To explore the significance of the application of standardized special staining technique in pathologic diagnosis of pulmonary fungal infectious diseases.Methods Final pathologic diagnosis of 104 cases pulmonary fungus infection disease in Beijing Chaoyang hospital from September 2011 to March 2016 were selected;HE staining only,HE staining combined with the traditional manual special staining method PAS and hexamine silver,and HE staining combined with automatic special staining PAS and hexamine-silver were used and compared.The two kinds of special staining technology were compared;the microscopic observation,analysis results (all the first staining results,not including the results of complex staining),the results on the basis of final pathologic diagnosis were also compared with the clinical preliminary diagnosis.Results Lung fungal infectious disease diagnosis rate and fungal classification rate,from low to high order consistence,showed that for the primary clinical diagnosis (29.8％ and 19.2％),HE staining (32.7％ and 32.7％),HE staining combined with traditional manual special staining method PAS and hexamine silver (90.4％ and 87.5％),and HE staining combined with automatic special staining PAS and hexamine-silver (98.1％ and 94.2％).The four methods were statistically significant on two aspects (P ＜ 0.01,P ＜ 0.01);the fourth method was significantly different from the first two (P ＜ 0.01,P ＜ 0.01).The fourth method was significantly different from the third kind of diagnosis rate (P ＜ 0.05),typing rate was no significant difference (P ＞ 0.05).But automatic special dyeing method of PAS and hexamine silver steps were more simple,with standardized chemical reagents,no artificial and environmental factors,short time-consuming,and less number of dropping-off and restaining of the section.Conclusion HE staining and its combination with automatic special staining of PAS and hexamine silver are much more standardized,and help to improve the diagnosis of pulmonary fungal infectious diseases and fungal classification rate.

Esophagus ; pathology ; Gastroenterology ; trends ; Humans ; Stomach ; pathology

Objective To investigate the effect of mycophenolic acid on the proliferation of smooth muscle cells (SMCs) in rats' pulmonary arterial at cellular level. Methods Growth curve, MTT tests, and flow cytometry were used to detect the number of proliferative cells, the A-value of living cells and the DNA content of the control and drugs groups respectively and the number of cells in G1-phase, S-phase, G2M- phase and the proliferation index were calculated. Results Compared with the control group, the number of living cells in the mycopbenolic acid groups (with the concentration of 1, 10, 100 μmol/L) decreased, except the lowest dosage group (100 nmol/L). The difference was statistically significant (P<0.01). The living cells measured by MTT dose-dependently reduced in the mycophenolic acid groups. In the mycophenolic acid groups, the fraction of living cells in the S-phase and G2M-phase decreased respectively while that in G1- phase increased, and the proliferation index decreased. All these responses presented with a dose-dependent manner. Conclusion Mycophenolic acid can effectively inhibit the proliferation of rats' pulmonary arterial SMCs. This process happens mainly in ONA synthesis phase, and is dose dependent. In addition, these effective concentrations are all in the available range for clinical application.

Objective To investigate the correlation between the clinical pathological factors and the expression of EGFR, HER2 and HER3 in non-small cell lung cancer tissues. Methods The expression of EGFR, HER2 and HER3 in non-small cell lung cancer tissues was detected by MaxVision immunohistochemical. The relationship between the protein expression and the clinical pathologic characteristics was studied. The correlation between the protein expression and the survival was also retrospectively analyzed. Results Increased expression of EGFR, HER2 and HER3 was detected in the NSCLC tissues and their rates were 45.9 % (73/156), 30.8 % (49/156) and 21.4 % (37/156), respectively. There were significant associations between the expression of these proteins and lymph node metastasis, tumor differentiation degree and TNM staging. Conclusions Overexpression of EGFR, HER2, HER3 or all three proteins is a predictor of poor prognosis. Moreover, combined detection of all 3 markers may work better than the individual detection in the prediction of the prognosis to guide the clinical treatment.

Objective To establish a zebrafish model of thrombosis induced by three kinds of inducers and observe the anti?thrombotic effect of a Chinese traditional medicine, Guanxinning tablet ( GXN) . Methods The zebrafish models of thrombosis was induced by using 1?5μmol/L phenyl hydrazine, 80μmol/L arachidonic acid and 5 mg/L ponatinib, re?spectively, and were treated with various concentration of GXN, clopidogrel or asprin. The thrombus in the tail vein was observed under microscope, Erythrocytes in the zebrafish heart were stained with o?dianisidine and the erythrocyte staining intensity was assessed with a NIS?Elements DTM image analyzer, and the anti?thrombolic effect of GXN was calculated. Results Venous thrombus was significantly increased and the staining intensities of erythrocytes in the heart were signifi?cantly decreased after induction by phenyl hydrazine, arachidonic acid or Ponatinib ( P <0?001 ) , respectively. At the same time, GXN showed an incresing anti?thrombolic effect in the zebrafish models (P<0?001) in a dose?effect manner, with a IC50 of GXN of 44?32 mg/L,138?5 mg/L and 459?5 mg/L, respectively. Conclusions The zebrafish models of thrombosis are successfully established by phenyl hydrazine, arachidonic acid or Ponatinib, respectively, by different for?mation mechanisms. GXN has been shown to have an anti?thrombosis effect, probably, by multiple target effects.

Objective To explore rules of changes in velocity of ultrasound wave transmission at the tibia and radius by a quantitative bone mineral density ultrasound scanner in examination of early incipient rickets in infants and its significance in clinical diagnosis.Methods One hundred and fifty-seven infants who visited child health-care clinic of the Capital Institute of Pediatrics,Beijing during May 2004 to December 2007 and clinically diagnosed as rickets according to the Criteria of Diagnosis for Rickets in Infants and Young Children formulated in 1986 by the Ministry of Health,as well as 124 normal healthy infants as controls,were enrolled in the study and followed-up for three months.Velocity of ultrasound wave transmission at the tibia and radius in all the infants were measured by a bone mineral density ultrasound scanner (Sunlight Omnisense 7000R made in Israel).Results One hundred and fifty-seven infants were clinically diagnosed as rickets,127 of them undergone with carpal plane roentgenography and 90 of the 127 with positive change in bone x-ray imaging and 37 without it.Velocity of ultrasound wave transmission at the tibia and radius measured by z-score was significantly lower in infants with clinically diagnosed rickets than that in healthy controls (Z-values of-10.411 and-10.399 at the tibia and-5.646 and-5.517 at the radius,respectively,P = 0.000 with Mann-Whitney test).Velocity of ultrasound wave transmission at the tibia and radius measured by z-score was lower in those with positive change in x-my imaging than that in those without it,but not reaching a level of statistically significant difference.Velocity of ultrasound wave transmission and z-score correlated positively with change in bone x-ray imaging,respectively with Spearman coefficients of correlation of 0.581 and 0.677 for tibia,0.316 and 0.467 for radius (P = 0.000).Receiver operating characteristic (ROC) curve was drawn from those with rickets and positive left carpal plane roetgenograph.Area under curve of ROC for z-score of velocity of ultrasound wave transmission at the tibia was 0.812 and 0.799 (95% CI 0.758-0.856 and 0.742-0.855,P =0.000),respectively.Area under ROC curve of z-score of velocity of ultrasound wave transmission at the radius was 0.715 and 0.697 (95% CI 0.650-0.780,0.631-0.764,P =0.000),respectively.Cut-off value of z-score was-205--1.95 at the tibia,according to the largest sensitivity and the least false-positivity,with sensibility of 0.8 to 0.9 and specificity of 0.733 to 0.702.As its sensitivity at the radius was more than 0.7,its specificity was lower than 0.524.Conclusions Quantitative ultrasound scanning can be used to detect insufficient bone mineral density in infants and their early skeletal change by rickets but without change in bone x-ray imaging.Velocity of ultrasound wave transmission at the tibia can be used as an indicator to predict early rickets in infants rather than its severity.

Objective:To investigate the correlation of CD8 positive tumor-infiltrating lymphocytes (CD8 + TIL) density and programmed-death receptor ligand 1 (PD-L1) expression in rectal cancer with clinicopathological characteristics and prognosis of patients after neoadjuvant chemoradiotherapy. Methods:The clinicopathological data of 166 patients with locally advanced rectal cancer (LARC) who received neoadjuvant therapy before surgery in the Beijing Chao-Yang Hospital, Capital Medical University from January 2015 to December 2018 were retrospectively analyzed. CD8 + TIL density and PD-L1 expression were detected by using immunohistochemistry. The correlation of CD8 + TIL density and PD-L1 expression with clinicopathological characteristics of patients after neoadjuvant chemoradiotherapy was analyzed. Kaplan-Meier method was used to analyze the disease-free survival (DFS) and Cox regression risk model was used to make univariate and multivariate analysis of the influencing factors for DFS. Results:Among 166 LARC patients, 81 cases (48.8%) had high density of CD8 + TIL, 85 cases (51.2%) had low density of CD8 + TIL; 63 cases (38.0%) had PD-L1 expression, and 103 cases (62.0%) had non-expression of CD8 + TIL. The expression rate of PD-L1 in CD8 + TIL high density group was higher than that in CD8 + TIL low density group [50.6% (41/81) vs. 25.9%(22/85), χ2 = 10.78, P < 0.001]. According to the density of CD8 + TIL and PD-L1 expression, immunophenotype was divided among 4 groups; the 3-year DFS rate of the CD8 + TIL high density /PD-L1 expression group was 87.1%, which was higher than that of the other groups (CD8 + TIL low density /PD-L1 expression group was 72.8%, CD8 + TIL high density /PD-L1 non-expression group was 67.0%, CD8 + TIL low density /PD-L1 non-expression group was 64.3%), and the difference was statistically significant ( P < 0.05). Univariate analysis showed that tumor differentiation degree, TNM stage, CD8 + TIL density, PD-L1 expression and CD8 + TIL density /PD-L1 expression were correlated with the DFS of patients (all P < 0.05). Multivariate analysis results showed that CD8 + TIL high density /PD-L1 expression was an independent protective factor for DFS ( HR = 0.049, 95% CI 0.005-0.497, P = 0.011), while TNM stage 3 was an independent risk factor for DFS ( HR = 2.752,95% CI 1.300-5.825, P = 0.008). Conclusions:In LARC after neoadjuvant therapy, CD8 + TIL density is positively correlated with the expression of PD-L1, and the high density of CD8 + TIL/PD-L1 expression is an independent influencing factor for good prognosis, suggesting that these patients may benefit from the immunotherapy.

OBJECTIVETo study the difference of microRNA (miRNA) expression between two groups of early stage (pT1N0) esophageal squamous cell carcinoma (ESCC) patients who had different outcome and the prognostic significance of different miRNA in metastatic of early ESCC, and to identify useful prognostic markers in the selection of appropriate treatment for early ESCC patients.

METHODSTaqMan human miRNA arrays and bioinformatics were used to detect and analyze the expression profiles of miRNAs in the two groups, and RT-PCR was used to verify the differences in miRNA expression.

RESULTSThe miRNA arrays revealed a total of 41 markedly changed miRNAs in the survival group compared with the death group. Bioinformatics analysis, prediction and significant function analyses of targeted genes and pathway analysis identified that miR-27a, miR-143 and miR-886-5p levels were increased or decreased by seven-folds or more. The enriched target genes were GRB2, SOS1, MAPK1, EGFR, CBL, SPRY2, RPS6KA5, IGF1R, NGFR, MAPK14 and CREB1. These genes were significantly related to the following signaling pathways, i.e.Sprouty regulation of tyrosine kinase signals pathway, Erk1/Erk2 Mapk signaling pathway and transcription factor CREB and its extracellular signals.

CONCLUSIONSmiR-27a, miR-886-5p, and miR-143 may be potential prognostic markers of metastasis for early ESCC. The detection of these miRNAs plays a directive role for the treatment options of early ESCC. The regulation of targeted genes and mechanism remain to be further studied.

Biomarkers, Tumor ; genetics ; metabolism ; Carcinoma, Squamous Cell ; genetics ; pathology ; surgery ; Esophageal Neoplasms ; genetics ; pathology ; surgery ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; MicroRNAs ; metabolism ; Neoplasm Metastasis ; Neoplasm Staging ; Signal Transduction

Purpose To investigate the clinicopathological and histological features of non-small cell lung cancer(NSCLC)patients with high expression of PD-L1 and positive driver muta-tion.Methods The clinical data of 141 patients with PD-L1 high expression and driver mutation-positive NSCLC were col-lected.Immunohistochemical methods,ARMS-PCR,and next-generation sequencing(NGS)were used to detect PD-L1 ex-pression and driver gene mutations.The clinicopathological fea-tures were analyzed and the related literatures were reviewed.Results There were 141 cases NSCLC patients with high ex-pression of PD-L1 in tumor cells,of which 57 cases were≥50％,＜60％;≥60％,＜70％in 18 cases;≥70％,＜80％in 35 cases;≥80％in 31 cases.Among 141 cases NSCLC patients with high PD-L1 expression,53 cases(37.6％)had driver gene mutations,including 4 cases BRAF 15 exon mutations,9 cases MET-associated mutations,17 cases EGFR-associated mutations,16 cases KRAS 2 exon mutations,4 cases EML4-ALK fusion mutations,and 3 cases other rare mu-tations.The high expression of PD-L1 and the occurrence of driver gene mutation were related to the gender,smoking history and pathological type of patients(P＜0.05).MET-related mu-tations and KRAS 2 exon mutations were more common in males than in females.All BRAF 15 exon mutations were female.The mean percentage of PD-L1 expression was highest in patients with MET mutation,KRAS 2 exon mutation,and 3 cases rare mutations.In 33 cases with BRAF 15 exon mutation,MET am-plification or mutation,EGFR-related mutation,and 3 cases oth-er rare mutations,PD-L1 was highly expressed in solid,glandu-lar,and micropapillary tumor cells.In 20 cases with KRAS 2 exon mutation and EML4-ALK fusion mutation,PD-L1 was highly expressed in solid nested tumor cells.Conclusion In NSCLC,high expression of PD-L1 and positive driver gene mu-tation are negatively correlated with the degree of tumor differen-tiation.In the poorly differentiated surgical specimens of lung adenocarcinoma,solid,micropapillary,or glandular tubular tumor tissues should be selected as far as possible for PD-L1 ex-pression and driver gene mutation detection.

The pursuit of minimally invasive biopsy and targeted treatment as well as technical progress have boosted the extensive clinical usages of fine needle puncture cell/tissue acquisition technology in recent years. How to rationally use the limited fine needle puncture materials which are usually minimal and fragmented, while making pathological diagnosis, accomplish the auxiliary tests of immune phenotype, molecular features and other tests required by the clinician, and comprehensively improve the diagnostic benefits of fine needle puncture material is a big challenge facing the pathology community. Up to now, there has been no successful experience that can be learned from abroad. Some suggestions for the further improvement of diagnostic efficacy of fine needle puncture specimens from the aspects of sub-specialty cooperation and comprehensive utilization of the materials are put forward based on the progress trend in pathology and the current situation in China.
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