Exogenous 6-Benzyladenine Improves Waterlogging Tolerance in Maize Seedlings by Mitigating Oxidative Stress and Upregulating the Ascorbate-Glutathione Cycle.

The synthetic cytokinin 6-benzyladenine (6-BA) regulates plant growth and prevents the negative consequences of various forms of abiotic stress, including waterlogging in crop plants. The present study aimed to investigate the effects of exogenous 6-BA on the growth, oxidative stress, and ascorbate-glutathione (AsA-GSH) cycle system in the inbred SY-MY13 (waterlogging-resistant) and SY-XT1 (waterlogging-sensitive) seedlings of waxy corn in conditions of waterlogging stress. The results demonstrated that waterlogging stress causes chlorosis and necrosis in waxy corn leaves, inhibiting growth and leading to the accumulation of reactive oxygen species (ROS), which induces oxidative stress and, in turn, reduces membrane lipid peroxidation and the disruption of membrane homeostasis. This is specifically manifested in the increased concentrations of superoxide anion radicals ( O 2 - ), hydrogen peroxide (H2O2), and malondialdehyde (MDA), in addition to increased relative electrical conductivity (REC%) values. The SY-MY13 strain exhibited growth superior to that of SY-XT1 when waterlogged due to its excellent waterlogging resistance. Thus, exogenous 6-BA was found to be effective in enhancing the growth of plants stressed by waterlogging in terms of the weight of the shoots and roots, shoot height, and leaf area. In addition to this, exogenous 6-BA also reduced the accumulation of O 2 - , H2O2, and MDA, increased ascorbate peroxidase (APX), glutathione reductase (GR), dehydroascorbate reductase (DHAR), and monodehydroascorbate reductase (MDHAR) activity, and enhanced ascorbic acid (AsA), and reduced glutathione (GSH) concentration through the regulation of the efficiency of the AsA-GSH cycle system in maize plants. Hence, the application of exogenous 6-BA can alleviate waterlogging-induced damage and improve waterlogging tolerance in waxy corn via the activation of the AsA-GSH cycle system and the elimination of ROS.

Diagnostic value comparison of CellDetect, fluorescent in situ hybridization (FISH), and cytology in urothelial carcinoma.

BACKGROUND: To evaluate the clinical effectiveness of a novel CellDetect staining technique, compared with fluorescent in situ hybridization (FISH), and urine cytology, in the diagnosis of urothelial carcinoma (UC). METHODS: A total of 264 patients with suspicious UC were enrolled in this study. All tissue specimens were collected by biopsy or surgery. Urine specimen was obtained for examinations prior to the surgical procedure. CellDetect staining was carried out with CellDetect kit, and FISH was performed with UroVysion detection kit, according to the manufacturer's instructions. For urine cytology, all specimens were centrifuged using the cytospin method, and the slides were stained by standard Papanicolaou stain. RESULTS: In this study, there were 128 cases of UC and 136 cases of non-UC, with no significant difference in gender and age between the two groups. Results for sensitivity of CellDetect, FISH, and urine cytology were 82.8%, 83.6%, and 39.8%, respectively. The specificity of the three techniques were 88.2%, 90.4%, and 86.0%, respectively. The sensitivity of CellDetect and FISH are significantly superior compared to the conventional urine cytology; however, there was no significant difference in specificity among three staining techniques. In addition, the sensitivity of CellDetect in lower urinary tract UC, upper urinary tract UC, non-muscle-invasive bladder cancer (NMIBC), and muscle-invasive bladder cancer (MIBC) were 83.3%, 81.8%, 83.5%, and 72.0%, respectively. The screening ability of CellDetect has no correlation with tumor location and the tumor stage. The sensitivity of CellDetect in low-grade UC and high-grade UC were 51.6 and 92.8%. Thus, screening ability of CellDetect in high-grade UC is significantly superior compared to that in low-grade UC. CONCLUSIONS: CellDetect and FISH show equal value in diagnosing UC, both are superior to conventional urine cytology. Compared to FISH, CellDetect is cost effective, easy to operate, with extensive clinical application value to monitor recurrence of UC, and to screen indetectable UC.

Comparative morphology of the internal elastic lamina of cerebral and peripheral arteries.

BACKGROUND: Atherosclerosis progresses later and with fewer complicated plaques in cerebral arteries than in peripheral arteries. The internal elastic lamina has been proposed to be important for the migration of smooth muscle cells into the intima during intimal thickening and atherosclerosis. METHODS: A total of 280 segments were retrieved from 14 autopsy specimens. Five sites were selected for analysis in each case: the middle cerebral artery, basilar artery, coronary artery, iliac artery and renal artery. We investigated the differences in the internal elastic lamina of cerebral and peripheral arteries. RESULTS: The average thickness of the internal elastic lamina of the cerebral arteries was larger than that of the peripheral arteries in both the early and advanced atherosclerotic plaque groups. Among the cerebral arteries, the basilar arteries had a thicker internal elastic lamina than the middle cerebral arteries. Among the peripheral arteries, the renal arteries had the thickest internal elastic lamina, followed by the iliac arteries and coronary arteries. Atherosclerosis led to a reduction in the thickness of the internal elastic lamina of the basilar, middle cerebral, and renal arteries. The stratification of the internal elastic lamina of iliac arteries significantly affected its measurement. The internal elastic lamina of coronary arteries was not affected by atherosclerosis, but it appeared fragmented. CONCLUSION: The results suggest that the characteristics of atherosclerotic plaques in cerebral and peripheral arteries may be related to the characteristics of the internal elastic lamina.

Clinicopathological analysis of epithelioid inflammatory myofibroblastic sarcoma.

Inflammatory myofibroblastic tumor (IMT) is a distinctive neoplasm composed of myofibroblastic and fibroblastic spindle cells, accompanied by the inflammatory infiltration of plasma cells, lymphocytes and/or eosinophils. Epithelioid inflammatory myofibroblastic sarcoma (EIMS), which primarily consists of cells with a round or epithelioid morphology, is associated with a poor prognosis and rapid development of local recurrence, and has been recognized to be a variant of IMT. Diagnosis of EIMS is difficult owing to its close resemblance to malignant mesothelioma, anaplastic large cell lymphoma, gastrointestinal stromal tumor and other malignant diseases. In the present study, a case of this rare tumor was evaluated in a 26-year-old male who was admitted to hospital after experiencing abdominal pain for ~18 days and abdominal distention for 1 week. The patient's tumor was examined by imaging, gross examination, histology, immunohistochemistry and fluorescence in situ hybridization (FISH). The magnetic resonance imaging enhanced-scanning image revealed that the morphology of the tumor was irregular, and signal was medley consisting of high and low hybrid reinforcement. Tumors were located in the bladder and rectal pit, in the lower part of the lower abdomen, indicating the presence of malignancy and involvement of the small intestine and rectum. Enhanced-scanning imaging revealed notable inhomogeneous enhancement. Gross examination revealed that the tumor was solid and had a variegated appearance with alternating fleshy and mucoid areas in the cut surface. Microscopically, the tumors were dominated by sheets of epithelioid-to-round cells with a prominent inflammatory infiltrate. The majority of the stroma was myxoid. Immunohistochemically, the tumor cells exhibited diffuse strong staining for ALK receptor tyrosine kinase (hereafter ALK), vimentin, tumor protein P53, desmin, Wilms' tumor 1 and programmed death-ligand 1. FISH analysis also revealed the existence of ALK rearrangement. The expression of PD-L1 in EIMS indicates that the immune checkpoint blockade could represent a novel therapy for the treatment of EIMS.

Expression and proliferation-promoting role of lymphoid enhancer-binding factor 1 in human clear cell renal carcinoma.

Lymphoid enhancer-binding factor 1 (LEF1) has been regarded as an important gene for carcinogenesis in many malignancies, however, the role of LEF1 in the progression of human renal cell carcinoma (RCC) has not been well studied. In this study, we investigated the expression of LEF1 in human RCC and the effect on proliferative ability of RCC cells. RCC samples from 138 patients who underwent radical nephrectomy were used in this study, the expression of LEF1 protein was determined by immunohistochemistry and Western blot, mRNA expression was analyzed by RT-PCR and real-time PCR. To investigate the effect of LEF1 on the proliferation of RCC cells, a LEF1 vector was transfected into RCC cells and LEF1 expression was also decreased by using siRNA. Proliferative ability of RCC cells was examined by WST-1 assay and a xenograft study with BALB/C nude mice. Our results indicated that LEF1 expression was significantly increased in stage III, IV and grade 3 RCC than in normal kidney, however, decreased LEF1 expression was found in low-stage and grade RCC compared to that in normal kidney, the expression of LEF1 was correlated to tumor stages, histologic grade, and tumor sizes in RCC. The effect of LEF1 on the proliferation in RCC was also analyzed, our results suggested that RCC cells expressing high levels of LEF1 had significantly increased proliferative ability compared to control cell lines, in contrast, RCC cells with a low LEF1 expression had lower proliferative ability. Moreover, LEF1 promoted proliferation of RCC cells depending on suppressing G2/M cell-cycle arrest. Our study demonstrated that the expression of LEF1 is associated with the progression of RCC and that LEF1 maybe involved in the development of RCC, these suggested LEF1 play a key role and might serve as a therapeutic target in treating advanced RCC.

[Molecular genetic abnormalities of N-myc and C-myc in pediatric neuroblastic tumors and clinical pathologic significance].

OBJECTIVE: To investigate the molecular genetic abnormalities of N-myc and C-myc, and their clinical pathological implications in pediatric neuroblastic tumors (NTs). METHODS: Abnormalities of N-myc were detected by interphase fluorescence in situ hybridization (FISH) technique in 246 cases of NTs, including neuroblastoma (NB,188 cases), ganglioneuroblastoma (GNB, 52 cases), ganglioneuroma (GN, 6 cases), and their association with the histological typing of the tumors and prognosis was analyzed. Abnormalities of C-myc were detected by FISH in 133 cases of NTs. RESULTS: Of the 246 cases of NTs, N-myc amplification was only found in 27 cases (11.0%, 27/246) of NB, but not in any cases of GNB or GN (P < 0.05). 89.0% (219/246) N-myc non-amplification were found in NTs, and it included N-myc gain in 175 cases (71.1%, 175/246) and normal N-myc in 44 cases (17.9%, 44/246). Univariate analysis indicated significantly (P = 0.012) poorer outcome in patients with N-myc amplification than N-myc non-amplification. However no significant difference was observed between N-myc gain cases and normal N-myc cases (P = 0.057). C-myc gain was found in 74 of 133 cases (55.6%) of NTs; no C-myc amplification or translocation was detected. Forty percent (6/15) of cases with N-myc amplification and 57.6% (68/118) of cases with N-myc non-amplification were accompanied by C-myc gain. The difference between N-myc amplification and non-amplification with C-myc gain was not significant (P > 0.05). Univariate analysis indicated that the outcome difference was not statistically significant between C-myc gain cases and normal C-myc cases (P = 0.357). CONCLUSIONS: The incidence of N-myc amplification only found in NB is low in pediatric NTs in China. Patients with N-myc amplification predict poorer outcome. No amplification or translocation of C-myc is detected in NTs, whereas C-myc gain is relatively common in NTs. There is no obvious association between N-myc amplification and C-myc gain.

ALK amplification and protein expression predict inferior prognosis in neuroblastomas.

BACKGROUND: ALK gene has been identified as a major neuroblastoma (NBL) predisposition gene. But ALK gene copy number and protein expression in ganglioneuroblastoma (GNBL) and ganglioneuroma (GN) are poorly described in the literature. Furthermore, there are controversies on the correlation between ALK protein expression and clinical outcome in NBL. METHODS: We evaluated MYCN/ALK gene copy number by fluorescence in situ hybridization (FISH) and detected ALK protein expression by immunohistochemistry (IHC) in 188 NBL, 52 GNBL and 6 GN samples and analyzed their association with clinical outcome of the patients. RESULTS: Although ALK gene copy number increase is a recurrent genetic aberration of neuroblastic tumors (NTs) (39.1%, 96/246), ALK amplification was only present in three NBLs (1.2%, 3/246). The frequency of ALK positivity in NBL (50.5%, 51/101) was significantly higher than in GNBL (22.6%, 7/31) and in GN (0.0%, 0/4) (P<0.05). In addition, ALK positivity also significantly correlates with MYCN/ALK gene copy number increases (P<0.05). Kaplan-Meier survival analysis indicated that MYCN/ALK amplification is correlated with decreased overall survival in NBL. A better prognosis trend was observed in patients with MYCN/ALK gain tumors compared with those with MYCN/ALK normal tumors. Furthermore, ALK positivity significantly correlated with inferior survival in NBL (P=0.044). CONCLUSION: ALK positivity in NTs correlated with advanced tumor types and MYCN/ALK gene copy number increases. ALK positivity predicts inferior prognosis in NBL and IHC is a simplified strategy to screen ALK positivity in clinical practice.

Protein tyrosine phosphatase ζ enhances proliferation by increasing ß-catenin nuclear expression in VHL-inactive human renal cell carcinoma cells.

OBJECTIVE: We investigated the role of protein tyrosine phosphatase ζ (Ptprz1) in human renal cell carcinoma (RCC) cells' proliferation and associations between Ptprz1 expression and von Hippel-Lindau (VHL) activation. METHODS: A normal human renal cell line and four human RCC cell lines were used in this study. VHL or Ptprz1 expression in RCC cells was increased by transfection with a VHL or Ptprz1 vector. VHL or Ptprz1 expression was decreased in these cells by siRNA using Lipofectamine 2000. Cells' proliferative activity was assessed by WST-1 assay. RESULTS: Our results suggested that Ptprz1 was a target of VHL, and a loss of VHL activation increased Ptprz1 expression in RCC cells. Ptprz1 enhanced ß-catenin protein expressions in the nuclear fractions of RCC cells and participated in regulating proliferation by activating ß-catenin and its downstream genes. In addition, a loss of VHL activity may enhance the proliferative activity of RCC cells by increasing Ptprz1 expression. CONCLUSION: Ptprz1-enhanced RCC cells' proliferation depends on VHL inactivation, and the Ptprz1/ß-catenin pathway may be a potential target for treating RCC with inactive VHL.

The influence of anthracosis and p16 ink4a gene aberrant methylation on small-sized pulmonary adenocarcinoma.

AIMS: Anthracosis is the deposition of black dusty material in the pulmonary parenchyma. Previous reports showed anthracosis and p16(ink4a) gene aberrant methylation are closely related to the promotion and progression of small-sized pulmonary adenocarcinoma. In this study, we investigated the influence of anthracosis and p16(ink4a) gene aberrant methylation on clinical samples from patients with small-sized adenocarcinoma. METHODS AND RESULTS: DNA was bisulfite modified and methylation-specific PCR was performed to detect p16(ink4a) gene aberrant methylation; black dusty material was extracted from lung tissues. Anthracotic index (AI) was defined as the absolute absorbance by densitometry. The histopathological diagnosis was concluded according to Noguchi's classification for small-sized pulmonary adenocarcinoma. The mean AI and the frequency of p16(ink4a) gene aberrant methylation of heavy smokers were significantly higher than that of nonsmokers (ï¼°<0.01 andï¼°<0.05, respectively). The frequency of p16(ink4a) gene aberrant methylation of early stage small-sized adenocarcinoma was lower than that of advanced and poorly differentiated, while p16(ink4a) protein expression level of early stage small-sized adenocarcinoma was significantly higher than that of poorly differentiated small-sized adenocarcinoma (P<0.05). CONCLUSIONS: AI and p16(ink4a) gene aberrant methylation may provide a potential universal biomarker for small-sized adenocarcinoma.

[Diagnosis and treatment of atypical adenomatous pulmonary hyperplasia in lungs].

OBJECTIVE: To analyze the characteristic of atypical adenomatous hyperplasia (AAH) in lungs though its computerized tomography (CT) scan, pathology and surgical mode. METHODS: The investigators retrospectively evaluated 10 atypical adenomatous hyperplasias (AAH) that were histologically confirmed and that manifested pure ground glass opacity (GGO) on thin-section helical CT scans. There were 2 males and 8 females with a median age 54.4 years old. All patients had the surgery. Their characteristic of CT scan and pathology were compared. All received a follow-up. RESULTS: In all cases, peripheral nodules were located at left upper lobe (n = 4), left lower lobe (n = 2) and right lower lobe (n = 4). GGO at a diameter of 0.5 - 1.2 cm was manifested on thin-section helical CT scans. The borderline of GGO was distinct and there was an equal density. Two of 10 cases underwent a wedge resection and 8 lobectomy. Postoperative patients recovered quickly without severe complications. Microscopically it manifested an apparent local pattern of alveolus epithelium hyperplasia in lungs. The alveolar interval had a slight increase. Local hyperplasia of fibrous cells was present with a slight degree of nucleus heteromorphism. The follow-up period was 2 months to 5 years. Ten patients with an excellent life quality survive without recurrence and metastasis. CONCLUSION: The preoperative diagnostic rate of AAH is boosted by high-differentiation enhancement CT scan and CT number histograms. But a definite diagnosis still requires the histological evidence. Surgery is one of the most reliable therapy for AAH.

[The influence of anthracosis and p16(ink4a) gene abberant methylation to the oncogenesis and progression of small pulmonary adenocarcinoma.].

BACKGROUND: Anthracosis is black dust matter deposition in the pulmonary parenchyma, which can cause bronchial deformity and destruction. Previously reported, anthracosis is closely correlated to the oncogenesis and progression of small pulmonary adenocarcinoma and p16(ink4a) gene aberrant methylation was closely associated with lung carcinogenesis. In this study, we want to characterize the influence of anthracosis and p16(ink4a) gene aberrant methylation on small adenocarcinoma. METHODS: DNA was bisulfite modified and then Methylation Specific PCR was used to detect p16(ink4a) gene aberrant methylation, and black dust matter was extracted from lung tissues, the absolute absorbance (A) detected by densitometry was defined as anthracotic index (AI). The histopathologic diagnosis was according to Noguchi's classification for small pulmonary adenocarcinoma. RESULTS: For heavy smokers, the mean AI was significantly higher than that of nonsmokers (P =0.005) and the frequency of p16(ink4a) gene aberrant methylation was also significantly higher than that of nonsmokers (P =0.023). The frequency of p16(ink4a) gene aberrant methylation of early stage small adenocarcinoma was lower than that of advanced and poor differentiated small adenocarcinoma, otherwise p16(ink4a) protein expression of early stage small adenocarcinoma was significantly higher than that of poor differentiated small adenocarcinoma (P =0.032). CONCLUSIONS: AI and p16(ink4a) gene aberrant methylation detection could be used as a combined potential biomarker of small adenocarcinoma.

Loss of function of p16 gene and prognosis of pulmonary adenocarcinoma.

BACKGROUND: Stepwise progression of peripheral-type lung adenocarcinoma was characterized morphologically and was related to prognosis. Expression of the tumor suppressor gene p16 in pulmonary adenocarcinoma decreased, mainly as a result of aberrant methylation of the CpG islands of the promoter region. METHODS: Aberrant methylation status of the p16 promoter region, the expression of its product, and loss of heterozygosity (LOH) on 9p21 were examined in surgically resected lung specimens from 57 patients (28 males and 29 females) with peripheral-type lung adenocarcinoma measuring

Anthracotic index and DNA methylation status of sputum contents can be used for identifying the population at risk of lung carcinoma.

BACKGROUND: Sputum cytology for the mass screening of lung carcinoma is a noninvasive, repeatable, and useful examination, but the detection rate is usually < 0.05% and the reliability is not high. METHODS: The anthracotic index (AI) and methylation status of the promoter regions of the p16, adenomatous polyposis coli (APC), and retinoic acid receptor-beta (RARbeta) genes were examined in 356 sputum specimens after routine cytologic examination. RESULTS: The mean AI of specimens from males was significantly higher than that from females. AI increased with increasing age and smoking index. The mean AI of patients with lung carcinoma was significantly higher than that of the nonaffected population. Furthermore, the mean AI of the specimens with or without cancer cells from patients with cancer was significantly higher than that of the nonaffected population. Abnormal methylation of the p16, APC, and RARbeta genes was detected in 21.7%, 28.2%, and 26.9% of specimens from patients with cancer, respectively. These ratios were significantly higher than those of the nonaffected populations (0%, 3.9%, and 7.6%, respectively). The incidences of abnormal methylation of the three genes were not associated with histologic classification, smoking index, gender, age, or occupation. CONCLUSIONS: These findings suggested that the AI and abnormal methylation status were useful for identifying a population at risk of lung carcinoma using mass screening of cytology specimens.

The implication of background anthracosis in the development and progression of pulmonary adenocarcinoma.

In order to characterize the relationship between background anthracosis and pulmonary adenocarcinogenesis, surgically resected tissues of 66 cases of stage I pulmonary adenocarcinoma, 4 cm or less at their greatest dimension, were examined. These cases were diagnosed based on the classification of small-sized adenocarcinoma of the lung (Noguchi et al., Cancer 75, 1995). Thirteen cases were diagnosed as types A (localized bronchioloalveolar adenocarcinoma, LBAC) and B (LBAC with alveolar collapse), 40 cases as type C (LBAC with a focus of fibroblastic proliferation), 8 as type D (poorly differentiated adenocarcinoma) and 5 as types E (bronchial gland type adenocarcinoma) and F (true papillary adenocarcinoma). The 5-year survival rate of types A and B cases was 100%, while those of type C, type D and types E and F were 52%, 48% and 39%, respectively. Nuclear accumulation of abnormal p53 protein in non-replacement type adenocarcinomas (types D, E and F) was detected more frequently than that in replacement type adenocarcinomas (types A, B and C) (P < 0.05). In each case, black dusty material was extracted from tumorous lesions and non-tumorous regions and blotted onto a nitrocellulose membrane. The anthracotic index (AI) was calculated with a densitometer. AIs of non-tumorous regions in early and replacement type adenocarcinomas (types A and B) were significantly less than in relatively advanced (type C) and poorly differentiated (type D) adenocarcinomas (P < 0.05). These results indicated that adenocarcinoma developing in heavily anthracotic lungs readily progresses to an advanced stage, or that adenocarcinoma with a less favorable prognosis tends to develop in severely anthracotic lungs.