Objective:To explore the effect of exosomes released by Mycobacterium tuberculosis ( Mtb) Rv1983-stimulated macrophages on tuberculosis infection and its potential mechanism. Methods:Exosomes (Rv1983-M-Exo) released by macrophages following Mtb Rv1983 stimulation were extracted by hypervelocity centrifugation and then co-cultured with uninfected macrophages. Macrophage activity was detected by prodium iodide (PI) staining. The level of lipid reactive oxygen species (ROS) was detected by lipid peroxidation fluorescence probe. The expression of ferrous ions (Fe 2+ ) and malondialdehyde (MDA) were detected by colorimetry assay. The protein expression of acyl-CoA synthetase long chain family member 4 (ACSL4) in Rv1983-M-Exo was detected by Western blot. Exosomes (Rv1983-M+ siACSL4-Exo) were isolated from Rv1983-stimulated macrophages with interfered ACSL4 expression Rv1983 and co-cultured with uninfected macrophages. The effect of exosomes on the polarization of macrophages was detected by flow cytometry and Western blot. The effects of exosomes on phagocytosis and killing ability of macrophages were analyzed by plate colony assay and BCG-phagocytosis lysosome co-localization assay. Results:Compared with macrophage-derived exosomes (M-Exo), Rv1983-M-Exo promoted ferroptosis in uninfected macrophages, manifested by increased levels of intracellular lipid ROS, MDA and Fe 2+, which were significantly inhibited by ferroptosis inhibitor Fer-1. Rv1983 induced macrophages to release exosomes with high expression of ACSL4. Interfering the expression of ACSL4 in macrophages, the concentration of ACSL4 in the Rv1983-M+ siACSL4-Exo was significantly reduced. When the Rv1983-M+ siACSL4-Exo was co-cultured with uninfected macrophages, the ferroptosis of macrophages was significantly reduced. Rv1983-M-Exo promoted M2 macrophage polarization which showed up-regulation of CD206 and Arg1 expression, and decreased the phagocytosis and killing ability of macrophages, while Rv1983-M+ siACSL4-Exo or Fer-1 in combination with Rv1983-M-Exo reduced the expression of CD206 and Arg1 in macrophages, and promoted the phagocytosis and killing ability. Conclusions:Mtb Rv1983 protein up-regulates the level of ACSL4 in the exosomes secreted by macrophages, thereby inducing ferroptosis in uninfected macrophages, causing M2 polarization of macrophages, weakening phagocytosis and killing ability, and promoting Mtb infection.

ObjectiveTo investigate the effects of thyme herbal tea （BLX） on the proliferation of human coronavirus OC43 （HCoV-OC43） in vitro and in vivo. MethodThe chemical composition of BLX was analyzed by UPLC-MS. The cytotoxicity of BLX in HRT-18 cells and the effect of BLX treatment on the proliferation of HCoV-OC43 in cells were analyzed. Copies of viral gene were detected by real-time PCR. The effect of BLX treatment on the life cycle of HCoV-OC43 was detected by time-of-addition assay. The maximum tolerated dose of BLX and the influences of BLX on the body weight and survival time of suckling mice infected with HCoV-OC43 were determined. The expression of viral protein in the brain and lung tissue was analyzed by immunohistochemistry. ResultThere were 11 chemical components identified in BLX by UPLC-MS. BLX showed the 50% cytotoxic concentration （CC₅₀） of （13 859.56±319） mg·L^-1， the median inhibitory concentration （IC₅₀） of （1 439.09±200） mg·L^-1， and the selection index of 8.26-11.44 for HCoV-OC43 in HRT-18 cells. Compared with the cells infected with HCoV-OC43， BLX at the concentrations of 1 500， 1 000， 500 mg·L^-1 inhibited the proliferation of this virus （P<0.05， P<0.01）. BLX exhibited antiviral effect in the early stage of virus infection， and the inhibition role in the attachment stage was more significant than that in the entry stage （P<0.05）. In the suckling mice infected with HCoV-OC43， BLX at 1200 and 600 mg·kg^-1·d^-1 alleviated the symptoms， prolonged the survival period， reduced the death rate， and down-regulated the mRNA level of nucleocapsid protein in the mice. Moreover， BLX at 1 200 mg·kg^-1·d^-1 down-regulated the expression of nucleocapsid protein in the brain （P<0.01） and the lung （P<0.01）. ConclusionBLX contained multiple antiviral ingredients. It inhibited the proliferation of HCoV-OC43 both in vitro and in vivo by interference with viral attachment. This study provides theoretical reference for the treatment of acute respiratory tract infection with HCoV-OC43 and for further development and application of BLX.

Objective:To investigate the Helicobacter pylori ( H. pylori) infection of Tibetan families and individuals in the Western Sichuan Plateau region and explore the related factors which affected H. pylori infection. Methods:This was a single-center cross-sectional study. Questionnaires were collected from 50 Tibetan families including 155 individuals in Western Sichuan Plateau region during March to May 2023. The 13C-urea breath test was performed to confirm the current infection status of participants. Binary logistic regression were used to analyze the related factors associated with H. pylori infection. Results:Among the 50 Tibetan households, the individual-based H. pylori infection rate was 47.10%(73/155), with two out of nine children and 48.63%(71/146) adults infected. The age group of 18 to 40 years had the highest infection rate (55.00%, 11/20). The prevalence of infection based on family was 80.00%(40/50), of which 16.00%(8/50) had all family members infected. Of the 59 couples surveyed, 23.73%(14/59) were both infected, and 45.76%(27/59) had one person infected. Of the six families which had children and adolescents, two households had their children infected. Logistic regression analysis showed that size of the family was a factor related to H. pylori infection (odds ratio=3.038, 95% confidence interval 1.043 to 10.491, P=0.042). Conclusions:The family-based H. pylori infection rate is relatively high in Tibetan residents in the Western Sichuan Plateau, and larger family size is related with higher risk of H. pylori infection within the family.

Objective:To explore the effect of exosomes released by Mycobacterium tuberculosis ( Mtb) Rv1983-stimulated macrophages on tuberculosis infection and its potential mechanism. Methods:Exosomes (Rv1983-M-Exo) released by macrophages following Mtb Rv1983 stimulation were extracted by hypervelocity centrifugation and then co-cultured with uninfected macrophages. Macrophage activity was detected by prodium iodide (PI) staining. The level of lipid reactive oxygen species (ROS) was detected by lipid peroxidation fluorescence probe. The expression of ferrous ions (Fe 2+ ) and malondialdehyde (MDA) were detected by colorimetry assay. The protein expression of acyl-CoA synthetase long chain family member 4 (ACSL4) in Rv1983-M-Exo was detected by Western blot. Exosomes (Rv1983-M+ siACSL4-Exo) were isolated from Rv1983-stimulated macrophages with interfered ACSL4 expression Rv1983 and co-cultured with uninfected macrophages. The effect of exosomes on the polarization of macrophages was detected by flow cytometry and Western blot. The effects of exosomes on phagocytosis and killing ability of macrophages were analyzed by plate colony assay and BCG-phagocytosis lysosome co-localization assay. Results:Compared with macrophage-derived exosomes (M-Exo), Rv1983-M-Exo promoted ferroptosis in uninfected macrophages, manifested by increased levels of intracellular lipid ROS, MDA and Fe 2+, which were significantly inhibited by ferroptosis inhibitor Fer-1. Rv1983 induced macrophages to release exosomes with high expression of ACSL4. Interfering the expression of ACSL4 in macrophages, the concentration of ACSL4 in the Rv1983-M+ siACSL4-Exo was significantly reduced. When the Rv1983-M+ siACSL4-Exo was co-cultured with uninfected macrophages, the ferroptosis of macrophages was significantly reduced. Rv1983-M-Exo promoted M2 macrophage polarization which showed up-regulation of CD206 and Arg1 expression, and decreased the phagocytosis and killing ability of macrophages, while Rv1983-M+ siACSL4-Exo or Fer-1 in combination with Rv1983-M-Exo reduced the expression of CD206 and Arg1 in macrophages, and promoted the phagocytosis and killing ability. Conclusions:Mtb Rv1983 protein up-regulates the level of ACSL4 in the exosomes secreted by macrophages, thereby inducing ferroptosis in uninfected macrophages, causing M2 polarization of macrophages, weakening phagocytosis and killing ability, and promoting Mtb infection.

The Ly-6 and uPAR (LU) domain-containing proteins represent a large family of cell-surface markers. In particular, mouse Ly-6A/Sca-1 is a widely used marker for various stem cells; however, its human ortholog is missing. In this study, based on a systematic survey and comparative genomic study of mouse and human LU domain-containing proteins, we identified a previously unannotated human gene encoding the candidate ortholog of mouse Ly-6A/Sca-1. This gene, hereby named LY6A, reversely overlaps with a lncRNA gene in the majority of exonic sequences. We found that LY6A is aberrantly expressed in pituitary tumors, but not in normal pituitary tissues, and may contribute to tumorigenesis. Similar to mouse Ly-6A/Sca-1, human LY6A is also upregulated by interferon, suggesting a conserved transcriptional regulatory mechanism between humans and mice. We cloned the full-length LY6A cDNA, whose encoded protein sequence, domain architecture, and exon-intron structures are all well conserved with mouse Ly-6A/Sca-1. Ectopic expression of the LY6A protein in cells demonstrates that it acts the same as mouse Ly-6A/Sca-1 in their processing and glycosylphosphatidylinositol anchoring to the cell membrane. Collectively, these studies unveil a novel human gene encoding a candidate biomarker and provide an interesting model gene for studying gene regulatory and evolutionary mechanisms.
Humans ; Membrane Proteins/genetics* ; Pituitary Neoplasms/genetics* ; Biomarkers

Objective:To systematically evaluate the effects of a low-microbiological diet on the incidence of infection in cancer patients with agranulocytosis.Methods:The Cochrane Library, PubMed, Embase, Web of Science, CNKI, China Biology Medicine disc (CBM) , Wanfang Database and VIP database were searched by computer. After two researchers independently carried out literature screening, data extraction and evidence quality evaluation, RevMan 5.3 software was used for Meta-analysis.Results:A total of 7 studies were included, including 5 RCT studies and 2 retrospective case-control trials. Meta-analysis results showed that in retrospective controlled studies [ RR=1.29, 95% CI (1.10, 1.50) , P=0.002], and between different subjects (transplanted and non-transplanted patients) [ RR=1.21, 95% CI (1.08, 1.36) , P=0.001], the incidence of infection in the routine diet group was lower than that in the low-microbial diet group. In the prospective randomized controlled study, there was no statistically significant difference in the infection rate between the two groups [ RR=1.08, 95% CI (0.92, 1.28) , P=0.340]. In terms of infection-related mortality, there was no statistically significant difference between the two groups [ RR=1.06, 95% CI (0.76, 1.47) , P=0.750]. Conclusions:Compared with patients with unrestricted diet, the low-microbial diet does reduce the infection rate and infection-related mortality of patients. Dietary restrictions for patients with granulocytic cancer may be appropriately relaxed, and a focus on safe food purchase and handling and good hand hygiene may provide better infection prevention for patients with agranulocytosis.

Objective:To explore the effects of sequential application of intensive pulsed light and carbon dioxide laser in treating the hypertrophic scars of burn children at early stage.Methods:A retrospective cohort before-after control study in the same patients was conducted. From January 2016 to December 2018, 145 burn children with hypertrophic scar at the early stage who met the inclusion criteria were admitted to the First Hospital of Jilin University, including 82 males and 63 females, aged 1 to 12 (3 (2, 6)) years. All the children were firstly treated with intense pulsed light therapy (no anesthesia or intravenous-inhalation combined anesthesia) at an interval of once per month, and then changed to carbon dioxide laser therapy (topical anesthesia or intravenous-inhalation combined anesthesia) when the degree of scar hyperemia was reduced, at an interval of once every 3 months, for a total of 3 times. Before the first intense pulsed light treatment (hereinafter referred to as before the first treatment) and 3 months after the last carbon dioxide laser treatment (hereinafter referred to as after the last treatment), scar scoring was evaluated by Vancouver Scar Scale (VSS), and scar hyperemia (denoted as hemoglobin level) was measured with Antera 3D ? camera. The times of intense pulsed light, the time of single treatment, the anesthesia method, and the time of intravenous-inhalation combined anesthesia of intense pulsed light and carbon dioxide laser treatment were analyzed. After the last treatment, Likert Scale was used to evaluate the efficacy satisfaction of both doctors and patients. Adverse reactions were recorded during the treatment. Data were statistically analyzed with Wilcoxon signed rank sum test, and paired sample t test. Results:The color, vascular distribution, thickness, and softness scores, and total score in VSS scoring of scars of children after the last treatment were significantly lower than those before the first treatment ( Z=-6.05, -10.34, -9.84, -9.28, -10.43, P<0.01). The hemoglobin level of scar of children after the last treatment was 1.86±0.24, significantly lower than 2.27±0.32 before the first treatment ( t=17.65, P<0.01). A total of 411 times of intense pulsed light therapy were performed, (2.8±0.6) times per person, and the single treatment time was 35 (20, 45) s. There were 392 times (95.38%) without anesthesia, and 19 times (4.62%) with intravenous-inhalation combined anesthesia with time of 6 (5, 8) min. The single treatment time of carbon dioxide laser therapy was 5 (3, 10) min. There were 364 times (83.68%) of topical anesthesia and 71 times (16.32%) of intravenous-inhalation combined anesthesia with time of 10 (8, 15) min. After the last treatment, the efficacy satisfaction scores of doctors and patients were (4.3±0.7) and (3.8±1.0) points, respectively. Blisters occurred in 5 cases after intense pulsed light treatment, which were healed naturally after drainage. One child developed local skin infection, skin redness and swelling accompanied by purulent exudate after carbon dioxide laser treatment, which was improved after skin disinfection and external use of mupirocin ointment. No inflammatory pigmentation, worsening of hyperplasia of scar, erythema, or other skin adverse reactions or anesthetics-related adverse reactions occurred in any child. Conclusions:Sequential application of intense pulsed light and carbon dioxide laser to treat the hypertrophic scars of burn children at early stage can obviously improve the appearance and texture of scar, with higher satisfaction of doctors and patients and fewer adverse reactions.

Objective:To investigate the value of α-synuclein (α-syn) concentration in cerebrospinal fluid (CSF) in predicting postoperative delirium (POD).Methods:One thousand patients underwent elective surgery with combined epidural-spinal anesthesia in our hospital from January 2018 to September 2020 were selected.The epidural puncture was performed at L 3, 4 interspace, and 2 ml of CSF was collected after the needle reaching the subarachnoid space.The concentrations of α-syn, β-amyloid (Aβ)40, Aβ42, total tau protein (T-tau), and phosphorylated tau protein (P-tau) in CSF were determined by enzyme-linked immunosorbent assay.The concentrations of α-syn in CSF and occurrence of POD in patients of different ages were recorded.Patients were divided into POD group and non-POD group according to whether POD occurred, and frequency matching (1∶1) was performed based on five matching variables of age, ASA physical status, education level, duration of operation, and intraoperative blood loss. Results:Eight hundred and forty-one patients were finally included in the study, and the incidence of POD was 15.0%. There were 126 cases in POD group and 126 cases in non-POD group after matching. The concentrations of α-syn in CSF and incidence of POD were gradually increased with age ( P<0.05). Compared with non-POD group, the concentrations of α-syn, T-tau and P-tau in CSF were significantly increased, the concentrations of Aβ40 and Aβ42 were decreased, Aβ40/P-tau, Aβ42/P-tau, Aβ42/Aβ40 and P-tau/T-tau were decreased in POD group ( P<0.05). After confounding factors were corrected by logistic regression analysis, increased concentrations of α-syn, p-tau, and T-tau in CSF were risk factors for POD ( P<0.05). Increased concentrations of Aβ40 and Aβ42 in CSF and increased Aβ40/P-tau and Aβ42/P-tau were protective factors for POD ( P<0.05). Multiple linear regression analysis showed that the concentration of α-syn in CSF was negatively correlated with Aβ40 and Aβ42 concentrations and positively correlated with P-tau and T-tau concentrations ( P<0.05). The area under the receiver operating characteristic curve of concentrations of α-syn in CSF predicting POD was 0.895, Youden index was 0.664, sensitivity was 80.00%, and specificity was 86.36% ( P<0.001). Conclusion:The concentration of α-syn in CSF is related to the occurrence of POD, and it provides higher accuracy in predicting POD.

Objective:To evaluate the value of cerebrospinal fluid (CSF) nerve injury-related proteins levels in predicting postoperative delirium (POD) in patients.Methods:A total of 1 000 patients of both sexes, aged 40-90 yr, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, with Mini-Mental State Examination score>24 at 1 day before operation, undergoing elective knee/hip arthroplasty under spinal-epidural anesthesia , were enrolled in this study.Cubital venous blood samples were drawn before anesthesia for detection of the concentrations of plasma total cholesterol, high-density lipoprotein, low-density lipoprotein and triglyceride.CSF 2ml was extracted after successful spinal-epidural anesthesia puncture for measurement of concentrations of α-synuclein (α-syn), β-amyloid protein 1-40 (Aβ 1-40), Aβ 1-42, total-Tau (t-Tau), phosphorylated Tau (p-Tau), progranulin (PGRN) and soluble myeloid cell triggering receptor 2 (sTREM2) (by enzyme-linked immunosorbent assay). The Confusion Assessment Method was used at 1, 3 and 7 days after surgery to evaluate the occurrence of POD.The patients were divided into POD group and non-POD group according to whether POD occurred after operation.Logistic regression analysis was used to analyze the variables of which P values were less than 0.05 to analyze the risk factors for POD.The receiver operating characteristic (ROC) curve was drawn and area (AUC) under the curve was calculated to evaluate the accuracy of the related risk factors in predicting POD. Results:A total of 964 patients were enrolled in the study, and 108 patients were diagnosed with POD, and the incidence was 11.2%.The results of logistic regression analysis found that age and and increased α-syn in CSF concentration were risk factors for POD, and decreased PGRN in CSF concentration and Aβ 1-42/p-Tau in CSF were the protective factors for POD ( P<0.05). ROC curve analysis showed that α-syn (AUC 0.69, 95% confidence interval (CI) 0.634-0.748, sensitivity 57.41%, specificity 82.10%, Youden Index 0.3951), PGRN in CSF concentration (AUC 0.695, 95%CI 0.637-0.750, sensitivity 59.26%, specificity 80.86%, Youden Index 0.4012) and Aβ 1-42/p-Tau in CSF (AUC 0.635, 95%CI 0.574-0.692, sensitivity 93.52%, specificity 30.25%, Youden Index 0.2377) could predict the occurrence of POD. Conclusion:PGRN, α-syn concentration and Aβ 1-42/p-Tau in CSF can predict the occurrence of POD in patients.

Objective:To evaluate the relationship between preoperative cerebrospinal fluid/serum albumin ratio (Q-alb) and postoperative delirium (POD) in patients undergoing neuraxial anesthesia.Methods:The patients, aged 40-90 yr, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, underwent total knee/hip replacement under combined spinal-epidural block in our hospital from January 2018 to December 2020, were collected.After admission to the operating room, venous blood and cerebrospinal fluid samples were collected for determination of cerebrospinal fluid albumin, β-amyloid (Aβ) 1-42, Aβ 1-40, total tau protein (t-Tau), phosphorylated tau protein (p-Tau) and serum albumin levels (by enzyme-linked immunosorbent assay) and for calculation of Q-alb.When Q-alb was more than 10.2, the patient was considered to have blood-brain barrier disruption.Mini-Mental State Examination scale was used to evaluate the cognitive level on 1 day before surgery. The development of POD was evaluated using Confusion Assessment Method Chinese Reversion and Memorial Delirium Assessment Scale at 1-7 days after surgery.The patients were divided into POD group (P group) and non-POD (NP group) according to whether POD occurred.The receiver operating characteristic (ROC) curve was used to analyze the accuracy of Q-alb in predicting POD. Results:There were 49 cases in each group.Compared with group NP, concentrations of Aβ 1-42 and Aβ 1-40 were significantly decreased, concentrations of t-Tau and p-Tau albumin were increased, the ratio of Q-alb and blood-brain barrier disruption was increased in group P ( P<0.05). Before and after adjusting for confounding factors, Q-alb, cerebrospinal fluid Aβ 1-42, Aβ 1-40, t-Tau and p-Tau levels were risk factors for POD ( P<0.05). There was a positive linear regression relationship between Q-alb and levels of t-Tau and p-Tauin cerebrospinal fluid (t-Tau: β=0.587, P<0.001; p-Tau: β=0.427, P<0.001), and there was a negative linear regression relationship between Q-alb and levels of Aβ 1-42 and Aβ 1-40 in cerebrospinal fluid (Aβ 1-42: β=-0.762, P<0.001; Aβ 1-40: β=-0.531, P<0.001). There was no linear regression relationship between Q-alb and level of p-Tau in group P ( P=0.121). There was no linear regression relationship between Q-alb and level of Aβ 1-40 in group NP ( P=0.467). The results of ROC curve analysis showed that the area under the curve for Q-alb in predicting POD (95% confidence interval) was 0.827 (0.738-0.896). Conclusion:Preoperative higher Q-alb is the risk factor for POD in patients undergoing neuraxial anesthesia, and is more accurate in predicting POD.