[Clinicopathological features of SMARCA4-deficient lung adenocarcinoma: a study of 42 cases].

Objective: To investigate the clinicopathological characteristics and genetic mutations of SMARCA4-deficient lung adenocarcinoma. Methods: From January 2021 to April 2023 in the First Affiliated Hospital of Zhengzhou University, 42 cases of SMARCA4-deficienct lung adenocarcinoma were diagnosed and now analyzed. All cases were retrospectively studied using hematoxylin-eosin staining and immunohistochemistry. The clinicopathological features were reviewed. Next-generation sequencing (NGS) was performed to investigate the mutations of related genes. Results: Among the 42 cases, there were 35 biopsy and 7 surgical specimens. There were 38 males and 4 females. The male to female ratio was 9.5∶1.0, with an age range from 42 to 78 years. Thirty-three patients were smokers. Overall, 4 cases (9.5%), 2 cases (4.7%), 18 cases (42.9%) and 18 cases (42.9%) were at stages Ⅰ, Ⅱ, Ⅲ, and Ⅳ, respectively. Microscopically, all the cases were non-mucinous adenocarcinoma, without lepidic pattern. The morphology was diverse. Rhabdomyoid cells, tumor giant cells and tumor necrosis were present. Most of the tumor cells had eosinophilic cytoplasm and occasionally clear cytoplasm. Defined cell borders and variable cytoplasmic hyaline secretory globules could be found. Inflammatory cells infiltrated the tumor stroma. Immunohistochemistry showed 29 cases (69.0%, 29/42) expressed TTF1, 10 cases (40.0%, 10/25) expressed Napsin A, and 20 cases (100.0%, 20/20) expressed INI1. Forty cases (95.2%, 40/42) showed BRG1 loss in all tumor cells, while 2 cases (4.8%, 2/42) had partial BRG1 loss. PD-L1 (22C3) was positive in 59.2% of the cases (16/27). NGS revealed mutations in EGFR, ROS1, MET, RET and KRAS. Six cases (6/8) showed SMARCA4 mutation, while some cases were accompanied by mutations of TP53 (7/15), STK11 (4/8), and KEAP1 (1/8). Driver gene mutations were more common in women (P<0.05). Patients were followed up for 1-25 months. Four patients died and 20 patients' diseases progressed. Conclusions: SMARCA4-deficient lung adenocarcinoma lacks characteristic morphology. Most of them express TTF1 and harbor driver gene mutations. It is necessary to identify this subset of lung adenocarcinoma by carrying out BRG1 stain routinely on lung adenocarcinoma. These patients can then be identified and benefit from targeted therapies.

[Analysis of clinical features and etiological diagnostic indices of reproductive age women with hyperandrogenism].

Objective: To investigate the clinical features and the value of different diagnostic indices for etiology in reproductive age women with hyperandrogenism. Methods: The medical records of 96 reproductive age women with hyperandrogenism in the multi-disciplinary team of Peking University First Hospital from January 2020 to April 2021 were collected. The patients were divided into four groups based on final diagnosis: congenital adrenal hyperplasia (CAH) (n=8), polycystic ovary syndrome (PCOS) (n=67), idiopathic hyperandrogenism (n=13) and other specific diseases (n=8), respectively. The indices related to androgens in different groups were compared, and then their efficiency for diagnosis of CAH and PCOS were analyzed with receiver operator characteristic curve (ROC curve). Results: A total of 96 patients with hyperandrogenism were recruited, with the age of 19-45 (29±6) years old. Overall, 4.2% (4/96) of the patients were with single clinical hyperandrogenism, 56.3% (54/96) were with single laboratory hyperandrogenaemia and 39.6% (38/96) were with both. The breakdown into laboratory hyperandrogenaemia subtypes was as follows: only T elevation 22.8% (21/92), only A2 elevation 7.6% (7/92), none DHEAS elevation, only FAI elevation 5.4% (5/92) and elevation of more than one of the androgen indices mentioned above accounted for 64.1% (59/92). In the reasons of consultation, simple irregular menstruation (36.0%, 32/89) or accompanied by clinical hyperandrogenism with or without infertility (36.0%, 32/89) were the most common. As for primary visiting departments, Obstetrics and Gynecology accounted for 53.2% (51/96), and then Endocrinology as 39.5% (38/96). The 17-OHP level of CAH, PCOS and idiopathic hyperandrogenism group was 20.0 (8.2, 33.1), 1.1 (0.8, 1.4), 0.9 (0.8, 1.3) ng/ml, respectively. The androstenedione level in these groups was 6.3 (4.6, 8.7), 3.8 (2.9, 4.8) and 3.2 (2.7, 3.7) ng/ml, respectively. The 17-OHP and androstenedione levels of CAH group were significantly higher than that in PCOS or idiopathic hyperandrogenism group (all P<0.05). The ratio of LH and FSH in these three groups was 0.8(0.5, 1.0), 1.3(0.6, 1.9) and 0.6(0.3, 0.7), respectively. The ratio of LH and FSH was significantly higher in PCOS than that in idiopathic hyperandrogenism group (P=0.024), but yet there was no significant difference compared with CAH group (P>0.05). The AUC of ROC curve of 17-OHP for CAH diagnosis was 0.94, followed by androstenedione 0.83, whereas LH/FSH for PCOS diagnosis was only 0.63. Conclusions: Among the reasons of consultation in reproductive age women who visited our multi-disciplinary team for female hyperandrogenism, simple irregular menstruation or accompanied by clinical hyperandrogenism with or without infertility are the most common. PCOS accounts for the majority of different androgen excess disorders. 17-OHP is the most valuable parameter for the diagnosis of CAH and secondly androstenedione.

[Mediastinal T lymphoblastic lymphoma/leukemia: clinicopathological and prognostic analyses of 61 cases].

Objective: To investigate the clinicopathologic features and prognosis of mediastinal T lymphoblastic lymphoma/leukemia (T-LBL/ALL). Methods: Sixty-one patients with mediastinal T-LBL/ALL diagnosed at First Affiliated Hospital of Zhengzhou University from August 1, 2011 to December 31, 2018 were enrolled. Their clinical, pathological, imaging features and prognosis were retrospectively analyzed. Results: Of the 61 patients with mediastinal T-LBL/ALL, 46 were male and 15 were female, with a male to female ratio of approximately 3∶1, aged 5 to 71 years (median 24 years, average of 24.5 years). Radiological findings were mediastinal soft tissue masses (58 cases) or mediastinal multiple enlarged lymph nodes (1 case). The tumor had a diameter of 4.9 to 18.3 cm in size, and data of 2 cases was unavailable. The patient's main symptoms were superior vena cava syndrome (cough, dyspnea, facial or neck edema), shortness of breath and chest pain, while about 1/3 of patients developed B symptoms (high fever, night sweats or significant weight loss). All 61 cases were biopsy specimens, and 2 of the tumors were later resected. Histopathologic examination showed that the thymic tissue epithelial network structure was destroyed or completely disappeared. A large number of lymphocytoid tumor cells were diffusely infiltrative, with infiltration into adipose tissue, starry sky phenomenon, linear-like arrangement, interstitial collagen hyperplasia and tumor cell extrusion. Focal tumor necrosis was present in some cases. Tumor cells were overall small to medium in size. They had little cytoplasm, slightly distorted, round or oval-shaped nuclei, fine chromatin, and innocuous/small nucleoli. Immunohistochemical studies showed that the tumor cells expressed CD7 (100%, 33/33), TDT (93.4%, 57/61), CD99 (83.3%, 25/30), CD1a (4/7), CD10 (8/18), CD34 (13.2%, 5/38), but did not express B cell markers (CD20 and PAX5) or granulocyte monocyte marker (MPO). The Ki-67 proliferation index was usually greater than 50%. One case was tested for TCR clonal rearrangement, which was positive. Several hemotherapy regiments were used. Hyper-CVAD (cyclophosphamide, vindesine, dexamethasone, and epirubicin) were most frequently administrated (60.4%, 32/53), followed by BFM-90 (50.9%, 27/53). Some patients were treated with the above two and other treatment options. Follow-up data were available in 55 of the 61 patients, and 26 patients (47.3%) survived. The average five-year survival rate was 50.6%. The patient's prognosis was not significantly related to the International Prognostic Index, age of onset, gender, or tumor size. Conclusions: The mediastinal T-LBL/ALL is rare, and most of its specimens are needle biopsies. The histological morphology is often difficult to interpret, while the addition of clinical features and immunohistochemistry may help. The combination of CKpan, TDT, CD99, CD7, CD3, PAX5, CD34, CD10, and Ki-67 immunohistochemicl studies may assist in diagnosis of the most cases.

[Clinicopathological features of pulmonary epithelioid hemangioendothelioma: a study of 18 cases].

Objective: To investigate the clinicopathological features of pulmonary epithelioid hemangioendothelioma (PEHE). Methods: Eighteen cases of PEHE were collected from August 2011 to December 2018 at the First Affiliated Hospital of Zhengzhou University. All cases were retrospectively studied by hematoxylin and eosin staining and immunohistochemistry (IHC). The clinicopathological features were reviewed; the status of CAMTA1 and TFE3 gene was analyzed and patients' outcome was followed up. Results: Of the 18 cases, there were 11 males and 7 females with a male to female ratio of 1.6 to 1.0. The patients' age ranged from 36 to 68 years (mean 52 years). Twelve cases (12/18) showed a single nodule and six cases (6/18) showed multiple bilateral nodules. Seven cases (7/18) involved other organs besides lung. Seventeen (17/18) patients presented with respiratory symptoms and one patient (1/18) presented with abdominal pain. Grossly, the tumors were greyish-white nodules with indistinct borders. Microscopically the tumor cells were epithelioid and arranged in strands and nests, and cytoplasmic vacuoles were commonly noted. The stroma was myxochondroid or hyaline. By IHC, the tumor cells were positive for CD31(18/18), CD34 (16/18), ERG (18/18) and Fli-1 (18/18); CKpan was focally positive in 5 cases (5/18). TFE3 was positive in 3 cases (3/18), and Ki-67 index ranged from 5% to 30%. FISH analysis showed seventeen cases (17/18) had CAMAT1 rearrangement, one case had TFE3 rearrangement displaying a split signal. Eight patients (8/18) had surgical excision, three patients (3/18) had surgery and chemotherapy, and seven patients (7/18) had chemotherapy only. Four patients (4/18) died of the disease. Conclusions: Patients with PEHE have non-specific symptoms, and correct diagnosis depends on pathologic biopsy and the exclusion of other tumors with epithelioid morphology. Some patients with PEHE have poor prognosis, particularly in those who have multiple nodules, peripheral invasion or metastasis.

[MicroRNA-140-5p suppresses cell proliferation and invasion in esophageal squamous cell carcinoma by targeting Glut1].

Objective: To investigate the expression of microRNA-140-5p (miR-140-5p) in esophageal squamous cell carcinoma (ESCC) and its role in cell proliferation and invasion of ESCC. Methods: Real-time quantitative PCR (qPCR) was used to detect the expression levels of miR-140-5p in ESCC tissues and cells. Negative control and miR-140-5p mimic were transfected into Eca109 and KYSE70 cells. CCK-8 kit and Transwell assay were employed to examine the changes of cell proliferation and invasion ability after transfection, respectively. The dual-luciferase reporter assay was used to assess the interaction of miR-140-5p with Glut1. Western blot was utilized to detect the Glut1 protein expression after transfection. Results: Analysis of the related GEO datasets revealed that the expression of miR-140-5p in ESCC tissues was significantly lower than that in normal tissues (P<0.01). The qPCR testing demonstrated that the expression of miR-140-5p in ESCC tissues and cells was markedly lower than that in normal tissues and normal esophageal epithelial cell Het-1A (P<0.01). The miR-140-5p expression was closely associated with tumor differentiation, TNM staging and lymph node metastasis in ESCC patients. The survival rate of ESCC patients with high miR-140-5p level was higher than those with low miR-140-5p level (P<0.05). Besides, addition of miR-140-5p mimic significantly upregulated the expression of miR-140-5p in Eca109 and KYSE70 cells, and suppressed cell proliferation and invasion in Eca109 and KYSE70 cells. The dual-luciferase reporter assay showed that Glut1 was a direct target of miR-140-5p in ESCC cells, and its expression was upregulated in ESCC tissues. Glut1 expression was inversely associated with miR-140-5p expression in ESCC tissues. MiR-140-5p mimic dramatically inhibited the expression of Glut1 in Eca109 and KYSE70 cells. Conclusions: MiR-140-5p plays an essential role in ESCC development and progression. Targeting at miR-140-5p/Glut1 may be a novel therapeutic strategy for ESCC patients.

[Clinicopathological features and prognostic factors of primary pulmonary adenoid cystic carcinoma: a study of 59 cases].

Objective: To investigate the clinicopathological features and prognostic indicators of primary pulmonary adenoid cystic carcinoma. Methods: Fifty-nine cases of primary pulmonary adenoid cystic carcinoma were collected from August 2011 to December 2017 at the First Affiliated Hospital of Zhengzhou University. All cases were retrospectively studied by hematoxylin-eosin staining and immunohistochemistry. The clinicopathological features were reviewed and patient survival analysis was performed using Kaplan-Meier method and Cox regression model. Status of epidermal growth factor receptor (EGFR), KRAS, BRAF genes was analyzed in 15 of the 59 study cases. Results: Among 59 cases, there were 25 males and 34 females with male to female ratio of 1.0 to 1.4. The patient age ranged from 29 to 81 years with a mean age of 55 years. The tumor max diameters ranged from 1.0 to 9.6 cm with an average diameter of 2.8 cm. Fifteen (25.4%) patients were smokers while 44 patients (74.6%) were non-smokers. Tumors predominantly occurred in the trachea (28/59,47.5%), the left main bronchus (7/59,11.9%) and the right bronchus (5/59,8.5%). Grossly, the tumors were well circumscribed, greyish-white nodules. Microscopically the tumor cells were small and uniform, and arranged in tubular, cribriform, and solid patterns. Immunohistochemistry showed that the tumor cells were positive for CK7, S-100 protein, Sox-10, CD117 and p63. TTF1 was only positive in 2 cases and Ki-67 index ranged from 3% to 40%. Eighteen cases (30.5%) were gradeⅠ, 26 cases (40.1%) grade Ⅱ, and 15 cases (25.4%) grade Ⅲ. Overall, 39 cases (66.1%), 7 cases (11.9%), 10 cases (16.9%), and 3 cases (5.1%) were at stages Ⅰ, Ⅱ, Ⅲ, and Ⅳ, respectively. Twenty-three patients (39.0%) received surgical therapy, 3 patients (5.1%) surgery combined with radiotherapy, 9 patients (15.2%) surgery combined with chemotherapy, and 24 cases (40.7%) chemotherapy only. No mutation of EGFR, KRAS and BRAF was detected in all 15 tested cases. The overall survival rate at the first, third and fifth years was 94.9%, 86.4% and 84.7%, respectively. Prognostic analysis showed that patient's age and tumor size were statistically associated with the survival (P<0.05). Conclusions: Majority of the patients with primary pulmonary adenoid cystic carcinoma are at an early clinical stage with a favorable prognosis. The size of the tumor and the age of the patients are independent prognostic indicators.

[Tracheobronchopathia osteochondroplastica: a clinicopathological analysis of 18 cases].

Objective: To investigate the clinical symptoms, imaging features, pathologic manifestations and diagnosis of tracheobronchopathia osteochondroplastica (TO). Methods: The clinical data, imaging and pathologic features and outcome of 18 TO patients diagnosed at the First Affiliated Hospital of Zhengzhou University from August 2011 to August 2018 were collected and analyzed. Results: The 18 TO patients included 10 males and 8 females; patients' age range was 31 to 64 years (mean 52 years). Six patients (6/18) were smokers. The main presenting clinical symptoms included cough in 15 cases, expectoration in eight cases (8/18), hemoptysis in five cases (5/18), chest tightness in four cases, wheezing in three cases and chest pain in two cases. The time interval between the initial symptoms and diagnosis was 1.5 to 360.0 months, and the average time interval was 45.2 months. Blood calcium and phosphorus were normal in 18 patients (18/18). Chest X-ray showed no direct evidence of TO. Six patients (6/18) showed irregular changes in the trachea or bronchial wall by chest CT scan. Three patients (3/18) had mild ventilatory obstruction. TO was classified as: 10 cases (10/18) were scattered type, seven cases (7/18) were diffuse type and one case (1/18) was confluent type. Epithelial squamous metaplasia, submucosal cartilage, submucosal ossification and hematopoietic bone marrow within the ossified areas were the characteristic histopathologic findings of TO. Conclusions: TO is a rare benign disorder that shows atypical presentation. CT scan is insensitive, the histopathology shows submucosal cartilage or ossification. TO should be diagnosed by comprehensive consideration of clinical symptoms, imaging and pathology.

[Expression of BRD4 in squamous cell carcinoma and its effects on cell proliferation and invasion ability].

Objective: To investigate the expression of BRD4 in squamous cell carcinoma (SCC) tissues and cells, and the effects of its expression on cell proliferation and invasion ability. Methods: Immunohistochemistry was used to detect BRD4 protein expression in SCC tissues and paired normal esophageal squamous epithelial tissues. The expression of BRD4 protein was detected in different SCC cell lines and normal esophageal squamous epithelial cells by Western blot. BRD4 siRNA and control siRNA were used to transfect SCC Eca109 cells, and experiments were divided into three groups: untreated group, control siRNA group and BRD4 siRNA group. Western blot was employed to investigate the expression of BRD4 protein in the three groups of SCC Eca109 cells. CCK-8 kit was utilized to detect cell proliferation ability, and Transwell chamber was used to examine cell invasion ability. Finally, Western blot was used to detect the expression of MMP2 and MMP9 proteins. Results: The positive rate of BRD4 protein expression in SCC tissues was significantly higher than that of normal squamous epithelial tissues. The expression of BRD4 protein in 4 SCC cell lines was higher than that in normal esophageal cell Het-1A. BRD4 siRNA obviously downregulated the expression of BRD4 protein in Eca109 cells, and its downregulation contributed to the suppression of cell proliferation and invasion ability in Eca109 cells (all P<0.05), coupled with the decreases of MMP2 and MMP9 proteins. Conclusion: BRD4 may be closely associated with the proliferation and invasion of SCC, and it thus may be a potential therapeutic target for SCC.

Cloning and expression analysis of phytoene desaturase and ζ-carotene desaturase genes in Carica papaya.

The fruit flesh color of papaya is an important nutritional quality trait and is due to the accumulation of carotenoid. To elucidate the carotenoid biosynthesis pathway in Carica papaya, the phytoene desaturase (PDS) and the ζ-carotene desaturase (ZDS) genes were isolated from papaya (named CpPDS and CpZDS) using the rapid amplification of cDNA ends (RACE) approach, and their expression levels were investigated in red- and yellow-fleshed papaya varieties. CpPDS contains a 1749 bp open reading frame coding for 583 amino acids, while CpZDS contains a 1716 bp open reading frame coding for 572 amino acids. The deduced CpPDS and CpZDS proteins contain a conserved dinucleotide-binding site at the N-terminus and a carotenoid-binding domain at the C-terminus. Papaya genome sequence analysis revealed that CpPDS and CpZDS are single copy; the CpPDS was mapped to papaya chromosome LG6, and the CpZDS was mapped to chromosome LG3. Quantitative PCR showed that both CpPDS and CpZDS were expressed in all tissues examined with the highest expression in maturing fruits, and that the expression of CpPDS and CpZDS were higher in red-fleshed fruits than in yellow-fleshed fruits. These results indicated that the differential accumulation of carotenoids in red- and yellow-fleshed papaya varieties might be partly explained by the transcriptional level of CpPDS and CpZDS.

MiR-16 inhibits hepatocellular carcinoma progression by targeting FEAT through NF-κB signaling pathway.

OBJECTIVE: MicroRNA-16 (miR-16) expression has been proved to take part in the initiation and development of several cancers, including hepatocellular carcinoma (HCC). However, its role and its molecular mechanism in HCC cells remain unclear. Our study aimed to elucidate miR-16 probable role and potential mechanism in HCC cells. PATIENTS AND METHODS: MiR-16 expression in HCC was measured by Real Time-Polymerase Chain Reaction (RT-PCR). MiR-16 mimic or inhibitor was applied to increase or decrease miR-16 expression in Huh7 cells separately. The cell viability was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide). The invaded cells and migrated cells were detected by the transwell assay. The epithelial-mesenchymal transition (EMT) and the nuclear factor-κB (NF-κB) were performed using Western blot. The tumor growth was measured via xenograft tumor formation assay. Moreover, bioinformatical methods and luciferase reporter assay were carried out to confirm the miR-16 target gene. RESULTS: MiR-16 expression was downregulated in HCC tissues and cells. Furthermore, the increasing miR-16 expression was suppressed, whereas the decreasing miR-16 expression promoted cell proliferation, invasion, and migration in Huh7 cells. Moreover, miR-16 targeted FEAT in regulating HCC progression. FEAT was associated with a poor prognosis of HCC patients. Especially, miR-16 suppressed EMT and NF-κB pathway in HCC and inhibited the tumor growth in vivo. CONCLUSIONS: We stated that miR-16 suppressed HCC cell progression by targeting FEAT and inhibiting EMT and NF-κB pathway. MiR-16 may be clinically utilized as a factor for the clinical diagnosis and prognosis of HCC.

Harmony search method: theory and applications.

The Harmony Search (HS) method is an emerging metaheuristic optimization algorithm, which has been employed to cope with numerous challenging tasks during the past decade. In this paper, the essential theory and applications of the HS algorithm are first described and reviewed. Several typical variants of the original HS are next briefly explained. As an example of case study, a modified HS method inspired by the idea of Pareto-dominance-based ranking is also presented. It is further applied to handle a practical wind generator optimal design problem.

Sustained viability of acute myelogenous leukemia cells in liquid suspension culture correlates with occurrence of differentiation.

To examine the relationship between viability of acute myelogenous leukemia (AML) blasts in liquid suspension culture and the occurrence of myeloid maturation in culture, total and differential cell counts and viability as determined by trypan blue dye exclusion were followed over 21 days of culture. A significant positive correlation was found between viability of cells on day 21 of culture and percentage of mature cells present. These data suggest that myeloid maturation may be a mechanism by which AML cells maintain viability in an adverse environment.