This study aims to determine whether 4 weeks of adjunctive minocycline can improve cognitive and neurological symptoms in patients with autoimmune encephalitis (AE). In this randomized, open-label, blinded-endpoint trial, participants were randomized (1:1) to receive first-line immunotherapy with or without minocycline (100 mg twice daily for 4 weeks). Clinical outcomes included changes in the Montreal Cognitive Assessment (MoCA), Mini-Mental State Examination (MMSE), Hamilton Anxiety Rating Scale (HAMA), Hamilton Depression Rating Scale (HAMD), modified Rankin Scale (mRS), and Clinical Assessment Scale in Autoimmune Encephalitis (CASE). The exploratory outcome was the change in Soluble Triggering Receptor Expressed on Myeloid cells 2 (sTREM2), a biomarker of microglial activation. Follow-up assessments were conducted at weeks 4, 12, 24, and 48. Sixty patients with AE were enrolled. For the primary outcome, the difference in mean change in MoCA scores between the groups was 1.05 (P = 0.46) from baseline to week 12. However, at week 4, the minocycline group demonstrated significantly improvements in MoCA (difference, 3.85; 95

OBJECTIVES:We aimed to explore the impact of glymphatic function in patients diagnosed with autoimmune encephalitis (AE). METHODS:In this prospective longitudinal study, patients were recruited from Xijing Hospital between June 2020 and January 2024. Glymphatic function was evaluated using diffusion tensor imaging analysis along the perivascular space (DTI-ALPS). Cognitive impairment was defined as a Montreal Cognitive Assessment (MoCA) score below 26 at the 12-month follow-up. RESULTS:A total of 115 individuals were enrolled, including 85 patients with AE and 30 age- and sex-matched healthy controls (HCs). After correcting for age and sex, patients with AE had a significantly lower baseline ALPS index compared to HCs (1.173, 95 % CI [1.135, 1.210] vs. 1.456, 95 % CI [1.371, 1.541]; P < 0.001). The baseline ALPS index was correlated with cognitive performance, including a positive correlation with the Mini-Mental State Examination (MMSE) score (r = 0.568, P < 0.001) and a positive correlation with the MoCA score (r = 0.645, P < 0.001). In the longitudinal study, the ALPS index gradually increased over the follow-up period (P < 0.001), and a low level of the baseline ALPS index was associated with a higher risk of long-term cognitive impairment (HR [95 % CI] = 1.70 [1.12-2.58], P = 0.013). CONCLUSION:The glymphatic system is impaired in AE patients. A decreased DTI-ALPS index is associated with a decline in cognitive performance. Additionally, a low baseline ALPS index may predict an increased risk of long-term cognitive impairment in AE patients.

BACKGROUND AND OBJECTIVE:Autoimmune encephalitis (AE) is an immune-mediated disease. Some patients experience persistent cognitive deficits despite receiving immunotherapy. We aimed to develop a prediction model for long-term cognitive outcomes in patients with AE. METHOD:In this multicenter cohort study, a total of 341 patients with AE were enrolled from February 2014 to July 2023. Cognitive impairment was identified using the telephone Mini-Mental State Examination (t-MMSE). Six machine learning (ML) algorithms were used to assess the risk of developing cognitive impairment. RESULTS:The median age of the patients with AE was 30.0 years (23.0-48.25), and 48.90 % (129/264) were female in the training cohort.77 (29.2 %) patients were identified with cognitive impairment after a median follow-up of 49 months. Among 16 features, the following six features were finally selected to develop the model: Cognitive Reserve Questionnaire (CRQ), Clinical Assessment Scale for Autoimmune Encephalitis (CASE), status epilepticus (SE), age, MRI abnormalities, and delayed immunotherapy. Compared to other ML models, the random forest (RF) model demonstrated superior performance with an AUC of 0.90. The accuracy, sensitivity, and specificity in the testing cohort were 0.87, 0.79, and 0.90, respectively. CONCLUSION:The RF model based on CRQ, CASE scores, SE, age, MRI abnormalities and delayed immunotherapy demonstrates superior predictive performance and shows promise in predicting the risk of long-term cognitive outcomes in patients with AE in clinical settings.

Aim To investigate the prevalence and clinical features of kidney injury in patients with autoimmune encephalitis (AE). Methods Kidney injury was suspected in kidney-involving group due to persistent abnormal in urinary protein and serum albumin. Data on demographics and clinical features were compared between kidney-involving group and kidney-sparing group (patients without kidney injury) using Wilcoxon rank-sum test or chi-square test. Renal biopsy was conducted to identify the type of kidney injury. Results Approximate 30 % (32 of 108) patients with AE were suspicious of kidney injury. Nine patients further tested 24 h urine total protein, and seven of them had an elevated urine protein higher than 150 mg. The predominantly patterns of kidney injury were elevated urine protein, decreased serum albumin and normal kidney function. Compared to kidney-sparing group, the spectrum of AE antibodies in kidney-involving group was different, manifested as less anti-N-methyl-d-aspartate receptor antibodies (50 % vs. 72.4 %, p = 0.025) and more anti-contactin-associated protein like 2 antibodies (18.8 % vs. 1.3 %, p = 0.003). Definite pathological changes indicative of IgA nephropathy and membranous nephropathy in renal biopsy of two cases provided evidence of autoimmune attacks. Discussion Kidney injury occurred in considerable proportion of patients with AE. An in-depth screening for nephropathy could be essential for AE.

Objective:To investigate the clinical and electrophysiological characteristics of patients with sudden unexpected death of epilepsy (SUDEP).Methods:Using "epilepsy" as the keyword, the relevant cases entered from October 2011 to March 2012 were searched in the database of the Electroencephalogram (EEG) Monitoring Center, Xijing Hospital, the Air Force Military Medical University. Telephone follow-up was conducted for all confirmed epilepsy patients, and for the death cases confirmed by telephone follow-up, the patients identified as consistent with SUDEP diagnosis were included in this study based on their past medical history, clinical data, death details, etc, and their clinical and neuroelectrophysiological characteristics were summarized and analyzed.Results:Among the 1 232 patients who underwent 24-hour video-EEG monitoring during the study period, 354 patients were successfully followed up by telephone interview, of whom 17 patients were died (4.8%), 12 individuals met the diagnosis of SUDEP (7 men, 5 women). The duration of the disease in 9 patients exceeded 10 years. Eight cases presented with focal-bilateral tonic clonic seizures. Nine patients were treated with anti-seizure drug monotherapy. All the 24-hour video EEG of 12 patients were abnormal. There were 8 occasions when the EEG occipital α background rhythm slowed down compared with the standard frequency of peers or was dominated by slow waves. Interictal epileptic discharge (IED) located in temporal lobe were found in 12 EEG records, of which 9 EEG records were found with frontal IED. One of the 12 cases received 24-hour video EEG twice within 6 years, and his EEG background rhythm was significantly slower and the IED region was expanded compared with the first EEG record. At the third year after reexamination of EEG, SUDEP developed in this patient.Conclusions:SUDEP patients have a long course of disease and bilateral tonic-clonic seizure. The interictal EEG shows occipital slow α activity and temporofrontal epileptiform discharges, which may increase the risk of SUDEP.

Cerebrovascular events, especially ischemic stroke, are common complications of essential thrombocythemia (ET). Compared to JAK2V617 F mutation, CALR mutation is considered as a lower risk factor of thrombosis in ET. Until now stroke in ET with CALR mutation has rarely been reported. We retrospectively investigated patients diagnosed with stroke and ET in Xijing hospital of Air Force Medical University, from 2015 to 2021. Clinical characteristics (including medical history, physical and auxiliary examination and prognosis) were recorded and associated literature was reviewed. Among the 19 patients diagnosed with both stroke and ET we retrieved, two cases were positive for CALR mutation. In case 1, a 71-year-old man developed the first ischemic event under the treatment of anagrelide, followed by a hemorrhagic stroke after receiving aspirin and clopidogrel for 4 months. Ischemic stroke reccurred and the neurological function deteriorated progressively. In case 2, a 44-year-old man presented with hypoxic-ischemic encephalopathy due to serious myocardial infarction and subsequent brain imaging indicated three times of ischemic stroke events. The patient gradually got improved through cytoreductive and antiplatelet therapy and rehabilitation. Literature review showed that cerebrovascular event is the most serious neurological complication of ET and may be the presenting symptom. Most of reported cases with ET accompanied by stroke were positive for JAK2 V617 F mutation, but with rare CALR mutation. ET with CALR mutation can cause both hemorrhagic and ischemic stroke. Identification of such rare causes of stroke is of great importance to provide precise and individualized prevention and therapy.

Background Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a type of autoimmune encephalitis. The underlying mechanism(s) remain largely unknown. Recent evidence has indicated that the gut microbiome may be involved in neurological immune diseases via the "gut-brain axis". This study aimed to explore the possible relationship between anti-NMDAR encephalitis and the gut microbiome. Methods Fecal specimens were collected from 10 patients with anti-NMDAR encephalitis and 10 healthy volunteers. The microbiome analysis was based on Illumina sequencing of the V3-V4 hypervariable region of the 16S rRNA gene. The alpha, beta, and taxonomic diversity analyses were mainly based on the QIIME2 pipeline. Results There were no statistical differences in epidemiology, medication, and clinical characteristics (except for those related to anti-NMDAR encephalitis) between the two groups. ASV analysis showed that Prevotella was significantly increased, while Bacteroides was reduced in the gut microbiota of the patients, compared with the controls. Alpha diversity results showed a decrease in diversity in the patients compared with the healthy controls, analyzed by the Shannon diversity, Simpson diversity, and Pielou_E uniformity based on the Kruskal-Wallis test (P = 0.0342, 0.0040, and 0.0002, respectively). Beta diversity analysis showed that the abundance and composition of the gut microbiota was significantly different between the two groups, analyzed by weighted and unweighted UniFrac distance (P = 0.005 and 0.001, respectively). Conclusions The abundance and evenness of bacterial distribution were significantly lower and jeopardized in patients with anti-NMDAR encephalitis than in healthy controls. Thus, our findings suggest that gut microbiome composition changes might be associated with the anti-NMDAR encephalitis. It could be a causal agent, or a consequence.

目的 研究和开发针对非人类免疫缺陷病毒(HIV)人群中结核性脑膜炎和隐球菌性脑膜炎鉴别诊断的列线图模型,并对模型进行验证.方法 回顾性收集西京医院神经内科收治的292名结核性脑膜和隐球菌性脑膜炎患者的临床资料,根据入院时间将240名患者纳入训练组,52名患者纳入验证组.对训练组患者进行单因素和多因素分析,筛选鉴别两种脑膜炎的差异因子.利用R软件构建鉴别诊断列线图模型,并对模型进行验证.在训练组和验证组中绘制ROC曲线和校准曲线对模型进行评价.结果 单因素和多因素分析后发现,患者年龄、发热、自身免疫性疾病、脑脊液初压、脑脊液白细胞计数、脑脊液糖含量是鉴别结核性脑膜炎与隐球菌性脑膜炎的差异因子(P＜0.05).模型灵敏度为80％,特异度为76.47％.列线图模型ROC曲线下面积在训练组和验证组中分别为0.853和0.897,模型区分度和校准度较好.结论 在非HIV人群中结核性脑膜炎与隐球菌性脑膜炎的鉴别诊断列线图模型,对于基层医院进行脑膜炎的早期诊断和治疗具有一定的临床应用价值.

Cerebrospinal fluid (CSF) circulating in the human central nervous system has long been considered aseptic in healthy individuals, because normally, the blood-brain barrier can protect against microbial invasions. However, this dogma has been called into question by several reports that microbes were identified in human brains, raising the question of whether there is a microbial community in the CSF of healthy individuals without neurological diseases. Here, we collected CSF samples and other samples, including one-to-one matched oral and skin swab samples (positive controls), from 23 pregnant women aged between 23 and 40 years. Normal saline samples (negative controls), sterile swabs, and extraction buffer samples (contamination controls) were also collected. Twelve of the CSF specimens were also used to evaluate the physiological activities of detected microbes. Metagenomic and metatranscriptomic sequencing was performed in these 116 specimens. A total of 620 nonredundant microbes were detected, which were dominated by bacteria (74.6%) and viruses (24.2%), while in CSF samples, metagenomic sequencing found only 26 nonredundant microbes, including one eukaryote, four bacteria, and 21 viruses (mostly bacteriophages). The beta diversity of microbes compared between CSF metagenomic samples and other types of samples (except negative controls) was significantly different from that of the CSF self-comparison. In addition, there was no active or viable microbe in the matched metagenomic and metatranscriptomic sequencing of CSF specimens after subtracting those also found in normal saline, DNA extraction buffer, and skin swab specimens. In conclusion, our results showed no strong evidence of a colonized microbial community present in the CSF of healthy individuals. IMPORTANCE The microbiome is prevalent throughout human bodies, with profound health implications. However, it remains unclear whether it is present and active in human CSF, which has been long considered aseptic due to the blood-brain barrier. Here, we applied unbiased metagenomic and metatranscriptomic sequencing to detect the presence of a microbiome in CSF collected from 23 pregnant women with matched controls. Analysis of 116 specimens found no strong evidence to support the presence of a colonized microbiome in CSF. Our findings will strengthen our understanding of the internal environment of the CSF in healthy people, which has strong implications for human health, especially for neurological infections and disorders, and will help further disease diagnostics, prevention, and therapeutics in clinical settings.

We found that NGS technology has great significance for the diagnosis of brucellosis in non-epidemic area and can rule out other infections. The number of Brucella gene fragments detected by NGS may be affected by clinical resistance to Brucella treatment and Brucella infection. The sensitivity of SAT to detect Brucella may is limited.

中枢神经系统感染性疾病是一种凶险的神经科疾病,致死、致残率高,早诊断、早治疗是临床亟待解决的关键问题.随着分子诊断技术的不断进步,宏基因组二代测序技术(mNGS)在感染领域体现的突出作用而倍受关注.目前,研究报道mNGS技术对罕见病、特殊病原体的检测具有优势,越来越多的证据支持mNGS技术可以作为一种中枢神经系统感染性疾病筛查的重要手段,但是mNGS技术仍然不能完全取代传统实验室检测方法.文中就mNGS技术在中枢神经系统感染性疾病诊断中的应用、责任病原体的确定以及在中枢神经系统感染领域的研究方向进行综述,旨在帮助临床医生了解mNGS技术,对中枢神经系统感染性疾病进行更精准的诊断和治疗.

Paraneoplastic cerebellar degeneration (PCD) can occur in patients with underlying cancer, such as small cell lung cancer (SCLC). Anti-CV2/CRMP5 antibodies are well-established biomarkers of PCD associated with SCLC, but cannot be detected in most situations. Recently, next-generation sequencing has been a promising technology to discover cancer-driven mutations, which provide an alternative strategy to accomplish ultra-early diagnosis of those patients. Here, we report the case of a 75-year-old man diagnosed with SCLC, who primarily presented with anti-CV2/CRMP5 antibodies positive PCD. Eight high-frequency gene mutations (TSC2, DNMT1, CIC, FGF6, NSD1, TSHR, CRLF2, and EPPK1) were detected 7 months before diagnosis with no abnormalities of imaging or cerebrospinal fluid examination found initially. Therefore, this case suggests the possibility of detecting certain somatic mutations for the ultra-early diagnosis of patients presenting with PCD associated with SCLC.

目的 比较隐球菌性脑膜炎(CM)不同检测方法 的诊断价值.方法 选取2012年1月—2018年11月西京医院神经内科脑脊液细胞学检查室CM患者42例的脑脊液样本47份作为病例组,非CM脑脊液样本40份作为对照组,分别使用脑脊液墨汁染色、荚膜抗原胶体金免疫层析法(LFA)、隐球菌培养和宏基因组二代测序(mNGS)进行检测,比较不同检测方法 对CM的诊断价值.结果 47份CM样本中使用抗真菌药物18份(38.30％),未使用29份(61.70％).2组间mNGS和隐球菌培养的阳性率差异有统计学意义(P<0.05).墨汁染色、LFA、隐球菌培养和mNGS的敏感度分别为51.06％、89.36％、79.31％和74.47％;特异度分别为100％、97.5％、100％和100％.墨汁染色的敏感度和准确度与其他3种检测方法 比较差异均有统计学意义(P<0.05),其他检测方法 间比较差异均无统计学意义(P>0.05),仅墨汁染色与LFA的阴性预测值差异有统计学意义(P<0.05),其他检测方法 之间比较差异均无统计学意义(P>0.05).此外,mNGS还在部分样本中检测到其他病原体.结论 LFA的敏感度最高,建议临床首选;墨汁染色操作简单但敏感度较低,需与其他方法 联合使用;隐球菌培养和mNGS对抗真菌药物敏感,需在使用抗真菌药物前进行检测;mNGS可用于CM的早期诊断,还可以检测或排除合并感染.

Objective:To establish a simple and quick method to explore phagocytosis of B lymphocytes under immunofluorescence microscopy based on cell counting chamber.By monitoring the sample quality before flow cytometry(FCM),the method could improve the stability of phagocytic rate detected by FCM.It could also offer image proof under fluorescence microscopy at the same time.Methods:After Raji B lymphocytes and FITC-stained BCG were incubated together for 24 hrs,B lymphocytes were labeled by PE anti-human CD19 antibody.Then we applied cell counting chamber to observe phagocytosis of B lymphocytes and flow cytometry to test phagocytic rate.Results:B lymphocytes and BCG could be single labeled respectively and double labeled,which was observed under fluorescence microscope based on cell counting chamber.The phagocytic rate tested by flow cytometry was 13.9 ％.Conclusions:Observing phagocytosis of B lymphocytes using cell countirg chamber under fluorescence microscopy might be a simple and quick method.It can be applied in monitoring the sample quality before flow cytometry,it could also offer proof under fluorescence microscopy.

Early diagnosis and treatment are crucial for the outcome of central nervous system (CNS) infections. In this study, we developed a multiplex PCR-Luminex assay for the simultaneous detection of five major pathogens, including Mycobacterium tuberculosis, Cryptococcus neoformans, Streptococcus pneumoniae, and herpes simplex virus types 1 and 2, which frequently cause CNS infections. Through the hybridization reaction between multiplex PCR-amplified targets and oligonucleotide “anti-TAG” sequences, we found that the PCR-Luminex assay could detect as low as 101–102 copies of synthetic pathogen DNAs. Furthermore, 163 cerebrospinal fluid (CSF) specimens from patients with suspected CNS infections were used to evaluate the efficiency of this multiplex PCR-Luminex method. Compared with Ziehl-Neelsen stain, this assay showed a high diagnostic accuracy for tuberculosis meningitis (sensitivity, 90.7% and specificity, 99.1%). For cryptococcal meningitis, the sensitivity and specificity were 92% and 97.1%, respectively, compared with the May Grunwald Giemsa (MGG) stain. For herpes simplex virus types 1 and 2 encephalitis, the sensitivities were 80.8% and 100%, and the specificities were 94.2% and 99%, respectively, compared with Enzyme Linked Immunosorbent Assay (ELISA) assays. Taken together, this multiplex PCR-Luminex assay showed potential efficiency for the simultaneous detection of five pathogens and may be a promising supplement to conventional methods for diagnosing CNS infections.

BACKGROUND:The early detection and identification of pathogens in central nervous system viral infections associated with neurological disease increases the survival rate. However, the limitations of current diagnostic methods contribute to a lack of proper diagnosis in 62% of patients. Therefore, a robust method for detecting multiple viruses in a single reaction with high specificity, throughput, and speed is required.METHODS:A multiplex PCR Mag-Array (MPMA) system was developed that integrates three strategies: chimeric primer design, temperature switch PCR, and MagPlex-TAG techniques. The MPMA was used to amplify 13 target viral sequences simultaneously, with plasmids containing specific viral sequences as standard samples. To evaluate its clinical performance, 177 cerebrospinal fluid (CSF) samples were tested.RESULTS:The MPMA system presented high specificity and efficiency in detecting a control panel of 13 plasmids. Among 177 CSF samples, consistent results were achieved for 19 samples pre-tested using a commercial kit. Viral pathogens were found in 28/138 undiagnosed samples, with herpes simplex viruses (HSV-1 and HSV-2) being predominant. The 20 non-infectious samples revealed negative results. Compared to sequencing methods, sensitivity for detecting HSV-1 and HSV-2 was 100% and 98.78%, respectively, and specificity was 100% and 98.22%, respectively.CONCLUSIONS:A robust MPMA system that can simultaneously and reliably detect 13 meningoencephalitis-associated viruses with high specificity, throughput, and speed has been developed.

Infrasound, a kind of common environmental noise and a major contributor of vibroacoustic disease, can induce the central nervous system (CNS) damage. However, no relevant anti-infrasound drugs have been reported yet. Our recent studies have shown that infrasound resulted in excessive microglial activation rapidly and sequential inflammation, revealing a potential role of microglia in infrasound-induced CNS damage. Epigallocatechin gallate (EGCG), a major bioactive component in green tea, has the capacity of protecting against various neurodegenerative diseases via an anti-inflammatory mechanism. However, it is still unknown to date whether EGCG acts on infrasound-induced microglial activation and neuronal damage. We showed that, after 1-, 2- or 5-day exposure of rats to 16 Hz, 130 dB infrasound (2 h/day), EGCG significantly inhibited infrasound-induced microglial activation in rat hippocampal region, evidenced by reduced expressions of Iba-1 (a marker for microglia) and proinflammatory cytokines (IL-1β, IL-6, IL-18 and TNF-α). Moreover, infrasound-induced neuronal apoptosis in rat hippocampi was significantly suppressed by EGCG. EGCG also inhibited infrasound-induced activation of primary microglia in vitro and decreased the levels of proinflammatory cytokines in the supernatants of microglial culture, which were toxic to cultured neurons. Furthermore, EGCG attenuated infrasound-induced increases in nuclear NF-κB p65 and phosphorylated IκBα, and ameliorated infrasound-induced decrease in IκB in microglia. Therefore, our study provides the first evidence that EGCG acts against infrasound-induced neuronal impairment by inhibiting microglia-mediated inflammation through a potential NF-κB pathway-related mechanism, suggesting that EGCG can be used as a promising drug for the treatment of infrasound-induced CNS damage.

The PCR products containing 81bp rifampicin resistance-determining region(RRDR) of 102 Mycobacterium tuberculosis isolates,including 63 strains from pulmonary tuberculosis patients and 39 strains from tubercular meningitis patients,were analyzed by sequencing. Among the 102 rifampin-resistant strains,mutations were found in the RRDR of 100 strains.The most frequent mutation rates of the rpoB gene of 63 pulmonary tuberculosis associated strains occurred at codons 531(53.97%),526(20.63%),and 516(6.45%).The most common mutation rates of the 39 meningitis tuberculosis associated strains occurred at codons 531(61.54%),526(20.51%),and 533(7.69%). There was no significant difference in the rpoB gene mutation at the 531 and 526 codons of rifampicin-resistant Mycobacterium tuberculosis between tuberculosis meningitis and pulmonary tuberculosis.However,the Ser→Leu mutation at the codon 531 showed to be predominant.

A rapid and simple method for the detection of drug-resistant Mycobacterium tuberculosis is critical for the efficient treatment and control of this pathogen in developing country. Here we developed a single multiplex amplification refractory mutation system (M-ARMS) PCR, in which chimeric-primer and temperature switch PCR (TSP) strategy were included. Using this method, we detected rifampin resistance-associated mutations at codons 511, 516, 526 and 531 in the rifampin resistance-determining region of rpoB gene. The performance of M-ARMS-PCR assay was evaluated with 135 cultured isolates of M. tuberculosis. The sensitivity and specificity were 94.2% and 100%, respectively, compared with direct DNA sequencing, and 86.67% and 89.71%, respectively, compared with culture-based phenotypic drug susceptibility testing. Therefore, this newly-developed M-ARMS-PCR method is useful and efficient with an intended application in provincial Centers for Disease Control and Prevention for rapid detection of rifampin resistance-associated mutations.

Published Ahead of Print 11 January 2012. 10.1128/JCM.05756-11. 2012, 50(4):1166. DOI: J. Clin. Microbiol. Gang Zhao Wen Dai, Ting-Ting Liu, Ying He, Jin-Ge Li, Xiao-Ke Hao and Wang, Xiao-Dan Shi, Lei Ma, Xue-Dong Liu, Yi-Ning Yang, Ping Chen, Ming Shi, Guo-Dong Feng, Jia-Yun Liu, Bing-Ju Cerebrospinal Fluid in Detection of Extracellular M. tuberculosis Improving Mycobacterium tuberculosis and Intracellular De Novo Identifying A Highly Efficient Ziehl-Neelsen Stain: