BackgroundSmad4,a key intracellular mediator in transforming growth factor-β (TGF-β)signaling,plays a critical role in the normal development of many tissues/organs.However,its functional role in the development of lacrimal gland has rarely been reported.ObjectiveThe aim of this study was to investigate the role that Smad4 may play in the development of lacrimal glands using Smad4 conditional knockout (CKO) mice( C57BL/6 mouse line),MethodsSmad4 in lacrimal glands,as well as in the lens,cornea and ectoderm of the eyelids,was conditionally inactivated by the Pax6 promoter-driven Cre transgenic mice and Smad4 conditional gene mice,LacZ reporter was used to visualize the developing lacrimal gland by X-gal staining,and standard histological approaches were used to reveal morphological changes.Six or more mice or embryos in each group were used for comparisons at the same stage.ResultsLacZ staining showed that E15.0,Smad4 CKO mice could still develop primary lacrimal bud,but much shorter than the wild-type ones.At E16.5,the primary lacrimal bud in wild-type mice began branching,but no branching was found in Smad4 CKO mice except that the primary lacimal bud became blunt at the tip.At E18.0,although Smad4 CKO mice develop some acini,the branching and size and number of acini were obviously less than ones in Smad4 wild-type mice.Based on histological findings,lacrimal glands in Smad4 CKO mice developed slowly,and the size was considerably smaller,and the numbers of lobes as well as the numbers of acini were much fewer than those of Smad4 wild-type mice lacrimal glands at various stages.Pigment and adipose tissue were also observed within the lacrimal glands starting from P7 in Smad4 CKO mice and increased with age growing.Lacrimal glands in mutant adult mice were eventually replaced by adipose tissue and accumulation of pigments.Conclusions These results support the notion that Smad4 is essential for the normal development and maintenance of the mouse LG and may be involved in the metabolism of pigment and adipose tissue in LG.

Both sides of the picornavirus genome have 5'-untranslated region (5'UTR) and 3'- untranslated region (3'UTR). This study demontrated that both the 5'-and 3'-UTR can form complex structures, such as stem-loop, clover and pseudoknot structure, These structures play an important role in the regulaton of the replication and translation of the viruses. This article reviewed the progress of research on the structure and function of picornavirus' 3'-UTR over recent years.
3' Untranslated Regions ; Animals ; Humans ; Nucleic Acid Conformation ; Picornaviridae ; chemistry ; genetics ; metabolism ; Picornaviridae Infections ; virology ; RNA, Viral ; chemistry ; genetics ; metabolism

AIM: To compare the accuracy of intraocular lens(IOL)power calculations by using five formulas(Haigis, SRK-T, Hoffer Q, Holladay-1, SRK-Ⅱ)in eyes with long axial lengths in order to improve the accuracy of predicating IOL powers.METHODS: Fifty-one eyes of 51 cases of age-related cataract and with mild long axial(24.5mm27mm) were collected who`s optical biometry were performed by the Zeiss IOL Master500 before operation.They underwent regular phacoemulsification and posterior chamber IOL implantation.The actual postoperative refraction was measured with the methods of phoropter and subjective optometry 3mo after surgery.Then we compared the differences of the predicted and actual postoperative refraction of the five formulas in each group.RESULTS: In the mild axial lengths cases, the differences between SRK Ⅱ formula and the other four formulas were statistically significant (P<0.05), and the difference between Hoffer Q and SRK-T formula was statistically significant (P<0.05);there was no difference among the other formulas (P>0.05).In the moderate and severe long axial lengths cases, the differences between SRK Ⅱ formula and the other four formulas were statistically significant (P<0.05), and the difference between Hoffer Q and SRK-T formula, Hoffer Q and Haigis formula were statistically significant (P<0.05);there was no difference among the other formulas (P>0.05).The differences of all the five formulas between the two groups were statistically significant (P<0.05).CONCLUSION: In the mild axial lengths cases, Haigis, SRK-T, Hoffer Q, Holladay-1 performed well.In the moderate and severe long axial lengths cases, Haigis, SRK-T and Holladay-1 performed better than other formulas.The accuracy of all the five formulas decreases as the axial length getting longer.

The superantigen,such as staphylococcal enterotoxins, had been identified as possible anti-cancer molecules in many reports. In this paper, we cloned the entA gene encoding Staphylococcal enterotoxin A from the genomic DNA of Staphylococcus aureus(ATCC13565) by PCR, the sequence cloned was accordance with that reported in Genebank. The entA gene could be expressed effectively after inserted into plasmid pET-22b( + ), The rSEA was expressed as inclusion bodies when induced by IPTG at 37 degrees C and became soluble after induced at low temperature, the soluble part is about 55% of total rSEA products. Only one band was detected by western-blotting in expression product of BL-21 (DE3) with pET-SEA. The soluble rSEA was purified by Ni2+ chelating sepharose column. No other protein except rSEA was seen in SDS-PAGE gel stained by both Coomassie brilliant blue and silver salt, which showed that the rSEA was purified effectively. Homology modeling of rSEA determined the structure change was conducted, which indicated there was no apparent structure change between rSEA and native SEA. This result was also confirmed by proliferation assay of PBMC, for the rSEA could induced proliferation of PBMC as effectively as native SEA. The increasing anti-tumor activity of rSEA was also detected after the spleen cell activated in vivo by rSEA, which was accordance with others reports. This work paved the way for the further study of anti-cancer with rSEA.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Enterotoxins ; genetics ; metabolism ; pharmacology ; Humans ; Leukocytes, Mononuclear ; cytology ; drug effects ; Plasmids ; Polymerase Chain Reaction ; Solubility

BACKGROUNDCathespin-B (cath-B) is an important proteolytic enzyme involved in the disease course of invasion in many types of cancer. Cath-B expression in subcutaneous heteroplastic pancreatic carcinoma in nude mice has not been studied. We investigated the role of cath-B in a model of heteroplastic pancreatic carcinoma in BALB/c nude mice.

METHODSThirty-two six-week-old female BALB/c nude mice were equally divided into four groups. PANC-1 cells were inoculated subcutaneously in the left axillary region. Besides volume, weight of subcutaneous tumor, and change in body weight, cath-B expression in each group was measured by immunohistochemical staining, PCR and Western blotting. Its relationship to microvessel density (MVD), CD44v6, and placenta growth factor (PLGF) was also examined. CA-074Me, a specific inhibitor of cath-B, was injected intraperitoneally (i.p.) at different stages of tumor growth in group B and C. Gemcitabine (GEM), was also injected (i.p.) in group D to compare anti-tumor efficacy with CA-074Me.

RESULTSExpression of cath-B at different levels was related to tumor growth, MVD, and PLGF expression. In group A (control group), cath-B expression was enhanced more than that seen in other groups. CA-074Me clearly inhibited cath-B expression and tumor growth in group B. There was no difference between group C and D with respect to anti-tumor effect.

CONCLUSIONSCath-B correlates with the growth and angiogenesis of tumors, but not with the adhesion induced by CD44v6. CA-074Me clearly inhibited cath-B expression and demonstrated an anti-neoplastic and anti-angiogenesis effect.

Animals ; Antineoplastic Agents ; therapeutic use ; Blotting, Western ; Body Weight ; Cathepsin B ; antagonists & inhibitors ; genetics ; metabolism ; physiology ; Cell Line, Tumor ; Dipeptides ; therapeutic use ; Female ; Humans ; In Vitro Techniques ; Mice ; Mice, Nude ; Pancreatic Neoplasms ; drug therapy ; metabolism ; Placenta Growth Factor ; Pregnancy Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Heterologous

OBJECTIVETo investigate the effects of rhVEGF on autologous free granular fat grafts in rats.

METHODSForty-eight Sprague-Dawley rats were randomly divided into three groups, sixteen of each. After the autologous free granular fat transplantation, all groups were treated with the plasmid DNA containing cDNA encoding rhVEGF, the blank plasmid DNA and normal saline respectively as the experimental group, the negative group and the saline group. After 3, 7, 15, 30 days, the rats were sacrificed and the grafts were weighted accurately. Histological pathology was evaluated. Micro-vessel count and the expression of vascular endothelial growth factor (VEGF) were examined by immunohistochemical staining.

RESULTSThe weights of the two latter groups were significantly reduced on the 7, 15, 30 day compared with the experimental group. The expression of VEGF and the micro-vessel count in the experimental group were significantly higher than the other two groups during the latter periods.

CONCLUSIONThe cDNA encoding VEGF can induce the expression of VEGF in fat graft, angiogenesis and reduce the free fat graft absorption.

Adipose Tissue ; transplantation ; Animals ; Female ; Gene Transfer Techniques ; Graft Survival ; Male ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; genetics ; Transplantation, Autologous ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVE: To evaluate the validity of two abbreviated protocols (AP) of MRI in breast cancer screening of dense breast tissue. MATERIALS AND METHODS: This was a retrospective study in 356 participants with dense breast tissue and negative mammography results. The study was approved by the Nanjing Medical University Ethics Committee. Patients were imaged with a full diagnostic protocol (FDP) of MRI. Two APs (AP-1 consisting of the first post-contrast subtracted [FAST] and maximum-intensity projection [MIP] images, and AP-2 consisting of AP-1 combined with diffusion-weighted imaging [DWI]) and FDP images were analyzed separately, and the sensitivities and specificities of breast cancer detection were calculated. RESULTS: Of the 356 women, 67 lesions were detected in 67 women (18.8%) by standard MR protocol, and histological examination revealed 14 malignant lesions and 53 benign lesions. The average interpretation time of AP-1 and AP-2 were 37 seconds and 54 seconds, respectively, while the average interpretation time of the FDP was 3 minutes and 25 seconds. The sensitivities of the AP-1, AP-2, and FDP were 92.9, 100, and 100%, respectively, and the specificities of the three MR protocols were 86.5, 95.0, and 96.8%, respectively. There was no significant difference among the three MR protocols in the diagnosis of breast cancer (p > 0.05). However, the specificity of AP-1 was significantly lower than that of AP-2 (p = 0.031) and FDP (p = 0.035), while there was no difference between AP-2 and FDP (p > 0.05). CONCLUSION: The AP may be efficient in the breast cancer screening of dense breast tissue. FAST and MIP images combined with DWI of MRI are helpful to improve the specificity of breast cancer detection.
Breast Neoplasms* ; Breast* ; Diagnosis ; Ethics Committees ; Female ; Humans ; Magnetic Resonance Imaging* ; Mammography ; Mass Screening ; Retrospective Studies ; Sensitivity and Specificity ; Transcription Factor AP-1

In this study, daphnetin and its major metabolites in the intestinal wall of rats were identified by liquid chromatography and quatrupole-time of flight mass spectrometry. Perfusion fluid of duodenum, jejunum, ileum and colon were collected separately for 2 hours from the rat intestine following perfusion with daphnetin. The metabolites of daphnetin in the perfusion fluid of different intestine segments were analyzed by the liquid chromatography and quatrupole-time of flight mass spectrometry. It is shown that the parent drug daphnetin and four metabolites were found in the perfusion fluid of duodenum, jejunum and ileum. However, no metabolites were found in the colon. Among the four metabolites, two daphnetin sulfates (m/z 257) were first discovered as the phase II metabolites of daphnetin in rats, which revealed a new way of daphnetin metabolism in rats.
Animals ; Chromatography, High Pressure Liquid ; Colon ; metabolism ; Duodenum ; metabolism ; Ileum ; metabolism ; Intestines ; metabolism ; Jejunum ; metabolism ; Male ; Perfusion ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization ; Umbelliferones ; metabolism ; pharmacokinetics

In early 2011, the serious outbreak of porcine pseudorabies virus (PRV) infection suddenly recurred in Henan and neighboring Provinces. To investigate the etiology of massive infection with PRV, 16 800 serum samples, 905 porcine epidemic diarrhea virus (PEDV) back-feeding tissues, and 56 PR gene deleted live vaccines were colleted from January 2011 to May 2013 to detect PRV field infection using a PRV gE antibody test kit. The gE and TK genes of 11 new epidemic PRV strains were sequenced by PCR, and their molecular characteristics were analyzed. Moreover, virus titer determination, protective test against PRV, and vaccine potency testing were performed. The results showed that the detection rate of PRV field infection-positive pig farms was 68.06%, and the overall positive rate of PRV field infection in serum was 38.47%; the positive rates in breeding sows, breeding boars, reserve pigs, and commercial pigs were 40.12%, 30.88%, 54.67%, and 26.52%, respectively. The new epidemic strains were in the same evolutionary branch and belonged to the virulent strain group. Compared with the classical PRV strain, the virulence of new epidemic strains changed a little. The length of gE gene was 1 787 bp, and the length of TK gene was 963 bp. The nucleotide homologies of gE and TK genes to Chinese reference strains were 98.2%-99.8% and 98.90%-99.6%, respectively, and the amino acid homologies were 97.1%-99.8% and 97.5%-99.4%, respectively. Commercial vaccine had a 100% protective effect against the new epidemic strains. The positive rate of PRV field infection was 0% in vaccine and 40.44% in back-feeding tissues. The results confirmed that PRV field infection rates were rising sharply among pigs in Henan and neighboring Provinces after 2011. The main virulence genes of new epidemic PRV strains did not change significantly over the years. PR gene deleted live vaccines had no PRV field infection and could completely resist the attack of new strains. The virus carriage of breeding boars and reserve pigs and the serious PRV field infection in PEDV back-feeding tissues were the main causative factors for massive infection with PRV and epidemic outbreak in Henan and neighboring Provinces from 2011 to 2013.
Amino Acid Sequence ; Animal Feed ; analysis ; virology ; Animals ; China ; epidemiology ; Epidemics ; Female ; Herpesvirus 1, Suid ; chemistry ; classification ; genetics ; isolation & purification ; Male ; Molecular Sequence Data ; Phylogeny ; Pseudorabies ; epidemiology ; virology ; Sequence Alignment ; Sequence Homology, Amino Acid ; Sus scrofa ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Proteins ; chemistry ; genetics

Encephalomyocarditis virus (EMCV) is a natural epidemic zoonotic pathogen. However, no reports have been published regarding the isolation, identification and full-length genome of EMCV from a local aardvark population. In present study, an EMCV isolate HNXX13 was isolated from aardvarks named Huainan-pig in Henan Province. The systematic identification, full-length genome sequencing and molecular characteristic analysis of the isolate HNXX13 were conducted. The result showed that the isolate was spherical with a diameter of 24-30 nm, neither heat- nor acid-resistant, sensitive to trypsin, insensitive to chloroform, not protected by bivalent cationic, and the specific fluorescence was observed in the cytoplasm of BHK-21 cells infected with the isolate by using indirect fluorescence assay. The full-length genome of EMCV HNXX13 generated a 7 725bp sequence (GenBank: F771002), with 81.0%-99.9% nucleotide identity to reference strains from different animals, and 99.5% with a Chinese reference strain isolated earlier from a commercial pig herd. The phylogenetic tree based on the full-length genome and ORF sequences identified that all EMCV strains were divided into three groups G1, G2 and G3, and strain HNXX13 belonging to the G1 group with other Chinese reference strains. The result also identified that this EMCV infection could cause severe clinical signs in a local aardvark population, and enriches the molecular epidemiological data of EMCV in China. Regional differences exist in EMCV genome and transmission is limited within a certain area. However, the cross-infection and transmission of EMCV between aardvark and mice appears most likely. Mutations have occurred in some amino acids of EMCV strain HNXX13 during the transmission in local aardvark herd and these mutations might make the virus easier to infect the aardvark.
Animals ; Animals, Wild ; virology ; Cardiovirus Infections ; veterinary ; virology ; China ; Encephalomyocarditis virus ; classification ; genetics ; isolation & purification ; Genome, Viral ; Mice ; Molecular Sequence Data ; Phylogeny ; Xenarthra ; virology