Compared with C3 plant,C4 plant had evident growth advantage,higher rates of water and nutrition using,and higher bio-yield than C3 plant,Sugarcane was one of the typical C4 plants.DNA was extracted from sugarcane leaves,primers were designed by the cDNA sequence of PPDK gene from GenBank.Then DNA was amplified by LA-PCR(Long Acute PCR) method,ligated into pMD18-T vectors,and transformed into E.coli.JM109.Full-length PPDK gene sequence of sugarcane was obtained,the sequence was 13.5kb in length.For convenient of the next experiment,two digest sites(XhoI and NotI) were introduced into primers,the full-length PPDK gene was splitted to two parts,and was ligated into pMD18-T simple vectors,respectively.Whole PPDK gene clone of sugarcane was finished.The strains were storaged in the lab.

Human ITP clinical trials have been performed based on observations of clinical uses of drugs, other than on mechanism studies in preclinical models, many studies were stopped because of unexpected complications. So there was need to develop ideal animal model of ITP to improve the understanding of the pathophysiology and the mechanisms of action of therapeutics. In this review, the history of animal models of ITP was retrospected, two modeling methods inducing passive and active models, especially transgenic rice model which can more accurately represent human disease pathophysiology, and their application in study of ITP were summarized. An animal model for human immune thrombocytopenia allows detailed studies of the mechanism, kinetics, and therapy of human immune thrombocytopenia.
Animals ; Disease Models, Animal ; Mice ; Mice, Transgenic ; Purpura, Thrombocytopenic, Idiopathic ; Rats

OBJECTIVETo investigate the Malytea Scurfpea fruit (MSF) on melanocyte adhesion and migration.

METHODHuman epidermal melanocytes were treated with MSF and Ginger respectively, then adhesion to bovine serum fibronectin-coated culture dishes was checked. Control and treated cells were also examined for migration into micropore filters coated with the same protein.

RESULTCompared with control, MSF treated melanocytes were obviously easier to adhere to the dishes and move into the filters in a dose-dependent manner. When the dose of MSF was 200 mg x L(-1), it could not reincrease melanocyte adhesion and migration. At 10 mg x L(-1), under every other concentrations of MSF, there was no marked difference among MSF-treated, Ginger-treated and untreated melanocytes (P < 0.05) when adhesion test were studied. But to migration, even at 10 mg x L(-1) MSF, there was obvious increased migration compared with MSF-untreated or Ginger-treated melanocytes (P < 0.01).

CONCLUSIONMSF has effect on melanocyte adhesion and migration, which can explain, in part, the capacity of MSF to modulate melanocyte function in vitiligo lesions.

Cell Adhesion ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Fruit ; chemistry ; Ginger ; chemistry ; Humans ; Melanocytes ; cytology ; drug effects ; Plants, Medicinal ; chemistry ; Psoralea ; chemistry

OBJECTIVETo investigate the effects of mTOR siRNA on mTOR-p70S6K signaling pathway in esophageal squamous cell carcinoma (ESCC) cells in vitro,and growth and apoptosis in transplanted tumor in nude mice.

METHODSmTOR siRNA was transfected into ESCC cell line EC9706 cells. The expressions of factors of the mTOR/p70S6K signaling pathway were detected by RT-PCR and Western blot. DNA contents and cell apoptosis were determined by flow cytometry, and cell proliferation was measured by CCK-8 assay. The effects of mTOR siRNA on the transplanted tumor growth were assessed in nude mice.

RESULTSThe levels of mTOR and p-p70S6K were significantly decreased (P < 0.05) while the level of p70S6K was increased (P < 0.05) in the cells transfected with mTOR siRNA, compared with that in untransfected cells and cells transfected with control siRNA. After being interfered by mTOR siRNA, the number of apoptotic cells was increased, cell proliferation became slower and cell cycle was arrested in G(1) phase compared with that in control cells. Also, mTOR siRNA inhibited the growth of transplanted tumor in vivo.

CONCLUSIONSmTOR siRNA can effectively interfere in mTOR-p70S6K signaling pathway, induce cell apoptosis and inhibit cell proliferation and tumor growth, suggesting that mTOR-p70S6K signaling pathway plays an important role in the carcinogenesis and development of esophageal squamous cell carcinoma.

Animals ; Apoptosis ; Carcinoma, Squamous Cell ; enzymology ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Esophageal Neoplasms ; enzymology ; pathology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; pharmacology ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; Signal Transduction ; TOR Serine-Threonine Kinases ; genetics ; metabolism ; Transfection ; Tumor Burden

OBJECTIVETo provide guidance for selection and breeding of Pesudostellaria heterophylla, agronomic traits of 3 mainly cultivated form of P. heterophylla were observed and compared in Guizhou shibing.

METHODThirteen agronomic traits of P. heterophylla from 3 cultivation form were measured and the traits were analyzed using multiple comparison, correlation analysis, multiple stepwise regression analysis and path analysis.

RESULTThe results showed that agronomic traits were different in 3 cultivation form, significant(P <0.05)or very significant correlated (P <0.01)among multiple agronomic traits, the width and the number of root, the first branch number, cleistogamous flowers, length and width of leaves, and the aboveground biomass were the main factors that affected the underground biomass, and the number of root, the aboveground biomass had a directly positive effect on the underground biomass. Meanwhile, whole length, length and width of leaves, cleistogamous flowers etc. had direct or indirect effect on the underground biomass.

CONCLUSIONAboveground biomass would be as the best indirect selection traits on breeding high yield of P. heterophylla, the first branch number, width of leaves and cleistogamous flowers world be as a better auxiliary index on breeding high variety of P. heterophylla.

Agriculture ; economics ; Animals ; China ; Hemiptera ; anatomy & histology ; growth & development ; Regression Analysis

Objective To investigate the prognostic value of acute beart block (AHB) after percutaneous transluminal septal myocardial ablation (PTSMA) in patients with hypertrophic obstructive cardiomyopathy (HOCM). Methods Ninety-four HOCM patients underwent PTSMA were included in this study. Twelve-lead electrocardiograms were obtained during and post PTSMA. Association between clinic events and incidence of post-PTSMA AHB was analyzed. Results AHB was induced in 26 patients by PTSMA and disappeared in 11 patients shortly post PTSMA, subacute intraventricular conduction disturbances was seen in 11 (42.3%), subacute I°AVB in 2(7.7% ) and subacute Ⅲ°AVB in another 2 (7.7%) patients. Among 68 patients without AHB during PTSMA, intraventricular conduction disturbances was evidenced in 14 patients (20.6% ), I°AVB in 2 (2.9% ) and Ⅲ°AVB in 1 patient (1.5%) after PTSMA. AHB patients with subacute heart block were associated with poor prognosis (conduction block duration was 42.00 h) while patients without AHB was associated with benign prognosis even with new onset of subacute heart block (conduction block duration was 7.33 h, P <0.01). Conclusion Patients with AHB during PTSMA are at higher risk for subacute heart block, especially intraventricolar conduction disturbances. AHB patients with subacute heart block were associated with poor prognosis and longer recovery time conducting system.

OBJECTIVETo explore the role of the phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) pathway in silica-induced α-SMA (α smooth muscle actin) expression in HEB (human bronchial epithelial) cell.

METHODSThe cultured HBE cells were divided into 5 groups: control, silica, PI3K inhibitor (Ly294002), both PI3K inhibitor (Ly294002) and silica at the same time and the inhibitor 24 h ahead of silica. The final concentrations of PI3K inhibitor and silica were 10 µmol/L and 100 µg/ml, respectively. Western blots were used to detect protein expressions of Akt, p-Akt, TGF-β and α-SMA. The location and expression of α-SMA were measured by immunofluorescence assay.

RESULTSHBE cell line exposed to silica can induce Akt phosphorylation, in which expressions of p-Akt were up regulated 1 times at 48 and the highest at 72 h. The expressions of TGFβ increased remarkably at 12 h and the peak at 48 h after silica exposure, while the expressions of α-SMA increased at 24 h and the highest at 72 h. However, the PI3K inhibitor (Ly294002) significantly down regulated α-SMA expression. When the cell line exposed to the PI3K inhibitor ahead of silica 24 h, the expressions of p-Akt and α-SMA were more remarkably down regulated which were decreased 1.5 times and 7.6 times respectively compare to silica exposure group. But no significant changes were found for TGFβ expressions. The immunofluorescence assay showed that silica can induce α-SMA expression, which located in cytoplasma, and PI3K inhibitor can decrease the expression.

CONCLUSIONSSilica induced α-SMA expression in HBE cell line is by targeting the PI3K/Akt pathway and PI3K inhibitor can repress α-SMA expression.

Actins ; metabolism ; Cell Line ; Chromones ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Morpholines ; pharmacology ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; Silicon Dioxide ; toxicity ; Transforming Growth Factor beta ; metabolism

To explore the feasibility of nonmyeloablative conditioning regimens, hematopoietic reconstitution, chimera level and the occurrence of GVHD after nonmyeloablative allogeneic stem cell transplantation in H-2 haploidentical mice, CB6F1 mice were used as the recipient and were divided into 3 groups, mice were pretreated five days before transplantation. Group A was pretreated with myeloablative conditioning regimens (TBI with 10.5 Gy), group B was pretreated by TBI (2 Gy) + Ara-C + Cy and group C-TBI (2 Gy) + Ara-C + CY + Flu, respectively. For all recipient mice, the prevention of GVHD was not given, and 2 x 10(7) bone marrow cells mixed 1 x 10(7) spleen cells from C57BL/6 mice were injected through tail vein on day 0, and then hematopoietic recovery, engraftment and GVHD of recipients were observed. The results of chimera detection after transplantation showed that the engraftment of group A remained full donor chimerism, and engraftments of group B and group C were associated with mixed chimerism or full donor chimerism, but the chimerism of group B remained below 80% and tended to decrease after 50 days whereas chimerism of group C was above 80% (chimerism close to or being full donor type) and preserved even after 50 days. GVHD occurred in all the recipient mice due to that prevention was not given, wherein the occurrence and death rate of GVHD in group A was obviously higher than that of group B and group C (P <0.01), but there was no statistical difference between group B and group C. In conclusion, the nonmyeloablative conditioning regimens mainly based on fludarabine can form stable and lasting engraftment in the body of recipients. The mixed chimerism established in recipients induce tolerance of transplantation and decrease or avoid the occurrence of GVHD.
Animals ; Female ; Graft vs Host Disease ; prevention & control ; H-2 Antigens ; genetics ; Haplotypes ; Hematopoiesis ; Hematopoietic Stem Cell Transplantation ; mortality ; Male ; Mice ; Mice, Inbred C57BL ; Transplantation Chimera ; Transplantation, Homologous ; Vidarabine ; analogs & derivatives ; pharmacology

Objective To study the non-fastidous antibiotic-resistance bacteria pollution in tap water from a neighborhood in Tianjin.Methods Tap water samples were collected and bacteria were isolated by R2A agar,and 16S rRNA gene sequence for the identification of bacteria isolates was used.Antibiotic susceptibility testing was performed using the Kirby-Bauer disc diffusion method.Results Among the 39 non-fastidous bacteria isolates,the resistance rate of antibioticresistance bacteria was 79.49％,including Enterococcus,Staphylococcus,Acinetobacter and Pseudomonas.Some bacteria (28.21％) showed resistance against only one antibiotic.The others (51.28％) were multiple resistant bacteria.Trimethoprim/sulfamethoxazole resistance was most prevalent (53.85％),and sulfamethoxazole resistance was also widely distributed (28.21％).Conclusion Tap water from this neighborhood is heavily polluted by non-fastidous antibioticresistantance bacteria,which deserves more attention.

In order to research the prophylactic effect of cyclosporine A (CSA) and mycophenolate mofetile (MMF) on GVHD in mice with H-2 haploidentical nonmyeloablative bone marrow transplantation, a murine model was established by using of C57BL/6J mouse as donor and (C57BL/6J x BALB/C) F(1) mouse as the recipient. The recipient mice were given CSA + MTX or CSA + MMF for prophylaxis of acute GVHD (aGVHD). The survival rate, hematopoietic recovery, and morbidity and mortality of aGVHD were observed for 50 days after transplantation. The results showed that typical aGVHD developed in the transplanted mice without prophylactic treatment during 22 to 25 days after transplantation. The morbidity of aGVHD was 75% (15/20), 40% (8/20) and 30% (6/20) and mortality was 100% (15/15), 62.5% (5/8) and 50% (3/6) respectively in unprophylactic group (control), CSA + MTX and CSA + MMF groups. In conclusion, CSA and MTX reduce the morbidity and mortality of aGVHD in mice with haploidentical nonmyeloablative bone marrow transplantation, and the effect of CSA + MMF is better than that of CSA + MTX.
Animals ; Bone Marrow Transplantation ; immunology ; Cyclosporine ; therapeutic use ; Graft vs Host Disease ; mortality ; prevention & control ; H-2 Antigens ; genetics ; Hematopoiesis ; Immunosuppressive Agents ; therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Models, Animal ; Mycophenolic Acid ; analogs & derivatives ; therapeutic use ; Transplantation Chimera