Budesonide-liposomes inclusion complex in thermosensitive gel alleviates DSS-induced ulcerative colitis in mice.

PURPOSE: A novel formulation for Ulcerative Colitis (UC) treatment by rectal administration with budesonide liposomes (Bud Lip) and thermosensitive gel (Gel) was developed for future clinical use. To evaluate the anti-inflammatory activity and colon mucosal protection of this novel formulation compared with the other three in mice. METHODS: Bud Lip was prepared by reverse evaporation method and then dispersed in solutions with PL407 and PL188 by a cold method. Male mice were induced to UC by dextran sulfate sodium (DSS) and were treated for 14 days by rectal administration, as follows: Bud enema (a conventional suspension formulation); Bud Lip; Bud Gel; Bud Lip-Gel; saline. And a negative control without colitis was also used. Disease activity index (DAI), and macroscopic and microscopic damage scores in colon tissues were used to evaluate the effect of therapy. The levels of IL-6 and IL-10 in serum and the concentrations of TNF-α and IL-10 and myeloperoxidase (MPO) activity in colon tissue were also introduced. RESULTS: In UC mice model, Bud Lip-Gel showed inflammation was alleviated significantly, and the treatment was highly associated with lower DAI, less macroscopic and microscopic colonic damage and downregulation of pro-inflammatory cytokines TNF-α, IL-6 and MPO. Bud Lip-Gel had advantages over Bud, Bud Lip, Bud Gel in the treatment of active UC. CONCLUSION: Novel Bud liposomes complex in thermosensitive Gel effectively mitigated symptoms, alleviated macroscopic and microscopic colon damage, and reduced inflammatory reaction in UC mice, which might be a potential strategy for UC treatment.

MiR-30c-5p mediates inflammatory responses and promotes microglia survival by targeting eIF2α during Cryptococcus neoformans infection.

Cryptococcosis is a disease predominantly caused by Cryptococcus neoformans in China and C. neoformans is the main form that causes cryptococcal meningitis. In this study, we examined the influence of MiR-30c-5p during Cryptococcus neoformans infection. microRNAs were extracted from Cerebrospinal fluid and sera of patients. To identify pathogenic microRNAs, RNASeq were performed. The results were confirmed with quantitative real-time PCR (qRT-PCR), transient transfection of siRNAs or microRNA mimics into cultured BV2 cell, flow cytometry, immunoblotting, luciferase assay and immunohistochemistry. In this study we found that miR-30c expression was downregulated and that inflammation, apoptosis, and autophagy were activated. The overexpression of miR-30c-5p significantly inhibited inflammation and autophagic activity and decreased apoptosis, and treatment with sieIF2α resulted in a significant decrease in inflammation, apoptosis. In addition, clinical samples of cerebrospinal fluid and serum of patients with cryptococcal meningitis who have undergone standard antifungal treatment showed that the expression of miR-30c-5p was increased while that of eIF2α was decreased, which was in accordance with the in vitro experiments. These studies demonstrated that miRNA-30c-5p can inhibit inflammatory, apoptotic, and autophagic activity through the eIF2α/ATF4 pathway, and it is thus a potential target for the diagnosis, treatment, and detection of cryptococcal meningitis.

Preoperative prediction of papillary thyroid microcarcinoma via multiparameter ultrasound.

Background Accurate diagnosis of papillary thyroid microcarcinoma (PTMC) is important for further management. Ultrasound (US) is the most frequently used imaging modality for PTMC. Purpose To evaluate the diagnostic value of conventional US, contrast-enhanced ultrasound (CEUS) and real-time elastography (RTE) for patients with PTMC. Material and Methods In total, 135 patients with subcentimeter thyroid nodules who underwent conventional US, CEUS, and RTE before surgery were enrolled. A multivariate logistic regression analysis was performed to assess the independent predictors of PTMC. The diagnostic performances of conventional US, CEUS, and RTE were evaluated with a receiver operating characteristic (ROC) curve analysis. Results A taller-than-wide shape was identified as the strongest predictor of PTMC (odds ratio [OR], 25.21), followed by heterogeneous enhancement (OR, 24.03), marked hypoechogenicity (OR, 21.71), poorly defined margin (OR, 5.51), strain ratio (OR, 2.59), and age (OR, 0.92; all P values < 0.05). Heterogeneous enhancement on CEUS showed the highest positive predictive value (PPV; 88.0%) and an accuracy of 83.7%. A logistic regression model was created to predict PTMC using conventional US, CEUS, and RTE. The area under the ROC curve was 0.97, with a sensitivity of 88.6% and a specificity of 94.6%. Conclusion Conventional US combined with CEUS and RTE can improve the diagnostic accuracy of PTMC.

Pharmacokinetic study of harmane and its 10 metabolites in rat after intravenous and oral administration by UPLC-ESI-MS/MS.

Context The ß-carboline alkaloid harmane is widely distributed in common foods, beverages and hallucinogenic plants. Harmane exerts potential in therapies for Alzheimer's and depression diseases. However, little information on its dynamic metabolic profiles and pharmacokinetics in vivo is currently available. Objective This study investigates the dynamic metabolic profiles and pharmacokinetic properties of harmane and its metabolites in rats in vivo. Materials and methods A highly selective, sensitive and rapid ultra-performance liquid chromatography combined with electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) method was developed and well-validated for simultaneous quantitative determination of harmane and its uncertain endogenous metabolite harmine, as well as for semiquantitative determination of 10 harmane metabolites in rats after intravenous injection and oral administration of harmane at 1.0 and 30.0 mg/kg, respectively. Results The calibration curves of harmane and harmine showed excellent linearity within the concentration range of 1-2000 ng/mL with acceptable accuracy, precision, selectivity, recovery, matrix effect and stability. Ten metabolites, including harmane but not harmine, were detected and identified after intravenous and oral administration of harmane. The absolute bioavailability of harmane following an oral dose was 19.41 ± 3.97%. According to the AUC0-t values of all the metabolites, the metabolic levels of phase II metabolites were higher than those of phase I metabolites, and the sulphation pathways were the dominant metabolic routes for harmane in both routes of administration. Discussion and conclusion The pharmacokinetic properties of harmane and its 10 metabolites in rats were determined. Sulphate conjugation was the predominant metabolic process of harmane in rats.

Interrogation of ERG gene rearrangements in prostate cancer identifies a prognostic 10-gene signature with relevant implication to patients' clinical outcome.

OBJECTIVES: ERG-gene rearrangement defines a distinct molecular subtype of PCA with potential biological and clinical implications. To identify a molecular signature reflective of the downstream effects of ERG-mediated transcriptional regulation with prognostic implication in patients with prostate cancer (PCA). MATERIAL AND METHODS: We used a singular value decomposition (SVD) bioinformatics approach to re-analyse gene expression data previously generated from 46 prostate tumours, and identified an ERG-like gene signature. The signature was validated on several patient cohorts and individual genes were correlated to ERG expression and PCA progression. RESULTS: An ERG-like 10-gene signature was identified and validated in PCA cohorts of the physician health study (p115) (n = 110) in addition to three independent public datasets, and was significantly associated with disease progression, biochemical recurrence and PCA-specific mortality. Patients with the ERG-like signature were significantly associated with disease recurrence on univariate (hazard ratio [HR] 2.6; 95% confidence interval [CI]:1.3-5.2; P = 0.004) and multivariate analysis (HR 2.3; 95% CI:1.1-4.6, P = 0.016) compared with patients without this signature. Within the group of patients with Gleason score (GS) 6 and 7 PCA, the signature added prognostic value beyond GS and identified patients at higher risk of cancer deaths more accurately than GS alone or in combination with ERG status. Protein expression of the 10 genes were significantly associated with ERG and disease progression regardless of ERG status. CONCLUSION: The characterized ERG-like signature was reflective of aggressive features of ERG-mediated transcription and was prognostically robust. The combination of this signature with clinicopathological variables should be validated prospectively to explore its clinical utility in stratifying patients with PCA and in identifying those at higher risk of metastatic and lethal disease.

Chemical fingerprint and simultaneous determination of alkaloids and flavonoids in aerial parts of genus Peganum indigenous to China based on HPLC-UV: application of analysis on secondary metabolites accumulation.

The aerial parts of genus Peganum are officially used in traditional Chinese medicine. The paper aims to establish a high-performance liquid chromatography (HPLC) method for fingerprint analysis and simultaneous determination of three alkaloids and two flavonoids in aerial parts of genus Peganum, and to analyze accumulative difference of secondary metabolites in inter-species, individuals of plants, inter-/intra-population and from different growing seasons. HPLC analysis was performed on a C18 column with gradient elution using 0.1% trifloroacetic acid and acetonitrile as mobile phase and detected at 265 nm, by conventional methodology validation. For fingerprint analysis, the RSDs of relative retention time and relative peak area of the characteristic peaks were within 0.07-0.78 and 0.94-9.09%, respectively. For simultaneous determination of vasicine, harmaline, harmine, deacetylpeganetin and peganetin, all calibration curves showed good linearity (r > 0.9990) within the test range. The relative standard deviations of precision, repeatability and stability test did not exceed 2.37, 2.68 and 2.67%, respectively. The average recoveries for the five analytes were between 96.47 and 101.20%. HPLC fingerprints play a minor role in authenticating and differentiating the herbs of different species of genus Peganum. However, the secondary metabolites levels of alkaloids and flavonoids in aerial parts of genus Peganum rely on species-, habitat-, and growth season-dependent accumulation.

Out of the Qinghai-Tibet Plateau: evidence for the origin and dispersal of Eurasian temperate plants from a phylogeographic study of Hippophaë rhamnoides (Elaeagnaceae).

Numerous temperate plants now distributed across Eurasia are hypothesized to have originated and migrated from the Qinghai-Tibet Plateau (QTP) and adjacent regions. However, this hypothesis has never been tested through a phylogeographic analysis of a widely distributed species. Here, we use Hippophaë rhamnoides as a model to test this hypothesis. We collected 635 individuals from 63 populations of the nine subspecies of H. rhamnoides. We sequenced two maternally inherited chloroplast (cp) DNA fragments and also the bi-paternally inherited nuclear ribosomal ITS. We recovered five major clades in phylogenetic trees constructed from cpDNA and internal transcribed spacer (ITS) sequence variation. Most sampled individuals of six subspecies that are distributed in northern China, central Asia and Asia Minor/Europe, respectively, comprised monophyletic clades (or subclades) nested within those found in the QTP. Two subspecies in the QTP were paraphyletic, while the placement of another subspecies from the Mongolian Plateau differed between the ITS and cpDNA phylogenetic trees. Our phylogeographic analyses supported an 'out-of-QTP' hypothesis for H. rhamnoides followed by allopatric divergence, hybridization and introgression. These findings highlight the complexity of intraspecific evolutions and the importance of the QTP as a center of origin for many temperate plants.

Identification of lncRNAs and Their Regulatory Relationships with mRNAs in Response to Cryptococcus neoformans Infection of THP-1 Cells.

Aims: Cryptococcosis is an invasive fungal disease that is associated with an increasing prevalence along with a very high fatality and is primarily caused by Cryptococcus. However, its mechanism to cause pathogenicity is not yet completely understood. In this study, we aim to screen the lncRNA markers in human monocytic (THP-1) cells infected by Cryptococcus neoformans (C. neoformans) through high-throughput sequencing technology and to explore its effects on biological functions. Methods: We initially conducted an lncRNA microarray analysis of the THP-1 cells infected by C. neoformans and normal THP-1 cells. Based upon these data, RT-qPCR was used to verify the expressions of the selected lncRNAs and mRNAs. We then performed functional and pathway enrichment analyses. Lastly, target prediction was performed by using the lncRNA target tool which was based on the differentially expressed lncRNAs. Results: We determined 81 upregulated and 96 downregulated lncRNAs using microarray. In addition, the profiling data showed 42 upregulated and 57 downregulated genes and discovered that neuroactive ligand-receptor interaction, tyrosine metabolism, and phenylalanine metabolism are extremely impaired in the regulation of C. neoformans infection. GO enrichment analysis of the 99 differentially expressed mRNAs exhibited that these modules showed different signaling pathways and biological mechanisms like protein binding and metal ion binding. Moreover, lncRNAs and mRNAs were analyzed for their coexpression relations. A qRT-PCR analysis confirmed that the expression of the top 10 differently expressed mRNA and lincRNA. The expressions of the lncRNAs after C. neoformans infection in THP-1 cells were detected by RNA-sequence, suggesting that microarray analysis could reveal lncRNAs having functional significance that might be linked with the progression of patients. Conclusion: The current study analyzed the differential lncRNAs and mRNAs in C. neoformans infection and predicted the corresponding pathways and their correlations that can offer new potential insights into the mechanistic basis of this condition.

COVID-19 Knowledge and Pandemic-Associated Distress Among the Hospital Pharmacist Workforce in China.

BACKGROUND: The unprecedented disruption brought about by the global coronavirus disease 2019 (COVID-19) pandemic had produced tremendous influence on the practice of pharmacy. Sufficient knowledge of pharmacists was needed to deal with the epidemic situation; however, outbreak also aggravated psychological distress among health-care professionals. Therefore, this study aimed to determine knowledge about the pandemic and related factors, prevalence and factors associated with psychological distress among hospital pharmacists of Xinjiang Province, China. METHODS: An anonymous online questionnaire-based cross-sectional study was conducted by means of WeChat, a popular social media platform in China, February 23-27, 2020, during the COVID-19 outbreak. The survey questionnaire consisted of 4 parts, including informed consent section, demographic section, knowledge about COVID-19, and assessment of overall mental health through World Health Organization's Self-Reporting Questionnaire (SRQ-20). A score of 8 or above on SRQ-20 was used as cutoff to classify the participant as in psychological distress. SRQ-20 score and related knowledge score were used as dependent variables, demographic characteristics (such as gender, age, monthly income, etc.) were used as independent variables, and univariate binary logistic regression was used to screen out the variables with P < 0.05. Then, the filtered variables were used as independent variables, and multivariate logistic regression models were used to analyze associations with sufficient knowledge of COVID-19 and psychological distress. RESULTS: A total of 365 pharmacists participated in the survey, fewer than half (35.1%; n = 128) of pharmacists attained a score of 6 or greater (out of 10) in overall disease knowledge, and most were able to select effective disinfectants and isolation or discharge criteria. In the multivariable model, age ages 31-40 (odds ratio [OR] = 3.25; P < 0.05), ages 41-50 (OR = 2.96; P < 0.05) versus >50 (referent); primary place of practice in hospitals: drug supply (OR = 4.00; P < 0.01), inpatient pharmacy (OR = 2.06, P < 0.01), clinical pharmacy (OR = 2.17, P < 0.05) versus outpatient pharmacy (referent); monthly income Renminbi (RMB, China's legal currency) 5000-10,000 (OR = 1.77; P < 0.05) versus < 5000 (referent); contact with COVID-19 patients or suspected cases (OR = 2.27; P < 0.01); access to COVID-19 knowledge remote work+ on-site work (OR = 6.07; P < 0.05), single on-site work (OR = 6.90; P < 0.01) versus remote work (referent) were related to better knowledge of COVID-19. Research found that 18.4% of pharmacists surveyed met the SRQ-20 threshold for distress. Self-reported history of mental illness (OR = 3.56; P < 0.05) and working and living in hospital versus delay in work resumption (OR = 2.87; P < 0.01) were found to be risk factors of psychological distress. CONCLUSIONS: Further training of COVID-19 knowledge was required for pharmacists. As specific pharmacist groups were prone to psychological distress, it was important for individual hospitals and government to consider and identify pharmacists' needs and take steps to meet their needs with regard to pandemic and other work-related distress.

miR­4792 regulates inflammatory responses in Cryptococcus neoformans­infected microglia.

Investigating the factors that influence the inflammatory response of microglial cells is crucial for understanding the pathogenesis of cryptococcal meningitis (CM). MicroRNAs (miRNAs/miRs) play an important role in inducing host defenses and activating the immune response during microbial infection; however, the regulatory mechanisms of miRNAs in cryptococcal meningitis remain poorly defined. In a previous study, the authors assessed the miRNA profiles of THP­1 (human acute monocytic leukemia cells) cells following Cryptococcus neoformans (C. neoformans) infection. In the present study, it was found that miR­4792 expression was downregulated in BV2 cells infected with C. neoformans, whilst that of its target gene, epidermal growth factor receptor (EGFR), was upregulated. Infected cells in which miR­4792 was overexpressed exhibited a decreased EGFR transcript expression, reduced mitogen­activated protein kinase (MAPK) signaling and a decreased secretion of inflammatory cytokines. In addition, following antifungal treatment in patients with cryptococcal meningitis, the levels of miR­4792 in the cerebrospinal fluid significantly increased, whilst the expression of EGFR significantly decreased. In addition, receiver operator characteristic analysis revealed miR­4792 (AUCROC=0.75) and EGFR (AUCROC=0.79) as potential diagnostic markers in patients with cryptococcal meningitis.

Uncovering the potential differentially expressed miRNAs as diagnostic biomarkers for hepatocellular carcinoma based on machine learning in The Cancer Genome Atlas database.

The present study aimed to identify novel diagnostic differentially expressed microRNAs (miRNAs/miRs) in order to understand the molecular mechanisms underlying hepatocellular carcinoma. The expression data of miRNA and mRNA were downloaded for differential expression analysis. Optimal diagnostic differentially expressed miRNA biomarkers were identified via a random forest algorithm. Classification models were established to distinguish patients with hepatocellular carcinoma and normal individuals. A regulatory network between optimal diagnostic differentially expressed miRNA and differentially expressed mRNAs was then constructed. The GSE63046 dataset and in vitro experiments were used to validate the expression of the optimal diagnostic differentially expressed miRNAs identified. In addition, diagnostic and prognostic analyses of optimal diagnostic differentially expressed miRNAs were performed. In total, 14 differentially expressed miRNAs (all upregulated) and 2,982 differentially expressed mRNAs (1,989 upregulated and 993 downregulated) were identified. hsa­miR­10b­5p, hsa­miR­10b­3p, hsa­miR­224­5p, hsa­miR­183­5p and hsa­miR­182­5p were considered as the optimal diagnostic biomarkers for hepatocellular carcinoma. The mRNAs targeted by these five miRNAs included secreted frizzled related protein 1 (SFRP1), endothelin receptor type B (EDNRB), nuclear receptor subfamily 4 group A member 3 (NR4A3), four and a half LIM domains 2 (FHL2), NK3 homeobox 1 (NKX3­1), interleukin 6 signal transducer (IL6ST) and forkhead box O1 (FOXO1). 'Bile acid biosynthesis and cholesterol' was the most enriched signaling pathways of these target mRNAs. The expression validation of the five miRNAs was consistent with the present bioinformatics analysis. Notably, hsa­miR­10b­5p and hsa­miR­10b­3p had a significant prognosis value for patients with hepatocellular carcinoma. In conclusion, the five differentially expressed miRNAs may be considered as diagnostic biomarkers for patients with hepatocellular carcinoma. In addition, the differential expression levels of the targets of these five mRNAs, including SFRP1, EDNRB, NR4A3, FHL2, NKX3­1, IL6ST and FOXO1, may be involved in hepatocellular carcinoma tumorigenesis.

Repeated range expansion and glacial endurance of Potentilla glabra (Rosaceae) in the Qinghai-Tibetan plateau.

To date, little is still known about how alpine species occurring in the Qinghai-Tibetan Plateau (QTP) responded to past climatic oscillations. Here, by using variations of the chloroplast trnT-L, we examined the genetic distribution pattern of 101 individuals of Potentilla glabra, comprising both the interior QTP and the plateau edge. Phylogenetic and network analyses of 31 recovered haplotypes identified three tentative clades (A, B and C). Analysis of molecular variance (amova) revealed that most of the genetic variability was found within populations (0.693), while differentiations between populations were obviously distinct (F(st)= 0.307). Two independent range expansions within clades A and B occurring at approximately 316 and 201 thousand years ago (kya) were recovered from the hierarchical mismatch analysis, and these two expansions were also confirmed by Fu's F(S) values and 'g' tests. However, distant distributions of clade C and private haplotypes from clades A and B suggest that they had survived the Last Glacial Maximum (LGM) and previous glaciers in situ since their origins. Our findings based on available limited samples support that multiple refugia of a few cold-enduring species had been maintained in the QTP platform during LGM and/or previous glacial stages.

TGF-ß-mediated upregulation of brachyury contributes to constitutively up-regulated type I collagen expression in skin fibroblasts: possible role in systemic sclerosis.

BACKGROUND: Previous reports have shown that epithelial-to-mesenchymal transition (EMT) indicates the importance of transforming growth factor-ß (TGF-ß) signalling in the pathogenesis of systemic sclerosis (SSc). However, the underlying molecular mechanisms of EMT are not fully understood. OBJECTIVES: Brachyury, an evolutionarily conserved transcription factor, was recently identified as an important factor that promotes EMT in human carcinoma cell lines. However, there is no evidence indicating that brachyury is involved in EMT in SSc. MATERIALS AND METHODS: The expression of brachyury and collagen was investigated in cultures of dermal fibroblasts and skin sections derived from SSc patients and healthy controls. Brachyury and collagen expression were determined by immunohistochemistry and immunoblotting, respectively, and mRNA for both was analysed using real-time PCR. RESULTS: Brachyury was overexpressed in SSc dermal fibroblasts both in vivo and in vitro, and this overexpression was inhibited by TGF-ß1 inhibitor. Brachyury siRNA reduced mRNA and protein expression levels of type I collagen in normal and SSc dermal fibroblasts, but did not decrease the levels of major disease-related cytokines. Furthermore, brachyury levels were significantly increased in skin samples of SSc patients relative to healthy controls. CONCLUSIONS: The up-regulation of brachyury in response to activated endogenous TGF-ß signalling may play a role in constitutive up-regulation of collagen in SSc fibroblasts. Further studies assessing the regulatory mechanism of tissue fibrosis induced by brachyury in SSc skin may lead to a better understanding of the pathogenesis, new diagnostic methods, and new therapeutic approaches using siRNAs.

Expression of survivin in invasive pituitary adenoma.

OBJECTIVE: To investigate the relationship between survivin expression and invasiveness of pituitary adenoma. METHODS: A total of 66 patients, on whom trans sphenoidal surgery had been performed between July 2006 and March 2008, were enrolled in our study at the Department of Neurosurgery in Shandong Provincial Hospital and Jinan Central Hospital, Shandong, P. R. China. All patients were divided into the invasion group (n=39), and the non-invasion group (n=27) by assessment of preoperative MRI and intraoperative inspection. Survivin expression was determined by immunohistochemistry. Statistical analysis of survivin expression between the 2 sample groups was accomplished using the chi-square test. RESULTS: Survivin was expressed in 46 (69.7%) of the investigated pituitary adenomas. For invasive pituitary adenoma, survivin staining was positive in 35 (89.7%), only 11 (40.7%) specimens were positive in noninvasive tumors. The chi-square test demonstrated a statistically significant difference in survivin expression between invasive and noninvasive pituitary adenoma (chi2=14.309, p=0.0002). CONCLUSION: Survivin was highly associated with invasive pituitary adenoma, it is likely to serve as a useful tool for confirmation of invasive pituitary adenoma and the gene could be an effective target for pituitary adenoma gene therapy.

Expression and significance of glucocorticoid receptor alpha in meningiomas.

OBJECTIVE: To explore the expression of the glucocorticoid receptor alpha (GRalpha) and its significance in the occurrence and progress of meningiomas. MATERIALS AND METHODS: By the use of flow cytometry, the proliferative index (PI), S-phase fraction (SPF) and DNA ploidy were detected to evaluate the proliferation of tumor cells in 58 meningioma specimens. The expressions of GRalpha in all meningiomas and seven normal dura samples were studied by means of reverse transcription-polymerase chain reaction to compare the difference in GRalpha between meningiomas and normal dura. The relation between GRalpha and histological grades, PI, SPF and DNA ploidy were also analyzed. RESULTS: The mean PI was 9.32%+/-4.41% while the mean SPF was 2.79%+/-2.43% in 58 meningioma specimens. DNA was diploid in 51 cases and aneuploid in the remaining seven cases, with the aneuploid rate being 12.1%. Nine of 58 meningiomas were GRalpha- negative and the rest were GRalpha-positive with a GRalpha-positive rate of 84.5%. The GRalpha-positive meningiomas included 13 '+', 15 '++', 9 '+++' and 12 '++++'. GRalpha was negative in normal dura samples. The GRalpha-positive rate of meningiomas was significantly greater than that of normal dura. There were no significant differences in PI and SPF among GRalpha-negative, GRalpha-weak positive and GRalpha-strong positive meningiomas. The difference in aneuploid rate between GRalpha-positive and GRalpha-negative meningiomas was also not significant. CONCLUSION: GRalpha is of significance in meningiomas, which are a target tissue for glucocorticoid. However, GRalpha's expression had no obvious effect on the proliferative activity of meningiomas, so it may not be a major control of this process.

[Expression of annexin I in different histological types of carcinomas].

OBJECTIVE: To investigate the expression of annexin I in esophageal squamous cell carcinoma (SCC) and carcinomas of other histological types in order to analyze the correlation between the expression of annexin I and carcinogenesis. METHODS: First, a set of tissue microarray was established, which consisted of SCC from the esophagus (208 cases), lung, larynx, cervix, and external genital organs; adenocarcinomas from the lung, stomach, colon and rectum, liver, pancreas, breast, thyroid and kidney with 30 cases in each group, meanwhile, the corresponding normal tissue was also obtained for control. Immunohistochemistry was used to detect the expression of annexin I in different types of carcinomas and the corresponding normal controls from different organs. The correlation between the expression of annexin I and the clinicopathological feature was analyzed and compared, which included age, gender, differentiation grade and lymph node metastasis. RESULTS: It was found that the expression of annexin I was decreased in esophageal SCC, when compared with normal esophageal squamous epithelia (P < 0.001), the similarity was also found in SCC of the lung, larynx and cervix. However, though negative in normal epidermis, annexin I expression was detected in some cases with SCC from external genital organs. Annexin I was found to be overexpressed in adenocarcinomas of the lung, stomach, colon and rectum, liver, pancreas, breast, thyroid and kidney, particularly very strong expression of annexin I was seen in lung adenocarcinoma, uterine endometrioid adenocarcinoma and ovarian serous adenocarcinoma. Interestingly, it was found to be positive in all thyroid papillary carcinomas, but negative in all normal thyroid glands. However, annexin I expression was found to be negative in all hepatocellular carcinoma and normal hepatocytes; and it was only detected in myoepithelium of normal breast tissue, but not in ductal luminal cells, and rarely in infiltrating ductal adenocarcinoma. In SCC, annexin I expression was stronger in well differentiated ones than that in the poorly differentiated ones. However, contrasting with SCC, in the adenocarcinomas from different organs, annexin I expression was much stronger in poorly differentiated ones than that in the well differentiate ones, especially in the adenocarcinomas from stomach, colon and rectum, pancreas, ovarian and kidney. CONCLUSION: Annexin I expression is quite different among different types of carcinomas, and is correlated with histopathological type and differentiation grade. Further study is needed to investigate its role in the carcinogenesis.

Prognostic significance of the International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society classification of stage I lung adenocarcinoma: A retrospective study based on analysis of 110 Chinese patients.

BACKGROUND: The aim of this study was to investigate the relationship between predominant subtype, classification, and prognosis in Chinese stage I lung adenocarcinoma patients according to the International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society (IASLC/ATS/ERS) International Multidisciplinary Lung Adenocarcinoma Classification. METHODS: Between 2000 and 2010, 110 patients with stage I lung adenocarcinoma underwent surgery at Xuanwu Hospital. Two pathologists independently reclassified all resected specimens according to the IASLC/ATS/ERS classification. Survival curves were plotted using the Kaplan-Meier method. The Cox proportional hazard model was used for multivariate analysis. RESULTS: There were no cases of adenocarcinoma in situ, and three cases of minimally invasive adenocarcinoma. There were 107 cases of invasive adenocarcinoma: 12 lepidic, 32 acinar, 30 papillary, 18 micropapillary, and 15 solid predominant subtypes. Patients with micropapillary and solid predominant tumors had significantly poorer disease-free survival compared to those with other subtypes of predominant tumors (P = 0.021). Multivariate analysis revealed that the new classification (P = 0.003) and T stage (P = 0.034) were independent predictors of disease-free and overall survival, respectively. CONCLUSION: The predominant subtype in the primary tumor was associated with prognosis in resected stage I lung adenocarcinoma.

Interspecies metabolic diversity of harmaline and harmine in in vitro 11 mammalian liver microsomes.

The ß-carboline alkaloids harmaline and harmine are widely present in hallucinogenic plants with great potential for treating depression, Parkinson's disease, and Alzheimer's disease. The present study was to elucidate metabolic difference of harmaline and harmine in 11 mammalian liver microsomes in order to quantitate species-specific metabolic profiles. Using the probe substrate reaction, the enzymatic activities for 8 CYP450 isozymes of 11 liver microsomes were characterized. Combining ultra performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-Q/TOF-MS) and ultra performance liquid chromatography combined with electrospray ionization quadrupole tandem mass spectrometry (UPLC-ESI-MS/MS) methods, 18 metabolites for harmaline and 11 for harmine were identified. The metabolism patterns differences of them presented discrepancy in the quality and quantity of metabolites. It was notable that O-sulfate conjugation was detected in all species except sheep. The intrinsic clearance CLint, LM values for the metabolites harmine and harmol in rabbits (37.5 and 42.4 µL/min/mg) were higher than those in other animals, while dogs (16.2 and 16.7 µL/min/mg) and humans (16.0 and 16.3 µL/min/mg) exhibited similar in vitro metabolic clearance. These observations suggested that harmaline and harmine were rapidly metabolized in liver microsomes of rat, mouse, and rabbit; moderately metabolized in human and dog; while weakly metabolized in sheep. Comprehensive analysis of the metabolism indicated that dogs and humans showed considerable similarity in the elimination of parent drugs, metabolic profiles, and catalytic processes. To summarize, these findings illustrated that in vitro studies of harmaline and harmine metabolic profiles in different species are helpful for the proper selection and interpretation of animal models for pharmacological and toxicological evaluation, and will ultimately provide useful guidance for the development of ß-carboline alkaloids. Copyright © 2016 John Wiley & Sons Ltd.

Sesamol attenuates oxidative stress, apoptosis and inflammation in focal cerebral ischemia/reperfusion injury.

The aim of the present study was to evaluate the therapeutic potential of sesamol treatment on focal ischemia/reperfusion (I/R) injury in the rat brain. The results demonstrated that pretreatment with sesamol seven days prior to focal cerebral I/R injury had significant positive effects, including improvements in neurological deficits (P<0.05), and a reduction in malondialdehyde content and elevation of antioxidant levels (superoxide dismutase, glutathione and glutatione peroxidase; both P<0.05). Furthermore, levels of B cell lymphoma-2 (Bcl-2)-associated X protein and caspase-3 were significantly downregulated, whereas the level of Bcl-2 was effectively increased. Conversely, the mRNA expression of proinflammatory cytokines were significantly reduced in focal cerebral I/R injury rats upon sesamol intervention. Therefore, the beneficial effects of sesamol on cerebral I/R injury may be due to the reduction of oxidative stress, inhibition of apoptosis and inflammation. The findings of the present study suggest that sesamol supplementation may serve as potent adjuvant in the treatment of focal cerebral ischemia/reperfusion injury due to its neuroprotective effects.