Adenocarcinoma, Papillary ; blood supply ; metabolism ; pathology ; Adult ; Aged ; Carcinoma, Pancreatic Ductal ; blood supply ; metabolism ; pathology ; Chemokine CXCL12 ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymph Nodes ; metabolism ; Lymphatic Metastasis ; Male ; Microvessels ; pathology ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; metabolism ; pathology ; Pancreas ; metabolism ; Pancreatic Neoplasms ; blood supply ; metabolism ; pathology ; Receptors, CXCR4 ; metabolism

Background Light-induced retinal damage models vary as many influence factors,herein the modeling method is difficult to copy.It is necessary to establish the grading standardization of retinal damage after retinal light exposure.Objective This study was to improve the modeling method and establish a grading standardization for light-induced retinal damage in rat.Methods Twenty-four SPF 8-10 week-old male SD rats were randomly divided into 4 groups and 6 eyes for each group.The rats were exposed to light intense of 5000 lx for 1,2,3 hours respectively in 3 groups,and other 6 rats served as the normal group.Full-field light exposure experiment was performed for each individual rat separately,and an annular illumination box was used tO ensure the experimental rat moving in a single direction and exposing the right eye in 5000 lx light surrounding during experimental duration.Ganzfeid electroretinogram(ERG)was recorded from the experimental rats at the fifth day after light exposure,and the animals were then sacrificed for histopathology observation to evaluate the retinal thickness change.All procedures which involved animals adhered to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.Results After exposing to intensity light for 1,2,3 hours,the b-wave amplitudes of rod response,maximal mixed response,oscillatory potential in scotopic ERG as well as cone response,20 Hz flicker response of photopic ERG were significant declined as lapse of light exposure time(F=71.690,P=0.000；F=56.250,P=0.000；F=23.610,P=0.000：F=27.130,P=0.000；F=27.030,P=0.000)and lowed by 26.2％,52.5％,70.7％,24.4％,39.3％,58.1％respectively at the end of experiment.Meanwhile,the b-wave latencies of rod response,maximal mixed response in scotopic ERG as well as cone response of photopic ERG were evidently different among different groups (F=1.370,P=0.282；F：0.800,P=0.508；F=11.840,P=0.000；F=2.080,P=0.136).Light induced retinal damage located mainly at the temporal retina area.After intensity light exposure for 1,2,3 hours,the thickness of outer nuclear layer at the superior temporal retina attenuated by 11.3％,25.6％and 72.5％,respectively(P<0.05).A significant difference was seen in mean thickness of outer nuclear layer at superior temporal retina among different groups(F=410.27,P=0.000). Conclusion A standardized grading method for light-induced retinal damage is recommended.The continuous illumination in a intensity of 5000 Ix for 1,2,3 hours can induce the mild,moderate or severe retinal damage respectively at temporal retina.

OBJECTIVETo obtain highly purified tetanus toxin fragment C (TTC) with good immunogenicity.

METHODSThe gene fragment encoding TTC was amplified from Clostridium tetani plasmid DNA by PCR, inserted into the vector pET43.1a (+) and expressed in E. coli BL21(DE3)plysS. After purification using Ni2+-chelate affinity chromatography, the expressed fusion protein was digested by thrombin and the resultant TTC protein was purified with Ni2+-chelate affinity chromatography followed by identification with SDS-PAGE and Western blotting. The purifed TTC protein was then used to immunize mice to test its immunogenecity.

RESULTSThe 1373-bp gene fragment encoding TTC was obtained, and the constructed recombinant expression vector pET43.1a (+)-TTC was successfully expressed in E. coli BL21(DE3)plysS. SDS-PAGE identified a recombinant fusion protein with relative molecular mass (Mr) of 117 000, which accounted for 22% of the total bacterial protein. The TTC protein with Mr of 50 000 was obtained after purification of the thrombin digestion products of the fusion protein, with a purity reaching 95.5%. Both the fusion protein and TTC protein could be recognized by anti-tetanus toxin antibody as shown by Western blotting. The titer of the anti-serum from mice immunized with the TTC protein was 1:25 600, and the anti-serum could specifically bind to tetanus toxin.

CONCLUSIONHighly purified and immunogenetic TTC protein has been successfully obtained, which provides a good model antigen for studying antigen presentation and immune responses in vivo.

Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Peptide Fragments ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Tetanus Toxin ; biosynthesis ; genetics ; immunology ; Tetanus Toxoid ; immunology

OBJECTIVETo clone the gene coding for the peptidoglycan recognition protein (PGRP) domain (PGRPd) of mouse long PGRP (mPGRP-L) and express the protein in E. coli.

METHODSThe cDNA fragment encoding PGRPd of mPGRP-L was obtained by RT-PCR from the total RNA of Balb/C mouse liver cells and cloned into pUCm-T vector. The recombinant plasmid were identified by PCR, restriction endonucleases and sequence analysis. The PGRPd gene fragment was amplified by PCR from the recombinant plasmid, inserted into pQE-30 vector and transformed into E. coli strain M15, and the expressed PGRPd protein was purified.

RESULTSA cDNA fragment of about 500 bp was amplified by RT-PCR and the recombinant plasmid, pmPGRPd, was constructed by linking the fragment to pUCm-T vector. The results of restriction mapping of the recombinant vector were consistent with those of computer analyses. Sequence analysis showed that the cloned gene fragment (518 bp) had identical sequence with the gene encoding PGRPd of mPGRP-L gene in GenBank. The recombinant expression vector pQE-PGRPd was constructed and expressed in E. coli M15. SDS-PAGE showed that the expressed product existed mainly in the lysate supernatant as a soluble protein with relative molecular mass of 29 kD.

CONCLUSIONThe PGRPd cDNA of mPGRP-L has been successfully cloned and expressed in E. coli, which provides the basis for further study of PGRP molecule.

Animals ; Base Sequence ; Carrier Proteins ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Female ; Liver ; chemistry ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Plasmids ; genetics ; Prokaryotic Cells ; metabolism ; Protein Structure, Tertiary ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVETo express the carbohydrate recognition domain (CRD) of Balb/C mouse mannan binding lectin A (MBL-A) in E.coli.

METHODSThe target gene fragment was obtained by PCR from the plasmid pmMBL-A harboring mouse MBL-A gene. The PCR product was recombined with the prokaryotic expression vector pET-41a(+) and the resulting recombinant plasmid was identified by PCR, restriction analysis and sequencing before transformation into E.coli BL21(DE3) cell for expression of the target protein. After washing and renaturation, the protein was purified on GST-Tag purification resins and analyzed by SDS-PAGE, Western blotting and enzyme-linked immunosorbent assay (ELISA).

RESULTSA DNA fragment of about 450 bp was amplified by PCR and the recombinant plasmid pET41a-mMBL-A-CRD was constructed by linking the fragment with pET41a(+) vector. The result of restriction enzyme analysis and sequencing of the selected clones were consistent with those by computer analysis. The recombinant vector was expressed in E.coli BL21(DE3), and the expressed protein existed mainly as inclusion bodies, whose relative molecular mass was about 47,000 by SDS-PAGE analysis. After washing, renaturation and purification, the purity of recombinant protein was about 90%. Western blotting suggested immunoreactivity of the purified protein with anti-GST antibody, and its sugar binding activity was verified by ELISA.

CONCLUSIONWe have successfully obtained mouse MBL-A CRD protein, which provides the base for further functional study of the MBL-A molecule.

Animals ; Carbohydrates ; chemistry ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Inclusion Bodies ; metabolism ; Mannose-Binding Lectin ; biosynthesis ; chemistry ; genetics ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; genetics

Objective To evaluate left ventricular function in patients with hypertensive hypertrophic cardiomyopathy(HHC)using real-time 3-dimensional echocardiography(RT-3DE).Methods Thirty patients with HHC and 32 control subjects were studied.Full-volume RT-3DE data from apical window were acquired,and regional volumetric time curves of 17 segments were obtained by fast 3-dimensional border detection software.Several left ventricular function parameters were calculated semiautomatically,including global left ventricular end-diastolic volume(EDV),end-systolic volume(ESV),left ventricular ejection fraction(LVEF),the ratio of ESV/EDV of 17 segments,the standard deviation(SD)and difference(Dif)(adjusted by the R-R interval) of time to minimum systolic volume(Tmsv)in 16 segments(Tmsv16-SD and Tmsv16-Dif).Results EDV and ESV were significantly larger in patients with HHC than that in control subjects[(88±29)ml vs (72±15) ml,t=-2.680,P=0.008;(28±10)ml vs (22±6 )ml,t=-2.613,P=0.01].HHC had a higher ratio of ESV/EDV at interventricular septum(IVS)compared with control group[mid-segments of anterior IVS:(40.51±20.28)% vs (26.43±10.10)%,t=-3.378,P=0.002;mid-segments of posterior IVS:(41.44±23.55)% vs (24.46±8.12)%,t=-3.688,P=0.001;apical segments of IVS:(30.96±21.31)% vs (19.53±7.33)%,t=-2.745,P=0.01].In patients with HHC,Tmsv16-SD and Tmsv16-Dif were significantly longer[(2.48±1.38)% vs (1.16±0.26)%,t=-5.117,P＜0.001;(7.67±5.07)% vs (3.95±1.48)%,t=-3.865,P＜0.001].And the prevalence of left ventricular dyssynchrony was higher than that in control subjects(43% vs 3%).Conclusions HHC patients may have regional left ventricular systolic dysfunction before global changes,and have a higher prevalence of left ventricular dyssynchrony.RT-3DE is a useful imaging modality for assessing left ventricular systolic function.

Objective To explore and integrate the key techniques used in the surveillance and forecast of schistosomiasis in the water regions along the Yangtze River,so as to provide technical support for identifying rapidly the risk of schistosomiasis transmission and implementing control measures targeting the risk. Methods According to the distribution of water systems and water regions along the Yangtze River in Jiangsu Province,the demonstration sites for surveillance and forecast of schistoso? miasis were set across the province,where the integration and demonstration of the techniques regarding monitoring of Schistoso?ma japonicum infection in sentinel mice,human and animal activities,release of forecast information,and emergency treat?ment of water regions at risk of infection were performed. The pattern of human and animal activities was compared with the S. ja?ponicum infection in sentinel mice in the demonstration sites,and the operability of the release of information and emergency treatment of the risk of S. japonicum infection was evaluated. Results A total of 50 demonstration sites for surveillance and forecast of schistosomiasis were set in fixed anchor points,opening of the navigation lock to the Yangtze River,freight terminal, agritainment places,ferry,large construction places,and places for guaranteeing the Youth Olympic Games in 23 counties(dis?tricts)of 5 cities,Jiangsu Province. During the period between May and September,2014,the infectivity of water body was monitored by using 5 batches of sentinel mice,with a 99.06%(4 954/5 001)gross recovery rate of mice. S. japonicum infection was detected in a demonstration site,and an infected mouse was found,with a 0.02%(1/4 933)gross positive rate of sentinel mice. The field survey showed 2 088 person?times contacting water,including 91.95%(1 920/2 088)contacting water due to the production such as capturing fish,harvesting and cultivating crops,and repairing and building boats,and 8.05%(168/2 088)contacting water due to the life activity,such as fishing,washing vegetables and playing with water. The people contacted water predominantly in August and September(49.57%). A total of 859 boats containing 1 877 boatmen were observed,68.22%(586/859)of which were fishing boats containing 1 306 fishermen(69.58%). There were 32 sheep found in 4 demonstration sites,and 3 sheep were seen in the demonstration site with infected sentinel mouse. Four blue forecasts(emergence of water con?tact)and one orange forecast(S. japonicum?infected sentinel mouse detected)were released across the province,with one fore?cast map released which showed 5 sites with fishing and one site with sheep grazing,one emergency response system initiated, mollusciciding implemented in 10 hm2 high?risk regions,120 sheep grazed in fence,and 35 fishermen given health?education materials,schistosomiasis examination and preventive therapy. In addition,no acute schistosomiasis occurred in the demonstra?tion site with S. japonicum?infected sentinel mice. Conclusions The integration and demonstration of the techniques regarding monitoring of S. japonicum infection in sentinel mice,human and animal activities,release of forecast information,and emer?gency treatment of water regions at risk of infection provides an effective approach for the large?scale surveillance and forecast of schistosomiasis.

OBJECTIVETo explore the incidence and location of Y chromosome microdeletions in Chinese azoospermia and severe oligozoospermia, as well as the relationship between the deletion region and testicular phenotype.

METHODSSemen samples or blood samples were collected from 664 Chinese patients (584 with azoospermia and 80 with severe oligozoospermia). DNA was extracted by incubating cells with a lysis buffer containing polymerase chain reaction (PCR) buffer and proteinase K, and was assayed for deletion of 15 sequence tagged sites (including 6 loci recommended by European Academy of Andrology and European Molecular Genetics Quality Network (EAA/EMQN) distributed in AZFa, AZFb and AZFc by 4 multiplex PCRs. The histological phenotypes of testes of some azoospermic patients harboring Y chromosome microdeletion were studied by fine needle aspiration.

RESULTSSixty-six (11.3%) cases of microdeletions were found in the 584 patients with azoospermia, and deletions of AZFc region are the leading group (72.7% of all deletions), followed by AZFbc (13.6%), AZFabc (6.1%), AZFb (4.5%) and AZFa (3.0%). In the 80 men with severe oligozoospermia, 10 (12.5%) cases of AZFc microdeletions were detected. While azoospermia (n=19) with AZFc region deletion showed variable testicular phenotype, deletions of AZFb+c and AZFa+b+c (n=7) resulted in severe impaired spermatogenesis characterized by Sertoli cell only syndrome and spermatogenic arrest at spermatogonia.

CONCLUSIONIn the Chinese men with azoospermia and severe oligozoospermia, the incidence of Y chromosome microdeletions and the frequency of the deletions of the three AZF regions are similar to those described previously in other populations. Massive deletions of AZFb+c and AZFa+b+c impair spermatogenesis severely.

Azoospermia ; genetics ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Humans ; Male ; Models, Genetic ; Oligospermia ; genetics ; Polymerase Chain Reaction

OBJECTIVETo investigate the result of zygomatic reduction with midface L-shaped osteotomy through intraoral approach.

METHODSFrom June 2006 to Aug. 2009, 67 cases received zygomatic reduction with midface L-shaped osteotomy through intraoral approach. 52 cases underwent CT scan before operation and 12 months after operation. The images were analyzed by software GE AW 4.1 for evaluation of clinical effect, maxillary sinus change and complication. SAS 6.12 software was applied for one-way ANOVA.

RESULTSSatisfactory results were achieved in all the patients. The volume of maxillary sinus at left and right side was (21233.96 +/- 4455.04) mm3, and (22020.64 +/- 3663.82) mm3, respectively before operation: (17840.91 +/- 4381.03) mm3 and (18511.85 +/- 3466.24) mm3 respectively 12 months after operation, showing a significant difference between them (P<0.05). No infection or dental pulp necrosis happened.

CONCLUSIONSGood results can be achieved with intraoral L-shaped osteotomy for zygomatic reduction. Exposure of maxillary sinus would not cause any complication.

Adult ; Female ; Humans ; Male ; Maxillary Sinus ; Osteotomy, Le Fort ; adverse effects ; methods ; Postoperative Complications ; epidemiology ; Reconstructive Surgical Procedures ; methods ; Young Adult

OBJECTIVETo evaluate the clinical performance of an innovative contrast-enhanced ultrasound (CEUS) technique in characterization of focal liver lesions (FLL).

METHODSCEUS was performed in 505 patients with FLLs. A sulfur hexafluoride-filled microbubble ultrasound contrast agent and a continuous real-time imaging technique of contrast pulse sequencing were used. The observation items included initial nodule enhancement time, enhancement level and pattern, and dynamic evolution of enhancement along with the CEUS phases.

RESULTSThere were similar findings for both malignant and benign FLLs in initial enhancement time and enhancement level on CEUS. The dynamic evolution of enhancement along with enhancement phase was the most significant parameter for characterization of FLLs. Most malignant lesions showed enhancement at arterial phase and hypo-enhancing at late phase, whereas the benign lesions presented enhancement at arterial phase and persistent hyper- or iso-enhancing until late phase. Using these characteristics as a diagnostic criteria, the sensitivity, specificity, positive predicative value, negative predicative value and accuracy of CEUS in identifying malignant FLLs were 95.7% (313/327), 86.0% (153/178), 92.6% (313/338), 91.6% (153/167) and 92.3% (466/505), respectively.

CONCLUSIONSCharacterization of FLLs by using the innovative CEUS technique is recommendable due to its safety, facility and high accuracy.

Adult ; Aged ; Aged, 80 and over ; Diagnosis, Differential ; Female ; Humans ; Liver Diseases ; diagnostic imaging ; Male ; Middle Aged ; Sulfur Hexafluoride ; Ultrasonography ; methods