1.Cloning of full-length cDNA of Microsporum canis membrane protein PQ-loop repeat protein gene
Juan PANG ; Yiping ZHU ; Guoling YANG
Chinese Journal of Dermatology 2012;45(2):138-139
ObjectiveTo clone the full-length cDNA of Microsporum canis membrane protein PQ-loop repeat protein(PQ-LRP) gene,so as to investigate the roles of PQ-LRP in the pathogenesis of tinea capitis.MethodsA Microsporum canis strain(A518) from a patient with tinea capitis served as the experimental strain.Rapid cDNA end amplification(RACE) was performed to clone the full length cDNA sequence of PQLRP gene.Bioinformatics methods were used to make a preliminary functional analysis of the gene.Results The cDNA of PQ-LRP gene was obtained with a full length of 1522 bp,including the 5' untranslated region (49 bp),coding region(1080 bp) and 3' untranslated region(393 bp).The coding region encoded a protein precursor including 359 amino acid residues.The cloned cDNA of PQ-LRP gene shared an 81% nucleotide identity with that of Trichophyton tonsurans and a 79% nucleotide identity with that of Trichophyton rubrum.Conclusions The full-lengthcDNA of Microsporumcanis membraneproteinPQ-LRP gene hasbeen successfully cloned,which will provide an important basis for further researches into the roles of PQ-LRP in Microsporum canis-associated diseases.
3.The Change of the Level of Plasma D-dimer in Coronary Heart Disease.
Xinquan PANG ; Wenge NIU ; Juan KONG
Journal of Medical Research 2006;0(02):-
Objective To study the change of the level of plasma D-dimer(DD) in patients with coronary heart disease(CHD)and its significance.Methods 96 CHD patients were divided into various groups according to coronary angiography and clinical manifestation: 26 patients with simple lesions,47 patients with complex lesions,12 patienty with acute myocardial in-farction(AMI),42 patienty with unstable angina pectoris(UAP),19 patienty with stable angina pectoris(SAP)and 23 patienty with normal healthy control subjects.The level of plasma DD was detected.Results Plasma concentrations of DD were higher in patients with complex lesion((0.501?0.209)mg/L)than in those with simple lesions((0.328?0.1)mg/L)(P
4.Isolation and structure identification of steroidal saponin from Dioscorea zingiberensis
Juan CHENG ; Changying HU ; Zijie PANG ; Deping XU
Chinese Traditional and Herbal Drugs 1994;0(02):-
Objective To study the chemical constituents of Dioscorea zingiberensis.Methods The fresh rhizome of D.zingiberensis was extracted three times,3 h once with EtOH-H2O at 80 ℃.The EtOH was evaporated under reduced pressure to give a residue which was suspended in water and then was exacted with petroleum ether,ethyl acetate,and n-butanol fraction.The water fraction was isolated by the reversed-phase ODS column chromatography.The chemical structures were elucidated by means of 1H-NMR,13C-NMR,135DEPT,HMQC,and HMBC spectroscopic analyses.Results Three steroidal saponins were isolated from the fresh rhizome of D.zingiberensis.The compounds were identified as:diosgenin-3-O-?-D-glucopyranosyl(1→3)-?-D-glucopyranosyl(1→4)-[?-L-rhamnopyranosyl(1→2)]-?-D-glucopyranoside(Ⅰ),(25R)-26-O-?-D-glucopyranosyl-furost-5-en-3?,22?-diol-3-O-?-D-glucopyranosyl(1→3)-?-D-glucopyranosyl(1→4)-[?-L-rhamnopyranosyl(1→2)]-?-D-glucopyranoside(Ⅱ),and(25R)-26-O-?-D-glucopyranosyl-furost-5-en-3?,22?-diol-7-carbonyl-3-O-?-D-glucopyranosyl(1→3)-?-D-glucopyranosyl(1→4)-[?-L-rhamnopyranosyl(1→2)]-?-D-glucopyranoside(Ⅲ).Conclusion Compound Ⅲ is a novel compound named as zingiberenin H.
5.Expression of FSH1 in Microsporum canis before and after the induction by skin tissue from the scalp and foreskin of children
Yiping ZHU ; Zhenying ZHANG ; Juan PANG ; Guoling YANG
Chinese Journal of Dermatology 2011;44(6):441-442
Objective To explore the significance of FSH1 in the pathogenicity of M. canis. Methods Thirty M. cam's strains from tinea capitis lesions and 30 M. canis strains from tinea corporis lesions were cultured, passaged, and induced by medium containing skin tissue of scalp or foreskin from children. Semi-quantitative reverse transcription (RT)-PCR was carried out to detect the expression of FSH1 mRNA in the firstand fifth-generation M. canis strains, as well as M. canis strains induced by the skin tissues. Results The mRNA expression of FSH1 was higher in M. canis strains derived from tinea capitis lesions than in those from tinea corporis lesions (P < 0.01), but there was no significant difference between the first-generation and fifthgeneration M. canis strains (P> 0.05). The skin tissue from scalp and foreskin induced a significant elevation in the mRNA expression of FSH1 in these M. canis strains (F = 2025.713, 1833.139, both P< 0.01), and the inductive effect of the scalp tissue was different from that of the foreskin tissue (P < 0.01). Conclusions The FSH1 mRNA expression is different in M. canis isolated from different body sites. Local skin tissue has an inductive effect on the expression of FSH1 mRNA, and the inductive effect of scalp tissue is more apparent than that of foreskin tissue.
6.Identification of genetic mutation in a Chinese pedigree with congenital cataract by whole-exome sequencing
Juan, BU ; Jing, LIU ; Honglei, PANG ; Feng, LIU ; Lejin, WANG
Chinese Journal of Experimental Ophthalmology 2016;34(10):896-899
Background Genetic mutation remains to be the most common cause of congenital cataract.Whole exon sequencing technology is an ideal method to detect the pathogenic gene mutations.Objective This study was to identify the pathogenic gene in a Chinese autosomal dominant congenital cataract (ADCC) family by whole-exome sequencing.Methods This study complied with Helsinki Declaration and the protocol was approved by Ethic Committee of Peking University Third Hospital.Informed consent was obtained from each subject before any medical examination.A cross-sectional study was designed.A Chinese ADCC family with 4 generations and 48 members were enrolled in Peking University Third Hospital,of which Ⅰ1 and Ⅰ2 died.The periphery blood of 8-10 ml was collected from each member of Ⅱ,Ⅲ and Ⅳ generations for the high throughput sequencing of genes using whole exon trapping and new sequencing technology,and the sequencing results were compared with the data of human HA PMAP8,dbSNP130 and 1000 Genome Project database.The synonymous mutation was filtered after reported common variants,and the false positive results of explicit sequencing were finally excluded by Sanger sequencing and then the candidate genes were identified.The mutation genes were screened to determine the pathogenic gene of this ADCC family.Results Eleven ADCC patients were found in this family,and the patients distributed in each generation with an equal chance for involvement in male and female subjects,which conformed to an autosomal dominant inheritance pattern.All the patients were nuclear cataract.Genome-wide whole-exome sequencing found that major intrinsic protein (MIP) gene was known genes of ADCC in initially identified candidate genes,so the Sanger was used to verify the MIP gene.The heterozygous mutation of MIP gene (chr12:56845250 C > T) appeared to be the pathogenic cause of this ADCC family.The mutation occurred in the splice sites of the gene,resulting in the fourth exon coded-61 amino acids are replaced by leucine,histidine and serine,which lead to the abnormal truncated proteins.Conclusions The heterozygous mutation of MIP gene is the molecular pathogenesis of this Chinese ADCC family.
7.Purification of superoxide dismutase from blood erythrocyte by nano magnetic chitosan microspheres.
Baoquan WANG ; Juan PING ; Feng LI ; Yongzhou ZHANG ; Cui LIU ; Xiaobin PANG
Chinese Journal of Biotechnology 2014;30(6):990-994
Nano magnetic microspheres prepared by chitosan and poly acylic acid were applied to purifying superoxide dismutase from blood erythrocyte. Chitosan-polyacyilc acid graft copolymer was synthesized by free radical graft copolymerization with potassium persulfate as inititator. To prepare Fe3O4 magnetic fluids with chemical coprecipitation, chitosan-polyacylic nano magnetic microspheres were prepared with glutaraldehyde as crosslinking agent. Structure of nano magnetic microspheres was detected by FT-IR spectrometer. Particle size and morphology were characterized by JEM-4000EX technology. Chitosan-polyacylic nanometer microspheres have good paticle cize distribution, magnetic responsiveness and protein adsoption. Activity, product yield and activity recovery of SOD after purification reached 6 727 U/mg, 21.1%, and 85.7% respectively. Purification of blood superoxide dismutase by chistosan-polyacylic acid microspheres has its renewable and feasible nature.
Chitosan
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chemistry
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Erythrocytes
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enzymology
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Glutaral
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chemistry
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Magnetics
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Microspheres
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Polymers
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Spectroscopy, Fourier Transform Infrared
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Superoxide Dismutase
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isolation & purification
8.Efficacy of intravitreal ranibizumab combined with laser photocoagulation for the treatment of macular edema induced by ischemic branch retinal vein occlusion
Recent Advances in Ophthalmology 2018;38(4):348-351
Objective To observe the therapeutic efficacy of intravitreal ranibizumab combined with laser photocoagulation for the treatment of macular edema associated with ischemic branch retinal vein occlusion (BRVO).Methods Together 60 patients (60 eyes) with BRVO in Anshan City Central Hospital were included and divided into three groups,20 patients with intravitreal ranibizumab treatment as A group,20patients with retinal laser photocoagulation treatment as B group,and another 20 patients with intravitreal ranibizumab combined with retinal laser photocoagulation therapy as C group,followed by the preoperative observation of the best corrected visual acuity (BCVA),slit lamp,fundus and intraocular pressure examination,fundus fluorescein angiography (FFA),optical coherence tomography (OCT).One day after the surgery,BCVA,slit lamp and intraocular pressure examination were conducted,and 1 month,2months,3 months after the surgery,observation of BCVA,slit lamp,fundus and intraocular pressure examination were performed.Then,OCT was used to determine the status of macular edema.And finally,BCVA and central macular thickness (CMT) in the three groups were statistically analyzed by observing the above indicators.Results The BCVA at 1 month,2 months,and 3 months after treatment was higher than before treatment in all three groups,and the differences were statistically significant (all P < 0.05).The BCVA of A group was 0.26 ±0.14 and 0.26 ±0.14 at 2 and 3 months after treatment,respectively,which was significantly better than that of B group (0.39 ±0.10 and 0.40 ±0.10) (all P <0.05).At 3 months after treatment,the BCVA in C group was 0.14 ±0.11,which was significantly higher than that in A group (0.26 ±0.14) (P<0.05).The BCVA of C group was 0.30 ±0.13,0.20 ±0.12,0.14 ± 0.11 at 1 month,2 months and 3months after treatment,respectively,which was better than that of B group (0.43 ±0.10,0.39 ± 0.10,0.40 ± 0.10),and the differences were statistically significant (all P <0.05).The postoperative CMT was significantly reduced when compared with preoperation in all three groups (all P < 0.05).The CMT at 1 month,2 months and 3 months after treatment in C group was (318.85 ± 71.48)μm,(287.15 ± 56.71) μm and (255.05 ± 60.90)μm,respectively,which was better than that in A group [(347.00 ± 67.59) μm (305.10 ± 47.44) μm and (282.40 ± 36.26) μm],and B group [(417.05 ± 63.94) μm,(394.80 ±57.18) μm,and (375.90 ± 55.10) μm],with significant differences (all P < 0.05).At 2 months and 3 months after treatment,CMT in A group was better than that in B group,and the difference was statistically significant (P < 0.05).Conclusion The efficacy of retinal laser photocoagulation combined with intravitreal ranibizumab in the treatment of BRVO macular edema is better than simple retinal laser photocoagulation and simple intravitreal ranibizumab.
9.The apoptosis induction on K562 cells by the CML bcr-abl gene antisense oligonucleotides
Juan PING ; Na ZHAO ; Baoquan WANG ; Zhihui SHEN ; Mingxing YIN ; Xiaobin PANG ; Chuanbo CHEN
China Oncology 2015;(3):167-172
Background and purpose:As the development of the completion of the human genome project (HGP), the research focus is turning to the gene function research. At present, the domestic experimental research on the apoptosis of K562 cells induced by antisense olignonucleotides is rare. This study was aimed to investigate the effect of human chronic myelogenou leukemia (CML) bcr-abl fusion gene antisense oligonucletides on autophagy and apoptosis of CMLK562 cells in vitro. Methods:By liposome as the carrier, K562 cells were transfected with the bcr-abl gene antisense olignonucleotides. Hoechst staining method was used to observe the apoptosis inducing effect of different concentrations of oligonucleotides, the expressions of LC3-Ⅱ, autophagy-related protein, were determined by the Western blot method, the cell cycles were determined by lfow cytometry (FCM), and JEM-4000EX electron microscope technology was used to detect the apoptosis morphological changes. The apoptosis was detected by DNA agarose gel electrophoresis. Results:Hoechst staining results showed that the bcr-abl gene antisense oligonucletides signiifcantly promoted the apoptosis of K562 cells in a certain concentration dependent manner. Western blot showed that the expression level of LC3-Ⅱwas obviously higher in bcr-abl gene antisense oligonucletides transfected group than the control group, showing a promoting effect on cell autophagy. FCM test results showed that bcr-abl gene antisense oligonucleotides transfected K562 cells showed obvious cell cycle arrest, visible obvious apoptosis morphology under the electron microscope, and DNA Ladder showed obvious apoptosis fragments. Conclusion:The bcr-abl gene antisense olignonucleotides can signiifcantly induce the cell apoptosis of K562. This study provides a new method for CML therapy.
10.Correlation analysis between psychological counseling behaviors of accompanying family members and anxiety and depression of inpatients with hepatocellular carcinoma
Yuqun WEI ; Anhua LI ; Yingfan HUANG ; Xinhua ZHAO ; Juan TANG ; Zhangyang MO ; Yonghui PANG
Chinese Journal of Practical Nursing 2017;33(8):602-607
Objective To understand the correlation between psychological counseling behaviors of accompanying family members and anxiety and depression of inpatients with hepatocellular carcinoma. Methods Used self-designed questionnaires to investigate the behaviors of 45 accompanying family members psychological counseling for their relative hospitalized patients with hepatocellular carcinoma, at the same time, evaluated the anxiety and depression among those 45 hospitalized patients with hepatocellular carcinoma, then analyzed the correlation between the accompanying family members' psychological counseling behaviors and hepatocellular carcinoma in patients' anxiety and depression. Results Among the accompanying family members,53.3%(24/45) were the spouses of the patients. During the accompanying nursing time, there were 20.0%(9/45) accompanying families told to their relative patients less than 2 hours per day, and 80.0%(36/45) told to the patients over 2 hours per day. The proportion of anxiety and depression in the hospitalized patients with hepatocellular carcinoma was 28.9%(13/45) and 77.8% (34/45) respectively. Whether the relationship between the accompanying family members and the patients with hepatocellular carcinoma were spouse relationship was related to the incidence of depression in the patients (χ2=10.470, P=0.005), and whether the time accompanying family members spent in talking with the patients over 2 hours every day was correlative with the incidence of anxiety and depression in the patients (χ2=7.012, P =0.030 andχ2=6.438, P=0.040). The patients who accompanying families told to them less than 2 hours per day suffered a higher depression rate (8/9), and among the patients who accompanying families told to them over 2 hours per day, the lowest rate of anxiety (1/12) was found in the patients who accompanying families told to them around 4.1-6.0 hours per day. Conclusions The patients who are accompanied and cared by their spouses have lower depression rate and lesser depression extent. The accompanying families talk to the patients total 4.1-6.0 hours per day could most favorably reduce the incidence of anxiety and depression in the hospitalized patients with hepatocellular carcinoma.