OBJECTIVE To establish a method for simultaneously determining three anti-tuberculosis drugs in the plasma of patients with spinal tuberculosis and apply it in clinical practice. METHODS LC-MS/MS method was established for the quantitative determination of the concentrations of isoniazid， rifampicin and pyrazinamide in the plasma of patients with spinal tuberculosis， using diphenhydramine as the internal standard. The determination was carried out using Chemalink CM-C18T column， with mobile phase consisting of 0.1% formic acid-methanol solution （gradient elution）， at the flow rate of 0.4 mL/min and an injection volume of 2 μL. Multiple reaction monitoring was conducted using an electrospray ionization source in positive ion mode. The ion pairs used for quantitative analysis were m/z 138.0→121.0（ for isoniazid）， m/z 823.3→791.3（ for rifampicin）， m/z 124.1→ 79.0 （for pyrazinamide）， and m/z 256.0→167.0 （for diphenhydramine）. Fifty-three patients diagnosed with spinal tuberculosis in Qinghai Provincial People’s Hospital from January 2023 to June 2025 were selected， and the plasma concentrations of isoniazid， rifampicin and pyrazinamide in these patients were measured using the above method. RESULTS The linear ranges for isoniazid， rifampicin and pyrazinamide were 0.5-16， 2-64， and 2.5-80 μg/mL， respectively （r≥0.998 7）. The accuracy ranged from 90.20% to 108.64% （n＝5）. RSDs for intra-day precision were all less than 6.63% （n＝5）， while those for inter-day precision were all less than 8.42% （n＝3）. The matrix effects ranged from 88.60% to 115.41% （n＝5）. The relative deviations in the stability tests were all within the ±15% range， and the carry-over effect did not interfere with the determination. The results of clinical application showed that the mean plasma drug concentrations of isoniazid， rifampicin and pyrazinamide in patients with spinal tuberculosis were （3.62±2.80）， （8.55±4.57）， and （20.12±6.56） μg/mL， respectively. The incidences of plasma drug concentrations falling below the effective peak concentrations were 49.06%， 58.49% and 60.38%， respectively. CONCLUSIONS The method established in this study is rapid， accurate， and demonstrates good stability， making it suitable for clinical monitoring of the plasma concentrations of isoniazid， rifampicin and pyrazinamide in patients with spinal tuberculosis.

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

Objective To explore the effect of Rhizoma Polygonati(RP)on arterial aging in naturally aging Wistar rats.Methods SPF Wistar rats aged 72 weeks were divided randomly divided into 4 groups:an old group and RP low,medium,and high-dose groups(n=14 rats per group).Another 14 male SPF Wistar rats aged 8～12 weeks were selected as the young group.Rats in the RP high,medium,and low-dose groups were administered with 4,2,and 1 g/kg RP,respectively,by gavage,and rats in the old and young groups were given the same amount of distilled water once a day for 12 weeks.Seven rats from each group were sacrificed under anesthesia at weeks 4 and 12 and aortas were isolated.The relative smooth muscle cell(SMC)and collagen fiber(CF)contents were analyzed,total antioxidant capacity(T-AOC),glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),and malondialdehyde(MDA)levels were measured,and the expression levels of cell cycle-associated proteins in arterial tissue were detected by Western Blot.Results Rats in the old group showed obvious signs of vascular aging but there was no significant changes in arterial vascular tissue indexes in the old group with increased age.Aortas were obviously injured,relative contents of SMC and CF were significantly increased(P＜0.01),T-AOC,SOD,and GSH-Px contents were significantly decreased and MDA was increased(P＜0.01)in the old group compared with the young group at 4 and 8 weeks,and expression levels of cell cycle-associated proteins were significantly up-regulated(P＜0.01).RP intervention significantly decreased the relative SMC and CF contents and MDA levels(P＜0.05 or P＜0.01)and significantly increased T-AOC,SOD,and GSH-Px(P＜0.05 or P＜0.01).Expression levels of cell cycle-associated proteins were also significantly decreased(P＜0.05 or P＜0.01).High-dose RP had the greatest effect.Conclusions Arterial aging is relatively stable in the short term in naturally aging rats.RP could delay arterial aging in naturally aging rats by regulating the level of oxidative stress and the expression of cell cycle-associated proteins.

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 search, evaluate, and summarize the best evidence for postpartum contraceptive health guidance, providing evidence-based support for clinical healthcare providers in implementing standardized contraceptive counseling and management.Methods:A systematic search was conducted across guideline repositories, professional association websites, and databases for literature related to postpartum contraception guidance, including guidelines, best practices, expert consensus, and systematic reviews, with a search timeframe from database inception to December 2023. Four researchers independently evaluated the quality of the included studies, extracted relevant data, and synthesized evidence from eligible literature.Results:According to the inclusion and exclusion criteria, 18 documents were included, comprising 5 guidelines, 2 clinical decision-making documents, 1 best practice document, 4 expert consensus statements, and 6 meta-analyses or systematic reviews. Totally 46 pieces of best evidence were summarized from 9 aspects, including health educators, health education recipients, assessment, planning, mode and content of health education, available contraceptive methods, evaluation index of health education, and considerations.Conclusion:This study systematically synthesizes the best available evidence on postpartum contraceptive health guidance. It emphasizes strengthening the competencies of clinical practitioners, supported by structured assessments and standardized guidance, to improve the feasibility and accessibility of contraceptive services. It further highlights the importance of ensuring the long-term sustainability of contraceptive plans and integrating digital tools to enhance the precision and coverage of guidance, ultimately reducing unintended and short-interval pregnancies and safeguarding women's reproductive health.

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 predictive value of albumin-bilirubin score(ALBI),tumor burden score(TBS)and lac-tate dehydrogenase(LDH)level in predicting the prognosis of HCC patients after radical hepatectomy.Methods The clinical data of pa-tients who underwent radical hepatectomy for primary liver cancer in the Department of General Surgery,The Second Hospital of Lanzhou University from January 2011 to January 2022 were retrospectively collected,including the patients'basic personal information,relevant laboratory tests,imaging data,pathological reports and follow-up data.This study involved 162 patients.The patients were divided into survival group and death group according to their survival status at the last follow-up,and the area under the receiver operating character-istic curve(AUC)was used to estimate the prediction accuracy of different models.Results During the study period,a total of 79 pa-tients(48.7％)died.The median survival time was 46.5months.AFP and LDH were identified as two independent risk factors for over-all survival after surgery by univariate and multivariate analysis(P＜0.05).The AUC of ALBI-TBS-LDH score for predicting death af-ter radical hepatectomy was higher than that of ALBI score and TBS score.According to the optimal Youden index,when the optimal diag-nostic cutoff value of ALBI-TBS-LDH score was-0.18,the sensitivity and specificity of predicting death were 63.4％and 71.8％,respectively.The 1-year,3-year and 5-year overall survival rates of patients with ALBI-TBS-LDH score＜-0.18 were 95.1％,80.5％and 39.0％,respectively;the 1-year,3-year and 5-year overall survival rates of patients with score≥-0.18 were 86.6％,59.7％and 22.4％,respectively,with statistically significant differences(P＜0.001).Conclusion The ALBI-TBS-LDH score can be used as a simple tool to evaluate the prognosis of HCC patients after liver resection,but its role still needs further research and verifica-tion.Patients with high ALBI-TBS-LDH scores have a higher incidence of postoperative death and a shorter survival time than those with low scores.

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 investigate the clinical manifestations and prognosis of patients with anti-leucine rich glioma inactivated 1（LGI1）antibody encephalitis. Methods A retrospective analysis was performed for the data of patients with anti-LGI1 antibody encephalitis who were hospitalized in Department of Neurology，Xuanwu Hospital，Capital Medical University，from September 2019 to December 2023，including clinical features，treatment，and prognosis. Results A total of 80 patients were enrolled，including 48 male patients and 32 female patients. Of all 80 patients，51（63.8%）had the initial symptom of epilepsy，13（16.3%）had the initial symptom of faciobrachial dystonic seizures，26（32.5%）had the initial symptom of cognitive impairment，and 12（15%）had the initial symptom of mental and behavioral disorders. As for comorbidities，there were 28 patients（35%）with hyponatremia，22（27.5%）with Hashimoto's thyroiditis，and 5（6.25%）with tumor. All 80 patients tested positive for serum LGI1 antibody，and 69（86.3%）tested positive for LGI1 antibody in cerebrospinal fluid. Among the 80 patients，37（46.25%）had abnormalities on magnetic resonance imaging，and 30 had abnormalities on 18F-FDG PET/CT. There were 50 patients with abnormalities on video electroencephalography. A total of 77 patients received first-line immunotherapy，and after follow-up for 1 year，19.4% of the patients had the sequela of seizure，59.7% of the patients had deterioration of the memory，and 48.6% of the patients had mental and behavioral disorders. Conclusion Epilepsy，cognitive impairment，and mental and behavior disorders are the most common manifestations of anti-LGI1 antibody encephalitis. There is generally a good prognosis after immunotherapy，with the sequelae of cognitive impairment and mental and behavioral disorders.

Objective To explore the effect of Rhizoma Polygonati(RP)on arterial aging in naturally aging Wistar rats.Methods SPF Wistar rats aged 72 weeks were divided randomly divided into 4 groups:an old group and RP low,medium,and high-dose groups(n=14 rats per group).Another 14 male SPF Wistar rats aged 8～12 weeks were selected as the young group.Rats in the RP high,medium,and low-dose groups were administered with 4,2,and 1 g/kg RP,respectively,by gavage,and rats in the old and young groups were given the same amount of distilled water once a day for 12 weeks.Seven rats from each group were sacrificed under anesthesia at weeks 4 and 12 and aortas were isolated.The relative smooth muscle cell(SMC)and collagen fiber(CF)contents were analyzed,total antioxidant capacity(T-AOC),glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),and malondialdehyde(MDA)levels were measured,and the expression levels of cell cycle-associated proteins in arterial tissue were detected by Western Blot.Results Rats in the old group showed obvious signs of vascular aging but there was no significant changes in arterial vascular tissue indexes in the old group with increased age.Aortas were obviously injured,relative contents of SMC and CF were significantly increased(P＜0.01),T-AOC,SOD,and GSH-Px contents were significantly decreased and MDA was increased(P＜0.01)in the old group compared with the young group at 4 and 8 weeks,and expression levels of cell cycle-associated proteins were significantly up-regulated(P＜0.01).RP intervention significantly decreased the relative SMC and CF contents and MDA levels(P＜0.05 or P＜0.01)and significantly increased T-AOC,SOD,and GSH-Px(P＜0.05 or P＜0.01).Expression levels of cell cycle-associated proteins were also significantly decreased(P＜0.05 or P＜0.01).High-dose RP had the greatest effect.Conclusions Arterial aging is relatively stable in the short term in naturally aging rats.RP could delay arterial aging in naturally aging rats by regulating the level of oxidative stress and the expression of cell cycle-associated proteins.