Identification and quantification of serum KIN17 protein based on ELISA assay and exploring its clinical diagnostic value in liver cancer.

It has been well-elaborated that KIN17 protein is closely related to the expression, development and prognosis of liver cancer; however, till date, there has been no study about detecting the KIN17 protein in serum, which is important to developing clinical applications. The objective of this work is to detect serum KIN17 protein by the ELISA method and to explore the diagnostic significance of the KIN17 protein in liver cancer. First, we verified the ELISA method for serum KIN17 measurement according to five aspects: accuracy, precision, specificity, stability and detection limit. Results illustrate that the recovery rate of the ELISA method can be controlled between 90% and 110%, the variation coefficient of intra-assay can be controlled within 16%, and the variation coefficient of inter-assay can be controlled within 10%. There is no non-specific reaction with common tumor markers, and the detection limit can reach 0.125 ng mL-1. The results show that the KIN17 protein can be detected by ELISA, and there is a significant rise in KIN17 concentration in a liver cancer group compared with a healthy group, whose average concentrations are 1.730 ng mL-1 and 0.3897 ng mL-1, respectively. On this basis, we hypothesize that the serum KIN17 protein can serve as a potential biomarker of liver cancer and be measurable with the verified ELISA system after specific ultrafiltration and centrifugation, which is of great significance for the diagnosis and treatment of liver cancer.

Prediction of viral pneumonia based on machine learning models analyzing pulmonary inflammation index scores.

OBJECT: To obtain Pulmonary Inflammation Index scores from imaging chest CT and combine it with clinical correlates of viral pneumonia to predict the risk and severity of viral pneumonia using a computer learning model. METHODS: All patients with suspected viral pneumonia on CT examination admitted to The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University from December 2022 to March 2023 were retrospectively selected. The respiratory viruses were monitored by RT-PCR and categorized into patients with viral pneumonia and those with non-viral pneumonia. The extent of lung inflammation was quantified according to the Pulmonary Inflammation Index score (PII). Information on patient demographics, comorbidities, laboratory tests, pathogenetic testing, and radiological data were collected. Five machine learning models containing Random Forest(RF), Radial Basis Function Neural Network (RBFNN), Support Vector Machine (SVM), K Nearest Neighbour Algorithm (KNN), and Kernel Ridge Regression (KRR) were used to predict the risk of onset and severity of viral pneumonia based on the clinically relevant factors or PII. RESULTS: Among the five models, the SVM model performed best in ACC (76.75 %), SN (73.99 %), and F1 (72.42 %) and achieved a better area under the receiver operating characteristic curve (ROC) (0.8409) when predicting the risk of developing viral pneumonia. RF had the best overall classification accuracy in predicting the severity of viral pneumonia, especially in predicting pneumonia with a PII classification of grade I, the RF model achieved an accuracy of 98.89%. CONCLUSION: Machine learning models are valuable in assessing the risk of viral pneumonia. Meanwhile, machine learning models confirm the importance in predicting the severity of viral pneumonia through PII. The establishment of machine learning models for predicting the risk and severity of viral pneumonia promotes the further development of machine learning in the medical field.

Changes of the acute myocardial infarction-related resident deaths in a transitioning region: a real-world study involving 3.17 million people.

Background: The impact of acute myocardial infarction (AMI) on the life span of residents in a transitioning region has not been studied in depth. Therefore, we aimed to evaluate the changes in AMI-related resident deaths in a transitioning region in China. Methods: A longitudinal, population-based study was performed to analyze the deaths with/of AMI in Pudong New Area (PNA), Shanghai from 2005 to 2021. The average annual percentage change (AAPC) of AMI in crude mortality rates (CMR), age-standardized mortality rates worldwide (ASMRW), and rates of years of life lost (YLLr) were calculated by the joinpoint regression. The impact of demographic and non-demographic factors on the mortality of residents who died with/of AMI was quantitatively analyzed by the decomposition method. Results: In 7,353 residents who died with AMI, 91.74% (6,746) of them were died of AMI from 2005 to 2021. In this period, the CMR and ASMRW of residents died with/of AMI were 15.23/105 and 5.17/105 person-years, the AAPC of CMR was 0.01% (95% CI: -0.71,0.72, p = 0.989) and 0.06% (95% CI: -0.71,0.84, p = 0.868), and the ASMRW decreased by 2.83% (95% CI: -3.66,-2.00, p < 0.001) and 2.76% (95% CI: -3.56,-1.95, p < 0.001), respectively. The CMR of people died of AMI showed a downward trend (all p < 0.05) in people ≥60 years but an upward trend [AAPC = 2.47% (95% CI: 0.07,4.94, p = 0.045)] in people of 45-59 years. The change in CMR of people died with/of AMI caused by demographic factors was 28.70% (95% CI: 12.99,46.60, p = 0.001) and 28.07% (95% CI: 12.71,45.52, p = 0.001) per year, respectively. Conclusion: Preventative strategies for AMI should be applied to enhance the health management of residents aged 45-59 years or with comorbidities in the transitioning region.

The burden of congenital birth defects between 1990 and 2019 in China: an observational study.

Background: Congenital birth defects (CBDs) are a major public health issue. This study aims to assess trends in the burden of CBDs between 1990 and 2019 across China based on the Global Burden of Disease Study 2019 (GBD 2019). Methods: Indicators of the burden of CBDs included incidence, mortality, and disability-adjusted life years (DALYs). Metrics included number, rate, and age-standardized rate with 95% uncertainty intervals (UIs). Data were stratified by region [China, global, high-, middle-, low-socio-demographic index (SDI)], age, sex, and type of CBD. Average annual percentage changes (AAPC) and trends were evaluated. Results: In China, between 1990 and 2019, the age-standardized incidence rate for CBDs showed an increasing trend, with an AAPC of 0.26% (0.11% to 0.41%), reaching 148.12 per 105 person-years (124.03 to 176.33) in 2019. Most CBDs were congenital heart anomalies, with an AAPC of 0.12% (-0.08% to 0.32%). The age-standardized mortality rate for CBDs showed a decreasing trend, with an AAPC of -4.57% (-4.97% to -4.17%), reaching 4.62 per 105 person-years (3.88 to 5.57) in 2019. Most mortality was associated with congenital heart anomalies, with an AAPC of -3.77% (-4.35% to -3.19%). The age-standardized DALYs rate for CBDs showed a decreasing trend, with an AAPC of -3.74% (-3.95% to -3.52%), reaching 480.95 per 105 person-years (407.69 to 570.04) in 2019. Conclusions: Morbidity associated with CBDs increased in China between 1990 and 2019, accelerated by the adoption of the two-child policy, and ranked high globally. These findings emphasize the need for prenatal screening and primary and secondary prevention strategies.

KIN17 promotes cell migration and invasion through stimulating the TGF-ß/Smad2 pathway in hepatocellular carcinoma.

KIN17 DNA and RNA binding protein (Kin17) is involved in the regulation of tumorigenesis of diverse human cancers. However, its role in the cancer progression and metastasis in hepatocellular carcinoma (HCC) remains largely unknown. Bioinformatics and immunohistochemistry staining were used to investigate the expression pattern of KIN17 and its prognostic value in HCC patients. The transwell, wound-healing assay was employed to determine the effects of KIN17 on migration and invasion of HCC cells in vitro. The tail veins model was employed to determine the effects of KIN17 on lung metastasis in vivo. The biological mechanisms involved in cell migration and invasion regulated by KIN17 were determined with Western blot analysis method. KIN17 expression was significantly increased in HCC tissues compared with adjacent normal tissues, with particularly higher in portal vein tumor thrombus and intrahepatic metastasis tissues. Patients with higher KIN17 expression experienced poor overall and disease free survival. KIN17 knockdown in HuH7 and HepG2 cells significantly reduced cell migration and invasion abilities, whereas its overexpression promoted migration and invasion in MHCC-97L and HepG2 cells in vitro and in vivo. In HuH7 and HepG2 cells, KIN17 knockdown inhibited the TGF-ß/Smad2 pathway. In contrast, KIN17 overexpression stimulated TGF-ß/Smad2 pathway in MHCC-97L and HepG2 cells, along with the genes involved in the epithelial-mesenchymal transition. These findings suggest that KIN17 promotes migration and invasion in HCC cells by stimulating the TGF-ß/Smad2 pathway. KIN17 could be a promising prognostic biomarker, as well as a potential therapeutic target in HCC.

Identification of key candidate genes and biological pathways in neuropathic pain.

BACKGROUND: Neuropathic pain is a common chronic pain, characterized by spontaneous pain and mechanical allodynia. The incidence of neuropathic pain is on the rise due to infections, higher rates of diabetes and stroke, and increased use of chemotherapy drugs in cancer patients. At present, due to its pathophysiological process and molecular mechanism remaining unclear, there is a lack of effective treatment and prevention methods in clinical practice. Now, we use bioinformatics technology to integrate and filter hub genes that may be related to the pathogenesis of neuropathic pain, and explore their possible molecular mechanism by functional annotation and pathway enrichment analysis. METHODS: The expression profiles of GSE24982, GSE2884, GSE2636 and GSE30691 were downloaded from the Gene Expression Omnibus（GEO）database, and these datasets include 93 neuropathic pain Rattus norvegicus and 59 shame controls. After the four datasets were all standardized by quantiles, the differentially expressed genes (DEGs) between NPP Rattus norvegicus and the shame controls were finally identified by the robust rank aggregation (RRA) analysis method. In order to reveal the possible underlying biological function of DEGs, the Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway enrichment analysis of DEGs were performed. In addition, a Protein-protein Interaction (PPI) network was also established. At the end of our study, a high throughput sequencing dataset GSE117526 was used to corroborate our calculation results. RESULTS: Through RRA analysis of the above four datasets GSE24982, GSE2884, GSE2636, and GSE30691, we finally obtained 231 DEGs, including 183 up-regulated genes and 47 down-regulated genes. Arranging 231 DEGs in descending order according to |log2 fold change (FC)|, we found that the top 20 key genes include 14 up-regulated genes and 6 down-regulated genes. The most down-regulated hub gene abnormal expressed in NPP was Egf17 (P-value = 0.008), Camk2n2 (P-value = 0.002), and Lep (P-value = 0.02), and the most up-regulated hub gene abnormal expressed in NPP was Nefm (P-value = 1.08E-06), Prx (P-value = 2.68E-07), and Stip1 (P-value = 4.40E-07). In GO functional annotation analysis results, regulation of ion transmembrane transport (GO:0034765; P-value = 1.45E-09) was the most remarkable enriched for biological process, synaptic membrane (GO:0097060; P-value = 2.95E-08) was the most significantly enriched for cellular component, channel activity (GO:0015267; P-value = 2.44E-06) was the most prominent enriched for molecular function. In KEGG pathway enrichment analysis results, the top three notable enrichment pathways were Neuroactive ligand-receptor interaction (rno04080; P-value = 3.46E-08), Calcium signaling pathway (rno04020; P-value = 5.37E-05), and Osteoclast differentiation (rno04380; P-value = 0.000459927). Cav1 and Lep appeared in the top 20 genes in both RRA analysis and PPI analysis, while Nefm appeared in RRA analysis and datasets GSE117526 validation analysis, so we finally identified these three genes as hub genes. CONCLUSIONS: Our research identified the hub genes and signal pathways of neuropathic pain, enriched the pathophysiological mechanism of neuropathic pain to some extent, and provided a possible basis for the targeted therapy of neuropathic pain.

Golgi-localized calcium/manganese transporters FgGdt1 and FgPmr1 regulate fungal development and virulence by maintaining Ca2+ and Mn2+ homeostasis in Fusarium graminearum.

Calcium and manganese transporters play important roles in regulating Ca2+ and Mn2+ homeostasis in cells, which is necessary for the normal physiological activities of eukaryotes. Gdt1 and Pmr1 function as calcium/manganese transporters in the Golgi apparatus. However, the functions of Gdt1 and Pmr1 have not been previously characterized in the plant pathogenic fungus Fusarium graminearum. Here, we identified and characterized the biological functions of FgGdt1 and FgPmr1 in F. graminearum. Our study shows that FgGdt1 and FgPmr1 are both localized to the cis- and medial-Golgi. Disruption of FgGdt1 or FgPmr1 in F. graminearum caused serious defects in vegetative growth, conidiation, sexual development and significantly decreased virulence in wheat but increased deoxynivalenol (DON) production. Importantly, FgGdt1 is involved in Ca2+ and Mn2+ homeostasis and the severe phenotypic defects of the ΔFggdt1 mutant were largely due to loss of FgGdt1 function in Mn2+ transportation. FgGdt1-mCherry colocalizes with FgPmr1-GFP at the Golgi, and FgGDT1 exerts its biological function upstream of FgPMR1. Taken together, our results collectively demonstrate that the cis- and medial-Golgi-localized proteins FgGdt1 and FgPmr1 regulate Ca2+ and Mn2+ homeostasis of the Golgi apparatus, and this function is important in modulating the growth, development, DON biosynthesis and pathogenicity of F. graminearum.

Sensitization of Non-Small Cell Lung Cancer Cells to Gefitinib and Reversal of Epithelial-Mesenchymal Transition by Aloe-Emodin Via PI3K/Akt/TWIS1 Signal Blockage.

Objective: To explore the impacts of AE (aloe-emodin) in gefitinib-resistant NSCLC (non-small cell lung cancer) cells and the corresponding mechanism. Methods: PC9 and PC9-GR cells were cultured and treated by gefitinib, AE, or the combination of the two drugs. Then, viability, apoptosis, migration and invasion of cells were investigated using CCK-8, TUNEL, wound healing assay, and transwell assay, respectively. Female BALB/c nude mice were employed for the establishment of xenograft tumor models to examine the role of AE in tumor growth. Results: PC9-GR cells showed reduced apoptosis and enhanced cell viability, migration and invasion upon treatment by gefitinib, compared with PC9 cells. E-cahherin in PC9-GR cells was down-regulated, while Vimentin, Snail2 (or Slug) and Twist1 in PC9-GR cells were up-regulated, compared with PC9 cells. Meanwhile, treatment by a combination of gefitinib and AE significantly strengthened apoptosis of PC9-GR cells, while attenuated their migration and invasion, compared with the control group or treatment by gefitinib or AE alone. WB results showed that AE could reverse EMT and activation of PI3K/AKT signalling pathway in PC9-GR cells. In vivo experiments showed that tumor growth and EMT of PC9-GR cells were dramatically repressed after treatment by a combination of AE and gefitinib. Additionally, the use of SC97 (a PI3K/Akt pathway activator) could counteract the effects of AE in gefitinib-resistant PC9 cells. Conclusions: AE could enhance the gefitinib sensitivity of PC9-GR cells and reverse EMT by blocking PI3K/Akt/TWIS1 signal pathway.

Pterostilbene attenuates the proliferation and differentiation of TNF-α-treated human periodontal ligament stem cells.

Periodontitis is a common chronic inflammatory oral disease. The objective of periodontal treatment is to control infection whilst regenerating damaged periodontal tissue. The present study aimed to determine the potential effects of pterostilbene (PTE), a representative stilbene compound, on the proliferation and differentiation of human periodontal ligament stem cells (hPDLSCs). Different concentrations (1, 5, 10 and 20 µM) of PTE were applied to hPDLSCs, after which Cell Counting Kit-8 and western blotting assays were performed to examine the protein levels of Ki67, PCNA, p-IκBα, IκBα, Bcl-2, Bax, cleaved caspase3. The effect of PTE on the release of inflammatory factors, including IL-1ß, IL-6 and IL-10 was assessed by RT-qPCR. The apoptosis of TNF-α-induced hPDLSCs was evaluated by TUNEL assay and western blotting. Additionally, the role of PTE in hPDLSC mineralization was evaluated using alizarin red staining. The expression levels of mineralization indices, including RUNX2 and ALP were subsequently determined using western blotting. Subsequently, the target of PTE was predicted using TargetNet database and AutoDock v4.2 software and verified using western blotting. The results of the present study revealed that PTE promoted the proliferation of hPDLSCs in a concentration-dependent manner. Furthermore, PTE treatment decreased the release of inflammatory factors and alleviated the apoptosis of TNF-α-induced hPDLSCs. PTE was also demonstrated to promote the formation of mineral nodules in TNF-α-induced hPDLSCs. The Targetnet database, along with molecular docking, indicated that histone deacetylases (HDACs) were the probable targets of PTE upstream of regulating periodontitis. The results of western blotting implied that TNF-α significantly increased expression levels of HDAC2, 4, 6 and 8, whilst PTE treatment markedly decreased HDAC4, 6 and 8 expression in a concentration-dependent manner compared with the TNF-α group, which further confirmed these conclusions. In summary, results of the present study revealed that PTE promoted TNF-α-induced hPDLSC proliferation and differentiation, whilst alleviating inflammation and apoptosis. PTE also inhibited the expression of HDACs, which may be involved in the mechanism of periodontitis.

The molecular mechanism underlying pathogenicity inhibition by sanguinarine in Magnaporthe oryzae.

BACKGROUND: Sanguinarine (SAN) is a benzophenanthridine alkaloid that broadly targets a range of pathways in mammalian and fungal cells. In this study we set out to explore the molecular mechanism of sanguinarine inhibition of the fungal development and pathogenicity of Magnaporthe oryzae with the hope that sanguinarine will bolster the development of antiblast agents. RESULTS: We found that the fungus exhibited a significant reduction in vegetative growth and hyphal melanization while the spores produced long germ tubes on the artificial hydrophobic surface characteristic of a defect in thigmotropic sensing when exposed to 4, 8 and 0.5 µm sanguinarine, respectively. Consistent with these findings, we observed that the genes involved in melanin biosynthesis and the fungal hydrophobin MoMPG1 were remarkably suppressed in mycelia treated with 8 µm sanguinarine. Additionally, sanguinarine inhibited appressorium formation at a dose of 1.0 µm and this defect was restored by supplementing 5 mM of exogenous cAMP. By qRT-PCR assay we found cAMP pathway signalling genes such as MoCAP1 and MoCpkA were significantly repressed whereas MoCDTF1 and MoSOM1 were upregulated in sanguinarine-treated strains. Furthermore, we showed that sanguinarine does not selectively inhibit vegetative growth and appressorium formation of Guy11 but also other strains of M. oryzae. Finally, treatment of sanguinarine impaired the appressorium-mediated penetration and pathogenicity of M. oryzae in a dose-dependent manner. CONCLUSION: Based on our results we concluded that sanguinarine is an attractive antimicrobial candidate for fungicide development in the control of rice blast disease. © 2021 Society of Chemical Industry.

High-Quality Genome Assembly of Pseudopestalotiopsis theae, the Pathogenic Fungus Causing Tea Gray Blight.

Tea gray blight is one of the most serious foliar diseases of tea tree, caused by the plant-pathogenic fungus Pseudopestalotiopsis theae, which can affect production and quality of tea worldwide. We generated a highly contiguous, 50.41-Mbp genome assembly (N50 = 1.30 Mbp) of P. theae strain CYF27 by combining PacBio long-read and Illumina short-read sequencing technologies. We identified a total of 15,626 gene models, of which 1,038 genes encode putative secreted proteins. The high-quality genome assembly and annotation resource reported here will be useful for the study of fungal infection mechanisms and pathogen-host interaction.

Comparative assessment between bio-adhesive material and silk suture in regard to healing, clinical attachment level and width of keratinized gingiva in gingival recession defects cases.

The objective of this study was to compare the silk suture with a cyanoacrylate adhesive to stabilize the free gingival graft in conjunction with Er: YAG laser-assisted recipient site preparation to augment the keratinized tissue in gingival recession cases. This randomized trial comprised of 300 recession defects patients. All the included patients were diagnosed using Miller class I and II gingival recession defects classification. Group I sites were treated with a free gingival graft (FGG) harvested using an Er: YAG laser and further sutured with silk. Group II sites were stabilized with isoamyl 2 cyanoacrylate bio-adhesive material. Clinical parameters, such as gingival recession depth, clinical attachment level, gain in gingival tissue thickness, and width of keratinized gingiva were recorded at baseline, and at third month, sixth month, and 12th month postoperatively. The mean changes in gingival recession from months 3 to 6 and months and 6 to 12 were significant (p < 0.05) in both groups. However, the improvement in recession depth was better in group II than in group I. The mean change in clinical attachment level did not differ significantly between the groups at the different time intervals. However, values tended to be higher in group II than in group I. The width of the keratinized gingiva tended to be higher from baseline to 3 months, baseline to 6 months, baseline to 12 months, 3 to 6 months, and from 6 to 12 months in group II as compared with group I (p > 0.05). Cyanoacrylate could be used as a substitute to silk sutures to stabilize FGGs. Cyanoacrylate was easy to apply, consumed less operating time, and was considered equally efficacious for stabilizing FGGs.

Fusarium verticillioides FvPex8 Is a Key Component of the Peroxisomal Docking/Translocation Module That Serves Important Roles in Fumonisin Biosynthesis but Not in Virulence.

Peroxisomes are ubiquitous organelles in eukaryotes that fulfill various important metabolic functions. In this study, we investigated the role of docking/translocation module (DTM) peroxins, mainly FvPex8, FvPex13, FvPex14, and FvPex33, in Fusarium verticillioides development, virulence, and fumonisin B1 (FB1) biosynthesis. Protein interaction experiments suggested that FvPex13 serves as the central DTM subunit in F. verticillioides. Notably, FvPex8 and FvPex14 did not show direct interaction in our experiments. We generated gene-deletion mutants (ΔFvpex8, ΔFvpex13, ΔFvpex14, ΔFvpex33, ΔFvpex33/14) and further examined the functional role of these peroxins. Deletion mutants exhibited disparity in carbon nutrient utilization and defect in cell-wall integrity when stress agents were applied. Under nutrient starvation, mutants also showed higher levels of lipid droplet accumulation. Particularly, ΔFvpex8 mutant showed significant FB1 reduction and altered expression of key FB1 biosynthesis genes. However, FvPex13 was primarily responsible for asexual conidia reproduction and virulence, while the ΔFvpex33/14 double mutant also showed a virulence defect. In summary, our study suggests that FvPex13 is the central component of DTM, with direct physical interaction with other DTM peroxins, and regulates peroxisome membrane biogenesis as well as PTS1- and PTS2-mediated transmembrane cargo transportation. Importantly, we also characterized FvPex8 as a key component in F. verticillioides DTM that affects peroxisome function and FB1 biosynthesis.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Microbial community associated with ectomycorrhizal Russula symbiosis and dominated nature areas in southern China.

Russula griseocarnosa is one of the uncultivable important mycorrhizal edible fungi. Currently, there is a limited insight into the dynamic composition of the microbial communities associated with Russula. Here, the microbiota in the root and mycorrhizosphere from Russula-Fagaceae nature areas of Fujian province were identified by Illumina MiSeq high-throughput sequencing. First, we compared three types of fungal communities associated with Russula-Fagaceae root mycelia-running stage (stage-1), Russula sporocarping stage-2 (stage-2) and Russula-free Fagaceae root (stage-3). Fungal diversity negatively correlated with Russula. Russula, Tomentella and Lactarius were core EcM in Fagaceae roots. A total of eight genera, including Boletus, are likely a positive indicator of Russula sporocarp production in Russula-Fagaceae roots, while Tomentella and Elaphomyces for Russula symbiosis. Secondly, analysis of fungal and bacterial communities within rhizosphere soils from the three stages revealed six genera, including Dacryobolus and Acidocella, as possible indicator species associated with sporocarping in Russula. Elaphomyces, Tomentella, Sorangium, Acidicaldus, Acidobacterium and Haliangium occurred more frequently in the Russula rhizosphere. Furthermore, operational taxonomic unit (OTU) network analysis showed a positive correlation between Russula,Tomentella, Elaphomyces and Sorangium. Overall, our results revealed a relationship between micro-community and Russula, which may provide a new strategy for improving Russula symbiosis and sporocarp production.

The retromer CSC subcomplex is recruited by MoYpt7 and sequentially sorted by MoVps17 for effective conidiation and pathogenicity of the rice blast fungus.

In eukaryotic cells, Rab GTPases and the retromer complex are important regulators of intracellular protein transport. However, the mechanistic relationship between Rab GTPases and the retromer complex in relation to filamentous fungal development and pathogenesis is unknown. In this study, we used Magnaporthe oryzae, an important pathogen of rice and other cereals, as a model filamentous fungus to dissect this knowledge gap. Our data demonstrate that the core retromer subunit MoVps35 interacts with the Rab GTPase MoYpt7 and they colocalize to the endosome. Without MoYpt7, MoVps35 is mislocalized in the cytoplasm, indicating that MoYpt7 plays an important role in the recruitment of MoVps35. We further demonstrate that the expression of an inactive MoYpt7-DN (GDP-bound form) mutant in M. oryzae mimicks the phenotype defects of retromer cargo-sorting complex (CSC) null mutants and blocks the proper localization of MoVps35. In addition, our data establish that MoVps17, a member of the sorting nexin family, is situated at the endosome independent of retromer CSC but regulates the sorting function of MoVps35 after its recruitment to the endosomal membrane by MoYpt7. Taken together, these results provide insight into the precise mechanism of retromer CSC recruitment to the endosome by MoYpt7 and subsequent sorting by MoVps17 for efficient conidiation and pathogenicity of M. oryzae.

Improving the hepatitis cascade: assessing hepatitis testing and its management in primary health care in China.

Objective: The study aimed to decentralize hepatitis testing and management services to primary care in China. Methods: A nationwide representative provider survey amongst community health centres (CHCs) using randomized stratified sampling methods was conducted between September and December 2015. One hundred and eighty CHCs and frontline primary care practitioners from 20 cities across three administrative regions of Western, Central and Eastern China were invited to participate. Results: One hundred and forty-nine clinicians-in-charge (79%), 1734 doctors and 1846 nurses participated (86%). Majority of CHCs (80%, 95% CI: 74-87) offered hepatitis B testing, but just over half (55%, 95% CI: 46-65) offered hepatitis C testing. The majority of doctors (87%) and nurses (85%) felt that there were benefits for providing hepatitis testing at CHCs. The major barriers for not offering hepatitis testing were lack of training (54%) and financial support (23%). Multivariate analysis showed that the major determinants for CHCs to offer hepatitis B and C testing were the number of nurses (AOR 1.1) and written policies for hepatitis B diagnosis (AOR 12.7-27.1), and for hepatitis B the availability of reproductive health service. Conclusions: Primary care providers in China could play a pivotal role in screening, diagnosing and treating millions of people with chronic hepatitis B and C in China.

Opportunities and barriers to STI testing in community health centres in China: a nationwide survey.

BACKGROUND: China has strengthened its primary care workforce and implemented a wide network of community health centres (CHCs). However, STI testing and management are not currently included in the 'Essential Package of Primary Health Care in China'. Legislation change to encourage STI service delivery would be important, but it is also critical to determine if there are also provider-related opportunities and barriers for implementing effective STI programmes through CHCs if future legislation were to change. METHODS: A national representative survey was conducted between September and December 2015 in a stratified random sample of 180 CHCs based in 20 cities in China. Primary care practitioners (PCPs) provided information on current experiences of STI testing as well as the barriers and facilitators for STI testing in CHCs. Multivariate logistic regression was conducted to determine factors associated with PCPs performing STI testing. RESULTS: 3580 out of 4146 (86%) invited PCPs from 158 CHCs completed the survey. The majority (85%, 95% CI 84% to 87%) of doctors stated that STI testing was an important part of healthcare. However, less than a third (29%, 95% CI 27% to 31%) would perform an STI test if the patients asked. Barriers for performing STI testing included lack of training, concerns about reimbursement, concerns about damage to clinics' reputations and the stigma against key populations. Respondents who reported that they would perform an STI test were likely to be younger, received a bachelor degree or higher, received specific training in STIs, believed that STI test was an important part of healthcare or had resources to perform STI testing. CONCLUSIONS: There is potential for improving STI management in China through upskilling the primary care workforce in CHCs. Specific training in STIs is needed, and other structural, logistical and attitudinal barriers are needed to be addressed.

Primary care workforce and continuous medical education in China: lessons to learn from a nationwide cross-sectional survey.

OBJECTIVES: This study aimed to examine the education and training background of Chinese community health centres (CHCs) staff, continuous medical education (CME) and factors affecting participation in CME. DESIGN: Cross-sectional survey. SETTING: Community health centres (CHCs). PARTICIPANTS: All doctors and nurses working in selected CHCs (excluding those solely practising traditional Chinese Medicine). MAIN OUTCOME MEASURES: CME recorded by CHCs and self-reported CME participation. METHODS: A stratified random sample of CHCs based on geographical distribution and 2:1 urban-suburban ratio was selected covering three major regions of China. Two questionnaires, one for lead clinicians and another for frontline health professionals, were administered between September-December 2015, covering the demographics of clinic staff, staff training and CME activities. RESULTS: 149 lead clinicians (response rate 79%) and 1734 doctors and 1846 nurses completed the survey (response rate 86%). Of the doctors, 54.5% had a bachelor degree and only 47% were registered as general practitioners (GPs). Among the doctors, 10.5% carried senior titles. Few nurses (4.6%) had training in primary care. Those who have reported participating in CME were 91.6% doctors and 89.2% nurses. CME participation in doctors was more commonly reported by older doctors, females, those who were registered as a GP and those with intermediate or senior job titles. CME participation in nurses was more common among those with a bachelor degree or intermediate/senior job titles or those with longer working experience in the CHC. CONCLUSION: Only half of doctors have bachelor degrees or are registered as GPs as their prime registration in the primary care workforce in China. The vast majority of CHC staff participated in CME but there is room for improvement in how CME is organised.

Bridging the Gaps Between Patients and Primary Care in China: A Nationwide Representative Survey.

PURPOSE: China introduced a national policy of developing primary care in 2009, establishing 8,669 community health centers (CHCs) by 2014 that employed more than 300,000 staff. These facilities have been underused, however, because of public mistrust of physicians and overreliance on specialist care. METHODS: We selected a stratified random sample of CHCs throughout China based on geographic distribution and urban-suburban ratios between September and December 2015. Two questionnaires, 1 for lead clinicians and 1 for primary care practitioners (PCPs), asked about the demographics of the clinic and its clinical and educational activities. Responses were obtained from 158 lead clinicians in CHCs and 3,580 PCPs (response rates of 84% and 86%, respectively). RESULTS: CHCs employed a median of 8 physicians and 13 nurses, but only one-half of physicians were registered as PCPs, and few nurses had training specifically for primary care. Although virtually all clinics were equipped with stethoscopes (98%) and sphygmomanometers (97%), only 43% had ophthalmoscopes and 64% had facilities for gynecologic examination. Clinical care was selectively skewed toward certain chronic diseases. Physicians saw a median of 12.5 patients per day. Multivariate analysis showed that more patients were seen daily by physicians in CHCs organized by private hospitals and those having pharmacists and nurses. CONCLUSIONS: Our survey confirms China's success in establishing a large, mostly young primary care workforce and providing ongoing professional training. Facilities are basic, however, with few clinics providing the comprehensive primary care required for a wide range of common physical and mental conditions. Use of CHCs by patients remains low.

Characterization of the Adult Head Transcriptome and Identification of Migration and Olfaction Genes in the Oriental Armyworm Mythimna separate.

The oriental armyworm Mythimna separate is an economically important insect with a wide distribution and strong migratory activity. However, knowledge about the molecular mechanisms regulating the physiological and behavioural responses of the oriental armyworm is scarce. In the present study, we took a transcriptomic approach to characterize the gene network in the adult head of M. separate. The sequencing and de novo assembly yielded 63,499 transcripts, which were further assembled into 46,459 unigenes with an N50 of 1,153 bp. In the head transcriptome data, unigenes involved in the 'signal transduction mechanism' are the most abundant. In total, 937 signal transduction unigenes were assigned to 22 signalling pathways. The circadian clock, melanin synthesis, and non-receptor protein of olfactory gene families were then identified, and phylogenetic analyses were performed with these M. separate genes, the model insect Bombyx mori and other insects. Furthermore, 1,372 simple sequence repeats of 2-6 bp in unit length were identified. The transcriptome data represent a comprehensive molecular resource for the adult head of M. separate, and these identified genes can be valid targets for further gene function research to address the molecular mechanisms regulating the migratory and olfaction genes of the oriental armyworm.