We developed the FORCETM platform to overcome limitations of oligonucleotide delivery to muscle and enable their applicability to neuromuscular disorders. The platform consists of an antigen-binding fragment, highly specific for the human transferrin receptor 1 (TfR1), conjugated to an oligonucleotide via a cleavable valine-citrulline linker. Myotonic dystrophy type 1 (DM1) is a neuromuscular disorder caused by expanded CUG triplets in the DMPK RNA, which sequester splicing proteins in the nucleus, lead to spliceopathy, and drive disease progression. Multiple surrogate conjugates were generated to characterize the FORCE platform. DYNE-101 is the conjugate designed to target DMPK and correct spliceopathy for the treatment of DM1. HSALR and TfR1hu/mu;DMSXLTg/Tg mice were used as models of myotonic dystrophy, the latter expresses human TfR1 and a human DMPK RNA with >1,000 CUG repeats. Cynomolgus monkeys were used to determine translatability of DYNE-101 pharmacology to higher species. In HSALR mice, a surrogate FORCE conjugate achieves durable correction of spliceopathy and improves myotonia to a greater extent than unconjugated ASO. In patient-derived myoblasts, DYNE-101 reduces DMPK RNA and nuclear foci, consequently improving spliceopathy. In TfR1hu/mu;DMSXLTg/Tg mice, DYNE-101 reduces mutant DMPK RNA in muscle, thereby correcting splicing. Reduction of DMPK foci in cardiomyocyte nuclei accompanies these effects. Low monthly dosing of DYNE-101 in TfR1hu/mu;DMSXLWT/Tg mice or cynomolgus monkeys leads to a profound reduction of DMPK expression in muscle. These data validate FORCE as a drug delivery platform and support the notion that DM1 may be treatable with low and infrequent dosing of DYNE-101. Oligonucleotides are small pieces of DNA or RNA that can be used to modify expression of genes. Myotonic dystrophy type 1 (DM1) is a severe disorder caused by an abnormal gene that affects multiple organs, including muscle. We developed the FORCE platform to deliver oligonucleotides to muscle. Here we evaluate the impact of this platform on muscle cells from people living with DM1, myotonic dystrophy mouse models, and healthy non-human primates. Our results show that FORCE can deliver oligonucleotides to muscle and provide beneficial effects in animal models of DM1. In the future, FORCE could potentially be used to treat people living with DM1. Weeden et al. use a method called FORCE to deliver oligonucleotides to muscle and evaluate the platform’s utility in models of myotonic dystrophy. An antisense oligonucleotide targeting DMPK is guided to muscle tissue with an antibody fragment and beneficial results are seen in mouse models of disease and cynomolgus monkey.

The full-length cDNA and gDNA sequences of a polyketide synthase gene, termed pks-pa, of Phomopsis asparagi (the fungal pathogen causing the asparagus stem blight), were obtained by RT-PCR, 5'/3'-RACE, PCR and TAIL-PCR. The full-length pks-pa gDNA sequence is 9.336 kb, comprising 5 introns and 6 exons. Sequence alignment revealed a 28-nucleotide difference in polyketide synthase genes between P. asparagi and its closely related species. Phylogenetic analysis indicated that pks-pa is evolutionarily distinct from the polyketide synthases of other fungal species in the same genus. Protein sequence analysis and structural prediction suggested that pks-pa is a hydrophilic protein composed of alpha, beta, turns, coil, alpha amphipathic, beta amphipathic and flexible regions. To confirm the function of pks-pa, a knockout plasmid was used to generate pks-pa knockout transformants in the P. asparagi XT3 isolate. Pathogenic assays showed that loss of pks-pa had no effect on P. asparagi virulence, but significantly reduced pigment production compared with wild-type P. asparagi isolates. These results indicate that pks-pa is required for pigment production, but not P. asparagi virulence.

Purpose/Objective(s) Abemaciclib plus hormone therapy has been the standard adjuvant treatment for high-risk hormone receptor (HR) positive, human epidermal growth factor receptor 2 (HER-2) negative breast cancer and radiotherapy was also recommended for these patients to achieve satisfied local control. Although in vitro studies exhibited favorable tumor cell reduction after concurrent abemaciclib and radiation, the unclear safety precluded the development of clinical trials. To investigate the safety and feasibility of concurrent abemaciclib and postoperative radiotherapy for breast cancer, we proposed this real-world analysis. Materials/Methods We searched for patients who fulfilled the following eligibility criteria, female aged 18-70 years old, pathologically diagnosed as breast cancer, with stage II-III, received breast conserving surgery or mastectomy followed by standard chemotherapy, received concurrent radiotherapy and abemaciclib in our institution between December 2021 to December 2022. Those who with breast sarcoma or incomplete medical records were excluded. Data regarding the clinicopathological characteristics, treatment details, adverse events (AEs) during radiation and within 12 months after the radiation were collected. AEs were reassessed according to National Cancer Institute common terminology criteria for adverse events, version 5.0. Results From December,2021 to December,2022, 10 patients fulfilled the criteria and were analyzed in this study. The median age was 51.5 years old (ranged 36-58 years old). Seven patients (70%) were with disease of pathological or yp stage III. Neoadjuvant chemotherapy was administrated to 5 patients. Nine patients received chest wall irradiation and the other 1 patient received whole breast irradiation plus tumor bed boost. Regional lymph node irradiation, including superior clavicular and subclaviscular were delivered to all the ten patients. Four patients received internal mammary lymph node irradiation. All of the patients received volumetric modulated arc therapy. Conventional fractionation (2Gy per fraction) and hypofractionation radiotherapy (2.9Gy per fraction), was delivered to 2 and 8 patients. Abemaciclib was prescribed for 150mg twice daily. All of the patients finished the radiation without dose reduction or suspending. Abemaciclib dose reduction to 100mg twice daily was observed in 1 patient. No Grade 4-5 AE was not observed. Totally, Grade 3 AE were recorded in 1 patient, who was with Grade 3 leukopenia and recovered after treatment. Grade 2 interstitial pneumonia (both in radiation field and after radiation field) were recorded in 1 patient, who was diagnosed at 6 months after radiation (more likely attribute to abemaciclib) and recovered after intervention. The most common AE was diarrhea (Grade 1, 40% and Grade 2, 60%). Conclusion Concurrent abemaciclib and postoperative radiotherapy is safe with tolerable and controllable toxicities. Prospective studies with large samples were warranted.

Purpose/Objective(s) Both capecitabine and radiotherapy have been widely used for patients with breast cancer. Although concurrent capecitabine and radiotherapy showed favorable efficacy and tolerable toxicities in patients with gastrointestinal carcinoma, there is still a paucity of real-world evidence on the safety of the concurrent treatment in breast cancer. The aim of this study is to investigate the toxicity of concurrent radiotherapy with capecitabine in breast cancer patients. Materials/Methods Patients pathologically diagnosed as breast cancer, aged 18-70 years old, received concurrent radiotherapy and capecitabine between January 2017 and December 2020, radiation target volume involved chest wall or whole breast with and without lymph node regions, with complete medical record were selected and analyzed. Patients’ characteristics, tumor information, treatment details including irradiation dosimetric index, systemic regimens doses were assembled. Both acute and chronic adverse events (AEs) were evaluated by a senior radiation oncologist, a senior medical oncologist and a radiologist, according to National Cancer Institute common terminology criteria for adverse events, version 5.0. Results Totally, fifty-three patients were analyzed. Baseline characteristics are shown in Table I. Four of them received concurrent chemoradiotherapy after local recurrence. Hypofractionation radiation, with total dose of 43.5Gy-49.5Gy, 2.9Gy per fraction, were delivered to 36 patients and the remaining patients received conventional fractionation radiotherapy, with dose of 50-62.5 Gy, 1.8-2Gy per fraction. Thirty-nine patients (73.6%) were treated with concurrent capecitabine at a dose of 1000mg/m2 bid d1-14/q3w, and the remaining 14 patients (26.4%) were treated with a low-dose maintenance therapy at 500mg tid. The median follow-up time was 26.5 months. The most common side effects were dermatitis (N = 45) and leukopenia (N = 33) at any grade. Followed by radiation esophagitis (N = 17), fatigue (N = 16) and hand-foot syndrome (N = 15). The most common G3 side effects were radiation induced dermatitis (N = 11) and hand-foot syndrome (N = 4). All of them were cured after intervention. All of the patients finished the whole course of radiotherapy. None but 1 patient discontinued concurrent medication because of gastrointestinal reaction. No grade 4-5 AE was observed. Conclusion In general, concurrent radiotherapy and capecitabine in breast cancer were safe with acceptable and controllable toxicities. Dermatitis should be paid more attention in clinical works.

The prognosis of microinvasive breast cancer (MIBC) is between ductal carcinoma in situ and T1aN0 stage tumors. We conducted this real-world study to explore the prognosis of human epidermal factor receptor 2 (HER2)-positive MIBC, as well as to find out whether chemotherapy and anti-HER2 therapy could improve outcome in this population. Patients who received radical surgery and were diagnosed with lymph node-negative HER2-positive MIBC at the National Cancer Center in China from January 2010 to December 2019 were consecutively enrolled. The invasive components were confirmed HER2-positive by pathologists from our center. The exclusion criteria included: 1) distant metastasis before surgery; 2) HER2 status of the invasive components was unknown. Clinicopathologic characteristics and follow-up outcome data were collected. A total of 121 patients were included. The median age was 51 years old, and the median follow-up time was 68.3 months. Twenty-four patients (19.8%) received adjuvant chemotherapy (CT), of which 7 patients received CT combined with trastuzumab. Among these 24 patients, 17 patients were aged ≤ 50, and 20 patients were estrogen receptor (ER) negative. In total, 5 patients experienced recurrence (2 in situ breast recurrence, 3 distant metastasis), all within 3 years after surgery. The 3-year disease free survival (DFS) rate and 5-year DFS rate were all 95.9%. Univariate analysis showed that patients aged ≤ 50 might have worse outcomes than those aged > 50 (5-year DFS rate 92.5% versus 98.5%, p=0.098). Using propensity score matching, patients who received CT with or without trastuzumab were matched with those who did not receive CT in a 1:2 ratio based on age and ER status. Patients who received CT with or without trastuzumab showed a trend of increase in 5-year DFS rate compared to those who did not receive CT (100% vs 89.6%, p=0.106). HER2-positive MIBC had a relatively good prognosis. For patients with risk factors of relapse, CT and trastuzumab might decrease recurrence, so adjuvant CT and anti-HER2 therapy could be considered in this population.

Stephania tetrandra S. Moore belongs to the family Menispermaceae and is a Chinese medicinal plant widely distributed in tropical and subtropical regions of Asia and Africa. The root can be used for a variety of treatments (Jiang et al. 2020). In August 2021, leaf spot symptoms were observed on S. tetrandra cultivated in Jiangxi (114.456E, 27.379N, southern China). The disease symptoms included a slight constriction of the leaves, with irregularly shaped brown to black spots with well-defined borders. Severely affected leaves were shed by the plant. In order to determine the cause, symptomatic leaves were surface-disinfested with 0.6% NaOCl for 2 min, and rinsed twice in sterile water, then incubated on moist paper towels at 26°C in the dark for 2 days. Cream-colored sporodochia were observed within the leaf spots, turning dark green to black within 16 hours. A slow-growing white fungus was isolated from 95% of the samples (n = 30) on PDA. Dark green sporodochia emerged after 7 to 10 days of incubation, and released tip-end oval, non-septate, hyaline conidia measuring 6.7 to 8.5 μm (mean 7.5 μm, n = 50) by 2.0 to 3.3 μm (mean 2.7 μm, n = 50). Concentric rings were interspersed with sporodochia on the continually incubated mycelium. The morphological characteristics of the isolates matched the description of Albifimbria (Lombard et al. 2016). Nucleotide sequences, amplified from isolate FJL5C using primers of the internal transcribed spacer (ITS) (White et al. 1990), calmodulin (cmdA; Carbone and Kohn 1999), and RNA polymerase II second largest subunit (rpb2; O'Donnell et al. 2007), were deposited in GenBank under accession numbers OM317911, OM386815, and OM386816. A BLASTn analysis of the sequences showed 100% identity with the type strain CBS 328.52 (Lombard et al. 2016) of Albifimbria verrucaria (syn. Myrothecium verrucaria) for ITS, and 99% for cmdA and rpb2 (KU845893, KU845875, and KU845931, respectively). A phylogenetic tree generated using the three sequences showed that the isolate from S. tetrandra grouped with the A. verrucaria isolates, but away from other species of Albifimbria. These results together with the lack of a pale luteus exudate produced by A. viridis (Lombard et al. 2016) implied that the isolate was A. verrucaria. The culture was deposited in Guangdong Microbial Culture Collection Center (GDMCC 3.716). To verify pathogenicity, conidial suspension (106 conidia/mL in 0.05% Tween 20 solution) was sprayed onto six healthy plants. Six other plants sprayed with the Tween 20 solution alone served as controls. All plants were incubated in the dark at 26°C and 95% humidity for 30 hours, then transferred to a greenhouse at 26°C and 12 hours of illumination per day for 2 to 3 days. Inoculated leaves developed similar symptoms to those described above, whereas control plants remained healthy. The same pathogen was isolated from the diseased leaves, with the same morphological and molecular traits as those from the field plants. This experiment fulfilled Koch's postulates and confirmed that A. verrucaria causes leaf spots on S. tetrandra. This pathogen has been reported to cause disease in a wide range of weeds, legumes, and crop plants (Herman et al. 2020). To our knowledge, this is the first report of A. verrucaria causing leaf spots on S. tetrandra in natural or controlled environments. The disease can seriously threaten S. tetrandra on growth and yield loss.

TPS9131 Background: Preclinical and clinical evidence has demonstrated that the dual blockade of the EGFR and VEGF pathways is a viable strategy in the EGFR-mutated advanced NSCLC population. Anlotinib is an oral multi-targeted tyrosine kinase inhibitor (TKI) that effectively inhibits VEGFRs, FGFRs, PDGFRs, c-kit and MET. It has been proved to be safe and effective in advanced lung cancer after second-line standard chemotherapy failure. A cohort study of Anlotinib plus Erlotinib has shown a favorable safety profile and promising antitumor activity with an objective response rate (ORR)92.6%. This phase III study aims to evaluate the efficacy and safety of Anlotinib or placebo plus Gefitinib in patients(pts) with untreated EGFR-mutated metastatic NSCLC. Methods: Eligible pts were aged 18̃75 years old, had stage IIIB or IV NSCLC, with an EGFR 19del or 21L858R mutation, an ECOG PS of 0 or 1, measurable lesion according to RECIST v1.1 and adequate organ function. We randomly assigned eligible pts in a 1:1 ratio to receive oral Gefitinib (250 mg QD) plus either Anlotinib (12 mg QD from day 1 to 14 of a 21-day cycle) or matching placebo until progressive disease or unacceptable toxicity. Randomization was done by an interactive web response system with a computer-generated sequence and stratified by EGFR mutation status, gender, ECOG PS and pathological type. The primary endpoint is progression-free survival(PFS). Secondary endpoints include overall survival, ORR, disease control rate, time to progression, duration of response, quality of life and the safety profile. The peripheral blood of the pts will be detected three times by polygenic detection to monitor the resistance mechanism (before treatment, during the first evaluation, during tumor progression, each time 10ml peripheral blood). Independent Data Monitoring Committee and Independent Review Committee will be used in this study. According to previous report (Erlotinib plus Bevacizumab vs. Erlotinib alone: 16.0 vs. 9.7 mos, HR 0.54, Lancet Oncol, 15(11):1236-1244), the sample size was determined based on a median PFS of 15 months for the Anlotinib + Gefitinib group and median PFS of 10 months for the Placebo + Gefitinib group. To achieve 80% power at a two-sided α = 0.05 and an anticipated dropout rate of 20%, 310 patients (with 192 events required for the analyses) were needed. In total, 310 patients will be enrolled in this trial at 16 sites in China. From April 2019, 224 patients have been enrolled. Clinical trial information: NCT04028778.

Stephania tetrandra S. Moore is a perennial liana and is widely cultivated in southern China for traditional Chinese medicine as a diuretic, anti-inflammatory, and antirheumatic treatment (Jiang et al. 2020). In August 2021, it was observed that a severe stem rot disease affected St. tetrandra cultivated in Anfu, Jiangxi province, China (114°27'26" E, 27°22'46" N). The disease symptoms included constriction and rot at the base of the stem, and covered with a layer of white mycelia. The plants above-ground finally wilted and dried with a disease incidence ranging from 8% to 16%. Lots of dried plants formed withered patches of field. Sections (1.0~2.0 cm) from browning stem tissues were surface-disinfected with 75% ethanol for 15 s, followed by 60 s in 4% NaClO, rinsed twice in sterile water, dried on sterilized filter paper, placed on potato dextrose agar (PDA), and incubated at 26°C in the dark for 3 days. A white rhizomorphic fungal mycelium, that is similar to the mycelium of strain FJSR0 on the surface of an infected plant in the field, was isolated from the cultured tissues with 67% frequency. When incubated on PDA, white and fluffy mycelia with even margins and a slight halo formed. Mycelia-produced clamp connections were observed. Colonies grew quickly and covered the dish (diameter: 9 cm) in 5 or 6 days. After that, sclerotia were initially white, then turned yellow, and chestnut brown at maturity. Spherical and subspherical sclerotia were observed after 8 days, with each plate containing 448 to 634 sclerotia (0.8 to 1.4 mm diameter; mean = 0.94 mm; n = 50). On the basis of morphology, the pathogen was similar to Sclerotium rolfsii Sacc. [teleomorph: Athelia rolfsii (Curzi) Tu & Kimbrough] (Sun et al. 2020; Ling et al. 2021). For molecular confirmation, the internal transcribed spacer (ITS) region with approximately 680 bp was amplified from strains FJRS0 and FJRS1 using primers ITS1/ITS4 (White et al. 1990). Two distinct types (different in one SNP and one 1-bp InDel) of ITS sequences were obtained from each isolate, and all isolates contain the two types (FJSR0: ON972516, ON972517; FJSR1: ON972520, ON972518). BLAST analysis of each type found that the hits, with identities >99%, are A. rolfsii except for two Sc. delphinii sequences (GU567775.1 and MK073010.1). Phylogenetic analysis placed strains FJSR0 and FJSR1 in the same clade as Sc. rolfsii but away from Sc. delphinii based on the previous method (Sun et al. 2021). Both morphological and molecular characteristics confirmed that the strains were Sc. rolfsii. For pathogenicity tests, PDA plugs (8 mm in diameter) covered with 5-day-old fungal mycelium were inoculated at the stem bases of three healthy St. tetrandra seedings and incubated at 26℃ and relative humidity of 80%. On the fifth day, inoculated plants were wilting. The infected stem bases turned brown to black and constricted as previously observed in the field. Some leaves, infected by the mycelium expanded from the PDA plugs, developed an orange and irregular spot. Sclerotia were observed 20 days post inoculation. In contrast, the leaves and stems of non-inoculated control plants remained symptomless. Pathogenicity tests were repeated three times. The fungus was reisolated consistently from each symptomatic tissue, thus completing Koch's postulates. Although Sc. rolfsii has been previously reported to cause a southern blight symptoms on vegetables, ornamentals, grass, and medicinal and leguminous crops (Sun et al. 2020; Ling et al. 2021), this is the first report of Sc. rolfsii causing similar symptoms of southern blight on St. tetrandra in China.

Abstract Background CCAAT/Enhancer Binding Protein D (CEBPD), a pleiotropic glucocorticoid-responsive transcription factor, modulates inflammatory responses. Of relevance to asthma, expression of CEBPD in airway smooth muscle (ASM) increases with glucocorticoid exposure. We sought to characterize CEBPD-mediated transcriptomic responses to glucocorticoid exposure in ASM by measuring changes observed after knockdown of CEBPD and its impact on asthma-related ASM function. Methods Primary ASM cells derived from four donors were transfected with CEBPD or non-targeting (NT) siRNA and exposed to vehicle control, budesonide (100 nM, 18 h), TNFα (10 ng/ml, 18 h), or both budesonide and TNFα. Subsequently, RNA-Seq was used to measure gene expression levels, and pairwise differential expression results were obtained for exposures versus vehicle and knockdown versus control conditions. Weighted gene co-expression analysis was performed to identify groups of genes with similar expression patterns across the various experimental conditions (i.e., CEBPD knockdown status, exposures). Results CEBPD knockdown altered expression of 3037 genes under at least one exposure (q-value < 0.05). Co-expression analysis identified sets of 197, 152 and 290 genes that were correlated with CEBPD knockdown status, TNFα exposure status, and both, respectively. JAK-STAT signaling pathway genes, including IL6R and SOCS3, were among those influenced by both TNFα and CEBPD knockdown. Immunoblot assays revealed that budesonide-induced IL-6R protein expression and augmented IL-6-induced STAT3 phosphorylation levels were attenuated by CEBPD knockdown in ASM. Conclusions CEBPD modulates glucocorticoid responses in ASM, in part via modulation of IL-6 receptor signaling.

Objective:To explore the dynamic changes of Distress Thermometer scores and the relationship between psychological distress and quality of life in Chinese early breast cancer patients during chemotherapy.Methods:This prospective study enrolled 110 Chinese postoperative early breast cancer patients between March 2019 and December 2019. The psychological distress and quality of life (QOL) of patients were assessed by using the psychological distress management screening tool and the patient quality of life scale. Logistic regression model was used to analyze the influencing factors of psychological distress degree. The correlation between distress thermometer (DT) score changes and quality of life was analyzed by Pearson correlation analysis.Results:In total, 96 valid cases were analyzed. Before chemotherapy, 47 cases (49.0%) had DT score ≥4 points. After 2 cycles of chemotherapy, 40 cases (41.7%) had DT score ≥4 points. Thirty-four patients (35.4%) had DT score ≥4 points after chemotherapy. The DT score after chemotherapy was lower than that before chemotherapy and after 2 cycles of chemotherapy. Univariate analysis showed that income level and pathological stage were still significant related to the detection of DT score ≥4 points after chemotherapy ( P<0.05). The changes of DT scores before and after chemotherapy were negatively correlated with the changes of quality of life ( r=-0.298, P=0.003). Conclusions:The detection rate of psychological distress in patients with early breast cancer during chemotherapy showed a decreasing trend. Income level and tumor stage are significant factors affecting the psychological distress of patients. There is a significant correlation between the psychological distress and the quality of life during chemotherapy. We should pay attention to the evaluation and monitoring state of psychological distress of patients during chemotherapy.