To explore the automated immunostainer screening anaplastic lymphoma kinase (ALK) gene fusion non-small cell lung cancer (NSCLC) and clinicopathological characteristics of the molecular subtype lung cancers. Methods Five hundred and sixty-six cases of NSCLC were collected over a 16 month period. The test for ALK was performed by Ventana automated immunostainer with anti-ALK D5F3. The histological features, treatment and outcome of patients were assessed. Results Thirty-eight cases (6.7%, 38/566) of NSCLC showed ALK gene fusion. The frequency of ALK gene fusion was higher in male (7.1%, 25/350) than that in female (6.0%, 13/216) patients, but not achieving statistical significance (chi2 = 0.270, P = 0.604). ALK + NSCLC was more significantly more frequent in patients < or = 60 years (9.9%, 28/282) than >60 years (3.5% , 10/284) of age. Histologically, the ALK + NSCLCs were mostly adenocarcinoma (81.6%, 31/38) , among which eighteen cases were solid predominant subtype with mucin production; nine cases were acinar predominant subtype; one case was papillary predominant subtype and three cases were invasive mucinous adenocarcinoma. The ALK + non-adenocarcinoma included three cases of squamous cell carcinoma, three cases of adenosquamous carcinoma and one case of pleomorphic carcinoma. Among the ALK + NSCLC patients, the number of non/light cigarette smokers (86. 8% , 33/38) was more than that of heavy smokers. Twenty-nine cases were stages III and IV; twenty-nine cases showed lymph node metastasis; twenty cases showed metastases mostly to brain and bone; and one case showed EGFR gene mutation coexisting with ALK gene fusion. Twelve of fifteen patients received crizotinib therapy and remained stable. Conclusions NSCLC with ALK gene rearrangement shows distinctive clinical and histological features. Ventana-IHC may he a feasible and valid technique for detection of ALK rearrangement in NSCLC.
Adenocarcinoma ; genetics ; pathology ; Carcinoma, Adenosquamous ; genetics ; pathology ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; Carcinoma, Squamous Cell ; genetics ; pathology ; Female ; Gene Fusion ; Gene Rearrangement ; Humans ; Lung Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Protein Kinase Inhibitors ; therapeutic use ; Pyrazoles ; therapeutic use ; Pyridines ; therapeutic use ; Receptor Protein-Tyrosine Kinases ; genetics ; Sex Factors

Objective: To investigate the clinicopathological, and molecular characteristics of myoepithelial tumors (MTs) of salivary glands. Methods: A total of 37 MTs cases including 13 malignant epithelial tumors (MMTs) and 24 benign epithelial tumors (BMTs) of salivary glands were identified from the archives of the Department of Pathology, General Hospital of Eastern Theater Command, dating from 2006 to 2016. Clinical features, histological patterns, immunohistochemical characteristics and status of EWSR1 gene rearrangement by fluorescence in situ hybridization (FISH) analysis were reviewed in all cases. Results: Clinically, 37 MTs cases mainly occurred in the parotid glands, when most of the patients presented with painless masses. Of the 13 MMTs cases, male to female ratio was 7∶6, and the median age was 62 years old. Of the 24 BMTs cases, male to female ratio was 5∶7, and the median age was 54 years old. Immunohistochemically, 37 MTs cases were positive for CKpan, and at least one myoepithelial marker. Twenty six of 37 MTs cases were analyzable for the EWSR1 gene break by FISH. Based on the previous evaluation criterion, the EWSR1 translocation was detected in 4 cases of 11 MMTs, and 4 cases of 15 BMTs. According to the main histological composition of tumor cells, 4 EWSR1-positive MMTs covered 2 clear-cell cases and 2 epithelioid-cell cases, when 4 EWSR1-positive BMTs covered 2 clear-cell cases, 1 plasmacytoid-cell case, and 1 spindle-cell case. Conclusions Males and females are affected equally. MTs express immunoreactivity for CKpan, and at least one myoepithelial marker. The EWSR1 rearrangement is present in a subset of MTs, with variable morphological characteristics, and has no statistical significance on clinical behavior.

Objective: To compare amplification refractory mutation system(ARMS) and droplet digital PCR (ddPCR) in the detection of epidermal growth factor receptor (EGFR) gene mutations in patients with non-small cell lung cancer (NSCLC), and to investigate the clinical value of ddPCR. Methods: A total of 79 specimens of NSCLC, including 22 cases of cell block, 18 cases of surgical specimens, 12 cases of biopsy specimens and 27 cases of plasma samples, were analyzed for the mutation status of EGFR gene by ARMS and droplet digital PCR method. Results: In 18 cases of surgical specimens and 12 cases of biopsy specimens, the detection results by the two methods were identical with positive rates of 9/18 and 5/12, respectively. In 22 cases of effusion cell blocks, ARMS detected 19-del and L858R of EGFR gene in two cases, in which droplet digital PCR detected 19-del+ T790M mutations in one case and L858R+ T790M mutation in another. L858R mutation was detected by droplet digital PCR in one case but ARMS assay was negative. The remaining 19 cases were consistent by the two methods. In blood samples, the positive rate was 33.3%(9/27) by ARMS and 37.0%(10/27) by droplet digital PCR. Two cases showed L858R and 19-del+ T790M mutation by droplet digital PCR but ARMS assay detected only 19-del. The remaining 25 cases were consistent by the two methods. Conclusion Droplet digital PCR method is more sensitive and accurate than ARMS for the detection of EGFR mutations in pleural fluid and blood samples, can be used in clinical test.

Objective:To establish an analytical method for the simultaneous determination of five fat-soluble vitamins in serum using isotope dilution ultra high performance liquid chromatography-tandem mass spectrometry (ID-UPLC-MSMS).Methods:Fat-soluble vitamins were obtained from serum samples which collected from Shanghai Tenth People′s Hospital between April 2019 and August 2019 by the extraction method, and were detected by ID-UPLC-MSMS. The performance of the method was verified by referring to the relevant documents of the Clinical and Laboratory Standards Institute (CLSI).Results:The ID-UPLC-MSMS method for the rapid detection of various fat-soluble vitamins in serum was proposed and successfully verified. The linear range of the method: vitamin A: 25-2 500 μg/L, 25(OH)D 2: 2-200 μg/L, 25(OH)D 3: 2-200 μg/L, vitamin E: 0.25-50 mg/L, vitamin K1: 0.1-20 μg/L. The intra- and inter-assay precision standard deviations of the five analytes were within ± 15%, and the accuracy of the test results of the 25(OH)D 2 and 25(OH)D 3 standards was 96.44%-102.37%. Conclusion:The performance of ID-UPLC-MSMS method for the simultaneous determination of five fat-soluble vitamins is satisying, and the result is accurate and reliable, which suggested it can be used for the clinical sample.

Objective To evaluate the clinical efficacy of percutaneous transluminal angioplasty (PTA) combined with stent implantation in the treatment of transplant renal artery stenosis (TRAS) after renal transplantation. Methods Clinical data of 21 patients with TRAS after renal transplantation undergoing PTA combined with stent implantation were retrospectively analyzed. The incidence of TRAS in renal transplant recipients was summarized. The changes of relevant indexes in patients with TRAS were statistically compared before and after interventional treatment. Clinical prognosis of patients with TRAS was evaluated. Results The incidence of TRAS in renal transplant recipients was 4.1%(21/507). TRAS was diagnosed at postoperative 5 (4, 7) months, and 67% (14/21) of patients developed TRAS within postoperative 6 months. Compared with the values before interventional therapy, the serum creatinine level, systolic and diastolic blood pressure and peak flow velocity of transplant renal artery of patients with TRAS were significantly decreased, and the estimated glomerular filtration rate (eGFR) and interlobar arterial resistance index were significantly increased at 1 week and 1 month after interventional therapy (all P < 0.05). During postoperative follow-up after PTA combined with stent implantation, 1 patient suffered re-stenosis of the transplant renal artery, which was improved after simple balloon dilatation. One patient developed pseudoaneurysm formation at the puncture site of the right femoral artery. One patient presented with renal atrophy and loss of function due to atresia of the transplant renal artery. All the remaining 18 patients were well recovered after surgery. Conclusions PTA combined with stent implantation is the optimal treatment of TRAS after renal transplantation, which can significantly improve the function of transplant kidney and considerably prolong the survival time of transplant kidney.

OBJECTIVETo study the expression, clinicopathologic correlation and prognostic significance of caveolin-1 in lung adenocarcinomas(LAC).

METHODSImmunohistochemical study (EnVision method) for caveolin-1 and TTF-1 was carried out in 185 cases of LAC encountered during the period from 2005 to 2010. The correlation between caveolin-1 expression and various clinicopathologic parameters was analyzed statistically.

RESULTSThe rate of caveolin-1 expression in the 185 cases of LAC was 26.5% (49/185) and significantly lower than that in normal lung tissue (P<0.01). There was also higher rate of caveolin-1 expression in male patients (P=0.004), smokers (P=0.006), tumors larger than 3.5 cm (P=0.048), predominantly solid tumor subtype (P=0.025), high tumor grade (P=0.044), tumors with vascular invasion (P=0.019), lymph node metastasis (P=0.030), recurrence (P=0.021) and high clinical stage (P=0.027). The expression level of caveolin-1 in TTF1-negative cases was significantly higher than that in TTF1-positive cases and caveolin-1 expression also negatively correlated with TTF-1 expression in LAC (r=-0.154, P=0.037). The five-year overall survival rate of patients with caveolin-1 positive tumors was lower than that in caveolin-1 negative group (P<0.01).Univariate analysis indicated the expression level of caveolin-1 and TTF-1 (P<0.01), histologic subtype (P=0.002), tumor grade (P=0.002), tumor size (P=0.009), vascular invasion (P=0.019), lymph node metastasis (P=0.018), recurrence (P=0.032) and clinical stage (P=0.024) correlated with the survival of patients with LAC. COX multivariate analysis revealed that LAC with caveolin-1 positive expression, TTF-1 negative expression and high tumor grade carried a significantly unfavorable prognosis.

CONCLUSIONCaveolin-1 expression correlates with histologic subtype, tumor grade, invasiveness and metastatic potential of LAC. The detection of caveolin-1 in LAC is helpful in predicting prognosis.LAC with caveolin-1 expression carries a poor prognosis.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adenocarcinoma, Papillary ; metabolism ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Acinar Cell ; metabolism ; pathology ; surgery ; Caveolin 1 ; metabolism ; DNA-Binding Proteins ; metabolism ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; metabolism ; pathology ; surgery ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis ; Survival Rate ; Transcription Factors ; Tumor Burden

OBJECTIVETo study the clinicopathological features, differential diagnosis and prognosis of clear cell papillary renal cell carcinoma (CCPRCC).

METHODSThe histological, immunohistochemical, and molecular features were studied in 11 cases and follow-up data were also analyzed.

RESULTSThere were a total of 3 females and 8 males. The age of patients were ranged from 33 to 72 years(mean age 52.5 years). The diameters of tumors varied from 1cm to 4 cm. Histologically, papillary and cystic architecture were present at least focally in all tumors. The papillae were covered by small to medium-sized cuboidal cells with abundant clear cytoplasm and often showed extensive secondary branching, which were often folded and densely packed, resulting in a solid appearance. The nuclei were round and uniform in shape; nucleoli were not prominent (Fuhrman grade 1 or 2). Neither mitotic figures nor necrosis was present. All 11 cases exhibited moderate to strong positivity for CK7, CA9, vimentin, and HIF-1α, coupled with negative reactions for CD10, P504S, and TFE3. Ksp-cadherin was positively expressed in 8 cases.VHL gene mutations were not found in all 11 cases. Losses of chromosomes 3 (monoploid chromosome 3) was detected in 3 cases.

CONCLUSIONSCCPRCC is uncommon and seemed to be an indolent tumor. The differential diagnosis should be included tumors, which harbor clear cell and papillary structure including clear cell renal cell carcinoma, papillary renal cell carcinoma, Xp11 translocation renal cell carcinoma, and CCPRCC. Immunohistochemical and molecular analysis may be help for its diagnosis.

Adult ; Aged ; Carcinoma, Renal Cell ; chemistry ; genetics ; pathology ; ultrastructure ; Chromosomes, Human, Pair 3 ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; Kidney Neoplasms ; chemistry ; genetics ; pathology ; ultrastructure ; Male ; Middle Aged ; Mutation ; Prognosis ; Racemases and Epimerases ; analysis ; Translocation, Genetic ; Tumor Burden

OBJECTIVETo study the immunophenotype and molecular genetics of epithelioid sarcoma (ES), INI1 expression and its role in differential diagnosis.

METHODSTwenty cases of ES were retrieved from the archival files and selected for immunohistochemical study, DNA sequencing and fluorescence in-situ hybridization. The clinical and pathologic features were also reviewed.

RESULTSThe age of patients ranged from 16 to 75 years (mean = 40.2 years). The median age of patients in classic ES and proximal-type was 37.9 years and 42.0 years, respectively. The male-to-female ratio was 1.2: 1.0. Classic ES mostly occurred in the extremities while proximal-type ES often affected the perineum and external genitalia and trunk. Histologically, granuloma-like structures, consisting of aggregates of epithelioid and spindly tumor cells with central necrosis, were observed in classic ES. The epithelioid tumor cells contained abundant eosinophilic cytoplasm, merged with spindly cells at the periphery and admixed with collagen fibers. In proximal-type ES, the tumor cells showed prominent epithelioid and/or rhabdoid features, had marked cytologic atypia and grew in multinodular or diffuse patterns. In 2 cases of proximal-type ES studied, the "rhabdoid" tumor cells demonstrated a diffuse sheet-like growth pattern, mimicking malignant rhabdoid tumor. Immunohistochemical study showed that vimentin was positive in all cases. Pan-cytokeratin, CK8, CK7, epithelial membrane antigen and CD34 were expressed in 16, 15, 1, 18 and 13 cases, respectively. The staining for S-100 protein was focal and weak in 5 cases. None of the cases studied expressed CD31 and HMB45. Loss of INI1 was demonstrated in 10 of the 13 classic ES cases and 5 of the 7 proximal-type ES cases. The mutation of INI1 gene was detected in 1 of the 6 cases. Deletion of INI1 gene including heterozygous deletion, homozygous deletion and haploid was observed in 8 of the 11 cases.

CONCLUSIONSOwing to the histologic heterogeneity, pitfalls in diagnosis of ES sometimes are encountered. INI1 is lost in most cases of ES. Immunohistochemical study, including staining for INI1, provides useful clues in pathologic diagnosis. Instead of INI1 mutation, inactivation of INI1 gene related deletion is not uncommon.

Adolescent ; Adult ; Aged ; Antigens, CD34 ; metabolism ; Chromosomal Proteins, Non-Histone ; genetics ; DNA-Binding Proteins ; genetics ; Female ; Gene Deletion ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Keratins ; metabolism ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Mutation ; Rhabdoid Tumor ; genetics ; SMARCB1 Protein ; Sarcoma ; genetics ; Transcription Factors ; genetics ; Young Adult

OBJECTIVETo investigate the clinicopathologic features, immunohistochemic phenotypes and genetic alterations of extrapulmonary inflammatory myofibroblastic tumor (IMT) and the correlation with prognosis.

METHODSThirty cases of IMT with follow-up were analyzed morphologically and immunohistochemically. ALK FISH was also performed to determine the ALK gene status.

RESULTSPatients ranged in age from 12 to 73 years (mean 43.4 years). The male-to-female ratio was 1.0: 1.1. The tumors were located in various anatomical sites including gastrointestinal tract, liver, spleen, kidney, pelvic, retroperitoneum, mediastinum etc. Histologically, the majority of cases were composed of spindled fibroblastic and myofibroblastic cells accompanied by an inflammatory infiltrate of plasma cells, lymphocytes, and eosinophils. Most cases with aggressive behavior had features including prominent nucleoli and edematous myxoid background. Lymphohistiocytic reactions were usually absent. Some cases showed multinucleation, nuclear pleomorphism and mitoses. One case demonstrated epithelioid morphology with round-to-epithelioid cells. The immunohistochemical study showed vimentin, SMA, CK, desmin, and ALK were expressed in 100% (30/30), 70% (21/30), 13% (4/30), 27% (8/30), and 27% (8/30) of IMT, respectively. Diffuse cytoplasmic ALK staining was detected in seven cases. One case (containing round-to-epithelioid cells) demonstrated ALK nuclear membrane staining, coupled with positive reaction for CD30 and negative reaction for LCA. EMA, CD34, CD117 and S-100 protein, and MyoD1 were negative for all cases. Six ALK protein positive cases harbored ALK gene rearrangement, but not the remaining 22 cases. Follow-up data were available in 21 patients. After initial resection, 14 patients were alive with no evidence of disease, while 4 patients were alive with tumor recurrence and 3 patients died of the disease.

CONCLUSIONSMost IMT with aggressive behavior have features including prominent nucleoli, edematous myxoid background, and positive expression of ALK. Lymphohistiocytic reaction is usually absent. ALK may be a potential novel therapeutic target for IMT.

Adolescent ; Adult ; Aged ; Child ; Female ; Fibroblasts ; pathology ; Granuloma, Plasma Cell ; genetics ; pathology ; Humans ; Inflammation ; pathology ; Male ; Middle Aged ; Myofibroblasts ; pathology ; Prognosis ; Receptor Protein-Tyrosine Kinases ; genetics ; Young Adult

OBJECTIVETo study the clinicopathologic features, immunophenotype, differential diagnosis and prognosis of renal cell carcinoma (RCC) associated with t(6;11)(p21.2;q13)/MALAT1-TFEB gene fusion.

METHODSA total of 9 cases of such rare tumor were selected for clinicopathologic, immunohistochemical and molecular analysis, with review of literature.

RESULTSThe age of the patients ranged from 21 to 42 years (mean=31.3 years). The patients included four men and five women. Histologically, 4 of the 9 cases studied showed classic morphologic features of TFEB RCC, with hyaline material, pigments and psammoma bodies frequently identified. The remaining 5 cases demonstrated uncommon morphology, mimicking perivascular epithelioid cell neoplasm, clear cell RCC, chromophobe RCC or papillary RCC. Immunohistochemical study showed that TFEB and vimentin were positive in all cases. Most of the tumors studied also expressed Ksp-cadherin, E-cadherin, CD117, HMB45, Melan A and Cathepsin K. CKpan showed immunostaining in only 1 case. The staining for TFE3, CD10 and CK7 were all negative. TFEB gene rearrangement was detected in all the 9 cases studied using fluorescence in-situ hybridization. MALAT1-TFEB fusion gene was identified in 2 cases by polymerase chain reaction and direct sequencing. TFEB RCC seemed to be an indolent tumor. During a mean follow-up of 31 months, none developed tumor recurrence, progression, or metastasis.

CONCLUSIONSTFEB fusion-associated RCC is a rare neoplasm, tends to occur in young age group and carries an indolent behavior. Diagnosis relies on clinicopathologic findings and immunohistochemical analysis. TFEB break-apart FISH assay is a reliable tool in confirming the diagnosis.

Adult ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; genetics ; Carcinoma, Renal Cell ; genetics ; pathology ; Chromosomes, Human, Pair 11 ; Chromosomes, Human, Pair 6 ; Diagnosis, Differential ; Female ; Gene Fusion ; Gene Rearrangement ; Genes, Neoplasm ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Kidney Neoplasms ; genetics ; pathology ; Male ; Prognosis ; RNA, Long Noncoding ; genetics ; Translocation, Genetic ; Young Adult