Dear Editor,Understanding the complex interactions between transcription factors(TFs)and DNA is crucial in various scientific fields,including genetics,molecular biology,and biochemistry.Current methodologies such as ...Dear Editor,Understanding the complex interactions between transcription factors(TFs)and DNA is crucial in various scientific fields,including genetics,molecular biology,and biochemistry.Current methodologies such as yeast one-hybrid,chromatin immunoprecipitation followed by sequencing,the firefly luciferase(Luc)reporter system,and DNA electrophoretic mobility shift assays often necessitate complicated procedures and costly chemicals(Ferraz et al.,2021).The adoption of the pigmentation-based reporter RUBY has facilitated the study of various plant signaling processes due to the easily observable nature of pigments(He et al.,2020).However,pigmentation-based assays lack sensitivity,and their high background limits broader applications.The recent discovery of the fungal Luc coding gene Luz in the bioluminescence pathway from the fungus Neonothopanus nambi offers new opportunities for enhancing reporter systems in transcriptional regulation assays(Kotlobay et al.,2018).The fungal bioluminescence pathway(FBP)is a cyclic metabolic route that involves the action of four enzymes:HispS,H3H,Luz,and CPH(Figure 1A).The pathway’s initial committed step utilizes caffeic acid as a substrate by the fungus HispS;NPGA from Aspergillus nidulans is capable of post-translationally modifying HispS.This modification enhances the activity of HispS,thereby increasing the intensity of FBP(Khakhar et al.,2020;Zheng et al.,2023;Shakhova et al.,2024).Notably,the synthesis pathway of caffeic acid has evolutionarily conserved homologs in plants(Supplemental Figure 1),suggesting that plant cells might independently utilize their own metabolic pool of caffeic acid to support the FBP without the need for additional substrate.This finding underscores the potential of FBP as a promising reporter for plant transcriptional regulation without substrate addition.展开更多
CRISPR-Cas-based genome editing holds immense promise for advancing plant genomics and crop enhancement.However,the challenge of low editing activity complicates the identification of editing events.In this study,we i...CRISPR-Cas-based genome editing holds immense promise for advancing plant genomics and crop enhancement.However,the challenge of low editing activity complicates the identification of editing events.In this study,we introduce multiple single transcript unit surrogate reporter(STU-SR)systems to enhance the selection of genome-edited plants.These systems use the same single guide RNAs designed for endogenous genes to edit reporter genes,establishing a direct link between reporter gene editing activity and that of endogenous genes.Various strategies are used to restore functional reporter genes after genome editing,including efficient single-strand annealing(SSA)for homologous recombination in STUSR-SSA systems.STU-SR-base editor systems leverage base editing to reinstate the start codon,enriching C-to-T and A-to-G base editing events.Our results showcase the effectiveness of these STU-SR systems in enhancing genome editing events in the monocot rice,encompassing Cas9 nuclease-based targeted mutagenesis,cytosine base editing,and adenine base editing.The systems exhibit compatibility with Cas9 variants,such as the PAM-less SpRY,and are shown to boost genome editing in Brassica oleracea,a dicot vegetable crop.In summary,we have developed highly efficient and versatile STU-SR systems for enrichment of genome-edited plants.展开更多
BACKGROUND Thrombophilia contributes to a significant increased risk of venous thromboembolism and can be either inherited or acquired.Hereditary thrombophilia may arise from various gene mutations,some of which have ...BACKGROUND Thrombophilia contributes to a significant increased risk of venous thromboembolism and can be either inherited or acquired.Hereditary thrombophilia may arise from various gene mutations,some of which have not even been adequately reported or poorly understood.Previous studies reported a rare and novel missense mutation in the prothrombin gene(p.Arg596Gln),known as prothrombin Belgrade.The mechanisms and therapeutic strategies associated with prothrombin Belgrade mutation have not been fully elucidated.CASE SUMMARY We present the case of a 26-year-old woman with recurrent systemic thrombosis induced by prothrombin Belgrade mutation.The patient suffered from cerebral venous sinus thrombosis that rapidly progressed to systemic thrombosis,alongside a family history of cerebral thrombosis,and no traditional risk factors or abnormal coagulation function.Whole-genome sequencing detected a novel and rare heterozygous prothrombin missense mutation,c.1787G>T(p.Arg596Gln),which was responsible for the major etiology of the systemic thrombosis.CONCLUSION This case strengthens our understanding about hereditary basis of thrombophilia and provokes considerations for therapeutic options on prothrombin Belgrade mutation.展开更多
BACKGROUND Meniscal tears are one of the most common knee injuries.After the diagnosis of a meniscal tear has been made,there are several factors physicians use to guide clinical decision-making.The influence of time ...BACKGROUND Meniscal tears are one of the most common knee injuries.After the diagnosis of a meniscal tear has been made,there are several factors physicians use to guide clinical decision-making.The influence of time between injury and isolated meniscus repair on patient outcomes is not well described.Assessing this relationship is important as it may influence clinical decision-making and can add to the preoperative patient education process.We hypothesized that increasing the time from injury to meniscus surgery would worsen postoperative outcomes.AIM To investigate the current literature for data on the relationship between time between meniscus injury and repair on patient outcomes.METHODS PubMed,Academic Search Complete,MEDLINE,CINAHL,and SPORTDiscus were searched for studies published between January 1,1995 and July 13,2023 on isolated meniscus repair.Exclusion criteria included concomitant ligament surgery,incomplete outcomes or time to surgery data,and meniscectomies.Patient demographics,time to injury,and postoperative outcomes from each study were abstracted and analyzed.RESULTS Five studies met all inclusion and exclusion criteria.There were 204(121 male,83 female)patients included.Three of five(60%)studies determined that time between injury and surgery was not statistically significant for postoperative Lysholm scores(P=0.62),Tegner scores(P=0.46),failure rate(P=0.45,P=0.86),and International Knee Documentation Committee scores(P=0.65).Two of five(40%)studies found a statistically significant increase in Lysholm scores with shorter time to surgery(P=0.03)and a statistically significant association between progression of medial meniscus extrusion ratio(P=0.01)and increasing time to surgery.CONCLUSION Our results do not support the hypothesis that increased time from injury to isolated meniscus surgery worsens postoperative outcomes.Decision-making primarily based on injury interval is thus not recommended.展开更多
Erythroid cells, the predominant circulating blood cells, are essential for oxygen and carbon dioxide transport (Obeagu, 2024).Their production, erythropoiesis, involves the coordinated synthesis of globin chains and ...Erythroid cells, the predominant circulating blood cells, are essential for oxygen and carbon dioxide transport (Obeagu, 2024).Their production, erythropoiesis, involves the coordinated synthesis of globin chains and heme molecules to assemble hemoglobin(Zhang et al., 2021). The erythroid-specific enzyme δ-aminolevulinate synthase 2 (ALAS2) is a key rate-limiting factor in heme biosynthesis,with its expression increasing in late-stage erythropoiesis to meet heme demands (Sadlon et al., 1999). Zebrafish (Danio rerio) is a well-established model for studying erythropoiesis due to its genetic tractability and optical transparency (Zhang and Hamza, 2019;Zhang et al., 2021). The Tol2-mediated Gal4-UAS system has been widely applied for gene and enhancer trapping in zebrafish (Asakawa and Kawakami, 2009). However, reliable Gal4 enhancer-trap lines for erythropoiesis remain limited. Here, we report a transgenic zebrafish line with erythroid-specific Gal4FF expression under the control of the endogenous alas2 promoter, offering a more precise erythroblast labeling than the gata1a reporter line. This model provides a valuable tool for erythroid-specific investigations of blood flow dynamics and gene function.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
BACKGROUND Colonic diverticular bleeding is a significant cause of acute overt lower gastrointestinal hemorrhage.Colonoscopy is commonly performed in patients with colonic diverticular bleeding to identify and treat t...BACKGROUND Colonic diverticular bleeding is a significant cause of acute overt lower gastrointestinal hemorrhage.Colonoscopy is commonly performed in patients with colonic diverticular bleeding to identify and treat the source of bleeding,which is crucial for effective management.CASE SUMMARY We present the case of a 70-year-old male with acute painless hematochezia.Colonoscopy showed active bleeding originating from a diverticulum,and the EyeMax direct visualization system was used in the treatment of the colonic diverticular bleeding.No further evidence of bleeding was observed at the end of the procedure.The patient was discharged from hospital on the seventh day.CONCLUSION The EyeMax system proved advantageous in localizing the bleeding diverticulum,thereby facilitating successful hemostatic measures.展开更多
The advancement of tissue clearing technology has significantly propelled neuroscience research.Nevertheless,the fluorescent proteins used in traditional transgenic mouse strains were not specifically optimized for ti...The advancement of tissue clearing technology has significantly propelled neuroscience research.Nevertheless,the fluorescent proteins used in traditional transgenic mouse strains were not specifically optimized for tissue clearing procedures,resulting in a substantial decrease in fluorescent intensity after clearing.In this study,we developed the Ci1 reporter mouse strain(where Ci stands for the Chinese Institute for Brain Research,CIBR)based on the bright red fluorescent protein mScarlet.The Ci1 reporter exhibits no fluorescence leakage in various organs or tissue types and can be readily crossed with multiple tissue-specific Cre lines.Compared to the Ai14 mouse strain,the Ci1 reporter strain demonstrates lower non-specific leakage,stronger fluorescence intensity in different tissues,and better preservation of fluorescence following tissue clearing treatment.The creation of the Ci1 reporter provides a more effective tool for both neuroscience and other biomedical research applications.展开更多
BACKGROUND Systemic lupus erythematosus(SLE)can affect multiple organs or systems.The involvement of the central nervous system can result in the manifestation of epilepsy,an acute confusional state,and other rare neu...BACKGROUND Systemic lupus erythematosus(SLE)can affect multiple organs or systems.The involvement of the central nervous system can result in the manifestation of epilepsy,an acute confusional state,and other rare neuropsychiatric presentations,such as catatonia.CASE SUMMARY We present a case of an adolescent male patient with first-onset SLE who presented with neuropsychiatric symptoms including epilepsy and delirium.The initial utilization of olanzapine to alleviate symptoms of agitation precipitated the emergence of catatonia,which was mitigated by discontinuing olanzapine and supplementing with lorazepam.In this case,whether the catatonia was secondary to the utilization of antipsychotics or to an organic disease is a question that warrants differential diagnosis.CONCLUSION Multidisciplinary collaborative management is the cornerstone for the successful management of severe cases of SLE.展开更多
BACKGROUND Transjugular intrahepatic portosystemic shunt(TIPS)is a highly effective treat-ment for complications associated with portal hypertension.However,stent fracture,although extremely rare,represents a potentia...BACKGROUND Transjugular intrahepatic portosystemic shunt(TIPS)is a highly effective treat-ment for complications associated with portal hypertension.However,stent fracture,although extremely rare,represents a potentially serious complication following TIPS creation.Timely identification and management are crucial for preventing further adverse events.CASE SUMMARY We report a 56-year-old male patient who underwent a TIPS procedure for re-current melena caused by portal hypertension secondary to hepatitis B and experienced a stent fracture 15 months post-procedure.He was readmitted 30 months after the initial TIPS due to recurrent esophagogastric variceal bleeding and ascites.An attempt to revise the dysfunctional shunt via a stent-in-stent approach was unsuccessful.Consequently,a parallel TIPS procedure was success-fully performed via the proximal end of the fractured stent to decompress the portal venous system.At the 1-month follow-up,the patient exhibited no recur-rent variceal bleeding,and his ascites had significantly decreased.Twelve-month postoperative monitoring revealed no hepatic encephalopathy and no recurrence of bleeding or ascites.Additionally,we review the existing literature on post-TIPS stent fractures to explore the underlying mechanisms contributing to this com-plication.CONCLUSION Early recognition and prompt intervention are essential in managing stent fractures after TIPS creation to mitigate potential risks and ensure optimal patient outcomes.展开更多
Objective This study aimed to assess the local staging of bladder tumors in patients utilizing preoperative multiparametric MRI(mpMRI)and to demonstrate the clinical efficacy of this method through a comparative analy...Objective This study aimed to assess the local staging of bladder tumors in patients utilizing preoperative multiparametric MRI(mpMRI)and to demonstrate the clinical efficacy of this method through a comparative analysis with corresponding histopathological findings.Methods Between November 2020 and April 2022,63 patients with a planned cystoscopy and a preliminary or previous diagnosis of bladder tumor were included.All participants underwent mpMRI,and Vesical Imaging Reporting and Data System(VI-RADS)criteria were applied to assess the recorded images.Subsequently,obtained biopsies were histopathologically examined and compared with radiological findings.Results Of the 63 participants,60 were male,and three were female.Categorizing tumors with a VI-RADS score of>3 as muscle invasive,84%were radiologically classified as having an invasive bladder tumor.However,histopathological results indicated invasive bladder tumors in 52%of cases.Sensitivity of the VI-RADS score was 100%;specificity was 23%;the negative predictive value was 100%;and the positive predictive value was 62%.Conclusion The scoring system obtained through mpMRI,VI-RADS,proves to be a successful method,particularly in determining the absence of muscle invasion in bladder cancer.Its efficacy in detecting muscle invasion in bladder tumors could be further enhanced with additional studies,suggesting potential for increased diagnostic efficiency through ongoing research.The VI-RADS could enhance the selection of patients eligible for accurate diagnosis and treatment.展开更多
Background:Lenvatinib is primarily utilized for the treatment of inoperable or advanced hepatocellular carcinoma,radioiodine-refractory differentiated thyroid cancer,and advanced renal cell carcinoma.Primary informati...Background:Lenvatinib is primarily utilized for the treatment of inoperable or advanced hepatocellular carcinoma,radioiodine-refractory differentiated thyroid cancer,and advanced renal cell carcinoma.Primary information about adverse reactions is principally derived from clinical trials;however,there is a notable dearth of substantial real-world studies.Methods:In this research,an examination of the U.S.Food and Drug Administration(FDA)Adverse Event Reporting System(FAERS)database was performed to evaluate the potential side effects of lenvatinib.The FAERS database revealed a total of 20,290 reported adverse events associated with lenvatinib.Different algorithms for repeated measure analysis were employed to ascertain the significance of these adverse reactions.Results:The study identified 170 instances of adverse events(AEs)induced by lenvatinib,incorporating several significant adverse reactions that the product label does not mention.The investigation also evaluated the onset periods of the adverse reactions,pinpointing a median time of 43 days.The majority of adverse reactions manifested within the initial month of lenvatinib use.Sex-specific analysis revealed disparities in high risk adverse reactions between females(vascular and lymphatic diseases,and neuronal organ diseases)and males(death and infectious diseases).Our data mining has unveiled adverse reactions beyond those mentioned within the instructions,such as osteonecrosis of the jaw,cholecystitis,cholangitis,dehydration,tumor lysis syndrome,type 1 diabetes,hyperammonemia,liver abscess,interstitial lung disease,pneumothorax,sudden death,and aortic dissection.Conclusion:The insights derived from these findings contribute significant nuances for optimizing lenvatinib use,enhancing its efficacy,and substantially mitigating potential side effects.These data elements will substantially enhance the implementation of the drug in a clinical environment.展开更多
BACKGROUND Liver cirrhosis patients can develop various complications including bacteremia as the intestinal flora is heterologous.In those with low immunity,trauma,or following surgery,etc.,the body is susceptible to...BACKGROUND Liver cirrhosis patients can develop various complications including bacteremia as the intestinal flora is heterologous.In those with low immunity,trauma,or following surgery,etc.,the body is susceptible to concurrent systemic or local infections.Under these circumstances,even minimally invasive treatment methods such as interventional therapy like transjugular intrahepatic portosystemic shunt(TIPS)for liver cirrhosis patients can also result in complications such as infections.CASE SUMMARY A male patient with decompensated cirrhosis experienced multiple episodes of gastrointestinal bleeding and hypersplenism.He was admitted to hospital due to voluntary remedial TIPS.The patient developed a numerous intrahepatic liver abscess postoperatively.Following initial conservative treatment with intravenous antibiotics and parenteral nutrition,three months after TIPS,the liver abscess had disappeared on imaging examination.At the 6-month postoperative follow-up,outpatient re-examination showed that the patient had recovered and the liver abscess had resolved.CONCLUSION Attention should be paid to decreased blood cell counts,especially low leukocyte levels in patients with liver cirrhosis as the presence of intestinal microbiota dysregulation and portal pyemia can result in liver abscess and sepsis during invasive diagnostic and therapeutic procedures like TIPS.The addition of probiotics might reduce the risk in such patients.展开更多
BACKGROUND Angiostrongylus cantonensis-induced acute parasitic infection is a rare food-borne disease in clinical practice.Lack of its specific laboratory markers and subsequent difficulty in detecting pathogens cause...BACKGROUND Angiostrongylus cantonensis-induced acute parasitic infection is a rare food-borne disease in clinical practice.Lack of its specific laboratory markers and subsequent difficulty in detecting pathogens cause high misdiagnosis and missed diagnosis rates.CASE SUMMARY A 20-year-old male developed persistent neck and back pain after consuming raw snail meat,followed by urinary retention and low fever.After admission,the patient was misdiagnosed as viral infection and Mycobacterium tuberculosis in central nervous system.After detection of Angiostrongylus cantonensis in blood and cerebrospinal fluid by metagenomics next generation sequencing,albendazole was administered with ceftriaxone and methylprednisolone treatment simultaneously.With effective antiparasitic treatment,the patient weaned from mechanical ventilation successfully and transferred out of intensive care unit for hyperbaric oxygen and rehabilitation treatment.CONCLUSION This case highlights the diagnostic challenges of Angiostrongylus cantonensis infection and the importance of advanced sequencing techniques in identifying rare pathogens.展开更多
BACKGROUND Recent advancements in artificial intelligence(AI)have significantly enhanced the capabilities of endoscopic-assisted diagnosis for gastrointestinal diseases.AI has shown great promise in clinical practice,...BACKGROUND Recent advancements in artificial intelligence(AI)have significantly enhanced the capabilities of endoscopic-assisted diagnosis for gastrointestinal diseases.AI has shown great promise in clinical practice,particularly for diagnostic support,offering real-time insights into complex conditions such as esophageal squamous cell carcinoma.CASE SUMMARY In this study,we introduce a multimodal AI system that successfully identified and delineated a small and flat carcinoma during esophagogastroduodenoscopy,highlighting its potential for early detection of malignancies.The lesion was confirmed as high-grade squamous intraepithelial neoplasia,with pathology results supporting the AI system’s accuracy.The multimodal AI system offers an integrated solution that provides real-time,accurate diagnostic information directly within the endoscopic device interface,allowing for single-monitor use without disrupting endoscopist’s workflow.CONCLUSION This work underscores the transformative potential of AI to enhance endoscopic diagnosis by enabling earlier,more accurate interventions.展开更多
BACKGROUND Adolescent/adult pancreatoblastoma(PB)is an uncommon malignant pancreatic tumor.The paucity of data stemming from the rarity of this disease leads to minimal generalized guidelines regarding its diagnosis a...BACKGROUND Adolescent/adult pancreatoblastoma(PB)is an uncommon malignant pancreatic tumor.The paucity of data stemming from the rarity of this disease leads to minimal generalized guidelines regarding its diagnosis and treatment.There is a limited number of case reports in the literature and there has been no recent analysis of the literature to consolidate their common features.The purpose of the featured study is to review the available cases of adolescent/adult PB and analyze the common genetic features,histologic features,treatment regimens,tumor sizes,tumor locations,and areas of metastasis to advance ongoing research and better understand and treat this rare condition.AIM To present a patient case and systematically review all available cases in the literature to consolidate the common physical,genetic,and histologic features of PB.METHODS This is a systematic review of the literature with a case study.A total of 89 patient cases were discovered in the literature database for adolescent/adult PB,all of which were reviewed and are included in our research.Patients aged 16-18 were considered adolescent and patients aged greater than 18 were considered adult.Adolescents and adults were grouped together for the purpose of this study.The patient from the case report was seen in a community hospital setting.RESULTS The 89 cases analyzed from the literature were found in 51 references(our case report included),which were consolidated into the six categories mentioned above.A plurality of references reports PB in the head of the pancreas,4.0-10.0 cm in size,and with the most common site of metastasis to the liver.Histology studies most commonly included acinar groups,squamous corpuscles/nests,cytokeratin,chromogranin,trypsin,chymotrypsin,and synaptophysin.Genetic studies most commonly included adenomatous polyposis coli,B-cell lymphoma/leukemia 10,catenin beta 1,and Wnt/beta-catenin mutations.The mainstay of treatment was surgery with chemotherapy typically including cisplatin,carboplatin,doxorubicin,5 fluorouracil,mitomycin,bleomycin,gemcitabine,and vindesine.Radiation was also often used.CONCLUSION Common pancreatoblastoma features include acinar groups,chromogranin,chymotrypsin,squamous corpuscles,synaptophysin and trypsin on histology and adenomatous polyposis coli,B-cell lymphoma/leukemia 10,catenin beta 1,and Wnt/beta-catenin genetic mutations.展开更多
BACKGROUND Recently,there has been a significant increase in the consumption of ultraprocessed foods worldwide.However,the association between the consumption of ultra-processed food,obesity,and the prevalence of colo...BACKGROUND Recently,there has been a significant increase in the consumption of ultraprocessed foods worldwide.However,the association between the consumption of ultra-processed food,obesity,and the prevalence of colon cancer remains controversial.AIM To find out the association between the consumption of ultra-processed food,obesity,and the prevalence of colon cancer.METHODS A comprehensive systematic literature search of PubMed,Scopus,Web of Science,and Google Scholar for grey literature was done for articles published before 8th March 2023.The search was done to retrieve potential peer-reviewed articles that explored the association between the consumption of ultra-processed food,obesity,and the prevalence of colon cancer.RESULTS Of the 246 potential articles assessed,17 met the inclusion criteria.Meta-analysis results demonstrated that high consumption of ultra-processed food is associated with an increased risk of obesity[odds ratio(OR):1.65;95%CI:1.07-2.45;P<0.05].Consequently,there is a positive association between obesity and an increased risk of colon cancer(OR 1.48;95%CI:0.77-2.87;P>0.05).CONCLUSION Consuming ultra-processed foods increases the risk of obesity and colon cancer.展开更多
Cancer stem cells(CSCs)are tumor cells that share functional characteristics with normal and embryonic stem cells.CSCs have increased tumor-initiating capacity and metastatic potential and lower sensitivity to chemo-a...Cancer stem cells(CSCs)are tumor cells that share functional characteristics with normal and embryonic stem cells.CSCs have increased tumor-initiating capacity and metastatic potential and lower sensitivity to chemo-and radiotherapy,with important roles in tumor progression and the response to therapy.Thus,a current goal of cancer research is to eliminate CSCs,necessitating an adequate phenotypic and functional characterization of CSCs.Strategies have been developed to identify,enrich,and track CSCs,many of which distinguish CSCs by evaluating the expression of surface markers,the initiation of specific signaling pathways,and the activation of master transcription factors that control stemness in normal cells.We review and discuss the use of reporter gene systems for identifying CSCs.Reporters that are under the control of aldehyde dehydrogenase 1A1,CD133,Notch,Nanog homeobox,Sex-determining region Y-box 2,and POU class 5 homeobox can be used to identify CSCs in many tumor types,track cells in real time,and screen for drugs.Thus,reporter gene systems,in combination with in vitro and in vivo functional assays,can assess changes in the CSCs pool.We present relevant examples of these systems in the evaluation of experimental CSCs-targeting therapeutics,demonstrating their value in CSCs research.展开更多
Zika virus(ZIKV) is associated with severe birth defects and Guillain-Barre′ syndrome and no approved vaccines or specific therapies to combat ZIKV infection are currently available. To accelerate anti-ZIKV therapeut...Zika virus(ZIKV) is associated with severe birth defects and Guillain-Barre′ syndrome and no approved vaccines or specific therapies to combat ZIKV infection are currently available. To accelerate anti-ZIKV therapeutics research, we developed a stable ZIKV GFP-reporter virus system with considerably improved GFP visibility and stability. In this system a BHK-21 cell line expressing DC-SIGNR was established to facilitate the proliferation of GFP-reporter ZIKV. Using this reporter virus system, we established a high-throughput screening assay and screened a selected plant-sourced compounds library for their ability to block ZIKV infection. More than 31 out of 974 tested compounds effectively decreased ZIKV reporter infection. Four selected compounds, homoharringtonine(HHT), bruceine D(BD), dihydroartemisinin(DHA) and digitonin(DGT), were further validated to inhibit wild-type ZIKV infection in cells of BHK-21 and human cell line A549.The FDA-approved chronic myeloid leukemia treatment drug HHT and BD were identified as broad-spectrum flavivirus inhibitors. DHA, another FDA-approved antimalarial drug effectively inhibited ZIKV infection in BHK-21 cells. HHT, BD and DHA inhibited ZIKV infection at a post-entry stage. Digitonin was found to have inhibitory activity in the early stage of viral infection. Our research provides an efficient high-throughput screening assay for ZIKV inhibitors. The active compounds identified in this study represent potential therapies for the treatment of ZIKV infection.展开更多
Objective To develop an alternative method for assessment of gene delivery systems in vivo.Methods Mouse primary spleen lymphocytes were genetically modified in vitro by a retroviral vector harboring a Gaussia lucifer...Objective To develop an alternative method for assessment of gene delivery systems in vivo.Methods Mouse primary spleen lymphocytes were genetically modified in vitro by a retroviral vector harboring a Gaussia luciferase(Gluc) expression cassette.After implantation of these cells into recipient mice,the expression of Gluc was detected in whole blood or plasma collected.Results As little as 10 μL whole blood drawn from the recipient mice could guarantee prompt reading of Gluc activity with a luminometer.And the reading was found in good correlation with the number of genetically modified spleen lymphocytes implanted to the mice.Conclusions Gluc may be useful as an in vivo reporter for gene therapy researches,and Gluc blood assay could provide an alternative method for assessment of gene delivery systems in vivo.展开更多
基金financially supported by the National Natural Science Foundation of China(32470288)the Zhejiang University Global Partnership Fund.
文摘Dear Editor,Understanding the complex interactions between transcription factors(TFs)and DNA is crucial in various scientific fields,including genetics,molecular biology,and biochemistry.Current methodologies such as yeast one-hybrid,chromatin immunoprecipitation followed by sequencing,the firefly luciferase(Luc)reporter system,and DNA electrophoretic mobility shift assays often necessitate complicated procedures and costly chemicals(Ferraz et al.,2021).The adoption of the pigmentation-based reporter RUBY has facilitated the study of various plant signaling processes due to the easily observable nature of pigments(He et al.,2020).However,pigmentation-based assays lack sensitivity,and their high background limits broader applications.The recent discovery of the fungal Luc coding gene Luz in the bioluminescence pathway from the fungus Neonothopanus nambi offers new opportunities for enhancing reporter systems in transcriptional regulation assays(Kotlobay et al.,2018).The fungal bioluminescence pathway(FBP)is a cyclic metabolic route that involves the action of four enzymes:HispS,H3H,Luz,and CPH(Figure 1A).The pathway’s initial committed step utilizes caffeic acid as a substrate by the fungus HispS;NPGA from Aspergillus nidulans is capable of post-translationally modifying HispS.This modification enhances the activity of HispS,thereby increasing the intensity of FBP(Khakhar et al.,2020;Zheng et al.,2023;Shakhova et al.,2024).Notably,the synthesis pathway of caffeic acid has evolutionarily conserved homologs in plants(Supplemental Figure 1),suggesting that plant cells might independently utilize their own metabolic pool of caffeic acid to support the FBP without the need for additional substrate.This finding underscores the potential of FBP as a promising reporter for plant transcriptional regulation without substrate addition.
基金supported by the National Key Research and Development Program of China(award no.2023YFD1202900)the National Science Foundation of China(award nos.32270433 and 32101205)+4 种基金the Natural Science Foundation of Sichuan Province(award no.2022NSFSC0143)to Y.Z.and X.T.,the Joint Science and Technology Project between Sichuan Province and Chongqing Municipality(award no.CSTC2021JSCXCYLHX0001)to H.S.and X.T.the Modern Seed Industry Project of Chongqing Municipal Science and Technology Bureau(award no.CSTB2023TIAD-KPX0025)to H.S.the National Science Foundation of China(award no.32301248)to Q.R.the National Science Foundation of China(award no.32072045)to X.Z.supported by the NSF Plant Genome Research Program(award nos.IOS-2029889 and IOS-2132693)to Y.Q.
文摘CRISPR-Cas-based genome editing holds immense promise for advancing plant genomics and crop enhancement.However,the challenge of low editing activity complicates the identification of editing events.In this study,we introduce multiple single transcript unit surrogate reporter(STU-SR)systems to enhance the selection of genome-edited plants.These systems use the same single guide RNAs designed for endogenous genes to edit reporter genes,establishing a direct link between reporter gene editing activity and that of endogenous genes.Various strategies are used to restore functional reporter genes after genome editing,including efficient single-strand annealing(SSA)for homologous recombination in STUSR-SSA systems.STU-SR-base editor systems leverage base editing to reinstate the start codon,enriching C-to-T and A-to-G base editing events.Our results showcase the effectiveness of these STU-SR systems in enhancing genome editing events in the monocot rice,encompassing Cas9 nuclease-based targeted mutagenesis,cytosine base editing,and adenine base editing.The systems exhibit compatibility with Cas9 variants,such as the PAM-less SpRY,and are shown to boost genome editing in Brassica oleracea,a dicot vegetable crop.In summary,we have developed highly efficient and versatile STU-SR systems for enrichment of genome-edited plants.
文摘BACKGROUND Thrombophilia contributes to a significant increased risk of venous thromboembolism and can be either inherited or acquired.Hereditary thrombophilia may arise from various gene mutations,some of which have not even been adequately reported or poorly understood.Previous studies reported a rare and novel missense mutation in the prothrombin gene(p.Arg596Gln),known as prothrombin Belgrade.The mechanisms and therapeutic strategies associated with prothrombin Belgrade mutation have not been fully elucidated.CASE SUMMARY We present the case of a 26-year-old woman with recurrent systemic thrombosis induced by prothrombin Belgrade mutation.The patient suffered from cerebral venous sinus thrombosis that rapidly progressed to systemic thrombosis,alongside a family history of cerebral thrombosis,and no traditional risk factors or abnormal coagulation function.Whole-genome sequencing detected a novel and rare heterozygous prothrombin missense mutation,c.1787G>T(p.Arg596Gln),which was responsible for the major etiology of the systemic thrombosis.CONCLUSION This case strengthens our understanding about hereditary basis of thrombophilia and provokes considerations for therapeutic options on prothrombin Belgrade mutation.
文摘BACKGROUND Meniscal tears are one of the most common knee injuries.After the diagnosis of a meniscal tear has been made,there are several factors physicians use to guide clinical decision-making.The influence of time between injury and isolated meniscus repair on patient outcomes is not well described.Assessing this relationship is important as it may influence clinical decision-making and can add to the preoperative patient education process.We hypothesized that increasing the time from injury to meniscus surgery would worsen postoperative outcomes.AIM To investigate the current literature for data on the relationship between time between meniscus injury and repair on patient outcomes.METHODS PubMed,Academic Search Complete,MEDLINE,CINAHL,and SPORTDiscus were searched for studies published between January 1,1995 and July 13,2023 on isolated meniscus repair.Exclusion criteria included concomitant ligament surgery,incomplete outcomes or time to surgery data,and meniscectomies.Patient demographics,time to injury,and postoperative outcomes from each study were abstracted and analyzed.RESULTS Five studies met all inclusion and exclusion criteria.There were 204(121 male,83 female)patients included.Three of five(60%)studies determined that time between injury and surgery was not statistically significant for postoperative Lysholm scores(P=0.62),Tegner scores(P=0.46),failure rate(P=0.45,P=0.86),and International Knee Documentation Committee scores(P=0.65).Two of five(40%)studies found a statistically significant increase in Lysholm scores with shorter time to surgery(P=0.03)and a statistically significant association between progression of medial meniscus extrusion ratio(P=0.01)and increasing time to surgery.CONCLUSION Our results do not support the hypothesis that increased time from injury to isolated meniscus surgery worsens postoperative outcomes.Decision-making primarily based on injury interval is thus not recommended.
基金supported by the National Key Research and Development Plan of China (2023YFA1802000)the National Natural Science Foundation of China for Distinguished Young Scholars (31925014),the National Natural Science Foundation of China Key Program (32130033),the National Natural Science Foundation of Original Exploratory Program (32350006)+5 种基金the Key Research Program of Frontier Sciences,Chinese Academy of Sciences (ZDBS-LY-SM010)Shanghai Pilot Program for Basic Research-Chinese Academy of Sciences,Shanghai Branch (JCYJ-SHFY-2022-006)Shanghai Science Technology Innovation Action Plan for Basic Research Program (21JC1406300)Haihe laboratory of Cell Ecosystem Innovation Fund (24HHXBSS00011)CAS project for Young Scientists in Basic Research (YSBR-077)supported by Science and Technology Commission of Shanghai Municipality (Shanghai Rising-Star Program,23QA1411300)
文摘Erythroid cells, the predominant circulating blood cells, are essential for oxygen and carbon dioxide transport (Obeagu, 2024).Their production, erythropoiesis, involves the coordinated synthesis of globin chains and heme molecules to assemble hemoglobin(Zhang et al., 2021). The erythroid-specific enzyme δ-aminolevulinate synthase 2 (ALAS2) is a key rate-limiting factor in heme biosynthesis,with its expression increasing in late-stage erythropoiesis to meet heme demands (Sadlon et al., 1999). Zebrafish (Danio rerio) is a well-established model for studying erythropoiesis due to its genetic tractability and optical transparency (Zhang and Hamza, 2019;Zhang et al., 2021). The Tol2-mediated Gal4-UAS system has been widely applied for gene and enhancer trapping in zebrafish (Asakawa and Kawakami, 2009). However, reliable Gal4 enhancer-trap lines for erythropoiesis remain limited. Here, we report a transgenic zebrafish line with erythroid-specific Gal4FF expression under the control of the endogenous alas2 promoter, offering a more precise erythroblast labeling than the gata1a reporter line. This model provides a valuable tool for erythroid-specific investigations of blood flow dynamics and gene function.
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.
文摘BACKGROUND Colonic diverticular bleeding is a significant cause of acute overt lower gastrointestinal hemorrhage.Colonoscopy is commonly performed in patients with colonic diverticular bleeding to identify and treat the source of bleeding,which is crucial for effective management.CASE SUMMARY We present the case of a 70-year-old male with acute painless hematochezia.Colonoscopy showed active bleeding originating from a diverticulum,and the EyeMax direct visualization system was used in the treatment of the colonic diverticular bleeding.No further evidence of bleeding was observed at the end of the procedure.The patient was discharged from hospital on the seventh day.CONCLUSION The EyeMax system proved advantageous in localizing the bleeding diverticulum,thereby facilitating successful hemostatic measures.
基金supported by the startup funding from the Chinese Institute for Brain Research to Hu Zhao.
文摘The advancement of tissue clearing technology has significantly propelled neuroscience research.Nevertheless,the fluorescent proteins used in traditional transgenic mouse strains were not specifically optimized for tissue clearing procedures,resulting in a substantial decrease in fluorescent intensity after clearing.In this study,we developed the Ci1 reporter mouse strain(where Ci stands for the Chinese Institute for Brain Research,CIBR)based on the bright red fluorescent protein mScarlet.The Ci1 reporter exhibits no fluorescence leakage in various organs or tissue types and can be readily crossed with multiple tissue-specific Cre lines.Compared to the Ai14 mouse strain,the Ci1 reporter strain demonstrates lower non-specific leakage,stronger fluorescence intensity in different tissues,and better preservation of fluorescence following tissue clearing treatment.The creation of the Ci1 reporter provides a more effective tool for both neuroscience and other biomedical research applications.
基金Supported by STI2030-Major Projects,No.2021ZD0202001Capital Funds for Health Improvement and Research,No.CFH 2022-2-4012.
文摘BACKGROUND Systemic lupus erythematosus(SLE)can affect multiple organs or systems.The involvement of the central nervous system can result in the manifestation of epilepsy,an acute confusional state,and other rare neuropsychiatric presentations,such as catatonia.CASE SUMMARY We present a case of an adolescent male patient with first-onset SLE who presented with neuropsychiatric symptoms including epilepsy and delirium.The initial utilization of olanzapine to alleviate symptoms of agitation precipitated the emergence of catatonia,which was mitigated by discontinuing olanzapine and supplementing with lorazepam.In this case,whether the catatonia was secondary to the utilization of antipsychotics or to an organic disease is a question that warrants differential diagnosis.CONCLUSION Multidisciplinary collaborative management is the cornerstone for the successful management of severe cases of SLE.
文摘BACKGROUND Transjugular intrahepatic portosystemic shunt(TIPS)is a highly effective treat-ment for complications associated with portal hypertension.However,stent fracture,although extremely rare,represents a potentially serious complication following TIPS creation.Timely identification and management are crucial for preventing further adverse events.CASE SUMMARY We report a 56-year-old male patient who underwent a TIPS procedure for re-current melena caused by portal hypertension secondary to hepatitis B and experienced a stent fracture 15 months post-procedure.He was readmitted 30 months after the initial TIPS due to recurrent esophagogastric variceal bleeding and ascites.An attempt to revise the dysfunctional shunt via a stent-in-stent approach was unsuccessful.Consequently,a parallel TIPS procedure was success-fully performed via the proximal end of the fractured stent to decompress the portal venous system.At the 1-month follow-up,the patient exhibited no recur-rent variceal bleeding,and his ascites had significantly decreased.Twelve-month postoperative monitoring revealed no hepatic encephalopathy and no recurrence of bleeding or ascites.Additionally,we review the existing literature on post-TIPS stent fractures to explore the underlying mechanisms contributing to this com-plication.CONCLUSION Early recognition and prompt intervention are essential in managing stent fractures after TIPS creation to mitigate potential risks and ensure optimal patient outcomes.
文摘Objective This study aimed to assess the local staging of bladder tumors in patients utilizing preoperative multiparametric MRI(mpMRI)and to demonstrate the clinical efficacy of this method through a comparative analysis with corresponding histopathological findings.Methods Between November 2020 and April 2022,63 patients with a planned cystoscopy and a preliminary or previous diagnosis of bladder tumor were included.All participants underwent mpMRI,and Vesical Imaging Reporting and Data System(VI-RADS)criteria were applied to assess the recorded images.Subsequently,obtained biopsies were histopathologically examined and compared with radiological findings.Results Of the 63 participants,60 were male,and three were female.Categorizing tumors with a VI-RADS score of>3 as muscle invasive,84%were radiologically classified as having an invasive bladder tumor.However,histopathological results indicated invasive bladder tumors in 52%of cases.Sensitivity of the VI-RADS score was 100%;specificity was 23%;the negative predictive value was 100%;and the positive predictive value was 62%.Conclusion The scoring system obtained through mpMRI,VI-RADS,proves to be a successful method,particularly in determining the absence of muscle invasion in bladder cancer.Its efficacy in detecting muscle invasion in bladder tumors could be further enhanced with additional studies,suggesting potential for increased diagnostic efficiency through ongoing research.The VI-RADS could enhance the selection of patients eligible for accurate diagnosis and treatment.
基金The 2022 Educational Teaching Reform and Research Project of Guangxi University of Traditional Chinese Medicine(2022C032).
文摘Background:Lenvatinib is primarily utilized for the treatment of inoperable or advanced hepatocellular carcinoma,radioiodine-refractory differentiated thyroid cancer,and advanced renal cell carcinoma.Primary information about adverse reactions is principally derived from clinical trials;however,there is a notable dearth of substantial real-world studies.Methods:In this research,an examination of the U.S.Food and Drug Administration(FDA)Adverse Event Reporting System(FAERS)database was performed to evaluate the potential side effects of lenvatinib.The FAERS database revealed a total of 20,290 reported adverse events associated with lenvatinib.Different algorithms for repeated measure analysis were employed to ascertain the significance of these adverse reactions.Results:The study identified 170 instances of adverse events(AEs)induced by lenvatinib,incorporating several significant adverse reactions that the product label does not mention.The investigation also evaluated the onset periods of the adverse reactions,pinpointing a median time of 43 days.The majority of adverse reactions manifested within the initial month of lenvatinib use.Sex-specific analysis revealed disparities in high risk adverse reactions between females(vascular and lymphatic diseases,and neuronal organ diseases)and males(death and infectious diseases).Our data mining has unveiled adverse reactions beyond those mentioned within the instructions,such as osteonecrosis of the jaw,cholecystitis,cholangitis,dehydration,tumor lysis syndrome,type 1 diabetes,hyperammonemia,liver abscess,interstitial lung disease,pneumothorax,sudden death,and aortic dissection.Conclusion:The insights derived from these findings contribute significant nuances for optimizing lenvatinib use,enhancing its efficacy,and substantially mitigating potential side effects.These data elements will substantially enhance the implementation of the drug in a clinical environment.
文摘BACKGROUND Liver cirrhosis patients can develop various complications including bacteremia as the intestinal flora is heterologous.In those with low immunity,trauma,or following surgery,etc.,the body is susceptible to concurrent systemic or local infections.Under these circumstances,even minimally invasive treatment methods such as interventional therapy like transjugular intrahepatic portosystemic shunt(TIPS)for liver cirrhosis patients can also result in complications such as infections.CASE SUMMARY A male patient with decompensated cirrhosis experienced multiple episodes of gastrointestinal bleeding and hypersplenism.He was admitted to hospital due to voluntary remedial TIPS.The patient developed a numerous intrahepatic liver abscess postoperatively.Following initial conservative treatment with intravenous antibiotics and parenteral nutrition,three months after TIPS,the liver abscess had disappeared on imaging examination.At the 6-month postoperative follow-up,outpatient re-examination showed that the patient had recovered and the liver abscess had resolved.CONCLUSION Attention should be paid to decreased blood cell counts,especially low leukocyte levels in patients with liver cirrhosis as the presence of intestinal microbiota dysregulation and portal pyemia can result in liver abscess and sepsis during invasive diagnostic and therapeutic procedures like TIPS.The addition of probiotics might reduce the risk in such patients.
基金Supported by the Hainan Provincial Natural Science Foundation of China,No.824MS173 and No.823MS165the Project of Hainan Province Clinical Medical Center.
文摘BACKGROUND Angiostrongylus cantonensis-induced acute parasitic infection is a rare food-borne disease in clinical practice.Lack of its specific laboratory markers and subsequent difficulty in detecting pathogens cause high misdiagnosis and missed diagnosis rates.CASE SUMMARY A 20-year-old male developed persistent neck and back pain after consuming raw snail meat,followed by urinary retention and low fever.After admission,the patient was misdiagnosed as viral infection and Mycobacterium tuberculosis in central nervous system.After detection of Angiostrongylus cantonensis in blood and cerebrospinal fluid by metagenomics next generation sequencing,albendazole was administered with ceftriaxone and methylprednisolone treatment simultaneously.With effective antiparasitic treatment,the patient weaned from mechanical ventilation successfully and transferred out of intensive care unit for hyperbaric oxygen and rehabilitation treatment.CONCLUSION This case highlights the diagnostic challenges of Angiostrongylus cantonensis infection and the importance of advanced sequencing techniques in identifying rare pathogens.
基金Supported by the 135 High-end Talent Project of West China Hospital,Sichuan University,No.ZYDG23029.
文摘BACKGROUND Recent advancements in artificial intelligence(AI)have significantly enhanced the capabilities of endoscopic-assisted diagnosis for gastrointestinal diseases.AI has shown great promise in clinical practice,particularly for diagnostic support,offering real-time insights into complex conditions such as esophageal squamous cell carcinoma.CASE SUMMARY In this study,we introduce a multimodal AI system that successfully identified and delineated a small and flat carcinoma during esophagogastroduodenoscopy,highlighting its potential for early detection of malignancies.The lesion was confirmed as high-grade squamous intraepithelial neoplasia,with pathology results supporting the AI system’s accuracy.The multimodal AI system offers an integrated solution that provides real-time,accurate diagnostic information directly within the endoscopic device interface,allowing for single-monitor use without disrupting endoscopist’s workflow.CONCLUSION This work underscores the transformative potential of AI to enhance endoscopic diagnosis by enabling earlier,more accurate interventions.
基金Supported by the Honor Health Foundation,Michael and Mary Ellen Francisthe Seena Magowitz Foundation.
文摘BACKGROUND Adolescent/adult pancreatoblastoma(PB)is an uncommon malignant pancreatic tumor.The paucity of data stemming from the rarity of this disease leads to minimal generalized guidelines regarding its diagnosis and treatment.There is a limited number of case reports in the literature and there has been no recent analysis of the literature to consolidate their common features.The purpose of the featured study is to review the available cases of adolescent/adult PB and analyze the common genetic features,histologic features,treatment regimens,tumor sizes,tumor locations,and areas of metastasis to advance ongoing research and better understand and treat this rare condition.AIM To present a patient case and systematically review all available cases in the literature to consolidate the common physical,genetic,and histologic features of PB.METHODS This is a systematic review of the literature with a case study.A total of 89 patient cases were discovered in the literature database for adolescent/adult PB,all of which were reviewed and are included in our research.Patients aged 16-18 were considered adolescent and patients aged greater than 18 were considered adult.Adolescents and adults were grouped together for the purpose of this study.The patient from the case report was seen in a community hospital setting.RESULTS The 89 cases analyzed from the literature were found in 51 references(our case report included),which were consolidated into the six categories mentioned above.A plurality of references reports PB in the head of the pancreas,4.0-10.0 cm in size,and with the most common site of metastasis to the liver.Histology studies most commonly included acinar groups,squamous corpuscles/nests,cytokeratin,chromogranin,trypsin,chymotrypsin,and synaptophysin.Genetic studies most commonly included adenomatous polyposis coli,B-cell lymphoma/leukemia 10,catenin beta 1,and Wnt/beta-catenin mutations.The mainstay of treatment was surgery with chemotherapy typically including cisplatin,carboplatin,doxorubicin,5 fluorouracil,mitomycin,bleomycin,gemcitabine,and vindesine.Radiation was also often used.CONCLUSION Common pancreatoblastoma features include acinar groups,chromogranin,chymotrypsin,squamous corpuscles,synaptophysin and trypsin on histology and adenomatous polyposis coli,B-cell lymphoma/leukemia 10,catenin beta 1,and Wnt/beta-catenin genetic mutations.
文摘BACKGROUND Recently,there has been a significant increase in the consumption of ultraprocessed foods worldwide.However,the association between the consumption of ultra-processed food,obesity,and the prevalence of colon cancer remains controversial.AIM To find out the association between the consumption of ultra-processed food,obesity,and the prevalence of colon cancer.METHODS A comprehensive systematic literature search of PubMed,Scopus,Web of Science,and Google Scholar for grey literature was done for articles published before 8th March 2023.The search was done to retrieve potential peer-reviewed articles that explored the association between the consumption of ultra-processed food,obesity,and the prevalence of colon cancer.RESULTS Of the 246 potential articles assessed,17 met the inclusion criteria.Meta-analysis results demonstrated that high consumption of ultra-processed food is associated with an increased risk of obesity[odds ratio(OR):1.65;95%CI:1.07-2.45;P<0.05].Consequently,there is a positive association between obesity and an increased risk of colon cancer(OR 1.48;95%CI:0.77-2.87;P>0.05).CONCLUSION Consuming ultra-processed foods increases the risk of obesity and colon cancer.
基金Supported by UNAM-PAPIIT,No.IN219719 and No.IA205421CONACYT,No.A1-S-18285.
文摘Cancer stem cells(CSCs)are tumor cells that share functional characteristics with normal and embryonic stem cells.CSCs have increased tumor-initiating capacity and metastatic potential and lower sensitivity to chemo-and radiotherapy,with important roles in tumor progression and the response to therapy.Thus,a current goal of cancer research is to eliminate CSCs,necessitating an adequate phenotypic and functional characterization of CSCs.Strategies have been developed to identify,enrich,and track CSCs,many of which distinguish CSCs by evaluating the expression of surface markers,the initiation of specific signaling pathways,and the activation of master transcription factors that control stemness in normal cells.We review and discuss the use of reporter gene systems for identifying CSCs.Reporters that are under the control of aldehyde dehydrogenase 1A1,CD133,Notch,Nanog homeobox,Sex-determining region Y-box 2,and POU class 5 homeobox can be used to identify CSCs in many tumor types,track cells in real time,and screen for drugs.Thus,reporter gene systems,in combination with in vitro and in vivo functional assays,can assess changes in the CSCs pool.We present relevant examples of these systems in the evaluation of experimental CSCs-targeting therapeutics,demonstrating their value in CSCs research.
基金partially supported by the National Key R&D Program of China (grant 2018YFC1200602 and 2016YFD0500403 to RHH)。
文摘Zika virus(ZIKV) is associated with severe birth defects and Guillain-Barre′ syndrome and no approved vaccines or specific therapies to combat ZIKV infection are currently available. To accelerate anti-ZIKV therapeutics research, we developed a stable ZIKV GFP-reporter virus system with considerably improved GFP visibility and stability. In this system a BHK-21 cell line expressing DC-SIGNR was established to facilitate the proliferation of GFP-reporter ZIKV. Using this reporter virus system, we established a high-throughput screening assay and screened a selected plant-sourced compounds library for their ability to block ZIKV infection. More than 31 out of 974 tested compounds effectively decreased ZIKV reporter infection. Four selected compounds, homoharringtonine(HHT), bruceine D(BD), dihydroartemisinin(DHA) and digitonin(DGT), were further validated to inhibit wild-type ZIKV infection in cells of BHK-21 and human cell line A549.The FDA-approved chronic myeloid leukemia treatment drug HHT and BD were identified as broad-spectrum flavivirus inhibitors. DHA, another FDA-approved antimalarial drug effectively inhibited ZIKV infection in BHK-21 cells. HHT, BD and DHA inhibited ZIKV infection at a post-entry stage. Digitonin was found to have inhibitory activity in the early stage of viral infection. Our research provides an efficient high-throughput screening assay for ZIKV inhibitors. The active compounds identified in this study represent potential therapies for the treatment of ZIKV infection.
基金Supported by National High Technology Research and Development Program of China (863 Program) (2007AA021206,2007AA021106)
文摘Objective To develop an alternative method for assessment of gene delivery systems in vivo.Methods Mouse primary spleen lymphocytes were genetically modified in vitro by a retroviral vector harboring a Gaussia luciferase(Gluc) expression cassette.After implantation of these cells into recipient mice,the expression of Gluc was detected in whole blood or plasma collected.Results As little as 10 μL whole blood drawn from the recipient mice could guarantee prompt reading of Gluc activity with a luminometer.And the reading was found in good correlation with the number of genetically modified spleen lymphocytes implanted to the mice.Conclusions Gluc may be useful as an in vivo reporter for gene therapy researches,and Gluc blood assay could provide an alternative method for assessment of gene delivery systems in vivo.
