1.Modification,Expression and Purification of Human Endotoxin Binding Peptide Gene
Ya-Li SUN ; You-Sheng LIU ; Hai-Jie YANG ;
China Biotechnology 2006;0(03):-
Objective: To modify EBP(endotoxin binding peptide), clone and express the mutate of EBP gene and gain purified mEBP.Method: mEBPgene was cloned by PCR site-directed mutagenesis. PinpointXa-3/mEBP expression vector was designed to express human mEBP as a fusion protein in BL21 (DE3) pLysS. Digested engineering bacteria by lysozyme and collected inclusion bodies.Fusion protein was purified by Pinpoint TM Xa purification system and cleaved by factorXa,mEBP was purified by RP-HPLC. Results: Mutations at residues 5 and 18(Gln→Lys) was obtained by PCR site-directed mutagenesis, expressed and purified mEBP successfully.Conclusions: Obtaining of purified mEBP lay a foundation for its biological activity research.
2.Effect of Yixintai Granule on mRNA and protein expression levels of AQP, in renal medulla of chronic heart failure rabbits.
Yun TANG ; Zhi-Hua GUO ; Ya LI ; Tao SUN ; Gang-Qiang WU ; Li LIU ; Jie DENG
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(3):333-337
OBJECTIVETo explore the effect of Yixintai Granule (YG) on mRNA and protein expression levels of AQP2 in renal medulla of chronic heart failure (CHF) rabbits.
METHODSCHF rat model was established by ear marginal vein injection of adriamycin. Successfully modeled rabbits were divided into the model group, the high (8.4 g/kg), middle (4.2 g/kg), and low dose (2.1 g/kg) YG group, and the Furosemide group (2 mg/kg). Besides, a normal control group was set up. Equal volume of physiological saline was administered to rabbits of the model group and the normal control group by gastrogavage. YG at different doses was administered to rabbits of the 3 YG groups by gastrogavage. The intervention lasted for 4 weeks, once per day. After treatment the urine volume and pathomorphological changes of renal medulla tissue were observed. mRNA and its protein expression levels of AQP2 were detected.
RESULTSCompared with the normal control group, the urine volume decreased significantly, mRNA and protein expression levels of renal medulla AQP2 increased significantly in the model group (all P < 0.01). Compared with the model group, the urine volume increased significantly, and mRNA and protein expression levels of renal medulla AQP2 decreased significantly in all medicated groups (all P < 0.01). Compared with the low dose YG group, the urine volume significantly increased and the mRNA expression level of renal medulla AQP2 significantly decreased in the middle and high dose YG groups (all P < 0.01). The expression level of AQP2 protein significantly decreased in the high dose YG group (P < 0.01). Pathological changes of the renal medulla was the most obviously seen in the model group. But they were alleviated to various degrees in all medicated groups. They were more obviously attenuated in the middle and high dose YG groups.
CONCLUSIONYG could improve CHF possibly through down-regulating mRNA and protein expression levels of AQP2 in renal medulla, and elevating the urine volume.
Animals ; Aquaporin 2 ; genetics ; metabolism ; Chronic Disease ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Heart Failure ; drug therapy ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Rats, Sprague-Dawley
3.Analysis of compound heterozygous mutation of low-density lipoprotein receptor gene in two patients with familial hypercholesterolemia
Yanfang DAI ; Xiaodong PAN ; Liyuan SUN ; Ya YANG ; Li SONG ; Jie LIN ; Luya WANG
Chinese Journal of Laboratory Medicine 2011;34(5):454-458
Objective To determine LDLR gene mutation in 2 clinically diagnosed FH patients from Hubei province and provide basis for gene diagnosis of FH.Methods Clinical data of 2 FH patients and their parents were collected.The promoter region and exon 1 to exon 18 region of LDLR gene were amplified through PCR and the amplified products were analyzed by forward and reverse DNA sequencing.The mutations were identified after comparison with LDLR gene sequence in GenBank.The pathogenic gene mutations were confirmed according to both genotype and phenotype of FH probands.Results The levels of plasma TC of two probands were 12.79 and 11.98 mmol/L.respectively.No gene mutations were detected in region 3 500 to 3 531 of ApoB100. The mutations of LDLR gene were compound heterozygous mutations. The novel mutation 665G > T detected in the exon 4 of No. 1 proband's LDLR gene was heterozygous missense mutation. The novel mutation 1 358 +32C > T was detected in the exon 9 of No. 1 proband's LDLR gene.The mutations 665G > T ( paternal origin) and 1 358 + 32C > T ( maternal origin) were inherited from the parents. A novel mutation 1 257 C > A was detected in the exon 9 of No. 2 proband's LDLR gene, resulting the presence of a premature termination codon, which was different from 1 257 C > G reported in Belgium.Another heterozygous missense mutation 1 879 G > A was detected in exon 13. They were derived from paternal origin and maternal origin, respectively. Conclusions There are three novel gene mutations:665G >T, 1 358 +32C > T, 1 257C > A found in two probands with compound heterozygous mutations in LDLR respectively. They maybe play a potential role in FH pathogensis.
4.Ultrasound evaluation of children with recurrent intussusception and relationship with enlarged mesenteric lymph nodes
Jing, PENG ; Ya-ping, ZHAO ; Qing, ZHANG ; Zhao-qun, HE ; Jie, SUN
Chinese Journal of Medical Ultrasound (Electronic Edition) 2010;07(12):2151-2155
Objective To evaluate the relationship between recurrent intussusception and enlarged mesenteric lymph nodes (EMLNs) in children by ultrasound.Methods A total of 35 cases (a total of 75 relapse) with imaging features of recurrent intussusception were retrospectively analysed.The sequence,size,number,morphology and location of EMLNs were recorded.Results Twenty cases of EMLNs were found in 35 cases recurrent intussusception (57.1%,20/35).At least three EMLNs were found in and / or around the mesentery of intussusception.The imaging feature of recurrent intussusception was a " target" appearance on transverse scans and a "sleeve" sign on longitudinal scans .All the EMLNs were smooth and oval-shaped hypoechoic nodule.The largest longitudinal diameter of the EMLNs ranged between 7-20 mm ,the largest transverse diameter of the EMLNs ranged between 3-8 mm with the aspect ratio≥2.0.Conclusion The Enlarged mesenteric lymph node recurrence of intussusception in children is crucial factor.Ultrasound can not only accurately and timely diagnose and treat intussusception while scanning enlarged mesenteric lymph nodes,which can provide an important basis for reducing the recurrence of intussusception.
5.Immunoreactivity Studies of Various HEV Antigen with Anti-HEV IgM of the Sear from Patients Infected with Hepatitis E Virus
Guang-ya, RONG ; Ji-wen, ZHOU ; Li, LEI ; Gui-lan, ZHAO ; Jie, SUN
Virologica Sinica 2001;16(2):128-130
Objective To compare the Immunoreactivity of various HEV Antigen with Anti-HEV IgM. Methods Solid-phase enzyme immunoassay( EIA ) was developed for detecting anti-HEV IgM by using synthetic peptides E30, E42, E33, and recombinant antigen from HEV ORF-2. Results Of 60 anti-HEV positive sera by using E30, E42, E33 and recombinant antigen as coating antigen, Anti-HEV IgM positive rates were 76.7%, 26.6%, 18.3% and 66.7% respectively. In Acute-phase and convalescence-phase sera of the patients with Hepatitis E, Anti-HEV IgM positive rate was 90% and 3.3% respectively. Conclusions The HEV E30-based EIA will be very useful in the early diagnosis of Hepatitis E.
6.The analysis of Keshan disease surveillance results in Yongjin Village, Fuyu County, Heilongjiang Province in 2007
Li-jun, ZHANG ; Hui, SUN ; Bai-nan, XU ; Jing, DENG ; Jie, HOU ; Zi-dan, GUO ; Ya-fei, SUN ; Xiao-ya, WANG ; Tong, WANG
Chinese Journal of Endemiology 2008;27(4):425-427
Objective To investigate the prevalence and incidence of Keshan disease (KD) and the selenium concentration of food and hair in residents of Yongjin Village, Fuyu County, Heilongjiang Province, national monitoring site, in 2007. Methods According to the Standard of Keshan Disease Surveillance and the Standard of Diagnosis of Keshan Disease(GB 17021-1997), the residents living in the monitoring site were surveyed by clinical examination and electrocardiography. For individuals whose hearts showed abnormalities, a chest X-ray photograph was taken. The selenium concentrations of the residents' food (flour) and hair were assayed by flowing injection hydride generation atomic fluoremetric method(FI-HG-AFM). Results Nineteen KD patients were found from 282 residents in 2007 KD surveillance. The prevalence of KD, latent KD and chronic KD were 6.7%(19/282), 2.8%(8/282) and 3.9%(11/282), respectively. Five of the 8 latent KD cases were newly found. In addition, there were 5 the suspected KD cases, including 2 suspected chronic KD cases. No acute KD or sub-acute KD patients were found in Yongjin Village at this monitoring site this year. The average selenium concentration of children hair and residents food were (0.3197±0.0586)mg/kg and (0.0210±0.0062)mg/kg, respectively. Conclusions New cases of KD continued to emerge, indicating that etiological factors still exist. Therefore, the emphasis of monitoring KD in furore is founding the consummate report of infectious disease system and training the personnel to increase the reliability of monitoring.
7.Analysis of acupoint-selection patterns in acupuncture-moxibustion treatment of polycystic ovarian syndrome based on data mining
Tao LI ; Jing ZHOU ; Huan-Gan WU ; Jie SUN ; Ya-Jing GUO ; Yi SUN ; Yin SHI ; Liang LI
Journal of Acupuncture and Tuina Science 2018;16(6):416-424
Objective:To analyze the meridian and acupoint-selection patterns in acupuncture-moxibustion treatment of polycystic ovarian syndrome (PCOS) by data mining technique,for exploring acupoints that could be effective for PCOS and providing suggestion and reference in selecting acupoints for the acupuncture-moxibustion treatment of PCOS.Methods:Literatures related to the acupuncture-moxibustion treatment of PCOS published before April 2018 were collected.By a self-made data mining program developed using Microsoft Excel 2007,a database of acupuncture-moxibustion treatment of PCOS was then established.The correlation analysis was performed for the frequency of use of acupoints and acupoint groups.Cluster analysis was also conducted.Results:A total of 92 articles were recruited finally.Sanyinjiao (SP 6),Conception Vessel and chest-abdomen region ranked the top on the list of the most frequently used acupoints,meridians and regions,respectively.Correlation analysis showed that the acupoint group with the highest confidence was Sanyinjiao (SP 6) and Zusanli (ST 36),and the group with the highest support was Sanyinjiao (SP 6) and Guanyuan (CV 4).Cluster analysis revealed five effective clusters:Guanyuan (CV 4) and Sanyinjiao (SP 6);Zhongwan (CV 12) and Tianshu (ST 25);Zhongji (CV 3),Zusanli (ST 36) and Qihai (CV 6);Xuehai (SP 10),Shenshu (BL 23),Taichong (LR 3) and Guilai (ST 29);Ganshu (BL 18),Taixi (KI 3),Yinlingquan (SP 9) and Pishu (BL 20).Conclusion:According to the confidence and support report,Sanyinjiao (SP 6) and Zusanli (ST 36) or Sanyinjiao (SP 6) and Guanyuan (CV 4) are recommended in acupuncture-moxibustion treatment of PCOS,with adjunct points selected for different patterns,e.g.Zhongwan (CV 12) and Tianshu (ST 25) for tonifying spleen and expelling dampness;Zhongji (CV 3) and Qihai (CV 6) for supplementing qi and cultivating blood;Xuehai (SP 10),Shenshu (BL 23),Taichong (LR 3) and Guilai (ST 29) for tonifying kidney and dissolving stasis;Ganshu (BL 18),Taixi (KI 3),Yinlingquan (SP 9) and Pishu (BL 20) for tonifying kidney and spleen and regulating liver.
8.Preparation of polyelectrolyte microcapsules contained gold nanoparticles.
Ya-jie SUN ; Jia-bi ZHU ; Chun-li ZHENG
Acta Pharmaceutica Sinica 2010;45(3):371-375
In this work, polyelectrolyte microcapsules containing gold nanoparticles were prepared via layer by layer assembly. Gold nanoparticles and poly (allyamine hydrochloride) (PAH) were coated on the CaCO3 microparticles. And then EDTA was used to remove the CaCO3 core. Scanning electron microscopy (SEM) was used to characterize the surface of microcapsules. SEM images indicate that the microcapsules and the polyelectrolyte multilayer were deposited on the surface of CaCO3 microparticles. FITC-bovine serum albumin (FITC-BSA, 2 mg) was incorporated in the CaCO3 microparticles by co-precipitation. Fluorescence microscopy was used to observe the fluorescence intensity of microcapsules. The encapsulation efficiency was (34.31 +/- 2.44) %. The drug loading was (43.75 +/- 3.12) mg g(-1).
Calcium Carbonate
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chemistry
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Capsules
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Drug Carriers
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Drug Delivery Systems
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methods
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Electrolytes
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chemistry
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Fluorescein-5-isothiocyanate
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analogs & derivatives
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chemistry
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Gold
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chemistry
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Microscopy, Electron, Scanning
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Microscopy, Fluorescence
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Nanoparticles
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Particle Size
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Serum Albumin, Bovine
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chemistry
9.An epidemiological study for no-suicide self-injury behavior of 1463 junior school students in Dalian city
Lei WANG ; Yueji SUN ; Yuan LIN ; Xin JIN ; Jie LIANG ; Guoqing XU ; Ya ZHENG ; Huijuan SHEN ; Chengqing ZHU
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(2):177-180
ObjectiveTo find incidence rate of no-suicide self injury (NSSI) of junior school students in Dalian city,and to provide some evidences for interventions for them.MethodsRandomly drawn out 1463 junior school students were served as study objects.All objects were evaluated with Ottawa Self-Injury Inventory (OSI) and self-made investigate questionnaire,75 no-suicide self-injury were screening out.ResultsThe incidence ratio of NSSI of junior school students was 5.4%,there were no significant different between the male and female students.The highest incidence ratio was found at 13 years old.The average age for first-time self-harm was (12.24±1.344) years old.There were no significant different between the male and the female students( t=- 1.415,P =0.163 ) ; Cutting Skin was the most common way of NSSI ( 12.0% ) ; Ideation of 80% of the NSSI was from their Own Idea.2.7% of the NSSI can Feel Relief through NSSI behavior.93% of NSSI was to regulate their mood.100% students of NSSI against NSSI behavior byReading Books or Listening Music,in which 60% of NSSI believe the method was helpful to relax their mind.78.5% of the NSSI resisted NSSI behavior by Watching TV or Playing Games,but they did not get enough effects.60% NSSI considered themselves without the need to treat.41.3% of NSSI had never been to treat.2.5% of NSSI went to hospital for the wound.ConclusionNSSI is often be found in junior school students,and highest ratio is at 13 years old.The most common method of NSSI is Cutting Skin.More NSSI aim is to release their emotion,and self-injury behavior accordance with their inner thoughts.NSSI behavior often is secret,and reading and listening to music is cut off from the relative effective way to conduct.NSSI are seldom to initiative doctor,and education organ,parents and society in a three-dimensional one of the system is necessary.