New Perspectives and Prospects of microRNA Delivery in Diabetic Wound Healing.

The remarkable potential of microRNAs (miRNAs) as a class of biotherapeutic agents in the treatment of diverse pathological conditions has garnered significant interest in recent years. To heal both acute and chronic wounds, miRNAs work by post-transcriptionally controlling various proteins and the pathways that are linked to them. Diabetes mellitus predisposes to several macro- and microvascular defects of end organs such as atherosclerosis, peripheral artery disease, retinopathy, nephropathy, neuropathy, and impaired wound healing. Here, miRNAs emerge as a beacon of hope, with the capacity to heal diabetic wounds by precisely modulating the expression of genes involved in the healing process. Despite the therapeutic promise, the journey to realizing the full potential of miRNAs is fraught with challenges. Their intrinsic instability and the inefficient delivery into target cells pose significant barriers to their clinical application. Consequently, a major focus of current research is the discovery of novel miRNAs and the development of innovative delivery systems that can effectively transport these nucleic acids into the cells where they are needed most. This review delves into the intricate roles that miRNAs play at various stages of diabetic wound healing, providing a comprehensive overview of the latest research findings. The review also addresses the obstacles and opportunities that come with translating miRNA-based strategies into clinical practice, offering a critical assessment of the field's advancements and the hurdles that remain to be overcome. SIGNIFICANCE STATEMENT: The potential of microRNA delivery using new biological or nonbiological carriers may create a revolutionary treatment method for chronic wounds of diabetes.

Silibinin attenuates TGF-ß2-induced fibrogenic changes in human trabecular meshwork cells by targeting JAK2/STAT3 and PI3K/AKT signaling pathways.

Transforming growth factor-ß2 (TGF-ß2) induced fibrogenic changes in human trabecular meshwork (HTM) cells have been implicated in trabecular meshwork (TM) damage and intraocular pressure (IOP) elevation in primary open-angle glaucoma (POAG) patients. Silibinin (SIL) exhibited anti-fibrotic properties in various organs and tissues. This study aimed to assess the effects of SIL on the TGF-ß2-treated HTM cells and to elucidate the underlying mechanisms. Our study found that SIL effectively inhibited HTM cell proliferation, attenuated TGF-ß2-induced cell migration, and mitigated TGF-ß2-induced reorganization of both actin and vimentin filaments. Moreover, SIL suppressed the expressions of fibronectin (FN), collagen type I alpha 1 chain (COL1A1), and alpha-smooth muscle actin (α-SMA) in the TGF-ß2-treated HTM cells. RNA sequencing indicated that SIL interfered with the phosphoinositide 3-kinase (PI3K)/protein kinase B (PKB, also known as AKT) signaling pathway, extracellular matrix (ECM)-receptor interaction, and focal adhesion in the TGF-ß2-treated HTM cells. Western blotting demonstrated SIL inhibited the activation of Janus kinase 2 (JAK2)/signal transducers and activators of transcription 3 (STAT3) and the downstream PI3K/AKT signaling pathways induced by TGF-ß2, potentially contributing to its inhibitory effects on ECM protein production in the TGF-ß2-treated HTM cells. Our study demonstrated the ability of SIL to inhibit TGF-ß2-induced fibrogenic changes in HTM cells. SIL could be a potential IOP-lowering agent by reducing the fibrotic changes in the TM tissue of POAG patients, which warrants further investigation through additional animal and clinical studies.

Retinoic acid released from self-assembling peptide activates cardiomyocyte proliferation and enhances repair of infarcted myocardium.

The limited cardiomyocyte proliferation is insufficient for repair of the myocardium. Therefore, activating cardiomyocyte proliferation might be a reasonable option for myocardial regeneration. Here, we investigated effect of retinoic acid (RA) on inducing adult cardiomyocyte proliferation and assessed efficacy of self-assembling peptide (SAP)-released RA in activating regeneration of the infarcted myocardium. Effect of RA on inducing cardiomyocyte proliferation was examined with the isolated cardiomyocytes. Expression of the cell cycle-associated genes and paracrine factors in the infarcted myocardium was examined at one week after treatment with SAP-carried RA. Cardiomyocyte proliferation, myocardial regeneration and improvement of cardiac function were assessed at four weeks after treatment. In the adult rat myocardium, expression of RA synthetase gene Raldh2 and RA concentration were decreased significantly. After treatment with RA, the proliferated cardiomyocytes were increased. The formulated SAP could sustainedly release RA. After treatment with SAP-carried RA, expression of the pro-proliferative genes in cell cycle and paracrine factors in the infarcted myocardium were up-regulated. Myocardial regeneration was enhanced, and cardiac function was improved significantly. These results demonstrate that RA can induce adult cardiomyocytes to proliferate effectively. The sustained release of RA with SAP is a promise strategy to enhance repair of the infarcted myocardium.

First report of Diaporthe eres causing leaf spot on Rhododendron latoucheae Franch. in China.

Rhododendron latoucheae Franch. is an evergreen shrub with charming fragrance and large and abundant flowers that make it highly attractive as an ornamental species. The species is distribution in southwest China covers several different habitats and environments (Zhang, et al. 2022). From May to July in 2023, symptoms of leaf spot were observed on R. latoucheae over a wide portion of the Baili Azalea Forest Area (27°10' to 27°20'N, 105°04' to 106°04'E), Guizhou Province, China. About 500 plants were surveyed, and the incidence of leaf spot on R. latoucheae leaves was 12%, significantly reducing their ornamental and economic value. The affected leaves had irregular, grey white lesions with a clear blackish brown boundary and faint black conidiomata in a brown center. To isolate the pathogen, 15 symptomatic leaves were collected from 10 plants. A few black dots were picked from the lesions with a sterilized needle, plated on water agar, and incubated at 25°C for 24 h to observe spore germination (Choi et al. 1999). Then the germinated spores were transferred onto PDA for further purification and morphological observation. Three single-spore isolates (GULJ1-L7, GULJ1-L8, and GULJ1-L9) identical in morphology were obtained. The isolate GULJ 1-L7 was used for further study. Colonies on PDA irregular grew white felty aerial mycelium, becoming white felted aerial mycelium in the centre and grey-brown mycelium at the marginal area on the upper surface, while the lower surface presents alternating white, tan and taupe. Colony with conidiomata irregularly distributed over agar surface. In the representative isolate, darkly pigmented pycnidia (flask-shaped) were produced over the colony surface on PDA after about 30 days, and oozed milky or yellowish mucilaginous drops. The fungus produced two types of conidia, α and ß. Regular α conidia were 5.15-10.29 × 1.54-3.33 µm (n = 50), hyaline, elongated, biguttulate and non-septate. Beta conidia were 20.34-31.91 × 1.01-1.90 µm (n = 50), aseptate, hyaline, smooth, spindle shaped, slightly curved to bent. The morphological features were consistent with the description of Diaporthe eres (Pereira, et al. 2022). The pathogen was confirmed to be D. eres by amplification and sequencing of the internal transcribed spacer region (ITS), the partial ß-tubulin (TUB), the partial translation elongation factor 1-alpha (TEF) genes and the calmodulin (CAL) using primers ITS1/ITS4, Bt-2a/Bt-2b, EF1-728F/EF1-986R, and CAL-228F/CAL-737R, respectively (Tao et al. 2020). Sequences from PCR amplification were deposited in GenBank with accession numbers OR740563 (ITS), OR754301 (TUB), OR754298 (TEF), and OR754295 (CAL) respectively. BLAST searches of the sequences revealed 99.07% (533/538 nt), 100% (490/490 nt), 99.69% (317/318 nt) and 98.95% (376/380 nt) homology with those of D. eres AR5193T from GenBank (KJ210529.1, KJ420799.1, KJ210550.1 and KJ434999.1), respectively. Phylogenetic analysis (MEGA 7.0) using the maximum-likelihood method placed the isolate GULJ1-L7 in a well-supported cluster with D. eres CBS 138694T. The pathogen was thus identified as D. eres based on the morphological characterization and molecular analyses (Feng, et al. 2013; Tao, et al. 2020). The pathogenicity of GULJ1-L7 was tested through a pot assay. Due to the difficulty of artificial planting wild R. latoucheae, we conducted a pot essay to detect the pathogenicity of GULJ1-L7 using a closely related Rhododendron delavayi Franch. Ten healthy R. delavayi plants were scratched with a sterilized needle (0.45 mm in diameter) on three leaves per plant. Plants were inoculated by spraying α and ß spore mixture suspension (106 spores ml-1) of GULJ1-L7 onto leaves until runoff, and the control leaves were sprayed with sterile water. The plants were maintained at 25°C and 75% relative humidity in a growth chamber. The pathogenicity test was repeated three times. After 14 days, the treated leaves developed brown lesions similar to those in the field, whereas the control had no symptoms. The same fungus was reisolated from the infected leaves and identified based on a morphological characterization and molecular analyses. These results fulfilled Koch's postulates. To our knowledge, this is the first report of leaf spot on R. latoucheae caused by D. eres in China. The fungal pathogen identification will provide valuable information for prevention and management of leaf spot disease associated with R. latoucheae.

Niclosamide inhibits TGF-ß1-induced fibrosis of human Tenon's fibroblasts by regulating the MAPK-ERK1/2 pathway.

Preventing postoperative bleb scar formation is an effective way of improving glaucoma filtration surgery (GFS) outcome. Use of more effective antifibrotic drugs with fewer adverse effects may be a good way to address the problem. In the present study, we use a primary cell model, consisting of Tenon's fibroblasts obtained from patients with glaucoma, which were stimulated with TGF-ß1 to induce the fibrotic phenotype. We explored the effects of niclosamide on TGF-ß1-induced fibrosis in these cells and examined its underlying mechanism of action. A transcriptome sequencing assay was used to explore possible signaling pathways involved. Niclosamide inhibited cell proliferation and migration, and decreased the levels of alpha-smooth muscle actin, type I and type III collagen in human Tenon's fibroblasts induced by TGF-ß1. Niclosamide also induced apoptosis and counteracted TGF-ß1-induced cytoskeletal changes and extracellular matrix accumulation. Moreover, niclosamide decreased TGF-ß1-induced phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) protein expression in human Tenon's fibroblasts. The results indicate that niclosamide inhibits TGF-ß1-induced fibrosis in human Tenon's fibroblasts by blocking the MAPK-ERK1/2 signaling pathway. Thus, niclosamide is a potentially promising antifibrotic drug that could improve glaucoma filtration surgery success rate.

Serelaxin Inhibits Lipopolysaccharide-induced Inflammatory Response in Cardiac Fibroblasts by Activating Peroxisome Proliferator-activated Receptor-γ and Suppressing the Nuclear Factor-Kappa B Signaling Pathway.

ABSTRACT: Serelaxin (sRLX) has an inhibitory effect on fibrosis. However, whether the antifibrotic effects of sRLX are achieved by inhibiting the inflammatory response has not been clarified. This study aimed to investigate the role of sRLX in lipopolysaccharide (LPS)-induced inflammation in cardiac fibroblasts and elucidate the underlying mechanisms. Cardiac fibroblasts were isolated from adult rat hearts. The effect of sRLX on the inhibition of the inflammatory response after LPS induction was examined. Cell viability was measured by MMT assay. Cell proliferation was determined using the Cell Counting Kit-8. The levels of inflammatory cytokines IL-1ß, IL-6, TNF-α, and IL-10 were measured using an enzyme-linked immunosorbent assay. The mRNA levels of α-smooth muscle actin (α-SMA), collagen I/III, MMP-2, MMP-9, IL-1ß, IL-6, TNF-α, IL-10, IκBα, p-IκBα, p65 subunit of nuclear factor-kappa B (NF-κB), and peroxisome proliferator-activated receptor-γ (PPAR-γ) were assessed by real-time quantitative PCR. The protein levels of α-SMA, collagen I/III, MMP-2, MMP-9, IκBα, p-IκBα, p65, p-p65, and PPAR-γ were examined by western blotting. sRLX inhibited LPS-induced IL-1ß, IL-6, TNF-α, α-SMA, and collagen I/III, and elevated the expression of IL-10, MMP-2, and MMP-9. Moreover, LPS-induced activation of NF-κB pathway was suppressed by sRLX treatment. Further studies showed that sRLX did not significantly increase the expression of PPAR-γ mRNA and protein, but activated PPAR-γ activity, and the PPAR-γ inhibitor GW9662 reversed the inhibitory effect of sRLX on IL-1ß, IL-6, and TNF-α production. These results suggest that sRLX alleviates cardiac fibrosis by stimulating PPAR-γ through a ligand-independent mechanism that subsequently abolish the expression of NF-κB signaling pathway.

Helicobacter pylori Infection as a Predictor of Treatment Outcomes of Gastric Cancer: A Systematic Review and Meta-Analysis.

BACKGROUND: Helicobacter pylori infection is strongly associated with gastric cancer. However, there is currently no consensus on the association between H. pylori and gastric cancer prognosis. METHODS: A systematic search was conducted on studies in PubMed, EMBASE, and Web of Science up to March 10, 2022. The quality of all included studies was assessed using the Newcastle-Ottawa Scale. The hazard ratio (HR) and its 95% confidence interval (95% CI) were extracted to analyze the association between H. pylori infection and prognosis of gastric cancer. In addition, subgroup analysis and publication bias were performed. RESULTS: A total of 21 studies were involved. The pooled HR was 0.67 (95% CI, 0.56-0.79) for overall survival (OS) in H. pylori-positive patients, with the control (HR = 1) being the H. pylori-negative group. In the subgroup analysis, the pooled HR was 0.38 (95% CI, 0.24-0.59) for OS in H. pylori-positive patients who received surgery combined with chemotherapy. The pooled HR for disease-free survival was 0.74 (95% CI, 0.63-0.8) and 0.41 (95% CI, 0.26-0.65) in patients who received surgery combined with chemotherapy. CONCLUSION: H. pylori-positive gastric cancer patients have a better overall prognosis than H. pylori-negative patients. H. pylori infection has improved the prognosis of patients undergoing surgery or chemotherapy, among which the improvement was most obvious in patients undergoing surgery combined with chemotherapy.

Effects of Whole-Body Vibration Training on Lower Limb Muscle Strength and Physical Performance Among Older Adults: A Systematic Review and Meta-analysis.

OBJECTIVE: To evaluate the effects of whole-body vibration training (WBVT) on lower limb muscle strength and physical performance in older adults. DATA SOURCES: Web of Science, PubMed, Cochrane Library, and MEDLINE databases were searched for papers published in English, from January 1, 2000, to May 30, 2022. STUDY SELECTION: Randomized controlled trials of WBVT in older adults (mean age, 65 years or older) published in English. The Physiotherapy Evidence Database Scale was used to assess the quality of the selected studies. DATA EXTRACTION: Two investigators independently assessed articles according to the evaluation criteria. Differences between investigator assessments were resolved by consulting a third investigator before reassessment. DATA SYNTHESIS: Systematic review of 18 randomized controlled studies found that WBVT produced significant improvements in lower limb muscle strength and physical performance among older adults. We used the Cochrane Collaboration method to assess risk of bias and RevMan version 5.4a to extract means and calculate SDs. WBVT significantly improved knee strength (standard mean difference [SMD]=0.72, 95% confidence interval [CI] [0.38, 1.07], P<.0001, I2=58%) and explosive power (SMD=0.47, 95% CI [0.10, 0.83], P=.01, I2=0%) among older adults. Significant improvements in each physical performance were observed in the sit-to-stand test (SMD=0.57, 95% CI [0.30, 0.84], P<.0001, I2=35%), the subgroup of studies that evaluated balance with timed Up and Go test, SMD was 0.53 (95% CI [0.19, 0.88], P=.002, I2=56%) and the Tinetti total score, SMD was 0.72 (95% CI [0.04, 1.41], P=.04, I2=81%), walking speed (SMD=0.46, 95% CI [0.14, 0.77], P=.005, I2=49%), and walking endurance (SMD=0.43, 95% CI [0.02, 0.85], P=.04, I2=24%). CONCLUSIONS: WBVT may be an effective intervention to improve lower limb muscle strength and physical performance in older adults. Tinetti total score remains controversial and warrants assessment in future high-quality randomized controlled trials.

First report of Pseudopithomyces chartarum causing leaf spot on Tetrapanax papyrifer in Guizhou, China.

Tetrapanax papyrifer (Hook.) K. Koch, widely utilized in clinical practices in traditional Chinese medicine, is a medicinal plant whose dried stem pich exhibits activities such as lactation induction, diuresis, and anti-inflammatory effects. The species is native to the southwest of China, such as Guizhou and Yunnan provinces. It thrives in sunlight and warmth and is planted in fertile valleys in the region (Zhang et al. 2023). In July 2021, a leaf spot-like disease was observed on approximately 15% of T. papyrifer (Big T. papyrifer) in a field in Shibing County (127.2°E, 25.2°N), Guizhou Province, China. The symptomatic leaves displayed irregular, watery dark brown lesions with black conidiomata in gray centers and surrounded by yellow halos. To identify the causal agent leading to the disease, 15 symptomatic leaves from five trees in one field were collected. These leaves underwent surface sterilization, which included 30s in 75% ethanol, 2 min in 3% NaOCl, and three times of washing with sterilized distilled water. Thereafter, small pieces of the symptomatic leaf tissues (0.2 × 0.2 cm) were plated on PDA and incubated at 25°C for seven days (Fang 2007). Three isolates were obtained based on the improved single spore isolation method proposed by Gong et al. (2010), and named as GUTC 321, GUTC 523 and GUTC 873. The fungal colonies on PDA were villiform, creamy-white, whorled, and sparse aerial mycelium on the surface with black, gregarious conidiomata. The conidia were ellipsoid, mid brown to dark brown, mainly with 3-4 transverse septa, usually divided by longitudinal septum, often constricted at the septa, 21.8 (12.6-34.5) × 13.9 (8.8-19.8) µm (n=50). The morphological features were consistent with the descriptions of Pseudopithomyces chartarum (Ariyawansa et al. 2015). All three isolates exhibited identical morphological properties. The potential pathogen was confirmed as P. chartarum by amplification and sequencing of the internal transcribed spacer regions (ITS), large subunit ribosomal (LSU) and translation elongation factor 1 alpha (TEF1) genes with primers ITS4/ITS5, LROR/LR7 and EF-983F/EF-2218R, respectively (Ariyawansa et al. 2015; Jayasiriet al. 2019). BLASTn analyses of the sequences showed 100% identity among the three isolates and a high homology (ITS, 99.8%: 598/599; LSU, 100%: 853/853; and TEF1, 100%: 871/871) with those of P. chartarum sequences in GenBank (MT123059, OK655822, and MK360080, respectively). The sequences of the genes from isolate GUTC321 were deposited in GenBank under accession numbers OP269599 (ITS), OP237015 (LSU), and OR069689 (TEF1). Phylogenetic analyses of the concatenated ITS-LSU-TEF1 sequence (2,685 bp) of GUTC 321 using PhyloSuite 1.2.2 with PartitionFinder model revealed that the isolate clustered closely with P. chartarum isolate CBS 329.86T (Cecilia 1986). The pathogenicity of GUTC 321 was tested thereafter on ten healthy T. papyrifer plants grown in pots in growth chamber. The plants were inoculated by spraying with spore suspension (106 spores mL-1) of GUTC 321 or sterile water (control) onto leaves that had been slightly injured with sterilized SiO2 (0.1-0.25 mm) until runoff. The plants were maintained at 25°C in the growth chamber, and monitored for symptom development. Local lesions began to appear on all GUTC 321-inoculated leaves, but not on those of the control plants, 48 hours after inoculation. Seven days after the inoculation, lesions similar to those observed on field plants occurred on GUTC321-inoculated plants but not on the control plants, the lesions observed only in inoculated leaves. The same fungus was reisolated and identified based on the morphological characterization and molecular analyses (ITS, LSU and TEF1) from the infected leaves thus fulfilling Koch's postulates. To our knowledge, this is the first report of leaf spot on T. papyrifer caused by P. chartarum in China. Considering the significance of T. papyrifer in Chinese medicine, approximate management measures need to be developed and applied to control the disease in the crop.

First Report of Anthracnose on Tetrapanax papyriferus Caused by Colletotrichum fructicola in China.

Tetrapanax papyriferus is an evergreen shrub native to China and traditionally used as a herbal medicine (Li et al., 2021). In September 2021, a serious leaf spot disease with symptoms similar to anthracnose was extensively observed on T. papyriferus in Shibing county (E 127°12'0", N 25°11'60"), Qiandongnan Miao and Dong Autonomous Prefecture, Guizhou province, China. Field surveys were conducted in about 1000 T. papyriferus plants in Shibing in September 2021. The incidence of the leaf spot on leaves was 45% to 60%, significantly reducing the quality of medicinal materials. The symptoms began as small yellow spots, developing a brown center and dark brown to black margin, and eventually the diseased leaves were wiltered and rotted. Symptomatic leaves were collected from 20 trees. Symptomatic tissue from diseased leaves was surface desinfected (0.5 min in 75% ethanol and 1 min in 3% NaOCl, washed three times with sterilized distilled water), small pieces of symptomatic leaf tissue (0.2 × 0.2 cm) were plated on potato dextrose agar (PDA) and incubated at 25°C for about 7 days (Fang. 2007). Three single-spore isolates were obtained (GUTC37, GUTC310 and GUTC311) and deposited in the collection of the Plant Pathology Deparment, College of Agriculture, Guizhou University, China (GUCC) (with the accession numbers, GUCC220241, GUCC220242, GUCC220243 respectively). These isolates were identical in morphology and in the sequences of internal transcribed spacer region [ITS], glyceraldehy-3-phosphate dehydrogenase [GAPDH], chitin synthase [CHS-1], actin [ACT], and calmodulin [CAL] genes (White et al. 1990; Carbone and Kohn 1999; Templeton et al. 1992). Therefore, the representative isolate GUTC37 was used for further analysis. The pathogenicity of GUTC37 was tested through a pot assay. Plants were inoculated by spraying a spore suspension (106 spores·ml-1) of isolated strains onto leaves until runoff, and the control leaves sprayed with sterile water. The inoculated plants were incubated in a growth chamber at 28 ℃ and 95% relative humidity for 10 days. Pathogenicity tests were repeated three times (Fang. 2007). The symptoms developed on the inoculated leaves, while control remained asymptomatic. The lesions were first visible 72 h after inoculation, and typical lesions like those observed on field plants appeared after 10 days. The same fungus was reisolated and identified based on the morphological characterization and molecular analyses from the infected leaves but not from the non-inoculated leaves. Results of pathogenicity experiments of isolated fungi fulfilled Koch's postulates. Fungal colonies on PDA were villiform, creamy-white or greyish, aerial mycelium pale grey, dense, surface partly covered with orange conidial masses. The conidia were abundant, oval-ellipsoid, aseptate, and 13.89 (11.62 to 15.21) × 5.21 (4.39 to 5.65) µm (n=50). Appressorium were greyish green, nearly ovoid to cylindrical, 9.64 (6.62 to 14.61) × 6.33 (5.45-7.72) µm (n=50). The morphological features were consistent with the descriptions of Colletotrichum fructicola Prihast., L. Cai & K.D. Hyde (Prihastuti et al. 2009). The pathogen was identified to be C. fructicola by amplification and sequencing of the five genes. The sequences of the PCR products were deposited in GenBank with accession numbers OP143657 (ITS), OP177868 (GAPDH), OP177865 (CHS-1), OP278677 (ACT) and OP177862 (CAL). BLAST searches of the obtained sequences revealed 100% (509/509 nucleotides), 99.63% (269/270 nucleotides), 99.31% (287/289 nucleotides), 99.29% (280/282 nucleotides), and 99.86% (728/729 nucleotides) homology with those of C. fructicola in GenBank (JX010165, JX010033, JX009866, FJ907426, and JX009676, respectively). Phylogenetic analysis (MEGA 7.0) using the maximum likelihood method placed the isolate GUTC37 in a well-supported cluster with C. fructicola. To our knowledge, this is the first report of anthracnose on T. papyriferus caused by C. fructicola in Guizhou, China. This study provides valuable information for the identification and control of the anthracnose on T. papyriferus.

First Report of Grey Spot on Tobacco caused by Alternaria alstroemeriae in China.

Tobacco (Nicotiana tabacum L.) is an important economic crop belonging to family Solanaceae and is widely cultivated in China (Basit 2021). From April to July in 2022, a foliar disease with symptoms similar to grey spot was extensively observed on tobacco in Guangxi Province (24°52' N, 111°23' E), China. Field surveys were conducted in 18 towns and the disease incidence was 0.89% to 6.95%. Symptomatic leaves displayed irregular, dark brown lesions surrounded by yellow halos and accompanied with black conidiomata in gray centers (Fig 1A-E). Symptomatic leaves were collected from 54 different tobacco plants. After surface sterilization (0.5 min in 75% ethanol and 1 min in 3% NaOCl, washed three times with sterilized distilled water), small pieces of symptomatic leaf tissue (0.2 × 0.2 cm) were plated on PDA and incubated at 25°C for 5 days (Fang 2007). Three single-spore isolates, GUCC BZ6-3, GUCC LJ3-4, and GUCC XH1-13 were obtained, which were identical in morphology and molecular analysis. Therefore, the representative isolate GUCC BZ6-3 was used for further study. The colonies on PDA were villiform, greyish (Fig 1F-G). Conidia were abundant, ovoid, with 2-6 transverse septa and 1-2 longitudinal septa 12.60 (9.43 to 14.76) × 4.30 (3.57 to 5.14) µm (n=50) (Fig 1H-S). The morphological features were consistent with Alternaria alstroemeriae E.G. Simmons & C.F. Hill (Simmons 2007; Nishikawa & Nakashima, 2013). The pathogen was confirmed to be A. alstroemeriae by amplification and sequencing of the ITS, GAPDH, LSU, TEF1, and RBP2 genes using primers ITS1/ITS4, gpd1/gpd2, LSU1Fd/LR5, EF1-728F/EF1-986R, and RPB2-5F2/fRPB2-7cR, respectively (Woudenberg 2013). The sequences of the PCR products were deposited in GenBank with accession numbers ON693856 (RBP2), ON714497 (ITS), ON694345 (GAPDH), ON931420 (TEF1) and ON714499 (LSU). BLAST searches of the obtained sequences revealed 99% (565/567 nucleotides), 99% (577/579 nucleotides), 99% (908/911 nucleotides), 99% (238/239 nucleotides), and 99% (751/753 nucleotides) homology with those of A. alstroemeriae in GenBank (MH863036, KP124154, MH874589, KP125072, and KP124765, respectively). Phylogenetic analyses of the sequence data consisted of Bayesian and Maximum likelihood analyses of the combined aligned dataset (MEGA 7.0 and PhyloSuite 1.2.2). The GUCC BZ6-3 in a well-supported cluster with A. alstroemeriae (Fig 2). The pathogen was thus identified as A. alstroemeriae based on morphological characterization and molecular analyses. The pathogenicity of GUCC BZ6-3 was tested through pot assay and carried out three times (Fang 2007). Ten healthy 30-day-old tobacco plants were inoculated by spraying a spore suspension (106 spores·ml-1) of strain GUCC BZ6-3 onto leaves until runoff, and the control leaves were sprayed with sterile water. The plants were maintained at 28°C with high relative humidity (95%) in a growth chamber. The symptoms developed on all inoculated leaves but not on the control. The lesions were first visible 48 h after inoculation, and typical lesions similar to those observed on field plants appeared after 7 days. The same fungus was reisolated and identified based on the morphological characterization and molecular analyses from the infected leaves but not from the noninoculated leaves. Results of pathogenicity experiments fulfilled Koch's postulates. To our knowledge, this is the first report of grey spot disease on tobacco caused by A. alstroemeriae in China. Our findings would be of great importance for the diagnosis and control of the emerging grey spot on tobacco.

Partial substitution of manure reduces nitrous oxide emission with maintained yield in a winter wheat crop.

Conventional fertilization of agricultural soils results in increased N2O emissions. As an alternative, the partial substitution of organic fertilizer may help to regulate N2O emissions. However, studies assessing the effects of partial substitution of organic fertilizer on both N2O emissions and yield stability are currently limited. We conducted a field experiment from 2017 to 2021 with six fertilizer regimes to examine the effects of partial substitution of manure on N2O emissions and yield stability. The tested fertilizer regimes, were CK (no fertilizer), CF (chemical fertilizer alone, N 300 kg ha-1, P2O5 150 kg ha-1, K2O 90 kg ha-1), CF + M (chemical fertilizer + organic manure), CFR (chemical fertilizer reduction, N 225 kg ha-1, P2O5 135 kg ha-1, K2O 75 kg ha-1), CFR + M (chemical fertilizer reduction + organic manure), and organic manure alone (M). Our results indicate that soil N2O emissions are primarily regulated by soil mineral N content in arid and semi-arid regions. Compared with CF, N2O emissions in the CF + M, CFR, CFR + M, and M treatments decreased by 16.8%, 23.9%, 42.0%, and 39.4%, respectively. The highest winter wheat yields were observed in CF, followed by CF + M, CFR, and CFR + M. However, the CFR + M treatment exhibited lower N2O emissions while maintaining high yield, compared with CF. Four consecutive years of yield data from 2017 to 2021 illustrated that a single application of organic fertilizer resulted in poor yield stability and that partial substitution of organic fertilizer resulted in the greatest yield stability. Overall, partial substitution of manure reduced N2O emissions while maintaining yield stability compared with the synthetic fertilizer treatment during the wheat growing season. Therefore, partial substitution of manure can be recommended as an optimal N fertilization regime for alleviating N2O emissions and contributing to food security in arid and semi-arid regions.

Comparison of the Effectiveness and Safety of Different Methods of Colorectal Endoscopic Submucosal Dissection: A Systematic Review and Network Meta-Analysis.

BACKGROUND: Traditional endoscopic submucosal dissection (ESD) has developed different methods, such as pocket method (P-ESD), traction-assisted method (T-ESD), and hybrid method (H-ESD). In this meta-analysis, the benefits and drawbacks of different ESD methods were discussed and ranked. STUDY DESIGN: Studies comparing different methods of colorectal ESD were searched by using PubMed, EMBASE, and Cochrane Library databases. The study was conducted for five endpoints: en bloc resection rate, R0 resection rate, operation time, dissection speed, and adverse events rate. Pairwise and network meta-analyses were performed through Rev Man 5.4 and Stata 16.0. The quality of all included studies was assessed using the Cochrane Risk of Bias Tool and the Newcastle-Ottawa Scale. RESULTS: Twenty-six studies met the inclusion criteria, including 7 RCTs and 19 non-RCTs, with a total of 3,002 patients. The pooled analysis showed that the en bloc resection rate of H-ESD was significantly lower than that of C-ESD, P-ESD, and T-ESD (RR = 0.28, 95% CI [0.12, 0.65]; RR = 0.11, 95% CI [0.03, 0.44]; RR = 8.28, 95% CI [2.50, 27.42]). Compared with C-ESD, the operation time of H-ESD and T-ESD was significantly shorter (MD = -21.83, 95% CI [-34.76, -8.90]; MD = -23.8, 95% CI [-32.55, -15.06]). Meanwhile, the operation time of T-ESD was also significantly shorter than that of P-ESD (MD = -18.74, 95% CI [-31.93, -5.54]). The dissection speed of T-ESD was significantly faster than that of C-ESD (MD = 6.26, 95% CI [2.29, 10.23]). CONCLUSION: P-ESD and T-ESD are probably the two best methods of colorectal ESD at present. The advantages of P-ESD are high en bloc resection rate and low incidence of adverse events. The advantages of T-ESD are rapid dissection and short operation time.

Endoscopic Therapy of Small Bowel Polyps by Single-Balloon Enteroscopy in Patients with Peutz-Jeghers Syndrome.

Background: Little is known about the efficacy and safety of single-balloon enteroscopy (SBE) in patients with Peutz-Jeghers syndrome (PJS). The aim of this study was to assess the efficacy and safety of SBE for the treatment of small bowel polyps in patients with PJS. Methods: We conducted a single-center observational study, which included all patients diagnosed with PJS who underwent SBE for polypectomy between January 2018 and March 2021. Complete treatment was defined as the absence of polyps ≥10 mm after SBE resection. The clinical records were retrospectively reviewed. Results: 102 patients (including 40 men and 62 women) with a mean age of 28.7 years (range 13-55 y) were enrolled in our study. The intubation depth via the oral approach of patients with a history of laparotomy was significantly shorter than that of the patients without a history of laparotomy ([241.6 ± 64.2] cm vs [280.9 ± 40.2] cm, P=0.008). The maximum size of the resected polyps via anus during the second hospitalization was significantly smaller than that during the first hospitalization ([2.25 ± 1.29] cm vs [4.26 ± 3.51] cm, P=0.032). For patients with total enteroscopy, the complete treatment rate was 98% (49/50). For patients without total enteroscopy, all polyps larger than 10 mm in the examined segment of small bowel were resected successfully. Complications occurred in 10 of 129 hospitalizations (delayed bleeding in 4, perforation in 3, and acute pancreatitis in 3). Conclusions: SBE is effective and safe for resection of small bowel polyps in patients with PJS.

The effect of whole body vibration on sensorimotor deficits in people with chronic ankle instability: A systematic review and meta-analysis.

OBJECTIVE: To investigate the effects of whole body vibration on chronic ankle instability-associated sensorimotor deficits in balance, strength, joint position sense and muscle activity. DATA SOURCES: Electronic databases including Cochrane Library, PubMed, Embase, Web of Science, EBSCO, China National Knowledge Infrastructure and WanFang were searched from database inception up to 31 March 2022. METHODS: The risk of bias and methodological quality of included studies were assessed using the Cochrane tool and Physiotherapy Evidence Database (PEDro) scale respectively. Standardized mean difference (SMD) and mean differences (MD) with 95% confidence interval (CI) were calculated using the RevMan 5.3 software. Meta-regression was conducted with Stata 16. RESULTS: Eight studies, with 315 subjects were eventually included in this review with an average PEDro score of 6.1/10. Significant effects of whole body vibration on balance (SMD = 0.61, 95% CI: 0.12 to 1.09, P = 0.01), and on the posterolateral direction (MD = 5.52, 95% CI: 1.02 to 10.01, P = 0.02) and medial direction (MD = 3.90, 95% CI: 0.87 to 6.94, P = 0.01) of the star excursion balance test were found. Whole body vibration significantly improved the peak torque (SMD = 0.36, 95% CI: 0.04 to 0.69, P = 0.03), joint position sense (SMD = 0.60, 95% CI: 0.10 to 1.11, P = 0.02), and muscle activity in tibialis anterior (SMD = 0.46, 95% CI: 0.04 to 0.88, P = 0.03) and gastrocnemius (SMD = 0.68, 95% CI: 0.14 to 1.23, P = 0.01). CONCLUSIONS: The current evidence supports the use of whole body vibration to improve sensorimotor deficits involving balance, strength, joint position sense, and muscle activity in people with chronic ankle instability.

Effects of Pre-exercise Acute Vibration Training on Symptoms of Exercise-Induced Muscle Damage: A Systematic Review and Meta-Analysis.

ABSTRACT: Tan, J, Shi, X, Witchalls, J, Waddington, G, Lun Fu, AC, Wu, S, Tirosh, O, Wu, X, and Han, J. Effects of pre-exercise acute vibration training on symptoms of exercise-induced muscle damage: a systematic review and meta-analysis. J Strength Cond Res 36(8): 2339-2348, 2022-Exercise-induced muscle damage (EIMD) normally occurs after unaccustomed high-intensity eccentric exercises. Symptoms of EIMD include delayed-onset muscle soreness (DOMS), tenderness, stiffness, swelling, reduced strength, and increased creatine kinase (CK) levels in the blood. Vibration training (VT) may be useful as a pre-exercise intervention in attenuating EIMD on the basis of tonic vibration reflex (TVR) through a more efficient distribution of contractile stress over muscle fibers. The objective of this meta-analysis is to examine the effects of acute VT on symptoms of EIMD when performed as the pre-exercise intervention. Randomized controlled trials (RCTs) published in the 8 databases of Cochrane Library, PubMed, Embase, Web of Science, EBSCO, China National Knowledge Infrastructure, Airiti Library and WanFang Data from 1966 (the earliest available time) to January 2019 were searched. A total of 2,324 records were identified and 448 articles were screened with the title and abstract. Two investigators identified eligible studies, extracted data, and assessed the risk of bias independently. Review Manager 5.3 designed by Cochrane was used for the current meta-analysis. Six RCTs involving 180 subjects were included in the analysis. A low-to-moderate methodological quality of the included studies was revealed using the physiotherapy evidence database scale. The results showed that acute VT was superior to the control group for the reduction of DOMS on pain visual analogue scale at 24, 48 hours and pressure pain threshold at 24 hours. In addition, superior effects of acute VT were also found on the indirect markers of muscle damage including CK at 24, 72 hours, and lactate dehydrogenase at 24 hours. The current meta-analysis has collated the evidence to demonstrate that receiving acute VT before unaccustomed high-intensity eccentric exercises may be effective in attenuating markers of muscle damage and the development of DOMS when compared with a control group. The possible mechanisms of this effect could be attributed to an improved synchronization of muscle fiber caused by TVR, which could result in even distribution of exterior loads and eventually attenuate disruptions of muscle fibers. In addition, increased blood flow may also be helpful to prevent accumulation of metabolic substances and attenuate subsequent symptoms of EIMD. Vibration training may be used as a pre-exercise intervention to alleviate symptoms of EIMD caused by unaccustomed high-intensity eccentric exercise. Because of the limited quantity and quality of included studies, more high-quality studies are required to ascertain the effect of VT on symptoms of EIMD.

Mesoporous Silica Carrier-Based Composites for Taste-Masking of Bitter Drug: Fabrication and Palatability Evaluation.

Palatability is one of the most critical characteristics of oral preparations. Therefore, the exploration of new techniques to mask the aversive taste of drugs is in continuous demand. In this study, we fabricated and characterized composites based on mesoporous silica (MPS) that consisted of MPS, a bitter drug, and release regulators. We conducted a palatability evaluation to assess the taste-masking efficacy of the composites. The composites were prepared using the dry impregnation method combined with hot-melt extrusion. Morphology and components distribution in composites were characterized by scanning electron microscopy, confocal laser scanning microscopy, X-ray photoelectron spectroscopy, powder flow properties evaluation, and nitrogen-sorption measurement. The results demonstrated that drugs mainly existed in the inner pore of composites, and release regulators existed in the inner pore and covered the composites' surface. Interactions among the composite components were studied using powder X-ray diffraction, differential scanning calorimetry, and Fourier transform infrared spectroscopy. The drug loaded into the composites was amorphous, and an intermolecular interaction occurred between the drug and the MPS. Taste-masked composites significantly reduced drug release levels under mouth conditions; thus, they prevented the interaction of the dissolved drug with taste receptors and improved palatability. An electronic tongue evaluation and a human taste panel assessment confirmed the better palatability of taste-masked composites. Moreover, the desired drug release behavior can be adjusted by choosing an appropriate release regulator, with stronger hydrophobicity of release regulators resulting in slower drug release. This work has provided new insights into taste-masking strategies for drugs with unpleasant tastes.

[Dual-specificity Phosphatase 1 Suppresses Vascular Smooth Muscle Cell Calcification by Optical Atrophy 1-related Pathway].

Objective To investigate the effect of dual-specificity phosphatase 1/optical atrophy 1 (DUSP1/OPA1) signaling pathway on vascular smooth muscle cell (VSMC) calcification.Methods An in vitro model of VSMC calcification was induced by exposure to ß-glycerophosphate and calcium chloride.VSMC calcification was assessed by Alizarin Red S staining and calcium content by ELISA.Apoptosis was detected by TUNEL.Western blotting was employed to determine the protein levels of DUSP1,OPA1,Runt-related transcription factor 2 (Runx-2),bone morphogenetic protein 2 (BMP-2),and cysteinyl aspartate-specific proteinase-3 (Caspase-3).The effects of DUSP1 overexpression and OPA1 knockdown on cell calcification were investigated.Results Calcium chloride and ß-glycerolphosphate induced VSMC calcification and down-regulated the expression levels of DUSP1 (t=11.951,P<0.001) and OPA1 (t=8.487,P<0.001).DUSP1 overexpression promoted OPA1 expression (t=-8.921,P<0.001),attenuated VSMC calcification,reduced calcium content and apoptosis rate,and down-regulated the expression of Runx-2,BMP-2,and active Caspase-3 (all P<0.001).OPA1 knockdown increased calcium content and apoptosis rate,up-regulated the expression of Runx-2,BMP-2,and active Caspase-3,and promoted VSMC calcification (all P<0.001).Conclusion DUSP1 may inhibit the VSMC calcification through the OPA1 signaling pathway.

Improvement of the activity and SO2 tolerance of Sb-modified Mn/PG catalysts for NH3-SCR at a low temperature.

A series of MnM/palygorskite (PG) (M = La, W, Mo, Sb, Mg) catalysts was prepared by the wetness co-impregnation method for low-temperature selective catalytic reduction (SCR) of NO with NH3. Conversion efficiency followed the order Sb > Mo > La > W > Mg. A combination of various physico-chemical techniques was used to investigate the influence of Sb-modified Mn/PG catalysts. MnSb0.156/PG catalyst showed highest NO conversion at low temperatures in the presence of SO2 which reveals that addition of Sb oxides effectively enhances the SCR activity of catalysts. A SO2 step-wise study showed that MnSb0.156/PG catalyst displays higher durable resistance to SO2 than Mn/PG catalyst, where the sulfating of active phase is greatly inhibited after Sb doping. Scanning electron microscopy and X-ray diffraction results showed that Sb loading enhances the dispersion of Mn oxides on the carrier surface. According to the results of characterization analyses, it is suggested that the main reason for the deactivation of Mn/PG is the formation of manganese sulfates which cause the permanent deactivation of Mn-based catalysts. For Sb-doped Mn/PG catalyst, SOx ad-species formed were mainly combined with SbOx rather than MnOx. This preferential interaction between SbOx and SO2 effectively shields the MnOx as active species from being sulfated by SO2 resulting in the improvement of SO2 tolerance on Sb-added catalyst. Multiple information support that, owing to the addition of Sb, original formed MnOx crystallite has been completely transformed into highly dispersed amorphous phase accounting for higher SCR activity.

Increased expression of hypoxia inducible factor-1 alpha and vascular endothelial growth factor is associated with diabetic gastroparesis.

BACKGROUND: Gastroparesis is a recognized complication of diabetes but its pathogenic mechanism incompletely understood. Our aim was to determine whether HIF-1α and VEGF are secreted from gastric tissue is a fundamental factor that drives diabetic gastroparesis. METHODS: Diabetes was induced in Sprague-Dawley by a single intraperitoneal injection of 65 mg/kg streptozotocin. After 4 and 12 weeks, rats were euthanized for assaying body weight, blood glucose, gastric acid secretion and gastric emptying. Morphologic changes in gastric mucosa were observed by the light microscope. Expression of HIF-1α and VEGF were assessed using immunohistochemistry, RT-PCR and Western blot analyses. RESULTS: Compared with control group, blood glucose were significantly increased and body weight were markedly decreased in streptozotocin-induced diabetes. Gastric emptying was significantly decreased in diabetic rats compared to the control group at different times. The number of parietal cells was obviously decreased, and vacuolated degeneration in diabetic rats. Gastric acid secretion in diabetic group was significantly decreased, and expression of HIF-1α and VEGF were significantly increased in the diabetic group. CONCLUSION: These results indicated that overexpression of HIF-1α and VEGF in the gastric mucosa and played a pivotal role in the progression of diabetic gastroparesis.