Drug Contamination ; Drugs, Chinese Herbal ; analysis ; Fourier Analysis ; Plants, Medicinal ; chemistry ; Spectrum Analysis, Raman ; instrumentation ; methods

OBJECTIVETo study the relationship between HBeAg expression and HBV-DNA in serum and peripheral blood mononuclear cells (PBMCs).

METHODS208 patients with chronic hepatitis B were included in this present study. HBV-DNA in the PBMCs were performed by polymerase chain reaction (PCR), with the serum HBV-DNA level being determined by the way of fluoresces quantities PCR (FQ-PCR). Meanwhile, HBV-GM was also detected via enzyme-linked immunosorbent assay (ELISA).

RESULTSThere were 106 patients for positivity in the HBV-DNA level of PBMCs with 102 for negativity, in which the HBV-DNA high levels (HBV DNA load > or = 1.0E5) in serum were 91.5%, 45.1% (chi2=52.12, P>0.01) respectively, with 76.4% and 50.9% (chi2=21.55, P>0.01) for the positive percentage of HBeAg expression.

CONCLUSIONA significantly positive correlation was found between HBV-DNA in PBMCs and serum HBV-DNA along with the positive percentage of HBeAg, indicating that obvious PBMCs' increase infected by HBV in patients with positivity of HBeAg and high level of serum HBV-DNA.

Adolescent ; Adult ; Aged ; Antibodies, Viral ; blood ; DNA, Viral ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Hepatitis B ; genetics ; immunology ; Hepatitis B e Antigens ; immunology ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Leukocytes, Mononuclear ; virology ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Young Adult

OBJECTIVETo analyze the clinical characteristics of plastic bronchitis associated with 2009 influenza A virus (H1N1) infection.

METHODA retrospective investigation of the clinical manifestation, bronchoscopy, and the histology of the cast, clinical course and outcome of 8 children with plastic bronchitis associated with influenza A virus (H1N1) infection during winter of 2009 and 2010 was performed.

RESULTAll 8 cases were boys, the range of age was 3 to 6 years. Five cases occurred in 2009 winter, accounting for 3.3% (5/150) of hospitalized children with influenza A (H1N1) infection; 3 cases occurred in 2010 winter, accounting for 15.8% (3/19) of hospitalized children with influenza A (H1N1) infection. Two patients had an underlying chronic disease, 1 had asthma, and the other had allergic rhinitis and atopic dermatitis. All the 8 cases had fever, cough and sputum; 2 had wheezing; 5 had respiratory distress. All 8 cases were diagnosed as influenza A virus (H1N1) infection complicated with pneumonia, of whom 5 patients had atelectasis, 2 had pneumothorax, 1 had pneumomediastinum, 1 had parapneumonic effusion, 2 patients were suspected of foreign body aspiration. Seven cases were admitted to an ICU, 5 patients developed respiratory failure, and 3 patients required mechanical ventilation. Flexible bronchoscopy and bronchial lavage was performed in all cases and showed bronchial cast. Histological examination of the bronchial cast revealed a fibrinous material containing large quantity of eosinophils, neutrophils, and lymphocytes in 7 patients, fibrinous material and necrotic material without inflammatory cells in 1 patient. After the bronchial cast was removed, all patients were improved greatly, no patients died.

CONCLUSIONPlastic bronchitis is a life-threatening complication associated with 2009 influenza A (H1N1) virus infection in children. In children with rapid and progressive respiratory distress with lung atelectasis or consolidation on chest radiograph, plastic bronchitis should be considered. Bronchoscopic extraction of casts should be carried out early.

Antiviral Agents ; administration & dosage ; therapeutic use ; Bronchitis ; complications ; diagnosis ; therapy ; virology ; Bronchoscopy ; Child ; Child, Preschool ; Foreign Bodies ; complications ; Glucocorticoids ; administration & dosage ; therapeutic use ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; complications ; virology ; Intensive Care Units ; Male ; Pulmonary Atelectasis ; diagnosis ; therapy ; virology ; Rare Diseases ; Respiratory Insufficiency ; diagnosis ; therapy ; virology ; Retrospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

OBJECTIVETo explore the possible immuno-regulating mechanism of action of Chinese drugs in different combinations (assembled depending different therapeutic principles) through observing the effects of Feiliuping ointment (FLP) and its disassembled prescriptions on dendritic cells (DC) in blood, spleen and tumor in mice with transplanted Lewis lung cancer (LLC).

METHODSPercentages of DC in blood, spleen and tumor of mice with transplanted LLC treated by FLP and its disassembled prescriptions were estimated, and the S-100 protein expression in tumor tissue was detected by immunohistochemical method.

RESULTSThe percentage of DC (per thousand) in tumor bearing mice was 0.43 +/- 0.26 in peripheral blood, and 0.32 +/- 0.16 in spleen, significantly lower than those in normal mice 4.68 +/- 0.90 and 3.68 +/- 1.58, P<0.01); and S-100 protein expression in tumor was weakened. After FLP treatment, the percentages of DC (per thousand) in tumor bearing mice were increased to 2.55 +/- 0.29 in peripheral blood and 2.70 +/- 0.63 in spleen (P<0.01), with the S-100 protein expression in tumor tissue up-regulated significantly (P<0.01). Study on different assembled prescriptions of FLP showed that the qi supplementing components of FLP displayed the optimal actions.

CONCLUSIONFLP, a Chinese herbal prescription made depending on Chinese medicine therapeutic principle of strengthening body resistance and consolidating constitution, has an obvious anti-tumor effect, to improve the immunological anti-tumor function of organism by promoting the amount and expression of DC might be the possible intrinsic mechanism.

Animals ; Carcinoma, Lewis Lung ; immunology ; pathology ; Cell Line, Tumor ; Dendritic Cells ; drug effects ; immunology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Phytotherapy ; S100 Proteins ; metabolism

Objective To investigate the clinical characteristics of plastic bronchitis (PB) so as to improve the awareness of the disease.Methods Twenty-four children with PB were collected from Jul.2009 to Mar.2012 in Shenzhen Children's Hospital.The clinical manifestation,bronchoscopy,histology of the cast,clinical course and outcome were reviewed retrospectively.Results Of the 24 children with PB,18 cases were male,6 cases were female,and the range of age was 1 year and 2 months to 10 years and 3 months,with the median age of 3 years and 4 months.Three patients had an underlying chronic disease,1 case had asthma,1 case had hydronephrosis,and 1 case had ventricular septal defect repair before 1 year and 8 months.All the cases had fever,cough and sputum,while 10 cases had wheeze,and 5 cases had respiratory distress.All cases were diagnosed as pneumonia or severe pneumonia,of which 14 case had atelectasis,10 cases had parapneumonic effusion,5 cases suspected of foreign body inhalation,3 cases had pneumothorax,and 3 cases had mediastinal hernia.Fourteen cases were admitted to PICU,6 patients developed respiratory failure,and 9 patients required mechanical ventilation.Flexible bronchoscopy and bronchial lavage were performed in all cases and showed bronchial cast.Histological examination of the bronchial cast revealed that fibrinous material containing large quantity of eosinophils,neutrophils,and lymphocytes in 23 patients,and no inflammatory cells in 1 patient.After a bronchial cast was removed,all patients were improved greatly,and no patient dead.Conclusions Plastic bronchitis is a rare pediatric critical disease,which has high mortality.In children with rapid and progressive respiratory distress with lung atelectasis,pleural effusion or consolidation on chest radiograph,PB should be considered.Bronchial endoscopy is the most effective method for treatment of PB.

Objective: To investigate the effect and related mechanism of homocysteine (Hcy) on calcium overload in neonatal rat atrial cells (NRICs). Methods: NRICs were assigned to 9 groups after culture for 3 days: (1) control group; (2) Hcy group (0, 50, 100, 200, 500 μmol/L for 48 hours); (3) antioxidant group (NAC, 10 μmol/L for 24 hours); (4) Hcy+NAC group (500 μmol/L Hcy for 48 hours, then treated with 10 μmol/L NAC for 24 hours); (5) calcium/calmodulin dependent protein kinase Ⅱδ (CaMKⅡδ) inhibitor group (KN-93, 3 μmol/L KN-93 for 5 hours); (6) specific sodium current inhibitor group (ELE, 1 μmol/L ELE for 5 hours); (7) Hcy+KN-93 group (500 μmol/L Hcy for 48 hours, then treated with 3 μmol/L KN-93 for 5 hours); (8) Hcy+ELE group (500 μmol/L Hcy for 48 hours, then treated with 1 μmol/L ELE for 5 hours; (9) Hcy+KN-93+ELE group (500 μmol/L Hcy for 48 hours, then treated with 3 μmol/L KN-93 and 1 μmol/L ELE for 5 hours). Moreover, NRICs were also treated with CaMKⅡδ-siRNA lentivirus, and Nav1.5-siRNA lentivirus, negative lentivirus carrier containing green fluorescent protein (GFP) for 24 hours. The MOI values of the three groups were 10. Infection efficiency of lentivirus was determined by observing the percentage of GFP fluorescence under inverted fluorescence microscope after transfection for 24 hours, and cultured regularly with simultaneous Puro screening, then cells were grouped as Hcy+CaMKⅡδ-siRNA group, Hcy+Nav1.5-siRNA group and Hcy+negative group. The concentration of Ca²⁺ in NRICs ([Ca²⁺]i) of various groups was detected through Fluo-4/AM fluorescence probe, then 2', 7'- two chlorofluorescein diacetate (DCFH-DA) was used as a probe to detect reactive oxygen species (ROS) in NRICs by flow cytometry. The malondialdehyde (MDA) was detected by the activity of superoxide dismutase (SOD) and xanthine oxidase was detected by thiobarbituric acid colorimetry. The protein and mRNA expression level of CaMKⅡδ and Nav1.5 in NRICs were detected by Western blot and quantitative real-time PCR. Results: (1) ROS, MDA and SOD were similar between NAC group and control group, ROS and MDA were significantly increased, while SOD was significantly reduced in Hcy group in a concentration-dependent manner. (2) [Ca²⁺]i: The level of [Ca²⁺]i was (155.57+7.25), (187.43+13.07), (248.98+27.22) and (307.36+15.09) nmol/L in 50, 100, 200 and 500 μmol/L Hcy groups, which was significantly higher than that in the control group ((123.18+7.24) nmol/L, P<0.01). In addition, the level of [Ca²⁺]i in Hcy+NAC group ((222.87+23.71)nmol/L) was significantly lower than that in Hcy 500 μmol/L group ((305.15+39.45) nmol/L, P<0.05), while [Ca²⁺]i level was similar between NAC group and the control group. (3) The protein expression of CaMKⅡδ and Nav1.5 was significantly upregulated in Hcy groups than in the control group. The protein expression level of CaMKⅡδ-Thr287 was significantly lower in NAC group than in Hcy 500 μmol/L group (P<0.01), however, there was no significant difference on the protein expression levels of CaMKⅡδ-Thr287 and Nav1.5 between NAC group and control group (all P>0.05). (4) The protein expression levels of CaMKⅡδ-Thr287 and the concentration of [Ca²⁺]i were significantly lower in Hcy+KN-93 group and Hcy+KN-93+ELE group than in Hcy 500 μmol/L group (P<0.05). [Ca²⁺]i concentration was significantly lower in Hcy+KN-93 group, Hcy+ELE group and KN-93+ELE+Hcy group than in Hcy 500 μmol/L group (P<0.05). (5) The mRNA and protein expression levels of CaMKⅡδ and Nav1.5 in each group infected with lentivirus: the GFP expression was ideal post lentivirus transfection for 24 hours (up to 90%), which was significantly lower in the CaMKⅡδ-siRNA group and Nav1.5-siRNA group than in the negative infection group (all P<0.05), which was similar between negative infection group and control group (P>0.05). Moreover, the mRNA and protein expression levels of CaMKⅡδ and CaMKⅡδ-Thr287 was significantly lower in Hcy+Nav1.5-siRNA group than in Hcy+negative infection group (P<0.05). The protein and mRNA levels of Nav1.5 were similar between Hcy+CaMKⅡδ-siRNA group and Hcy+negative infection group (P>0.05). Conclusions Hcy can induce calcium overload in NRICs by increasing oxidative stress, upregulating the sodium channel protein, and activating the late sodium current and phosphorylating CaMKⅡδ.

OBJECTIVETo investigate the expression of special AT-rich sequence binding protein 1 (SATB1) mRNA in hepatocellular carcinoma (HCC) and explore its correlation to the clinicopathological features, surgical outcomes and metastasis of HCC.

METHODSThe total RNA was extracted from 102 HCC tissues and the adjacent tissues, and the expression of SATB1 mRNA was detected using quantitative real-time PCR. The correlations of SATB1 mRNA expression to the clinicopathological features, postoperative recurrence and metastasis of the tumor were analyzed.

RESULTSThe expression of SATB1 mRNA in HCC tissues was 3.27 folds higher than that in the adjacent tissues (P<0.001). The expression of SATB1 mRNA in HCC was associated with liver cirrhosis, AFP level, tumor size, tumor thrombi, histological differentiation, TNM classification, postoperative recurrence and metastasis (P<0.05), but not to the patients' gender, age, HbsAg positivity, HCV-Ab positivity, tumor number, or the presence of tumor encapsulation (P>0.05). In patients with significant high expression, high expression, and low expression of SATB1 mRNA, the postoperative recurrence rates were 82.68%, 0, and 0, with the 3-year survival rate of 0, 52.63%, and 100%, respectively.

CONCLUSIONSATB1 mRNA expression is associated with the postoperative recurrence and metastasis of HCC, and can be used as an indicator for predicting the recurrence and metastasis of HCC.

Aged ; Carcinoma, Hepatocellular ; genetics ; metabolism ; Female ; Humans ; Liver Neoplasms ; genetics ; metabolism ; Male ; Matrix Attachment Region Binding Proteins ; genetics ; metabolism ; Middle Aged ; Neoplasm Metastasis ; diagnosis ; Neoplasm Recurrence, Local ; diagnosis ; RNA, Messenger ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; methods

OBJECTIVETo explore suitable scaffold material for big segmental long bone defect by studying the properties of the prepared deproteinized bone.

METHODSCancellated bone were made as 30 mm x mm x 3 mm bone blocks from inferior extremity of pig femur along bone trabecula. The deproteinized bone was prepared with an improved method. Their morphological features, components, cell compatibility, mechanical and immunological properties were investigated respectively.

RESULTSDeproteinized bone maintained natural reticular pore system. The main organic material is collagen I and inorganic composition is hydroxyapatite. It has good mechanical properties, cell adhesion rate and histocompatibility.

CONCLUSIONThis deproteinized bone can be applicable as scaffold for reparation of big segmental defect in long bone.

Animals ; Bone Regeneration ; physiology ; Bone Transplantation ; methods ; Collagen ; Hydroxyapatites ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo investigate the implications of erythroblasts periodic acid-Schiff (PAS) stain for myelodysplastic syndromes (MDS) dyserythropoiesis, diagnosis and differential diagnosis.

METHODSPAS stain of bone marrow (BM) erythroblasts in 406 MDS patients, 207 non-severe aplastic anemia (NSAA), 144 immune thrombocytopenic purpura (ITP), 67 megaloblastic anemia (MegA), 76 iron deficiency anemia (IDA), 50 paroxysmal nocturnal hemoglobinuria (PNH), and 50 acute erythroid leukemia (AEL) as well as some related laboratory parameters in MDS patients were analyzed retrospectively.

RESULTSPAS-positive detection rate was significantly higher in MDS (53.0%) than in NSAA (14.5%), ITP (27.1%) and PNH (16.0%), but was significantly lower in MDS than in AEL (84.0%) (all P = 0.000). There was no significant difference in PAS-positive detection between MDS and MegA (46.3%), or MDS and IDA (40.8%) (P = 0.310, 0.052, respectively). Erythroblasts PAS-positive rate (Median, M = 1%) and PAS-positive scores (M' = 2) was significantly lower in MDS than in AEL (M = 8%; M' = 17), and significantly higher than in NSAA (M = 0%; M' = 0), ITP (M = 0%; M' = 0), PNH (M = 0%; M' = 0), MegA (M = 0%; M' = 0), and IDA (M = 0%; M' = 0) (all P < 0.05). The cut-off value of PAS-positive rate and score for distinguishing MDS from the other groups except AEL were 0.5% and 0.5, with a sensitivity and specificity of 60.8% and 74.4%, respectively. For MDS patients, the percentage of BM erythroid cells was significantly higher in PAS-positive group than in PAS-negative group (P < 0.05), and so were megakaryocyte count, lymphocyte-like micromegakaryocytes count and percentage of micromegakaryocyte (P = 0.002, 0.000, 0.000, respectively). HGB, MCV, MCH and MCHC were significantly lower in PAS-positive group (all P < 0.05), and so was the neutrophil alkaling phosphatase (NALP) (P = 0.000). PAS-positive detection rate, positive rate and score were higher in MDS patients with abnormal karyotype than with normal karyotype, and were also higher in IPSS high/intermediate-risk 2 group than in low/intermidiate-risk 1 group.

CONCLUSIONThe positive reaction of erythroblasts PAS stain is an indicator of dyserythropoiesis. It is helpful to the diagnosis of MDS patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Diagnosis, Differential ; Erythroblasts ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; diagnosis ; Periodic Acid-Schiff Reaction ; Retrospective Studies ; Sensitivity and Specificity ; Staining and Labeling ; Young Adult

OBJECTIVETo study biomechanical changes of newly formed bones 24 weeks after repairing large defects of long bones of goats using heterogeneous deproteinated bone (DPB) prepared by modified methods as an engineering scaffold.

METHODSAccording to a fully randomized design, 18 goats were evenly divided into three groups: normal bone control group (Group A), autologous bone group (Group B) and experimental group (Group C). Each goat in Groups B and C were subjected to the periosteum and bone defect at middle-lower part of the right tibia (20% of the whole tibia in length), followed by autologous bone or DPB plus autologous MSCs + rhBMP2 implantation, respectively and semi-ring slot fixation; while goats in Group A did not perform osteotomy. At 24 weeks after surgery, biomechanical tests were carried out on the tibias.

RESULTSAt 24 weeks after surgery, the results of anti-compression test on tibias in three groups were recorded by a functional recorder presented as linear pressure-deformation curve. The shapes of the curves and their change tendency were similar among three groups. The ultimate pressure values were 10.74 MPa+/-1.23 MPa, 10.11 MPa+/-1.35 MPa and 10.22 MPa+/-1.32 MPa and fracture compression rates were 26.82%+/-0.87%, 27.17%+/-0.75% and 28.22%+/-1.12% in Groups A, B and C, respectively. Comparisons of anti-compression ultimate pressures and fracture compression rates among three groups demonstrated no significant difference (P(AB) equal to 0.415, P(BC) equal to 0.494). Three-point anti-bend test on tibias was recorded as load-deformation curves, and the shapes of the curves and their change tendency were similar among three groups. The ultimate pressure values of the anti-bend test were 481.52 N+/-12.45 N, 478.34 N+/-14.68 N and 475.62 N+/-13.41 N and the fracture bend rates were 2.62 mm+/-0.12 mm, 2.61 mm+/-0.15 mm and 2.81 mm+/-0.13 mm in Groups A, B and C, respectively. There was no significant difference between groups (P(AB) equal to 0.7, P(BC) equal to 0.448). The ultimate anti-torsion torque values were 6.55 Nm+/-0.25 Nm, 6.34 Nm+/-0.18 Nm and 6.42 Nm+/-0.21 Nm and fracture torsion rates were 29.51 degree+/-1.64degree, 28.88 degree+/-1.46 degree and 28.81 degree+/-1.33 degree in Groups A, B and C, respectively. There was no significant difference between groups (P(AB) equal to 0.123, P(BC) equal to 0.346).

CONCLUSIONSThe biomechanical characteristics of newly formed bones from heterogeneous DPB for repairing large segmental long bone defect are comparable to those of normal bones and autologous bones. DPB has the potential for clinical usage as bone graft material.

Animals ; Biomechanical Phenomena ; Female ; Goats ; Male ; Proteins ; Tibia ; physiology ; surgery ; Tissue Engineering ; Torsion, Mechanical