Flow cytometry(FCM),characterized by its simplicity,rapid processing,multiparameter analysis,and high sen-sitivity,is widely used in the diagnosis,treatment,and prognosis of hematological malignancies.FCM testing of t...Flow cytometry(FCM),characterized by its simplicity,rapid processing,multiparameter analysis,and high sen-sitivity,is widely used in the diagnosis,treatment,and prognosis of hematological malignancies.FCM testing of tissue samples not only aids in diagnosing and classifying hematological cancers,but also enables the detection of solid tumors.Its ability to detect numerous marker parameters from small samples is particularly useful when dealing with limited cell quantities,such as in fine-needle biopsy samples.This attribute not only addresses the challenge posed by small sample sizes,but also boosts the sensitivity of tumor cell detection.The significance of FCM in clinical and pathological applications continues to grow.To standardize the use of FCM in detecting hematological malignant cells in tissue samples and to improve quality control during the detection process,experts from the Cell Analysis Professional Committee of the Chinese Society of Biotechnology jointly drafted and agreed upon this consensus.This consensus was formulated based on current literature and clinical practices of all experts across clinical,laboratory,and pathological fields in China.It outlines a comprehensive workflow of FCM-based assay for the detection of hematological malignancies in tissue samples,including report content,interpretation,quality control,and key considerations.Additionally,it provides recommendations on antibody panel designs and analytical approaches to enhancing FCM tests,particularly in cases with limited sample sizes.展开更多
We developed a small-tissue extraction device(sTED),an automated system that integrates 1-min mechanical dissociation and enzymatic digestion to extract viable primary cells from ultrasmall tissue samples(5-20 mg)with...We developed a small-tissue extraction device(sTED),an automated system that integrates 1-min mechanical dissociation and enzymatic digestion to extract viable primary cells from ultrasmall tissue samples(5-20 mg)within 10 min.Unlike conventional methods,sTED minimizes cell loss and enhances reproducibility,achieving>90%cell viability in mouse tissues and>60%in human tumors,with 1.5×10^(4)-2.5×10^(4)cells/mg yield from mouse liver.Tailored for biopsies and ultrasmall samples,sTED addresses critical standardization challenges in organoid-based research.展开更多
Accurate discrimination of cell subtypes at the molecular level is especially important for cancer diagnosis,but no current method allows rapid and precise detection of breast cancer subtypes.Herein,we developed an el...Accurate discrimination of cell subtypes at the molecular level is especially important for cancer diagnosis,but no current method allows rapid and precise detection of breast cancer subtypes.Herein,we developed an elegant DNA walker for direct and rapid differentiation of breast cancer cell subtypes via detection of dual-miRNAs in clinical tissue samples.This DNA nanomachine can be specifically initiated by endogenous miR-21 and miR-31,and the sensitivity was dramatically improved due to the DNAzyme-mediated signal amplification.This DNA walker enabled rapid detection of double miRNA characteristics in different breast cell lines and also distinguished the fluctuations in a single cell.Applications of this DNAzyme-based nanomachine in vivo and in clinical samples were demonstrated for efficient detection of breast cancer subtypes,making the method generally applicable for precise management of cancers.展开更多
BACKGROUND Pancreatic ductal leaks complicated by endoscopic ultrasonography-guided tissue sampling(EUS-TS)can manifest as acute pancreatitis.CASE SUMMARY A 63-year-old man presented with persistent abdominal pain and...BACKGROUND Pancreatic ductal leaks complicated by endoscopic ultrasonography-guided tissue sampling(EUS-TS)can manifest as acute pancreatitis.CASE SUMMARY A 63-year-old man presented with persistent abdominal pain and weight loss.Diagnosis:Laboratory findings revealed elevated carbohydrate antigen 19-9(5920 U/mL)and carcinoembryonic antigen(23.7 ng/mL)levels.Magnetic resonance imaging of the pancreas revealed an approximately 3 cm ill-defined space-occupying lesion in the inferior aspect of the head,with severe encasement of the superior mesenteric artery.Pancreatic ductal adenocarcinoma was confirmed after pathological examination of specimens obtained by EUS-TS using the fanning method.Interventions and outcomes:The following day,the patient experienced severe abdominal pain with high amylase(265 U/L)and lipase(1173 U/L)levels.Computed tomography of the abdomen revealed edematous wall thickening of the second portion of the duodenum with adjacent fluid collections and a suspicious leak from either the distal common bile duct or the main pancreatic duct in the head.Endoscopic retrograde cholangiopancreatography revealed dye leakage in the head of the main pancreatic duct.Therefore,a 5F 7 cm linear plastic stent was deployed into the pancreatic duct to divert the pancreatic juice.The patient’s abdominal pain improved immediately after pancreatic stent insertion,and amylase and lipase levels normalized within a week.Neoadjuvant chemotherapy was then initiated.CONCLUSION Using the fanning method in EUS-TS can inadvertently cause damage to the pancreatic duct and may lead to clinically significant pancreatitis.Placing a pancreatic stent may immediately resolve acute pancreatitis and shorten the waiting time for curative therapy.When using the fanning method during EUSTS,ductal structures should be excluded to prevent pancreatic ductal leakage.展开更多
Over the last twenty years,with the development of gene-driven therapies,numerous new drugs have entered clinical use.Very few of these new drugs are suitable for a large number of patients,and all require molecular g...Over the last twenty years,with the development of gene-driven therapies,numerous new drugs have entered clinical use.Very few of these new drugs are suitable for a large number of patients,and all require molecular genetic testing.In lung cancer,gene-targeted therapy has evolved rapidly and has placed demands on the development of diagnostics and tissue sample preparation and logistics.Rapid diagnosis and prevalence assessment are necessary to determine the prognosis of a lung cancer patient based on the latest research findings.Therefore,the molecular-genetic diagnostic pathway must also be accelerated and matured to do the necessary analyses on small samples.Because lung cancer rebiopsy can be difficult,liquid biopsy techniques should be developed to cover more of the treatable mutations.There are obstacles related to tissue sampling,new genomic techniques and access to gene-driven cancer drugs,including their affordability.With this review and case study,we go into the obstacles faced by our clinic and discuss how to tackle these obstacles in lung cancer.We use lung cancer as an example due to its complexity,though these same obstacles are found in different cancers on a minor scale.展开更多
Determining etiology of biliary strictures may be a diagnostic challenge involving multiple procedures and ensuing delays in therapeutic management(1).Delays in diagnosis have been shown to have an impact on survival;...Determining etiology of biliary strictures may be a diagnostic challenge involving multiple procedures and ensuing delays in therapeutic management(1).Delays in diagnosis have been shown to have an impact on survival;indeed,the 5-year survival rate for cholangiocarcinoma is 10%,and early diagnosis is correlated with an increase in survival rate(1).展开更多
Background:Tissue specimens for nasopharyngeal carcinoma(NPC)research are scarce because of sampling dif-ficulties.Previous studies have suggested non-invasive nasopharyngeal brushing as an effective sampling method f...Background:Tissue specimens for nasopharyngeal carcinoma(NPC)research are scarce because of sampling dif-ficulties.Previous studies have suggested non-invasive nasopharyngeal brushing as an effective sampling method for NPC diagnosis.The present study aimed to evaluate the feasibility of nasopharyngeal brushing in the acquisition of NPC nucleic acids for research.Methods:Nasopharyngeal brushing samples were acquired from 24 healthy individuals and 48 NPC patients.Tissues from 48 NPC and 18 nasopharyngitis patients were collected by endoscopic biopsy.The expression levels of tumor suppressor genes(TSGs)and Epstein-Barr virus(EBV)-encoded microRNAs as well as EBV DNA copy number were measured by quantitative polymerase chain reaction in both types of samples.Results:Among six TSGs examined,the expression levels of two genes were significantly decreased in nasopharyn-geal brushing and tissue samples from NPC patients as compared with those from healthy/nasopharyngitis indi-viduals.Four EBV-encoded microRNAs,mir-bart1-5p,mir-bart5,mir-bart6-5p,and mir-bart17-5p,were significantly up-regulated in both NPC brushing and tissue samples compared with those from healthy/nasopharyngitis controls(P<0.001).EBV DNA was significantly increased in both nasopharyngeal brushing samples(P<0.001)and tissue sam-ples(P<0.001)from NPC patients in comparison with those from healthy controls.Conclusions:Nasopharyngeal brushing can obtain sufficient tumoral materials for the analysis of viral nucleic acid,including EBV-encoded microRNAs and EBV DNA.For the detection of TSG expression,nasopharyngeal brushings was feasible but inferior to tissue samples.This study confirms nasopharyngeal brushing as an applicable sampling method that can aid in nucleic acid-based NPC research.展开更多
Proteomics has evolved into a powerful and widely used bioanalytical technique in the study of cancer,especially hepatocellular carcinoma (HCC).In this review,we provide an up to date overview of feasible proteome-ana...Proteomics has evolved into a powerful and widely used bioanalytical technique in the study of cancer,especially hepatocellular carcinoma (HCC).In this review,we provide an up to date overview of feasible proteome-analytical techniques for clinical questions.In addition,we present a broad summaryof proteornic studies of HCC utilizing various technical approaches for the analysis of samples derived from diverse sources like HCC cell lines,animal models,human tissue and body fluids.展开更多
基金supported by grants from the National Natural Science Foundation of China(grant numbers:82370195,82270203,81770211)the Fundamental Research Funds for the Central Univer-sities(grant number:2022CDJYGRH-001)Chongqing Technology Innovation and Application Development Special Key Project(grant number:CSTB2024TIAD-KPX0031).
文摘Flow cytometry(FCM),characterized by its simplicity,rapid processing,multiparameter analysis,and high sen-sitivity,is widely used in the diagnosis,treatment,and prognosis of hematological malignancies.FCM testing of tissue samples not only aids in diagnosing and classifying hematological cancers,but also enables the detection of solid tumors.Its ability to detect numerous marker parameters from small samples is particularly useful when dealing with limited cell quantities,such as in fine-needle biopsy samples.This attribute not only addresses the challenge posed by small sample sizes,but also boosts the sensitivity of tumor cell detection.The significance of FCM in clinical and pathological applications continues to grow.To standardize the use of FCM in detecting hematological malignant cells in tissue samples and to improve quality control during the detection process,experts from the Cell Analysis Professional Committee of the Chinese Society of Biotechnology jointly drafted and agreed upon this consensus.This consensus was formulated based on current literature and clinical practices of all experts across clinical,laboratory,and pathological fields in China.It outlines a comprehensive workflow of FCM-based assay for the detection of hematological malignancies in tissue samples,including report content,interpretation,quality control,and key considerations.Additionally,it provides recommendations on antibody panel designs and analytical approaches to enhancing FCM tests,particularly in cases with limited sample sizes.
基金supported by the National Natural Science Foundation of China(Nos.32371470 and 82341019)the Department of Science and Technology of Guangdong Province(No.2023B0909020003).
文摘We developed a small-tissue extraction device(sTED),an automated system that integrates 1-min mechanical dissociation and enzymatic digestion to extract viable primary cells from ultrasmall tissue samples(5-20 mg)within 10 min.Unlike conventional methods,sTED minimizes cell loss and enhances reproducibility,achieving>90%cell viability in mouse tissues and>60%in human tumors,with 1.5×10^(4)-2.5×10^(4)cells/mg yield from mouse liver.Tailored for biopsies and ultrasmall samples,sTED addresses critical standardization challenges in organoid-based research.
基金This work was supported by the National Natural Science Foundation of China(grant no.21974125)the Program for Science and Technology Innovation Teams in Universities of Henan Province(grant no.22IRTSTHN002)+3 种基金the Key Project of Science and Technology of Henan Province(grant no.212102310334)111 Project of Henan Province(grant no.CXJD2021001)the Collaborative Innovation Project of Zhengzhou(grant no.18XTZX12002)Special Funds for the Construction of Innovative Provinces in Hunan Province(grant no.2019RS1031).
文摘Accurate discrimination of cell subtypes at the molecular level is especially important for cancer diagnosis,but no current method allows rapid and precise detection of breast cancer subtypes.Herein,we developed an elegant DNA walker for direct and rapid differentiation of breast cancer cell subtypes via detection of dual-miRNAs in clinical tissue samples.This DNA nanomachine can be specifically initiated by endogenous miR-21 and miR-31,and the sensitivity was dramatically improved due to the DNAzyme-mediated signal amplification.This DNA walker enabled rapid detection of double miRNA characteristics in different breast cell lines and also distinguished the fluctuations in a single cell.Applications of this DNAzyme-based nanomachine in vivo and in clinical samples were demonstrated for efficient detection of breast cancer subtypes,making the method generally applicable for precise management of cancers.
文摘BACKGROUND Pancreatic ductal leaks complicated by endoscopic ultrasonography-guided tissue sampling(EUS-TS)can manifest as acute pancreatitis.CASE SUMMARY A 63-year-old man presented with persistent abdominal pain and weight loss.Diagnosis:Laboratory findings revealed elevated carbohydrate antigen 19-9(5920 U/mL)and carcinoembryonic antigen(23.7 ng/mL)levels.Magnetic resonance imaging of the pancreas revealed an approximately 3 cm ill-defined space-occupying lesion in the inferior aspect of the head,with severe encasement of the superior mesenteric artery.Pancreatic ductal adenocarcinoma was confirmed after pathological examination of specimens obtained by EUS-TS using the fanning method.Interventions and outcomes:The following day,the patient experienced severe abdominal pain with high amylase(265 U/L)and lipase(1173 U/L)levels.Computed tomography of the abdomen revealed edematous wall thickening of the second portion of the duodenum with adjacent fluid collections and a suspicious leak from either the distal common bile duct or the main pancreatic duct in the head.Endoscopic retrograde cholangiopancreatography revealed dye leakage in the head of the main pancreatic duct.Therefore,a 5F 7 cm linear plastic stent was deployed into the pancreatic duct to divert the pancreatic juice.The patient’s abdominal pain improved immediately after pancreatic stent insertion,and amylase and lipase levels normalized within a week.Neoadjuvant chemotherapy was then initiated.CONCLUSION Using the fanning method in EUS-TS can inadvertently cause damage to the pancreatic duct and may lead to clinically significant pancreatitis.Placing a pancreatic stent may immediately resolve acute pancreatitis and shorten the waiting time for curative therapy.When using the fanning method during EUSTS,ductal structures should be excluded to prevent pancreatic ductal leakage.
文摘Over the last twenty years,with the development of gene-driven therapies,numerous new drugs have entered clinical use.Very few of these new drugs are suitable for a large number of patients,and all require molecular genetic testing.In lung cancer,gene-targeted therapy has evolved rapidly and has placed demands on the development of diagnostics and tissue sample preparation and logistics.Rapid diagnosis and prevalence assessment are necessary to determine the prognosis of a lung cancer patient based on the latest research findings.Therefore,the molecular-genetic diagnostic pathway must also be accelerated and matured to do the necessary analyses on small samples.Because lung cancer rebiopsy can be difficult,liquid biopsy techniques should be developed to cover more of the treatable mutations.There are obstacles related to tissue sampling,new genomic techniques and access to gene-driven cancer drugs,including their affordability.With this review and case study,we go into the obstacles faced by our clinic and discuss how to tackle these obstacles in lung cancer.We use lung cancer as an example due to its complexity,though these same obstacles are found in different cancers on a minor scale.
文摘Determining etiology of biliary strictures may be a diagnostic challenge involving multiple procedures and ensuing delays in therapeutic management(1).Delays in diagnosis have been shown to have an impact on survival;indeed,the 5-year survival rate for cholangiocarcinoma is 10%,and early diagnosis is correlated with an increase in survival rate(1).
基金supported by the Major International Cooperation Projects of the National Natural Science Foundation of China(No.81220108022)the National Natural Science Funds for Distinguished Young Scholars(No.81325018)+3 种基金the National Key Research and Development Program(No.2016YFC1302704)Guangdong Special Support Programs for High-level Personnel(No.2014TX01R201)the Science and Technology Project of Guangdong Province(No.2014B050504004)the Health&Medical Collaborative Innovation Project of Guangzhou City(No.201604020003).
文摘Background:Tissue specimens for nasopharyngeal carcinoma(NPC)research are scarce because of sampling dif-ficulties.Previous studies have suggested non-invasive nasopharyngeal brushing as an effective sampling method for NPC diagnosis.The present study aimed to evaluate the feasibility of nasopharyngeal brushing in the acquisition of NPC nucleic acids for research.Methods:Nasopharyngeal brushing samples were acquired from 24 healthy individuals and 48 NPC patients.Tissues from 48 NPC and 18 nasopharyngitis patients were collected by endoscopic biopsy.The expression levels of tumor suppressor genes(TSGs)and Epstein-Barr virus(EBV)-encoded microRNAs as well as EBV DNA copy number were measured by quantitative polymerase chain reaction in both types of samples.Results:Among six TSGs examined,the expression levels of two genes were significantly decreased in nasopharyn-geal brushing and tissue samples from NPC patients as compared with those from healthy/nasopharyngitis indi-viduals.Four EBV-encoded microRNAs,mir-bart1-5p,mir-bart5,mir-bart6-5p,and mir-bart17-5p,were significantly up-regulated in both NPC brushing and tissue samples compared with those from healthy/nasopharyngitis controls(P<0.001).EBV DNA was significantly increased in both nasopharyngeal brushing samples(P<0.001)and tissue sam-ples(P<0.001)from NPC patients in comparison with those from healthy controls.Conclusions:Nasopharyngeal brushing can obtain sufficient tumoral materials for the analysis of viral nucleic acid,including EBV-encoded microRNAs and EBV DNA.For the detection of TSG expression,nasopharyngeal brushings was feasible but inferior to tissue samples.This study confirms nasopharyngeal brushing as an applicable sampling method that can aid in nucleic acid-based NPC research.
基金D.A.Megger and B.Sitek acknowledge the financial support from the PROFILE-consortium Ruhr.PROFILE is co-funded by the European Unionthe German federal state North Rhine-Westphaliaproject number z0911bt004e
文摘Proteomics has evolved into a powerful and widely used bioanalytical technique in the study of cancer,especially hepatocellular carcinoma (HCC).In this review,we provide an up to date overview of feasible proteome-analytical techniques for clinical questions.In addition,we present a broad summaryof proteornic studies of HCC utilizing various technical approaches for the analysis of samples derived from diverse sources like HCC cell lines,animal models,human tissue and body fluids.
