BACKGROUND:Hyperbaric oxygen(HBO)treatment promotes the proliferation and differentiation of endogenous neural stem cells in neonatal rats following hypoxic/ischemic brain damage(HIBD).The Wnt signaling pathway is associated with neurogenesis.However,there are few data recording the role of HBO in the differentiation of neural stem cells in vitro.OBJECTIVE:To observe the effect of HBO on differentiation and Wnt3 expression of bone marrow mesenchymal stem cells(BMSCs).METHODS:BMSCs were isoiated and cultured.The rat BMSCs of passages 3-5 were cultured in DMEM/F12(1:1)medium with basic fibroblast growth factor,epidermal growth factor and B27 for 24 hours.The induced BMSCs were randomly divided into two groups:control group(no treatment)and HBO group(HBO,0.10 MPa,60 minutes stabilizing pressure with at least 90% oxygen).The neuron specific encloase(NSE),glial fibrillary acidic protein(GFAP)and 04 marked oligodendrocyte immunocytochemistry were detected by immunofluorescent staining,and Wnt3 protein expression was detected by Western-blot.RESULTS AND CONCLUSION:BMSCs cultured in classic medium of neural stem cells could significantly induce the expression of nestin.The expression of NSE and 04 of HBO group was greater than control group(P＜0.01),but GFAP expression displayed no significant difference between the groups(P＞0.05).Western blot showed HBO could enhance the Writ3 expression (P＜0.05).Results show that HBO can induce BMSCs to differentiate into neural cells and oligodendrocyte,which is correlated with the activation of the Wnt3 protein.

Objective To investigate the clinical value of homocysteine(Hcy) ,coagulation function ,platelet parameters and he-morheology detection in patients with cerebral infarction .Methods 114 patients with cerebral infarction were served as cerebral in-farction group ,while 112 healthy people as control group .Their platelet parameters ,coagulation function ,Hcy and hemorheology were detected .Results Hcy ,Fibrinogen(Fib) and D-dimer(D-D) of patients in cerebral infarction group were higher than those in control group(P<0 .05) ,while the differences of prothrombin time(PT) ,activated partial thromboplastin time(APTT) and throm-bin time(TT) between the two groups showed no statistically significant (P> 0 .05) .In cerebral infarction group ,platelet count (PLT) of patients was lower ,and platelet distribution width(PDW) and mean platelet volume(MPV) were higher than those in the control group(P<0 .05) while plateletcrit(PCT) showed no statistically significant difference between the two groups (P<0 .05) . Whole blood viscosity ,plasma viscosity and hematocrit of patients in the cerebral infarction group were higher than those in the con-trol group(P< 0 .05) .Conclusion Hcy ,coagulation function ,platelet parameters and hemorheology detection have important significance for prediction and early diagnosis and treatment of cerebral infarction .

This study was aimed to establish the quality standard of Pyrethri Tatsienenis Flos. The medical material was identified by the microscopy and the thin layer chromatography ( TLC ) methods . The moisture , total ash , acid-insoluble ash and alcohol-soluble extract were determined according to procedures recorded in the Chi-nese Pharmacopoeia (2010 edition). The content of luteolin was determined by the HPLC method. The results showed a strong characteristic microscopic of Pyrethri Tatsienenis Flos , and its TLC identification had a good resolution with clear spots . The mass fractions of luteolin was 0 . 036%~0 . 104% ( average of 0 . 078%) , moisture was 9 . 32%~15 . 82% ( average of 13 . 11%) , total ash was 6 . 65%~8 . 29% ( average of 7 . 45%) , acid-insoluble ash was 0 . 23%~0 . 59% ( average of 0 . 42%) , and the extraction was 21 . 42%~30 . 15% ( average of 24 . 86%) . It was concluded that this established standard was simple to operate with good stability and reproducibility , which can be used for quality evaluation of Pyrethri Tatsienenis Flos .

Objective To amplify the encoding full-length sequence of Tibet mini-pigs myostatin ( MSTN) gene and analysis the sequence by bioinformatics software.Method The RNA of liver tissues from Tibet mini-pig was extracted, and reversely transcribed into cDNA.The gene coding region sequence of myostatin gene was amplified through RT-PCR, and then the purified product of PCR was ligated with a pMD19-T and transferred into the bacterium DH5αfor replication.The positive clones were screened and sequenced. The sequence characters were analyzed by using bioinformatics method and phylogeny evolution tree was constructed with other twelve species.Results The coding redion of MSTN gene was 1128bp, and coded 375 amino acids.The amino acid homology analysis showed that the homology rate of amino acid sequence was 99%.Conclusions Molecular phylogeny evolution indicated that it had a close relation with human, dog, banna minipig, sheep, goat, cattle, horse, chimpanzee, rat, mouse except chick and zebrafish, and the most closely related with banna minipig.

Objective To distinguish residual tumor from inflammation after radiofrequency ablation (RA) for hepatic VX2 carcinoma in rabbits according to the comparative study between CT and pathological findings.Methods CT and pathologic examination were performed in different stages of RFA for rabbits hepatic VX2 models,and their different performances were observed.Results Marginal enhancement band was showed with enhanced CT of both residual tumor and inflammation.Moreover,liver tissues peripheral to enhancement band were in gradual weaken pattern.The enhancement band of inflammation was most obvious on the 2~(nd) day after RFA,but weakened gradually and disappeared two weeks later.Conclusion The residual tumor and inflammation could not be distinguished through enhanced CT scanning within 1 week after RFA.Low intensity lesions with peripheral enhancement 2 weeks after RFA should be recognized as residual tumor.

BACKGROUND:The enamel matrix derivative has been used in the clinical treatment of severe periodontitis; however, the mechanism(s) by which enamel matrix derivative promotes periodontal regeneration is stil obscure. OBJECTIVE:To explore the effects of enamel matrix derivatives on the differentiation and proliferation of periodontal ligament stem cels. METHODS:Periodontal ligament stem cels were isolated and identified from human teeth. Cloning forming efficiency, surface antigen expression and pluripotency were detected and identified. Enamel matrix derivatives with different concentrations (20, 50, 100 mg/L) were used to culture periodontal ligament stem cels for 2 and 4 weeks. Colagen synthesis and mineralized nodule formation were detected using Trichrom staining and Von Kosa’s staining, respectively; real-time RT-PCR was employed to detect expressions of colagen type I, osteocalcin, and RUX2; MTT and cel growth rate assay were used to detect the proliferation of periodontal ligament stem cels. RESULTS AND CONCLUSION:Periodontal ligament stem cels were spindle-shaped and showed a higher colony forming efficiency than periodontal ligament cels. The expressions of surface antigens of periodontal ligament stem cels-CD105, CD29, CD45, CD44 were respectively 99.8%, 99.7%, 1.26%, 98.8%, indicating periodontal ligament stem cels have the multilineage differentiation potential. Enamel matrix derivatives improve the colagen synthesis and mineralization nodule formation of periodontal ligament stem cels in a time-dose dependent manner. They also can improve the expression of osteogensis-related genes colagen type I, osteocalcin, RUX2 and proliferation of periodontal ligament stem cels.

Objective:To check the effect of immunosuppressants as adjuvants for DNA vaccine to prevent autoimmune disease.Methotis:Three immunosuppressants,Cyclosporin A(CsA),Tacrolimus(FK506)and Mycophenolate Mofetil(MMF)Were combined with DNA vaccine of Multiple Sclerosis(MS)and injected intramuscuarly.The splenocytes of immunized mice were checked for regulatory T cells(Treg),T cell proliferation and maturation of dendritic cells(Dcs)as well as release of intereleukin 10(IL-10).The immunized mice were induced as EAE animal model to check the preventive effect of FK506 as adjuvant.Results:More Treg cells Were induced in the mice immunized with FK506 and DNA vaccine and also T cell proliferation Was suppressed.DCs maturation Was blocked and high level of IL-10 in DCs Was induced in mice treated with FK506 and DNA vaccine.The clinic scores of EAE were significantly decreased in mice treated with FK506 and DNA vaccine compared with other control groups.Conclusion:FK506 as adjuvant of DNA vaccine can stimulate Treg cells and immune tolerance,then prevent from the autoimmune disease.

OBJECTIVE: To investigate the relationship between immune status of adenoids and secretory otitis media (SOM). METHOD: The adenoid tissue samples of 72 cases with SOM and 30 cases with adenoid hypertrophy without SOM were studied by immunohistochemical method. RESULT: The PCNA, BCL-2, CD4+, CD8+ and CD4+ / CD8+ in SOM tissue samples were 30.85 +/- 1.73, 21.27 +/- 1.25, 41.90 +/- 9.07, 20.45 +/- 7.08 and 2.10 +/- 0.17, respectively, which were much more than that of tissue samples without SOM (25.25 +/- 1.75, 14.05 +/- 1.02, 16.30 +/- 8.21, 11.15 +/- 5.71, 1.39 +/- 0.15 respectively) (P < 0.01). The expression of CD4+ in T-lymphocyte was obviously higher than that of CD8+. CONCLUSION In adenoid tissues of SOM patients, the activity of T-lymphocyte subsets are increased,the adenoids are enlarged and local immunity are enhanced. Therefore,adenoidectomy should be applied in SOM patients as early as possible.
Adenoids ; immunology ; metabolism ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Male ; Otitis Media with Effusion ; immunology ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

Objective:To study the effects of poly adenosine diphosphate ribose polymerase (PARP) inhibitors niraparib and pamiparib on the radiosensitivity of breast cancer cell lines MCF-7 and MDA-MB-436, and to explore its mechanism.Methods:MCF-7 and MDA-MB-436 cells were divided into control group, niraparib group, pamiparib group, radiation group, combination group treated with niraparib and radiation, and combination group treated with pamiparib and radiation, respectively. The effects of drugs on cell proliferation and radiosensitivity were measured by CCK-8 assay and colony formation assay, respectively. The effect of drugs combined with radiation on cell cycle and apoptosis were detected by flow cytometry. Immunofluorescence method was used to detect the changes of γ-H2AX focal number of cells. The expressions of FANCG, Bax and Bcl-2 mRNA and protein were detected by qPCR and Western blot, respectively.Results:Both niraparib and pamiparib inhibited the proliferation of breast cancer cells MCF-7 and MDA-MB-436 in a time-dose dependent manner. With the increase of irradiation dose, D0, Dq, SF2 value of MCF-7 and MDA-MB-436 cells decreased, and SER D0 and SER Dq value increased. Compared with control group, the percentages of cells in G 2/M phase were increased ( tMCF-7=41.66, 44.08, P<0.05; t436=24.69, 18.91, P<0.05), the percentage of cells in G 0/G 1 phase were decreased ( tMCF-7=8.67, 29.61, P<0.05; t436=26.39, 29.12, P<0.05), and the cell apoptosis rate was significantly increased ( tMCF-7=11.17, 11.71, P<0.05; t436=42.68, 15.89, P<0.05) in the combination group. Compared with control group, the number of γ-H2AX foci of MCF-7 cells in the radiation group and combination group treated with niraparib and radiation increased significantly at 2 h after irradiation ( t=8.89, 21.72, P<0.05). At 24 h after irradiation, the number of γ-H2AX foci basically returned to normal level in the radiation group but remained at a higher level in the combination group ( t=8.82, P<0.05). Compared with control group, the expressions of FANCG and Bcl-2 mRNA decreased ( tFANCG=14.07, P<0.05; tBcl-2=29.21, P<0.05), the expression of Bax mRNA increased ( t=8.90, P<0.05), and the expression of FANCG and Bcl-2 proteins decreased ( tFANCG=7.09, P<0.05; tBcl-2=10.24, P<0.05), while the expression of Bax protein increased ( t=2.90, P<0.05) in the combination group. Conclusions:PARP inhibitors niraparib and pamiparib can increase the radiosensitivity of breast cancer MCF-7 and MDA-MB-436 cells probably through down-regulating the expression of FANCG in FA-BRCA pathway, up-regulating apoptosis-related genes and inhibiting DNA damage repair.

Objective:To evaluate the antibacterial activity of pediatric Faropenem sodium against common pathogens isolated from children′s respiratory tract in vitro, and to provide reference for its clinical research and application. Methods:Retrospective analysis.The minimum inhibitory concentration (MIC) of Faropenem sodium, Merope-nem, Imipenem and other antibiotics was determined by the agar dilution method.A total of 156 strains of Streptococcus pneumoniae [including 32 strains of Penicillin-susceptible Streptococcus pneumoniae (PSSP), 28 strains of Penicillin-intermediate Streptococcus pneumoniae (PISP) and 96 strains of Penicillin-resistant Streptococcus pneumoniae (PRSP)], 98 strains of Haemophilus influenza, 173 strains of Klebsiella pneumoniae, and 55 strains of Moraxella catarrhali clinical isolates were used.MIC 50, MIC 90 and the accumulative inhibition of the bacteria were investigated. Results:The MIC of Faropenem sodium against all the Streptococcus pneumoniae strains ranged from 0.010-2.000 mg/L.There was no difference in the MIC distribution of Faropenem sodium against PSSP, PISP and PRSP, and the MIC 90 value was all 1.000 mg/L.Faropenem sodium inhibited all the Haemophilus influenza strains at concentrations ranging from 0.030-8.000 mg/L.There was no difference in the MIC distribution of Faropenem sodium against Haemophilus influenza with or without β-lactamase and Ampicillin resistance.The MIC 90 value was all 4.000 mg/L.Ho-wever, the MIC of Faropenem sodium against Klebsiella pneumoniae ranged from 0.250 to above 32.000 mg/L, and both MIC 50 and MIC 90 were greater than 32.000 mg/L.Faropenem sodium inhibited all the Moraxella catarrhalis strains at concentrations ranging from 0.030-2.000 mg/L, with MIC 50 being 0.500 mg/L and MIC 90 being 1.000 mg/L. Conclusions:Antimicrobial susceptibility testing results in vitro demonstrate that pediatric Faropenem sodium has satisfactory antibacterial activities against Streptococcus pneumoniae, Haemophilus influenza, and Moraxella catarrhalis, but comparatively weak antibacterial activities against Klebsiella pneumoniae.