Efficient and convenient treatment of industrial dyeing wastewater is of great significance to guarantee human and animal health.This work presented the enhanced catalytic activity at pH 3.0 of laccase immobilized on ...Efficient and convenient treatment of industrial dyeing wastewater is of great significance to guarantee human and animal health.This work presented the enhanced catalytic activity at pH 3.0 of laccase immobilized on amino-functionalized ZnFe_(2)O_(4) nanoparticles(ZnFe_(2)O_(4)-laccase)and its application for the degradation of textile dyes.Due to the existence of a large number of oxygen vacancies on the surface of the ZnFe_(2)O_(4) nanoparticles,negative ions accumulated on the magnetic carriers,which resulted in a harsh optimal pH value of the ZnFe_(2)O_(4)-laccase.Laccase activity assays revealed that the ZnFe_(2)O_(4)-laccase possessed superior pH and thermal stabilities,excellent reusability,and noticeable organic solvent tolerance.Meanwhile,the ZnFe_(2)O_(4) laccase presented efficient and sustainable degradation of high concentrations of textile dyes.The initial decoloration efficiencies of malachite green(MG),brilliant green(BG),azophloxine,crystal violet(CV),reactive blue 19(RB19),and procion red MX-5B were approximately 99.1%,95.0%,93.3%,87.4%,86.1%,and 85.3%,respectively.After 10 consecutive reuses,the degradation rates of the textile dyes still maintained about 98.2%,92.5%,83.2%,81.5%,79.8%and 65.9%,respectively.The excellent dye degradation properties indicate that the ZnFe_(2)O_(4)-laccase has a technical application in high concentrations of dyestuff treatment.展开更多
To improve the heat transfer efficiency of the coolant in lead-based fast reactors,this study optimized the configuration and rotational direction of the spacer wires in fuel assemblies to design a new-pattern fuel as...To improve the heat transfer efficiency of the coolant in lead-based fast reactors,this study optimized the configuration and rotational direction of the spacer wires in fuel assemblies to design a new-pattern fuel assembly.This study conducted detailed comparisons between traditional and new pattern fuel assembly rod bundles utilizing the open-source computational fluid dynamics platform,OpenFOAM.The results indicated that the new design may significantly reduce the pressure drop along the rod bundle,which is beneficial for lowering the pressure drop.Furthermore,this new design improved coolant mixing in the subchannels,which facilitated a more uniform temperature distribution and lower thermal gradients at the assembly outlet.These factors collectively reduced the thermal fatigue and creep in nearby internal components.Overall,the newpattern fuel assembly proposed in this study may have better heat transfer performance,thereby enhancing the Integrated Thermal-Hydraulic Factor by 48.2% compared to the traditional pattern.展开更多
BACKGROUND Complicated crown–root fracture (CRF) involves severe injury to the crown, root,and pulp, and may be accompanied by multiple root fractures. The loss of a toothhas lifelong consequences for children and te...BACKGROUND Complicated crown–root fracture (CRF) involves severe injury to the crown, root,and pulp, and may be accompanied by multiple root fractures. The loss of a toothhas lifelong consequences for children and teenagers, but the maintenance of pulphealth and the calcific healing of multiple root fractures are rarely reported in theliterature.CASE SUMMARY This case reports healing of a permanent tooth with complicated crown–root andadditional root fractures, in which pulp health was maintained. A 10-year-old girlfell and fractured the root of her maxillary left central incisor at the cervical level.After the coronal fragment was repositioned, the tooth was splinted until thetooth was no longer mobile, 2 years later. Eight years after treatment, the toothhas remained asymptomatic with vital pulp and localized gingival overgrowth.Cone-beam computed tomography revealed not only calcified healing of the CRFbut also spontaneous healing in an additional undiagnosed root fracture. Thefracture line on the enamel could not be healed by hard tissue and formed agroove in the cervical crown. It was speculated that the groove was related to thelocalized gingival overgrowth.CONCLUSION This case provides a clinical perspective of the treatment of a tooth with acomplicated CRF and an additional root fracture.展开更多
Chrysanthemum morifolium,an ornamental crop with diverse forms of inflorescence,is a good model for studying flower development in Asteraceae.However,the genetic background is complex and the mechanisms of regulating ...Chrysanthemum morifolium,an ornamental crop with diverse forms of inflorescence,is a good model for studying flower development in Asteraceae.However,the genetic background is complex and the mechanisms of regulating flower development are still unclear.Here,we identified two natural mutant lines of chrysanthemum and named them M1 and M2 according to the severity of the phenotype.Both lines showed defects in petal identity,and the petals of the M1 line had a mild phenotype:partially loss of petal identity and conversion of petals into green,leaf-like organs.The M2 line had severe phenotypes:in addition to severe petal defects,secondary inflorescences were produced in the capitulum to replace the normal ray and disc florets,which indicated a transformation of a flower meristem into an inflorescence meristem.Transcriptome sequencing of WT and M2 inflorescences was performed and found altered expression of floral organ development A,B and E class genes,where B and E class genes were significantly down-regulated.qRT-PCR analysis in both M1 and M2 lines revealed that the expression of three chrysanthemum class B genes CmAP3.1,CmAP3.2 and CmPI,was negatively correlated with phenotypic severity.This suggests that class B genes in chrysanthemum not only have conserved functions in determining petal identity but also were involved in the determinacy of the flower meristem.This study provides insights into the functions of class B genes in flower development,and is informative for dissecting the molecular mechanisms of flower development in chrysanthemum.展开更多
Brassinosteroids(BRs),a class of steroid phytohormones,play a critical role in plant growth and development.The DWF4 gene encodes a cytochrome P450 enzyme(CYP90B1),which is considered a rate-limiting enzyme in BR bios...Brassinosteroids(BRs),a class of steroid phytohormones,play a critical role in plant growth and development.The DWF4 gene encodes a cytochrome P450 enzyme(CYP90B1),which is considered a rate-limiting enzyme in BR biosynthesis.Here,we identified a homologous gene of DWF4 in chrysanthemum,CmDWF4.This gene was predicted to encode 491 amino acid residues with a molecular weight of 56.2 kDa and an isoelectric point(pI)of 9.10.Overexpression of CmDWF4 in chrysanthemum was found to significantly increase growth rate,number,and length of lateral buds.Transcriptome analysis showed that multiple xyloglucan endotransglycosylase/hydrolase(XTH)family encoding genes associated with cell wall modification were up-regulated in CmDWF4-overexpressing lines.qRT-PCR assay confirmed the up-regulation of CmXTH6,CmXTH23,and CmXTH28 in CmDWF4-overexpression line.Overall,this work establishes a mechanism by which BR biosynthetic gene CmDWF4 promotes lateral bud outgrowth in chrysanthemum,possibly through regulating cell elongation and expansion.展开更多
"Jinhua Xiangyuan"(JHXY)is a unique type of Chinese tea made from roasted oolong tea and subsequently fermented by Eurotium cristatum.In this study,we investigated the potential benefits of JHXY on obese rat..."Jinhua Xiangyuan"(JHXY)is a unique type of Chinese tea made from roasted oolong tea and subsequently fermented by Eurotium cristatum.In this study,we investigated the potential benefits of JHXY on obese rats induced by a high-sucrose high-fat diet.JHXY infusion was administered orally at 1.6 g/kg·bw(HI),0.8 g/kg·bw(MI),and 0.4 g/kg·bw(LI)per day for 10 weeks.Weight gain was significantly suppressed without affecting appetite.In the MI and HI groups,JHXY led to a significant reduction in triglycerides(TG),total cholesterol(TC),and low-density lipoprotein cholesterol(LDL-C),accompanied by an increase in high-density lipoprotein cholesterol(HDL-C)levels.A significant reduction in white adipose tissue(WAT)index and an increase in brown adipose tissue(BAT)index were observed.In MI group,the morphology of hepatocytes was restored.The abundance of Bifidobacterium was elevated,whereas the abundance of Desulfovibrio was significantly reduced.Correlation analysis revealed that the ratio of Ruminococcus to Bifidobacterium was positively correlated with HDL-C;the abundance of the Christensenellaceae_R-7_group was negatively correlated with TC and LDL-C,while positively correlated with HDL-C;the abundance of Lactobacillus was positively correlated with BAT/WAT ratio.In the MI group,the lipopolysaccharide biosynthesis pathway was predicted to be significantly weakened(p<0.05);in the HI group,the sulfur relay system pathway was attenuated,whereas the bacterial toxins pathway was augmented(p<0.05).Our research showed that JHXY has the potential to enhance metabolic health and maintain intestinal homeostasis.Optimal dosage should be a focal point in future research.展开更多
Objective:To investigate the expression of matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),and transforming growth factor beta 1(TGF beta 1)in the kidney tissue of rats with pyelonephritis and thei...Objective:To investigate the expression of matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),and transforming growth factor beta 1(TGF beta 1)in the kidney tissue of rats with pyelonephritis and their relationship with pyelonephritis by establishing a rat model of acute pyelonephritis.Methods:80 male Wistar rats were randomly divided into a control group and an experimental group,with 40 rats each.The rats of the control group were injected with and saline and those of the experimental group were injected with 10μg/mL Escherichia coli(E.coli)and saline(1:100);the solutions for both groups were administered every 3 days for 7 days.The expressions of MMP-2,MMP-9 and TGF beta 1 in the kidney tissues of rats in each group were observed.Results:The expression of MMP-9 and TGF beta 1in the kidney tissue of rat acute pyelonephritis model rats was significantly higher than those of the control group(P<0.01);the MMP-9 mRNA content in the kidney tissue of the experimental group was significantly higher than that of the control group(P<0.05);the TGF beta 1 mRNA content in the renal tissue of the experimental group increased significantly compared to the(P<0.05);MMP-2,MMP-9 and TGF beta 1 began to express in the early stage of pyelonephritis until the complete formation of renal pelvic edema.The difference between groups was statistically significant(P<0.01).Conclusion:MMP-9 and TGF beta 1 are important factors regulating renal tubular epithelial cell injury and inflammatory response.展开更多
Objective:To analyze serum vitamin D levels in patients with clear cell renal cell carcinoma(ccRCC)by flow cytometry and to investigate the relationship between hypovitaminosis D status and hyperactivation of IL-6/STA...Objective:To analyze serum vitamin D levels in patients with clear cell renal cell carcinoma(ccRCC)by flow cytometry and to investigate the relationship between hypovitaminosis D status and hyperactivation of IL-6/STAT3 signaling in ccRCC.Methods:Eighty patients diagnosed with ccRCC by our oncology department from January 2019 to December 2021 were selected as study subjects,and the control subjects were selected from patients who were receiving health check-up from our hospital(matched according to case group:control group,1:2),with 160 healthy patients.All serum samples collected from the case-control subjects were allowed to stand for 1–2 hours,centrifuged at 3000 rpm for 10 minutes,and stored in a-80°C refrigerator,from which they were removed and thawed to measure 25-hydroxyvitamin D(25(OH)D)and interleukin 6(IL-6)levels.Results:The blood calcium level of patients in the cancer group was significantly lower than that of patients in the non-cancer group,and the difference was statistically significant(P<0.05).The IL-6 level of the cancer group was significantly higher than that of the non-cancer group.In high vitamin D state,the IL-6 level of the non-cancer group was higher than that of the cancer group,and the average concentration of IL-6 in both the cancer group and the non-cancer group was significantly higher in low vitamin D state compared with high vitamin D state(P<0.05);the correlation between hypovitaminosis D status and renal Ki-67 was found to be positive.Conclusion:The results showed that serum IL-6 levels were elevated in the cancer group and circulating serum 25(OH)D levels were negatively correlated with IL-6 levels.In addition,signal transducer and activator of transcription 3(STAT3)signaling in RCC tissues was activated in ccRCC patients and in those with low vitamin D status among the cancer group and was higher than that in those with high vitamin D status.These results suggest that hypovitaminosis D status in ccRCC patients is associated with activated IL-6/STAT3 signaling and the activation of tumor proliferation markers proliferating cell nuclear antigen(PCNA),cyclin D1,and Ki-67.展开更多
基金supported by the National Natural Science Foundation of China(21471002)Scientific Research Projects of Universities in Anhui Province(2022AH040135)+1 种基金Natural Science Research Project for Anhui Universities(KJ2021A0509)Anhui Natural Science Foundation(2208085MC83).
文摘Efficient and convenient treatment of industrial dyeing wastewater is of great significance to guarantee human and animal health.This work presented the enhanced catalytic activity at pH 3.0 of laccase immobilized on amino-functionalized ZnFe_(2)O_(4) nanoparticles(ZnFe_(2)O_(4)-laccase)and its application for the degradation of textile dyes.Due to the existence of a large number of oxygen vacancies on the surface of the ZnFe_(2)O_(4) nanoparticles,negative ions accumulated on the magnetic carriers,which resulted in a harsh optimal pH value of the ZnFe_(2)O_(4)-laccase.Laccase activity assays revealed that the ZnFe_(2)O_(4)-laccase possessed superior pH and thermal stabilities,excellent reusability,and noticeable organic solvent tolerance.Meanwhile,the ZnFe_(2)O_(4) laccase presented efficient and sustainable degradation of high concentrations of textile dyes.The initial decoloration efficiencies of malachite green(MG),brilliant green(BG),azophloxine,crystal violet(CV),reactive blue 19(RB19),and procion red MX-5B were approximately 99.1%,95.0%,93.3%,87.4%,86.1%,and 85.3%,respectively.After 10 consecutive reuses,the degradation rates of the textile dyes still maintained about 98.2%,92.5%,83.2%,81.5%,79.8%and 65.9%,respectively.The excellent dye degradation properties indicate that the ZnFe_(2)O_(4)-laccase has a technical application in high concentrations of dyestuff treatment.
基金supported partly by the Ministry of Science and Technology of the People’s Republic of China(No.2020YFB1902100)the China Postdoctoral Science Foundation(No.2023M731458)+3 种基金the Science and Technology Program of Gansu ProvinceChina(No.23JRRA1099)the Postdoctoral Fellowship Program of CPSF(No.GZB20230278)financially supported by the Shanghai Municipal Commission of Economy and Informatization(No.GYQJ-2018-2-02)。
文摘To improve the heat transfer efficiency of the coolant in lead-based fast reactors,this study optimized the configuration and rotational direction of the spacer wires in fuel assemblies to design a new-pattern fuel assembly.This study conducted detailed comparisons between traditional and new pattern fuel assembly rod bundles utilizing the open-source computational fluid dynamics platform,OpenFOAM.The results indicated that the new design may significantly reduce the pressure drop along the rod bundle,which is beneficial for lowering the pressure drop.Furthermore,this new design improved coolant mixing in the subchannels,which facilitated a more uniform temperature distribution and lower thermal gradients at the assembly outlet.These factors collectively reduced the thermal fatigue and creep in nearby internal components.Overall,the newpattern fuel assembly proposed in this study may have better heat transfer performance,thereby enhancing the Integrated Thermal-Hydraulic Factor by 48.2% compared to the traditional pattern.
基金Supported by 2021 Disciplinary Construction Project in School of Dentistry,Anhui Medical University,No.2021kqxkFY05.
文摘BACKGROUND Complicated crown–root fracture (CRF) involves severe injury to the crown, root,and pulp, and may be accompanied by multiple root fractures. The loss of a toothhas lifelong consequences for children and teenagers, but the maintenance of pulphealth and the calcific healing of multiple root fractures are rarely reported in theliterature.CASE SUMMARY This case reports healing of a permanent tooth with complicated crown–root andadditional root fractures, in which pulp health was maintained. A 10-year-old girlfell and fractured the root of her maxillary left central incisor at the cervical level.After the coronal fragment was repositioned, the tooth was splinted until thetooth was no longer mobile, 2 years later. Eight years after treatment, the toothhas remained asymptomatic with vital pulp and localized gingival overgrowth.Cone-beam computed tomography revealed not only calcified healing of the CRFbut also spontaneous healing in an additional undiagnosed root fracture. Thefracture line on the enamel could not be healed by hard tissue and formed agroove in the cervical crown. It was speculated that the groove was related to thelocalized gingival overgrowth.CONCLUSION This case provides a clinical perspective of the treatment of a tooth with acomplicated CRF and an additional root fracture.
基金financially supported by the National Natural Science Foundation of China(32171855)China Agriculture Research System(CARS-23-A18),Seed Industry Project of Jiangsu Province(JBGS[2021]020)+2 种基金the China Postdoctoral Science Foundation(2019M661870)the National Key Research and Development Program of China(2020YFD1000400)a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
文摘Chrysanthemum morifolium,an ornamental crop with diverse forms of inflorescence,is a good model for studying flower development in Asteraceae.However,the genetic background is complex and the mechanisms of regulating flower development are still unclear.Here,we identified two natural mutant lines of chrysanthemum and named them M1 and M2 according to the severity of the phenotype.Both lines showed defects in petal identity,and the petals of the M1 line had a mild phenotype:partially loss of petal identity and conversion of petals into green,leaf-like organs.The M2 line had severe phenotypes:in addition to severe petal defects,secondary inflorescences were produced in the capitulum to replace the normal ray and disc florets,which indicated a transformation of a flower meristem into an inflorescence meristem.Transcriptome sequencing of WT and M2 inflorescences was performed and found altered expression of floral organ development A,B and E class genes,where B and E class genes were significantly down-regulated.qRT-PCR analysis in both M1 and M2 lines revealed that the expression of three chrysanthemum class B genes CmAP3.1,CmAP3.2 and CmPI,was negatively correlated with phenotypic severity.This suggests that class B genes in chrysanthemum not only have conserved functions in determining petal identity but also were involved in the determinacy of the flower meristem.This study provides insights into the functions of class B genes in flower development,and is informative for dissecting the molecular mechanisms of flower development in chrysanthemum.
基金This research was funded by the National Natural Science Foundation of China(31872149,32172609)China Agriculture Research System(CARS-23-A18),the“JBGS”Project of Seed Industry Revitalization in Jiangsu Province(JBGS[2021]020)the earmarked fund for Jiangsu Agricultural Industry Technology System,and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
文摘Brassinosteroids(BRs),a class of steroid phytohormones,play a critical role in plant growth and development.The DWF4 gene encodes a cytochrome P450 enzyme(CYP90B1),which is considered a rate-limiting enzyme in BR biosynthesis.Here,we identified a homologous gene of DWF4 in chrysanthemum,CmDWF4.This gene was predicted to encode 491 amino acid residues with a molecular weight of 56.2 kDa and an isoelectric point(pI)of 9.10.Overexpression of CmDWF4 in chrysanthemum was found to significantly increase growth rate,number,and length of lateral buds.Transcriptome analysis showed that multiple xyloglucan endotransglycosylase/hydrolase(XTH)family encoding genes associated with cell wall modification were up-regulated in CmDWF4-overexpressing lines.qRT-PCR assay confirmed the up-regulation of CmXTH6,CmXTH23,and CmXTH28 in CmDWF4-overexpression line.Overall,this work establishes a mechanism by which BR biosynthetic gene CmDWF4 promotes lateral bud outgrowth in chrysanthemum,possibly through regulating cell elongation and expansion.
基金funded by Applied Basic Research Project of COFCO(2017-C1-T06)Yunnan Province Science and Technology Project(202102AE090038).
文摘"Jinhua Xiangyuan"(JHXY)is a unique type of Chinese tea made from roasted oolong tea and subsequently fermented by Eurotium cristatum.In this study,we investigated the potential benefits of JHXY on obese rats induced by a high-sucrose high-fat diet.JHXY infusion was administered orally at 1.6 g/kg·bw(HI),0.8 g/kg·bw(MI),and 0.4 g/kg·bw(LI)per day for 10 weeks.Weight gain was significantly suppressed without affecting appetite.In the MI and HI groups,JHXY led to a significant reduction in triglycerides(TG),total cholesterol(TC),and low-density lipoprotein cholesterol(LDL-C),accompanied by an increase in high-density lipoprotein cholesterol(HDL-C)levels.A significant reduction in white adipose tissue(WAT)index and an increase in brown adipose tissue(BAT)index were observed.In MI group,the morphology of hepatocytes was restored.The abundance of Bifidobacterium was elevated,whereas the abundance of Desulfovibrio was significantly reduced.Correlation analysis revealed that the ratio of Ruminococcus to Bifidobacterium was positively correlated with HDL-C;the abundance of the Christensenellaceae_R-7_group was negatively correlated with TC and LDL-C,while positively correlated with HDL-C;the abundance of Lactobacillus was positively correlated with BAT/WAT ratio.In the MI group,the lipopolysaccharide biosynthesis pathway was predicted to be significantly weakened(p<0.05);in the HI group,the sulfur relay system pathway was attenuated,whereas the bacterial toxins pathway was augmented(p<0.05).Our research showed that JHXY has the potential to enhance metabolic health and maintain intestinal homeostasis.Optimal dosage should be a focal point in future research.
基金primarily financially supported by the National Natural Science Foundation of China(No.51871245)the National Key R&D Program of China(No.2017YFE0301505)State Key Laboratory for Powder Metallurgy as well as State Key Laboratory of Light Weight and High Strength Structural Materials,Central South University,China。
基金Health Commission of Hebei Province:Chuanxiong Extract Improves Inflammatory Response in Rats with Pyelonephritis through IL-6/STAT3 Signaling Pathway(Project number:20231486)。
文摘Objective:To investigate the expression of matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),and transforming growth factor beta 1(TGF beta 1)in the kidney tissue of rats with pyelonephritis and their relationship with pyelonephritis by establishing a rat model of acute pyelonephritis.Methods:80 male Wistar rats were randomly divided into a control group and an experimental group,with 40 rats each.The rats of the control group were injected with and saline and those of the experimental group were injected with 10μg/mL Escherichia coli(E.coli)and saline(1:100);the solutions for both groups were administered every 3 days for 7 days.The expressions of MMP-2,MMP-9 and TGF beta 1 in the kidney tissues of rats in each group were observed.Results:The expression of MMP-9 and TGF beta 1in the kidney tissue of rat acute pyelonephritis model rats was significantly higher than those of the control group(P<0.01);the MMP-9 mRNA content in the kidney tissue of the experimental group was significantly higher than that of the control group(P<0.05);the TGF beta 1 mRNA content in the renal tissue of the experimental group increased significantly compared to the(P<0.05);MMP-2,MMP-9 and TGF beta 1 began to express in the early stage of pyelonephritis until the complete formation of renal pelvic edema.The difference between groups was statistically significant(P<0.01).Conclusion:MMP-9 and TGF beta 1 are important factors regulating renal tubular epithelial cell injury and inflammatory response.
基金funded by the Health Commission of Hebei Province under the project Chuanxiong Extract Improves Inflammatory Response in Rats with Pyelonephritis Through IL-6/STAT3 Signaling Pathway(Project Number:20231486).
文摘Objective:To analyze serum vitamin D levels in patients with clear cell renal cell carcinoma(ccRCC)by flow cytometry and to investigate the relationship between hypovitaminosis D status and hyperactivation of IL-6/STAT3 signaling in ccRCC.Methods:Eighty patients diagnosed with ccRCC by our oncology department from January 2019 to December 2021 were selected as study subjects,and the control subjects were selected from patients who were receiving health check-up from our hospital(matched according to case group:control group,1:2),with 160 healthy patients.All serum samples collected from the case-control subjects were allowed to stand for 1–2 hours,centrifuged at 3000 rpm for 10 minutes,and stored in a-80°C refrigerator,from which they were removed and thawed to measure 25-hydroxyvitamin D(25(OH)D)and interleukin 6(IL-6)levels.Results:The blood calcium level of patients in the cancer group was significantly lower than that of patients in the non-cancer group,and the difference was statistically significant(P<0.05).The IL-6 level of the cancer group was significantly higher than that of the non-cancer group.In high vitamin D state,the IL-6 level of the non-cancer group was higher than that of the cancer group,and the average concentration of IL-6 in both the cancer group and the non-cancer group was significantly higher in low vitamin D state compared with high vitamin D state(P<0.05);the correlation between hypovitaminosis D status and renal Ki-67 was found to be positive.Conclusion:The results showed that serum IL-6 levels were elevated in the cancer group and circulating serum 25(OH)D levels were negatively correlated with IL-6 levels.In addition,signal transducer and activator of transcription 3(STAT3)signaling in RCC tissues was activated in ccRCC patients and in those with low vitamin D status among the cancer group and was higher than that in those with high vitamin D status.These results suggest that hypovitaminosis D status in ccRCC patients is associated with activated IL-6/STAT3 signaling and the activation of tumor proliferation markers proliferating cell nuclear antigen(PCNA),cyclin D1,and Ki-67.
