Objective:To construct a mouse asthma model induced by mugwort pollen and to explore endotyping,providing methods for subsequent precision treatment.Methods:BALB/c mice were intraperitoneally injected with mugwort pollen extract(MPE)to sensitize,following MPE intranasal challenge to construct MPE allergic asthma murine model.Mice were randomly divided into PBS sensitization and PBS challenge(P-P),MPE sensitization and PBS challenge(M-P),MPE sensitization and MPE challenge model(M-M)groups.24 h after final challenge,mice were performed to examine airway responsiveness;bronchoalveolar lavage fluid(BALF)was harvested for cell counting and statistical classification of inflammatory cells through flow cytometry analysis.Pulmonary slides were collected for pathological examination,including HE,PAS,Masson and α-SMA immunohistochemical staining.ELISA was used to detect levels of IFN-γ,IL-4,IL-5,IL-13,IL-17A in lung tissue and serum,as well as serum total IgE and MPE-specific IgE,IgG1,IgG2a levels.Results:Pathological examination showed higher airway reactivity,more inflammatory cells infiltration around airway,obvious goblet metaplasia,thickening of airway smooth muscles and dramatical fibrin deposition around airway in model group.Total cell numbers of BALF were increased from<1×105 cells/ml in P-P group to>5×105 cells/ml in model group,in which eosinophils were predominant cellular type,levels of IL-4,IL-13,IL-17A in lung and IL-5,IL-13 levels in serum were significantly increased,as well as significant increasing levels of total IgE and MPE-specific IgE,IgG1,IgG2a.Conclusion:MPE-sensitized and challenged mice induces typical eosinophilic asthma featured with elevated eosinophils,as well as secretion of inflammatory factors of type 2 and type 17,IgE,IgG1 and IgG2a subtypes soars to high levels.

Objective To investigate the clinical and radiological features of children with first attack of inflammatory central nervous system disorders and seropositivity to myelin-oligodendrocyte glycoprotein (MOG).Methods The clinical course,cerebrospinal fluid (CSF),MRI studies,MOG status and outcomes were retrospectively analyzed in children with first attack of inflammatory central nervous system disorders and seropositivity to MOG who were hospitalized in Children's Hospital of Fudan University from January 2016 to April 2017.Results Thirteen patients including 8 males and 5 females were included in this study,the ratio of male/female was 1.6∶ 1,and the median age was six years.Ten patients were diagnosed with acute disseminating encephalomyelitis,and three with clinically isolated syndromes.Seven patients had elevated CSF lymphocyte cells,and five patients had elevated CSF protein.All the patients' sera were tested for the anti-MOG IgG,which ranged from 1∶10 to 1∶100 with cell-based assay.MRI results showed that multiple anatomical areas were involved.Twelve patients had brain lesions,in which 10 patients had multiple lobes involved and four had tumefactive demyelinating lesions.The affected anatomical areas included white matters in 11 cases,thalamus/basal ganglias in nine,corpus callosums in three,brainstems in 10,spinal cord in five.The MRI features were characterized by hazy,bilateral lesions without clear boundaries.Clinical symptoms were fully restored in all the patients after treated with intravenous globulin and methyl prednisone.The average follow-up time was 8.9 months,and none of the patients had clinical recurrence.Conclusions MOG was associated with many kinds of inflammatory demyelinating diseases of central nervous system in children.Most of them were diagnosed with acute disseminating encephalomyelitis which has an acute or sub acute clinical course.The clinical manifestations of patients showed diversity.Multiple anatomical areas were involved,and treatment with intravenous globulin and methyl prednisone was effective in the acute phase.All of the patients had a favorable outcome.

Since the definition of sepsis has been first proposed in 1991, the main content of the definition has been developed by infection + systemic inflammatory response syndrome as infection + organ dysfunction,and organ dysfunction is indicated by an increase in sequential organ failure assessment(SOFA) score.These are components of Sepsis-3 which was published in 2016.In Sepsis-3,the SOFA score in the identification and diagnosis of sepsis plays an important role,but Sepsis-3 was mainly used in adults,how to adjust it to children,some questions and details were proposed in this article.

Objective: To summarize the gene mutation of early onset epileptic spasm with unknown reason. Method: In this prospective study, data of patients with early onset epileptic spasm with unknown reason were collected from neurological department of Children's Hospital of Fudan University between March 2016 and December 2016. Patients with known disorders such as infection, metabolic, structural, immunological problems and known genetic mutations were excluded. Patients with genetic disease that can be diagnosed by clinical manifestations and phenotypic characteristics were also excluded. Genetic research methods included nervous system panel containing 1 427 epilepsy genes, whole exome sequencing (WES), analysis of copy number variation (CNV) and karyotype analysis of chromosome. The basic information, phenotypes, genetic results and the antiepileptic treatment of patients were analyzed. Result: Nine of the 17 cases with early onset epileptic spasm were boys and eight were girls. Patients' age at first seizure onset ranged from 1 day after birth to 8 months (median age of 3 months). The first hospital visit age ranged from 1 month to 2 years (median age of 4.5 months). The time of following-up ranged from 8 months to 3 years and 10 months. All the 17 patients had early onset epileptic spasm. Video electroencephalogram was used to monitor the spasm seizure. Five patients had Ohtahara syndrome, 10 had West syndrome, two had unclear classification. In 17 cases, 10 of them had detected pathogenic genes. Nine cases had point mutations, involving SCN2A, ARX, UNC80, KCNQ2, and GABRB3. Except one case of mutations in GABRB3 gene have been reported, all the other cases had new mutations. One patient had deletion mutation in CDKL5 gene. One CNV case had 6q 22.31 5.5MB repeats. Ten cases out of 17 were using 2-3 antiepileptic drugs (AEDs) and the drugs had no effect. Seven cases used adrenocorticotropic hormone (ACTH) and prednisone besides AEDs (a total course for 8 weeks). Among them, five cases had no effect and two cases were seizure free recently. A case with GABRB3 (C.905A>G) had seizure controlled for 3 mouths. A case with ARX (C.700G>A) had seizure controlled for 6 mouths. Conclusion The early onset epileptic spasm with unknown reason is highly related to genetic disorders. A variety of genetic mutations, especially new mutations were found. Genetic heterogeneity of epileptic spasm is obvious.

Objective: To investigate the clinical features and genetic characteristics of patients with TBC1D24 gene mutations. Method: The clinical data of a patient with novel TBC1D24 compound heterozygous mutations from Children′s Hospital of Fudan University were collected, the related literature was searched from China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, National Center from Biotechnology Information and Pubmed (up to April 2016) by using search terms "TBC1D24" "epilepsy" . The clinical features, electroencephalogram (EEG) and prognosis of the patients with TBC1D24 gene mutations were studied. Result: The patient was a boy with non-consanguineous healthy parents.He had an acute episode of focal continuous myoclonus lasting a few hours with consciousness preserved at the age of 3 months.Myoclonic jerks alternatively affected the eyelids, either the right or left limbs, sometimes triggered by fever or fatigue.The frequency was once 3-7 days.At the age of 6 months he was found to have myoclonus seizures with onset from a unilateral eyes lid and limb lasting 10 more minutes and subsequently affected four extremities or the trunk.They occurred once 3-4 months with perserved consciousness and lasted from several hours to up to ten more hours.They mostly disappeared during sleep.He had ataxia and mild mental retarding.Paroxysmal anomalies were not found on ictal traces.A novel compound heterozygous mutation of TBC1D24 gene, c. 730G>A, p.A244T and c. 1571G>C, p.R524P were found in the patient.Further study showed that c. 730G>A mutation was inherited from his father and c. 1571G>C from his mother. These two were not reported in public databases and predicted deleterious by Mutation Taster and polyphen-2.Literature relevant to TBC1D24 published all around the world was reviewed, no Chinese cases with TBC1D24 gene mutations had been reported. The total of 24 cases including the present case with TBC1D24 gene mutation were reported.Among them, 11 cases had compound heterozygous mutations and 13 cases had homozygous mutations.Ten mutations were identified, including 1 termination mutation, 1 frameshift mutation and 8 missense mutations. Conclusion TBC1D24 gene mutational analysis should be performed on patients with early-onset focal continuous myoclonus, if the etiology was unclear.

OBJECTIVE:To provide reference for the improvement of Pharmaceutical Equipment & Workshop Design teach-ing. METHODS:Task-driven teaching was adopted,CIDO engineering educational model was referred to,task was designed from the conception to run with task-based;theoretical and teaching exploration and practice was carried out by cross-learning of individ-uals and groups,applying of pilotscale experiment platform,developing course design and forming evaluation. RESULTS & CON-CLUSIONS:It changed the traditional teaching method that emphasized on class teaching to make students participate in multi-task learning,so that students can better grasp the equipment selection,understanding and application of workshop design in pharmaceu-tical process. The method gives full play to students' initiative and creativity and it is conducive to students’comprehensive ability for adapting the development of pharmaceutical industry.

Objective To investigate the relationship among health literacy and treatment compliance and prognosis in patients with diabetes combined tuberculosis.Methods One hundred and eighty-one cases patients with diabetes combined tuberculosis were selected as the objectives and evaluated literacy level,compliance scale by the health literacy scale and compliance questionnaires.The control of tuberculosis,blood glucose control,treatment compliance and prognosis were analyzed in the patients with different health literacy level.Results Of 181 patients,there were 89 cases patients'(basic health literacy group) the total scores of health literacy and compliance were＞80 points,the efficacy of pulmonary tuberculosis control and blood sugar control were respectively 84.2％ (75/89) and 93.2％ (83/89);92 cases patients' the total scores of health literacy and compliance were ≤ 80 points,the efficacy of pulmonary tuberculosis control and blood sugar control were respectively 59.7％ (55/92) and 67.4％ (62/92),the differences were significant (x2 =12.78,18.11;P ＜0.05).Pearson correlation analysis showed that the total scores of health literacy were positive correlated with the scores of treatment compliance (r =0.688,P ＜ 0.001).Logistic regression analysis showed that three dimension of health literacy (health knowledge and concept,healthy lifestyle and behavior,health skills) that can combined predict the variance of rehabilitation compliance(P＜0.01).Conclusion The health literacy level of diabetes combined tuberculosis patients can effectively predict the rehabilitation compliance.Measures should be taken to improve the patient's level of health literacy,so as improve the compliance of rehabilitation and prognosis.

Objective To investigate the effects of notoginseng and guiding medicinals mediated notoginseng for improving the renal inter stitial fibrosis in rats with chronic kidney-disease(CKD)by regulating TGF-β signaling pathway.Methods A total of 100 male SD rats were randomly divided into five groups:normal group(NOR,n =20),model group(CKD,n =20),radix notogin-seng group(RN,n =20),radix notoginseng plus platycodi group (RNP) and radix notoginseng plus cinnamon group (RNC,n =20).Except for the NOR group,the CKD rat model in other groups was established by adenine gavage.After modeling,the NOR group and CKD group were given the same volume of normal saline by gavage,while the group RN,RNP and RNC were given corresponding drugs by gavage,for 4 weeks.After 4 weeks,the rats in each group were sacrificed for collecting serum and detecting the renal function(serum Scr,BUN),the renal tissues were taken for conducting HE and Masson staining.Then the renal tissue pathological damage severity was observed.The expressions of FN and LN in kidney tissue were detected by immunohistochemistry and the expressions of TGF-[β,α-SMA were detected by Western blot method.Results Compared with the NOR group,the model group exhibited the renal dysfunction(P＜0.01),renal interstitial severe fibrosis manifestation and increased collagen deposition(P＜0.05),and the expression of kidney tissues α-SMA(P＜0.01),TGF-β(P＜0.01),FN and LN were significantly increased.Compared with the model group,the renal function in various treatment groups was improved,Scr(P＜0.01)and BUN(P＜0.01)were significantly decreased,the renal interstitial fibrosis degree was reduced,collagen desposition was decreased(P＜0.05),renal tissue α-SMA(P＜ 0.05),TGF-β(P＜0.05),FN and LN expression were reduced to some extent,in which the effect of RNC group was stronger than that of the RN group and RNP group.Conclusion Notoginseng and guiding medicinals mediated notoginseng can retard the progression of renal interstitial fibrosis caused by adenine in CKD rat in varying degrees,its mechanism maybe reduce the expression of TGF-β protein.

PURPOSE: Cell-in-cell structures are created by one living cell entering another homotypic or heterotypic living cell, which usually leads to the death of the internalized cell, specifically through caspase-dependent cell death (emperitosis) or lysosome-dependent cell death (entosis). Although entosis has attracted great attention, its occurrence is controversial, because one cell line used in its study (MCF-7) is deficient in caspase-3. METHODS: We investigated this issue using MCF-7 and A431 cell lines, which often display cell-in-cell invasion, and have different levels of caspase-3 expression. Cell-in-cell death morphology, microstructures, and signaling pathways were compared in the two cell lines. RESULTS: Our results confirmed that MCF-7 cells are caspase-3 deficient with a partial deletion in the CASP-3 gene. These cells underwent cell death that lacked typical apoptotic properties after staurosporine treatment, whereas caspase-3-sufficient A431 cells displayed typical apoptosis. The presence of caspase-3 was related neither to the lysosome-dependent nor to the caspase-dependent cell-in-cell death pathway. However, the existence of caspase-3 was associated with a switch from lysosome-dependent cell-in-cell death to the apoptotic cell-in-cell death pathway during entosis. Moreover, cellular hypoxia, mitochondrial swelling, release of cytochrome C, and autophagy were observed in internalized cells during entosis. CONCLUSION: The occurrence of caspase-independent entosis is not a cell-specific process. In addition, entosis actually represents a cellular self-repair system, functioning through autophagy, to degrade damaged mitochondria resulting from cellular hypoxia in cell-in-cell structures. However, sustained autophagy-associated signal activation, without reduction in cellular hypoxia, eventually leads to lysosome-dependent intracellular cell death.
Apoptosis ; Autophagy ; Caspase 3* ; Cell Death ; Cell Hypoxia ; Cell Line ; Cytochromes c ; Entosis ; MCF-7 Cells* ; Mitochondria ; Mitochondrial Swelling ; Staurosporine

Objective:To analyse the physicochemical properties and structure of major privet pollen allergen Lig v 1 using bioinformatics software and provide a reference for choosing suitable recombinant expression system for Lig v 1 and modifying the allergen Lig v 1 experimentally.Methods:The physicochemical properties were analysed by ProtParam,the signal peptide by SignalP 4.1 Server,the transmembrane helix by TMHMM Server v.2.0,the secondary structure by GOR4,MHCⅡepitopes by NetMHCⅡ2.2 Server,B-cell epitopes by ProteanTM 5.01 and the phylogenetic tree by MEGA 6.Results: Privet major pollen allergen Lig v 1 was stable in Escherichia coli and it doesn′t possess any signal peptide and transmembrane helix.Most secondary structures of Lig v 1 were random coils.Potential region of MHCⅡepitope of Lig v 1 was 30-44.Potential B-cell epitopes possess discontinuous and continuous a mino acid sequences.Lig v 1 and its counterparts from Fraxinus excelsior and Olea europaea were clustered into one group.Conclusion:Escherichia coli is the suitable expression system for recombinant Lig v 1.In silico prediction of the epitopes of Lig v 1 provides a reference for modifying the allergen Lig v 1 experimentally.