Epigenetic dynamics of aging and cancer development: current concepts from studies mapping aging and cancer epigenomes.

PURPOSE OF REVIEW: This review emphasizes the role of epigenetic processes as incidental changes occurring during aging, which, in turn, promote the development of cancer. RECENT FINDINGS: Aging is a complex biological process associated with the progressive deterioration of normal physiological functions, making age a significant risk factor for various disorders, including cancer. The increasing longevity of the population has made cancer a global burden, as the risk of developing most cancers increases with age due to the cumulative effect of exposure to environmental carcinogens and DNA replication errors. The classical 'somatic mutation theory' of cancer cause is being challenged by the observation that multiple normal cells harbor cancer driver mutations without resulting in cancer. In this review, we discuss the role of age-associated epigenetic alterations, including DNA methylation, which occur across all cell types and tissues with advancing age. There is an increasing body of evidence linking these changes with cancer risk and prognosis. SUMMARY: A better understanding about the epigenetic changes acquired during aging is critical for comprehending the mechanisms leading to the age-associated increase in cancer and for developing novel therapeutic strategies for cancer treatment and prevention.

Tissue-location-specific transcription programs drive tumor dependencies in colon cancer.

Cancers of the same tissue-type but in anatomically distinct locations exhibit different molecular dependencies for tumorigenesis. Proximal and distal colon cancers exemplify such characteristics, with BRAFV600E predominantly occurring in proximal colon cancers along with increased DNA methylation phenotype. Using mouse colon organoids, here we show that proximal and distal colon stem cells have distinct transcriptional programs that regulate stemness and differentiation. We identify that the homeobox transcription factor, CDX2, which is silenced by DNA methylation in proximal colon cancers, is a key mediator of the differential transcriptional programs. Cdx2-mediated proximal colon-specific transcriptional program concurrently is tumor suppressive, and Cdx2 loss sufficiently creates permissive state for BRAFV600E-driven transformation. Human proximal colon cancers with CDX2 downregulation showed similar transcriptional program as in mouse proximal organoids with Cdx2 loss. Developmental transcription factors, such as CDX2, are thus critical in maintaining tissue-location specific transcriptional programs that create tissue-type origin specific dependencies for tumor development.

CD206 modulates the role of M2 macrophages in the origin of metastatic tumors.

Tumor metastasis is a key factor affecting the life of patients with malignant tumors. For the past hundred years, scientists have focused on how to kill cancer cells and inhibit their metastasis in vivo, but few breakthroughs have been made. Here we hypothesized a novel mode for cancer metastasis. We show that the phagocytosis of apoptotic tumor cells by macrophages leads to their polarization into the M2 phenotype, and that the expression of stem cell related as well as drug resistance related genes was induced. Therefore, it appears that M2 macrophages have "defected" and have been transformed into the initial "metastatic cancer cells", and thus are the source, at least in part, of the distal tissue tumor metastasis. This assumption is supported by the presence of fused cells with characteristics of both macrophage and tumor cell observed in the peripheral blood and ascites of patients with ovarian cancer. By eliminating the expression of CD206 in M2 macrophages using siRNA, we show that the growth and metastasis of tumors was suppressed using both in vitro cell line and with experimental in vivo mouse models. In summary, we show that M2 macrophages in the blood circulation underwent a "change of loyalty" to become "cancer cells" that transformed into distal tissue metastasis, which could be suppressed by the knockdown of CD206 expression.

Fear of COVID-19 among cancer patients in Henan Province, Central China: causes, results, and coping factors.

Objectives: Cancer patients exhibit fear of COVID-19, which could lead to serious consequences. However, minimal information is available about the effect of the COVID-19 pandemic on the mental health of cancer patients. Therefore, this study aims to examine the fear level of COVID-19 among cancer patients in Henan Province, Central China and to identify its causes, results, and coping factors. Methods: An online survey was conducted among 1,067 cancer patients. The participants reported their individual fear level of COVID-19, risk of COVID-19 infection, risk of death from COVID-19, COVID-19 vaccination concerns, influence level of COVID-19 pandemic on their disease treatment, loneliness due to COVID-19, economic burden from COVID-19, quality of life, safety behavior, information regarding COVID-19 vaccination, psychological guidance, physical activities, and demographic characteristics. Chi-square and cumulative logistic regression were used to determine the predictors of COVID-19 fear level. Results: This study indicates that cancer patients report moderate fear level of COVID-19 in Central China (66.9%). The six cause factors (risk of COVID-19 infection, risk of death from COVID-19, COVID-19 vaccination concerns, influence level of COVID-19 pandemic on disease treatment, loneliness due to COVID-19, and economic burden from COVID-19) were positively associated with COVID-19 fear level. Three coping factors (information regarding COVID-19 vaccination, psychological guidance, and physical activities) were negatively associated with COVID-19 fear level. COVID-19 fear level was negatively associated with quality of life and positively associated with safety behavior. Conclusion: Our results suggest that governments should improve access to personalized vaccine counseling and psychological guidance by undertaking the responsibility of patients' attending physicians and increasing publicity. Physical activities should be included in the treatment program to help cancer patients better recover their physical and mental health.

Prognostic value of the geriatric nutritional index in colorectal cancer patients undergoing surgical intervention: A systematic review and meta-analysis.

Background: We reviewed the literature to assess the prognostic ability of the geriatric nutritional risk index (GNRI) for patients with colorectal cancer (CRC) undergoing curative surgery. Methods: The online databases of PubMed, CENTRAL, ScienceDirect, Embase, and Google Scholar were searched for articles reporting the relationship between GNRI and outcomes in CRC patients. English language studies were searched up to 28th April 2022. Results: Ten studies with 3802 patients were included. Meta-analysis indicated that patients with low GNRI had significantly poor overall survival (HR: 2.41 95% CI: 1.72, 3.41 I2 = 68%) and disease-free survival (HR: 1.92 95% CI: 1.47, 2.49 I2 = 49%) as compared to those with high GNRI. The meta-analysis also indicated a significantly higher risk of complications with low GNRI as compared to high GNRI (HR: 1.98 95% CI: 1.40, 2.82 I2 = 0%). The results did not change on subgroup analysis based on study location, age group, GNRI cut-off, and sample size. Conclusion: Current evidence indicates that GNRI can be a valuable prognostic indicator for CRC patients undergoing surgical intervention. Patients with low GNRI have poor overall and disease-free survival and a higher incidence of complications. Clinicians could use this simple indicator to stratify patients and formulate personalized treatment plans. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/, identifier (CRD42022328374).

Putative Prevention of XML Injection Against Myocardial Ischemia Is Mediated by PKC and PLA2 Proteins.

Background: Xinmailong (XML) injection is a CFDA-approved traditional Chinese medicine with clinical value for heart failure treatment. The present investigation was aimed to evaluate the potential protective roles of this injection on myocardial ischemia and the underlying molecular mechanism. Methods: In our study, we selected two models of myocardial ischemia rats. Rats were randomly divided into six groups, with saline or XML administrated 4 days before ischemia model establishment. ECG of different time intervals and biochemical parameters of end point were measured. The potential mechanisms of the protective role of XML were explored using system pharmacology and molecular biology approaches. Results: Myocardial ischemia rats demonstrated abnormal ECG and serum levels of cTnT. Pretreatment with XML significantly attenuated these damages, especially the medium doses. GO and KEGG analysis revealed that the 90 putative target genes were associated with pathways of fatty acid absorption/metabolism, inflammation, RAAS, and vascular smooth muscle. Further network pharmacology method identified five main chemical ingredients and potential targets of XML injection for myocardial ischemia. Mechanically, the beneficial effect of XML injection was mediated by the reactive oxygen species (ROS) inhibition and inflammation attenuation via regulating the expression levels of targets of PKC and PLA2. Conclusion: These findings indicate that XML exerts protective effects against myocardial injury, with attenuated ROS production, apoptosis, and inflammation. Therefore, we speculate that XML may be an alternative supplementary therapeutic agent for myocardial ischemia prevention.

Mechanism exploration of ancient pharmaceutic processing (Paozhi) improving the gastroprotective efficacy of Aucklandiae Radix.

ETHNOPHARMACOLOGICAL RELEVANCE: Processing, also called Paozhi in Chinese, is an ancient Chinese pharmaceutic processing technique developed along with the Chinese herbal medicines (CHMs). The understanding of the mechanism of Paozhi has been investigated for several decades. Aucklandiae Radix (CAR) and its roasted processed products are all used in indigestion as a kind of CHMs. Processed Aucklandiae Radix (PAR) had a stronger effect to protect gastric mucosa than CAR, while the main compounds in CAR were reduced sharply after being processed. The underlying mechanism of this phenomenon is still unclear. AIM OF THE STUDY: This study was aimed to evaluate whether PAR have a stronger gastroprotective effect than CAR and the underlying mechanisms of such circumstance. MATERIALS AND METHODS: Ultra-fast liquid chromatography coupled with quadrupole time of flight mass spectrometry (UFLC-QTOF-MS/MS) coupled with multivariate statistical analyses was employed to explore chemical compounds which had a relatively stable content in PAR. Based on the compounds selected as the research object, network pharmacology was applied to visualize the relationships between the selected components and the gastroprotective-related targets from disease database, at the same time the possible intervention path of CAR/PAR which might be responsible for the effect of CAR/PAR on gastritis-induced rats was also built. Then, the key proteins were detected by western blotting to verify and compare the pharmacological effects of CAR/PAR. RESULTS: Through UFLC-QTOF-MS/MS and orthogonal partial least squares discriminant analysis (OPLS-DA), sixteen compounds stable in PAR were discovered, of which saussureamine C and saussureamine B were estimated as the core compounds to exert gastroprotective in PAR predicted by network pharmacology analysis. Under the guide of KEGG pathway enrichment analysis, PI3K/AKT, p38 MAPK (Mitogen-activated protein kinase) and nuclear factor-kappa B (NF-κB) signaling pathways were forecasted as the possible healing mechanisms of CAR/PAR, and that result was verified by the experiments in vivo. PAR performed a stronger ability to reduce the level of p38 MAPK and NF-κB p65 than CAR, which may partially explain the different ability of CAR/PAR against gastric mucosa damage. CONCLUSION: This study clarified that although Paozhi entailed a sharp decrease on the main compounds of CAR, there were some compounds which were not sensitive to high temperature and preserved in PAR and had a relative higher content in PAR than in CAR. PAR has stronger influence on MAPKs/NF-κB signaling pathway than CAR, which may reveal that the stronger gastroprotective effect of PAR perhaps rely on the constitutions with a higher relative abundance after Paozhi. The present research combined UFLC-QTOF-MS/MS and network pharmacology deeply investigated the impact of the roasted processing on the chemical constitutions and gastroprotective effect of CAR and offered reference for the clinical application of CAR/PAR.

CIDO-COVID-19: An Ontology for COVID-19 Based on CIDO.

To realize integration, organization and reusability of knowledge related to COVID-19, an ontology for COVID-19 (CIDO-COVID-19) was constructed which extended the Coronavirus Infectious Disease Ontology (CIDO) by adding terms of COVID-19 related to symptoms, prevention, drugs and clinical domains. First, terms from the existing ontologies, literature, clinical guidelines and other resources about COVID-19 were merged. Then, the Stanford seven-step approach was used to define and organize the acquired terms. Finally, the CIDO-COVID-19 was built on basis of the terms mentioned above using Protégé. The CIDO-COVID-19 is a more comprehensive ontology for COVID-19, covering multiple areas in the domain of COVID-19, including disease, diagnosis, etiology, virus, transmission, symptom, treatment, drug and prevention.Clinical Relevance- The CIDO-COVID-19 covers multiple areas related to COVID-19, including diseases, diagnosis, etiology, virus, transmission, symptoms, treatment, drugs, prevention. Compared with the CIDO, it is expanded to cover drugs, prevention, and clinical domain. The definition of terms in CIDO-COVID-19 refers to biomedical ontologies, Clinical glossaries and clinical guidelines for COVID-19, which can provide clinicians with standard terminology in the clinical domain.

Application effect of the principle of seamless management in nursing risk control of the department of gastrointestinal surgery.

OBJECTIVE: To determine the application effect of nursing risk control based on the principle of seamless management in the department of gastrointestinal surgery. METHODS: A total of 62 patients with gastrointestinal diseases admitted to the Department of Gastrointestinal Surgery in our hospital for nursing risk management based on the principle of seamless management were enrolled into a research group, and another 58 patients admitted before the implementation of such management were enrolled into a control group. The two groups were compared in the incidence of complications, Chinese Perceived Stress Scale (CPSS) score, self-rating anxiety scale (SAS) score, nursing satisfaction, and awareness rate of health knowledge, and the nursing quality scores and medical complaints and disputes were compared before and after the implementation of nursing risk control based on the principle of seamless management by 19 nurses in the department. RESULTS: The pulmonary infection rate, incision infection rate, incidence of gastrointestinal reaction, and incidence of anastomotic leakage in the research group were all significantly lower than those in the control group (all P<0.05), and CPSS and SAS scores of the research group were also both significantly lower than those of the control group (both P<0.001). In addition, the research group showed significantly higher nursing satisfaction and awareness rate of health knowledge than the control group (both P<0.01), and there were notably less medical complaints and disputes after implementation of the principle of seamless management (P<0.01). Moreover, after implementation of the principle, nursing staff acquired significantly higher scores of professional skills (health education, professional knowledge, and practical operation; all P<0.001), and also contributed to significantly higher scores of nursing work (basic nursing, disinfection and isolation, ward management, intensive care, document writing, and nursing safety; all P<0.001). CONCLUSION: Nursing risk control based on the principle of seamless management can improve the comprehensive quality of nursing staff in the department of gastrointestinal surgery and the overall quality of nursing, thus lowering the incidence of nursing risk, relieving patients' negative moods, and improving the nurse-patient relationship and satisfaction of both nurses and patients.

Could the connectedness of primary health care workers involved in social networks affect their job burnout? A cross-sectional study in six counties, Central China.

BACKGROUND: This study aimed to reveal the effects of the connectedness of primary health care (PHC) workers in social networks on their job burnout. METHODS: Cross-sectional survey data of rural PHC workers in China were analyzed. A total of 663 respondents were enrolled. Chi-square and cumulative logistic regression were used to determine the effects of the connectedness of PHC workers in social networks on their job burnout. RESULTS: PHC workers in rural China had high levels of emotional exhaustion (24.1%), depersonalization (15.7%), and lack of personal accomplishment (34.7%). More than half of the participants were in the middle connectedness level in terms of their advisory (70.4%) and friendship (70.3%) networks. The degree of emotional exhaustion seemed to increase when participants had a low connectedness in their friendship networks (ß = 0.769, 95% CI = 0.080-1.458, P = 0.029). Respondents with the middle level of connectedness in advisory networks had higher levels of depersonalization (ß = 0.739, 95% CI = 0.130-1.348, P = 0.017) and lack of personal accomplishment (ß = 0.583, 95% CI = 0.111-1.055, P = 0.015) than those with the high degree of connectedness in advisory networks. CONCLUSIONS: The connectedness of PHC workers in social networks influenced their job burnout. Thus, organizations should establish an informal communication platform and information feedback mechanism, promote and manage friendship networks, and help PHC workers overcome emotional exhaustion. Managers should also encourage individuals with a high level of connectedness in advisory networks play the role of "opinion leader" so that they can help others mitigate burnout.

Overexpression of DUSP6 enhances chemotherapy-resistance of ovarian epithelial cancer by regulating the ERK signaling pathway.

Objective: DUSP6 is a negative regulator of the ERK signaling pathway and plays an important role in chemotherapy-resistance. Previously we showed that DUSP6 is overexpressed in ovarian cancer side population (SP) cells that possess cancer stem cell-like properties and are quiescent and chemotherapy-resistant. Here, we explore the effects of DUSP6 on chemotherapy-resistance by examining its regulation of the ERK signaling pathway and G0/G1 cell cycle arrest. Methods: mRNA and protein expression of DUSP6 and G0/G1 cell cycle checkpoint regulating proteins (CyclinD1, CyclinD3 and CyclinE2) was evaluated among ovarian cancer cell lines and tissue samples. Ovarian cancer cells were transiently transfected to overexpress DUSP6. After treatment with cisplatin, cell viability was measured by the MTS assay at 48 hours and the half maximal inhibitory concentration (IC50) for each cell line was calculated. Subcellular localization and cell cycle analysis were determined by using immunofluorescence and FACS, respectively. Results: SKOV3 and OVCAR8 SP cells were shown to express higher levels of DUSP6 and lower levels of CyclinD3 compared with non-SP (NSP) cells (P<0.001). Among 39 ovarian cancer tissue samples, expression of DUSP6 in the chemotherapy-resistant group (12 samples) was higher than in the chemotherapy-sensitive group (27 samples) (P<0.05). While a lower level of expression of CyclinD3 was seen in the chemotherapy-resistant group, it was not statistically different from the chemotherapy-sensitive group. HO8910 cells where shown to have higher IC50 to cisplatin than SKOV3 or OVCAR8 cells, and this correlated with higher levels of DUSP6 expression. Overexpression of DUSP6 in SKOV3 cells led to an increase in cisplatin IC50 values (P<0.05), and also markedly reduced the expression levels of phospho-ERK1/2 and CyclinD3 and to the predominance of cells in the G0/G1 phase. Conclusion: Our findings reveal an enhancement of chemotherapy-resistance and a predominance of cells in G1 cell cycle arrest in DUSP6-overexpressing ovarian cancer cells. This suggests that overexpression of DUSP6 promotes chemotherapy-resistance through the negative regulation of the ERK signaling pathway, increasing the G0/G1 phase ratio among ovarian cancer cells, and leading to cellular quiescence.

Job burnout and correlated factors of three-tiered public health workers: A cross-sectional study in China.

This study aimed to reveal burnout levels and its potential influencing factors among three-tiered public health workers in China. A total of 1,328 public health workers from cities, counties and townships in Henan and Hubei provinces participated. Cumulative logistic regressions were used to determine job burnout predictors. Township workers (ß = 0.276, p = .046) showed higher levels of emotional exhaustion (EE) than city workers. Municipal workers showed higher levels of depersonalisation (DP) (ß = -0.439, p = .004) and lack of personal accomplishment (LPA) (ß = -0.343, p = .005) than township workers. Compared with those who had better results, township workers with average levels of interpersonal relationships (ß = 0.703, p = .014) exhibited higher EE degrees. Municipal (ß = 1.050, p < .001) and county (ß = 1.359, p < .001) workers with moderate training satisfaction had higher levels of burnout than those with a high degree of training satisfaction. Municipal (ß = 3.933, p < .001) and county (ß = 1.218, p = .018) workers earning 2,000 RMB and below every month were more likely to have higher burnout levels than those earning 4,001 RMB and above. Township respondents exhibited high EE levels, given the low-level education of their services' target audience. Similarly affected by interpersonal relationships, township workers require interpersonal and emotional intervention programmes. City respondents showed a high level of DP and LPA, which was attributed to considerable differences between public health workers and medical workers in cities. Hence, apart from narrowing the reward gap between clinicians and public health workers, improving public health workers' social cognition and status by various channels was found necessary. The burnout of municipal and county respondents were influenced by income and training. Improving the role of income as incentive and reforming trainings programmes would be necessary.

Apoptotic SKOV3 cells stimulate M0 macrophages to differentiate into M2 macrophages and promote the proliferation and migration of ovarian cancer cells by activating the ERK signaling pathway.

Ovarian cancer has a high rate of recurrence, with M2 macrophages having been found to be involved in its progression and metastasis. To examine the relationship between macrophages and ovarian cancer in the present study, M0 macrophages were stimulated with apoptotic SKOV3 cells and it was found that these macrophages promoted tumor proliferation and migration. Subsequently, the mRNAs and proteins expressed at high levels in these M2 macrophages were examined by RNA­Seq and quantitative proteomics, respectively, which revealed that M0 macrophages stimulated by apoptotic SKOV3 cells also expressed M2 markers, including CD206, interleukin­10, C­C motif chemokine ligand 22, aminopeptidase­N, disabled homolog 2, matrix metalloproteinase 1 and 5'­nucleotidase. The abundance of phosphorylated Erk1/2 in these macrophages was increased. The results indicate that apoptotic SKOV3 cells stimulate M0 macrophages to differentiate into M2 macrophages by activating the ERK pathway. These results suggest possible treatments for patients with ovarian cancer who undergo chemotherapy; inhibiting M2 macrophage differentiation during chemotherapy may reduce the rate of tumor recurrence.

Predictor of sleep difficulty among community dwelling older populations in 2 African settings.

Sleep deprivation is a common phenomenon among older population and is commonly linked to behavioral, physiological, and psychosocial factors. Not much is known about sleep deprivation among older population in Africa. Therefore, in this study we aimed to investigate the basic sociodemographic and psychosocial predictors of self-reported sleep deprivation among older population.In this study we analyzed cross-sectional data on 1495 community dwelling men and women aged 50 years and above. Data were collected from the SAGE Well-Being of Older People Study conducted in South Africa and Uganda. Outcome variable was self-reported sleep difficulty last 30 days. Multivariable logistic regression models were used to identify the variables significantly associated with sleep difficulty.The prevalence of mild-moderate sleep difficulty was 32.6% (27.9, 37.6) and severe/extreme 23.0% (20.3, 26.0) respectively. Multivariable analysis revealed that sleep difficulty was associated with several behavioral, environment, and illness conditions. In South Africa, those who reported dissatisfaction with living condition had 1.592 [1.087, 2.787] times higher odds of reporting mild/moderate sleep difficulty. Poor subjective quality of life (QoL) was associated with higher odds of severe/extreme sleep difficulties (odds ratios [OR] = 4.590, 95% confidence interval [CI] = 2.641, 7.977 for South Africa, and OR = 4.461, 95% CI = 2.048 and 9.716 for Uganda). In Uganda, perceived depression was associated with higher odds of severe/extreme (OR = 2.452, 95% CI = 1.073, 5.602) sleep difficulties among men, and both mild/moderate (OR = 1.717; 95% CI = 1.011, 2.914) and severe/extreme sleep difficulties among women (OR = 2.504, 95% CI = 1.408, 4.453).More than half of the participants had sleep difficulty of certain degrees, emphasising an urgent need for intervention for sleep deprivation in the population. Interventions targeting to promote subjective health, quality of life, and living environment may prove beneficial for improving sleep health in this regard.

Stromal vascular fraction promotes migration of fibroblasts and angiogenesis through regulation of extracellular matrix in the skin wound healing process.

BACKGROUND: A refractory wound is a typical complication of diabetes and is a common outcome after surgery. Current approaches have difficulty in improving wound healing. Recently, non-expanded stromal vascular fraction (SVF), which is derived from mature fat, has opened up new directions for the treatment of refractory wound healing. The aim of the current study is to systematically investigate the impact of SVF on wound healing, including the rate and characteristics of wound healing, ability of fibroblasts to migrate, and blood transport reconstruction, with a special emphasis on their precise molecular mechanisms. METHODS: SVF was isolated by digestion, followed by filtration and centrifugation, and then validated by immunocytochemistry, a MTS proliferation assay and multilineage potential analysis. A wound model was generated by creating 6-mm-diameter wounds, which include a full skin defect, on the backs of streptozocin-induced hyperglycemic mice. SVF or human adipose-derived stem cell (hADSC) suspensions were subcutaneously injected, and the wounds were characterized over a 9-day period by photography and measurements. A scratch test was used to determine whether changes in the migratory ability of fibroblasts occurred after co-culture with hADSCs. Angiogenesis was observed with human umbilical vein endothelial cells. mRNA from fibroblasts, endotheliocyte, and skin tissue were sequenced by high-throughput RNAseq, and differentially expressed genes, and pathways, potentially regulated by SVF or hADSCs were bioinformatically analyzed. RESULTS: Our data show that hADSCs have multiple characteristics of MSC. SVF and hADSCs significantly improved wound healing in hyperglycemic mice. hADSCs improve the migratory ability of fibroblasts and capillary structure formation in HUVECs. SVF promotes wound healing by focusing on angiogenesis and matrix remodeling. CONCLUSIONS: Both SVF and hADSCs improve the function of fibroblast and endothelial cells, regulate gene expression, and promote skin healing. Various mechanisms likely are involved, including migration of fibroblasts, tubulogenesis of endothelial cells through regulation of cell adhesion, and cytokine pathways.

Exosomes from Macrophages Exposed to Apoptotic Breast Cancer Cells Promote Breast Cancer Proliferation and Metastasis.

Exosomes have recently become the subject of increasing research interest. Interactions between tumor and host cells via exosomes play crucial roles in the initiation, progression and invasiveness of breast cancer. In our study, we used exosomes isolated from a co-culture model of THP-1-derived macrophages exposed to apoptotic MCF-7 or MDA-MB-231 breast cancer cell line cells to investigate their effects on naïve MCF-7 or MDA-MB-231 cells in vitro and in vivo. This post-chemotherapy tumor microenvironment model allowed us to explore possible mechanisms that explain increased proliferation and metastasis of breast cancer seen in some patients. Our results suggest that while exosomes derived from macrophages normally inhibit proliferation and metastasis of MCF-7 or MDA-MB-231 cells, exposure of macrophages to breast cancer cells that have experienced chemotherapy are modified them to promote these processes. Exosomes from macrophages exposed to apoptotic cancer cells have increased amounts of IL-6 that increases the phosphorylation of STAT3, which likely explains the increased transcription of STAT3 target genes such as CyclinD1, MMP2 and MMP9. These observations suggest that the inhibition of exosome secretion and STAT3 signaling pathway activation might suppress the growth and metastasis of malignant tumors, and provide new targets for therapeutic treatment of malignant tumors after chemotherapy.

Construction of a sensitive pyrogen-testing cell model by site-specific knock-in of multiple genes.

A pyrogen test is crucial for evaluating the safety of drugs and medical equipment, especially those involved in injections. As existing pyrogen tests, including the rabbit pyrogen test, the limulus amoebocyte lysate (LAL) test and the monocyte activation test have limitations, development of new models for pyrogen testing is necessary. Here we develop a sensitive cell model for pyrogen test based on the lipopolysaccharides (LPS) signal pathway. TLR4, MD2, and CD14 play key roles in the LPS-mediated pyrogen reaction. We established a new TLR4/MD2/CD14-specific overexpressing knock-in cell model using the CRISPR/CAS9 technology and homologous recombination to detect LPS. Stimulation of our TLR4/CD14/MD2 knock-in cell line model with LPS leads to the release of the cytokines IL-6 and TNF-alpha, with a detection limit of 0.005 EU/ml, which is greatly lower than the lower limit of 0.015 EU/ml detected by the Tachypleus amebocyte lysate (TAL) assay.

Why Patients with Chronic Disease Keep Silent? Analysis of Item Nonresponse in Rural China.

BACKGROUND: This study aimed to identify the characteristics of item nonresponse and examine the factors affecting the refusal or failure to respond of patients with chronic disease in rural China. METHODS: A cross-sectional survey data from patients with chronic disease from rural China were analyzed. A total of 1,099 patients were enrolled. Chi-square test and cumulative logistic regression determined the predictors of having item nonresponse. RESULTS: The respondents in central provinces (OR = 2.311, 95%CI = 0.532∼1.144, P < 0.001) with over eight household members (OR = 0.067, 95%CI = -1.632∼-0.349, P = 0.002), multiple chronic diseases (OR = 0.301, 95%CI = -1.673∼-0.727, P < 0.001), and low health knowledge level (OR = 2.112, 95%CI = 0.405∼1.090, P < 0.001) had more item nonresponse numbers. Compared with the participants with high school education level and above, the item nonresponse number seemed to increase when the participants were illiterate (OR = 2.159, 95%CI = 0.254∼1.285, P = 0.003), had primary school education (OR = 2.161, 95%CI = 0.249∼1.294, P = 0.004) and junior school education (OR = 2.070, 95%CI = 0.160∼1.296, P = 0.012). CONCLUSION: This study indicates the influencing factors of the item nonresponse in survey of patients with chronic disease in rural China. This study contributes to investigation practice and highlights that health institutions should improve the quality of follow-up services. Moreover, the government should pay more attention to the care of vulnerable groups, especially patients with chronic disease in rural areas.

Application of a TLR overexpression cell model in pyrogen detection.

Pyrogens are components derived from microorganisms that induce complex inflammatory responses. Current approaches to detect pyrogens are complex and difficult to replicate, thus there is a need for new methods to detect pyrogens. We successfully constructed a pyrogen-sensitive cell model by overexpressing Toll-like receptor (TLR)2, TLR4, MD2, and CD14 in HEK293 cells. Since the cytokine IL-6 is specifically released upon stimulation of the TLR2 and TLR4 signaling pathways in response to pyrogen stimulation, we used it as a read out for our assay. Our results show that IL-6 is released in response to trace amounts of pyrogens in our cell model. Pyrogen incubation times and concentrations were explored to determine the sensitivity of our cell model, and was found to be sensitive to 0.05 EU/ml of LPS and 0.05 ug/ml of LTA after stimulation for 5 hr. Our TLR overexpressing cell model, with IL-6 as readout, could be a new method for in vitro testing of pyrogens and applicable for evaluating the safety of drugs.

Analysis of Circulating Tumor Cells in Ovarian Cancer and Their Clinical Value as a Biomarker.

BACKGROUND/AIMS: Monitoring the appearance and progression of tumors are important for improving the survival rate of patients with ovarian cancer. This study aims to examine circulating tumor cells (CTCs) in epithelial ovarian cancer (EOC) patients to evaluate their clinical significance in comparison to the existing biomarker CA125. METHODS: Immuomagnetic bead screening, targeting epithelial antigens on ovarian cancer cells, combined with multiplex reverse transcriptase-polymerase chain reaction (Multiplex RT-PCR) was used to detect CTCs in 211 samples of peripheral blood (5 ml) from 109 EOC patients. CTCs and CA125 were measured in serial from 153 blood and 153 serum samples from 51 patients and correlations with treatment were analyzed. Immunohistochemistry was used to detect the expression of tumor-associated proteins in tumor tissues and compared with gene expression in CTCs from patients. RESULTS: CTCs were detected in 90% (98/109) of newly diagnosed patients. In newly diagnosed patients, the number of CTCs was correlated with stage (p=0.034). Patients with stage IA-IB disease had a CTC positive rate of 93% (13/14), much higher than the CA125 positive rate of only 64% (9/14) for the same patients. The numbers of CTCs changed with treatment, and the expression of EpCAM (p=0.003) and HER2 (p=0.035) in CTCs was correlated with resistance to chemotherapy. Expression of EpCAM in CTCs before treatment was also correlated with overall survival (OS) (p=0.041). CONCLUSION: Detection of CTCs allows early diagnose and expression of EpCAM in CTC positive patients predicts prognosis and should be helpful for monitoring treatment.