Analysis of Macrophage Chemotactic Activity and TLR9 Signaling Pathway in the Mouse Model of Viral Acute Lung Injury.

This study focused on the chemotactic activity of macrophages and the role of the TLR9 signaling pathway in the pathogenesis of viral Acute Lung Injury (ALI). For this purpose, a total of 40 male SPF mice were used, aged 5-8 weeks. They were randomly divided into an experimental group and a control group. The experimental group was further divided into S1 and S2, and the control group was further divided into D1 and D2, with 10 in each. The different groups were detected for the expression of inflammatory cytokines and chemokines and the expression of alveolar macrophages. Results showed that as for the weight, survival status, arterial blood gas analysis, lung index and wet-to-dry value of lung tissue, and lung histopathological analysis results, the S2 group showed more obvious changes versus the D2 group, and the difference was statistically significant (P<0.05). S2 had higher levels of the inflammatory factors TNF-α, IL-1ß, IL-6 and the chemokine CCL3 in the BALF supernatant versus the D2 Group, and the difference is statistically significant (P<0.05). S2 had higher expression levels of chemokines CCR5, TLR9, and JMJD1A mRNA versus the D2 group, and the difference was statistically significant (P<0.05). In conclusion,  the establishment of a mouse ALI model induced by poly l:C was successful; AM has a certain chemotactic activity on CCL3; polyI:C can promote the expression activity and chemotactic activity of macrophages CCR5 through signal pathways, such as TLR9.

Upregulation of Excision Repair Cross-Complementation Group 6-Like (ERCC6L) Promotes Tumor Growth in Hepatocellular Carcinoma.

BACKGROUND: Excision repair cross-complementation group 6-like (ERCC6L) is overexpressed in some malignancies; however, its role in hepatocellular carcinoma (HCC) remains to be further investigated. AIMS: In the present study, we explored the expression and function of ERCC6L in HCC. METHODS AND RESULTS: We investigated the expression of ERCC6L by microarray analysis, using the Cancer Genome Atlas database, and by HCC tissue microarray. The results showed that ERCC6L expression was upregulated in tumor specimens and HCC cell lines. High ERCC6L expression in tumor tissues was significantly correlated with poor prognosis and could serve as an independent prognostic indicator for HCC patients. Results of in vitro and in vivo assays revealed that ERCC6L substantially promoted cell proliferation, and our flow cytometry analysis revealed that this was accomplished by acceleration of the G1/S transition. Finally, gene set enrichment analysis and western blotting results indicated that ERCC6L might regulate HCC proliferation by activating p53 signaling. CONCLUSIONS: Our study suggests that ERCC6L plays an important role in HCC proliferation and that it might serve as a promising therapeutic target in HCC.

Preferences heterogeneity of health care utilization of community residents in China: a stated preference discrete choice experiment.

BACKGROUND: To tackle the issue with the low usage of primary healthcare service in China, it is essential to align resource distribution with the preferences of the community residents. There are few academic researches for describing residents' perceived characteristics of healthcare services in China. This study aims to investigate the preferences of healthcare services utilization in community residents and explore the heterogeneity. The findings will be useful for the policy makers to take targeted measures to tailor the provision of healthcare services. METHODS: The face-to-face interviews and surveys were conducted to elicit four key attributes (care provider; mode of services; cost; travel time) of the preference from community residents for healthcare utilization. A rational test was presented first to confirm the consistency, and then 16 pairs of choice tasks with 12 sociodemographic items were given to the respondents. Two hypothetical options for each set, without an opt-out option, were presented in each choice task. The latent class analysis (LCA) was used to analyse the data. RESULTS: Two thousand one hundred sixty respondents from 36 communities in 6 cities were recruited for our study. 2019 (93.47%) respondents completed valid discrete choice experiment (DCE) questionnaires. The LCA results suggested that four groups of similar preferences were identified. The first group (27.29%) labelled as "Comprehensive consideration" had an even preference of all four attributes. The second group (37.79%) labelled as "Price-driven" preferred low-price healthcare services. The third group labelled as "Near distance" showed a clear preference for seeking healthcare services nearby. The fourth group (34.18%) labelled as "Quality seeker" preferred the healthcare service provided by experts. Willingness to pay (WTP) results showed that people were willing to accept CNY202.12($29.37) for Traditional Chinese Medicine (TCM) services and willing to pay CNY604.31($87.81) for the service provided by experts. CONCLUSIONS: Our study qualitatively measures the distinct preferences for healthcare utilization in community residents in China. The results suggest that the care provider, mode of services, travel time and cost should be considered in priority setting decisions. The study, however, reveals substantial disagreement in opinion of TCM between different population subgroups.

Factors influencing the utilization of postpartum visits among rural women in China.

Postpartum visits (PPVs) are still underutilized in rural China, and identification of factors that influence PPV use is important in ensuring the utilization of maternal health services and for wellbeing of women. A cross-section study was undertaken to collect related data from 347 rural women interviewed six weeks or more after delivery, and an ANOVA was performed to find whether there were significant differences in the number of PPVs among different rural areas in China. According to Andersen's socio-behavioral model of health service use, factors were divided into equitable and inequitable ones. Chi-squared test, univariate and multiple analyses were used to determine the equity of PPV use by identifying factors that were most strongly associated with the use of a PPV. The results showed that 20.2% of the respondents (n=70) did not receive any PPVs, and 62.5% (n=173) of those who had PPVs (n=277) did not receive standard PPVs (referring to at least 3 visits). There was no significant difference among different rural areas in terms of the number of PPVs (F=1.514, P=0.211). Multiple regression analyses revealed that enabling factors such as compensation for delivery expense [OR (95% CI)=2.825 (1.331, 5.995)], village type [OR (95% CI)=1.802 (1.021, 3.182)] and service quality [OR (95% CI)=1.847 (1.074, 3.176)] were strongly associated with PPV use. Both enabling factors such as home visits [OR (95% CI)=1.855 (1.085, 3.174)], service quality [OR (95% CI)=1.993 (1.155, 3.439)] and need factors such as low birth weight [OR (95% CI)=4.424 (1.482, 13.203)] were significantly associated with standard PPV use. Our results suggested that the equitable access to PPVs has been considerably improved in rural areas in China. The associations between inequitable factors and PPV use warrant further exploration, and policies aimed at improving quality and patterns of service supply are needed in order to ensure a full equitable access to maternal health services.

Responses of vegetation cover to hydro-climatic variations in Bosten Lake Watershed, NW China.

Amidst the backdrop of global climate change, it is imperative to comprehend the intricate connections among surface water, vegetation, and climatic shifts within watersheds, especially in fragile, arid ecosystems. However, these relationships across various timescales remain unclear. We employed the Ensemble Empirical Mode Decomposition (EEMD) method to analyze the multifaceted dynamics of surface water and vegetation in the Bosten Lake Watershed across multiple temporal scales. This analysis has shed light on how these elements interact with climate change, revealing significant insights. From March to October, approximately 14.9-16.8% of the areas with permanent water were susceptible to receding and drying up. Both the annual and monthly values of Bosten Lake's level and area exhibited a trend of initial decline followed by an increase, reaching their lowest point in 2013 (1,045.0 m and 906.6 km2, respectively). Approximately 7.7% of vegetated areas showed a significant increase in the Normalized Difference Vegetation Index (NDVI). NDVI volatility was observed in 23.4% of vegetated areas, primarily concentrated in the southern part of the study area and near Lake Bosten. Regarding the annual components (6 < T < 24 months), temperature, 3-month cumulative NDVI, and 3-month-leading precipitation exhibited the strongest correlation with changes in water level and surface area. For the interannual components (T≥ 24 months), NDVI, 3-month cumulative precipitation, and 3-month-leading temperature displayed the most robust correlation with alterations in water level and surface area. In both components, NDVI had a negative impact on Bosten Lake's water level and surface area, while temperature and precipitation exerted positive effects. Through comparative analysis, this study reveals the importance of temporal periodicity in developing adaptive strategies for achieving Sustainable Development Goals in dryland watersheds. This study introduces a robust methodology for dissecting trends within scale components of lake level and surface area and links these trends to climate variations and NDVI changes across different temporal scales. The inherent correlations uncovered in this research can serve as valuable guidance for future investigations into surface water dynamics in arid regions.

Upregulation of Nei-Like DNA Glycosylase 3 Predicts Poor Prognosis in Hepatocellular Carcinoma.

BACKGROUND: Accumulating evidence has suggested that Nei-like DNA glycosylase 3 (NEIL3) is associated with human tumors. However, there are few studies on the role of NEIL3 in hepatocellular carcinoma (HCC). The aim of this study was to investigate the expression profile of NEIL3 and its clinical relevance in HCC. MATERIALS AND METHODS: A total of 130 HCC and corresponding nontumor tissues were collected to perform immunohistochemistry (IHC). The clinical relevance and prognostic value of NEIL3 in HCC were analyzed by the chi-square test, Kaplan-Meier analysis, the Cox proportional hazard model, and nomogram. RESULTS: IHC showed that the NEIL3 protein level was remarkably upregulated in tumor tissues compared with nontumor tissues (fold change = 1.24; P < 0.001). High NEIL3 expression was significantly correlated with BCLC stage (P=0.004) and TNM stage (P=0.005). Overall survival (OS) and disease-free survival (DFS) rates in the high NEIL3 expression group were significantly worse than those in the low NEIL3 expression group (P=0.007 and P=0.004, respectively). Furthermore, subgroup analysis showed that high NEIL3 expression predicted worse OS and DFS for HCC patients with advanced TNM stage, poorly differentiated tumor, HBsAg positive, or cirrhosis. Multivariate analysis and the prognostic nomograms revealed that tumor NEIL3 level may serve as a promising prognostic indicator for OS and DFS in HCC patients. CONCLUSION: Our findings suggested that NEIL3 might be a potential prognosis assessment marker and therapeutic target for HCC patients.

UBE2M promotes cell proliferation via the ß-catenin/cyclin D1 signaling in hepatocellular carcinoma.

Upregulated ubiquitin-conjugating enzyme E2M (UBE2M) is associated with poor prognosis in malignancies; However, the phenotype and mechanism of action of UBE2M in hepatocellular carcinoma (HCC) remain elusive. Here, we report that UBE2M is overexpressed and correlated with poor prognosis in HCC patients. The UBE2M level is an independent prognostic factor for HCC patients. UBE2M knockdown inhibits HCC cell proliferation, migration, and invasion, whereas its overexpression has an opposite effect. Mechanistically, upregulated UBE2M exerts oncogenic effects by translocation of accumulated ß-catenin from the cytoplasm to the nucleus, thus activating downstream ß-catenin/cyclin D1 signaling. In summary, our study demonstrates a notable role of UBE2M in promoting the growth of HCC, providing a novel strategy for HCC prevention and treatment.

[Effects of adenoviral-mediated mda-7/IL-24 gene infection on the growth and drug-resistance of drug-resistant].

OBJECTIVE: To study the effects of tumor suppressor gene mda-7/IL-24 on the growth and drug-resistance of two drug-resistant ovarian cancer cell lines OVCAR-3, OVCAR-8/TR. METHODS: Adenoviral-mediated mda-7/IL-24 (Ad. mda-7/IL-24) was constructed in our previous study, and then applied to infect two cell lines. Western blot analysis was used to detect the expression of MDA-7/IL-24 protein. The cell viability and resistance against four chemotherapeutic drugs, including DDP, ADM, 5-FU and Taxol, were examined with methyl thiazolyl tetrazolium (MTT) rapid photocolorimetric assay. RESULTS: All OVCAR-8/TR and OVCAR-3 cells were infected under the condition that Ad. mda-7/IL-24 was 10 microL, and successfully infected cells could express the MDA-7/IL-24 protein. OVCAR-3 and OVCAR-8/TR cell growth were significantly inhibited after cells infected by Ad. mda-7/IL-24. The cell viability decreased to 10. 5% and 35. 8% respectively in the fifth day. The sensitivity of infected OVCAR-3 or OVCAR-8/TR to Taxol was increased up to 2. 76 or 1. 52 times, as compared with the control. The infected OVCAR-3 cells also had higher sensitivity to ADM and 5-FU, which digitally increased respectively 1. 6 and 3. 01 times when compared to the sensitivities before cells infected. CONCLUSION: The mda-7/IL-24 gene shows cancer growth inhibition on drug-resistance ovarian caner cell lines. Ad. mda-7/IL-24 infecting cell may have the capability of reversing the resistance of tumor cell against Taxol.

[Ad. mda-7/IL-24 construction and expression in infected drug resistant cell line of human ovarian cancer].

OBJECTIVE: To construct the recombinant Ad. mda-7/IL-24 using Adeasy 1 system. METHODS: The mda-7/IL-24 DNA sequence was, by PCR, amplified from the plasmid pREP4-mda-7 and sub-cloned into the shuttle vector pAdTrack CMV. The resultant plasmid and pAdEasy 1 were used to co-transfer into the E. coli BJ5183 cells to undergo homologous recombination. The linearized recombinant plasmid DNA was transferred into 293 cells. Then the ovarian cancer drug resistant cell line OVCAR-8/TR was infected by the recombinant adenovirus, in which the expression of MDA-7/IL-24 protein was detected by Western-blot analysis. RESULTS: The recombinant Ad. mda-7/IL-24 was constructed successfully and approved by sequence analysis, electrophoresis and expression of MDA-7/IL-24 protein. All OVCAR-8/TR cells had been infected under concentration of Ad. mda-7/IL-24 was 10 microL. OVCAR-8/TR cell had the objective protein expression after infected. CONCLUSION: The recombinant adenovirus Ad. mda-7/IL-24 is successfully constructed, which lays a foundation for studying mda-7/IL-24 gene in the field of ovarian cancer drug resistant.

Heat shock protein 70 downregulation inhibits proliferation, migration and tumorigenicity in hepatocellular carcinoma cells.

The overexpression of heat shock protein 70 (HSP70), a major stress-inducible heat shock protein, has been identified to enhance the proliferation, survival, invasion and metastasis of diverse types of human cancer. However, its role in hepatocellular carcinoma (HCC) remains poorly understood. The present study demonstrated that HSP70 expression was higher in tested HCC cell lines, compared with the normal hepatocyte LO2, and the suppression of HSP70 significantly inhibited the proliferation of SMMC-7721 and Hep3B cells. The growth inhibitory effect was mediated by cell cycle arrest at the G1/S phase with reduced cyclin D1 and increased p27Kip1 expression. Furthermore, HSP70 knockdown significantly inhibited the migration and invasion abilities of HCC cells. In conclusion, HSP70 is a key regulator involved in the proliferation, migration and invasion of HCC, and it may be used as a potential therapeutic target for HCC.

Effect of saracatinib on pulmonary metastases from hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. Src is involved in multiple processes of cancer metastasis; however, its significance in HCC is not well defined. In the present study, overexpression of Src phosphorylation (Y416) was observed in the highly metastatic MHCC97H cell line; additionally, through inhibition of Src kinase activation, HCC cell proliferation, migration, invasion and colony formation were significantly reduced in vitro. Tumour growth was not affected in the orthotopic xenograft HCC model, but the metastasic potential was inhibited as revealed by reduced lung metastasic foci after administration of saracatinib. Phosphorylation level of Src pathway signalling molecules, such as Src, FAK and Stat3, were also reduced in vitro and in vivo, as a result of the anti-metastasic effects caused by saracatinib treatment. In conclusion, we demonstrated the pro-metastasic role of Src in HCC, and further experiments suggest the use of the Src inhibitor in combination with cytotoxic agents and other anticancer treatments to improve HCC prognosis.

Prognostic significance of eukaryotic initiation factor 4E in hepatocellular carcinoma.

PURPOSE: Aberrant expression of eukaryotic initiation factor 4E (eIF4E) has been observed in human malignancies. However, its role in hepatocellular carcinoma (HCC) remains to be established. The purpose of this study was to detect eIF4E expression and to evaluate its clinical relevance. METHODS: The eIF4E expression was studied in ninety HCC and randomly selected thirty-one non-tumor tissues from the same patient cohort, as well as in normal hepatic and HCC cell lines. The relation between its expression and clinicopathological parameters was also analyzed. RESULTS: eIF4E expression was higher in HCC samples and cell lines compared with that in non-tumor tissues (P < 0.001) and hepatocyte LO2, respectively. eIF4E overexpression was significantly associated with tumor number (P = 0.005) and incomplete encapsulation (P = 0.001). The 5-year overall survival rate and disease-free survival rate for patients with high eIF4E expression were 32.5 and 31.2 %, respectively; and for low eIF4E expression, it was 67.9 and 64.4 %, respectively (P < 0.001). Furthermore, subgroup analysis showed that high eIF4E level predicted poorer overall survival only for incomplete encapsulation (P = 0.001) and cirrhosis (P < 0.001), but not for complete encapsulation (P = 0.804) and non-cirrhosis (P = 0.359). Multivariate analysis revealed that eIF4E overexpression was an independent indicator for both overall survival (hazard ratio, 2.015; P = 0.043) and disease-free survival (hazard ratio, 2.666; P = 0.006). CONCLUSIONS: eIF4E protein might result in the malignant progression of HCC, and its overexpression may be a powerful prognostic biomarker and therapeutic target for HCC patients.

Human umbilical mesenchymal stem cells-seeded bladder acellular matrix grafts for reconstruction of bladder defects in a canine model.

BACKGROUND: The goal of this study was to explore the feasibility of utilizing human umbilical mesenchymal stem cells (HUMSCs)-seeded Bladder acellular matrix graft (BAMG) for bladder reconstruction in a canine model. METHODOLOGY/PRINCIPAL FINDINGS: HUMSCs were isolated from newborn umbilical cords and identified by flow cytometry. Partial cystectomy was performed in the experimental and control group. Bladder defects were repaired with HUMSCs-BAMG in the experimental group and repaired with unseeded-BAMG in control group. The implanted grafts were harvested after surgery. H&E and immunohistochemistry staining were performed to evaluate the regeneration of the bladder defect. Primary cultured HUMSCs displayed typical fibroblast morphology with spindle-shaped. Flow cytometry indicated that these cells were positive for CD105 (97.3%) and CD44 (99%), but negative for CD34 (2.8%), CD31 (2.1%), and CD45 (1.7%). Immunohistochemistry staining showed that a multilayered urothelium and well-developed smooth muscle were observed at 12 weeks in experiment group. In contrast, multilayered urothelial tissues were also observed at 12 weeks in group B, but well-developed smooth muscle bundles were observed. CONCLUSIONS/SIGNIFICANCE: Our preliminary results demonstrate that UMSC-seeded BAMGs are superior to unseeded BAMGs to promote the regeneration of bladder defects. Our findings indicated that HUMSCs may be a potential cell source for bladder tissue engineering.

P53 and PCNA is positively correlated with HPV infection in laryngeal epitheliopapillomatous lesions in patiets with different ethnic backgrounds in Xinjiang.

OBJECTIVE: To explore the correlation of human papillomavious (HPV) infection with expression of p53 and proliferating cell nuclear antigen (PCNA) in patients with different ethnicity in Xinjiang, China. METHODS: 166 biopsy specimens from 83 laryngeal squamous cell carcinomas (LSCC), 63 laryngeal papillomas (LP), and 20 laryngeal inflammatory polyps (LIP) were included in this study. HPV infection was determined by polymerase chain reaction (PCR) using specific types of HPV primers. Expression of p53 and PCNA was assessed using immunohistostaining. RESULTS: The frequency of HPV 6/11 was higher in LP (33.3%) than in LSCC (9.6%) (P<0.0005), whereas the frequency of HPV 16/18 was higher in LSCC (37.3%) than in LP (6.3%) (P<0.0005). Patients of the Han ethnic group with LSCC had a higher infection rate with HPV 6/11 or HPV 6/11 and HPV 16/18 coinfection than those of Uygur and Kazak ethnicity (P<0.05). Overexpression of p53 and PCNA were higher in LSCC (62.7%, 57.8%) than in LP (38%, 33.3%) (P<0.005, and P<0.005, respectively). That of p53 was not associated with lymph-node metastases and clinical stages, but overexpression of PCNA closely correlated with clinical stage. CONCLUSIONS: These results strongly implicate HPV6/11 infection in the carcinogenesis of LSCC and LP, respectively. There was a higher coincidence of increased malignancy of laryngeal tumors with overexpression of p53 and PCNA. Overexpression of p53 may serve as an early risk marker for malignant transformation in HPV infected cells while the overexpression of PCNA may serve as a late marker for progression of LSCC.

Altered distribution and expression pattern of E-cadherin in hepatocellular carcinomas: correlations with prognosis and clinical features.

OBJECTIVE: E-cadherin has been identified as a tumor suppressor in many types of carcinoma. However, some studies recently suggested that the role and expression of E-cadherin might be more complex and diverse. In the present study, we evaluated the prognostic value of E-cadherin expression with reference to levels in membranes and cytoplasm, and the membrane/cytoplasm ratio, in hepatocellular carcinomas (HCCs) after curative hepatectomy. METHODS: The expression of E-cadherin was assessed by immunohistochemistry in HCC tissue microarrays from 125 patients, and its prognostic values and other clinicopathlogical data were retrospectively analyzed. Patients were followed for a median period of 43.7 months (range 1 to 126 months). RESULTS: Univariate analysis demonstrated that a high membrane/cytoplasm (M/C) ratio of E-cadherin expression was associated with poor overall survival (OS) (P =0.001) and shorter time to recurrence (TTR) (P =0.038), as well as tumor size, intrahepatic metastasis, and TNM stage. In contrast, neither membrane nor cytoplasmic expression of E-cadherin was related with OS and TTR. Furthermore, multivariate analysis confirmed the M/C ratio to be an independent predictor of OS (P =0.031). ?2 tests additionally showed that the M/C ratio of E-cadherin expression was related with early stage recurrence (P =0.012), rather than later stage recurrence. CONCLUSION: The M/C ratio of E-cadherin expression is a strong predictor of postoperative survival and is associated with early stage recurrence in patients with HCC.

[Effect of adenovirus-mediated mda-7/IL-24 gene infection on apoptosis of drug-resistant human ovarian cancer cell lines OVCAR-3 and OVCAR-8/TR].

BACKGROUND & OBJECTIVE: Melanoma differentiation associated gene-7 (mda-7/IL-24) has double functions: specifically induces tumor cell apoptosis and modulates immune responses. Therefore, it is a strong candidate for human cancer gene therapy. This study was to evaluate the effect of adenovirus-mediated mda-7/IL-24 infection on the apoptosis of drug-resistant ovarian cancer cell lines OVCAR-3 and OVCAR-8/TR. METHODS: Adenovirus-mediated mda-7/IL-24 (Ad.mda-7/IL-24) was constructed using AdEasy 1 system. OVCAR-3 and OVCAR-8/TR cells were infected by Ad.mda-7/IL-24. The expression of MDA-7/IL-24 protein was detected by Western blot. Cell apoptosis was detected by flow cytometry with Hoechst33258 staining. Cell cycle distribution was detected by flow cytometry. RESULTS: The recombinant Ad.mda-7/IL-24 was confirmed by DNA sequencing and electrophoresis. The expression of MDA-7/IL-24 protein was detected in the cells after infection. Within 72 h after Ad.mda-7/IL-24 infection, the maximal apoptosis rates of OVCAR-3 and OVCAR-8/TR cells were 14.1% and 32.4%, respectively, significantly higher than empty vector group and uninfected group. CONCLUSIONS: The recombinant Ad.mda-7/IL-24 was successfully constructed. It can induce apoptosis in drug-resistant ovarian cancer OVCAR-3 and OVCAR-8/TR cells.