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Lyophilized recombinant human brain natriuretic peptide: A promising therapy in patients with chronic heart failure

World Journal of Clinical Cases 2023年第36期11卷 8603-8605页

作者： Christos Kourek Alexandros Briasoulis Grigorios Giamouzis John Skoularigis Andrew Xanthopoulos Medical School of Athens National and Kapodistrian University of AthensAthens 15772Greece Department of Clinical Therapeutics Alexandra HospitalFaculty of MedicineNational and Kapodistrian University of AthensAthens 11528Greece Department of Cardiology University Hospital of LarissaLarissa 41110Greece

Lyophilized recombinant brain natriuretic peptide(BNP)is an exogenous peptide synthesized by artificial recombination technology,with a similar structure and similar physiological effects with the endogenous natriuretic peptide secreted by the human ***’s main mechanism of action is to increase cyclic guanosine monophosphate by binding with its corresponding receptor in the body,regulating,thus,the imbalance of the vascular system and cardiac hemodynamics,improving the heart’s pumping capacity,and inhibiting sympathetic excitability and myocardial ***,it can promote mitochondrial metabolism and enhance the use of adenosine triphosphate in *** the present study,102 chronic heart failure(HF)patients were randomly assigned to a control and an observation group consisting of 51 patients *** of the control group were treated with standard HF therapy for 3 d including oral metoprolol tartrate tablets,spironolactone,and olmesartanate while patients of the observation group were administered the recombinant human BNP injection for the same time-period,plus the standard HF *** recombinant human BNP group(observation group)demonstrated better physical,emotional,social,and economic scores,as well as cardiac and inflammatory biomarkers such as serum hypersensitive C-reactive protein,N-terminal pro BNP and troponin I levels,compared to the control ***,cardiac function was also improved,as left ventricular ejection fraction and stroke volume were significantly higher in the observation group than in the control ***,adverse reactions were not different between the 2 ***,these results are not generalizable and the need of large multicenter randomized controlled trials examining the safety and efficacy of recombinant human BNP in HF patients is of major importance.

关键词： Heart failure recombinant Brain natriuretic peptide Outcomes

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recombinant expressed vasoactive intestinal peptide analogue ameliorates TNBS-induced colitis in rats

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World Journal of Gastroenterology 2018年第6期24卷 706-715页

作者： Chun-lan Xu Yu Guo lei Qiao li Ma Yi-Yi Cheng The Key Laboratory for Space Bioscience and Biotechnology School of Life Sciences Northwestern Polytechnical University

AIM To investigate the modulatory effect of recombinantexpressed vasoactive intestinal peptide(VIP) analogue(rVIPa) on trinitrobenzene sulfonic acid(TNBS)-induced colitis in rats. METHODS Forty-eight rats were randomized into six groups: normal control group(Control), model control group(TNBS), ethanol treatment group(ETOH), and VIP treatment groups with different dosage(rVIPa_(1nmol), rVIPa_(2nmol), rVIPa_(4nmol)). Diarrhea and bloody stool were observed. Colonic damage was evaluated histologically. The levels of tumor necrosis factor-α( TNF-α), interleukin-10(Il-10), myeloperoxidase(MPO) and endotoxin in colonic tissue and serum were determined by enzyme-linked immunosorbent assay(ElISA). The expression of occludin, ZO-1, Toll-like receptor 4(TlR4),and nuclear factor-kappa B p65(NF-κBp65), IκBα, and p-IκBα were detected by Western blot. RESULTS Administration with 2 nmol rVIPa prevented TNBSinduced necrosis, hyperemia, swelling, inflammation, etc., pathologic changes observed in the inner surface of colon in experimental rats. Moreover, rVIPa significantly decreased colonic TNF-α level(P < 0.001), MPO activity(P < 0.001) and serum endotoxin level(P < 0.01), and remarkably increased colonic Il-10 content(P < 0.001) in rats with TNBS-induced colitis. Furthermore, compared to the TNBS-induced colitis group, 2 nmol rVIPa treatment up-regulated the levels of occludin(P < 0.05) and ZO-1(P < 0.05), NF-κB p65(P < 0.01) and IκBα(P < 0.001), and down-regulated the levels of TlR4. CONCLUSION rVIPa ameliorates TNBS-induced colonic injury and inflammation and effectively protected the intestinal mucosal barrier function in rats. The mechanism may be related to TlR4/NF-κB-mediated signaling pathway. rVIPa could be used as a new alternative therapy for intestinal inflammatory disorders.

关键词： Vasoactive intestinal peptide Intestinal mucosal barrier Tight junction Toll-like receptors recombinant expression

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Identification of Stable Quantitative Trait Loci for Sheath Blight Resistance Using recombinant Inbred Line

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Rice science 2019年第5期26卷 331-338页

作者： CHEN Yuan ZENG Yuxiang JI Zhijuan LIANG Yan WEN Zhihua YANG Changdeng State Key Laboratory of Rice Biology China National Rice Research Institute

To identify stable quantitative trait loci(QTLs)responsible for sheath blight resistance,a recombinant inbred line mapping population consisting of 219 lines was developed by crossing Lemont and Yangdao *** disease rating,average lesion length,maximum disease rating and maximum lesion length were assayed in six different environments.A total of 128 minor effect QTLs were detected by multiple interval *** QTLs explained less than 11.2%of the phenotypic variations individually,and 106 QTLs were clustered in 20 QTL-rich regions/putative *** QTL×environment interactions were detected at three putative loci(qSBR11.1,qSBR11.2 and qSBR11.3),indicating that these three loci were not *** other 17 stable loci(qSBR1.1,qSBR1.2,qSBR2.1,qSBR2.3,qSBR3.1,qSBR3.2,qSBR3.5,qSBR3.6,qSBR5.1,qSBR7.1,qSBR8.1,qSBR9.1,qSBR9.2,qSBR9.3,qSBR12.1,qSBR12.2 and qSBR12.4)provided a foundation for marker-assisted selection in *** of allelic effect on the 20 putative loci identified 7 highly stable loci,including qSBR3.2,qSBR7.1,qSBR8.1,qSBR9.2,qSBR9.3,qSBR12.1 and qSBR12.2.

关键词： rice sheath blight resistance quantitative trait locus recombinant inbred line

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PDI-, PPI- and chaperone-catalyzed refolding of recombinant human IL-2 and GM-CSF

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Science China Chemistry 1995年第4期38卷 429-437页

作者：徐明波孟文华马贤凯 Institute of Basic Medical Sciences Beijing 100850 China

The studies on PDI-, PP1- and chaperone-catalyzed refolding of recombinant human IL-2 and GM-CSF show that PDI can prevent the mismatch of disalfide bonds and formation of aggregates by interchains linkage, furthermore, PDI can correct, the mismatching of disulflde bonds in IL-2 isomers. PPI can increase the rate of folding reaction while chaperone can prevent the aggregation during the folding process. In addition, there is a synergistic effect between them.

关键词： recombinant protein folding PDI PPI chaperone.

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Immunization of Male Mice with a New recombinant GnRH Fusion Protein Reduces the Testicular Function

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Agricultural Sciences in China 2009年第3期8卷 380-385页

作者： FANG Fu-gui YANG Ya-ping LIU Ya ZHANG Yun-hai TAO Yong WANG Suo-lu PU Yong ZHANG Xiao-rong Department of Animal Medicine College of Animal Science and Technology Anhui Agricultural University Hefei 230036 P.R.China Department of Animal Science College of Animal Science and Technology Anhui Agricultural University Hefei 230036 P.R.China

The objective of the present study was to evaluate the effect of an immunogenic maltose-binding protein-gonadotropin releasing hormone （GnRH-6-MBP） using genetic engineering. The synthetic mammalian tandem repeated GnRH hexamer gene was inserted into the expression plasmid pMAL-c2x. recombinant GnRH-6-MBP protein was over- expressed in *** strain BL21. Amylose resin with affinity chromatograph was used to purify target protein. The reactiongenicity of fusion protein was identified by indirect enzyme linked immunosorbent assay （ELISA）, and the antigenicity and biological effects of GnRH-6-MBP were tested in mice. In the experiment, 20 male Kunming white mice of 20 d old were randomly divided into treatment and control group. Ten mice were immunized with 100 μg GnRH-6-MBP administered subcutaneously （s.c.） thrice at 2-week intervals with GnRH-6-MBP. Mice were sacrificed after 3 weeks following the booster injection, the testis was removed, weighed and measured, and the histological structure was observed. The reactiongenicity of fusion protein to GnRH antibody was much higher than the control. Active immunization against GnRH-6-MBP reduced remarkably （P 〈 0.01） the length and weight of the testis, and shortened the girth and width of the testis （P 〈 0.05）, and suppressed testicular spermatogenesis compared to the control mice. These results indicate that the recombinant GnRH-6-MBP acted as a strong immunogen and caused atrophy of the testis.

关键词： GnRH recombinant vaccine maltose-binding protein (MBP) immunocastration

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A novel recombinant,VPAC2-selective agonist enhancing insulin release and glucose disposal

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Acta Pharmacologica Sinica 2007年第4期28卷 526-533页

作者： Ngai-lik TAM Bio-engineering Institute of Ji-nan University

Aim:To obtain the recombinant,VPAC2-selective ( VPAC2:type 3 receptor ofpituitary adenylate cyclase activating polypeptide which shared by vasoactiveintestinal peptide) agonist with effects on glucose disposal by intein-mediated,single column ***:A gene encoding 32-amino acid peptidenamed rMBAY was designed and synthesized and cloned into Escherichia coliexpression vector,pKYB (NEB,USA).The recombinant vector was transferredinto E coli ER2566 strain and the target protein was overexpressed as a fusion tothe N-terminus of a self-cleavable affinity *** the fusion protein was puri-fled by chitin-affinity chromatography,the self-cleavage activity of the intein wasinduced by β-mercaptoethanol and the target peptide,rMBAY,was released fromthe chitin-bound intein ***:Approximately 53 mg rMBAY with the purityover 95% was obtained by single column purification from 1 L induced culturefermented in 5 L *** results of the competitive binding assay andcAMP accumulation assay indicated that the recombinant rMBAY had specialbinding selectivity and potency for *** recombinant peptide,rMBAY,enhanced insulin release and decreased the plasma glucose level after intraperito-neal injection (50 ng/kg) with a high concentration of glucose (1.8 mmol/kg) in theNIH ***:An efficient production procedure of a recombinant VPAC2-selective agonist with corresponding effects on glucose disposal was established.

关键词： intein recombinant VPAC2 agonist insulin

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Expression and purification of recombinant human serum albumin from selectively terminable transgenic rice

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2013年第10期14卷 867-874页

作者： Qing ZHANG Hui YU Feng-zhen ZHANG Zhi-cheng SHEN State Key Laboratory of Rice Biology and Institute of Insect Sciences Zhejiang University

Human serum albumin(HSA) is widely utilized for medical purposes and biochemical *** rice has proved to be an attractive bioreactor for mass production of recombinant HSA(rHSA).However,transgene spread is a major environmental and food safety concern for transgenic rice expressing proteins of medical *** study aimed to develop a selectively terminable transgenic rice line expressing HSA in rice seeds,and a simple process for recovery and purification of rHSA for economical *** HSA expression cassette was inserted into a T-DNA vector encoding an RNA interference(RNAi) cassette suppressing the CYP81A6 *** gene detoxifies the herbicide bentazon and is linked to the 5-enolpyruvylshikimate-3-phosphate synthase(EPSPS) cassette which confers glyphosate *** Sepharose Fast Flow(ANX FF) anion exchange chromatography coupled with Butyl Sepharose High Performance(Butyl HP) hydrophobic interaction chromatography was used to purify rHSA.A transgenic rice line,HSA-84,was obtained with stable expression of rHSA of up to 0.72% of the total dry weight of the dehusked rice *** line also demonstrated high sensitivity to bentazon,and thus could be killed selectively by a spray of bentazon.A two-step chromatography purification scheme was established to purify the rHSA from rice seeds to a purity of 99% with a recovery of 62.4%.Results from mass spectrometry and N-terminus sequencing suggested that the purified rHSA was identical to natural plasma-derived *** study provides an alternative strategy for large-scale production of HSA with a built-in transgene safety control mechanism.

关键词： recombinant human serum albumin (rHSA) Selectively terminable transgenic rice Purification

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Neuroprotective effects of recombinant human erythropoietin on facial motoneurons after facial nerve injury in rats

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Neural Regeneration Research 2008年第5期3卷 521-524页

作者： Yubing Dai Wenlong Luo Hongjiang Chen Jianghua Nie Li Fang Xiaofei Lai Jingjing Li Department of Otorhinolaryngology-Head ＆Neck Surgery. The Second Affiliated Hospital of Chongqing Medical University. Chongqing 400010 China Department of Otorhinolaryngology-Head ＆Neck Surgery The Third People＇s Hospital of Chongqing Chongqing 400014 China Department of Radiology People＇s Hospital of Changshou District Chongqing 401220 China

BACKGROUND： Erythropoietin and recombinant human erythropoietin （rhEPO） inhibit apoptosis of motor neurons caused by spinal cord injury and brain damage in rats. However, it still remains to be shown whether rhEPO can protect facial motoneurons （FMNs） as Well. OBJECTIVE： To test the neuroprotective effects of rhEPO on injured VMNs, as well as the influence on Caspase-3 expression. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiment. This study was performed at the Central Laboratory of Basic Medical College, Chongqing Medical University from January to October 2007. MATERIALS： Seventy-five female SD rats, weighing 210-230 g. rhEPO injection was provided by Sansheng pharmaceuticals company, Shenyang City, Liaoning Province, China, and the License number was HMLN S20010001. METHODS： A total of 75 female rats were randomly divided into rhEPO treatment, control, and sham operation groups, with 25 rats in each group. Rat models of facial nerve injury were established in the rhEPO treatment group and the control group by crushing the main trunk of the left facial nerve. Surgical microscopic observation of the facial nerve damage displayed perineurial disruption. The left stylomastoid foramen of the sham operation group were only exposed, but without nerve injury. The rhEPO treatment group was treated with rhEPO （5 000 U/kg, i.p.） once following injury and once a day for two weeks. The control and sham operation groups were treated with the same dose of normal saline （i.p.）, once following injury and once a day for two weeks. MAIN OUTCOME MEASURES： Rats were sacrificed 3, 7, 14, 21, and 28 days after injury, FMN survival after facial nerve injury was analyzed by Toluidine blue staining, and then survival ratios （L/R） were calculated. The number of apoptotic profiles in the injured FMNs were evaluated by TUNEL staining. Expression of Caspase-3 in the facial nucleus was detected by immunohistochemistry methods. RESULTS： A total of 75 rats were included in the

关键词： facial nerve motoneurons erythropoietin recombinant apoptosis Caspase

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Construction of HBV-specific ribozyme and its recombinant with HDV and their cleavage activity in vitro

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World Journal of Gastroenterology 2000年第3期6卷 377-380页

作者： Shu Juan Wen Kai Jun Xiang Zhen Hua Huang Rong Zhou Xue Zhong Qi Gene Center of Nanfang Hospital First Military Medical UniversityGuangzhou 510515Guangdong ProvinceChina School of Life Sciences University of Science and Technology of ChinaHefei 230027Anhui ProvinceChina

AIM To construct the recombinant of HDVcDNA and HBV-specific ribozyme gene byrecombinant PCR in order to use HDV as atransporting vector carrying HBV-specificribozyme into liver cells for inhibiting thereplication of *** We separately cloned the ribozyme（RZ）gene and recombinant DVRZ（comprisingHDV cDNA and HBV-specific ribozyme gene）intothe downstream of T7 promoter of pTAdv-Tvector and studied the in vitro cleavage activityof their transcripts（rRZ,rDVRZ）on target RNA（rBVCF）from in vitro transcription of HBV Cgene fragment（BVCF）.RESULTS Both the simple（rRZ）and therecombinant ribozyme rDVRZ could efficientlycatalyze the cleavage of target RNA（rBVCF）under different temperatures（37℃,42℃ and55℃）and Mg2+concentrations（10 mmol/L,15 mmol/L and 20 mmol/L）and their catalyticactivity tended to increase as the temperaturewas *** the activity of rRZ was evidentlyhigher than that of *** The recombinant of HDV cDNAand ribozyme gene had the potential of beingfurther explored and used in gene therapy of HBVinfection.

关键词： hepatitis B virus hepatitis D virus ribozyme gene recombinant DVRZ

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Comparison between recombinant human parathyroid hormone(1-34) and elcatonin in treatment of primary osteoporosis

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Asian Pacific Journal of Tropical Medicine 2015年第1期8卷 79-84页

作者： Yan Yang Xue-Jun Zhang Xian-Jun Zhu Lei Zhang Ming-Jing Bao Yang Xian Ji-Chuan Wu Li-Mei Liu Peng-Qiu Li Department of Endocrinology Sichuan Provincial People's Hospital

Objective:To evaluate the efficacy and safety of rhPTH(1-34) ***:Sixty palients with primary OP were randomly divided into two groups according to the ratio of 3:***(1-34) group(PTH group) was treated with subcutaneous injection of rhPTH(1-34) 20 μg daily for 18 months,and the elcalonin group(CT group) was treated with intramuscular injection of elcatonin 20 U weekly for 12 *** mineral density(BMD) of the lumbar spine 2-4(L_(2-4))and femoral neck,serum calcium and phosphorus,urinary calcium,serum hone specific alkaline phosphatase(BSAP).and urinary c-terminal telopeptides of type Ⅰ collagen/creatinine(uCTX-Ⅰ /Cn were tested at baseline,and *** 18 months after ***:In PTH *** of L_(2-4),at 6,*** 18 months,BDM of Femoral neck at 18 month,BSAP at 6 and 12 months and uCTX- Ⅰ /Cr at 6.12 and 18 months were all significantly *** CT *** of L_(2-4) at12 month and that of femoral neck at 12 and 18 months were significantly elevated,while HSAP was significantly decreased at 12 and 18 months,and no significant difference on CTX- Ⅰ /Cr was *** BMD growth and growth rate between two groups were *** group had better improvement in L_(2-4) BMD and growth rate than CT group at *** 18 *** growth and growth rale of femoral neck al 12 month and its growth at 18 month in CT group were higher than in PTH group,hut there was no significant difference between two groups regarding the growth rates at 18 ***,there were no significant differences regarding the rales ol adverse reactions between two ***:rhPTH(1—34),is safe and effective in the treatment of primary *** is superior to elcatonin in improving vertebral HMD at onset time,growth rate and growth range,but inferior to elcatonin at HMD of femoral neck.

关键词： recombinant human parathyroid hormone Bone density Primary osteoporosis Elcatonin

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