Genkwanin alleviates intervertebral disc degeneration via regulating ITGA2/PI3K/AKT pathway and inhibiting apoptosis and senescence.

Intervertebral disc degeneration (IVDD) is a progressive degenerative disease influenced by various factors. Genkwanin, a known anti-inflammatory flavonoid, has not been explored for its potential in IVDD management. This study aims to investigate the effects and mechanisms of genkwanin on IVDD. In vitro, cell experiments revealed that genkwanin dose-dependently inhibited Interleukin-1ß-induced expression levels of inflammatory factors (Interleukin-6, inducible nitric oxide synthase, cyclooxygenase-2) and degradation metabolic protein (matrix metalloproteinase-13). Concurrently, genkwanin upregulated the expression of synthetic metabolism genes (type II collagen, aggrecan). Moreover, genkwanin effectively reduced the phosphorylation of phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin, mitogen-activated protein kinase (MAPK), and nuclear factor-κB (NF-κB) pathways. Transcriptome sequencing analysis identified integrin α2 (ITGA2) as a potential target of genkwanin, and silencing ITGA2 reversed the activation of PI3K/AKT pathway induced by Interleukin-1ß. Furthermore, genkwanin alleviated Interleukin-1ß-induced senescence and apoptosis in nucleus pulposus cells. In vivo animal experiments demonstrated that genkwanin mitigated the progression of IVDD in the rat model through imaging and histological examinations. In conclusion, This study suggest that genkwanin inhibits inflammation in nucleus pulposus cells, promotes extracellular matrix remodeling, suppresses cellular senescence and apoptosis, through the ITGA2/PI3K/AKT, NF-κB and MAPK signaling pathways. These findings indicate that genkwanin may be a promising therapeutic candidate for IVDD.

Validity of the modified versions of SARC-F+EBM for sarcopenia screening and diagnosis in China: the PPLSS study.

BACKGROUND AND OBJECTIVES: It is recommended by Asian Working Group for Sarcopenia to early identify people at risk for sarcopenia using simple screening tools like SARC-F. The modified version SARC-F+EBM showed higher diagnostic performance. However, this cut-off value of body mass index (BMI) remained uncertain to be used in Chinese population. In this study, we used appropriate BMI recommended for Chinese older population and further modified SARC-F+EBM by combining calf circumference. METHODS AND STUDY DESIGN: Diagnostic tests were performed and the receiver operating characteristics analyses were conducted between the SARC-F, SARC-F+EBM (cut-off of BMI: ≤ 21 kg/m2), SARC-F+EBM (CN) (cut-off of BMI: ≤ 22 kg/m2), SARC-CalF and SARC-CalF+EBM (CN) (cut-off of BMI: ≤ 22 kg/m2) in 1660 community-dwelling participants aged ≥ 65 years from China. RESULTS: The participants had an average age of 71.7±5.1 years, of which 56.8% were women. All the modified models could enhance the areas under the receiver operating characteristic curve (AUC) of original SARC-F (all p<0.001). The SARC-F+EBM (CN) also showed a significantly higher sensitivity of 47.4% (p<0.001) and an AUC of 0.809 (p=0.005) than SARC-F+EBM. SARC-CalF+EBM (CN) was validated to be of great diagnostic value of the highest AUC of 0.88 among these sarcopenia screening tools, including SARC-F, SARC-CalF and SARC-F+EBM (CN) (all p<0.001). Using this study population as a reference, the optimal cut-off value of SARC-CalF+EBM (CN) is ≥12 points, with a sensitivity of 79.3% and a specificity of 80.7%. CONCLUSIONS: The SARC-F+EBM (CN) and SARC-CalF+EBM (CN) could enhance the diagnostic performance of SARC-F and SARC-F+EBM and are suitable sarcopenia screening tools for Chinese population.

A randomized controlled study investigating the efficacy of electro-acupuncture and exercise-based swallowing rehabilitation for post-stroke dysphagia: Impacts on brainstem auditory evoked potentials and cerebral blood flow.

BACKGROUND: Swallowing rehabilitation behavioral therapy and traditional Chinese acupuncture therapy are widely used in the treatment of post-stroke dysphagia (PSD). This study investigated the therapeutic effect of electro-acupuncture combined with exercise-based swallowing rehabilitation on PSD and its effect on brainstem auditory evoked potential (BAEP) and cerebral blood flow. METHODS: The 120 PSD patients were divided into 2 groups (n = 60 each) by simple random grouping method, that is, an experimental and control group, receiving routine swallowing training, or additional intervention with electro-acupuncture at a frequency of 5 times/week. Data in swallowing function, BAEP, and cerebrovascular color Doppler ultrasound parameters were collected before treatment, as well as after treatment. An intergroup comparison was conducted using an independent sample t-test, and an intra-group comparison was conducted among different time points using a paired t-test. The data were analyzed using the SPSS Statistics 22.0 software; P < .05 was considered statistically significant. RESULTS: The therapeutic effects were significantly better in the experimental group compared with the control group (P < .05). The standard swallowing function assessment scores were significantly lower in both groups after treatment (P < .05), and the score in the observation group was lower than in the control group (P < .05). The peak latency of BAEP waves III and IV, and the inter-peak latency between peaks III to V and I to V in the 2 groups changed significantly (P < .05). The peak systolic velocity (PSV), end-diastolic velocity (EDV), and mean velocity (MV) were significantly increased in both groups after treatment (P < .05). The pulsatility index decreased significantly in both groups (P < .05), and the PSV, EDV, and MV were higher in the experimental group than in the control group (P < .05). CONCLUSION: Electro-acupuncture, combined with swallowing training in the treatment of Post-stroke Dysphagia, effectively improved cerebral microcirculation and conduction velocity, enhanced the motor function of swallowing muscles, and promoted the recovery of swallowing function.

Hydrogeochemical Disparities and Constraints of Water Produced from Various Coal Seams in the Baode Block, Ordos Basin, China.

The hydrogeochemical characteristics of coalbed water play a crucial role in assessing the production level of coalbed methane (CBM) due to its involvement in the entire process of CBM generation, migration, accumulation, and extraction. To investigate variations in hydrochemical characteristics and controlling factors among different coal seams, a representative CBM field (Baode block) within the Ordos basin in China was chosen as a target. We have systematically collected produced water samples from coal seams of the Permian Shanxi Formation (P1s) and Taiyuan Formation (P1t). Tests and analyses were conducted on conventional cation and anions, trace elements, pH value, total dissolved solids (TDS), stable isotopes of hydrogen and oxygen in water, and inorganic carbon (δD, δ18O, and δ13CDIC). The findings indicate that the P1s coal seam primarily contains HCO3-Na type water, while the P1t coal seam consists of Cl-Na and HCO3-Na types of water. The disparity in water types between P1s and P1t can be attributed to interactions between water and rocks. The isotopic compositions of δD, δ18O, and δ13CDIC suggest that the sampled coalbed waters originate from atmospheric precipitation, with subsequent microbial activity. It is suggested that TDS content along with bicarbonate concentration can serve as effective indicators for determining high productivity due to weaker hydraulic conditions and a more enclosed water environment in P1t coal seams; threshold values being >1000 mg/L for TDS and >10 mequiv/L for bicarbonate concentration. Additionally, microbial activity is found to be more widespread in P1t compared to P1s. Principal component analysis reveals a significantly higher contribution of conventional ions toward TDS content observed within the P1t coal seam compared to that of P1s coal seam, accompanied by alterations in pH control parameters. The water produced from the P1s coalbed is primarily controlled by evaporite and silicate weathering/dissolution coupled with substantial cation exchange. Conversely, the water in the P1t coalbed is mainly influenced by silicate weathering/dissolution as well as evaporative concentration, with a limited occurrence of cation exchange. Moreover, there are distinct disparities in ion sources between P1s and P1t. These research findings provide a scientific foundation for assessing the development potential of CBM and optimizing extraction systems within similar CBM areas.

Unlocking the potential of N-acetylcysteine: Improving hepatopancreas inflammation, antioxidant capacity and health in common carp (Cyprinus carpio) via the MAPK/NF-κB/Nrf2 signalling pathway.

N-acetylcysteine (NAC) positively contributes to enhancing animal health, regulating inflammation and reducing stress by participating in the synthesis of cysteine, glutathione, and taurine in the body. The present study aims to investigate the effects of dietary different levels of NAC on the morphology, function and physiological state of hepatopancreas in juvenile common carp (Cyprinus carpio). 450 common carps were randomly divided into 5 groups: N1 (basal diet), N2 (1.5 g/kg NAC diet), N3 (3.0 g/kg NAC diet), N4 (4.5 g/kg NAC diet) and N5 (6.0 g/kg NAC diet), and fed for 8 weeks. The results indicated that dietary 3.0-6.0 g/kg NAC reduced hepatopancreas lipid vacuoles and nuclear translocation, and inhibited apoptosis in common carp. Simultaneously, the activities of hepatopancreas alanine aminotransferase and aspartate aminotransferase progressively increased with rising dietary NAC levels. Dietary NAC enhanced the non-specific immune function of common carp, and exerted anti-inflammatory effects by inhibiting the MAPK/NF-κB signaling pathway. Additionally, dietary 3.0-6.0 g/kg NAC significantly improved the antioxidant capacity of common carp, which was associated with enhanced glutathione metabolism, clearance of ROS and the activation of Nrf2 signaling pathway. In summary, NAC has the potential to alleviate inflammation, mitigate oxidative stress and inhibit apoptosis via the MAPK/NF-κB/Nrf2 signaling pathway, thereby improving hepatopancreas function and health of common carp. The current findings provide a theoretical basis for promoting the application of NAC in aquaculture and ecological cultivation of aquatic animals.

Application of 3.0 T multimodal MRI in FIGO staging of cervical cancer.

BACKGROUND: In malignant tumours of the female reproductive system, cervical cancer is second only to breast cancer, seriously threatening the health and safety of most women. OBJECTIVE: To evaluate the clinical value of 3.0 T multimodal nuclear magnetic resonance imaging (MRI) in the International Federation of Gynecology and Obstetrics' (FIGO) staging of cervical cancer. METHODS: The clinical data of 30 patients with pathologically diagnosed cervical cancer admitted to our hospital from January 2018 to August 2022 were analysed retrospectively. Before treatment, all patients were examined with conventional MRI, diffusion-weighted imaging and multi-directional contrast-enhanced imaging. RESULTS: The accuracy of multimodal MRI in the FIGO staging of cervical cancer (29/30, 96.7%) was significantly higher than the accuracy obtained in a control group (21/30, 70.0%), with a statistically significant difference (p= 0.013). In addition, there was good agreement between two observers applying multimodal imaging (kappa= 0.881) and moderate agreement between two observers in the control group (kappa= 0.538). CONCLUSION: Multimodal MRI can evaluate cervical cancer comprehensively and accurately to enable accurate FIGO staging, providing significant evidence for clinical operation planning and subsequent combined therapy.

Effects of hydroxy methionine zinc on growth performance, immune response, antioxidant capacity, and intestinal microbiota of red claw crayfish (Procambarus clarkii).

This study aimed to evaluate the effects of varying zinc (Zn) levels on the growth performance, non-specific immune response, antioxidant capacity, and intestinal microbiota of red claw crayfish (Procambarus clarkii (P. clarkii)). Adopting hydroxy methionine zinc (Zn-MHA) as the Zn source, 180 healthy crayfish with an initial body mass of 6.50 ± 0.05 g were randomly divided into the following five groups: X1 (control group) and groups X2, X3, X4, and X5, which were fed the basal feed supplemented with Zn-MHA with 0, 15, 30, 60, and 90 mg kg-1, respectively. The results indicated that following the addition of various concentrations of Zn-MHA to the diet, the following was observed: Specific growth rate (SGR), weight gain rate (WGR), total protein (TP), total cholesterol (TC), the activities of alkaline phosphatase (AKP), phenoloxidase (PO), total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD) and catalase (CAT), the expression of CTL, GPX, and CuZn-SOD genes demonstrated a trend of rising and then declining-with a maximum value in group X4-which was significantly higher than that in group X1 (P < 0.05). Zn deposition in the intestine and hepatopancreas, the activity of GSH-PX, and the expression of GSH-PX were increased, exhibiting the highest value in group X5. The malonaldehyde (MDA) content was significantly reduced, with the lowest value in group X4, and the MDA content of the Zn-MHA addition groups were significantly lower than the control group (P < 0.05). In the analysis of the intestinal microbiota of P. clarkii, the number of operational taxonomic units in group X4 was the highest, and the richness and diversity indexes of groups X3 and X4 were significantly higher than those in group X1 (P < 0.05). Meanwhile, the dietary addition of Zn-MHA decreased and increased the relative abundance of Proteobacteria and Tenericutes, respectively. These findings indicate that supplementation of dietary Zn-MHA at an optimum dose of 60 mg kg-1 may effectively improve growth performance, immune response, antioxidant capacity, and intestinal microbiota richness and species diversity in crayfish.

Evidence of Coal Biodegradation from Coalbed-Produced Water - A Case Study of Dafosi Gas Field, Ordos Basin, China.

Bioconversion of coal to methane occurs in the coalbed aquifer environment. To investigate the evidence of coal biodegradation from coalbed-produced water, we collected six field water samples from the Dafosi gas field and prepared one laboratory-simulated water sample and one indoor anaerobic microbial degradation sample with the highest compound concentration as the two reference standards. Gas chromatography-mass spectrometry was used to detect the organic compound type, concentration, and differences in the biomarker compound sensitivity. Results indicate that extracted organic matter from coalbed-produced water samples can be evidence of biodegradation. Variations in range compounds (such as n-alkanes, tri- and pentacyclic terpenes, and steranes) and their sensitivity confirmed active microbial degradation in the studied area. A positive correlation between the n-alkanes content in the coalbed-produced water and the stable carbon isotope value of methane further verifies that the n-alkanes are primary substrates for maintaining microbial activity. Therefore, evidence including n-alkanes, tri- and pentacyclic terpenes, steranes, unresolved complex mixtures, and stable carbon isotope composition of methane contribute to biogenic methane generation in situ. Our limited data suggest that managing soluble organic matter in the coalbed-produced water may provide a viable route for coal biodegradation since most microorganisms survive within the coal seam water.

The Association of the Triglyceride and Muscle to Fat Ratio During Early Pregnancy with the Development of Gestational Diabetes Mellitus.

Objective: This study explored the association between metabolic factors and body composition during the first trimester of gestational diabetes mellitus (GDM). Methods: This prospective study recruited pregnant women in their first trimester. Clinical information and glucose and lipid measurements were collected, and body composition was assessed using multifrequency bioelectrical impedance analysis. GDM was diagnosed on the basis of an oral glucose tolerance test at 24-28 gestational week. Factors related to GDM were investigated using correlation, and risk ratios (RRs) and 95% CIs of potential risk factors with GDM were estimated using Poisson regression. The area under the receiver operating characteristic (ROC) curve was used to determine predictive effects. Results: 59/302 women (19.5%) developed GDM. Older (RR 1.076, 95% CI 1.005-1.152), higher body mass index (BMI) before pregnancy (pre-BMI) (RR 1.012, 95% CI 1.005-1.063), triglycerides (RR 4.052, 95% CI 1.641-6.741), and lower skeletal muscle mass (SMM) to fat mass (FM) ratio (SMM/FM) (RR 0.213, 95% CI 0.051-0890) in the first trimester, and family history of type 2 diabetes (RR 1.496, 95% CI 1.014-2.667) significantly associated with the risk of GDM, but neither fasting plasma glucose nor glycated albumin was associated with GDM. The combined multivariate prediction model achieved good discrimination with an AUC of 0.806 (95% CI 0.737-0.895, P<0.001). According to ROC curve, the cut-off values of TG and SMM/FM were 0.925 mmol/L and 1.305. Conclusion: Reduced SMM/FM and elevated triglyceride (TG) levels in the first trimester are associated with GDM development, and should be screened in early pregnancy to identify high-risk subjects for GDM.

Government regulation strategy, leading firms' innovation strategy, and following firms imitation strategy: An analysis based on evolutionary game theory.

In the innovation ecosystem, the knowledge-based game behavior of each subject not only pertains to its own survival and development but also affects evolution of the innovation ecosystem. The present study investigates the choice of government's regulation strategy, leading firms' innovation protection strategy and following firms' imitation strategy from the perspective of group evolutionary game. Based on the cost-benefit perspective, an asymmetric tripartite evolutionary game model and a simulation model are constructed to analyze the strategies and stability of the evolutionary equilibrium of each subject. We focus mainly on the protection intensity of innovation achievements by leading enterprises and the difficulty of imitation and substitution by following enterprises. The cost of patent operation and maintenance, government subsidies, and the relative difficulty of technology substitution and imitation were identified as the key factors affecting the evolutionary equilibrium of the system. Based on different scenarios resulting from the aforementioned factors, four equilibrium states are observed in the system, namely {no government regulation, technology secrecy, substitution}, {no government regulation, technology secrecy, imitation}, {no government regulation, patent application, imitation}, and {government regulation, patent application, imitation}. Finally, the study suggests corresponding recommendations for the three parties, which can help governments as well as the leading and following firms to choose appropriate behavioral strategies. At the same time, this study offers positive insights to participants in the global innovation ecosystem.

The Effect of Initial Texture on the Plastic Deformation of Gradient Aluminum.

The effect of specific processing-induced surface textures in gradient aluminum has not yet been investigated. A dislocation-based multi-scale framework is employed to explore the influence of various initial shearing textures and the depth from the surface of the region featuring each texture on the macroscopic behavior of gradient aluminum. By assigning different textures to the same grain size gradient aluminum sample, the initial texture was found to significantly affect the plastic deformation and macroscopic behavior of gradient aluminum. Specifically, the {111} texture can enhance the strength-ductility synergy, and this effect is dependent on the depth from the surface where the texture is located. This texture can lead to a slow stress/strain gradient in the assigned texture region and a sharp stress/strain gradient in the grain size gradient region connecting this region with the coarse grain region. Particularly, the sharp stress/strain gradient can result in extra strengthening by adjusting the stress/strain localization. These findings provide valuable insights for the design and optimization of surface textures in gradient aluminum.

Comparative Study on Growth Index and Nutritional Quality of Female Chinese Mitten Crab Eriocheir sinensis Selected at Different Growth Periods in Rice-Crab Culture Systems.

Research was conducted on the growth performance and nutritional quality of Chinese mitten crabs (Eriocheir sinensis) during a 62-day growing period in a symbiotic coculture comprising rice and crab. Culture experiments were conducted in three rice fields of equal size (996 m2). On days 0 (July 15, D0), 15 (July 30, D15), 31 (August 15, D31), 46 (August 30, D46), and 62 (September 2, D62), tissue samples of 50 female E. sinensis were collected randomly from each rice field. The results showed that the serum growth hormone (GH) content and muscle ecdysone receptor (EcR) mRNA expression levels were higher in the D31 and D46 groups; the content of serum 20-hydroxyecdysone (20-HE) and the mRNA expression levels of retinoid X receptor (RXR), insulin-like growth factor 2 (IGF2), and chitinase (CHI) reached the maximum in the D31 group. Muscle crude protein content gradually increased; hepatopancreas crude protein and crude lipid content began to decrease after reaching the maximum value in the D0 and D15 groups, respectively; the contents of crude protein and crude lipid in the ovary significantly increased in the D46 and D62 groups (P < 0.05). The content of muscle essential amino acids (EAA) reached the maximum in the D46 group; the hepatopancreas EAA content began to decrease significantly in the D31 group (P < 0.05); and the EAA content of the ovary decreased significantly after reaching the maximum value in the D46 group (P < 0.05). The muscle contents of eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), docosahexaenoic acid (DHA), polyunsaturated fatty acids (PUFA), and n-3 polyunsaturated fatty acids (n-3PUFA) and the ratio of n-3 polyunsaturated fatty acids/n-6 polyunsaturated fatty acids (n3/n6) decreased significantly in the D31 group (P < 0.05); the hepatopancreas contents of EPA, PUFA, n-3PUFA, and n-6 polyunsaturated fatty acids (n-6PUFA) and the ratio of n3/n6 began to decrease after reaching the maximum value in the D31 group, ethyl behenate (21:0), tetracosanoic acid (24:0), DPA, and DHA contents were detected for the first time in the D31 group; the ovary PUFA, n-3PUFA contents, and n3/n6 ratio of the D46 and D62 groups were significantly lower than those of the D31 group (P < 0.05). During the experimental conditions described here, female E. sinensis raised in rice fields reached rapid growth from August 15 to August 30. Additionally, the nutritional quality of the female E. sinensis edible tissues (muscle, hepatopancreas, and ovary) began to decline after August 15, when sufficient nutrients such as protein, lipid, EAA, and PUFA should be provided to the female E. sinensis.

Effects of opioid-free propofol or remimazolam balanced anesthesia on hypoxemia incidence in patients with obesity during gastrointestinal endoscopy: A prospective, randomized clinical trial.

There are presently no consensuses on the optimal sedation strategy for obese patients during gastrointestinal endoscopy. This study aim to explore the effects of opioid-free propofol or remimazolam balanced anesthesia on hypoxemia incidence in patients with obesity. A total of 264 patients were randomized to remimazolam + esketamine group (group R) or propofol + esketamine group (group P). Anesthesia in group P was administrated by propofol, esketamine and in group R by remimazolam, esketamine. The primary outcome was incidence of hypoxemia. Secondary outcomes were the time to loss of consciousness (LoC) and to recovery and the incidence of intraoperative and postoperative adverse reactions. We found the incidence of mild hypoxemia in group R was similar to that in group P (14.2% vs. 11.5%, p = 0.396). The incidence of severe hypoxemia in group R was significantly lower than Group P (4.2% vs. 9.2%, p = 0.019). The time to LoC in group R was longer than group P [Median (interquartile range, IQR): 53 s (45 to 61) vs. 50 s (42 to 54), p = 0.001]. The time to recovery from anesthesia in group R was less than group P [Median (IQR): 48 min (41 to 58) vs. 55.5 min (46 to 67), p<0.001]. There was no significant difference in the incidence of adverse events (p > 0.05 for all). We concluded that compared with propofol combined with esketamine, remimazolam combined with esketamine can reduce the incidence of severe hypoxemia during gastrointestinal endoscopy in obese patients. Clinical Trial Registration: www.chictr.org.cn, Identifier: ChiCTR2200065575.

Heading perception from optic flow occurs at both perceptual representation and working memory stages with EEG evidence.

Psychophysical studies have demonstrated that heading perception from optic flow occurs in perceptual and post-perceptual stages. The post-perception stage is a complex concept, containing working memory. The current study examined whether working memory was involved in heading perception from optic flow by asking participants to conduct a heading perception task and recording their scalp EEG. On each trial, an optic flow display was presented, followed by a blank display. Participants were then asked to report their perceived heading. We know that participants would tend to automatically forget previous headings when they learned that previously presented headings were unrelated to the current heading perception to save cognitive resources. As a result, we could not decode previous headings from the EEG data of current trials. More importantly, if we successfully decoded previous headings when the blank display (optic flow) was presented, then working memory (perceptual representation stage) was involved in heading perception. Our results showed that the decoding accuracy was significantly higher than the chance level when the optic flow and blank displays were presented. Therefore, the current study provided electrophysiological evidence that heading perception from optic flow occurred in the perceptual representation and working memory stages, against the previous perceptual claim.

Recent progress in metabolic reprogramming in gestational diabetes mellitus: a review.

Gestational diabetes mellitus is a prevalent metabolic disease that can impact the normal course of pregnancy and delivery, leading to adverse outcomes for both mother and child. Its pathogenesis is complex and involves various factors, such as insulin resistance and ß-cell dysfunction. Metabolic reprogramming, which involves mitochondrial oxidative phosphorylation and glycolysis, is crucial for maintaining human metabolic balance and is involved in the pathogenesis and progression of gestational diabetes mellitus. However, research on the link and metabolic pathways between metabolic reprogramming and gestational diabetes mellitus is limited. Therefore, we reviewed the relationship between metabolic reprogramming and gestational diabetes mellitus to provide new therapeutic strategies for maternal health during pregnancy and reduce the risk of developing gestational diabetes mellitus.

First deliveries of felines by transcervical transfer of in vitro-cultured embryos.

Transcervical transplantation of embryos has been wildly used in several species. However, it has not been successful in feline species because of their unique cervix structure combined with poor in vitro culture systems for embryos. In the present study, a simple device to guide the embryo transfer (ET) tube through the cervix and into the uterus was developed. To supply better blastocysts for ET, the in vitro culture system of feline embryos was also optimized. Results showed that 91.67 ± 4.17% of embryos developed to the blastocyst stage in QAU-4 medium compared with 49.17 ± 6.51% in KSOM medium. The total cell number of blastocysts reached 87.1 ± 22.1 in QAU-4 medium compared with 29.8 ± 2.1 in KSOM. Embryoid body formation was also observed in QAU-4 medium. Finally, by the improved technical scheme, CRX (Cone-Rod Homeobox) gene-edited kittens were successfully bred. To date, this is the first report, to our knowledge, of knockout kittens successfully delivered after transcervical ET with a simple transfer tube.