A novel stoploss mutation CYB5R3 c.906A>G(p.*302Trpext*42) involved in the pathogenesis of hereditary methemoglobinemia.

Recessive congenital methemoglobinemia (RCM) is a hereditary autosomal disorder with an extremely low incidence rate. Here, we report a case of methemoglobinemia type I in a patient with congenital persistent cyanosis. The condition was attributed to a novel compound heterozygous mutation in CYB5R3, characterized by elevated methemoglobin levels (13.4 % of total hemoglobin) and undetectable NADH cytochrome b5 reductase (CYB5R3) activity. Whole-exome sequencing (WES) revealed two heterozygous mutations in CYB5R3: a previously reported pathogenic missense mutation c.611G>A(p.Cys204Tyr) inherited from the father, and a novel stop codon mutation c.906A>G(p.*302Trpext*42) from the mother, the latter mutation assessed as likely pathogenic according to ACMG guidelines. In cells overexpressing the CYB5R3 c.906A>G mutant construct, the CYB5R3 mRNA level was significantly lower than in cells overexpressing the wild-type (WT) CYB5R3 construct. However, there was no significant difference in protein expression levels between the mutant and WT constructs. Notably, an additional protein band of approximately 55 kDa was detected in the mutant cells. Immunofluorescence localization showed that, compared to wild-type CYB5R3, the subcellular localization of the CYB5R3 p.*302Trpext*42 mutant protein did not show significant changes and remained distributed in the endoplasmic reticulum and mitochondria. However, the c.906A>G(p.*302Trpext*42) mutation resulted in increased intracellular reactive oxygen species (ROS) levels and decreased NAD+/NADH ratio, suggesting impaired CYB5R3 function and implicating this novel mutation as likely pathogenic.

A Liquid Band-Aid with Mesenchymal Stem Cell-Derived Exosomes for Wound Healing in Mice.

INTRODUCTION/OBJECTIVE: This study aimed to examine the effect of a human umbilical cord mesenchymal stem cell-derived exosome (hUC-MSC-Exo) liquid band-aid on wound healing in mice. METHODS: hUC-MSC-Exos were prepared from the supernatant via ion exchange chromatography. The composition ratio of the chitosan liquid band-aid was optimized to form a film and encapsulate hUC-MSC-Exo. The biological effects of chitosan exosome liquid band-aid on human umbilical vein endothelial cells (HUVECs) were observed, and its anti-bacterial properties were tested. BALB/c mice with back skin injury were randomly divided into chitosan exosome liquid band-aid group (CS-Exo), chitosan liquid band-aid group (CS), and normal saline control group (Con), and wound healing was evaluated post-treatment. Skin tissue samples posttreatment were collected for H&E staining. RESULTS: The hUC-MSC-Exo was prepared and characterized. The optimum conditions for film formation were 1% chitosan solution and 15% poloxamer 407/poloxamer 188 (pH 5.0 ~ 7.0). The chitosan exosome liquid band-aid promoted HUVEC proliferation and migration and markedly inhibited Escherichia coli and Staphylococcus aureus growth in vitro. In vivo, the wound healing rate in the CS-Exo group was higher than that in the Con and CS groups. Fourteen days post-treatment, the wounds completely healed, and hair grew normally, which was consistent with H&E results. Mouse weights in each group did not change significantly after administration, indicating that the chitosan exosome liquid band-aid had no obvious toxic side effects. CONCLUSION: Local chitosan exosome liquid band-aid application can promote wound healing in mice, and the mechanism could be related to hUC-MSC-Exo-induced vascular endothelial cell proliferation and migration.

RBC-hitchhiking PLGA nanoparticles loading ß-glucan as a delivery system to enhance in vitro and in vivo immune responses in mice.

Red blood cells (RBCs) naturally trap some bacterial pathogens in the circulation and kill them by oxidative stress. Following neutralization, the bacteria are presented to antigen-presenting cells in the spleen by the RBCs. This ability of RBCs has been harnessed to develop a system where they play a crucial role in enhancing the immune response, offering a novel approach to enhance the body's immunity. In this work, a conjugate, G-OVA, was formed by connecting ß-glucan and OVA through a disulfide bond. Poly (lactic-co-glycolic acid) (PLGA) was then employed to encapsulate G-OVA, yielding G-OVA-PLGA. Finally, the nanoparticles were adsorbed onto RBCs to develop G-OVA-PLGA@RBC. The results demonstrated that the delivery of nanoparticles by RBCs enhanced the antibody response to antigens both in vitro and in vivo. The objective of this study was to investigate the increased immune activity of G-OVA-PLGA nanoparticles facilitated by RBCs transportation and to elucidate some of its underlying mechanisms. These findings are anticipated to contribute valuable insights for the development of efficient and safe immune enhancers.

Identification of potential biomarkers of cuproptosis in cerebral ischemia.

Objective: Cerebral ischemia can cause mild damage to local brain nerves due to hypoxia and even lead to irreversible damage due to neuronal cell death. However, the underlying pathogenesis of this phenomenon remains unclear. This study utilized bioinformatics to explore the role of cuproptosis in cerebral ischemic disease and its associated biomarkers. Method: R software identified the overlap of cerebral ischemia and cuproptosis genes, analyzed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), and explored hub genes. Expressions and localizations of hub genes in brain tissue, cells, and immune cells were analyzed, along with predictions of protein structures, miRNAs, and transcription factors. A network was constructed depicting hub gene co-expression with miRNAs and interactions with transcription factors. Ferredoxin 1 (FDX1) expression was determined using qRT-PCR. Results: Ten cuproptosis-related genes in cerebral ischemia were identified, with GO analysis revealing involvement in acetyl-CoA synthesis, metabolism, mitochondrial function, and iron-sulfur cluster binding. KEGG highlighted processes like the tricarboxylic acid cycle, pyruvate metabolism, and glycolysis/gluconeogenesis. Using the Human Protein Atlas, eight hub genes associated with cuproptosis were verified in brain tissues, hippocampus, and AF22 cells. Lipoyl(octanoyl) transferase 1 (LIPT1), was undetected, while others were found in mitochondria or both nucleus and mitochondria. These genes were differentially expressed in immune cells. FDX1, lipoic acid synthetase (LIAS), dihydrolipoamide S-acetyltransferase (DLAT), pyruvate dehydrogenase E1 component subunit alpha 1 (PDHA1), PDHB, and glutaminase (GLS) were predicted to target 111 miRNAs. PDHA1, FDX1, LIPT1, PDHB, LIAS, DLAT, GLS, and dihydrolipoamide dehydrogenase (DLD) were predicted to interact with 11, 10, 10, 9, 8, 7, 5, and 4 transcription factors, respectively. Finally, FDX1 expression was significantly upregulated in the hippocampus of ovariectomized rats with ischemia. Conclusion: This study revealed an association between cerebral ischemic disease and cuproptosis, identifying eight potential target genes. These findings offer new insights into potential biomarkers for the diagnosis, treatment, and prognosis of cerebral ischemia, and provide avenues for the exploration of new medical intervention targets.

Corrigendum: Regional disparities in the prevalence and correlated factors of myopia in children and adolescents in Gansu, China.

[This corrects the article DOI: 10.3389/fmed.2024.1375080.].

Advances in the self-organized total synthesis of natural products.

Natural product total synthesis has trailblazed in the era of multistep synthesis. The strategic application of existing synthetic methodologies and the stepwise construction that revolves around newly developed, tailored key steps, are two basic tactics in the principle of classic retrosynthetic analysis. However, a new synthetic model, termed self-organized total synthesis, has emerged in recent years, enabling the rapid creation of specific natural products by a one-pot reaction. Distinct from conventional analysis associated with certain bond disconnections, the design of self-organized total synthesis focuses on seeking a series of self-organized reaction sequences which can be integrated compatibly under a uniform condition, therefore allowing the entire sequence to proceed in one pot, and most importantly, starting from commercially available feedstocks or biomass materials. Whilst dauntingly challenging, this synthetic strategy is more consistent with the biogenetic pathway of natural products compared with conventional counterparts, and will hopefully provide the shortest synthesis for such natural products. Through this rational analysis, one-pot total synthesis is no longer in the way of serendipity but can be precisely designed and manipulated. In this review, we account for the definition, delimitation, and categorization of self-organized total synthesis and then elucidate a comprehensive understanding of this synthetic strategy based on our intensive explorations. We also highlight the contributions of other research groups in this growing field and anticipate that it will give rise to advancing new methodologies, as well as new concepts within organic synthesis.

Cell and transcriptomic diversity of infrapatellar fat pad during knee osteoarthritis.

OBJECTIVES: In this study, we employ a multiomic approach to identify major cell types and subsets, and their transcriptomic profiles within the infrapatellar fat pad (IFP), and to determine differences in the IFP based on knee osteoarthritis (KOA), sex and obesity status. METHODS: Single-nucleus RNA sequencing of 82 924 nuclei from 21 IFPs (n=6 healthy control and n=15 KOA donors), spatial transcriptomics and bioinformatic analyses were used to identify contributions of the IFP to KOA. We mapped cell subclusters from other white adipose tissues using publicly available literature. The diversity of fibroblasts within the IFP was investigated by bioinformatic analyses, comparing by KOA, sex and obesity status. Metabolomics was used to further explore differences in fibroblasts by obesity status. RESULTS: We identified multiple subclusters of fibroblasts, macrophages, adipocytes and endothelial cells with unique transcriptomic profiles. Using spatial transcriptomics, we resolved distributions of cell types and their transcriptomic profiles and computationally identified putative cell-cell communication networks. Furthermore, we identified transcriptomic differences in fibroblasts from KOA versus healthy control donor IFPs, female versus male KOA-IFPs and obese versus normal body mass index (BMI) KOA-IFPs. Finally, using metabolomics, we defined differences in metabolite levels in supernatants of naïve, profibrotic stimuli-treated and proinflammatory stimuli-treated fibroblasts from obese compared to normal BMI KOA-IFPs. CONCLUSIONS: Overall, by employing a multiomic approach, this study provides the first comprehensive map of the cellular and transcriptomic diversity of human IFP and identifies IFP fibroblasts as key cells contributing to transcriptomic and metabolic differences related to KOA disease, sex or obesity.

Risk Factors of Cytomegalovirus Retinitis Occurrence After Allogeneic Hematopoietic Stem Cell Transplantation.

PURPOSE: To explore the potential risk factors for the occurrence of human cytomegalovirus (HCMV) retinitis (CMVR) in allogeneic hematopoietic stem cell transplantation (allo-HSCT) patients. METHODS: This is a retrospective, nested case-control study conducted in hematological patients with CMVR who underwent allo-HSCT. Patients diagnosed with CMVR after allo-HSCT were included as the case group, and those without CMVR were matched by a ratio of 1:2 and were recruited as controls. We selected 19 pre- and post-transplant indicators for univariate analysis between the cases and controls, and then Logistic regression analysis was used to calculate the odds ratio (OR) and 95% confidence intervals (CI) for exploration of risk factors of the CMVR occurrence. RESULTS: A total of 1308 allo-HSCT patients from January 1, 2020 to July 31, 2023 were analyzed, and 27 patients were diagnosed CMVR with a median onset time of 222 days after transplantation. In univariate analysis, donors of stem cells source, HLA-match types (including matched sibling donor, haploidentical donor, and unrelated donor), post-transplant Epstein-Barr virus (EBV) viremia, platelet implantation time, and serostatus of CMV-IgG were more easily to develop CMVR than controls (p < 0.001, p = 0.003, p < 0.001, p = 0.032, p = 0.038, respectively). Multivariate logistic regression analysis showed that stem cells source (OR 7.823, 95% CI 1.759-34.800), HLA-match types (OR 7.452, 95% CI 1.099-50.542), and post-transplant EBV infection (OR 7.510, 95% CI 1.903-29.640) were positively associated with the onset of CMVR. CONCLUSION: Stem cells derived from bone marrow and peripheral blood, HLA-match types, and post-transplant EBV viremia are important risk predictors of CMVR in allo-HSCT patients. These results suggest that clinicians should pay more attention to these indicators when formulating preventive measures pre- and post-transplant.

A Surface-Reconstructed Bilayer Heterojunction Enables Efficient and Stable Inverted Perovskite Solar Cells.

The efficiency of perovskite photovoltaics remains distant from their theoretical limits, primarily due to high photovoltage losses. Here a strategy is reported to minimize voltage losses by reconstructing the perovskite surface into a bilayer heterojunction (BLH) structure. Unlike conventional low-dimensional capping layers, typically constrained to a few nanometers to prevent low fill factors, this methodology facilitates a more comprehensive reaction with surface defects, allowing a more substantial capping layer (≈50 nanometers) without compromising charge transport integrity. Time-resolved microwave conductivity analysis indicates a significant reduction in trap density at the top region of the perovskite film, showing an order of magnitude lower than that of the pristine sample. Incorporating this BLH in inverted cells results in a remarkably low photovoltage deficit of 325 mV, leading to a power conversion efficiency (PCE) of 26.1% (25.72% certified). The encapsulated device maintains 94% of its original efficiency after 1200 h of maximum power point tracking under one sun illumination at 65 °C.

Quantitative detection of circular RNA and microRNA at point-of-care using droplet digital CRISPR/Cas13a platform.

Circular RNA (circRNA) and microRNA (miRNA) are both non-coding RNAs (ncRNAs) that serve as biomarkers for cancer diagnosis and prognosis. Quantitative detection of these ncRNAs is of particular importance to elucidate the functional mechanisms and evaluate their potential as biomarkers. However, the inherent structures of circRNA and miRNA are different from the mRNA, conventional qRT-PCR is unsuitable for the detection of these ncRNAs. Here, we propose a sensitive method for quantitative detection of circRNA and miRNA using polydisperse droplet digital CRISPR/Cas13a (PddCas13a). It can achieve limits of detection (LOD) as low as ∼10 aM without polymerase-based amplification. To efficiently detect the circRNA and miRNA in real samples, we use a chemically modified crRNA to enhance the stability of crRNA and improve the performance of Cas13a in complex environments containing contaminants. By integrating an extraction-free procedure with PddCas13a, we experimentally demonstrate the applicability of PddCas13a by testing clinical samples. Furthermore, we develop an automated and portable instrument for PddCas13a and verify its applicability for the detection of circRNA and miRNA from exosomes in point-of-care (POC) setting. This is the first report to detect the circRNA and miRNA simultaneously in POC setting. We envision this platform could promote the research of ncRNAs.

Human activities reshape the spatial overlap between North Chinese leopard and its wild ungulate prey.

BACKGROUND: Rapidly expanding human activities have profoundly changed the habitat use of both large carnivores and their prey, but whether and how human activities affect the interactions between them has received relatively less attention. In this study, we conducted a systematically designed camera-trapping survey on an endangered large carnivore (North Chinese leopard Panthera pardus japonensis) and its wild ungulate prey (Siberian roe deer Capreolus pygargus and wild boar Sus scrofa) in the Taihang Mountains of central North China. Using conditional two-species occupancy model based on data derived from the extensive sampling effort (15,654 camera-days at 102 camera sites), we examined the relationship of spatial use between leopards and each prey species under the effects of human presence, free-ranging cattle, roads and settlements. RESULTS: Humans and cattle had contrasting effects on the relationship of spatial use between leopard and roe deer, with higher and lower spatial segregation between them at human and cattle-frequented sites, respectively. Roads might create a shelter for wild boar from leopard predation, with less spatial segregation between them at sites close to the roads. CONCLUSIONS: Our findings demonstrate that human activities are reshaping the spatial overlap between large carnivores and their prey, and have non-equivalent effects among different types of human activity. Such effects may further alter the strength of interspecific interactions between predator and prey, with far-reaching influences on the community and ecosystem that require more research.

Protocol for high-resolution 3D visualization of insect regenerating legs through micro-computed tomography.

Appendage regeneration occurs within the opaque exoskeleton in arthropods, making it challenging to visualize the regenerative processes dynamically. In this protocol, we present a strategy to scan and capture the high-resolution details of microstructural tissues at certain regeneration points through micro-computed tomography (micro-CT). We describe steps for tissue preparation, fixation, critical point drying, micro-CT scanning, and 3D visualization. This approach promises significant utility in the field of regeneration, particularly in studies involving arthropods such as insects and crustaceans. For complete details on the use and execution of this protocol, please refer to Ren et al.1.

A review of deep learning methods for gastrointestinal diseases classification applied in computer-aided diagnosis system.

Recent advancements in deep learning have significantly improved the intelligent classification of gastrointestinal (GI) diseases, particularly in aiding clinical diagnosis. This paper seeks to review a computer-aided diagnosis (CAD) system for GI diseases, aligning with the actual clinical diagnostic process. It offers a comprehensive survey of deep learning (DL) techniques tailored for classifying GI diseases, addressing challenges inherent in complex scenes, clinical constraints, and technical obstacles encountered in GI imaging. Firstly, the esophagus, stomach, small intestine, and large intestine were located to determine the organs where the lesions were located. Secondly, location detection and classification of a single disease are performed on the premise that the organ's location corresponding to the image is known. Finally, comprehensive classification for multiple diseases is carried out. The results of single and multi-classification are compared to achieve more accurate classification outcomes, and a more effective computer-aided diagnosis system for gastrointestinal diseases was further constructed.

Screening for Helicobacter pylori to Prevent Gastric Cancer: A Pragmatic Randomized Clinical Trial.

Importance: Effects of screening for Helicobacter pylori on gastric cancer incidence and mortality are unknown. Objective: To evaluate the effects of an invitation to screen for H pylori on gastric cancer incidence and mortality. Design, Setting, and Participants: A pragmatic randomized clinical trial of residents aged 50 to 69 years in Changhua County, Taiwan, eligible for biennial fecal immunochemical tests (FIT) for colon cancer screening. Participants were randomized to either an invitation for H pylori stool antigen (HPSA) + FIT assessment or FIT alone. The study was conducted between January 1, 2014, and September 27, 2018. Final follow-up occurred December 31, 2020. Intervention: Invitation for testing for H pylori stool antigen. Main Outcomes and Measures: The primary outcomes were gastric cancer incidence and gastric cancer mortality. All invited individuals were analyzed according to the groups to which they were randomized. Results: Of 240 000 randomized adults (mean age, 58.1 years [SD, 5.6]; 46.8% female), 63 508 were invited for HPSA + FIT, and 88 995 were invited for FIT alone. Of the 240 000 randomized, 38 792 who were unreachable and 48 705 who did not receive an invitation were excluded. Of those invited, screening participation rates were 49.6% (31 497/63 508) for HPSA + FIT and 35.7% (31 777/88 995) for FIT alone. Among 12 142 participants (38.5%) with positive HPSA results, 8664 (71.4%) received antibiotic treatment, and eradication occurred in 91.9%. Gastric cancer incidence rates were 0.032% in the HPSA + FIT group and 0.037% in the FIT-alone group (mean difference, -0.005% [95% CI, -0.013% to 0.003%]; P = .23). Gastric cancer mortality rates were 0.015% in the HPSA + FIT group and 0.013% in the FIT-alone group (mean difference, 0.002% [95% CI, -0.004% to 0.007%]; P = .57). After adjusting for differences in screening participation, length of follow-up, and patient characteristics in post hoc analyses, an invitation for HPSA + FIT was associated with lower rates of gastric cancer (0.79 [95% CI, 0.63-0.98]) but not with gastric cancer mortality (1.02 [95% CI, 0.73-1.40]), compared with FIT alone. Among participants who received antibiotics, the most common adverse effects were abdominal pain or diarrhea (2.1%) and dyspepsia or poor appetite (0.8%). Conclusions and Relevance: Among residents of Taiwan, an invitation to test for HPSA combined with FIT did not reduce rates of gastric cancer or gastric cancer mortality, compared with an invitation for FIT alone. However, when differences in screening participation and length of follow-up were accounted for, gastric cancer incidence, but not gastric cancer mortality, was lower in the HSPA + FIT group, compared with FIT alone. Trial Registration: ClinicalTrials.gov Identifier: NCT01741363.

Effect of intravenous different drugs on the prevention of restlessness during recovery period of pediatric laparoscopic surgery: a randomized control trial.

PURPOSE: To explored the impact of dexmedetomidine and esketamine in mitigating restlessness during the postoperative recovery phase following laparoscopic surgery in children. METHODS: 102 individuals aged 1 to 7 years experiencing laparoscopic surgery were randomly allocated into three groups, each accepting 1 µg/kg of dexmedetomidine, 0.3 mg/kg of esketamine, or saline immediately at the end of carbon dioxide pneumoperitoneum. Emergence agitation (EA) occurrence was assessed by PAED scale and 5-point agitation scale. Pain was judged using Face, Legs, Activity, Cry, and Consolability (FLACC) scale. The recovery time, extubation time, and post-anesthesia care unit (PACU) stay time were recorded for all three groups. RESULTS: Patients administered 1 µg/kg of dexmedetomidine (8.8%) and individuals given 0.3 mg/kg of esketamine (11.8%) showed lower incidences of emergence agitation compared to those receiving saline (35.5%; P = 0.009). There was no statistically significant difference in the time to discharge from the PACU among the three groups of patients (P > 0.05). The recovery time and extubation time were notably extended in the dexmedetomidine group (40.88 ± 12.95 min, 42.50 ± 13.38 min) when compared to the saline group (32.56 ± 13.05 min, 33.29 ± 11.30 min; P = 0.009, P = 0.010). CONCLUSION: Following CO2 pneumoperitoneum in pediatric laparoscopic surgeries, the intravenous administration of 1 µg/kg dexmedetomidine or 0.3 mg/kg esketamine effectively lowers EA occurrence without extending PACU time.

An investigation into the cognitive flexibility of undergraduate dental hygiene students at two dental schools in Asia.

Background/purpose: Cognitive flexibility is a personality trait, which can influence how effectively a healthcare professional can manage a challenging clinical situation. This study explored the cognitive flexibility of undergraduate dental hygiene students at two universities in Asia to gather baseline information in order to consider whether there was an educational need for pre-clinical students with regards to this personality factor. Materials and methods: Participants were all undergraduate dental hygiene students in the 2022 academic year at Tokyo Medical and Dental University (TMDU; n = 79) and Taipei Medical University (TMU; n = 88). An anonymous questionnaire was distributed, which included questions on demographic information and the Cognitive Flexibility Inventory (CFI). Results: The response rates were 97.5% (TMDU) and 89.8% (TMU). The CFI total scores were 91.77 for TMDU and 95.87 for TMU, and there was no significant difference between the two schools. The Alternatives scores were significantly higher in TMU than TMDU (P = 0.044), but there were no significant differences in the Control scores between the two schools. There were also no significant differences between school years of both universities. Conclusion: The dental hygiene students from both dental schools exhibit moderate cognitive flexibility. The educators in both dental schools may consider whether they should investigate and develop this personality factor further during the undergraduate training of their dental hygiene students.

Association of probiotics, prebiotics, synbiotics or yogurt supplement with prevalence and all-cause mortality of depression: NHANES 2005-2016.

BACKGROUND: A growing body of studies revealed that enteric dysbacteriosis could result in depression via the "gut-microbiota-brain axis" (GMBA). Whether probiotics, prebiotics, and synbiotics supplements could lessen the risk of depression is a topic attracting attention. This research was conducted to evaluate the relationship between probiotics, prebiotics, synbiotics, or yogurt supplements and depression with large cross-sectional data. METHODS: All data in our research was sourced from the National Health and Nutrition Examination Survey (NHANES) (2005-2016). Probiotics, prebiotics, synbiotics, and yogurt supplements were identified using Food Frequency Questionnaire (FFQ) and Dietary Supplement Use 30-Day (DSQ). We employed the Patient Health Questionnaire (PHQ-9) for evaluating depression. Logistic regression and the Kaplan-Meier curve were performed to examine the correlation between the supplements and depression, as well as mortality. RESULTS: A total of 17,745 adult participants were selected. The participants who supplemented probiotics, prebiotics, synbiotics, or yogurt products in the last 30 days showed a significantly lower depression rate compared with those who didn't. Specifically, the supplements could alleviate depressive symptoms including sad, anhedonia, sleep problems, fatigue, appetite changes, and psychomotor changes. This association was more prominent in specific populations such as the population aged 40-60 years, male, whites. The supplements also show more significant effects on increasing survival rates in patients with mild depression. LIMITATION: Cross-sectional analysis reveals correlative but not causative association. CONCLUSION: Based on the analysis of NHANES data, our research highlights the positive effect the supplements have on preventing depression, relieving depressive symptoms and increasing survival rates. This effect varied across populations.

Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid Tetrastichus hagenowii (Ratzeburg).

Parasitoid wasps play a crucial role in the efficient control of pests, a substantial menace to human health and well-being. Tetrastichus hagenowii (Ratzeburg) stands out as the most effective egg parasitoid wasp for controlling American cockroaches, but accurate and stable reference genes for quantitative real-time polymerase chain reaction of T. hagenowii genes are still lacking. In this study, we assessed seven candidate nuclear genes, including α-tubulin (α-TUB), elongation factor-1-alpha (EF-1α), ß-actin (Actin), ribosomal protein 49 (RP49), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), nicotinamide adenine dinucleotide (NADH), and elongation factor 2 (EF2) of T. hagenowii. By analyzing expression stability with four algorithms (Delta Ct, geNorm, NormFinder, and BestKeeper), as well as comprehensive ranking with RefFinder, we identified α-TUB as the most stable reference gene for the larval, pupal, female adult, and male adult stages. Subsequently, we estimated the transcript levels of vitellogenin (Vg) and cuticle protein (CP) after normalization with α-TUB across various developmental stages. Significantly higher expression levels of CP and Vg were observed in pupae and female adults, respectively, consistent with previous findings in other insects. This study offers a reliable reference gene for normalizing transcription levels of T. hagenowii genes.