Liquid chromatography-electrospray ionization tandem mass spectrometry(LC-ESI-MS)is a widely utilized technique for in vivo pharmaceutical analysis.Ionization interference within electrospray ion source,occurring between drugs and metabolites,can lead to signal variations,potentially compromising quantitative accuracy.Currently,method validation often overlooks this type of signal interference,which may result in systematic errors in quantitative results without matrix-matched calibration.In this study,we conducted an investigation using ten different groups of drugs and their corresponding me-tabolites across three LC-ESI-MS systems to assess the prevalence of signal interference.Such in-terferences can potentially cause or enhance nonlinearity in the calibration curves of drugs and metabolites,thereby altering the relationship between analyte response and concentration for quanti-fication.Finally,we established an evaluation scheme through a step-by-step dilution assay and employed three resolution methods:chromatographic separation,dilution,and stable labeled isotope internal standards correction.The above strategies were integrated into the method establishment process to improve quantitative accuracy.

Purpose: Prostate cancer (PCa) is an epithelial malignancy that originates in the prostate gland and is generally categorized into low, intermediate, and high-risk groups. The primary diagnostic indicator for PCa is the measurement of serum prostate-specific antigen (PSA) values. However, reliance on PSA levels can result in false positives, leading to unnecessary biopsies and an increased risk of invasive injuries. Therefore, it is imperative to develop an efficient and accurate method for PCa risk stratification. Many recent studies on PCa risk stratification based on clinical data have employed a binary classification, distinguishing between low to intermediate and high risk. In this paper, we propose a novel machine learning (ML) approach utilizing a stacking learning strategy for predicting the tripartite risk stratification of PCa. Methods: Clinical records, featuring attributes selected using the lasso method, were utilized with 5 ML classifiers. The outputs of these classifiers underwent transformation by various nonlinear transformers and were then concatenated with the lasso-selected features, resulting in a set of new features. A stacking learning strategy, integrating different ML classifiers, was developed based on these new features. Results: Our proposed approach demonstrated superior performance, achieving an accuracy of 0.83 and an area under the receiver operating characteristic curve value of 0.88 in a dataset comprising 197 PCa patients with 42 clinical characteristics. Conclusions This study aimed to improve clinicians’ ability to rapidly assess PCa risk stratification while reducing the burden on patients. This was achieved by using artificial intelligence-related technologies as an auxiliary method for diagnosing PCa.

Objective:To investigate the efficacy and safety of dual antiplatelet therapy (DAPT) in patients with minor ischemic stroke (MIS) and tiny unruptured intracranial aneurysm (UIA).Methods:Patients with MIS and tiny UIA admitted to the Department of Neurology, Weifang People's Hospital from October 1, 2022 to February 29, 2024 were included retrospectively. MIS was defined as baseline National Institutes of Health Stroke Scale (NIHSS) score ≤3. Tiny UIA was defined as UIA with a diameter of ≤3 mm. According to the antiplatelet therapy regimen, the patients were divided into an aspirin alone group and an aspirin+clopidogrel DAPT group. The main outcome measure was the clinical outcome at 90 days after onset. The modified Rankin Scale (mRS) score 0-1 was defined as a good outcome and >1 was defined as a poor outcome. Secondary outcome measures included aneurysm rupture, cerebral hemorrhage, and recurrence of cerebral ischemic events. Multivariate logistic regression analysis was used to identify the independent influencing factors for poor outcome. Results:A total of 183 patients with MIS and tiny UIA were included, including 108 males (59.0%), median aged 68 years (interquartile range, 61-73 years). All the UIAs were solitary. The mRS score of all patients before onset was 0; 152 patients (83.1%) had good outcome at 90 days after onset, 31 (16.9%) had poor outcome, and no UIA occurred rupture bleeding. Of the 94 patients (51.4%) who received aspirin monotherapy, 14 patients (14.9%) experienced recurrent cerebral ischemic events during follow-up, and 73 (77.7%) had good outcome. Of the 89 patients (48.6%) who received DAPT, 5 (5.6%) experienced recurrent ischemic events during follow-up, and 79 (88.8%) had good outcome. The recurrence rate of cerebral ischemic events in the aspirin group was significantly higher than that in the DAPT group ( χ2=4.227, P=0.040), while the good outcome rate was significantly lower than that in the DAPT group ( χ2=4.006, P=0.045). Multivariate logistic regression analysis showed that baseline NIHSS score was an independent risk factor for poor outcome (odds ratio 4.597, 95% confidence interval 1.864-11.339; P=0.001), while DAPT was an independent protective factor for good outcome (odds ratio 0.265, 95% confidence interval 0.079-0.892; P=0.032). Conclusion:Compared with aspirin monotherapy, the short-term combination of aspirin and clopidogrel in patients with MIS and tiny UIA may improve the outcome, reduces the recurrence of cerebral ischemic events, and has good safety.

Ectoine is an amino acid derivative and an important natural product in halophilic microorganisms. It plays an important role in protecting cells and stabilizing biological macromolecules, and can be widely used in biomedical fields such as drug preparation adjuvants, organ transplantation and preservation, skin wound repair and cosmetics. Due to the medical value and commercial market demand of ectoine, this article summarized the recent advances in the microbial production of ectoine, including the mutation and breeding of hyper-producing strains, construction of genetically and metabolically engineered strains, optimization of fermentation processes, and extraction and purification processes. The application of multi-omics technologies and computational biology to develop an ectoine producing cell factory was prospected, with the aim to provide a reference for ectoine overproduction.
Amino Acids, Diamino/metabolism* ; Fermentation

Lateral epicondylitis is a common clinical disease with characteristics of lateral elbow pain, insidious onset and easy recurrence, which can cause forearm pain and decreased wrist strength, seriously affecting patients′ daily life and work. Although there are various treatment methods for lateral epicondylitis with different effects, standard treatments are still lacking nowadays. Platelet-rich plasma (PRP) has good effects on bone and tendon repair, and is now widely used in the treatment of lateral epicondylitis. However, there is a lack of a unified understanding of the technology and specifications of PRP in the treatment of lateral epicondylitis. Therefore, the Sports Medicine Branch of the Chinese Medical Association and Physical Medicine and Rehabilitation Branch of the Chinese Medical Association organized experts in the fields of sports medicine and rehabilitation medicine in China to formulate the "clinical expert consensus on platelet-rich plasma treatment for lateral epicondylitis (2022 version)", and proposed suggestions based on evidence-based medicine mainly from the concept, epidemiology and pathophysiology of lateral epicondylitis, symptoms, signs and imaging manifestations of lateral epicondylitis, PRP concept and application component requirements, quality control of PRP preparation technology, indications and contraindications of PRP in the treatment of lateral epicondylitis, PRP injection in the treatment of lateral epicondylitis, application of PRP in the operation of lateral epicondylitis, related problems after PRP treatment of lateral epicondylitis, evaluation of the results after PRP treatment of lateral epicondylitis, and health and economic evaluation of PRP treatment of lateral epicondylitis, so as to provide guidance for clinical diagnosis and treatment.

Objective To establish an animal model of neurogenic pulmonary edema (NPE),and to study the role of catecholamine and beta receptors in the occurrence of NPE. Methods The NPE model was established by injecting fibrinogen and thrombin into the cerebellum medullary pool of the rabbits. Twenty-four rabbits were divided into the control group,the saline group and the experimental group by random num-ber table. In the control group,only cerebellar medullary cistern puncture was carried out,and no drug was in-jected. Cerebrospinal fluid was drew out and the same amount of saline was injected into the cerebellum me-dullary pool in the saline group. Fibrinogen and thrombin were injected into the cerebellum medullary pool in the experimental group. The animals were intubated by tracheotomy,the femoral artery and the internal jugu-lar vein were dissected and connected with the PiCCO instrument to detect the blood pressure,heart rate,and respiratory rate before puncture and at 1 min,10 min,30 min after puncture. Serum samples were collected for the determination of epinephrine,norepinephrine,acetylcholine,endothelin-1,cardiac troponin I,brain natri-uretic peptide and neuropeptide Y levels before puncture and at 1 min,10 min after puncture. The rabbits weresacrificed at 3 hours after successful modeling,the pathological examination of lung was performed. Myocar-dial samples were taken to detect adrenergic beta receptors mRNA. Results (1)The heart rate,respiratory rate and mean arterial pressure at 1 min and 10 min after puncture in experimental group were significantly higher than those in control group and saline group. (2) The pathological examination of the rabbits′ lungs in experimental group found that the lung tissue was swollen and dark red in appearance with large areas of con-gestion. Under the microscope,the lung tissue was edema,bleeding,and inflammatory cells were infiltrated in the alveolar cavity,which was consistent with characteristics of NPE. (3)There was no difference in epineph-rine and norepinephrine concentration in all groups before the cerebellar medullary pool puncture. The concen-tration of epinephrine and norepinephrine at 1 min after puncture time were (200. 0 ± 251. 7)μg/ L,(448. 9 ± 356. 7)μg/ L in the experimental group,whcih were significantly higher than those of control group[(15. 4 ± 3. 4)μg/ L,(15. 9 ± 9. 7)μg/ L] and saline group[(17. 1 ± 3. 8) μg/ L,(29. 6 ± 18. 4) μg/ L] (P < 0. 05). The concentration of epinephrine and norepinephrine at 10 min after puncture were (397. 0 ± 797. 7)μg/ L, (221. 4 ± 173. 7)μg/ L in the experimental group,whcih were significantly higher than those of control group [(23. 3 ± 6. 4) μg/ L,(18. 8 ± 3. 9) μg/ L] and saline group[(16. 7 ± 9. 1) μg/ L,(20. 3 ± 6. 5) μg/ L] (P < 0. 05). (4)There was no significant difference in the levels of serum neuropeptide Y,acetylcholine and endothelin-1 among the three groups. (5)The mRNA of adrenergic beta-1 receptor in the experimental group was 0. 37 ± 0. 12,which was significantly lower than those in the control group (0. 54 ± 0. 13) and saline group (0. 56 ± 0. 14) (P < 0. 05). There was no significant difference in adrenergic beta-3 receptor mRNA among the three groups. Conclusion The NPE animal model was constructed by injecting fibrinogen and thrombin into the cerebellum medullary pool of the rabbits. Catecholamine and beta-1 receptor play a role in the occurrence of NPE rabbit model. There is no significant correlation between neuropeptide Y,acetylcholine,en-dothelin-1 and the occurrence of NPE in rabbits.

Objective: To investigate application and safety of pediatric interfacility-transport with extracorporeal membrane oxygenation (ECMO) in China. Methods: The data of 48 patients transported inter-hospital from February 2016 to May 2018 were collected from the following 4 centers: pediatric intensive care unit (PICU) of Bayi Children′s Hospital Affiliated to the 7th Medical Center of PLA General Hospital, Pediatric Hospital of Fudan University, Henan Provincial People′s Hospital and Children′s Hospital of Zhejiang University School of Medicine. The data of patients′ characteristics, ECMO mode and wean rate, and mortality were reviewed, which was further compared with the data of 57 compatible inner-hospital ECMO cases with t test, Rank sum test or chi-square test. Results: All the 48 interfacility-transports were accomplished by ambulance on land, with an average transfer distance of (435±422) km. The incidence of ECMO complications was 13% (6 case), without death. There were no significant differences in lactic acid, PaO₂ or SaO₂ before and after transport (4.0 (2.0, 7.5) vs. 3.0 (1.5, 6.0) mmol/L, Z=-1.579, P>0.05; 112(47, 405) vs. 166(122, 240) mmHg (1 mmHg=0.133 kPa), Z=-0.104, P>0.05; 0.97±0.02 vs. 0.96±0.03, t=1.570, P>0.05). Instead, PaCO₂ and pH were significantly different ((47±8) vs. (42±5) mmHg, t=2.687, P<0.05; 7.3±0.2 vs. 7.5±0.2, t=3.379, P<0.05). The total ECMO weaned rate was 73% (35/48) and the survival rate was 67% (32/48). No significant differences in demographic characteristics, ECMO mode or duration, transport distance or duration, or complications existed between the survival group and the death group (7/25 vs. 2/14, χ²=0.615, P>0.05; 4/28 vs. 2/14, χ²=0, P>0.05; (405±404) vs. (493±465) km, t=0.525, P>0.05; (5±4) vs. (5±5) h, t=0.388, P>0.05; 166 (128, 239) vs. 187(52, 405) h, Z=-0.104, P>0.05; 3/32 vs. 3/16, χ²=0.734, P>0.05). The lowest lactate value in survival group before ECMO transport was significantly lower than that in the death group ((5±5) vs. (8±6) mmol/L, t=2.151, P<0.05). There were neither significant differences in age, ECMO mode or support pattern (9/39 vs. 15/42, χ²=0.845, P>0.05; 6/42 vs. 7/50, χ²=0.001, P>0.05; 29/19 vs. 38/19, χ²=0.441, P>0.05), nor in ECMO weaned rate, survival rate or complications between interfacility-transport group and inner-hospital group (35/48 vs. 37/57, χ²=0.775, P>0.05; 32/48 vs. 35/57, χ²=0.313, P>0.05; 20/48 vs. 22/57, χ²=0.102, P>0.05). Conclusion With appropriate transport equipment and mature teams who handle problems timely during the transport, critically ill children could be safely transported to the destination with ECMO.

Objective To investigate the changes of cellular immunity and humoral immunity in children with acute or critical illness. Methods Seventy-three critically ill children admitted to PICU of Chil-dren′s Hospital Affiliated to Fudan University during the period from April,2015 to September,2015 were the objects of study. Blood samples were collected within 48 hours after admition. The lymphocyte subpopulation was measured by flow cytometry,and the level of humoral immunity was measured by rate immune scatter turbidimetry,and the neutrophil function was measured by flow cytometry-DHR analysis. Twenty-three cases from 73 cases were detected the second time after admitted to hospital for seven days. Ten health children be-fore elective surgery were selected as control. Results (1) Compared with the control,the percentages of CD3 +T cells and CD8 +T cells were significantly decreased in critically ill children within 48 hours of admis-tion[(57. 43 ± 13. 46)%,(21. 26 ± 7. 87)% vs. (66. 24 ± 5. 27)%,(26. 82 ± 7. 63)%,P<0. 05]; At the same time,CD4 +T cells and NK cells had no significant change[(33. 42 ± 11. 29)%,(8. 83 ± 7. 77)% vs. (34. 89 ± 4. 94)% (11. 34 ± 5. 60)%,both P<0. 05]; While B cells were significantly increased[(31. 69 ±13. 83)% vs. (21. 08 ±7. 24)%,P<0. 05]. Neutrophil activation rate[(14. 32 ±14. 81)%] was signifi-cantly higher than the normal reference value ( 0 -10%) and the activation rate was more than 90% after stimulated by PMA. The plasma level of complement C3[(0. 88 ± 0. 31) g/L] was lower than that of the control group[(1. 19 ± 0. 18)g/L,P<0. 05]. (2) Compared with the first time,the percentages of CD3 +cells and CD4 + cells were increased after treated for one week in 23 patients[(61. 20 ± 13. 56)%,(36. 79 ± 9. 95)% vs. (56. 80 ± 13. 99)%,(32. 86 ± 10. 87)%,both P<0. 05]. No significant difference in neutrophil activation and activation rate after PMA stimulation was found compared with admition. IgA,IgM and comple-ment C3 were significantly increased[(0. 98 ± 0. 75) g/L,(1. 00 ± 0. 39) g/L,(1. 15 ± 0. 34) g/L vs. (0. 80 ± 0. 69) g/L,(0. 86 ± 0. 48) g/L,(0. 93 ± 0. 23) g/L,all P<0. 05]. Conclusion Immune disorders occur in critically ill children in the early stage of illness,the most obvious change is cellular immune response,and im-mune function starts to recover after one week.

Objective To study the Hainan Utstein templates used for cardiac arrest and resuscitation registries to evaluate the epidemiological characteristics and outcomes of the patients with CPR by multi-center study. Methodsccording to the Utstein templates for cardiac arrest and CPR set by International Liaison Committee on resuscitation in 2004, a Hainan Utstein CPR registry chart was designed and a prospective descriptive study was carried out to evaluate the epidemiological characteristics, impact factors and outcomes of the patients with resuscitation attempt in emergency departments of thirteen hospitals in Hainan Island between January 2007 and December 2010.Results Of 1125 patients with cardiac arrest, male accounted for 73. 8％ and female was 26. 2％. The mean ( ± S. D) age of the cardiac arrest patients was 53.9 ± 13. 1 years old.Coronary heart diseases and hypertension were the most common preexisting chronic diseases in the studied patients. The ROSC rate and discharge rates after survival in 1125 patients with CPR were 23. 8％ and 7.4％ respectively. The ROSC rate and discharge rates after survival were 36. 3％ and 11.6％ in the in-hospital cardiac arrest (IHCA) group, respectively whereas 11.5％ and 3. 3％ in out-hospital cardiac arrest (OHCA) group. Of 188 patients with ventricular fibrillation/Pulseless ventricular tachycardia, the ROSC rate and discharge rate after survival were 58.0％and 21.8％,respectively. Of them, 448 (39. 8％ ) of the cardiac arrest patients had underlying cardiac causes, and the ROSC rate and discharge rate after survival were 36. 3％ and 11.5％ respectively in IHCA group whereas 11.6％ and 3. 3％ in OHCA group. The ROSC rate and discharge rate after survival were 69. 8％ and 7. 4％respectively in the tertiary hospitals whereas 30. 2％ and 7. 3％ in the secondary hospitals. Conclusions Patients experienced cardiac arrest were predominantly male. Coronary heart disease and hypertension were the two most common preexisting chronic diseases. The ROSC rate and discharge rate of patients with IHCA were higher than those with OHCA. ROSC rate and discharge rate after survival were higher in the ventriculat fibrillation/Pulseless ventricular tachycardia group than the other cardiac rhythms first witnessed groups. The time delayed of starting CPR after onset of cardiac arrest had a critical impact on survival and discharge rate in both IHCA and OHCA groups.

Objective To analyze the electrophysiologie characteristics of critical illness of polyneuromyopathy ( CIPM ) and its effect on the duration of usage of mechanical ventilation. Methods Denmark Keypoint 4-electromyogram was utilized to record electraneurophysiology on 17 subjects with lung infection and respiratory failure who were treated with mechanical ventilation over 1 week and had difficulty in weaning from the ventilator in intensive care unit. Electrophysiology character of CIPM was described and the duration of mechanical ventilation of CIPM patients was compared with non-CIPM subjects. ResultsOf the 17 cases, 9 subjects were diagnosed with CIPM including 6 patients with critical illness polyneuropathy (CIP), and 3 patients with critical illness myopathy (CIM), the 2 subtype of CIPM. Compound muscle action potential (CMAP) and sympathetic skin response (SSR) in all of CIPM patients were abnormaL Though the amplitude of sensory nerve action potential (SNAP) in patients with CIP was reduced, nerve conduction velocity (NCV) was normal in all of them. Repetitive nerve stimulation, somatosensory evoked potential and blink reflex were normal in all of the patients with CIPM, however, the rate of occurrence of F wave was reduced in 2 patients with CIP and in 2 patients with CIM. The median and quartile range of duration of mechanical ventilation in CIPM patients was 30. 0 days (20. 0--45.0 days), significantly longer than13.5 days (9.5--17.5 days) in non-CIPM patients (U=7.500,P=0.006). ConclusionsThe ehctrophysiology change in patients with CIPM is characterized by reduced amplitude of CMAP and (or) SNAP, normal NCV and abnormal SSR. The duration of mechanical ventilation is significantly prolonged in CIPM patients.