The Tagsk1 (Triticum asetium L. glycogen synthase kinase 1) gene derived from the genome of wheat salt-tolerance mutant RH8706-49 was cloned by PCR. The special primers designed according to full length cDNA sequence of Tagsk1 (AF525086). A binary expression vector pBI121-gsk1 containing Gus and Tagsk1 was constructed. And pBI121-gsk1 was introduced into the callus induced from mature embryos of salt-sensitive wheat H8706-34 and cv. China Spring by particle bombardment. The transformed callus were screened by Kanamycin and 0.5% NaCl. The salt-tolerance callus were obtained, which showed higher ability of salt-tolerance and could diffirentiate roots and buds on the medium containing 0.5% NaCl.
Adaptation, Physiological ; Biolistics ; DNA, Plant ; genetics ; Glycogen Synthase Kinases ; genetics ; Mutation ; Plant Proteins ; genetics ; Plants, Genetically Modified ; Salt-Tolerant Plants ; genetics ; Seeds ; genetics ; Sodium Chloride ; metabolism ; Transformation, Genetic ; Triticum ; enzymology ; genetics ; physiology

OBJECTIVETo experimentally investigate direct intramuscular gene transfer of nanoparticles encoding vascular endothelial growth factor for the treatment of peripheral artery disease.

METHODSThe human VEGF(165) cDNA was cloned into the eukaryotic expression vector PIRES2 under the control of cytomegalovirus promoter/enhancer. The recombinant gene was transferred into a rabbit model of chronic hindlimb ischemia by naked plasmid and nanoparticle respectively. Ischemia was induced in the hindlimb of New Zealand White rabbits by ligation of the distal external iliac artery and complete excision of the femoral artery and all its branches. At day 7 postoperation animals received VEGF(165) plasmid (10 intramuscular) or nanoparticle-VEGF(165) (8 intramuscular). With RT-PCR, immunohistochemistry analysis, and angiography, the expression and biological effects of VEGF(165) gene in experimental animals were investigated.

RESULTSTwo weeks after initiation of therapy, angiography showed that the transfer of VEGF(165) gene stimulated the formation of focal neovessels and established collateral circulation. The adductor muscle of ischemic limbs was histologically examined at day 14. Capillary density was increased among VEGF(165)-transfected rabbits, especially Nano-VEGF(165)-treated animals (Naked VEGF(165) plasmid = 50.18 per mm(2), Nano-VEGF(165) = 81.22 per mm(2), Control = 29.54 per mm(2), P < 0.05). RT-PCR showed that the transcription and expression of VEGF(165) gene in experimental group were significantly higher than those of control groups.

CONCLUSIONSIntramuscular administration of VEGF(165) induces collateral artery augmentation in the rabbit model of chronic limb ischemia. Nanoparticle can act as a vector to transfect specific gene and it will benefit gene transfer.

Angiography ; Animals ; Capsules ; Chronic Disease ; Disease Models, Animal ; Genetic Therapy ; methods ; Hindlimb ; blood supply ; Immunohistochemistry ; Ischemia ; genetics ; therapy ; Male ; Nanotechnology ; Particle Size ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; genetics ; therapeutic use

OBJECTIVEIdentify the significance of variants in papillary thyroid carcinoma (PTC), the role of extracellular matrix (ECM), MMPs, cell adhesion molecular (CAM) and cytokine in lymphatic metastasis in PTC and the value of PET-CT in diagnosis of thyroid carcinoma.

METHODSFive hundred and five cases of PTC which had complete medical records and followed up surveys were selected from the files. Clinical biological characteristic of the histological variants was investigated. Sixty cases of PTC were selected. As the important parts of micro ecosystems, ECM, MMPs, CAMs and cytokine were investigated in use of tissue chip and method of immunohistochemistry. In addition, a group of cases of thyroid carcinoma including PTC was analyzed, follicular thyroid carcinoma and medulla thyroid carcinoma (MTC) and their reports of PET-CT as the initiation.

RESULTSThe variants could be divided into three groups in terms of the rate of cervical lymph node metastasis. The high-metastases group included diffuse sclerosing variant, tall cell variant, column cell variant and diffuse follicular variant. The low-metastases group included macrofollicular variant and papillary microcarcinomas. Others are included in the moderate-metastases group. The rate of cervical lymph node metastasis of each group were 83.0% , 55.5% and 34.1% (P < 0. 05), respectively. The 10-year-survival were 75.3% , 95.8% and 100.0% , respectively. The 20-year-survival were 31.2%, 80.3% and 87.5%, respectively. The positive rate of LN, FN, MMP-2, MMP-9, TIMP-2, CD, Integrinbeta-1, ICAM-1, EGFR, TGFR-beta, VEGF-C and E-Cad in metastasis ranged from 51.6% (Integrinbeta-1) to 98.3% (CD), and that in primartumor ranged from 46.7% (FN) to 98.3% (ICAM-1). The expression of E-cad in primartumor was lower than that in normal tissues (P < 0.05). The sensitivity of PET-CT was 100% and the specificity was 85.7%.

CONCLUSIONSThe characteristic has significant difference among the variants. Individual treatment should be performed in terms of different variants. Furthermore, some molecules play an important role in the lymph node metastasis of PTC and may be considered as the focus of future study. In addition, compared with CT and Bus, PET-CT is more sensitive and has its unquestionable advantage as a whole body examination, especially for staging and detecting micro metastases, though it requires a high expense.

Adolescent ; Adult ; Aged ; Carcinoma, Papillary ; diagnostic imaging ; pathology ; Child ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Positron-Emission Tomography ; Sensitivity and Specificity ; Thyroid Neoplasms ; diagnostic imaging ; pathology ; Tomography, X-Ray Computed ; Young Adult

BACKGROUNDRecombinant human endostatin (rh-endostatin, Endostar) has been proved to be an inhibitor of angiogenesis. Docetaxel has been also considered as a common chemotherapeutic agent with inhibition of angiogenesis of malignancies. However, their function has been seldom compared and a best synergism protocol is not determined. This study aimed to compare the effects of two drugs, investigate their combined impact on human umbilical vein endothelial cells (HUVECs), a molecular basis and find ideal protocols to inhibit endothelial cell proliferation.

METHODSHUVECs on confluent growth or activated by vascular endothelial growth factor (VEGF) were treated by rh-endostatin or/and docetaxel at respective gradient concentration in following operations as cell proliferation determined by MTT assay, cell cycle distribution, apoptosis and markers of CD146, CD62E and CD105 detected by flow cytometery, the structure of the channel formed by HUVECs measured by tube formation count.

RESULTSRh-endostatin exhibited time dependent inhibition of proliferation while docetaxel showed both time and dose dependent inhibition. HUVECs accumulated in G(0)-G(1) with decreased numbers of cells in G(2) after a single treatment of rh-endostatin or that followed by docetaxel treatment. Cells accumulated in G(2) after both a single docetaxel and simultaneous administration. Both the number of cells in G(0)-G(1) and apoptotic cells were increased by docetaxel followed by rh-endostatin treatment. The number of non-apoptotic cells at G(0)-G(1) was increased by first administering rh-endostatin then docetaxel. Sequential treatment of docetaxel followed by rh-endostatin resulted in the greatest increase in apoptosis (34.7%) and the second highest apoptosis was seen with simultaneous administration (18.2%). Expression of CD146 and CD105 on confluent HUVECs was reduced at certain doses of rh-endostatin and/or docetaxel. However, rh-endostatin reduced CD105 without any apparent impact on either CD146 or CD62E expression, whereas these markers were down-regulated by docetaxel after pre-activation by VEGF. Rh-endostatin treatment maintained tube-like structures for a limited time. In contrast, docetaxel swiftly reduced tube formation. Simultaneous treatment, or docetaxel followed by rh-endostatin, exhibited a stronger inhibition on tube formation than either agent alone.

CONCLUSIONSBoth rh-endostatin and docetaxel can inhibit HUVEC proliferation while the high apoptotic rate after combined administration was probably owing to different sequent administration by docetaxel followed by rh-endostatin or simultaneous treatment. Both proliferation and adhesion molecules on HUVECs of confluent growth are down-regulated by the two drugs. The rh-endostatin decreased proliferation markers, but only slightly modified adhesion molecules, while both markers were down-regulated by docetaxel on HUVECs activated by VEGF. Rh-endostatin could maintain adhesion of HUVECs at first then induce cells apoptosis to damage tube formation. We hypothesize that it could lead to vascular normalization in short time. In contrast, docetaxel can suppress HUVEC proliferation, adhesion, and reduced tube formation swiftly due to its cytotoxicity. Combined treatments can induce a synergistic inhibition of tube formation.

Antigens, CD ; metabolism ; Apoptosis ; drug effects ; CD146 Antigen ; metabolism ; Cell Proliferation ; drug effects ; E-Selectin ; metabolism ; Endoglin ; Endostatins ; pharmacology ; Flow Cytometry ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; Humans ; Receptors, Cell Surface ; metabolism ; Recombinant Proteins ; pharmacology ; Taxoids ; pharmacology

Objective To summarize the clinical effects and experiences of rapid pore cranial drilling and ventricular drainage treatment on ventricular hemorrhage to evaluate the performance of rapid pore cranial drilling. Methods The clinical data of 3571 patients with ventricular hemorrhage performed the rapid pore cranial drilling and ventricular drainage treatment from 13 hospitals of Shandong province since 1977 were retrospectively analyzed and concluded; these data were compared with those in patients received traditional Dandy's device. Results In these 3571 patients, the cure rate was 27.1%, the improvement rate was 49.1%, and the death rate was 23.8%. Rapid pore drilling needed no scalp incision, no suction, no coagulation, or no special lighting, only needed puncturing the scalp, drilling through the cranium and dura matter, implanting drainage tube and stitching it up; one can manage it in about 5 minutes at bedside; while the traditional Dandy's drilling occupied 3 people in the operating room, needed more than 20 procedures, and plus the time transporting the patient, it needed at least 60 minutes or more to finfish the procedures. Rapid pore cranial drill device is superior to Dandy's cranial drill device in operating procedures, technical performance, operation conditions, personnel and time-consuming. Conclusion Rapid pore cranial drilling greatly simplifies the operating procedures, saves precious time for the seriously ill patients, reduces the mortality and improves the effectiveness of the treatment. After 35 years of clinical practice, to those patients seriously ill needed ventricular drainage treatment to rescue their lives, rapid pore cranial drilling is superior to traditional Dandy's drill technic, and is an effective method treating such diseases.

Objective: To elucidate the expression levels of key immune biomarkers, phosphate and tension homology deleted on chromosome ten (PTEN) and programmed cell death protein1(PD-1),of different immune tolerance pathway in classic Hodgkin's lymphoma (CHL) to further determine their clinical role and prognostic significance. Methods: The clinical features and prognostic factors of 56 CHL patients, who were admitted to the TianJin Medical University Cancer Institute from February 2003 to August 2013, were retrospectively analyzed. PTEN and PD-1 protein expression levels were analyzed by immunohistochemistry, Epstein-Barr virus encoded RNA (EBER) was performed by in situ hybridization assay. Correlations between the expression of biomarkers and clinicopathologic parameters were examined and survival analyses were performed. Results: This cohort of 56 CHL patients included 34 males and 22 females with a median age of 25 years (ranged from 7 to 71 years). In a univariate analysis, age≥45, IPS score >2, EBER positive, high expression of PTEN protein conferred inferior 5-year OS and 5-year PFS; In a multivariate model, age≥45, IPS score >2, EBER positive, high expression of PTEN protein were identified as the independent adverse prognostic factors for CHL. Conclusions: This study suggested for the first time that PTEN was independent prognostic immune biomarkers in CHL, which provided the novel therapeutic strategy of immune therapy for CHL.
Adolescent ; Adult ; Aged ; Child ; Female ; Hodgkin Disease ; Humans ; Male ; Middle Aged ; PTEN Phosphohydrolase/analysis* ; Prognosis ; Programmed Cell Death 1 Receptor/analysis* ; Retrospective Studies ; Young Adult

Objective To explore the clinicopathological features and prognosis of the patients newly diagnosed with lung adenocarcinoma with both EGFR mutation and C-MET amplification.Methods The pathological sections were reviewed.EGFR mutation was detected by amplification refractory mutation system-quantitative real-time polymerase chain reaction,and C-MET amplification by fluorescence in situ hybridization.The clinicopathological features and survival data of the patients newly diagnosed with lung adenocarcinoma with both EGFR mutation and C-MET amplification were analyzed retrospectively.Results In 11 cases of EGFR mutation combined with C-MET amplification,complex glands and solid high-grade components were observed under a microscope in 10 cases except for one case with a cell block,the tissue structure of which was difficult to be evaluated.The incidence of lung adenocarcinoma in the patients with EGFR mutation combined with C-MET amplification at clinical stage Ⅳ was higher than that in the EGFR mutation or C-MET amplification group (all P<0.001),whereas the difference was not statistically significant between the EGFR mutation group and C-MET amplification group at each clinical stage (all P>0.05).There was no significant difference in the trend of survival rate between EGFR gene group and C-MET amplification group (χ²=0.042,P=0.838),while the survival of the patients with EGFR mutation combined with C-MET amplification was worse than that of the patients with EGFR mutation (χ²=246.72,P<0.001) or C-MET amplification (χ²=236.41,P<0.001).Conclusions The patients newly diagnosed with lung adenocarcinoma with EGFR mutation plus C-MET amplification demonstrate poor histological differentiation,rapid progress,and poor prognosis.The patients are often in the advanced stage when being diagnosed with cancer.Attention should be paid to this concurrent adverse driving molecular event in clinical work.With increasing availability,the inhibitors targeting C-MET may serve as an option to benefit these patients in the near future.
Humans ; In Situ Hybridization, Fluorescence ; Retrospective Studies ; Prognosis ; Adenocarcinoma of Lung/genetics* ; Mutation ; Lung Neoplasms/genetics* ; ErbB Receptors/genetics*

Kidney stones are a common urinary system condition that can progress to kidney disease. Previous studies on the association between tea consumption and kidney stones are inconsistent. A cross-sectional study to investigate the association between tea consumption and kidney stones was conducted from 2013 to 2014 and recruited 9,078 northern Chinese adults. A total of 8,807 participants were included in the final analysis. Participants' prevalence of kidney stones was 1.07%, 1.73%, and 2.25% based on their tea consumption frequency of never, occasionally, and often groups, respectively. Compared with the 'never' group, the odds ratios (95% confidence intervals) for the occurrence of kidney stones were 1.57 (1.00-2.46) and 1.65 (1.06-2.57) in the 'occasionally' and 'often' groups, respectively. After adjusting for sex, age, and other potential confounding factors, tea consumption still significantly increased the risk of kidney stones. Tea consumption is independently associated with an increased risk of kidney stones in the investigated population, suggesting that a decrease in the consumption of tea may be a preventive strategy for kidney stones.