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.

Animals ; Glycyrrhiza/adverse effects* ; Plant Extracts/toxicity* ; Rats ; Rats, Sprague-Dawley ; Sargassum

Objective: To investigate the effects of embryonic inflammation on the hippocampal synaptosomal-associated protein 25(SNAP-25) level and cognitive function in middle-aged. Methods: During gestational days 15-17, the CD-1 maternal mice received a daily intraperitoneal injection of lipopolysaccharides(LPS, 50 μg/kg) or the equal volume of normal saline, and the corresponding offspring were regarded as LPS group and CON group respectively. At the age of young(3-month-old) and middle-aged(15-month-old), the spatial learning and memory ability was assessed using Morris water maze(MWM), and the expression of hippocampal SNAP-25 protein was detected by immunohistochemical method and Western blot. Results: Compared with the 3-month CON group, the 15-month CON group had longer swimming distance(P<0.01), lower swimming distance percentage(P<0.01) in the target quadrant, and higher hippocampal subregions(CA1, CA3, DG) SNAP-25 levels(P<0.01). The same results were obtained in 15-month LPS group compared with 15-month CON group in learning and memory phase(P<0.05), and higher hippocampal subregions(CA1, DG) SNAP-25 levels(P<0.01). Pearson correlation analysis indicated that the hippocampal CA1 and CA3 subregions SNAP-25 level was positively correlated with the swimming distance, but negatively correlated with the percentage of swimming distance in the target quadrant. Conclusion Embryonic inflammation can accelerate the impairment of spatial learning and memory and the increase of hippocampal CA1 and CA3 subregions SNAP-25 protein in middle-aged CD-1 mice, and there may be a correlation between them.

Objective: To explore whether long-term potentiation(LTP) induced by brain-derived neurotrophic factor(BDNF) of hippocampus is involved in the process of maternal separation(MS) leading to impaired cognitive function of offspring in adolescence. Methods: The newborn CD-1 mice were randomly divided into maternal separation group(MS group) and control group(CON group). Mice in MS group were separated from the mother mice for 3 h every day from postnatal day 4 to 21 while no intervention was taken in the CON group. The spatial learning and memory ability was assessed using Morris water maze at the age of 3 months. Western blot and real-time quantitative PCR were used to detect the levels of BDNF and BDNF mRNA in the hippocampus. LTP of the hippocampal CA3-CA1 neural pathway was recorded using electrophysiological techniques. Results: Compared with CON group, the latency and distance of Morris water maze in maternal separation group were significantly longer(P<0.01). The percentage of time and distance in target quadrant during the memory phase in MS group were obviously lower than those in control group(P<0.05). The results of WB and Real-time quantitative PCR in MS group showed that the levels of BDNF and BDNF mRNA in MS group apparently decreased(P<0.05). Compared with CON group, MS group showed a significantly lower LTP in CA3-CA1 neural pathway(P<0.01). Conclusion The certain intensity of maternal separation can impair learning and memory function in young CD-1 male mice, which may be associated with decreased expression of BDNF and impaired LTP in the hippocampus.

Lianhuaqingwen (LHQW) capsule, a herb medicine product, has been clinically proved to be effective in coronavirus disease 2019 (COVID-19) pneumonia treatment. However, human exposure to LHQW components and their pharmacological effects remain largely unknown. Hence, this study aimed to determine human exposure to LHQW components and their anti-COVID-19 pharmacological activities. Analysis of LHQW component profiles in human plasma and urine after repeated therapeutic dosing was conducted using a combination of HRMS and an untargeted data-mining approach, leading to detection of 132 LHQW prototype and metabolite components, which were absorbed

The epidemic of coronavirus disease 2019 (COVID-19) puts higher demands on critical care medicine. Lots of studies have been conducted to solve COVID-19-related problems. Therefore, we reviewed the annual progress for COVID-19-related issues including antivirals threapies, respiratory support and immunomodulatory therapies and other critical issues, including the effect of antibiotic on mitochondrial damage and its relationship with sepsis, the goal and direction of antimicrobial de-escalation, drug prophylaxis of constipation, bleeding in gastrointestinal disorders and management of critical illness in the informalization era and so on. We hope to provide reference for clinical and scientific research work of the intensivists.

Objective To evaluate the effect of honokiol on the activation of transient receptor potential (TRP) channel in rat spinal dorsal root ganglion cells and pruritus in mouse models.Methods Healthy male ICR mice aged 4-6 weeks were used to establish histamine-induced and acetone/ether/water (AEW)-induced itching models separately.Totally,mice were randomly divided into 7 groups (histamineinduced model experiment) or 6 groups (AEW-induced model experiment):normal control group and model group both gavaged with sodium chloride physiological solution,solvent group gavaged with sodium carboxymethylcellulose solution,chlorphenamine group (only set up in the histamine-induced model experiment) gavaged with chlorphenamine,50,25 and 12.5 mg/kg honokiol groups gavaged with 50,25 and 12.5 mg/kg honokiol respectively.In the histamine-induced model experiment,the mice were all injected with histamine except the normal control group injected with sodium chloride physiological solution 24 hours after the gavage treatment,while the mice in the AEW-induced model experiment were all topically treated with AEW except the normal control group topically treated with sodium chloride physiological solution for 4 days,followed by gavage with different drugs.The anti-itch effect of each treatment was evaluated by counting the scratching frequency within 30 minutes.Rat spinal dorsal root ganglion (DRG)cells were isolated and subjected to a primary culture.Then,the DRG cells were divided into 6 groups:capsaicin or allyl isothiocyanate (AITC)-induced model group pre-incubated with Hank's balanced salt solution (HBSS),500 μmol/L capsazepine or 10 μmol/L HSC030031 group pre-incubated with capsazepine or HSC030031,solvent group pre-incubated with dimethyl sulfoxide (DMSO),3 honokiol groups preincubated with 7.81,15.63 and 31.25 mg/L honokiol respectively,and Ca2+ fluorescence imaging system was used to observe changes of Ca2+ influx in these cells after capsaicin or AITC stimulation.Statistical analysis was carried out with SPSS 20.0 software by using one-way analysis of variance and Dunnett-t test.Results In the histamine-induced mouse models,the scratching frequency was significantly lower in the 50 and 25 mg/kg honokiol groups than in the model group (21.88 and 21.14 vs.63.70,t =3.48,3.49 respectively,both P =0.003),while no significant difference in the scratching frequency was observed between the 12.5 mg/kg honokiol group and the model group (t =2.01,P =0.062).After the treatment with 50 mg/kg honokiol in the AEW-induced mouse models,the scratching frequency significantly decreased compared with the model group (61.4 vs.101.17,t =0.45,P =0.009),while there were no significant differences among the 25,12.5 mg/kg honokiol groups and the model group (all P ＞ 0.05).Compared with the capsaicin or AITC-induced model group,the increase of Ca2+ fluorescence signal in the DRG cells was significantly inhibited in the 31.25 mg/L honokiol group:at the 45th second,the rate of relative fluorescence intensity change (AF/F0) was 1.11 in the model group,but-0.11 in the 31.25 mg/L honokiol group in the capsaicin-induced model experiment,and 0.56 in the model group,but 0.00 in the 31.25 mg/L honokiol group in the AITC-induced model experiment.Conclusion Honokiol shows an inhibitory effect on mouse models of pruritus induced by histaminergic or non-histaminergic factors,likely by inhibiting Ca2+ influx through activated TRPV 1 and TRPA 1 channels in the DRG cells.

Objective To review the phenylalanine(Phe) and thyrotropin(TSH) results of national external quality assessment(EQA)in neonatal screening laboratory for 7 years,evaluate qualitity control level and summarize the problems in the screening measurement.Methods The Phe and TSH values in dried blood spot specimens on filter paper distributed by national center for clinical laboratory were measured by fluorometrie method and time-resolved fluorescence immunoassay respectively.The Phe and TSH EQA results from 2010 to 2016 year were analyzed.Results Five specimens were distributed by EQA organizer each time,and 3 times each year.(1) In the 105 specimens of 21 batches,the bias ranges of Phe and TSH measurement were wide and their average values were 2.21% and 0.98% respectively.(2) Quantitative results analysis :Six quantitative results of Phe measurement and two quantitative results of TSH measurement were out of aunty control(bias ≥30% or ≤-30%),accounting for 5.71%(6/105)and 1.9%(2/105) respectively.(3)Qualitative results analysis:The Phe and TSH qualitative analysis of specimens conformed to the expected results(100%).The result of every batch was up to 80%.Conclusion It is useful to evaluate the measurement competence,find out the problems in the Phe and TSH measurement in neonatal screening laboratory and resolve them in time.Consequently,it is useful to improve the quality of measurement,reduce the error and ensure the accuracy of results.

ObjectiveTo explore the role of peripheral blood intermediate monocytes in the pathogenesis of preeclampsia.MethodsFifty-two patients with established preeclampsia in Binhai County People's Hospital from October 2014 to October 2015, 42 healthy pregnant women and 42 healthy non-pregnant women were enrolled in this study. The percentage of intermediate monocyte subsets, ratio of positive cells and mean fluorescence intensity (MFI) of Toll-like receptor (TLR) 2, TLR4, CD64, and triggering receptor expressed on myeloid cell-1(TREM-1), and MFI of intracellular tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 were evaluated by flow cytometry. The concentrations of IL-6, IL-8, IL-1β, IL-12P70 and TNF-α in serum were analyzed using Luminex liquid phase chip technology. Independent two samplest-test, analysis of variance, Mann-WhitneyU test, Kruskal-Wallis test and Pearson correlation analysis were used for statistical analysis.ResultsPercentage of intermediate monocytes was higher in preeclampsia patients [10.4%(5.3%-19.9%)]than in healthy pregnant women [6.6%(4.9%-7.8%)], and both were higher than in non-pregnant women [3.8%(2.4%-5.0)%](allP<0.05). The ratio of TLR4 and CD64 positive intermediate monocytes [(60.1±12.5)%vs (24.9±8.8)%; (85.3±5.4)% vs (67.4±7.5)%](t were 15.416 and 13.437, bothP<0.05), and MFI of TLR4 (50.3±10.2 vs 26.8±8.6), TREM-1(35.6±4.1 vs 28.6±4.7) and CD64 (39.8±5.2 vs 28.9±4.8) (t were 11.898, 7.707 and 10.454, allP<0.05) were higher in preeclampsia patients than in healthy pregnant women. MFI of intracellular IL-6 (32.3±4.7 vs 28.6±3.5) and TNF-α (44.6±6.3 vs 36.7±8.3) in intermediate monocytes of preeclampsia patients was also significantly higher than that of healthy pregnant women (t were 4.239 and 5.245, bothP<0.05). Serum concentrations of IL-6, IL-8 and TNF-α were higher in preeclamptic patients than in healthy pregnant women and non-pregnant women (allP<0.05). Furthermore, a positive correlation was found between the percentage of intermediate monocytes and the serum levels of IL-6 and TNF-α in preeclamptic patients (r were 0.397 and 0.347, bothP<0.05).ConclusionsMonocyte subpopulations from preeclamptic patients are abnormally skewed toward intermediate monocytes which have high expressions of TLR4, TREM-1 and CD64, and secret more proinflammatory cytokines such as IL-6 and TNF-α. Therefore, intermediate monocytes are specifically altered in preeclamptic patients and may play a role in the pathophysiology of preeclampsia.

Alpha-ketoglutarate (AKG) is a key molecule in the Krebs cycle determining the overall rate of the citric acid cycle of the organism. It is a nitrogen scavenger and a source of glutamate and glutamine that stimulates protein synthesis and inhibits protein degradation in muscles. AKG as a precursor of glutamate and glutamine is a central metabolic fuel for cells of the gastrointestinal tract as well. AKG can decrease protein catabolism and increase protein synthesis to enhance bone tissue formation in the skeletal muscles and can be used in clinical applications. In addition to these health benefits, a recent study has shown that AKG can extend the lifespan of adult Caenorhabditis elegans by inhibiting ATP synthase and TOR. AKG not only extends lifespan, but also delays age-related disease. In this review, we will summarize the advances in AKG research field, in the content of its physiological functions and applications.
Adenosine Triphosphate ; Adult ; Bone and Bones ; Caenorhabditis elegans ; Citric Acid Cycle ; Gastrointestinal Tract ; Glutamic Acid ; Glutamine ; Humans ; Insurance Benefits ; Metabolism ; Muscle, Skeletal ; Muscles ; Nitrogen ; Proteolysis