A nomogram integrating ferroptosis-and immune-related biomarker for prediction of prognosis and diagnosis in kidney renal clear cell carcinoma.

OBJECTIVE: Approximately 60% of patients with kidney renal clear cell carcinoma (KIRC) die within the first 2-3 years. The prognosis for patients with KIRC and its metastases is poor. Ferroptosis and providing immunity are novel treatment targets for several cancers, including KIRC. Therefore, it is important to identify suitable ferroptosis- and immune-related signatures to predict the prognosis and diagnosis of patients with KIRC. MATERIALS AND METHODS: The corresponding data of patients with KIRC were obtained from the Cancer Genome Atlas. Univariate and multivariate Cox regression analyses were used to screen candidate biomarkers in patients with KIRC. RESULTS: We found that four FI-DEGs (BID, MET, LTB4R, and HMOX1) were independently associated with the overall survival of patients with KIRC. The prognosis and diagnosis model constructed using these four biomarkers could predict the outcome of KIRC, as measured by the receiver operating characteristic analyses. CONCLUSIONS: We identified 4 FI-DEGs that could be used as biomarkers in patients with KIRC. The present study not only contributes to understanding the roles of ferroptosis and immunity in the development of KIRC, but also to the diagnosis and prognosis of KIRC, although it remains to be further studied.

Quasi-isentropic compression of LiH above 400 GPa using magnetocumulative generator.

The knowledge of high-pressure behavior of LiH is significant for the validation of fundamental theoretical models and applications in thermonuclear materials and potential energy supplies. The compressibility of 7LiH under isentropic compression at high pressure was investigated experimentally and theoretically. The experimental technique for quasi-isentropic compression with low-density materials was developed using the magnetocumulative generator CJ-100 and x-ray flash radiography. The x-ray images and extracted interface of the sample target in dynamic flash radiography experiments were obtained. According to each interface size of the target both before and after compression, the compression ratio of 7LiH and reference material aluminum was obtained. The density of the reference and using its known isentropic curve provide the pressure in the reference. The pressure in 7LiH was deduced from the pressure in the reference and using the calculated gradient correction factor. The quasi-isentropic data point at 438 GPa was obtained experimentally. A semiempirical three-term complete equation of state was constructed and validated for 7LiH using the theory of Mie-Grüneisen-Debye with experimental data from the literature. The quasi-isentrope data point is reasonably consistent with the theoretical results. The quasi-isentropic experimental techniques and results broaden the existing research scope and are practical and helpful to further validate theoretical models in the future.

Values of long noncoding RNA SAMMSON in the clinicopathologic features and the prognostic implications of human gastric cancer.

OBJECTIVE: The aim of this study was to explore the clinical significance of lncRNA-survival associated mitochondrial melanoma-specific oncogenic non-coding RNA (lncRNA-SAMMSON) in the development and clinicopathological parameters of gastric cancer (GC). PATIENTS AND METHODS: Tissue specimens were collected from GC patients who received treatment in our hospital. Real-time quantitative polymerase chain reaction (QRT-PCR) was used to determine lncRNA-SAMMSON expression. Small interfering RNA (siRNA) was transfected to suppress the expression of lncRNA-SAMMSON in vitro. Pearson's χ2-test  was used to investigate the interaction of lncRNA-SAMMSON with clinicopathological parameters of GC patients. Kaplan-Meier method and Log rank analysis were used to analyze the progression-free survival time and overall time of GC patients. Furthermore, transwell assay and wound healing assay were conducted to determine the invasion and migration abilities of GC cells, respectively. RESULTS: QRT-PCR results showed that lncRNA-SAMMSON was abnormally overexpressed in GC tissues and cells (p<0.05). Pearson's χ2-test illustrated that clinical stage, distant metastasis and lymph node metastasis were closely related to lncRNA-SAMMSON expression in GC patients (p<0.05). Kaplan-Meier survival analysis represented that GC patients with high lncRNA-SAMMSON expression had significantly shorter progression-free survival time and overall survival time (p<0.05). Transwell assay and wound healing assay proved that inhibition of lncRNA-SAMMSON in GC cells dramatically reduced the invasion and migration abilities of GC cells, respectively (p<0.05). CONCLUSIONS: LncRNA-SAMMSON played an important role in the development of GC, which might be regarded as a new target for the diagnosis and treatment of GC.

Quantitative proteome analysis reveals annexin A3 as a novel biomarker in lung adenocarcinoma.

Metastasis is a common phenomenon and the major lethal cause of lung adenocarcinoma (AdC). To discover novel potential biomarkers associated with lymph node metastasis and prognosis in lung AdC, we assessed differences in protein expression between primary lung AdC with (LNM AdC) and without lymph node metastasis (non-LNM AdC) using a quantitative proteomic approach. Laser capture microdissection was performed to purify the cancer cells from primary lung AdC tissues. The differential proteins between the pooled microdissected non-LNM AdC and LNM AdC tissues were identified by two-dimensional difference gel electrophoresis (2D-DIGE) coupled with mass spectrometry (MS). In this study, twenty proteins were found to be differentially expressed in two types of lung AdC. ANXA3, significantly up-regulated in LNM AdC compared with non-LNM AdC, was validated by western blotting. Immunohistochemistry showed that ANXA3 over-expression was frequently observed in LNM AdCs and matched lymph node metastases compared with non-LNM AdCs. ANXA3 over-expression was significantly associated with advanced clinical stage (p < 0.001) and lymph node metastasis (p < 0.001) and increased relapse rate (p < 0.001) and decreased overall survival (p < 0.001) in lung AdCs. Cox regression analysis indicated ANXA3 over-expression was an independent prognostic factor. Our results indicate that ANXA3 might serve as a novel biomarker for lymph node metastasis and prognosis in lung AdC, and play an important role in lung AdC progression.

Aging is associated with increased proliferation and decreased apoptosis in the colonic mucosa.

Although the incidence of colon cancer increases with advancing age, reasons for this increase are not fully understood. Earlier studies have demonstrated that in Fischer-344 rats, aging is associated with increased crypt cell production in the colon, an event considered to be central to the initiation of carcinogenesis. Apoptosis also plays a critical role in the development and progression of colon cancer. Therefore, we have examined the age-related changes in proliferation and apoptosis in the colonic mucosa of 4-5, 12-14, and 22-24 month-old Fischer-344 rats. We have observed that proliferative activity in the colon, as assessed by proliferating cell nuclear antigen immunoreactivity, is higher (50-80%) in 12-14 and 22-24 month-old rats than in their 4-6 month-old counterparts. In contrast, the number of apoptotic cells, (as determined by TdT-mediated dUTP nick-end labeling assay) in the colonic mucosa of 12-14 and 22-24 month-old rats are considerably lower (50-60%) than in 4-6 month-old animals. These changes are accompanied by a concomitant reduction (75%) in pro-apoptotic Bak and stimulation (200%) of anti-apoptotic Bcl-xL levels. Since activation of caspases is associated with initiation and maintenance of apoptosis, we also analyzed the levels of pro and active forms of caspase-3, 8 and 9. The levels of active forms of caspase-3, 8 and 9 are found to be considerably (60-80%) lower in the colonic mucosa of 22-24 month-old rats, compared to their younger counterparts. This is accompanied by decreased cleavage of poly(ADP-ribose) polymerase, a substrate for caspases. In conclusion, our data show that aging enhances proliferation, but attenuates apoptosis in the colonic mucosa. These changes may partly be responsible for the age-related rise in colorectal cancer.

Increased in vitro activation of EGFR by membrane-bound TGF-alpha from gastric and colonic mucosa of aged rats.

Although aging is associated with increased epidermal growth factor receptor (EGFR) tyrosine kinase activity in Fischer 344 rat gastric and colonic mucosa, the regulatory mechanisms for the age-related rise in EGFR tyrosine kinase are poorly understood. Transmembrane transforming growth factor-alpha (TGF-alpha) may modulate EGFR function through an autocrine/juxtacrine mechanism. The present study aimed to determine the contribution of membrane-bound precursors of TGF-alpha in enhancing EGFR activation in the gastric and colonic mucosa during aging. The extent of EGFR tyrosine phosphorylation, a measure of EGFR activation, was substantially higher (300--350%) in the gastric and colonic mucosa of 23- (aged) vs. 4-mo-old (young) Fischer 344 rats. This was accompanied by an increase (200--1,000%) in the relative concentration of 18- to 20-kDa membrane-bound precursor forms of TGF-alpha. The amount of TGF-alpha bound to EGFR was also higher (150-250%) in the gastric and colonic mucosa of aged vs. young rats. In vitro studies revealed that exposure of HCT 116 cells (a colon cancer cell line) to TGF-alpha from gastric and colonic mucosal membranes of aged rats caused a 200--250% higher activation of EGFR and extracellular signal-related kinases (p42/44) compared with young rats. Our data suggest that the membrane-bound precursor form(s) of TGF-alpha may partly be responsible for enhancing EGFR activation in the gastric and colonic mucosa of aged rats, probably though an autocrine/juxtacrine mechanism(s).

[Determination of 17-hydroxycorticosteroid by capillary zone electrophoresis].

OBJECTIVE: To detect and dissociate 17-hydroxycorticosteroids(17-OHCS) in urine by using high performance capillary zone electrophoresis(CZE) so as to solve the problem that phenolic compounds interfere with the determination of urine 17-OHCS by chromatographic-spectrophotometry. METHODS: Pigments were absorbed from samples by activated kaolin in which the internal standard was theophyline. Urine 17-OHCS was absorbed by neutral styrene-divinylbenzene resin. Thus pigments and phenolic compounds were separated roughly. Then 17-OHCS was washed down by alcohol and separated and detected by capillary zone electrophoresis. RESULTS: Urinary 17-OHCS was separated successfully. Urinary 17-OHCS had a good linearity in the range of 2-50 mg.L-1 (r = 0.994). The average recovery rate was 96.17%. The relative standard deviation was less than 5%. The consistence detection limit was 0.3 mg.L-1. CONCLUSIONS: Compared with chromatographic-spectrophotometry, the high performance capillary electrophoresis has much less interference and is more simple, rapid, and accurate.

Aging enhances G(1) phase in the colonic mucosa of rats.

Although in Fischer-344 rats, aging has been shown to be associated with increased crypt cell production in the colonic mucosa, no information is available about the responsible intracellular mechanisms for the age-related rise in colonic mucosal cell proliferation. To determine whether cell cycling events are affected by aging, the present investigation examines the age-related changes in Cdk2 activity and the regulation of this process in the colonic mucosa. Colonic mucosae from 4-, 13- and 24-month-old Fischer-344 rats were assayed for Cdk2 activity and protein expression of Cdk2, cyclin D1 and E, as well as p21(Waf1/Cip1) (total and the fraction bound to Cdk2), p53 and phosphorylated Rb. Kinase activity and protein levels of Cdk2, as well as cyclin D1 concentration in the colonic mucosa, rose steadily with advancing age. However, the levels of cyclin E in the colonic mucosa were found to be higher in 24-month-old than 13-month-old rats, compared to their 4-month-old counterparts. On the other hand, levels of mucosal p21(Waf1/Cip1) (total and the fraction bound to Cdk2), one of the universal inhibitors of Cdks, were found to be lower in aged than in young rats. This was accompanied by a parallel decrease in mucosal p53, a tumor suppressor protein that is known to regulate p21(Waf1/Cip1). Additionally, we observed that the levels of phosphorylated Rb protein, a form which is involved in regulating progression of cells through the S phase, are increased in the colonic mucosa of 24-month-old rats, but not in 13-month-old animals, when compared with their 4-month-old counterparts. Our data suggest that, G(1) to S phase transition, as well as progression through the S phase of the cell cycle are accelerated in the colonic mucosa of aged rats.

Induction of transcriptional activity of AP-1 and NF-kappaB in the gastric mucosa during aging.

Although aging enhances expression and tyrosine kinase activity of epidermal growth factor receptor (EGFR) in the gastric mucosa, there is no information about EGFR signaling cascades. We examined the age-related changes in mitogen-activated protein kinases (MAPKs) [extracellular signal-related kinases (ERKs), c-Jun NH(2)-terminal kinases (JNKs), and p38], an EGFR-induced signaling cascade, and activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) transcriptional activity in the gastric mucosa of 4- to 6-, 12- to 14-, and 22- to 24-mo-old Fischer 344 rats. AP-1 and NF-kappaB transcriptional activity in the gastric mucosa rose steadily with advancing age. This can be further induced by transforming growth factor-alpha. The age-related activation of AP-1 and NF-kappaB in the gastric mucosa was associated with increased levels of c-Jun, c-Fos, and p52, but not p50 or p65. Total and phosphorylated IkappaBalpha levels in the gastric mucosa were unaffected by aging. Aging was also associated with marked activation of ERKs (p42/p44) and JNK1. In contrast, aging decreased p38 MAPK activity in the gastric mucosa. Our observation of increased activation of ERKs and JNK1 in the gastric mucosa of aged rats suggests a role for these MAPKs in regulating AP-1 and NF-kappaB transcriptional activity. These events may be responsible for the age-related rise in gastric mucosal proliferative activity.

Requirement of both tyrosine residues 1330 and 1337 in the C-terminal tail of the RON receptor tyrosine kinase for epithelial cell scattering and migration.

RON is a receptor tyrosine kinase that mediates cell scattering, migration, and tubular formation. This study focused on the function of two tyrosines, Y1330 and Y1337, in the C-terminus of RON in regulating epithelial cell scattering and migration. Substitution of both tyrosine residues with phenylalanine causes complete loss of cell scattering and migration in kidney 293 cells. In contrast, single mutation of either tyrosine residue has no effect. We found that mutation at Y1330 or Y1337 alone does not significantly affect the association of RON with PI-3 kinase, whereas a double mutation abolishes the recruitment of substrates. RON-mediated cell migration was inhibited by PI-3 kinase inhibitor wortmannin. This effect was also achieved by a dominant inhibitory p85 of PI-3 kinase. We conclude that Y1330 and Y1337 are required for RON-mediated cell motility. By associating with PI-3 kinase, the Y1330-Y1337 docking site plays a critical role in transducing motile signals of RON.

Induction of G(1) checkpoint in the gastric mucosa of aged rats.

Although in Fischer 344 rats aging is found to be associated with increased gastric mucosal proliferative activity, little is known about specific changes in the regulatory mechanisms of this process. To determine whether changes in cell cycling events could partly contribute to the age-related rise in gastric mucosal proliferative activity, the present investigation examines changes in cyclin-dependent kinase (Cdk2) activity and the regulation of this process in the gastric mucosa of Fischer 344 rats aged 4 (young), 13 (middle aged), and 24 (old) mo. We observed that aging is associated with a progressive rise in activity and protein levels of Cdk2 in the gastric mucosa. This is also found to be accompanied by a concomitant increase in cyclin E but not cyclin D1 levels. On the other hand, the levels of p21(Waf1/Cip1) (total as well as the fraction associated with Cdk2), a nuclear protein that is known to inhibit different cyclin-Cdk complexes, are found to decline in the gastric mucosa with advancing age. In contrast, with aging, there was a steady rise in p53 levels in the gastric mucosa. We have also observed that the levels of phosphorylated retinoblastoma protein, a form that participates in regulating progression through the S phase, are markedly elevated in the gastric mucosa of aged rats. In conclusion, our data suggest that, in the gastric mucosa, aging enhances transition of G(1) to S phase as well as progression through the S phase of the cell cycle. However, the age-related decline in p21(Waf1/Cip1) in the gastric mucosa appears to be independent of p53 status.

[Long-term result in 61 patients with primary lung cancer treated by pneumonectomy].

Long-term result of 61 patients with primary lung cancer treated by pneumonectomy is reported. There were 55 males and 6 females. The ages ranged from 23 to 62 years with a mean of 47.7. Of these 61 patients, 49 died within ten years after pneumonectomy, 34 of them died of recurrence or metastasis and 8 of cardio-pulmonary diseases. The 5 and 10 year survival rates were 24.1% and 16%, significantly lower than those of 104 patients treated by lobectomy during the same period (42.6% and 33.3%), P less than 0.05). The 5 and 10 year survival rates of left pneumonectomy were statistically higher than those of right pneumonectomy (36.7% and 21.7% versus 8.3% and 9.5%, P less than 0.05). In this paper, the dissection of mediastinal lymph nodes and effect on postoperative cardiopulmonary functions are discussed. The postoperative radiotherapy, to the mediastinum, is advocated because thorough dissection of the mediastinal lymph nodes is very difficult. Sleeve-lobectomy instead of pneumonectomy is indicated for those patients with the tumor located in one lobe and the main bronchus involved.