Objective To investigate the effects of paeoniflorin on proliferation and the protein expression of HFCL of human bone marrow stromal cells and to discuss molecular mechanism of blood enriching functions of paeoniflorin. Methods Flow cytometry and proteomics were used respectively to measure the effect of paeoniflorin on cell cycle and protein expression of HFCL. Results Paeoniflorin could promote HFCL from G_0/G_1 phase to S phase, increase proliferative index, up-regulate nine kinds of proteins, and down-regulate five kinds of proteins of HFCL. The proteins up-regulated include Ras-related nuclear protein, lamin A/C, isocitrate dehydrogenase 3 (NAD+), triosephosphate isomerase, ATP synthase, ribosomal protein P2, and chaperonin containing t-complex polypeptide 1. Conclusion Through promoting HFCL proliferation, acting on multiple protein targets, enhancing the synthesis of cytoarchitecture proteins and the expression of protein chaperonin, increasing the energy metabolism of HFCL, and suppressing the apoptosis of HFCL, paeoniflorin plays indirectly blood enriching function.

Objective Summarize anterior open tension-free inguinal hemiorrhaphy, especially application experience and understanding in the 952 cases of day surgery, and clarify the advantages. Method During December 2004 - June 2007, we treated 952 Patients of inguinal hernia and femoral hernia,using local anesthesia, tension-free, in the form of day surgery. And the resulte were compare to traditional hemiorrhaphy, tension-free hemiorrhaphy of hospital in all aspects. Results Patients in this group were all cured.Intraoperative and postoperative pain was mild.All patients should be used only a small amount of postoperative oral analgesics, and no urinary retention. They got out of bed immediately after surgery, and they recorered fast ,with less complications. They could be discharged home in surgical day (2 h later). Af ter 18 months follow-up, only 2 cases recurrence. Conclusions (1) Compared with the traditional hemiorrhaphy and ambulatory tension-free hemiorrhaphy, tension-free during the day has lots of advantages, such as easier method, wider surgical indications, faster postoperative recovery, shorter hospital stay, less complications and lower recurrence rate;(2)Tension-free hernion'haphy day surgery is safe，feasible，and has obvi-ous advantages.

Objective To optimize the experiment conditions of surface-enhanced Raman spectroscopy detection of serum fingerprint spectra.Methods Normal human serum was used as the sample and Ag nanoparticles as the active substrate.The enhanced signals of different optimized experiments were obtained , including serum dose(2.5 to 500 μl), incubation time(10 to 30 minutes) temperature(4℃,room temperature and 37℃),and different treatment(extraction and protein removal).Results and Conclusion Serum doses should not exceed 50μl.The ratio should range from 1∶1 to 5∶1, the incubation time is from 10 to 30 minutes, and the incubation temperature from 4℃ to 37℃.The signals of samples directly mixed with an active substrate are stronger than those of samples which are extracted or protein removed .

Recently surface-enhanced Raman spectroscopy (SERS) has been widely used in physics, chemistry and bio-medical science.Due to its high sensitivity and specificity,SERS is often used to detect changes in serum components in humans.Various biomolecules in human serum, such as proteins, lipids and nucleic acids, have their own distinctive raman spectroscopy so that different raman shift, band intensity and width reflect different metabolic abnormalities of cells at the molecular level in human serum.In this paper we described the general situation of SERS and summarized the latest research progress in a variety of diseases of human serum.Prospects of developmenls are also outlined.

Objective To develop mycobacterial inducible expression vectors which permit to overexpress Mycobacterium tuberculosis (Mtb) immunodominant antigen, and to analyze its immunogenicity after purification by affinity chromatography. Methods The regulatory region of M. smegmatis (Ms) acet-amidase(pACE) was obtained by PCR amplification, and was used as promoter to construct the mycobacteri-al inducible expression vectors, pMF series. The coding gene of Mtb chimeric antigen Ag856A2 which is a recombinant Ag85A with 2 copies of ESAT-6 inserted in its Acc Ⅰ site and showed excellent immunogenicity in the animal experiments we described previously, was cloned into the pMF vector series, and was induced to express by the addition of acetamide. The recombinant protein expressed in the Ms was purified by the Ni~(2+)-NTA affinity chromatography, the resulted homologous recombinant antigen was added into the spleen cells separated from BCG vaccinated mice, and the immunogenicity was analyzed by the IFN-γ ELISPOT as-say. Results The mycobacterial inducible expression vectors, pMF series was constructed successfully, target antigen could be. induced to express in the Ms by the addition of 0.02% acetamide, and could be puri-fied by the Ni~(2+) -NTA affinity chromatography due to the addition of 6×His tag in the vector pMF406. Fur-thermore, the mycobactefial homologous antigen could induce more IFN-γ secretion than the heterogonous one. Conclusion The mycobacterial inducible expression system based on the regulatory region of Ms acet-amidase as promoter could permit the Mtb target antigen of interest overexpression and purification, and the immunogenicity of the homologous antigen from Ms is better than that of be expressed from E. coli, which may be more potential for immunological detection of tuberculosis.

Objective To study the anti-tumor effects of alcohol extraction of Coix stalk objects on H22 tumor-bearing mice. Methods The animal model of tumor bearing mice with H22 ascitic tumor cells was established. Eighty-four model mice were randomly and equally divided into Coix stalk extract groups 1-5 (10, 8, 6, 4 and 2 g/kg), model control group and cyclophosphamide group. Mice were treated orally with Coix stalk alcohol extraction solution (10, 8, 6, 4 and 2 g/kg), cyclophosphamide 0.02 g/kg and normal saline once a day for 8 days for Coix stalk extract group, cyclophosphamide group and model control group. The mouse activity, the size and the appearance of time of abdominal swelling, and changes of hair, feeding and drinking water quantity were observed in groups of mice. The solid tumor mass was measured in H22 tumor-bearing mice. The tumor inhibitory rate, liver index, spleen index and thymus index were calculated. Results The axillary tumor muster was found first in model control group with the fastest growth, reduced independent activity, decreased appetite and dim in hair color, followed by the Coix stalk extract group 1 and group 2. The last was Coix stalk extract group 5 and cyclophosphamide group. The solid tumor mass were (0.47±0.18), (0.37± 0.13), (0.34±0.10), (0.30±0.11) and (0.28±0.09) mg for Coix stalk alcohol extract groups 1-5, which were significantly lower than those of model control group (0.60 mg±0.21 mg, F=5.700,P<0.05). The tumor inhibition rates were 21.67%, 38.33%, 43.33%, 50.00%, 53.33%and 60.00%in Coix stalk extract groups 1-5 and cyclophosphamide group. The liver index, spleen index and thymus index were lower in cyclophosphamide group and Coix stalk alcohol extract groups than those of model control group (except for the spleen index of Coix stalk extract group 1). The liver index was lower in Coix stalk ethanol extract groups than that of cyclophosphamide group. There were no significant differences in the spleen index, thymus index between Coix stalk ethanol extract groups and cyclophosphamide group. Conclusion Coix stalk alcohol extract has inhibitory effects on the tumor and liver damage in H22 mice.

OBJECTIVETo evaluate the inhibitory effects of HCV antisense oligonucleotide drugs in vivo.

METHODSTransgenic mice were generated by microinjection. The construct of luciferase controlled by HCV 5'NCR that contains CMV promotor was injected into the male pronuclus of fertilized eggs of ICR mice.

RESULTSSixty-eight survival birth transgenic mice were identified by PCR amplification with tail DNA, 13 of whom were positive with an integration ratio of 19.2% (13/68). Transgenic mRNA was detected by RT-PCR in the tissue of three mice's offspring that contain transgenic DNA. Luciferase expression was detected in a line (35#) using the luciferase assay system and the expression persisted in the F2 generation. The phenotype of the mice in this line was normal and there was no significant difference in physiology from normal mice.

CONCLUSIONSThis line of transgenic mice will provide a useful animal model for the study of function of HCV 5'NCR and the evaluation of HCV antisense drugs in vivo.

Animals ; Artificial Gene Fusion ; methods ; Drug Evaluation ; Hepacivirus ; drug effects ; genetics ; Luciferases ; biosynthesis ; genetics ; Mice ; Mice, Inbred ICR ; Mice, Transgenic ; Microinjections ; methods ; Models, Animal ; Promoter Regions, Genetic ; genetics ; RNA, Antisense ; pharmacology ; RNA, Messenger ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Zygote ; cytology ; growth & development