Exploring the neural mechanisms underlying achalasia: A study of functional connectivity and regional brain activity.

BACKGROUND AND AIMS: The pathophysiology of achalasia, which involves central nuclei abnormalities, remains unknown. We investigated the resting-state functional MRI (rs-fMRI) features of patients with achalasia. METHODS: We applied resting-state functional MRI (rs-fMRI) to investigate the brain features in patients with achalasia (n = 27), compared to healthy controls (n = 29). Focusing on three regions of interest (ROIs): the dorsal motor nucleus of the vagus (DMV), the nucleus ambiguus (NA), and the nucleus of the solitary tract (NTS), we analyzed variations in resting-state functional connectivity (rs-FC), fractional amplitude of low-frequency fluctuations (fALFF), and regional homogeneity (ReHo). RESULTS: Achalasia patients demonstrated stronger functional connectivity between the NA and the right precentral gyrus, left postcentral gyrus, and left insula. No significant changes were found in the DMV or NTS. The fMRI analysis showed higher rs-FC values for NA-DMV and NA-NTS connections in achalasia patients. Achalasia patients exhibited decreased fALFF values in the NA, DMV, and NTS regions, as well as increased ReHo values in the NA and DMV regions. A positive correlation was observed between fALFF values in all six ROIs and the width of the barium meal. The NTS fALFF value and NA ReHo value displayed a positive correlation with integrated relaxation pressure (IRP), while the ReHo value in the right precentral gyrus showed an inverse correlation with the height of the barium meal. CONCLUSIONS: Abnormal rs-FC and regional brain activity was found in patients with achalasia. Our study provides new insights into the pathophysiology of achalasia and highlights the potential of rs-fMRI in improving the diagnosis and treatment of this condition.

Differences and biocontrol potential of haustorial endophytic fungi from Taxillus Chinensis on different host plants.

BACKGROUND: To explore the community composition and diversity of the endophytic fungi in Taxillus chinensis, samples of the parasites growing on seven different hosts, Morus alba, Prunus salicina, Phellodendron chinense, Bauhinia purpurea, Dalbergia odorifera, Diospyros kaki and Dimocarpus longan, were isolated. The strains were identified by their morphological characteristics and their internal transcribed spacer (ITS) sequences. RESULTS: 150 different endophytic fungi were isolated from the haustorial roots of the seven hosts with a total isolation rate of 61.24%. These endophytic fungi were found to belong to 1 phylum, 2 classes, 7 orders, 9 families, 11 genera and 8 species. Among of them, Pestalotiopsis, Neopestalotiopsis and Diaporthe were the dominant genera, accounting for 26.67, 17.33 and 31.33% of the total number of strains, respectively. Diversity and similarity analyses showed that the endophytic fungi isolated from D. longan (H'=1.60) had the highest diversity index. The highest richness indexes were found in M. alba and D. odorifera (both 2.23). The evenness index of D. longan was the highest (0.82). The similarity coefficient of D. odorifera was the most similar to D. longan and M. alba (33.33%), while the similarity coefficient of P. chinense was the lowest (7.69%) with M. alba and D. odorifera. Nine strains showed antimicrobial activities. Among them, Pestalotiopsis sp., N. parvum and H. investiens showed significant antifungal activity against three fungal phytopathogens of medicinal plants. At the same time, the crude extracts from the metabolites of the three endophytic fungi had strong inhibitory effects on the three pathogens. Pestalotiopsis sp., N. parvum and H. investiens had the strongest inhibitory effects of S. cucurbitacearum, with inhibitory rates of 100%, 100% and 81.51%, respectively. In addition, N. parvum had a strong inhibitory effect on D. glomerata and C. cassicola, with inhibitory rates of 82.35% and 72.80%, respectively. CONCLUSIONS: These results indicate that the species composition and diversity of endophytic fungi in the branches of T. chinensis were varied in the different hosts and showed good antimicrobial potential in the control of plant pathogens.

X-ray microscopy enables multiscale high-resolution 3D imaging of plant cells, tissues, and organs.

Capturing complete internal anatomies of plant organs and tissues within their relevant morphological context remains a key challenge in plant science. While plant growth and development are inherently multiscale, conventional light, fluorescence, and electron microscopy platforms are typically limited to imaging of plant microstructure from small flat samples that lack a direct spatial context to, and represent only a small portion of, the relevant plant macrostructures. We demonstrate technical advances with a lab-based X-ray microscope (XRM) that bridge the imaging gap by providing multiscale high-resolution three-dimensional (3D) volumes of intact plant samples from the cell to the whole plant level. Serial imaging of a single sample is shown to provide sub-micron 3D volumes co-registered with lower magnification scans for explicit contextual reference. High-quality 3D volume data from our enhanced methods facilitate sophisticated and effective computational segmentation. Advances in sample preparation make multimodal correlative imaging workflows possible, where a single resin-embedded plant sample is scanned via XRM to generate a 3D cell-level map, and then used to identify and zoom in on sub-cellular regions of interest for high-resolution scanning electron microscopy. In total, we present the methodologies for use of XRM in the multiscale and multimodal analysis of 3D plant features using numerous economically and scientifically important plant systems.

The effects of the prognostic biomarker SAAL1 on cancer growth and its association with the immune microenvironment in lung adenocarcinoma.

BACKGROUND: Inhibition of Serum Amyloid A-like 1 (SAAL1) expression could inhibit cancer progression and improve the prognosis of cancer patients. At present, the correlation between SAAL1 and lung adenocarcinoma (LAC) remains unclear. Therefore, this study surveyed the worth and pathway of SAAL1 in LAC progression and immunity. METHODS: Bioinformatics and immunohistochemistry were used to identify the SAAL1 expression in LAC. The roles of SAAL1 expression in the existence values of LAC patients were explored, and the nomograms were constructed. Clinical values of SAAL1 co-expressed genes were evaluated by COX regression, survival, and Receiver operating characteristic (ROC) analysis. EDU and western blotting methods were used to inquiry the functions and pathways of the SAAL1 in cell growths. The correlation between the SAAL1 level and immune microenvironment was visualized using correlation research. RESULTS: SAAL1 level was elevated in LAC tissues, and was observed in cancer tissues of dead patients. SAAL1 overexpression had something to do with shorter overall survival, progression-free interval, and disease-specific survival in LAC. The area under the curve of SAAL1 was 0.902 in normal tissues and cancer tissues. Inhibition of SAAL1 expression could inhibit cancer cell proliferation, which may be related to the decreased expression of cyclin D1 and Bcl-2 proteins. In LAC, SAAL1 level had something to do with stromal, immune, and estimate scores, and correlated with macrophages, T cells, Th2 cells, CD8 T cells, NK CD56dim cells, DC, eosinophils, NK CD56bright cells, pDC, iDC, cytotoxic cells, Tgd, aDC cells, B cells, Tcm, and TFH levels. SAAL1 overexpression had something to do with existence values and the immunity in LAC. CONCLUSIONS: Inhibition of SAAL1 expression could regulate cancer growth via cyclin D1 and Bcl-2. SAAL1 is a promising prognostic biomarker in LAC patients.

Overexpression of SH2D1A promotes cancer progression and is associated with immune cell infiltration in hepatocellular carcinoma via bioinformatics and in vitro study.

BACKGROUND: SH2 domain containing 1A (SH2D1A) expression has been linked to cancer progression. However, the functions of SH2D1A in hepatocellular carcinoma (HCC) have not been reported. METHODS: The effects of SH2D1A on the proliferation, migration, and invasion of HCC cells and the related pathways were re-explored in cell models with SH2D1A overexpression using the CCK-8, migration and invasion assays and western blotting. The functions and mechanisms of genes co-expressed with SH2D1A were analyzed using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The relationship between SH2D1A expression and immune microenvironment features in HCC was explored. RESULTS: Elevated SH2D1A expression promoted cell proliferation, migration, and invasion, which was related to the overexpression of p-Nf-κB and BCL2A1 protein levels in HCC. SH2D1A expression was related to the immune, stromal, and ESTIMATE scores, and the abundance of immune cells, such as B cells, CD8+ T cells, and T cells. SH2D1A expression was significantly related to the expression of immune cell markers, such as PDCD1, CD8A, and CTLA4 in HCC. CONCLUSION: SH2D1A overexpression was found to promote cell growth and metastasis via the Nf-κB signaling pathway and may be related to the immune microenvironment in HCC. The findings indicate that SH2D1A can function as a biomarker in HCC.

CLDN4 as a Novel Diagnostic and Prognostic Biomarker and Its Association with Immune Infiltrates in Ovarian Cancer.

Ovarian cancer (OC) is the seventh most prevalent type of cancer in women and the second most common cause of cancer-related deaths in women worldwide. Because of the high rates of relapse, there is an immediate and pressing need for the discovery of innovative sensitive biomarkers for OC patients. Using TCGA and GSE26712 datasets, we were able to identify 17 survival-related DEGs in OC that had differential expression. CLDN4 was the gene that caught our attention the most out of the 17 important genes since its expression was much higher in OC samples than in nontumor samples. The findings of the ROC assays then confirmed the diagnostic utility of the test in screening OC specimens to differentiate them from nontumor specimens. Patients with high CLDN4 expression predicted a shorter overall survival (OS) and disease-specific survival (DSS) than those with low CLDN4 expression, according to clinical research. Patients with low CLDN4 expression predicted longer OS and DSS. Analysis using both univariate and multivariate techniques revealed that CLDN4 expression was an independent factor associated with a poor prognosis for OS and DSS. Based on multivariate analysis, the C-indexes and calibration plots of the nomogram suggested an effective predictive performance for OC patients. After that, we investigated whether or not there was a link between the infiltration of immune cells and the expression of the CLDN4 gene. We found that the expression of CLDN4 was positively associated with Th17 cells, NK CD56bright cells, while negatively associated with Th2 cells, pDC, and T helper cells. In the end, we carried out RT-PCR on our cohort and confirmed that the level of CLDN4 expression was noticeably elevated in OC specimens in comparison to nontumor tissues. The diagnostic usefulness of CLDN4 expression for OC was also validated by the findings of ROC tests. Thus, our findings revealed that CLDN4 may serve as a predictive biomarker in OC to assess both the clinical outcome of OC patients and the level of immune infiltration.

Awareness of HPV and HPV vaccines, acceptance to vaccination and its influence factors among parents of adolescents 9 to 18 years of age in China: A cross-sectional study.

BACKGROUND: Vaccination uptake rates for adolescents are still low in China despite safe and effective human papillomavirus vaccines being available. The awareness and attitudes of parents to HPV vaccines play a decisive role in adolescents' HPV vaccination uptake. METHODS: A cross-sectional study was conducted from March, 2022 to May, 2022 using an anonymous questionnaire among parents of 9 to 18 years of age from 73 cities in 23 provinces in mainland China. Demographic characteristics of parents, their knowledge and attitudes about HPV and HPV vaccination, as well as factors influencing HPV vaccination in adolescents were assessed. RESULTS: More than two-thirds of parents heard of HPV (75.5%) and HPV vaccines (84.7%). Of these participants, mothers (83.8%) were in the majority. Parents willing to vaccinate themselves and their children against HPV were 84.9% and 87.6%, respectively. Parents were more likely to vaccinate their daughters against HPV than their sons (P < 0.001). Parents who had heard of the HPV vaccines (P = 0.028) or had vaccinated themselves (P < 0.001) were more likely to have HPV vaccination for their children. Parents who accepted the price of the HPV vaccines (P = 0.005) were more likely to have their children vaccinated against HPV. CONCLUSIONS: Children's gender, awareness of the HPV vaccines, parental HPV vaccination, and the price of the HPV vaccines are likely to be the reason for parents' vaccine hesitancy for adolescents. PRACTICE IMPLICATIONS: Nurses have a critical role in identifying parental hesitancy and providing individualized education to expand the parental awareness and knowledge and encourage on-time adolescents vaccination.

Three-Dimensional Time-Lapse Analysis Reveals Multiscale Relationships in Maize Root Systems with Contrasting Architectures.

Understanding how an organism's phenotypic traits are conditioned by genetic and environmental variation is a central goal of biology. Root systems are one of the most important but poorly understood aspects of plants, largely due to the three-dimensional (3D), dynamic, and multiscale phenotyping challenge they pose. A critical gap in our knowledge is how root systems build in complexity from a single primary root to a network of thousands of roots that collectively compete for ephemeral, heterogeneous soil resources. We used time-lapse 3D imaging and mathematical modeling to assess root system architectures (RSAs) of two maize (Zea mays) inbred genotypes and their hybrid as they grew in complexity from a few to many roots. Genetically driven differences in root branching zone size and lateral branching densities along a single root, combined with differences in peak growth rate and the relative allocation of carbon resources to new versus existing roots, manifest as sharply distinct global RSAs over time. The 3D imaging of mature field-grown root crowns showed that several genetic differences in seedling architectures could persist throughout development and across environments. This approach connects individual and system-wide scales of root growth dynamics, which could eventually be used to predict genetic variation for complex RSAs and their functions.

Successful treatment of two patients with unresectable lung squamous cell carcinoma with tislelizumab regardless of programmed death-ligand 1 expression: a report of two cases.

Since the treatment of lung squamous cell carcinoma (SCC) was limited due to a lack of appropriate biomarkers and novel target agents. Immune checkpoint inhibitors can offer an effective treatment for patients with advanced non-small cell lung cancer. Here, we described the cases of two patients with SCC who showed a good response following treatment with tislelizumab. We encountered two patients with unresectable lung SCC who were treated with immunotherapy and chemotherapy. One patient had negatively programmed death-ligand 1 expression, and the primary lesion becomes a thick wall cavity after the tislelizumab combined with chemotherapy. Another patient was diagnosed with advanced lung SCC with negative programmed death-ligand 1 expression. After the treatment, the fluorine-18-fluorodeoxyglucose PET/computed tomography indicated that no abnormal increase in radioactivity uptake and tend to complete remission. We found a significant response or even complete response in unresectable SCC treated with tislelizumab combined with chemotherapy. Our cases added evidence of the feasibility and efficacy of tislelizumab combined with chemotherapy in unresectable lung SCC.

Diagnostic Value of FibroTouch and Non-invasive Fibrosis Indexes in Hepatic Fibrosis with Different Aetiologies.

BACKGROUND: Liver biopsy is the gold standard for staging liver fibrosis, but it has numerous drawbacks, mainly associated with bleeding and bile fistula risks. A number of non-invasive techniques have been investigated, but they all have their own disadvantages. To avoid the risks mentioned above and to improve the diagnostic value, we still need to search for a more accurate non-invasive method to evaluate the degree of liver fibrosis. AIM: This study aimed to evaluate the diagnostic performance of FibroTouch versus other non-invasive fibrosis indexes in hepatic fibrosis of different aetiologies. METHODS: This study retrospectively enrolled 227 patients with chronic hepatic liver disease admitted to the first hospital of Lanzhou University from 2017 to 2020. Liver biopsy was performed in all of the patients, and their biochemical indicators were all tested. Non-invasive indexes including the fibrosis index based on four factors (FIB-4), the aminotransferase-to-platelet ratio index (APRI), and the gamma-glutamyl transpeptidase-to-platelet ratio index (GPRI) were all calculated. Transient elastography was performed using FibroTouch. RESULTS: The correlation between FibroTouch and the pathology of liver fibrosis was significantly higher than that between the non-invasive fibrosis indexes and the biopsy results (r = 0.771, p < 0.05). The area under the receiver operating curve (AUC) of FibroTouch was significantly higher than that of FIB-4, APRI, and GPRI for the diagnosis of significant fibrosis (≥ S2 fibrosis stage), advanced fibrosis (≥ S3 fibrosis stage), and cirrhosis (= S4 fibrosis stage) (p < 0.05). The patients were grouped according to different aetiologies. The diagnostic value of FibroTouch had much higher credibility in different fibrosis stages for different causes compared with other non-invasive indexes. The AUC of FibroTouch showed both higher specificity and higher sensitivity than FIB-4, APRI, and GPRI for different liver fibrosis stages with different aetiologies. CONCLUSIONS: FibroTouch demonstrates the highest diagnostic value for liver fibrosis and cirrhosis among non-invasive methods, showing better results than FIB-4, APRI, and GPRI, and surpassed only by liver biopsy. FibroTouch is reliable in assessing liver fibrosis with different aetiologies.

Prevalence, correlates, and trajectory of screen viewing among Chinese children in Changsha: a birth cohort study.

BACKGROUND: High screen viewing time has detrimental effects on children's health, development, and behavior developing. Children are being exposed to more and more media devices at an earlier age. This study was aimed to determine the amount of daily screen time and its variation and to assess potential factors of screen time by identifying the trajectory of screen time among children aged 1 to 5 years. METHOD: This study was based on a representative sample of Changsha young children from a cohort study during 2015-2020. The demographic information and children's screen viewing time were collected by parents or caregivers through face-to-face interviews. The Latent growth model was used to test the effects of outdoor play on screen viewing time at eight time points, meanwhile, unconditional and conditional models were examined sequentially. RESULT: After excluding respondents with missing key variables, we included 953 children in the final analysis. Children's outdoor play was slightly increased at 18 months and subsequently declined at 24-60 months, with a maximum duration of 2.96 h per day. Children's average screen time was increased at 18-36 months, and decreased at 42-54 months, with a slight increase at 60 months. The duration of media exposure peaked at 1.4 h/d at age of 36 months and 60 months. Standardized coefficients of the outdoor play at age of 12 months showed negative effects on the screen time in children, but with positive influence at age of 24, 36, and 42 months (P <  0.01). CONCLUSION: High proportions of young Chinese children in Changsha had more screen time than the AAP recommended according to our analysis. Significant predictors of screen time included pregnancy computer use, paternal educational level, and outdoor play in this study, however, further understanding of risk factors is needed to promote great public health efforts to reduce children's screen exposure.

NGF/TrkA promotes the vitality, migration and adhesion of bone marrow stromal cells in hypoxia by regulating the Nrf2 pathway.

BACKGROUND: Bone marrow stromal cells (BMSCs) transplantation is a treatment strategy for ischemic stroke (IS) with great potential. However, the vitality, migration and adhesion of BMSCs are greatly impaired due to the harsh environment of the ischemic area, which affects the therapeutic effects. Herein, we aimed to investigate the roles of nerve growth factor (NGF) in regulating cell behaviors of BMSCs in IS. METHODS: The mRNA and protein expressions were assessed using qRT-PCR and western blot, respectively. To simulate ischemic-like conditions in vitro, Brain microvascular (bEnd.3) cells were exposed to oxygen and glucose deprivation (OGD). Cell viability and cell proliferation were evaluated by MTT assay and BrdU assay, respectively. Transwell migration and cell adhesion assays were carried out to determine cell migration and adhesion of BMSCs, respectively, coupled with flow cytometry to evaluate cell apoptosis of bEnd.3 cells. Finally, angiogenesis assay was performed to assess the angiogenesis ability of bEnd.3 cells. RESULTS: NGF overexpression resulted in increased cell vitality, adhesion and migration of BMSCs, while NGF knockdown presented the opposite effects. We subsequently discovered that TrkA was a receptor for NGF, and TrkA knockdown significantly inhibited the cell viability, migration and adhesion of BMSCs. Besides, Nrf2 was confirmed as the downstream target of NGF/TrkA to promote the viability, adhesion and migration of BMSC cells. Finally, NGF-silenced BMSCs could not effectively restore the OGD-induced brain microvascular cell damage. CONCLUSIONS: NGF/TrkA promoted the viability, migration and adhesion of BMSCs in IS via activating Nrf2 pathway.

Association of outdoor activity time and screen time with sleep for 36 to 54 months children in Kaifu District, Changsha: A birth cohort study. / 长沙市开福区36~54æœˆé¾„å„¿ç«¥æˆ·å¤–æ´»åŠ¨æ—¶é—´å’Œå±å¹•æ—¶é—´ä¸Žç¡çœ å ³ç³»çš„å‡ºç”Ÿé˜Ÿåˆ—ç ”ç©¶.

OBJECTIVES: The prevalence rate of sleep problems in children in China is increasing in recent years. There are inconsistencies in the relationship between physical activity and sleep, and the impact of screen time on young children seems more obvious.This study aims to understand the current situation of outdoor activity time, screen time and sleep (total sleep duration, bedtime, and wake-up time) and the associations between outdoor activity time and screen time with sleep for children aged 36-54 months in Kaifu District of Changsha. METHODS: Using the cluster sampling method, 1 286 newborns delivered in 3 community health service centers in Kaifu District, Changsha from January to December 2015 were selected as the research subjects to establish a prospective birth cohort. According self-designed questionnaire and household follow-up to select 36, 42, 48, 54 months Children's data. The mixed linear model was used to explore the associations of outdoor activity time and screen time with sleep. RESULTS: The 36-54 months children's total sleep duration was decreased from 11.60 h/d to 10.92 h/d (P<0.001); bedtime time delayed from 21:58 to 22:00 (P=0.124); wake-up time advanced from 7:52 to 7:37 (P<0.001); outdoor activity time was decreased from 2.58 h/d to 1.94 h/d (P<0.001), and screen time was decreased from 1.28 h/d to 1.09 h/d (P<0.001). With aging, the prevalence of sleep <10 h/d and outdoor activity time <2 h/d was increased significantly, and the prevalence of screen time ≥1 h/d was decreased (P<0.05). Mixed linear models showed that longer outdoor activity time was related to increased total sleep duration (ß=0.22, 95% CI 0.00 to 0.44) and delayed wake-up time (ß=0.16, 95% CI 0.00 to 0.32), and longer screen time was related to delayed bedtime (ß=0.22, 95% CI 0.05 to 0.39) and wake-up time (ß=0.24, 95% CI 0.08 to 0.41). These associations were different in gender. Longer outdoor activity time was related to delayed wake-up time (ß=0.37, 95% CI 0.14 to 0.59) in boys, but not in girls (ß=-0.16, 95% CI -0.33 to 0.01). Longer screen time was associated with delayed bedtime (ß=0.40, 95% CI 0.09 to 0.63) and wake-up time (ß=0.33, 95% CI 0.15 to 0.51) in girls, but only related to delayed wake-up time (ß=0.29, 95% CI 0.06 to 0.52) in boys, and the degree of association was lower than that of girls. CONCLUSIONS: Among children aged 36-54 months in Kaifu District, Changsha, there are problems including going to bed late and getting up late, insufficient physical activity, and long screen time. Outdoor activity time and screen time are related to sleep. Increasing outdoor activity time and reducing screen time can help to improve children's sleep.

Morphology and activities of cell populations of haemocytes in Oncomelania hupensis following Schistosoma japonicum infection.

Oncomelania hupensis is the only obligatory intermediate host of Schistosoma japonicum, the pathogen of zoonosis schistosomiasis. Haemocytes play a critical role in the cellular immune defence of O. hupensis against S. japonicum challenge. Here, the morphology and classification of haemocytes of O. hupensis were investigated by Giemsa staining and light microscopy, combining with the scanning and transmission electron microscopy and flow cytometry. Granulocytes and hyalinocytes were confirmed as two main types of haemocytes, account for ~ 10% and ~ 90% of all haemocytes, with size varying in 4.3-10.9 µm and 0.4-30.8 µm, respectively. Subpopulations can be identified further by granule feature, shape, size, and surface and inner structure of cells. The heterogeneity in morphology implied varied developmental process and function of haemocyte subpopulations. After the S. japonicum challenge, haemocytes of O. hupensis respond to S. japonicum invasion immediately. The dynamic change of haemocyte subpopulations indicates that the small hyalinocyte could differentiate into a larger one or granulocyte after S. japonicum challenge, and the granulocytes and larger hyalinocytes play leading roles in early defence reaction, but in different ways. Phagocytosis and apoptosis of haemocytes in O. hupensis were proved to be related to immune defence against S. japonicum, with the combined effect of granulocytes and larger hyalinocytes. However, the main pathway of each subpopulation to take effect in different periods need further investigation.

A large-scale systematic framework of Chinese snakes based on a unified multilocus marker system.

Snakes are one of the most diverse groups of terrestrial vertebrates, with approximately 3500 extant species. A robust phylogeny and taxonomy of snakes is crucial for us to know, study and protect them. For a large group such as snakes, broad-scale phylogenetic reconstructions largely rely on data integration. Increasing the compatibility of the data from different researches is thus important, which can be facilitated by standardization of the loci used in systematic analyses. In this study, we proposed a unified multilocus marker system for snake systematics by conflating 5 mitochondrial markers, 19 vertebrate-universal nuclear protein coding (NPC) markers and 72 snake-specific noncoding intron markers. This marker system is an addition to the large squamate conserved locus set (SqCL) for studies preferring a medium-scale data set. We applied this marker system to over 440 snake samples and constructed the currently most comprehensive systematic framework of the snakes in China. Robust snake phylogenetic relationships were recovered at both deep and shallow evolutionary depths, demonstrating the usefulness of this multilocus marker system. Discordance was revealed by a parallel comparison between the snake tree based on the multilocus marker system and that based on only the mitochondrial loci, highlighting the necessity of using multiple types of markers to better understand the snake evolutionary histories. The divergence times of different snake groups were estimated with the nuclear data set. Our comprehensive snake tree not only confirms many important nodes inferred in previous studies but also contributes new insights into many snake phylogenetic relationships. Suggestions are made for the current Chinese snake taxonomy.

Schizandrin attenuates lung lesions induced by Avian pathogenic Escherichia coli in chickens.

Avian pathogenic Escherichia coli (APEC) can cause serious pathological changes and inflammation in chickens. Schizandrin has anti-inflammatory activity and can prevent damage to various tissues and organs. The purpose of this study was to investigate the protective effect of schizandrin on APEC-induced lung lesions in chickens and explore the potential mechanism of schizandrin protection. The schizandrin (50, 100, and 200 mg/kg) was intragastrically administered for 3 days. APEC was administered using intraperitoneal (i.p.) injection to induce lung lesions. Then, chickens were sacrificed by CO2 inhalation 24 h later and the lung tissues were collected for examining histopathological changes, wet/dry (W/D) ratio, myeloperoxidase (MPO) activity, malondialdehyde (MDA), levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6, and IL-8 and activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. Our findings showed that schizandrin markedly inhibited pathological changes, pulmonary edema, MPO activity and MDA content. Moreover, schizandrin markedly reduced the levels of TNF-α, IL-1ß, IL-6 and IL-8 in lung tissue. Importantly, the mechanism responsible for these effects was attributed to the inhibitory effect of schizandrin on NF-κB and MAPK signaling activation. In conclusion, our findings reveal that schizandrin displays anti-oxidant and anti-inflammatory activity against APEC-induced lung lesions in chickens, paving the way for rational use of schizandrin as a protective agent against lung-related inflammatory disease.

Prevalence and genotype distribution of HPV and cervical pathological results in Sichuan Province, China: a three years surveys prior to mass HPV vaccination.

BACKGROUND: HPV persistent infection is a strong carcinogenic factor that can induce cervical cancer. Investigation of HPV epidemiology and genotype distribution is of great meaning for the development of cervical cancer prevention and control strategies. METHODS: By using PCR-based hybridization gene chip assay, HPV genotype was detected from 14,185 women that came from HEC (Health Examination Center) or OGOC (Obstetrics and Gynecology Outpatient Clinics) between 2015 and 2017 in Sichuan area. The epidemiology and genotype distribution as well as the relationship between HPV infection and histology/cytology abnormalities were analyzed. RESULTS: The positivity rate of HPV was 23.84%. The HPV-positive rate of OGOC group (37.62%) was significantly higher than that of HEC group (15.29%), p < 0.05. The prevalence of HPV reached peak at age 41-50 (5.86%) in HEC group, but at age 21-30 (14.74%) in OGOC group. Of all the HPV positive women, single genotype infection was the most common form in both HEC and OGOC group (62.06% in total screening population, 74.36% in HEC group and 54.01% in OGOC group). Three most prevalent HPV types were HPV-52 (5.02%), 58 (3.61%), and 16 (3.24%) in total screening population. Of all the HPV positive women, the top three types were HPV-52 (20.93%), CP8304 (15.32%), and 58 (14.42%) in HEC group, while were HPV-52 (21.14%), 16 (16.34%), and 58 (15.61%) in OGOC group. HPV 52/16/58 accounted for 41.84% of cytology and 56.52% of histological abnormalities. CONCLUSIONS: Women in Sichuan area were facing the great threat of HPV infection, especially the women aged between 21 ~ 30 or 41-50 years old. The priority HPV types were HPV 52, 58, and 16 in OGOC group, while were HPV 52, CP8304, and 58 in HEC group. HPV 52/16/58 accounted for the majority of cytology and histological abnormalities. Our analysis was found to be valuable for providing a scientific basis for the prevention and control strategies of cervical cancer in Sichuan area.

First Report of Round Leaf Spot on Sophora tonkinensis Caused by Didymella glomerata in China.

The root of Sophora tonkinensis Gagnep is an important medicinal material in China. An unknown foliar disease, first observed In July 2018, occurred over 240 ha of S. tonkinensis (totally cultured 600 ha) in Guangxi, China, in December 2019. The initial symptoms on leaf were seen as small, tawny spots (0.5 to 1.5 mm in diam.). As the disease progressed, the lesions enlarged into grayish and dark brown concentric rings (5 to 10.0 mm in diam.) resulting in black protuberances in the center of the spots. Severe infections would adversely affect plant growth, and cut the production by 30-40%. Symptomatic leaves were sampled and the surface was sterilized with 75% ethanol for 30 s and then soaked in 0.1% HgCl2 for 2 min. After three washes with sterile distilled water, the samples were dried, placed aseptically onto potato dextrose agar (PDA) plates, and incubated at 28°C. Three days later, the isolates were placed on new PDA medium for subsequent purification and sporulation. The fungus, SDG-1, was recovered from 85% of the total 40 isolates. Its colonies were whitish initially and then became olive green 7 days after incubation at 28 °C. The pycnidias were globose to subglobose, initially brown and darken at maturity, 85 to 300 × 70 to 280 µm in size. The conidia were colorless, single-celled, rounded to ellipsoidal and 3.5 to 6.6 × 1.5 to 3.8 µm. The chlamydospores were multicellular and brick trellised, usually forming branched or unbranched chains, light to dark brown in color and measured 28.5 to 44.5 × 8.2 to 16.5 µm. The morphological characteristics were consistent with Didymella glomerata. The rDNA-ITS, ß-tubulin and actin of strain SDG-1 were PCR amplified, and the DNA sequencing results were almost 100% identical to those of D. glomerata, respectively (GenBank database accession numbers MN 435377, MN447333 and MN447334, respectively). The multi-locus phylogenetic analysis was carried out with the obtained sequences, which revealed that the isolates clustered within D. glomerata with the similarity of 100% (Fig.3). Therefore, based on the morphology and phylogenetic tree, strain SDG-1 was identified as D. glomerata. For pathogenicity tests, the S. tonkinensis was cultured for two years, and the conidial suspension of SDG-1 (1 × 106 conidia /mL) was prepared by harvesting conidia from a 10-day-old culture on PDA. Conidia were sprayed onto the healthy leaves of S. tonkinensis for co-culture, while the control group was sprayed with sterile distilled water. Each experiment was performed three times. All plants were covered with plastic bags for 3 days in order to maintain high humidity and cultured in a greenhouse at 28 °C with a 12-h/12-h light/dark cycle. The symptoms appeared 7 days after the leaves were inoculated with spores, which were identical to those observed in the field. The pathogenic fungus was re-isolated from the infected leaves and identified as previously described. The control leaves remained symptomless during the pathogenicity tests. According to the previous literature, D. glomerata is a plant pathogenic fungus with a wide host range, which can damage up to 100 species of woody and herbaceous plants (Aghapour et al. 2009, Lahoz et al. 2007). To our knowledge, this is the first report of D. glomerata causing round leaf spot on S. tonkinensis in China.

[Role of mitochondrial damage in cadmium-induced cell apoptosis and DNA damage of hepatocytes].

OBJECTIVE: To investigate the role of mitochondrial damage mediated by reactive oxidative species(ROS) in cadmium-induced cell apoptosis and DNA damage of L02 hepatocytes, so as to provide experimental basis for the subsequent study and protection of people exposed to Cd. METHODS: The L02 hepatocytes were cultured in vitro treated with 0-90 µmol/L Cd for 24 h, and the methylthiazolyldiphenyltetrazoliumbromide assay was used to detect the cell viability. The colony formation assay, flow cytometry, comet assay, 2', 7'-dichlorofluorescein diacetate, MitoTracker Red CMXRos and 10-N-nonyl-acridine-orange, mitochondrial membrane potential detection kit(JC-1) and adenosine triphosphate(ATP) assay kits and Western Blot were used to investigate cell growth and proliferation, cell apoptosis, DNA damage, ROS levels, mitochondrial morphology, mitochondrial membrane potential, mitochondrial mass, ATP content and related proteins after the cells exposed to 0, 20, 40 µmol/L Cd for 24 h. The cells were pretreated with vitamin C before adding Cd exposure, and ROS levels, mitochondrial function, cell apoptosis, DNA damage and proteins were measured. RESULTS: The cell viability was significantly inhibited with the increase of Cd concentration and treatment time. The cells were treated with Cd for 24 h for further study according to the result of MTT assay. Compared with control group, the colony formation rate were 8. 23% and 6. 17% respectively in 20 and 40 µmol/L Cd treatment and the apoptosis rate were 15. 85% and 26. 26%, respectively. We also found that the B cell lymphoma/leukemia(Bcl-2) gene protein was significantly reduced, while the levels of Bcl-2 associated X protein(Bax) and cleaved cysteine aspastic acid-specific protease 3(cleaved-caspase-3) were increased in a dose-dependent manner. Cd treatment also induced DNA damage and accumulation of intracellular ROS, accompanied by a mitochondrial morphological change, significant decrease in Δψm, mitochondrial mass, ATP content, mitochondrial cytochrome C(cyt c) and an increase in cytoplasmic cyt c expression(P<0. 05). In addition, pretreatment with antioxidant vitamin C not only significantly increased cyt c, mitochondrial mass, ATP content and mitochondrial cyt c, but also reduced the expression of cytoplasmic cyt c(P<0. 05), cell apoptosis and DNA damage induced by Cd. CONCLUSION: Cd exposure could induce ROS accumulation in L02 hepatocytes, which can lead to mitochondrial damage, and ultimately lead to cell apoptosis and DNA damage.

Characterizing 3D inflorescence architecture in grapevine using X-ray imaging and advanced morphometrics: implications for understanding cluster density.

Inflorescence architecture provides the scaffold on which flowers and fruits develop, and consequently is a primary trait under investigation in many crop systems. Yet the challenge remains to analyse these complex 3D branching structures with appropriate tools. High information content datasets are required to represent the actual structure and facilitate full analysis of both the geometric and the topological features relevant to phenotypic variation in order to clarify evolutionary and developmental inflorescence patterns. We combined advanced imaging (X-ray tomography) and computational approaches (topological and geometric data analysis and structural simulations) to comprehensively characterize grapevine inflorescence architecture (the rachis and all branches without berries) among 10 wild Vitis species. Clustering and correlation analyses revealed unexpected relationships, for example pedicel branch angles were largely independent of other traits. We identified multivariate traits that typified species, which allowed us to classify species with 78.3% accuracy, versus 10% by chance. Twelve traits had strong signals across phylogenetic clades, providing insight into the evolution of inflorescence architecture. We provide an advanced framework to quantify 3D inflorescence and other branched plant structures that can be used to tease apart subtle, heritable features for a better understanding of genetic and environmental effects on plant phenotypes.