Adolescent ; Child ; Child, Preschool ; Chronic Disease ; Cough ; diagnosis ; drug therapy ; etiology ; Female ; Humans ; Male ; Rhinitis ; complications ; diagnosis ; drug therapy ; Sinusitis ; complications ; diagnosis ; drug therapy

Brain ; pathology ; Cerebral Hemorrhage ; pathology ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Leukomalacia, Periventricular ; pathology ; Magnetic Resonance Imaging ; methods ; Male

To establish induction and liquid culture system for hairy roots of Danshen (Salvia miltiorrhiza), Agrobacterium rhizogenes A4, LBA9402, 15834 as test bacterium were used to infect aseptic leaves of Danshen. The hairy roots were induced and positive transgenic hairy roots were selected with PCR using rolB and rolC as the target gene. Then hairy roots of S. miltiorrhiza were harvested and salvianolic acids were extracted with 70% methanol containing 1% formic acid. The content of salvianolic acid B (SalB) and rosmarinic acid (RA) were determined by HPLC. According to the above research results, the Danshen hairy roots induced by A. rhizogenes LBA9402 were inoculated into the following group of culture media: MSOH, MS, B5, and 6,7-V liquid media. Then the same methods of extraction and determination for the content of Danshen hairy roots were adopted. Last, the hairy roots of S. miltiorrhiza induced by A. rhizogenes LBA9402 were inoculated into the MSOH liquid media with different pH values. The content of salvianolic acid were extracted with 70% methanol containing 1% formic acid and determined by HPLC. As a result, three kinds of A. rhizogenes A4, LBA9402, 15834 could induce hairy roots and Ri plasmids were integrated into the genome of S. miltiorrhiza by PCR. Danshen hairy roots induced by A. rhizogenes LBA9402 and A4 produced much more salvianolic acid, which were (3.27 ± 0.37)% [including (1.04 ±0.36)% of RA and (2.22 ± 0.29)% of SalB] and (3.17 ± 0.20)% [including (0.92 ± 0.31)% of RA and (2.25 ± 0.26)% of SalB], respectively. Hairy roots induced by A. rhizogenes LBA9402 when they were cultured in MSOH liquid media produced much more salvianolic acid, which was (4.56 ± 0.36)%, including (1.12 ± 0.26)% of RA and (3.44 ± 0.23)% of SalB. Hairy roots induced by A. rhizogenes LBA9402 produced the most salvianolic acid when they were cultured in MSOH liquid media with the pH value 4.81, which was 4.85%, including 1.16% of RA and 3.69% of SalB. So Danshen hairy roots induced by A. rhizogenes LBA9402 and A4 produced much more salvianolic acid when they were cultured in MSOH liquid media with the pH value 4.81. The research had established the foundation on genetic engineering to improve the quality of S. miltiorrhiza.
Agrobacterium ; physiology ; Benzofurans ; analysis ; metabolism ; Cell Culture Techniques ; instrumentation ; methods ; Cinnamates ; analysis ; metabolism ; Culture Media ; chemistry ; metabolism ; Depsides ; analysis ; metabolism ; Drugs, Chinese Herbal ; analysis ; metabolism ; Plant Roots ; chemistry ; growth & development ; metabolism ; microbiology ; Salvia miltiorrhiza ; chemistry ; growth & development ; metabolism ; microbiology

Objective:To investigate the possible reversal effects of 1 ,3-diphenyl-1 ,3-propanedione (DPPD)for cocaine-induced content changes of neurotransmitters of brain in mice.Methods:In this study,36 healthy ICR male mice were randomly divided into control group,cocaine group,three DPPD pretreatment groups (200,400,and 800 mg/kg)and DPPD alone group (800 mg/kg).The mice in control group were administered intragastrically with 1 % Tween 80 for 3 d,and the mice in cocaine group were administered intragastrically with 1 % Tween 80 for 2 d before cocaine was injected subcutaneously on the 3rd day.The mice in the three DPPD pretreatment groups were administered intragastrically (DPPD 200,400,and 800 mg/kg)for 3 d before cocaine was injected subcutaneously 30 min after the administration on the 3rd day.The mice in DPPD alone group were administered intragastrically with DPPD at dose of 800 mg/kg for 3 d.The mice were sacrificed 20 minutes after cocaine injection.The contents of dopamine (DA)and 5-hydroxytryptamin (5-HT)in the mice brain were determined by high performance liquid chromatography (HPLC)-fluorescence detector,the contents of glutamic acid (Glu) and γ-aminobutyric acid (GABA)in the mice brain were determined by HPLC-ultraviolet detector,and the neurotransmitter levels were compared between the groups.Results:The results showed that as com-pared with the control group,DA and GABA contents in cocaine group increased significantly (P <0.01 and P <0.05),while Glu content decreased (P <0.05).As compared with cocaine group,the DA levels in the three DPPD pretreatment groups (200,400,and 800 mg/kg)all decreased significantly (P <0.01 ).In DPPD 200 mg/kg pre-administration group,GABA content decreased (P <0.05),and the contents of the four kinds of neurotransmitters had no statistical differences with those of the control group.Conclusion:DPPD may have potential reversal effects of the content changes of neurotransmitters in mice brain induced by cocaine at a lower dose.

Background and purpose: Uveal melanoma (UM) is the most common primary intraocular malignancy in adult. Due to a high tendency for early metastasis the treatment of UM is very difficult. This study aimed to explore an effective approach for the treatment of patients with UM, we designed a strategy that combined HtrA2 gene therapy and radiation therapy. Methods:pIRES-Egr1-Omi/HtrA2 (pEgr1-HtrA2) recombinant plasmids were constructed and transfected into human UM cells (OCM-1) in vitro. The transfected cells were exposed to irradiation. HtrA2 mRNA and protein levels were detected by qRT-PCR and Western blot respectively. Assays that evaluated the apoptosis inducibility caused by HtrA2 gene therapy combined with radiation was performed by lfow cytometry. Followingly, the effects of HtrA2 overexpression on the in vitro radiosensitivity of uveal melanoma cells were investigated by clonogenic formation assay. The in vivo effects of HtrA2 gene therapy combined with radiation therapy were evaluated in different groups. Results:The recombinant plasmids could be successfully transferred into OCM-1 cells and transfection of pEgr1-HtrA2 plasmids combined with radiotherapy caused dramatically elevation of HtrA2 compared with non-irradiation cells in mRNA and protein levels, which was associated with increased apoptosis.Furthermore, we observed that the transfection of pEgr1-HtrA2 could significantly enhance radiosensitivity of OCM-1 cell in vitro. In mice bearing xenograft tumors, pEgr1-HtrA2 combined with radiation therapy signiifcantly inhibited tumor growth compared with the other treatment groups (P<0.05). Conclusion:Our ifndings indicate that radiation-inducible gene therapy may have potential to be a more effective and speciifc therapy for uveal melanoma because the therapeutic gene can be spatially or temporally controlled by exogenous radiation.

Objective To explore the influence of Azithromycin on helper T lymphocyte cell(Th)1/Th2 in peripheral blood of children with bronchitic asthma.Methods Twenty-four asthmatic children and 20 healthy children were selected.Peripheral blood mononuclear cells (PBMC) were isolated from venous blood and made into cells suspension in aseptic condition.0.2 mg/L,0.1 mg/L,0.05 mg/L and 0 mg/L Azithromycin were added into the cultures in asthmatic group.The control group was not interfered with Azithromycin.The supernatant was collected after 48 h.The levels of IFN-?,IL-4 and IL-10 in the supernatant were determined by enzyme-linked immunosobent assay(ELISA).SPSS 11.5 software was used to analyze data.Results 1.The level of IL-4 produced from PBMC of asthmatic group was significantly higher than that of control group(P0.05).2.Azithromycin 0.1 mg/L more promoted the secretion of IL-4 than the other 3 concentrations(Pa0.05).3.Azithromycin 0.2 mg/L and 0.1 mg/L more increased the level of IL-10 than the control group(P0.05).Conclusions The routine drug level of Azithromycin(0.1 mg/L) had no effects on the imbalance of Th1/Th2 of asthmatic children,but could modulate the immunological function by up-regulating the level of IL-10.

Objective To investigate the expression and role of costimulatory molecule 4-1BB on T cells of patients with rheumatoid arthritis(RA).Metheds The expression of 4-1BB on T lymphocytes from 30 RA patients and 20 healthy controls were detected by flow cytometry.Results The expression of 4-1BB on CD4~+T and CD8~+T lymphocytes from RA patients was significantly higher than that of normal control(18.56?4.08,10.33?2.13 vs 1.24?0.12,0.87?0.09,P＜0.01).There was more expression of 4-1BB on CD4~+T and CD8~+ T lymphocytes stimulated by anti-CD3 antibody from RA patients(33?4 vs 21?8,P＜0.01).In addition,the ra- tio of CD4~+T/CD8+T in RA patients was higher than that of normal controls and was positively correlated with 4-1BB~+CD4~+T cell.The expression of 4-1BB on CD4~+T in RA patients was positively correlated with the level of ESR and IgA(r=0.476,P＜0.05;r=0.659,P＜0.05).Conclusion The costimulatory molecule 4-1BB is ab- normally expressed in T lymphocytes from patients with rheumatoid arthritis.The abnormal expression of 4-1BB on T lymphocytes may play an important role in the development of RA.The expression of 4-1BB on CD4~+T cell may take part in the inflammation of RA.

Objective: To investigate the prevalence and influencing factors of adolescent depression symptoms in Zhejiang, so as to provide reference for improving their mental health. Methods: The middle school and university students in 11 cities of Zhejiang Province were selected by stratified cluster random sampling method. The depression symptoms of the adolescents were assessed by Center for Epidemiological Survey-Depression Scale ( CES-D ) and the influencing factors were analyzed by multivariate logistic regression model. Results: A total of 25 855 students were investigated, and 25 614 ( 99.07% ) valid questionnaires were collected. The detection rate of depressive symptoms was 26.86%(6 879 cases). The detection rate of depressive symptoms in girls was 29.75%, which was higher than 24.12% in boys ( P<0.05). The detection rate of depressive symptoms in high school students was 31.74%, the highest compared with other grades. The multivariate regression analysis showed that female students ( OR=1.690, 95%CI: 1.592-1.794 ), resident students ( OR=1.071, 95%CI: 1.010-1.137 ) , internet addiction ( OR=2.948, 95%CI: 2.527-3.439 ) , attempt smoking ( OR=1.516, 95%CI: 1.359-1.690 ), drinking ( OR=1.624, 95%CI: 1.525-1.729 ), bullied in the past 30 days ( OR=3.143, 95%CI: 2.938-3.363 and having serious injuries within a year ( OR=1.369, 95%CI: 1.263-1.543 ) were associated with adolescents who had depressive symptoms. Conclusions The detection rate of depressive symptoms is relative 26.86% among adolescents of Zhejiang Province. The students who are female, live on campus, have internet addiction, have been bullied or seriously injured, smoke and drink are more likely to have depressive symptoms.

OBJECTIVETo study the expression of lipoprotein related genes in subchondral bone of early experimental os-teoarthritis, which may play an important role in the pathogenesis of osteoarthritis.

METHODSAnimals are equally divided into two groups: experimental group and control group, both of which contain fifteen rats of similar weight. The right knee joints of experimental group underwent surgery,which involved in both medial collateral ligament(MCL) transaction and medial meniscectomy, while the control group was only carried out with a sham operation. Rats were killed at 1, 2 and 4 weeks postsurgery to obtain the right knee joints. Total RNA of the subchondral bone was extracted,and then hybridized to Agilent Whole Rat Genome Microarray. Differentially expressed genes analysis was used to study the chemokine signaling pathway.

RESULTSApoa5 expression was down-regulated at 1, 2 weeks post-surgery, Apoc2 expression was up-regulated at 1 week after surgery, Apol3 expression was up-regulated at 1 and down-regulated at 4 weeks post-surgery, Lrp1 expression was down-regulated at 1, 2 weeks after surgery. Lrp5 was down-regulated at 2 weeks after surgery. Gpihbp1, Lpl, Tfpi and Vldlr were up-regulated at 1 weeks after surgery. Lrpap1 and RGD1309808 were down-regulated at 4 weeks after surgery.

CONCLUSIONDysregulation of lipoprotein related genes plays an important role in pathogenesis of early experimental osteoarthritis.

Animals ; Disease Models, Animal ; Knee Joint ; metabolism ; Lipoproteins ; genetics ; Male ; Oligonucleotide Array Sequence Analysis ; Osteoarthritis ; genetics ; Rats ; Rats, Sprague-Dawley ; Transcriptome

Photodynamic therapy ( PDT ) is a new technique to diagnose and treat diseases with photodynamic effect produced by photosensitizer and light, and is now a main method of treating exudative age - related macular degeneration ( AMD ) . ln recent years, with the development of science and technology, combinations of PDT have become a research hot spot. ln this paper, we reviewed the research status of treatments on exudative AMD with PDT combined anti-VEGF drugs.