Mytilus contain abundant antimicrobial peptides(AMPs)that play a key role in the innate immunity.However,heterologous production of these AMPs remains challenging due to their short sequences,multiple disulfide bonds,...Mytilus contain abundant antimicrobial peptides(AMPs)that play a key role in the innate immunity.However,heterologous production of these AMPs remains challenging due to their short sequences,multiple disulfide bonds,and high content of cationic amino acids,which hinder functional expression in prokaryotic systems such as Escherichia coli.To establish a eukaryotic recombinant expression system for the AMPs of mussel and obtain recombinant mussel AMPs for subsequent studies,we reported the successful recombinant expression of myticofensin B1,a novel defensin-like AMP identified previously in Mytiluscoruscus,using the eukaryotic host Pichia pastoris.The codon-optimized gene encoding the mature myticofensin-B1(composed of 65 amino acid residues,including 6 conserved cysteine residues)was cloned into a pPICZαA vector and expressed in P.pastoris GS115.Structural fidelity of the recombinant peptide was confirmed by liquid chromatography-tandem mass spectrometry(LC-MS/MS),showing a molecular weight of 8849.9 Da,which was consistent with the theoretical prediction.Functional assays demonstrated a broad-spectrum antimicrobial activity of the recombinant myticofensin-B1,with stronger inhibition against Gram-negative bacteria.Scanning electron microscopy revealed different effects of the recombinant myticofensin-B1 against different bacteria.In addition,the recombinant myticofensin-B1 exhibited a very low hemolytic activity against sheep red blood cells and weak cytotoxicity against human A549 lung cancer cells.This study establishes P.pastoris as a powerful platform to produce functional mussel AMP and highlights the potential of the recombinant myticofensin-B1 as a therapeutic agent for aquaculture pathogens and infections.展开更多
The published article titled“Truncated Bid Overexpression Induced by Recombinant Adenovirus Cre/LoxP System Suppresses the Tumorigenic Potential of CD133+Ovarian Cancer Stem Cells”has been retracted from Oncology Re...The published article titled“Truncated Bid Overexpression Induced by Recombinant Adenovirus Cre/LoxP System Suppresses the Tumorigenic Potential of CD133+Ovarian Cancer Stem Cells”has been retracted from Oncology Research,Vol.25,No.4,2017,pp.595–603.展开更多
Recombinant tissue plasminogen activator is commonly used for hematoma evacuation in minimally invasive surgery following intracerebral hemorrhage.However,during minimally invasive surgery,recombinant tissue plasminog...Recombinant tissue plasminogen activator is commonly used for hematoma evacuation in minimally invasive surgery following intracerebral hemorrhage.However,during minimally invasive surgery,recombinant tissue plasminogen activator may come into contact with brain tissue.Therefore,a thorough assessment of its safety is required.In this study,we established a mouse model of intracerebral hemorrhage induced by type VII collagenase.We observed that the administration of recombinant tissue plasminogen activator without hematoma aspiration significantly improved the neurological function of mice with intracerebral hemorrhage,reduced pathological damage,and lowered the levels of apoptosis and autophagy in the tissue surrounding the hematoma.In an in vitro model of intracerebral hemorrhage using primary cortical neurons induced by hemin,the administration of recombinant tissue plasminogen activator suppressed neuronal apoptosis,autophagy,and endoplasmic reticulum stress.Transcriptome sequencing analysis revealed that recombinant tissue plasminogen activator upregulated the phosphoinositide 3-kinase/RAC-alpha serine/threonine-protein kinase/mammalian target of rapamycin pathway in neurons.Moreover,the phosphoinositide 3-kinase inhibitor LY294002 abrogated the neuroprotective effects of recombinant tissue plasminogen activator in inhibiting excessive apoptosis,autophagy,and endoplasmic reticulum stress.Furthermore,to specify the domain of recombinant tissue plasminogen activator responsible for its neuroprotective effects,various inhibitors were used to target distinct domains.It has been revealed that the epidermal growth factor receptor inhibitor AG-1478 reversed the effect of recombinant tissue plasminogen activator on the phosphoinositide 3-kinase/RAC-alpha serine/threonineprotein kinase/mammalian target of rapamycin pathway.These findings suggest that recombinant tissue plasminogen activator exerts a direct neuroprotective effect on neurons following intracerebral hemorrhage,possibly through activation of the phosphoinositide 3-kinase/RAC-alpha serine/threonine-protein kinase/mammalian target of rapamycin pathway.展开更多
Magnetic resonance imaging(MRI)is a powerful tool for diagnosing and monitoring brain diseases,but its low sensitivity can hinder early detection.To address this challenge,we utilized chemical exchange saturation tran...Magnetic resonance imaging(MRI)is a powerful tool for diagnosing and monitoring brain diseases,but its low sensitivity can hinder early detection.To address this challenge,we utilized chemical exchange saturation transfer(CEST)MRI,which greatly enhances sensitivity for detecting low-concentration compounds.In this study,we developed a CEST contrast agent based on a recombinant adeno-associated viruses(rAAVs)encoding the protamine-1(PRM1)MRI reporter gene.CEST MRI revealed that PRM1 contrast agent effectively highlighted caudate putamen region after injection of the rAAVs into the mouse brain,clearly distinguishing it from the surrounding tissue,with no observable damage.This method provides a sensitive,metal-free CEST contrast agent for in vivo brain cell detection,demonstrating potential for both diagnostic and therapeutic applications in brain diseases.展开更多
BACKGROUND Patients with acute-on-chronic liver failure(ACLF)have a high mortality rate,poor prognosis,and often experience concurrent thrombocytopenia and bleeding events.AIM To evaluate the efficacy and safety of re...BACKGROUND Patients with acute-on-chronic liver failure(ACLF)have a high mortality rate,poor prognosis,and often experience concurrent thrombocytopenia and bleeding events.AIM To evaluate the efficacy and safety of recombinant human thrombopoietin(rhTPO)in patients with ACLF with concomitant severe thrombocytopenia.METHODS This was a prospective,open-label study.We assigned 70 ACLF patients with severe thrombocytopenia into the rhTPO group and control group,with 35 patients in each group.Patients in the rhTPO group received subcutaneous injections of rhTPO at a dose of 15000 IU/day for 7 consecutive days,while patients in the control group did not receive rhTPO treatment.The primary endpoint was the proportion of patients with platelet count>50×10^(9)/L on day 14.RESULTS The proportion of patients with platelet count>50×10^(9)/L on day 14 was 60.7%in the rhTPO group,which was significantly higher than that(12.0%)in the control group(P<0.001).The platelet count in the rhTPO group on day 14 was 64×10^(9)/L,exceeding the baseline of 28×10^(9)/L.Compared to the control group,the rhTPO group exhibited a significant increase in platelet count from baseline(P<0.05).Model for end-stage liver disease score,albumin level and international normalized ratio improved significantly from baseline on day 14 after rhTPO injection.The concentrations of serum thrombopoietin and hepatocyte growth factor in the rhTPO group after 7 days were 143.7 and 195.4 pg/mL,respectively,showing a significant increase from baseline(P<0.05).Eight(22.9%)patients had bleeding events in the control group compared with four(11.4%)in the rhTPO group.The incidence of 90-day mortality was also higher in the control group(6,17.1%)than that in the rhTPO group(3,8.6%).CONCLUSION rhTPO significantly increased the platelet count in ACLF patients with thrombocytopenia and reduce the occurrence of bleeding events,with a good safety profile.展开更多
Postoperative cognitive dysfunction is a seve re complication of the central nervous system that occurs after anesthesia and surgery,and has received attention for its high incidence and effect on the quality of life ...Postoperative cognitive dysfunction is a seve re complication of the central nervous system that occurs after anesthesia and surgery,and has received attention for its high incidence and effect on the quality of life of patients.To date,there are no viable treatment options for postoperative cognitive dysfunction.The identification of postoperative cognitive dysfunction hub genes could provide new research directions and therapeutic targets for future research.To identify the signaling mechanisms contributing to postoperative cognitive dysfunction,we first conducted Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of the Gene Expression Omnibus GSE95426 dataset,which consists of mRNAs and long non-coding RNAs differentially expressed in mouse hippocampus3 days after tibial fracture.The dataset was enriched in genes associated with the biological process"regulation of immune cells,"of which Chill was identified as a hub gene.Therefore,we investigated the contribution of chitinase-3-like protein 1 protein expression changes to postoperative cognitive dysfunction in the mouse model of tibial fractu re surgery.Mice were intraperitoneally injected with vehicle or recombinant chitinase-3-like protein 124 hours post-surgery,and the injection groups were compared with untreated control mice for learning and memory capacities using the Y-maze and fear conditioning tests.In addition,protein expression levels of proinflammatory factors(interleukin-1βand inducible nitric oxide synthase),M2-type macrophage markers(CD206 and arginase-1),and cognition-related proteins(brain-derived neurotropic factor and phosphorylated NMDA receptor subunit NR2B)were measured in hippocampus by western blotting.Treatment with recombinant chitinase-3-like protein 1 prevented surgery-induced cognitive impairment,downregulated interleukin-1βand nducible nitric oxide synthase expression,and upregulated CD206,arginase-1,pNR2B,and brain-derived neurotropic factor expression compared with vehicle treatment.Intraperitoneal administration of the specific ERK inhibitor PD98059 diminished the effects of recombinant chitinase-3-like protein 1.Collectively,our findings suggest that recombinant chitinase-3-like protein 1 ameliorates surgery-induced cognitive decline by attenuating neuroinflammation via M2 microglial polarization in the hippocampus.Therefore,recombinant chitinase-3-like protein1 may have therapeutic potential fo r postoperative cognitive dysfunction.展开更多
[Objective]To design and express a recombinant protein rMKIBV incorporating confirmed antigenic epitopes of infectious bronchitis virus(IBV)as a vaccine to provide comprehensive protection.Additionally,it explores the...[Objective]To design and express a recombinant protein rMKIBV incorporating confirmed antigenic epitopes of infectious bronchitis virus(IBV)as a vaccine to provide comprehensive protection.Additionally,it explores the potential of polyclonal yolk antibodies(IgY)harvested from laying hens immunized with the rMKIBV vaccine in the prevention and control of IBV.[Methods]The antigenic epitope sequences of IBV,obtained from online databases,were compared with sequences of representative IBV strains from GenBank.Flexible peptides were designed to link all antigenic peptides.The constructed amino acid sequence was analyzed,reverse-translated,codon-optimized,and then inserted into the pET-28a(+)cloning vector.The recombinant vector was introduced into Escherichia coli for expression.The purified,desalted,and endotoxin-removed rMKIBV protein was used as a vaccine to immunize animals for investigation of its immunogenicity and ability to stimulate specific IgY production in laying hens.[Results]The retrieved IBV antigenic epitope sequences showed high similarity with the published N and S protein sequences of 22 representative IBV strains.The predicted isoelectric point and molecular weight of rMKIBV were 10.25 and 63.39 kDa,respectively.The secondary structure of rMKIBV included a high proportion of random coils,which suggested strong antigenicity.High-purity rMKIBV was obtained from E.coli transformed with the recombinant plasmid pET-28a-mkibv.This protein specifically bound to anti-His-tag antibodies,N protein antibodies,and S protein antibodies.The mice immunized with this protein showed increases in the spleen index(P<0.05),elevations in the levels of serum-specific IgG antibodies(P<0.01)and IFN-γ(P<0.05),and no significant change in the IL-2 level.Immunized laying hens successfully produced IgY in egg yolks,with specific IgY antibody levels significantly increasing.Moreover,the IgY antibody titer gradually rose after immunization,reaching the peak after about 50 days and then gradually declining to reach a stable level.[Conclusion]We successfully constructed and expressed the recombinant protein rMKIBV.The protein demonstrated good immunogenicity,stimulating specific antibody production in both mice and laying hens.Notably,the IgY extracted from the yolks of immunized laying hens offers a novel approach to IBV prevention and control.These findings hold significant scientific and practical value for the development of vaccines against IBV.展开更多
Recombinant human growth hormone(rhGH)has been widely used for the treatment of disorders associated with GH deficiency and multiple clinical indications[1].Accurate determination of biological activity is essential i...Recombinant human growth hormone(rhGH)has been widely used for the treatment of disorders associated with GH deficiency and multiple clinical indications[1].Accurate determination of biological activity is essential in the development,registration,and quality control of rhGH pharmaceutical products[2].However,the existing in vivo bioassay procedure based on somatropin-induced weight gain in rats is complicated,and the use of a rat cell line-based approach(Nb2-11 bioassay),which measures the production of adenosine triphosphate(ATP)as a direct indicator of cell growth,has a low mechanism of action(MOA)relevance.Therefore,novel rhGH bioassays are still needed.To this end,we developed a reporter gene assay(RGA)based on the GH/insulin-like growth factor-1(IGF-1)axis.展开更多
Dear editor,As of 2023,the domestic cat population in China reached 65 million,surpassing dogs to become the most numerous companion animal in the country.Feline calicivirus(FCV)infection,one of the three most prevale...Dear editor,As of 2023,the domestic cat population in China reached 65 million,surpassing dogs to become the most numerous companion animal in the country.Feline calicivirus(FCV)infection,one of the three most prevalent infectious diseases in cats,poses a severe threat to feline health.FCV,classified under the Caliciviridae family(genus Vesivirus).展开更多
The cohort study by Li et al provides timely and clinically relevant evidence on the use of recombinant human thrombopoietin(rhTPO)in pediatric allogeneic hematopoietic stem cell transplantation.The authors report enh...The cohort study by Li et al provides timely and clinically relevant evidence on the use of recombinant human thrombopoietin(rhTPO)in pediatric allogeneic hematopoietic stem cell transplantation.The authors report enhanced platelet engraftment and a favorable safety profile,particularly in younger children aged 0-9 years.This age-dependent difference not only highlights the physiological responsiveness of early hematopoietic environments to rhTPO but also raises important questions about tailoring supportive therapies across pediatric age groups.While the findings are promising,the lack of a control group and single-center limitations warrant further multicenter,long-term investigations.Ne-vertheless,the study lays a compelling foundation for integrating rhTPO more broadly into pediatric transplant protocols and for advancing individualized post-transplant care.展开更多
BACKGROUND The safety and efficacy of recombinant human thrombopoietin(rhTPO)administered after allogeneic hematopoietic stem cell transplantation(allo-HSCT)in children(0-9 years old)and adolescents(10-17 years old)wi...BACKGROUND The safety and efficacy of recombinant human thrombopoietin(rhTPO)administered after allogeneic hematopoietic stem cell transplantation(allo-HSCT)in children(0-9 years old)and adolescents(10-17 years old)with hematological disorders remain unclear.AIM To evaluate the safety and efficacy of rhTPO administered before platelet(PLT)engraftment in pediatric patients with hematological disorders undergoing HSCT,and to investigate its effects on the incidence of graft-vs-host disease(GVHD)and other transplant-related outcomes.METHODS This study enrolled 79 pediatric patients with hematological disorders who received rhTPO after allo-HSCT.The safety and tolerability of rhTPO were evaluated and compared in children(n=36)and adolescents(n=43)with hematological disorders.We also investigated the effects of rhTPO administration on the incidence of GVHD and other transplant-related outcomes.Additionally,we examined the efficacy of rhTPO after allo-HSCT in children and adolescents.RESULTS All of the children and adolescents underwent hematopoietic reconstruction.The median time to PLT engraftment was 16 days for all patients,with 14(range,11-24)days in the 0-to 9-year-old group and 16(range,11-41)days in the 10-to 17-year-old group;the difference was statistically significant(P<0.05).The median time to neutrophil engraftment was 12 days in both groups.The median recovery times for PLT counts of≥20×10^(9)/L and≥50×10^(9)/L in the 0-to 9-year-old group were 10(range,2-20)and 11(range,2-20)days,respectively,and those for the 10-to 17-year-old group were 9(range,4-23)and 12(range,5-34)days,respectively.Children exhibited significantly shorter time to PLT engraftment(14 days vs 16 days)and shorter recovery time to PLT count≥100×10^(9)/L(16 days vs 18 days)(P<0.05)than adolescents.The incidence of acute GVHD in all patients was 53.2%,with a higher incidence in children(61.1%)than in adolescents(46.5%).The incidence of chronic GVHD showed little difference between the two age groups,with an overall incidence of 10.1%.No adverse events,other than bleeding,were observed in either age group.The incidence of bleeding was 20.3%.The median follow-up time for all survivors was 573 days(range:42-1803 days)after transplantation.At the final follow-up,3 patients in the 0-to 9-year-old group died;however,none of these deaths were attributed to allo-HSCT or the use of rhTPO.All patients survived in the 10-to 17-year-old group.CONCLUSION rhTPO was not associated with any significant safety issues and was well tolerated by pediatric and adolescent patients with hematologic diseases who underwent allo-HSCT.Our results suggested that rhTPO may benefit allo-HSCT in children and adolescents by improving PLT recovery.展开更多
Objective Recombination events are common and serve as the primary driving force of diverse human adenovirus(HAdV),particularly in children with acute respiratory tract infections(ARIs).Therefore,continual monitoring ...Objective Recombination events are common and serve as the primary driving force of diverse human adenovirus(HAdV),particularly in children with acute respiratory tract infections(ARIs).Therefore,continual monitoring of these events is essential for effective viral surveillance and control.Methods Respiratory specimens were collected from children with ARIs between January 2022 and December 2023.The penton base,hexon,and fiber genes were amplified from HAdV-positive specimens and sequenced to determine the virus type.In cases with inconsistent typing results,genes were cloned into the pGEM-T vector to detect recombination events.Metagenomic next-generation sequencing(mNGS)was performed to characterize the recombinant HAdV genomes.Results Among 6,771 specimens,277(4.09%,277/6,771)were positvie for HAdV,of which 157(56.68%,157/277)were successfully typed,with HAdV-B3 being the dominant type(91.08%,143/157),and 14(5.05%,14/277)exhibited inconsistent typing results,six of which belonged to species B.The penton base genes of these six specimens were classified as HAdV-B7,whereas their hexon and fiber genes were classified as HAdV-B3,resulting in a recombinant genotype designated P7H3F3,which closely resembled HAdV-B114.Additionally,a partial gene encoding L152/55 kD was identified,which originated from HAdV-B16.Conclusion A novel recombinant,P7H3F3,was identified,containing sequences derived from HAdV-B3 and HAdV-B7,which is similar to HAdV-B114,along with additional sequences from HAdV-B16.展开更多
Objective:To investigate the clinical efficacy and safety evaluation of Polyethylene Glycol Recombinant Human Growth Hormone Injection(PEG-rhGH)in the treatment of idiopathic short stature.Methods:A total of 1402 pati...Objective:To investigate the clinical efficacy and safety evaluation of Polyethylene Glycol Recombinant Human Growth Hormone Injection(PEG-rhGH)in the treatment of idiopathic short stature.Methods:A total of 1402 patients were enrolled from March 21,2024 to January 13,2025,including 778 males and 624 females,with ages mainly ranging from 5 to 13 years old.Follow-up visits were completed by 488 patients for the first time,174 patients for the second time,and 81 patients for the third time.All patients were treated with PEG-rhGH(Jin Sai Zeng)as the main therapy after admission.The changes in height information,IGF-1,and thyroid examination results of each patient at the initial diagnosis,6,9,and 12 months after treatment were observed and analyzed.Results:There was no statistical difference between the baseline and the initial diagnosis,as well as the second follow-up visit(P<0.05),while there was a statistical difference between the baseline and the first and third follow-up visits(P>0.05).There was a statistically significant difference in IGF-1 between the initial diagnosis and the first follow-up visit(P<0.05),but no statistical difference between the first,second,and third follow-up visits(P>0.05).Additionally,IGF-1 levels increased with time.There was no statistical difference in TSH between the initial diagnosis and the first,second,and third follow-up visits(P>0.05).There was a statistical difference in free T3 between the initial diagnosis and the first and second follow-up visits(P<0.05),but no statistical difference between the second and third follow-up visits(P>0.05).There was no statistical difference in free T4 between the initial diagnosis and the first and second follow-up visits(P>0.05),but there was a statistical difference between the second and third follow-up visits(P<0.05).Conclusion:PEG-rhGH(Jin Sai Zeng)is significantly effective in improving height and IGF-1 levels in patients with idiopathic short stature.展开更多
目的探讨小剂量多巴胺联合重组人脑利钠肽(recombinant human brain natriuretic peptide,rhBNP)治疗心肌梗死(MI)后心力衰竭(HF)的临床效果及对患者系统性免疫炎症指数(SII)、三酰甘油葡萄糖乘积(TyG)指数的影响。方法选取64例MI后HF患...目的探讨小剂量多巴胺联合重组人脑利钠肽(recombinant human brain natriuretic peptide,rhBNP)治疗心肌梗死(MI)后心力衰竭(HF)的临床效果及对患者系统性免疫炎症指数(SII)、三酰甘油葡萄糖乘积(TyG)指数的影响。方法选取64例MI后HF患者,按随机数字表法分为对照组与观察组各32例,患者在入院后均给予常规治疗,对照组在常规治疗基础上给予rhBNP治疗,观察组在常规治疗基础上给予小剂量多巴胺联合rhBNP治疗。治疗7d后,观察2组患者血清SII、TyG指数与心功能指标[左室射血分数(LVEF)、前负荷率(EDFR)、外周血管阻力(SVR)]变化及临床效果、不良反应和主要不良心血管事件(MACE)发生情况。结果观察组患者治疗后SII、TyG水平低于对照组(P<0.05);心功能指标LVEF、SVR高于对照组,EDFR低于对照组(P<0.05)。观察组治疗总有效率(96.88%)高于对照组(84.38%)(P<0.05);治疗后6个月MACE发生率观察组(3.13%)低于对照组(18.75%)(P<0.05);用药不良反应发生率2组间比较差异无统计学意义(P>0.05)。结论采取小剂量多巴胺联合rhBNP用药方案治疗MI后HF,改善患者临床疗效、炎症状态及心功能较单一使用rhBNP效果更好,不明显增加不良事件发生率,值得基层医院参考应用。展开更多
基金supported by the National Natural Science Foundation of China(32271580,42020104009)the Fundamental Research Funds for Zhejiang Provincial Universities and Research Institutes(JX6311101923)。
文摘Mytilus contain abundant antimicrobial peptides(AMPs)that play a key role in the innate immunity.However,heterologous production of these AMPs remains challenging due to their short sequences,multiple disulfide bonds,and high content of cationic amino acids,which hinder functional expression in prokaryotic systems such as Escherichia coli.To establish a eukaryotic recombinant expression system for the AMPs of mussel and obtain recombinant mussel AMPs for subsequent studies,we reported the successful recombinant expression of myticofensin B1,a novel defensin-like AMP identified previously in Mytiluscoruscus,using the eukaryotic host Pichia pastoris.The codon-optimized gene encoding the mature myticofensin-B1(composed of 65 amino acid residues,including 6 conserved cysteine residues)was cloned into a pPICZαA vector and expressed in P.pastoris GS115.Structural fidelity of the recombinant peptide was confirmed by liquid chromatography-tandem mass spectrometry(LC-MS/MS),showing a molecular weight of 8849.9 Da,which was consistent with the theoretical prediction.Functional assays demonstrated a broad-spectrum antimicrobial activity of the recombinant myticofensin-B1,with stronger inhibition against Gram-negative bacteria.Scanning electron microscopy revealed different effects of the recombinant myticofensin-B1 against different bacteria.In addition,the recombinant myticofensin-B1 exhibited a very low hemolytic activity against sheep red blood cells and weak cytotoxicity against human A549 lung cancer cells.This study establishes P.pastoris as a powerful platform to produce functional mussel AMP and highlights the potential of the recombinant myticofensin-B1 as a therapeutic agent for aquaculture pathogens and infections.
文摘The published article titled“Truncated Bid Overexpression Induced by Recombinant Adenovirus Cre/LoxP System Suppresses the Tumorigenic Potential of CD133+Ovarian Cancer Stem Cells”has been retracted from Oncology Research,Vol.25,No.4,2017,pp.595–603.
基金supported by the National Natural Science Foundation of China,Nos.92148206,82071330(both to ZT)a grant from the Major Program of Hubei Province,No.2023BAA005(to ZT)+1 种基金a grant from the Key Research and Discovery Program of Hubei Province,No.2021BCA109(to ZT)the Research Foundation of Tongji Hospital,No.2022B37(to PZ)。
文摘Recombinant tissue plasminogen activator is commonly used for hematoma evacuation in minimally invasive surgery following intracerebral hemorrhage.However,during minimally invasive surgery,recombinant tissue plasminogen activator may come into contact with brain tissue.Therefore,a thorough assessment of its safety is required.In this study,we established a mouse model of intracerebral hemorrhage induced by type VII collagenase.We observed that the administration of recombinant tissue plasminogen activator without hematoma aspiration significantly improved the neurological function of mice with intracerebral hemorrhage,reduced pathological damage,and lowered the levels of apoptosis and autophagy in the tissue surrounding the hematoma.In an in vitro model of intracerebral hemorrhage using primary cortical neurons induced by hemin,the administration of recombinant tissue plasminogen activator suppressed neuronal apoptosis,autophagy,and endoplasmic reticulum stress.Transcriptome sequencing analysis revealed that recombinant tissue plasminogen activator upregulated the phosphoinositide 3-kinase/RAC-alpha serine/threonine-protein kinase/mammalian target of rapamycin pathway in neurons.Moreover,the phosphoinositide 3-kinase inhibitor LY294002 abrogated the neuroprotective effects of recombinant tissue plasminogen activator in inhibiting excessive apoptosis,autophagy,and endoplasmic reticulum stress.Furthermore,to specify the domain of recombinant tissue plasminogen activator responsible for its neuroprotective effects,various inhibitors were used to target distinct domains.It has been revealed that the epidermal growth factor receptor inhibitor AG-1478 reversed the effect of recombinant tissue plasminogen activator on the phosphoinositide 3-kinase/RAC-alpha serine/threonineprotein kinase/mammalian target of rapamycin pathway.These findings suggest that recombinant tissue plasminogen activator exerts a direct neuroprotective effect on neurons following intracerebral hemorrhage,possibly through activation of the phosphoinositide 3-kinase/RAC-alpha serine/threonine-protein kinase/mammalian target of rapamycin pathway.
基金financially supported by the National Natural Science Foundation of China(82127802,22374157)Strategic Priority Research Program,CAS(XDB0540000,XDC0170000)CAS Youth Interdisciplinary Team(JCTD-2022-13).In addition,Xin Zhou acknowledges the support from the Tencent Foundation through the XPLORER PRIZE.
文摘Magnetic resonance imaging(MRI)is a powerful tool for diagnosing and monitoring brain diseases,but its low sensitivity can hinder early detection.To address this challenge,we utilized chemical exchange saturation transfer(CEST)MRI,which greatly enhances sensitivity for detecting low-concentration compounds.In this study,we developed a CEST contrast agent based on a recombinant adeno-associated viruses(rAAVs)encoding the protamine-1(PRM1)MRI reporter gene.CEST MRI revealed that PRM1 contrast agent effectively highlighted caudate putamen region after injection of the rAAVs into the mouse brain,clearly distinguishing it from the surrounding tissue,with no observable damage.This method provides a sensitive,metal-free CEST contrast agent for in vivo brain cell detection,demonstrating potential for both diagnostic and therapeutic applications in brain diseases.
基金Supported by Tianjin Key Medical Discipline(Specialty)Construction Project,No.TJYXZDXK-034A.
文摘BACKGROUND Patients with acute-on-chronic liver failure(ACLF)have a high mortality rate,poor prognosis,and often experience concurrent thrombocytopenia and bleeding events.AIM To evaluate the efficacy and safety of recombinant human thrombopoietin(rhTPO)in patients with ACLF with concomitant severe thrombocytopenia.METHODS This was a prospective,open-label study.We assigned 70 ACLF patients with severe thrombocytopenia into the rhTPO group and control group,with 35 patients in each group.Patients in the rhTPO group received subcutaneous injections of rhTPO at a dose of 15000 IU/day for 7 consecutive days,while patients in the control group did not receive rhTPO treatment.The primary endpoint was the proportion of patients with platelet count>50×10^(9)/L on day 14.RESULTS The proportion of patients with platelet count>50×10^(9)/L on day 14 was 60.7%in the rhTPO group,which was significantly higher than that(12.0%)in the control group(P<0.001).The platelet count in the rhTPO group on day 14 was 64×10^(9)/L,exceeding the baseline of 28×10^(9)/L.Compared to the control group,the rhTPO group exhibited a significant increase in platelet count from baseline(P<0.05).Model for end-stage liver disease score,albumin level and international normalized ratio improved significantly from baseline on day 14 after rhTPO injection.The concentrations of serum thrombopoietin and hepatocyte growth factor in the rhTPO group after 7 days were 143.7 and 195.4 pg/mL,respectively,showing a significant increase from baseline(P<0.05).Eight(22.9%)patients had bleeding events in the control group compared with four(11.4%)in the rhTPO group.The incidence of 90-day mortality was also higher in the control group(6,17.1%)than that in the rhTPO group(3,8.6%).CONCLUSION rhTPO significantly increased the platelet count in ACLF patients with thrombocytopenia and reduce the occurrence of bleeding events,with a good safety profile.
基金supported by the National Natural Science Foundation of China,Nos.81730033,82171193(to XG)the Key Talent Project for Strengthening Health during the 13^(th)Five-Year Plan Period,No.ZDRCA2016069(to XG)+1 种基金the National Key R&D Program of China,No.2018YFC2001901(to XG)Jiangsu Provincial Medical Key Discipline,No.ZDXK202232(to XG)。
文摘Postoperative cognitive dysfunction is a seve re complication of the central nervous system that occurs after anesthesia and surgery,and has received attention for its high incidence and effect on the quality of life of patients.To date,there are no viable treatment options for postoperative cognitive dysfunction.The identification of postoperative cognitive dysfunction hub genes could provide new research directions and therapeutic targets for future research.To identify the signaling mechanisms contributing to postoperative cognitive dysfunction,we first conducted Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of the Gene Expression Omnibus GSE95426 dataset,which consists of mRNAs and long non-coding RNAs differentially expressed in mouse hippocampus3 days after tibial fracture.The dataset was enriched in genes associated with the biological process"regulation of immune cells,"of which Chill was identified as a hub gene.Therefore,we investigated the contribution of chitinase-3-like protein 1 protein expression changes to postoperative cognitive dysfunction in the mouse model of tibial fractu re surgery.Mice were intraperitoneally injected with vehicle or recombinant chitinase-3-like protein 124 hours post-surgery,and the injection groups were compared with untreated control mice for learning and memory capacities using the Y-maze and fear conditioning tests.In addition,protein expression levels of proinflammatory factors(interleukin-1βand inducible nitric oxide synthase),M2-type macrophage markers(CD206 and arginase-1),and cognition-related proteins(brain-derived neurotropic factor and phosphorylated NMDA receptor subunit NR2B)were measured in hippocampus by western blotting.Treatment with recombinant chitinase-3-like protein 1 prevented surgery-induced cognitive impairment,downregulated interleukin-1βand nducible nitric oxide synthase expression,and upregulated CD206,arginase-1,pNR2B,and brain-derived neurotropic factor expression compared with vehicle treatment.Intraperitoneal administration of the specific ERK inhibitor PD98059 diminished the effects of recombinant chitinase-3-like protein 1.Collectively,our findings suggest that recombinant chitinase-3-like protein 1 ameliorates surgery-induced cognitive decline by attenuating neuroinflammation via M2 microglial polarization in the hippocampus.Therefore,recombinant chitinase-3-like protein1 may have therapeutic potential fo r postoperative cognitive dysfunction.
文摘[Objective]To design and express a recombinant protein rMKIBV incorporating confirmed antigenic epitopes of infectious bronchitis virus(IBV)as a vaccine to provide comprehensive protection.Additionally,it explores the potential of polyclonal yolk antibodies(IgY)harvested from laying hens immunized with the rMKIBV vaccine in the prevention and control of IBV.[Methods]The antigenic epitope sequences of IBV,obtained from online databases,were compared with sequences of representative IBV strains from GenBank.Flexible peptides were designed to link all antigenic peptides.The constructed amino acid sequence was analyzed,reverse-translated,codon-optimized,and then inserted into the pET-28a(+)cloning vector.The recombinant vector was introduced into Escherichia coli for expression.The purified,desalted,and endotoxin-removed rMKIBV protein was used as a vaccine to immunize animals for investigation of its immunogenicity and ability to stimulate specific IgY production in laying hens.[Results]The retrieved IBV antigenic epitope sequences showed high similarity with the published N and S protein sequences of 22 representative IBV strains.The predicted isoelectric point and molecular weight of rMKIBV were 10.25 and 63.39 kDa,respectively.The secondary structure of rMKIBV included a high proportion of random coils,which suggested strong antigenicity.High-purity rMKIBV was obtained from E.coli transformed with the recombinant plasmid pET-28a-mkibv.This protein specifically bound to anti-His-tag antibodies,N protein antibodies,and S protein antibodies.The mice immunized with this protein showed increases in the spleen index(P<0.05),elevations in the levels of serum-specific IgG antibodies(P<0.01)and IFN-γ(P<0.05),and no significant change in the IL-2 level.Immunized laying hens successfully produced IgY in egg yolks,with specific IgY antibody levels significantly increasing.Moreover,the IgY antibody titer gradually rose after immunization,reaching the peak after about 50 days and then gradually declining to reach a stable level.[Conclusion]We successfully constructed and expressed the recombinant protein rMKIBV.The protein demonstrated good immunogenicity,stimulating specific antibody production in both mice and laying hens.Notably,the IgY extracted from the yolks of immunized laying hens offers a novel approach to IBV prevention and control.These findings hold significant scientific and practical value for the development of vaccines against IBV.
基金supported by the first batch of grants from the State Key Laboratory of Drug Regulatory Science,China(Grant No.:2023SKLDRS0108).
文摘Recombinant human growth hormone(rhGH)has been widely used for the treatment of disorders associated with GH deficiency and multiple clinical indications[1].Accurate determination of biological activity is essential in the development,registration,and quality control of rhGH pharmaceutical products[2].However,the existing in vivo bioassay procedure based on somatropin-induced weight gain in rats is complicated,and the use of a rat cell line-based approach(Nb2-11 bioassay),which measures the production of adenosine triphosphate(ATP)as a direct indicator of cell growth,has a low mechanism of action(MOA)relevance.Therefore,novel rhGH bioassays are still needed.To this end,we developed a reporter gene assay(RGA)based on the GH/insulin-like growth factor-1(IGF-1)axis.
基金supported by grants from the National Key Research and Development Program of China(Grant No.2022YFD1800100)National Natural Science Foundation of China(Grant No.32272982)Natural Science Foundation of Shanghai(Grant No.23ZR1477100).
文摘Dear editor,As of 2023,the domestic cat population in China reached 65 million,surpassing dogs to become the most numerous companion animal in the country.Feline calicivirus(FCV)infection,one of the three most prevalent infectious diseases in cats,poses a severe threat to feline health.FCV,classified under the Caliciviridae family(genus Vesivirus).
文摘The cohort study by Li et al provides timely and clinically relevant evidence on the use of recombinant human thrombopoietin(rhTPO)in pediatric allogeneic hematopoietic stem cell transplantation.The authors report enhanced platelet engraftment and a favorable safety profile,particularly in younger children aged 0-9 years.This age-dependent difference not only highlights the physiological responsiveness of early hematopoietic environments to rhTPO but also raises important questions about tailoring supportive therapies across pediatric age groups.While the findings are promising,the lack of a control group and single-center limitations warrant further multicenter,long-term investigations.Ne-vertheless,the study lays a compelling foundation for integrating rhTPO more broadly into pediatric transplant protocols and for advancing individualized post-transplant care.
文摘BACKGROUND The safety and efficacy of recombinant human thrombopoietin(rhTPO)administered after allogeneic hematopoietic stem cell transplantation(allo-HSCT)in children(0-9 years old)and adolescents(10-17 years old)with hematological disorders remain unclear.AIM To evaluate the safety and efficacy of rhTPO administered before platelet(PLT)engraftment in pediatric patients with hematological disorders undergoing HSCT,and to investigate its effects on the incidence of graft-vs-host disease(GVHD)and other transplant-related outcomes.METHODS This study enrolled 79 pediatric patients with hematological disorders who received rhTPO after allo-HSCT.The safety and tolerability of rhTPO were evaluated and compared in children(n=36)and adolescents(n=43)with hematological disorders.We also investigated the effects of rhTPO administration on the incidence of GVHD and other transplant-related outcomes.Additionally,we examined the efficacy of rhTPO after allo-HSCT in children and adolescents.RESULTS All of the children and adolescents underwent hematopoietic reconstruction.The median time to PLT engraftment was 16 days for all patients,with 14(range,11-24)days in the 0-to 9-year-old group and 16(range,11-41)days in the 10-to 17-year-old group;the difference was statistically significant(P<0.05).The median time to neutrophil engraftment was 12 days in both groups.The median recovery times for PLT counts of≥20×10^(9)/L and≥50×10^(9)/L in the 0-to 9-year-old group were 10(range,2-20)and 11(range,2-20)days,respectively,and those for the 10-to 17-year-old group were 9(range,4-23)and 12(range,5-34)days,respectively.Children exhibited significantly shorter time to PLT engraftment(14 days vs 16 days)and shorter recovery time to PLT count≥100×10^(9)/L(16 days vs 18 days)(P<0.05)than adolescents.The incidence of acute GVHD in all patients was 53.2%,with a higher incidence in children(61.1%)than in adolescents(46.5%).The incidence of chronic GVHD showed little difference between the two age groups,with an overall incidence of 10.1%.No adverse events,other than bleeding,were observed in either age group.The incidence of bleeding was 20.3%.The median follow-up time for all survivors was 573 days(range:42-1803 days)after transplantation.At the final follow-up,3 patients in the 0-to 9-year-old group died;however,none of these deaths were attributed to allo-HSCT or the use of rhTPO.All patients survived in the 10-to 17-year-old group.CONCLUSION rhTPO was not associated with any significant safety issues and was well tolerated by pediatric and adolescent patients with hematologic diseases who underwent allo-HSCT.Our results suggested that rhTPO may benefit allo-HSCT in children and adolescents by improving PLT recovery.
基金supported by the Capital’s Funds for Health Improvement and Research(CFH,shoufa-2022-1G-1131 and shoufa 2022-4G-1133)the High Level Technical Talent Construction Project of Beijing Municipal Health Commission(Discipline Leader-02-20)+1 种基金Beijing Municipal Public Welfare Development and Reform Pilot Project for Medical Research Institutes(JYY2023-10)the Pathogen Spectrum and Host Marker Analysis in Children with Respiratory Tract Infections of Children(Grant 2024-0040).
文摘Objective Recombination events are common and serve as the primary driving force of diverse human adenovirus(HAdV),particularly in children with acute respiratory tract infections(ARIs).Therefore,continual monitoring of these events is essential for effective viral surveillance and control.Methods Respiratory specimens were collected from children with ARIs between January 2022 and December 2023.The penton base,hexon,and fiber genes were amplified from HAdV-positive specimens and sequenced to determine the virus type.In cases with inconsistent typing results,genes were cloned into the pGEM-T vector to detect recombination events.Metagenomic next-generation sequencing(mNGS)was performed to characterize the recombinant HAdV genomes.Results Among 6,771 specimens,277(4.09%,277/6,771)were positvie for HAdV,of which 157(56.68%,157/277)were successfully typed,with HAdV-B3 being the dominant type(91.08%,143/157),and 14(5.05%,14/277)exhibited inconsistent typing results,six of which belonged to species B.The penton base genes of these six specimens were classified as HAdV-B7,whereas their hexon and fiber genes were classified as HAdV-B3,resulting in a recombinant genotype designated P7H3F3,which closely resembled HAdV-B114.Additionally,a partial gene encoding L152/55 kD was identified,which originated from HAdV-B16.Conclusion A novel recombinant,P7H3F3,was identified,containing sequences derived from HAdV-B3 and HAdV-B7,which is similar to HAdV-B114,along with additional sequences from HAdV-B16.
文摘Objective:To investigate the clinical efficacy and safety evaluation of Polyethylene Glycol Recombinant Human Growth Hormone Injection(PEG-rhGH)in the treatment of idiopathic short stature.Methods:A total of 1402 patients were enrolled from March 21,2024 to January 13,2025,including 778 males and 624 females,with ages mainly ranging from 5 to 13 years old.Follow-up visits were completed by 488 patients for the first time,174 patients for the second time,and 81 patients for the third time.All patients were treated with PEG-rhGH(Jin Sai Zeng)as the main therapy after admission.The changes in height information,IGF-1,and thyroid examination results of each patient at the initial diagnosis,6,9,and 12 months after treatment were observed and analyzed.Results:There was no statistical difference between the baseline and the initial diagnosis,as well as the second follow-up visit(P<0.05),while there was a statistical difference between the baseline and the first and third follow-up visits(P>0.05).There was a statistically significant difference in IGF-1 between the initial diagnosis and the first follow-up visit(P<0.05),but no statistical difference between the first,second,and third follow-up visits(P>0.05).Additionally,IGF-1 levels increased with time.There was no statistical difference in TSH between the initial diagnosis and the first,second,and third follow-up visits(P>0.05).There was a statistical difference in free T3 between the initial diagnosis and the first and second follow-up visits(P<0.05),but no statistical difference between the second and third follow-up visits(P>0.05).There was no statistical difference in free T4 between the initial diagnosis and the first and second follow-up visits(P>0.05),but there was a statistical difference between the second and third follow-up visits(P<0.05).Conclusion:PEG-rhGH(Jin Sai Zeng)is significantly effective in improving height and IGF-1 levels in patients with idiopathic short stature.
文摘目的探讨小剂量多巴胺联合重组人脑利钠肽(recombinant human brain natriuretic peptide,rhBNP)治疗心肌梗死(MI)后心力衰竭(HF)的临床效果及对患者系统性免疫炎症指数(SII)、三酰甘油葡萄糖乘积(TyG)指数的影响。方法选取64例MI后HF患者,按随机数字表法分为对照组与观察组各32例,患者在入院后均给予常规治疗,对照组在常规治疗基础上给予rhBNP治疗,观察组在常规治疗基础上给予小剂量多巴胺联合rhBNP治疗。治疗7d后,观察2组患者血清SII、TyG指数与心功能指标[左室射血分数(LVEF)、前负荷率(EDFR)、外周血管阻力(SVR)]变化及临床效果、不良反应和主要不良心血管事件(MACE)发生情况。结果观察组患者治疗后SII、TyG水平低于对照组(P<0.05);心功能指标LVEF、SVR高于对照组,EDFR低于对照组(P<0.05)。观察组治疗总有效率(96.88%)高于对照组(84.38%)(P<0.05);治疗后6个月MACE发生率观察组(3.13%)低于对照组(18.75%)(P<0.05);用药不良反应发生率2组间比较差异无统计学意义(P>0.05)。结论采取小剂量多巴胺联合rhBNP用药方案治疗MI后HF,改善患者临床疗效、炎症状态及心功能较单一使用rhBNP效果更好,不明显增加不良事件发生率,值得基层医院参考应用。
