1.Expression of recombinant plasmid of homo heterogeneous nuclear ribonucleoprotein E1 in SH-SY5Y cells
Lirong HUO ; Lanying WANG ; Junhua ZOU ; Nan ZHONG
Chinese Journal of Tissue Engineering Research 2014;(42):6769-6774
BACKGROUND:The functions of homo heterogeneous ribonucleoprotein E1 are very wide. It can participate in the expression of skeleton proteins in the nervous system. OBJECTIVE:To construct the recombinant plasmid of homo heterogeneous ribonucleoprotein E1 and observe its expression in nerve cells for further studying the functions of it in neurocytes. METHODS:Using pcDNATM4/His C, the homo heterogeneous ribonucleoprotein E1 was subcloned into recombinant plasmid E1-pcDNATM 4/His C, fol owed by enzyming and sequencing. After SH-SY5Y cells were transfected with the recombinant plasmid, western blot analysis and real time RT-PCR were used to detect the expression of homo heterogeneous ribonucleoprotein E1 in SH-SY5Y cells. And the growth of SH-SY5Y cells was observed. RESULTS AND CONCLUSION:We successful y constructed the eukaryotic expressed vector of homo heterogeneous ribonucleoprotein E1. The recombinant plasmids were verified to express in SH-SY5Y cells correctly at mRNA and protein levels. And SH-SY5Y cells generated quickly after homo heterogeneous ribonucleoprotein E1 was over-expressed. The homo heterogeneous ribonucleoprotein E1 is an important protein in neural development. And this vector offers the premise for further studying its function in nervous system.
2.MRI and pathological features of different molecular subtypes of breast cancers
Yang YU ; Tianlong HUO ; Yunyao LAI ; Nan HONG
Chinese Journal of Radiology 2014;48(3):184-188
Objective To investigate the MRI and pathological features of different molecular subtypes of breast cancer.Methods The data of 202 patients who underwent primary breast cancer resection were retrospectively reviewed.All of the patients had MRI preoperatively.The molecular subtypes of breast cancer defined by immunohistochemistry were classified as basal-like,luminal and HER-2 overexpression.Morphology (including mass or non-mass like enhancement,shape and margin of masses,unifocal or multifocal masses) and enhancement characteristics on MRI,histologic types and grades of tumors were analyzed with Chi-square test,exact test,Fisher exact test,Kruskal-Wallis H test,and Wilcoxon test.Results Among the 202 patients,34 were basal-like,144 were luminal and 24 were HER-2 overexpression.The number of mass cases in each subtype was 29,133 and 19 respectively,making no significant difference (x2 =4.136,P =0.126).As for the shape of basal-like lesions,8 were round,19 were lobular and 2 were irregular,while this distribution was 23,58,52 in luminal subtype and 1,11,7 in HER-2 overexpression subtype (x2 =13.391,P < 0.05).The margin was also strikingly different among three groups (smooth,spiculated,irregular):20,5,4 respectively in basal-like,27,53,53 respectively in luminal,and 4,7,8 respectively in HER-2 overexpression (x2 =28.515,P < 0.01).52.6% (10/19) of HER-2 overexpression cases were multifocal,while only 6.9% (2/29) of luminal and 8.0% (24/133) of basal-like ones were multifocal (x2 =16.140,P < 0.01).Characteristics in dynamic contrast-enhanced MRI were statistically different,with homogeneous,heterogeneous,and rim enhancement 0,13,16 respectively in basal-like cases,28,93,11 respectively in luminal cases and 2,11,6 respectively in HER-2 overexpression cases (P < 0.01).However,the difference for enhancement curve did not reach significance (P =0.457).Histologic types were significantly different among molecular subtypes (P < 0.01).Luminal breast cancer consisted of various histologic types,32.6% (47/144) of which were mixed type.The majority of the other two molecular subtypes were invasive ductal carcinoma.Furthermore,invasive ductal carcinomas with different molecular subtypes showed different histologic grades (Hc =30.014,P < 0.01).Basal-like breast cancer was more likely associated with a higher grade of malignancy.Conclusions Different molecular subtypes of breast showed distinct MRI features and pathologic characteristics.MRI might be a useful tool for preoperative prediction of molecular subtypes of breast cancer.
3.The Application Practice of Movie Appreciation Curriculum in HIV/AIDS Education for College Students
Qingqing HE ; Lu CHEN ; Mengxiao WEI ; Nan HUO ; Feng JIANG
Chinese Medical Ethics 1996;0(01):-
In current years,there is a tendency for HIV/AIDS to shift to adolescent population,thus HIV/AIDS education aimed at college students is drawing increasing attention from all parts including the governments,colleges and universities,and the whole society.By analyzing the current spreading trend of HIV/AIDS,this thesis introduces the modern teaching methods in HIV/AIDS education and existing problems,and tentatively introduces movie appreciation as a novel teaching approach.The application advantages of movie appreciation and concrete examples from six universities in Shanxi Provinces have also be delivered,aiming to explore a new pattern for HIV/AIDS education among Chinese college students.
4.Protective effect of shenfu injection against ischemia-reperfusion injury due to pancreas transplantation in rats.
Xiao-nan LIU ; Ting-ting HUO ; Wei-zhong WANG
Chinese Journal of Integrated Traditional and Western Medicine 2006;26 Suppl():111-115
OBJECTIVETo investigate the protective effect of Shenfu Injection against ischemia-reperfusion (I/R) injury due to pancreas transplantation in rats, and explore its possible mechanism.
METHODSSix normal SD rats with sham operation were taken as the normal control group, 24 steptozozin-induced diabetic SD rats were randomly divided into 4 groups, with 6 in each group. Except I/R group, the rats in the other groups were intravenous injected with Shenfu Injection (SF,10 mg/kg), Hongshen Injection (HS, 9 mg/kg) and Fuzi Injection (FZ 1 mg/kg) respectively at the day and 30 minutes before pancreas transplantation performed in the SF group, HS group and FZ group, respectively. At the same time, rats in the normal control group and in the I/R group were intravenously injected the same volume of normal saline. The blood glucose was detected before and after reperfusion, and 2 hours later after reperfusion, the contents of serum nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha), the concentrations of malondialdehyde (MDA) , superoxide dismutase (SOD) , and myeloperoxidase (MPO) in the transplanted pancreas tissues were detected. The cell apoptosis of the transplanted pancreas tissue was determined by TUNEL, and the bcl-2 and Bax protein expression was determined by Western blot.
RESULTSAfter reperfusion, the levels of blood glucose and TNF-alpha decreased and the concentration of NO increased in the SF group, HS group and FZ group, compared with those in the I/R group. The activity of SOD, bcl-2 expression and the ratio of bcl-2 and Bax were higher, while the content of MDA, the activity of MPO, apoptotic indexes, and Bax expression were lower in the SF group, HS group and FZ group than those in the I/R group.
CONCLUSIONShenfu Injection can protect L/R injury due to pancreas transplantation in rats, the possible mechanism may be related to promoting activity of SOD, increasing synthesis of endogenous NO, decreasing the excretion of TNF-alpha, alleviating conglutination and aggregation of polymorphonuclear neutrophils (PMNs) in pancreas, as well as up-regulating Bcl-2 gene expression and down-regulating the Bax gene expression.
Animals ; Cell Aggregation ; drug effects ; Diabetes Mellitus, Experimental ; enzymology ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Injections ; Malondialdehyde ; metabolism ; Nitric Oxide ; blood ; Pancreas ; drug effects ; metabolism ; Pancreas Transplantation ; adverse effects ; Protective Agents ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; prevention & control ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism
5.Rapid screening of katG gene mutation in isoniazid-resistant Mycobacterium tuberculosis
Journal of Zhejiang University. Medical sciences 2002;31(3):178-180
OBJECTIVE: To evaluate the relationship between katG gene mutation and isoniazid (INH) resistance and to develop a rapid screening method of point mutation in the katG gene associated with MTB resistance. METHODS: Twenty-four clinical isolates of MTB with 8 INH resigtance isolates and 16 INH-sensitive isolates were analyzed by PCR-RFLP, with the H(37)Rv reference strain as the control. RESULTS: G-->C point mutations were detected in 7 of 8 isoniazid-resistant strains and no gene mutation was shown in 16 isoniazid-sensitive isolates. The sensitivity and specificity were 87.5 % and 100 % respectively. No katG gene sequence deletion was observed in any specimen. CONCLUSION: Our results suggest katG gene mutation is one of the most important mechanisms of INH-resistant TB. PCR-RFLP may be useful in detection of katG gene mutation.
6.The inhibitory effect of decorin on proliferation of rabbit epithelial cell
Jian-nan, XIANG ; Gui-lan, ZHANG ; Hai-jiang, ZHANG ; Guo-hua, WANG ; Ming, HUO
Chinese Journal of Experimental Ophthalmology 2012;30(1):41-45
BackgroundResearches found that the posterior capsular opacification (PCO) after lensextraction is associated with the elevation of the transforming growth factor-β(TGF-β).To seek the drug for inhibitingproliferation of lens epithelial cells (LECs) is crucial in the treatment and prevention of PCO.ObjectiveThisstudy was to investigate the preventing effects of decorin on the proliferation of LECs.MethodsRabbit LECs wascultured and passaged.The LECs in growth phase were incubated in 96 well plate at the density of 8×106/L.Decorinwith the concentrations 0.1,1.0,10.0 mg/L was added into the medium for 24,48 and 72 hours respectively.0.1%DMSO was used at the same way as positive control group,and the regular cultured cells worked as blank controlgroup.The inhibitory rates of different concentrations of decorin on the growth of LECs were detected by MTT at 24,48and 72 hours after addition of decorin.The percentage of LECs in different cell cycles in various groups was assayedusing flow cytometry.TGF-β level in medium suspension was detected using ELISA.The expression of TGF-β mRNA in LECs was checked by RT-PCR,and α-SMA expression in LECs was determined using immunochemistry.Results ELISA assay showed a statistical difference in the TGF-β levels of different groups (F=39.24,P=0.03 ).The TGF-β levels in 1.0,10.0 mg/L decorin groups were significantly decreased in comparison with blank control group (P<0.01) and 0.1 mg/L decorin group (P<0.05 ).The inhibitory rates of decorin in the concentrations of ≥ 1.0 mg/L on the growth of LECs were higher than the blank control group,and those in various concentrations of decorin groups were considerably lower in 24 hours compared with 48 and 72 hours ( P<0.05 ) and so was the 48 hours compared with 72 hours (P<0.05 ).The percentages of LECs in G0/G1 phase were ascent in 0.1,1.0 and 10.0 mg/L decorin groups in comparison with G2/M and S phase (P<0.05).Immunochemistry revealed the weak expression of α-SMA in various decorin groups in comparison with control group. Conclusions Decorin can effectively inhibit LECs growth and induce LECs apoptosis in concentration- and time-dependent manner.It is suggested that decorin can be used in the prevention and treatment of after cataract.
7.Effects of iron overload on cognitive function and autophagy of hippocampal neurons in rats
Tiantian HUO ; Jingru ZHAO ; Nan MENG ; Lipeng DONG ; Litao LI ; Peiyuan LYU
Chinese Journal of Behavioral Medicine and Brain Science 2021;30(1):9-14
Objective:To explore the effect of iron overload on the cognitive function of rats and its possible internal mechanism.Methods:Thirty 8-week-old male Sprague Dawley rats of SPF degree were randomly divided into 2 groups, iron overload group(IO group) and control group(Sham group), with 15 in each group.The rats in IO group were injected intraperitoneally iron dextran(100 mg/(kg·d)) for 28 days.The cognitive function of rats was detected by Morris water maze method. Western blot method was used to detect the expression of TfR1 and autophagy-related protein p-AMPK, LC3 and Beclin1 in the hippocampus of rats. Immunofluorescence was used to observe the expression of LC3 and Beclin1 in the hippocampus of rats. HE staining was used to observe the morphological changes of neurons in the hippocampus. Transmission electron microscopy was used to observe the number of autophagosomes and the morphology of endoplasmic reticulum in hippocampus.The software of SPSS 20.0 was used for repeated measurement ANOVA and t-test. Results:Morris water maze test showed that there were significant interaction between the group factor and training time factor of escape latency( F=3.55, P<0.01). And the simple effect analysis showed that compared with the Sham group((28.09±18.41)s, (21.42±15.53)s, (16.96±8.35)s, (10.24±3.75)s), the average escape latency of rats(2nd-5th day) in IO group((56.68±30.65)s, (58.21±36.09)s, (36.58±13.54)s, (27.29±14.30)s )were significantly longer ( t=8.57, 6.81, 9.51, 7.12, P<0.01). The platform was removed on 6th day of the space exploration experiment, compared with the Sham group ((41.89±3.89)%), the percentage of time spent in the target quadrant of IO group ((25.46±3.56)%) was significantly decreased( t=24.06, P<0.01). Western blot showed that the relative expression levels of (TfR1 (2.10±0.48), p-AMPK (0.74±0.10), LC3 (1.11±0.40), Beclin1 (1.05±0.20)) in IO group in the hippocampus of the rats were significantly higher than those of the Sham group(TfR1(0.11±0.18), p-AMPK(0.19±0.02), LC3(0.22±0.11), Beclin1(0.17±0.02))( t=1.58, 14.58, 10.06, 20.65, P<0.01)). HE staining showed that compared with the Sham group, the neuron in the hippocampus of the IO group were sparsely arranged, morphologically irregular, and the number of the neurons was significantly reduced. Transmission electron microscopy showed that compared with the Sham group, the number of autophagosomes in the hippocampus of IO group was increased. Conclusion:Iron overload may exert its neurotoxic effect by increasing the level of autophagy in the hippocampus, causing cognitive dysfunction.
8.Clinical experience of penile augmentation with superficial fascia retrocession: a report of 60 cases.
Xia LI ; Hai-Tao WANG ; Jin-Cun YANG ; Ran HUO ; Cheng ZHANG ; Nan HU
Chinese Journal of Plastic Surgery 2013;29(1):34-37
OBJECTIVETo investigate the clinical effect of penile augmentation with superficial fascia retrocession.
METHODSThe first annular incision line was designed, reserving the inner plate 0.5 -0.8 cm. The second annular incision line was determined, according to the length of penile erection. The skin tissue between the double loops was stripped, reserving the superficial fascia and the superficial dorsal veins. Before the closure of two annual incision with no tension, the superficial fascia tissue was pushed proximally and fixed sectionally to augment penis.
RESULTSFrom Jun 2009 to Jun 2011, 60 patients with redundant prepuce or phimosis were treated with this method. During 3 to 12 months follow-up, the postoperative penile perimeter increased significantly. The glans were exposed and the prepuce looked smooth. The function of the penis maintained undamaged.
CONCLUSIONSThe penile augmentation with superficial fascia retrocession can make full use of autologous tissue to augment penis, meanwhile, redundant prepuce can be resected. It is very practical for clinical application.
Adolescent ; Adult ; Circumcision, Male ; methods ; Humans ; Male ; Middle Aged ; Penis ; surgery ; Subcutaneous Tissue ; surgery ; Transplantation, Autologous ; Young Adult
9.Effect of pigment epithelium derived factor on NO and the expression of caspase-3 in retinal tissues of model rats with optic nerve crush injury
Xiao-Xiao, YAN ; Hai-Bo, JIA ; Xiao-Ling, YIN ; Cui, CUI ; Wei-Xing, PU ; Nan, HUO ; Jun-Bo, ZHAO
International Eye Science 2017;17(6):1047-1050
AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.
10.Advances in research on therapeutic strategies of targeting pancreatic cancer stroma
Xinyuan ZHOU ; Nan LIU ; Pan ZHANG ; Meirong HUO
Journal of China Pharmaceutical University 2021;52(6):653-662
Pancreatic cancer stroma plays a critical role in tumor progression, invasion, metastasis and resistance.Targeting tumor cell alone could not meet the demand for prolonging patients'' survival.Growing studies have laid emphasis on developing combined regimens between targeting pancreatic cancer stroma and chemotherapy, radiotherapy and immunotherapy.We are faced with some new opportunities in spite of the great challenges brought to the research and development of targeting drugs owing to the complicated stroma components, crosstalking signal pathways and abnormal angiogenesis of pancreatic cancer.In this article, recent advances in therapeutic strategies of targeting pancreatic cancer stroma are reviewed and analyzed from the aspects of extracellular matrix (ECM), cancer associated fibroblasts (CAFs) and vessels, in the hope of providing some novel ideas for targeting therapy against pancreatic cancer.