Animals ; Conditioning (Psychology) ; drug effects ; Extinction, Psychological ; drug effects ; physiology ; Fear ; drug effects ; physiology ; Male ; Mifepristone ; pharmacology ; Rats ; Rats, Sprague-Dawley

Animals ; Extinction, Psychological ; physiology ; Fear ; physiology ; Male ; Mental Recall ; physiology ; Rats ; Rats, Sprague-Dawley ; Sleep Deprivation ; complications ; physiopathology ; Sleep, REM ; physiology

To observe the effect of Ligusticum wallichii-containing serum on the expressions of Toll-like receptor 4 and myeloid differentiation factor 88 in hepatic stellate cells. Clean-grade SD rats were randomly divided into 5 groups and orally given L. wallichii decoction, colchicine and normal saline for 7 d to prepare L. wallichii-containing serums. Except for the blank group, all of the remaining groups were stimulated with LPS 1 mg x L(-1) for 24 h. After being intervened, the L. wallichii-containing serums were cultured in 5% CO2 incubator at 37 degrees C for 24 hours. The expression of TLR4 and MyD88 were detected by RT-PCR and Western blot. After HSC was stimulated with LPS, TLR4 and MyD88 mRNA and protein expressions were significantly higher than the blank control group (P < 0.01). After being intervened with L. wallichii-containing serum, TLR4 and MyD88 mRNA and protein expressions were notably lower than the model group (P < 0.05 or P < 0.01). In conclusion, L. wallichii-containing serum could regulate the TLR4 signaling pathway and show the anti-fibrosis effect by inhibiting the expression of TLR4 and MyD88 in LPS-induced HSCs.
Animals ; Female ; Hepatic Stellate Cells ; drug effects ; metabolism ; Ligusticum ; Lipopolysaccharides ; pharmacology ; Liver Cirrhosis, Experimental ; drug therapy ; Myeloid Differentiation Factor 88 ; genetics ; physiology ; Phytotherapy ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 4 ; genetics ; physiology

To analyze influenza pathogen spectrum in Yunnan province during 2009-2014 years, and analyze HA and NA genes of influenza A H1N1. Analysis was made on the monitoring date of influenza cases in Yunnan province in recent 6 years, 23 strains of influenza virus of HA and NA gene was sequenced and analyzed by MEGA 5 software to construct phylogenetic tree. 4 times of influenza AH1N1 epidemic peak were monitored from 2009-2014 years in Yunnan Province, as the nucleic acid detection results of influenza A H1N1 accounted for 28.8% of the total. The sequencing result showed that HA and NA gene were divided into 3 groups, one was detected with H275Y mutation strains. Influenza A H1N1 is one of the important subtypes in Yunnan province and their genes have divided into three branches during the period of 2009-2014 years, the vast majority of influenza a H1N1 are still sensitive to neuraminidase inhibitors.
China ; epidemiology ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; metabolism ; Humans ; Influenza A Virus, H1N1 Subtype ; classification ; enzymology ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Molecular Sequence Data ; Mutation ; Neuraminidase ; genetics ; metabolism ; Phylogeny ; Viral Proteins ; genetics ; metabolism

Objective To explore the application of "Work-Study Combination"(referred to "Institutions Join Forces with Hospitals" in this paper) in nursing education system of Shanghai Institute of Health Sciences. Methods Data of teaching plan and curriculum of students from 2005 and 2006 grades were compared.With construction of "National Model of Higher Vocational Institutions",transformation in professional curriculum reforming,teaching resources sharing,teachers staff constructing,in-school and out-school vocational training bases establishing,and students evaluation system were compared.Results Regulated and updated teaching contents and new curriculum reflected vocational education characteristics.The ratios of in-school vocational training,clinical teaching and practice in curriculum rose rapidly.Conclusion The results showed that "Institutions Join Forces with Hospitals" applying in higher nursing vocational education system was a direct expression of combination of work and study to the process of personnel cultivating.

Animals ; Cell Culture Techniques ; Cells, Cultured ; Male ; Neural Stem Cells ; cytology ; Neurons ; cytology ; Olfactory Mucosa ; cytology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of Hedgehog-Gli1 signaling pathway on proliferation, apoptosis and activation of hepatic stellate cells (HSCs) in vitro.

METHODSThe expression of Shh, Smo, Ptc and Gli-1 in HSC-T6 cells was analyzed by RT-PCR. HSC-T6 cells were incubated with various concentration of cyclopamine (0, 50, 100, 150, 200, 250 mumol/L) for 24 hours, cell viability was checked by MTT colorimetric assay, cell cycle was analyzed by flow cytometry, apoptosis was assayed by agarose electrophoresis of DNA and PI-Annexin V fluorescent staining, and the mRNA levels of Gli-1, TGF beta 1, PDGF and Bcl-2 were quantified by real-time RT-PCR.

RESULTSRT-PCR indicated that the components of the Hedgehog-Gli1 signaling pathway were expressed in HSC-T6 cells. MTT assay indicated that cyclopamine inhibited cell viability in a concentration dependant manner (F = 636.81, P less than 0.01). Flow cytometry indicated that cells were piled up at G0/G1 phase in cyclopamine treated cells (65.08%+/-1.50%) as compared to control cells (55.41%+/-2.54%, t = -8.05, P less than 0.01). Cyclopamine treatment resulted in apoptosis as indicated by DNA fragmentation and PI-Annexin V staining. The mRNA levels of Gli-1, TGF beta 1, PDGF and Bcl-2 in cyclopamine treated cells were significantly lower than that in control cells (P less than 0.01).

CONCLUSIONCyclopamine may inhibit the Hedgehog-Gli1 signaling, and hence repress proliferation and promote apoptosis in hepatic stellate cells.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Hedgehog Proteins ; antagonists & inhibitors ; genetics ; metabolism ; Hepatic Stellate Cells ; drug effects ; metabolism ; Humans ; Platelet-Derived Growth Factor ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; Transcription Factors ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Veratrum Alkaloids ; pharmacology ; Zinc Finger Protein GLI1

Overexpression of procollagen gene can cause the extraordinary increase of collagen's synthesis and therefore lead to the keloid and hypertrophic scar. To utilize ribozyme to suppress the expression of procollagen genes, a eukaryotic expression recombinant plasmid containing a dual-ribozyme gene against alpha 1 (I) and alpha 1 (III) procollagen genes was constructed. The ribozyme from in vitro transcription was incubated with target transcripts from recombinant plasmids which separately contained the fragments of the second exons of pro alpha 1 (I) and pro alpha 1 (III) collagen genes under various experimental conditions. The results showed that the dual-ribozyme could efficiently catalyze the specific cleavage of the target RNAs at 37 degrees C, 42 degrees C, 50 degrees C and Mg2+ concentration from 10 mmol/L to 20 mmol/L. This work provided a basis for further study on the ribozyme to suppress the expression of procollagen genes and control the cicatrization.
Base Sequence ; Exons ; Molecular Sequence Data ; Procollagen ; genetics ; RNA ; metabolism ; RNA, Catalytic ; genetics ; metabolism ; Temperature

OBJECTIVETo observe improved effects of Jingjin acupuncture on fine activity of hemiplegic hand in recovery period of stroke.

METHODSFifty cases were randomly divided into an observation group and a control group, 25 cases in each one. Regular western medicine treatment, rehabilitation training and regular acupuncture (in which Shuigou (GV 26), Baihui (GV 20), Neiguan (PC 6), etc. were selected) were applied in both groups. Additionally, muscles in palm side of affected hand, dorsal metacarpophalangeal joints and proximal interphalangeal joints were treated with acupuncture in the observation group, once every other day and electroacupuncture was applied when arrival of qi was acquired. Baxie (EX-UE 9) in the affected hand were needled in the control group, and electroacupuncture was added when arrival of qi was acquired. Ten days of treatment was considered a treatment course, and after two courses Lindmark score, Brunnstrom movement function grade, joint range of hand and Barthel index (BI) were observed in two groups.

RESULTSCompared before the treatment, the Lindmark score in two groups were both improved after the treatment (both P < 0.01). Compared with the control group, the motor coordination ability, sensory function and total score of Lindmark in observation group were obviously improved (differences before and after treatment: 8.24 +/- 3.07 vs 6.84 +/- 2.43, 3.52 +/- 2.33 vs 2.16 +/- 2.12, 11.76 +/- 3.55 vs 9.00 +/- 3.62, all P < 0.05). The Brunnstrom movement function grade was significantly improved in both groups after treatment (both P < 0.01), which was more obvious in the observation group (P < 0.05). The joint range of hemiplegic hand was improved in both groups after treatment (both P < 0.01), which was more obvious in the observation group [differences before and after treatment: (25.35 +/- 10.91) degrees vs (18.65 +/- 7.86) degrees, p < 0.05]. The score of BI was also significantly improved after treatment in two groups (both P < 0.01).

CONCLUSIONThe Jingjin acupuncture could effectively improve fine activity of hemiplegic hand in recovery period of stroke prove daily life ability.

Acupuncture Therapy ; Aged ; Female ; Hand ; physiopathology ; Hemiplegia ; etiology ; physiopathology ; therapy ; Humans ; Male ; Middle Aged ; Movement ; Recovery of Function ; Stroke ; complications ; physiopathology

OBJECTIVETo construct the recombinant virus-like particles containing HCV envelope glycoprotein E1E2 based on sindbis virus vector.

METHODSThe gene encoding HCV envelope glycoprotein E1E2 was cloned into sindbis virus vector to construct recombinant plasmids pBR-XJE1E2 and pVA-XJE1E2, and transfect them into BHK-21 cells to obtain recombinant virus-like particles. The expression of E1 and E2 protein were verified by Western Blot and indirect immunofluorescent assay (IFA).

RESULTSThe results of restriction enzyme digestion, PCR and sequencing analysis showed that the recombinant plasmids were constructed successfully. And the results of RT-PCR, Western blotting and IFA detection showed that the transfect cells could package HCV-like particles of expressing structural proteins E1E2.

CONCLUSIONThe recombinant expression plasmids pBR-XJE1E2 and pVA-XJE1E2 based on sindbis virus vector could package HCV-like particles in eukaryotic cell, which provides a foundation for further study of its in vivo animal immune response.

Animals ; Cells, Cultured ; Cricetinae ; Genetic Vectors ; Hepacivirus ; genetics ; Plasmids ; Recombination, Genetic ; Sindbis Virus ; genetics ; Viral Envelope Proteins ; genetics