Asthenopia ; epidemiology ; etiology ; Fatigue ; epidemiology ; etiology ; Humans ; Occupational Diseases ; epidemiology ; Railroads ; Sampling Studies

The high-copy-number plasmid pcDNA3.1+ is unstable within S almonella typhimurium. A novel plasmid pmcDNA3.1+ was constructed by removin g the promoter sequence of ampicillin resistance gene (bla gene) in plasmid pcDNA3.1+. In contrast to pcDNA3.1+, pmcDNA3.1+ was stable within Salmonel la typhimurium SL7207 in LB medium with or without ampicillin. Further experi ments showed the ?-lactamase activity of Salmonella typhimurium SL7207(pmc DNA3.1+) was apparently lowered than that of Salmonella typhimurium SL7207( pcDNA3.1+) and the high ampicillin concentration was maintained longer in LB me dium culturing Salmonella typhimurium SL7207(pmcDNA3.1+). When mice were a dministered with Salmonella typhimurium SL7207(pmcDNA3.1+) intraperitoneall y, more than 95% of Salmonella cells separated from the spleen still harbore d the plasmid pmcDNA3.1+ 7 days later; but 99% of Salmonella cells lost the plasmid pcDNA3.1+ at day 3 in mice innoculated with Salmonella typhimurium SL7207(pcDNA3.1+). By lowering the expression of bla gene, the rapid deco mposition of ampicillin in LB medium was avoided and the metabolic pressure was relieved for the host cells. This method offers a solution for the problem of t he instability of high-copy-number plasmid within Salmonella typhimurium.

AIM: To evaluate intraocular pressure (IOP) control and visual rehabilitation after placement of the Ex-press(R)miniature glaucoma shunt with adjunctive amniotic membrane transplantation (AMT) and mitomycin C (MMC) in patients with post-traumatic open-angle glaucoma during 2y of follow up.METHODS: This was an interventional,2-year,observational study.Eighteen eyes were prospectively observed (in 18 patients with traumatic secondary open-angle glaucoma) in which Ex-press miniature glaucoma filtration shunts were implanted with AMT and MMC.The outcome measures included intraocular pressure (IOP),best corrected visual acuity (BCVA),number of antiglaucoma medications,and complications.The progress of all patients was monitored for 24mo.RESULTS: Complete success (IOP <21 mmHg without glaucoma medications) was seen in 15 of the 17 (88.2%) eyes enrolled in the study at 24mo after the operation.IOP decreased from 36.9±4.8 mmHg preoperatively to 15.4±3.5 mmHg at 12mo and 15.5±3.5 mmHg at 24mo postoperatively.Early postoperative hypertension developed in two patients (11.1%) due to postoperative fibrosis.Most of the patients had improved postoperative BCVA values at the final follow-up visit compared to their preoperative measurements.Two patients (11.1%) developed transient hypotony.There were no complications such as hyphema,choroidal effusion,shallow anterior chamber,the device touching the iris,or extrusion of the device.CONCLUSION: The Ex-press miniature glaucoma filtration shunt with adjunctive AMT and MMC is effective and safe in cases of traumatic open-angle glaucoma.Surgical management is an appropriate surgical treatment in this series of cases.

Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.

OBJECTIVETo explore the changes of rat testicular spermatogenic epithelium stimulated by bacterial lipopolysaccharide (LPS) in vivo.

METHODSTwenty Wistar rats were divided into two groups: control group and experimental group. The control group was treated with pyrogen-free saline (1 ml/kg) and the experimental group was injected ip with saline containing LPS (1 mg/kg) once every two days. Two groups were operated after ten days in order to investigate the testicular pathological changes by HE staining and the expression of proliferating cell nuclear antigen( PCNA), alpha-catenin in spermatogenic epithelium by immunohistochemistry assay.

RESULTSThe testes of the experimental group showed inflammatory changes. The positive expression of PCNA in seminiferous epithelium was significantly lower than that of control group. The number of positive cells in every seminiferous, in which only spermatogonia were stained in experimental group were 59 +/- 5 and it showed significant decrease compared with the control (P < 0.01). Furthermore, the percentage of such seminiferous tubules was 0.673 +/- 0.054 and increased apparently (P < 0.01). The expression of alpha-catenin in testicular tissue of the experimental group declined (P < 0.01), and cellular positive granular light density was 0.150 +/- 0.014.

CONCLUSIONThe ability of spermatogonium proliferation and the function of conglutination of cells under inflammatory condition of the testes declined, which may be one of the etiologies of male infertility.

Animals ; Bacterial Toxins ; Disease Models, Animal ; Male ; Orchitis ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Seminiferous Epithelium ; metabolism ; alpha Catenin ; metabolism

OBJECTIVETo study effects of different interval year on Panax notoginseng plant soils middle and micro element content.

METHODThe dynamic change of Ca, Mg, Mn, Cu, Zn, B from Mabai, Matang, Gumu and Panlong were determined under different planting patterns (new soil, interval 5 years soil and continuous cropping soil).

RESULTAll the micro elements (except Ca, Mg) of interval 5 years soil and continuous cropping soil were significantly higher than those of new soil. All the middle and micro elements (except B) of interval 5 years soil were significantly higher than those of the continuous cropping soil. Planting patterns had remarkable influence on the content of Mn, Cu, B, but not Zn Ca, Mg. Cu, Ca under the 3 planting patterns, and Zn under the continuous cropping pattern did not show significant quarter changes. B content increased with the elongation of implantation time. Zn in new soil and interval 5 years also increased with prolonging of planted time. Mg, Mn and Cu content reached to peak value on April next year, and reached to minimum on the end of this experiment. Compared with new soil, the proportion of Mn, Cu in total elements increased by 29%, 114%, Mg, B decreased by 18%, 38%, Zn and Ca changed slightly of interval 5 years soils; In continuous cropping soil, Mn, Cu and B increased by 50%, 120%, 22%, respectively, but Zn, Ca, Mg had no significant change.

CONCLUSIONContinuous cropping pattern could not induce the deficient of soil middle and micro elements, and thereafter might not result in continuous cropping obstacles. But the imbalance proportional of soil middle and micro elements in P. notoginseng plant soils may be one of the main reasons for continuous cropping obstacles.

China ; Kinetics ; Panax notoginseng ; growth & development ; Soil ; chemistry ; Time Factors ; Trace Elements ; chemistry

OBJECTIVETo study effects of different interval year on Panax notoginseng plant soils macro element content.

METHODThe dynamic change of total N, P, K and available N, P, K in soil from Mabai, Matang, Gumu and Panlong was determined under different planting patterns (new soil, interval 5 years soil and continuous cropping soil).

RESULTContents order of soil total N, P and available N, P were interval 5 years soil > continuous cropping soil > new soil. No significant quarter change on soil total N was found, but the other three showed inverted "v" curve, and the peak value appeared on April 2010. Content of soil total K did not change significantly, but the available K content order was new soil > continuous cropping soil > interval 5 years soil, the quarter change was similar as soil available N or P. The soil total N, P, K and available N, P, K were different of the 4 monitoring sites under the 3 interval planting modes. There was a significant correlation between soil total P and available P under all these 3 interval planting modes, but N and K. The propitiation of N-P-K of new soil, interval 5 years soil and continuous cropping soil were 1: 0.4: 2.4, 1: 0.4:1.4, 1:0. 4:2.0, respectively.

CONCLUSIONContinuous cropping pattern induce the accumulation of P, but deficient of K. The imbalance proportion of N, P and K was one of the incentives of continuous cropping induced obstacles. Strengthen the research of optimum proportion of soil N, P and K, and then eliminate continuous cropping obstacles by means of formulated fertilization is the future research direction.

Agriculture ; methods ; Breeding ; China ; Drugs, Chinese Herbal ; analysis ; Fertilizers ; analysis ; Nitrogen ; analysis ; metabolism ; Panax notoginseng ; chemistry ; growth & development ; metabolism ; Phosphorus ; analysis ; metabolism ; Potassium ; analysis ; metabolism ; Quality Control ; Soil ; chemistry

The yeast strain (Y18) was isolated from a soil sample collected from Fildes Peninsula, Antarctica. The strain is a psychrophilic yeast with optimum and maximum growth temperatures of 10 degrees C and 18 degrees C, respectively. Teliospores were formed after 7 d on malt agar, when the germination of teliospores was observed. Both inositol and D-glucuronate were assimilated. Positive results of the DBB (diazonium blue B) color reaction, urease test, and starch formation were observed. The major CoQ is Q(8). All results indicated that Y18 belongs to the genes of Mrakia. The 18S rDNA sequence analyses showed that Y18 is closely related to Mrakia frigida. DNA-DNA relatedness study, and some biochemistry characteristics indicated that Y18 represents a new species for which Mrakia psychrophila sp. nov. is proposed.
Antarctic Regions ; Basidiomycota ; classification ; genetics ; China ; DNA, Fungal ; genetics ; Phylogeny ; Soil Microbiology

Objective To investigate the clinical and mammographic features of plasma cell mastitis.Methods Twenty-five patients(28 lesions)with histologically confirmed plasma cell mastitis, aged from 26 to 70 years(mean age 41 years),were examined with X-ray mammography.The clinical manifestations and imaging features were retrospectively reviewed.Results No case was in lactation.The painful irregular masses,ranged from 1.3 to 8cm in size,were found in 22 patients,while 3 patients with acute episode.Recurrent episodes of breast masses were noted in 4 patients.Based on the mammographic appearances,the plasma cell mastitis were classified as the following four types:inflammation-like type (2/28),ductal ectasia type(3/28),focal infiltration type(10/28)and nodular type(13/28).The valuable radiogyaphic signs:(1)An asymmetrically increased density along the lactiferous duct with a flame-like appearance,inhomogeneous low density tubular structures and scattered stick-shape calcifications.(2) Architectural distortion and oil cysts formation in adjacent area,(3)Subareolar ductal ectasia.Conclusions The clinical and mammographic characteristics of plasma cell mastitis are critical to avoiding unnecessary surgery.Histopathological result is needed for the diagnosis in patients highly suspected of malignancy.

Objective To investigate the effects of hypoxia on the features and chemoresistance of cancer stem cells in Hep-2 cells and underlying mechanism. Methods The shRNA interference recombinant plasmid targeting HIF-1α was synthesized and transfected into Hep-2 cells. The HIF-1αknockdown Hep-2 cells were established after clonal selection and the expression of HIF-1α was measured.The cellular features including proliferation,clonal formation,cell cycle,apoptosis and CD133 phenotype were measured in Hep-2 cells cultured under hypoxic condition in vitro. CD133 + cells were sorted from Hep-2 cells with flow cytometry. Clonal formation test and cisplatin treatment were carried out,and theexpressions of related genes( Oct-4,suvivin and p53 ) in CD133 + cells were measured.Results HIF-1αknockdown Hep-2 cells was successfully established,as evidenced by the reduced mRNA and proteinexpressions of HIF-lα. The Hep-2 cells cultured under hypoxic microenvironment showed higher proliferation and clonal formation activity,cell cycle arrest in G0/G1,lower apoptosis,up-regulated CD133,however the effects of hypoxia reduced in HIF-1α knockdown Hep-2 cells.CD133 + cells were successfullysorted from Hep-2 cells,and the CD133+ cells showed increased clonal formation activity and cisplatintreatment resistance in hypoxia. Also the effects of hypoxia on CD133 + cells decreased with HIF-1αknockdown,showing down-regulated Oct-4 and survivin and up-regulated p53.Conclusions Hypoixa caninduce the features of cancer stem cells in Hep-2 cells and increase proliferation,differentiation and chemoresistant ability of CD133 + cells,which might be correlated with the changes in expressions of HIF-1 αand related genes regulated by HIF-1 α.