Predictive value of vitamin D,sex hormones and their receptors on relapse in male patients with clinically isolated syndrome / 重庆医学

Objective To investigate the predictive value of vitamin D,sex hormones and their receptors in the development and regression of male patients with clinical isolated syndrome(CIS).Methods A total of 40 male CIS patients were included in the CIS group and 30 healthy subjects matched in age and long-term res-idence were included in the control group.The levels of vitamin D[1,25(OH)2D2 and 1,25(OH)2D3]inpe-ripheral blood of the included individuals and the first relapsed multiple sclerosis patients(MS group)were detected by ultra-high-performance liquid chromatography-tandem mass spectrometry.The levels of testoster-one(T),progesterone(P),and estradiol(E2)in peripheral blood were detected by electrochemiluminescence.Estrogen receptor(ERα,ERβ),androgen receptor(AR)and vitamin D receptor(VDR)levels in lymphocyte supernatant were detected by enzyme-linked immunosorbent assay(ELISA).Results After a median follow-up of 31.5 months,29 patients(72.5％)with CIS were converted to multiple sclerosis.Compared with pa-tients with non-recurrent CIS patients,patients with multiple sclerosis had a younger age of onset and were prone to positive oligoclonal bands.Compared with HC group,vitamin D levels were reduced in CIS patients,and the difference was statistically significant(P＜0.05).When CIS patients reverted to multiple sclerosis,se-rum levels of E2,ERα,and ERβ were elevated compared with those before relapse,and the levels of testosteroneand AR were decreased,and the difference was statistically significant(P＜0.05).Correlation analysis showed no correlation between peripheral blood levels of vitamin D,sex hormones and their receptors and EDSS in CIS patients.Conclusion Male CIS patients with younger onset age and positive oligoclonal bands are prone to transition to multiple sclerosis.The decrease of vitamin D level may be associated with the onset of male CIS.Vitamin D,sex hormones,and their receptors have no predictive value in the conversion of CIS to multiple sclerosis.

Mechanism of Apolipoprotein E affecting the expression of Matrix Metalloproteinase-9 Astrocytes / 中风与神经疾病杂志

@#Objective　To explore the mechanism of apolipoprotein E （ApoE） affecting the expression of matrix metalloproteinase-9 （MMP-9） in astrocytes. Methods　（1） Astrocytes in wild-type （WT） and ApoE gene knockout （ApoE-/-） C57BL / 6J suckling mice were cultured in vitro；glial fibrillary acidic protein （GFAP） antibody were employed to identified the astrocytes. （2） Astrocytes from wild type WT and ApoE-/- C57BL/6J suckling mice were divided into blank control group（Control group），negative control group（NC group） and p65 gene knock down group（KD group）. No lentivirus was added to the Control group. The negative control lentivirus was added to the NC group，and the positive lentivirus that silended the p65 gene was added to the KD group. The nuclear factor-κB （NF-κB） p65 gene of astrocytes was silenced with siRNA lentivirus to inhibit NF-κB signal transduction. The expression of p65 protein was detected by Western blot，and MMP-9 and tumor necrosis factor-α（TNF-α） were detected by ELISA. （3）Add TNF-α to stimulate the astrocytes of each group for 24 hours，and the concentration of MMP-9 in each group of astrocytes before and after stimulation was measured by ELISA. Results　（1） The expression of NF-κB p65 protein and the concentrations of MMP-9 and TNF-α in the two astrocytes in the KD group were significantly lower than those in the Control group and the NC group （P<0.05），but those between the Control group and the NC group were no significant different（P>0.05）；（2） The concentration of MMP-9 and TNF-α of ApoE-/- astrocytes were higher than WT astrocytes in the Control group and NC groups （P<0.05），while those was no significant difference between ApoE-/- astrocytes and WT astrocytes in KD group （P>0.05）；（3） The concentrations of MMP-9 of the two types of astrocytes in Control group and NC group was increased after the stimulation of TNF-α（P<0.05），while the concentration of MMP-9 in the KD group had no significant change（P>0.05）. Conclusion　ApoE may affect the expression of MMP-9 in astrocytes through the TNF-α/NF-κB signaling pathway，and then affects the integrity of the blood-brain barrier.

Effects of apolipoprotein E deficiency on neuromyelitis model in vitro / 中华神经科杂志

Objective To investigate the effects of apolipoprotein E (ApoE) deficiency on neuromyelitis optica (NMO) model of spinal cord sections induced by NMO-IgG and complement in vitro.Methods NMO-IgG was extracted from the patients with NMO,and complementary serum from healthy people.The spinal cord sections of seven days old C57BL / 6J mice with wild type (WT) or ApoE knockout (ApoE-/-) were cultured for seven days.The spinal cords of the two genotypes were randomly divided into experimental groups (NMO-ApoE-/-group,NMO-WT group) and control groups (C-AopE-/-group,C-WT group),respectively.The experimental groups were treated with NMO-IgG and complementary serum,and the control groups only with complementary serum.Then all the sections were continued incubating for 24 h before harvested.Immunofluorescence staining and modified thick tissue film immunofluorescence were used to detect the expression of aquaporin 4 (AQP4),glial fibrillary acidic protein (GFAP),ionic calcium fibronectin (IBA1),myelin basic protein (MBP) and human neurofilament protein L (NFL) respectively.The lesion score was calculated according to the areas percentage of AQP4 and GFAP deficiency in spinal cord sections.Results Compared with the respective control groups,the expressions of AQP4,GFAP,MBP and NFL were deficient in the experimental groups (The percentages of missing area in the NMO-ApoE-/-group were 83.88％ ± 5.01％,82.44％ ± 6.11％,45.02％ ± 5.11％ and 54.65％ ± 7.66％ respectively,while the percentages of missing area in the C-ApoE-/-group were 10.44％ ± 4.07％,5.73％ ±0.82％,9.12％ ±1.41％ and 5.72％ ±0.81％,t=34.143,37.269,20.300,19.051,allP ＜0.05;The percentages of missing area in the NMO-WT group were 77.74％ ± 6.75％,75.62％ ± 5.76％,37.60％ ± 4.88％ and 46.29％ ± 4.98％,while the percentages of missing area in the C-WT group were 9.31％ ± 2.97％,5.80％ ± 0.82％,9.10％ ± 1.63％,5.80％ ± 0.81％ respectively,t =27.828,35.934,16.613,24.057,all P ＜ 0.05).While IBA1 was up-regulated and the damage scores were higher in both the NMO-ApoE-/-group and the NMO-WT group.The percentages of missing area in the NMO-ApoE-/-group and the NMO-WT group showed statistically significant difference (t =2.194,2.436,3.149,2.746,all P ＜ 0.05).The expression level of IBA1 in the NMO-WT group was higher than that in the C-WT group (19.88 ± 1.11 vs 11.18 ±0.65,t =25.270,P ＜0.05),while the expression level of IBA1 in the NMO-ApoE-/-group was higher than that in the NMO-WT group (25.81 ± 1.61 vs 19.88 ± 1.11,t =9.101,P ＜0.05).The degree of deficiency or up-regulation of above-mentioned proteins was more obvious in the NMO-ApoE-/-group than that in the NMO-WT group.Conclusions NMO-IgG extracted from NMO patients can induce NMO-like damage in isolated tissue at the presence of complement.ApoE deficiency promotes the further activation of microglia,thereby aggravates the injury of astrocyte in the model of NMO.

Management of sputum disposal in tuberculosis ward based on PDCA circulation method / 中国实用护理杂志

Objective To improvethe control of infection source by improving the continuous quality of sputum disposal management. Methods PDCA method based on the management of tuberculosis sputum disposal of the hospital quality improvement. The cross-sectional survey of tuberculosis ward in 32 cases of hospital patients, as control group.PDCA first round of "sputum cup distribution and use of work flow" for quality improvement,the reason leading the work flow of workers to nurse led work flow,the improved cross-sectional survey in 36 cases of hospital patients as experimental group 1.Aiming at the problem of low utilization rate of special sputum cup in the improvement of the first round of PDCA,the improvement of the sputum cup was carried out,On this basis, the "special sputum disposal Cup" was designed to obtain the national patent authorization. After the second round of improvement, the cross-sectional investigation in 35 patients in the hospital,as the experimental group 2.Results After two rounds of PDCA improvement,experimental group 2 groups compared with control,bedside sputum cup configuration was not in place rate decreased from 53.1% (17/32)to 0,the difference was statistically significant(Χ2=24.916,P<0.05);configuration after the sputum cup unused rate from 53.1%(17/32)down to 8.6%(3/35),the difference was statistically significant(Χ2=15.846,P<0.05).Conclusion PDCA method can improve the quality of management standard sputum disposal. Ward sputum cup ration is standardized sputum disinfection ward-based nurse intervention and leading sputum cup release, ward can improve the sputum cup ration, improved sputum containers (sputum cup) can improve patient compliance standard of spitting.

Effect of apolipoprotein E on secretions of matrix metalloproteinase-9, matrix metalloproteinase tissue inhibitor-1 and inflammatory factors in macrophages at early stage of experimental autoimmune encephalomyelitis / 中华神经医学杂志

Objective To explore the effect of apolipoprotein E (ApoE) on secretions of matrix metalloproteinase-9 (MMP-9),matrix metalloproteinase tissue inhibitor-1 (TIMP-1) and inflammatory factors in spleen macrophages at early stage of experimental autoimmune encephalomyelitis (EAE).Methods In accordance with random number table,20 ApoE knockout (ApoE-/-) C57BL/6J mice were divided into ApoE-/-EAE group and ApoE-/-control group (n=10);and 20 wild type (WT) C57BL/6J mice were divided into WT EAE group and WT control group (n=10).Myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) was used as antigen to induce EAE models in the ApoE-/-EAE group and WT EAE group,and MOG35-55 was replaced by normal saline in ApoE-/-control group and WT control group.The spleen macrophages were isolated from mice after 7th d of modeling and cultured in vitro;the rnacrophages of the ApoE-/-EAE group and WT EAE group were stimulated by MOG35-55,while the macrophages of the ApoE-/-control group and WT control group were treated with equivalent normal saline.Forty-eight hafter that,the contents of MMP-9,TIMP-1,interferon gamma-γ(IFN-γ),tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the culture medium were determined by ELISA.Results The contents of MMP-9,TFN-γ TNF-α and IL-6 in the WT EAE group were significantly higher than those in the WT control group (P＜0.05);the contents of MMP-9,IFN-γ 、TNF-α and IL-6 in the ApoE-/-EAE group were significantly higher than those in the ApoE-/-control group (P＜0.05);the content ofMMP-9,IFN-γ,TNF-cα and IL-6 in ApoE-/-EAE group were significantly higher than those in the WT EAE group (P＜0.05).The TIMP-1 secretion ofmacrophages in WT EAE group was significantly higher than that in WT control group (P＜0.05).Conclusions At early stage of EAE,macrophages secrete high levels of MMP-9 and inflammatory factors,and the deficiency of ApoE further contributes to the secretions of MMP-9 and inflammatory factors by macrophages.ApoE may play a role in protecting the integrity of blood-brain barrier and alleviating the inflammatory response of the central nervous system through the effects on macrophages.

Effect of apolipoprotein E on matrix metalloproteinases-9 expression and its mechanism in astrocytes / 中华神经医学杂志

Objective To explore the effect of apolipoprotein E (ApoE) on matrix metalloproteinases-9 (MMP-9) expression and its mechanism in astrocytes.Methods (1) Astrocytes in ApoE gene knock out (ApoE-/-) and wild type (WT) C57BL/6J suckling mice were cultured in vitro;glial fibrillary acidic protein antibodies were employed to identify the astrocytes;MMP-9.antibodies were employed to detect the MMP-9 expression in the astrocytes.(2) Astrocytes from the ApoE-/-and wild type WT C57BL/6J suckling mice were divided into activation group,antibody groups and control group randomly;cells in the control group did not give any treatment;cells in the activation group were given whooping cough toxin and thermal inactivation-mycobacterium tuberculosis H37Ra;cells in the antibody groups were given anti-interleukin-6 (IL-6),interferon gamma (IFN--γ),tumor necrosis factor-α (TNF-α) and IL-12,respectively,to inhibit their inflammatory factors.ELISA was performed to detect the concentrations of MMP-9 and tissue inhibitor of metalloproteinase-1 (TIMP-1),and the inflammatory factors concentrations ofTNF-,IL-12,IL-6,and IFN-γ.Results These two kinds of rat astrocytes could both qualitatively express MMP-9;as compared with the control group,the activation group had significantly higher MMP-9 concentration (P＜0.05);the activation group had significantly higher MMP-9 concentration as compared with the antibody groups (P＜0.05);activation group from ApoE-/-C57BL/6J suckling mice had significantly higher MMP-9 concentration than that from wild type C57BL/6J suckling mice (P＜0.05).The concentration of TIMP-1 was not significantly different among various groups (P＞0.05).The concentrations of inflammatory factors in the activation groups from two kinds of mice were significantly higher than those in the control group (P＜0.05);the concentrations of IL-6,IFN-γ TNF-α and IL-12 in each antibody group were significantly lower than those in the activated group (P＜0.05);the concentrations of inflammatory factors in ApoE-/-rats were significantly higher than those in WT rats (P＜0.05).Conclusion ApoE can regulate the MMP-9 expression by regulating the secretion of proinflammatory cytokines,which can affect the integrity of blood brain barrier.

"Flipped classroom" teaching model into the curriculum of:exploration and practice / 中国针灸

The "flipped classroom" teaching model practiced in the teaching ofcurriculum was introduced. Firstly, the roles and responsibilities of teachers were clarified, indicating teachers provided examples and lectures, and a comprehensive assessment system was established. Secondly, the "flipped classroom" teaching model was split into online learning, classroom learning and offline learning. Online learning aimed at forming a study report by a wide search of relevant information, which was submitted to teachers for review and assessment. Classroom learning was designed to communicate study ideas among students and teachers. Offline learning was intended to revise and improve the study report and refined learning methods. Lastly, the teaching practice effects of "flip classroom" were evaluated by comprehensive rating and questionnaire assessment, which assessed the overall performance of students and overall levels of paper; the learning ability was enhanced, and the interest and motivation of learning were also improved. Therefore, "flipped classroom" teaching mode was suitable for the curriculum of, and could be recommended into the teaching practice of related curriculum of acupuncture and tuina.

Therapeutic effect observation on Nie-pinching the spine manipulation for infantile diarrhea due to spleen deficiency / 针灸推拿医学（英文版）

Objective: To observe the influence of Nie-pinching the spine manipulation on the excretory rate of urine D-xylose in the infants with diarrhea due to spleen deficiency, and to assess the clinical effects. Methods:Sixty infants in conformity with the diagnostic criteria of diarrhea due to spleen deficiency were randomly divided into a treatment group and a control group by the random digital table, 30 cases in each group. The infants in the treatment group were treated by Nie-pinching the spine manipulation and traditional infantile tuina, in addition to the routine basic treatment. The infants in the control group were treated by the same traditional infantile tuina in addition to the routine basic treatment. The infants in the two groups were treated once every day, 4 weeks as a course. Totally, the treatment was given for a course. The symptom integrals of spleen deficiency were used to assess the improvement in the symptoms. The colorimetry was used to determine the excretory rate of urine D-xylose. Results:In the comparison of the same group before and after the treatment, the differences in the global score of spleen deficiency symptoms and the excretory rate of urine D-xylose in the two groups were statistically significant (allP<0.01). After the treatment, the differences in the global score of spleen deficiency symptoms and the excretory rate of urine D-xylose between the two groups were all statistically significant (bothP<0.01). Conclusion:Chiropractics can reduce the integrals of spleen deficiency symptoms and elevate the excretory rate of urine D-xylose in the infants with diarrhea due to spleen deficiency, so as to enhance the therapeutic effects by alleviating the symptoms of spleen deficiency and the absorptive function of the small intestine.

Synthesis and protein tyrosine kinases inhibitory activity of aroyl derivatives / 国际药学研究杂志

Objective Using Ex-Rad as a lead compound to design and synthesize aroyl derivatives with protein tyrosine ki?nases(PTK)inhibitiory activity. Methods 1-[(4-Fluorophenyl)amioncarbonyl]cyclopropanecarboxylic acid,and 2-oxo-1-phenyl-imidazolidine were used as raw materials to synthesize intermediates 3a-3d,respectively. The target compounds T1-T7 were synthe?sized by chloroformylation reaction with 3a-3d. Enzyme-linked immunosorbent assay(ELISA)was used and inhibitory rate was calcu?lated to screen out the compounds with PTK inhibitory activity. Results Seven new compounds containing aroyl groups were synthe?sized and their structures were confirmed by 1H NMR. The evaluation of the seven compounds demonstrated that PTK inhibitory activi?ty of T2 and T6 were stronger than that of the lead compound. Conclusion The synthetic method is simple,and the materials are cheap and readily available. T2 and T6 show strong PTK inhibitory activity by ELISA.

The relationship between the neuromyelitis optica astrocyte damage and the patients with functional defect / 实用医学杂志

Objective To explore the relationship between the damage of neuromyelitis optica (NMO) astrocytes (AS) and the onset of NMO, and investigate the relevance of AS damage with the severity of the patients with functional defect. Methods The levels of aquaprin4-antibody (AQP4 -Ab), glial fibrillary acidic protein (GFAP), apolipoprotein(ApoE),interleukins-6(IL-6), interleukins-10(IL-10), tumor necrosis factor-α(TNF-α) in cerebrospinal fluid (CSF ) and serum of 30 acute NMO patients were tested by means of ELISA. The results were later compared with control group. And analysis of the relevance of the various index of the levels in CSF with the CSF AQP4-Ab level, the acute phase expanded disability status scale(EDSS) score of the NMO group were made. Results (1)The NMO group in CSF and serum AQP4-Ab, GFAP, IL-6 levels were higher than the control group (P < 0.05), and ApoE, IL-10 levels were lower than the control group (P < 0.05). (2)The CSF GFAP, ApoE, IL-6 in NMO group is higher than the serum (P < 0.05), and CSF AQP4-Ab, IL-10 levels were lower than the serum ( P < 0 . 05 ) . ( 3 ) The CSF GFAP , IL-6 levels and the CSF AQP4-Ab level were positively correlated (r=0.749, r=0.526, P<0.05), and the CSF ApoE, IL-10 levels were negatively correlated with CSF AQP4-Ab level(r = -0.571, r = -0.676, P < 0.05). (4)The CSF AQP4-Ab, GFAP, IL-6 levels and the acute phase EDSS score were positively correlated (P < 0.05), the CSF ApoE, IL-10 levels were negatively correlated with the acute phase EDSS score (P < 0.05). Conclusion The AS damage exists in the NMO and the damage severity may correlate with patient function defect. AQP4-Ab, GFAP, IL-6 may play important roles in the onset of NMO and the disease aggravating. The decrease of the ApoE and IL-10 may exacerbate NMO damage.

Effect of apolipoprotein E deficiency on astrocyte destruction / 实用医学杂志

Objective To explore the effect of apolipoprotein E deficiency on astrocyte destruction in the ex vivo spinal cord slice model. Methods Vibratome-cut transverse spinal cord slices from C57BL/6 postnatal day 7 ApoE -/-mice and wild-type mice were cultured on transwell porous supports. After 7 days in the culture , spinal cord slices were exposed to LPS for 5 days. The expression of AQP4, GFAP and Iba1 was detected by immunofluorescence. The concentration of TNF-α and NO were detected by ELISA and method of nitrate reductase, respectively. Results Marked loss of AQP4 and GFAP staining were shown in LPS-affected groups, not in the control group, and the lesion score was higher in ApoE-/-group than WT-group (P<0.05). In addition, the expression of Iba1 and concentration of TNF-α, NO in the LPS-affected groups were more than that of the control group (P < 0.05), and they were higher in the ApoE-/-LPS group than the WT-LPS group (P < 0.05). Conclusions ApoE deficiency exacerbates astrocyte injury, likely through promoting the release of pro-inflammatory mediators from microglia.

Neuronal damage and neurite change in cell model of intractable epilepsy / 中华神经科杂志

Objective To establish the cell model of intractable epilepsy and to observe its neuronal damage and morphologic change of neurites.Methods The model was established by exposing hippocampal neurons to Mg2+ -free media for 3 hours on days 10 of culture.Expression of lactic acid dehydrogenase (LDH) in supernatant was measured as an index of neuronal damage.The morphologic change of neurons and neurites was observed by optical microscope and scanning electron microscope (SEM).Results Compared to the control group, level of LDH (U/L) was significantly increased in the model group at different time points (3 hours: 4.26 ± 1.28, 6 hours: 6.56 ±2.34 and 24 hours: 16.67 ±3.57, P <0.05).With time prolonging, release of LDH in the model group was gradually increased (F = 39.316,P <0.05).Under optical microscope, neurons of model group migrated closely to each other and neurite connections appeared to be gradually "reticulated" after Mg2+ -free media treatment for 24 hours; and the "reticulated" neurites connections become more obvious after 72 hours.Under SEM, neuronal membrane was rough and had several small depressions, neurites were interlaced in cluster.Conclusions Neuronal damage and morphologic change of neurites are verified in the cell model of intractable epilepsy.

Effects of glucocorticoid on the expression of CD80 and CD4~+CD25~+T cells in peripheral blood lymphocyte of patients with multiple sclerosis / 临床神经病学杂志

Objective To investigate the effects of glucocorticoid(GC) on the expression of CD80 and CD4+CD25+T cells in peripheral blood lymphocyte of patients with multiple sclerosis (MS). Methods With flow cytometer,the positive rates of CD80 and CD4+CD25+T cells in peripheral blood lymphocyte were detected in 21 acute MS patients pre and post treatment separately,and compared with the nomal control group.The secols of Expanded Disablility Status Scale(EDSS) were compared pre and post treatment in MS patients.Results The positive rate of CD80[(5.031?1.782)%]in the MS patients pre treatment was obviously less than that in control group's [(6.436?2.035)%](P