PI3K/AKT signaling pathway associates with pyroptosis and inflammation in patients with endometriosis.

Endometriosis (EMS) is known to be closely associated with inflammation. We evaluate the possible mechanism linking the PI3K/AKT signaling pathway with pyroptosis and inflammation in EMS. We collected 30 patients undergoing laparoscopic for endometriosis as the EMS group and those undergoing surgery for uterine fibroids as the control group, from whom we collected serum, normal endometrium, eutopic endometrium and ectopic endometrium. Transmission electron microscopy (TEM) was used to observe the internal structure of endometrial cells. Western Blot was used to detect the protein expression of PI3K, P-PI3K, AKT, P-AKT, NLRP3, Caspase-1, GSDMD, and GSDMD-N. Immunohistochemistry (IHC) staining was used to detect the expression of PI3K, AKT, NLRP3, Caspase-1, GSDMD, and GSDMD-N proteins. Immunofluorescence (IF) staining was used to observe the expression of GSDMD-N. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze the mRNA levels of PI3K, AKT, NLRP3, Caspase-1, GSDMD, and GSDMD-N. ELISA was used to detect serum levels of IL-1ß, IL-18, TLR4, and NF-κB. We found that activation of PI3K/AKT signaling pathway in endometriosis significantly increased the level of cellular pyroptosis and inflammatory factors. Our results suggest that there is a positive correlation between the PI3K/AKT signaling pathway and pyroptosisas well as inflammation in EMS patients.

Fine root dynamics and its contribution to soil organic carbon stocks with Caragana intermedia plantation development in alpine sandy land.

Shrubs are the main species in desert ecosystems. Better understanding shrubs fine root dynamics and its contribution to soil organic carbon (SOC) stocks can improve the accuracy of carbon sequestration estimation and provide basic data for the calculation of carbon sequestration potential. The ingrowth core method was used to investigate the fine root (< 1 mm in diameter) dynamics of Caragana intermedia Kuang et H. C. Fu plantation with different age (4-, 6-, 11-, 17-, and 31-year-old) in Gonghe Basin of the Tibetan Plateau, and the annual fine root mortality was used for calculation the annual carbon input to SOC pool. The results showed that fine root biomass, production, and mortality first increased and then decreased as the plantation age increased. Fine root biomass peaked in 17-year-old plantation, production and mortality peaked in 6-year-old plantation, and turnover rate of 4- and 6-year-old plantations were significantly higher than other plantations. Fine root production and mortality were negative correlated with soil nutrients at depth of 0-20 and 20-40 cm. The variation range of carbon input by fine root mortality across different plantation age at 0-60 cm soil depth was 0.54-0.85 Mg ha-1 year-1, accounting for 2.40-7.54% of the SOC stocks. C. intermedia plantation has a strong carbon sequestration potential from long time scale. Fine roots regenerate faster in young stands and lower soil nutrients environment. Our results suggest that the influences of plantation age and soil depth should be taken into account when calculating the contribution of fine root to SOC stocks in desert ecosystems.

Bushen Wenyang Huayu Decoction inhibits autophagy by regulating the SIRT1-FoXO-1 pathway in endometriosis rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Bushen Wenyang Huayu Decoction (BWHD) is a traditional Chinese medicine for tonifying kidney and warming Yang, thereby resolving blood stasis and relieving pain. BWHD can significantly improve the clinical symptoms of patients with endometriosis (EMs), but its mechanism is still unclear. AIM OF THE STUDY: We evaluated the expression and role of the SIRT1-FoxO-1 pathway and autophagy levels in EMs rats. The therapeutic effects and potential therapeutic mechanisms of BWHD were also investigated. METHODS: Twenty rats were randomized into the sham group and eighty rats were used for model establishment by autologous transplantation. After successful modeling, they were randomized into the model, BWHD, EX527+BWHD and EX527 groups, with 20 rats in each group. All rats were intragastrically administered with for 3 weeks. Localization of Sirtuin 1 (SIRT1), Forkhead boxO-1 (FoXO-1), Beclin-1, autophagy-related 5 (Atg5) and autophagy-related 7 (Atg7) was determined by immunohistochemical staining. The expression of the above proteins was determined by Western blot and their messenger RNA (mRNA) levels were detected by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). RESULTS: The protein and mRNA expressions of FoXO-1, Beclin-1, Atg5 and Atg7 in the model group were markedly increased, while that of SIRT1 was markedly decreased relative to the sham group (p < 0.05 and p<0.01, respectively). Results showed that the protein and mRNA expressions of FoXO-1, Beclin-1, Atg5 and Atg7 in eutopic and ectopic endometrium of BWHD group were lower, while SIRT1 expression was higher than in the model group (p < 0.05 and p<0.01, respectively). Furthermore, protein and mRNA expression levels of FoXO-1, Beclin-1, Atg5 and Atg7 in eutopic and ectopic endometrium of EX527 group were higher, while SIRT1 level was significantly lower than in the model group (p < 0.05 and p < 0.01, respectively). The EX527-induced changes in protein and mRNA expressions were reversed in the EX527+BWHD group (p < 0.05 and p < 0.01, respectively). CONCLUSIONS: BWHD inhibits autophagy by up-regulating SIRT1 and down-regulating FoXO-1 expression in EMs via the SIRT1-FoXO-1 signaling pathway. Therefore, it is a potential treatment for EMs.

Bushen Wenyang Huayu Decoction Targets TLR4/NF-κB Mediated Autophagy to Treat Endometriosis Effectively.

Endometriosis has been found to be closely related to autophagy. This study aimed to elucidate the possible mechanism of Bushen Wenyang Huayu Decoction (BWHD) in treating endometriosis (EMs) by targeting TLR4/NF-κB-mediated autophagy. Autologous grafting was used to generate the EMs model in rats. Once the model was developed, BWHD high-dose and low-dose groups received intragastric administration of BWHD, and the gestrinone group served as a positive control. Immunofluorescence labeling and Western blotting were used for the protein expression of toll-like receptor 4 (TLR4), nuclear transcription factor-κB (NF-κB), Beclin-1, and selective autophagy connector protein P62 (P62). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze mRNA levels of TLR4, NF-κB, Beclin-1, and P62. We found that BWHD significantly reduced the size of ectopic lesions in rats with EMs, regulated reproductive hormone levels, and alleviated the cell autophagy level. It suggested that BWHD could be an effective treatment of EMs by targeting TLR4/NF-κB signaling pathway.

Involvement of bradykinin and bradykinin B1 receptor in patients with endometriosis.

Endometriosis (EM), a benign aseptic inflammatory disease, is associated with the presence of endometrial foci. Pain, one of its typical symptoms, has been reported as a constant stressor, but the etiology and pathogenesis of EM-associated pain are unclear. In the present study, eutopic and ectopic endometrium samples from women with EM (n=50) and normal endometrium samples from control subjects (n=20) were collected. Serum levels of prostaglandin E2 (PGE2), prostaglandin F2α (PGF2α) and bradykinin (BK) were measured using commercial ELISA kits. The expression of the BKB1 receptor (BKB1R) protein was evaluated by immunohistochemical staining and western blot assay. The mRNA expression of BKB1R was measured by reverse transcription-quantitative PCR. The results revealed that there was a substantial increase in the protein and mRNA expression of BKB1R, as well as the release of PGE2, PGF2α and BK in the blood, in the EM group compared with that in the control group. Moreover, PGE2, PGF2α and BK levels were significantly correlated with each other, as well as with the pain intensity of EM. The increased expression levels of BKB1R protein and mRNA were positively correlated with the pain degree of EM. Thus, these data indicated that BK and BKB1R were involved in the pathological onset of EM-associated pain and that they may play an important role in EM-related pain by inducing PGE2 and PGF2α. The data indicate a potential new therapeutic target for EM-related pain.

Nectin-1 Is an Entry Mediator for Varicella-Zoster Virus Infection of Human Neurons.

Varicella-zoster virus (VZV) maintains lifelong latency in neurons following initial infection and can subsequently be reactivated to result in herpes zoster or severe neurological manifestations such as encephalitis. Mechanisms of VZV neuropathogenesis have been challenging to study due to the strict human tropism of the virus. Although neuronal entry mediators of other herpesviruses, including herpes simplex virus, have been identified, little is known regarding how VZV enters neurons. Here, we utilize a human stem cell-based neuronal model to characterize cellular factors that mediate entry. Through transcriptional profiling of infected cells, we identify the cell adhesion molecule nectin-1 as a candidate mediator of VZV entry. Nectin-1 is highly expressed in the cell bodies and axons of neurons. Either knockdown of endogenous nectin-1 or incubation with soluble forms of nectin-1 produced in mammalian cells results in a marked decrease in infectivity of neurons. Notably, while addition of soluble nectin-1 during viral infection inhibits infectivity, addition after infection has no effect on infectivity. Ectopic expression of human nectin-1 in a cell line resistant to productive VZV infection confers susceptibility to infection. In summary, we have identified nectin-1 as a neuronal entry mediator of VZV. IMPORTANCE Varicella-zoster virus (VZV) causes chickenpox, gains access to neurons during primary infection where it resides lifelong, and can later be reactivated. Reactivation is associated with shingles and postherpetic neuralgia, as well as with severe neurologic complications, including vasculitis and encephalitis. Although the varicella vaccine substantially decreases morbidity and mortality associated with primary infection, the vaccine cannot prevent the development of neuronal latency, and vaccinated populations are still at risk for reactivation. Furthermore, immunocompromised individuals are at higher risk for VZV reactivation and associated complications. Little is known regarding how VZV enters neurons. Here, we identify nectin-1 as an entry mediator of VZV in human neurons. Identification of nectin-1 as a neuronal VZV entry mediator could lead to improved treatments and preventative measures to reduce VZV related morbidity and mortality.

[Effects of precipitation on water use characteristics of Caragana intermedia plantations with different stand ages in alpine sandy land]. / 降雨对高寒沙地不同林龄中间锦鸡儿水分利用特征的影响.

To clarify the changes of water sources for Caragana intermedia plantations at different ages (4, 9, 17 and 31 years) in response to rainfall in the Gonghe Basin of Qinghai Province, China, we used the stable isotope technique to identify δ2H and δ18O compositions of soil water, xylem water, groundwater, and rain water before and after rainfalls. The proportions of different water sources were calculated by the Iso-Source model. The results showed that the δ2H and δ18O compositions of the shallow soil layer (0-40 cm) of all plantations responded significantly to the precipitation. The isotopic values of plant xylem water, soil water, and groundwater of each plantation were spotted on the lower right of the local meteoric water line (LMWL) either before or after rainfall, with lower intercepts and slopes than LMWL and the global meteoric water line (GMWL). The isotopic compositions of xylem water and soil water of C. intermedia plantations were closer to LMWL after rainfall. The 4- and 9-year-old C. intermedia plantations mainly used shallow soil water, the 17-year-old plantation mainly used middle layer soil water (40-90 cm), and the 31-year-old plantation primarily use deep soil water before rainfall. After rainfall, the shallow soil layer became sources of water absorption for all plantations. The utilization proportions of groundwater for all plantations were only 1.8%-11.9%. In general, water sources of different aged C. intermedia plantations showed similar responses to rainfall, by primarily absorbing the shallow soil water supplied by rainfall and reducing the use of groundwater.

Fine root dynamic characteristics and effect on plantation's carbon sequestration of three Salix shrub plantations in Tibetan Plateau alpine sandy land.

Desertification land in Gonghe Basin of Tibetan Plateau, China accounts for 91.9% of the total land area. Vegetation restoration and reconstruction with desert shrubs in degraded ecosystem are effective ways to prevent and control desertification. However, the evaluation studies of fine root dynamic characteristics of desert shrubs and their contribution to carbon sequestration of plantation are limited. To gain a better understanding of vegetation restoration, the vertical distribution of fine root biomass, fine root decomposition, fine root turnover was investigated, as well as their coupling effect on carbon sequestration of plantation in three desert vegetation. The results estimated that the total decomposition time of fine roots of Salix cheilophila (S. cheilophila), Salix psammophila (S. psammophila), and Salix microstachya (S. microstachya) are 39.00, 27.99 and 35.95 years. Biomass carbon density for three Salix plantations ranged from 1.42 to 2.39 t/hm2, which showed that three Salix plantations in alpine sandy land are an important carbon pool. In addition, fine root biomass carbon density for the three shrub plantations varied significantly. Fine root biomass carbon density for S. psammophila reached the largest among the three plantations, which was 1.48 t/hm2, accounting for the ratio of 62% of the plantation total biomass carbon density. The results indicated that the root system of S. psammophila, especially the fine roots, was very developed, which was conducive to soil water transportation and carbon sequestration. Therefore, S. psammophila might be a better species for carbon sequestration of plantation in alpine sandy areas. The carbon input from the fine roots of the three shrub plantations through decomposition and turnover into the plantations accounts for 11.5% to 15.5% of total carbon sequestration of plantations. Therefore, the fine roots dynamics must be considered for long-term carbon pool estimations in three Salix plantations, otherwise the total carbon sequestration of plantations would be underestimated.

FOXD3­AS1/miR­128­3p/LIMK1 axis regulates cervical cancer progression.

Long non­coding RNA forkhead box D3 antisense RNA 1 (FOXD3­AS1) functions as an oncogenic regulator in several types of cancer, including breast cancer, glioma and cervical cancer. However, the effects and mechanisms underlying FOXD3­AS1 in cervical cancer (CC) are not completely understood. The present study aimed to investigate the biological functions and potential molecular mechanisms underlying FOXD3­AS1 in CC progression. Reverse transcription­quantitative PCR was performed to detect FOXD3­AS1, microRNA (miR)­128­3p and LIM domain kinase 1 (LIMK1) expression levels in CC tissues and cells. Immunohistochemical staining and western blotting were conducted to assess LIMK1 protein expression levels in CC tissues and cells, respectively. Cell Counting Kit­8 and BrdU assays were used to determine the role of FOXD3­AS1 in regulating cell proliferation. CC cell migration and invasion were assessed by performing Transwell assays. Dual­luciferase reporter assays were conducted to verify the binding between miR­128­3p and FOXD3­AS1. FOXD3­AS1 expression was significantly increased in CC tissues and cell lines compared with adjacent healthy tissues and normal cervical epithelial cells, respectively. High FOXD3­AS1 expression was significantly associated with poor differentiation of tumor tissues, increased tumor size and positive lymph node metastasis. FOXD3­AS1 overexpression significantly increased CC cell proliferation, migration and invasion compared with the negative control (NC) group, whereas FOXD3­AS1 knockdown resulted in the opposite effects compared with the small interfering RNA­NC group. Moreover, the results demonstrated that FOXD3­AS1 targeted and negatively regulated miR­128­3p, which indirectly upregulated LIMK1 expression. Therefore, the present study demonstrated that FOXD3­AS1 upregulated LIMK1 expression via competitively sponging miR­128­3p in CC cells, promoting CC progression.

Occurrence, source and ecotoxicological risk assessment of pesticides in surface water of Wujin District (northwest of Taihu Lake), China.

This study investigated the occurrence and distribution of pesticides in surface water (lakes, major rivers and tributaries) and potential discharge sources (fish ponds, livestock and poultry farms, and sewage treatment plants) in Wujin District (northwest of Taihu Lake), Jiangsu province, China. An analytical liquid chromatography-tandem mass spectrometry method was developed for 38 pesticides, which was applied in the monitoring of 240 surface water samples and 76 potential discharge source samples. Eleven insecticides and five fungicides with temporal and spatial variation were detected in surface water. The total pesticide concentrations in surface water in different seasons were as follows: March > August > June > November. The two most polluting and widespread pesticides were carbendazim (maximum concentration 508 ng L-1, detection rate 100%) and imidacloprid (maximum concentration 438 ng L-1, detection rate 88%). Gehu Lake (S46) and Sanshangang River (S12) were seriously polluted water bodies. Seven insecticides and four fungicides were detected in the potential discharge sources; and their composition changed significantly with the seasons. The concentrations of detected organophosphorus pesticides and neonicotinoids (e.g. acetamiprid in March and dichlorvos in November) in a few non-agricultural planting sources were far greater than those detected in surface water, and hence a few fish ponds, livestock and poultry farms, and sewage treatment plants might be the potential discharge sources of pesticides in the surrounding surface water. The estimated input flux of the studied pesticides from upstream rivers to Taihu Lake was 141.95 kg a-1. Furthermore, more attention should be paid to the medium or high aquatic ecotoxicological risk presented by the levels of organophosphorus pesticides, carbamates, and benzimidazoles.

Patients With Natural Killer (NK) Cell Chronic Active Epstein-Barr Virus Have Immature NK Cells and Hyperactivation of PI3K/Akt/mTOR and STAT1 Pathways.

BACKGROUND: Chronic active Epstein-Barr virus (CAEBV) presents with high levels of viral genomes in blood and tissue infiltration with Epstein-Barr virus (EBV)-positive lymphocytes. The pathogenesis of CAEBV is poorly understood. METHODS: We evaluated 2 patients with natural killer (NK) cell CAEBV and studied their NK cell phenotype and signaling pathways in cells. RESULTS: Both patients had increased numbers of NK cells, EBV predominantly in NK cells, and immature NK cells in the blood. Both patients had increased phosphorylation of Akt, S6, and STAT1 in NK cells, and increased total STAT1. Treatment of 1 patient with sirolimus reduced phosphorylation of S6 in T and B cells, but not in NK cells and did not reduce levels of NK cells or EBV DNA in the blood. Treatment of both patients' cells with JAK inhibitors in vitro reduced phosphorylated STAT1 to normal. Patients with T- or B-cell CAEBV had increased phosphorylation of Akt and S6 in NK cells, but no increase in total STAT1. CONCLUSIONS: The increase in phosphorylated Akt, S6, and STAT1, as well as immature NK cells describe a new phenotype for NK cell CAEBV. The reduction of STAT1 phosphorylation in their NK cells with JAK inhibitors suggests a novel approach to therapy.

Correction: Spatial Heterogeneity of Soil Nutrients after the Establishment of Caragana intermedia Plantation on Sand Dunes in Alpine Sandy Land of the Tibet Plateau.

[This corrects the article DOI: 10.1371/journal.pone.0124456.].

Metformin-induced autophagy and irisin improves INS-1 cell function and survival in high-glucose environment via AMPK/SIRT1/PGC-1α signal pathway.

In order to explore the protective function of metformin on pancreatic ß cells to alleviate insulin resistance and underlying mechanisms, INS-1 cells were cultured into normal control (N), high glucose (H), high glucose and metformin (H + Met), high glucose and chloroquine (H + CQ), and high glucose and Ex527 (H + Ex527) groups, respectively. Upon 24-hr cultivation, the proliferation and glucose-stimulated insulin secretion (GSIS) of INS-1 cells were determined, and the expression of irisin and other proteins associated with AMPK/SIRT1/PGC-1α signal pathway, autophagy, and apoptosis was evaluated. Compared with the N group, the cells from the H group revealed lower proliferation, GSIS, and expression of irisin and proteins associated with AMPK/SIRT1/PGC-1α signal pathway and autophagy, but higher expression of proteins associated with apoptosis; in contrast, metformin could significantly rescue lower cell proliferation, GSIS, and expression of proteins associated with AMPK/SIRT1/PGC-1α signal pathway and autophagy, as well as irisin, and suppress apoptosis in high-glucose environment. Meanwhile, autophagy inhibitor CQ and SIRT1 inhibitor Ex527 can block above functions of metformin. Therefore, metformin can promote INS-1 cell proliferation, enhance GSIS, and suppress apoptosis by activating AMPK/SIRT1/PGC-1α signal pathway, up-regulating irisin expression, and inducing autophagy in INS-1 cells in high-glucose environment.

Fine-root decomposition characteristics of four typical shrubs in sandy areas of an arid and semiarid alpine region in western China.

BACKGROUND AND AIMS: Research into the variability of fine-root decomposition and nutrient cycling processes in arid and semiarid ecosystems is highly significant not only for investigations of regional and global carbon and nitrogen cycling but also for offering a theoretical basis for vegetation restoration and reconstruction. In particular, information is limited on fine-root decomposition processes and nutrient releasing characteristics in the high-altitude Qinghai Gonghe basin, which has different tree species and variable fine-root diameters. MATERIALS AND METHODS: Four types of Salicaceae and Caragana shrubs were selected at the Qinghai Gonghe desert ecosystem research station. The litterbag method was adopted to measure decomposition rates of fine-roots with three diameter classes (1-2 mm, 0.5-1 mm, and 0-0.5 mm). Chemical analysis was performed to determine nutrient (C, N, P, and K) concentrations of fine-root, and nutrient release rates were compared among fine-roots with different diameters during different decomposition periods. The differences in mass residual ratio and nutrient release rate among different diameter classes were studied with one-way ANOVA. RESULTS: Fine-root decomposition rates were in the order Caragana intermedia > Caragana korshinskii > Salix psammophila > Salix cheilophila. Fine-root decomposition showed a trend of "fast-slow-fast" variation, and decomposition rate increased as the diameter of fine-roots increased, irrespective of tree species. During the decomposition process, the nutrients C, N, and P of fine-root were in a release state for the four shrubs with different fine-root diameters, and the corresponding release rates of Caragana shrubs were higher than those of Salicaceae shrubs. Release rates of nutrients C and N accelerated as fine-root diameter increased, whereas release rates of nutrients P and K had no observed relation with fine-root diameter. Fine-root decomposition ratio was significantly correlated with initial values of N, P, C/N, C/P, and N/P of fine-root. Fine-root mass loss ratio was significantly correlated with initial concentration of soil nutrient K, and the correlation was positive for fine-roots with diameters of 0-0.5 mm and 0.5-1 mm; however, no other significant correlation was observed between fine-root mass loss ratio and initial soil environmental factors within this study. CONCLUSIONS: Our study showed that tree species and fine-root diameter strongly affected decomposition rates, whereas diameter class exerted little effect on nutrient release rates.

Epstein-Barr Virus and the Human Leukocyte Antigen Complex.

PURPOSE: While most adults are infected Epstein-Barr virus (EBV), 3-5% remain uninfected. The human leukocyte antigen (HLA) complex, which controls many pathogens, may influence infection and disease associated with EBV. RECENT FINDINGS: Numerous EBV proteins and miRNAs down-regulate HLA class I and II expression on the cell surface. HLA class II functions as a receptor for EBV entry into B cells. Specific HLA class II alleles correlate with the susceptibility of B cells to EBV infection in vitro and with EBV seropositivity or seronegativity of humans. HLA class I polymorphisms correlate with development and severity of EBV infectious mononucleosis and with the risk of several virus-associated malignancies including nasopharyngeal carcinoma, Hodgkin lymphoma, and post-transplant lymphoproliferative disease. SIGNIFICANCE: These findings indicate that while EBV has evolved to use MHC class II as a receptor for virus entry, polymorphisms in MHC class II and class I influence virus infection and disease.

Dynamics of biomass and carbon sequestration across a chronosequence of Caragana intermedia plantations on alpine sandy land.

Considering the variations in carbon concentrations among different plant components can significantly improve carbon storage estimates. The aim of this study was to estimate the biomass and carbon storage and sequestration in Caragana intermedia Kuang et H. C. Fu plantations for six different ages. The biomass and carbon sequestration in shrub biomass were quantified using a destructive method that involved analysing the carbon concentrations in the leaves, branches, stem bark, stem wood, roots > 5 mm, 2 mm < roots ≤ 5 mm, and roots ≤ 2 mm. The biomass and biomass carbon density of the C. intermedia plantations increased with the age of the stands. The biomass carbon density of the woody components was at its maximum in the 30-year-old plantation (14.27 ± 2.71 t·ha-1), indicating that C. intermedia plantations in alpine sandy land are an important carbon pool. The carbon accumulation rate of the woody components was higher during the early stages of plantation development. The carbon concentrations differed significantly among the components but changed only slightly with the stand age. The weighted mean carbon concentration of the woody components, which was found to be 44%, must be considered when estimating the long-term carbon pools in C. intermedia plantations.

Ultrarapid Measurement of Diagnostic Antibodies by Magnetic Capture of Immune Complexes.

Rapid point-of-care, antibody-based testing is not currently available for the diagnosis of most autoimmune and infectious diseases. Here we report a simple, robust and ultrafast fluid-phase immunocapture method for clinical measurements of antibody levels. This method employs neodymium magnetic sticks that capture protein A/G-coated paramagnetic beads bound to antibody-luciferase-labeled antigen complexes. We demonstrate the ability to effectively measure specific antibody levels in serum samples from patients with varied infectious or autoimmune disorders, and in the case of Sjögren's syndrome directly in saliva, requiring about a minute per assay. We also show the feasibility of coupling this method with a hand-held luminometer for portable testing. Our method offers the potential to quickly diagnose a multitude of autoimmune and infectious diseases in point-of-care settings.

HLA-DQ ß1 alleles associated with Epstein-Barr virus (EBV) infectivity and EBV gp42 binding to cells.

Epstein-Barr virus (EBV) infects B cells and ~95% of adults are infected. EBV glycoprotein gp42 is essential for entry of virus into B cells. EBV gp42 binds to the ß1 chain of HLA-DQ, -DR, and -DP on B cells, and uses these molecules for infection. To investigate if certain HLA-DQ alleles are associated with EBV seronegativity, we recruited ~3,300 healthy adult blood donors, identified 106 EBV-seronegative individuals, and randomly selected a control group of EBV-seropositive donors from the donor pool. A larger than expected proportion of EBV-seronegative subjects were HLA-DQ ß1 *04/*05 and *06/*06, and to a lesser extent, *02/*03, compared with the control group, while a larger than expected portion of EBV-seropositive persons were HLA-DQ ß1 *02/*02. We examined the ability of EBV gp42 to bind to different HLA-DQ molecules using human and mouse cells stably expressing these alleles. EBV gp42 bound less effectively to cells expressing HLA-DQ ß1 *04/*05, *06/*06, or *03/*03 than to cells expressing HLA-DQ ß1 *02/*02. These data are consistent with our observations of increased EBV seronegativity with DQ ß1 *04/*05 or *06/*06 alleles. These findings emphasize the importance of a single genetic locus (HLA-DQ ß1) to influence infectivity with EBV.

The dynamic monitoring of aeolian desertification land distribution and its response to climate change in northern China.

Aeolian desertification is poorly understood despite its importance for indicating environment change. Here we exploit Gaofen-1(GF-1) and Moderate Resolution Imaging Spectroradiometer (MODIS) data to develop a quick and efficient method for large scale aeolian desertification dynamic monitoring in northern China. This method, which is based on Normalized Difference Desertification Index (NDDI) calculated by band1 &band2 of MODIS reflectance data (MODIS09A1). Then we analyze spatial-temporal change of aeolian desertification area and detect its possible influencing factors, such as precipitation, temperature, wind speed and population by Convergent Cross Mapping (CCM) model. It suggests that aeolian desertification area with population indicates feedback (bi-directional causality) between the two variables (P < 0.05), but forcing of aeolian desertification area by population is weak. Meanwhile, we find aeolian desertification area is significantly affected by temperature, as expected. However, there is no obvious forcing for the aeolian desertification area and precipitation. Aeolian desertification area with wind speed indicates feedback (bi-directional causality) between the two variables with significant signal (P < 0.01). We infer that aeolian desertification is greatly affected by natural factors compared with anthropogenic factors. For the desertification in China, we are greatly convinced that desertification prevention is better than control.

Cell Surface THY-1 Contributes to Human Cytomegalovirus Entry via a Macropinocytosis-Like Process.

Previously we showed that THY-1 has a critical role in the initial stage of infection of certain cell types with human cytomegalovirus (HCMV) and that THY-1 is important for HCMV-mediated activation of phosphatidylinositol 3-kinase (PI3K)/Akt during virus entry. THY-1 is known to interact with integrins and is a major cargo protein of clathrin-independent endocytic vesicles. Since macropinocytosis involves integrin signaling, is PI3K/Akt dependent, and is a clathrin-independent endocytic process, we determined whether THY-1 has a role in HCMV entry by macropinocytosis. Using electron microscopy in two cell lines that support HCMV infection in a THY-1-dependent manner, we found that HCMV enters these cells by a macropinocytosis-like process. THY-1 associated with HCMV virions on the cell surface and colocalized with virus inside macropinosomes. 5-(N-Ethyl-N-isopropyl)amiloride (EIPA) and soluble THY-1 blocked HCMV infection in the cell lines by ≥80% and 60%, respectively. HCMV entry into the cells triggered increased influx of extracellular fluid, a marker of macropinocytosis, and this increased fluid uptake was inhibited by EIPA and by soluble THY-1. Blocking actin depolymerization, Na+/H+ exchange, PI3K, and Pak1 kinase, which are critical for macropinocytosis, impaired HCMV infection. Neither internalized HCMV virions nor THY-1 in virus-infected cells colocalized with transferrin as determined by confocal microscopy, indicating that clathrin-mediated endocytosis was not involved in THY-1-associated virus entry. These results suggest that HCMV has adapted to utilize THY-1, a cargo protein of clathrin-independent endocytotic vesicles, to facilitate efficient entry into certain cell types by a macropinocytosis-like process. IMPORTANCE: Human cytomegalovirus (HCMV) infects over half of the population and is the most common infectious cause of birth defects. The virus is the most important infection occurring in transplant recipients. The mechanism of how HCMV enters cells is controversial. In this study, we show that THY-1, a cell surface protein that is critical for the early stage of entry of HCMV into certain cell types, contributes to virus entry by macropinocytosis. Our findings suggest that HCMV has adapted to utilize THY-1 to facilitate entry of HCMV into macropinosomes in certain cell types. Further knowledge about the mechanism of HCMV entry into cells may facilitate the development of novel inhibitors of virus infection.