OBJECTIVE: To explore the serological and molecular genetic characteristics of a family with subtype B(A)06. METHODS: A neonatal hyperbilirubinemia patient who was treated at Henan Children's Hospital on June 15, 2023 due to "yellowing of the skin and gradual aggravation", and was found to have inconsistent ABO forward and reverse typing through blood type testing, was selected as the research subject. Six milliliters of peripheral blood were collected from the newborn and her family members (grandfather, grandmother, father, mother and aunt) respectively. ABO blood group identification was performed by the blood group serological method. Human genomic DNA was extracted using the nucleic acid extraction or purification reagent BT-01. ABO gene exons 2 to 7 were amplified by PCR. The PCR-specific products that were successfully amplified were sequenced by Sanger method. Taking ABO*A1.01 as the reference sequence, the ABO gene sequences of the newborn and her family members were analyzed to determine the ABO genotype. The procedures followed in this study were approved by the Ethics Committee of Henan Children's Hospital (Ethics No.: 2022-K-L036). RESULTS: The serological results of ABO blood group showed that the newborn, her grandfather, father and aunt were all incompatible with the forward and reverse typing. The blood group phenotype of the newborn was AwB or B(A), the blood group phenotype of the grandfather was A2B or B(A), the blood group phenotype of the father and aunt were A2B, and the blood group phenotype of the grandmother and mother were both O. The screening test results of hemolytic disease of the newborn showed that the free test detected IgG anti-A1 antibody, while the elution test, direct antiglobulin test and antibody screening results were all negative. The Sanger sequencing results showed that the newborn had variations of c.261delG, c.297A>G, c.526C>G, c.657C>T, c.703G>A, c.796C>A and c.930G>A. Her grandfather had variations of c.297A>G, C.526C>G, c.657C>T, c.703G>A, c.796C>A, c.803G>C and c.930G>A. Her grandmother had variations of c.106G>T, c.188G>A, c.189C>T, c.220C>T, c.261delG, c.297A>G, c.646T>A, c.681G>A, c.771C>T and c.829G>A. Her father and aunt had variations of c.106G>T, c.188G>A, c.189C>T, c.220C>T, c.261delG, c.297A>G, c.526C>G, c.646T>A, c.657C>T, c.681G>A, c.703G>A, c.771C>T, c.796C>A, c.829G>A and c.930G>A. Her mother had variations of c.106G>T, c.188G>A, c.189C>T, c.220C>T, c.261delG, c.297A>G, c.646T>A, c.681G>A, c.771C>T, and c.829G>A.The genotype of the newborn was ABO*BA.06/ABO*O.01.01, her grandfather was ABO*BA.06/ABO*B.01, her grandmother was ABO*O.01.02/ABO*O.01.02, her father and aunt were ABO*BA.06/ABO*O.01.02, and her mother was ABO*O.01.01/ABO*O.01.02. The ABO*BA.06 allele of the newborn, grandfather, father and aunt was caused by the c.803C>G variation in exon 7 based on the ABO*B.01 allele. The ABO*BA.06 allele can be stably inherited in this family. CONCLUSION The blood type of neonatal patients with B(A)06 subtype can be accurately determined by gene sequencing technology. If the forward typing is ≤ 3+ agglutination intensity in newborn ABO blood group identification, the reason should be carefully analyzed, and the molecular biology technology and family gene sequencing results should be used to jointly determine if necessary.
Humans ; ABO Blood-Group System/genetics* ; Female ; Pedigree ; Male ; Infant, Newborn ; Asian People/genetics* ; Genotype ; China ; Blood Grouping and Crossmatching ; Hyperbilirubinemia, Neonatal/blood* ; East Asian People

BACKGROUND:Dexamethasone and hindlimb suspension are commonly used methods for modeling sarcopenia in animal experiments due to their short modeling time,ease of operation,and low cost.OBJECTIVE:To compare the differences in muscle mass,strength and functional phenotypes and molecular mechanisms between two mouse sarcopenia models induced by dexamethasone and hindlimb suspension.METHODS:Thirty male C57BL/6 mice were randomly divided into three groups(n=10 per group).The normal control group received no intervention.The dexamethasone group received daily intraperitoneal injections of 1 mg/kg/d dexamethasone sodium phosphate solution for 6 continuous days to establish sarcopenia models in mice,while mice in the hindlimb suspension group were suspended by tail harness for 16 hours,once per day,to establish sarcopenia models.Within 6 weeks after modeling,changes in body mass were monitored.After 6 weeks of modeling,mice were tested for limb grip strength,mobility(swimming test),skeletal muscle wet mass,and skeletal muscle pathological morphology.Expressions of skeletal muscle protein synthesis and catabolism indexes as well as the AMPK/FoXO3α signaling pathway were detected by RT-PCR and western blot.RESULTS AND CONCLUSION:(1)Two weeks after modeling,both dexamethasone and hindlimb suspension groups showed a significant decrease in body mass compared with the normal control group(P<0.001).After 6 weeks of modeling,grip strength of mice in both dexamethasone and hindlimb suspension groups was lower than that in the normal control group(P<0.001).The wet mass of gastrocnemius and extensor digitorum longus muscles and the cross-sectional area of gastrocnemius and soleus muscles in the dexamethasone group were lower than those in the normal control group(P<0.05).Compared with the hindlimb suspension group,the cross-sectional area of gastrocnemius muscle was significantly smaller in the dexamethasone group(P<0.05),while the cross-sectional area of soleus muscle was larger in the dexamethasone group(P<0.05).Mice in the dexamethasone group had reduced mobility when compared with those in the normal control group and the hindlimb suspension group(P<0.05).(3)Compared with the normal control group,PI3K,mTOR,AMPK,and PGC-1α mRNA expression and P-AMPK/AMPK protein were decreased in the two modeling groups(P<0.05),and FoXO3α mRNA expression and PGC-1α and FoXO3 protein expression were elevated(P<0.05);in the dexamethasone group,Akt1 mRNA expression was decreased(P<0.05),while Atrogin-1 and MuRF-1 mRNA expression was elevated(P<0.05);in the hindlimb suspension group,Akt1 mRNA expression was elevated(P<0.05).(4)Compared with the dexamethasone group,mTOR,Akt1,and FoXO3α mRNA expression was elevated in the hindlimb suspension group(P<0.05),while Atrogin-1 and MuRF-1 mRNA expression was decreased(P<0.05).To conclude,both modeling methods could decrease the levels of mitochondrial energy metabolism in skeletal muscle,with the dexamethasone group mediating atrophy of skeletal muscle through the dual action of ubiquitin proteasome and energy metabolism pathways,and the hindlimb suspension group inducing atrophy of skeletal muscle by mediating the energy metabolism pathway through the AMPK/FoXO3α signaling pathway,subsequently causing a reduction in mass,strength,and function of skeletal muscle.

BACKGROUND:Dexamethasone and hindlimb suspension are commonly used methods for modeling sarcopenia in animal experiments due to their short modeling time,ease of operation,and low cost.OBJECTIVE:To compare the differences in muscle mass,strength and functional phenotypes and molecular mechanisms between two mouse sarcopenia models induced by dexamethasone and hindlimb suspension.METHODS:Thirty male C57BL/6 mice were randomly divided into three groups(n=10 per group).The normal control group received no intervention.The dexamethasone group received daily intraperitoneal injections of 1 mg/kg/d dexamethasone sodium phosphate solution for 6 continuous days to establish sarcopenia models in mice,while mice in the hindlimb suspension group were suspended by tail harness for 16 hours,once per day,to establish sarcopenia models.Within 6 weeks after modeling,changes in body mass were monitored.After 6 weeks of modeling,mice were tested for limb grip strength,mobility(swimming test),skeletal muscle wet mass,and skeletal muscle pathological morphology.Expressions of skeletal muscle protein synthesis and catabolism indexes as well as the AMPK/FoXO3α signaling pathway were detected by RT-PCR and western blot.RESULTS AND CONCLUSION:(1)Two weeks after modeling,both dexamethasone and hindlimb suspension groups showed a significant decrease in body mass compared with the normal control group(P<0.001).After 6 weeks of modeling,grip strength of mice in both dexamethasone and hindlimb suspension groups was lower than that in the normal control group(P<0.001).The wet mass of gastrocnemius and extensor digitorum longus muscles and the cross-sectional area of gastrocnemius and soleus muscles in the dexamethasone group were lower than those in the normal control group(P<0.05).Compared with the hindlimb suspension group,the cross-sectional area of gastrocnemius muscle was significantly smaller in the dexamethasone group(P<0.05),while the cross-sectional area of soleus muscle was larger in the dexamethasone group(P<0.05).Mice in the dexamethasone group had reduced mobility when compared with those in the normal control group and the hindlimb suspension group(P<0.05).(3)Compared with the normal control group,PI3K,mTOR,AMPK,and PGC-1α mRNA expression and P-AMPK/AMPK protein were decreased in the two modeling groups(P<0.05),and FoXO3α mRNA expression and PGC-1α and FoXO3 protein expression were elevated(P<0.05);in the dexamethasone group,Akt1 mRNA expression was decreased(P<0.05),while Atrogin-1 and MuRF-1 mRNA expression was elevated(P<0.05);in the hindlimb suspension group,Akt1 mRNA expression was elevated(P<0.05).(4)Compared with the dexamethasone group,mTOR,Akt1,and FoXO3α mRNA expression was elevated in the hindlimb suspension group(P<0.05),while Atrogin-1 and MuRF-1 mRNA expression was decreased(P<0.05).To conclude,both modeling methods could decrease the levels of mitochondrial energy metabolism in skeletal muscle,with the dexamethasone group mediating atrophy of skeletal muscle through the dual action of ubiquitin proteasome and energy metabolism pathways,and the hindlimb suspension group inducing atrophy of skeletal muscle by mediating the energy metabolism pathway through the AMPK/FoXO3α signaling pathway,subsequently causing a reduction in mass,strength,and function of skeletal muscle.

Objective:To propose suggestions for the revision of quality standard of steviol glycosides through research on its major manufacturer's products.Methods:The inspection items in the current quality standards were inspected,with domestic and international quality standard of steviol glycosides.Results:Specific rotation was from-33.39° to-39.50°,pH was from 5.3 to 6.7,impurity absorbency was lower than 0.096 0,while the contents of weight loss on drying were lower than 5.0％,residue on ignition were lower than 0.043％,methanol residual were lower than 0.01％,ethanol residual were lower than 0.06％,residue on arsenic were lower than 0.000 054％,lead wasn't detected.The content of rebaudioside A and 13 kinds of steviol glycosides were 8.23％-68.48％and 85.78％-96.13％.Conclusion:The main components of the commercial steviol glycosides products were rebaudioside A and stevioside,with low safety risk of elemental impurities and solvent residues.It provided suggestions for the revision of quality standard of steviol glycosides in the Chinese Pharmacopoeia.

Objective:To propose suggestions for the revision of quality standard of steviol glycosides through research on its major manufacturer's products.Methods:The inspection items in the current quality standards were inspected,with domestic and international quality standard of steviol glycosides.Results:Specific rotation was from-33.39° to-39.50°,pH was from 5.3 to 6.7,impurity absorbency was lower than 0.096 0,while the contents of weight loss on drying were lower than 5.0％,residue on ignition were lower than 0.043％,methanol residual were lower than 0.01％,ethanol residual were lower than 0.06％,residue on arsenic were lower than 0.000 054％,lead wasn't detected.The content of rebaudioside A and 13 kinds of steviol glycosides were 8.23％-68.48％and 85.78％-96.13％.Conclusion:The main components of the commercial steviol glycosides products were rebaudioside A and stevioside,with low safety risk of elemental impurities and solvent residues.It provided suggestions for the revision of quality standard of steviol glycosides in the Chinese Pharmacopoeia.

OBJECTIVE To investigate the role of Bai Ling Long Zao An Shen formula(BLLZ)in sleep improvement in an environmental stress-induced insomnia rat model and explore its underlying mechanisms.METHODS(1)Component analysis:the chemical constituents of the BLLZ extract were analyzed using ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS).(2)Eval-uation of the sedative and hypnotic effect:① Mice:50 ICR mice were randomly divided into normal control group,BLLZ-L group(5,10 and 20 g·kg-1)and diazepam group(DZP,3 mg·kg-1).After five days of intragastric administration,pentobarbital sodium-induced righting reflex and locomotor activity tests were performed.② Rats:8 SD rats were implanted with electrodes and allowed to recover for seven days before baseline EEG data was collected over 24 h.A crossover design(7 d washout period)was employed,with rats randomly assigned to the DZP(3 mg·kg-1)and BLLZ(20 g·kg-1)group.After five days of treatment,24 h EEG recordings were obtained.(3)Insomnia model and interventions:①8 SD rats were allowed to recover for seven days post-surgery,followed by 6 h(14:00-20:00)baseline EEG recording.A 3×3 crossover design was used to assign rats to model(environmental stress-induced insomnia),model+DZP,or model+BLLZ groups.After five days of treatment,insomnia was induced by frequent cage changes(14:00,16:00 and 18:00),and EEG changes were monitored.(4)Mechanistic study:32 SD rats were randomly divided into the normal control group,model group,and model+DZP group.After five days of treatment,hypothalamic tissues were collected for biochemi-cal analysis.γ-aminobutyric acid(GABA),glutamate(Glu),and dopamine(DA)levels were measured using biochemical kits while γ aminobutyric acid receptor subunit alpha-1(GABAA1),core clock proteins period circadian regulator 2(PER2)and circadian locomotor output cycles(CLOCK)protein expressions were assessed by Western blotting.RESULTS(1)Compared with the normal control group,the sleep latency of BLLZ 10 and 20 g·kg-1 and DZP groups was significantly shortened,and the locomotor activity of BLLZ 20 g·kg-1 and DZP groups was significantly reduced;BLLZ 20 g·kg-1 signifi-cantly increased the total sleep time,slow-wave sleep time,and average duration of sleep in normal rats,and significantly reduced the wakefulness time.(2)The total sleep time and slow-wave sleep time of the model group significantly decreased and the wakefulness time significantly increased compared with baseline.(3)Compared with the model group,the total sleep time and slow-wave sleep time of the model+BLLZ group and the model+DZP group were significantly increased,and the wakefulness time significantly shortened.(4)Compared with the normal control group,the Glu/GABA ratio,DA content and CLOCK protein expression were significantly increased and GABAA1 and PER2 protein expres-sion were significantly decreased in the model group;compared with the model group,the Glu/GABA ratio,DA content and CLOCK protein expression were significantly decreased,and the expression of GABAA1 and PER2 were significantly increased in the model+BLLLZ group and the model+DZP group.CONCLUSION BLLZ has sedative and hypnotic effects.It can prolong the total slow-wave sleep time by increasing the average duration of slow-wave sleep episodes,thereby increasing the total sleep time and improving environmental stress-induced insomnia.The mechanism may be related to the downregulation of the Glu/GABA ratio and DA levels as well as the enhancement of GABAA1 expressions and the regulation of hypothalamic core clock protein expressions.

Objective:To explore the safety and effectiveness of ileal bypass with stent to prevent anastomotic leakage in Da Vinci robot anus-preserving surgery for low rectal cancer.Methods:From Dec 2021 to Sep 2024, the clinical data of 55 patients undergoing robot anus-preserving surgery and stent ileal bypass for low rectal cancer in the First Affiliated Hospital of Ningbo University was analyzed retrospectively.Results:All the 55 patients successfully completed the operation. Among them, 47 patients underwent low anterior resection and 8 patients did sphincter resection. The total operation time was (207±37)min, the operation time of ileal bypass was (24±3)min, and the intraoperative blood loss was (32±18)ml. After operation, the time of stent disintegration was (23.93± 2.87) days, and the time of catheter removal was (29.55 ± 3.21) days. By postoperative TNM staging, there were 12 cases in T1 stage, 26 cases in T2 and 17 cases in T3. The number of lymph node dissection was (16.84±4.50), 15 cases being positive for lymph node metastasis, 40 cases being negtive for lymph node metastasis, 36 cases were stage Ⅰ, 4 cases were stage Ⅱ and 15 cases were stage Ⅲ. The median follow-up time was 20 (2-35) months. Delayed anastomotic leakage occurred in 1 patient and anastomotic stenosis occurred in 2 patients.Conclusion:Stent ileal bypass is safe and effective to prevent anastomotic leakage in Da Vinci robot anus-preserving surgery for low rectal cancer.

OBJECTIVE To investigate the role of Bai Ling Long Zao An Shen formula(BLLZ)in sleep improvement in an environmental stress-induced insomnia rat model and explore its underlying mechanisms.METHODS(1)Component analysis:the chemical constituents of the BLLZ extract were analyzed using ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS).(2)Eval-uation of the sedative and hypnotic effect:① Mice:50 ICR mice were randomly divided into normal control group,BLLZ-L group(5,10 and 20 g·kg-1)and diazepam group(DZP,3 mg·kg-1).After five days of intragastric administration,pentobarbital sodium-induced righting reflex and locomotor activity tests were performed.② Rats:8 SD rats were implanted with electrodes and allowed to recover for seven days before baseline EEG data was collected over 24 h.A crossover design(7 d washout period)was employed,with rats randomly assigned to the DZP(3 mg·kg-1)and BLLZ(20 g·kg-1)group.After five days of treatment,24 h EEG recordings were obtained.(3)Insomnia model and interventions:①8 SD rats were allowed to recover for seven days post-surgery,followed by 6 h(14:00-20:00)baseline EEG recording.A 3×3 crossover design was used to assign rats to model(environmental stress-induced insomnia),model+DZP,or model+BLLZ groups.After five days of treatment,insomnia was induced by frequent cage changes(14:00,16:00 and 18:00),and EEG changes were monitored.(4)Mechanistic study:32 SD rats were randomly divided into the normal control group,model group,and model+DZP group.After five days of treatment,hypothalamic tissues were collected for biochemi-cal analysis.γ-aminobutyric acid(GABA),glutamate(Glu),and dopamine(DA)levels were measured using biochemical kits while γ aminobutyric acid receptor subunit alpha-1(GABAA1),core clock proteins period circadian regulator 2(PER2)and circadian locomotor output cycles(CLOCK)protein expressions were assessed by Western blotting.RESULTS(1)Compared with the normal control group,the sleep latency of BLLZ 10 and 20 g·kg-1 and DZP groups was significantly shortened,and the locomotor activity of BLLZ 20 g·kg-1 and DZP groups was significantly reduced;BLLZ 20 g·kg-1 signifi-cantly increased the total sleep time,slow-wave sleep time,and average duration of sleep in normal rats,and significantly reduced the wakefulness time.(2)The total sleep time and slow-wave sleep time of the model group significantly decreased and the wakefulness time significantly increased compared with baseline.(3)Compared with the model group,the total sleep time and slow-wave sleep time of the model+BLLZ group and the model+DZP group were significantly increased,and the wakefulness time significantly shortened.(4)Compared with the normal control group,the Glu/GABA ratio,DA content and CLOCK protein expression were significantly increased and GABAA1 and PER2 protein expres-sion were significantly decreased in the model group;compared with the model group,the Glu/GABA ratio,DA content and CLOCK protein expression were significantly decreased,and the expression of GABAA1 and PER2 were significantly increased in the model+BLLLZ group and the model+DZP group.CONCLUSION BLLZ has sedative and hypnotic effects.It can prolong the total slow-wave sleep time by increasing the average duration of slow-wave sleep episodes,thereby increasing the total sleep time and improving environmental stress-induced insomnia.The mechanism may be related to the downregulation of the Glu/GABA ratio and DA levels as well as the enhancement of GABAA1 expressions and the regulation of hypothalamic core clock protein expressions.

Drug repurposing offers a promising alternative to traditional drug development and significantly reduces costs and timelines by identifying new therapeutic uses for existing drugs. However, the current approaches often rely on limited data sources and simplistic hypotheses, which restrict their ability to capture the multi-faceted nature of biological systems. This study introduces adaptive multi-view learning (AMVL), a novel methodology that integrates chemical-induced transcriptional profiles (CTPs), knowledge graph (KG) embeddings, and large language model (LLM) representations, to enhance drug repurposing predictions. AMVL incorporates an innovative similarity matrix expansion strategy and leverages multi-view learning (MVL), matrix factorization, and ensemble optimization techniques to integrate heterogeneous multi-source data. Comprehensive evaluations on benchmark datasets (Fdataset, Cdataset, and Ydataset) and the large-scale iDrug dataset demonstrate that AMVL outperforms state-of-the-art (SOTA) methods, achieving superior accuracy in predicting drug-disease associations across multiple metrics. Literature-based validation further confirmed the model's predictive capabilities, with seven out of the top ten predictions corroborated by post-2011 evidence. To promote transparency and reproducibility, all data and codes used in this study were open-sourced, providing resources for processing CTPs, KG, and LLM-based similarity calculations, along with the complete AMVL algorithm and benchmarking procedures. By unifying diverse data modalities, AMVL offers a robust and scalable solution for accelerating drug discovery, fostering advancements in translational medicine and integrating multi-omics data. We aim to inspire further innovations in multi-source data integration and support the development of more precise and efficient strategies for advancing drug discovery and translational medicine.

Objective To investigate the relationship between the levels of serum suppressor of cytokine signaling 3(SOCS3),growth differentiation factor-15(GDF-15)and liver fibrosis in patients with type 2 dia-betes mellitus(T2DM)combined with non-alcoholic fatty liver disease(NAFLD).Methods A total of 320 patients with T2DM combined with NAFLD who were hospitalized in this hospital from May 2023 to May 2024 were selected as the study group,and another 320 patients with simple T2DM admitted during the same period were selected as the control group.The levels of serum SOCS3 and GDF-15 were determined by en-zyme-linked immunosorbent assay(ELISA).Pearson correlation analysis was used to analyze the correlation between SOCS3,GDF-15 levels and liver fibrosis indicators.Logistic regression analysis was used to analyze the factors affecting the degree of liver fibrosis.The receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of serum SOCS3 and GDF-15 for the degree of liver fibrosis in patients.Results Compared with the control group,the levels of serum SOCS3,GDF-15,5,type Ⅲ procollagen peptide,laminin and type Ⅳ col-lagen in the study group were significantly increased(P＜0.05).There were statistically significant differ-ences in the levels of type Ⅲ procollagen peptide,laminin and type Ⅳ collagen between the mild to moderate group and the severe group(P＜0.05).Compared with the mild to moderate group,the levels of serum SOCS3 and GDF-15 in the severe group were significantly increased(P＜0.05).The results of Pearson corre-lation analysis showed that serum SOCS3 and GDF-15 in patients with T2DM combined with NAFLD were positively correlated with type Ⅲ procollagen peptide,laminin,and type Ⅳ collagen(P＜0.05).Serum SOCS3,GDF-1 5,type Ⅲ procollagen peptide,laminin and type Ⅳ collagen are risk factors affecting the degree of liver fibrosis in patients with T2DM combined with NAFLD(P＜0.05).The results of the ROC curve showed that the combined diagnosis of serum SOCS3 and GDF-15 for the degree of liver fibrosis in patients with T2DM complicated with NAFLD had the highest area under the curve(AUC),which was superior to the individual diagnosis of each(both P＜0.05),with a corresponding sensitivity of 69.08％and a specificity of 85.71％.The combined diagnosis of the degree of liver fibrosis in patients with T2DM complicated with NAFLD by serum SOCS3,GDF-15,type Ⅲ procollagen peptide,laminin,and type Ⅳ collagen had the highest AUC,which was superior to the individual diagnosis of each index(all P＜0.001),with a corresponding sensi-tivity of 89.47％and a specificity of 97.02％.Conclusion The levels of serum SOCS3 and GDF-15 are elevated in patients with T2DM combined with NAFLD,.The combined diagnosis of serum SOCS3,GDF-15,type Ⅲ procolla-gen peptide,laminin,and type Ⅳ collagen has a high value in the degree of liver fibrosis in patients.