The high mutation rate of the influenza virus poses a significant challenge to global health, highlighting the urgent need for broad-spectrum vaccines. Here, we engineered Ad-Hex, a pan-influenza vaccine comprising three chimpanzee adenoviral vectors (Ad-H1H3, Ad-BYBV, and Ad-H5H7), each encoding the hemagglutinin (HA) genes from two distinct influenza virus subtypes/lineages in the △E1 region. A single intranasal dose of Ad-Hex induced robust humoral, cellular, and mucosal immunity, conferring complete protection against lethal challenge with six vaccine-matched strains in female C57BL/6 or Balb/c mice. Notably, the vaccine achieved 60

The tumor-stroma ratio (TSR) has emerged as a promising prognostic biomarker in epithelial ovarian cancer (EOC); however, its preoperative assessment remains challenging. To develop a non-invasive CT-based radiomics machine learning model for preoperative TSR prediction and to evaluate its association with platinum resistance and survival outcomes in EOC. This retrospective study included 172 patients with pathologically confirmed EOC. TSR was histologically classified as stroma-rich (≥ 50

Background:Modulating the cGAS-STING pathway by bioactive nanodevices is a promising strategy for combating infection-associated inflammatory disorders. However, the development of pharmacological inhibitors for cGAS-STING signaling is currently hindered by lacking cell-specific targeting capability. This study aimed to develop a potent, drug-free nanodevice that specifically targets pulmonary macrophages to modulate the cGAS-STING pathway for ameliorating infection-associated detrimental lung inflammation. Methods:Cigarette smoke extract-modified peptide gold nanoparticle hybrids (CSE-P12) were synthesized. Transcriptomic analysis, western blotting, autophagy reporter assays, and confocal microscopy were employed to assess the effects of CSE-P12 on gene expression, STING degradation, autophagic flux, and inflammation. TEM imaging and LC-MS/MS were utilized to elucidate the molecular mechanisms underlying CSE-P12-induced autophagy in macrophages. Finally, the HAdV4-induced pneumonia and CLP-induced sepsis models on wild-type and STING-/- mice were used to evaluate the therapeutic efficacy of CSE-P12 and validate its inhibitory mechanisms on the cGAS-STING pathway. Results:CSE-P12 nanodevices are extensively internalized by macrophages via energy-dependent cellular uptake. This large internalization triggers autophagic degradation of STING, thereby effectively inhibiting the cGAS-STING-mediated interferon responses and inflammation. In the HAdV4-induced viral pneumonia mouse model, intratracheally instilled CSE-P12 effectively targets pulmonary macrophages, suppresses STING activation, and significantly alleviates lung inflammation and injury. The depletion of the pulmonary macrophages abolishes these protective effects. The therapeutic potential of CSE-P12 is further validated in a CLP-induced polymicrobial sepsis mouse model, where it significantly prolongs mouse survival and decreases lung inflammation. Conclusions:CSE-P12 effectively targets pulmonary macrophages and exhibits potent anti-inflammatory activities in viral pneumonia and sepsis-induced acute lung injury by inducing autophagic flux to facilitate STING degradation. This work provides a new paradigm for designing targeted nanotherapeutics to modulate STING activation in inflammatory diseases.

2'-fucosyllactose (2'-FL) holds significant role in the infants' nutrition. While microbial production of 2'-FL has predominantly utilized Escherichia coli and Saccharomyces cerevisiae, the potential of Pichia pastoris, renowned for its robust NADPH regeneration capability, remains underexplored. Herein, we systematically engineered the metabolism of P. pastoris to develop an efficient 2'-FL-producing cell factory. We first constructed the de novo biosynthesis pathway for 2'-FL in P. pastoris, achieving an initial titer of 0.143 g/L. By optimizing enzyme selection and solubility of α-1,2-fucosyltransferase (FutC), 2'-FL production was enhanced by nearly ten folds. Subsequently, engineering NADPH supply further increased the 2'-FL production by 170 %. Furthermore, we enhanced energy supply by incorporating an orthogonal energy module based on the methanol dissimilation pathway and increasing GTP availability, resulting in a 32 % improvement in 2'-FL production. Finally, through the optimization of fermentation condition, we realized the production titer of 2'-FL to 3.50 g/L in shake-flask, representing the highest titer in P. pastoris. These findings highlight the potential of P. pastoris as a chassis to produce chemicals by providing abundant NADPH and utilizing methanol as co-substrate to supply sufficient energy.

Identification and verification of clinically actionable molecular variations to refine currently adopted risk-stratified treatment strategy for esophageal squamous cell carcinoma (ESCC) is urgently needed. Here, we evaluated FGFR3 amplification status by fluorescence in situ hybridization (FISH) performed on tissue microarrays and its prognostic value in 526 ESCC patients. FGFR3 amplification was found in 3.0

Highly effective and broad-spectrum influenza vaccines are urgently required to prevent influenza outbreaks. Hemagglutinin (HA), M2 ectodomain (M2e), and nucleoprotein (NP) are crucial target antigens for the development of universal influenza vaccines. To generate a novel multivalent influenza vaccine, the HA genes of influenza B Yamagata (BY) and Victoria (BV) strains, and the NP gene of H1N1 were cloned into the E1 region of the chimpanzee adenoviral vector, AdC68, and M2e epitopes of H1N1 and H3N2 were fused to the loop region of the AdC68 fiber, resulting in the recombinant adenoviral vector vaccine, AdC-Flu-Tet. The immunoprotective effects of AdC-Flu-Tet were evaluated in the mouse models. The results showed that AdC-Flu-Tet successfully induced robust humoral and cellular immune responses and conferred full protection against H1N1, H3N2, BY, and BV infections. In conclusion, AdC-Flu-Tet is a promising candidate as a novel influenza vaccine with high protective efficacy.

DExH-Box helicase 9 (DHX9), also known as RNA helicase A (RHA), belongs to the DExD/H-Box superfamily II of helicases. Due to its fundamental roles in processes such as transcription and maintenance of genomic stability, DHX9 has emerged as a key regulator in various cancer types. Specifically, microsatellite instable (MSI) cancers, such as MSI-high (MSI-H) colorectal cancer, and tumors with defective homologous recombination exhibit a strong dependency on DHX9, suggesting that targeting DHX9 could offer a significant therapeutic opportunity. Employing fragment-based drug design, we have discovered GH3595, a novel allosteric DHX9 inhibitor that selectively targets the DHX9 protein. In vitro, GH3595 displayed a low nM potency in ATPase and helicase activity assays, which translated to specific low nM inhibition of MSI-H colorectal cancer and BRCA loss of function breast cancer proliferation. In vivo, GH3595 demonstrated favorable pharmacokinetic properties in different species and showed an acceptable safety profile. Furthermore, daily oral dosing of GH3595 led to the regression of MSI-H xenograft tumors without adverse effects on body weight. More importantly, GH3595 induced dose-dependent intra-tumoral circular RNA induction, showing a well-correlated PK/PD/Efficacy relationship. In summary, we report the discovery and characterization of GH3595, a potent and selective inhibitor of DHX9, which effectively disrupts DHX9-mediated signaling pathways. These findings highlight its significant therapeutic potential for the treatment of malignancies characterized by microsatellite instability or deficiencies in homologous recombination. Faridoon Faridoon, Jiapeng Li, Jiyue Zheng, Xiang Wang, Guiping Zhang. Discovery of GH3595: a novel allosteric DHX9 inhibitor for cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4247.

Current engineered synthetic scaffolds fail to functionally repair and regenerate ruptured native tendon tissues, partly because they cannot satisfy both the unique biological and biomechanical properties of these tissues. Ideal scaffolds for tendon repair and regeneration need to provide porous topographic structures and biological cues necessary for the efficient infiltration and tenogenic differentiation of embedded stem cells. To obtain crimped and porous scaffolds, highly aligned poly(l-lactide) fibers were prepared by electrospinning followed by postprocessing. Through a mild and controlled hydrogen gas foaming technique, we successfully transformed the crimped fibrous mats into three-dimensional porous scaffolds without sacrificing the crimped microstructure. Porcine derived decellularized tendon matrix was then grafted onto this porous scaffold through fiber surface modification and carbodiimide chemistry. These biofunctionalized, crimped, and porous scaffolds supported the proliferation, migration, and tenogenic induction of tendon derived stem/progenitor cells, while enabling adhesion to native tendons. Together, our data suggest that these biofunctionalized scaffolds can be exploited as promising engineered scaffolds for the treatment of acute tendon rupture.

Despite prolonged surveillance and interventions, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza viruses continue to pose a severe global health burden. Thus, we developed a chimpanzee adenovirus-based combination vaccine, AdC68-HATRBD, with dual specificity against SARS-CoV-2 and influenza virus. When used as a standalone vaccine, intranasal immunization with AdC68-HATRBD induced comprehensive and potent immune responses consisting of immunoglobin (Ig) G, mucosal IgA, neutralizing antibodies, and memory T cells, which protected the mice from BA.5.2 and pandemic H1N1 infections. When used as a heterologous booster, AdC68-HATRBD markedly improved the protective immune response of the licensed SARS-CoV-2 or influenza vaccine. Therefore, whether administered intranasally as a standalone or booster vaccine, this combination vaccine is a valuable strategy to enhance the overall vaccine efficacy by inducing robust systemic and mucosal immune responses, thereby conferring dual lines of immunological defenses for these two viruses.

The increase in population-related and environmental issues has emphasized the need for more efficient and sustainable production strategies for foods and chemicals. Carbohydrates are macronutrients sourced from crops and undergone transformation into various products ranging from foods to chemicals. Continuous efforts have led to the identification of a promising hybrid system that couples the electrochemical reduction of CO 2 to intermediates containing one to three carbons (C 1–3 ) with the transformation of the intermediates using engineered microorganisms into valuable products. Here we use yeast to transform C 1–3 substrates into glucose and structurally tailored glucose derivatives, such as the sugar alcohol myo-inositol, the amino monosaccharide glucosamine, the disaccharide sucrose and the polysaccharide starch. By metabolic rewiring and mitigation of glucose repression, the titre of glucose and sucrose reached dozens of grams per litre. These results provide directions for microbial sugar-derived foods and chemicals production from renewable reduced CO 2 -based feedstocks.

Classic chimeric hemagglutinin (cHA) was designed to induce immune responses against the conserved stalk domain of HA. However, it is unclear whether combining more than one HA head domain onto one stalk domain is immunogenic and further induce immune responses against influenza viruses. Here, we constructed numerous novel cHAs comprising two or three fuzed head domains from different subtypes grafted onto one stalk domain, designated as cH1-H3, cH1-H7, cH1-H3-H7, and cH1-H7-H3. The three-dimensional structures of these novel cHAs were modelled using bioinformatics simulations. Structural analysis showed that the intact neutralizing epitopes were exposed in cH1-H7 and were predicted to be immunogenic. The immunogenicity of the cHAs constructs was evaluated in mice using a chimpanzee adenoviral vector (AdC68) vaccine platform. The results demonstrated that cH1-H7 expressed by AdC68 (AdC68-cH1-H7) induced the production of high levels of binding antibodies, neutralizing antibodies, and hemagglutinin inhibition antibodies against homologous pandemic H1N1, drifted seasonal H1N1, and H7N9 virus. Moreover, vaccinated mice were fully protected from a lethal challenge with the aforementioned influenza viruses. Hence, cH1-H7 cHAs with potent immunogenicity might be a potential novel vaccine to provide protection against different subtypes of influenza virus.

Abstract Objective：The aim of this study is to share our experience in the conservative treatment of duodenal perforation in children. Method：Retrospective collection of clinical data on 6 children with duodenal perforation treated at Fujian Children's Hospital from May 2022 to October 2023. Six cases were all males, ranging in age from 8 to 14 years old, and the onset time ranged from 4 to 24 hours. Main manifestations are abdominal pain, vomiting, fever, etc. Among them, 5 children have undergone abdominal CT examination at local hospitals, indicating gastrointestinal perforation. Result：All 6 patients were diagnosed with perforation of the anterior wall of the duodenum, of which 5 cases were confirmed by ultrasound examination to be located in the bulb, and 1 case was located at the junction of the bulb and the descending part. The perforation diameter of all cases was less than 1cm.Four of them were less than 5mm, and 1 case was 6.9mm. Abdominal X-ray plain film showed 3 cases of free gas downstream of the diaphragm. Five cases underwent conservative treatment, including fasting, gastrointestinal decompression, antibiotics, proton pump inhibitor (PPI), octreotide, and nutritional support. Blood routine monitoring showed that white blood cell (WBC) count and C-reactive protein (CRP) level gradually returned to normal. They recovered after 10-16 days of hospitalization. One case underwent emergency laparoscopic repair of duodenal perforation due to sepsis after 2 days of conservative treatment. Complications such as chyloperitoneum, pleural effusion and subphrenic effusion occurred after the surgery, and the patient recovered after 34 days of hospitalization. All patients recovered and were discharged. Continue to take omeprazole acid therapy for 8 weeks after discharge. All cases were followed up for a period of 1 month to 1 year. Only 1 patient underwent gastroscopy 3 months later and, diagnosed with multiple duodenal ulcers (S2 stage), and continued treatment. The remaining 5 cases showed no signs of second time perforation, bloody stools, abdominal pain, or vomiting. Conclusion：Duodenal perforation in children is rare and often caused by primary ulcers. Most of them has a history of chronic abdominal pain, with symptoms of abdominal pain, vomiting, fever, and signs of peritonitis during acute attacks. Abdominal X-ray, abdominal CT plain scan, and ultrasound can assist in diagnosis. As long as there is no progression to sepsis, conservative treatment can achieve the same therapeutic effect as surgical treatment, with satisfactory outcomes and acceptable hospital stay.

Biomaterials have evolved from inert materials to responsive entities, playing a crucial role in disease diagnosis, treatment, and modeling. However, their advancement is hindered by limitations in chemical and mechanical approaches. Synthetic biology enabling the genetically reprograming of biological systems offers a new paradigm. It has achieved remarkable progresses in cell reprogramming, engineering designer cells for diverse applications. Synthetic biology also encompasses cell-free systems and rational design of biological molecules. This review focuses on the application of synthetic biology in theranostics, which boost rapid development of advanced biomaterials. We introduce key fundamental concepts of synthetic biology and highlight frontier applications thereof, aiming to explore the intersection of synthetic biology and biomaterials. This integration holds tremendous promise for advancing biomaterial engineering with programable complex functions.

Background:Rabies is a lethal zoonotic disease that kills approximately 60,000 people each year. Although inactivated rabies vaccines are available, multiple-dose regimensare recommended for pre-exposure prophylaxis or post-exposure prophylaxis,which cuts down the cost- and time-effectiveness, especially in low- and middle incomecountries.Methods:We developed a nucleoside-modified Rabies mRNA-lipid nanoparticle vaccine (RABV-G mRNA-LNP) encoding codon-optimized viral glycoprotein and assessed the immunogenicity and protective efficacy of this vaccine in mice comparing to a commercially available inactivated vaccine.Results:We first showed that, when evaluated in mice, a single vaccination of RABV-G mRNA with a moderate or high dose induces more potent humoral and T-cell immune responses than that elicited by three inoculations of the inactivated vaccine. Importantly, mice receiving a single immunization of RABV-G mRNA, even at low doses, showed full protection against the lethal rabies challenge. We further demonstrated that the humoral immune response induced by single RABV-G mRNA vaccination in mice could last for at least 25 weeks, while a two-dose strategy could extend the duration of the highly protective response to one year or even longer. In contrast, the three-dose regimen of inactivated vaccine failed to do so.Conclusion:Our study confirmed that it is worth developing a single-dose nucleoside-modified Rabies mRNA-LNP vaccine, which could confer much prolonged and more effective protection.

Inflammatory myofibroblastic tumor (IMT) is a mesenchymal neoplasm of intermediate biologic potential, which occurs mostly in the lung and abdomen cavity of children and young adults. Uterine IMTs are rare. Herein, we presented clinicopathologic features of 4 uterine IMTs. All four patients were initially diagnosed as leiomyosarcoma by other hospitals and corrected to uterine IMT after pathological consultation. Patient age ranged from 44 to 64 years old. Two cases demonstrated multiple masses. Microscopically, three tumors were composed of fascicular spindled cells with eosinophilic cytoplasm, and the other one was densely composed of spindled and epithelioid cells with bizarre and multinucleated cells. Tumor cells showed variable nuclear atypia, ranging from mild to severe. Prominent inflammatory cell infiltration was found in one case, and necrosis in two tumors. Immunochemistry staining revealed expression of smooth muscle markers in all four tumors, including a-SMA and desmin. Three tumors were positive for ALK protein expression. FISH analysis demonstrated ROS1 rearrangement in one tumor and ALK rearrangement in the other 3 tumors. NGS analysis showed novel NUDCD3-ROS1 and NRP2-ALK fusions in two tumors and TNS1-ALK fusion in the other two tumors. Gene aberrations involving p53 signaling pathway were identified in all four cases. All patients received surgery as primary treatment, and one had neoadjuvant chemotherapy. Three patients recurred within 12 months, and the other one recurred after 7 years. Patients with recurrence were treated with a combination of chemotherapy, targeted therapy, or surgery. In conclusion, the diagnosis of uterine IMTs can be challenging. Ancillary studies including ALK IHC, FISH, and NGS are helpful to establish diagnosis and to discover novel gene rearrangement potentially for targeted therapy.

Background/purposeRhubarb peony decoction (RPD) is a formula of traditional Chinese medicine that has been widely used to treat intra-abdominal inflammatory diseases. To investigate the therapeutic efficacy of RPD in pediatric periappendiceal abscess, patients who received intravenous antibiotics alone were compared with those treated with intravenous antibiotics combined with RPD.MethodsA retrospective review of children with periappendiceal abscess who received conservative treatment in our hospital between January 2013 and April 2022 was performed. The patients were divided into an intravenous antibiotic group (the control group) and an intravenous antibiotic combined with RPD group (the intervention group). Interval appendectomy (IA) was generally performed 10–12 weeks after conservative treatment. The primary outcome was the cure rate of conservative treatment, while the secondary outcomes included the recurrence rate, days of total intravenous antibiotic use, length of hospital stay (LOS), postoperative complications, and liver injury caused by RPD.ResultsA total of 142 patients (77 girls and 65 boys) were included, 52 in the control group and 90 in the intervention group. The two groups were similar in demographic data and clinical characteristics (P > 0.05). The mean total course of RPD in the intervention group was 11.82 days. The intervention group had a significantly higher cure rate than the control group (93.33% vs. 80.77%, P = 0.029), and the length of total intravenous antibiotic use (P = 0.150), LOS (P = 0.077), recurrence rate (9.52% vs. 4.76%, P = 0.439), as well as the operation time (P = 0.101), LOS (P = 0.572), and postoperative complications (P = 0.549) were not significantly different between the two groups when the patients received IA. No patient had a liver injury caused by RPD during the treatment.ConclusionIntravenous antibiotics combined with RPD demonstrated high effectiveness and safety for treating pediatric periappendiceal abscess.

Cell proliferation requires the integration of catabolic processes to provide energy, redox power and biosynthetic precursors. Here we show how the combination of rational design, metabolic rewiring and recombinant expression enables the establishment of a decarboxylation cycle in the yeast cytoplasm. This metabolic cycle can support growth by supplying energy and increased provision of NADPH or NADH in the cytosol, which can support the production of highly reduced chemicals such as glycerol, succinate and free fatty acids. With this approach, free fatty acid yield reached 40% of theoretical yield, which is the highest yield reported for Saccharomyces cerevisiae to our knowledge. This study reports the implementation of a synthetic decarboxylation cycle in the yeast cytosol, and its application in achieving high yields of valuable chemicals in cell factories. Our study also shows that, despite extensive regulation of catabolism in yeast, it is possible to rewire the energy metabolism, illustrating the power of biodesign.

The methylotrophic budding yeast Pichia pastoris has been utilized to the production of a variety of heterologous recombinant proteins owing to the strong inducible alcohol oxidase promoter (pAOX1). However, it is difficult to use P. pastoris as the chassis cell factory for high-valuable metabolite biosynthesis due to the low homologous recombination (HR) efficiency and the limitation of handy selective markers, especially in the condition of multistep biosynthetic pathways. Hence, we developed a novel CRISPR/Cas9 system with highly editing efficiencies and recyclable auxotrophic selective marker (HiEE-ReSM) to facilitate cell factory in P. pastoris. Firstly, we improved the HR rates of P. pastoris through knocking out the non-homologous-end-joining gene (Δku70) and overexpressing HR-related proteins (RAD52 and RAD59), resulting in higher positive rate compared to the basal strain, achieved 97%. Then, we used the uracil biosynthetic genes PpURA3 as the reverse screening marker, which can improve the recycling efficiency of marker. Meanwhile, the HR rate is still 100% in uracil auxotrophic yeast. Specially, we improved the growth rate of uracil auxotrophic yeast strains by overexpressing the uracil transporter (scFUR4) to increase the uptake of exogenous uracil from medium. Meanwhile, we explored the optimal concentration of uracil (90 mg/L) for strain growth. In the end, the HiEE-ReSM system has been applied for the inositol production (250 mg/L) derived from methanol in P. pastoris. The systems will contribute to P. pastoris as an attractive cell factory for the complex compound biosynthesis through multistep metabolic pathway engineering and will be a useful tool to improve one carbon (C1) bio-utilization.

Residual tumors after surgery lead to a high risk of recurrence, and effective strategies clearing the residual tumors urgently need to be developed. Locally administering antitumor agents after surgery has been proved effective to clear residual tumor cells, but achieving long-lasting efficacy and repeatable treatment are still challenges for these agents. In this study, we orderly arranged propyl gallate (PG)-grafting MXene onto calcium-alginate hydrogel (CA) layer to develop a photothermal-responsive biopaster for the tumor post-surgery recurrence prevention. The order arrangement of PG-grafting MXene nanosheets with the prolonged degradation time (compared to the non-PG-grafting ones) could retain the photothermal responsiveness for 14 days. Therefore, this biopaster provided long-term photothermal therapeutic capability to kill the residual tumors and effectively prevented the tumor recurrence (after surgery). This work provides references for designing NIR-responsive 2D nanomaterials with enhanced NIR-responsiveness and long-term antitumor efficacy.