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A retrospective study of PD-L1 Immunohistochemistry for hepatocellular carcinoma

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iLIVER 2022年第3期1卷 187-193页

作者： Min Du Yumeng Cai Yanrui Pang Yuan Ji Pathology Department Huadong HospitalFudan UniversityShanghai200040China Pathology Department Zhongshan HospitalFudan UniversityShanghai200032China

Background:Programmed cell death ligand 1(PD-L1)expression plays an important role in selecting patients with hepatocellular carcinoma who will benefit from anti-programmed cell death 1(PD-1)/PD-L1 monotherapy and directing those with lower levels to alternative treatments.Methods:In total,156 hepatocellular carcinoma tumors were stained with three PD-L1 Immunohistochemistry assays(SP142,28-8,and E1L3N).Two junior pathologists and one senior pathologist evaluated the pathological slides and recorded the percentages of tumor and immune cells stained at any intensity.Results:Analytical comparisons demonstrated that the anti-PD-L1 assay SP142 is a significantly stronger reagent that stains a higher percent of both tumor and immune cells,while 28-8 and E1L3N behave similarly to each other.The correlation coefficients of the three assays ranged from 0.77 to 0.92 for tumor cells and from 0.66 to 0.75 for immune cells.Next,1%and 5%tumor cell staining cutoffs were designated using various combinations of assays and cutoffs.Across all threereagents,14 samples showedconcordanceabove the5%threshold,eight showed concordancewithinthe range of 1%to 5%interval,and 93 showed concordance below the 1%threshold.The remaining 41 samples showed a combination of discordant results across all three reagents.High PD-L1 expression in tumor and immune cells tended to not be recurrent.Conclusions:The anti-PD-L1 assays 28-8 and E1L3N were closely aligned with regard to staining tumor and immune cells,whereas SP142 showed higher percentages of staining for both cell types.All three assays indicated greater variability in immune cell staining than in tumor cells.

关键词： Programmed cell death ligand 1 Immunohistochemistry Hepatocellular carcinoma Immune checkpoint inhibitor

Study of penetration mechanism of labrasol on rabbit cornea by Ussing chamber, RT-PCR assay, Western blot and Immunohistochemistry

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Asian Journal of Pharmaceutical Sciences 2019年第3期14卷 329-339页

作者： Pan Guo Nan Li Lili Fan Jun Lu Boying Liu Bing Zhang Yumei Wu Zhidong Liu Jiawei Li Jiaxin Pi Dongli Qi Tianjin State Key Laboratory of Modern Chinese Medicine The Institute of Traditional Chinese MedicineTianjinUniversity of Traditional Chinese MedicineNo.88Yuquan RoadTianjin300193China Engineering Research Center of Modern Chinese Medicine Discovery and Preparation Tianjin University ofTraditional Chinese MedicineNo.88Yuquan RoadTianjin300193China School of Pharmaceutical Science and Technology Tianjin UniversityTianjin300193China

Labrasol, as a non-ionic surfactant, can enhance the permeation and absorption of drugs, and is extensively used in topical, transdermal, and oral pharmaceutical preparations as an emulsifier and absorption enhancer. Recent studies in our laboratory have indicated that labrasol has a strong absorption enhancing effect on different types of drugs in vitro and in vivo. This study was performed to further elucidate the action mechanism of labrasol on the corneal penetration. In this research, the fluorescein sodium, a marker of passive paracellular transport of tight junction, was selected as the model drug to assess the effect of labrasol on in vitro corneal permeability. To investigate the continuous and real-time influence of labrasol on the membrane permeability and integrity, the Ussing chamber system was applied to monitor the electrophysiological parameters. And, furthermore, we elucidated the effect of labrasol on excised cornea at the molecular level by application of RT-PCR, Western blot, and immunohistochemical staining. The results indicated that labrasol obviously enhance the transcorneal permeability of fluorescein sodium, and the enhancement was realized by interacting with and down-regulating the associated proteins, such as Factin, claudin-1 and β-catenin, which were contributed to cell-cell connections, respectively.

关键词： Enhanced corneal permeation Tight junctions associated proteins Labrasol Ussing chamber Western blot Immunohistochemistry

Molecular genetics and Immunohistochemistry characterization of uncommon and recently described renal cell carcinomas

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Chinese Journal of Cancer Research 2016年第1期28卷 29-49页

作者： Qiu Rao Qiu-Yuan Xia Liang Cheng Xiao-Jun Zhou Department of Pathology Jinling Hospital Nanjing University School of Medicine Nanjing China Department of Pathology and Laboratory Indiana University School of Medicine Indianapolis IN USA

Renal cell carcinoma （RCC） compromises multiple types and has been emerging dramatically over the recent several decades. Advances and consensus have been achieved targeting common RCCs, such as clear cell carcinoma, papillary RCC and chromophobe RCC. Nevertheless, little is known on the characteristics of several newly-identified RCCs, including clear cell （tubulo） papillary RCC, Xpl 1 translocation RCC, t（6;11） RCC, succinate dehydrogenase （SDH）-deficient RCC, acquired cystic disease- associated RCC, hereditary leiomyomatosis RCC syndrome-associated RCC, ALK translocation RCC, thyroid-like follicular RCC, tubulocystic RCC and hybrid oncocytic/chromophobe tumors （HOCT）. In current review, we will collect available literature of these newly-described RCCs, analyze their clinical pathologic characteristics, discuss their morphologic and immunohistologic features, and finally summarize their molecular and genetic evidences. We expect this review would be beneficial for the understanding of RCCs, and eventually promote clinical management strategies.

关键词： Renal cell carcinoma （RCC） renal tumor Immunohistochemistry molecular genetics

Mycoplasma leachii causes bovine mastitis: Evidence from clinical symptoms,histopathology and Immunohistochemistry

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Journal of Integrative Agriculture 2019年第1期18卷 160-168页

作者： CHANG Ji-tao YU De-bin LIANG Jian-bin CHEN Jia WANG Jian-fa WANG Fang JIANG Zhi-gang HE Xi-jun WU Rui YU Li Division of Livestock Infectious Diseases/State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural Sciences College of Animal Science and Veterinary Medicine Heilongjiang Bayi Agricultural University (Daqing High-tech Industrial Development Zone) Pathology Laboratory/State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural Sciences

Twelve quarters of six lactating cows were inoculated with Mycoplasma leachii strain GN407 through intramammary ductal infusion; another 12 quarters were inoculated with heat-inactivated M.leachii culture medium as negative controls.Multidisciplinary procedures,including clinical assessment,etiology assessment,pathology and Immunohistochemistry(IHC),were used to elucidate the pathogenicity of M.leachii in bovine mastitis.From post-inoculation days(PIDs) 3 to 9,12 inoculated quarters developed mild to severe clinical mastitis and mammary tissue histopathological changes,including inflammatory cell infiltration and architectural destruction of mammary gland ducts.The M.leachii antigen was also detected by IHC in the mammary gland epithelial cells of the inoculated quarters as a weak signal on PID 6 and as a strong signal on PID 9.The control quarters also developed mild mastitis and histopathological changes on PID 9,and M.leachii was also detected by IHC.Throughout the experimental period,the quarters of the negative control cow were clinically and pathologically normal,and the M.leachii antigen was not detected.In conclusion,direct histological and immunohistochemical evidence confirmed that M.leachii causes clinical bovine mastitis through histopathological lesions induced by invasion of the pathogen into mammary gland cells and through inflammatory cell infiltration.

关键词： Mycoplasma leachii bovine mastitis pathogenicity histopathology Immunohistochemistry

Comparison of Immunohistochemistry with fluorescence in situ hybridization in determining the human epidermal growth factor receptor 2 status of breast cancer specimens： a multicenter study of 3149 Chinese patients

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Chinese Medical Journal 2014年第2期127卷 246-253页

作者： Han Xiaohong Shi Yuankai Ma Li Lyu Zheng Yang Hongying Yao Jiarui Li Jian Li Bo Qin Yan Department of Medical Oncology Beijing Key Laboratory of Clinical Study on Anti-cancer Molecular Targeted Drugs Cancer Institute/Hospital Chinese Academy of Medical Sciences & Peking Union Medical College Beijing 100021. China Department of Pathology Cancer Institute/Hospital Chinese Academy of Medical Sciences & Peking Union Medical College Beijing 100021. China

Background Accurate detection of human epidermal growth factor receptor 2 （HER2） expression and gene amplification is crucial for the application of HER2-specific therapy and for evaluating the response of patients with breast cancer.A uniform and standard procedure of Immunohistochemistry （IHC） and fluorescence in situ hybridization （FISH） needs to be established for evaluating the HER2 status in breast cancer tissues for the treatment of patients with real HER2-positive tumors.The present multicenter study was aimed to examine the HER2 status in breast cancer specimens from Chinese patients using both IHC and FISH methods.Methods A multicenter study was performed on the HER2 status in 3 149 breast cancer specimens from different ethnic populations and areas in China by IHC and FISH assays.The potential association of HER2 status with demographic and clinical characteristics was analyzed.Results The positive rates for HER2 over-expression and HER2 amplification were 23.3％ and 27.5％ in this study,respectively.The concordance between IHC and FISH was 71.2％（K=0.494,P ＜0.001）.Furthermore,72.9％ of specimens with IHC 2＋ were negative to FISH.The discordance rates among laboratories were from 5％ to 28％ for IHC and 1％ to 16％ for FISH.HER2 amplification was associated significantly with advanced tumor stage （Ⅲ or Ⅳ,P=0.002）,large tumor size （＞5 cm,P=0.002）,moderate and poor histological grades （P ＜0.0001）,post-menopause （P ＜0.0001）,ER-PR-（P=0.002）,and having ＞4 lymph nodes affected （P ＜0.0001） in this population.The positive rates of HER2 amplification in specimens from Man and Hui Chinese were significantly higher than that in other Chinese populations.There are slightly higher positive rates of HER2 expression and amplification in Chinese patients with breast cancer.Conclusion These findings may provide new insights into understanding the epidemiological features of HER2 expression and amplification,and may be valuable for clinical practice.

关键词： human epidermal growth factor receptor 2 fluorescent in situ hybridization Immunohistochemistry breast cancer

Can pathologists reliably establish ampulla of Vater carcinoma histologic subtype on H&E alone?Concordance of subtype and comparison to Immunohistochemistry-established subtype

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Journal of Pancreatology 2019年第4期2卷 152-157页

作者： Tanager Kevin S Li Yueying Alpert Lindsay Setia Namrata Weber Christopher Hart John Xiao Shu-Yuan Department of Pathology The University of Chicago MedicineChicagoIL Department of Pathology Wuhan University Zhongnan Hospital Wuhan University Center for Pathology and Molecular Diagnostics WuhanHubeiChina

Histologic subtyping of ampulla of Vater carcinoma(ampullary carcinoma,AC)may guide choice of adjuvant chemotherapy for advanced disease,as intestinal(INT)subtype may have a better treatment response and survival as compared to the pancreatobiliary(PB)and mixed(MIX)subtypes.However,ambiguous morphologic features can result in inconsistent subtyping among pathologists.Recently Immunohistochemistry(IHC)for cytokeratin(CK)7,CK20,CDX2,and the Mucin(MUC)family proteins had been used to aid in the subtyping.It is important to determine if accurate subtyping can be achieved based on hematoxylin and eosin(H&E)evaluation alone,or if IHC is required.In this study,5 gastrointestinal pathologists evaluated 34 cases of AC for histologic subtype(PB,INT,or MIX)based on H&E stain alone.IHC studies(CK7,CK20,and CDX2)were used to determine a gold standard subtyping for comparison(strong CK7 with weak CK20 and CDX2=PB;strong CK20 or CDX2 and weak CK7=INT;strong CK7 and CK20 or CDX2=MIX).Interobserver concordance(kappa)was calculated.Percent correctness of subtyping was calculated in relation to IHC-established subtype gold standard.Interobserver concordance for subtype was fair(kappa 0.31).Percent correctness for subtype against IHC-established subtype was 50%for a majority(3 or more)of pathologists.In all 11 cases with 4 or 5 pathologists being concordant,the subtype was also correctly identified.In addition,data on chemotherapy regimen and overall survival was gathered from the electronic medical record.Patients who were assigned a subtype at time of diagnosis tended to receive gemcitabine for PB subtype,and FOLFOX-based therapy for INT subtype,with some cases diagnosed as"not-otherwise specified"typically also receiving gemcitabine.Survival data were limited by loss to follow-up.In conclusion,despite 11 cases with strong concordance and correctness,overall subtyping concordance was only fair,and a majority of the 5 pathologists correctly identified the subtype in only 50%of cases.Thus,while some AC cases yield clear-cut subtypin

关键词： Ampulla of Vater Immunohistochemistry Immunophenotyping Prognosis Subtype

Diagnostic utility of IgG and IgM Immunohistochemistry in autoimmune liver disease

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World Journal of Gastroenterology 2010年第4期16卷 453-457页

作者： Roger Klein Moreira Frank Revetta Elizabeth Koehler Mary Kay Washington Department of Pathology Columbia University College of Physicians & SurgeonsNew YorkNY 10038United States Department of Pathology Vanderbilt UniversityNashvilleTN 37232United States Department of Biostatistics Vanderbilt UniversityNashvilleTN 37232United States

AIM:To assess the role of IgM and IgG Immunohistochemistry(IHC) in the evaluation of autoimmune liver conditions-autoimmune hepatitis(AIH),primary biliary cirrhosis(PBC),and primary sclerosing cholangitis(PSC).METHODS:Forty one biopsies from untreated patients diagnosed with autoimmune liver disease(AIH,n = 20;PBC,n = 13;PSC,n = 8) and fourteen biopsies of patients with chronic hepatitis C were selected.IgM and IgG-positive plasma cells were counted in each sample.RESULTS:A predominance of IgG-positive plasma cells was seen in AIH(90% of cases),PSC(75% of cases),and chronic hepatitis C(100% of cases),while IgMpositive plasma cells predominated in PBC(92.8% ofcases).The IgM /IgG ratio(< 1 or ≥ 1) accurately distinguished PBC from AIH in 90.9% of cases(sensitivity = 92.3%,specificity = 90%),and PBC from either AIH or PSC in 87.8% of cases(sensitivity = 92.3%,specificity = 85.7%).CONCLUSION:Plasmacytic infiltrates expressing predominantly IgM are characteristic of PBC,while other forms of liver disease analyzed in this study,including AIH,typically show an IgG-predominant plasma cell infiltrate.Our data indicate that IgM and IgG IHC may be a useful tool when PBC is a diagnostic consideration.

关键词： Autoimmune hepatitis Primary sclerosing cholangitis Primary biliary cirrhosis Immunoglobulin Immunohistochemistry

Androgen receptor expression in clinically localized prostate cancer： Immunohistochemistry study and literature review

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Asian Journal of Andrology 2008年第6期10卷 855-863页

作者： Yi-Qing Qiu Ivo Leuschner Peter Martin Braun Department of Urology the Second Affliated Hospital of Zhejiang University Hangzhou 310009 China Department of Urology and Pediatric Urology Department of Pathology Universitgitsklinikum Schleswig-Holstein Campus Kiel Kiel 24105 Germany

Aim： To evaluate androgen receptor （AR） expression in clinically localized prostate cancer （PCa）. Methods： Specimens were studied from 232 patients who underwent radical prostatectomy for clinically localized prostatic adenocarcinoma without neoadjuvant hormonal therapy or chemotherapy at our institution between November 2001 and June 2005. Immunohistochemical study was performed using an anti-human AR monoclonal antibody AR441. The mean AR density in the hot spots of different histological areas within the same sections were compared and the correlation of malignant epithelial AR density with clinicopathological parameters such as Gleason score, tumor, nodes and metastases （TNM） stage and pre-treatment prostate-specific antigen （PSA） value was assessed. Results： AR immunoreactivity was almost exclusively nuclear and was observed in tumor cells, non-neoplastic glandular epithelial cells and a proportion of peritumoral and interglandular stromal cells. Mean percentage of AR-positive epithelial cells was significantly higher in cancer tissues than that in normal prostate tissues （mean e SD, 90.0% ± 9.3% vs. 85.3% ±9.7%, P 〈 0.001）. The histological score yielded similar results. The percentage ofAR immunoreactive prostatic cancer nuclei and histological score were not correlated with existing parameters such as Gleason score, tumor, nodes and metastases stage and pre-treatment PSA value in this surgically treated cohort. Conclusion： The results of the present study suggest that there may be limited clinical use for determining AR expression （if evaluated in hot spots） in men with localized PCa.

关键词： androgen receptor prostate cancer Immunohistochemistry

Application of Immunohistochemistry technique in hydrobiological studies

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Aquaculture and Fisheries 2017年第3期2卷 140-144页

作者： Hui Zhang Wen Wen Jizhou Yan Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources Ministry of EducationShanghai Ocean UniversityShanghai 201306China Institute for Marine Biosystem and Neuroscience Department of BiologyCollege of Fisheries and Life SciencesShanghai Ocean UniversityShanghai 201306China

There is growing demand for biotechniques in oceanobiological and hydrobiological studies.Immunohistochemistry(IHC)combines anatomical,immunological and biochemical techniques to identify specific antigen-antibody interactions.Detecting the target antigen with labeled antibodies is a multi-step process that requires optimization at every level to maximize the signal detection.Here we use zebrafish samples and optimize each IHC step,particularly antigen retrieval,transgenic reporter double staining and background counterstaining.The resulting images with high signal intensity lead us to recommend the resulting protocols for experiments with fish.

关键词： Immunohistochemistry Protocol Transgenic fish Hydrobiology