Objective To investigate the effects of Echinococcus multilocularis infection on levels of memory T (Tm) cells and their subsets in lymph nodes of mice at different stages of infection, so as to provide new insights into immunotherapy for alveolarechinococcosis. MethodsTwenty-four C57BL/6J mice aged 6 to 9 weeks were randomly divided into the infection group and the control group, of 12 mice in each group. Mice in the infection group were administered with 3 000 E. multilocularis protoscoleces via portal venous injection, while animals in the control group were administered with an equal volume of physiological saline. Three mice from each group were sacrificed 4, 12 weeks and 24 weeks post-infection, and lymph nodes were sampled and stained with hematoxylin and eosin (HE) to investigate the histopathological changes of mouse lymph nodes in the infection group. The expression and localization of T lymphocyte surface markers CD3, CD4, and CD8 were observed in mouse lymph nodes using immunohistochemical staining. In addition, lymphocyte suspensions were prepared from mouse lymph nodes in both groups at different time points post-infection, and the levels of Tm cell subsets and their secreted cytokines were detected using flow cytometry. Results HE staining showed diffuse structural alterations in the subcapsular cortical and paracortical regions of mouse lymph nodes in the infection group 4 weeks post-infection with E. multilocularis. Immunohistochemical staining detected CD3, CD4 and CD8 expression in mouse lymph nodes in both groups. Flow cytometry revealed higher proportions of CD4⁺ Tm cells [(55.3 ± 4.8)% vs. (38.8 ± 6.1)%; t = -4.259, P < 0.05] and CD4⁺ tissue-resident Tm (Trm) cells [(57.7 ± 3.7)% vs. (34.1 ± 11.2)%; t = -3.990, P < 0.05] in mouse lymph nodes in the infection group than in the control group 4 weeks post-infection, and higher proportions of CD4⁺ Tm cells [(34.6 ± 3.2)% vs. (23.3 ± 7.5)%; t = -2.764, P < 0.05] and CD4⁺ Trm cells [(44.0 ± 1.9)% vs. (31.2 ± 1.5)%; t = -4.039, P < 0.05] in mouse lymph nodes in the infection group than in the control group 24 weeks post-infection. The proportions of CD8⁺ Tm cells were higher in the infection group than in the control group 4 weeks [(56.8 ± 2.7)% vs. (43.9 ± 5.2)%; t = -4.416, P < 0.01] and 12 weeks post-infection [(25.4 ± 2.7)% vs. (12.0 ± 2.6)%; t = -2.552, P < 0.05], while the proportions of tumor necrosis factor (TNF)-α⁺ CD4⁺ T cells [(15.7 ± 5.0)% vs. (49.4 ± 6.4)%; t = 7.150, P < 0.01], TNF-α⁺CD8⁺ T cells [(20.7 ± 5.5)% vs. (57.5 ± 8.4)%; t = -6.694, P < 0.01], and TNF-α⁺ CD8⁺ Tm cells [7.0% (1.0%) vs. 31.0% (11.0%); Z = -2.236, P < 0.05] were lower in the infection group than in the control group 24 weeks post-infection. Conclusions Tm cells levels are consistently increased in lymph nodes of mice at different stages of E. multilocularis infection, with Trm cells as the predominantly elevated subset. The impaired capacity of CD8⁺ Tm cells to secrete the effector molecule TNF-α in mouse lymph nodes at the late-stage infection may facilitate chronic parasitism of E. multilocularis.

Objective To investigate the effect of tritiated water on the immune system of zebrafish and its potential molecular mechanism. Methods Zebrafish embryos (2.5 to 3 hours post-fertilization [hpf]) were exposed to 3.7 × 10⁴ Bq/mL tritiated water (tritiated water group), and those exposed to E3 culture medium were used as the control group. The mortality rate, hatching rate, deformity rate, heart rate, body length, yolk sac area, neutrophil count in the tail, immune-related gene expression, and immune-related protein expression of zebrafish in the two groups were determined. Then transcriptome technology was used to further analyze the possible mechanism of tritiated water affecting the immune system of zebrafish. Results Compared with the control group, zebrafish at 72 hpf in the tritiated water group had no significant changes in the mortality rate, hatching rate, deformity rate, body length, and yolk sac area((t = 0.9045, 0.5000, 1.0000, 0.7238, 0.0337, P = 0.4169, 0.6433, 0.3739, 0.4785, 0.9735), but had significantly increased heart rate(t = 4.575，P = 0.002). At 4 days post-fertilization (dpf), the neutrophil count in the tail of zebrafish in the tritiated water group was significantly increased(t = 2.563，P = 0.0196), the mRNA expression of TNF-α was significantly decreased(t = 2.891, P = 0.045), the protein expression of nuclear factor-kappa B (NF-κB) was significantly increased(t = 3.848, P = 0.018), and the protein expression of NLRP3 was significantly decreased(t = 14.98, P = 0.001). At 7 dpf, the neutrophil count in the tail and the protein expression levels of NF-κB, NLRP3, and interleukin-1β were significantly decreased(t = 3.772, 7.048, 15.620, 4.423, P = 0.014, 0.002, 0.0001, 0.012). Transcriptome sequencing revealed that differentially expressed genes were mainly enriched in the “neutrophil activation” and “platelet activation pathways” at 4 dpf and in the “neutrophil apoptosis”, “ferroptosis”, and “necroptosis” pathways at 7 dpf. Conclusion Tritiated water exposure induces a temporally dynamic immune response in zebrafish, potentially affecting immune homeostasis by regulating neutrophil activation and apoptosis, as well as the expression of NF-κB and NLRP3.

ObjectiveTo analyze the distribution, drug resistance characteristics, and changing trends of Acinetobacter baumannii (AB) isolated from environmental surfaces and healthcare workers’ hands in a grade Ⅱ level A general hospital in Xuhui District of Shanghai from 2018 to 2023, and to provide reference for infection control in the hospital. MethodsEnvironmental samples were collected quarterly from critical surfaces and healthcare workers’ hands in the intensive care unit (ICU), geriatrics, and respiratory departments from 2018 to 2023. Clinical isolates were obtained from all patients with AB infections in ICU, geriatrics, respiratory department, rehabilitation department, infectious diseases department, emergency department, cardiology department, and orthopedics of the hospital from 2018 to 2023. Retrospective analyses were performed on AB detection rates, strain origins, resistance rates to commonly used antimicrobial agents, and resistance gene features, comparing the antimicrobial resistance between clinically isolated strains and environmentally isolated strains. ResultsFrom 2018 to 2023, a total of 1 416 samples were collected from the hospital and a total of 272 strains of AB were detected, with a positive detection rate of 19.21%. The detection rate gradually decreased year-on-year (χ²_trend=45.290, P<0.001). The majority of samples originated from patient-contacted items (34.56%, 94/272), followed by shared items (26.84%, 73/272) and healthcare worker-contacted items (15.07%, 41/272). From 2018 to 2023, the resistance rate of AB on environmental surfaces and healthcare workers’ hands to commonly tested antibiotics in the hospital ranged from 10% to 40%. The resistance rates to cefotaxime (42.52%) and piperacillin (38.58%) were relative high, while the resistance to polymyxin E (1.57%), polymyxin B (2.36%), and doxycycline (3.94%) maintained low. The annual fluctuations in resistance to cefotaxime, piperacillin, ceftriaxone, tobramycin, doxycycline, minocycline and cotrimoxazole were statistically significant (all P<0.05). There were statistically significant differences in the resistance of clinical and environmental isolates to ampicillin/sulbactam, cefepime, ceftazidime, subamphetamine, meropenem, piperacillin, aztreonam, gentamicin, tobramycin, minocycline, ciprofloxacin, levofloxacin, and cotrimoxazole in the hospital from 2018 to 2023 (all P<0.05). The resistance rate of clinical isolates was generally high, especially to β-lactam and quinolone drugs, which were mostly above 80% [such as cefepime (93.86%), cefotaxime (97.37%), imipenem (98.25%), and ciprofloxacin (99.12%)]. The resistance rate of environmental isolated strains to similar antibiotics was relatively lower, mostly concentrated at 10%‒30%. The whole-genome sequencing of 34 carbapenem-resistant Acinetobacter baumannii (CRAB) strains isolated from the hospital environment in 2023 revealed that the main resistance mechanism was overexpression of efflux pumps (51.97%), followed by changes in target sites (32.46%). Among the 34 CRAB strains, carbapenem resistance genes OXA-23 and OXA-51 were detected in 6 strains (17.65%), while genes such as KPC, IMP, VIM, and SIM were not detected. ConclusionFrom 2018 to 2023, AB in the hospital environment exhibited high resistance rates to certain antimicrobial agents and carried multiple resistance genes, indicating a potential transmission risk. It is necessary to further strengthen bacterial resistance monitoring and hospital infection control, and use antibiotics reasonably.

Objective To investigate the effect of LAG-3 deficiency (LAG3^-/-) on natural killer (NK) cell function and hepatic fibrosis in mice infected with Echinococcus multilocularis. Methods C57BL/6 mice, each weighing (20 ± 2) g, were divided into the LAG3^-/- and wild type (WT) groups, and each mouse in both groups was inoculated with 3 000 E. multilocularis protoscoleces via the hepatic portal vein. Mouse liver and spleen specimens were collected 12 weeks post-infection, sectioned and stained with sirius red, and the hepatic lesions and fibrosis were observed. Mouse hepatic and splenic lymphocytes were isolated, and flow cytometry was performed to detect the proportions of hepatic and splenic NK cells, the expression of CD44, CD25 and CD69 molecules on NK cell surface, and the secretion of interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), interleukin (IL)-4, IL-10 and IL-17A. Results Sirius red staining showed widening of inflammatory cell bands and hyperplasia of fibrotic connective tissues around mouse hepatic lesions, as well as increased deposition of collagen fibers in the LAG3-/-group relative to the WT group. Flow cytometry revealed lower proportions of mouse hepatic (6.29% ± 1.06% vs. 11.91% ± 1.85%, P < 0.000 1) and splenic NK cells (4.44% ± 1.22% vs. 5.85% ± 1.10%, P > 0.05) in the LAG3^-/- group than in the WT group, and the mean fluorescence intensity of CD44 was higher on the surface of mouse hepatic NK cells in the LAG3^-/- group than in the WT group (t = −3.234, P < 0.01), while no significant differences were found in the mean fluorescence intensity of CD25 or CD69 on the surface of mouse hepaticNK cells between the LAG3^-/- and WT groups (both P values > 0.05). There were significant differences between the LAG3^-/- and WT groups in terms of the percentages of IFN-γ (t = −0.723, P > 0.05), TNF-α (t = −0.659, P > 0.05), IL-4 (t = −0.263, P > 0.05), IL-10 (t = −0.455, P > 0.05) or IL-17A secreted by mouse hepatic NK cells (t = 0.091, P > 0.05), and the percentage of IFN-γ secreted by mouse splenic NK cells was higher in the LAG3^-/- group than in the WT group (58.40% ± 1.64% vs. 50.40% ± 4.13%; t = −4.042, P < 0.01); however, there were no significant differences between the two groups in terms of the proportions of TNF-α (t = −1.902, P > 0.05), IL-4 (t = −1.333, P > 0.05), IL-10 (t = −1.356, P > 0.05) or IL-17A secreted by mouse splenic NK cells (t = 0.529, P > 0.05). Conclusions During the course of E. multilocularis infections, LAG3^-/- promotes high-level secretion of IFN-γ by splenic NK cells, which may participate in the reversal the immune function of NK cells, resulting in aggravation of hepatic fibrosis.

Objective:To study the localization value of stereoelectroencephalography (SEEG) in refractory epilepsy associated with tuberous sclerosis complex (TSC), and to analyze the prognosis related factors.Methods:A case control study.Data of 66 patients with TSC-related refractory epilepsy who underwent SEEG placement at the Epilepsy Center of Tsinghua University Yu-Quan Hospital from January 2014 to March 2023 were retrospectively analyzed, including medical history, seizure semiology, scalp electroencephalogram (EEG), imaging, and SEEG findings.Patients were divided into a seizure-free group and a seizure group according to the seizures after surgery. t/ χ2 test was used to compare the relationship between age of onset, age of surgery, duration of epilepsy, genetic findings, seizure symptoms, scalp EEG characteristics, SEEG placement protocol and prognosis. Results:A total of 66 patients met the enrollment criteria, of whom 55 patients were seizure-free after surgery (seizure-free group), and 11 patients still had seizures (seizure group).Among the 66 patients, there were 46 males and 20 females; the age of onset was (21.72±30.01) months; the age of SEEG surgery was (6.79±5.12) years; the duration of epilepsy was (4.98±4.11) years.SEEG results showed that seizures started with a single tuber in 43 patients (64.4%), seizures started with 2 or more tubers in one hemisphere in 16 patients, and 7 patients had bilateral multi-tubers or the pathogenic tuber(s) could not be identified.The following factors were found to be associated with prognosis: early age of surgery ( t=-3.463, P<0.01), ictal scalp EEG with a definite lateralization ( χ2=7.876, P<0.05), and the concordance of interictal and ictal EEG ( χ2=6.821, P<0.05).The age of onset, duration of epilepsy, seizure type and symptom, genetic findings, and SEEG placement protocol were not significantly correlated with prognosis.The results of SEEG identified that onset with a single tuber revealed a better postoperative outcome. Conclusions:This study confirms the safety and efficacy of SEEG in TSC-related refractory epilepsy, and also finds that ictal EEG is uniquely valuable in guiding SEEG placement for TSC-related refractory epilepsy, which can help us better select patients with TSC-related refractory epilepsy who are suitable for SEEG placement.

Objective To analyze the relationship between expression levels of long non-coding RNA(LncRNA)LINC00342 and microRNA-203a-3p(miR-203a-3p)in colorectal cancer tissue and the prognosis of patients within 5 years after surgery.Methods Samples of colorectal cancer tissue and paracancer tissue of 133 patients with colorectal cancer were collected.Expression levels of LncRNA LINC00342 and miR-203a-3p were detected by fluorescence quantitative PCR.Survival and death were recorded after 5 years of follow-up.The LncRNA LINC00342 and miR-203a-3p expressions and clinicopathological parameters in patients under different conditions were compared.The correlation between expressions of LncRNA LINC00342 and miR-203a-3p in colorectal cancer tissue was analyzed,and their predictive value for the prognosis was also analyzed.Results The expression level of LncRNA LINC00342 was higher in colorectal cancer tissue than that in paracancer tissue,and the expression level of miR-203a-3p was lower in colorectal cancer tissue than that in paracancer tissue(P＜0.05).The expression levels of LncRNA LINC00342 and miR-203a-3p in colorectal cancer tissue were negatively correlated(P＜0.05).The proportion of patients with low tumor differentiation,TNM stage Ⅲ and lymph node metastasis were higher in the LncRNA LINC00342 high expression group and the miR-203a-3p low expression group than those in the LncRNA LINC00342 low expression group and the miR-203a-3p high expression group,respectively(P＜0.05).The 5-year overall survival rate after operation was lower in the LncRNA LINC00342 high expression group and the miR-203a-3p low expression group(P＜0.05).The proportion of patients with low tumor differentiation,TNM stage Ⅲ,lymph node metastasis,and higher expression level of LncRNA LINC00342 were higher in colorectal cancer tissue of the death group than those in the survival group,and the expression level of miR-203a-3p was lower than that in the survival group(P＜0.05).Low tumor differentiation,TNM stage Ⅲ,lymph node metastasis,high expression of LncRNA LINC00342 and low expression of miR-203a-3p were independent risk factors for death within 5 years after colorectal cancer surgery(P＜0.05).The combined predictive value of LncRNA LINC00342 and miR-203a-3p for prognosis was greater than that of individual prediction.Conclusion LncRNA LINC00342 is higher expressed and miR-203a-3p is lower expressed in colorectal cancer tissue,and the combined detection of the two is expected to become a clinical evaluation indicator for predicting postoperative survival.

Background::Transplant renal artery stenosis (TRAS) is a vascular complication after kidney transplantation associated with poor outcomes. This study aimed to analyze the efficacy and safety of low-dose aspirin for preventing TRAS.Methods::After kidney transplantation, patients were enrolled from January 2018 to December 2020 in Henan Provincial People’s Hospital. A total of 351 enrolled recipients were randomized to an aspirin group with low-dose intake of aspirin in addition to standard treatment ( n = 178), or a control group with only standard treatment ( n = 173). The patients was initially diagnosed as TRAS (id-TRAS) by Doppler ultrasound, and confirmed cases were diagnosed by DSA (c-TRAS). Results::In the aspirin and control groups, 15.7% (28/178) and 22.0% (38/173) of the recipients developed id-TRAS, respectively, with no statistical difference. However, for c-TRAS, the difference of incidence and cumulative incidence was statistically significant. The incidence of c-TRAS was lower in the aspirin group compared with the control group (2.8% [5/178] vs. 11.6% [20/173], P = 0.001). Kaplan–Meier estimates and Cox regression model identified the cumulative incidence and hazard ratio (HR) of TRAS over time in two groups, showing that recipients treated with aspirin had a significantly lower risk of c-TRAS than those who were not treated (log-rank P = 0.001, HR = 0.23, 95% confidence interval [CI]: 0.09–0.62). The levels of platelet aggregation rate ( P < 0.001), cholesterol ( P = 0.028), and low-density lipoprotein cholesterol ( P = 0.003) in the aspirin group were decreased compared with the control group in the third-month post-transplantation. For the incidence of adverse events, there was no statistical difference. Conclusion::Clinical application of low-dose aspirin after renal transplant could prevent the development of TRAS with no significant increase in adverse effects.Trial Registration::Clinicaltrials.gov, NCT04260828.

Objective:To explore the safety and reliability of retrosigmoid approach BONEBRIDGE implantation in patients with auricle reconstruction using skin expansion flap.Methods:A retrospective analysis was conducted on 43 congenital aural atresia cases (43 ears) who underwent BONEBRIDGE implantation from September 2019 to January 2023 in Beijing Tongren Hospital. 30 males and 13 females were included in this work. The implantation age was 9-36 years old (median age=10 y/o). All cases underwent auricle reconstruction surgery using the posterior ear flap expansion method, with 36 cases using the single expanded postauricular flap method and 7 cases using two-flap method. BONEBRIDGE implant surgery was performed during the third stage of auricle reconstruction or after all stages. The hearing improvements were evaluated by comparing the changes in pure tone hearing threshold and speech recognition rate of patients before and after BONEBRIDGE implantation. Routine follow-up was conducted to observe the hearing results and complications. SPSS 14.0 software was applied for data statistical analysis.Results:All 43 patients healed well and had no surgical complications when discharge. The average bone conduction hearing threshold after surgery was (8.2±6.6) dBHL, and there was no statistically significant difference compared to the preoperative [(8.1±5.7) dBHL] ( P=0.95). After surgery, the threshold of hearing assistance with power on was significantly lower than that without hearing assistance [(32.8±4.6) dBHL vs (60.5±5.5) dBHL], and the difference was statistically significant ( P<0.001). The speech recognition rate of monosyllable words, disyllabic words and short sentences in quiet environment increased to 72%, 84%, and 98% respectively. The differences were statistically significant ( P<0.001). The speech recognition rate of monosyllabic words, disyllabic words, and short sentences in noise environment was significantly increased by 70%, 80%, and 92% respectively ( P<0.001). After a follow-up of 4 to 47 months (median=24 months), the hearing results were stable and the aesthetic outcomes were satisfying. One patient had delayed hematoma around coil of the implant. After aspiration and compressed dressing for one week, hematoma was not recurrent. Conclusion:For patients after auricle reconstruction using expanded postauricular flap, the preference of retrosigmoid approach is a good choice in terms of safety and reliability of operation, as well as aesthetic appearance.

Objective:To record stereoelectroencephalography (SEEG) data and to induce cortical electrical stimulation in children with tuberous sclerosis complex (TSC), thus exploring the epileptogenicity of different types of cortical tubers.Methods:The SEEG recording and cortical electrical stimulation data of 50 children with TSC who underwent preoperative evaluation for drug-resistant epilepsy at Epilepsy Center, Tsinghua University Yuquan Hospital from November 2016 to September 2022 were retrospectively analyzed, involving 27 boys and 23 girls with the age of (5.5±3.4) years.According to the results of 3.0T magnetic resonance imaging (3T-MRI) and computed tomography(CT), cortical tubers were classified.The incidences of electroclinical seizures, electrical seizures and seizures induced by cortical electrical stimulation in different types of tubers recorded by SEEG were analyzed, and the differences in the proportion of the above seizures among different types of tubers were compared using the Fisher′ s exact test. Results:A total of 303 cortical tubers were explored using SEEG in 50 patients.The tubers were divided into 6 types, including Type A, B, C, D and E, and focal cortical dysplasia like (FCD-like) type, among which Type E was for the first time proposed in the world.Among these explored tubers, 7 tubers had electrical seizures, and 57 tubers had electroclinical seizures.A total of 64 tubers (21.1%) were epileptogenic.The incidence of epileptogenic tubers in Type A-E and FCD-like type were 3.6%, 1.4%, 19.0%, 77.8%, 77.5%, and 90.0%, respectively. Fisher′ s exact test and Bonferroni correction were performed for pairwise comparisons( P<0.003). There was no significant difference in the incidence of epileptogenic tubes among Type A, B and C. There was significant difference in the incidence of epileptogenic tubes between Type A-C with Type D, Type E and FCD-like type, respectively.There was no significant difference in the incidence of epileptogenic tubes between Type D, Type E and FCD-like like.Electrical stimulation-induced seizures occurred in 36 cortical tubers (11.9%). The positive rate of electrical stimulation seizures in Type A-E and FCD-like type were 0.7%, 1.4%, 4.8%, 44.4%, 45.0%, and 70.0%, respectively.There was significant difference in the positive rate of electrical stimulation seizures between Type A-B and Type D, Type E and FCD-like type, respectively, so as that between Type C versus Type E and FCD-like type.No significant difference in the positive rate of electrical stimulation seizures was found between other pairwise comparisons. Conclusions:This study proposed a new classification of cortical tubers in TSC patients, and Type E is proposed for the first time in the world.SEEG records confirmed great differences in epileptogenicity indifferent types of cortical tubers.Type D, Type E and FCD-like type have higher epileptogenicity, which is of great value for the preoperative evaluation of TSC epilepsy surgery and the placement strategy of SEEG electrodes.

Objective:To evaluate the prevalence and related risk factors of new onset diabetes after chronic pancreatitis by meta-analysis.Methods:Chronic pancreatitis, diabetes, post pancreatitis diabetes, type 2 diabetes, type 3c diabetes, endocrine dysfunction, chronic pancreatitis, diabetes mellitus, post pancreatitis diabetes mellitus, endocrine efficiency, risk factors were used as keywords, and the network database such as the CNKI database, Wanfang, Weipu, Chinese Medical Journal Full Text, PubMed, Embase, Cochrane Library, Web of Science, and so on from the database establishment to January 2023 were searched. The prospective and retrospective cohort studies on new diabetes after chronic pancreatitis published were searched and retrieved, and the papers were screened and the quality were evaluated according to preset inclusion and exclusion criteria; and the important data were extracted. Review Manager 5.4 was used for meta-analysis.Results:22 papers were finally included, including 13 785 patients with chronic pancreatitis, of which 4 233 were patients with new onset diabetes. Meta-analysis showed that the incidence of new diabetes after chronic pancreatitis was 29% ( RD=0.29, 95% CI 26%-32%, P<0.0001), which increased and tended to be stable along with the disease course. Alcohol drinking, smoking, alcoholic chronic pancreatitis, pancreatic calcification, biliary stricture, male, conservative treatment, pancreatic cyst and older onset age were considered as risk factors for new diabetes after chronic pancreatitis, and endoscopic treatment was considered as protective factors. Conclusions:The incidence of new diabetes after the diagnosis of chronic pancreatitis is relatively high. Clinically, we can identify high-risk groups exposed to risk factors, and early intervention can reduce the incidence rate of new diabetes after chronic pancreatitis and improve the prognosis of patients.