All the children were given antiviral therapy with intravenously injected acyclovir or ganciclovir, as well as hepatocyte growth factor to promote hepatocyte growth and hormone to alleviate inflammatory response. Two children were given plasma exchange in addition, among whom one was given the combination of continuous venovenous hemodiafiltration. Two children with HLH were given chemotherapy according to the HLH-2004 regimen. Three children survived, and one child with HLH died of multiple organ failure. It is concluded that EBV infection can cause acute liver failure and that early use of multimodality therapy including blood purification may be beneficial for prognosis in these children.
Child, Preschool ; Epstein-Barr Virus Infections ; Female ; Herpesvirus 4, Human ; Humans ; Infant ; Liver Failure, Acute ; Lymphohistiocytosis, Hemophagocytic ; Male ; Retrospective Studies

OBJECTIVEThe purpose of this study is to evaluate the possibility and surgical principle of titanium mesh used for the reconstruction of skull base bone defect.

METHODSThe clinical data of 11 patients with defect of skull base bone repaired with titanium mesh were retrospectively analysed.

RESULTSAmong 11 patients, there were 6 patients with skull base tumor, 3 patients with fibrosis hyperplasia, 2 patients with encephalomeningocele. The surgical approach included craniofacial approach in 7 patients, transfrontal and extended transfrontal approach in 3 patients, trans-midface approach in 1 patient. The anterior and lateral skull base was repaired in 2 patients, anterior and middle skull base and sellar repaired in 6 patients, anterior skull base and orbital floor repaired in 3 patients. In early postoperative period, there were 3 patients with intracranial pneumatosis, but without symptom, and 1 patient with transient cerebrospinal leakage. Following-up for average 14.4 months, there was no titanium mesh displacement and intracranial infection in all patients.

CONCLUSIONSThe titanium mesh used for the repair of skull base bone defect was both possible and safe.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Postoperative Period ; Reconstructive Surgical Procedures ; instrumentation ; methods ; Retrospective Studies ; Skull Base ; pathology ; surgery ; Surgical Mesh ; Titanium ; Young Adult

Objective To investigate the effect on the parameters of respiratory and circulatory systems in respiratory failure patients receiving mechanical ventilation by different suctioning. Methods 40 respiratory failure patients receiving mechanical ventilation were randomly divided into control group and experimental group (20 patients per group). Individuals in experimental group were treated with closed endotracheal suctioning and the others were treated with open endotracheal suctioning. The parameters of respiratory and circulatory systems were observed before and after suctioning. All the subjects used own control. Results The parameters of respiratory and circulatory systems did not change significantly in experimental group. The heart rate,SpO2,Ppeak,Pplat have changed significantly and the SBP,RR and PaO2 also reached the statistical significance before and after suctioning in control group. However,Vt and PaCO2 did not change significantly(P ＞ 0.05).Conclusions Respiratory failure patients treated with closed endotracheal suctioning was safer and more effective than traditional open-suctioning.

OBJECTIVETo evaluate fractional exhaled nitric oxide (FENO) level in patients with subacute cough and its value in predicting the patients' response to inhaled corticosteroids (ICS) treatment.

METHODSA total of 100 patients with persistent cough lasting more than 3 weeks were enrolled, including 52 patients with subacute cough and 48 with chronic cough. FENO, spirometry, and responses to ICS therapy of the patients were evaluated.

RESULTSThe recruited patients had a median (inter-quartile ranges) FENO level of 19 ppb (12-30 ppb). Patients with chronic cough had a significantly higher median FENO level than those with subacute cough (20.5 vs 16 ppb; Z=-2.245, P=0.025). A FENO level ≥25 ppb was recorded in 15 (28.8%) patients with subacute cough, as compared with 20 (41.6%) in patients with chronic cough (χ(2)=1.801, P=0.179). With a FENO ≥25 ppb as the critical value to justify ICS treatment, 15 patients with subacute cough received ICS and 14 (93.3%) of them showed obvious relief of cough after 2 weeks of therapy, a response rate similar to that of 85.0% (17/20) in patients with chronic cough receiving the treatment (χ(2)=0.588, P=0.443). In patients with subacute cough, those with cough variant asthma (CVA) or eosinophilic bronchitis (EB) had a significantly higher median FENO level than those with postinfectious cough [(16 (11-31) ppb vs 11 (8-19) ppb, P<0.01]. In the etiological analysis, CVA or EB was identified in 23 (44.2%) of the patients with subacute cough, as compared 21 (43.8%) in patients with chronic cough (χ(2)=0.002, P=0.961).

CONCLUSIONFENO may be an important indicator for etiological diagnosis of subacute cough and for predicting the response to ICS treatment.

Adrenal Cortex Hormones ; therapeutic use ; Breath Tests ; Chronic Disease ; Cough ; diagnosis ; drug therapy ; Exhalation ; Female ; Humans ; Male ; Nitric Oxide ; analysis

OBJECTIVETo investigate the association between rs9722 polymorphisms in the S100B gene and hand, foot and mouth disease (HFMD) caused by enterovirus 71.

METHODSA total of 124 HFMD children with enterovirus 71 infection were enrolled as subjects, and 56 healthy children were enrolled as control group. The rs9722 polymorphisms in the S100B gene were detected for both groups, and the serum level of S100B protein was measured for 74 HFMD children.

RESULTSThe rs9722 locus of the S100B gene had three genotypes, CC, CT, and TT, and the genotype frequencies were in accordance with Hardy-Weinberg equilibrium. Compared with the control group, the HFMD group had significant increases in the frequencies of TT genotype and T allele (P<0.01). Children with severe HFMD caused by enterovirus 71 infection had significantly higher frequencies of TT genotype and T allele than those with moderate or mild HFMD (P<0.05). Compared with the cured patients, the patients with poor prognosis had significant increases in the frequencies of TT genotype and T allele in the rs9722 locus of the S100B gene (P<0.05). Among the 74 children with HFMD, the children with TT genotype had the highest serum level of S100B protein, and those with CC genotype had the lowest level (P<0.01).

CONCLUSIONST allele in the rs9722 locus of the S100B gene might be a risk factor for severe HFMD caused by enterovirus 71 infection.

Child, Preschool ; Enterovirus A, Human ; Enterovirus Infections ; complications ; Female ; Genotype ; Hand, Foot and Mouth Disease ; etiology ; genetics ; Humans ; Infant ; Male ; Polymorphism, Genetic ; S100 Calcium Binding Protein beta Subunit ; genetics