GRP78 facilitates M2 macrophage polarization and tumour progression.

This study investigated the regulation of GRP78 in tumour-associated macrophage polarization in lung cancer. First, our results showed that GRP78 was upregulated in macrophages during M2 polarization and in a conditioned medium derived from lung cancer cells. Next, we found that knocking down GRP78 in macrophages promoted M1 differentiation and suppressed M2 polarization via the Janus kinase/signal transducer and activator of transcription signalling. Moreover, conditioned medium from GRP78- or insulin-like growth factor 1-knockdown macrophages attenuated the survival, proliferation, and migration of lung cancer cells, while conditioned medium from GRP78-overexpressing macrophages had the opposite effects. Additionally, GRP78 knockdown reduced both the secretion of insulin-like growth factor 1 and the phosphorylation of the insulin-like growth factor 1 receptor. Interestingly, insulin-like growth factor 1 neutralization downregulated GRP78 and suppressed GRP78 overexpression-induced M2 polarization. Mechanistically, insulin-like growth factor 1 treatment induced the translocation of GRP78 to the plasma membrane and promoted its association with the insulin-like growth factor 1 receptor. Finally, IGF-1 blockade and knockdown as well as GRP78 knockdown in macrophages inhibited M2 macrophage-induced survival, proliferation, and migration of lung cancer cells both in vitro and in vivo.

Investigation of Factors Influencing the Fear of Cancer Recurrence in Breast Cancer Patients Using Structural Equation Modeling: A Cross-Sectional Study.

Objective: This study aimed to investigate the fear of cancer recurrence (FCR) in breast cancer patients and develop a structural equation model of influencing factors to help formulate clinical intervention strategies. Methods: A convenience sample of 325 patients was surveyed using a general and disease-related data questionnaire, which combined the Fear of Progression Questionnaire-Short Form, Mishel Uncertainty in Illness Scale, Perceived Social Support Scale, and Medical Coping Modes Questionnaire. Results: The total score of FCR in breast cancer patients was 35.06 ± 10.83, and 53.8% of patients reached the clinical level. The structural equation model demonstrated that illness uncertainty had a direct positive impact on FCR (ß = 0.275, p < 0.05), and it could have an indirect impact through social support and resignation coping methods (ß = 0.254, p < 0.05). Conclusion: The fear of cancer recurrence in breast cancer patients needs further understanding. Medical staff can reduce or buffer FCR in breast cancer patients by strengthening positive influences, such as social support, or weakening negative influences, such as illness uncertainty and resignation coping.

Autophagy in pulmonary hypertension: Emerging roles and therapeutic implications.

Autophagy is an important mechanism for cellular self-digestion and basal homeostasis. This gene- and modulator-regulated pathway is conserved in cells. Recently, several studies have shown that autophagic dysfunction is associated with pulmonary hypertension (PH). However, the relationship between autophagy and PH remains controversial. In this review, we mainly introduce the effects of autophagy-related genes and some regulatory molecules on PH and the relationship between autophagy and PH under the conditions of hypoxia, monocrotaline injection, thromboembolic stress, oxidative stress, and other drugs and toxins. The effects of other autophagy-related drugs, such as chloroquine, 3-methyladenine, rapamycin, and other potential therapeutic drugs and targets, in PH are also described.

Charge transfer effect on Raman shifts of aromatic hydrocarbons with three phenyl rings from ab initio study.

To clarify the charge transfer effect on Raman spectra of aromatic hydrocarbons, we investigate the Raman shifts of phenanthrene, p-terphenyl, and anthracene and their negatively charged counterparts by using density functional theory. For the three molecules, upon charge increasing, the computed Raman peaks generally shift down with the exception of a few shifting up. The characteristic Raman modes in the 0-1000 cm-1 region persist up, while some high-frequency ones change dramatically with three charges transferred. The calculated Raman shifts for one- and two-electron transfer are in agreement with the measured Raman spectra, and in accordance to the stoichiometric ratios 1:1 and 2:1 of the metal atom and aromatic hydrocarbon molecule in recent experimental and theoretical studies. Our theoretical results provide the fundamental information to elucidate the Raman shifts and the stoichiometric ratios for alkali-metal-doped aromatic hydrocarbons.

MicroRNA-214-3p Regulates Hypoxia-Mediated Pulmonary Artery Smooth Muscle Cell Proliferation and Migration by Targeting ARHGEF12.

BACKGROUND miR-214-3p has been found to inhibit proliferation and migration in cancer cells. The objective of this study was to determine whether ARHGEF12 is involved in miR-214-3p-mediated suppression of proliferation and migration of pulmonary artery smooth muscle cells (PASMCs). MATERIAL AND METHODS PASMCs were cultured under normoxia or hypoxia. miR-214-3p mimics or inhibitors were transiently transfected into PASMCs. Proliferation, apoptosis, and migration of PASMCs were evaluated using MTT assay, flow cytometry, and Boyden chamber apparatus. Western blot analysis was used to examine expression of Rho guanine nucleotide exchange factor 12 (ARHGEF12), c-fos, c-jun, and caspase-3. Luciferase reporter assay was used to test the direct regulation of miR-214-3p on the 3'-untranslated region (UTR) of ARHGEF12. RESULTS miR-214-3p was significantly upregulated in hypoxia-treated PASMCs. Knockdown of miR-214-3p significantly attenuated hypoxia-induced proliferation and migration in PASMCs and promoted apoptosis, whereas this effect was aggravated by overexpression of miR-214-3p. In addition, dual-luciferase reporter assay demonstrated that ARHGEF12 is a direct target gene of miR-214-3p. The protein levels of ARHGEF12 were downregulated after knockdown of miR-214-3p in PASMCs. Rescue experiment results indicated that decreased proliferation of PASMCs resulted from knockdown of miR-214-3p were partially reversed by silencing of ARHGEF12 by siRNA. Furthermore, knockdown of miR-214-3p reduced expression of c-jun and c-fos, but increased expression of caspase-3 in PASMCs under hypoxia. CONCLUSIONS In conclusion, these results indicate that miR-214-3p acts as a novel regulator of hypoxia-induced proliferation and migration by directly targeting ARHGEF12 and dysregulating c-jun and c-fos in PASMCs, and may be a potential therapeutic target for treating pulmonary hypertension.

Does magnesium-supplemented cardioplegia reduce cardiac injury? A meta-analysis of randomized controlled trials.

BACKGROUND: Magnesium is often used to supplement cardioplegic solutions during cardiopulmonary bypass due to its cardioprotective effect during ischemia and reperfusion. The aim of this meta-analysis was to evaluate the effects of magnesium-supplemented cardioplegia versus an inactive (placebo) control cardioplegia on reducing cardiac injury after cardiac arrest surgery, as found by randomized, controlled trials. METHODS: The Medline, Cochrane Library, and Chinese literature databases (CJFD, CBM, CSJD, Wanfang) were comprehensively searched for reports of randomized, controlled trials (RCTs) evaluating magnesium-supplemented cardioplegic solutions. The clinical parameters and outcomes of interest were the incidence of postoperative low cardiac output, auto-rebeating rate, ICU stay length, new onset postoperative atrial fibrillation, peak value of CK-MB (and/or cTnI), incidence of myocardial infarction, and in-hospital mortality. RESULTS: Ten trials, with a total of 1214 patients, were included. The frequency of low cardiac output, inotropic utilization, and myocardial infarction, as well as auto-rebeating rate, length of ICU stay and in-hospital mortality, were similar between the two groups. There was a marginal reduction in the incidence of new-onset postoperative atrial fibrillation in the magnesium-supplemented cardioplegia group. CONCLUSIONS: The advantage of magnesium-supplemented cardioplegia, compared with cardioplegia without magnesium, remains unconvincing based on the current evidence. The decision to add magnesium to the cardioplegic solution to a patient undergoing cardiac arrest surgery should be carefully considered.

A coumarin-based fluorescent probe for selective and sensitive detection of thiophenols and its application.

The development of probes for specific thiophenol detection is of great importance, due to the toxicity of thiophenols and their derivatives in the environment. In the present study, a novel fluorescent probe was rationally designed for detecting thiophenols via an intramolecular charge transfer mechanism. The developed probe selectively and sensitively distinguished thiophenols from aliphatic thiols. It displayed a large Stokes shift (145 nm) and >280-fold fluorescence enhancement. Moreover, the new probe not only displayed excellent cell permeability for the successful detection of thiophenol in HEK293 cells but also quantitatively measured thiophenols in water samples with good recovery (more than 90%), indicating that it has promising prospects for application for thiophenol sensing in environmental and biological sciences.

[Clinical Significance of Genetic and Molecular Changes in Primary Myeloid Sarcoma].

OBJECTIVE: To investigate the clinical significance of genetic and molecular changes in primary myeloid sarcoma (MS). METHODS: Fourteen patients with primary MS were selected in Jiading District Central Hospital Affiliated Shanghai University of Medicine & Health Sciences, The First People's Hospital of Lianyungang from September 2010 to December 2021. AML1-ETO fusion, PML-RARα fusion and CBFß breakage were detected by fluorescence in situ hybridization (FISH), and the mutations of NPM1, CEBPA, FLT3, RUNX1, ASXL1, KIT and TP53 genes were detected by new generation sequencing (NGS). RESULTS: Among 14 patients, the MS occurred in bone, breast, epididymis, lung, chest wall, cervix, small intestine, ovary, lymph nodes and central nervous system. The tumor cells expressed MPO (13 cases), CD34 (7 cases), CD43 (8 cases), CD68 (7 cases), CD99 (8 cases) and CD117 (6 cases). Cytogenetic abnormalities were observed in 4 cases, including 3 cases of AML1-ETO fusion and 1 case of CBFß breakage, while no PML-RARα fusion was detected. There were no significant differences in overall survival (OS) and leukemia-free survival (LFS) between patients with and without AML1-ETO fusion/CBFß breakage (both P >0.05). Among the 14 patients, the number of NPM1, CEBPA, FLT3-ITD, RUNX1, ASXL1, KIT and TP53 gene mutations was 5, 3, 5, 3, 2, 2, 1, respectively, of which 7 cases had at least one mutation in FLT3-ITD, RUNX1, ASXL1 and TP53 gene. The OS and LFS of patients with FLT3-ITD, RUNX1, ASXL1 or TP53 mutation were shorter than those without mutations (both P <0.01). CONCLUSION: The genetic and molecular abnormalities of primary MS can be detected by FISH and NGS techniques. FLT3-ITD, RUNX1, ASXL1 or TP53 mutation indicates a worse prognosis, but further clinical studies are needed to confirm it.

Effect of Hesperidin on Chronic Unpredictable Mild Stress-Related Depression in Rats through Gut-Brain Axis Pathway.

OBJECTIVES: To determine the pharmacological impact of hesperidin, the main component of Citri Reticulatae Pericarpium, on depressive behavior and elucidate the mechanism by which hesperidin treats depression, focusing on the gut-brain axis. METHODS: Fifty-four Sprague Dawley male rats were randomly allocated to 6 groups using a random number table, including control, model, hesperidin, probiotics, fluoxetine, and Citri Reticulatae Pericarpium groups. Except for the control group, rats in the remaining 5 groups were challenged with chronic unpredictable mild stress (CUMS) for 21 days and housed in single cages. The sucrose preference test (SPT), immobility time in the forced swim test (FST), and number in the open field test (OFT) were performed to measure the behavioral changes in the rats. Enzyme-linked immunosorbent assay was used to determine the levels of 5-hydroxytryptamine (5-HT) and brain-derived neurotrophic factor (BDNF) in brain tissue, and the histopathology was performed to evaluate the changes of colon tissue, together with sequencing of the V3-V4 regions of 16S rRNA gene on feces to explore the changes of intestinal flora in the rats. RESULTS: Compared to the control group, the rats in the model group showed notable reductions in body weight, SPF, and number in OFT (P<0.01). Hesperidin was found to ameliorate depression induced by CUMS, as seen by improvements in body weight, SPT, immobility time in FST, and number in OFT (P<0.05 or P<0.01). Regarding neurotransmitters, it was found that at a dose of 50 mg/kg hesperidin treatment upregulated the levels of 5-HT and BDNF in depressed rats (P<0.05). Compared to the control group, the colon tissue of the model group exhibited greater inflammatory cell infiltration, with markedly reduced numbers of goblet cells and crypts and were significantly improved following treatment with hesperidin. Simultaneously, the administration of hesperidin demonstrated a positive impact on the gut microbiome of rats treated with CUMS, such as Shannon index increased and Simpson index decreased (P<0.01), while the abundance of Pseudomonadota and Bacteroidota increased in the hesperidin-treated group (P<0.05). CONCLUSION: The mechanism responsible for the beneficial effects of hesperidin on depressive behavior in rats may be related to inhibition of the expressions of BDNF and 5-HT and preservation of the gut microbiota.

Regulatory effect of Yinchenhao decoction on bile acid metabolism to improve the inflammatory microenvironment of hepatocellular carcinoma in mice.

Hepatocellular carcinoma (HCC) is a malignant tumor with extremely high mortality. The tumor microenvironment is the "soil" of its occurrence and development, and the inflammatory microenvironment is an important part of the "soil". Bile acid is closely related to the occurrence of HCC. Bile acid metabolism disorder is not only directly involved in the occurrence and development of HCC but also affects the inflammatory microenvironment of HCC. Yinchenhao decoction, a traditional Chinese medicine formula, can regulate bile acid metabolism and may affect the inflammatory microenvironment of HCC. To determine the effect of Yinchenhao decoction on bile acid metabolism in mice with HCC and to explore the possible mechanism by which Yinchenhao decoction improves the inflammatory microenvironment of HCC by regulating bile acid metabolism, we established mice model of orthotopic transplantation of hepatocellular carcinoma. These mice were treated with three doses of Yinchenhao decoction, then liver samples were collected and tested. Yinchenhao decoction can regulate the disorder of bile acid metabolism in liver cancer mice. Besides, it can improve inflammatory reactions, reduce hepatocyte degeneration and necrosis, and even reduce liver weight and the liver index. Taurochenodeoxycholic acid, hyodeoxycholic acid, and taurohyodeoxycholic acid are important molecules in the regulation of the liver inflammatory microenvironment, laying a foundation for the regulation of the liver tumor inflammatory microenvironment based on bile acids. Yinchenhao decoction may improve the inflammatory microenvironment of mice with HCC by ameliorating hepatic bile acid metabolism.

Overexpression of HSPA2 is correlated with poor prognosis in esophageal squamous cell carcinoma.

BACKGROUND: Heat shock-related 70 kDa protein 2 (HSPA2) has been identified as a potential cancer-promoting protein expressed at abnormal levels in a subset of cancers. However, its important role in esophageal squamous cell carcinoma (ESCC) is hardly known by people. The purpose of this study is to assess HSPA2 expression and to explore its role in ESCC. METHODS: Thirty ESCC samples, paired adjacent non-cancerous tissues and normal esophageal tissues, were collected for HSPA2 detection by quantitative RT-PCR (qRT-PCR) and western blotting. Additionally, the expression of HSPA2 in ESCC and adjacent non-cancerous tissues from 120 patients was analyzed by immunohistochemistry, and correlated with clinicopathological parameters and patients' outcome. RESULTS: HSPA2 mRNA and protein were overexpressed in ESCC tissues. Overexpression of HSPA2 was significantly associated with primary tumor, TNM stage, lymph node metastases and recurrence, respectively (all, P <0.05). Kaplan-Meier curves showed that elevated HSPA2 expression was associated with shorter disease-free survival and overall survival in ESCC patients. Cox multivariate regression analysis revealed that overexpression of HSPA2 was an independent prognostic factor in disease-free survival and overall survival for ESCC patients (hazard ratio was 2.115 and 2.210, respectively, P <0.05). CONCLUSIONS: Our findings demonstrate that overexpression of HSPA2 may contribute to the malignant progression of ESCC and present a novel prognostic indicator for ESCC patients.

[Relationship between serum ß2-microglobulin and lower extremity atherosclerotic occlusive disease].

OBJECTIVE: To explore the relationship between fasting serum level of ß2-microglobulin (ß2-M) and the development of lower extremity atherosclerotic occlusive disease (LEAOD). METHODS: A total of 59 LEAOD patients at our hospital from March 2011 to August 2012 were recruited into the LEAOD group while another 32 non-LEAOD patients into the control group. Their clinical profiles and the parameters of ankle brachial index (ABI),ß2-M and high-sensitivity C-reactive protein (hsCRP) were recorded and analyzed. RESULTS: The patients had higher serum levels of ß2-M (5.3 ± 3.2 vs 2.6 ± 1.3) and hsCRP (15.1 ± 14.8 vs 8.0 ± 6.7) according to the severity in the LEAOD group than those in the control group (P < 0.05).ß2-M was correlated with smoking (ß 1.248, odds ratio[OR] 0.020, 95% confidence interval [CI] 1.221-9.942), diabetes (ß 1.524,OR 4.591, 95%CI 1.493-14.118) and ABI (ß-4.091,OR 0.017, 95%CI 0.002-0.136) . The receiver operating characteristic (ROC) curve showed that ß2-M level had some value of predicting the occurrence of LEAOD (ROCAUC 0.821, 95%CI 0.731-0.912, P < 0.01). CONCLUSION: Serum level of ß2-M may play a role in pathologic process of LEAOD. And further studies are needed to validate its value as a biomarker for LEAOD.

[Surgical management and prognostic factors in patients with spinal metastatic tumors].

OBJECTIVE: To identify the survival prognostic factors and clinical outcome of the patients with spinal metastatic tumors and to discuss the surgical treatment strategy of spinal metastatic tumors. METHODS: The patients with spinal metastatic tumors who received surgeries during January 2003 to June 2012 were enrolled. The survival was analyzed by Kaplan-Meier survival curve. The prognostic factors, divided into patient-related factors, tumor-related factors and therapy-related factors, were analyzed univariately and multivariately by Cox comparative hazard model. RESULTS: There were 453 patients were enrolled in research including 263 male and 190 female patients with an average age of (56 ± 13) years (10-86 years). The median postoperative survival was 9 months. Local recurrences and peri-operative complications were found in 78 (17.2%) and 72 (15.9%) patients, respectively. Univariate analysis showed the significant prognostic factors for postoperative survival included poor preoperative general condition (χ(2) = 4.16), severe preoperative neurologic deficit(χ(2) = 10.23), not receiving bisphosphonate therapy(χ(2) = 10.47), short disease-free interval before spinal metastasis (χ(2) = 23.31), spinal metastasis as the first manifestation (χ(2) = 10.94), rapid-growth primary tumor(χ(2) = 15.45), visceral metastasis (χ(2) = 4.10), not receiving postoperative radiotherapy(χ(2) = 18.10) and not receiving post-operative sensitive systemic therapy(χ(2) = 11.20) (P < 0.05). Multivariate analysis showed the independent prognostic factors include severe preoperative neurologic deficit (P = 0.012, 95%CI: 1.11-2.30), short disease-free interval before spinal metastasis (P = 0.023, 95%CI:1.05-1.83), rapid-growth primary tumor (P = 0.000, 95%CI:1.74-3.06), visceral metastasis (P = 0.008, 95%CI: 1.08-1.68), not receiving postoperative radiotherapy (P = 0.000, 95%CI:1.38-2.35) and not receiving post-operative sensitive systemic therapy (P = 0.045, 95%CI:1.01-1.58). CONCLUSION: The prognostic factors for survival are useful for determining the indication of operation and improving survival and clinical outcome for patients with spinal metastatic tumors.

LncRNA CALML3-AS1 modulated by m6A modification induces BTNL9 methylation to drive non-small-cell lung cancer progression.

Non-small cell lung cancer (NSCLC) is a common and lethal malignancy. The carcinogenic roles of lncRNA CALML3 antisense RNA 1 (CALML3-AS1) have been documented. However, the function and potential mechanisms of CALML3-AS1 in the progression of NSCLC need to be further explored. The molecule expression was assessed by qRT-PCR and Western blot. The subcellular localization of CALML3-AS1 was observed by fluorescence in situ hybridization (FISH). The malignant behaviors of NSCLC cells were evaluated by CCK-8, colony formation, EdU, wound healing and transwell assays. In vivo xenograft tumor and liver metastatic models were established. The molecular mechanisms were investigated by RIP, RNA pull-down and ChIP assays. The methylation level was detected by MSP. Herein, we found that CALML3-AS1 was upregulated, while butyrophilin-like 9 (BTNL9) was downregulated in NSCLC. Functionally, CALML3-AS1 depletion repressed NSCLC cell malignant phenotypes, in vivo tumor growth, and liver metastasis. Mechanistically, AlkB homolog 5 (ALKBH5) enhanced CALML3-AS1 stability via N6-methyladenosine (m6A) demethylation, whereas m6A reader YTH domain-containing 2 (YTHDC2) destabilized CALML3-AS1. Moreover, CALML3-AS1 inhibited BTNL9 transcription and expression through the recruitment of Zeste homolog 2 (EZH2). Rescue experiments demonstrated that BTNL9 downregulation counteracted sh-CALML3-AS1-mediated antitumor effects on NSCLC. Taken together, CALML3-AS1 modulated by ALKBH5 and YTHDC2 in an m6A modification dependent manner drives NSCLC progression via epigenetically repressing BTNL9.

High-efficiency all fluorescence white OLEDs with high color rendering index by manipulating excitons in co-host recombination layers.

All fluorescence white organic light-emitting diodes (WOLEDs) based on thermally activated delayed fluorescence (TADF) emitters are an attractive route to realize highly efficient and high color quality white light sources. However, harvesting triplet excitons in these devices remains a formidable challenge, particularly for WOLEDs involving conventional fluorescent emitters. Herein, we report a universal design strategy based on a co-host system and a cascaded exciton transfer configuration. The co-host system furnishes a broad and charge-balanced exciton generation zone, which simultaneously endows the devices with low efficiency roll-off and good color stability. A yellow TADF layer is put forward as an intermediate sensitizer layer between the blue TADF light-emitting layer (EML) and the red fluorescence EML, which not only constructs an efficient cascaded Förster energy transfer route but also blocks the triplet exciton loss channel through Dexter energy transfer. With the proposed design strategy, three-color all fluorescence WOLEDs reach a maximum external quantum efficiency (EQE) of 22.4% with a remarkable color rendering index (CRI) of 92 and CIE coordinates of (0.37, 0.40). Detailed optical simulation confirms the high exciton utilization efficiency. Finally, by introducing an efficient blue emitter 5Cz-TRZ, a maximum EQE of 30.1% is achieved with CIE coordinates of (0.42, 0.42) and a CRI of 84 at 1000 cd m-2. These outstanding results demonstrate the great potential of all fluorescence WOLEDs in solid-state lighting and display panels.

[Male genitourinary system lymphoma: a clinicopathological analysis].

OBJECTIVE: To investigate the clinicopathological features and immunophenotypes of male genitourinary system lymphoma. METHODS: We retrospectively studied the histopathological characteristics and immunohistochemical markers of 35 cases of male genitourinary system lymphoma, and reviewed the relevant literature. RESULTS: The 35 patients of male genitourinary system lymphoma were aged from 4 to 83 (mean 56.5) years, 28 (80%) of them > or = 50 years. Twenty-eight cases (80%) involved the testis, 3 (8.6%) the prostate, 1 (2.9%) the spermatic cord, 1 the seminal vesicles, 1 the penis and 1 the epididymis. Histologically, 22 cases (62.9%) were diffused large B cell lymphoma (DLBCL), 6 (17.1%) mucosa associated lymphoid tissue (MALT) lymphoma, 4 (11.4%) Burkitt lymphoma, 2 (5.7%) peripheral T cell lymphoma, and 1 (2.9%) plasmacytoma. CONCLUSION: Male genitourinary system lymphomas are rare tumors clinically, which occur more often in the elderly. The majority of them are B cell lymphomas, of which the most common is DLBCL, followed by MALT lymphoma and Burkitt lymphoma. T cell lymphoma and plasmacytoma are rare. The diagnosis of male genitourinary system lymphoma relies on the histopathology, and immunohistochemical markers are of high value for its definite diagnosis, classification and differential diagnosis.

m6A methyltransferase METTL3-induced lncRNA SNHG17 promotes lung adenocarcinoma gefitinib resistance by epigenetically repressing LATS2 expression.

Gefitinib has been widely applied for the treatment of lung adenocarcinoma (LUAD). However, the long-term application of gefitinib usually leads to acquired drug resistance in tumour patients, resulting in clinical treatment failure. Small nucleolar host gene 17 (SNHG17) has been shown to play a regulatory role in LUAD progression. Nevertheless, the role of SNHG17 in LUAD gefitinib resistance remains elusive. The expression pattern of SNHG17 was examined in tissues and cell lines of gefitinib-sensitive and gefitinib-resistant LUAD, respectively. Gain- and loss-of-function experiments were employed to assess the biological functions of SNHG17 in cell proliferation and apoptosis, as well as aggressive phenotypes of LUAD cells. MeRIP-qPCR and colorimetric quantificational analysis were performed to detect m6A modifications and contents. Fluorescence in situ hybridisation (FISH) and subcellular fractionation analysis were used to reveal the distribution of SNHG17. RIP and ChIP assays were performed to further validate the SNHG17/EZH2/LATS2 regulatory axis. A xenograft tumour growth assay was conducted to evaluate the role of SNHG17 in LUAD gefitinib resistance in vivo. SNHG17 was upregulated in gefitinib-resistant LUAD tissues and cell lines. Functional assays showed that SNHG17 aggravated the malignant phenotypes of gefitinib-resistant LUAD cells. In addition, METTL3-mediated N6-methyladenosine modification could induce the upregulation of SNHG17by stabilising its RNA transcript. Mechanistically, SNHG17 epigenetically repressed the expression of LATS2 by recruiting EZH2 to the promoter region of LATS2. The regulatory role of the SNHG17/EZH2/LATS2 axis in LUAD gefitinib resistance was further supported in vivo. Collectively, our findings suggested that SNHG17 induced by METTL3 could promote LUAD gefitinib resistance by epigenetically repressing LATS2 expression.

[Atherosclerotic stenosis of intracranial and extracranial cerebral arteries in patients with cerebral infarction and the correlative factors].

OBJECTIVE: To investigate the distribution of atherosclerotic stenosis of cerebral arteries in Chinese patients suffered from cerebral infarction, and to determine if there are any factors correlating with intracranial and extracranial atherosclerosis. METHODS: For this study, we enrolled 428 consecutive in-patients with cerebral infarction, All patients were examined with transcranial Doppler ultrasonography (TCD) and carotid duplex ultrasound to detect atherosclerotic lesions in intracranial and extracranial cerebral arteries, some also were examined by magnetic resonance angiography (MRA) and/or digital subtraction angiography (DSA), they were all diagnosed as atherosclerotic cerebral artery stenosis. The patients were divided into three groups according to the different location of lesions, the frequency of risk factors of atherosclerosis and the demographic parameters were compared among these groups. RESULTS: 199 cases (46.5%) had only intracranial artery stenosis, 129 cases (30.1%) had only extracranial artery stenosis, 100 cases (23.4%) had both intracranial and extracranial stenosis (combine group). Compared with the other two groups, the levels of total cholesterol and low density lipoprotein-cholesterol of the intracranial artery group are both significantly higher (TC: P(1) = 0.001, P(2) = 0.000; LDL-C: P(1) = 0.004, P(2) = 0.039), the combine group had a significant higher ratio of male than that of the intracranial artery group(P = 0.003), there were no any other atherosclerosis risk factors had association with the location of cerebral artery stenosis. CONCLUSION: The occurrence of stenosis in intracranial arteries is more frequent than that in extracranial arteries in Chinese patients with cerebral infarction, and the lipid level is higher in the intracranial artery group, most risk factors of atherosclerosis may not be major determinants of location for cerebral atherosclerosis.

[Clinicopathological analysis of testicular mixed germ cell tumor].

OBJECTIVE: To investigate the clinicopathological characteristics of primary testicular mixed germ cell tumor (MGCT). METHODS: We retrospectively analyzed the clinicopathological data of 13 cases of primary testicular MGCT and reviewed other relevant literature. RESULTS: MGCT accounted for 24.1% (13/54) of all the testicular germ cell tumors diagnosed in our hospital. The patients ranged in age from 2 to 53 years, averaging at 28.3 years. All were unilateral cases, 6 in the left and 7 in the right testis, with a left/right ratio of 0.86:1. Morphologically, testicular MGCT displayed a variety of subtypes, embryonal carcinoma in 11 cases (84.6%), seminoma in 8 (61.5%), teratoma in 6 (46.2%), choriocarcinoma in 4 (30.8%) and yolk sac tumor in 4 (30.8%). Nine of the cases (69.2%) were composed of two different germ cell histological elements, 3 (23.1%) composed of three, and 1 (7.7%) composed of five. CONCLUSION: Testicular MGCT is rather rare and most commonly occurs in young men. Its biological behavior, clinical management and prognosis vary with its different histological elements. Therefore accurate pathological diagnosis is essential and immunohistochemistry plays an important role in the diagnosis and differential diagnosis of testicular MGCT.

PDGF mediates pulmonary arterial smooth muscle cell proliferation and migration by regulating NFATc2.

The reconstruction of pulmonary vascular structure caused by the proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs) is the central link in the formation of pulmonary arterial hypertension (PAH). Platelet­derived growth factor (PDGF) can regulate the proliferation and migration of PASMCs. At the same time, nuclear factor of activated T cells (NFATs) plays an important role in the development of PAH. To the best of our knowledge, there are no reports yet regarding whether PDGF regulates NFATc2 to increase the proliferation of PASMCs. The present study aimed to investigate whether PDGF affects the proliferation and migration of PASMCs by regulating NFAT, and to study the pathogenesis of PAH. PASMCs were treated with recombinant PDGF; Cell Counting Kit­8 and clone formation experiments showed that PDGF enhanced the cell viability and proliferation of PASMCs. Cell cycle distribution and molecular markers related to cell proliferation (cyclin D1, CDK4 and Proliferating Cell Nuclear Antigen) were detected by flow cytometry, and the results indicated that PDGF promoted the division of PAMSCs. The scratch migration and Transwell migration assays showed that the migratory ability of PASMCs was enhanced following PDGF treatment. Changes in NFATs (NFATc1­5) after PDGF treatment were evaluated by reverse transcription­quantitative PCR and western blotting; NFATc2 showed the most significant results. Finally, PDGF­treated cells were treated with an NFAT pathway inhibitor, cyclosporin A, or a small interfering RNA targeting NFATc2, and changes in cell proliferation and migration were evaluated to assess the role of NFATc2 in PDGF­induced cell proliferation and migration. In conclusion, PDGF may regulate PASMC proliferation and migration by regulating the expression of NFAT, further leading to the occurrence of PAH. It is proposed that NFATc2 could be used as a potential target for PAH treatment.