Diagnostic accuracy of a real-time PCR assay for detection of Helicobacter pylori and resistance to clarithromycin and levofloxacin directly from stool.

OBJECTIVE: The non-invasive detection of Helicobacter pylori (H. pylori) and its resistance to clarithromycin and levofloxacin significantly improves the management of infected patients by enabling tailored eradication treatments without the need for endoscopic procedures. This study aimed to assess the effectiveness of real-time PCR (RT-PCR) assays in identifying H. pylori infection and antibiotic resistance in stool and gastric biopsy specimens. PATIENTS AND METHODS: Stool and gastric biopsy samples were collected from patients within three days of post-hospitalization. A total of 115 samples were analyzed for H. pylori infection, and an additional 115 samples were evaluated for resistance to clarithromycin and levofloxacin using an RT-PCR-based molecular test. Statistical analyses were performed using (SPSS 26.0 IBM Corp., Armonk, NY, USA). RESULTS: Among 115 patients (53 males, average age 50.8±13.2 years), H. pylori was detected in 93.1% of stool samples and 93.9% of gastric biopsies. The RT-PCR assay demonstrated a sensitivity of 99.1% and a specificity of 100%, with an overall diagnostic accuracy of 99.1%. Clarithromycin resistance was found in 37.3% of stool and 46.9% of gastric biopsy specimens, with the assay showing 79.6% sensitivity and 98.4% specificity. Levofloxacin resistance was identified in 32.1% of stool samples and 31.3% of gastric biopsies, with 86.3% sensitivity and 91.1% specificity of the molecular test. CONCLUSIONS: The RT-PCR-based detection of H. pylori and its resistance to clarithromycin and levofloxacin in stool samples represents a promising approach to enhance eradication therapy outcomes, potentially improving treatment efficacy. Chictr.org.cn: ChiCTR2300070267.

[SRF-rearranged cellular perivascular myoid tumor: a clinicopathological analysis of two cases].

Objective: To investigate the clinicopathological features, immunophenotype, diagnosis and differential diagnosis of SRF-rearranged cellular perivascular myoid tumor. Methods: Two cases of SRF-rearranged cellular perivascular myoid tumor diagnosed in the Department of Pathology, Fudan University Shanghai Cancer Center from October 2021 to March 2022 were collected. Immunohistochemical staining, fluorescence in-situ hybridization (FISH) and next-generation sequencing (NGS) were performed, and the literature was reviewed. Results: Case 1, a 3-month-old boy presented with a painless tumor of the scalp, measuring about 2 cm in diameter. Case 2, a 3-year-old girl complained with a painless tumor of the knee, measuring approximately 1.5 cm in diameter. Microscopically, the tumor had a clear boundary and showed multinodular growth. The tumor was mainly composed of spindle cells arranged in long intersecting fascicles associated with thin, slit-like or branching ectatic vessels, focally forming hemangiopericytoma-like appearance. The tumor cells were abundant, but there was no obvious atypia. Mitotic figures (3-4/10 HPF) were noted. H-caldesmon and SMA were positive in both cases. Case 1 showed diffuse and strong positivity for Desmin, and focally for CKpan. Ki-67 proliferation index was 20% and 30%, respectively. FISH displayed NCOA2 gene translocation in case 1 and the RELA gene translocation in case 2. NGS detected the SRF-NCOA2 gene fusion in case 1 and the SRF-RELA gene fusion in case 2. Both patients underwent local excisions. During the follow-up of 5-14 months, case 1 had no local recurrence, while case 2 developed local recurrence 1 year post operatively. Conclusions: SRF-rearranged cellular perivascular myoid tumor is a novel variant of perivascular cell tumor, which tends to occur in children and adolescents. The tumor forms a broad morphologic spectrum ranging from a pericytic pattern to a myoid pattern, and include hybrid tumors with a mixture of pericytic and myoid patterns. Due to its diffuse hypercellularity and increased mitotic figures and smooth muscle-like immunophenotype, the tumor is easy to be misdiagnosed as myogenic sarcomas. The tumor usually pursues a benign clinical course and rare cases may locally recur.

[Efficacy of neoadjuvant therapy on HER2-positive breast cancer: a clinicopathological analysis].

Objective: To investigate the efficacy of neoadjuvant therapy (NAT) on HER2-positive breast cancer and to analyze their clinicopathological features. Methods: A total of 480 cases of HER2-positive breast cancer who received neoadjuvant therapy (NAT), diagnosed at the Department of Pathology of Fudan University Shanghai Cancer Center from 2015 to 2020, were retrospectively identified. Clinicopathological parameters such as age, tumor size, molecular subtype, type of targeted therapy, Ki-67 proliferation index, ER and HER2 immunohistochemical expression, and HER2 amplification status were analyzed to correlate with the efficacy of NAT. Results: Among 480 patients with HER2-positive breast cancer, 209 achieved pathology complete response (pCR) after NAT, with a pCR rate of 43.5%. Of all the cases,457 patients received chemotherapy plus trastuzumab and 23 patients received chemotherapy with trastuzumab and pertuzumab. A total of 198 cases (43.3%) achieved pCR in patients with chemotherapy plus trastuzumab, and 11 cases (47.8%) achieved pCR in patients with chemotherapy plus trastuzumab and pertuzumab. The pCR rate in the latter group was higher, but there was no statistical significance. The results showed that the pCR rate of IHC-HER2 3+patients (49%) was significantly higher than that of IHC-HER2 2+patients (26.1%, P<0.001). The higher the mean HER2 copy number in the FISH assay, the higher the pCR rate was achieved. The expression level of ER was inversely correlated with the efficacy of NAT, and the pCR rate in the ER-positive group (28.2%) was significantly lower than that in the ER-negative group (55.8%, P<0.001). The pCR rate (29.1%) of patients with luminal B type was lower than that of HER2 overexpression type (55.8%, P<0.001). In addition, higher Ki-67 proliferation index was associated with higher pCR rate (P<0.001). The pCR rate was the highest in the tumor ≤2 cm group (57.7%), while the pCR rate in the tumor >5 cm group was the lowest (31.1%). The difference between the groups was significant (P=0.005). Conclusions: HER2 copy numbers, HER2 immunohistochemical expression level, molecular subtype, ER expression level and Ki-67 proliferation index are significantly associated with pCR after NAT. In addition, fluorescence in situ hybridization results, HER2/CEP17 ratio and tumor size could also significantly affect the efficacy of NAT.

CLDN6 Suppresses Migration and Invasion of MCF-7 and SKBR-3 Breast Cancer Cells by Blocking the SMAD/Snail/MMP-2/9 Axis.

The study examined the mechanisms of action of signal protein claudin 6 (CLDN6) on migration and invasion of breast cancer cell lines MCF-7 and SKBR-3. To this end, the signal proteins SMAD were blocked with their inhibitor SB431542, the genes CLDN6 and SNAIL were knocked down with short hairpin RNAs, and MMP2 and MMP9 were inhibited with TIMP-1. Expressions of MMP2 and MMP9 mRNAs were evaluated by reverse transcription PCR, Expressions of MMP-2, MMP-9, E-cadherin, N-cadherin, and vimentin were examined by Western blotting. Migration and invasion were analyzed by scratch test and Matrigel invasion assay. SB431542 inhibited expression of MMP2 and MMP9 in both cell lines. Single use of SB431542 inhibited expression of MMP-2/MMP-9 and corresponding mRNAs, but subsequent silencing of CLDN6 gene reversed this effect. TIMP-1 reversed down-regulation of E-cadherin, upregulation of N-cadherin and vimentin, facilitation of migration and invasion evoked by CLDN6 knocking down. Silencing of SNAIL gene inhibited migration and invasion, upregulated the expression of E-cadherin, and down-regulated expression of MMP2, MMP 9, N-cadherin, and vimentin. Thus, CLDN6 suppresses the epithelial-mesenchymal transition, migration, and invasion via blocking SMAD/Snail/MMP-2/9 signaling pathway in MCF-7 and SKBR-3 cancer cell lines.

[EWSR1-SMAD3 positive fibroblastic tumor: a clinicopathological analysis].

Objective: To investigate the clinicopathological features, immunophenotypes and molecular genetics of EWSR1-SMAD3 positive fibroblastic tumor (ESFT) with an emphasis on differential diagnosis. Methods: The clinicopathological data, immunohistochemical profiles and molecular profiles of 3 ESFT cases diagnosed at the Department of Pathology, Fudan University Shanghai Cancer Center from 2018 to 2021were analyzed. The related literature was also reviewed. Results: There were two males and one female. The patients were 24, 12 and 36 years old, respectively. All three tumors occurred in the subcutis of the foot with the disease duration of 6 months to 2 years. The tumors were presented with a slowly growing mass or nodule, accompanied with pain in 1 patient. The tumors ranged in size from 0.1 to 1.6 cm (mean, 1.0 cm). Microscopically, the tumors were located in the subcutaneous tissue with a nodular or plexiform growth pattern. They were composed of cellular fascicles of bland spindle cells with elongated nuclei and fine chromatin. One of the tumors infiltrated into adjacent adipose tissue. There was no nuclear atypia or mitotic activities. All three tumors showed prominent stromal hyalinization with zonal pattern present in one case. Focal punctate calcification was noted in two cases. The immunohistochemical studies showed that tumor cells were diffusely positive for ERG and negative for CD31 and CD34, with Ki-67 index less than 2%. Fluorescence in situ hybridization on the two tested cases identified EWSR1 gene rearrangement. The next generation sequencing analysis demonstrated EWSR1-SMAD3 fusion in all three cases. During the follow up, one patient developed local recurrence 24 months after the surgery. Conclusions: ESFT is a benign fibroblastic neoplasm and has a predilection for the foot, characterized by ERG immunoreactivity and EWSR1-SMAD3 fusion. Local recurrence might occur when incompletely excised. Familiarity with its clinicopathological features is helpful in distinguishing it from other spindle cell neoplasms that tend to occur at acral sites.

[MAML2 gene rearrangement, fusion patterns and clinicopathological characteristics in primary pulmonary mucoepidermoid carcinoma].

Objective: To investigate MAML2 gene rearrangement, gene fusion patterns, and the clinicopathological characteristics of primary pulmonary mucoepidermoid carcinoma (PMEC). Methods: Forty-six cases of primary PMEC from Fudan University Zhongshan Hospital and Fudan University Shanghai Cancer Center between 2017 and 2020 were collected. MAML2 gene rearrangement in all cases was detected by fluorescence in situ hybridization (FISH). In 20 cases, MAML2 fusion patterns were detected by targeted RNA sequencing (RNAseq). The relationship between MAML2 gene rearrangement, fusion patterns, clinicopathological characteristics, and prognosis was analyzed. Results: The average age of PMEC patients was 41 years (range 15-71 years); the ratio of male to female was about 1.1 ∶ 1.0. Most PMECs were low grade in histopathology with an early clinical stage (stageⅠ-Ⅱ).The overall positive rate of MAML2 gene rearrangement detected by FISH was about 80.4% (37/46), and the rate was higher in low-grade PMEC (91.7%, 33/36). Of the 20 cases detected by RNAseq, all the 19 FISH positive cases showed gene fusion, mainly CRTC1-MAML2 fusion (16/19), the other three cases showed CRTC3-MAML2 fusion (3/19), the break point of all the fusion patterns was CRTC1/3 (exon 1)-MAML2 (exon 2); No gene fusion was detected in the single FISH negative case; Compared with the MAML2 FISH negative patients, the PMECs carrying CRTC1-MAML2 fusion were more commonly found in patients age ≤ 40 years, maximum tumor diameter ≤ 2 cm, low histopathological grade and early clinical stage (all P<0.05); The three PMECs carrying CRTC3-MAML2 fusion gene were all female with early clinical stage; Univariate analysis showed that MAML2 gene rearrangement/fusion, onset age ≤ 40 years old, smaller tumor size, low histopathological grade, early clinical stage, no metastasis at diagnosis and surgical treatment were significantly correlated with overall survival (P<0.05), but Cox regression analysis suggested that none of the above indicators were the independent prognostic factors for the survival of PMEC. Conclusions: The high incidence of MAML2 gene rearrangement in PMEC suggests that it is an important molecular diagnostic marker of PMEC. RNAseq confirms that CRTC1/3-MAML2 is the main fusion pattern in PMEC, suggesting that MAML2 fusion transcription may be an important driving factor of PMEC. MAML2 rearrangement/fusion and related clinicopathological characteristics are associated with good prognosis.

[Effect of imatinib on the height of children with chronic myeloid leukemia in the chronic phase].

Objective: To evaluate the effect of imatinib on growth impairment in children with chronic myeloid leukemia (CML-CP) in the chronic phase. Methods: From July 2018 to July 2019, questionnaires were distributed to CML children aged <18 years at the time of diagnosis who were receiving imatinib for at least 3 months or to their parents in China. The height-for-age standard deviation score (HtSDS) and the difference of standard deviation integral (△HtSDS) were used to explore the change in height with imatinib therapy. Results: The data of 238 respondents were included; 138 (58.0% ) respondents were men. The median age at the first diagnosis of CML was 11.0 years (range, 1.4-17.9 years) , and 93 (39.0% ) respondents were at the prepuberty stage. At the time of completing the questionnaires, the median age was 15.0 years (range, 2.0-34.0 years) . The median duration of imatinib therapy was 28 months (range, 3-213 months) . Among all the respondents, the mean HtSDS when completing the questionnaires (-0.063±1.361) was significantly lower than that at the time of starting imatinib treatment (0.391±1.244) (P<0.001) . Total 71.0% respondents showed growth impairment that was more common in those starting imatinib therapy at prepubertal age than in those starting at pubertal age. Multivariate analysis showed that younger at the start of imatinib therapy (P<0.001) and longer duration of imatinib therapy (P<0.001) were significantly associated with severe growth impairment on imatinib therapy. Conclusions: Imatinib induced growth impairment in children with CML-CP. Younger the age of initiation and longer the duration of imatinib therapy, more obvious the effect of imatinib on growth impairment.

[Ancillary values of fluorescence in situ hybridization with different gene combination in diagnosis of malignant melanoma].

Objective: To investigate the clinical value of the first multicolor fluorescence in situ hybridization (FISH) assay on multiple genes, and combined with 9p21 and 8q24 evaluation in the differential diagnosis of melanoma. Methods: Fifty-six melanomas and 36 benign melanocytic nevi diagnosed in Fudan University Shanghai Cancer Center from 2017 to 2019 were included. Each specimen was examined by first multicolor FISH assay targeting 6p25 (RREB1), 6q23 (MYB), 11q13 (CCND1) and CEP6, as well as 9p21 (CDKN2A) and 8q24 (MYC). The results of FISH assay in all cases were recorded according to Gerami's criteria. Basing on the sensitivity and specificity of the first FISH assay, the refinement of diagnosis by adding combined 9p21 and 8q24 probes was further evaluated, as well as their association with different clinicopathological features. Results: In 86 cases, the FISH signals were adequate for analysis. Of the 56 melanoma cases, 52 cases were adequate for analysis; 36 cases (69.2%) were positive in the first FISH assay. The most frequent chromosomal anomaly was gain of RREB1 (30/52, 57.7%), followed by gain of CCND1 (20/52, 38.5%), loss of MYB relative to CEP6 (18/52, 34.6%) and gain of RREB1 relative to CEP6 (17/52, 32.7%). The frequency of homozygous deletions in 9p21 was 15.4% (8/52) and gain of 8q24 was 36.5% (19/52). Among the 36 melanocytic nevi cases, FISH results could be accurately evaluated in 34 cases, and none showed a positive result in the first FISH assay or 9p21 and 8q24 FISH analysis. Compared with the first FISH assay, the sensitivity of combination with 9p21 and 8q24 FISH analysis increased from 69.2% to 76.9% (40/52) and the specificity remained 100.0%. Statistical data showed that the rates of FISH positivity in patients with acral-lentiginious melanoma and nodual melanoma subtypes were higher than that in patients with superficial spreading melanoma and lentigo maligna melanoma subtypes, and patients with Breslow thickness>2.0 mm had higher positive FISH frequency than patients with Breslow thickness ≤2.0 mm. Conclusion: Multisite FISH analysis is a highly effective ancillary tool for the differentiation of unequivocal malignant from benign melanocytic lesions. By combining the first FISH assay with CDKN2A and MYC assay, the clinical utility of FISH analysis is further optimized in differential diagnosis of melanoma. Patients with Breslow thickness>2.0 mm, or acral-lentiginious melanoma and nodual melanoma subtypes tend to have higher FISH positivity. There remains a need to further explore the ancillary value of FISH analysis in diagnosis of ambiguous lesions.

[Influenza virus activates toll-like receptor 7/nuclear factor-κB signaling pathway to regulate airway inflammatory response in patients with acute exacerbation of chronic obstructive pulmonary diseases].

Objective: To explore how influenza A virus (IAV) regulates airway inflammation via activating Toll-like receptor 7(TLR7)/nuclear factor of κB (NF-κB) signaling pathway in patients with acute exacerbation of chronic obstructive pulmonary disease (COPD). Methods: Primary bronchial epithelial cells were isolated and cultured from normal controls and COPD patients. Samples were divided into 6 groups according to different in vitro treatment, including normal epithelial cell group (A), normal cells+IAV group (B), COPD epithelial cell group (C), COPD cells+IAV group (D), normal cells+TLR7 small interference RNA (si-RNA) group (E), COPD cells+TLR7 siRNA group (F). Protein expressions of TLR7 and NF-κB were detected by Western blot after 24h co-culture with IAV and TLR7 siRNA. Interleukin-6 (IL-6) and tumor necrosis factor α (TNF α) were detected by enzyme-linked immunosorbent assay (ELISA). Results: (1) Compared with group A [0.350±0.075 and 0.470±0.034, (53.000±6.532)pg/ml and (17.000±1.625)pg/ml],TLR7, NF-κB protein expression and IL-6, TNF α levels were significantly increased in group B[0.950±0.075 and 1.090±0.078,(185.000±7.874)pg/ml and (32.000±0.838)pg/ml], group C[0.780±0.056 and 0.910±0.045,(138.000±5.100)pg/ml and 29.000±1.323)pg/ml) and group D[1.280±0.031 and 1.540±0.051,(432.000±5.734)pg/ml and (52.000±3.453)pg/ml] (all P<0.01). Compared with group C TLR7, NF-κB protein expression and IL-6, TNF α levels were significantly increased in group D (P<0.01). (2) Compared with the group A[0.530±0.023 and 0.800±0.046,(51.000±0.327)pg/ml and (14.000±0.314)pg/ml], TLR7, NF-κB protein expression and IL-6, TNF α levels were significantly decreased in the group E[0.350±0.047 and 0.510±0.067,(26.000±1.081)pg/ml and(8.000±0.526)pg/ml] (P<0.05). Compared with group C[1.080±0.078 and 1.280±0.034,(125.000±2.249)pg/ml and (28.000±1.010)pg/ml], TLR7, NF-κB protein expression and IL-6, TNF α levels decreased in the group F[0.880±0.056 and 1.040±0.029,(83.000±1.125)pg/ml and (21.000±0.429)pg/ml] (P<0.05). Conclusion: Influenza viruses activate TLR7/NF-κB signaling pathway to regulate airway inflammation storms in patients with acute exacerbation of COPD. New therapeutic targets of acute exacerbation COPD may be studied based on these inflammation responses to influenza viruses.

PM2.5 exposure induces alveolar epithelial cell apoptosis and causes emphysema through p53/Siva-1.

OBJECTIVE: This study aims to investigate whether PM2.5 exposure is involved in the induction of alveolar epithelial cell apoptosis and the progression of emphysema in mice, and to further explore its specific molecular mechanism. MATERIALS AND METHODS: A certain number of PM2.5 exposed mice and normal control mice were selected, and a lung resection operation was performed to collect the pulmonary tissue samples, which were then analyzed by hematoxylin and eosin (H&E) staining assay. Subsequently, the total protein in the pulmonary tissues of mice in PM2.5 exposure group and control group was extracted, and the p53 protein level was detected by Western blot. Meanwhile, in A549 cells, after treatment of different doses of PM2.5, the protein levels of p53, caspase3, and clv-caspase3 were examined by Western blot while the mRNA levels of p53, Siva-1, and clv-caspase3 were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR), respectively. In addition, flow cytometry was carried out to measure the incidence of cell apoptosis, while chromatin immunoprecipitation (ChIP) assay was performed to verify whether p53 binds to the Siva-1 promoter region and thus regulates its transcription process. RESULTS: H&E staining revealed that PM2.5 exposure caused pathological damage in the pulmonary tissues and the expansion of the spatial structure of alveoli, which led to emphysema in mice. Moreover, p53 protein expression in pulmonary tissue of mice in PM2.5 exposure group was remarkably higher than that in the control group. Subsequently, A549 cells were treated with 0, 25, 50, 100 µg/ml PM2.5 for 48 h, and it was found that, with the increase of PM2.5 exposure dose, the p53 protein level, Siva-1 mRNA level and cell apoptosis rate were all found increased in a dose-dependent manner, which could be partially reversed by transfection of si-p53 in A549 cells. In addition, CHIP experiments confirmed that p53 can bind to the Siva-1 promoter region and directly regulate Siva-1 transcription. In A549 cells, PM2.5 exposure increased the expression of the clv-caspase3 protein, which was reversed by the knockdown of p53; however, simultaneous overexpression of Siva-1 could further increase the clv-caspase3 protein level. Additionally, flow cytometry also revealed that PM2.5 exposure induced apoptosis of alveolar epithelial cells, while the knockdown of p53 reduced that, which could be promoted by the overexpression of Siva-1. CONCLUSIONS: PM2.5 exposure can promote the transcription of Siva-1 to induce apoptosis of alveolar epithelial cells and accelerate the progression of emphysema in mice by enhancing p53 protein expression.

[Clinicopathological study of BCOR rearrangement in high grade endometrial stromal sarcoma].

Objective: To investigate clinicopathological, cytogenetic features and differential diagnoses of high grade endometrial stromal sarcoma (HGESS) with BCOR gene rearrangement. Methods: Five cases of HGESS with BCOR rearrangement were collected from consultant files (2016-2018) at Fudan University Shanghai Cancer Center. Interphase FISH was performed using a dual color break-apart probe. The clinical data, histologic features and immunohistochemical findings were reviewed. Results: All 5 cases occurred in adult women with a median age of 48 (range, 45-55) years. Abdominal pain and abnormal vaginal bleeding were the most common symptoms. Microscopically, the tumors showed mainly tongue-like and/or intersecting myometrial invasion. Stromal myxoid matrix and/or collagen plaques were prominent in all the cases. Most tumors consisted of uniform, haphazard fascicles of short spindle cells with mild to moderate nuclear atypia. Mitotic figures and necrosis were easily identified. Significant nuclear pleomorphism was not seen. Most tumors were rich in thick-walled small vessels. Prominent perivascular tumor cell whorling seen in conventional low-grade endometrial stromal sarcoma was not seen. All tumors expressed CD10 with only focal or absent desmin, SMA and/or h-caldesmon staining. ER or PR expression was seen in 4 tumors and 1 tumor showed both marker expression. Diffuse cyclin D1 was present in 2 tumors. BCOR immunoreactivity was present with strong staining in 3 cases and moderate staining in 1 case respectively. Ki-67 index ranged from 10% to 30%. Fluorescence in situ hybridization confirmed chromosomal aberration of BCOR gene in all tumors, that were previously diagnosed as myxoid leiomyosarcoma (2 cases), spindle cell uterine sarcoma (2 cases) and low-grade endometrial stromal sarcoma (1 case). Limited follow-up information revealed that 3/5 patients developed tumor recurrence, metastasis or death within one year. Conclusion: BCOR rearranged HGESS has distinct morphological features and aggressive clinical behavior. In the presence of significant overlapping morphologic features between BCOR rearranged HGESS and other myxoid uterine mesenchymal tumors, especially myxoid leiomyosarcoma, molecular analysis is essential for accurate diagnoses.

Circulating microRNA array (miR-182, 200b and 205) for the early diagnosis and poor prognosis predictor of non-small cell lung cancer.

OBJECTIVE: Non-small cell lung cancer (NSCLC) is the most common cause for cancer-related mortality worldwide. Currently, early detection of NSCLC is one of the main available strategies for improving its prognosis. Due to the lack of non-invasive and convenient tools, early diagnosis of NSCLC remains poor. Recently, it has been reported that circulating microRNAs (miRNAs) can be stably detected in serum. Meanwhile, they play a powerful role as biomarkers in various tumors. Therefore, the aim of this study was to detect the expression levels of serum miR-182, 200b and 205 in NSCLC patients, and to investigate their diagnostic and prognostic values. PATIENTS AND METHODS: Real-time quantitative polymerase chain reaction (RT-qPCR) was carried out to measure the expressions of miR-182, 200b and 205 in NSCLC tissues and normal controls. Receiver-operating characteristic (ROC) curve analysis was performed to assess the potential value of serum miRNAs for NSCLC diagnosis. Meanwhile, transwell assays were performed to observe the functional effects of miRNAs on the invasion and migration of NSCLC cells. RESULTS: Compared with normal controls, serum levels of miR-182 and 205 in NSCLC patients were significantly upregulated, whereas miR-200b was remarkably downregulated. ROC analysis indicated that miRNA array (miR-182, 200b and 205) was useful biomarkers for early diagnosis of NSCLC. In addition, transwell assays demonstrated that miR-182 promoted the invasion and migration of NSCLC cells. CONCLUSIONS: Our findings revealed that serum miR-182, 200b and 205 might serve as promising biomarkers for early detection and treatment of NSCLC.

[Mammary myofibroblastoma: a clinicopathologic analysis of nine cases].

Objective: To study the clinicopathologic characteristics and differential diagnosis of mammary myofibroblastoma. Methods: Nine cases of mammary myofibroblastoma diagnosed between 2006 and 2017 were collected from the Department of Pathology, Fudan University Shanghai Cancer Center. Clinical and histopathologic features of these nine cases were examined, immunohistochemical staining was performed, FISH analysis for the detection of FOXO1 gene was performed in one case, and follow-up data were also collected. Results: There were seven female and two male patients, with a mean age of 54 years, median age of 50 years (ranging from 40 to 83 years). Four lesions each were located in the left and right breast, and one was in the left subaxillary accessory breast tissue. Clinically, 8 patients presented with a breast mass, 3 of which accompanied with pain. All of the tumors were well-demarcated grossly with a mean diameter of 2.5 cm. Microscopically, there were no entrapped ductal or lobular structures within the tumor. Seven tumors were classic type, which were composed of bland-looking spindle neoplastic cells without mitoses, arranging in intersecting fascicles, and interrupted by thick hyalinized collagen bundles. One case was of epithelioid variant, demonstrating epithelioid neoplastic cells diffusely arranged or in cluster. The other one case was mixed spindle and epithelioid-cell type. Atypical tumor cells were observed in 3 cases. Immunohistochemically, tumor cells were diffusely positive for desmin (9/9) and CD34 (6/9), as well as ER (7/7), PR (6/6) and bcl-2 (3/3). SMA (4/7) and Calponin (1/2) were focally or partially positive in some cases. H-caldesmon (1/2) was weakly positive and epithelial markers were negative. Ki-67 proliferation index was low (<10%). There was no monoallelic loss of FOXO1/13q14 loci in the detected case according to FISH analysis. Follow-up data were available for all patients, and follow-up period ranged from 12 to 78 months. All patients remained well without recurrence. Conclusions: Mammary myofibroblastoma is a rare benign mesenchymal tumor. In some circumstances, it may exhibit confusing morphologies, including some variants. The epithelioid variant of mammary myofibroblastoma might mimic invasive lobular carcinoma, leading to the diagnostic dilemmas and even misdiagnosis, especially in core needle biopsy specimen or frozen sections. Familiarity with the characteristics of this tumor is of great importance for accurate diagnosis and proper treatment.

[Angiofibroma of soft tissue: a clinicopathologic analysis of 24 cases].

Objective: To investigate the clinicopathologic and genetic features, pathologic diagnosis and differential diagnosis of angiofibroma of soft tissue(AFST). Methods: The clinicopathologic characteristics of 24 cases diagnosed at Fudan University Shanghai Cancer Center from 2011 to 2017 were analyzed; immunohistochemical staining and interphase fluorescence in situ hybridization (FISH) were performed, and the literatures were also reviewed. Results: There were 15 male and 9 female (male∶female=1.7∶1.0) patients with age of onset ranging from 8 to 68 years (mean, 45 years). Fourteen cases occurred in extremities, including upper limbs (n=3) and lower limbs (n=11); seven cases were in the trunk, and 1 case each was in the temporal region, retroperitoneum and liver, respectively. Clinically, the tumors usually presented as a slowly growing painless mass. Tumor sizes ranged from 0.8 to 14 cm (mean 4.6 cm). Microscopically, most lesions were well-circumscribed, with fibrous capsules. Few cases infiltrated the surrounding fibrofatty tissue focally. The tumors were mainly composed of sparse short spindle cells and numerous small, branching, thin-walled blood vessels distributed in amyxoid, fibromyxoid or collagenous matrix, often accompanied by medium-sized, round or irregular and ecstatic vessels at the tumor periphery.By immunohistochemistry, all tested cases expressed vimentin (5/5), and showed variable positivity for EMA (2/4), ER (1/2), PR (2/3), α-SMA (1/18)and desmin (1/10). Ki-67 proliferation index were all less than 5%. CD34, CD31 and ERG staining clearly outlined the contours of blood vessels in the stroma. Four cases were tested for NCOA2 gene rearrangement by FISH, of which three were positive. Follow-up data was available in 17 patients (range, 3 to 69 months; mean, 30 months) were all free of disease. Conclusions: Soft tissue angiofibroma is a benign fibroblastic neoplasm characterized by a prominent and complex vasculature set in a myxoid-to-collagenous stroma, and cytogenetically a distinctive NCOA2 gene rearrangement. Caution should be exercised for the possibility of potentially misinterpretation of AFST as vascular tumors and other myxoid soft tissue tumors.

Effects of propofol or sevoflurane anesthesia on the perioperative inflammatory response, pulmonary function and cognitive function in patients receiving lung cancer resection.

OBJECTIVE: To investigate the effects of propofol and sevoflurane anesthesia on the inflammatory response, pulmonary function and cognitive function of patients undergoing lung cancer resection and their differences. PATIENTS AND METHODS: 62 patients with lung cancer who underwent pulmonary lobectomy from January 2014 to January 2016 in Jining First People's Hospital were selected and randomly divided into two groups: the propofol group (n=31) and the sevoflurane group (n=31). Patients in the propofol group were treated with intravenous injection of propofol for anesthesia maintenance, whereas those in the sevoflurane group inhaled sevoflurane for anesthesia maintenance. All patients underwent surgical resection of the lobes by the same operator. Changes in the inflammatory response and pulmonary function of patients in the perioperative period were recorded before the induced anesthesia (t1), before one-lung ventilation (t2), after sternal closure by operation (t3) and at 24 h after operation (t4), respectively; the extubation time, eye opening time and response time of two groups of patients were recorded; mini-mental state examination (MMSE) was used to evaluate the changes in cognitive function in patients and detect the concentration of S100 calcium-binding protein ß (S100ß) in serum of patients before the induced anesthesia and at 24 h after operation, respectively. RESULTS: The difference of partial pressure of alveolar-arterial oxygen (A-aDO2), respiratory index (RI) and intra-pulmonary shunt fraction (Qs/Qt) of two groups of patients at t2 and t3 were significantly higher than those at t1 (p<0.01); during t2-t3, A-aDO2, RI and Qs/Qt of patients in the propofol group were significantly lower than those of patients in the sevoflurane group (p<0.05); the levels of interleukin-6 (IL-6) and matrix metalloproteinase-9 (MMP-9) in serum of patients after the induced anesthesia in the propofol group were significantly higher than those at t1, while the level of interleukin-10 (IL-10) was lower than that at t1 (p<0.01); during t2-t4, the levels of IL-6 and MMP-9 in serum of patients in the propofol group were significantly lower than those in patients in the sevoflurane group, while the level of IL-10 was significantly higher than that in patients in the sevoflurane group (p<0.05). The postoperative extubation time, eye opening time and response time of patients in the propofol group were significantly shorter than those of patients in the sevoflurane group (p<0.05). From intraoperative period to 24 h after operation, the prevalence rate of adverse reactions in patients in the propofol group was significantly lower than that in patients in the sevoflurane group (p<0.05); MMSE scores of two groups of patients at t4 were significantly lower than those at t1, while the concentration of S100ß was significantly higher than that at t1 (p<0.01); at t4, the MMSE score of patients in the propofol group was significantly higher than that in the sevoflurane group, while the concentration of S100ß was lower than that of patients in the sevoflurane group (p<0.05). CONCLUSIONS: Compared with sevoflurane anesthesia, propofol anesthesia can significantly reduce the perioperative inflammatory response in patients receiving lung cancer resection, shorten the recovery time after operation, protect the pulmonary function of patients, improve postoperative cognitive function, and reduce the prevalence rate of intraoperative adverse reactions.

MiR-616 promotes proliferation and inhibits apoptosis in glioma cells by suppressing expression of SOX7 via the Wnt signaling pathway.

OBJECTIVE: Accumulating evidence has indicated that miR-616 exerts tumor promoter roles in several types of cancer. However, the expression pattern and roles of miR-616 in glioma progression remain unknown. This study aimed to reveal the role of miR-616 in glioma cell proliferation and its potential mechanisms. PATIENTS AND METHODS: Real-time polymerase chain reaction was used to assay the expression of miR-616 in glioma tissue samples and glioma cell lines. MTT proliferation assay and flow cytometry analysis were performed to test the apoptosis and proliferation of glioma cell after down-regulation of miR-616. The target of miR-616 was predicted by TargetScan and confirmed by luciferase reporter assay. Changes in Wnt signaling markers expression were assessed using Western blotting. RESULTS: We found that the expression of miR-616 was increased in glioma tissues and cell lines. MTT and low cytometry analysis indicated that down-regulation of miR-616 significantly inhibited proliferation and promoted apoptosis in glioma cells. Moreover, SOX7 was confirmed to be a direct target of miR-616 in glioma cells using luciferase assay and Western blotting. Finally, it was found that down-regulation of miR-616 or upregulation of SOX7 could suppress the activity of Wnt/ß-catenin signaling in glioma cells. CONCLUSIONS: Our findings indicated that miR-616 acted as a tumor promoter in glioma, and its oncogenic roles were involved in the regulation of SOX7 and Wnt/ß-catenin signaling. Moreover, knockdown of miR-616 may provide a potential therapeutic strategy for glioma.

[Values of JAZF1 gene rearrangement detected by fluorescence in-situ hybridization in diagnosis of endometrial stromal tumours].

Objective: To investigate the role of JAZF1 gene rearrangement in the diagnosis and differential diagnosis of endometrial stromal sarcomas by fluorescence in situ hybridization (FISH). Methods: JAZF1 gene rearrangement was analyzed by FISH in 129 cases of ESS diagnosed from January 2008 to December 2016 including 105 cases of low-grade endometrial stromal sarcoma (LG-ESS), 21 cases of high-grade endometrial stromal sarcoma (HG-ESS) and 3 cases of undifferentiated uterine sarcoma (UUS). Sixteen cases of the related tumours in uterus were also collected as control group. The results were compared with our previous studies of JAZF1/JJAZ1 fusion gene in ESS by RT-PCR. Results: Detection of JAZF1 gene rearrangement by FISH was successfully analyzed in 144 cases. JAZF1 gene alteration was detected in 63 cases, all of which were LG-ESS, with an overall positivity of 60.6% (63/104), while no JAZF1 gene rearrangement was found in all other cases. JAZF1 gene rearrangement was present in LG-ESS with classic histology (69.3%, 52/75), smooth muscle differentiation (2/10), sex cord-like differentiation (4/5), fibromyxoid change (1/5), clear cell change (0/1), skeletal muscle differentiation (0/1), and schwannoma-like palisading pattern (0/1). The different components in all the cases of LG-ESS with variant histology had the clonal origin, with or without JAZF1 gene alteration. Compared to the results of JAZF1/JJAZ1 fusion gene by RT-PCR, the positive rate of JAZF1 gene rearrangement in LG-ESS by FISH (61.9%, 26/42) was significantly higher than that of RT-PCR (30.0%, 12/40; P<0.01). Conclusions: JAZF1 gene rearrangement is present only in LG-ESS, but not in HG-ESS, UUS or other related tumours in uterus. The frequency of JAZF1 gene rearrangement varies between classic LG-ESS and different morphologic variants. It is frequently, but not consistently, present in classic LG-ESS and less often positive in variant cases. The results of JAZF1 gene alterations in LG-ESS with different morphologic variants support the contention that the endometrial stromal and their variant morphologic components have the same clonal origin. Detection of JAZF1 gene rearrangement by FISH is very useful for the diagnosis and differential diagnosis of ESS.