1.Characterization of Proteins and Nucleic Acid of Rice Stripe Virus
Hanxin, LIN ; Qitian, LIN ; Zujian, WU ; Qiying, LIN ; Lianhui, XIE
Virologica Sinica 1999;14(4):343-352
An isolate of rice stripe virus (designated as RSV-YL) was purified. The particles showed to be pleomorphisms under electron microscope, mainly branched filaments of about 80-250 nm in length and about 8 nm in width. There are also some open circular filaments of 3 nm and 8 nm in width, and some filaments of 13 nm in width and 130-190 nm in length. The basic morphism of RSV particles should be filaments of 3 nm in width and various length. By SDS-PAGE analysis, the molecular weight of disease-specific protein (SP) encoded by vRNA4 was 19.9 kDa and that of coat protein (CP) encoded by vcRNA3 was 33.6 kDa. When nucleic acid extracted from the purified RSV was electrophoresed under nondenaturing condition, the size of four dsRNAs (designated as dsRNA1-4 in order of decreasing size) was 4.9×106,2.7×106,2.0×106 and 1.7×106 Da, respectively, and that of four ssRNAs (designated as ssRNA1-4 in order of decreasing size) was 3.0×106,1.2×106,0.9×106 and 0.8×106 Da, respectively. A fifth segment with a size of 0.58×106 Da identified as ssRNA5 associated with the purified virus sometimes. The antiserum against the coat protein further purified by preparative electrophoresis was raised and used to investigate the serological relationships between RSV-CP and RSV-SP, CP and SP of rice grassy stunt virus (RGSV) which is also a member of Tenuivirus. The results showed that RSV-CP had no serological reaction with SP of RSV and PGSV, but could weakly react with antiserum of RGSV-CP, which confirmed that there is distantly evolutionary relationship between RGSV and RSV.
2.Characterization and Amino Acid Sequence of y3,an Antiviral Protein from Mushroom Coprinus comatus
Liping WU ; Zujian WU ; Dong LIN ; Fang FANG ; Qiying LIN ; Lianhui XIE
Chinese Journal of Biochemistry and Molecular Biology 2008;24(7):597-603
Many proteins from mushrooms can be used as potential antiviral agents and in plant protection.An antiviral protein,designated as y3,Was isolated from fruiting bodies of the fungus Coprinus comatus by fastflow ion-exchange column chromatography coupled with high-resolution molecular sieve chromatography.This glycoprotein was detected in both fruiting body and mycelium by Western blot analysis.According to its Nterminal sequence.an amino acid sequence and a partial cDNA sequence of the y3 gene were obtained.Protein y3 at 2.0μg/ml achieved 50.0% inhibition of tobacco mosaic virus(TMV,20μg/ml)lesions in Nicotiana glutinosa leaves.It also inhibited multiplication of TMV in Nicotiana tabacum Var.K326.
3.Expression of Rice Gall Dwarf Virus Outer Coat Protein Gene (S8) in Insect Cells
Guocheng FAN ; Fangluan GAO ; Taiyun WEI ; Meiying HUANG ; Liyan XIE ; Zujian WU ; Qiying LIN ; Lianhui XIE
Virologica Sinica 2010;25(6):401-408
To obtain the P8 protein of Rice gall dwarf virus (RGDV) with biological activity, its outer coat protein gene S8 was expressed in Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression system. The S8 gene was subcloned into the pFastBacTM1 vector, to produce the recombinant baculovirus transfer vector pFB-S8. After transformation, pFB-S8 was introduced into the competent cells (E. coli DH10Bac) containing a shuttle vector, Bacmid, generating the recombinant bacmid rbpFB-S8. After being infected by recombinant baculovirus rvpFB-S8 at different multiplicities of infection, Sf9 cells were collected at different times and analyzed by SDS-PAGE, Western blotting and immunofluorescence microscopy. The expression level of the P8 protein was highest between 48-72 h after transfection of Sf9 cells. Immunofluorescence microscopy showed that P8 protein of RGDV formed punctate structures in the cytoplasm of Sf9 cells.
4.Therapeutic value of endoscopic resection of tumors originated from gastric fundus muscularis propria
Qiuyong XU ; Junjie CHEN ; Yadong LAI ; Hui PU ; Qiying LIU ; Shuhui LIN ; Xuefen CHEN ; Xiangnong XU
Chinese Journal of Digestive Endoscopy 2015;(3):175-179
Objective To investigate the safety and effectiveness of endoscopic resection of tumors originated from gastric fundus muscularis propria.Methods Data of 53 patients with tumors originated from gastric fundus muscularis propria detected by endoscopic ultrasonograpy,treated by endoscopic resection and followed up at our hospital between January 2012 and June 2014 were reviewed.The postoperative pathology and complications were retrospectively analyzed to evaluate the therapeutic effect and safety.Results The procedure was successfully performed on all patients and all lesions were removed in one procedure.The lesion size ranged from 0.5 to 4.5 cm and the operation time was 25-155 min[mean(46.7 ±18.2)min].Mild bleeding (5 ~150 ml)occurred in all cases,which was successfully managed by argon plasma coagulation,hot biopsy probe or endoclip.Perforation occurred in 8 patients(8 /53),seven of whom were closed with titanium clips and titanium clips combined with nylon cord.Laparoscopic intervention was applied to 1 case because of severe perforation.Gastrointestinal decompression,acid suppression with proton pump inhibitors and antibiotics were performed on all cases.No severe hemorrhage occurred.The average length of hospitalization was (5.3 ± 1.4)days(3-14 d).Pathology confirmed 46 cases of gastrointestinal stromal tumors and 7 cases of leiomyoma. The patients were followed up for 3 to 27 months,and no tumor residue or recurrence was observed. Conclusion Endoscopic resection is a method not only to get the accurate pathologic diagnosis but also to meet principle of the local resection for stomach.It is safe,effective and worthy of recommendation.
6.Exploration of risk factors and establishment of nomograms model for postoperative adjuvant chemotherapy in stage Ⅰ gastric cancer
Li LI ; Yunhe GAO ; Benlong ZHANG ; Zijian WANG ; Qiying SONG ; Hao CUI ; Zhi QIAO ; Lin CHEN
International Journal of Surgery 2023;50(5):306-311,C1
Objective:To identify the risk factors associated with postoperative adjuvant chemotherapy in patients with stage I gastric cancer and establish nomograms model based on risk factors.Methods:In this retrospective case-control study, 161 cases with stage Ⅰ primary gastric adenocarcinoma were included who underwent gastrectomy at the Department of General Surgery of the First Medical Center of Chinese PLA General Hospital from January to December in 2020, including 129 male cases and 32 females cases, with the average age of (59.90±0.80) years. Among them, 41 cases were treated with postoperative adjuvant chemotherapy (chemotherapy group), while 120 cases who did not receive postoperative adjuvant chemotherapy (no chemotherapy group). Univariate and multivariate Logistic regression analyses were used to identify the risk factors of adjuvant chemotherapy in stage Ⅰ gastric cancer patients and establish the nomograms predictive model. ROC curve and calibration curve were used to evaluate the performance of the model.Results:Multivariate analysis revealed that primary tumor site, tumor size, T stage, N stage lymph-vascular tumor embolus or perineural invasion were the independent risk factors of postoperative adjuvant chemotherapy for stage Ⅰ gastric cancer( P<0.05). The ROC curve indicated that area under the curve (AUC) of the multivariate model was 0.91(95% CI: 0.86-0.97). The calibration curve showed that probability predicted by nomograms was consistent with the actual situation(C-index: 0.91). Conclusions:The tumor located in the proximal stomach, tumor size>2 cm, T 2, N 1, lymph-vascular tumor embolus or perineural invasion maybe be the risk factors for chemotherapy decision in stage Ⅰ gastric cancer patients. The established model has good predictive ability for postoperative chemotherapy of stage Ⅰ gastric cancer patients, which might provide reference for the selection of clinical decisions in this part of patients.
7.The antioxidant effect of serum bilirubin in lung cancer
Lidong LIU ; Zixing MA ; Qiying LIN ; Chengzhi ZHOU
The Journal of Practical Medicine 2018;34(10):1716-1719,1724
Objective To study the antioxidant effect of serum bilirubin,an endogenous antioxidants,in the patients with lung cancer. Methods A total of 402 patients with lung cancer(patient group)and 422 healthy individuals(control group)were enrolled in this study. The concentrations of total bilirubin(TBIL)and direct bili-rubin(DBIL)were measured using Diazonium Salts reagent on an automated chemistry analyzer(AU5821,Beck-man Coulter)and concentrations of indirect bilirubin(IBIL)were calculated based on concentrations of TBIL and DBIL. The total activity of SOD and MDA concentrations of 104 patients and 57 controls were measured using Xan-thine oxidase and TBA reagents respectively. All data were analyzed using SPSS 19.0 Software. Results Compared with the control group,the concentrations of TBIL,DBIL,IBIL and the activity of SOD in the patient group were decreased(P < 0.05)but the concentrations of MDA were increased. However,no tendency was found from T1 to T4(TNM)groups. Conclusion There is antioxidant dysfunction within patient with lung cancer and the serum bilirubin would be involved in this process.
8.Preparation of Lir@BSA-PMF nanoparticles and verification of their cell functions
Qingyu HUANG ; Qiying CHEN ; Shengjia SUN ; Bangwei WU ; Shan LIN ; MAIMAITIJIANG ALIMUJIANG
Basic & Clinical Medicine 2024;44(2):235-241
Objective To synthesize bovine serum albumin(BSA)-loaded liraqlutide(Lir)-nanoparticles coated with platelet membrane fragments(PMF)using a"bottom-up"nano-engineering chemistry technique,and to evaluate their cyto-compatibility and potential function of anti-oxidative stress.Methods PMF was extracted as reported previously.Lir@BSA nanoparticles were prepared by self-assembly method.PMF was coated on the sur-face of Lir@BSA nanoparticles by co-extrusion to prepare Lir@BSA-PMF.The physical and chemical properties of Lir@BSA-PMF particles were characterized as particle size,Zeta potential,transmission electron microscopy and particle size stability.The encapsulation efficiency,loading efficiency and cumulative release efficiency of liraglu-tide were calculated by enzyme-linked immunosorbent assay.Further,SDS-PAGE was used to analyze whether there was a similar membrane protein distribution of platelet membrane on Lir@BSA-PMF bionicnanocarrier.CCK-8 assay was used to verify the biocompatibility of the materials.Reactive oxygen species(ROS)experi-ment was used to explore the effect of Lir@BSA-PMF on cell oxidative damage.The uptake of cells on Lir@BSA-PMF bionic nano capsules was verified by cell phagocytosis experiment.Results Lir@BSA-PMF nanop-articles had a stable particle size of 25 nm with a spherical morphology,and a Zeta potential value of-25.5 mV.The encapsulation efficiency,loading efficiency and cumulative release efficiency of liraglutide were 85.56%,7.96%and 77.06%,respectively.SDS-PAGE analysis showed that the Lir@BSA-PMF bio-mimetic nano capsules retained the similar membrane protein distribution as platelet membrane.CCK-8 assay verified that the nanomaterials were non-cytotoxic.ROS results showed that Lir@BSA-PMF nanomaterials had obvious antioxidant properties.The results of cell phagocytosis showed that the cells had a good phagocytosis effect on Lir@BSA-PMF nanoparticles.Conclusions The nanoparticles Lir@BSA-PMF are successfully syn-thesized and have no effects on cells viability in vitro.The particles are taken up by cells and show a significant function of antioxidant damage.
9.NOTCH3 gene mutations in two Chinese families featuring cerebral autosomal dominant arteriopathy with subcortical infarct and leucoencephalopathy.
Qiying SUN ; Wenwen LI ; Yafang ZHOU ; Fang YI ; Jianfeng WANG ; Yacen HU ; Lingyan YAO ; Lin ZHOU ; Hongwei XU
Chinese Journal of Medical Genetics 2017;34(6):816-820
OBJECTIVETo analyze potential mutations of the NOTCH3 gene in two Chinese families featuring cerebral autosomal dominant arteriopathy with subcortical infarct and leucoencephalopathy (CADASIL).
METHODSThe two probands and related family members and 100 healthy controls were recruited. Potential mutations of the NOTCH3 gene were screened by PCR and direct sequencing. PolyPhen-2 and SIFT software were used to predict the protein function.
RESULTSThe conditions of both probands were adult-onset, with main clinical features including recurrent transient ischemic attacks and/or strokes, cognitive impairment. MRI findings suggested multiple cerebral infarcts and severe leukoencephalopathy. A heterozygous mutation c.328C>T (p.Arg110Cys), which was located in exon 3 of the NOTCH3 gene and known as a causative mutation, was identified in proband 1. A novel heterozygous mutation c.1013 G>C (p.Cys338Ser) located in exon 6 of the NOTCH3 gene was identified in the proband 2, which was not reported previously. The same mutations were not detected among the 100 unrelated healthy controls. Function analysis suggested that heterozygous mutation c.1013G>C can severely affect the functions of NOTCH3 protein.
CONCLUSIONTwo heterozygous missense mutations in the NOTCH3 gene have been identified in two families affected with CADASIL. The novel heterozygous Cys338Ser mutation in exon 6 of the NOTCH3 gene probably underlies the CADASIL.
Adult ; Brain ; diagnostic imaging ; CADASIL ; diagnostic imaging ; genetics ; Female ; Heterozygote ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Mutation ; Receptor, Notch3 ; genetics
10.Survey of related factors of maternal venous thromboembolism in nine hospitals of China
Zilian WANG ; Huizhen GENG ; Xianlan ZHAO ; Qiying ZHU ; Jianhua LIN ; Li ZOU ; Yang MI ; Yali HU ; Shangrong FAN ; Xu CHEN ; Zhe LIU ; Huixia YANG
Chinese Journal of Obstetrics and Gynecology 2020;55(10):667-672
Objective:To investigate and analyze disease status and risk factors of venous thromboembolism (VTE) during pregnancy and puerperium in our country.Methods:Clinical datas were collected from 575 patients diagnosed with VTE during pregnancy and puerperium and hospitalized in nine medical institutions in our country from January 1, 2015 to November 30, 2019, and retrospectively analyzed it′s disease status and risk factors.Results:(1) The proportion of VTE in pregnancy and puerperium was 50.6% (291/575) and 49.4% (284/575), respectively. Four patients died, the mortality rate was 0.7% (4/575). The cause of death was pulmonary embolism. (2) The location of VTE during pregnancy and puerperium was mainly in the lower limb vascular (76.2%, 438/575), followed by pulmonary vessels (7.1%, 41/575). (3) In the risk factors of VTE, cesarean section accounted for 32.3% (186/575), maternal advance age accounted for 27.7% (159/575), braking or hospitalization during pregnancy accounted for 13.6% (78/575), other risk factors accounted for more than 5% were previous VTE, obesity, preterm birth, assistant reproductive technology conception and so on, pre-eclampsia and multiple pregnancy accounted for 4.9% (28/575) respectively. In addition, some patients with VTE did not have any of the above risk factors, and the incidence rate was as high as 23.1% (133/575).Conclusions:The occurrence of VTE during pregnancy and puerperium is related to multiple risk factors, and could lead to matemal death, It is very necessary to screen VTE risk factors for all pregnant women, to make corresponding prevention and control measures.