Obesity is increasingly prevalent globally, searching for therapeutic agents acting on adipose tissue is of great importance. Equisetin (EQST), a meroterpenoid isolated from a marine sponge-derived fungus, has been reported to display antibacterial and antiviral activities. Here, we revealed that EQST displayed anti-obesity effects acting on adipose tissue through inhibiting adipogenesis in vitro and attenuating HFD-induced obesity in mice, doing so without affecting food intake, blood pressure or heart rate. We demonstrated that EQST inhibited the enzyme activity of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), a therapeutic target of obesity in adipose tissue. Anti-obesity properties of EQST were all offset by applying excessive 11β-HSD1's substrates and 11β-HSD1 inhibition through knockdown in vitro or 11β-HSD1 knockout in vivo. In the 11β-HSD1 bypass model constructed by adding excess 11β-HSD1 products, EQST's anti-obesity effects disappeared. Furthermore, EQST directly bond to 11β-HSD1 protein and presented remarkable better intensity on 11β-HSD1 inhibition and better efficacy on anti-obesity than known 11β-HSD1 inhibitor. Therefore, EQST can be developed into anti-obesity candidate compound, and this study may provide more clues for developing higher effective 11β-HSD1 inhibitors.

Objective: To establish a better near infrared quantitative model for quality control of Glycyrrhizae Radix et Rhizoma of components (moisture,total ash,liquiritin and glycyrrhizic acid) in liquorice,in order to realize rapid detection.Method: The contents of moisture,total ash,liquiritin and glycyrrhizic acid were determined in 97 samples based on the methods set forth in Chinese Pharmacopoeia.Meanwhile,the near infrared spectrum was scanned using near infrared spectroscope.R software was used to screen out the spectral pretreatment and build the quantitative models.Result: The optimum spectral pretreatment method for establishing the near infrared quantitative model of moisture and liquiritin was the first order derivative.For moisture,the correlation coefficients of test and validation were 0.930 0 and 0.929 9,and the root mean square errors were 0.243 2 and 0.203 8,respectively.For liquiritin,the correlation coefficients of test and validation were 0.930 3 and 0.907 6,and the root mean square errors were 0.093 9 and 0.128 9,respectively.The optimum spectral pretreatment method for establishing the near infrared quantitative model of total ash was MSC.The correlation coefficients of test and validation were 0.926 5 and 0.917 7,and the root mean square errors were 0.109 6 and 0.103 7,respectively.The optimum spectral pretreatment method for establishing the near infrared quantitative model of glycyrrhizic acid was SNV.The correlation coefficients of test and validation were 0.918 1 and 0.915 7,and the root mean square errors were 0.274 8 and 0.236 0,respectively.Conclusion: In this study,a better near infrared quantitative models for quality control of components of Glycyrrhizae Radix et Rhizoma were established,with a high accuracy,which laid a foundation for rapid detection of the components in Glycyrrhizae Radix et Rhizoma.

OBJECTIVETo investigate the effects of Btk inhibitor (PCI-32765) and BCR-ABL tyrosine kinase inhibitor (Dasatinib) on proliferation and apoptosis of acute lymphoblastic leukemia (ALL) cell lines (Sup-B15, RS4;11) and the possible mechanism.

METHODSRS4;11 and Sup-B15 cells were treated with PCI-32765 and Dasatinib, the cell proliferation and apoptosis were detected by CCK-8, the Btk and other apoptotic proteins were detected by Western blot.

RESULTSPCI-32765 could inhibit the proliferation of RS4;11 and Sup-B15 cells in a dose-dependent manner, Sup-B15 cells were more sensitive to PCI-32765 than RS4;11 cells, their ICwere 3 µmol/L and 8 µmol/L respectively, the difference between them was statistically significant (P<0.05). Dasatinib also could inhibit the proliferation of RS4;11 cells and Sup-B15 cells in a dose-dependent manner. The ICwas 5 µmol/L and 5 nmol/L, respectively, the difference between them was statistically very significant (P<0.01), and the inhibitory effect was enhanced by the combination of Damatinib with the PCI-32765(P<0.05). The cell survival rate decreased gradually in PCI-32765 or Dasatinib alone group and the combination group at the different time-point (8, 12, 24, 36, 48 and 72 h), the 2 drugs showed a synergistic effect on cells in a time-dependent manner. After being treated with PCI-32765 and Dasatinib, the RS4;11 and Sup-B15 cells showed that cell shrinkage, increase of cytoplasmic density, nuclear pyknosis, deviation and karyorrhexis, and increase of the apoptotic cells in the combination group, while the promotive effect of low dosage dasatinib on apoptosis of RS4;11 cells was not strong. PCI-32765 and Dasatinib could decrease the expression and activity of BCR-ABL, Btk, Lyn, Src in Sup-B15 and RS4;11 cells.

CONCLUSIONPCI-32765 or Dasatinib can inhibit the proliferation and induce the apoptosis of Sup-B15 and RS4;11 cells, PCI-32765 and Dasatinib displayed the synergistic effects. The possible mechanism may be related with the blocking of B cell receptor(BCR) signal pathway, thereby inhibiting the cell proliferation and promoting the cell apoptosis.

OBJECTIVETo investigate the inductive therapeutic effects of imatinib combined with VP low dose regiment on adult patients with Ph-positive acute lymphoblastic leukemia (Ph(+) ALL).

METHODSFourteen newly diagnosed adult patients with Ph(+) ALL were treated with VP regimen, and imatinib (400 mg/d) was added at the 8(th) day. VP regimen would be stopped when neutropenia lasted for 1 week or complicated with infection, fever, etc. Therapeutic effects were assessed by bone marrow morphology and quantitative analysis of BCR/ABL:ABL at the 28(th) - 33(rd) day. Patients could be treated with imatinib combined with chemotherapy for consolidation and maintenance therapy or were treated with allogeneic hematopoietic stem cell transplantation after complete remission.

RESULTSFourteen cases obtained CR1 after first course of treatment, the median decline of BCR/ABA:ABL was 55.89 (10.25 -180.97) %; during the induction chemotherapy, pulmonary infection occurred in 3 patients, diarrhea in 1 patients, facial edema in 3 patients, however, all these patients were cured after symptomatic treatment, only 1 patient died of relapse after transplantation.

CONCLUSIONIn the period of tyrosine kinase inhibitor (TKI), inductive chemotherapy combined with imatinib and low dose VP can obtaine satisfactory CR rate and decrease the toxicity of the traditional drugs. It is suggested that TKI combined with VP regimen chemotherapy can achieve CR1 and make possible for allo-HSCT, from which patients can achieve the long-term survival.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; Bone Marrow ; Cisplatin ; Fusion Proteins, bcr-abl ; Hematopoietic Stem Cell Transplantation ; Humans ; Imatinib Mesylate ; Induction Chemotherapy ; Neutropenia ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Protein Kinase Inhibitors ; Recurrence ; Remission Induction ; Transplantation, Homologous ; Vindesine

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.

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

The aim of this study is to develop the recombinant adenovirus vaccine (rAdV) candidates containing neuraminidase (NA) gene of H5N1 influenza virus and test in BALB/c mice the effect of cell-mediated immunity. In this study, two kind of NA gene (WtNA gene, the wild type; Mod. NA gene, the codon-modified type) derived from H5N1 influenza virus (A/Anhui/1/2005) were cloned and inserted respectively into plasmid of adenovirus vector, then the rAdV vaccines candidates (rAdV-WtNA and rAdV-Mod. NA) were developed and purified, followed by immunization intramuscularly (10(9) TCID50 per dose, double injection at 0 and 4th week) in BALB/c mice, the effect of cell-mediated immunity were analysed at 5th week. Results indicated that: (i) NA protein expression was detected in two rAdV vaccines candidates by Western blotting; (ii) the rAdV-Mod. NA vaccine could elicit more robust NA specific cell-mediated immunity in mice than that of rAdV-WtNA vaccine (P = 0. 016) by IFN-gamma ELIspot assay. These findings suggested rAdV-Mod. NA vaccine was a potential vaccine candidate against H5N1 influenza and worthy of further investigation.
Animals ; Blotting, Western ; Female ; Humans ; Immunity, Cellular ; genetics ; immunology ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Influenza Vaccines ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Neuraminidase ; genetics ; immunology ; Orthomyxoviridae Infections ; immunology ; virology ; Polymerase Chain Reaction ; Random Allocation ; Viral Proteins ; genetics ; immunology

Objective To investigate immunity of a recombinant adenovirus vaccine(rAdV)containing codon-modified neuraminidase(Mod.NA)gene of H5N1 influenza virus in BALB/c mice and to screen for appropriate dose.Methods BALB/c mice were immunized with the rAdV-Mod.NA vaccine intramuscularly twice (double injection at 0 and 4~(th) week)in three groups,low dosage(10~5 TCI_(50) per dose),medium dosage(10~7 TCID_(50) per dose)and high dosage(10~9 TCID_(50) per dose).The effect of humoral and cell-mediated immunity were analysod at 5~th week.Results ①The rAdV-Mod.NA vaccine eould elicit both humoral and cell-mediated robust NA specific immunity in mice by neuraminidase inhibitor assay and IFN-γ ELISpot assay;②10~7 TCID_(50) per dose Was the appropriate dose;③ Peptide NA_(109-124):CRTFFLTQGALLNDKH and peptide NA_(182-199):AVAVLKYNGIITDTIKSW were the dominant epitopes for neuraminidase-immunized BALB/c mice,which was screened out from the whole length of neuraminidase of an H5N1 virus,A/Anhui/1/2005.Conclusion The recombinant adenovirus NA could induce specific humoral and cellular immune responses in BALB/c after immunization,which suggest rAdV-Mod.NA vaccine was a potential vaccine candidate against H5N1 influenza and worthy of further investigation.

In order to improve the expression of human avian influenza virus hemagglutinin (HA) and meet the pandemic influenza vaccine needs, we optimized and synthesized the whole length of HA gene of H5N1 (A/Anhui/1/2005) influenza virus in accordance with the human's codon preference, and inserted it to the eukaryotic expression vector pDC315 to construct a eukaryotic expression vector pDC315-Mod. HA. This plasmid and the eukaryotic expression vector pDC315-Wt. HA containing wild HA gene were transfected into 293T cells respectively to compare the expression of HA protein. The results showed that according to the comparison and identification by indirect immunofluorescence assay and Western blot test, the expression of HA protein in 293T cells was significantly improved after codon optimization. This laid a foundation for pandemic influenza vaccine research.
Blotting, Western ; Cell Line ; Codon ; Fluorescent Antibody Technique, Indirect ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; Plasmids

Objective To investigate the influence of avian influenza virus (AIV) NS1 protein on the expression of interferon-inducible protein 10 (IP-10). Methods NS1 gene from virus A/Anhui/1/2005 (H5N1),NS1 gene inserted with 80-84 amino acids from virus A/Anhui/1/2005(H5N1)and NS1 gene from virus A/Puerto Rico/8/1934 (H1N1) were cloned into the eukaryotic expression vector pEGFP-N1, and transected into BEAS-2Bcells, IP-10 expression level in transected cells was detected by flow cytometry. Results Compared with the control group pEGFP-N1, Expression of these three different NS1 genes can down-regulate the expression of IP-10in BEAS-2B cells, but there is no significant difference as to the lower level among them. Conclusion NS1protein of A/Anhui/1/2005(H5N1) can down-regulate the expression level of IP-10, but this may not clarify its relationship with the virulence of AIV.