OBJECTIVES: Tiletamine, a veterinary anesthetic, has emerged as a novel psychoactive substance and has been abused in many parts of the world, causing great harm to public health. However, the sensitivity of existing detection methods cannot meet the needs of forensic practice. This study aims to establish an ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of tiletamine and its metabolite desethyltiletamine in human biological samples, and to verify its applicability in forensic practice. METHODS: SKF_525A was used as the internal standard. Biological samples were extracted with acetonitrile containing 1 ng/mL SKF_525A, vortexed for 10 min, ultrasonicated for 20 min, centrifuged at 10 000 r/min for 10 min, and 500 μL of the supernatant was filtered through a 0.22 μm membrane. Analyses were performed using an ACQUITY UPLC H-Class PLUS system and an XEVO TQ-S Micro triple quadrupole mass spectrometer. An ACQUITY UPLC^® BEH C18 (1.7 µm, 2.1 mm×100 mm) column at a flow rate of 0.3 mL/min was used, and four mobile phase systems were tested to optimize separation. Detection used positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode, with quantifier ion transitions of mass to charge 224.043→179.016 for tiletamine and mass to charge 196.08→151.06 for desethyltiletamine. Calibration curves were established over 0.1-200 ng/mL in spiked blood samples. The linear range, limit of detection (LOD), and limit of quantification (LOQ) were determined. Low (5 ng/mL), medium (20 ng/mL), and high (100 ng/mL) concentrations of tiletamine were spiked into blood, liver, and kidney to evaluate precision, accuracy, matrix effect, recovery, and stability. Finally, actual forensic case samples were tested to validate applicability. RESULTS: The established UPLC-MS/MS method achieved simultaneous detection of tiletamine and desethyltiletamine in human biological samples, with retention times of 3.42 min and 2.82 min, respectively. Using mobile phase A (20 mmol/L ammonium acetate and 0.1% formic acid in water) and mobile phase B (acetonitrile) produced the best separation. In blood, tiletamine showed good linearity from 0.1-200 ng/mL (r=0.992, R²=0.983), LOD 0.03 ng/mL, LOQ 0.1 ng/mL, recovery 92%-107%, and matrix effect 71%-99%. In liver and kidney, recoveries were 91%-98% and 93%-104%, and matrix effects were 69%-96% and 72%-100%, respectively. Intra- and inter-day precision [expressed as relative standard deviation (RSD)] and accuracy [expressed as relative error (RE)] were within 15%, and samples were stable at -20 ℃. Tiletamine was detected in actual case samples at 0.37 μg/mL (blood), 0.15 μg/g (liver), 0.11 μg/g (kidney) in case 1, and 8.75 ng/mL (blood) in case 2; desethyltiletamine was also detected in blood. CONCLUSIONS The UPLC-MS/MS method is efficient, accurate, and sensitive, and is suitable for detecting tiletamine and desethyltiletamine in human biological samples.
Tandem Mass Spectrometry/methods* ; Humans ; Chromatography, High Pressure Liquid/methods* ; Substance Abuse Detection/methods* ; Liquid Chromatography-Mass Spectrometry

One of the basic questions in the aging field is whether there is a fundamental difference between the aging of lower invertebrates and mammals. A major difference between the lower invertebrates and mammals is the abundancy of noncoding RNAs, most of which are not conserved. We have previously identified a noncoding RNA Terc-53 that is derived from the RNA component of telomerase Terc. To study its physiological functions, we generated two transgenic mouse models overexpressing the RNA in wild-type and early-aging Terc-/- backgrounds. Terc-53 mice showed age-related cognition decline and shortened life span, even though no developmental defects or physiological abnormality at an early age was observed, indicating its involvement in normal aging of mammals. Subsequent mechanistic study identified hyaluronan-mediated motility receptor (Hmmr) as the main effector of Terc-53. Terc-53 mediates the degradation of Hmmr, leading to an increase of inflammation in the affected tissues, accelerating organismal aging. adeno-associated virus delivered supplementation of Hmmr in the hippocampus reversed the cognition decline in Terc-53 transgenic mice. Neither Terc-53 nor Hmmr has homologs in C. elegans. Neither do arthropods express hyaluronan. These findings demonstrate the complexity of aging in mammals and open new paths for exploring noncoding RNA and Hmmr as means of treating age-related physical debilities and improving healthspan.
Animals ; Mice ; RNA, Untranslated/metabolism* ; Aging/genetics* ; Mice, Transgenic ; Telomerase/metabolism* ; RNA/genetics* ; Hippocampus/metabolism* ; Humans ; Mice, Inbred C57BL

Objective To investigate the clinical treatment effect and safety of 3D microscopy technology for adjuvant neuraxial tumor resection on neuraxial tumor diseases.Methods A total of 37 patients with neuraxial tumors treated from January 2019 to July 2023,15 patients treated with 3D microscope tumor resection(3D group),and 22 patients treated with general microscope tumor resection(ordinary group)were analyzed.The perioperative indexes,clinical efficacy indexes and safety indexes were compared between the two groups.Results The operation time was(223.78±46.46)min in the ordinary group and(182.93±39.28)min in the 3D group,which was significantly lower than that in the ordinary group(P<0.05),and there was no significant difference in other perioperative indicators between the two groups(P>0.05).All patients had significantly reduced their postoperative pain symptoms and recovered their neurological function to a certain extent.There were statistically significant differences between the two groups(P<0.01),but there was no statistical difference between the two groups(P>0.05),and the McCormick spinal cord function rating was grade I.at one year after surgery.Conclusion The use of 3D microscopy and general microscopy for neuraxial tumor surgery has good clinical efficacy.However,the operation time can be significantly shortened under 3D microscopy,thereby reducing the risk of surgical complications and has better clinical safety.

OBJECTIVE To understand the disinfectant-resistant genes in the gram-negative bacteria isolated from the dirt retention on air recure filters of air conditioners and observe the drug resistance.METHODS The dirt re-tention samples were collected from the air return filters of air conditioners of some wards in 3 hospitals of Wuhan(Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology,Wuhan Central Hospital,and Hubei Provincial Maternal and Child Health Hospital)from 2018 to 2024.The gram-negative bac-teria were screened out,the disinfectant-resistant genes in the strains were detected by polymerase chain reaction(PCR),and the results of drug susceptibility test were analyzed.RESULTS Of 354 dirt retention samples that were collected from the air return filers of air conditioners,77 were detected with 138 strains of gram-negative bacteria,87 of which were Acinetobacter baumannii,50 were Enterobacteriaceae,and 1 was Pseudomonas aerug-inosa.The detection rates of qacEΔ1,qacEΔ1-sul1,aceI and qacA/B were 73.19％,82.61％,69.57％and 2.90％,respectively.None of the strains were detected with qacC,qacH or qacJ.The result of drug susceptibility test showed that 76.81％of the gram-negative bacteria were resistant to at least 1 type of antibiotic;93 strains of multidrug-resistant gram-negative bacteria were isolated,most of which were isolated from intensive care unit(ICU).The detection rates of qacEΔ1 and qacEΔ1-sul1 were higher in the drug-resistant strains than those in the non-drug-resistant strains;there were significant differences in the drug resistance rates to carbapenems,quin-olones and β-lactams between the qacEΔ1-sul 1-positive strains and the qacEΔ1-sul1-negative strains(P＜0.05).CONCLUSIONS There are drug-resistant gram-negative bacteria contaminations in some wards of the 3 hospitals in Wuhan.The carrying rates of disinfectant-resistant genes of the strains are high,and the strains show varying degree of resistance to the commonly used antibiotics;the strains carrying the qacEΔ1-sul1 have certain statistical association with the drug resistance.It is suggested that the hospital should take targeted disinfec-tion measures for the environment and reasonably use antibiotics.

OBJECTIVE To understand the disinfectant-resistant genes in the gram-negative bacteria isolated from the dirt retention on air recure filters of air conditioners and observe the drug resistance.METHODS The dirt re-tention samples were collected from the air return filters of air conditioners of some wards in 3 hospitals of Wuhan(Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology,Wuhan Central Hospital,and Hubei Provincial Maternal and Child Health Hospital)from 2018 to 2024.The gram-negative bac-teria were screened out,the disinfectant-resistant genes in the strains were detected by polymerase chain reaction(PCR),and the results of drug susceptibility test were analyzed.RESULTS Of 354 dirt retention samples that were collected from the air return filers of air conditioners,77 were detected with 138 strains of gram-negative bacteria,87 of which were Acinetobacter baumannii,50 were Enterobacteriaceae,and 1 was Pseudomonas aerug-inosa.The detection rates of qacEΔ1,qacEΔ1-sul1,aceI and qacA/B were 73.19％,82.61％,69.57％and 2.90％,respectively.None of the strains were detected with qacC,qacH or qacJ.The result of drug susceptibility test showed that 76.81％of the gram-negative bacteria were resistant to at least 1 type of antibiotic;93 strains of multidrug-resistant gram-negative bacteria were isolated,most of which were isolated from intensive care unit(ICU).The detection rates of qacEΔ1 and qacEΔ1-sul1 were higher in the drug-resistant strains than those in the non-drug-resistant strains;there were significant differences in the drug resistance rates to carbapenems,quin-olones and β-lactams between the qacEΔ1-sul 1-positive strains and the qacEΔ1-sul1-negative strains(P＜0.05).CONCLUSIONS There are drug-resistant gram-negative bacteria contaminations in some wards of the 3 hospitals in Wuhan.The carrying rates of disinfectant-resistant genes of the strains are high,and the strains show varying degree of resistance to the commonly used antibiotics;the strains carrying the qacEΔ1-sul1 have certain statistical association with the drug resistance.It is suggested that the hospital should take targeted disinfec-tion measures for the environment and reasonably use antibiotics.

Objective To investigate the clinical treatment effect and safety of 3D microscopy technology for adjuvant neuraxial tumor resection on neuraxial tumor diseases.Methods A total of 37 patients with neuraxial tumors treated from January 2019 to July 2023,15 patients treated with 3D microscope tumor resection(3D group),and 22 patients treated with general microscope tumor resection(ordinary group)were analyzed.The perioperative indexes,clinical efficacy indexes and safety indexes were compared between the two groups.Results The operation time was(223.78±46.46)min in the ordinary group and(182.93±39.28)min in the 3D group,which was significantly lower than that in the ordinary group(P<0.05),and there was no significant difference in other perioperative indicators between the two groups(P>0.05).All patients had significantly reduced their postoperative pain symptoms and recovered their neurological function to a certain extent.There were statistically significant differences between the two groups(P<0.01),but there was no statistical difference between the two groups(P>0.05),and the McCormick spinal cord function rating was grade I.at one year after surgery.Conclusion The use of 3D microscopy and general microscopy for neuraxial tumor surgery has good clinical efficacy.However,the operation time can be significantly shortened under 3D microscopy,thereby reducing the risk of surgical complications and has better clinical safety.

ObjectiveTo explore the possible mechanism of Osteoking （OK） on postmenopausal osteoporosis （PMOP）. MethodForty adult female mice were randomly divided into a sham operation （Sham） group， osteoporosis model （OVX） group， estradiol intervention （E₂） group， and OK group， with 10 mice in each group. The modeling was completed by conventional back double incision ovariectomy， and the corresponding drugs were given one week later. After 12 weeks， the body mass and uterine index of mice were measured， and the pathological changes of bone tissue and the number of osteoclasts （OCs） were determined by hematoxylin-eosin （HE） and tartrate-resistant acid phosphatase （TRAP） staining， respectively. Bone mineral density （BMD）， trabecular number （Tb.N）， trabecular separation （Tb.Sp）， and bone volume fraction （BV/TV） were measured by microcomputed tomography （Micro-CT）. The maximum load of the femur was detected by a three-point bending test. The contents of tumor necrosis factor-α （TNF-α） and bone resorption marker C-terminal telopeptide of type Ⅰ collagen （CTX-1） were measured by enzyme linked immunosorbent assay （ELISA）. The protein expression levels of nuclear factor-kappa B p65 （NF-κB p65）， phosphorylated nuclear factor-kappa B p65 （p-NF-κB p65）， nuclear factor kappa B inhibitor alpha （IκBα）， phosphorylated nuclear factor kappa B alpha （p-IκBα）， nuclear factor of activated T cells 1 （NFATc1）， and proto-oncogene （c-Fos） were detected by Western blot. The mRNA expressions of OCs-related specific genes matrix metalloproteinase-9 （MMP-9）， NFATc1， TRAP， cathepsin K （CTSK）， and c-Fos were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the Sham group， the uterine index decreased significantly in the OVX group， and the body mass （BMI） increased significantly. The structure of bone trabeculae was completely damaged， and the number of OCs increased. BMD， Tb.N， BV/TV， and maximum load decreased， while Tb.Sp was up-regulated. The levels of TNF-α and CTX-1 in serum were up-regulated. The protein expressions of c-Fos， p-NF-κB p65/NF-κB p65， NFATc1， and p-IκBα/IκBα were increased. The mRNA expressions of NFATc1， c-Fos， CTSK， TRAP， and MMP-9 were up-regulated （P<0.05， P<0.01）. Compared with the OVX group， the body mass of the OK and E₂ groups decreased， and the uterine index increased. The bone trabeculae increased， and the number of OCs decreased. BMD， Tb.N， BV/TV， and maximum load increased， while Tb.Sp decreased. The levels of TNF-α and CTX-1 in serum were decreased. The protein expressions of c-Fos， p-NF-κB p65/NF-κB p65， NFATc1， and p-IκBα/IκBα were decreased， and the mRNA expressions of NFATc1， c-Fos， CTSK， TRAP， and MMP-9 were decreased （P<0.05， P<0.01）. ConclusionOK can inhibit the NF-κB/NFATc1 signaling pathway and reduce bone mass loss by reducing the level of inflammatory injury factors in PMOP mice， which is one of the mechanisms for treating PMOP.

Sepsis is a multi-organ dysfunction syndrome caused by infection and immune dysfunction, with a high mortality rate. It affects multiple important organs such as the heart, lungs, kidneys, liver, and brain. Establishing corresponding animal models of organ dysfunction syndrome is an essential step in clarifying its pathogenesis, researching potential effective drugs, and evaluating the effectiveness and safety of treatment plans. This article first summarizes classic modeling methods for sepsis related organ injury, including the destruction of intestinal barrier tissue integrity and the implantation of pathogens or toxic drugs. The former mainly includes cecal ligation and puncture, ascending colon stent implantation, and cecal ligation incision. The latter is divided into intraperitoneal injection, intravenous injection, and intratracheal administration based on the clinical infection route being simulated. Cecal ligation and puncture and lipopolysaccharide intraperitoneal injection are the most commonly used methods. Secondly, this article summarizes the common modeling methods and evaluation methods for animal models of sepsis-induced cardiomyopathy, acute lung injury, acute kidney injury, acute liver injury, and brain dysfunction. It points out that almost all organ injuries use classic modeling methods, and different organ injury models have additional modifications according to their different pathogenesis. For example, in addition to the classic modeling methods, lipopolysaccharide instillation in the trachea is more effective in modeling acute lung injury as it better simulates lung barrier dysfunction. Cecal ligation and puncture followed by Pseudomonas instillation in the trachea in a secondary challenge model better represents sepsis-induced acute kidney injury. Intraperitoneal injection of galactosamine is a mature modeling method of sepsis-induced acute liver injury. Intracerebral injection of lipopolysaccharide is a feasible model of sepsis-associated encephalopathy. In addition to the different modeling methods, there are differences in the administration time, dosage and experimental time points according to the different experimental purposes. This article reviews the research progress of animal experimental models for sepsis-induced cardiomyopathy, acute lung injury, acute kidney injury, acute liver injury, and brain dysfunction, aiming to provide a reference for the selection of animal experimental models and optimization of experimental design.

Objective:To investigate the bidirectional causal relationship between schizophrenia and suicide or intentional self-harm using a Mendelian randomization (MR) approach.Methods:Using pooled data from a large-scale genome-wide association study, genetic loci that were independent of each other and associated with schizophrenia and suicide or intentional self-harm in populations of European ancestry were selected as instrumental variables. The data for schizophrenia included 76 755 patients with schizophrenia and 243 649 controls. Data for suicide or intentional self-harm included 1 058 patients with suicide or intentional self-harm and 307 942 controls. The inverse variance weighting (IVW) and weighted median methods of MR analysis were used to explore the bidirectional causality of schizophrenia and suicide or intentional self-harm using the ratio of ratios ( OR) as an evaluation index. Heterogeneity between individual single nucleotide polymorphism (SNP) was assessed by Cochran′s Q test. Mendelian randomization polymorphism residuals and outliers (MR-PRESSO) were used to detect SNP outliers, and MR-Egger regression tests were performed to examine the horizontal pleiotropy of SNPs. A "leave-one-out" sensitivity analysis was performed to test whether the MR study was influenced by individual SNPs. Results:The IVW approach showed that schizophrenia increased the risk of suicide or intentional self-harm ( OR=1.297, 95% CI=1.154-1.457, P<0.001). In addition, the above relationship was confirmed in the weighted median ( OR=1.232, 95% CI=1.038-1.461, P=0.017). Conversely, suicide or intentional self-harm was not causally associated with schizophrenia ( OR=0.969, 95% CI=0.927—1.012, P=0.157), again confirmed by the weighted median method. The Cochran′s Q test, MR-PRESSO and MR-Egger regression tests did not show heterogeneity or horizontal pleiotropy between instrumental variables. The "leave-one-out" sensitivity analysis confirmed that no single SNP had a disproportionate effect on the overall outcome. Conclusion:Schizophrenia is significantly and causally associated with an increased risk of suicide or intentional self-harm.

Objective:To investigate the bidirectional causal relationship between schizophrenia and suicide or intentional self-harm using a Mendelian randomization (MR) approach.Methods:Using pooled data from a large-scale genome-wide association study, genetic loci that were independent of each other and associated with schizophrenia and suicide or intentional self-harm in populations of European ancestry were selected as instrumental variables. The data for schizophrenia included 76 755 patients with schizophrenia and 243 649 controls. Data for suicide or intentional self-harm included 1 058 patients with suicide or intentional self-harm and 307 942 controls. The inverse variance weighting (IVW) and weighted median methods of MR analysis were used to explore the bidirectional causality of schizophrenia and suicide or intentional self-harm using the ratio of ratios ( OR) as an evaluation index. Heterogeneity between individual single nucleotide polymorphism (SNP) was assessed by Cochran′s Q test. Mendelian randomization polymorphism residuals and outliers (MR-PRESSO) were used to detect SNP outliers, and MR-Egger regression tests were performed to examine the horizontal pleiotropy of SNPs. A "leave-one-out" sensitivity analysis was performed to test whether the MR study was influenced by individual SNPs. Results:The IVW approach showed that schizophrenia increased the risk of suicide or intentional self-harm ( OR=1.297, 95% CI=1.154-1.457, P<0.001). In addition, the above relationship was confirmed in the weighted median ( OR=1.232, 95% CI=1.038-1.461, P=0.017). Conversely, suicide or intentional self-harm was not causally associated with schizophrenia ( OR=0.969, 95% CI=0.927—1.012, P=0.157), again confirmed by the weighted median method. The Cochran′s Q test, MR-PRESSO and MR-Egger regression tests did not show heterogeneity or horizontal pleiotropy between instrumental variables. The "leave-one-out" sensitivity analysis confirmed that no single SNP had a disproportionate effect on the overall outcome. Conclusion:Schizophrenia is significantly and causally associated with an increased risk of suicide or intentional self-harm.