Objective To screen promoter DNA-binding protein of inducible nitric oxide synthase gene by using phage display technique from human liver cDNA library, and to study the expression and regulation mechanism of iNOS gene. Methods The sequence of iNOS promoter was identified in GenBank by bioinformatics based on the open reading frame(ORF) of iNOS gene and amplified from HepG2 genome by polymerase chain reaction (PCR). The amplified product was subcloned into pCAT3-Basic reporter vector, named as pCAT3-iNOSp. The HepG2 cell line was then transfected with pCAT3-Basic to serve as negative control, and pCAT3-promoter which contains the promoter region of CMV served as the positive control subject, and pCAT3-iNOSp served as the test subject, respectively. The choloraphenical acetyltransferase(CAT)activity was determined by enzyme linked immunosorbent assay(ELISA) kit. The T7 Select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaque was performed to amplify and PCR products were sequenced. Results The expression of CAT in transfection of pCAT3-PS1TP1p was 4.2 times as higher as pCAT3-Basic plasmid. Sequence analysis was performed in 12 positive plaque, which were the iNOSp binding protein. Conclusion The iNOS gene promoter identified in this study has shown to have transcription activity,and iNOS promoter DNA-binding proteins havs been screened. The results will be useful for further study of the expression and regulation mechanism of iNOS in liver cell.

Abstract?AIM: To analyze the effects of femtosecond laser assisted cataract surgery ( FLACS ) in the treatment of cataract and its effect on prognosis.?METHODS:Forty-two cases (42 eyes) of patients with cataract who were treated in the Department of Ophthalmology in our hospital between January 2012 and December 2014 were selected as the study objects. According to the order of treatment, they were divided into control group and observation group, 21 cases in each. The control group was treated with traditional phacoemulsification cataract surgery ( PCS ) . On the basis, the observation group was treated with femtosecond laser. The effective phacoemulsification time (EPT), cumulative dissipated energy (CDE), fluid flow and monitored pressure of the two groups were recorded.The rate of corneal endothelial loss and the situation of Tyndall phenomenon were statistically analyzed.The two groups were followed up for 1a.The long-term visual acuity recovery was observed.The best corrected visual acuity ( BCVA ) was recorded, and the long-term complications were statistically analyzed.?RESULTS: 1 ) The total response rate in observation group was 95% while in control group was 90% ( P>0.05);2) the surgery time of the observation group was longer than that of the control group ( P<0.05) but EPT was shorter than that of the control group.CDM and liquid flow were less than those of the control group ( P<0.05 ); 3 ) at 1d after surgery, there was no significant difference in intraocular pressure between the two groups (P>0.05); the rates of Tyndall phenomenon and corneal endothelial loss in the observation group were lower than those in the control group ( P<0.05);4) BCVA of the two groups at different time after surgery were significantly higher than that before surgery (P<0.05). However, at 1d, 3mo, 6mo and 1a after surgery, BCVA of the observation group was better than that of the control group ( P <0.05 ); 5 ) the incidence of complications in the observation group after surgery (14%) was lower than that in the control group (43%) (P<0.05).?CONCLUSION: The surgical effects of FLACS in the treatment of cataract are good.After surgery, the visual acuity of patients is improved significantly and the incidence of postoperative complications is low. However, the surgery time is long and cost is high, so it is difficult to popularize.

Air ; analysis ; Air Pollutants, Occupational ; analysis ; Humans ; Occupational Exposure ; Phthalic Acids ; analysis ; Spectrophotometry ; methods ; Workplace

Animals ; Hexanes ; blood ; toxicity ; Kidney ; drug effects ; pathology ; Liver ; drug effects ; pathology ; Lung ; drug effects ; pathology ; Male ; Rats ; Rats, Sprague-Dawley

Objective To screen 18 endophytes from Pseudolarix kaempferi Gord for molluscicidal effect and identify them by morphology. Methods Molluscicidal tests were performed according to the immersion test suggested by WHO and the strain screened was identified by the slide culture. Results The mortality rates of snails immersed by JJ18 broth salified (pH=7) were 26.7%, 76.7% and 100.0% for 24,48 h and 72 h, respectively, and 53.3% and 86.7% in 5% and 10% concentrations of JJ18 broth, respectively. The active components were extracellular moiety of the broth which had no acute toxicity to fish, and JJ18 strain belonged to Aspergillus. Conclusion Extracellular moiety of endophyte JJ18 from Pseudolarix kaempferi Gord is a new resource of molluscicide.

AIM: To gain cdcSTBX25A-fas chimeric gene bearing the regulative fragment cdcSTBX25A and opening read frame of fas in order to construct and identify eukaryotic expression vectors, pAdTrack-CMV-cdcSTBX25A-fas and pAdTrack-cdcSTBX25A-fas, which have the potential to transfer the tumor proliferative signal to promoting-apoptosis signal through up-regulate the fas expression by c-myc. METHODS: A pair of primers were designed according to the known sequences of cdcSTBX25A and fas. The cdcSTBX25A-fas chimeric gene was obtained by PCR reaction and inserted into the two plasmids pAdTrack-CMV and pAdTrack, respectively. The two recombinant plasmids were transferred into E. coli DH5?. The positive clones were screened in LB media with 50 mg/L kanamycin and identified by agarose gel electrophoresis, endonuclease digestion and PCR. The nucleotide sequence of inserted cdcSTBX25A-fas was determined by dideoxy chain termination method. Using software, BLAST was conducted to analyze the structure and sequence of the target fragments and compared with GenBank. RESULTS: The chimeric target gene, cdcSTBX25A-fas, of 1 603 bp was obtained. The positive host bacteria E. coli DH5? of recombinant plasmids were screened and amplified. The double-enzyme digestion showed the pAdTrack-CMV-cdcSTBX25A-fas and pAdTrack-cdcSTBX25A-fas presenting 9.2 kb, 1.6 kb bands, and 8.3 kb, 1.6 kb bands respectively. The sequence analysis confirmed that the two shuttle plasmids containing 1 597 bp cdcSTBX25A-fas with the ORF of fas. CONCLUSION: The eukaryotic expression plasmids pAd-Track-CMV-cdcSTBX25A-fas and pAdTrack-cdcSTBX25A-fas were successfully constructed.

AIM: To investigate the possible mechanism of PRL-2 in invasive metastasis of tumors.METHODS: The PRL-2 vector was transfected into CL1 cell with lipofectamine reagent,the transfectants were selected by growth in the medium supplemented with G418.Zymographic analysis of metalloproteinases(MMPs) activity was performed,RT-PCR was used to determine the mRNA levels of(MMP-2),MMP-9,TIMP-1 and TIMP-2,the protein levels of(MMP-2),MMP-9,TIMP-1 and TIMP-2 were analyzed by Western blotting.The effects of the special inhibitor of PRL-2 on transfected cells were also observed.RESULTS: The stable cell line selected by G418 was identified by RT-PCR and Western blotting.More abundance of MMP-2,MMP-9 and its activated type were secreted by the CL-1-PRL-2 cells than untransfected cells and transfected vector cells(P

Carotid Artery Diseases ; complications ; Fasciitis, Necrotizing ; complications ; Female ; Humans ; Middle Aged

Objective To monitor the quality changes of iodized salt and analyze its impact factor in Chongqing between 2001 and 2009. Methods Salt samples were collected according to the east, west, south,north and center locations in iodized salt production, wholesale and household sectors. Two units in iodized salt production and wholesale segment were sampled from north, south, east and west places and only 1 unit was sampled from the central place. Nine samples were collected every month in each place. If the place had less than 9 units, and then taken all the units. About resident household, 2 townships were sampled from north, south, east and west places, and 1 township was sampled from the central place, then 20 samples were collected from each township. Iodine content was detected by oxidation-reduction assay. The index of mean iodine, qualified rate from factories and wholesale, coverage rate and taking rate of qualified iodized salt in residents were calculated.Significance was analyzed by trend test, analysis of variance and X2 test. Results The qualified rate of iodized salt from the manufacturers was 92.9%(13/14) in 2001 and the rate was 100.0% each year from 2002 to 2009. The qualified rates of iodized salt from the wholesale were 88.7%(282/318) - 99.8%(431/432). The rates of 2001 and 2002 were lower than that of other years(X2 = 4.98 - 45.69, all P< 0.05 or < 0.01). The coverage rate and taking rate of qualified iodized salt in residents were 94.2% (11 154/11 841 ) - 98.9% ( 14 061/14 217), 83.5% (9 887/11 841 ) -95.8% (13 449/14 039), respectively. The rates showed an increasing tendency (F = 9.27, 26.39, all P < 0.05).The districts(counties) with qualified iodized salt consumption rate > 90% kept increasing. The mean iodine from the manufacturers and wholesale were 29.71 - 36.25, and 31.26 - 36.13 mg/kg, respectively. The iodine level showed a descending trend(F = 35.45, 140.59, all P < 0.01 ). The mean iodine level from the inhabitants were 28.84 - 30.98 mg/kg which remained stable (F = 3.05, P > 0.05 ). The iodine level from manufacturers, wholesale to inhabitants showed an descending trend(F = 38.46 - 671.23, all P < 0.01 ). Conclusions The surveillance results of iodized salt shows an increasing tendency in quality of iodized salt, eoverage rate and taking rate of qualified iodized salt. Factors that affect the quality of iodized salt is that the enterprise does not add iodine to salt strictly by the standard.

ObjectiveTo analyze the reason of cervical muscle strain misdiagnosed as cervical spondylopathy, and confirm the diagnosis and the best treatment.MethodsThe data of cervical X ray (direct, lateral and double oblique), CT and MRI (degeneration degree and lordosis of cervical spine, the changes of vertebral interspace, vertebral joint, intervertebral disc, intervertebral foramen, and transverse foramen) of 32 patients with cervical muscle strain were analyzed corresponding with clinical symptoms and signs. The attribute of the symptoms and signs were distinguished.ResultsIn all patients, besides the common symptoms and signs, there were 26 patients having degeneration and 17 alignment of the cervical spine from X ray, and 29 cases having cervical disc herniation from CT or MRI. After 5～10 times manipulation curing, 22 patients were cured, 8 patients were obviously improved and 2 patients were better than before.ConclusionCervical muscle strain and cervical spondylopathy have some similar symptoms and signs. But their attribute and therapeutics are very different. Manipulation is an effective and stable therapeutic method for cervical muscle strain.