Objective To develop a new type of gene vaccine against plague. Methods caf1 gene of caf Operon Encoding F1 Antigen of Y. pestis was inserted into pHSS6-mTn-3xHA/lacZ library plasmids, the recombinant plasmids were linearized with Not I and transformed into yeast cell by lithium acetate (LiAc) method to induce homologous recombination. Positive recombinants were then selected with uracil-lack medium. Results These recombinants were confirmed by colony PCR to have the target gene fragments. Conclusion The present study provided a platform for constructing a novel expression system for expressing Y. pestis F1 Antigen via homologous recombination in Saccharomyces cerevisiae, which may contribute to the genetic prevention of plague through digestive-tract-route (DTR).

Objective To construct the recombinant plasmid carrying shRNA to ATX and analyze the nucle- ic acid sequence for further searching new gene therapy method of tumor.Methods Two DNA sequences containing short hairpin structure were designed and synthesized.The complement form was obtained by annealing and inserted into vector Psilcncer2.1-U6 neo,and the recombinant plasmid was transformed into DH5a strain.Finally the plasmid identified by restriction enzyme was used for sequence analysis.Results The recombinant Psileneer2.1-U6 neo car- rying shRNA to ATX had been constructed and the aim sequence had been obtained.Conclusion The construction of the recombinant plasmid carrying shRNA to ATX lays the basis for the study of its inhibitive effect on tumor.

Humans ; Imaging, Three-Dimensional ; Neoplasms ; radiotherapy ; surgery ; Radiosurgery ; instrumentation ; methods ; Tomography, X-Ray Computed

AIM:By analyzing optical coherence tomography angiography (OCTA) characteristics of central serous chorioretinopathy (CSC) and comparing the differences of CSC between OCTA and indocyanine green angiography(ICGA), to explore if OCTA can substitute ICGA for diagnosis of CSC patients, and guide the treatment of photodynamic therapy (PDT).METHODS: We reviewed 30 eyes of 30 patients with CSC, who were diagnosed by fluorescein angiography (FFA) and ICGA at Beijing Tongren Eye Center from November 2015 to March 2016.All patients underwent best-corrected visual acuity (BCVA) measurement, intraocular pressure, slit-lamp examination, indirect ophthalmoscope, color fundus photography, FFA, ICGA and OCTA.FFA and ICGA were captured by Spectralis HRA + OCT (Spectralis HRA + OCT;Heidelberg Engineering, Heidelberg, Germany).OCTA was performed by RTVue XR Avanti device (OptovueInc, Fremont, CA) with 6mm×6mm Angio Retina mode.The software (version 2017.100.0.1;OptovueInc) automatically segmented the tissue into four layers, the characteristics of choriocapillaris layer were analyzed.At the same time, the differences between OCTA and ICGA images were compared among CSC patients.The maximum diameters and areas of both choroidal hyperperfusion in ICGA and high flow signal in OCTA were measured.Then, the paired t test was used to analyze the differences between the maximum diameter and area of OCTA and ICGA measurement.RESULTS: Among 30 cases, high blood flow signals of OCTA were clearly visible in 27 cases, namely the coarse grain region;the inner low flow signals surrounded by high blood flow signals were seen in 21 cases;the outer low flow signals surrounding high blood flow signals were seen in 7 cases.High blood flow signals of OCTA were corresponded with the choroidal hyperperfusion of ICGA images;among these 30 cases, there were low reflection shadows in choroidal hyperperfusion with ICGA for 22 cases, for 21 cases out of these 22 cases, low flow signals inside of high flow signals of OCTA could be seen;9 out of 30 cases, there were low reflection halo outside of choroidal hyperperfusion of ICGA, and 7 out of these 9 cases, low flow signals outside of high flow signals of OCTA could be seen;still for those 30 cases, leakage point in late ICGA could be seen with 14 cases, however, special flow signals in OCTA could not be seen for them.For ICGA, the maximum diameter of choroidal hyperperfusion was 1.589±0.295mm, whose area was 0.705±0.131mm2;while for OCTA, the maximum diameter of high flow signal was 1.576±0.293mm, whose area was 0.745±0.138mm2.By using paired t test, there was no statistical difference between the maximum diameter of choroidal hyperperfusion in ICGA and the maximum diameter of high flow signal in OCTA, nor difference between the area of ICGA and OCTA.CONCLUSION: The high flow signals can be clearly visible in OCTA, which are corresponded with choroidal hyperperfusion in ICGA.OCTA can substitute ICGA for diagnosis of CSC patients, and guide the treatment of PDT.

Objective To explore the nervous regulating mechanism in the effect of light intensity on reproductive performance. Methods Sixty healthy Jinghong-1 hens aged 100 days were randomly divided into four groups of 15 hens and applied a light of 30 lx, 20 lx, 10 lx, and 1 lx, respectively, for 7 days. The distribution and expression of gonadotropin-releasing hormone (GnKH) in mesencephalon and diencephalon were detected using immunohistochemical and image analysis methods. Results GnKH immunoreactive positive neurons were mainly observed in stratum griseum centrale (SCC) of tectum, nucleus isthmi pars magnocellularis (Imc) , nucleus isthmi pars parvocellularis (Ipc) , nucleus subpre-tectalis/interstito-pretectalis-subpretectalis (SP/IPS), nucleus of basal optic root (nbor), and nucleus semilunaris (Slu) of mesencephalon. The mean gray values of positive neurons in 30 lx and 20 lx groups were significantly higher than those of 10 lx and 1 lx groups (P<0. 05) in SGC, Imc, Ipc, and SLu. The percent cell area in SGC, Imc, and Ipc was the largest in 30 lx group, having no significant difference comparing 20 lx, being significant higher than those of 10 lx and 1 lx (/><0. 05). GnRH immunoreactive positive neurons were mainly observed in nucleus rotundus (Rot), nucleus periven tricularis hypothalami (PHN), nucleus pariventriculairs megnocellularis (PVN) , nucleus geniculars lateralis venlralis (CLv) , nucleus dorsolaterals thalami (DLAm) , nucleus commissura pallii anterior (Cpa) , and nucleus medial septal (SM) of diencephalon. The mean gray values of positive neurons in 30 lx were the largest, being significantly higher than those of and 20 ix, 10 lx and I lx groups (P<0.05) in Rot. There was no significant difference (/'>0. 05) between 30 lx and 20 lx groups, but they were significant higher than those of 10 lx and 1 lx (P<0. 05) in PHN, PVN, and GLv. The percent cell area in Hot was the largest in 30 lx group, having no significant difference comparing 20 Ix, being significant higher than those of 10 lx and I Ix (P<0. 05). Those in 30 lx and 20 lx groups were significant higher than 10 lx and 1 lx (P<0.05) in PHN and PVN. Conclusion Light intensity affect the expression of GnRH in hen mesencephalon and diencephalon obviously in Jinghong-1 hens aged 100 days, regulating their reproductive function. The tectofugal pathway of the optical pathways, including collateral circulation of nucleus isthmi, plays an important role in the course of regulation.

Astragalus polysaccharides was lounded to 4-(2-aminoethylphenol), followed by labeling the APS-Tyr with fluorescein-5-isothiocyanate (FITC) at the secondary amino group. The absorption enhancement effects of low molecular weight chitosan and protamine on astragalus polysaccharides were evaluated via Caco-2 cell culture model. The results show that the fluorecent labeling compound has good stability and high sensitivity. On the other hand low molecular weight chitosan and protamine also can promoted absorption of the astragalus polysaccharides without any cytotoxity, and the absorption increase was more significant with increasing the amount of low molecular weight chitosan and protamine. At the same time, the low molecular weight chitosan has slightly better effect. The transepithelial electric resistance (TEER) of Caco-2 cells show that absorption enhancers could improve its membrane transport permeability by opening tight junctions between cells and increasing the cell membrane fluidity.
Absorption ; Astragalus Plant ; chemistry ; Biological Transport ; Caco-2 Cells ; Fluorescein-5-isothiocyanate ; chemistry ; Fluorescent Dyes ; chemistry ; Humans ; Plant Extracts ; chemistry ; pharmacokinetics ; Polysaccharides ; chemistry ; pharmacokinetics

Acidithiobacillus ferrooxidans are able to synthesize intra-cellular electron-dense magnetite,which makes possible that the bio-nano-magnetic particles could be obtained by cultivating Acidithiobacillus ferrooxidans. In order to isolate the strain which has the capacity to produce more magnetic particles, the solid-plate magnetophoresis method was firstly created. After isolation using the method, the rate of the cells which contain intra-cellular magnetic particles was increased from 30% to more than 90%, in addition, after isolation each cell possessed 2~5 magnetic particles which disperse in cells. The isolated cells are able to orient and migrate to the magnet in artificial magnetic field but could not orient swimming only under the geomagnetic field. Magnetosomes produced by Acidithiobacillus ferrooxidans were range from 40nm to 90 nm according to the results of TEM. Energy dispersive X-ray analysis indicated that extracted magnetic particles consisted of oxygen and iron. The results show that some Acidithiobacillus ferrooxidans cells have weak magnetotaxis and they could be able to be separated by solid-plate magnetophoresis method from others. With the development of this new isolation method, it is possible to do deeper research to generate a comprehensive description of the mechanism that how Acidithiobacillus ferrooxidans synthesize the magnetic particles.

Objective To investigate the diagnosis and treatment of idiopathicgranulomatous mastitis.MethodsThis study was to retrospectively review the clinical presentation,radiological investigation,histopathological features,treatment and outcome of idiopatbic granulomatoos mastitis of women presenting to Xuanwu Hospital between January 2002 and June 2010.ResultsTwenty-four patients with a mean age of 34.5 years presented with a diagnosis of idiopathic granulomatous mastitis.Patients presented with a palpable breast lump,breast abscess,fistula formation in different periods of the disease; the role of radiological imagings was found to be limited in differentiating idiopathic granulomatous mastitis from other inflammatory and maliguant conditions of the breast.All patients underwent a surgical procedure as the main treatment; in the form of excision or incision and drainage of the breast lesions. Mean follow-up was 47.38 ( range 6-96 ) months with recurrence in 3(12.5％) patients.ConclusionsIdiopathic granulomatous mastitis presents clinically with a palpable breast lump.The diagnosis is often only made histopathologically after surgical excision or core biopsy.Wide excision of the lesions or incision and drainage of the lesion are the main treatment modalities.

The difference between three representative components of total salvianolic acids in pharmacodynamic activity were compared by three different pharmacological experiments: HUVECs oxidative damage experiment, 4 items of blood coagulation in vitro experiment in rabbits and experimental myocardial ischemia in rats. And the effects of contribution rate of each component were calculated by multi index comprehensive evaluation method based on CRITIC weights. The contribution rates of salvianolic acid B, rosmarinic acid and Danshensu were 28.85%, 30.11%, 41.04%. Apparent oil/water partition coefficient of each representative components of total salvianolic acids in n-octyl alcohol-buffer was tested and the total salvianolic acid components were characterized based on a combination of the approach of self-defined weighting coefficient with effects of contribution rate. Apparent oil/water partition coefficient of total salvianolic acids was 0.32, 1.06, 0.89, 0.98, 0.90, 0.13, 0.02, 0.20, 0.56 when in octanol-water/pH 1.2 dilute hydrochloric acid solution/ pH 2.0, 2.5, 5.0, 5.8, 6.8, 7.4, 7.8 phosphate buffer solution. It provides a certain reference for the characterization of components.
Animals ; Benzofurans ; chemistry ; pharmacology ; Cinnamates ; chemistry ; pharmacology ; Depsides ; chemistry ; pharmacology ; Lactates ; chemistry ; pharmacology ; Male ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Solubility

Objective:To summarize our experience in perioperative anesthetic management for fulminant hepatic failure (FHF)patients receiving liver transplantation.Methods:The clinical anesthetic data of 48 FHF patients receiving orthotopic liver transplantations(OLT)from January 2006 to January 2007 were retrospectively analyzed,and the anesthetic management expe- rience was summarized.General anesthesia was applied;the hemodynamics was monitored during the operation and doses of adrenaline and phenylephrine were adjusted according to the monitoring results.Blood samples were obtained before operation, before anheptic,30 min after anhepatic phase,5 min before neohepatic phase,and 5 min,30 min and 60 min after neohepatic phase for blood gas and electrolyte analysis and for determination of coagulation function;the drugs were subsequently adjusted according to analysis results.Results:All the 48 patient underwent successful anesthetic management and there was no death dur- ing opearation.The average blood loss during operation was(5 219?478)ml.Mild alkalosis,hypokalemia,hyponatrium,and hy- pocalcemia were present before operations,pH,BE and HCO_3~- were obviously reduced 30 min after anhepatic phase and in- creased 60 min after neohepatic phase.Kalemia was obviously increased 30 min following anhepatic phase and began to increase 60 min following neohepatic phase.Calium concentration was decreased at the end of preanhepatic phase(P