Antifungal bioactivity of Sarcococca hookeriana var. digyna Franch. against fluconazole-resistant Candida albicans in vitro and in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Sarcococca hookeriana var. digyna Franch. has been widely utilized in folk medicine by the Miao people in the southwestern region of China for treating skin sores which may be associated with microbial infection. AIM OF THE STUDY: To investigate the antifungal bioactivity of S. hookeriana var. digyna against fluconazole-resistant Candida albicans in vitro and in vivo, as well as its underlying mechanism and the key bioactive component. MATERIALS AND METHODS: The antifungal bioactivity of 80% ethanol extract of S. hookeriana var. digyna (SHE80) was investigated in vitro using the broth microdilution method, time-growth curve, and time-kill assay. Its key functional component and antifungal mechanism were explored with combined approaches including UPLC-Q-TOF-MS, network pharmacology and metabolomics. The antifungal pathway was further supported via microscopic observation of fungal cell morphology and examination of its effects on fungal biofilm and cell membranes using fluorescent staining reagents. In vivo assessment of antifungal bioactivity was conducted using a mouse model infected with C. albicans on the skin. RESULTS: S. hookeriana var. digyna suppressed fluconazole-resistant C. albicans efficiently (MIC = 16 µg/mL, MFC = 64 µg/mL). It removed fungal biofilm, increased cell membrane permeability, induced protein leakage, reduced membrane fluidity, disrupted mitochondrial membrane potential, induced the release of reactive oxygen species, promoted cell apoptosis, and inhibited the transformation of fungi from the yeast state to the hyphal state significantly. In terms of mechanism, it affected sphingolipid metabolism and signaling pathway. Moreover, the predicted bioactive component, sarcovagine D, was supported by antifungal bioactivity evaluation in vitro (MIC = 4 µg/mL, MFC = 16 µg/mL). Furthermore, S. hookeriana var. digyna promoted wound healing, reduced the number of colony-forming units, and reduced inflammation effectively in vivo. CONCLUSIONS: The traditional use of S. hookeriana var. digyna for fungal skin infections was supported by antifungal bioactivity investigated in vitro and in vivo. Its mechanism and bioactive component were predicted and confirmed by experiments, which also provided a new antifungal agent for future research.

A Case of Rectal Endometriosis Misdiagnosed as Rectal Malignancy on Three Colonoscopies and Biopsies Sharing a Combined Literature Review.

Background: Endometriosis involves the intestine, and its clinical manifestations are nonspecific and lack of etiological manifestations. The diagnosis is difficult, which often leads to misdiagnosis. We report a case of endometriosis which was misdiagnosed as intestinal malignant tumor after colonoscopy and three biopsies. Case Presentation: We reported a 42-year-old woman who went to see a doctor because of anal distension. She was examined by three gastrointestinal endoscopists at different levels in different hospitals and underwent biopsy at the same time. Combined with clinical manifestations, imaging examination, endoscopic examination and pathological examination, she was misdiagnosed as intestinal malignant tumor, and partial intestinal resection was performed according to the surgical principle of malignant tumor. Conclusion: Although there are advanced gastrointestinal endoscopy and imaging techniques, intestinal endometriosis is still easy to be misdiagnosed. As our case report shows, after three colonoscopy and biopsy, it is still misdiagnosed as intestinal malignant tumor. Further research is needed to improve the ability of preoperative diagnosis, which deserves the attention of gastroenterologists and obstetricians and gynecologists.

Development and validation of a novel anoikis-related gene signature for predicting prognosis in ovarian cancer.

Anoikis plays a critical role in variable cancer types. However, studies that focus on the prognostic values of anoikis-related genes (ANRGs) in OV are scarce. Cohorts with transcriptome data and corresponding clinicopathologic data of OV patients were collected and consolidated from public databases. Multiple bioinformatics approaches were used to screen key genes from 446 anoikis-related genes, including Cox regression analysis, random survival forest analysis, and Kaplan-Meier analysis of best combinations. A five-gene signature was constructed in the discovery cohort (TCGA) and validated in four validation cohorts (GEO). Risk score of the signature stratified patients into high-risk (HRisk) and low-risk (LRisk) subgroups. Patients in the HRisk group were associated with worse OS than those in the LRisk group in both the TCGA cohort (p<0.0001, HR=2.718, 95%CI:1.872-3.947) and the four GEO cohorts (p<0.05). Multivariate Cox regression analyses confirmed that the risk score served as an independent prognostic factor in both cohorts. The signature's predictive capacity was further demonstrated by the nomogram analysis. Pathway enrichment analysis revealed that immunosuppressive and malignant progression-related pathways were enriched in the HRisk group, including TGF-ß, WNT and ECM pathways. The LRisk group was characterized by immune-active signaling pathways (interferon-gamma, T cell activation, etc.) and higher proportions of anti-tumor immune cells (NK, M1, etc.) while HRisk patients were associated with higher stromal scores and less TCR richness. In conclusion, the signature reveals a close relationship between the anoikis and prognosis and may provide a potential therapeutic target for OV patients.

Overexpression of CpADC from Chinese Cherry (Cerasus pseudocerasus Lindl. 'Manaohong') Promotes the Ability of Response to Drought in Arabidopsis thaliana.

Polyamines (PA) play an important role in the growth, development and stress resistance of plants, and arginine decarboxylase (ADC) is one of the key enzymes in the biosynthetic pathway of polyamines. Previously, the transcriptional regulation of the 'Manaohong' cherry under the shelter covering was carried out, and the PA synthase-related genes, particularly the ADC gene, were differentially expressed as exposure to drought stress. However, the mechanisms of how ADC is involved in the response of cherry to abiotic stress (especially drought stress) are still unknown. In the present work, the full-length coding sequence of this gene was isolated and named CpADC. Bioinformatics analysis indicated that the coding sequence of CpADC was 2529 bp in length. Cluster analysis showed that CpADC had the highest homologies with those of sweet cherry (Prunus avium, XP_021806331) and peach (Prunus persica, XP_007200307). Subcellular localization detected that the CpADC was localized in the plant nucleus. The qPCR quantification showed that CpADC was differentially expressed in roots, stems, leaves, flower buds, flowers, and fruits at different periods. Drought stress treatments were applied to both wild-type (WT) and transgenic Arabidopsis lines, and relevant physiological indicators were measured, and the results showed that the putrescine content of transgenic Arabidopsis was higher than that of WT under high-temperature treatment. The results showed that the MDA content of WT was consistently higher than that of transgenic plants and that the degree of stress in WT was more severe than in transgenic Arabidopsis, indicating that transgenic CpADC was able to enhance the stress resistance of the plants. Both the transgenic and WT plants had significantly higher levels of proline in their leaves after the stress treatment than before, but the WT plant had lower levels of proline than that of transgenic Arabidopsis in both cases. This shows that the accumulation of proline in the transgenic plants was higher than that in the wild type under drought and high and low-temperature stress, suggesting that the transgenic plants are more stress tolerant than the WT. Taken together, our results reveal that, under drought stress, the increase in both expressions of CpADC gene and Put (putrescine) accumulation regulates the activity of ADC, the content of MDA and Pro to enhance the drought resistance of Arabidopsis thaliana.

Identification of Genes and Metabolic Pathways Involved in Resin Yield in Masson Pine by Integrative Analysis of Transcriptome, Proteome and Biochemical Characteristics.

Masson pine (Pinus massoniana L.) is one of the most important resin-producing tree species in southern China. However, the molecular regulatory mechanisms of resin yield are still unclear in masson pine. In this study, an integrated analysis of transcriptome, proteome, and biochemical characteristics from needles of masson pine with the high and common resin yield was investigated. The results showed that chlorophyll a (Chl a), chlorophyll b (Chl b), total chlorophyll (Chl C), carotenoids (Car), glucose (Glu), gibberellin A9 (GA9), gibberellin A15 (GA15), and gibberellin A53 (GA53) were significantly increased, whereas fructose (Fru), jasmonic acid (JA), jasmonoyl-L-isoleucine (JA-ILE), gibberellin A1 (GA1), gibberellin A3 (GA3), gibberellin A19 (GA19), and gibberellin A24 (GA24) were significantly decreased in the high resin yield in comparison with those in the common one. The integrated analysis of transcriptome and proteome showed that chlorophyll synthase (chlG), hexokinase (HXK), sucrose synthase (SUS), phosphoglycerate kinase (PGK), dihydrolipoamide dehydrogenase (PDH), dihydrolipoamide succinyltransferase (DLST), 12-oxophytodienoic acid reductase (OPR), and jasmonate O-methyltransferases (JMT) were consistent at the transcriptomic, proteomic, and biochemical levels. The pathways of carbohydrate metabolism, terpenoid biosynthesis, photosynthesis, and hormone biosynthesis may play crucial roles in the regulation of resin yield, and some key genes involved in these pathways may be candidates that influence the resin yield. These results provide insights into the molecular regulatory mechanisms of resin yield and also provide candidate genes that can be applied for the molecular-assisted selection and breeding of high resin-yielding masson pine.

Impact of Tumor-Stroma Ratio on the Prognosis of Colorectal Cancer: A Systematic Review.

BACKGROUND: It is critical to develop a reliable and cost-effective prognostic tool for colorectal cancer (CRC) stratification and treatment optimization. Tumor-stroma ratio (TSR) may be a promising indicator of poor prognosis in CRC patients. As a result, we conducted a systematic review on the predictive value of TSR in CRC. METHODS: This study was carried out according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 guideline. An electronic search was completed using commonly used databases PubMed, CENTRAL, Cochrane Central Register of Controlled Trials, and Google scholar till the last search up to May 30, 2021. STATA version 13 was used to analyze the data. RESULTS: A total of 13 studies [(12 for disease-free survival (DFS) and nine studies for overall survival (OS)] involving 4,857 patients met the inclusion criteria for the systematic review in the present study. In individuals with stage II CRC, stage III CRC, or mixed stage CRC, we observed a significantly higher pooled hazard ratio (HR) in those with a low TSR/greater stromal content (HR, 1.54; 95% CI: 1.20 to 1.88), (HR, 1.90; 95% CI: 1.35 to 2.45), and (HR, 1.70; 95% CI: 1.45 to 1.95), respectively, for predicting DFS. We found that a low TSR ratio had a statistically significant predictive relevance for stage II (HR, 1.43; 95% CI: 1.09 to 1.77) and mixed stages of CRC (HR, 1.65; 95% CI: 1.31 to 2.0) for outcome OS. CONCLUSION: In patients with CRC, low TSR was found to be a prognostic factor for a worse prognosis (DFS and OS).

Prediction of twin pregnancy preeclampsia based on clinical risk factors, early pregnancy serum markers, and uterine artery pulsatility index.

OBJECTIVES: To investigate whether a combination of clinical risk factors, early pregnancy serum markers, and uterine artery pulsatility index (UTPI) can be used to predict twin preeclampsia (PE). METHODS: This case control study included women with twin pregnancies who had undergone obstetrics treatments and gave birth at the Huzhou Maternity and Child Health Care Hospital from October 2018 to November 2020. Patients with PE comprised study group, and patients without PE comprised control group based on selection criteria and a 1:1 ratio. Statistical analysis was performed using clinical risk factors, early pregnancy serum markers, and UTPIs, and the area under the receiver operating curve (AUC. Sensitivity, and the specificity of different combinations of these variables were calculated to predict PE in women with twin pregnancy. RESULTS: Logistic regression analysis revealed four independent predictors for the onset of PE during twin pregnancies: first delivery (OR, 7.51; P=0.045), conception method (OR, 7.11; P=0.036), ß-HCG level (per SD OR, 2.73; P=0.026), and UTPI (OR, 0.17; P=0.043). First-delivery and IVF pregnancy methods both lead to a 7-fold increase in the PE risk during twin pregnancies. Every one sigma (standard deviation) increase in the ß-HCG level led to a 2.73-fold increase in the PE risk. Every UTPI increment by 1.0 reduces the risk of PE by 83%. The prediction efficiencies were based on an AUC of 0.837, a sensitivity of 69%, and a specificity of 92% for the clinical risk factors; an AUC of 0.800, a sensitivity of 81%, and specificity of 78% for the ß-HCG level, and an AUC of 0.814, a sensitivity of 88%, and a specificity of 65% for the UTPI. AUC was 0.928, sensitivity 85%, and a specificity 88% after applying the three types of indicators together for prediction. CONCLUSIONS: By combining early pregnancy serum markers (ß-HCG), and UTPI, the predictive value for PE during twin pregnancy is improved together with its sensitivity and specificity.

Hydroxyapatite-Coated Titanium by Micro-Arc Oxidation and Steam-Hydrothermal Treatment Promotes Osseointegration.

Titanium (Ti)-based alloys are widely used in tissue regeneration with advantages of improved biocompatibility, high mechanical strength, corrosion resistance, and cell attachment. To obtain bioactive bone-implant interfaces with enhanced osteogenic capacity, various methods have been developed to modify the surface physicochemical properties of bio-inert Ti and Ti alloys. Nano-structured hydroxyapatite (HA) formed by micro-arc oxidation (MAO) is a synthetic material, which could facilitate osteoconductivity, osteoinductivity, and angiogenesis on the Ti surface. In this paper, we applied MAO and steam-hydrothermal treatment (SHT) to produce HA-coated Ti, hereafter called Ti-M-H. The surface morphology of Ti-M-H1 was observed by scanning electron microscopy (SEM), and the element composition and the roughness of Ti-M-H1 were analyzed by energy-dispersive X-ray analysis, an X-ray diffractometer (XRD), and Bruker stylus profiler, demonstrating the deposition of nano-HA particles on Ti surfaces that were composed of Ca, P, Ti, and O. Then, the role of Ti-M-H in osteogenesis and angiogenesis in vitro was evaluated. The data illustrated that Ti-M-H1 showed a good compatibility with osteoblasts (OBs), which promoted adhesion, spreading, and proliferation. Additionally, the secretion of ALP, Col-1, and extracellular matrix mineralization was increased by OBs treated with Ti-M-H1. Ti-M-H1 could stimulate endothelial cells to secrete vascular endothelial growth factor and promote the formation of capillary-like networks. Next, it was revealed that Ti-M-H1 also suppressed inflammation by activating macrophages, while releasing multiple active factors to mediate osteogenesis and angiogenesis. Finally, in vivo results uncovered that Ti-M-H1 facilitated a higher bone-to-implant interface and was more attractive for the dendrites, which promoted osseointegration. In summary, MAO and SHT-treated Ti-M-H1 not only promotes in vitro osteogenesis and angiogenesis but also induces M2 macrophages to regulate the immune environment, which enhances the crosstalk between osteogenesis and angiogenesis and ultimately accelerates the process of osseointegration in vivo.

Composite Coating of Graphene Oxide/TiO2 Nanotubes/HHC-36 Antibacterial Peptide Construction and an Exploration of Its Bacteriostat and Osteogenesis Effects.

Graphene oxide (GO), a kind of polymer, is often selected as a controlled released agent, whereas titanium dioxide (TiO2) nanotubes are commonly used as a drug-coated carrier. This study was conducted to develop methods for manufacturing the GO/TiO2/HHC-36 composite coating and exploring its bacteriostat and osteogenesis properties. The GO/TiO2 nanotubes were prepared by electrochemical methods and HHC-36 was then adsorbed to GO/TiO2to obtain GO/TiO2/HHC-36. Sustained release of HHC-36 was analyzed and the antibacterial effect was examined by the inhibition zone test. The biocompatibility and osteogenesis in vitro of GO/TiO2/HHC-36 were explored. Finally, the osteogenesic property of the composite coating was investigated in a rat femoral defect model in vivo. GO/TiO2/HHC-36 was successfully prepared and had good controlled released performance in vitro. The inhibit zone size of S. aureus was 2.1 mm and that of E. coli was 3.0 mm. GO/TiO2/HHC-36 showed good biocompatibility with mesenchymal stem cells (MSCs) and promoted their adhesion, migration, and differentiation. In addition, the secretion of alkaline phosphatase, collagen, mineralized matrix and osteoblast-related nutrient factors of MSCs was increased after treatment with GO/TiO2/HHC-36. Furthermore, GO/TiO2/HHC-36 also stimulated endotheliocytes to secrete VEGF, leading to angiogenesis. Finally, implantation of GO/TiO2/HHC-36 in the rat femur defect model resulted in MSC migration and increased expression of osteoblast related proteins. The composite coating with controlled released of HHC-36 showed distinct antibacterial properties and promoted osteogenesis in vitro and in vivo.

The association between thromboxane A2 receptor gene polymorphisms and the risk of cerebral infarction.

To explore the association between thromboxane A2 receptor (TXA2R) gene polymorphisms and the risk of cerebral infarction. We screened the relevant publications through the search engines in PubMed, Google Scholar, Embase, Web of Science, and China National Knowledge Infrastructure (the latest search update was performed on July 1, 2020). Gene-disease associations were measured using the estimation of OR (95 % CI) based on five genetic inheritance models. Totally three studies were included in this meta-analysis. TXA2R rs768963 polymorphism in homozygote comparison (OR = 1.86, 95 % CI: 1.35-2.56), heterozygote comparison (OR = 1.81, 95 % CI: 1.37-2.39), and dominant model (OR = 1.82, 95 % CI: 1.39-2.37) emerged as risk factors for cerebral infarction. Besides, an increased cerebral infarction risk was observed in the heterozygote comparison (OR = 1.39, 95 % CI: 1.03-1.88) for TXA2R rs2271875 polymorphism. None of the five models showed any association between TXA2R rs4523 polymorphism and cerebral infarction risk. In conclusion, this is the first meta-analysis verifying that TXA2R rs768963 polymorphism and TXA2R rs2271875 polymorphism may be associated with the risk of cerebral infarction.

The prognostic and diagnostic values of MicroRNA-10b in gastric cancer: A comprehensive study based on meta-analysis and TCGA database.

We conducted a study to evaluate the prognostic and diagnostic values of microRNA-10b (miR-10b) in gastric cancer (GC) based on meta-analysis and TCGA database. Relevant studies were searched in English and Chinese database and meta-analysis was conducted on Stata 12.0. The expression value of miR-10b and clinical parameters of GC patients were downloaded from TCGA database, and relevant analyses were conducted on SPSS. High expression of miR-10b was linked with unfavorable overall survival (OS) in GC (HR = 1.572, 95% CI: 1.240-1.992, P < .001). However, the meta-analysis was significant for patients in early stage, but not for patients in advanced stage. The expression of miR-10b-3p was significantly lower in cancer tissue compared with adjacent tissue (P < .001). Meanwhile, the area under the ROC curve (AUC) value was 0.652 (0.562-0.742), P = .001. Disease-free survival analysis showed increasing miR-10b-5p was correlated with worse survival outcome (HR = 2.366, 95% CI: 1.414-3.959, P = .001). In conclusion, miR-10b acts as a tumor suppressor with prognostic and diagnostic values for GC.

Effects of vegetation type on the microbial characteristics of the fissure soil-plant systems in karst rocky desertification regions of SW China.

In karst regions, shallow karst fissure (SKF) soil has proven to be an important plant habitat and soil resource. However, how plants affect the microbial abundance and community composition of SKF soil remains poorly studied. We explored the soil microbial community structure differences in fractured soil-plant systems by determining phospholipid fatty acid (PLFA) profiles under three vegetation types (herbs, shrubs and trees) in SKF and used a bare SKF without vegetation as the control in a karst rocky desertification area. The total microbial biomass and microbial community composition differed between surface soil and SKF soil. The total microbial biomass in surface soil was higher than that in SKF soil. In addition, in contrast to surface soil, the microbial communities in SKF soil were more vulnerable to the effects of environmental variables. Furthermore, plants had a significant positive effect on the accumulation of microbial biomass in surface and SKF soil: shrubs had the strongest effect, followed by trees. Vegetation types significantly affected the ratios of saturated PLFAs to monounsaturated PLFAs (SAT/MONO ratio) and cyclopropyl PLFAs to precursors (cy/pre ratio). In contrast to the SKF without vegetation, the SAT/MONO ratio and cy/pre ratio under grasslands, shrublands and trees were low. Herbs and shrubs had the greatest capacity to enhance the ability of soil to respond to environmental stress compared to trees. Our results suggest that, as an important plant habitat in karst regions, the condition of SKF soil should be urgently improved. The stereoscopic collocation of shrub-grass vegetation may be the preferred measure for vegetation restoration. Deep-rooted shrubs and grasses are best at improving soil and plant growth. Our study can be useful for developing strategies for vegetation rehabilitation in karst regions.

Prognostic and clinicopathologic significance of MicroRNA-125a-5p in cancers: A meta-analysis.

The aim of the study was to estimate the prognostic and clinicopathologic significance of miR-125a-5p in human cancers. Eligible studies were obtained from PubMed, Embase, and the Cochrane Library. Combined hazard ratios (HRs) and odds ratios (ORs) were used to evaluate the prognostic and clinicopathologic value of miR-125a-5p. In pan-cancer, high miR-125a-5p expression was associated with better overall survival (OS) (HR = 0.459, 95% confidence interval [CI]: 0.369-0.57, P < .001), and disease-free survival (HR = 0.343, 95% CI: 0.237-0.496, P < .001). Furthermore, favorable OS was also found in lung cancer (HR = 0.343, 95% CI: 0.228-0.517, P < .001) and gastric cancer (HR = 0.341, 95% CI: 0.160-0.725, P = .005) patients with high miR-125a-5p expression. Besides, high miR-125a-5p expression was correlated with early stage (OR = 0.413, 95% CI: 0.228-0.749, P = .004) and negative lymph node metastasis (OR = 0.262, 95% CI: 0.073-0.941, P = .04) in gastric cancer, and was linked with better tumor differentiation in pan-cancer (OR = 1.623, 95% CI: 1.064-2.476, P = .025) and lung cancer (OR = 2.371, 95% CI: 1.358-4.141, P = .002). In conclusion, miR-125a-5p is a tumor suppressor with prognostic and clinicopathologic values for human cancer, and miR-125a-5p overexpression predicted favorable prognosis, early stage, negative lymph node metastasis, and better tumor differentiation. More research should be conducted to test these results.

Global transcriptome and gene co-expression network analyses on the development of distyly in Primula oreodoxa.

Distyly is a genetically controlled flower polymorphism that has intrigued both botanists and evolutionary biologists ever since Darwin's time. Despite extensive reports on the pollination and evolution of distylous systems, the genetic basis and mechanism of molecular regulation remain unclear. In the present study, comparative transcriptome profiling was conducted in primrose (Primula oreodoxa), the prime research model for heterostyly. Thirty-six transcriptomes were sequenced for styles at different stages and corolla tube in the three morphs of P. oreodoxa. Large numbers of differentially expressed genes (DEGs) were detected in the transcriptomes of styles across different morphs. Several transcription factors (TFs) and phytohormone metabolism-related genes were highlighted in S-morphs. A growing number of genes showed differential expression patterns along with the development of styles, suggesting that the genetic control of distyly may be more complicated than ever expected. Analysis of co-expression networks and module-trait relationships identified modules significantly associated with style development. CYP734A50, a key S-locus gene whose products degrade brassinosteroids, was co-expressed with many genes in the module and showed significant negative association with style length. In addition, crucial TFs involved in phytohormone signaling pathways were found to be connected with CYP734A50 in the co-expression module. Our global transcriptomic analysis has identified DEGs that are potentially involved in regulation of style length in P. oreodoxa, and may shed light on the evolution and broad biological processes of heterostyly.

Reconstruction using massive allografts after resection of extremity osteosarcomas the study design: A retrospective cohort study.

PURPOSE: Allografts have been shown useful in the reconstruction of bone defects after tumor resection. This study aimed to investigate the feasibility of using massive allografts to reconstruct bone defects after resection of extremity osteosarcomas. METHODS: The clinical data of 15 patients treated with massive allograft reconstruction after resection of extremity osteosarcomas from January 2005 to January 2008 were retrospectively reviewed. Neoadjuvant and postoperative chemotherapy was used in all patients. The postoperative functions of the salvaged limbs were evaluated using the scoring system proposed by the Musculoskeletal Tumor Society (MSTS). RESULTS: All patients were followed up for a mean of 61 months (range, 14-99 months). No nonunion occurred during follow-up. The mean time to union was 9 months (range, 3-21 months). No immune rejection, allograft infection, allograft fracture, and limb length disparity occurred. However, 2 patients had broken implants. The mean MSTS score at the last follow-up was 26 points. Four patients died and 2 patients had tumor recurrence. The 5-year disease free survival rate was 73.3%. CONCLUSION: Massive allograft reconstruction is safe and effective for bone defects caused by resection of extremity osteosarcomas.