Objective To establish a method of large-scale cryopreservation of porcine hepatocytes to meet the need of the biological artificial liver. Methods Porcine hepatocytes were isolated from Chinese miniature swines by collagenase disgestion. The cells were suspended in self-made nutrient solution supplemented with 10% DMSO. The hepatocytes (1?10 10~2?10 10) were stored in liquid nitrogen (-196℃) after being treated with two kinds of freezing methods. The cryopreserved hepatocytes were thawed in 39℃ water after one month and cultured. The morphology, viability and the function of thawed hepatocytes were observed. Results The cell viability of cryobreserved hepatocytes after both freezing methods were high. However, synthesis of urea nitrogen and glucose was higher in stepwise temperature lowering group compared with programmed temperature lowering group. Conclusion The results indicate that the cryopreservation method is feasible and simple, large amount porcine hepatocytes can be cryopreserved.

Objective To clone and express the hexon protein of three prevalent human adenovi -rus strains causing respiratory disease and analyze the antigenic characteristics of the recombinant proteins . Methods The full length genes encoding hexon protein of human adenovirus serotype 3(HAdV3), serotype 4(HAdV4) and serotype 7(HAdV7) were cloned by PCR and sequenced , respectively.The alignment anal-ysis was performed by using hexon gene sequences from GenBank .The major antigenic regions of hexon pro-tein of the three serotypes were expressed in E.coli and purified.The antigenicity, immunogenicity and cross reactivity of the recombinant proteins were determined by ELISA and Western blot assay .Results The full length gene sequence encoding hexon protein of human adenovirus serotype 4 was firstly reported in China , which showed more than 99%homology in both nucleotide and amino acid sequences with the human adeno-virus type 4 NHRC3 strain.The partial hexon protein sequence of HAdV 3, HAdV4 and HAdV7 containing all of the 7 hyperviriable regions ( HVRs) were expressed in E.coli, respectively .The purified recombinant proteins could be recognized by antiserum of the three serotypes of adenovirus .The antiserum samples against the three recombinant proteins could cross-react with particles of the three serotypes of adenovirus . The possible type-and species-specific epitopes were predicted .Conclusion The major antigenic regions of hexon protein of the three serotypes were successfully expressed .The purified recombinant proteins contai-ning both intertypes and type-specific epitopes showed a strong immunogenicity .

To investigate the effect of Yisui Jiedu Recipe (YSJDR), a compound traditional Chinese herbal medicine, on cytokines and their corresponding just another kinase 2-signal transducers and activators of transcription 5 (JAK2-STAT5) signal transduction pathway in bone marrow hematopoietic cells from patients with myelodysplastic syndrome-refractory anemia (MDS-RA).

Objective:To investigate the characteristics of health-related risky behaviors in adolescents with bipolar disorder.Methods:Fifty adolescents aged 12 -18 years,fulfilled the International Statistical Classification of Diseases and Related Health Problems,Tenth Revision(ICD-10)criteria for bipolar disorder(29 in depressive state, 19 in manic state,and 2 in mixed state)and 100 healthy-matched controls were recruited.Their health-related risky behaviors were assessed using the Questionnaire for Adolescents Health-related Risky Behavior Inventory (AHRBI) and the Questionnaire for Adolescents Health-related Risky Behavior Inventory for Parent (AHRBI-P).Results:The bipolar group had higher AHRBI scores in the total scale and six subscales than controls,including Aggression and Violence (AV),Health-Compromising Behavior (HCB),Rule Breaking (RB),Unprotected Sex (US),Self-injury and Suicide (SS),and Smoking and Drinking(SD)[Total scores,55.5(38,119)vs.46(38,65);P <0.05].Besides, the scores of 26 items of bipolar group were all higher than controls(Ps <0.05).The scores in the total scale and six subscales had no difference between AHRBI and AHRBI-P.According to the partial correlation analysis (de-pressive state =0;manic state =1),the AHRBI and AHRBI-P scores of Self-injury and Suicide subscale were nega-tively associated with the episode state (r =-0.32,-0.33;Ps <0.05).The AHRBI scores of'destroy properties'which belongs to the Aggression and Violence subscale were positively associated with the episode state (r =0.32, P <0.05).Conclusion:The adolescents with bipolar disorder have more health-related risky behaviors than the healthy adolescents.The depressive patients have higher risk of suicide.While,the risky behavior of destroying properties tend to occur among the manic patients.

The present study aims to predict the action targets of antidepressant active ingredients of Xiaoyaosan to understand the "multi-components, multi-targets and multi-pathways" mechanism. Using network pharmacology, the reported antidepressant active ingredients in Xiaoyaosan (saikosaponin A, saikosaponin C, saikosaponin D, ferulic acid, Z-ligustilide, atractylenolide I, atractylenolide II, atractylenolide III, paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid and pachymic acid), were used to predict the targets of main active ingredients of Xiaoyaosan according to reversed pharmacophore matching method. The prediction was made via screening of the antidepressive drug targets approved by FDA in the DrugBank database and annotating the information of targets with the aid of MAS 3.0 biological molecular function software. The Cytoscape software was used to construct the Xiaoyaosan ingredients-targets-pathways network. The network analysis indicates that the active ingredients in Xiaoyaosan involve 25 targets in the energy metabolism-immune-signal transmutation relevant biological processes. The antidepressant effect of Xiaoyaosan reflects the features of traditional Chinese medicine in multi-components, multi-targets and multi-pathways. This research provides a scientific basis for elucidation of the antidepressant pharmacological mechanism of Xiaoyaosan.
Antidepressive Agents ; pharmacology ; Benzoates ; Bridged-Ring Compounds ; Coumaric Acids ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; Flavanones ; Glucosides ; Glycyrrhizic Acid ; Lactones ; Medicine, Chinese Traditional ; Monoterpenes ; Sesquiterpenes ; Software

AIM and METHODS: To study the culture met ho d of human neural stem/progenitor cells, these cells derived from human fetal fo rebrains were maintained and expanded in serum-free defined medium containing ba sic fibroblast growth factors (bFGF), epidermal growth factor (EGF) and B27. Whe n they formed neurosphere, these three factors and supplemented FBS were removed to induce differentiation. Cell were cultured for 12-14 d, then fixed for immun ocytochemistry examination. RESULTS: This period of expansion resulted in a 107-fold incre ase in this heterogeneous population of cells. Upon differentiation, they form n eurons, astrocytes and oligodendrocytes, the three main phenotypes in the CNS. CONCLUSION: These results demonstrate the feasibility of long-t erm in vitro expansion of human neural progenitor cells. The advantages of s uch a population of neural precursors for allogeneic transplantation, including t he ability to provide an expandable, well-characterized, defined cell source, ca n form specific neuronal or glial subtypes.

Objective To explore how medical staff in the oncology department understand management of cancer related fatigue (CRF), and explore what factors influencing the effective practice of CRF management. Methods Qualitative inquiry was adopted. Ten medical staffs in the oncology department were selected for in- depth interview. Generic analysis was applied to code, categorize and interpret the qualitative data. Results Participants believed that many factors influenced the CRF, which hadn′t been assessed as an independent symptom and lacked the systematic, effective and specific interventions. The medical staff, patients and their families neglecting the CRF management was the main barrier. Strengthening system construction and staff training was mentioned as major area which needed to be improved. Conclusions CRF management guideline should be formulated according to our national situations based on the clinical practices, besides, the training of correlated clinical knowledge and skills of medical staff should be strengthened and eventually promote the cancer patients′quality of life.

Objective To study the effective surgical methods for the sequela of deep vein thrombosis(DVT) of lower extremity . Methods The clinical data of 32 patients with the sequela of DVT of lower extremity in 36 limbs treated by surgery since 1992 were analysed retrospectively. Results Diagnosis accuracy rate of angiography in sequela of DVT was 100%. There were type I in 5 limbs, type IIA in 4, type IIB in 12,and type III in 15. External constriction valvuloplasty of superficial fomoral vein(SFV) was performed on 16 limbs,valvuloplaasly of SFV on 3 limbs,great saphenous vein popliteal vein anastomosis in situ in 6 limbs,muscular loop plasty on popliteal vein in 4 limbs,common iliac vein SFV anastomosis with artificial vessel interposition in 3 limbs,great saphenoas vien ansatomosed to opposite SFV in 4 limbs. After treatment,symptoms totally disappeared in 28 limbs ( 77.8% ), and improved in 8 limbs. Conclusions The vasography is diagnostic chrysindicatrix of the DVT sequela. Different operations based on the damage of valves should be chosen.

Objective To identify seven species of Gynostemma BI.,including G.pentaphyllum,G. pentagynum,G.cardiospermum,G.longipes,G.yixingense,G.laxiflorum,and G.guangxiense,by in- ter-simple sequence repeat(ISSR)markers.Methods General DNA was isolated from leaves of the seven species in Gynostemma B1.by CTAB,57 primers constituted by ISSR were tested for PCR and sepharose electrophoresis.Results Fourteen primers amplified polymorphic bands,the amplification patterns of primers UBC-873 and UBC-895 were higher in terms of polymorphic and amplified band ratio.They are used to distinguish all the examined seven species.Conclusion ISSR-PCR Method provides a quick,reli- able molecular marker technique for the identification of different species of Gynostemma B1.

OBJECTIVETo study the mechanism underlying the inhibitory effect of Qingluo Tongbi Granule (, QTG) on osteoclast differentiation in rheumatoid arthritis in rats.

METHODSFibroblast and monocyte co-culture were used to induce osteoclast differentiation in adjuvant-induced arthritic (AIA) rats. Serum containing QTG was prepared and added to the osteoclasts, and activation of the tumor necrosis factor receptor-associated factor 6/mitogen-activated protein kinase/nuclear factor of activated T cells, cytoplasmic1 (TRAF6/MAPK/NFATc1) pathways was examined.

RESULTSThe induced osteoclasts were multinucleated and stained positive for tartrate-resistant acid phosphatase (TRAP) staining. Serum containing QTG at 14.4, 7.2 or 3.6 g/kg inhibited the activation of TRAF6, extracellular regulated protein kinase (ERK)1/2, c-Jun N-terminal kinase (JNK) and p38 and decreased the percentage of cells with nuclear NFATc1 in a dose-dependent manner, the high and middle doses exhibited clear inhibitory activity (P<0.01 and P<0.05, respectively). After the addition of MAPK inhibitors, the NFATc1 expression showed no significant difference compared with the control group (P>0.05).

CONCLUSIONSSerum containing QTG could generally inhibit the TRAF6/MAPK pathways and possibly inhibit the NFATc1 pathway. In addition, QTG may regulate other signaling pathways that are related to osteoclast differentiation and maturation.

Adjuvants, Immunologic ; adverse effects ; Animals ; Arthritis, Experimental ; pathology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Coculture Techniques ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; pathology ; Male ; Monocytes ; pathology ; Osteoclasts ; cytology ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane ; pathology