Splitting one species into 22: an unusual tripling of molecular, morphological, and geographical differentiation in the fern family Didymochlaenaceae (Polypodiales).

The pantropical fern genus Didymochlaena (Didymochlaenaceae) has long been considered to contain one species only. Recent studies have resolved this genus/family as either sister to the rest of eupolypods I or as the second branching lineage of eupolypods I, and have shown that this genus is not monospecific, but the exact species diversity is unknown. In this study, a new phylogeny is reconstructed based on an expanded taxon sampling and six molecular markers. Our major results include: (i) Didymochlaena is moderately or weakly supported as sister to the rest of eupolypods I, highlighting the difficulty in resolving the relationships of this important fern lineage in the polypods; (ii) species in Didymochlaena are resolved into a New World clade and an Old World clade, and the latter further into an African clade and an Asian-Pacific clade; (iii) an unusual tripling of molecular, morphological and geographical differentiation in Didymochlaena is detected, suggesting single vicariance or dispersal events in individual regions and no evidence for reversals at all, followed by allopatric speciation at more or less homogeneous rates; (iv) evolution of 18 morphological characters is inferred and two morphological synapomorphies defining the family are recognized-the elliptical sori and fewer than 10 sori per pinnule, the latter never having been suggested before; (v) based on morphological and molecular variation, 22 species in the genus are recognized contrasting with earlier estimates of between one and a few; and (vi) our biogeographical analysis suggests an origin for Didymochlaena in the latest Jurassic-earliest Cretaceous and the initial diversification of the extant lineages in the Miocene-all but one species diverged from their sisters within the last 27 Myr, in most cases associated with allopatric speciation owing to geologic and climatic events, or dispersal.

[Adionuclide therapy for metastatic castration-resistant prostate cancer: New advances].

Prostate cancer (PCa) is one of the most common tumors of the male urogenital system, ranking the second among male malignancies worldwide. Age is a major risk factor for PCa, and population aging leads to an increasing incidence of the malignancy. Androgen-deprivation therapy (ADT) is currently the first-line treatment of PCa, but with the advance of the tumor, many of the patients become resistant to ADT and develop castration-resistant prostate cancer (CRPC), which marks a transition of PCa to a hormone-refractory state associated with a poor prognosis. Metastatic CRPC (mCRPC) is the terminal stage of the disease and a leading cause of death. Despite many new advances in the treatment of mCRPC, its prognosis remains poor. For patients with mCRPC, classical treatment options with chemotherapy and androgen-blocking agents have limited survival benefit, and radionuclide therapy has become a novel therapeutic option for mCRPC. This paper focuses on the development of the radionuclide therapy for mCRPC in recent years.

[Cuproptosis-related lncRNAs to predict biochemical recurrence of prostate cancer].

OBJECTIVE: To construct a cuproptosis-related lncRNA model and obtain some new ideas and methods for predicting the biochemical recurrence (BCR) of PCa. METHODS: We identified cuproptosis-related lncRNAs from the gene expression data, mutation load data and clinical data on PCa patients in the Cancer Genome Atlas (TCGA) database and divided the patients into a training group and a verification group. We constructed a prognostic risk scoring model based on the cuproptosis -related lncRNAs, verified the validity of the model by BCR-free survival analysis, logistic regression analysis and independent prognosis analysis, and visualized the results using ROC curve analysis, Kaplan-Meier survival curves and the correlation heat map. We performed differential analysis and survival analysis of the tumor mutation burden (TMB), and assessed the value of the model and TMB in predicting the BCR of PCa. RESULTS: A prognostic risk scoring model was successfully constructed based on the 6 cuproptosis -related lncRNAs identified from the PCa cases in the training group, which were divided into a high- and a low-risk groups according to the median value. The incidence of BCR rose with the increase of the risk score, and the BCR-free time was significantly shorter in the high-risk group (P < 0.05). The model also exhibited a high differentiation value in different age groups (P < 0.05), which was shown to be a reliable and independent prognostic indicator for predicting the BCR of PCa, even more valuable than other clinicopathological indicators. TMB was differentially expressed in the high- and low-risk groups (P < 0.01) and significantly correlated with BCR. The highest rate of BCR-free survival was found in the patients with low risk scores and low TMB (P < 0.01). CONCLUSION: A cuproptosis -related lncRNA model was successfully constructed, which can accurately predict the risk of BCR in PCa patients. The higher the prognostic risk score, the greater the possibility of BCR. TMB is high in patients with a high risk, and the TMB level has certain suggestive significance for BCR.

[Influence factors of erectile dysfunction in patients with localized prostate cancer after radical surgery].

OBJECTIVE: To analyze the influential factors of erectile dysfunction (ED) in patients with localized prostate cancer (LPC) after radical surgery. METHODS: The clinical data of 150 male patients diagnosed with LPC and normal erectile function (EF) before surgery admitted to the Department of Urology of the Eastern Theatre General Hospital from January 2021 to January 2023 were retrospectively analyzed. The EF status of the patients 6 months after surgery was assessed using the International Erectile Function Index -5(IIEF-5). Age, Gleason score, PSA level, TNM stage, preoperative International prostatic symptom score (IPSS), preoperative prostate volume, smoking index, alcohol consumption index, educational level, comorbidities, operation mode, and psychosexual state were used as influencing factors to analyze their effects on postoperative ED. RESULTS: Of the 150 patients, 88 had ED and 62 had normal EF. Univariate analysis showed that age, preoperative IPSS, preoperative prostate volume, comorbidities and sexual and psychological status were significantly correlated with postoperative ED. Further multivariate logistic regression analysis showed that age, preoperative prostate volume, comorbidities and sexual and psychological status were independent factors influencing the occurrence of ED after RP in LPC patients. CONCLUSION: The recovery of sexual function of patients with localized prostate cancer after radical surgery is generally poor, and the incidence of ED is high. Its independent influencing factors include age, preoperative prostate volume, comorbidities and sexual psychological state, etc. Correct intervention of different influencing factors is required in clinical work. In order to provide a better diagnosis and treatment scheme for LPC patients undergoing radical treatment, reduce the incidence of postoperative ED and improve the quality of life of patients after surgery.

When immunotherapy meets liver transplantation for hepatocellular carcinoma: A bumpy but promising road.

Liver transplantation (LT) is a highly curative therapy for patients with hepatocellular carcinoma (HCC). However, due to the shortage of donor livers and rapid progression of HCC, a majority of patients are dropped out from the waitlist. Recently, immunotherapy has shown great promise in the treatment of advanced HCC. However, the use of immunotherapy is limited in LT mainly due to the potentially increasing risk of graft rejection. One of the main challenges for researchers is the protection of donor graft from an immunotherapy-boosted immune response mounted by the host. Besides, the safety, availability, and costs of immunotherapy are other challenges that need to be addressed. Here, we reviewed the literature involving patients who received immunotherapy prior to transplant to avoid waitlist dropouts and following transplantation to prevent the progression of tumor recurrence and metastasis. Statistically, the incidence of rejection was 25.0% pre-transplant and 18.5% post-transplant. Based on the review of these clinical studies, we can conclude that conducting clinical trials on the safety and efficacy of currently available immunotherapy drugs and identifying novel immunotherapy targets through extensive research may be promising for patients who do not meet the selection criteria for LT and who experience post-transplant recurrence. To date, the clinical experience on the use of immunotherapy before or after LT comes from individual case studies. Although some of the reported results are promising, they are not sufficient to support the standardized use of immunotherapy in clinical practice.

Coexistence of virulence and ß-lactamase genes in avian pathogenic Escherichia coli.

Emergence of multidrug resistance in E. coli and advent of newer strains is becoming serious concern which requires keen observations. This study was designed to find the ciprofloxacin resistant E. coli isolates co-existed with multi-drug resistance along with ß-lactamase production from poultry source, and finally the genome sequencing of these strains to explore genetic variations. Study constituted on isolation of n = 225 E. coli from broiler farms of central China which were further subjected to identification of resistance against ciprofloxacin followed by antibiogram of n = 26 antibiotics and identification of ß-lactamase production. Whole genome resequencing was performed using Illumina HiSeq 4000 system. PCR results revealed predominant ß-lactamase genes i.e.CTX-M, CTX-M-1, CTX-M3, TEM-1 and OXA. Furthermore, the MDR isolates were containing most of the tested virulence genes. The most prevalent virulence genes were pap-C, fim-C, fim-H, iuc-D, irp-2, tra-T, iro-N and iut-A. The single nucleotide polymorphisms (SNPs) loci mentioned in this data give valuable genetic markers to growing high-throughput techniques for fine-determination of genotyping of MDR and virulent isolates. Characterization of SNPs on functional basis shed new bits of knowledge on the evolution, disease transmission and pathogenesis of MDR E. coli isolates. In conclusion, these findings provide evidence that most of poultry E. coli are MDR, ß-lactamase producers, and virulent which could be a zoonotic threat to the humans. The whole genome resequencing data provide higher resolution of resistance and virulence characteristics in E. coli which can further be used for the development of prevention and treatment strategies.

The reference ranges and characteristics of lymphocyte parameters and the correlation between lymphocyte parameters and routine health indicators in adults from China.

BACKGROUND: Assessment of immune function is of key importance in recognition of disease or healthy status, which still faces challenge in clinical practice. We conducted a 10-center study to investigate lymphocyte parameters including the number, phenotype and IFN-γ-producing ability, and routine laboratory indicators by using the standard method. RESULTS: Although the heterogeneity of lymphocyte parameters was widely found, we have established the normal ranges of these parameters by using pooled data which showed no significant difference among centers. Cluster analysis of 35 parameters found 3 interesting clusters which represented different immunological status. Cluster 1 (parameters: IFN-γ+CD4+ T cell percentage and IFN-γ+CD8+ T cell percentage) represented current lymphocyte function, which was associated with indicators such as body mass index and red blood cell; Cluster 2 (parameters: NK cell number and CD45RA+CD4+ T cell percentage) represented potential of lymphocytes, which was associated with indicators such as albumin and high-density lipoprotein. Cluster 3 (parameters: HLA-DR+CD8+ T cell percentage) represented inflammatory status, which was associated with indicators such as low-density lipoprotein, globulin and age. Correlation analysis found that nutritional indicator albumin is significantly positively correlated with lymphocyte potential. Triglyceride and body mass index were positively correlated with current lymphocyte function rather than lymphocyte potential. The loss of CD8+ T cells was extremely pronounced with increasing age and was one of the most important factors to cause immunosenescence, which may be associated with increased glucose. CONCLUSIONS: We have established the normal ranges of lymphocyte parameters in different areas. This study elucidates the key indicators used to reflect the current function or potential of lymphocytes, which may provide a valuable clue for how to keep immunity healthy.

Clinical Breakpoint of Apramycin to Swine Salmonella and Its Effect on Ileum Flora.

The purpose of this study was to establish the clinical breakpoint (CBP) of apramycin (APR) against Salmonella in swine and evaluate its effect on intestinal microbiota. The CBP was established based on three cutoff values of wild-type cutoff value (COWT), pharmacokinetic-pharmadynamic (PK/PD) cutoff value (COPD) and clinical cutoff value (COCL). The effect of the optimized dose regimen based on ex vivo PK/PD study. The evolution of the ileum flora was determined by the 16rRNA gene sequencing and bioinformatics. This study firstly established the COWT, COPD in ileum, and COCL of APR against swine Salmonella, the value of these cutoffs were 32 µg/mL, 32 µg/mL and 8 µg/mL, respectively. According to the guiding principle of the Clinical Laboratory Standards Institute (CLSI), the final CBP in ileum was 32 µg/mL. Our results revealed the main evolution route in the composition of ileum microbiota of diarrheic piglets treated by APR. The change of the abundances of Bacteroidetes and Euryarchaeota was the most obvious during the evolution process. Methanobrevibacter, Prevotella, S24-7 and Ruminococcaceae were obtained as the highest abundance genus. The abundance of Methanobrevibacter increased significantly when APR treatment carried and decreased in cure and withdrawal period groups. The abundance of Prevotella in the tested groups was significantly lower than that in the healthy group. A decreased of abundance in S24-7 was observed after Salmonella infection and increased slightly after cure. Ruminococcaceae increased significantly after Salmonella infection and decreased significantly after APR treatment. In addition, the genera of Methanobrevibacter and Prevotella were defined as the key node. Valine, leucine and isoleucine biosynthesis, D-Glutamine and D-glutamate metabolism, D-Alanine metabolism, Peptidoglycan and amino acids biosynthesis were the top five Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways in the ileum microbiota of piglets during the Salmonella infection and APR treatment process. Our study extended the understanding of dynamic shift of gut microbes during diarrheic piglets treated by APR.

[Erectile function of renal failure patients after hemodialysis or kidney transplantation: A comparative study].

OBJECTIVE: To study the changes in the erectile function of the male patients with renal failure after hemodialysis (HD) or kidney transplantation (KT) and explore the causes of these changes. METHODS: From January 2015 to January 2021, 160 male patients with renal failure complaining of ED underwent HD (n = 80) or KT (n = 80) in the General Hospital of Eastern Theater Command. The patients were aged 25－45 (31.7 ± 4.8) years, 32 ± 4.5 years in the HD group and 31.4 ± 5.1 years in the KT group. We recorded the levels of serum T, E2, FSH and LH and the scores on IIEF-5, Self-Rating Anxiety Scale (SAS) and Self-Rating Depression Scale (SDS) of the patients, and compared them between the two groups. RESULTS: Compared with the patients in the HD group, those in the KT group showed a significantly higher T level (ï¼»7.45 ± 3.54ï¼½ vs ï¼»17.75 ± 7.32ï¼½ nmol/L, P < 0.01) and a lower E2 level (ï¼»151.37 ± 20.89ï¼½ vs ï¼»94.17 ± 40.79ï¼½ pmol/L, P < 0.01), but no statistically significant difference from the former group in the levels of FSH (ï¼»8.12 ± 5.12ï¼½ vs ï¼»8.97 ± 2.36ï¼½ IU/L, P > 0.05) and LH (ï¼»5.16 ± 3.87ï¼½ vs ï¼»4.69 ± 2.18ï¼½ IU/L, P > 0.05). There were fewer cases of severe ED in the KT than in the HD group (3.75% vs 16.25%, P < 0.05). Different degrees of anxiety and depression were observed in both groups, with fewer severe cases of anxiety (6.25% vs 30.00%, P < 0.05) and depression (6.25% vs 31.25%, P < 0.05) and more mild cases of anxiety (68.75% vs 47.50%, P < 0.05) and depression (70.00% vs 48.75%, P < 0.05) in the KT than in the HD group, but no statistically significant difference in the incidence of moderate anxiety (25.00% vs 22.50%, P > 0.05) and depression (23.75% vs 20.00%, P > 0.05) between the KT and HD groups. CONCLUSION: For male patients with renal failure, kidney transplantation can evidently improve erectile function, while hemodialysis has a poorer effect. The altered hormone levels, anxiety and depression of the patients are important causes of the changes in their erectile function.

AMPK activation by ozone therapy inhibits tissue factor-triggered intestinal ischemia and ameliorates chemotherapeutic enteritis.

Chemotherapeutic enteritis is a major dose-limiting adverse reaction to chemotherapy, with few effective drugs in clinic. Intestinal ischemic injury plays prominent role in chemotherapeutic enteritis clinically. However, mechanism is not clear. In this article, irinotecan (CPT-11) was used to establish chemotherapeutic enteritis mice model. Western blotting, gelatin zymography, immunohistochemistry (IHC), Laser Doppler flowmetry (LDF) were used to detect the pathogenesis of ischemia-hypoxia injury. CPT-11 increased levels of tissue factor (TF) both in the blood and in intestines, and decreased the intestinal blood flow in mice. Interestingly, the elevation of TF in the blood displayed "double-peak," which was consistent with the intestinal mucosal "double-strike" injury trend. Intestinal microthrombus and mixed thrombus formation were detectable in chemotherapeutic enteritis. Furthermore, ozone therapy relieved chemotherapeutic enteritis in mice. Ozone inhibited TF expression induced by CPT-11 via activating AMPK/SOCS3, and effectively ameliorated the intestinal mucosal injury in mice. Moreover, ozone autotransfusion therapy effectively attenuated chemotherapeutic enteritis and the blood hypercoagulability in patients. For the first time, we proposed that TF-induced thrombotic intestinal ischemic injury is a core trigger pathological mechanism of chemotherapeutic enteritis, and provided a new treatment strategy, ozone therapy, to suppress TF expression and treat chemotherapeutic enteritis.

The Evolution of Fluoroquinolone Resistance in Salmonella under Exposure to Sub-Inhibitory Concentration of Enrofloxacin.

The evolution of resistance in Salmonella to fluoroquinolones (FQs) under a broad range of sub-inhibitory concentrations (sub-MICs) has not been systematically studied. This study investigated the mechanism of resistance development in Salmonella enterica serovar Enteritidis (S. Enteritidis) under sub-MICs of 1/128×MIC to 1/2×MIC of enrofloxacin (ENR), a widely used veterinary FQ. It was shown that the resistance rate and resistance level of S. Enteritidis varied with the increase in ENR concentration and duration of selection. qRT-PCR results demonstrated that the expression of outer membrane porin (OMP) genes, ompC, ompD and ompF, were down-regulated first to rapidly adapt and develop the resistance of 4×MIC, and as the resistance level increased (≥8×MIC), the up-regulated expression of efflux pump genes, acrB, emrB amd mdfA, along with mutations in quinolone resistance-determining region (QRDR) gradually played a decisive role. Cytohubba analysis based on transcriptomic profiles demonstrated that purB, purC, purD, purF, purH, purK, purL, purM, purN and purT were the hub genes for the FQs resistance. The 'de novo' IMP biosynthetic process, purine ribonucleoside monophosphate biosynthetic process and purine ribonucleotide biosynthetic process were the top three biological processes screened by MCODE. This study first described the dynamics of FQ resistance evolution in Salmonella under a long-term selection of sub-MICs of ENR in vitro. In addition, this work offers greater insight into the transcriptome changes of S. Enteritidis under the selection of ENR and provides a framework for FQs resistance of Salmonella for further studies.

The Effects of Natural Products and Environmental Conditions on Antimicrobial Resistance.

Due to the extensive application of antibiotics in medical and farming practices, the continued diversification and development of antimicrobial resistance (AMR) has attracted serious public concern. With the emergence of AMR and the failure to treat bacterial infections, it has led to an increased interest in searching for novel antibacterial substances such as natural antimicrobial substances, including microbial volatile compounds (MVCs), plant-derived compounds, and antimicrobial peptides. However, increasing observations have revealed that AMR is associated not only with the use of antibacterial substances but also with tolerance to heavy metals existing in nature and being used in agriculture practice. Additionally, bacteria respond to environmental stresses, e.g., nutrients, oxidative stress, envelope stress, by employing various adaptive strategies that contribute to the development of AMR and the survival of bacteria. Therefore, we need to elucidate thoroughly the factors and conditions affecting AMR to take comprehensive measures to control the development of AMR.

PGE2 ameliorated viral myocarditis development and promoted IL-10-producing regulatory B cell expansion via MAPKs/AKT-AP1 axis or AhR signaling.

B10 cells, a specific subset of regulatory B cells, are capable of regulating immune response and restricting inflammation and autoimmune disease progression by producing IL-10. B10 cells frequently change significantly during inflammation and autoimmunity. However, how B10 cell populations change in viral myocarditis (VMC) remains unclear. Therefore, this work was conducted to clarify the changes in B10 cells and their potential mechanisms. Our results showed that the B10 cell frequency significantly changed in the VMC model. Changes in prostaglandin E2 (PGE2) levels in VMC model hearts were consistent with B10 expansion. PGE2 induced B10 cell expansion via the MAPKs/AKT-AP1 axis or AhR signaling. Additionally, PGE2-pretreated B10 cells inhibited naïve CD4+ T cell differentiation into Th17 cells. In vivo, PGE2 treatment or adoptive B10 cell transfer significantly restricted VMC development. Our results provide sufficient evidence that PGE2-induced B10 cell expansion may become a promising therapeutic approach for VMC and acute inflammatory injury.

Inhibition of apoptosis signal-regulating kinase by paeoniflorin attenuates neuroinflammation and ameliorates neuropathic pain.

BACKGROUND: Neuropathic pain is a serious clinical problem that needs to be solved urgently. ASK1 is an upstream protein of p38 and JNK which plays important roles in neuroinflammation during the induction and maintenance of chronic pain. Therefore, inhibition of ASK1 may be a novel therapeutic approach for neuropathic pain. Here, we aim to investigate the effects of paeoniflorin on ASK1 and neuropathic pain. METHODS: The mechanical and thermal thresholds of rats were measured using the Von Frey test. Cell signaling was assayed using western blotting and immunohistochemistry. RESULTS: Chronic constrictive injury (CCI) surgery successfully decreased the mechanical and thermal thresholds of rats and decreased the phosphorylation of ASK1 in the rat spinal cord. ASK1 inhibitor NQDI1 attenuated neuropathic pain and decreased the expression of p-p38 and p-JNK. Paeoniflorin mimicked ASK1 inhibitor NQDI1 and inhibited ASK1 phosphorylation. Paeoniflorin decreased the expression of p-p38 and p-JNK, delayed the progress of neuropathic pain, and attenuated neuropathic pain. Paeoniflorin reduced the response of astrocytes and microglia to injury, decreased the expression of IL-1ß and TNF-α, and downregulated the expression of CGRP induced by CCI. CONCLUSIONS: Paeoniflorin is an effective drug for the treatment of neuropathic pain in rats via inhibiting the phosphorylation of ASK1, suggesting it may be effective in patients with neuropathic pain.

Down-regulation of lncRNA OGFRP1 induces autophagy and growth inhibition by AKT/mTOR signaling pathway in HCAECs.

Long noncoding RNAs (lncRNAs) have been found to play important roles in nearly all biological processes. However, the functions of the majority of LncRNAs are not fully clear. Here we evaluated the function of lncRNA OGFRP1, which has not been previously annotated, in human coronary artery endothelial cells (HCAECs). First, we knocked down lncRNA OGFRP1 in HCAECs by using siRNA transfection. qRT-PCR results indicated that siRNA1 and siRNA3 both had potent interference efficiencies. Next, by using CCK8 assay and clone formation assay, we found that siRNA3 transfection induced growth inhibition in HCAECs. Cell migration and invasion were also found to be inhibited in OGFRP1 silenced cells. Moreover, siRNA1 transfection further verified the inhibitory effects of lncRNA OGFRP1 knockdown on the proliferation, migration and invasion of HCAECs. Flow cytometry detection demonstrated that OGFRP1 knockdown induced cell cycle arrest and apoptosis. Western blot assay indicated that p70S6K and CyclinD1 were down-regulated by knockdown of OGFRP1. The intrinsic apoptosis pathway was activated in lncRNA OGFRP1 silenced cells, including increased Bax and Active-caspase 3 and decreased Bcl2. The expression of autophagy markers LC3 and Beclin1 was increased and p62 decreased, all of which indicated that cell autophagy was promoted by down-regulation of lncRNA OGFRP1. Mechanistic studies showed that lncRNA OGFRP1 inhibited the AKT/mTOR signaling pathway, including increasing phosphorylation level of AKT, mTOR and GSK3ß. In conclusion, we find that down-regulation of lncRNA induces autophagy and inhibited the proliferation, migration and invasion by AKT/mTOR signaling pathway in HCAECs.

Outdoor Scene Understanding Based on Multi-Scale PBA Image Features and Point Cloud Features.

Outdoor scene understanding based on the results of point cloud classification plays an important role in mobile robots and autonomous vehicles equipped with a light detection and ranging (LiDAR) system. In this paper, a novel model named Panoramic Bearing Angle (PBA) images is proposed which is generated from 3D point clouds. In a PBA model, laser point clouds are projected onto the spherical surface to establish the correspondence relationship between the laser ranging point and the image pixels, and then we use the relative location relationship of the laser point in the 3D space to calculate the gray value of the corresponding pixel. To extract robust features from 3D laser point clouds, both image pyramid model and point cloud pyramid model are utilized to extract multiple-scale features from PBA images and original point clouds, respectively. A Random Forest classifier is used to accomplish feature screening on extracted high-dimensional features to obtain the initial classification results. Moreover, reclassification is carried out to correct the misclassification points by remapping the classification results into the PBA images and using superpixel segmentation, which makes full use of the contextual information between laser points. Within each superpixel block, the reclassification is carried out again based on the results of the initial classification results, so as to correct some misclassification points and improve the classification accuracy. Two datasets published by ETH Zurich and MINES ParisTech are used to test the classification performance, and the results show the precision and recall rate of the proposed algorithms.

Biological function of hpsh4590 localized in the plasticity zone of Helicobacter pylori.

The aim of this study was to determine the biological function of hpsh4590 in Helicobacter pylori. After Hpsh4590 was expressed using a prokaryotic expression system, the cytotoxic effects and IL-8 production of Hpsh4590 were analyzed by co-culturing with GES-1 cells. Meanwhile, the antibody of rHpsh4590, produced by immunizing rabbit, was used for localization and protein interaction studies. Hpsh4590 fusion protein was expressed successfully in Escherichia coli Rosetta (DE3), and the polyclonal antibody was produced at high titers. The MTT assay showed that the inhibition ratio of GES-1 cells cultured with 0.1 µg/mL rHpsh4590 (3.02% ± 0.02%) was significantly lower than that of 20 µg/mL rHpsh4590 (57.57% ± 0.03%, p < 0.01), while DAPI staining showed the cytotoxic effects of rHpsh4590 for GES-1 cells. The up-regulation of cleaved caspase-3 and cleaved PARP was observed after GES-1 cells co-cultured with rHpsh4590 by Western blot. Co-culturing of GES-1 cells with rHpsh0459 (20 µg/mL) led to significant production of IL-8 at 12 h(1097.74 ± 212.37 pg/mL) and 24 h (1379.55 ± 209.58 pg/mL) then at 6 h(134.68 ± 14.64 pg/mL, p < 0.01). These observations suggest that the cytotoxicity of Hpsh4590 occurred in a concentration dependent manner, which is related with IL-8 secretion from gastric mucosal epithelial cells. Hpsh4590 was found localized in the membrane and the periplasm of H. pylori, interacted with zinc finger protein and methionine ABC transporter ATP-binding protein, and potentially regulates DNA uptake or transfer.

The complete chloroplast genome sequence of Isoetes baodongii (Isoetaceae).

Isoetes baodongii is a diploid species of Isoetaceae distributed in low altitude area, its megaspore ornamentation is similar to tetraploid species I. sinensis. We collected leaf material of I. baodongii and sequenced it for low depth whole genome sequence, then, a complete chloroplast genome of I. baodongii was assembled and annotated. This chloroplast genome has a circular structure of 145,494 bp in length with a GC content of 38.0%, comprising a large single copy (LSC) region of 91,860 bp, a pair of inverted repeat (IR) regions of 13,207 bp each, and a small single copy (SSC) region of 27,220 bp. 136 genes were annotated, including 84 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. A maximum likelihood phylogeny tree was reconstructed after the sequences alignment, the result showed that I. baodongii formed a sister clade to the one clustered by I. sinensis, I. taiwanensis and I. orientalis. Although the chloroplast genome structure of I. baodongii is extremely similar to other species distributed in China, a well-supported phylogenetic relationship was reconstructed here, these results may provide new messages for further studies on phylogeny and evolution of vascular plant on the earth.

A novel variant in the FLCN gene in a Chinese family with Birt-Hogg-Dubé syndrome.

BACKGROUND: This study aimed to identify disease-causing variants within a Chinese family affected by Birt-Hogg-Dubé syndrome (BHDS), which arises from an autosomal dominant inheritance pattern attributed to variants in the folliculin (FLCN) gene, recognized as a tumor suppressor gene. METHODS: A Chinese proband diagnosed with BHDS due to renal tumors underwent next-generation sequencing (NGS), revealing a novel variant in the FLCN gene. Sanger sequencing was subsequently performed on blood samples obtained from family members to confirm the presence of this variant. RESULTS: A novel germline frameshift variant (NM_144997.5:c.977dup) was identified in five individuals among the screened family members, marking the first report of this variant. Additionally, a somatic frameshift variant (NM_144997.5:c.1252del) was detected in the renal tumors of the proband. No variant was detected in unaffected family members. CONCLUSIONS: A novel heterozygous variant was identified in exon 9 of the FLCN gene, which broadens the spectrum of FLCN variants. We recommend that molecular analysis of the FLCN gene be performed in patients with suspected BHDS and their families.

Genus Shewanella: A Potential Intestinal Colonizer Associated With Post-Operative Surgical Site Infections in Coastal Regions.

Background: This study aims to elucidate the clinical characteristics of Shewanella-related surgical site infections (SSIs) and assess the risk of mortality in patients by establishing a predictive model. Patients and Methods: A retrospective analysis of medical history and laboratory data of Shewanella-related SSI patients over the past decade was conducted via the electronic medical record (EMR) system. A predictive model for mortality risk in Shewanella-related SSI patients was established using plasma interleukin-6 (IL-6) levels combined with the Howell-PIRO scoring system. Results: Over the past 10 years, 45 strains of Shewanella were isolated from specimens such as bile, drainage fluid, and whole blood in patients with digestive tract SSIs. Among them, 21 of 45 (46.67%) patients underwent malignant tumor resection of the digestive system, 14 of 45 (31.11%) underwent endoscopic retrograde cholangiopancreatography (ERCP) common bile duct exploration or the stone removal, and seven of 45 (15.56%) were trauma repair patients with fractures and abdominal injuries. Among the 45 Shewanella-related SSI patients, 10 died within 30 days of infection, six cases involved infections with more than two other types of bacteria. The combined use of IL-6 and Howell-PIRO scores for mortality risk assessment yielded an receiver operating characteristic (ROC) curve with an area under the curve (AUC) of 0.9350, a positive predictive value of 92.71%, a negative predictive value of 94.58%, a diagnostic sensitivity of 95.35%, and a diagnostic specificity of 92.14%-all higher than the model using IL-6 or Howell-PIRO scores alone. Conclusions: We found that residents in coastal areas faced an increased risk of Shewanella-related SSI. Moreover, the higher the number of concurrent microbial infections occurring alongside Shewanella-related SSI, the greater the mortality rate among patients. The combined application of plasma IL-6 levels and the Howell-PIRO scoring system is beneficial for assessing patient mortality risk and guiding timely and proactive clinical interventions.