Objective:To establish a rapid quantitative PCR (qPCR) technique for Mycobacterium marinum skin infections, and to analyze its clinical diagnostic efficiency. Methods:DNA was extracted from Mycobacterium marinum colonies and serially diluted (10 -1 to 10 -8). Twelve pairs of previously reported primers and probes, as well as 6 pairs of newly designed primers and probes in this study, were used for qPCR amplification to identify the most sensitive primers and probes for the detection of Mycobacterium marinum. Skin lesion tissues were collected from 72 patients with confirmed Mycobacterium marinum infections (experimental group) and 68 with other mycobacterial infections (control group) at Shandong Provincial Hospital for Skin Diseases & Shandong Provincial Institute of Dermatology and Venereology, Shandong First Medical University & Shandong Academy of Medical Sciences in 2021. These skin tissues were subjected to qPCR amplification, interferon-gamma release assay (IGRA), acid-fast staining, and tissue culture to evaluate the diagnostic efficacy. Results:The newly designed primers and probes targeting the mycobacterial enhanced infection locus 2 (Mel2) demonstrated the highest sensitivity, with a detection limit of 0.86 copies/μl (cycle threshold value = 37) ; the qPCR amplification with the Mel2 primers/probes did not yield positive results when used for the detection of other mycobacteria (including Mycobacterium leprae and Staphylococcus spp) . Among the 72 patients in the experimental group, 44 were positive for qPCR with a sensitivity of 61.1% (95% CI: 49.6% - 71.5%), and 47 were positive for culture with a sensitivity of 65.2% (95% CI: 53.8% - 75.3%) ; all the 68 controls were negative for both qPCR and culture, with their specificities both being 100%. Among 65 patients subjected to IGRA, 31 were positive with a sensitivity of 47.7% (95% CI: 36.0% - 59.6%), while 16 out of 25 controls were negative for IGRA with a specificity of 64.0% (95% CI: 44.5% - 79.8%). Among 58 patients subjected to acid-fast staining, 37 were positive with a sensitivity of 63.8% (95% CI: 50.9% - 74.9%), and 52 out of 66 controls were negative for acid-fast staining with a specificity of 78.8% (95% CI: 67.5% - 86.9%). The combination of qPCR and culture resulted in a sensitivity of 93% and a specificity of 100% for the detection of Mycobacterium marinum. Conclusion:In this study, a highly sensitive qPCR assay was developed for the detection of Mycobacterium marinum, and its combination with culture could further improve the detection sensitivity.

Itaconate is a pivotal intermediate metabolite in the tricarboxylic acid (TCA) cycle of immune cells. It is produced by decarboxylation of cis-aconitic acid under the catalysis of aconitate decarboxylase 1 (ACOD1), which is encoded by the immune response gene 1 (IRG1). Itaconate has become a focal point of research on immunometabolism. Studies have demonstrated that itaconate plays a crucial role in diseases by regulating inflammation, remodeling cell metabolism, and participating in epigenetic regulation. This paper reviewed the research progress in itaconnate from its chemical structure, regulatory effects on different diseases, and mechanisms, proposes the future research directions, aiming to provide a theoretical basis for the development of itaconate-related drugs.
Humans ; Succinates/metabolism* ; Carboxy-Lyases/genetics* ; Inflammation/metabolism* ; Citric Acid Cycle ; Animals ; Epigenesis, Genetic ; Neoplasms/immunology*

Ten-eleven translocation 1 (TET1) protein is an alpha-ketoglutaric acid (α-KG) and Fe²⁺-dependent dioxygenase. It plays a role in the active demethylation of DNA by hydroxylation of 5-methyl-cytosine (5-mC) to 5-hydroxymethyl-cytosine (5-hmC). Ten-eleven translocation 1 (TET1) protein is involved in maintaining genome methylation homeostasis and epigenetic regulation. Abnormally expressed TET1 and 5-mC oxidative derivatives have become potential markers in various biological and pathological processes and a research focus in the fields of embryonic development and malignant tumors. This paper introduces the structure and demethylation mechanism of TET1, reviews the research status of epigenetic regulation by TET1 in embryonic development, immune responses, stem cell regulation, cancer progression, and nervous system development, and briefs the upstream regulatory mechanism of TET1, hoping to provide new inspirations for further research in related fields.
Proto-Oncogene Proteins/genetics* ; Epigenesis, Genetic ; Humans ; DNA-Binding Proteins/metabolism* ; DNA Methylation ; Mixed Function Oxygenases/metabolism* ; 5-Methylcytosine/analogs & derivatives* ; Animals ; Embryonic Development/genetics* ; Neoplasms/genetics* ; Dioxygenases/metabolism*

The chronic use of morphine and other opioids is associated with opioid-induced hypersensitivity (OIH) and analgesic tolerance. Among the different forms of OIH and tolerance, the opioid receptors and cell types mediating opioid-induced mechanical allodynia and anti-allodynic tolerance remain unresolved. Here we demonstrated that the loss of peripheral μ-opioid receptors (MORs) or MOR-expressing neurons attenuated thermal tolerance, but did not affect the expression and maintenance of morphine-induced mechanical allodynia and anti-allodynic tolerance. To confirm this result, we made dorsal root ganglia-dorsal roots-sagittal spinal cord slice preparations and recorded low-threshold Aβ-fiber stimulation-evoked inputs and outputs in superficial dorsal horn neurons. Consistent with the behavioral results, peripheral MOR loss did not prevent the opening of Aβ mechanical allodynia pathways in the spinal dorsal horn. Therefore, the peripheral MOR signaling pathway may not be an optimal target for preventing mechanical OIH and analgesic tolerance. Future studies should focus more on central mechanisms.
Humans ; Morphine/pharmacology* ; Hyperalgesia/metabolism* ; Analgesics, Opioid/pharmacology* ; Neurons/metabolism* ; Signal Transduction

ObjectiveTo establish the specific chromatogram and thin layer chromatography（TLC） identification method of Kaixinsan（KXS） samples， in order to clarify the key quality attributes and provide reference for the quality evaluation of KXS. MethodHigh performance liquid chromatography（HPLC） specific chromatogram of KXS was developed with YMC Hydrosphere C₁₈ column（4.6 mm×250 mm， 5 μm）， the mobile phase was acetonitrile（A）-0.2% formic acid aqueous solution（B） for gradient elution（0-15 min， 2%-20%A； 15-25 min， 20%-25%A； 25-30 min， 25%-30%A； 30-45 min， 30%-31%A； 45-50 min， 31%-44%A； 50-65 min， 44%-45%A； 65-73 min， 45%-75%A； 73-95 min， 75%-100%A； 95-105 min， 100%A； 105-105.1 min， 100%-2%A； 105.1-120 min， 2%A）， the detection wavelength was 320 nm. Ultra high performance liquid chromatography-linear ion trap-electrostatic field orbitrap mass spectrometry（UHPLC-LTQ-Orbitrap MS） was used to identify the chemical components of KXS with electrospray ionization（ESI）， negative ion mode and scanning range of m/z 50-2 000. TLC identification methods for Poria and Ginseng Radix et Rhizoma in KXS were established. ResultThere were 11 common peaks in the specific chromatogram of KXS， attributed to Polygalae Radix， Poria and Acori Tatarinowii Rhizoma. Taking peak 9（α-asarone） as the reference peak， the relative standard deviations of the retention times of 15 batches of KXS samples were<0.2%. A total of 34 compounds were identified by UHPLC-LTQ-Orbitrap MS， including terpenoids， phenylpropanoids， oligosaccharides and ketones. The established TLC had good separation and was rapid， reliable， simple， feasible， suitable for the identification of Poria and Ginseng Radix et Rhizoma in KXS. ConclusionThe specific chromatogram and TLC of KXS are stable and reproducible. The material basis of KXS is basically clarified by MS， which can provide a reference for the development and quality control of KXS.

Objective:To evaluate the effect of dexmedetomidine on perioperative neurocognitive disorders in elderly frail patients undergoing hip joint surgery.Methods:Sixty elderly patients of either sex, aged≥ 60 yr, weighing 40-100 kg, of American Society of Anesthesiologists Physical Status classification Ⅰ-Ⅲ, with the Frail Scale score 3-5 points, scheduled for elective hip surgery under spinal-epidural anesthesia, were divided into 2 groups ( n=30 each) using the random number table method: control group (group C) and dexmedetomidine group (group D). Dexmedetomidine was intravenously infused at a dose of 0.5 μg/kg at 10 min before anesthesia followed by a continuous infusion of 0.2 μg·kg -1·h -1 until 10 min before the end of surgery in group D. The equal volume of 0.9% sodium chloride solution was given at the corresponding time point in group C. Blood samples from the median cubital vein were collected before surgery (T 1) and at 1 and 3 days after surgery (T 2, 3) for determination of concentrations of serum S100β and neuron-specific enolase by enzyme-related immunosorbent assay. Postoperative delirium was assessed within 3 days after surgery using the Confusion Assessment Method. Cognitive function was evaluated by Mini-Mental State Examination at T 1 and 30 days after surgery. Results:Compared with the baseline at T 1, the concentrations of serum S100β and NSE were significantly increased at T 2 and T 3 in two groups ( P<0.05). Compared with group C, the concentrations of serum S100β and neuron-specific enolase and incidence of postoperative delirium and postoperative cognitive dysfunction were significantly decreased at T 2 and T 3 in group D( P<0.05). Conclusions:Dexmedetomidine can effectively decrease the occurrence of perioperative neurocognitive disorders in elderly frail patients undergoing hip joint surgery.

Objective:To investigate the relationship between carotid atherosclerosis(CAS)and subclinical left ventricular(LV)dysfunction in type 2 diabetes mellitus patients with preserved LV ejection fraction(LVEF).Methods:A total of 120 patients with type 2 diabetes mellitus who had LVEF≥50% were selected in the Department of Endocrinology, the First Affiliated Hospital of Air Force Medical University from June 2021 to October 2021. The global longitudinal strain(GLS)was obtained by two-dimensional speckle tracking echocardiography(STE)to assess subclinical LV systolic function. The mitral ratio of peak early to late diastolic filling velocity(E/A), and mitral velocity to early diastolic velocity of the mitral annulus(E/E′)ratio were obtained by pulsed tissue Doppler echocardiography to assess LV diastolic function. Acrroding to bilateral carotid ultrasound examination, the subjects were divided into normal carotid arteries group( n=46) and CAS group( n=74). Demographics and biochemical parameters were compared between two groups. Binary logistic regression and Pearson correlation analysis were used to evaluate the relationship between CAS and subclinical LV dysfunction. Results:The CAS group had a higher proportion of men, older age, and a longer duration of diabetes than the normal carotid arteries group(all P<0.05). There was no difference in LVEF and GLS between the two groups [normal carotid arteries group vs CAS group, LVEF: (60.72±4.73)% vs(60.07±4.28)%; GLS: (18.24±3.72)% vs(17.81±3.47)%, respectively; both P>0.05]. However, compared with normal carotid arteries group, E/A ratio was decreased and E/E′ ratio was significantly increased in CAS group(both P<0.01). Pearson correlation analysis showed that GLS was not correlated with carotid plaque thickness and carotid intima-media thickness(CIMT; both P>0.05). By contrast, E/E′ ratio was positively correlated with carotid plaque thickness and CIMT(both P<0.05). Binary logistic regression analysis showed that GLS and E/E′ ratio were not associated with CAS( both P>0.05). However, decreased E/A ratio was significantly associated with the existence of CAS( OR=0.09, 95% CI 0.01-0.67, P=0.018). Conclusions:In type 2 diabetes mellitus patients without overt heart failure and with preserved LVEF, the occurrence of CAS is not associated with subclinical LV systolic impairment assessed by GLS, but is significantly associated with LV diastolic dysfunction, and is independent of traditional cardiovascular risk factors.

Objective:To investigate the influencing factors of non-remission of proteinuria in patients with nephrotic syndrome (NS) and idiopathic membranous nephropathy (IMN).Methods:The study was a retrospective observational study. The clinical data of patients with NS who were diagnosed as IMN by renal biopsy and serum albumin recovered normal after six months of treatment were collected from Beijing Anzhen Hospital, Capital Medical University from June 1, 2010 to January 31, 2022. Patients were divided into proteinuria remission group and non-proteinuria remission group according to whether urinary protein < 3.5 g/24 h and decreased 50% from the onset. The differences of clinical and pathological characteristics between the two groups at baseline were compared. The logistic regression model was used to analyze the influencing factors of non-remission of proteinuria.Results:Ninety-five NS patients with renal pathology of IMN were included in this study, with age of 57(43, 65) years old and 50 males (52.6%). There were 75 patients in the proteinuria remission group and 20 patients in the non-proteinuria remission group. Compared with the proteinuria remission group, the non-proteinuria remission group had higher baseline body mass index [(26.83±4.03) kg/m 2vs. (24.68±3.97) m 2, t=-2.149, P=0.034] and proportion of overweight (85.0% vs. 58.7%, χ2=4.765, P=0.029), and larger waist circumference [88.5(85.3, 101.5) cm vs. 87.0(77.5, 92.0) cm, Z=2.362, P=0.018]. Renal pathological results showed that the proportions of diabetes nephropathy (10.0% vs. 0, P=0.043) and glomerular hypertrophy (45.0% vs. 20.0%, χ2=5.227, P=0.022) were higher, and the average diameter of hypertrophic glomeruli was longer [(197.96±6.37) μm vs. (193.51±8.50) μm, t=2.029, P=0.041] in the proteinuria remission group than those in the non-proteinuria remission group. Multivariate logistic regression analysis results showed that waist circumference was an independent influencing factor of non-proteinuria remission in patients with IMN under waist circumference > 90 cm in men and >85 cm in women ( OR=1.083, 95% CI 1.005-1.168, P=0.037). Conclusion:Abdominal obesity is an independent risk factor of non-remission of proteinuria in NS patients with IMN after early treatment.

Objective:To investigate the relationship between spinal Mas-related gene receptor C (MrgC) and pathophysiological mechanism of bone cancer pain in mice.Methods:Forty male C3H/HeNCrlVr mice, aged 5-7 weeks, weighing 20-25 g, were selected and divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group S), bone cancer pain group (group P), bone cancer pain + MrgC agonist group (group P-agonist) and bone cancer pain + Mrg C antagonist group (group P-Ab). Preparation of the bone cancer pain model: mouse fibrosarcoma cells (NCTC2472) were injected into the upper tibia of mice in P, P-agonist and P-Ab groups, and the equal volume of D-Hanks balanced salt solution was given instead in S group. Fourteen days later cerebrospinal fluid was intrathecally injected in S and P groups, and MrgC agonist and MrgC antibody were intrathecally injected in P-agonist and P-Ab groups. The mechanical paw withdrawal threshold (MWT) to von Frey stimuli was measured before developing the model (T 0), at 7 days after developing the model (T 1), at 14 days after developing the model (before intrathecal injection, T 2), and at 4, 8 and 12 h after intrathecal injection (T 3-5). Results:Compared with group S, no significant change was found in the MWT at T 0 ( P>0.05), and the MWT was significantly decreased at T 1-T 5 in the other groups ( P<0.05). Compared with group P, the MWT was significantly increased at T 3-T 5 in group P-agonist, and the MWT was significantly decreased at T 3-T 5 in group P-Ab ( P<0.05). Conclusions:Spinal MrgC plays an endogenous protective role in the pathophysiological mechanism of bone cancer pain to a certain extent in mice.

Objective:To analyze the diagnostic and prognostic value of routine bone marrow examination in patients with extranodal NK/T-cell lymphoma (ENKTCL) based on PET-CT staging.Methods:Clinical data of 186 patients who received bone marrow biopsy and bone marrow aspiration in Fujian Medical University Union Hospital from 2013 to 2021 were retrospectively analyzed. All patients were divided into bone marrow biopsy + bone marrow aspiration group ( n=186) and PET-CT + bone marrow biopsy group ( n=139). The sensitivity, specificity, positive and negative predictive values were compared between two groups. The data were analyzed and plotted. Survival analysis was performed using Kaplan-Meier method and log-rank test. Results:In the whole cohort, 45 patients were positive for bone marrow biopsy, and 30 of them were positive for bone marrow aspiration. A total of 141 patients who were negative for bone marrow biopsy also achieved negative results for bone marrow aspiration. A total of 139 patients completed PET-CT staging and bone marrow biopsy. And 30 patients were diagnosed with positive bone marrow by PET-CT, in which 22 of them were confirmed positive by bone marrow biopsy. Among 109 patients diagnosed with negative bone marrow by PET-CT, 5 of them were confirmed positive by bone marrow biopsy. All these cases were classified as stage Ⅳ due to distant metastases. PET-CT had a diagnostic sensitivity of 81.5%, a specificity of 92.9%, a positive predictive value of 73.3%, and a negative predictive value of 95.4%. Among early stage (Ⅰ-Ⅱ stage) patients diagnosed with PET-CT, all of them were negative for bone marrow biopsy (the negative predictive value was 100%). In stage Ⅳ patients ( n=55), the 1-year overall survival of patients with bone marrow involvement by bone marrow biopsy or PET-CT ( n=35) compared with their counterparts with the involvement of other organs ( n=20) was 28.7% vs.42.0% ( P=0.13), and 1-year progression free survival rates was 23.2% vs. 23.3% in ( P=0.94). Conclusions:Routine bone marrow biopsy does not change the original staging of patients with early stage ENKTCL based on PET-CT staging. Advanced stage patients with positive bone marrow biopsy tend to obtain worse prognosis, indicating that bone marrow biopsy still has certain value.