Associations between dysbiosis gut microbiota and changes of neurotransmitters and short-chain fatty acids in valproic acid model rats.

Introduction: The microbiota-gut-brain axis plays an important role in the pathophysiology of autism spectrum disorder, but its specific mechanisms remain unclear. This study aimed to explore the associations of changes in neurotransmitters and short-chain fatty acids with alterations in gut microbiota in valproic acid model rats. Methods: The autism model rats were established by prenatal exposure to valproic acid (VPA). The Morris water maze test, open field test, and three-chamber test were conducted to assess the behaviors of rats. 16S rRNA gene sequences extracted from fecal samples were used to assess the gut microbial composition. Gas and liquid chromatography-mass spectroscopy was used to identify short-chain fatty acids in fecal samples and neurotransmitters in the prefrontal cortex (PFC). Results: The results showed that 28 bacterial taxa between valproic acid model rats and control rats were identified, and the most differential bacterial taxa in valproic acid model rats and control rats belonged to metagenomic species and Lactobacillus intestinalis. Acetic acid, butyric acid, valeric acid, isobutyric acid, and isovaleric acid were significantly decreased in the valproic acid model rats compared to those in control rats. Five neurotransmitters (threonine, kynurenine, tryptophan, 5-hydroxyindoleacetic acid, denoted as 5-HIAA, and betaine aldehyde chloride, denoted as BAC) were significantly decreased, whereas betaine was increased in the prefrontal cortex of valproic acid model rats compared to control rats. A variety of neurotransmitters (≥4) were correlated with Pseudomonas, Collisella, and Streptococcus at the genus level, and they were also related to the decrease of short-chain fatty acids. Discussion: According to this study, we can preliminarily infer that gut microbiota or their metabolic productions (such as SCFAs) may influence central neurotransmitter metabolism through related pathways of the gut-brain axis. These results provide microbial and short-chain fatty acid (SCFA) frameworks for understanding the role of the microbiota-gut-brain axis in autism spectrum disorder and shed new light on autism spectrum disorder treatment.

Mesenchymal stem cells alleviate AQP-4-dependent glymphatic dysfunction and improve brain distribution of antisense oligonucleotides in BACHD mice.

Huntington's disease (HD) is a neurodegenerative disorder caused by a mutation in the huntingtin (HTT) gene that results in the production of neurotoxic mutant HTT (mHTT) protein. Suppressing HTT production with antisense oligonucleotides (ASOs) is a promising treatment strategy for HD; however, the difficulty of delivering ASOs to deep brain structures is a major barrier for its clinical application. The glymphatic system of astrocytes involving aquaporin 4 (AQP-4) controls the entry of macromolecules from the cerebrospinal fluid into the brain. Mesenchymal stem cells (MSCs) target astrocytes to inhibit neuroinflammation. Here we examined the glymphatic distribution of ASO in the brain and the therapeutic potential of combining intravenously injection of mesenchymal stem cells (IV-MSC) and ASOs for the treatment of HD. Our results show that Cy3-labeled ASOs entered the brain parenchyma via the perivascular space following cisternal injection, but the brain distribution was significantly lower in AQP-4-/- as compared with wild-type mice. Downregulation of the AQP-4 M23 isoform was accompanied by decreased brain levels of ASOs in BACHD mice as well as an increase in astrogliosis and phosphorylation of nuclear factor κB (NF-κB) p65. IV-MSC treatment restored AQP-4 M23 expression, attenuated astrogliosis, and decreased NF-κB p65 phosphorylation; it also increased the brain distribution of ASOs and enhanced the suppression of mHTT in BACHD mice. These results suggest that modulating glymphatic activity using IV-MSC is a novel strategy for improving the potency of ASO in the treatment of HD.

Rifampicin attenuates experimental autoimmune encephalomyelitis by inhibiting pathogenic Th17 cells responses.

Rifampicin, a broad-spectrum antibiotic, has neuroprotective, immunosuppressive, and anti-inflammatory properties. However, the effect of rifampicin on autoimmune disorders of the nervous system is not clear. In this study, we investigated whether rifampicin was beneficial to myelin oligodendrocyte glycoprotein peptide (MOG33-35 )-induced female C57BL/6 experimental autoimmune encephalomyelitis (EAE) mice, the well-established animal model of multiple sclerosis. Rifampicin treatment (daily from the first day after EAE immunization) remarkably attenuated clinical signs and loss of body weight, which are associated with suppression of inflammatory infiltration and demyelination in spinal cords of EAE mice. Furthermore, rifampicin dramatically reduced the disruption of blood-brain barrier integrity, down-regulated serum concentration of IL-6 and IL-17A, inhibited pathological Th17 cell differentiation, and modulated the expression of p-STAT3 and p-p65. These results suggest that rifampicin is effective for attenuating the clinical severity of EAE mice, which may be related to its inhibitive ability in differentiation of Th17 cell and secretion of its key effector molecule IL-17A via regulation of excessive activation of the key signaling molecules of JAK/STAT pathway. Our findings may be helpful for developing therapeutic and preventive strategies for multiple sclerosis.

[Quantitative detection of serum procollagen III N-terminal peptide chemiluminescence enzyme immunoassay].

OBJECTIVE: To establish chemiluminescence enzyme immunoassay (CLEIA) for quantitative detection of procollagen III N-terminal peptide (P III NP) in serum. METHODS: A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of P III NP as the catalytic enzyme and the luminol as the luminescence reagent. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on. The CLEIA was compared with imported ELISA kits, by detecting clinical serum. RESULTS: The linear range was 0.8-85 ng/ml. The detection limit was 0.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 96.2%, 91.2% and 101.1%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.99 and RSD lower than 6%. The detected results of clinical sera with CLEIA closely corresponded to those with imported ELISA. CONCLUSION: Established CLEIA for quantity determination of serum P III NP has high accuracy, sensitivity and repeatability.

A 65-year-old man with infratentorial diffuse encephalopathy and hydrocephalus.

Tuberculous encephalopathy (TBE) is an important diagnosis in countries with a high prevalence of tuberculosis. TBE is a life-threatening condition but rarely reported in the modern literature. We reported a case of a man with extensive parenchymal lesions involving the brainstem and right cerebellar hemisphere that resolved after treatment. The clinical, laboratory and pathological features of this case are highlighted and the pathogenesis is discussed.

[Analysis for post-transplant changes of peripheral lymphocyte subsets in liver transplant receptors with HBV infection].

AIM: To investigate the characters and changes of peripheral lymphocyte subsets and cytokines in liver transplant receptors with HBV infection in short phases after liver transplantation, and to provide evidences for monitoring post-transplant immune condition of liver transplant receptors. METHODS: Peripheral lymphocyte subsets and cytokine levels in pre- and post- transplant 12 h, 3 d, 10 d, 30 d, 60 d of 20 cases of patients with HBV-associated severe hepatic diseases were investigated and analyzed, and were compared respectively with those of 22 cases of healthy adults as control (HC) with flow cytometry (FCM) and ELISA. RESULTS: The patients' accounts of peripheral lymphocyte subsets before liver transplantation were lower than those of HC significantly, but the accounts decreased significantly after transplantation 12 h. Three days later, the accounts of lymphocyte subsets increased significantly. The percentages of CD3, CD4, CD8 and NK cells got to stable stage from post-transplantation 10 d, and the absolute accounts of post-transplantation 60 d were higher than those of pre-transplantation, but were still lower than those of HC; The IFN-γ and IL-10 levels of post-transplantation 12 h increased several times and decreased after 3 days. The IL-10 levels in post-transplantation 60 d were still higher than those of HC. CONCLUSION: The absolute accounts of peripheral lymphocyte subsets increased to stable levels from post-transplantation 10 d, but were still lower than those of HC; Post-transplant immune condition was to Th2 polarization.

[Clinical and laboratory characteristics of a new influenza A (HIN1) epidemic].

OBJECTIVE: To analysis the clinical and laboratory characteristics of Patients infected with new influenza A (HIN1) virus. METHODS: All cases with new influenza A (H1N1) confirmed on polymerase chain reaction assay on throat swabs. There were included in a prospective evaluation of clinical characteristics, laboratory results, treatment and overcome of new influenza A (H1N1). RESULTS: There were 35 patients in the epidemic. Clinical illness developed within a mean of 1.7 days. Fever occurred in 97.1%, sore throat 65.7% cough 51.4%, headache 28.6%, and myalgia 31.4%. All patients were treated with oseltamivir lasted 5 days. The mean duration of viral shedding was 4.5 days. All were cured and left hospital after day 7. CONCLUSION: It was infected by new influenza A (H1N1) typically in this epidemic.

Anti-tuberculosis treatment for Devic's neuromyelitis optica.

There are no specific treatments for patients with acute, severe neurological deficits caused by neuromyelitis optica (NMO) who fail to recover after treatment with high-dose corticosteroids. We evaluated the clinical response of anti-tuberculosis treatment (ATT) in patients suffering from steroid-refractory NMO, and investigated the correlation between NMO and tuberculous infection of the central nervous system (CNS). We conducted this prospective, controlled study in southern China. Twelve patients with steroid-refractory NMO were monitored during ATT and compared with a control group of 13 patients with the same type of NMO who received current standard therapies. A molecular diagnostic test was carried out and Extended Disability Status Scale (EDSS) score analysis, visual acuity, the number of relapses and MRI changes were evaluated at study entry and after 1 and 2years of therapy. ATT may lead to the recovery of important neurological functions and all our patients responded positively to therapy. EDSS score and visual acuity improved and abnormalities in the spinal cord, observed by MRI, markedly decreased over time. ATT also significantly reduced the rate of relapse. By comparison, in the control group, a significant clinical deterioration was observed, and patients did not show favourable EDSS scores and MRI changes. This study suggests that CNS infection with Mycobacterium tuberculosis is an important cause of NMO.

[Changes and analysis of peripheral white blood cells and lymphocyte subsets for patients with pandemic influenza A virus (H1N1) infection].

OBJECTIVE: To investigate the characters and changes of peripheral white blood cells and lymphocyte subsets of patients with pandemic influenza A virus (H1N1) infection and to provide evidences for diagnosis, treatment and prognosis of influenza A (H1N1) infection. METHODS: Peripheral white blood cell parameters and the percentages of lymphocyte subsets in acute and recovery phases of 59 cases of influenza A virus (H1N1) infectious patients (42 mild cases and 17 severe cases) were investigated and analyzed, and compared respectively with those of 43 cases of healthy adults as control (HC) and 24 cases of general influenza A virus (no-H1N1) infectious using whole blood cell analysis and flow cytometry. RESULTS: Peripheral white blood cell counts of mild cases decreased greatly but those of severe cases did not decrease significantly; the neutrophils of severe cases increased significantly in acute phase; similar to general influenza A virus (no-H1N1) infectious, the peripheral lymphocytes, CD3, CD4, CD8 and B cells of all patients with influenza A virus (H1N1) infection decreased greatly in acute phase and quickly recovered in recovery phase; NK and NKT cells absolute counts of severe cases decreased significantly in acute phase, and the decreasing extent of which were more than 20%. CONCLUSION: There were similar characteristics of change in peripheral white blood cells and lymphocyte subsets between patients with pandemic influenza A virus (H1N1) infection and general pandemic A virus (No-H1N1); the great decrease of NK and NKT cells absolute counts may suggest the severe tendency of diseases.

[Study of copper metabolism and liver damage in TX Mice-an animal model for liver disease].

OBJECTIVE: To provide right time points in selection of right aged animals and the normal physiological data of TX mice. METHODS: 7-12 months old TX and DL mice were studied, each group contained 3 female and 3 male mice of TX or DL mice. The concentration of copper in the serum, dry tissues (liver, brain and kidney), together with copper biochemistry indexes were measured. The liver histopathology was observed under light microscopy and electron microscope. RESULTS: Transaminase increased significantly only in 10 and 11-month- old (AST(TX10) = 218.3 U/L, AST(TX11) = 197.5 U/L, AST(DL10) = 171.5 U/L, AST(DL11) = 165.0 U/L, P(10) less than 0.001, P(11) = 0.022), but the copper concentration of liver, brain and kidney was significantly increased during 7-12 month old (the average concentration of copper, Liver(TX) = (750.0 +/- 85.5) mg/kg, Brain(TX) = (39.7 +/- 2.2)mg/kg, Kidney(TX) = (29.8 +/- 5.0) mg/kg, Liver(DL) = (11.6 +/- 1.5) mg/kg, Brain(DL) = (16.8 +/- 0.9) mg/kg, Kidney(DL) = (14.2 +/- 1.0) mg/kg, t = 21.16, 23.60, 7.47, for all these organs P less than 0.05). CONCLUSION: TX mice is a suitable model of liver disease with natural recovery, so selecting animal model of suitable time point is very important.

[Clinical manifestations and molecular genetics of spinal bulbar muscular atrophy: report of 5 cases].

OBJECTIVE: To study the clinical and molecular genetic characteristics of spinal bulbar muscular atrophy (SBMA). METHODS: The clinical data, including case history, physical examination, biochemical analyses of blood, EMG, and muscle biopsy, of 5 Chinese patients with SBMA, all males, aged 29 - 58, with the onset age of 36 (17 - 49), were collected the information of in 5 cases. Four patients underwent PCR to examine the number of copies of CAG repeat region in androgen receptor (AR) gene. RESULTS: The clinical characteristics of the 5 patients included atrophy of lingualis, dysarthria, weakness and waste of the limbs, especially in the hands, and elevated creatine kinase (CK), fasting glucose, testosterone, and progesterone in the blood. EMG showed denervation motor potentials in all cases. The muscle biopsy in one case showed neurogenic atrophy. The number of (CAG) n repeat in AR gene was 50 - 62 in the, remarkably from that of 13 normal controls (19 - 20) without overlapping. CONCLUSION: SBMA affects the middle age males, shows a slowly progressing muscular atrophy in spinal and bulbar muscles. The different number of (CAG) n repeat of AR gene between the SBMA patients and the normal controls may be an important identification to differentiate SBMA from other motor neuron diseases.

[Establishment and evaluation of the diagnostic kit for anti-HIV1/2 antibody and P24 antigen].

OBJECTIVE: To establish and evaluate an Enzyme Immunoassay diagnostic kit combined with anti-HIV1/2 antibody and P24 antigen for shortening the examination window period of HIV infection in HIV laboratory diagnosis. METHODS: The enzyme-linked reaction plates was coated by anti-HIV P24 monoclonal antibody and HIV 1/2 antigen. Labeling HIV1/2 antigen and anti-HIV P24 polyclonal antibody with horseradish peroxidase, setup an integrated ELISA kit for detecting anti-HIV-1/2 antibody and HIV P24 antigen, and evaluate the specificity and sensitivity of this kit. RESULTS: The sensitivity of testing P24 antigen was up to 0.2 ng/ml. 78 serum samples of patients with AIDS, 85 serum samples of healthy people were compared with Abbott EIA kit, the coincidence was 100%. 12 051 sera from normal persons and patients were examined, the sensitivity of 100 %and specificity of 99.62 %, respectively. CONCLUSION: The anti-HIV1/2 antibody and HIV P24 antigen can be measured at the same time using this EIA kit, while the examination window period of HIV infection is shortened. Thus, the method is suitable for laboratory diagnosis and epidemiological investigation.

[Study on simultaneous analysis of indoor air multi-component VOCs with FTIR].

Recently, the indoor air pollution, especially the volatile organic compounds (VOCs) is attracting more and more attention. The determination of indoor air organic gases becomes the key issue. In the present paper, a simultaneous quantitative measuring method of indoor air multi-component VOCs was established based on FTIR combined with chemometrics. The 3 200-2 600 cm(-1) and 1 100-600 cm(-1) bands of IR spectrum were used to establish validation model, and excellent coefficients were obtained (r(2) =0. 970, 0. 955 and 0. 946 for benzene, toluene and dimethylbenzene, respectively). The root mean standard error of calibration for benzene, toluene and dimethylbenzene is 0. 074 2, 0. 081 9 and 0. 087 7, respectively. The root mean standard error of prediction is 0. 132, 0. 134 and 0. 033 3, respectively. The error of unknown sample prediction is acceptable. The IR method is effective for simultaneous analysis of indoor air multi-component VOCs. The model established by partial least squares (PLS) is better than that by principal components analysis (PCR).

[Molecular genetic diagnosis and clinical analysis of spinocerebellar ataxia type 7].

OBJECTIVE: To study the molecular genetic diagnosis and clinical characteristics of spinocerebellar ataxia type 7 (SCA7). METHODS: This study included 43 patients with autosomal dominant SCA from 36 families, 38 sporadic SCA patients, 60 healthy individuals from the SCA families and 44 normal controls without family SCA history. The SCA7 (CAG)n mutations were detected by PCR, denaturing polyacrylamide gel electrophoresis and silver staining technique. The abnormal allele fragments were sequenced by ABI373 DNA sequencing machine. RESULTS: Normal alleles of SCA7 were found to have 9 to 19 CAG repeats. Two familial SCA and one sporadic patients were identified by detecting the presence of abnormal CAG-repeat expansion in the SCA7 alleles, which was confirmed by DNA sequencing. The repeats of CAG were 65, 65, and 63 respectively. CONCLUSIONS: Abnormal CAG expansion is the pathogenic cause of SCA7. Molecular genetic analysis is effective for the diagnosis of SCA, prediction of presymptomatic patients and genetic counseling.