1.The effect of Smad4 for normal development of mouse lacrimal gland
Ying, LIU ; Xin-zu, GU ; Michael, LU ; Chuan-qing, DING
Chinese Journal of Experimental Ophthalmology 2012;30(5):397-402
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.
2.Structure and function of 3'- untranslated region in picornavirus.
Rui-Ying LIANG ; Chuan-Feng LI ; Chun-Chun MENG ; Zong-Yan CHEN ; Guang-Qing LIU
Chinese Journal of Virology 2014;30(4):463-469
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
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Animals
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Humans
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Nucleic Acid Conformation
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Picornaviridae
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chemistry
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genetics
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metabolism
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Picornaviridae Infections
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virology
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RNA, Viral
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chemistry
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genetics
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metabolism
3.Application of five formulas in the elderly cataract patients with long axial length
Wei, FANG ; Jian, ZHANG ; Da-Chuan, LIU ; Wei-Jia, DAI ; Hui-Qing, YANG
International Eye Science 2017;17(7):1249-1253
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(24.5mm
4.Gene cloning, soluble expression and activity analysis of rSEA.
Quan-Bin XU ; Chuan-Xuan LIU ; Qing-Jun MA
Chinese Journal of Biotechnology 2003;19(4):402-406
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
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Cell Proliferation
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drug effects
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Cells, Cultured
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Cloning, Molecular
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Electrophoresis, Polyacrylamide Gel
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Enterotoxins
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genetics
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metabolism
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pharmacology
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Humans
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Leukocytes, Mononuclear
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cytology
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drug effects
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Plasmids
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Polymerase Chain Reaction
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Solubility
5.Preliminary research on the pathological role of cathepsin-B in subcutaneous heteroplastic pancreatic carcinoma in nude mice.
Chong ZHANG ; Jia-bang SUN ; Da-chuan LIU ; Ye-qing CUI ; Shuang LIU ; Hai-chen SUN
Chinese Medical Journal 2009;122(20):2489-2496
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
6.Experiment on autologous free granular fat transplantation with rhVEGF gene in rats.
Ming LEI ; Shi-Qing LIU ; Yu-Lan LIU ; Jun REN ; Chuan-Xun YI
Chinese Journal of Plastic Surgery 2008;24(1):67-70
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
7.Abbreviated MRI Protocols for Detecting Breast Cancer in Women with Dense Breasts.
Shuang Qing CHEN ; Min HUANG ; Yu Ying SHEN ; Chen Lu LIU ; Chuan Xiao XU
Korean Journal of Radiology 2017;18(3):470-475
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*
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Breast*
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Diagnosis
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Ethics Committees
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Female
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Humans
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Magnetic Resonance Imaging*
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Mammography
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Mass Screening
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Retrospective Studies
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Sensitivity and Specificity
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Transcription Factor AP-1
8.Algorithm of brain pathology detection based on statistical atlas of attribute vectors.
Chuan-fu LI ; Huan-qing FENG ; Weil LIU ; Zhen HAN ; Tao SUN
Chinese Journal of Medical Instrumentation 2009;33(1):7-10
To detect lesions of brain CT automatically, a statistical atlas of attribute vectors (SAAV) was designed and created to describe the multiple features of medical images. By comparing the features of study image with those of SAAV, we successfully detected various kinds of brain lesion. It was demonstrated that the algorithm is effective in detecting various kinds of lesions found on brain CT images. Further studies are needed to make the algorithm more acceptable.
Algorithms
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Brain Diseases
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diagnostic imaging
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Humans
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Image Processing, Computer-Assisted
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Numerical Analysis, Computer-Assisted
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Tomography, X-Ray Computed
10.Resveratrol reduces intracellular free calcium concentration in rat ventricular myocytes.
Zheng LIU ; Li-Ping ZHANG ; Hui-Jie MA ; Chuan WANG ; Ming LI ; Qing-Shan WANG
Acta Physiologica Sinica 2005;57(5):599-604
Resveratrol (trans-3, 4', 5-trihydroxy stilbene), a phytoalexin found in grape skins and red wine, has been reported to have a wide range of biological and pharmacological properties. It has been speculated that resveratrol may have cardioprotective activity. The objective of our study was to investigate the effects of resveratrol on intracellular calcium concentration ([Ca(2+)](i)) in rat ventricular myocytes. [Ca(2+)](i) was detected by laser scanning confocal microscopy. The results showed that resveratrol (15~60 mumol/L) reduced [Ca(2+)](i) in normal and Ca(2+)-free Tyrode's solution in a concentration-dependent manner. The effects of resveratrol on [Ca(2+)](i) in normal Tyrode's solution was partially inhibited by pretreatment with sodium orthovanadate (Na3VO4, 1.0 mmol/L, P<0.01), an inhibitor of protein tyrosine phosphatase, or L-type Ca(2+) channel agonist Bay K8644 (10 mumol/L, P<0.05), but could not be antagonized by NO synthase inhibitor L-NAME (1.0 mmol/L). Resveratrol also markedly inhibited the ryanodine-induced [Ca(2+)](i) increase in Ca(2+)-free Tyrode's solution (P<0.01). When Ca(2+) waves were produced by increasing extracellular Ca(2+) concentration from 1 to 10 mmol/L, resveratrol (60 mumol/L) could reduce the velocity and duration of propagating waves, and block the propagating waves of elevated [Ca(2+)](i). These results suggest that resveratrol may reduce the [Ca(2+)](i) in isolated rat ventricular myocytes. The inhibition of voltage-dependent Ca(2+) channel and tyrosine kinase, and alleviation of Ca(2+) release from sarcoplasmic reticulum (SR) are possibly involved in the effects of resveratrol on rat ventricular myocytes. These findings could help explain the protective activity of resveratrol against cardiovascular disease.
Animals
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Calcium
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metabolism
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Calcium Channels
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drug effects
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Heart Ventricles
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cytology
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metabolism
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Intracellular Fluid
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drug effects
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metabolism
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Male
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Myocytes, Cardiac
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drug effects
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metabolism
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Protein-Tyrosine Kinases
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drug effects
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Rats
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Rats, Sprague-Dawley
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Sarcoplasmic Reticulum
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drug effects
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metabolism
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Stilbenes
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pharmacology