Oncological microdissection testicular sperm extraction(onco-micro-TESE)represents a significant breakthrough for patients with nonobstructive azoospermia(NOA)and a concomitant in situ testicular tumor,to be managed a...Oncological microdissection testicular sperm extraction(onco-micro-TESE)represents a significant breakthrough for patients with nonobstructive azoospermia(NOA)and a concomitant in situ testicular tumor,to be managed at the time of sperm retrieval.Onco-micro-TESE addresses the dual objectives of treating both infertility and the testicular tumor simultaneously.The technique is intricate,necessitating a comprehensive understanding of testicular anatomy,physiology,tumor biology,and advanced microsurgical methods.It aims to carefully extract viable spermatozoa while minimizing the risk of tumor dissemination.This review encapsulates the procedural intricacies,evaluates success determinants,including tumor pathology and spermatogenic tissue health,and discusses the implementation of imaging techniques for enhanced surgical precision.Ethical considerations are paramount,as the procedure implicates complex decision-making that weighs the potential oncological risks against the profound desire for fatherhood using the male gametes.The review aims to provide a holistic overview of onco-micro-TESE,detailing methodological advances,clinical outcomes,and the ethical landscape,thus offering an indispensable resource for clinicians navigating this multifaceted clinical scenario.展开更多
Dear Editor,Ciliary body masses are diagnostically challenging due to their hidden location,diverse pathologies,and limited examination methods[1].We report a case of a ciliary body inflammatory mass treated with tran...Dear Editor,Ciliary body masses are diagnostically challenging due to their hidden location,diverse pathologies,and limited examination methods[1].We report a case of a ciliary body inflammatory mass treated with trans-scleral excision and antiinflammatory therapy,preserving functional vision.展开更多
Objective Previous studies link lower body mass index(BMI)with increased obsessive-compulsive disorder(OCD)risk,yet other body mass indicators may be more etioloically relevant.We dissected the causal association betw...Objective Previous studies link lower body mass index(BMI)with increased obsessive-compulsive disorder(OCD)risk,yet other body mass indicators may be more etioloically relevant.We dissected the causal association between body fat mass(FM)and OCD.Methods Summary statistics from genome-wide association studies of European ancestry were utilized to conduct two-sample Mendelian randomization analysis.Heterogeneity,horizontal pleiotropy,and sensitivity analyses were performed to assess the robustness.Results The inverse variance weighting method demonstrated that a genetically predicted decrease in FM was causally associated with an increased OCD risk[odds ratio(OR)=0.680,95%confidence interval(CI):0.528–0.875,P=0.003].Similar estimates were obtained using the weighted median approach(OR=0.633,95%CI:0.438–0.915,P=0.015).Each standard deviation increases in genetically predicted body fat percentage corresponded to a reduced OCD risk(OR=0.638,95%CI:0.455–0.896,P=0.009).The sensitivity analysis confirmed the robustness of these findings with no outlier instrument variables identified.Conclusion The negative causal association between FM and the risk of OCD suggests that the prevention or treatment of mental disorders should include not only the control of BMI but also fat distribution and body composition.展开更多
Although traditional gamma-gamma density(GGD)logging technology is widely utilized,its potential environmental risks have prompted the development of more environmentally friendly neutron-gamma density(NGD)logging tec...Although traditional gamma-gamma density(GGD)logging technology is widely utilized,its potential environmental risks have prompted the development of more environmentally friendly neutron-gamma density(NGD)logging technology.However,NGD measurements are influenced by both neutron and gamma radiations.In the logging environment,variations in the formation composition indicate different elemental compositions,which affect the neutron-gamma reaction cross-sections and gamma generation.Compared to traditional gamma sources such as Cs-137,these changes significantly affect the generation and transport of neutron-induced inelastic gamma rays and hinder accurate measurements.To address this,a novel method is proposed that incorporates the mass attenuation coefficient function to account for the effects of various lithologies and pore contents on gamma-ray attenuation,thereby achieving more accurate density measurements by clarifying the transport processes of inelastic gamma rays with varying energies and spatial distributions in varied logging environments.The proposed method avoids the complex correction of neutron transport and is verified through Monte Carlo simulations for its applicability across various lithologies and pore contents,demonstrating absolute density errors that are less than 0.02 g/cm^(3)in clean formations and indicating good accuracy.This study clarifies the NGD mechanism and provides theoretical guidance for the application of NGD logging methods.Further studies will be conducted on extreme environmental conditions and tool calibration.展开更多
Although the effectiveness of a tuned viscous mass damper(TVMD)as an inerter-based device for vibration control in civil structures has been thoroughly investigated,there is a lack of systematic research regarding the...Although the effectiveness of a tuned viscous mass damper(TVMD)as an inerter-based device for vibration control in civil structures has been thoroughly investigated,there is a lack of systematic research regarding the application of TVMDs for seismic response control of industrial buildings coupled with mechanical equipment.Therefore,this study proposes ungrounded and grounded TVMDs to effectively utilize the mass of the mechanical equipment and fully exploit the capabilities of the inerter element.An optimal design methodology is developed by pursuing the maximum effective damping ratio and seeking the most rational TVMD control scheme.Validation of TVMD control performance is conducted through time-history analysis based on 20 real seismic ground motions recommended by ATC-40,and by providing a barrel mixer industrial building as a real-life numerical example.The results show that both an ungrounded and grounded TVMD can effectively mitigate the seismic response of the primary structure.Compared to the traditional tuned mass damper(TMD),TVMDs can obtain improved control performance for a given equipment mass ratio.Moreover,an ungrounded TVMD and a TMD show similar working mechanisms that tend to release the displacement of equipment to keep their optimal state,whereas equipment displacement for a grounded TVMD should be strictly limited to provide sufficient anti-force.展开更多
Mass wasting is globally recognized as a key geomorphic agent in permanent gully expansion and greatly contributes to watershed sediment losses.Though its formation process has been assessed by some physical models,th...Mass wasting is globally recognized as a key geomorphic agent in permanent gully expansion and greatly contributes to watershed sediment losses.Though its formation process has been assessed by some physical models,the occurrence and rainfall threshold have been rarely documented.In this study,rainfall-induced mass wasting events in two permanent gullies located in the Mollisols region of Northeast China,with Mollisols(gully 1)and sandy soil(gully 2)underneath were observed,and their differences were explored based on their soil strengths,hydraulic properties,excess topographies,and theoretical rainfall amounts.The sandy soil had a higher strength,faster pore water pressure dissipation rate,and lower suction stress at a specific soil moisture content compared to the black soil.The erosion thickness of the gully bed and sidewalls in gully 1 was shallower compared to gully 2.This was confirmed by the relationship between the erosion thickness and excess topography.The differences in the mass wasting erosion of the gully bed and sidewalls were due to the higher shear strength and well-drained hydraulic properties of the sandy soil compared to the black-soil.An infinite model was chosen to examine the temporal order of the mass wasting in the two gullies.It was found that the mass wasting in gully 2 occurred earlier than that in gully 1.This was likely due to the occurrence of an intense storm with less rainfall at the location of gully 2,while a light storm with heavier rainfall occurred in the location of gully 1.As Mollisols and sandy soil are the typical soil horizons in the Mollisols region worldwide,the results of this work could provide insightful knowledge for understanding the physical process of permanent gully expansion,which may be helpful for developing prediction models for sediment losses in some watersheds with vast Mollisols and highly developed gully system.展开更多
Pulsed dynamic electrolysis(PDE),driven by renewable energy,has emerged as an innovative electrocatalytic conversion method,demonstrating significant potential in addressing global energy challenges and promoting sust...Pulsed dynamic electrolysis(PDE),driven by renewable energy,has emerged as an innovative electrocatalytic conversion method,demonstrating significant potential in addressing global energy challenges and promoting sustainable development.Despite significant progress in various electrochemical systems,the regulatory mechanisms of PDE in energy and mass transfer and the lifespan extension of electrolysis systems,particularly in water electrolysis(WE)for hydrogen production,remain insufficiently explored.Therefore,there is an urgent need for a deeper understanding of the unique contributions of PDE in mass transfer enhancement,microenvironment regulation,and hydrogen production optimization,aiming to achieve low-energy consumption,high catalytic activity,and long-term stability in the generation of target products.Here,this review critically examines the microenvironmental effects of PDE on energy and mass transfer,the electrode degradation mechanisms in the lifespan extension of electrolysis systems,and the key factors in enhancing WE for hydrogen production,providing a comprehensive summary of current research progress.The review focuses on the complex regulatory mechanisms of frequency,duty cycle,amplitude,and other factors in hydrogen evolution reaction(HER)performance within PDE strategies,revealing the interrelationships among them.Finally,the potential future directions and challenges for transitioning from laboratory studies to industrial applications are proposed.展开更多
This study examined non-uniform loading in goaf cantilever rock masses via testing,modeling,and mechanical analysis to solve instantaneous fracture and section buckling from mining abutment pressure.The study investig...This study examined non-uniform loading in goaf cantilever rock masses via testing,modeling,and mechanical analysis to solve instantaneous fracture and section buckling from mining abutment pressure.The study investigates the non-uniform load gradient effect on fracture characteristics,including load characteristics,fracture location,fracture distribution,and section roughness.A digital model for fracture interface buckling analysis was developed,elucidating the influence of non-uniform load gradients on Fracture Interface Curvature(FIC),Buckling Rate of Change(BRC),and Buckling Domain Field(BDF).The findings reveal that nonlinear tensile stress concentration and abrupt tensile-compressive-shear strain mutations under non-uniform loading are fundamental mechanisms driving fracture path buckling in cantilever rock mass structures.The buckling process of rock mass under non-uniform load can be divided into two stages:low load gradient and high gradient load.In the stage of low gradient load,the buckling behavior is mainly reflected in the compression-shear fracture of the edge.In the stage of high gradient load,a buckling band along the loading direction is gradually formed in the rock mass.These buckling principles establish a theoretical basis for accurately characterizing bearing fractures,fracture interface instability,and vibration sources within overlying cantilever rock masses in goaf.展开更多
The Three Gorges Reservoir area hosts numerous high-risk columnar rock masses,but predicting their full-process of collapse dynamics in narrow-deep terrains remains challenging.This study investigated the Changdongzi ...The Three Gorges Reservoir area hosts numerous high-risk columnar rock masses,but predicting their full-process of collapse dynamics in narrow-deep terrains remains challenging.This study investigated the Changdongzi columnar dangerous rock mass(CCDRM)through field surveys and simulated its collapse dynamics with Material Point Method(MPM),and analyzed the collapse-diffusionsurge generation mechanisms.Field investigations revealed a 0.8 m-thick lotus-like extrusion structure at the base,severely degraded by cyclic water fluctuations,predisposing the rock mass to base-crushing failure.Numerical simulations indicate collapse velocities up to 42 m/s,amplified spreading with a 318 m deposition range and 11 m maximum thickness,and two major collisions causing 50%-60%velocity reduction.Gully topography buffers kinetic energy,retaining 5%debris on slopes and channeling 7%into the river.High-speed river impact generates a 20 m-diameter cavity,producing an 11 m surge,identifying the Guandukou Town waterfront 1.2 km downstream as a high-risk zone.These results clarify the collapse mechanisms of columnar dangerous rock mass and inform its hazard mitigation strategies.展开更多
Background:Diabetic foot,a severe complication of diabetes,is characterized by chronic refractory wounds.Sanhuang Oil,a topical herbal formula,demonstrates significant therapeutic effects including antibacterial,anti-...Background:Diabetic foot,a severe complication of diabetes,is characterized by chronic refractory wounds.Sanhuang Oil,a topical herbal formula,demonstrates significant therapeutic effects including antibacterial,anti-inflammatory,and immunomodulatory activities.However,its active constituents and mechanisms of action against diabetic foot remain to be elucidated.Methods:In this study,the chemical constituents of Sanhuang Oil were identified using UPLC-QE-Orbitrap-MS.Subsequently,the mechanism by which Sanhuang Oil promotes diabetic foot ulcer healing was predicted by integrating network pharmacology and molecular docking.Additionally,diabetic mouse model was established in ICR mice using a combination of a high-fat diet(HFD)and streptozotocin(STZ)chemical induction.A full-thickness skin defect was created on the dorsum of the mice.Wound healing and the healing rate were observed following Sanhuang Oil intervention.The mechanism underlying Sanhuang Oil’s promotion of diabetic ulcer healing was further investigated using transcriptomics and histopathological examination(H&E staining).Results:A total of 97 active ingredients were identified from Sanhuang Oil.Network pharmacology analysis predicted 543 common targets,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis identified 203 relevant pathways.Molecular docking further confirmed high binding affinity(binding energy≤−5.0 kcal/mol)between specific active components in Sanhuang Oil(e.g.,coptisine,phellodendrine,baicalein)and key targets associated with diabetic foot ulcers(e.g.,EGFR,AKT1,STAT3).In vivo experimental results demonstrated that the wound healing rate was significantly higher in Sanhuang Oil-treated groups compared to the model group(P<0.001).HE staining revealed that the high-dose Sanhuang Oil group exhibited more pronounced epithelial tissue coverage over the wound,reduced inflammatory cell infiltration,and increased collagen deposition and fibroblast proliferation.transcriptomic analysis identified Pdk4,Ttn,Csrp3,Actn2,Myoz2,Tnnc2,Myod1,Myog,Myot,and Myf6 as key regulatory proteins involved in promoting wound healing.Conclusion:Sanhuang Oil promotes wound healing in diabetic ulcer mice,potentially by mitigating inflammation and regulating key targets such as Pdk4 to enhance fibroblast function.These findings provide novel insights into the multi-target,multi-pathway mechanism of Sanhuang Oil for treating diabetic foot ulcers.展开更多
Overweight and obesity has been a major public health problem globally.It was estimated that more than 2.1 billion adults were affected by overweight or obese in 2021 worldwide,about one fifth of whom lived in China^(...Overweight and obesity has been a major public health problem globally.It was estimated that more than 2.1 billion adults were affected by overweight or obese in 2021 worldwide,about one fifth of whom lived in China^([1]).By 2050,the country is forecast to remain the one with the largest population of overweight and obese globally^([1]),if no effective strategies were applied on overweight/obesity control.展开更多
To investigate the long-term stability of soft-hard interbedded rock masses with initial damage induced by earthquakes and periodic drying and wetting,this study prepared samples with different initial damage through ...To investigate the long-term stability of soft-hard interbedded rock masses with initial damage induced by earthquakes and periodic drying and wetting,this study prepared samples with different initial damage through cyclic loading and unloading(CLU)experiments followed by cyclic drying and wetting(CDW)experiments,and finally conducted creep experiments.The study analyzed the effects of initial damage on creep mechanical behavior,crack evolution,and explored failure precursor information,revealing the damage failure mechanisms.The results show that the structural characteristics of the rock mass control its macroscopic failure mode.Initial damage promotes microcrack development,influences the fracture mode,and increases the proportion of high-frequency(200−280 kHz)acoustic emission events during creep.Meanwhile,initial damage exacerbates creep characteristics,increasing the creep rate,shortening total creep failure time,and reducing long-term strength.The damage failure is attributed to:the generation of internal cracks and pores in the rock caused by CLU;mineral hydrolysis and expansion-contraction due to CDW,resulting in weakened intergranular cementation;and full development of cracks and pores under creep stress.Additionally,the deformation difference coefficient and the coefficient of variation of RA/AF values can serve as precursor indicators for creep failure.展开更多
The neuroinflammatory response mediated by microglial activation plays an important role in the secondary nerve injury of traumatic brain injury.The post-transcriptional modification of N^(6)-methyladenosine is ubiqui...The neuroinflammatory response mediated by microglial activation plays an important role in the secondary nerve injury of traumatic brain injury.The post-transcriptional modification of N^(6)-methyladenosine is ubiquitous in the immune response of the central nervous system.The fat mass and obesity-related protein catalyzes the demethylation of N^(6)-methyladenosine modifications on mRNA and is widely expressed in various tissues,participating in the regulation of multiple diseases’biological processes.However,the role of fat mass and obesity in microglial activation and the subsequent neuroinflammatory response after traumatic brain injury is unclear.In this study,we found that the expression of fat mass and obesity was significantly down-regulated in both lipopolysaccharide-treated BV2 cells and a traumatic brain injury mouse model.After fat mass and obesity interference,BV2 cells exhibited a pro-inflammatory phenotype as shown by the increased proportion of CD11b^(+)/CD86^(+)cells and the secretion of pro-inflammatory cytokines.Fat mass and obesity-mediated N^(6)-methyladenosine demethylation accelerated the degradation of ADAM17 mRNA,while silencing of fat mass and obesity enhanced the stability of ADAM17 mRNA.Therefore,down-regulation of fat mass and obesity expression leads to the abnormally high expression of ADAM17 in microglia.These results indicate that the activation of microglia and neuroinflammatory response regulated by fat mass and obesity-related N^(6)-methyladenosine modification plays an important role in the pro-inflammatory process of secondary injury following traumatic brain injury.展开更多
The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6V...The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6VS was microdissected with a needle and transferred into a 0.5 mL Ep tube. In the 'single tube', all the subsequence steps were conducted. After two round of LA (Linker adaptor)_PCR amplification, the size of PCR bands ranged from 100 to 3 000 bp, with predominate bands 600-1 500 bp. The products were confirmed by Southern blotting analysis using Haynaldia villosa (L.) Schur. genomic DNA labeled with 32 P as probe. The PCR products were purified and ligated into clone vector-pGEM_T easy vector. Then, the plasmids were transformed into competence E. coli JM109 with cool CaCl 2. It was estimated that there were more than 17 000 white clones in the library. The size of insert fragments distributed from 100-1 500 bp, with average of 600 bp. Using H. villosa genomic DNA as probe, dot blotting results showed that 37% clones displayed strong and medium positive signals, and 63% clones had faint or no signals. It is demonstrated that there were about 37% repeat sequence clones and 67% single/unique sequence clones in the library. Eight H. villosa_specific clones were screened from the library, and two clones pHVMK22 and pHVMK134 were used for RFLP analysis and sequencing. Both of them were H. villosa specific clones. The pHVMK22 was a unique sequence clone, and the pHVMK134 was a repeat sequence clone. When the pHVMK22 was used as a probe for Southern hybridization, all the powdery mildew resistance materials showed a special band of 2 kb, while all the susceptible ones not. The pHVMK22 may be applied to detect the existence of Pm21.展开更多
Objective: Laser capture microdisection has become indispensable to the analysis of the difference of gene expression between human bladder transitional cell and bladder transitional cell carcinoma (BTCC). However,...Objective: Laser capture microdisection has become indispensable to the analysis of the difference of gene expression between human bladder transitional cell and bladder transitional cell carcinoma (BTCC). However, to obtain sufficient RNA from laser-capture microdissected cells is quite difficult. The study was designed to determinc a feasible technical routine to isolate transitional cells from bladder membrane, separate carcinoma cclls from stromal cells and to amplify the RNA isolated from laser-capture microdissected cells. Methods: Bladder transitional cell were obtained from frozen sections of bladder membrane applying LCM, by the same token, BTCC cells from frozen sections of BTCC tissue. Then RNA was extracted and linearly amplified in vitro. The expression levels of β-actin in primary total RNA and amplified RNA were detected using RT-PCR. Results: That RNA integrity was good after LCM was confirmed by control experiment Ⅰ; By control experiment Ⅱ, the correlation between the number of LCM-shooting and RNA quantity undcr arranged conditions was preliminarily confirmed. About 0.5-2.5kb RNA fragments were obtained after RNA amplification and β-actin levels were integral. Conclusion: Laser capture microdissection combined with RNA linear amplification in vitro can be successfully applied to obtain pure objective cells for research. The integrity of the amplified RNA is good and can be employed in further research.展开更多
We performed this meta-analysis to evaluate the predictive value of different parameters in the sperm retrieval rate (SRR) of microdissection testicular sperm extraction (TESE) in patients with nonobstructive azoo...We performed this meta-analysis to evaluate the predictive value of different parameters in the sperm retrieval rate (SRR) of microdissection testicular sperm extraction (TESE) in patients with nonobstructive azoospermia (NOA). All relevant studies were searched in PubMed, Web of Science, EMBASE, Cochrane Library, and EBSCO. We chose three parameters to perform the meta-analysis: follicle-stimulating hormone (FSH), testicular volume, and testicular histopathological findings which included three patterns: hypospermatogenesis (HS), maturation arrest (MA), and Sertoli-cell-only syndrome (SCOS). If there was a threshold effect, only the area under the summary receiver operating characteristic curve (AUSROC) was calculated. Otherwise, the pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), and the diagnostic odds ratio (DOR) were also calculated. Twenty-one articles were included in our study finally. There was a threshold effect among studies investigating FSH and SCOS. The AUSROCs of FSH, testicular volume, HS, MA, and SCOS were 0.6119, 0.6389, 0.6758, 0.5535, and 0.2763, respectively. The DORs of testicular volume, HS, and MA were 1.98, 16.49, and 1.26, respectively. The sensitivities of them were 0.80, 0.30, and 0.27, while the specificities of them were 0.35, 0.98, and 0.76, respectively. The PLRs of them were 1.49, 10.63, and 1.15, respectively. And NLRs were 0.73, 0.72, and 0.95, respectively. All the investigated factors in our study had limited predictive value. However, the histopathological findings were helpful to some extent. Most patients with HS could get sperm by microdissection TESE.展开更多
In 220 consecutive patients with non-obstructive azoospermia, sperm retrieval was attempted by a combination of conventional and microdissection testicular sperm extraction (TESE). For sperm retrieval, 2-3 conventio...In 220 consecutive patients with non-obstructive azoospermia, sperm retrieval was attempted by a combination of conventional and microdissection testicular sperm extraction (TESE). For sperm retrieval, 2-3 conventional biopsies were performed followed by a microdissection TESE in cases of negative conventional biopsies. During the surgery, the vasculature of the testis was assessed using the operative microscope, and the location of positive biopsies was registered in relation to the blood supply. The overall sperm retrieval rate was 58.2%. From the initial conventional biopsies, sperm could be retrieved in 46.8% of the patients. With microdissection TESE, sperm could be retrieved from an additional 11.4% of the patients. The further use of microdissection TESE improved the sperm retrieval rate significantly (P=0.017). No significant accumulation of positive biopsies was found towards the rete testis or the main testicular vessels.展开更多
AIM. To study the differential gene expression profiles of target cells in primary gastric cancer and its metastatic lymph nodes using laser microdissection (LMD) in combination with cDNA microarray. METHODS: Norma...AIM. To study the differential gene expression profiles of target cells in primary gastric cancer and its metastatic lymph nodes using laser microdissection (LMD) in combination with cDNA microarray. METHODS: Normal gastric tissue samples from 30 healthy individuals, 36 cancer tissue samples from primary gastric carcinoma and lymph node metastasis tissue samples from 58 patients during gastric cancer resection were obtained using LMD in combination with cDNA microarray independently. After P27-based amplification, aRNA from 36 of 58 patients (group 1) with lymph node metastasis and metastatic tissue specimens from the remaining 22 patients (group 2) were applied to cDNA microarray. Semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) and imrnunohistochemical assay verified the results of microarray in group 2 and further identified genes differentially expressed in the progression of gastric cancer. RESULTS: The expression of 10 genes was up-regulated while the expression of 15 genes was down-regulated in 22 gastric carcinoma samples compared with that of genes in the normal controls. The results were confirmed at the level of mRNA and protein, and suggested that four genes (OPCML, RNASE1, YES1 and ACK1) could play a key role in the tumorigenesis and metastasis of gastric cancer. The expression pattern of 3 genes (OPCML, RNASE1 and YES1) was similar to tumor suppressor genes. For example, the expression level of these genes was the highest in normal gastric epithelium, which was decreased in primary carcinoma, and further decreased in metastatic lymph nodes. On the contrary, the expression pattern of gene ACK1 was similar to that of oncogene. Four genes were further identified as differentially expressed genes in the majority of the cases in the progression of gastric cancer. CONCLUSION: LMD in combination with cDNA microaro ray provides a unique support foe the identification of early expression profiles of differential genes and the expression pattern of 3 genes (OPCML, RNASE1 and YES1) associated with the progression of gastric cancer. Further study is needed to reveal the molecular mechanism of lymph node metastasis in patients with gastric cancer.展开更多
AIM: To examine the gene expression profile of gastric cancer (GC) by combination of laser capture microdissection (LCM) and microarray and to correlate the profiling with histological subtypes. METHODS: Using L...AIM: To examine the gene expression profile of gastric cancer (GC) by combination of laser capture microdissection (LCM) and microarray and to correlate the profiling with histological subtypes. METHODS: Using LCM, pure cancer cells were procured from 45 cancerous tissues. After procurement of about 5 000 cells, total RNA was extracted and the quality of RNA was determined before further amplification and hybridization. One microgram of amplified RNA was converted to cDNA and hybridized to cDNA microarray. RESULTS: Among 45 cases, only 21 were qualified for their RNAs. A total of 62 arrays were performed. These included 42 arrays for cancer (21 cases with dyeswab duplication) and 20 arrays for non-tumorous cells (10 cases with dye-swab duplication) with universal reference. Analyzed data showed 504 genes were differentially expressed and could distinguish cancerous and non-cancerous groups with more than 99% accuracy. Of the 504 genes, trefoil factors 1, 2, and 3 were in the list and their expression patterns were consistent with previous reports. Immunohistochemical staining of trefoil factor 1 was also consistent with the array data. Analyses of the tumor group with these 504 genes showed that there were 3 subgroups of GC that did not correspond to any current classification system, including Lauren's dassification. CONCLUSION: By using LCM, linear amplification of RNA, and cDNA microarray, we have identified a panel of genes that have the power to discriminate between GC and non-cancer groups. The new molecular classification and the identified novel genes in gastric carcinogenesis deserve further investigations to elucidate their clinicopathological significance.展开更多
基金supported by the National Natural Science Foundation of China(No.82371633)Peking University Clinical Scientist Training Program and the Fundamental Research Funds for the Central University(BMU2023PYJ H012).
文摘Oncological microdissection testicular sperm extraction(onco-micro-TESE)represents a significant breakthrough for patients with nonobstructive azoospermia(NOA)and a concomitant in situ testicular tumor,to be managed at the time of sperm retrieval.Onco-micro-TESE addresses the dual objectives of treating both infertility and the testicular tumor simultaneously.The technique is intricate,necessitating a comprehensive understanding of testicular anatomy,physiology,tumor biology,and advanced microsurgical methods.It aims to carefully extract viable spermatozoa while minimizing the risk of tumor dissemination.This review encapsulates the procedural intricacies,evaluates success determinants,including tumor pathology and spermatogenic tissue health,and discusses the implementation of imaging techniques for enhanced surgical precision.Ethical considerations are paramount,as the procedure implicates complex decision-making that weighs the potential oncological risks against the profound desire for fatherhood using the male gametes.The review aims to provide a holistic overview of onco-micro-TESE,detailing methodological advances,clinical outcomes,and the ethical landscape,thus offering an indispensable resource for clinicians navigating this multifaceted clinical scenario.
基金Supported by National Natural Science Foundation of China(No.82101113).
文摘Dear Editor,Ciliary body masses are diagnostically challenging due to their hidden location,diverse pathologies,and limited examination methods[1].We report a case of a ciliary body inflammatory mass treated with trans-scleral excision and antiinflammatory therapy,preserving functional vision.
基金supported by the Yanzhao Gold Talent Project of Hebei Province(NO.HJZD202506)。
文摘Objective Previous studies link lower body mass index(BMI)with increased obsessive-compulsive disorder(OCD)risk,yet other body mass indicators may be more etioloically relevant.We dissected the causal association between body fat mass(FM)and OCD.Methods Summary statistics from genome-wide association studies of European ancestry were utilized to conduct two-sample Mendelian randomization analysis.Heterogeneity,horizontal pleiotropy,and sensitivity analyses were performed to assess the robustness.Results The inverse variance weighting method demonstrated that a genetically predicted decrease in FM was causally associated with an increased OCD risk[odds ratio(OR)=0.680,95%confidence interval(CI):0.528–0.875,P=0.003].Similar estimates were obtained using the weighted median approach(OR=0.633,95%CI:0.438–0.915,P=0.015).Each standard deviation increases in genetically predicted body fat percentage corresponded to a reduced OCD risk(OR=0.638,95%CI:0.455–0.896,P=0.009).The sensitivity analysis confirmed the robustness of these findings with no outlier instrument variables identified.Conclusion The negative causal association between FM and the risk of OCD suggests that the prevention or treatment of mental disorders should include not only the control of BMI but also fat distribution and body composition.
基金supported by the National Natural Science Foundation of China(U23B20151 and 52171253).
文摘Although traditional gamma-gamma density(GGD)logging technology is widely utilized,its potential environmental risks have prompted the development of more environmentally friendly neutron-gamma density(NGD)logging technology.However,NGD measurements are influenced by both neutron and gamma radiations.In the logging environment,variations in the formation composition indicate different elemental compositions,which affect the neutron-gamma reaction cross-sections and gamma generation.Compared to traditional gamma sources such as Cs-137,these changes significantly affect the generation and transport of neutron-induced inelastic gamma rays and hinder accurate measurements.To address this,a novel method is proposed that incorporates the mass attenuation coefficient function to account for the effects of various lithologies and pore contents on gamma-ray attenuation,thereby achieving more accurate density measurements by clarifying the transport processes of inelastic gamma rays with varying energies and spatial distributions in varied logging environments.The proposed method avoids the complex correction of neutron transport and is verified through Monte Carlo simulations for its applicability across various lithologies and pore contents,demonstrating absolute density errors that are less than 0.02 g/cm^(3)in clean formations and indicating good accuracy.This study clarifies the NGD mechanism and provides theoretical guidance for the application of NGD logging methods.Further studies will be conducted on extreme environmental conditions and tool calibration.
基金National Natural Science Foundation of China under Grant Nos.52408327 and 52278306Key Research and Development Program of Hunan Province,China under Grant No.2022SK2096+3 种基金Science and Technology Progress and Innovation Project of the Department of Transportation of Hunan Province,China under Grant No.201912Natural Science Foundation of Hunan Province,China under Grant No.2024JJ6198Scientific Research Project of the Education Department of Hunan Province,China under Grant No.25A0645Emergency Management Science and Technology Project of the Emergency Management Department of Hunan Province,China under Grant No.yjtkjxm_202406。
文摘Although the effectiveness of a tuned viscous mass damper(TVMD)as an inerter-based device for vibration control in civil structures has been thoroughly investigated,there is a lack of systematic research regarding the application of TVMDs for seismic response control of industrial buildings coupled with mechanical equipment.Therefore,this study proposes ungrounded and grounded TVMDs to effectively utilize the mass of the mechanical equipment and fully exploit the capabilities of the inerter element.An optimal design methodology is developed by pursuing the maximum effective damping ratio and seeking the most rational TVMD control scheme.Validation of TVMD control performance is conducted through time-history analysis based on 20 real seismic ground motions recommended by ATC-40,and by providing a barrel mixer industrial building as a real-life numerical example.The results show that both an ungrounded and grounded TVMD can effectively mitigate the seismic response of the primary structure.Compared to the traditional tuned mass damper(TMD),TVMDs can obtain improved control performance for a given equipment mass ratio.Moreover,an ungrounded TVMD and a TMD show similar working mechanisms that tend to release the displacement of equipment to keep their optimal state,whereas equipment displacement for a grounded TVMD should be strictly limited to provide sufficient anti-force.
基金primarily supported by the National Key Research and Development Program of China(Grant No.2024YFD1501201-3)partially supported by the National Key Research and Development Program of China(Grant No.2021YFD1500700)。
文摘Mass wasting is globally recognized as a key geomorphic agent in permanent gully expansion and greatly contributes to watershed sediment losses.Though its formation process has been assessed by some physical models,the occurrence and rainfall threshold have been rarely documented.In this study,rainfall-induced mass wasting events in two permanent gullies located in the Mollisols region of Northeast China,with Mollisols(gully 1)and sandy soil(gully 2)underneath were observed,and their differences were explored based on their soil strengths,hydraulic properties,excess topographies,and theoretical rainfall amounts.The sandy soil had a higher strength,faster pore water pressure dissipation rate,and lower suction stress at a specific soil moisture content compared to the black soil.The erosion thickness of the gully bed and sidewalls in gully 1 was shallower compared to gully 2.This was confirmed by the relationship between the erosion thickness and excess topography.The differences in the mass wasting erosion of the gully bed and sidewalls were due to the higher shear strength and well-drained hydraulic properties of the sandy soil compared to the black-soil.An infinite model was chosen to examine the temporal order of the mass wasting in the two gullies.It was found that the mass wasting in gully 2 occurred earlier than that in gully 1.This was likely due to the occurrence of an intense storm with less rainfall at the location of gully 2,while a light storm with heavier rainfall occurred in the location of gully 1.As Mollisols and sandy soil are the typical soil horizons in the Mollisols region worldwide,the results of this work could provide insightful knowledge for understanding the physical process of permanent gully expansion,which may be helpful for developing prediction models for sediment losses in some watersheds with vast Mollisols and highly developed gully system.
基金financially supported by the Key Research and Development Program of Heilongjiang Province(No.2024ZXJ03C06)National Natural Science Foundation of China(No.52476192,No.52106237)+1 种基金Natural Science Foundation of Heilongjiang Province(No.YQ2022E027)Technology Project of China Datang Technology Innovation Co.,Ltd(No.DTKC-2024-20610).
文摘Pulsed dynamic electrolysis(PDE),driven by renewable energy,has emerged as an innovative electrocatalytic conversion method,demonstrating significant potential in addressing global energy challenges and promoting sustainable development.Despite significant progress in various electrochemical systems,the regulatory mechanisms of PDE in energy and mass transfer and the lifespan extension of electrolysis systems,particularly in water electrolysis(WE)for hydrogen production,remain insufficiently explored.Therefore,there is an urgent need for a deeper understanding of the unique contributions of PDE in mass transfer enhancement,microenvironment regulation,and hydrogen production optimization,aiming to achieve low-energy consumption,high catalytic activity,and long-term stability in the generation of target products.Here,this review critically examines the microenvironmental effects of PDE on energy and mass transfer,the electrode degradation mechanisms in the lifespan extension of electrolysis systems,and the key factors in enhancing WE for hydrogen production,providing a comprehensive summary of current research progress.The review focuses on the complex regulatory mechanisms of frequency,duty cycle,amplitude,and other factors in hydrogen evolution reaction(HER)performance within PDE strategies,revealing the interrelationships among them.Finally,the potential future directions and challenges for transitioning from laboratory studies to industrial applications are proposed.
基金support provided by the National Natural Science Foundation of China(No.52274077)the Natural Science Foundation of Henan(No.242300421072)+2 种基金the Youth Elite Teachers Cultivation Program for Higher Education Institutions in Henan Province(No.2024GGJS036)the Funds for Distinguished Young Scholars of Henan Polytechnic University(No.J2023-3)the Young Core Teacher Funding Scheme of Henan Polytechnic University(No.2023XQG-09).
文摘This study examined non-uniform loading in goaf cantilever rock masses via testing,modeling,and mechanical analysis to solve instantaneous fracture and section buckling from mining abutment pressure.The study investigates the non-uniform load gradient effect on fracture characteristics,including load characteristics,fracture location,fracture distribution,and section roughness.A digital model for fracture interface buckling analysis was developed,elucidating the influence of non-uniform load gradients on Fracture Interface Curvature(FIC),Buckling Rate of Change(BRC),and Buckling Domain Field(BDF).The findings reveal that nonlinear tensile stress concentration and abrupt tensile-compressive-shear strain mutations under non-uniform loading are fundamental mechanisms driving fracture path buckling in cantilever rock mass structures.The buckling process of rock mass under non-uniform load can be divided into two stages:low load gradient and high gradient load.In the stage of low gradient load,the buckling behavior is mainly reflected in the compression-shear fracture of the edge.In the stage of high gradient load,a buckling band along the loading direction is gradually formed in the rock mass.These buckling principles establish a theoretical basis for accurately characterizing bearing fractures,fracture interface instability,and vibration sources within overlying cantilever rock masses in goaf.
基金supported by the Regional Innovation Development Joint Fund of the National Natural Science Foundation of China(U23A2045)the scientific research project of the Chongqing Municipal Bureau of Planning and Natural Resources(KJ-2023046)the scientific and technological project of the Hubei Provincial Bureau of Geological Survey(KJ2024-34)。
文摘The Three Gorges Reservoir area hosts numerous high-risk columnar rock masses,but predicting their full-process of collapse dynamics in narrow-deep terrains remains challenging.This study investigated the Changdongzi columnar dangerous rock mass(CCDRM)through field surveys and simulated its collapse dynamics with Material Point Method(MPM),and analyzed the collapse-diffusionsurge generation mechanisms.Field investigations revealed a 0.8 m-thick lotus-like extrusion structure at the base,severely degraded by cyclic water fluctuations,predisposing the rock mass to base-crushing failure.Numerical simulations indicate collapse velocities up to 42 m/s,amplified spreading with a 318 m deposition range and 11 m maximum thickness,and two major collisions causing 50%-60%velocity reduction.Gully topography buffers kinetic energy,retaining 5%debris on slopes and channeling 7%into the river.High-speed river impact generates a 20 m-diameter cavity,producing an 11 m surge,identifying the Guandukou Town waterfront 1.2 km downstream as a high-risk zone.These results clarify the collapse mechanisms of columnar dangerous rock mass and inform its hazard mitigation strategies.
基金supported by the Natural Science Foundation of Hubei Provincial Department of Education(D20232101)Shandong Second Medical University 2024 Affiliated Hospital(Teaching Hospital)Scientific Research Development Fund Project(2024FYQ026)+3 种基金the innovative Research Programme of Xiangyang No.1 People’s Hospital(XYY2023ZY01)Faculty Development Grants of Xiangyang No.1 People’s Hospital Affiliated to Hubei University of Medicine(XYY2023D05)Joint supported by Hubei Provincial Natural Science Foundation and Xiangyang of China(2025AFD091)Traditional Chinese Medicine Scientific Research Project of Hubei Provincial Administration of Traditional Chinese Medicine(ZY2025D019).
文摘Background:Diabetic foot,a severe complication of diabetes,is characterized by chronic refractory wounds.Sanhuang Oil,a topical herbal formula,demonstrates significant therapeutic effects including antibacterial,anti-inflammatory,and immunomodulatory activities.However,its active constituents and mechanisms of action against diabetic foot remain to be elucidated.Methods:In this study,the chemical constituents of Sanhuang Oil were identified using UPLC-QE-Orbitrap-MS.Subsequently,the mechanism by which Sanhuang Oil promotes diabetic foot ulcer healing was predicted by integrating network pharmacology and molecular docking.Additionally,diabetic mouse model was established in ICR mice using a combination of a high-fat diet(HFD)and streptozotocin(STZ)chemical induction.A full-thickness skin defect was created on the dorsum of the mice.Wound healing and the healing rate were observed following Sanhuang Oil intervention.The mechanism underlying Sanhuang Oil’s promotion of diabetic ulcer healing was further investigated using transcriptomics and histopathological examination(H&E staining).Results:A total of 97 active ingredients were identified from Sanhuang Oil.Network pharmacology analysis predicted 543 common targets,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis identified 203 relevant pathways.Molecular docking further confirmed high binding affinity(binding energy≤−5.0 kcal/mol)between specific active components in Sanhuang Oil(e.g.,coptisine,phellodendrine,baicalein)and key targets associated with diabetic foot ulcers(e.g.,EGFR,AKT1,STAT3).In vivo experimental results demonstrated that the wound healing rate was significantly higher in Sanhuang Oil-treated groups compared to the model group(P<0.001).HE staining revealed that the high-dose Sanhuang Oil group exhibited more pronounced epithelial tissue coverage over the wound,reduced inflammatory cell infiltration,and increased collagen deposition and fibroblast proliferation.transcriptomic analysis identified Pdk4,Ttn,Csrp3,Actn2,Myoz2,Tnnc2,Myod1,Myog,Myot,and Myf6 as key regulatory proteins involved in promoting wound healing.Conclusion:Sanhuang Oil promotes wound healing in diabetic ulcer mice,potentially by mitigating inflammation and regulating key targets such as Pdk4 to enhance fibroblast function.These findings provide novel insights into the multi-target,multi-pathway mechanism of Sanhuang Oil for treating diabetic foot ulcers.
文摘Overweight and obesity has been a major public health problem globally.It was estimated that more than 2.1 billion adults were affected by overweight or obese in 2021 worldwide,about one fifth of whom lived in China^([1]).By 2050,the country is forecast to remain the one with the largest population of overweight and obese globally^([1]),if no effective strategies were applied on overweight/obesity control.
基金Project(U22A20603)supported by the National Natural Science Foundation of ChinaProject(2023YFC3008300)supported by the National Key Research and Development Program of China。
文摘To investigate the long-term stability of soft-hard interbedded rock masses with initial damage induced by earthquakes and periodic drying and wetting,this study prepared samples with different initial damage through cyclic loading and unloading(CLU)experiments followed by cyclic drying and wetting(CDW)experiments,and finally conducted creep experiments.The study analyzed the effects of initial damage on creep mechanical behavior,crack evolution,and explored failure precursor information,revealing the damage failure mechanisms.The results show that the structural characteristics of the rock mass control its macroscopic failure mode.Initial damage promotes microcrack development,influences the fracture mode,and increases the proportion of high-frequency(200−280 kHz)acoustic emission events during creep.Meanwhile,initial damage exacerbates creep characteristics,increasing the creep rate,shortening total creep failure time,and reducing long-term strength.The damage failure is attributed to:the generation of internal cracks and pores in the rock caused by CLU;mineral hydrolysis and expansion-contraction due to CDW,resulting in weakened intergranular cementation;and full development of cracks and pores under creep stress.Additionally,the deformation difference coefficient and the coefficient of variation of RA/AF values can serve as precursor indicators for creep failure.
基金supported by grants from the Major Projects of Health Science Research Foundation for Middle-Aged and Young Scientist of Fujian Province,China,No.2022ZQNZD01010010the National Natural Science Foundation of China,No.82371390Fujian Province Scientific Foundation,No.2023J01725(all to XC).
文摘The neuroinflammatory response mediated by microglial activation plays an important role in the secondary nerve injury of traumatic brain injury.The post-transcriptional modification of N^(6)-methyladenosine is ubiquitous in the immune response of the central nervous system.The fat mass and obesity-related protein catalyzes the demethylation of N^(6)-methyladenosine modifications on mRNA and is widely expressed in various tissues,participating in the regulation of multiple diseases’biological processes.However,the role of fat mass and obesity in microglial activation and the subsequent neuroinflammatory response after traumatic brain injury is unclear.In this study,we found that the expression of fat mass and obesity was significantly down-regulated in both lipopolysaccharide-treated BV2 cells and a traumatic brain injury mouse model.After fat mass and obesity interference,BV2 cells exhibited a pro-inflammatory phenotype as shown by the increased proportion of CD11b^(+)/CD86^(+)cells and the secretion of pro-inflammatory cytokines.Fat mass and obesity-mediated N^(6)-methyladenosine demethylation accelerated the degradation of ADAM17 mRNA,while silencing of fat mass and obesity enhanced the stability of ADAM17 mRNA.Therefore,down-regulation of fat mass and obesity expression leads to the abnormally high expression of ADAM17 in microglia.These results indicate that the activation of microglia and neuroinflammatory response regulated by fat mass and obesity-related N^(6)-methyladenosine modification plays an important role in the pro-inflammatory process of secondary injury following traumatic brain injury.
基金国家"8 6 3"计划资助项目 (Z 17 0 4 0 1) 国家转基因植物研究与产业化资助项目 (J0 0 A 0 0 2 )~~
文摘The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6VS was microdissected with a needle and transferred into a 0.5 mL Ep tube. In the 'single tube', all the subsequence steps were conducted. After two round of LA (Linker adaptor)_PCR amplification, the size of PCR bands ranged from 100 to 3 000 bp, with predominate bands 600-1 500 bp. The products were confirmed by Southern blotting analysis using Haynaldia villosa (L.) Schur. genomic DNA labeled with 32 P as probe. The PCR products were purified and ligated into clone vector-pGEM_T easy vector. Then, the plasmids were transformed into competence E. coli JM109 with cool CaCl 2. It was estimated that there were more than 17 000 white clones in the library. The size of insert fragments distributed from 100-1 500 bp, with average of 600 bp. Using H. villosa genomic DNA as probe, dot blotting results showed that 37% clones displayed strong and medium positive signals, and 63% clones had faint or no signals. It is demonstrated that there were about 37% repeat sequence clones and 67% single/unique sequence clones in the library. Eight H. villosa_specific clones were screened from the library, and two clones pHVMK22 and pHVMK134 were used for RFLP analysis and sequencing. Both of them were H. villosa specific clones. The pHVMK22 was a unique sequence clone, and the pHVMK134 was a repeat sequence clone. When the pHVMK22 was used as a probe for Southern hybridization, all the powdery mildew resistance materials showed a special band of 2 kb, while all the susceptible ones not. The pHVMK22 may be applied to detect the existence of Pm21.
文摘Objective: Laser capture microdisection has become indispensable to the analysis of the difference of gene expression between human bladder transitional cell and bladder transitional cell carcinoma (BTCC). However, to obtain sufficient RNA from laser-capture microdissected cells is quite difficult. The study was designed to determinc a feasible technical routine to isolate transitional cells from bladder membrane, separate carcinoma cclls from stromal cells and to amplify the RNA isolated from laser-capture microdissected cells. Methods: Bladder transitional cell were obtained from frozen sections of bladder membrane applying LCM, by the same token, BTCC cells from frozen sections of BTCC tissue. Then RNA was extracted and linearly amplified in vitro. The expression levels of β-actin in primary total RNA and amplified RNA were detected using RT-PCR. Results: That RNA integrity was good after LCM was confirmed by control experiment Ⅰ; By control experiment Ⅱ, the correlation between the number of LCM-shooting and RNA quantity undcr arranged conditions was preliminarily confirmed. About 0.5-2.5kb RNA fragments were obtained after RNA amplification and β-actin levels were integral. Conclusion: Laser capture microdissection combined with RNA linear amplification in vitro can be successfully applied to obtain pure objective cells for research. The integrity of the amplified RNA is good and can be employed in further research.
文摘We performed this meta-analysis to evaluate the predictive value of different parameters in the sperm retrieval rate (SRR) of microdissection testicular sperm extraction (TESE) in patients with nonobstructive azoospermia (NOA). All relevant studies were searched in PubMed, Web of Science, EMBASE, Cochrane Library, and EBSCO. We chose three parameters to perform the meta-analysis: follicle-stimulating hormone (FSH), testicular volume, and testicular histopathological findings which included three patterns: hypospermatogenesis (HS), maturation arrest (MA), and Sertoli-cell-only syndrome (SCOS). If there was a threshold effect, only the area under the summary receiver operating characteristic curve (AUSROC) was calculated. Otherwise, the pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), and the diagnostic odds ratio (DOR) were also calculated. Twenty-one articles were included in our study finally. There was a threshold effect among studies investigating FSH and SCOS. The AUSROCs of FSH, testicular volume, HS, MA, and SCOS were 0.6119, 0.6389, 0.6758, 0.5535, and 0.2763, respectively. The DORs of testicular volume, HS, and MA were 1.98, 16.49, and 1.26, respectively. The sensitivities of them were 0.80, 0.30, and 0.27, while the specificities of them were 0.35, 0.98, and 0.76, respectively. The PLRs of them were 1.49, 10.63, and 1.15, respectively. And NLRs were 0.73, 0.72, and 0.95, respectively. All the investigated factors in our study had limited predictive value. However, the histopathological findings were helpful to some extent. Most patients with HS could get sperm by microdissection TESE.
文摘In 220 consecutive patients with non-obstructive azoospermia, sperm retrieval was attempted by a combination of conventional and microdissection testicular sperm extraction (TESE). For sperm retrieval, 2-3 conventional biopsies were performed followed by a microdissection TESE in cases of negative conventional biopsies. During the surgery, the vasculature of the testis was assessed using the operative microscope, and the location of positive biopsies was registered in relation to the blood supply. The overall sperm retrieval rate was 58.2%. From the initial conventional biopsies, sperm could be retrieved in 46.8% of the patients. With microdissection TESE, sperm could be retrieved from an additional 11.4% of the patients. The further use of microdissection TESE improved the sperm retrieval rate significantly (P=0.017). No significant accumulation of positive biopsies was found towards the rete testis or the main testicular vessels.
基金Supported by the Natural Science Foundation of Shanghai, No02ZB14072
文摘AIM. To study the differential gene expression profiles of target cells in primary gastric cancer and its metastatic lymph nodes using laser microdissection (LMD) in combination with cDNA microarray. METHODS: Normal gastric tissue samples from 30 healthy individuals, 36 cancer tissue samples from primary gastric carcinoma and lymph node metastasis tissue samples from 58 patients during gastric cancer resection were obtained using LMD in combination with cDNA microarray independently. After P27-based amplification, aRNA from 36 of 58 patients (group 1) with lymph node metastasis and metastatic tissue specimens from the remaining 22 patients (group 2) were applied to cDNA microarray. Semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) and imrnunohistochemical assay verified the results of microarray in group 2 and further identified genes differentially expressed in the progression of gastric cancer. RESULTS: The expression of 10 genes was up-regulated while the expression of 15 genes was down-regulated in 22 gastric carcinoma samples compared with that of genes in the normal controls. The results were confirmed at the level of mRNA and protein, and suggested that four genes (OPCML, RNASE1, YES1 and ACK1) could play a key role in the tumorigenesis and metastasis of gastric cancer. The expression pattern of 3 genes (OPCML, RNASE1 and YES1) was similar to tumor suppressor genes. For example, the expression level of these genes was the highest in normal gastric epithelium, which was decreased in primary carcinoma, and further decreased in metastatic lymph nodes. On the contrary, the expression pattern of gene ACK1 was similar to that of oncogene. Four genes were further identified as differentially expressed genes in the majority of the cases in the progression of gastric cancer. CONCLUSION: LMD in combination with cDNA microaro ray provides a unique support foe the identification of early expression profiles of differential genes and the expression pattern of 3 genes (OPCML, RNASE1 and YES1) associated with the progression of gastric cancer. Further study is needed to reveal the molecular mechanism of lymph node metastasis in patients with gastric cancer.
基金National Science Council, NSC-91-3112-B002-007, Taipei, Taiwan, China
文摘AIM: To examine the gene expression profile of gastric cancer (GC) by combination of laser capture microdissection (LCM) and microarray and to correlate the profiling with histological subtypes. METHODS: Using LCM, pure cancer cells were procured from 45 cancerous tissues. After procurement of about 5 000 cells, total RNA was extracted and the quality of RNA was determined before further amplification and hybridization. One microgram of amplified RNA was converted to cDNA and hybridized to cDNA microarray. RESULTS: Among 45 cases, only 21 were qualified for their RNAs. A total of 62 arrays were performed. These included 42 arrays for cancer (21 cases with dyeswab duplication) and 20 arrays for non-tumorous cells (10 cases with dye-swab duplication) with universal reference. Analyzed data showed 504 genes were differentially expressed and could distinguish cancerous and non-cancerous groups with more than 99% accuracy. Of the 504 genes, trefoil factors 1, 2, and 3 were in the list and their expression patterns were consistent with previous reports. Immunohistochemical staining of trefoil factor 1 was also consistent with the array data. Analyses of the tumor group with these 504 genes showed that there were 3 subgroups of GC that did not correspond to any current classification system, including Lauren's dassification. CONCLUSION: By using LCM, linear amplification of RNA, and cDNA microarray, we have identified a panel of genes that have the power to discriminate between GC and non-cancer groups. The new molecular classification and the identified novel genes in gastric carcinogenesis deserve further investigations to elucidate their clinicopathological significance.