Objective To establish the quality standard for Complex Prescription Kudouzi Capsules.Methods TLC was employed to identify Crataegus pinnatifida Bge.、Cassia obtusifolia L.and sophora alopecuroides L..The content of chrysophanol in Cassia obtusifolia L.was determined by HPLC.Results Spots obtained from the test solutions had the same color in reference solution and medical material in the same location,and the blank solution had no interference.The linear range of chrysophanol was 0.028 4 ～ 0.908 8 ?g,r =0.999 7(n =6),and the average recovery was 99.97%.Conclusion The characteristic of identification by TLC was highly specific,HPLC is accurate and reproducible,and they can be used effectively for the quality control of Complex Prescription Kudouzi Capsules.

Objective:To explore characteristics of family function in patients with major depressive disorder (MDD),and to evaluate relationships between family function and prognosis of major depressive episode (MDE).Methods:Forty-six patients with MDD were recruited in the outpatient or inpatient departments of Wuhan Mental Health Center from September 1,2014 to August 31,2015.At the baseline,the patients and their co-resident family members were interviewed for psychiatric screening and diagnosis,and the family function of each patient's family was assessed by Family Assessment Device (FAD).After clinic service or hospitalization,the patients were followed up by telephone until they recovered from the MDE (within 12 months since the follow-up) or for 12 months if they had not achieved remission.Forty-two mentally healthy subjects,with no family members diagnosis for psychiatric diseases,and matched with MDD patients for age,sex,number of children,family roleand socioeconomic status,were recruited from a community.The family function of the MDD families and the controls were compared by independent sample-T test,and the relationship between family function and duration of the MDE was analyzed by Pearson's correlation.Results:MDD families exhibited higher FAD scores in 5 dimensions than control families except for affective involvement and behavior control (P＜0.01).Patients with relatively good family function showed significantly shorter duration of MDE and higher proportion of remission within 6 months since the follow-up (P＜0.01 and P＜0.05).All the dimensions of FAD demonstrated significant positive correlation with the duration of MDE except for the behavior control.Conclusion:Families with MDD patients show impairments in multiple dimensions of family function,and the family functions of MDD patients are correlated with the prognosis of MDE.Improvement of family function may contribute to better prognosis of MDD.

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BACKGROUND: Hematopoietic stem cells have an ability of multi-directional differentiation and can be differentiated into megakayocytes in the presence of suitable hematopoietic growth factors and lineage special growth factors.Inducing of CD34+ stem cells to megakaryocytes in vitro involves many changes in gene expression, especially the signal transduction genes.OBJECTIVE: To study the expression of signal transduction genes after differentiation from stem cells into megakaryocytes.DESIGN: Open experiment.SETTING: Department of Tumor Immunology, Fujian Provincial Tumor Hospital.MATERIALS: 60-145 mL of cord blood was collected from normal labor fetuses under sterile conditions by using the blood-bag collective method. Patients and their relatives provided the confirmed consent. Reagents and equipments were detailed as follows: VarioMACS segregation apparatus, CD34+ Isolation Kit, SCGM serum-free medium, CD monoclonal antibodies, cytokines, human genome Oligo Set Version 2.1 from the CapitalBio Corporation, and LuxScan 10K/A bi-passage laser scanning apparatus. The researchers obtained consent from the local ethic committee.METHODS: This experiment was carried out in the Department of Tumor Immuonlogy, Fujian Provincial Tumor Hospital and the CaptialBio Corporation. from July 2005 to June 2006. RNA was extracted separately from CD34+ cells purified from cord blood and CD41+ cells induced from the same cord blood. The cDNA probes were synthesized by reverse transcription and purified. CD34+ cells before culturing and CD41+ cells after culturing were labeled with Cy3-dCTP and Cy5-dCTP respectively. Labeled DNA was dissolved in 30 μL hybridization fluid and hybridized at 42 ℃ over night MAIN OUTCOME MEASURES: The differential gene expression between haematopoietic stem cells and megakaryocytes involving cell signal transduction were detected by using DNA microarray analysis. According to the gene chip results,17 differentially expressed genes were selected for further analysis by RT-PCR.RESULTS: In this experiment, 3 522 differentially expressed genes were screened, including 1705 up-regulated genes and 1 817 down-regulated genes. In the 3 522 differentially expressed genes, there were 343 genes involved in cell signaling, 150 genes involved in transcription regulation, 21 genes involved in cell differentiation. The expression of the CD61 gene increased 369.83 fold, the expression of the CD41 gene increased 27.38 fold, and the expression of PF4 gene increased 24.06 folds; The expression of mitogen-activated protein kinases (MAPKs), G protein-coupled receptors (GPCRs) and members of RAS oncogene family increased mostly. The expression of the genes involved in STAT pathways, both SOCS1 and JAK2 were up-regulated, but STAT5A was down-regulated. As determined by the gene chip results, 17 differentially expressed genes were selected for further analysis by RT-PCR. GAPDH was used as the internal control, and RT-PCR results were in agreement and confirmed the finding from the gene chip expression results.CONCLUSION: Thrombopoeitin (TPO) and other hematopoietic growth factors may enhance the proliferation and differentiation of megakaryocytes derived from cord blood stem cells by GPCRs-Ras-MAPK pathway.

OBJECTIVE:To prepare the Albendazole nanoliposomes freeze-dried power and study its properties. METHODS:Freeze-drying method was conducted to prepare Albendazole nanoliposomes freeze-dried power,using the particle size,encapsula-tion efficiency,appearance,redispersibility as indexes,single factor test was combined with orthogonal test to screen freeze-drying preparation technology. The morphological changes,particle size,Zeta potential,moisture content,12 months stability at 4 ℃ be-fore and after freeze-drying were detected. RESULTS:Plus a total content of freeze-dried protective agent was 10%,the ratio of glucose-trehalose-mannitol was 1.0:1.0:3.0,using quick-freeze,pre-freezing 18 h in -35 ℃ refrigerator,dry-freezing 48 h to ob-tain freeze-dried powder. Compared with before freeze-drying,the freeze-dried liposomal morphology had no obvious changes, showing clear phospholipid bilayer membrane structure;the particle sizes before and after freeze-drying were (208.63 ± 1.04) nm and (223.04 ± 2.02) nm,Zeta potentials were (-15.6 ± 0.04) mV and (-19.4 ± 0.06) mV,encapsulation efficiencies were (94.62±0.49)%and(91.10±0.46)%(n=3),respectively. Compared with liposomes,liposomes freeze-dried power had good sta-bility in 12 months at 4 ℃. CONCLUSIONS:Albendazole nanoliposomes freeze-dried power is prepared successfully,its stability is superior to albendazole nanoliposomes,and the freeze-drying technology is feasible.

Objective To analyze the influence of peptidimer-c on the gene expression profiling of K562 cells and investigate the mechanism of peptidimer-c inducing the apoptosis and inhibiting proliferation of K562cells.Methods Trypan blue staining technique was performed for counting the number of living K562 cells treated with peptidimer-c.The ultrastructure changes of K562 cells treated with peptidimer-c was observed under transmission electron microscope.The Human U133 Plus 3.0 gene chips were used to detect the differentially expressed genes of K562 cells treated with peptidimer-c.Reverse transcription PCR was conducted to confirm some genes identified by gene chips.Results Peptidimer-c could induce the apoptosis and inhibit the proliferation of K562 cells.Peptidimer-c caused widely changes of the gene expression profiles of K562 cells.The chip data suggested that there were 529 differentially expressed genes,of which 455 genes were up-regulated and 74 genes were down-regulated.The relevant apoptotic genes were down-regulated markedly,including JUN,AXUD1,TNFRSF10B,etc.Fifteen of the differentially expressed genes were detected by RT-PCR,which was consistent with the chip data.Conclusion Peptidimer-c may induce aooptosis of K562 cells by activating the TNF/TNFR family and the JUN family.

Objective To construct the tranfected cell expressing the human CXCR4 gene and to identify the effect on its immigration. Methods Total RNA was isolated from peripheral blood monouclear cell (PBMC),the full-length CXCR4 gene was amplified by RT-PCR and was inserted into plasmids PBudCE4.1 which have two promtors, after the identification by digestion and sequencing,the recombinant was transfected into K562 cell by lipofectamineTM2000. After screening culture by zeocin, stable transfected K562 cell line was established, and transcription and exression of CXCR4 were checked by flow cytometry; the chemotactic activity of K562 cell transfected and untrandfected CXCR4 was analyzed by Transell plate. Results The eukaryotic expression plasmid PBudCE4.1/ CXCR4 was constructed successfully. The stable trasfected K562/CXCR4 cell lines which highly express CXCR4 was established,the chemotactic activity of K562/CXCR4 was increased significiantly than K562. ConclusionCXCR4 transfected K562 cell line was successfully established, and it can make the basis for the further research on mechanism of extramedullary infiltration in leukemia.

Objective To study the quantitative expression and the correlation of the NF-κB p65,COX-2 and MMP-9 Drotein in the hypopharyngeal carcinoma tissue.Methods FCM method was performed to detect the quantitative expression of the NF-κB p65,COX-2 and MMP-9 protein in 48 cases of hypopharyngeal carcinoma fresh sample and 48 cases of para-carcinoma tissue.Fluorescence Index wasdeftned as the quantitative expression index of the three proteins.Results The quantitative expression of the NF-κB p65,COX-2 and MMP-9 in hypopharyngeal carcinoma tissues(1.16,1.32 and 1.26) was remarkably higher than in para-carcinoma(1.03,1.04 and 1.04).The quantitative expression of three proteins in metastasis group was obviously higher than in non-metastasis group.The expression of NF-κB p65 and COX-2 protein in hypopharvngeal carcinoma tissues was positively related (P＜0.05). Conclusion The high expression of NF-κB p65 and COX-2 is closely related in hypopharyngeal carcinoma tissues.NF-κB p65 might improve the expression of COX-2.

Objective To test experimentally Fluoro-Jade B(FJB) stain method for detecting degeneration of neurons in the basal ganglia. Methods Kainic acid(KA)-lesion model by stereotaxical injection of KA into striatum of rats,MPTP-lesion model by injection of MPTP into intraperitoneal cavity of mice,as well as KA-lesion model of cultured striatal cells were firstly prepared.FJB stain dye was then used to visualize degeneration of neurons in above KA-or MPTP-lesion models. Results KA-or MPTP-induced degenerative neurons including cell bodies and processes could be clearly visualized by FJB stain dye.In the brain sections,FJB-positive stained degenerative neurons were numerously observed in the striatum of KA-lesion rats and the substantia nigra pars compacta of MPTP-treated mice,but not detected in the control animals.Moreover,degenerative neurons were also detected with FJB stain in cultured striatal neurons.Semi-quantitative analysis on percentage(?s) of FJB-positive neurons constituting total cultured striatal neurons in unit area showed that degenerative neurons of KA-lesion group (8.42?1.09)% was evidently more than that of controls (3.42?0.45)%,P

Objective To detect the location of histamine in peripheral sympathetic nerves of guinea pig. Methods Histidine decarboxylase mRNA was detected using in situ hybridization histochemistry with specific oligonucleotide probe,while histamine and tyrosine hydroxylase were detected using double labeled immunohistochemistry with anti-histamine antibody and anti-tyrosine hydroxylase antibody in the superior cervical ganglia of guinea pig. Results The histidine decarboxylase mRNA hybridization signal were detected in both of large and small cells.The TH immunoreactive substance distributed in cytoplasm steadly,but lacked in the nuclei,while the histamine immunoreactive substance distributed in cytoplasm nearby the plasmalemma.After chemical destroy of the guinea pig SCG′s neuron with 6-OHDA,the immunoreactive materials were hardly detected.Conclusion Because the histidine decarboxylase is the only enzyme which catalyzes histidine into histamine,histamine may be synthesized and coexisted with monoaminergic neurotransmitters in the superior cervical ganglia of guinea pig.