Objective:To investigate the effect of liquiritin on the apoptosis of amygdala cell and the expression of apoptosis-related factors Bax and Bcl-2 protein in rats with post-stroke depression (PSD).Methods:Sixty rats were randomly divided into normal control group, stroke group, PSD group, citalopram group, liquiritin group, and normal saline control group ( n=10 in each group). The middle cerebral artery was occluded with a suture method to induce focal cerebral ischemia, and the PSD model was established by chronic and unpredictable mild stress stimulation and orphanism. At the same time every week after the model was made, the weight of rats in each group was measured and the depression behavior was evaluated, including sucrose water test and open field test. At 6 weeks after the model was made, TUNEL staining was used to detect the apoptosis of amygdala cell, immunofluorescence staining was used to detect the expression of Bax and Bcl-2 in the amygdala, and Western blot analysis was used to detect the protein expression of Bax and Bcl-2 in the amygdala. Results:Compared with the liquiritin group, citalopram group and normal control group, the body weight and sucrose solution preference of rats in the stroke group, PSD group and normal saline control group were decreased, and the horizontal and vertical movements in open field test were decreased; the differences were statistically significant (all P<0.01). TUNEL staining results showed that compared with the liquiritin group, citalopram group and normal control group, the number of apoptotic cells was significantly increased in the stroke group, PSD group, and normal saline control group; the difference was statistically significant (all P<0.01). The results of immunofluorescence staining showed that compared with the liquiritin group, citalopram group and normal control group, the number of bcl-2 immunoreactive cells in amygdala of the stroke group, PSD group and normal saline control group was significantly decreased, while the number of Bax immunoreactive cells was significantly increased; the difference was statistically significant (all P<0.01). Western blot analysis showed that compared with the liquiritin group and citalopram group, the expression of bcl 2 protein in amygdala of the stroke group, PSD group and normal saline control group was significantly decreased, while the expression of Bax protein was significantly increased; the difference was statistically significant (all P<0.01). Conclusion:Liquiritin can alleviate the symptoms of PSD, and its mechanism may be related to inhibiting the apoptosis of amygdala cells and regulating the expression of apoptosis-related factors.

Objective:To observe the changes of protein expression of apoptosis signal pathway in prefrontal cortex of rats with post-stroke depression(PSD) after lateral ventricle injected of brain-derived neurotrophic factor precursor(proBDNF).Methods:Among 55 healthy adult female SD rats, 25 rats were randomly selected as PSD group, and the other 30 rats were randomly divided into normal group ( n=10), depression group ( n=10) and stroke group ( n=10). The middle cerebral artery occlusion(MCAO) model was established by thread occlusion in the stroke group, the chronic stress depression model in the depression group was established by the combination of chronic unpredictable mild stress(CUMS) and the solitary feeding method.And the rats in the PSD group were established MCAO model first, then they were received CUMS stress and solitary rearing one week later so as to establish PSD model.Two weeks after the establishment of the model, 15 rats in PSD group were randomly divided into proBDNF group, rats in tPA group and NS control group.One week after buried tube of lateral ventricle, rats in tPA and proBDNF were injected into the lateral ventricle for one week.The protein expressions of c-Jun N-terminal kinase(JNK), p-JNK, p53, p-p53 and Bax in prefrontal cortex of rats in each group were detected by Western blot at the 4th and 8th week after modeling.SPSS 17.0 software was used for data analysis, one-way ANOVA was used for comparison between groups, and SNK- q was used for pairwise comparison. Results:The expressions of p-p53, p53, p-JNK, JNK and Bax in prefrontal cortex of normal group, depression group, stroke group and PSD group were significantly different at the end of 4th and 8th week after MCAO modeling ( F=3.426-90.355, all P<0.05). Post-hoc analysis showed that, compared with the normal group, the expressions of p-JNK (0.378±0.042) and Bax (0.478±0.054) in the prefrontal cortex of PSD rats increased significantly at the end of the 4th week(both P<0.05), and the expressions of p-JNK(0.411±0.056), p-p53 (0.286±0.083) and Bax (0.471±0.008) in the prefrontal cortex of PSD group increased significantly at the end of the 8th week(all P<0.05). After lateral ventricle injection of proBDNF, there were significant differences in the expression of p-p53, p53, p-JNK, JNK and Bax among proBDNF group, tPA group and NS group ( F=16.915-287.039, all P<0.01). Post-hoc analysis showed that, compared with NS group, the expressions of p-JNK (0.35±0.01)and p-p53 (0.31±0.01)in prefrontal cortex of proBDNF group increased significantly(both P<0.05). After lateral ventricle injection of proBDNF, there were significant differences in body weight, sucrose preference rate, horizontal movement distance among proBDNF group, tPA group and NS group ( F=18.741-76.305, all P<0.01), and compared with tPA group and NS group, behavioral indexes of proBDNF group (body weight (224.36±3.23) g, sucrose preference rate (69.83±1.72)%, horizontal movement distance (57.93±2.09) blocks, vertical movement distance (19.79±1.81)) decreased significantly(all P<0.05). Conclusion:The proBDNF promotes the activation of apoptosis signal pathway in the rats with PSD.

To investigate the upregulated genes associated with cold acclimation, a cold acclimation model was established based on Balb/C mouse. mRNA of muscle and liver were isolated, and the upregulated genes of these tissues were studied by representational differential analysis (RDA). The upregulated genes then were sequenced and searched by Blast software in GenBank database. The results showed that some genes were upregulated and possibly associated with cold acclimation. Three of these genes, transferrin, fibrinogen B-beta-chains and a new gene fragment (Genbank ID: AF454762), were confirmed to be upregulated by RNA slot-blot analysis. The finding of these genes might contribute to further understanding of the molecular mechanisms of cold acclimation.
Acclimatization ; genetics ; Animals ; Cold Temperature ; Gene Expression ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Muscle, Skeletal ; metabolism ; Transcriptome ; Up-Regulation

Firstly, IHE technical framework is introduced in this paper, and the sub-framework of Schedule Workflow is also described. Secondly, the development of IHE in Japan and the formation of IHE-J are presented, and in combination with IDS system's practice in IHE-J, some ideas about developing IHE in China and the relative methods of implementation are discussed.
China ; Computer Communication Networks ; Hospital Information Systems ; Japan ; Radiology Information Systems ; organization & administration ; Software ; Systems Integration ; User-Computer Interface ; Workload

To investigate JAK2V617F mutation and its clinical significance in patients with idiopathic myelofibrosis (IMF), genomic DNA was extracted from peripheral blood cell samples of 12 IMF cases. Allele-specific PCR (AS-PCR) was performed to identify JAK2V617F mutation, and the results were confirmed by sequence analysis. A retrospective study was performed to explore the correlation between JAK2V617F mutation and the clinical, hematologic features. The results showed that in follow-up for 2 to 15 months, the occurrence of the positive point mutation in 12 patients with IMF was 50%, and the half of these positive patients had thrombosis. Patients with JAK2V617F point mutation had a higher counts of platelets and megakaryocytes in bone marrow than those in patients without JAK2V617F point mutation. Out of other 6 IMF patients without JAK2V617F point mutation only 1 patient had thrombosis, and lower counts of platelets in peripheral blood and megakaryocytes in bone marrow. It is concluded that majority of IMF patients with positive JAK2V617F point mutation have typical clinical and hematologic features, higher incidence of thrombosis, and higher counts of platelets in peripheral blood and megakaryocytes in bone marrow.
Adult ; Aged ; Base Sequence ; Bone Marrow ; pathology ; Female ; Follow-Up Studies ; Humans ; Janus Kinase 2 ; genetics ; Male ; Megakaryocytes ; pathology ; Middle Aged ; Molecular Sequence Data ; Platelet Count ; Point Mutation ; Primary Myelofibrosis ; genetics ; Retrospective Studies

OBJECTIVETo study the anti-tumor effect of Biejiajianwan pills and explore its mechanism.

METHODSMice bearing neoplasm induced by H22 cell inoculation were randomized into 4 groups, namely the negative control group, positive control group (with cyclophosphamide treatment), and high- and low-dose Biejiajianwan pill group. The tumor-bearing mice received corresponding treatments for 15 consecutive days, after which their thymuses and spleens were isolated and weighed to calculate the thymus and spleen indices. The tumor mass was also weighed and the tumor inhibition rate calculated. Immunohistochemistry was performed to evaluate the expression intensity of proliferating cell nuclear antigen (PCNA) in the tumor tissue.

RESULTSThe pills significantly inhibited the growth of the implanted tumor and reduced the expression of PCNA in the tumor.

CONCLUSIONOne of the possible mechanisms of the anti-tumor effects of Biejiajian pills might be associated with the inhibition of PCNA expression in the tumor.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; therapeutic use ; Carcinoma, Hepatocellular ; metabolism ; pathology ; prevention & control ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Immunohistochemistry ; Liver Neoplasms, Experimental ; metabolism ; pathology ; prevention & control ; Male ; Mice ; Proliferating Cell Nuclear Antigen ; biosynthesis ; Random Allocation ; Tumor Burden ; drug effects

Thirst study was purposed to explore the genetic polymorphism of Chinese Zhuang population at HLA-Cw locus by sequence based typing (SBT). A total of 150 unrelated blood samples from Chinese Zhuang population were subjected to sequencing at exon 2, 3 and 4 of HLA-Cw gene in both directions by using SBT technique established by our laboratory. The purified products of sequencing reaction were run by means of electrophoresis on the ABI 3730 DNA Sequencer and the assignment of HLA-Cw genotype was accomplished by using the Assign 3.5 software. The consensus sequence at exon 2, 3 and 4 of HLA-Cw gene for each sample was imported into the Assign 3.5 software. The results showed that 33.33% of tested samples could obtain an unique genotype, genotype in 63.33% of tested samples with ambiguous results could be assigned by ruling out the rare alleles according to the NMDP Rare Allele List File; however, the final genotype in rest 3.33% of the detected samples could be defined when subjected to further confirmatory testing by PCR-SSP. In this detection 16 HLA-Cw alleles were identified, the common alleles with a frequency of > 10% were Cw*0304 > Cw*0102 > Cw*0801 > Cw*0702. The value for gene diversity (GD) was 0.9297, The frequency for Cw*01, 03, 07, 08, 12, 14 (Cw 1 allele group) and Cw*02, 04, 05, 06, 15, 16, 17, 18 (Cw 2 allele group) was 0.8967 and 0.1032, respectively, which indicated that the Cw 1 allele group is the dominant ligand for KIR in Chinese Zhuang population. 51 genotypes were determined and the distribution of genotype frequency was in line with Hardy-Weinberg principle. It is concluded that the obtained HLA-Cw allele frequency and its distribution characteristics of Chinese Zhuang population can provide valuable data in the studies of anthropology and the association of HLA-Cw with disease.
Asian Continental Ancestry Group ; genetics ; Exons ; Gene Frequency ; Genotype ; HLA-C Antigens ; genetics ; Humans ; Molecular Sequence Data ; Polymorphism, Genetic ; Sequence Analysis, DNA

BACKGROUNDTo clarify the features of gene variation among epidemic strains of human rotavirus NSP4 in China.

METHODSSP4 genes from 27 epidemic strains of human rotavirus isolated in different area of China in recent years were amplified with RT-PCR, the resulted cDNAs were cloned and sequenced. The sequences of full length cDNAs were compared with 10 rotavirus NSP4 sequences available in the GenBank using the Clustal x 1.8 TreeView32 and DNA Star softwares. The G serotype of VP7 was analysed by PCR.

RESULTSThe homology of the amino acid among the 27 rotavirus strains isolated in China was 81.7%-99.4%. Based on the variation of amino acid sequence, the virus strains can be divided into two groups, represented by Wa and KUN with the homology of 92.0%-99.4% and 92.0%-98.9% within each group, respectively. The diversity between the two groups were 16.6%-21.0%. The Wa group could further be separated into three subgroups, according to the diversity between those strains and the characterization in the highly variable domain. The association between VP7 serotype and NSP4 genotype was not strong.

CONCLUSIONSThe NSP4 gene of human rotavirus epidemic strains in China can be divided into Wa and KUN two groups, Wa group is the main group and contains three subgroups possessing characteristic amino acid sites. Samples isolated in the same year but not in the same area shared higher homology.

Antigens, Viral ; Capsid Proteins ; genetics ; DNA, Complementary ; analysis ; DNA, Viral ; analysis ; Diarrhea ; virology ; Genetic Variation ; Genotype ; Glycoproteins ; genetics ; Humans ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; genetics ; isolation & purification ; Sequence Analysis, DNA ; Toxins, Biological ; Viral Nonstructural Proteins ; genetics

OBJECTIVETo explore the relationship between the proliferation, differentiation of rat hepatic stem like cell line WB-F344 and cytokines in vitro.

METHODS(3)H thymidine labelling of new synthesized DNA was used to examine the mitogenic responsiveness of WB-F344 cells to cytokines, western blot was used to study the expression of cytokines receptors on hepatic stem cells, and apoptotic cells were detected by Flow cytometry.

RESULTSWB-F344 cells showed a proliferative response to the cytokines of hepatocyte growth factor (HGF), epidermal growth factor (EGF), fibroblast growth factor (FGF), Insulin at the dose of 80 ng/ml, and the relative cpm values are 982.95, 906.32, 863.98 and 968.67 respectively, while non response to interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) at the same dose, and an inhibition or apoptosis response to transforming growth factor-beta (TGF-beta) at 80 ng/ml with a 26.89% apoptotic rate. Western blot showed that there were HGF, EGF, FGF, TGF-beta receptors expressed on WB-F344 cells. When WB-F344 cells were cultured in the differential system (DMEM, 10% Fcs, HGF 10 to approximately 50 ng/ml, EGF 20 ng/ml, Insulin 1 microg/ml, Dex 1 micromol/L), the cells could differentiated into hepatocytes. In addition, HGF could scattered WB-F344 cells.

CONCLUSIONThe proliferation and differentiation of liver stem cells are regulated by various cytokines which may play an important role when liver is damaged seriously.

Animals ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; Cytokines ; pharmacology ; Epidermal Growth Factor ; pharmacology ; Hepatocyte Growth Factor ; pharmacology ; Insulin ; pharmacology ; Liver ; cytology ; Rats ; Rats, Inbred F344 ; Stem Cells ; cytology ; drug effects ; Transforming Growth Factor beta ; pharmacology

Objective:To use polyamidoamine(PAMAM)dendrimer as gene delivery system for survivin gene anti- sense oligodeoxynucleotide(asODN)transfection for inhibition of HepG2 cancer cell growth.Methods:The first to the fifth generation of PAMAM and asODN were used to prepare a complex:PAMAM-asODN.The morphology of PAMAM- asODN was observed using agrose electrophoresis and atomic force microscope(AFM).PAMAM-asODN was then used to transfect HepG2 cells and cells transfected with asODN served as control.The transfection efficacy of PAMAM-asODN into HepG2 cells was observed under confocol microscope,the surviving mRNA expression was analyzed by RT-PCR,and the inhibition of HepG2 cell growth was determined by MTT assay.Results:Agrose electrophoresis showed strong complexing action between PAMAM and asODN and they formed a complex with a diameter of 25 nm.Confocol microscope showed the transfection efficacy of PAMAM-asODN was higher than that of asODN.RT-PCR showed a decreased expression of sur- vivin mRNA in PAMAM-asODN transfected cells.MTF results demonstrated that the growth of HepG2 cell was obviously inhibited after transfection of PAMAM-asODN and the inhibition rate increased with culture time,concentration of com- plex,the generation of PAMAM.PAMAM-asODN at 6.0?mol/L G4.0 resulted in a 55% inhibition of HepG2 cells 96 h after culture.Conclusion:PAMAM dendrimers can efficiently mediate the entry of survivin asODN into HepG2 cells,re- sulting in inhibition of HepG2 cells.PAMAM might be a promising gene carrier for potential molecular therapy of cancer.