Present and future of new systemic therapies for early and intermediate stages of hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is a high mortality neoplasm which usually appears on a cirrhotic liver.The therapeutic arsenal and subsequent prognostic outlook are intrinsically linked to the HCC stage at diagnosis.Notwithstanding the current deployment of treatments with curative intent(liver resection/local ablation and liver transplantation)in early and intermediate stages,a high rate of HCC recurrence persists,underscoring a pivotal clinical challenge.Emergent systemic therapies(ST),particularly immunotherapy,have demonstrate promising outcomes in terms of increase overall survival,but they are currently bound to the advanced stage of HCC.This review provides a comprehensive analysis of the literature,encompassing studies up to March 10,2024,evaluating the impact of novel ST in the early and intermediate HCC stages,specially focusing on the findings of neoadjuvant and adjuvant regimens,aimed at increasing significantly overall survival and recurrence-free survival after a treatment with curative intent.We also investigate the potential role of ST in enhancing the downstaging rate for the intermediate-stage HCC initially deemed ineligible for treatment with curative intent.Finally,we critically discuss about the current relevance of the results of these studies and the encouraging future implications of ST in the treatment schedules of early and intermediate HCC stages.

Manure substitution improves maize yield by promoting soil fertility and mediating the microbial community in lime concretion black soil

Synthetic nitrogen(N)fertilizer has made a great contribution to the improvement of soil fertility and productivity,but excessive application of synthetic N fertilizer may cause agroecosystem risks,such as soil acidification,groundwater contamination and biodiversity reduction.Meanwhile,organic substitution has received increasing attention for its ecologically and environmentally friendly and productivity benefits.However,the linkages between manure substitution,crop yield and the underlying microbial mechanisms remain uncertain.To bridge this gap,a three-year field experiment was conducted with five fertilization regimes:i)Control,non-fertilization;CF,conventional synthetic fertilizer application;CF_(1/2)M_(1/2),1/2 N input via synthetic fertilizer and 1/2 N input via manure;CF_(1/4)M_(3/4),1/4 N input synthetic fertilizer and 3/4 N input via manure;M,manure application.All fertilization treatments were designed to have equal N input.Our results showed that all manure substituted treatments achieved high soil fertility indexes(SFI)and productivities by increasing the soil organic carbon(SOC),total N(TN)and available phosphorus(AP)concentrations,and by altering the bacterial community diversity and composition compared with CF.SOC,AP,and the soil C:N ratio were mainly responsible for microbial community variations.The co-occurrence network revealed that SOC and AP had strong positive associations with Rhodospirillales and Burkholderiales,while TN and C:N ratio had positive and negative associations with Micromonosporaceae,respectively.These specific taxa are implicated in soil macroelement turnover.Random Forest analysis predicted that both biotic(bacterial composition and Micromonosporaceae)and abiotic(AP,SOC,SFI,and TN)factors had significant effects on crop yield.The present work strengthens our understanding of the effects of manure substitution on crop yield and provides theoretical support for optimizing fertilization strategies.

Diagnosis of poorly differentiated adenocarcinoma of the stomach by confocal laser endomicroscopy:A case report

BACKGROUND In recent years,confocal laser endomicroscopy(CLE)has become a new endoscopic imaging technology at the microscopic level,which is extensively performed for real-time in vivo histological examination.CLE can be performed to distinguish benign from malignant lesions.In this study,we diagnosed using CLE an asymptomatic patient with poorly differentiated gastric adenocarcinoma.CASE SUMMARY A 63-year-old woman was diagnosed with gastric mucosal lesions,which may be gastric cancer,in the small curvature of the stomach by gastroscopy.She consented to undergo CLE for morphological observation of the gastric mucosa.Through the combination of CLE diagnosis and postoperative pathology,the intraoperative CLE diagnosis was considered to be reliable.According to our experience,CLE can be performed as the first choice for the diagnosis of gastric cancer.CONCLUSION CLE has several advantages over pathological diagnosis.We believe that CLE has great potential in the diagnosis of benign and malignant gastric lesions.

Impact pressure of waves generated by landslides on bank slopes

Impulse waves that are generated by landslides in narrow reservoir areas threaten the stability of buildings and bank slopes.To discuss the action process and evolution law of the wave pressure on bank slopes,a three-dimensional physical model test that considers impulse waves generated by landslides was performed,and factors including landslide width,thickness,slope angles of the sliding surface,and bank slope angle were considered.Based on wave forms on the bank slopes,wave pressure curve characteristics,and peak value,the action process of wave pressure could be divided into the following stages:maximum pulsating pressure stage,wave impact stage(when waves break),and stationary pulsation stage.It was found that wave breaking is dependent on the value of the surf similarity parameterξ.The distribution pattern of impact pressure decays linearly on both sides of the maximum impact pressure point,and the attenuation degree decreases when it attains 40%of the maximum value.Thus,it is proposed that the prediction formula for the maximum effective impact pressure of the bank slope be related to the reciprocal of wave steepness,relative water depth,and slope rate.The prediction formula provides strong theoretical support for early safety warning and for predicting the bank slope under impulse waves generated by landslides.

Single-cell profiling of the pig cecum at various developmental stages

The gastrointestinal tract is essential for food digestion,nutrient absorption,waste elimination,and microbial defense.Single-cell transcriptome profiling of the intestinal tract has greatly enriched our understanding of cellular diversity,functional heterogeneity,and their importance in intestinal tract development and disease.Although such profiling has been extensively conducted in humans and mice,the single-cell gene expression landscape of the pig cecum remains unexplored.Here,single-cell RNA sequencing was performed on 45572 cells obtained from seven cecal samples in pigs at four different developmental stages(days(D)30,42,150,and 730).Analysis revealed 12 major cell types and 38 subtypes,as well as their distinctive genes,transcription factors,and regulons,many of which were conserved in humans.An increase in the relative proportions of CD8^(+)T and Granzyme A(low expression)natural killer T cells(GZMA^(low)NKT)cells and a decrease in the relative proportions of epithelial stem cells,Tregs,RHEX^(+)T cells,and plasmacytoid dendritic cells(pDCs)were noted across the developmental stages.Moreover,the post-weaning period exhibited an up-regulation in mitochondrial genes,COX2 and ND2,as well as genes involved in immune activation in multiple cell types.Cell-cell crosstalk analysis indicated that IBP6^(+)fibroblasts were the main signal senders at D30,whereas IBP6^(−)fibroblasts assumed this role at the other stages.NKT cells established interactions with epithelial cells and IBP6^(+)fibroblasts in the D730 cecum through mediation of GZMA-F2RL1/F2RL2 pairs.This study provides valuable insights into cellular heterogeneity and function in the pig cecum at different development stages.

Novel method for identifying the stages of discharge underwater based on impedance change characteristic

It is difficult to determine the discharge stages in a fixed time of repetitive discharge underwater due to the arc formation process being susceptible to external environmental influences. This paper proposes a novel underwater discharge stage identification method based on the Strong Tracking Filter(STF) and impedance change characteristics. The time-varying equivalent circuit model of the discharge underwater is established based on the plasma theory analysis of the impedance change characteristics and mechanism of the discharge process. The STF is used to reduce the randomness of the impedance of repeated discharges underwater, and then the universal identification resistance data is obtained. Based on the resistance variation characteristics of the discriminating resistance of the pre-breakdown, main, and oscillatory discharge stages, the threshold values for determining the discharge stage are obtained. These include the threshold values for the resistance variation rate(K) and the moment(t).Experimental and error analysis results demonstrate the efficacy of this innovative method in discharge stage determination, with a maximum mean square deviation of Scrless than 1.761.

Nutritional Composition and Distribution of Heracleum dissectum Ledeb.at Different Growth Stages

Heracleum dissectum Ledeb.(HD)with high edible and medicinal values is distributed in Northeast Asia.To provide a reference for selecting appropriate parts and periods to harvest HD,the nutrient contents in different parts of HD at three growth stages were determined.The nutrients being determined are essential nutrients closely related to human needs,and the crude protein content of HD was 3.58‒28.43 g•100 g^(-1)DW(dry weight),sugar content was 2.33‒30.92 g•100 g^(-1)DW,and soluble dietary fiber content was 9.71‒26.09 g•100 g^(-1)DW.Crude protein was mainly distributed in leaf blades and inflorescences,sugar was primarily found in roots,and dietary fiber was predominantly located in leaves.From the leaf development stage to the flowering stage and then to the late seed harvest stage,the content of moisture,crude protein and soluble sugar first decreased and then increased.In contrast,the content of dietary fiber and crude fat first increased and then decreased.

Autophagy-dependent ferroptosis may play a critical role in early stages of diabetic retinopathy

Diabetic retinopathy(DR),as one of the most common and significant microvascular complications of diabetes mellitus(DM),continues to elude effective targeted treatment for vision loss despite ongoing enrichment of the understanding of its pathogenic mechanisms from perspectives such as inflammation and oxidative stress.Recent studies have indicated that characteristic neuroglial degeneration induced by DM occurs before the onset of apparent microvascular lesions.In order to comprehensively grasp the early-stage pathological changes of DR,the retinal neurovascular unit(NVU)will become a crucial focal point for future research into the occurrence and progression of DR.Based on existing evidence,ferroptosis,a form of cell death regulated by processes like ferritinophagy and chaperone-mediated autophagy,mediates apoptosis in retinal NVU components,including pericytes and ganglion cells.Autophagy-dependent ferroptosis-related factors,including BECN1 and FABP4,may serve as both biomarkers for DR occurrence and development and potentially crucial targets for future effective DR treatments.The aforementioned findings present novel perspectives for comprehending the mechanisms underlying the early-stage pathological alterations in DR and open up innovative avenues for investigating supplementary therapeutic strategies.

Lymph node dissection does not affect the survival of patients with tumor node metastasis stages Ⅰ and Ⅱ colorectal cancer

BACKGROUND The effect of the number of lymph node dissections(LNDs)during radical resection for colorectal cancer(CRC)on overall survival(OS)remains controver-sial.AIM To investigate the association between the number of LNDs and OS in patients with tumor node metastasis(TNM)stage Ⅰ–Ⅱ CRC undergoing radical resection.METHODS Patients who underwent radical resection for CRC at a single-center hospital between January 2011 and December 2021 were retrospectively analyzed.Cox regression analyses were performed to identify the independent predictors of OS at different T stages.RESULTS A total of 2850 patients who underwent laparoscopic radical resection for CRC were enrolled.At stage T1,age[P<0.01,hazard ratio(HR)=1.075,95%confidence interval(CI):1.019-1.134]and tumour size(P=0.021,HR=3.635,95%CI:1.210-10.917)were independent risk factors for OS.At stage T2,age(P<0.01,HR=1.064,95%CI:1.032-1.098)and overall complications(P=0.012,HR=2.297,95%CI:1.200-4.397)were independent risk factors for OS.At stage T3,only age(P<0.01,HR=1.047,95%CI:1.027-1.066)was an independent risk factor for OS.At stage T4,age(P<0.01,HR=1.057,95%CI:1.039-1.075)and body mass index(P=0.034,HR=0.941,95%CI:0.890-0.995)were independent risk factors for OS.However,there was no association between LNDs and OS in stages Ⅰ and Ⅱ.CONCLUSION The number of LDNs did not affect the survival of patients with TNM stages Ⅰ and Ⅱ CRC.Therefore,insufficient LNDs should not be a cause for alarm during the surgery.

Estimating the Yield Loss of Winter Wheat from Drought in the United States Southern Plains Region as Influenced by El Niño-Southern Oscillation (ENSO)

Wheat (Triticum aestivum L.) production is a major economic activity in most regional and rural areas in the Southern Plains, a semi-arid region of the United States. This region is vulnerable to drought and is projected to experience a drier climate in the future. Since the interannual variability in climate in this region is linked to an ocean-atmospheric phenomenon, called El Niño-Southern Oscillation (ENSO), droughts in this region may be associated with ENSO. Droughts that occur during the critical growth phases of wheat can be extremely costly. However, the losses due to an impending drought can be minimized through mitigation measures if it is predicted in advance. Predicting the yield loss from an imminent drought is crucial for stakeholders. One of the reliable ways for such prediction is using a plant physiology-based agricultural drought index, such as Agricultural Reference Index for Drought (ARID). This study developed ENSO phase-specific, ARID-based models for predicting the drought-induced yield loss for winter wheat in this region by accounting for its phenological phase-specific sensitivity to drought. The reasonable values of the drought sensitivity coefficients of the yield model for each ENSO phase (El Niño, La Niña, or Neutral) indicated that the yield models reflected reasonably well the phenomena of water stress decreasing the winter wheat yields in this region during different ENSO phases. The values of various goodness-of-fit measures used, including the Nash-Sutcliffe Index (0.54 to 0.67), the Willmott Index (0.82 to 0.89), and the percentage error (20 to 26), indicated that the yield models performed fairly well at predicting the ENSO phase-specific loss of wheat yields from drought. This yield model may be useful for predicting yield loss from drought and scheduling irrigation allocation based on the phenological phase-specific sensitivity to drought as impacted by ENSO.

Green manuring facilitates bacterial community dispersal across different compartments of subsequent tobacco

Green manure–crop rotation is a sustainable approach to protect crops against diseases and improve yield.However,the mechanism by which green manuring manipulates the crop-associated microbial community remains to be elucidated.In this study,we explored the horizontal processes of bacterial communities in different compartments of the soil–root interface(bulk soil,rhizosphere soil,rhizoplane and endosphere)of tobacco by performing a field experiment including four rotation practices,namely,tobacco rotated with smooth vetch,ryegrass,radish,and winter fallow(without green manure).Results showed that the co-occurrence networks constructed by adjacent compartments of the soil–root interface with green manuring had more edges than without green manuring,indicating dramatic microbial interactions.Green manuring increased the dispersal-niche continuum index between bulk soil and other compartments,indicating that it facilitated the horizontal dispersal of microbes.For the different green manuring practices,the neutral community model explained 24.6–27.6%of detection frequency for bacteria,and at least one compartment under each practice had a normalized stochasticity ratio higher than the 50%boundary point,suggesting that the deterministic and stochastic processes jointly shaped the tobacco microbiome.In conclusion,green manuring generally facilitates bacterial community dispersal across different compartments and enhances potential interactions among adjacent compartments.This study provides empirical evidence for understanding the microbiome assembly under green manure–crop rotation.

Carbon sequestration rate,nitrogen use efficiency and rice yield responses to long-term substitution of chemical fertilizer by organic manure in a rice–rice cropping system

Combined application of chemical fertilizers with organic amendments was recommended as a strategy for improving yield,soil carbon storage,and nutrient use efficiency.However,how the long-term substitution of chemical fertilizer with organic manure affects rice yield,carbon sequestration rate(CSR),and nitrogen use efficiency(NUE)while ensuring environmental safety remains unclear.This study assessed the long-term effect of substituting chemical fertilizer with organic manure on rice yield,CSR,and NUE.It also determined the optimum substitution ratio in the acidic soil of southern China.The treatments were:(i)NPK0,unfertilized control;(ii)NPK1,100%chemical nitrogen,phosphorus,and potassium fertilizer;(iii)NPKM1,70%chemical NPK fertilizer and 30%organic manure;(iv)NPKM2,50%chemical NPK fertilizer and 50%organic manure;and(v)NPKM3,30%chemical NPK fertilizer and 70%organic manure.Milk vetch and pig manure were sources of manure for early and late rice seasons,respectively.The result showed that SOC content was higher in NPKM1,NPKM2,and NPKM3 treatments than in NPK0 and NPK1 treatments.The carbon sequestration rate increased by 140,160,and 280%under NPKM1,NPKM2,and NPKM3 treatments,respectively,compared to NPK1 treatment.Grain yield was 86.1,93.1,93.6,and 96.5%higher under NPK1,NPKM1,NPKM2,and NPKM3 treatments,respectively,compared to NPK0 treatment.The NUE in NPKM1,NPKM2,and NPKM3 treatments was higher as compared to NPK1 treatment for both rice seasons.Redundancy analysis revealed close positive relationships of CSR with C input,total N,soil C:N ratio,catalase,and humic acids,whereas NUE was closely related to grain yield,grain N content,and phenol oxidase.Furthermore,CSR and NUE negatively correlated with humin acid and soil C:P and N:P ratios.The technique for order of preference by similarity to ideal solution(TOPSIS)showed that NPKM3 treatment was the optimum strategy for improving CSR and NUE.Therefore,substituting 70%of chemical fertilizer with organic manure could be the best management option for increasing CSR and NUE in the paddy fields of southern China.

Translocation and Distribution of Carbon-Nitrogen in Relation to Rice Yield and Grain Quality as Affected by High Temperature at Early Panicle Initiation Stage

Due to climate change, extreme heat stress events have become more frequent, adversely affecting rice yield and grain quality. The accumulation and translocation of dry matter and nitrogen substances are essential for rice yield and grain quality. To assess the impact of high temperature stress(HTS) at the early panicle initiation(EPI) stage on the accumulation, transportation, and distribution of dry matter and nitrogen substances in various organs of rice, as well as the resulting effects on rice yield and grain quality, pot experiments were conducted using an indica rice cultivar Yangdao 6(YD6) and a japonica rice cultivar Jinxiangyu 1(JXY1) under both normal temperature(32 ℃/26 ℃) and high temperature(38 ℃/29 ℃) conditions. The results indicated that exposure to HTS at the EPI stage significantly decreased rice yield by reducing spikelet number per panicle, grain-filling rate, and grain weight. However, it improved the nutritional quality of rice grains by increasing protein and amylose contents. The reduction in nitrogen and dry matter accumulation accounted for the changes in spikelet number per panicle, grain-filling rate, and grain size. Under HTS, the decrease in nitrogen accumulation accompanied by the reduction in dry matter may be due to the down-regulation of leaf net photosynthesis and senescence, as evidenced by the decrease in nitrogen content. Furthermore, the decrease in sink size limited the translocation of dry matter and nitrogen substances to grains, which was closely related to the reduction in grain weight and the deterioration of grain quality. These findings significantly contribute to our understanding of the mechanisms of HTS on grain yield and quality formation from the perspective of dry matter and nitrogen accumulation and translocation. Further efforts are needed to improve the adaptability of rice varieties to climate change in the near future.

Prevention and Control Effects of New Biological Formulations Sprayed by Different Methods on Proceras venosatus (Walker) and Tryporyza intacta Snellen in Middle and Late Stages

To screen new biological formulations, accurate and efficient application technology, field test was conducted with 72% Bacillus thuringiensis(Bt)·monosultap WP, 8% lambda-cyhalothrin·emamectin benzoate SA and 3.6% lambda-cyhalothrin·Bt SA through manual spraying and unmanned aerial vehicle spraying. Test results and comprehensive evaluation analysis demonstrated that 72% Bt·monosultap WP and 8%lambda-cyhalothrin·emamectin benzoate SA by manual spraying and unmanned aerial vehicle spraying had good prevention and control effect on strains and internodes damaged by Tryporyza intacta Snellen in middle and late stage, which were ideal new biological formulations with high effi-ciency and low risk for prevention and control of T. intacta in middle and late stage, and could be popularized in sugarcane area. 72% Bt·monosultap WP at the dose of 3 000 g/hectare and 8% lambda-cyhalothrin·emamectin benzoate SA at the dose of 750 mL/hectare could be sprayed in mid September at the peak occurrence period of the forth and fifth generations of T. intacta and Proceras venosatus. Agents were diluted with 900 kg water per hectare, and manually sprayed with electric knapsack sprayer; or agents were diluted with 15 kg of Haoyang aerial control special addi-tives and water per hectare, and sprayed with unmanned aerial vehicle. The control effect against borer-damaged strain rate was above 81.3% and that against borer-damaged internode rate was above 88.6%.

Late Paleoproterozoic-Neoproterozoic Multi-rifting Events Accompanied by Four Stages of Magmatism in the North China Craton and Their Geological Significance

An important Paleoproterozoic mobile belt event took place in the North China Craton(NCC),termed the Hutuo Movement.This has been interpreted as a cratonic reworking event with rifting-subduction-collision

Production of Biogas from Olive Mill Waste Waters Treated by Cow Manure

The olive mill waste waters (OMWW) generated from olive oil extraction is a major environmental concern since they are characterized by their role as a pollutant (high organic and mineral matters) and their pH acid. The aim of this study was to valorize (OMWW) by anaerobic fermentation in the presence of cow manure in order to produce biogas and reduce their toxic load. Many tests were carried out by fermenting (OMWW) with polyphenols in the presence of cow manure in thermophile temperatures. The performance of this treatment was valuated through measurements of biogas production and by the determination of different parameters of fermented media (pH, volume of the biogas and polyphenols).

Understanding changes in volatile compounds and fatty acids of Jincheng orange peel oil at different growth stages using GC-MS

Jincheng orange(Citrus sinensis Osbeck)is widely grown in Chongqing,China,and is commonly consumed because of its characteristic aroma contributed by the presence of diverse volatile compounds.The changes in aroma during the development and maturation of fruit are indicators for ripening and harvest time.However,the influence of growth stages on the volatile compounds in Jincheng orange remains unclear.In addition,volatiles originate from fatty acids,most of which are the precursors of volatile substances.On this basis,gas chromatography-mass spectrometry(GC-MS)was performed to elaborate the changes in volatile constituents and fatty acids as precursors.This study tested proximately 60 volatiles and 8 fatty acids at 9 growth and development stages(AF1-AF9).Of those compounds,more than 92.00%of total volatiles and 87.50%of fatty acids were terpenoid and saturated fatty acids,respectively.As shown in the PCA plot,the AF5,AF6,and AF9 stages were confirmed as completely segregated and appeared different.In addition,most of the volatiles and fatty acids first increased at the beginning of the development stage,then decreased from the AF6 development stage,and finally increased at the AF9 maturity stage.Moreover,the highest contents of terpenoid,alcohols,aldehydes,ketones,and saturated fatty acids in Jincheng orange peel oil were d-limonene,linalool,octanal,cyclohexanone,and stearic acid during development stages,respectively.Our results found that the growth stages significantly affected the volatile constituents and precursors in Jincheng orange peel oil.

Mid-term results of sub-trochanteric valgus osteotomy for symptomatic late stages Legg-Calvé-Perthes disease

BACKGROUND The treatment of late stages of Legg-Calvé-Perthes disease(LCPD)is controversial.Although the concept of femoral head containment is a well-established technique of treatment,its use remains debatable in the late stages of the disease,as it does not improve symptoms in terms of limb length discrepancy and gait.AIM To assess the results of subtrochanteric valgus osteotomy in symptomatic patients with late-stage Perthes disease.METHODS From 2000 to 2007,36 symptomatic patients with late stage of Perthes disease were surgically treated with subtrochanteric valgus osteotomy and followed-up for 8 to 11 years using the IOWA score and range of motion(ROM)variables.The Mose classification was also assessed at the last follow-up to reflect possible remodeling.The patients were 8 years old or older at the time of surgery,in the post-fragmentation stage,and complaining of pain,limited ROM,Trendelenburg gait,and/or abductor weakness.RESULTS The preoperative IOWA score(average:53.3)markedly improved at the 1-year post follow-up period(average:85.41)and then slightly improved at the last follow-up(average:89.4)(P value<0.05).ROM improved,with internal rotation increased on average by 22°(from 10°preoperatively to 32°postoperatively)and abduction increased on average by 15.9°(from 25°preoperatively to 41°postoperatively).The mean Mose deviation of femoral heads was 4.1 mm at the end of the follow-up period.The tests used were the paired t-test and Pearson correlation test,where the level of significance was a P value less than 0.05.CONCLUSION Subtrochanteric valgus osteotomy can be a good option for symptomatic relief in patients with late-stage of LCPD.

Rectal Cancer Stages T2 and T3 Identification Based on Asymptotic Hybrid Feature Maps

Many existing intelligent recognition technologies require huge datasets for model learning.However,it is not easy to collect rectal cancer images,so the performance is usually low with limited training samples.In addition,traditional rectal cancer staging is time-consuming,error-prone,and susceptible to physicians’subjective awareness as well as professional expertise.To settle these deficiencies,we propose a novel deep-learning model to classify the rectal cancer stages of T2 and T3.First,a novel deep learning model(RectalNet)is constructed based on residual learning,which combines the squeeze-excitation with the asymptotic output layer and new cross-convolution layer links in the residual block group.Furthermore,a two-stage data augmentation is designed to increase the number of images and reduce deep learning’s dependence on the volume of data.The experiment results demonstrate that the proposed method is superior to many existing ones,with an overall accuracy of 0.8583.Oppositely,other traditional techniques,such as VGG16,DenseNet121,EL,and DERNet,have an average accuracy of 0.6981,0.7032,0.7500,and 0.7685,respectively.

RT-qPCR normalization of reference genes in different lifehistory stages of Gracilaria vermiculophylla(Rhodophyta)

The quantitative realtime polymerase chain reaction(RT-qPCR)is a powerful and sensitive method to measure expression of targeted gene but it highly relies on the use of suitable reference genes for data normalization.We evaluated the expressions of 8 housekeeping genes:18S ribosomal rDNA(18S rDNA),28S ribosomal r DNA(28S rDNA),rubisco large subunit(rbc-L),β-actin(ACT),glyceraldehyde-3-phosphate dehydrogenase(GAPDH),elongation factor 1(EF1),β-tubulin(Tub B),and P-phycoerythrin B(PEB),to select the suitable reference genes for different life-history stages(tetrasporophyte,carposporophyte,and male/female gametophyte)of Gracilaria vermiculophylla by absolute quantitative method.Softwares geNorm and BestKeeper were used to verify the results acquired from copy number analysis.Results show that the expression of identified reference genes varied in comparing groups composed of different type of life stages.It is suggested that 18S rDNA and TubB could be used for highly complex samples composed of mixed ploidy and phases.18S rDNA and 28S rDNA were also preferred for using among the matured isomorphic samples.But for samples with different maturities,TubB and ACT were recommended for tetrasporophytes and gametophytes respectively.