Objective To establish a rat model of pulmonary hypertension induced by left-to-right shunt and explore the influence of high pulmonary blood flow on pulmonary vascular collagen remodeling.Methods Abdominal aorta and inferior vena cava shunting was produced in rats. Pulmonary artery meanpressure (PAMP) of each rat was measured by using a right cardiac catheterization.Pulmonary artery collagen Ⅰ and Ⅲ were detected using immunohistochemisty.Results After 11 weeks of shunting the Qp/Qs was 3.3∶1.0,indicating a large shunt. Pulmonary artery mean pressure was increased as compared with controls[(23.0?0.9) mm Hg vs (15.7? 1.1) mm Hg,P

Objective To examine the possibility that a candidate causal species of the skin and lung tumor promotion induced by exposure to dimethylarsinic acid(DMAv)and dimethylarsinous acid(DMAⅢ),caused by the induction of oxidative stress in mice.Methods Two stages lung tumotigensis animal model induced by lung tumor initiator(4-nitroquinoline 1-oxide,4NQO)and promoter(DMAv)in ddY mice,was used to examine the effect of(-)epigallocatechin gallate(EGCG)on DMAv promoting lung tumorigenesis.Two stages skin tumorigenesis animal model induced by skin tumor initiator[dimethylbenz(α)anthracene,DMBA]and promoter(DMAⅢ)in hairless mice.was used to examine the effects of DMAⅢ in skin tumorigenesis and histopathology.The goxo-2'-deoxyguanosine (8-oxodG)in lung and epidermis were analyzed by HPLC.Results The incidence of lung tumors and 8-oxodG level of lung tissue decreased significantly in 4NQO+DMAv+EGCG group.compared with 4NQO+DMAv group (0.89±0.30 vs 4.00±0.82,1.21±0.09 vs 1.53±0.32,P＜0.01).The incidence of severe keratosis in DMBA+ DMIⅢ group was more than that in DMBA group(25 vs 10,P＜0.05).An significant elevation of 8-oxodG in epidermis was observed in 0.5 h[(1.67±0.17)/105 dG],1.0 h[(1.62±012)/105 dG],2.0 h[(1.66±023)/105dG], 3.0 h[(1.60±0.15)/105 dG],compared with 0 h[(1.25±0.11)/105 dG],being significant(P＜0.05).Conclusion tumor promotion due to DMAv administration is mediated by DMAⅢ through the induction of oxidative stress.

The HPLC-MS/MS method was developed to profile the dynamics of abscisic acid (ABA) and ABA-glucose ester (ABA-GE) after cloning glycosyltransferase enzyme family gene AtUGT71C5 into Arabidopsis thaliana. By constructing over-expression lines (OE) and down-expression lines (DN), we acquired mutant strains to analyze the function of AtUGT71C5. The multiple-reaction monitoring (MRM) was used for quantitative determination in negative mode. The transition was m/z 263.1-153.0 for ABA ([M-H]t), m/z 425.1-263.0 for ABA-GE ([M-H]t), and m/z 321.0-152.0 for chloramphe-nicol. The linear range was 0.8684-217.1 ng/mL for ABA and 0.3920-196.0 ng/mL for ABA-GE. The accuracy was 88.0-109.0% for ABA and 86.6-113.0% for ABA-GE; the inter-day and intra-day precisions were less than 5.4%for ABA and 8.9%for ABA-GE, respectively. This method is simple and sensitive enough for determination of ABA and ABA-GE in A. thaliana leaves. All the evidence confirmed the speculation that AtUGT71C5 can mediate abscisic acid homeostasis.

OBJECTIVERecent studies have found that the variation of G894T on the region of T786C and 7th exon promoted by endothelial nitric oxide synthase (eNOS) gene is associated with cardiovascular disease. This research explored possible correlations between eNOS gene polymorphisms and orthostatic intolerance (OI) in children through linkage disequilibrium analysis between eNOS genes T786C and G894T and OI.

METHODSPCR, Macrorestriction Map and other molecular biotechnology were used to determine the genotypes of eNOS/T786C and G894T in 60 OI probands and their parents. Correlation analysis and transmission disequilibrium test (TDT) between T786C, G894T and OI were performed.

RESULTSThere was linkage disequilibrium of eNOS/T786C and G894T gene polymorphisms in the occurrence of childhood OI (P<0.05).

CONCLUSIONSeNOS genes T786C and G894T may be associated with the pathogenesis of OI.

Adult ; Child ; Female ; Humans ; Linkage Disequilibrium ; Male ; Nitric Oxide Synthase Type III ; genetics ; Orthostatic Intolerance ; genetics ; Polymorphism, Genetic

This study was designed to evaluate the effects of drilling through the growth plate and using adipose-derived stem cells (ADSCs) and bone morphogenetic protein-2 (BMP-2) to treat femoral head epiphyseal ischemic necrosis,which can be done in juvenile rabbits.Passage-four bromodeoxyuridine (BrdU)-labeled ADSCs were cultured,assayed with MTT to determine their viability and stained with alizarin red dye to determine their osteogenic ability.Two-month-old,healthy male rabbits (1.2 to 1.4 kg,n=45) underwent ischemic induction and were randomly divided into five groups (group A:animal model control;group B:drilling;group C:drilling & ADSCs;group D:drilling & BMP-2;and group E:drilling & ADSCs & BMP-2).Magnetic resonance imaging (MRI),X-ray imaging,hematoxylin and eosin staining and BrdU immunofluorescence detection were applied 4,6 and 10 weeks after treatment.Approximately 90％ of the ADSCs were labeled with BrdU and showed good viability and osteogenic ability.Similar results were observed in the rabbits in groups C and E at weeks 6 and 10.The animals of groups C and E demonstrated normal hip structure and improved femoral epiphyseal quotients and trabecular areas compared with those of the groups A and B (P＜0.01).Group D demonstrated improved femoral epiphyseal quotients and trabecular areas compared with those of groups A and B (P＜0.05).In summary,drilling through the growth plate combined with ADSC and BMP-2 treatments induced new bone formation and protected the femoral head epiphysis from collapsing in a juvenile rabbit model of femoral head epiphyseal ischemic necrosis.

To optimize the belt drying process conditions optimization of Gardeniae Fructus extract from Reduning injection by Box-Behnken design-response surface methodology, on the basis of single factor experiment, a three-factor and three-level Box-Behnken experimental design was employed to optimize the drying technology of Gardeniae Fructus extract from Reduning injection. With drying temperature, drying time, feeding speed as independent variables and the content of geniposide as dependent variable, the experimental data were fitted to a second order polynomial equation, establishing the mathematical relationship between the content of geniposide and respective variables. With the experimental data analyzed by Design-Expert 8. 0. 6, the optimal drying parameter was as follows: the drying temperature was 98.5 degrees C , the drying time was 89 min, the feeding speed was 99.8 r x min(-1). Three verification experiments were taked under this technology and the measured average content of geniposide was 564. 108 mg x g(-1), which was close to the model prediction: 563. 307 mg x g(-1). According to the verification test, the Gardeniae Fructus belt drying process is steady and feasible. So single factor experiments combined with response surface method (RSM) could be used to optimize the drying technology of Reduning injection Gardenia extract.
Chemistry, Pharmaceutical ; instrumentation ; methods ; Desiccation ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Gardenia ; chemistry ; Research Design ; Vacuum

OBJECTIVETo detect the expression of MAP3K5 and miR-BART22 encoded by Epstein-Barr virus and explore their relationship in nasopharyngeal carcinomas (NPCs).

METHODSFifty-three archived specimens of NPCs and 30 nasopharyngitis specimens were collected for detecting the expression of EBERs and miR-BART22 by in situ hybridization, and the expression of MAP3K5 was detected using immunohistochemistry. Ten fresh NPC and 10 fresh nasopharyngitis specimens were also obtained for determining the protein expression of MAP3K5 by Western blotting.

RESULTSEBERs were positive in all the 53 NPC specimens, and miR-BART22 was positive in 49 specimens; all the 30 nasopharyngitis specimens were negative for EBER or miR-BART22. In the 53 NPC tissues, 50 were negative for MAP3K5 expression in the cancer areas but positive in the adjacent mucosal areas, with the other 3 specimens showing a weak positivity (+). In the 30 nasopharyngitis specimens, 25 showed strong MAP3K5 positivity, 3 showed weak positivity and 2 were negative for MAP3K5 (P<0.001). Western blotting showed that the expression of MAP3K5 protein was significantly higher in nasopharyngitis than in NPC tissues (P=0.029). The expression of MAP3K5 and miR-BART22 was inversely correlated (P<0.001).

CONCLUSIONCompared with the adjacent mucosal tissues, NPC tissues have a lower expression of MAP3K5 but a higher expression of miR-BART22. The expression of MAP3K5 and miR-BART22 is inversely correlated, suggesting the possibility of MAP3K5 to serve as target gene of EBV miR-BART22. miR-BART22 may inhibit the expression of MAP3K5, thus reducing the protein phosphorylation of MAPK pathway downstream genes, inhibiting NPC cell apoptosis, preventing their differentiation and promoting their escape from immune surveillance.

Adult ; Aged ; Female ; Gene Expression Regulation, Viral ; Herpesvirus 4, Human ; genetics ; Humans ; MAP Kinase Kinase Kinase 5 ; genetics ; metabolism ; Male ; MicroRNAs ; genetics ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; virology ; Tumor Escape ; Viral Matrix Proteins ; metabolism ; Young Adult

This study aimed to examine the biocompatibility of calcium titanate (CaTiO3) coating prepared by a simplified technique in an attempt to assess the potential of CaTiO3 coating as an alternative to current implant coating materials.CaTiO3-coated titanium screws were implanted with hydroxyapatite (HA)-coated or uncoated titanium screws into medial and lateral femoral condyles of 48 New Zealand white rabbits.Imaging,histomorphometric and biomechanical analyses were employed to evaluate the osseointegration and biocompatibility 12 weeks after the implantation.Histology and scanning electron microscopy revealed that bone tissues surrounding the screws coated with CaTiO3 were fully regenerated and they were also.well integrated with the screws.An interfacial fibrous membrane layer,which was found in the HA coating group,was not noticeable between the bone tissues and CaTiO3-coated screws.X-ray imaging analysis showed in the CaTiO3 coating group,there was a dense and tight binding between implants and the bone tissues;no radiation translucent zone was found surrounding the implants as well as no detachment of the coating and femoral condyle fracture.In contrast,uncoated screws exhibited a fibrous membrane layer,as evidenced by the detection of a radiation translucent zone between the implants and the bone tissues.Additionally,biomechanical testing revealed that the binding strength of CaTiO3 coating with bone tissues was significantly higher than that of uncoated titanium screws,and was comparable to that of HA coating.The study demonstrated that CaTiO3 coating in situ to titanium screws possesses great biocompatibility and osseointegration comparable to HA coating.

BACKGROUNDAfter establishing a murine model of aminoglycoside antibiotic (AmAn) induced ototoxicity, the sensitivity of AmAn induced ototoxicity in three murine strains and the effect of kanamycin on the expression of Na-K-2Cl cotransporter-1 (NKCC1) in stria vascularis were investigated.

METHODSC57BL/6J, CBA/CaJ, NKCC1(+/-) mice (24 of each strain) were randomly divided into four experimental groups: A: kanamycin alone; B: kanamycin plus 2, 3-dihydroxybenzoate; C: 2, 3-dihydroxybenzoate alone; and D: control group. Mice were injected with kanamycin or/and 2, 3-dihydroxybenzoate twice daily for 14 days. Auditory brainstem response (ABR) was measured and morphology of cochlea delineated with succinate dehydrogenase staining. Expression of NKCC1 in stria vascularis was detected immunohistochemically.

RESULTSAll three strains in groups A and B developed significant ABR threshold shifts (P < 0.01), which were accompanied by outer hair cell loss. NKCC1 expression in stria vascularis was the weakest in group A (A cf D, P < 0.01) and the strongest in groups C and D (P < 0.05). CBA/CaJ mice had the highest sensitivity to AmAn.

CONCLUSIONSAdministration of kanamycin established AmAn induced ototoxicity. Kanamycin inhibited the expression of NKCC1 in stria vascularis. 2, 3-dihydroxybenzoate attenuated AmAn induced ototoxicity-possibly by enhancing the expression of NKCC1. Age related hearing loss did not show additional sensitivity to AmAn induced ototoxicity in murine model.

Animals ; Anti-Bacterial Agents ; toxicity ; Auditory Threshold ; drug effects ; Hair Cells, Vestibular ; drug effects ; Kanamycin ; toxicity ; Mice ; Mice, Inbred C57BL ; Mice, Inbred CBA ; Sodium-Potassium-Chloride Symporters ; analysis ; drug effects ; Solute Carrier Family 12, Member 2 ; Stria Vascularis ; chemistry ; drug effects

OBJECTIVETo establish a mice model of aminoglycoside antibiotics (AmAn) induced ototoxicity. Then to investigate the sensitivity of AmAn induced ototoxicity in three mouse strains and effect of kanamycin on the expression of Na-K-2Cl co-transporter-1 (NKCC1) in stria vascularis.

METHODSC57BL/ 6J, CBA/CaJ, NKCC1 +/- mice (each of twenty-four) were randomly divided into four experimental groups A, B, C and D (A kanamycin alone, B kanamycin plus 2, 3-dihydroxybenzoate, C 2, 3-dihydroxybenzoate alone, D control group). Mice were injected with kanamycin or/and 2, 3-dihydroxybenzoate for 14 days. Auditory function was measured by auditory brainstem response (ABR) and morphology of cochlea was observed by succinate dehydrogenase staining. Expression of NKCC1 was detected by immunohistochemistry.

RESULTSMice in group A developed significant ABR threshold shifts (P < 0.01), which were accompanied by out hair cells loss. Mice in group B significantly attenuated ABR threshold shifts with out hair cells loss (P <0.01). The immunostaining of NKCC1 in stria vascularis was attenuated significantly in group A compared with group D (P < 0.01) while the immunostaining in group B was enhanced than which in group A (P < 0.01). CBA/CaJ mice has the highest sensitivity to AmAn in three mouse strains.

CONCLUSIONSAn mouse model of AmAn induced ototoxicity could be established by administration of kanamycin. Kanamycin could inhibit the expression of NKCC1 in stria vascularis. 2, 3-dihydroxybenzoate could attenuate AmAn induced ototoxicity maybe by enhancing the expression of NKCC1. Mice that had the characteristic of presbycusis didn't show additional sensitivity of AmAn induced ototoxicity.

Animals ; Anti-Bacterial Agents ; toxicity ; Blood Vessels ; drug effects ; metabolism ; Cochlea ; blood supply ; drug effects ; metabolism ; Kanamycin ; toxicity ; Mice ; Mice, Inbred C57BL ; Mice, Inbred CBA ; Parabens ; toxicity ; Sodium-Potassium-Chloride Symporters ; metabolism ; Solute Carrier Family 12, Member 2