Small RNAs (sRNAs), the key components of RNA interference (RNAi) or RNA silencing, can mediate cell-autonomous gene silencing and function as signaling molecules across species. Microbe-induced gene silencing (MIGS), which is based on interspecies RNAi, is an effective approach for controlling fungal diseases in crops. The enolase gene VdEno is essential for the growth and development of the fungal pathogen Verticillium dahliae, which causes cotton Verticillium wilt. In this study, we engineered Trichoderma harzianum (Th) to express the double-stranded RNA (dsRNA) targeting VdEno. The engineered strain Th-VdEnoi successfully generated VdEno-specific small interfering RNA (siVdEno). We further confirmed that Th-VdEnoi effectively induced VdEno silencing at the translational level. The results of crop protection assays revealed that the cotton plants co-inoculated with V. dahliae (strain V592) and Th-VdEnoi presented significantly reduced disease severity and lower fungal biomass in their roots than the control plants inoculated with V. dahliae alone or with V. dahliae and Th-GFPi (a control strain expressing GFP-targeting dsRNA). Collectively, our findings demonstrate that VdEno is an effective target for controlling cotton Verticillium wilt and confirm that MIGS is a promising strategy for managing soil-borne fungal pathogens in crops. MIGS provides strong technical support for reducing the application of conventional chemical pesticides, developing eco-friendly biopesticides, and facilitating the sustainable development of agriculture.
Gossypium/microbiology* ; Plant Diseases/prevention & control* ; Gene Silencing ; Ascomycota/genetics* ; RNA Interference ; RNA, Double-Stranded/genetics* ; Hypocreales/genetics* ; RNA, Small Interfering/genetics* ; Verticillium/genetics* ; Fungal Proteins/genetics*

Objective:To investigate the efficacy and safety of combining venetoclax (VEN) with hypomethylated drugs (HMA) in the treatment of higher-risk (IPSS-R score >3.5) myelodysplastic syndromes (MDS) .Methods:From March 2021 to December 2022, forty-five MDS patients with intermediate and high risk were treated with VEN in combination with HMAs. Clinical data were collected and analyzed retrospectively, including gender, age, MDS subtype, IPSS-R score, treatment regimen, and efficacy, etc. Kaplan-Meier method and Cox regression model were used to analyze univariate and multivariate of survival prognosis.Results:①Forty-five patients with MDS, including ninety-one percent were classified as high or very high risk. According to the 2023 consensus proposal for revised International Working Group response criteria for higher-risk MDS, the overall response rate (ORR) was 62.2% (28/45), with the complete response rate (CR) was 33.3% (15/45). For twenty-five na?ve MDS, the ORR was 68% (17/25) and the CR rate was 32% (8/25). In nonfirst-line patients, the ORR and CR were 55% (11/20) and 35% (7/20) respectively. The median cycle to best response was 1 (1-4). ②With a median followup of 189 days, the median overall survival (OS) time was 499 (95% confidence interval, 287-711) days, and most patients died from disease progression. Responders had a significantly better median OS time than nonresponders (499 days vs 228 days, P<0.001). Multifactor analysis revealed that IPSS-R score and response to treatment were independent prognostic factors for OS; the presence of SETBP1 gene mutations was associated with a longer hospital stay (51.5 days vs 27 days, P=0.017) . Conclusions:There is clinical benefit of venetoclax in combination with hypomethylated agents in patients with higher-risk MDS, but adverse events such as severe hypocytopenia during treatment should be avoided.

Objective:To explore the mechanism of peritoneal dialysis solution (PDS)-induced peritoneal microinflammation through activation of ceramide (CER) in peritoneal dialysis model mice.Methods:Thirty 5-week-old male C57BL/6 mice weighing about 22 g were used to set up peritoneal dialysis models, and then were randomly divided into 4 groups: sham operation group (1.5 ml sterilized water, n=7), high glucose-PDS group (1.5 ml 4.25% PDS, n=8), high glucose-PDS+ acid sphingomyelinase (ASMase) inhibitor desipramine (DES) group (1.5 ml sterilized water+10 mg/kg DES, n=8), high glucose-PDS+Src kinase inhibitor PP2 group (1.5 ml sterilized water +1 mg/kg PP2, n=7), with intraperitoneal injection once a day. After 28 days, the mice were sacrificed to retain peritoneal tissues. HE staining and Masson staining were used to observe the histological changes of peritoneum. Immunohistochemistry was used to detect the Toll-like receptor 4 (TLR4) and macrophages. High performance liquid chromatography, liquid chromatography/mass spectrometry and immunofluorescence were used to detect the expression of ASMase and CER. Real-time quantitative PCR was used to detect the mRNA levels of c-Src, p-Src, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Western blotting was used to detect the protein levels of c-Src, and p-Src. Enzyme-linked immunosorbent assay was used to detect the serum C reactive protein (CRP), IL-6 and TNF-α. Results:(1) High glucose-PDS led to peritoneal hyperplasia, collagen deposition and fibrosis in the peritoneal dialysis mice, indicating successful modeling. Compared with high glucose-PDS group, peritoneal hyperplasia, collagen deposition and fibrosis of mice treated with DES and PP2 were significantly improved (all P<0.05). (2) Compared with sham operation group, ASMase activation and CER level of peritoneal tissues were significantly higher in high glucose-PDS group, and DES could significantly inhibit activated ASMase and increased CER expression caused by high glucose-PDS (both P<0.05). PP2 had no significant effect on ASMase activation and CER level (both P>0.05). (3) Compared with sham operation group, there were more TLR4 and macrophage positive staining cells in peritoneal tissues in high glucose-PDS group, and the mRNA expression levels of IL-6 and TNF-α in peritoneal tissues and serum CRP, IL-6 and TNF-α were higher (all P<0.05). DES and PP2 could significantly inhibit the increased TLR4, macrophages and related inflammatory factors induced by high glucose-PDS (all P<0.05). (4) Compared with sham operation group, c-Src and p-Src mRNA and protein expression levels of peritoneal tissues in high glucose-PDS group were significantly higher (all P<0.05). PP2 significantly inhibited the increased p-Src mRNA and protein levels caused by high glucose-PDS (both P<0.05), but had no significant effect on the mRNA and protein expression levels of c-Src (both P>0.05). DES had no significant effect on the mRNA and protein expression levels of c-Src and p-Src (all P>0.05). Conclusions:High glucose-PDS may enhance the expression of CER through stimulating the activity of ASMase, phosphorylate Src, activate TLR4 and induce inflammatory damage of peritoneum in peritoneal dialysis model mice.

Acquired immune deficiency syndrome is an infectious disease with high mortality caused by human immunodefi-ciency virus(HIV).Even after antiretroviral therapy,the virus reservoir formed by the provirus integrated in the host genome can evade host immune surveillance and clearance.The existing methods of HIV reservoir detection mainly include cell culture induced virus growth assay in vitro and direct detection of the provirus genome based on PCR technology.Understanding and measuring accurately of HIV reservoir can provide reliable technical basis for HIV treatment research.This article summarizes and analyzes various detection methods of HIV reservoirs in recent years and their advantages and disadvantages,in order to select the suitable method for different detection objects of HIV reservior,and give appropriate suggestions,to provide technical support and theoretical guidance for further research on acquired immune deficiency syndrome.

Objective To investigate the effect and mechanism of macrophage colony stimulating factor(M-CSF)on myocardial function after acute myocardial infarction(AMI)in mice by regulating cardiac macrophages.Methods Fifty C57mice were randomly di-vided into Sham Group(n=10,sham operation group),MI Group(n=20,AMI model was prepared,and normal saline was injected in-traderitoneally)and MM Group { n=20,AMI model was prepared,and M-CSF reagent[500μg/(kg·d)]was injected intraderitoneal-ly}.At the end of the experiment,after completing a cardiac ultrasound,the myocardial tissue was collected,interleukin-4(IL-4),interleukin-6(IL-6),monocyte ehemoattractant protein-1(MCP-1),tumor necrosis factor-α(TNF-α),interleukin-10(IL-10),interferon-α(IFN-α),atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),Collagen Ⅰ and Collagen Ⅲ were detected by enzyme-linked immunosorbent assay(ELISA),the apoptosis-related proteins Bax,C-caspase-3,caspase-3,nuclear transcription facter(NF)-κB p65,transduction and activator of transcription 3(STAT3),transduction and activator of transcription 6(STAT6)were detected by Western blot assay,and the neovascularization markers in MI peripheral area were determined by immunohis-tochemistry,the expression levels of M1-type macrophages and M2-type macrophages were detected by flow cytometry.Results The left ventricular size and left ventricular ejection fraction(LVEF)were significantly improved in MM Group when compared with the MI Group,and the indexex of myocardial inflammation,hypertrophy,apoptosis,and fibrosis decreased significantly(P＜0.05).The expres-sion of neovascularization mardker CD31 was significantly up-regulated in MM Group.The expression of M2macrophage in MM group was significantly higher than when compared with the MI group,while the level of M1 macrophage was lower(P＜0.05).The expression of NF-κB p65 in MI group was statistically higher than that in the Sham group and MM group,while the levels of STAT3 and STAT6 were higher in the MM group than in the MI group(P＜0.05).Conclusion M-CSF can inhibit inflammatory response,reduce myo-cardial fibrosis,hypertrophy and apoptosis,promote angiogenesis,inhibit M1-type macrophages and up-regulate the expression of M2-type macrophages to promote myocardial tissue repair and improve ventricular structural remodeling,in which NF-κB/STAT signa-ling pathway plays an important role.

Objective:To explore the differences in clinical characteristics and treatment outcomes between patients with type A and type B alcohol dependence, and to find the independent risk factors of relapse.Methods:Alcohol-dependent male patients attending the Addiction Medicine Center of Beijing Huilongguan Hospital from January 2018 to December 2020 were selected for the study and divided into type A alcohol-dependent group ( n=77) and type B alcohol-dependent group ( n=87). All patients were given acute detoxification treatment and were followed up after treatment on relapse to drinking. Differences in demographic and clinical data were compared between the two groups, and differences in treatment outcomes between the two groups at different time points over 3 months were compared. Patients were divided into relapse group and non-relapse group according to whether they drank again after 3 months. Logistic regression model was established to screen the risk factors of relapse of alcohol-dependent patients by SPSS 25.0 software. Results:There was no significant difference between the two types of patients in years of education, marital status, smoking status and working status(all P>0.05), but the proportion of co-residents( χ2=5.69, P=0.017) and the proportion of positive family history of alcoholism were significant difference between the two type of patients( χ2=13.32, P<0.001). There were statistically significant differences between the two types of patients in the onset time( t=-7.28, P<0.001), the first drinking age( t=-2.36, P=0.020), the proportion of drinking in the morning( χ2=7.83, P=0.005), psychotic symptoms( χ2=4.31, P=0.038), convulsions after withdrawal( χ2=5.30, P=0.021), and alcohol use disorder identification test(AUDIT) score( t=4.30, P<0.001). At the 4th and 8th weekend of the follow-up, there were statistically significant differences in drinking frequency(0(0, 3), 0(0, 0), Z=-4.13, P<0.001; 3(0, 3), 0(0, 3), Z=-4.42, P<0.001) and relapse rate (40(45.98%), 9(11.69%), χ2=22.92, P<0.001; 61(70.11%), 24(31.17%), χ2=24.82, P<0.001) between the two types of alcohol dependence patients after drinking again. After 12-week follow-up, there were statistically significant differences between the two types of alcohol-dependent patients in the interval of first drinking(20(7, 30)d, 88(38, 90)d, Z=-7.83, P<0.001), the cumulative duration of abstinence(4(0, 8)weeks, 12(4, 12)weeks, Z=-5.13, P<0.001), the cumulative rate of abstinence(71(81.60%), 25(32.47%), χ2=40.62, P<0.001), the frequency of drinking after abstinence(3(3, 3), 0(0, 3), Z=-5.54, P<0.001), and the reduction of daily average alcohol consumption( t=3.36, P<0.001). Logistic regression model showed that type B alcohol dependence ( OR=3.121, P=0.03, 95% CI: 1.12-8.72) and AUDIT score ( OR=1.498, P<0.01, 95% CI: 1.29-1.74) were the risk factors for relapse of alcohol-dependent patients. Conclusions:Patients with type A and type B alcohol dependence have obvious differences in clinical characteristics and treatment outcomes, and type B alcohol dependence is independent risk factor for relapse to drinking in alcohol-dependent patients, which validate the rationality and necessity of alcohol dependence subtypes.

Objective:To explore the risk factors in leukemia transformation (LT) in those with myelodysplastic syndromes (MDS) .Methods:From January 2012 to December 2020,data on 320 patients with newly diagnosed primary MDS were gathered from the MDS center. The clinical features and molecular characteristics are explored. Additionally, a retrospective analysis of risk factors for the development of acute leukemia from MDS was done.Results:The median follow-up was13.6 (0.4-107.3) months. 23.4% (75/320) of the MDS patients had LT group. Significant differences between the LT group and non-LT group can be seen in age ( P<0.001) , bone marrow blast percentage ( P<0.001) , bone marrow fibrosis ( P=0.046) , WHO classification ( P<0.001) , IPSS-R ( P<0.001) and IPSS-R karyotype group ( P=0.001) . The median number of mutation of LT group was 1 (1, 3) , that in non-LT group was 1 (0, 2) ,which had a statistical difference ( P=0.003) .At the time of the initial diagnosis of MDS, the LT group had higher rates of the TP53 mutation ( P=0.034) , DNMT3A mutation ( P=0.026) , NRAS mutation ( P=0.027) and NPM1 mutation ( P=0.017) . Compared with the mutations at first diagnosis and LT of six patients, the number of mutations increased and the variant allele frequencies (VAF) increased significantly in LT patients. Higher bone marrow blast percentage (Refer to <5% , 5% -10% : HR=4.587, 95% CI 2.214 to 9.504, P<0.001, >10% : HR=9.352, 95% CI 4.049 to 21.600, P<0.001) , IPSS-R cytogenetic risk groups ( HR=2.603, 95% CI 1.229-5.511, P=0.012) , DNMT3A mutation ( HR=4.507, 95% CI 1.889-10.753, P=0.001) , and NPM1 mutation ( HR=3.341, 95% CI 1.164-9.591, P=0.025) were all independently associated with LT in MDS patients, according to results of multivariate Cox regression. Conclusion:Bone marrow blast percentage, IPSS-R cytogenetic risk groups, DNMT3A mutation, and NPM1 mutation are independent risk factors in LT for MDS patients.

Adipose-derived mesenchymal stem cells (ASCs) are pluripotent adult mesenchymal stem cells that are expected to be ideal seed cells for bone tissue engineering(BTE) due to their biosecurity, abundance, and easy access. However, in comparison with bone marrow mesenchymal stem cells, ASCs possess a relatively limited osteogenic capacity and often require further induction. Advances on strategies for promoting osteogenic differentiation of ASCs are reviewed from following aspects: optimization of scaffolds, addition of bioactive factors or drugs, non-coding RNA regulation, and physical stimulation, so as to provide references for the use of ASCs in BTE.

Adipose-derived mesenchymal stem cells (ASCs) are pluripotent adult mesenchymal stem cells that are expected to be ideal seed cells for bone tissue engineering(BTE) due to their biosecurity, abundance, and easy access. However, in comparison with bone marrow mesenchymal stem cells, ASCs possess a relatively limited osteogenic capacity and often require further induction. Advances on strategies for promoting osteogenic differentiation of ASCs are reviewed from following aspects: optimization of scaffolds, addition of bioactive factors or drugs, non-coding RNA regulation, and physical stimulation, so as to provide references for the use of ASCs in BTE.

OBJECTIVE: To explore the clinical and genetic characteristics of a child with frontometaphyseal dysplasia 1 (FMD1) due to variant of FLNA gene. METHODS: Clinical phenotype of the patient was analyzed. Whole exome sequencing (WES) was carried out to detect pathogenic genetic variants. Sanger sequencing was used to verify the result in his parents. RESULTS: The 2-year-and-9-month-old boy presented with facial dysmorphism (supraorbital hyperostosis, down-slanting palpebral fissure and ocular hypertelorism), skeletal deformities (bowed lower limbs, right genu valgum, left genu varus, slight deformity of index and middle fingers, and flexion contracture of little fingers). He also had limited left elbow movement. High-throughput sequencing revealed that he has carried a de novo heterogeneous c.3527G>A (p.Gly1176Glu) missense variant of the FLNA gene. The same variant was found in neither parent. CONCLUSION The clinical manifestations of FMD1 such as joint contracture and bone dysplasia can occur in infancy and deteriorate with age, and require long-term follow-up and treatment. Above finding has expanded the spectrum of FLNA gene variants.
Child ; Filamins/genetics* ; Forehead/abnormalities* ; Humans ; Infant ; Male ; Osteochondrodysplasias/genetics* ; Phenotype ; Whole Exome Sequencing