ObjectiveTo investigate the protective effects of epigallocatechin gallate (EGCG) in a mouse model of Parkinson's disease induced by 1-Methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP).Methods32 C57BL/ 6 male mice were randomly divided into 4 groups: Model group was administrated with 16 mg/kg MPTP (i.p., four times, 2 h interval); Sham group was treated with saline; EGCG treatment group was given EGCG (5 mg/kg) after MPTP administration; normal group was just given EGCG (5 mg/kg) as treatment group. After given EGCG for 3 weeks, behavioral tests, tyrosine hydroxylase (TH) immunohistochemistry staining and the HPLC for dopamine (DA) and its metabolites were used.ResultsThe present results indicated that oral administration of EGCG significantly improved the behavioral impairement in mice induced by MPTP (P<0.05). And in the EGCG treatment group, there were more TH-positive neurons than in model group. In addition, levels of DA and its metabolites in striatum decreased significantly in MPTP group (P<0.05). Though the concentration of DA and its metabolites in EGCG treatment group tended to increase, however, there was no significance between EGCG treatment and model group.ConclusionEGCG could improve the behavioral impairment in a mouse model of Parkinson's disease induced by MPTP and protect against the loss of the dopamine neurons in the substantia nigra (SN).

BACKGROUND: Changes in extracellular of chondrocyte can reflect influence of external force on temporomandibular joint and adaptability of body to external force. OBJECTIVE: To study the effect of cyclic tensile stress on main extracellular matrix of condylar chondrocyte.METHODS: The cyclic tensile stress was exposed to the third passage condylar chondrocyte for 0, 1, 6, 12 and 24 hours, respectively, using a Flexcell Strain Unit-5000T system (10% surface elongation, 6 cycles/min). After mechanical loading, total RNA was extracted from the cells harvested from Six-well BioFlex flexible cell Petri Dish, reverse transcribed, and reverse trabscription-PCR was performed to quantify mRNA levels for type Ⅱ collagen and aggrecan.RESULTS AND CONCLUSION: Compared with the control group (0 hour group), both type Ⅱ collagen and aggrecan mRNA expression was significantly increased after loading for 6 hours (P < 0.05), but began to decrease since 12 hours, and significantly decreased at 24 hours (P < 0.05). Results showed that cyclical tensile stress stimuli can affect the synthesis of main extracellular matrix of condylar cartilage, i.e. the synthesis was gradually enhanced with prolonged stimulation duration, but significantly inhibited in response to further stress stimuli.

Strict regulation of HIV-1 PR function is critical for efficient production of mature viral particles. During viral protein expression and viral assembly, HIV-1 PR located within Gag-Pol precursor must be inactive to prevent premature cytoplasmic processing of the viral Gag and Gag-Pol precursors. Premature activation of HIV-1 precursors leads to major defects in viral assembly and production of viral particles. A cell-level premature activation of HIV-1 precursors assay using bioluminescence resonance energy transfer (BRET) was established. Three thousand compounds were screened to evaluate this assay. The results showed that the assay is sensitive, specific and stable (Z' factor is 0.905).

The HIV-1 Rev protein facilitates nuclear export of unspliced and singly spliced viral transcripts containing RRE RNA through the CRM1 export pathway. Inhibition of Rev-mediated RNA nuclear export can arrest HIV-1 transcriptional process, which clearly, reveals a target for anti-HIV drug development. In this work, a cell-based assay has been established for screening anti-HIV compounds targeting the Rev-mediated RNA nuclear export. This assay utilized a codon-optimized green fluorescent protein (GFP) as reporter gene, which expression is in a Rev-dependent manner. Any compound that inhibits the Rev-mediated RNA nuclear export is identified by reducing emission of GFP. The Z' score of this model is 0.8220. Three thousands compounds were screened and the positive rate was 9.3% with a cutoff at 50% inhibition. IMB7C7, one of the positive compounds, efficiently inhibits viral production from HIV-1 infected cells.

Objective Establishing a fingerprint method to identify the characteristic chemicals in the roots of Gentiana macrophylla and evaluate their quality.Methods RP-HPLC was developed for fingerprint analysis and determination of four ingredients in G macrophylla roots from different sources.LC-ESI-TOF-MS was employed to identify the chromatographic peaks of the fingerprint.Results Five common peaks were identified by comparing their retention time with reference secoiridoid glucosides.Eight major peaks in chromatographic fingerprint were analyzed by on-line LC-ESI-TOF-MS.Four secoiridoid glucosides were identified based on their MS data.Conclusion The method is specific and could be served for the quality identification and comprehensive evaluation of G macrophylla.

Objective To develop Community Home Mutual Pension Demand Scale and test its validity and reliability.Methods Firstly,the theoretical dimension of Community Home Mutual Pension Demand Scale was constructed by means of literature analysis and resident interview.Secondly,the scale with five dimensions containing 29 items was formed after small range of questionnaire survey.Finally,the data from 384 elderly people in 4 communities of Hengyang city were analyzed by using SPSS17.0 and AMOS17.0 to test its validity and reliability.Results The model consisted of 4 dimensions and 21 items,which explained 69.911％ of the total variance.x2/df=2.097,Tac ker Lewis index (TLI)=0.959,non normed fit index (NNFI)=0.959,relative fit index (RFI)=0.924,goodness-of-fit index(GFI)=0.921,comparative fit index (CFI)=0.966,normed fit index (NFI)=0.938,incremental fit index (IFI)=0.967,root-mean square error of approximation(RMSEA)=0.054;Cronbach α of the total scale was 0.932,Cronbach α of each dimension was 0.801-0.954.The correlation coefficient between dimensions was 0.241-0.592,P＜0.01.Conclusions The scale has good reliability and validity in the measurement of the mutual pension demand of elderly.

With the emergence of drug resistant tuberculosis, it is very urgent to find novel anti-tuberculosis drugs, especially novel anti-drug-resistant tuberculosis drugs. Because of the slow growth and the need to work in a biosafty environment of Mycobacterium tuberculosis, the development of evaluation of drug effect is severely impeded. In order to solve these issues, non-pathogenic fast-growing Mycobacterium smegmatis is introduced as test organism. The inhA is one of a target of isoniazid (INH) overexpression or mutation of this gene in Mycobacterium tuberculosis conferring resistant to INH. A recombinant plasmid bearing inhA was constructed and electroporated into Mycobacterium smegmatis, using shuttle expression vector pMV261. Transformants were induced to express a protein of inhA, identified by SDS-PAGE. Results show that Mycobacterium smegmatis containing inhA plasmids exhibited 100-fold or greater increased resistance to INH, but it conferred no increased resistance to others first-line anti-tuberculosis drugs. Resazurin microtiter assay plate testing of Mycobacterium smegmatis susceptibility to drugs is a rapid, simple, and inexpensive method and could decrease color background of drugs by detecting fluorescence. It will be benefit for high-throughout screening of drugs of anti-isoniazid-resistant Mycobacteria.

OBJECTIVE: To report on a family which has two siblings with SCN2A mutation caused by germline mosaicism suffering from autism spectrum disorder/development delay (ASD/DD). METHODS: Clinical data was collected for the proband and his parents. Next generation sequencing (NGS) was carried out on the proband and his parents. Suspected mutations were verified by Sanger sequencing of the proband, his parents and brother. To detect whether there is a low proportion of somatic mosaicism in the parents, a droplet digital PCR was conducted. The result of ddPCR showed that the father was germline mosaicism (0.233%). RESULTS: NGS has identified a de novo splicing mutation of the SCN2A gene, c.605+1G>A, in the proband and his brother. Combined with its clinical phenotype and inheritance pattern, SCN2A was judged to be the pathogenic gene. Above findings strongly suggested parental germline mosaicism. CONCLUSION ASD/DD in siblings with SCN2A mutations caused by germline mosaicism. Paternal mosaicism should be considered as one of the important inheritance patterns for counseling parents with a child carrying SCN2A mutation. The ddPCR can help to reveal very low proportion of germline mosaicism.
Autism Spectrum Disorder ; Germ Cells ; Humans ; Male ; Mosaicism ; Mutation ; NAV1.2 Voltage-Gated Sodium Channel/genetics* ; Siblings

A case of IgG4-related hypertrophic pachymeningitis was reported. The patient was an elderly female, with the course of disease more than 8 years. Clinical manifestations included recurrent headache, vision and hearing loss, exophthalmos and thyroid dysfunction. Finally, she was diagnosed as IgG4-related disease and IgG4-related hypertrophic pachymeningitis by PET-CT and dural biopsy. After treatment with methylprednisolone and mycophenolate mofetil, the patient′s clinical symptoms improved.

Objective:To explore the effects of Jianpi Bushen Jiedu Prescription on the proliferation and migration of hepatocellular carcinoma cells; To discuss its possible mechanism.Methods:Using human highly metastatic liver cancer cell line (HCCLM3) as the research object, they were randomly divided into control group and TCM group (100, 200, 400, 800, 1 600, 3 200 μg/ml Jianpi Bushen Jiedu Prescription) and Western medicine group (2.5, 5, 10, 20, 40 μmol/L sorafenib) using a random number table method. Cell viability was detected using cell counting reagent (CCK-8) method; HCCLM3 cells were divided into control group and TCM (Jianpi Bushen Jiedu Prescription 800 μg/ml) group and combined group (Jianpi Bushen Jiedu Prescription 800 μg/ml +sorafenib 20 μmol/L). Western blot method was used to detect the protein expressions of kinase/signaling transducer and transcriptional activator (JAK2/STAT3) pathway related proteins (p-JAK2, JAK2, p-STAT3, STAT3) in each group.Results:Compared with the control group, viability and mobility of HCCLM cell in TCM group and Western medicine group decreased ( P<0.01 or P<0.05); compared with the control group, the protein expressions of P-JAK2, JAK2, P-STAT3 and STAT3 in the TCM group and the combined group decreased ( P<0.05), and the JAK2 protein expression in the combined group was lower than that in the TCM group ( P<0.05). Conclusion:Jianpi Bushen Jiedu Prescription can inhibit the proliferation and migration of HCC cells by regulating JAK2/STAT3 pathway.