1.Study on the expression of a substrate-based polypeptide and its inhibition against HCV protease in vitro
Wen LIU ; Wei HU ; Shigan LING
Chinese Journal of Microbiology and Immunology 2008;28(2):119-122
Objective To synthesize an inhibitory peptide against serine protease of hepatitis C virus(HCV)and explore its inhibition ability in vitro. Methods Based on the sequence characteristics of four natural substrates of protease in HCV,the gene sequence of an inhibitory peptide against HCV serine protease was designed and directly synthesized by PCR. The segment was subcloned into prokaryotic expression vector pBVIL1,resulting in the construction of the recombinant plasmid pBVIL1/IP,which was then transformed into E. coli HB101 strain. Purified by ion exchange chromatography,the expressed protein was added into an in vitro system,which was comprised of the inhibitory peptide(expressed protein),protease and the substrate,i.e. a NS5A-B fragment,in phosphate buffer. Then SDS-PAGE was performed to test the inhibition effect of the polypeptide. Results The recombinant expression vector pBVIL1/IP containing target gene was successfully constructed. The peptide expressed as inclusion boay was identified by SDS-PAGE. The degradation of protease substrate NS5A-B fragment is inhibited proportionally with the increasing concentrations of the peptide. Conclusion The recombinant peptide shows inhibitory effect on HCV protease in vitro.
2.Preliminary Study of Magnetic Resonance T1ρ in Early Detection of Disc Degeneration
Wei WANG ; Wen LIANG ; Ling CHEN ; Jianming YANG ; Xianyue QUAN
Chinese Journal of Medical Imaging 2013;(6):406-410
Purpose To investigate the value of 3.0T MR T1ρquantitative analysis in early detection of disc degeneration. Materials and Methods Conventional T2WI and T1ρwere collected in 35 healthy volunteers (male 16, female 19) on 3.0T MRI, and the classification of disc nucleus was performed by Pfirrmann classification. T1ρvalue of nucleus pulposus was measured and analyzed for the relationship among Pfirrmann classification, segmental and gender. Results There was a significant negative correlation between T1ρvalue and Pfirrmann grade (r=-0.542, P<0.001). T1ρvalue in L5/S1 segment (94.80±26.60) ms was significantly lower than that in L2/L3 (117.18±25.64) ms and L3/L4 (115.52±28.53) ms (P<0.01), and there was no significant difference among other segments (P>0.05);there was no gender differences among the T1ρvalue of each segment (t=0.006, 0.042, 0.797, 1.022, 0.038, P>0.05). Conclusion T1ρ value is closely related to the degree of disc degeneration, and T1ρimaging can be used as an objective, sensitive and effective tool for early detection of disc degeneration.
3.Research on Deep-sea Microbiology
Chang-Hui ZHAO ; De-Zan YE ; Wen-Ling WEI ;
Microbiology 1992;0(03):-
Nutritional supplies & biodiversities of deep-sea microorganisms and correlative methodologies were introduced here in an ecological point of view, and development of their natural products was prospective in an applied point of view.
4.Modification by wheat germ agglutinin delays the ocular elimination of liposome.
Wen-Jian ZHANG ; Dong-Xiao YANG ; Ling-Lin FENG ; Fei WANG ; Gang WEI ; Wei-Yue LU
Acta Pharmaceutica Sinica 2014;49(4):543-549
The purpose of this study is to explore the feasibility of wheat germ agglutinin (WGA) modified liposome as a vehicle for ophthalmic administration. Liposome loaded with 5-carboxyfluorescein (FAM) was prepared by lipid film hydration method. WGA was thiolated and then conjugated to the surface of the liposome via polyethylene glycol linker to constitute the WGA-modified and FAM-loaded liposome (WGA-LS/FAM). The amount of thiol groups on each WGA molecule was determined, and the bioactivity of WGA was estimated after it was modified to the surface of liposome. The physical and chemical features of the WGA-modified liposome were characterized and the ocular bioadhesive performance was evaluated in rats. The result showed that each thiolated WGA molecule was conjugated with 1.32 thiol groups. WGA-LS/FAM had a mean size of (97.40 +/- 1.39) nm, with a polydispersity index of 0.23 +/- 0.01. The entrapment efficacy of FAM was about (2.95 +/- 0.21)%, and only 4% of FAM leaked out of the liposome in 24 h. Erythrocyte agglutination test indicated that after modification WGA preserved the binding activity to glycoprotein. The in vivo ocular elimination of WGA-LS/FAM fitted first-order kinetics, and the elimination rate was significantly slower than that of the unmodified liposome, demonstrating WGA-modified liposome is bioadhesive and suitable for ophthalmic administration.
Absorption, Physicochemical
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Adhesiveness
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Administration, Ophthalmic
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Animals
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Drug Carriers
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Eye
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metabolism
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Fluoresceins
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chemistry
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Liposomes
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administration & dosage
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chemistry
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pharmacokinetics
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Male
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Particle Size
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Polyethylene Glycols
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chemistry
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Rats
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Rats, Sprague-Dawley
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Wheat Germ Agglutinins
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administration & dosage
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chemistry
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pharmacokinetics
5.The curative effects of transmetil on Amanita verna poisoning.
Ling WEN ; Wei-wei LIU ; Jian-wei HUANG ; Wei YU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2011;29(2):134-136
OBJECTIVETo observe the curative effects of transmetil on Amanita verna poisoning.
METHODSTwelve cases with Amanita verna poisoning were reviewed. The patients were divided into 2 groups according to usage of transmetil: Group A was treated with traditional protocol (gastric lavage, catharsis, rehydration, diuresis, anti-infection and hemodialysis), Group B was treated with traditional protocol combined with transmetil. The liver function changes on the 1st, 3rd, 5th and 7th day after poisoning and the mortality were compared between 2 groups.
RESULTSTwo cases in group A (6 patients) died. The mortality of group A was 33.3%. The AST levels continued to increase on the 3rd and 5th day, but decreased on the 7th day. TBIL continued to increased on the 1st, 3rd, 5th and 7th day. None in group B died. The TBIL level dropped at 7 d 5 patients showed an increase in ALT at 7 d and 3 patients showed a decrease in AST at 7 d.
CONCLUSIONTransmetil may play an important role in reducing the mortality of Amanita verna poisoning.
Adolescent ; Adult ; Aged, 80 and over ; Amanita ; Female ; Humans ; Male ; Middle Aged ; Mushroom Poisoning ; drug therapy ; Retrospective Studies ; S-Adenosylmethionine ; therapeutic use
6.Quality analysis of Guizhi Fuling capsule before and after application of in-process quality control in pharmaceutical production.
Jia-chun LI ; Jin-ling WANG ; Jing-ling WU ; Wen-zhe HUANG ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(6):1017-1022
The effects of application of in-process quality control in Guizhi Fuling capsule production were evaluated by 192 batches data analysis. Using a statistical analysis method, each batch of data were to be counted to research for the difference between 96 samples adopting the technologies of in-process control or not. According to quality standards of Guizhi Fuling capsule, all measurements of the 192 batches of the drugs before and after the application of process control technology were analyzed, and they were within the rules. There was a significant difference between adopting the technologies of process control or not. Application of in-process control technology can improve the uniformity of lot-to-lot for Guizhi Fuling capsule.
Capsules
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chemistry
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Chemistry, Pharmaceutical
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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chemistry
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Quality Control
7.Clinical efficacy observation of acupoint thread-embedding in treating obese patients with food addiction
Ling-Ling JI ; Ba-Si OUYANG ; Jie SHEN ; Ying LUO ; Ya-Wen JIANG ; Bing-Wei AI ; Jue HONG
Journal of Acupuncture and Tuina Science 2021;19(3):193-199
Objective: To observe the effects of acupoint thread-embedding therapy and low-carbohydrate diet therapy on obese patients with food addiction. Methods: Sixty-five eligible patients were randomized into a thread-embedding group of 33 cases and a diet group of 32 cases to respectively receive 12-week treatment. Before treatment, after treatment and at 6-month follow-up, the two groups were observed and compared in terms of body mass (BM), waist circumference (WC), hip circumference (HC), waist-to-hip ratio (WHR), body mass index (BMI), body fat rate (BFR), basal metabolic rate (BMR) and Yale food addiction scale version 2.0 (YFAS 2.0). Results: At the end of treatment, there were no significant differences in the general efficacy, and the improvements in BM, BMI, WC, HC, WHR and BFR between the thread-embedding group and diet group (all P>0.05). At follow-up, the thread-embedding group showed more significant improvements in all the aforementioned indicators compared with the diet group except HC (all P<0.05). At the end of treatment and follow-up, BMR and YFSA 2.0 had more significant improvements in the thread-embedding group than in the diet group (all P<0.05). Conclusion: Acupoint thread-embedding therapy can produce significant efficacy in treating obese patients with food addiction; it can improve the food addiction state and work better in maintaining the efficacy compared with low-carbohydrate diet therapy.
8.Synthesis and biodistribution of a bi-functional agent 99Tcm-Gd-DTPA-DG in tumor bearing nude mice
Wei, ZHANG ; Liang, CAI ; Yue, CHEN ; Zhan-wen, HUANG ; Zhi-ling, DING ; Feng, CAO ; Li, ZHANG ; Ling, HE
Chinese Journal of Nuclear Medicine 2011;31(2):117-120
Objective To evaluate the stability and biodistribution of a novel SPECT-MRI bi-functional agem 99Tcm-Gd-DTPA-DG in tumor-bearing nude mice. Methods DTPA-DG was synthesized and then conjugated with Gd2O3 to generate Gd-DTPA-DG. The tumor-bearing nude mice were scanned by MRI to evaluate the tumor targeting ability of Gd-DTPA-DG. The orthogonal experiment was applied to optimize pH value of reaction medium and reaction temperature. The radiolabeling efficiency was measured by thin layer chromatography. The distribution of 99Tcm-Gd-DTPA-DG in nude mice was evaluated by scintigrapy in vivo. The % ID/g was measured at different time after intravenous injection of 99Tcm-Gd-DTPA-DG. Results The tumor was significantly enhanced by Gd-DTPA-DG with MRI. The radiochemical purity of 99Tcm-Gd-DTPADG was about 98.5% and remained 96.2% at room temperature for 6 h. The tumor was well visualized by 99TcmGd-DTPA-DG SPECT at 2 h after injection. The tumor uptake was (1.48 ±0.12) %ID/g, and the rumor to muscle radioactivity ratio was 2.91. Conclusions MRI contrast of Gd-DTPA-DG may enhance tumor detection. 99Tcm-labeled Gd-DTPA-DG may be useful for tumor imaging and might have a potential role as a SPECT-MRI bi-functional agent.
9.Correlation between UGT1A1 Polymorphism and Neonatal Hyperbilirubinemia of Neonates in Wuhan
LIU WEI ; CHANG LI-WEN ; XIE MIN ; LI WEN-BIN ; RONG ZHI-HUI ; WU LI ; CHEN LING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(5):740-743
This study attempts to discuss the correlation between UGT1A1*28 as uridine diphosphate glucuronosyltransferase gene promoter and coding region Gly71Arg gene polymorphism with neonatal hyperbilirubinemia of neonates in Wuhan.A total of 168 neonates were divided into the hyperbilirubinemia group (case group,n=108) and healthy neonates group (control group,n=60).Their DNA was obtained through blood extraction.The gene exon mutation of UGT1A1 was detected by Sanger sequencing,which revealed the relationship between UGT 1A 1*28 and Gly71Arg polymorphism with neonatal hyperbilirubinemia of neonates.The results showed that:(1) The frequency of UGT1Al*28 allele mutation in the case group and the control group was 9.3% and 10% respectively,with the difference being not significant between the two groups (P>0.05).(2) The frequency of Gly71Arg allele mutation in the case group and the control group was 35.1% and 21.7% respectively,with the difference being significant between the two groups (P<0.01).(3) The serum bilirubin level of Gly71Arg mutant homozygous and heterozygous subgroups (n=66) in the case group was 302.7±31.4 μmol/L,which was significantly higher than 267.3±28.5 μmol/L of the wild subgroup (n=42) (P<0.01).It was suggested that the occurrence of neonatal hyperbilirubinemia of neonates in Wuhan was not associated with UGT 1A1*28 gene polymorphism,but closely with the Gly71Arg gene polymorphism.Meanwhile,the Arg allele mutation was related to the degree of jaundice.
10.Effects of RNA binding protein RNPC1 on biological function of renal cell carcinoma cells
Wen HUANG ; Chunmei JI ; Haiwei YANG ; Liang SHI ; Ling MENG ; Jifu WEI
Journal of Medical Postgraduates 2017;30(4):365-370
Objective RNPC1 may act as an oncogene or suppressor gene in human tumors and its role in human renal cell carcinoma (RCC) remains unclear.The objective of this study was to investigate the role of RNPC1 in the development of RCC.Methods Over-expression of RNPC1 gene group (RNPC1 group) and short hairpin RNA interfering RNPC1 gene expression (shRNPC1 group) were respectively built in RCC CAKI-1 and CAKI-2.The blank control group (NC group) and negative control group (SCR group) were built as well.The qRT-PCR and western blot (WB) were used to detect the expression levels of RNPC1 mRNA and RNPC1 protein in RCC cells.Lentivirus infection was applied to establish stable expressed RCC cell lines of RNPC1 over-expression and interference.Detection was made on mRNA and protein expression levels in RNPC1 stable RCC cell lines.The effects of RNPC1 on cell proliferation, colony formation assay, migration, and invasion were detected by CCK-8 cell differentiation test, clone test, scratch test, and migration and invasion test.WB was applied to detect the change of protein expression in the EMT path of RNPC1 stable RCC cell lines and explore the molecular mechanism of RNPC1 effect on the biological function of RCC cells.Results The expression levels of RNPC1 mRNA and protein were found lower in shRNPC1 group than those in SCR group, while the expression levels of RNPC1 mRNA and protein in SCR group were higher than those NC group (P<0.05).The capability of proliferation in shRNPC1 group was stronger than that in SCR group, while the capability of proliferation in shRNPC1 group was weaker than that in NC group (P<0.05).The capabilities of cell migration and invasion were stronger in shRNPC1 group than those in SCR group, while the capabilities of cell migration and invasion in RNPC1 group were weaker than those in NC group (P<0.05).RNPC1 could inhibit the proliferation capability of RCC cells and might up-regulate the protein expression of E-cadherin and down-regulate the protein expression of β-catenin and vimentin, thus inhibiting EMT path and the capabilities of migration and invasion off RCC cells (P<0.05).Conclusion RNPC1 acts as a tumor suppressor in RCC and has the potential for the prediction of RCC prognosis.