Mitochondrial targeted modification and anticancer mechanism of natural product ergosterol peroxide.

Ergosterol peroxide (EP) isolated from the edible medicinal fungus Pleurotus ferulae has a wide range of anti-tumor activity, but poor water solubility and low bioavailability limit further application. In this study, EP was structurally modified using triphenylphosphine (TPP+), which combines mitochondrial targeting, amphiphilicity, and cytotoxicity. A series of TPP+-conjugated ergosterol peroxide derivatives (TEn) with different length linker arms were synthesized. The structure-activity relationship showed that the anticancer activity of TEn gradually decreased with the elongation of the linker arm. The compound TE3 has the optimal and broadest spectrum of antitumor effects. It mainly through targeting mitochondria, inducing ROS production, disrupting mitochondrial function, and activating mitochondria apoptosis pathway to exert anti-cervical cancer activity. Among them, TPP+ only acted as a mitochondrial targeting group, while EP containing peroxide bridge structure served as an active group to induce ROS. In vivo experiments have shown that TE3 has better anti-cervical cancer activity and safety than the first-line anticancer drug cisplatin, and can activate the immune response in mice. Although TE3 exhibits some acute toxicity, it is not significant at therapeutic doses. Therefore, TE3 has the potential for further development as an anti-cervical cancer drug.

Myriocin enhances the clearance of M. tuberculosis by macrophages through the activation of PLIN2.

Myriocin is an inhibitor of de novo synthesis of sphingolipids and ceramides. In this research, we showed myriocin could significantly reduce Mtb burden and histopathological inflammation in mice. However, the underlying mechanism remains unclear. RNA-seq analysis revealed a significant increase in gene expression of PLIN2/CD36/CERT1 after myriocin treatment. The reduced bactericidal burden was only reversed after silencing the lipid droplets (LDs) surface protein PLIN2. This suggests that myriocin enhances the ability of macrophages to clear Mtb depending on the PLIN2 gene, which is part of the PPARγ pathway. Indeed, we observed a significant increase in the number of LDs following myriocin treatment.IMPORTANCEMycobacterium tuberculosis has the ability to reprogram host cell lipid metabolism and alter the antimicrobial functions of infected macrophages. The sphingolipids, such as ceramides, are the primary host lipids utilized by the bacteria, making the sphingomyelinase/ceramide system critical in Mtb infections. Surprisingly, the antimicrobial effect of myriocin was found to be independent of its role in reducing ceramides, but instead, it depends on the lipid droplets surface protein PLIN2. Our findings provide a novel mechanism for how myriocin enhances Mtb clearance in macrophages.

Progesterone receptor impairs immune respond and down-regulates sensitivity to anti-LAG3 in breast cancer.

BACKGROUND: Progesterone receptor (PR) serves as a crucial prognostic and predictive marker in breast cancer. Nonetheless, the interplay between PR and the tumor immune microenvironment remains inadequately understood. This investigation employs bioinformatics analyses, mouse models, and clinical specimens to elucidate the impact of PR on immune microenvironment and identify potential targets for immunotherapy, furnishing valuable guidance for clinical practice. METHODS: Analysis of immune infiltration score by Xcell between PR-positive and PR-negative breast cancer tumors. Construction of overexpression mouse progesterone receptor (mPgr) EMT-6 cell was to explore the tumor immune microenvironment. Furthermore, anti- Lymphocyte-activation gene 3 (LAG3) therapy aimed to investigate whether PR could influence the effectiveness of immune treatments. RESULTS: Overexpression mPgr inhibited tumor growth in vitro, but promoted tumor growth in Balb/c mouse. Flow cytometry showed that the proportion and cytotoxicity of CD8+T cells in tumor of overexpressing mPgr group were significantly reduced. The significant reduction in overexpressing mPgr group was found in the proportions of LAG3+CD8+ T cells and LAG3+ Treg T cells. Anti-LAG3 treatment resulted in reduced tumor growth in EV group mouse rather than in overexpressing mPgr group. Patents derived tumor fragment (PDTF) also showed higher anti-tumor ability of CD3+T cell in patents' tumor with PR <20% after anti-human LAG3 treatment in vitro. CONCLUSIONS: The mPgr promotes tumor growth by downregulating the infiltration and function of cytotoxic cell. LAG3 may be a target of ER-positive breast cancer immunotherapy. The high expression of PR hinders the sensitivity to anti-LAG3 treatment.

Clinical analysis and methodological evaluation of syphilis infection in patients in a first-class tertiary hospital in Suzhou, China.

OBJECTIVE: To explore the distribution and epidemiological characteristics of patients with syphilis in a first-class tertiary hospital and to evaluate the coincidence rate between chemiluminescence immunoassay (CLIA) and Treponema pallidum particle agglutination assay (TPPA). METHODS: The medical records of 247,501 outpatients and inpatients were retrospectively analyzed. TPPA was used to verify positive and suspected cases, and the coincidence rate between CLIA and TPPA was evaluated. Receiver operating characteristic (ROC) curve was used to determine optimal diagnostic thresholds. RESULTS: Of the 247,501 serum samples, 5,173 were detected positive for syphilis using CLIA, with a detection rate of 2.09% and a men-to-women ratio of 1.39. The chi-square test showed that sex and age were both factors that affected the detection rate (χ2=229.51, P < 0.0001). In addition, urology, orthopedics, cardiology, general surgery, gastroenterology, and gynecology represented the top six departments with the highest numbers of positive cases. Comparative analysis showed that the overall coincidence rate between CLIA and TPPA was 80.24%. Analysis of the ROC curve showed that the area under the curve (AUC) was 0.936 (95% confidence interval [CI]: 0.929-0.942, P < 0.0001) using sample/cut-off value (S/CO) as a diagnostic indicator. The results showed that an S/CO value of 3.945 was the best diagnostic value for the CLIA method, with a diagnostic specificity of 93.64% and a sensitivity of 81.90%. CONCLUSIONS: Syphilis is widely distributed in various hospital departments and primarily affects middle-aged and older individuals. For cases that have been initially screened as positive or suspicious, TPPA and other tests should be used for verification to avoid misdiagnosis and missed diagnosis.

Mycobacterium tuberculosis-Induced Prostaglandin J2 and 15-Deoxy-Prostaglandin J2 Inhibit Inflammatory Signals in Human M1 Macrophages via a Negative Feedback Loop.

Tuberculosis caused by Mycobacterium tuberculosis is a leading cause of death globally and a major health concern. In humans, macrophages are the first line invaded by M. tuberculosis. Upon infection, macrophages upregulate cyclooxygenase-2 (COX-2) expression and consequently elevate the formation of PGs, including PGE2 and PGD2. Although the role of proinflammatory PGE2 in M. tuberculosis infection has been reported, the roles of PGJ2 and 15-deoxy-PGJ2 (collectively named J2-PGs), the metabolites of PGD2 with anti-inflammatory features, remain elusive. In this study, we show that M. tuberculosis (H37Rv strain)-conditioned medium stimulates human monocyte-derived macrophages (MDMs) to elevate COX-2 expression along with robust generation of PGJ2, exceeding PGD2 formation, and to a minor extent also of 15-deoxy-PGJ2. Of interest, in M1-MDM phenotypes, PGJ2 and 15-deoxy-PGJ2 decreased M. tuberculosis (H37Rv strain)-conditioned medium-induced COX-2 expression and related PG formation by a negative feedback loop. Moreover, these J2-PGs downregulated the expression of the proinflammatory cytokines IL-6, IL-1ß, and IFN-γ, but elevated the anti-inflammatory cytokine IL-10 and the M2 markers arginase-1 and CD163. These anti-inflammatory effects of J2-PGs in M1-MDM correlated with impaired activation of TGF-ß-activated kinase 1/NF-κB/MAPK pathways. Finally, we found that J2-PGs regulate COX-2 expression, at least partially, via PGD2 receptor (DP1) and chemoattractant receptor homologue expressed on Th2 cells/DP2 receptors, but independent of the J2-PG receptor peroxisome proliferator-activated receptor-γ. Together, our findings reveal that M. tuberculosis induces COX-2 expression in human M1-MDMs, along with robust formation of J2-PGs that mediates anti-inflammatory effects via a negative feedback loop.

Construction and validation of nomograms to reduce completion thyroidectomy by predicting lymph node metastasis in low-risk papillary thyroid carcinoma.

CONTEXT: More than 5 central lymph nodes metastases (CLNM) or lateral lymph node metastasis (LLNM) indicates a higher risk of recurrence in low-risk papillary thyroid carcinoma (PTC) and may lead to completion thyroidectomy (CTx) in patients initially undergoing lobectomy. OBJECTIVE: To screen potentially high-risk patients from low-risk patients by using preoperative and intraoperative clinicopathological features to predict lymph node status. METHODS: A retrospective analysis of 8301 PTC patients in Wuhan Union Hospital database (2009-2021) was performed according to the 2015 American Thyroid Association (ATA) and 2021 National Comprehensive Cancer Network (NCCN) guidelines, respectively. Logistic regression and best subsets regression were used to identify risk factors. Nomograms were established and externally validated using the Differentiated Thyroid Cancer in China cohort. RESULTS: More than 5 CLNM or LLNM was detected in 1648 (19.9%) patients. Two predictive models containing age, gender, maximum tumor size, free thyroxine (FT4) and palpable node (all p < 0.05) were established. The nomogram based on NCCN criteria showed better discriminative power and consistency with a specificity of 0.706 and a sensitivity of 0.725, and external validation indicated that 76% of potentially high-risk patients could achieve preoperative conversion of surgical strategy. CONCLUSIONS: Models based on large cohorts with good predictive performance were constructed and validated. Preoperative low-risk (T1-2N0M0) patients with age younger than 40 years, male gender, large tumor size, low FT4 and palpable nodes may be at high risk of LLNM or more than 5 CLNM, and they should receive more aggressive initial therapy to reduce CTx.

Real-time intraoperative near-infrared autofluorescence imaging to locate the parathyroid glands: A preliminary report.

Identification and localization of parathyroid glands (PGs) remains a challenge for surgeons. The aim of this study was to evaluate the efficiency of intraoperative near-infrared autofluorescence (NIRAF) imaging to detect PGs in thyroid and parathyroid diseases. Seventy-six patients undergoing surgery for thyroid or parathyroid diseases between July 9, 2020 and August 20, 2021 were retrospectively analyzed. Intraoperative carbon nanoparticle (CN) negative imaging and handheld NIRAF imaging were successively performed for each patient. Of 206 PGs that needed to be identified for surgery, 162 were identified by NIRAF imaging, with a theoretical rate of identification of 78.64%. This was higher than the rate of identification with CN negative imaging, which was 75.73%. The number of PGs identified by NIRAF imaging and CN negative imaging did not differ significantly in either total thyroidectomy or thyroid lobectomy. In addition, the autofluorescence (AF) intensity of secondary parathyroid adenoma was weaker than that of normal PGs. NIRAF imaging is potentially a more efficient tool for identification of PGs than CN negative imaging, with a shorter learning curve and lower risk. It may not be well-suited to secondary hyperthyroidism or adenoma, but it was more efficient at identifying excised specimens than visual identification by a surgeon.

Case Report: Implantation of Dedifferentiated to Poorly Differentiated Thyroid Carcinoma After Endoscopic Thyroid Surgery.

Background: Endoscopic thyroidectomy is widely accepted for its advantages. However, implant metastasis remains a significant complication of endoscopic thyroidectomy. Methods: This is the first report of breast implantation diagnosed with poorly differentiated thyroid carcinoma following endoscopic thyroidectomy. Results: We present a case of a 35-year-old woman who was initially diagnosed with a 3.0 cm conventional papillary thyroid carcinoma after endoscopic thyroidectomy via total areola. Two years later, she was reported to have recurring poorly differentiated thyroid carcinoma in the right areola. Implantation after endoscopic thyroidectomy is rare, and even rarer is dedifferentiated papillary thyroid carcinoma around the implant site. Conclusions: Stringently evaluated endoscopic surgery indications, appropriate preoperative evaluation, meticulous surgical technique, and adequate protective measures can significantly reduce the incidence of local implantation or recurrence.

Influence of progesterone receptor on metastasis and prognosis in breast cancer patients with negative HER-2.

BACKGROUND: In de novo metastatic breast cancer patients, the site of metastasis and prognosis are related to the molecular subtype of breast cancer. There are few relevant reports to explore the clinicopathological and prognostic characteristics of different single positive hormone receptor subtypes [estrogen receptor (ER)+/progesterone receptor (PR)- and ER-/PR+] of metastatic breast cancer. METHODS: Using the Surveillance, Epidemiology and End Results (SEER) database between 2010 and 2015.We analyzed the metastatic patterns and prognosis of human epidermal growth factor receptor 2 (HER-2)-negative breast cancer patients. Cox analysis was used to analyze the influence of ER+/PR- and ER-/PR+ on the prognosis of patients in different subgroups and the risk factors affecting the prognosis of patients with single hormone receptor positivity. RESULTS: We included 206,187 breast cancer patients, including 7,726 stage IV patients. The loss of ER was a protective factor against bone metastasis (P<0.001) and a risk factor for visceral metastasis (P<0.001). The ER-/PR+ subtype had a similar proportion of de novo metastatic breast cancer, and similar clinicopathological characteristics, prognosis with triple negative breast cancer (TNBC). Single PR positivity was an independent risk factor for cancer specific survival (CSS) in multi-visceral metastasis subgroup comparing to TNBC. Meanwhile, no significant difference in overall survival (OS) or breast cancer specific survival (BCSS) between ER-/PR+ and ER-/PR- patients in all breast cancer patients or in stage IV breast cancer patients. Age [hazard ratio (HR) =2.16], grade (HR =2.36), T stage (T4: HR =3.24), lymph node metastasis (>10: HR =4.33), distant metastasis (HR =4.99), and no chemotherapy or an unknown (HR =1.65) were high-risk factors but surgery (HR <0.5) were protective factors for CSS in ER-/PR+ patients. CONCLUSIONS: ER-/PR+ subtype had a high proportion of stage IV patients. Meanwhile, such subtype breast cancer had similar clinicopathological characteristics, metastatic models (prefers to visceral metastasis), similar even worse prognosis compared with TNBC.

Presentation and survival by hormonal receptor status in metaplastic breast cancer: A propensity score-matched analysis.

BACKGROUND: Metaplastic breast cancer (MBC) is a rare and aggressive form of breast cancer. The effectiveness of chemotherapy (CT) for MBC remains controversial. The present study aimed to evaluate the efficacy of CT combined hormone receptor (HR) status on MBC patients with high risk (T1-4N2-3M0 and T4N0-1M0) by propensity-score matching (PSM). METHODS: A retrospective study was performed to analyze MBC from the SEER database. Breast cancer-specific survival (BCSS) was analyzed using the Kaplan-Meier curve. Cox proportional hazard models were used to assess BCSS. PSM was used to make 1:1 case-control matching. RESULTS: This study identified 3116 patients. The median follow-up time was 44 months (range, 1-321 months). About 62.5 % of patients received CT. 23.0 % of patients were HR-positive. Recurrence risk had a significant difference between the HR-negative and HR-positive groups. In the multivariable Cox regression model, CT had no benefit for MBC patients. HR status was not associated with a better prognosis. In subgroup analysis, the Kaplan-Meier analysis showed that HR-negative MBC with intermediate-risk benefited from CT. For HR-positive MBC, patients with intermediate and high risk also benefited from CT. After PSM, neither CT nor HR status was not related to better BCSS. Moreover, the use of CT could only improve the survival of HR-positive MBC patients with high risk. CONCLUSION: PSM analysis showed that HR status was not associated with a better prognosis. CT was not a significant prognostic factor for prognosis. However, HR-positive MBC patients with high risk might benefit from CT.

Single Hormone Receptor-Positive Metaplastic Breast Cancer: Similar Outcome as Triple-Negative Subtype.

Background: Metaplastic breast cancer (MBC) is a rare and aggressive subtype of the breast. To understand the characteristics and prognosis of single hormone receptor-positive (HR+) MBC (estrogen receptor-positive [ER+]/progesterone receptor-negative [PR-] and ER-/PR+), we compared these tumors to double HR+ tumors as well as HR- tumors. Patients and Methods: The Surveillance, Epidemiology, and End Results database was used to analyze MBC between 1975 and 2016. The effect of HR status was evaluated using a multivariate Cox regression model. Results: We included 3369 patients with a median follow-up time of 42 months (range 0-322 months). In this study, 280 (8.3%) cases were double HR+ tumors, 2597 (77.1%) were double HR- tumors, and 492 (14.6%) cases were single HR+ tumors, of which 159 (4.7%) cases were ER-/PR+ tumors and 333 (9.9%) were ER+/PR- tumors. On multivariate Cox analysis, the prognosis was related to age, race/ethnicity, tumor grade, TNM stage, and surgery. HR status remained no impact on breast cancer-specific survival (BCSS). In the Kaplan-Meier curve, HR status was not associated with better BCSS or overall survival (OS). In patients without HER2 overexpression, the BCSS and OS of ER+/PR- and ER-/PR+ tumors were not significantly different from that of ER-/PR- and ER+/PR+ tumors. The difference remains no significant in patients with HER2 overexpression. Conclusions: In comparison with both ER-/PR- and ER+/PR+ tumors, we have identified clinically and biologically distinct features of single HR+ tumors. In patients with or without HER2 overexpression, the prognosis of single HR+ tumors was similar to ER-/PR- and ER+/PR+ tumors.

Metaplastic breast cancer: Treatment and prognosis by molecular subtype.

BACKGROUND: Metaplastic breast cancer (MBC) is a rare and aggressive subtype of breast. However, the effect of molecular subtype on treatment and prognosis of MBC remains unclear. PATIENTS AND METHODS: The Surveillance, Epidemiology, and End Results database was used to analyze patients with MBC between 2010 and 2016. Molecular subtype was stratified to TN group (ER and PR-/HER2-), HER2 group (ER and PR-/HER2+, ER/PR+ and HER2+), and HR group (ER/PR+ and HER2-). The breast cancer-specific survival (BCSS) differences were estimated using multivariate Cox regression model and Kaplan-Meier curves. RESULTS: We included 1665 patients with median follow-up time of 27 months (range 0-83 months). 1154 (69.3%), 65 (3.9%), and 446 (26.8%) patients presented in TN group, HER2 group, and HR group, respectively. On multivariate Cox analysis, the prognosis was related to age, tumor size, regional node metastasis, and surgery. Molecular subtype remained no impact on BCSS. Radiotherapy (RT) was associated with better prognosis. Patients cannot benefit from chemotherapy. In Kaplan-Meier curve, triple-negative (P = 0.047) and HR-positive (P = 0.006) patients receiving RT had a superior BCSS than that not RT. HER2-positive patients cannot benefit from RT. However, adjusted Kaplan-Meier survival model showed that triple-negative (P = 0.019) but not HER2-positive (P = 0.575) or HR-positive (P = 0.574) patients receiving RT had a superior BCSS than that not RT. CONCLUSIONS: Molecular subtype is not associated with the better prognosis of MBC. Patients could benefit from RT. However, triple-negative but not HR-positive or HER2-positive patients have superior survival after receiving RT.

The Effect of Post-Mastectomy Radiotherapy in Patients With Metaplastic Breast Cancer: A Propensity Score-Matched Analysis of the SEER Database.

BACKGROUND: Metaplastic breast cancer (MBC) is a rare tumor with aggressive biological behavior. This study aimed to evaluate the efficacy of post-mastectomy radiotherapy (PMRT) on patients with low-risk (T1N0M0), intermediate-risk (T1-2N1M0 and T3N0M0), and high-risk (T1-4N2-3M0 and T4N0-1M0) MBC via propensity-score matching (PSM). METHODS: We analyzed information from the Surveillance, Epidemiology, and End Results (SEER) public-use database from 1975 to 2016 for MBC incidence trends and compared overall survival (OS) and breast cancer-specific survival (BCSS) between groups of MBC women diagnosed from 2001 to 2016 using Kaplan-Meier analysis and the multivariate Cox proportional model. PSM was used to make 1:1 case-control matching. RESULTS: Joinpoint analyses identified 1984 and 2003 as the inflection points among 4,672 patients. 1,588 (42.4%) of the 3,748 patients diagnosed with MBC between 2001 and 2016 received PMRT. According to multivariate analyses, PMRT provided better OS (p < 0.001) and BCSS (p < 0.001) before PSM, and better prognosis after PSM (n = 2528) for patients receiving PMRT (n = 1264) compared to those without PMRT (OS, p < 0.001 and BCSS, p < 0.001). When stratifying the case-control matching patients into low-risk, intermediate-risk, and high-risk groups, PMRT could improve BCSS compared with that in non-PMRT patients in the high-risk groups; it also improved OS in both the intermediate- and high-risk groups. CONCLUSIONS: Per findings of the PSM analysis, PMRT could provide better BCSS in high-risk groups, and better OS in intermediate- and high-risk groups.

Combination of serum lactate dehydrogenase and sex is predictive of severe disease in patients with COVID-19.

Elevated serum lactate dehydrogenase (LDH) was commonly reported in COVID-19 patients. However, the relationship between LDH and the incidence of severe cases has not been characterized in those patients.We retrospectively analyzed the characteristics of patients from a designated isolation medical center for COVID-19 patients diagnosed from February 6 to March 1. Variables accessed within 48 hours on admission were compared between patients with and without the severe disease. Logistic model analyses were performed to examine the prognostic value of LDH for predicting severe disease.52 (28.6%) patients later developed severe disease. Comparing to non-severe cases, severe cases had a higher level of serum LDH (321.85 ±â€Š186.24 vs 647.35 ±â€Š424.26, P < .001), neutrophils (5.42 ±â€Š3.26 vs 9.19 ±â€Š6.33, P < .001), and C-reactive protein (38.63 ±â€Š43.14 vs 83.20 ±â€Š51.01, P < .001). The patients with severe disease tended to be male (44.6% vs 80.8%, P < .001), lower level of serum albumin (31.41 ±â€Š6.20 vs 27.18 ±â€Š5.74, P < .001), and SpO2 (96.30 ±â€Š2.75 vs 92.37 ±â€Š8.29, P < .001). In the multivariate analysis model, LDH and sex remained independent risk factors for severe disease. The serum LDH predicted severe cases with an area under the curve (AUC) of 0.7999. A combination of serum LDH and sex predicted severe cases with an AUC of 0.849. A combination of serum LDH accessed on admission and sex had a better predictive performance than the serum LDH (P = .0238).Serum LDH on admission combined with sex is independently associated with severe disease in COVID-19.

CPT1A regulates breast cancer-associated lymphangiogenesis via VEGF signaling.

BACKGROUND: Lymphangiogenesis is critical for metastasis of a variety of cancers, including breast cancer. CPT1A (carnitine palmitoyltransferase 1a) has been reported to play a critical role in breast cancer progress. However, the molecular mechanism remains elusive. METHODS: In order to investigate the role of CPT1A in HDLEC cells, short hairpin RNA approach was utilized to knock down the CPT1A gene expression. We employed transwell and lymphatic vessel formation assay to examine invasion and lymphangiogenesis of HDLEC (Human dermal lymphatic endothelial cells). RT-qPCR and westernblot analyses were used to determine genes expression in HDLEC and breast cancer cells. Finally, we determined the relative rate of acetyl-CoA/CoA in shNC and shCPT1A HDLEC cells by LC-MS approach. RESULTS: Knockdown of CPT1A in breast cancer cells (MCF-7 and MDA-MB-231) abolished invasion and lymphangiogenesis of HDLEC cells. Mechanistically, CPT1A depletion suppressed the expression of VEGF-C and VEGF-D in MCF-7 and MDA-MB-231 cells. Interestingly, CPT1A knockdown in HDLEC cells exhibited attenuated expression of lymphangiogenic markers (podoplanin, VEGFR-3, VEGF-C, VEGF-D and PROX-1). Consistently, CPT1A -null HDLEC cells displayed compromised invasion and lymphangiogenesis compared with negative control. Further investigation revealed that CPT1A regulated VEGFR3 via acetyl-CoA mediated H3K9ac, which could be abrogated by supplement of acetate. CONCLUSIONS: In present study, we revealed the mechanism by which CPT1A regulates breast cancer-associated invasion and lymphangiogenesis. Our findings provide insights into CPT1A -promoted breast tumor metastasis and provide rationale for understanding breast cancer metastasis.

Chronic Obstructive Pulmonary Disease as a Risk Factor for Cognitive Dysfunction: A Meta-Analysis of Current Studies.

Cognitive dysfunction has been shown to be associated with many risk factors, such as smoking, diabetes, and body mass index. Chronic obstructive pulmonary disease (COPD), a common disease within the elderly population, has also been found to be related to cognitive decline. However, whether COPD is a risk factor of cognitive dysfunction is not well established. The purpose of this meta-analysis is to investigate the role COPD plays in cognitive dysfunction. PubMed, Cochrane library and Web of Science databases were searched. Three cohort studies and eleven cross-sectional studies were found to be eligible. According to our results, COPD patients had a higher risk of cognitive dysfunction than controls (OR [odds ratio]: 1.72; 95% CI, 1.12-2.65; p = 0.01). The exacerbation of COPD was strongly correlated with cognitive decline. COPD patients performed worse than controls on the Mini- Mental State Examination test, but the results were not statistically significant (OR: -0.79; 95% CI, [-1.78, 0.19]; p = 0.11). Thus, more attention should be given to the occurrence of cognitive decline in COPD patients. The prevention and control of COPD exacerbation are critical.

[Preparation of monoclonal antibodies against flagellin core protein of Vibrio parahaemolyticus and its activity analysis].

OBJECTIVE: To prepare monoclonal antibodies (mAbs) against flagellin core protein of Vibrio (V.) parahaemolyticus and establish the double-sandwich ELISA for testing V.parahaemolyticus from food products. METHODS: BALB/c mice were immunized with flagellin which was extracted from V.parahaemolyticus ATCC17802 through differential centrifugation. The splenocytes from the immunized mice were fused with Sp2/0 myeloma cells in log-phase growth when the antibody titer in serum was 1:32 000. The hybridoma cell lines were screened by regular subcloning approach and used to generate ascites. And mAbs reacting to V.parahaemolyticus flagellin were obtained by purifying from the ascites. RESULTS: Six hybridoma cell lines secreting mAbs against V.parahaemolyticus flagellin were prepared and named VpNo.1-VpNo.6. The mAb titer in serum by indirect ELISA was 1:2×10(6);. The mAbs were subjected to Ig classes and subclasses detection and Western blotting. The Ig subclasses of these mAbs were IgG1, IgG1, IgM, IgG1, IgG2a and IgM, respectively. SDS-PAGE showed that the flagellin protein molecular weight (Mr;) was 42 000. Using VpNo.6, Double-sandwich ELISA kit was set up and applied in detecting V.parahaemolyticus, and its sensitivity was 10(3); CFU/mL. The ELISA showed VpNo.6 had a desirable specificity to V.parahaemolyticus in testing 134 V.parahaemolyticus and 74 non-V.parahaemolyticus strains. The lowest limit of detection was 21 CFU/g sample in the base-material addition test. CONCLUSION: The monoclonal antibodies against flagellin core protein of V.parahaemolyticus were successfully prepared. The double-sandwich ELISA we established using VpNo.6 mAb had a sensitivity as high as 10(3); CFU/mL. The monoclonal antibody of VpNo.6 was highly specific to V.parahaemolyticus.

XAF1 contributes to dengue virus-induced apoptosis in vascular endothelial cells.

The mechanism of vascular leakage in severe dengue infection remains unclear. Here, we used primary human umbilical vein endothelial cells (HUVECs) and the EA.hy926 cell line to study the molecular events that occur after dengue virus serotype 2 (DENV2) infection. DENV2-induced apoptosis was confirmed using nuclear staining, TUNEL assay, and electron microscopy. A genome-wide transcriptome analysis was performed using a microarray of DENV2-infected HUVECs. Notably, interferon-inducible genes were differentially expressed after DENV2 infection. Prominent among these genes was the X chromosome-linked inhibitor of apoptosis protein (XIAP)-associated factor 1 (XAF1; up-regulated 1.2-fold in the microarray analysis and ∼8-fold by qRT-PCR after DENV2 infection). XAF1 protein levels were up-regulated after DENV2 infection in both HUVECs and EA.hy926 cells. Evidence indicated interaction between XAF1 and XIAP during DENV2 infection based on their cellular localization, as observed by confocal microscopy and the coimmunoprecipitation of XIAP with an anti-XAF1 antibody. Next, recombinant EA.hy926 cell lines in which XAF1 was either knocked down or overexpressed were constructed. The expression levels of the apoptosis-related genes caspase 3, caspase 8, caspase 9, and poly-(ADP-ribose) polymerase (PARP) were down-regulated in the XAF1 knockdown (24-48 h postinfection) but were up-regulated in XAF1 overexpressing cells (36 h postinfection). This is the first study of the role of XAF1 in promoting apoptosis in vascular endothelial cells after DENV2 infection.

Formononetin protects neurons against hypoxia-induced cytotoxicity through upregulation of ADAM10 and sAßPPα.

Formononetin, an active constituent of the Chinese herb Astragali Radix, has been reported to have beneficial effects for Alzheimer's disease (AD). Yet the mechanism of this effect remains to be elucidated. The present study shows that formononetin increases soluble-AßPPα (sAßPPα) secretion and thus protects human-AßPP Swedish mutation cell (N2a-AßPP cell) from hypoxia-induced apoptosis. Using hypoxic N2a-AßPP cell as an in vitro model of AD-like pathology, we confirmed that regular treatment with formononetin could have neuroprotective effects, followed respectively by reduced caspase 3 activity and increased cell viability. Strikingly, our data revealed that the caspase 3-blocking effect of formononetin was largely mediated by stimulation of α-secretase cleavage of AßPP, and increasing the secretion of its soluble form, sAßPPα. Moreover, the protective effect of formononetin was totally inhibited by TAPI-2, an α-secretase complex inhibitor, suggesting the role of the sAßPPα pathway in the neuroprotective response to formononetin. We also found that the stimulative effect of formononetin on α-secretase activity was mainly conducted by upregulating ADAM10 expression at the transcriptional level. Altogether, our study provides novel insights into how formononetin mediates stimulation of the ADAM10-sAßPPα pathway and exerts a neuronal protective effect.