In a previous study, we cloned popW from Ralstonia solanacearum strain ZJ3721, coding PopW, a new harpin protein. The procaryotically expressed PopW can induce resistance to Tobacco mosaic virus (TMV), enhance growth and improve quality of tobacco, when sprayed onto tobacco leaves. Here, we constructed an expression vector pB- popW by cloning popW into the bionary vector pBI121 and transformed it into Agrobacterium tumefaciens strain EHA105 via freeze-thaw method. Tobacco (Nicotiana tobacum cv. Xanthi nc.) transformation was conducted by infection of tobacco leaf discs with recombinant A. tumefaciens. After screening on MS medium containing kanamycin, PCR and RT-PCR analysis, 21 T3 lines were identified as positive transgenic. Genomic intergration and expression of the transferred gene were determined by PCR and RT-PCR. And GUS staining analysis indicated that the protein expressed in transgenic tobacco was bioactive and exhibited different expression levels among lines. Disease bioassays showed that the transgenic tobacco had enhanced resistance to TMV with biocontrol efficiency up to 54.25%. Transgenic tobacco also exhibited enhanced plant growth, the root length of 15 d old seedlings was 1.7 times longer than that of wild type tobacco. 60 d after transplanting to pots, the height, fresh weight and dry weight of transgenic tobacco were 1.4, 1.7, 1.8 times larger than that of wild type tobacco, respectively.
Agrobacterium tumefaciens ; Animals ; Bacterial Outer Membrane Proteins ; genetics ; Disease Resistance ; genetics ; Genetic Vectors ; Phenotype ; Plant Diseases ; prevention & control ; virology ; Plants, Genetically Modified ; genetics ; Tobacco ; genetics ; Tobacco Mosaic Virus ; Transformation, Genetic

OBJECTIVETo observe the vasodilating effect of adrenomedullin 2 (ADM2) by antagonizing angiotensin 1 (Ang II), and to explore its mechanism.

METHODSEighteen male, 180-200 g SD rats were randomly divided into 3 groups (n = 6): control group, Ang II (150 ng/(kg x min)) group and Ang II (150 ng/(kg x min)) + ADM2(500 ng/(kg x h)) group. Mini-osmotic pumps filled with peptide were implanted in the back of rats subcutaneously. After two weeks, the blood pressure was measured by the way of carotid intubation. The plasma was collected for the detection of nitric oxide (NO) content and the activity of endothelial nitric oxide synthase (eNOS). The in situ oxidation of fluorescent dye dihydroethidium (DHE) was used for detecting superoxide in rat arteries. The rat isolated arterial rings were made for studying the vasodilating effect of ADM2. Human umbilical vein endothelial cell line EA. hy 926 cells were cultured and their intracellular reactive oxygen species (ROS) were evaluated by probe DCFH-DA.

RESULTSADM2 dramatically decreased the blood pressure in angiotensin II-induced hypertension rat model, enhanced plasma NO content and the activity of eNOS and reduced superoxide formation in vessel walls. ADM2 also induced relaxation of the vascular rings preconstricted by Ang II in a concentration-dependent and endothelium-dependent manner. In cultured vascular endothelium, ADM2 ameliorated the ROS generation induced by Ang II.

CONCLUSIONAdrenomedullin 2 relaxed blood vessels by antagonizing angiotensin II-induced oxidative stress and improving the vascular endothelial function.

Adrenomedullin ; pharmacology ; Angiotensin II ; pharmacology ; Animals ; Antihypertensive Agents ; pharmacology ; Blood Pressure ; drug effects ; Drug Antagonism ; Endothelium, Vascular ; drug effects ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Male ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type III ; blood ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Vasodilation ; drug effects

Animals ; Animals, Newborn ; Cells, Cultured ; Fibroblasts ; cytology ; metabolism ; Myocardium ; cytology ; Pravastatin ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; genetics ; metabolism

OBJECTIVETo observe the difference in therapeutic effect of Chinese herbal granule decocted separately and mingly.

METHODSOne hundred patients of functional dyspepsia of Spleen deficiency and Liver stagnancy type were treated with Jianweishu Granule (JWSG, a self-formulated recipe by the author). Half of the patients received JWSG decocted separately, half of them received that decocted mingly. The therapeutic effects between the two groups were compared. Experimental observation on the effects of differently decocted remedies in rats was also conducted.

RESULTSThe therapeutic effects obtained in the two groups were similar, with respective cure rate of 72% and 70% and the total effective rate 96% in both group. There was insignificant difference between the two groups (P > 0.05). Experimental study all showed no difference between the differently decocted remedies in inhibiting gastric acid, pepsin activity, gastric function regulation, small intestine movement improvement and pain alleviation in rats.

CONCLUSIONThe therapeutic effects of JWSG decocted separately or mingly are the same. This fact provides a scientific basis for clinical use of granule form of single Chinese herbs.

Adult ; Animals ; Diagnosis, Differential ; Drug Combinations ; Drug Interactions ; Drugs, Chinese Herbal ; administration & dosage ; Dyspepsia ; drug therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Mice ; Middle Aged ; Phytotherapy ; Random Allocation ; Rats ; Rats, Wistar ; Stomach Ulcer ; drug therapy ; Yang Deficiency ; drug therapy

[ ABSTRACT] AIM:To evaluate the effects of 1α, 25-dihydroxyvitamin D3 on T helper cell 17 ( Th17 cells) and its related cytokines in a mouse model of corneal allograft transplantation.METHODS:C57BL/6 mice were transplanted with corneal grafts from BALB/c mice and treated intraperitoneally with 1.0μg 1α, 25-dihydroxyvitamin D3 or soybean oil every other day after operation.The transparency of the corneal grafts was evaluated for potential rejection signs by slit lamp biomicroscopy and histopathology.The expression levels of IL-17, RORγt and IFN-γin the spleen were measured by real-time PCR.Moreover, the protein expression of RORγt and IL-17 in the peripheral blood was analyzed by Western blotting. IL-17 and IFN-γin peripheral blood were measured by flow cytometry.RESULTS:1α, 25-dihydroxyvitamin D3 signifi-cantly inhibited the rejection of the corneal allograft and reduced the numbers of inflammatory infiltrates in the corneal graft. In the spleen, 1α, 25-dihydroxyvitamin D3 treatment reduced the expression levels of IL-17, RORγt and IFN-γ.In the pe-ripheral blood, 1α, 25-dihydroxyvitamin D3 treatment downregulated the expression levels of RORγt, IL-17 and IFN-γ. CONCLUSION:The effects of 1α, 25-dihydroxyvitamin D3 on suppressing corneal transplantation-induced allograft rejec-tion in mice are closely associated with its modulation on IL-17 and related cytokine RORγt.

Objective:To evaluate the rapid nucleic acid amplification detection of Mycoplasma pneumoniae (MP)-DNA and MP-RNA in the diagnosis of MP infection and therapeutic values in children. Methods:Patients who were diagnosed with pneumonia were enrolled from the Department of Respiration, Children′s Hospital of Capital Institute of Pediatrics from January 2018 to December 2018.Specimens were detected using the MP and Macrolide-Resistant isolates Diagnostic Kit (PCR Fluorescence Probing, Jiangsu Mole Bioscience Co., Ltd.) and MP Diagnostic Kit (Isothermal RNA amplification, Shanghai Rendu Biotechnology Co., Ltd.).Results:Among them, 42.1%(840 cases) of the 1 994 cases were positive for MP-DNA, and the macrolide associated gene mutations were detected in 96.0% (806/840 cases) of them, while 33.9% (551 cases) of 1 624 cases were positive for MP-RNA.Seven hundred and fifty-eight specimens were simultaneously detected by adopting MP-DNA and MP-RNA, and the positive rate was 43.1% (327/758 cases) and 36.7% (278/758 cases), accordingly, which were inconsistent (Kappa=0.604) in 613 (80.9%, 613/758 cases) cases, with significant differences ( χ2=6.60, P=0.01). Part of the specimens were rechecked with the interval of 7 days: MP-RNA was negative in 70.1% (47/67 cases) specimens and MP-DNA was negative in 36.1% (22/91 cases) specimens ( χ2=33.20, P<0.01). Conclusions:The positive detection rate of MP was at a high level in 2018, in Beijing, China.The results of MP-DNA and MP-RNA are consistant.But RNA detection can help to diagnose MP in the early stage, and monitor the survival of MP and its efficiency.

Background Interleukin (IL)-1a-889C/T,IL-1β-S11C/T,+ 3962C/T and IL-1Ra-2 are different morphology of single nucleotide of interleuki-1 (IL-1).The potential relationship of IL-1 polymorphisms with Beh(c)et disease has been studied in several populations and groups.However,study outcomes are controversial for a long duration.Objective This Meta-analysis was to determine whether the IL-1 gene polymorphisms is associated with the pathogenesis and development of Beh(c)et disease.Methods The literature of the assoiation between IL-1 polymorphisms and Beh(c)et disease was retrieved from Medline,EMBASE,Cochrane Library,Web of knowledge,Google Scholar,Wanfang and CNKI databases.All the published original articles (case-control studies) were reviewed with the deadline May 31,2013,and the languages were limited to English and Chinese.Included reports were evaluated based on Newcastle-Ottawa Scale (NOS) score.The potential influence of IL-1α-889TT gene,IL-1β-3962C,IL-1β-511T and IL-1Ra-2 polymorphisms on Beh(c)et disease were analyzed.RevMan5.0,the Cochrane collaboration software program,was used to prepare and complete this review.The effect size was assessed using fixed effect model in the index with lower heterogeneity (I2 ＜50％) or randomized effect model in the index with higher heterogeneity (I2＞50％).Results The literature search resulted a total of 370 cases-controlled studies,and 7 studies met the included criteria,with the NOS scores ≥ 8.The total patients were 499 and normal controls were 708.Meta-analysis was performed on several populations.Overall,the polymorphisms of IL-1β-3962C allele increased the susciptibility of Beh(c)et disease (OR =1.41,95％ CI:1.06-1.88,P =0.02),and IL-1α-889TT genotype reduced the risk of Beh(c)et disease (OR =0.61,95 ％ CI:0.40-0.92,P =0.02).However,There was no significant association between variants of IL-1β-511T allele (OR =0.84,95％ CI:0.58-1.23,P=0.38) or IL-1RA-2 allele (OR =1.25,95％ CI:0.50-3.14,P=0.63) and Beh(c)et disease susciptibility.Conclusions These results suggest that Beh(c)et disease is associated with the IL-1 gene polymorphisms at the locations α-889C/T and β-3962C/T.A larger sample size clinical data still are need to confirm this conclusion.

Objective To determine the effects and dosage of N?acetylcysteine( NAC) in the im?provement of the visibility in esophagogastroduodenoscopy( EGD) . Methods A prospective randomized con?trolled study was performed on 193 patients scheduled for EGD from November 2014 to July 2015 were ran?domized into five groups using digital table. In group A, 100 mg dimethicone and 2 g NaHCO3 were given. In group B,100 mg dimethicone, 2 g NaHCO3 and 20 000 U pronase were given. Group C received 100 mg dimethicone, 2 g NaHCO3 and 200 mg NAC. Group D received 100 mg dimethicone, 2 g NaHCO3 and 400 mg NAC and group E 100 mg dimethicone, 2 g NaHCO3 and 600 mg NAC.The agents were dissolved in 100 ml water for each patient.Endoscopy was completed by one endoscopist and the score of image visibility assessment was completed by 2 other endoscopists. The 3 endoscopists were unaware of grouping. The total scores, the time of washing, the time of examination and complications were compared and analysed. The total image scores of group A, B, C,D and E were 30?83±3?78, 35?69±2?88, 33?16±3?90, 34?95±3?46 and 36?76±2?91, respectively. Group A was the lowest(P<0?05),followed by group C(P<0?05). There were no differences among group B,D, and E(P>0?05).Images that were scored 3 were the most in group E.The washing times of each group were 38?00±19?10, 17?03±11?44, 15?92±10?81, 15?78 ±10?24 and 15?55±9?69, and the examination times of each group were 13?49±2?49, 9?41±1?86, 9?08± 1?80, 8?73±1?91 and 8?78±1?79 minutes.Group A was the longest in these two indices(P<0?05). There were no significant differences among group B, C, D and E ( P<0?05) . There were no significant differences in adverse effects among groups after EGD( P>0?05) . Conclusion The preoperative NAC can improve the visibility in EGD. The best dose is 600 mg, whose effects and safety were similar to those of 20 000 U, but yield to less washing time and the examination time in EGD.

OBJECTIVETo prove the existence of functional area of Back-shu and provide a new thinking for clinical practice.

METHODSThe consistency between the Back-shu points and Jiaji points at the same level of spinal column was discussed from the origin and development, mechanisms and clinical application of Back-shu and Jiaji points.

CONCLUSIONThe points at the same level of spinal column has same origin of development, mechanisms and clinical functions, so as to prove the existence of functional area of Back-shu.

Acupuncture Points ; Humans

Objective ， To investigate the role of serum chemokines and oxidative and antioxidant biomarkers in occupational （ silicosis） Methods silicosis hereinafter referred to as . A total of 58 patients with stage Ⅰ silicosis were selected as the - （ ）， research subjects using convenient sampling method. The serum levels of nuclear factor erythroid 2 related factor 2 Nrf2 -（ - ） - （ - - ） - heme oxygenase 1 HO 1 and 8 isoprstaglandin F2α 8 iso PGF2α were determined by enzyme linked immunesorbent assay. （ ） （ - ） The serum levels of lipid peroxide LPO and total antioxidant capacity TAOC were determined by chemistry colorimetric method. - - （ - ）， Luminex flow fluorescence technology was used to detect the serum levels of interferon γ inducible protein10 IP10 macrophage （ ）- ， - - （ ） inflammatory protein MIP 1α MIP1β and macrophagederived chemokine MDC . The above indicators were analyzed by factor Results - analysis. The information extraction rate of the original indicators of the nine biomarkers was 58.5%96.5%. Four common ， ， （ ） ， factors were extracted including Nrf2 antioxidant signaling pathway helper T cell Th 1 dominant chemotaxis the total ， ， ， ， ， oxidation/antioxidant balance and Th2 dominant chemotaxis whose variance contribution rates were 32.2% 19.1% 16.4% ， ， Conclusion - and 11.8% respectively and the cumulative variance contribution rate was 79.5%. Both the oxidant antioxidant ， disturbance and the dominance chemotaxis are involved in the occurrence and development of silicosis and the Nrf2 antioxidant signaling pathway plays the most critical role.