1.Specific TCRαβgene modification promotes anti-tumor reactivity of T cells
Fenglin WU ; Wenfeng ZHANG ; Mian HE ; Nuan YANG ; Han SHEN ; Huaben BO ; Hongwei SHAO ; Shulin HUANG
Chinese Journal of Immunology 2014;(7):901-908
Objective:To investigate effects of tumor specific TCR gene Vα12.2-Vβ7.1 modification on recognition of tumor antigen and activation of anti-tumor reactivity of T cells.Methods: T cells were transduced using recombinant Ad 5F35-TRAV-TRBV adenovirus ,and multiplicity of infection was optimized.Specific lysis of T cells was evaluated by calcein release assay.The frequency of apoptotic cells in target cells was detected by Annexin V /PI double-labeled FACS.The expression of FasL on T cells was analyzed by FACS.The secretion of cytokine IFN-γand IL-2 of T cells was determined by ELISA assays.Results: The highest tranduce efficiency was obtained at MOI 100 by recombinant Ad5F35-TRAV-TRBV adenovirus.The frequency of TCRVα12+Vβ7+cells reached above 25%3 days after transduction.TCR gene modification enhanced the ability of T cells to lyse HLA-A2+AFP+target cells(P<0.001), the ability of T cells to induce HepG-2 apoptosis(P<0.001),and expression of FasL on T cells(P<0.001).TCR gene modification also enhanced T cells to secret IFN-γafter coculture with antigen positive tumor cells ( P<0.001 ).Conclusion: Specific TCR gene modification by recombinant adeno virus effectively promotes T cells to recognize antigen positive tumor cell and exert anti -tumor reac-tivity.
2. Advances in study on regulation of hair pigment expression by natural products
Chinese Traditional and Herbal Drugs 2015;46(9):1393-1398
Hair color is affected not only by genetic, age, and environmental factors, but also by the regulation of many cytokines and proteins. The generation of melanin is mainly regulated by POMC, α-MSH, MC1R, MITF, tyrosinase, TRP-1, TRP-2, ASIP, etc. Hair color anomaly is often considered to be a feature of aging which brings great psychological pressure to the people, but the treartment by relevant drugs is still blank. This article summarizes the effect target and the function ways of those natural medicines with great efficacy on the treatment of hair graying. This review may provide some theoretical basis for the application of natural medicines in the treatment of hair graying.
4.Effect and prognostic factors of OIF and TIP in children with hypospadias
Wenbo HE ; Chuanbing HU ; Junyan TIAN ; Jinsong SUN ; Nuan HAN
Journal of Clinical Surgery 2023;31(11):1094-1098
Objective To explore the efficacy and prognostic independent factors of modified island flap(OIF)and urethroplasty(tip)in the treatment of hypospadias in children.Methods The 164 children with hypospadias analyzed retrospectively were treated from February,2013 to February,2021.They were divided into two groups according to the operation method.82 patients in tip group were treated with tip and 82 patients in OIF group were treated with OIF.The treatment effects of the two groups were compared.Then they were divided into two groups according to the cure condition,namely,the good prognosis group(cured by operation,151 case)and the poor prognosis group(not cured by operation,13 case).Independent factors affecting the prognosis of children were analyzed by binary logistic regression.Results The cure rate of OIF group was 96.34%,which was higher than that of the tip group(87.8%),and the incidence of postoperative complications in OIF group was 7.32%,which was lower than that of the tip group(23.17%,P<0.05).The operation time in tip group(95.95±12.35)min,which was shorter than that of the OIF group(P<0.05).At the same time,the binary logistic regression analysis showed that the degree of hypospadias and the classification of barcat were the prognostic factors of children with hypospadias.The degree of penis bending,the width of penis head and the method of operation were suspicious factors.Conclusion OIF and tip have good effects in the treatment of hypospadias in children.OIF has a higher success rate,tip has a shorter operation time and fewer postoperative complications.However,the independent factors affecting the prognosis of children are preoperative hypospadias classification and barcat classification.This operation method is not an independent factor affecting the prognosis.
5.Selective adsorption of multi-walled carbon nanotubes with liquiritin and isoliquiritin.
Bo HAN ; Qing-nuan LI ; Sheng-wei WU ; Jun-gang LI ; Wen CHEN ; Wen-xin LI
Acta Pharmaceutica Sinica 2007;42(11):1222-1226
Selective adsorption of active ingredients liquiritin and isoliquiritin from Glycyrrhiza uralensis Fisch with multi-walled carbon nanotubes (MWNTs) has been studied. Distribution coefficients of liquiritin between ethanol solvent and r-MWNTs or o-MWNTs in 293K is 37.5 and 43.3, while the distribution coefficients of isoliquiritin between ethanol solvent and r-MWNTs or o-MWNTs is 717 and 1080, respectively. It was indicated that the distribution coefficient of isoliquiritin adsorbed by MWNTs was much larger than that of liquiritin, and oxidation treatment of MWNTs could obviously enhance their adsorption ability. The possible reasons that MWNTs can selectively adsorb isoliquiritin other than liquiritin were discussed on the bases of molecular structure of the active ingredients and their interaction with nanotubes.
Adsorption
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Chalcone
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analogs & derivatives
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chemistry
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isolation & purification
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Flavanones
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chemistry
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isolation & purification
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Glucosides
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chemistry
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isolation & purification
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Glycyrrhiza uralensis
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chemistry
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Nanotubes, Carbon
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chemistry
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Plants, Medicinal
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chemistry
6.Angiogenesis function of astragaloside IV in rats with myocardial infarction via PKD1-HDAC5-VEGF pathway
Lei YANG ; Nuan LIU ; Wei ZHAO ; Xing LI ; Qian WANG ; Li HAN ; Wen-Jing QI ; Yan-Ke WANG ; Bing-Yu MAO
Chinese Journal of Pharmacology and Toxicology 2018;32(4):323-324
OBJECTIVE This study aimed to investigate the role and mechanism of Astragaloside IV (AS-IV)in rats with myocardial infarction.METHODS The myocardial infarction model was established by ligation of the left anterior descending artery. The rats were randomly divided into sham, DMSO, model group, AS-IV and CID755673 groups. The rats were sacrificed 4 weeks later, and segmental heart samples were used for hematoxylin and eosin staining and masson staining. The expression of PKD1, HDAC5 and VEGF were analyzed using immunohistochemistry, reverse transcription poly-merase chain reaction and western blot. RESULTS Compared with the sham operation and DMSO groups,morphology of myocardium in model group was disordered,accompanied with necrotic myocar-dial cells and obvious collagen tissues. After treatment with AS-IV, the morphology of myocardium was obviously improved, and the number of new blood vessels increased significantly. However, after treatment with CID755673, the myocardial tissue of rats became disordered again, the necrotic cells increased, and some vessels closed. The expression levels of PKD1, HDAC5 and VEGF mRNA and protein in myocardial tissue of model group were significantly lower than the other four groups(P<0.05), whereas these levels in the AS-IV group were significantly higher than those in the other four groups (P<0.01). Additionally, the CID755673 group had significantly higher levels of PKD1, HDAC5 and VEGF mRNA and protein than the sham group, DMSO group and model group (P<0.05). CONCLUSION AS-IV may partly promote the angiogenesis of myocardial tissue in rats with myocardial infarction via the PKD1-HDAC5-VEGF pathway.
7.Viral Contamination Source in Clinical Microbiology Laboratory.
Xin Ling WANG ; Juan SONG ; Qin Qin SONG ; Jie YU ; Xiao Nuan LUO ; Gui Zhen WU ; Jun HAN
Biomedical and Environmental Sciences 2016;29(8):609-611
To understand the potential causes of laboratory-acquired infections and to provide possible solutions that would protect laboratory personnel, samples from a viral laboratory were screened to determine the main sources of contamination with six subtypes of Rhinovirus. Rhinovirus contamination was found in the gloves, cuffs of protective wear, inner surface of biological safety cabinet (BSC) windows, and trash handles. Remarkably, high contamination was found on the inner walls of the centrifuge and the inner surface of centrifuge tube casing in the rotor. Spilling infectious medium on the surface of centrifuge tubes was found to contribute to contamination of centrifuge surfaces. Exposure to sodium hypochlorite containing no less than 0.2 g/L available chlorine decontaminated the surface of the centrifuge tubes from Rhinovirus after 2 min.
Equipment Contamination
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statistics & numerical data
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Humans
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Laboratories, Hospital
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manpower
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standards
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statistics & numerical data
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Occupational Exposure
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analysis
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statistics & numerical data
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Virus Diseases
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virology
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Viruses
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genetics
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growth & development
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isolation & purification
9.Coxsackievirus B3 Infection Triggers Autophagy through 3 Pathways of Endoplasmic Reticulum Stress.
Xiao Nuan LUO ; Hai Lan YAO ; Juan SONG ; Qin Qin SONG ; Bing Tian SHI ; Dong XIA ; Jun HAN
Biomedical and Environmental Sciences 2018;31(12):867-875
OBJECTIVE:
Autophagy is a highly conserved intracellular degradation pathway. Many picornaviruses induce autophagy to benefit viral replication, but an understanding of how autophagy occurs remains incomplete. In this study, we explored whether coxsackievirus B3 (CVB3) infection induced autophagy through endoplasmic reticulum (ER) stress.
METHODS:
In CVB3-infected HeLa cells, the specific molecules of ER stress and autophagy were detected using Western blotting, reverse transcription polymerase chain reaction (RT-PCR), and confocal microscopy. Then PKR-like ER protein kinase (PERK) inhibitor, inositol-requiring protein-1 (IRE1) inhibitor, or activating transcription factor-6 (ATF6) inhibitor worked on CVB3-infected cells, their effect on autophagy was assessed by Western blotting for detecting microtubule-associated protein light chain 3 (LC3).
RESULTS:
CVB3 infection induced ER stress, and ER stress sensors PERK/eIF2α, IRE1/XBP1, and ATF6 were activated. CVB3 infection increased the accumulation of green fluorescent protein (GFP)-LC3 punctuation and induced the conversion from LC3-I to phosphatidylethanolamine-conjugated LC3-1 (LC3-II). CVB3 infection still decreased the expression of mammalian target of rapamycin (mTOR) and p-mTOR. Inhibition of PERK, IRE1, or ATF6 significantly decreased the ratio of LC3-II to LC3-I in CVB3-infected HeLa cells.
CONCLUSION
CVB3 infection induced autophagy through ER stress in HeLa cells, and PERK, IRE1, and ATF6a pathways participated in the regulation of autophagy. Our data suggested that ER stress may inhibit mTOR signaling pathway to induce autophagy during CVB3 infection.
Activating Transcription Factor 6
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metabolism
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Autophagy
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Coxsackievirus Infections
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metabolism
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Endoplasmic Reticulum Stress
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Endoribonucleases
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metabolism
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Enterovirus B, Human
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HeLa Cells
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Humans
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Protein-Serine-Threonine Kinases
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metabolism
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Signal Transduction
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eIF-2 Kinase
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metabolism
10.Exploration of IRES Elements within the ORF of the Coxsackievirus B3 Genome.
Qin Qin SONG ; Xiao Nuan LUO ; Bing Tian SHI ; Mi LIU ; Juan SONG ; Dong XIA ; Zhi Qiang XIA ; Wen Jun WANG ; Hai Lan YAO ; Jun HAN
Biomedical and Environmental Sciences 2022;35(4):322-333
Objective:
This study aimed to identify internal ribosome entry sites (IRESs) in the open reading frame (ORF) of the Coxsackievirus B3 (CVB3) genome.
Methods:
The sequences of P1, P2, or P3 of the CVB3 genome or the truncated sequences from each antithymocyte globulin (ATG) to the end of the P1, P2, or P3 gene were inserted into the pEGFP-N1 vector. After transfection, possible IRES-dependent green fluorescent protein (GFP)-fused proteins were detected by anti-GFP western blotting. The sequences of possible IRESs were inserted into specific Fluc/Rluc bicistronic vectors, in which the potential IRESs were determined according to the Fluc/Rluc activity ratio. Expression of Fluc and Rluc mRNA of the bicistronic vector was detected by RT-qPCR.
Results:
After transfection of full length or truncated sequences of the P1, P2, or P3 plasmids, six GFP-fused protein bands in P1, six bands in P2 and nine bands in P3 were detected through western blotting. Two IRESs in VP2 (1461-1646 nt) and VP1 (2784-2983 nt) of P1; one IRES in 2C (4119-4564 nt) of P2; and two IRESs in 3C (5634-5834 nt) and 3D (6870-7087 nt) of P3 were identified according to Fluc/Rluc activity ratio. The cryptic promoter was also excluded by RT-qPCR.
Conclusion
Five IRESs are present in the CVB3 coding region.
Internal Ribosome Entry Sites/genetics*
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Open Reading Frames
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RNA, Messenger/genetics*