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In the present study, a finite element impact model was developed and analyzed using commercial FEM code ANSYS® and then validated via a drop-weight impact experiment. Moreover, double-impactor impact models were designed and developed with different impact distribution and locations of two impactors to compare impact properties. A total of 18 impact scenarios comprised of asymmetric and symmetric types were performed. The effect of impact location on the impact resistance force and duration time was investigated: the closer the impact point is to the fabric center, the longer the impact duration time. In addition, the effect of impact location on impactor failure deflection was also investigated and it was concluded that regardless of the symmetric or asymmetric impact scenario, the smaller the average distance between the impact location of the two impactors from the fixed boundary, the smaller the overall average failure deflection that occurs. The relevance of impact location and fabric energy absorption capacity was also identified. Furthermore, the effect of impact location on fabric stress distribution and transverse deformation and of the variation of the impact velocity on fabric impact behaviors were investigated. These findings will provide important guidance for engineering soft body armor and composite materials.

In China, the invasion of Spartina alterniflora is an important driver for the decrease of mangrove area and ecological service functions related to this habitat. In the past few decades, S. alterniflora clearing and mangrove restoration projects have mainly focused on the areas where it is already changed but ignored the potential distribution areas. This study suggested that implementation of mangrove protection prior to the areas with the threat of S. alterniflora invasion could greatly improve protection efficiency and save costs. Thus, using Maximum Entropy Modeling (MaxEnt), we estimated the potential spatial distribution of both mangroves and S. alterniflora in China, considering the current distribution data, topographical, sediments, sea surface temperature and bioclimatic variables. What’s more, we identified and calculated the potential distributed areas in each province. We aimed to explore (i) the key factors determining the distribution of mangrove and Spartina alterniflora along the coastline and (ii) the hotspots of their competitive occurrence, including S. alterniflora invasion areas and mangroves degradation areas, in order to support mangrove conservation. The model showed that the distance to the coastline and the topography play important roles in the distribution of S. alterniflora, while mangroves were more sensitive to the range of the annual sea surface temperature. Our results furthermore confirm that S. alterniflora has a wider potential distribution area (~10,585 km2) than mangroves (~9124 km2) at the coastline of China; and predict the provinces Zhangzhou, Quanzhou, Zhanjiang, Beihai and Wenzhou as hotspots for the competition between mangroves and S. alterniflora. We propose that priority should be given to the protection or restoration of mangrove plants in those areas which are co-suitable for mangroves and S. alterniflora. In these areas, management measures should be conducted that hinder S. alterniflora invasions or clear existing S. alterniflora plants, firstly. This study provides guidance for the management of native species by preventing biological invasion.

BackgroundTriple-negative breast cancer (TNBC) is a subtype of breast cancer that is highly susceptible to recurrence and metastasis. The therapeutic drugs are limited due to the lack of effective biomarkers predicting the therapeutic efficacy and prognosis of disease. Previous studies have shown that platinum-containing regimens are effective for both early and advanced TNBC, therefore, it is particularly important to explore biomarkers for predicting the efficacy of platinum drugs and to screen the population sensitive to platinum drugs. Methods All patients with available tumor specimens from National Cancer Center in China were eligible for this prospective-retrospective study (n=189), including 149 patients with early TNBC (NCT01150513, CH-BC-007) and 40 patients with advanced TNBC (12-123/657, CH-BC-018) who are treated with or without platinum. The primary endpoint is disease-free survival (DFS) for early TNBC and progression free survival (PFS) for advanced TNBC. For HRD status and genomic signatures, indexed libraries were subjected to probe-based hybridization with a customized NGS panel targeting 733 cancer-related genes. 3DMed-HRD algorithm was evaluated based on loss of heterozygosity score (LOH), telomeric allelic imbalance score (TAI) and large-scale state transition score (LST) to characterize genomic instability using over 10,000 single-nucleotide polymorphisms distributed across human genome, adjusted by tumor ploidy and purity. HRD positive is defined by HRD score above the threshold (cut-off ≥30) and/or deleterious mutation in BRCA1/2. ResultsDeleterious BRCA1/2 mutations were detected in 21.2% (40/189) of TNBC patients and 48.1% (91/189) were defined as HRD positive. In advanced TNBC cohort, 21 patients received platinum-containing and 19 received platinum-free chemotherapy regimens for first-line chemotherapy. The progression-free survival (PFS) of the platinum-containing group was longer than that of the platinum-free group (median PFS 9.13 vs 2.97 months, HR 0.39, 95%CI, 0.19-0.81, P=0.011), and the PFS of patients with HRD positive was significantly longer than HRD negative patients (median PFS 13.6 vs 6.80 months, HR 0.38, 95%CI, 0.15-0.99, P=0.048) in platinum-containing group. Interestingly, HRD-positive patients have a significantly shorter PFS than HRD-negative in platinum-free group (median PFS 1.97 vs 4.52 months, HR 3.67, 95%CI, 1.20-11.22, P=0.023). For the HRD-positive patients, median PFS was significantly better in platinum-based group than platinum-free group (median PFS 13.6 vs 1.97 months, HR 0.12, 95%CI, 0.03-0.43, P=0.001). In early-stage TNBC cohort (n=149), 74 patients received platinum-containing and 75 received platinum-free chemotherapy regimens for adjuvant chemotherapy. In platinum-containing group, no statistically significant difference was found in DFS between HRD-positive and HRD-negative patients (P=0.118). High-risk TNBC group (Ki-67 ≥ 15%) analysis revealed that HRD-positive patients had a numerical better DFS than HRD-negative patients (HR 0.43, 95%CI, 0.12-1.50, P=0.180). Among patients with HRD-positive, patients in platinum-containing group had a tendency to benefit more than those in platinum-free group (HR 0.35, 95%CI, 0.11-1.10, P=0.062). Conclusions: We developed a novel algorithm to evaluate 3DMed-HRD status and highlights the potential utility of HRD in guiding chemotherapy for TNBC patients in this retrospective analysis. In comparison to platinum-free chemotherapy, the advanced TNBC HRD positive patients with advanced TNBC receiving platinum-based chemotherapy is a preferable regimen, and the HRD negative patients might be on the contrary. Prospective validation with larger sample size is needed. Citation Format: Yimeng Chen, Xue Wang, Feng Du, Jian Yue, Yiran Si, Lina Cui, Bei Zhang, Binghe Xu, Peng Yuan. The status of homologous recombination deficiency is a potential biomarker for platinum-based chemotherapy in triple-negative breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-08-12.

BACKGROUND: There is no clear clinical conclusion on whether assisted reproductive technology (ART) increases maternal and child risk and affects infant birth and development. This study aimed to perform a multiple regression analysis of the perinatal maternal and infant conditions in an assisted reproductive singleton pregnancy and the physical development and complications of such infants at 6 months old. METHODS: This study enrolled 145 singleton pregnant women who were admitted to Luohe Central Hospital between December 2017 and December 2019 to undergo in vitro fertilization and embryo transfer as a research group, and 160 singleton pregnant women who were naturally conceived at the same time and delivered at our hospital were selected as the control group. The relevant data of the patients were collected, and the perinatal conditions, neonatal complications, physical development and NBN score of infants aged 6 months were compared between the two groups. Multivariate logistic regression was used to analyze risk factors for perinatal complications in assisted reproduction singleton pregnancy. RESULTS: The incidence of complications in the perinatal period in the study group was significantly higher than that in the control group (P0.05). Likewise, at 6 months of age, the infants in the research group showed no significant difference to the control group in physical development indicators, including weight, head circumference, body mass index, height and NBN score (P>0.05). The analysis of the unconditional multivariate logistic regression model revealed hypertension during pregnancy, placental previa, premature membrane rupture, gestational age

Background and aimsCurrent treatment guidelines recommend ursodeoxycholic acid (UDCA) as the first-line treatment for new-diagnosed primary biliary cholangitis (PBC) patients. However, up to 40% patients are insensitive to UDCA monotherapy, and evaluation of UDCA response at 12 months may result in long period of ineffective treatment. We aimed to develop a new criterion to reliably identify non-response patients much earlier.MethodsFive hundred sixty-nine patients with an average of 59 months (Median: 53; IQR:32–79) follow-up periods were randomly divided into either the training (70%) or the validation cohort (30%). The efficiency of different combinations of total bilirubin (TBIL), alkaline phosphatase (ALP), and aspartate aminotransferase (AST) threshold values to predict outcomes was assessed at 1, 3 or 6 month after the initiation of UDCA therapy. The endpoints were defined as adverse outcomes, including liver-related death, liver transplantation and complications of cirrhosis. Adverse outcome-free survival was compared using various published criteria and a proposed new criterion.ResultsA new criterion of evaluating UDCA responses at 1 month was established as: ALP ≤ 2.5 × upper limit of normal (ULN) and AST ≤ 2 × ULN, and TBIL ≤ 1 × ULN (Xi’an criterion). The 5 year adverse outcome-free survival rate of UDCA responders, defined by Xi’an criterion, was 97%, which was significantly higher than that of those non-responders (64%). An accurate distinguishing high-risk patients’ capacity of Xi’an criterion was confirmed in both early and late-stage PBC.ConclusionsXi’an criterion has a similar or even higher ability to distinguish high-risk PBC patients than other published criteria. Xi’an criterion can facilitate early identification of patients requiring new therapeutic approaches.

Maize (Zea mays L.) is one of the most important crops in China. Since 2020, a new leaf spot disease has occurred in southwest China in areas such as Yunnan, Sichuan and Hubei provinces. Typical symptoms appeared after tasseling. The spots are scattered on the leaf surface, round to oval in shape with diameter range 3-20 mm. Spots are initially water-soaked, gradually turning yellow or white. In 2021, diseased leaf samples with typical white spot were collected for pathogen isolation and identification in Qujing, Yunnan province. Four small pieces of leaf tissue (about 0.25 cm2 in area) were excised from the edge of the necrotic lesion of each plant, surface sterilized with 75% ethanol for 1 min, rinsed three times with sterile distilled water, and soaked in sterile distilled water for 5 min. The solution was plated on Luria Broth medium (LB) plate (Shin et al. 2022) . After incubation at 28°C for 24 h, round, smooth-edged yellow colonies appeared in the LB plate. The bacterium isolated was gram-negative, negative for oxidase, positive for peroxidase, indole, citrate (Wells et al. 1987). Three strains (PA21QJ01, PA21QJ02 and PA21QJ03) showed identical colony morphology. PA21QJ01 was used for further molecular analyses. DNA was extracted from fresh colonies cultured in LB(Shin et al. 2022), and the fragments at the 16S rDNA, gyrB and rpoB loci were amplified using primers 27F/1492R (Galkiewicz and Kellogg 2008), UP-1/UP-2r (Yamamoto and Harayama 1995) and rpoBCM81-F/rpoBCM32b-R (Brady et al. 2008), respectively. The sequences of fragments of 16S rDNA, gyrB and rpoB from isolate PA21QJ01 were was deposited in GenBank (accession number: OM184301.1, OM302500, OM302499). A search for homologous sequences using BLAST resulted in 99.9, 99.6 and 99.8% identity of 16S rDNA, gyrB and rpoB, respectively, with sequences from the NN08200 of Pantoea ananatis (GenBank accession numbers: MK415050.1 for 16S rDNA, CP035034.1 for gyrB and CP035034.1.1 for rpoB). Above molecular results indicated that PA21QJ01 isolated from maize white spot is P. ananatis. Pathogenicity tests were performed on tasseled plants of the suscptible maize variety Wugu1790. After culture in LB medium plate at 30°C for 12 h, the bacterial solution was used for inoculation at a concentration of 1 × 108 CFU ml-1. After 7 days of inoculation, the leaves of the plants appeared water-soaked. After 10 days, white spot developed with brown margin. In contrast, the control plants remained healthy and symptomless. The same P. ananatis was reisolated in the inoculated maize plants, fulfilled Koch's law. In the last decade, P. ananatis has been reported to cause maize white spot in South Africa, Mexico, Poland, Argentina, Brazil (Sauer et al. 2015), and Ecuador (Toaza et al.2021). It has caused crop diseases with other crops, such as onion, rice, pineapple, melon, and sorghum, and others (Sauer et al. 2015). It caused leaf blight and leaf steak in rice in China (Yu et al. 2021). This is the first report of maize white spot caused by P. ananatis in China. However, to our knowledge, this is the first report of maize white spot disease in China. Attentions should be paid to screening for disease-resistant resources and breeding disease-resistant hybrids. Reference: Wells, J. M. et al. 1987. Int. J. Syst. Bacteriol. 37:136-143. Shin, G. Y. et al. 2022. Plant Dis. Doi: 10.1094/PDIS-08-21-1810-SC. Brady, C., et al. 2008. Syst. Appl. Microbiol. 31:447. Galkiewicz, J. P., and Kellogg, C. A. 2008. APPL ENVIRON MICROB, 74.24: 7828-7831. Toaza, A. et al. 2021. Plant Dis. Doi:10.1094/PDIS-02-21-0298-PDN Yamamoto, S., and Harayama, S. 1995. APPL ENVIRON MICROB, 61:1104.L. Sauer, A. V. et al. 2015. Agronomy Science and Biotechnology. Doi:10.33158/ASB.2015v1i1p21 Yu et al. 2021. Plant Dis. Doi:10.1094/PDIS-05-21-0988-PDN.

BACKGROUND This study sought to explore the effects of personalized rehabilitation exercises in the postoperative rehabilitation of breast cancer patients. METHODS A total of 93 breast cancer patients admitted to our hospital from January 2018 to December 2019 were taken as research subjects for this study. Based on order of admission, patients were assigned to a control group (46 cases) or an observation group (47 cases). All patients underwent a modified radical mastectomy for breast cancer. Patients in the control group received routine nursing and rehabilitation training after their operations, while those in the observation group received personalized rehabilitation exercises (for a period of 6 months) in addition to the rehabilitation training received by the control group. The immune function indicators, the incidence of upper limb-related complications, the movement of the shoulder joint, upper limb function, ability of daily living (ADL), and quality of life (QoL) was then compared between the two groups before and after the intervention. RESULTS Patients in the observation group had lower incidences of upper limb edema and subcutaneous fluid than those in the control group (P

BackgroundPrevious studies found homologous recombination deficiency (HRD) score could predict response to platinum-containing neoadjuvant chemotherapy in patients with triple negative breast cancer (TNBC), but its predictive effects of adjuvant chemotherapy is unexplored. Methods We developed the 3DMed-HRD algorithm, which combines loss of heterozygosity score (LOH), telomeric allelic imbalance score (TAI) and large-scale state transition score (LST) to characterize genomic instability using over 10000 SNPs, adjusted by tumor ploidy and purity. HRD positive is defined by deleterious mutation in BRCA1/2 or HRD score above the threshold (cut-off ≥30). Tumor samples were retrospectively obtained from 149 TNBC patients who received platinum-based (paclitaxel and cisplatin, TP) or platinum-free (cyclophosphamide and adriamycin, EC) adjuvant chemotherapy after surgery. All patients underwent the 3DMed-HRD testing. ResultsIn 25.5% (38/149) of TNBC patients deleterious germline or somatic mutation in BRCA 1/2 was detected. HRD results were available for all patients and 81of the 149 patients (54.4%) were defined as HRD positive. HRD-positive patients had younger age at diagnosis (median 46.7 years vs 51.5 years, P=0.002), higher differentiated (59.8% vs 39.5%; p=0.037), lower N-stage (55.7% vs 33.3%; p=0.006) and more common at high expression of Ki67 (64.7% vs 39.0%; p=0.015). In TP treatment group, patients with HRD positive tumors had a lower 5-year recurrence rate although the difference was not statistically significant (8.82% vs 16.1%; p=0.463). Patients in EC treatment cohort, 5-year recurrence rate was comparable between HRD positive and negative subgroups (15.4% vs 13.3%; p=1.0). In the 81 patients in HRD-positive cohort, patients in TP treatment group showed a numerical lower 5-year recurrence rates compared with patients in EC treatment group (8.82% vs 15.4%; p=0.489). Conclusions54.5% patients were characterized as HRD positive in TNBC patients in our cohort. HRD was associated with clinicopathologic characteristics. Our results suggested that HRD status mightguide chemotherapy treatment decisions. Prospective validation with larger sample size is needed. Citation Format: Xue Wang, Feng Du, Jian Yue, Yiran Si, Lina Cui, Peng Yuan. Homologous recombination deficiency score predicts patient characteristics and outcomes of triple negative breast cancer in China [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS4-46.

Cancers of all types have been shown to overexpress the carbohydrate-processing enzyme heparanase during angiogenesis and invasion, and this enzymatic activity correlates positively with tumor stage and the presence of metastases. While detection of heparanase enzymatic activity holds potential for cancer diagnosis and staging, no probe has been achieved for in vivo systems. We have recently developed fluorogenic heparanase probes for single-step detection of in vitro heparanase enzymatic activity, drastically simplifying the detection of heparanase activity. The modular chemistry allows us to extend the utility further to improve the sensitivity of in vitro heparanase activity detection and enable in vivo detection. Thus, we have developed a bioluminescent heparanase probe, which upon activation by heparanase produces a substrate of firefly luciferase, a standard reporter enzyme for in vivo monitoring of tumor progression. This probe will be validated in multi-well assays of heparanase activity, and applied in real-time correlation of heparanase activity and tumor progression in, fLuc+ tumors in animal models. These studies hold promise in enabling highly sensitive detection of heparanase activity both in vitro and in vivo, and in allowing future research of real-time monitoring of heparanase activity with the tumor progression. Citation Format: Johnathan Somers, Kelton Schleyer, Jun Liu, Zhishen Wang, Arjun Kafle, Lina Cui. A bioluminescent probe for detection and imaging of heparanase enzymatic activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2395.

Heparanase, an endo-β-glucuronidase, is the only known enzyme that is responsible for the cleavage of heparan sulfate chains of heparan sulfate proteoglycans. Its enzymatic activity plays a key role in remodeling and degradation of the extracellular matrix, generation of biologically active carbohydrate fragments, and liberation of effector molecules, such as growth factors and chemokines, that are bound to the heparan sulfate side chains. Overexpression of heparanase has been implicated in most types of cancers and various inflammatory diseases that usually involve extensive heparan sulfate degradation and extracellular matrix remodeling. In particular, the role of heparanase in cancer is highly significant as it regulates multiple biological activities that fosters tumor progression, angiogenesis, metastases. Consequently, heparanase has been considered a biological target for various therapeutic applications including anti-cancer drug development. However, one of the major challenges in this research area is the development of robust and feasible heparanase probes to study the enzymes’ fundamental biology in the extracellular matrix, to provide heparanase-targeted diagnostics, and to aid in drug discovery efforts. Our lab has been developing various small-molecule-based heparanase activatable probes, and we have successfully developed chemical strategies that allow sensitive detection of heparanase activity in various contexts. Two-photon microscopy allows for deeper tissue penetration, excellent spatial resolution, and efficient light detection with reduced photobleaching effects. Herein, we extend our chemical strategies to develop a heparanase-activatable two-photon fluorogenic probe for real-time detection of heparanase activity in animal models of highly-aggressive brain cancer using two-photon imaging. Citation Format: Zachary Rabinowitz, Zhishen Wang, Johnathan Somers, Jun Liu, Joyce Jiang, Nina Zhang, Lina Cui. Activatable fluorogenic probe for two-photon imaging of heparanase in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2394.

In this study, a finite element low-velocity impact model of Twaron®plain-woven fabric was created and analyzed using the commercial code ANSYS®-AUTODYN, and then was validated by drop-weight impact experiments. As a bullet or a fragment can strike a protective system from any angle in space, it is necessary to investigate fragment impact behavior response to impact threats from all angles of space. Therefore, in-plane obliquityθ, and spatial obliquityφ, were employed in this study and 17 different simulation test impact scenarios with different impact obliquity values were carried out using a standard hemispherical-head impactor. Results showed that the energy absorption of Twaron®fabric decreases with increasingθ, whereas under the sameθ, the energy absorption increases with increasingφ. This study also evaluated and compared the low-velocity impact performance of Twaron®fabric as a function of impactor shape, such as hemispherical, flat, and ogival heads, with differentθ. The results showed that under the same density, volume, and diameter conditions and at the normal impact scenario of a flat-head impactor, the fracture mechanism of the yarn is the same with all impact scenarios for a hemispherical-head impactor; the contacted yarns of the fabric fractured almost simultaneously. For the other oblique impact scenarios of the flat-head impactor, as well as impact scenarios of the ogival-head impactor, the yarns of the fabric fractured intermittently. Additionally, for the impact scenario with the ogival-head impactor, the effect of impact obliquity on energy absorption of the fabric was completely opposite to that of the hemispherical-head impact scenario. This is because in the hemispherical-head impact scenario, the fabric yarn tends to be damaged by tension, whereas in the ogival-head impact scenario, the fabric tends to be damaged by out-of-plane shear. These findings provide important guidance for the engineering of soft body armor and composite materials.

Heparanase (HPA) is a critical enzyme involved in the remodeling of the extracellular matrix (ECM), and its elevated expression has been linked with diseases such as various types of cancer and inflammation. The detection of heparanase enzymatic activity holds tremendous value in the study of the cellular microenvironment, and search of molecular therapeutics targeting heparanase, however, no structurally defined probes are available for the detection of heparanase activity. Here we present the development of the first ultrasensitive fluorogenic small-molecule probe for heparanase enzymatic activity via tuning the electronic effect of the substrate. The probe exhibits a 756-fold fluorescence turn-on response in the presence of human heparanase, allowing one-step detection of heparanase activity in real-time with a picomolar detection limit. The high sensitivity and robustness of the probe are exemplified in a high-throughput screening assay for heparanase inhibitors., Heparanase, a critical enzyme involved in the remodeling of the extracellular matrix, activates a disaccharide probe HADP to give a strong fluorescence signal.

In the present research, a physical-geometric-feature of continuous yarn in a plain woven fabric was created and its FE model was analysed by considering the two key issues of woven fabric, the crimp and inter-yarn friction. The basic parameters of Young’s modulus of single yarn and the inter-yarn friction coefficient were investigated for practical fabrics in tensile and pull-out tests. FE analysis indicated that the stress-strain curves of the FE model were effective in evaluating the equivalent modulus of a woven fabric by comparing with a tensile experiment on Twaron CT® Plain Woven Fabric. In addition, a simplified three dimensional model of the unit cell of plain woven fabric (UCPW) was employed to quantitively investigate two important fabric characteristics – the crimp rate of the yarn and inter-yarn friction-to determine their influence on the mechanical properties of the fabrics. Furthermore, we used FE analysis to evaluate how the crimp rate and inter-yarn friction affected the mechanical properties by determining the equivalent modulus of single yarn and UCPW in both uniaxial and biaxial tensile loading. The stresses at representative nodal points and the mechanical interaction between yarns were also investigated from a microscopic perspective, and their deformation mechanisms were also analysed and discussed.

This work proposes an approach to fabricate flexible transparent ultraviolet (UV)-shielding membrane by casting method, which uniformly disperses pristine zinc oxide nanoparticles (NPs) in low-density polyethylene (LDPE). The critical conditions for film fabrication, such as casting temperature, LDPE concentration in the solution, dissolution time, NP concentration, and post hot press cooling processes, are systematically studied. It is found that the casting temperature needs to be close to the melting temperature of LDPE, namely, 115°C, so that transparent film formation without cracks can be guaranteed. NP agglomerates are suppressed if the polymer concentration is controlled below 6%. For good dispersion of NPs, LDPE has to be swelled or unentangled enough in the solution (close to 200 h dissolution time), and then the NP agglomerates can be diminished due to the diffusion of the NPs into the polymer gel (322 h dissolution time). When the NPs are well-dispersed in the LDPE film, the film can completely shield UV light while allowing high transmissivity for the visible light. As the concentration of NPs in the film increases from 4 to 6%, the transmissivity of the film decreases, the tensile strength increases, and the tensile failure strain decreases.

Fluorophores experience altered emission lifetimes when incorporated into and liberated from macromolecules or molecular aggregates; this trend suggests the potential for a fluorescent, responsive probe capable of undergoing self-assembly and aggregation and consequently altering the lifetime of its fluorescent moiety to provide contrast between the active and inactive probes. We developed a cyanobenzothioazole-fluorescein conjugate (1), and spectroscopically examined the lifetime changes caused by its reduction-induced aggregation in vitro. A decrease in lifetime was observed for compound 1 in a buffered system activated by the biological reducing agent glutathione, thus suggesting a possible approach for designing responsive self-aggregating lifetime imaging probes.

Spermiogenesis is a complex process depending on the sophisticated coordination of a myriad of testis-enriched gene regulations. The regulatory pathways that coordinate this process are not well understood, and we demonstrate here that AXDND1, as a novel testis-enriched gene is essential for spermiogenesis and male fertility. AXDND1 is exclusively expressed in the round and elongating spermatids in humans and mice. We identified two potentially deleterious mutations of AXDND1 unique to non‐obstructive azoospermia (NOA) patients through selected exonic sequencing. Importantly, Axdnd1 knockout males are sterile with reduced testis size caused by increased germ cell apoptosis and sloughing, exhibiting phenotypes consistent with oligoasthenoteratozoospermia. Axdnd1 mutated late spermatids showed head deformation, outer doublet microtubules deficiency in the axoneme, and loss of corresponding accessory structures, including outer dense fiber (ODF) and mitochondria sheath. These phenotypes were probably due to the perturbed behavior of the manchette, a dynamic structure where AXDND1 was localized. Our findings establish AXDND1 as a novel testis-enrich gene essential for spermiogenesis and male fertility probably by regulating the manchette dynamics, spermatid head shaping, sperm flagellum assembly.

Substrate-based probes utilize known substrate specificity parameters to create a probe that can be activated by a target enzyme. In developing probes for heparanase, an endo-ß-glucuronidase, we previously reported that small, inactive substrate-based probes could be electronically tuned by incorporating electron-withdrawing atoms on the aromatic aglycone fluorophore, ortho- to the cleaved glycosidic bond. However, the installation of electron-withdrawing groups directly onto established fluorophores or other reporters complicates the synthesis of new heparanase probes. In this work we report a new design strategy to expand the toolkit of heparanase imaging probes, in which the installation of an electronically tuned benzyl alcohol linker restored the activity of a previously inactive heparanase probe using 4-methylumbelliferone as the fluorescent reporter, suggesting such a linker can provide a scaffold for facile development of activatable heparanase probes bearing a variety of imaging moieties.

Hydrolases are important molecules that are involved in a wide range of biological functions and their activities are tightly regulated in healthy or diseased states. Detecting or imaging the activities of hydrolases, therefore, can reveal underlying molecular mechanisms in the context of cells to organisms, and their correlation with different physiological conditions can therefore be used in diagnosis. Due to the nature of hydrolases, substrate-based probes can be activated in their catalytic cycles, and cleavage of covalent bonds frees reporter moieties. For test-tube type bulk detection, spatial resolution is not a measure of importance, but for cell- or organism-based detection or imaging, spatial resolution is a key factor for probe sensitivity that influences signal-to-background ratio. One strategy to improve spatial resolution of the probes is to form a covalent linkage between the reporter moiety and intracellular proteins upon probe activation by the enzyme. In this work, we developed a generalizable linker chemistry that would allow in situ labeling of various imaging moieties via quinone methide species. To do so, we synthesized probes containing a monofluoromethyl or a difluoromethyl groups for β-galactosidase activation, while using fluorescein as a fluorescent reporter. The labeling efficacy of these two probes was evaluated in vitro. The probe bearing a monofluormethyl group exhibited superior labeling efficiency in imaging β-galactosidase activity in living cells. This study provides a versatile linker for applying quinone methide chemistry in the development of hydrolase-targeting probes involving in situ labeling.

Male ‘blind sterile’ mice with the causative TBC1 domain family member 20 (TBC1D20) deficiency are infertile with excessive germ cell apoptosis and spermatogenesis arrest at the spermatid stage. Sertoli cells are characterised as ‘nurse cells’ essential for normal spermatogenesis, but the role and corresponding molecular mechanisms of TBC1D20 deficiency in Sertoli cells of mice are not clear to date. In the present study, the histopathology of the testis and Sertoli cell proliferation and apoptosis were determined, and the corresponding molecular mechanisms were investigated by western blotting. Our data showed that TBC1D20 exhibits a testis-abundant expression pattern, and its expression level is positively associated with spermatogenesis. TBC1D20 is assembled in the Golgi and endoplasmic reticulum and is widely expressed by various germ cell subtypes and Sertoli cells. TBC1D20 deficiency in Sertoli cells led to an excessive apoptosis ratio and G1/S arrest. The increased apoptosis of TBC1D20-deficient Sertoli cells resulted from caspase-12 activation. TBC1D20-deficient Sertoli cells had an abnormal Golgi-endoplasmic reticulum structure, which led to endoplasmic reticulum stress, resulting in cell cycle arrest and excessive apoptosis. It suggested that TBC1D20 deficiency triggers irreversible endoplasmic reticulum stress resulting in G1/S arrest and excessive apoptosis in TBC1D20-deficient Sertoli cells, and TBC1D20 deficiency in Sertoli cells may also contribute to the infertility phenotype in ‘blind sterile’ male mice.

Heparanase is an enzyme highly involved in the progression of many types of cancer through its enzymatic processing of heparan sulfate proteoglycans. Unfortunately, research and clinical investigation of this target is hampered by the lack of simple, effective tools to detect its enzymatic activity. Despite the natural preference of heparanase to cleave polysaccharide substrates, we previously developed the first disaccharide-based fluorogenic probe for heparanase enzymatic activity, overcoming this deficiency in existing tools. However, the chemistry of the probe synthesis required installation of electron-withdrawing fluorine atoms on the fluorophore to facilitate activation by heparanase, a design strategy that excludes the use of many common imaging reporters. To accommodate a wider variety of imaging modalities for heparanase detection, we experimented with the addition of a collapsible linker with suitable electronegativity that can be easily tuned. Our results show that a tetra-fluorinated benzyl alcohol linker enables heparanase activation of a probe bearing any reporter, giving comparable results to our original fluorinated coumarin probe. This disaccharide-linker scaffold holds potential for accessing a broader scope of heparanase imaging probes for laboratory and clinical applications of this burgeoning biomarker. Citation Format: Kelton Schleyer, Jun Liu, Zhishen Wang, Hua Guo, Lina Cui. Expanding the library of small molecule imaging probes for heparanase [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2925.

Cellular senescence is a mechanism by which a cell enters stable cell cycle arrest triggered by various types of stress such as telomere shortening, oxidative stress and DNA damage. It is implicated in a number of pathophysiological conditions including age-related diseases, degenerative disorders and cancer. Despite its importance, the current toolbox for studying or monitoring senescence in vivo is limited. We have developed a series of novel near-infrared (NIR) fluorescent probes that can be activated by senescence-associated β-galactosidase (SA-β-Gal) and self-immobilize onto intracellular proteins via in situ labeling. We compared the labeling efficiency of these probes in vitro and in cells and the results shows that by modulating the leaving group, we can improve the labeling efficiency of the probes and enhance their retention at the site of activation. These probes can be used for imaging of senescence developed in live cells, as well as long-term tracking of senescent cells in various animal models. Citation Format: Zixin Chen, Yuzhao Zhang, Seyedehalaleh Anvar, Chang-hung Wu, Jun Liu, Joyce Z. Jiang, Lina Cui. NIR fluorescent probes for senescence imaging via tunable in situ labeling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 138.

Background: Ovarian cancer (OC) is a highly lethal gynecologic malignancy, neoadjuvant chemotherapy (NACT) with interval de-bulking surgery (IDS) is an established strategy for woman with advanced epithelial ovarian cancer (EOC). Tumor infiltrating lymphocytes (TILs) play an important role in the pathogenesis of OC. However, the effect of NACT on tumor immune microenvironment (TIME) has not been evaluated. Methods: All patients underwent NACT with IDS. Formalin-fixed paraffin-embedded tissue samples were collected from biopsy specimens and the corresponding post-NACT surgical specimens. The densities and percentage of specific TILs [CD8+, CD3+, CD4+, CD20+, tumor associated macrophages (TAMs), dim CD56+NK, bright CD56+NK, PD-1+ etc.] within the TIME were analyzed by multiplex immunohistochemistry (3DMedcines Inc.). Serum CA 125 was measured 3-weekly per protocol and evaluated the response to NACT using GCIG CA125 response criteria. Results: A total of 28 patients were enrolled in this study with planned IDS after NACT. Of 25 patients evaluated by GCIG CA125 response criteria, 76% (19/25) had a CA125 response. In CA125 response patients, we observed significant recruitments of the densities and percentage of CD8+ in tumor, percentage of PD-1+ cell in tumor after-NACT vs before-NACT (34.85/mm3 vs 110.54/mm3, p=0.0258; 0.50% vs 1.73%, p=0.0132; 0.53% vs 2.66%, p=0.0576, respectively), while no significant change was found in TAMs, NK cells and CD20+ cells. However, in CA125 non-response patients, the TILs in tumor did not change significantly. Conclusion: NACT could release the tumor associated antigen by killing tumor cells for uptake by tumor-infiltrating dendritic cells and promote the infiltration of CD8+ and PD-1+ T lymphocyte in CA125 response OC patients. Given the limited clinical prognostic information and the number of patients in this study, the results of this study will need confirmation in larger studies in the future. Citation Format: Guangming Cao, Dingchao Hua, Lina Cui, Xiaochen Zhao, Shuzhen Wang, Lei Zhu. The effect of neoadjuvant chemotherapy on tumor immune microenvironment in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6106.

Background: Chemotherapy is the most common treatment strategy for triple negative breast cancer (TNBC) patients. Nevertheless, due to adverse drug reactionsthe molecular high heterogeneity of TNBC and no appropriate meaningful efficacy markers, it is still difficult to establish preferred therapeutic strategies and predict the outcomes for TNBC. This study was to investigate the potential predictors and therapeutic targets based on genetic features. Methods: A total of 386 TNBC patients were randomized 1:1 to receive either six cycles of paclitaxel + cisplatin (TP) or four cycles of epirubicin + cyclophosphamide followed by four cycles of docetaxel (EC-T) adjuvant chemotherapy after surgery (NCT01150513), which were described previously. Finally, 149 TNBC patients with clinical and tumor sequencing data availability were retrospectively analysed by NGS for 733 cancer-related genes. All tumor samples were collected during the surgical operation and subjected to NGS for evaluating genomic mutations and the potential predictors. Cox regression model and Kaplan-Meier method were applied to evaluate disease-free survival (DFS). Results: In the surgical cohort receiving adjuvant chemotherapy, 74 patients received platinum and 75 received platinum-free chemotherapy as adjuvant chemotherapy. The most frequently mutated genes in this surgical TNBC cohort were TP53 (84%), BRCA1 (18%), BRCA2 (15%), POL1 (13%), PTEN (12%), REV3L (11%), FANCC (10%), and PARP4 (10%). We analyzed the associations between 733 cancer-related gene mutations and DFS after adjuvant chemotherapy. For the TP group, PIK3CA mutation (19%, 14/74) was discovered to correlate with poor DFS for patients treated with platinum-containing adjuvant therapy (HR=3.2, P=0.037), and KMT2D mutation (15%, 11/75) display worse DFS for patients treated with EC-T platinum-free group (HR=3.0, p=0.039). However, BRCA1/2 mutation (24%, 35/149) was found to be associated with poor prognosis (HR=2.1 (95% CI: 1-4.6), p=0.047), irrespective of therapeutic regimen. Conclusions: In this population of surgical TNBC patients, NGS analysis identified potential predictive markers. PIK3CA mutation in TP platinum-containing group and KMT2D mutation in EC-T platinum-free group were significantly associated to poor prognosis for adjuvant chemotherapy in triple negative breast cancer. Citation Format: Xue Wang, Feng Du, Jian Yue, Yiran Si, Lina Cui, Bei Zhang, Xiaochen Zhao, Binghe Xu, Peng Yuan. KMT2D and PIK3CA mutation as potential factors to predict adjuvant chemotherapy efficacy in surgical triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5084.

Recent studies have evidenced that ubiquitin-specific proteases (USPs) are associated with the occurrence and drug resistance of T-cell acute lymphoblastic leukemia (T-ALL). N6-methyladenosine (m6A) demethylase AlkB homolog 5 (ALKBH5) exerts a carcinogenic effect in human cancers and improves the mRNA stability of USPs. Whether ubiquitin specific protease 1 (USP1) controls chemoresistance in T-ALL is unknown. Our study demonstrated that USP1, Aurora kinase B (Aurora B) and ALKBH5 levels were highly expressed in glucocorticoid (GC)-resistant T-ALL patients and cells (CEM-C1). High expression of USP1 was correlated to the poor prognosis of T-ALL patients. Silencing USP1 increased CEM-C1 cell sensitivity to dexamethasone (Dex), reduced cell invasion, promoted cell apoptosis, and ameliorated glucocorticoid receptor (GR) expression. USP1 mediated T-ALL chemoresistance by interacting with Aurora B. Overexpression of USP1 reversed the promotive effects of Aurora B inhibitor on the sensitivity of CEM-C1 cells to Dex, cell apoptosis and GR level and the inhibition effect on cell invasion. Downregulation of ALKBH5 reduced the levels of USP1 and Aurora B, facilitated CEM-C1 cell sensitivity to Dex, apoptosis and GR expression, suppressed cell invasion. However, overexpression of USP1 reversed all the effects of ALKBH5 on CEM-C1 cells. In vivo results showed that tail vein injection of sh-USP1 resulted in a significant prolongation of mouse survival and maintained the normal weight of mice compared to the Dex group, reduced USP1 expression and facilitated GR expression. In conclusion, downregulation of USP1 ameliorated glucocorticoid resistance of T-cell acute lymphoblastic leukemia cells through suppressing Aurora B expression and elevating GR level.