Multidetector computed tomography (CT) has transformed CT from an transaxial cross-sectional technique into a genuine 3D imaging that allows for arbitrary cut planes as well as excellent 3D displays of the volume data. Multidetector CT scanners contribute to reduced scanner time, decreased section collimation and increased scan length.

Objective To obtain the endophytic fungi which produce Huperzine A from four species in Hupziaceae and improve the culture conditions of screening endophytic fungi. Methods Plant materials were cultivated in culture medium after sterilization and their endophytic fungi were isolated. Hupzine A from metabolic was determined by HPLC, and the strains were identified by microscopic features. Results Thirty-two endophytic fungi were isolated from Huperzia serrata (Thunb. ex Murray) Trev, H. serrata (Thunb. ex Murray) Trev var. longipetiolata (Spring) H. M. Chang, H. appressa (Desv.) Love et D. Love, Phlegmariurus cryptomerianus (Maxim.) Ching ex L. B. Zhang et H. S. Kung. Two of these strains were isolated from P. cryptomerianus, HA15 (Blastomyces sp.) and HA23 (Botrytis sp.), which produced Huperzine A. Conclusions Endophytic fungi producing Hupzine A has been successfully isolated from P. cryptomerianus.

Objective To obtain recombinant human granulysin using prokaryotic expression system.MethodsTotal RNA was extracted from cultured PBMC. Granulysin gene segments were obtained with granulysin-specific primers by RT-PCR and then inserted into pET32a(+) plasmid. After identification by DNA sequence,pET-GNLY9K and pET-GNLY15K were transferred to E. coli Rosetta (DE3). The fusion protein was identified by SDS-PAGE and Western blot.The bioactivity of granulysin fusion protein was measured by MTT assay.Results The prokaryotic expression vectors pET-GNLY9K and pET-GNLY15K were successfully constructed.The corresponding protein was highly expressed in E.coli. Recombinant protein was specifically bound by anti-granulysin antibody. GNLY9K fusion protein significantly inhibited the growth of A549 cells in a dose-dependent manner,while GNLY15K had little effect on the growth of A549.Conclusion Granulysins with different mw were successfully expressed using prokaryotic expression system,which might be helpful for the further study of granulysin.

Objective To investigate the effects of lactoferrin on activity of PKG in spinal dorsal horn in a rat model of neuropathic pain(NP).Methods Thirty-two male SD rats weighing 200-250 g were randomly divided into4 groups(n = 8 each): sham operation group(group S),NP group,lactoferrin group and KT5823(an inhibitor of PKG)group.Neuropathic pain was produced by placing loosely constrictive ligatures around the common sciatic nerve in group NP,lactoferrin and KT5823,while the sciatic nerve was only exposed but not ligated in groupS.In group S and NP,normal saline 10 μl + 50% dimethyl sulfoxide(DMSO)10 μl were injected intrathecally.Lactoferrin 100 μg + 50% DMSO 10 μl were given intrathecally in group lactoferrin.Lactoferrin 100 μg + KT5823 10 μl were given intrathecally in group KT5823.The paw withdrawal latency(PWL)to a thermal nociceptive stimulus was measured every 30 min within 180 min after administration.The rats were then sacrificed and the spinal cord was removed.The activity of PKG in the spinal dorsal horn was determined by immunofluorescence.Results Compared with group NP and KT5823,the PWL was significantly prolonged after administration in group lactoferrin and the PKG activity was significantly increased in group lactoferrin(P ＜ 0.05).There was no significant difference in the parameters mentioned above between group NP and group KT5823(P ＞ 0.05).Conclusion Lactoferrin reduces NP by inhibiting the activity of PKG in spinal dorsal horn in rats.

Adult ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Peripheral Vascular Diseases ; drug therapy ; Phytotherapy ; Takayasu Arteritis ; drug therapy

Objective To investigate protective effects and mechanism of folic acid on brain neural cells in preeclampsia rat model.Methods Adult pregnant Wistar rats were randonly divided into 4 groups (n = 10 in each group).Rats in model group were injected intraperitoneally with homocysteine (Hcy,200 mg · kg-1 · d-1) daily and were injected subcutaneously every other day with monosodium glutamate (MSG,1 g · kg-1 · 48 h-1) from the 10th day of pregnancy to establish the model of preeclampsia. Lowdose folic acid (low dose group 10 ng · kg-1· d-1) and high-dose folic acid (high dose group 20 mg · kg-1 · d-1 ) were given intragastric administration with folic acid tablets dissolved in saline daily at the same time of establishing model.Rats in control group were injected or intragastric administration with the same dose of saline as above up to the 20th day of pregnancy.Brain tissue was fixed on the 20th day of pregnancy, so was that plasma folic acid was measured with automatic electro-chemiluminescence.Rats' immunohistochemical staining.bcl-2 mRNA and protein expression changes were observed by using reverse transcription(RT) -PCR and western blot.Results ( 1 ) Plasma folate concentrations were ( 39.5 ± 3.4 )nmol/L in low dose group and (40.1 ±5.4) nmol/L in high dose group, which were all significantly higher than (26.9 ± 6.7 ) nmol/L in model group( P ＜ 0.01 ).Plasma folate in low dose and high dose group did not show significant difference( P ＞ 0.05 ); ( 2 ) Apoptosis cell were 48.2 ± 9.1 in low dose group and 44.7 ±8.3 in high dose group, which were significantly lower than 75.8 ± 10.1 in model group (P＜0.01).However, apoptosis cell in low dose and high dose group did not show significant difference( P ＞0.05 ) ;(3 )significant difference( P ＞ 0.05 ); (4) bcl-2 mRNA and protein expression were 0.98 ± 0.49 and 0.89 ±0.52 in low dose group and 0.95 ± 0.38 and 0.92 ± 0.47 in high dose group which was significantly higher than 0.62 ± 0.20 and 0.45 ± 0.37 in model group ( P ＜ 0.01 ); bcl-2 expression in low dose and high dose group showed no significant difference ( P ＞ 0.05 ).Conclusions Folic acid has a protective role on neural activation and promoting bcl-2 gene and protein expression.

Objective To obtain recombinant human granulysin using prokaryotic expression system. Methods Total RNA was extracted from cultured PBMC. Granulysin gene segments were obtained with granulysin-specific primers by RT-PCR and then inserted into pET32a(+) plasmid. After identification by DNA sequence, pET-GN-LY9K and pET-GNLY15K were transferred to E. Coli Rosetta (DE3). The fusion protein was identified by SDS-PAGE and Western blot. The bioactivity of granulysin fusion protein was measured by MTT assay. Results The prokaryotic expression vectors pET-GNLY9K and pET-GNLY15K were successfully constructed. The corresponding protein was highly expressed in E. Coli. Recombinant protein was specifically bound by anti-granulysin antibody. GNLY9K fusion protein significantly inhibited the growth of A549 cells in a dose-dependent manner, while GN-LY15K had little effect on the growth of A549. Conclusion Granulysins with different mw were successfully expressed using prokaryotic expression system, which might be helpful for the further study of granulysin.

Platelet activating factor (PAF) is a biologically active membrane phospholipid mediator. In this study we investigated the effects of PAF on the lymphocyte proliferation, production of IL-2, natural killer cytotoxic factor (NKCF) and colony stimulating factor (CSF) by BALB/c mice splenocytes in vitro. The results showed that PAF inhibited lymphocyte proliferation at concentrations of 10-10~ 10-7 mol/L. CTLL-2, YAC-1 and munne bone marrow cells were used for the assays of IL-2, NKCF and CSF, respectively. Production of IL-2, NKCF and CSF from ConA stimulated murine splenocytes was significantly inhibited by PAF at 10-7 mol/L. Inhibition of spleen cells proliferation and secretion may play an important role in the pathological effects of PAF.

Objective To evaluate the safety and feasibility and clinical effectiveness of living relative donor kidney transplantation(LDKT)and summarize its clinical experience. Methods The clinical data of 30 cases of LDKT were retrospectively analyzed.Except for 2 cases being donated by spouse,the others were donated by blood relative donors.6 cases shared two haplotypes,and 22 cases shared one haplotype,and one case 4 mismatched,and 1 fully mismatched.All donors underwent open nephroectomy,in which 7 cases donated right kidneys and 23 donated left kidneys.In 30 cases of recipients,1 case received cadaver donor kidney transplantation and lost her allograft because of superacute rejection.Triple-combined immunosuppressive protocols consisted of calcineurin inhibitor (CNI),mycophenolate mofetil(MMF)or azathioprine(AZa)and steroid. Results All donors'hospital stay was 7 to 10 days postoperatively without any surgical complications. All donors kept their normal kidney function within 3 to 6 months'follow-up.Except for 1 case of death because of lung in fection,29 cases of recipients survived,in which 28 cases kept their normal function kidney within 1 to 4 years of follow-up and 1 case occurred chronic allograft nephropathy after one year.Except one case of DGF,29 cases of recipients retained their normal kidney function in 3 to 5 days postoperatively.Rejection episodes occurred in 4 cases,of which 3 cases were reversed by methylprednisone and 1 case by antithymocyte globulin(ATG)and Tacrolimus.Pneumonia developed in 3 cases,of which 2 cases were cured and 1 case failed.Hematoma was found around allograft in 1 case and wag surgi cally removed.Urinary leakage was happened in 2 cases of recepients and were cured by conservative treatment. Conclusions LDKT is safe and feasible with good long-term results and more advantages such as optimal HLA matches and less ischemia time and lower acute rejection,low-dose immunosup pressants.

Objective To establish a stable primary culture of rat cerebellar granule neurons in vitro for further study the toxic effects of chronic arsenic exposure on cerebellar cells.Methods Cerebellar cortices were taken from brain of Wistar rat 5-7 day old after born under stereoscopic microscope.Single cell suspension was acquired after digestion and washing with trypsin (0.25％) and DNase Ⅰ solution,respectively.Granule cells were purified from other cells by differential velocity adherence method for two times.Rat cerebellar granule neurons were seeded in culture plate pre-coated with poly-L-lysine.Neurons growth,development and synaptic connections were observed daily.The neurons were identified by neuron specific enolase (NSE) immunofluorescence technique.Results The neurons were affixed to the culture plate in 24 hours,in reticular arrangement observed under contrast microscope.Granule cells gradually turned round from oval and outlines became clearer in 2-3 days.In 4-6 days,there were a wide range of synaptic connections among the neurons and a mature nerve cell network formed.A large quantity of cerebellar granule neurons was seen by NSE identification.Few bigger cells such as purkinjes cells and glial cell outlines were also seen in the same visual field.Conclusions This is a successful primary culture method for acquirement of rat cerebellar granule neurons.The method can provide experimental basis for future studies the toxic effects of chronic arsenic exposure on cerebellar cells.