Objective To develop a selective isolation media which has a high isolation ratio for Legionella.Methods We compared the growth of Legionella pneumonia on BCYE? plate (with DGVP), BCYE?(with GVPC) and BCYE? (with CCCV) by colony count and compared with BCYE? plate. The comparison of isolation ratio of Legionella pneumonia from air-conditions cooling water with 3 kinds of BCYE? plate containing different anti-reagents was performed.Results The results of colony count on BCYE? plate (with DGVP) and BCYE?(with GVPC) was identical with that on BCYE? plate at same concentration. But the result of colony count on BCYE? plate (with CCCV) was half of the former 2 kinds of plate. 5 strains of Legionella pneumonia isolated from 9 central air-condition cooling water were used by BCYE? plate (with DGVP) and BCYE? (with GVPC), while only 2 strains of Legionella pneumonia were isolated by BCYE? plate (with CCCV). Legionella pneumonia wasn′t detected by BCYE? plate.Conclusion BCYE? plate with antibiotics system of CCCV showed obvious suppression to Legionella pneumonia, and so that there was a low isolation ratio to Legionella pneumonia. But BCYE? plate with antibiotics system of DGVP and GVPC had a few suppression and showed a high isolatio ratio for Legionella pneumonia. Moreover, BCYE? plate with antibiotics system of DGVP and GVPC can effectively inhibit non-Legionella and increase isolation ratio of Legionella. They should be the first selection media for Legionella from environment and clinical samples.

The effects of phorbol-12,13-dibuterate (PDBu) on total sodium current (I(Na)-total), tetrodotoxin-resistant sodium current (I(Na)-TTXr), 4-AP-sensitive potassium current (I(A)) and TEA-sensitive potassium current (I(K)) in trigeminal ganglion (TG) neurons were investigated. Whole-cell patch clamp techniques were used to record ion currents in cultured TG neurons of rats. Results revealed that 0.5 micromol/L PDBu reduced the amplitude of I(Na)-total by (38.3+/-4.5)% (n=6, P<0.05), but neither the G-V curve (control: V (0.5)=-17.1+/-4.3 mV, k=7.4+/-1.3; PDBu: V (0.5)=-15.9+/-5.9 mV, k=5.9+/-1.4; n=6, P>0.05) nor the inactivation rate constant (control: 3.6+/-0.9 ms; PDBu: 3.6+/-0.8 ms; n=6, P>0.05) was altered. 0.5 micromol/L PDBu could significantly increase the amplitude of I(Na)-TTXr by (37.2+/-3.2)% (n=9, P<0.05) without affecting the G-V curve (control: V (0.5)=-14.7+/-6.0 mV, k=6.9+/- 1.4; PDBu: V (0.5)=-11.1+/-5.3 mV, k=8.1+/-1.5; n=5, P>0.05) or the inactivation rate constant (control: 4.6+/-0.6 ms; PDBu: 4.2+/-0.5 ms; n=5, P>0.05). 0.5 mumol/L PDBu inhibited I(K) by (15.6+/-5.0) % (n=16, P<0.05), and V (0.5) was significantly altered from - 4.7+/-1.4 mV to -7.9 +/-1.8 mV (n=16, P<0.05). I(A) was not significantly affected by PDBu, 0.5 mumol/L PDBu decreased I(A) by only (0.3+/-3.2)% (n=5, P>0.05). It was concluded that PDBu inhibited I(Na)-total but enhanced I(Na)-TTXr, and inhibited I(K) without affecting I(A). These data suggested that the activation of PKC pathway could exert the actions.

Objective To investigate the effects of the polymorphisms in the promoter of ATP binding cassette transporter (ABCG1) on the transcription activity,and the relationship of the polymorphisms with the susceptibility to coronary artery disease (CAD).Methods A case-control study was conducted,217 CADpatients and 142 controls were enrolled in this study.Thesingle nucleotide polymorphisms (SNPs) in the promoter of ABCG1 were identified by sequencing.The promoter haplotypes of ABCG1 were determined with allele specific primer sequencing or Gene cloning sequencing.The transcription activity of the promoter haplotypes were evaluated with dual luciferase reporter system.The frequency of SNPs and haplotypes were analyzed between CAD group and the control group,premature CAD and non-premature CAD group,as well as multivessel lesion and single vessel lesion group.The frequency distribution was compared between two groups with x2 test or Fisher exact test.The difference of the luciferase activity was compared between groups by t-test or one-way analysis of variance.Results Only 3 SNPs were found in ABCG1 promoter sequence of about 1 000 bp upstream of the transcription start site,which are-384 (A/G),-204 (A/C) and-134 (T/G),respectively.The 3 SNPs are in strong linkage disequilibrium,Tajima's D =2.655 (P ＜ 0.01),which constituted 3 haplotypes.There was no significant difference in SNPs and haplotype frequency between the CAD group and the normal control group,and the severity of vascular disease and the early onset of coronary heart disease were not associated with the polymorphisms in ABCG1 promoter.There was no significant difference in the transcriptional activity of the three constitutive promoter haplotypes,but the transcriptional activity was notably elevated as the GAT haplotype was mutated into GAG (P ＜ 0.05).Conclusions The 3 SNPs identified in ABCG1 promoter region A did not alter the promoter activity.There was no significant correlation between the frequency distribution of SNPs and promoter haplotypes and the susceptibility to CAD.

Objective To investigate the value of cytokinesis-block micronuclei(CBMN)assay in estimation of the biological doses of the victims of radiation accident.Methods Samples of peripheral blood were collected from the 5 victims(Subjects 1-5)at 16 h after the radiation accident of Taiyuan,Shanxi Province.And the peripheral blood samples and bone marrow sample were collected from the victim No.1 at 23 and 24 h after the radiation.Eight days after the accident Subject 1 underwent allogeneic peripheral blood hematopoietic stem cell transplantation.At difierent time points in the period of 1 year after the accident.peripheral blood samples were collected from these 5 victims.CBMN assay was conducted on the peripheral blood lymphocytes on the samples,and the biological doses were estimated based on the micronuclei(MN)frequencies.The nuclear division index(NDI)obtained from in vitro irradiation experiment using high dose of 60Coγ-rays was used to estimate the exposed doses for Subject 1. Dynamic arialysis of the MN frequency for the 5 victims was performed in the period of 1 year after the accident.Results The MN frequency of Subject 1 surpassed the value corresponding to the upper limit of the MN dose.effective curve.The dose range estimated bv the combination of the CBMN and NDI (CBMN+NDl)assay was 10-20 Gy for Subject 1.The doses estimated by MN frequency for Subjects 2,3,4,and 5 were 3.6,2.9,2.3,and 2.9 Gy,respectively.The estimated doses were in accordance with those estimated by physicat method.chromosome aberration analysis.and clinical symptoms.Prominent decrease of the MN frequency was observed at 26 d after the accident(18 d after the transplantation)for Subject 1(u=3.295,P<0.05).Gradual decrease of MN frequency was observed after the accident for Subjects 2,3,4,and 5.The MN frequencies 1 month after the accident of Subjects 3,4,and 5 were all significantly lower than those 16 h later(u=6.874,4.526,and 7.811,P<0.05).Conclusions Quick and accurate.CBMN assay reinforces and verifies the result of chromosome aberration analysis.The new index CBMN+NDI assay is of reference valne for estimating higher dose of irradiation.

To further investigate the mechanisms of action of icariin (ICA), we assessed the effects of ICA on the in vitro formation of cGMP and cAMP in isolated rabbit corpus cavernosum. Isolated segments of rabbit corpus cavernosum were exposed to increasing concentrations of ICA and the dose-dependent accumulation of cGMP and cAMP was determined in the tissues samples by means of 125I radioimmunoassay. Responses of the isolated tissues preparations to ICA were compared with those obtained with the reference compounds sildenafil (Sild). Furthermore, the effects of ICA on the mRNA expression of specific cGMP-binding phosphodiesterase type V (PDE5) in rat penis were also observed. After incubation with ICA for 6 h or 14 h respectively, the levels of PDE5 mRNA were examined by reverse transcriptase polymerase chain reaction (RT-PCR). The results showed that ICA increased cGMP concentrations directly (P < 0.05), but there was no significant effect on cAMP concentrations (P > 0.05). In the presence of sodium nitroprusside (SNP), a stimulatory agent of cGMP, both ICA and Sild increased cGMP concentrations with increasing dose (P < 0.01). Their EC50 was 4.62 (ICA) and 0.42 (Sild) micromol/L respectively. Under the same condition, ICA and Sild unaltered cAMP level significantly (P > 0.05). There were PDE5A1 and PDE5A2 mRNA expressions in rat corpus cavernosum with PDE5A2 being the dominant isoform. ICA could obviously inhibit these two isoforms mRNA expression in rat penis, and decrease PDE5A1 more pronouncedly (P < 0.01). The present study indicated that the aphrodisiac mechanisms of icariin involved the NO-cGMP signal transduction pathway, with increasing cGMP levels in the corpus cavernosum smooth muscle. The inhibitory effect of icariin on PDE5 mRNA expression, especially on PDE5A1, might account for its molecular mechanisms for its long-term activity.

The effects of phorbol-12,13-dibuterate (PDBu) on total sodium current (INa-total), tetrodotoxin-resistant sodium current (INa-TTXr), 4-AP-sensitive potassium current (IA) and TEA-sensitive potassium current (IK) in trigeminal ganglion (TG) neurons were investigated.Whole-cell patch clamp techniques were used to record ion currents in cultured TG neurons of rats. Results revealed that 0.5 μmol/L PDBu reduced the amplitude of INa-total by (38.3±4.5)% (n=6, P＜0.05), but neither the G-V curve (control: V0.5 =-17.1±4.3 mV, k=7.4±1.3; PDBu: V0.5=-15.9±5.9 mV, k=5.9±1.4; n=6, P＞0.05) nor the inactivation rate constant (control: 3.6±0.9 ms; PDBu: 3.6±0.8 ms; n=6, P＞0.05) was altered. 0.5 μmol/L PDBu could significantly increase the amplitude of INa-TTXr by (37.2± 3.2)% (n=9, P＜0.05) without affecting the G-V curve (control: V0.5=-14.7±6.0 mV, k=6.9±1.4; PDBu: V0.5=-11.1±±5.3 mV, k=8.1±1.5; n=5, P＞0.05) or the inactivation rate constant (control: 4.6±±0.6 ms; PDBu: 4.2±0.5 ms; n=5, P＞0.05). 0.5 μmol/L PDBu inhibited IK by (15.6±5.0) % (n=16, P＜0.05), and V0.5 was significantly altered from - 4.7±1.4 mV to -7.9 ±1.8 mV (n=16, P＜0.05). IA was not significantly affected by PDBu, 0.5 μmol/L PDBu decreased IA by only (0.3±3.2)% (n=5, P＞0.05). It was concluded that PDBu inhibited INa-total but enhanced INa-TTXr, and inhibited IK without affecting IA. These data suggested that the activation of PKC pathway could exert the actions.

After healing of burn wound, skin of scar, transplanted skin grafts, and healed donor site wound suffer from temporary or permanent loss of function of sebaceous glands and dysfunction of skin surface lipid film formation, resulting in desiccation, desquamation, and sensitiveness of the skin, making areas of newly formed skin unsatisfactory. Therefore a good rehabilitation may fail. In this paper, the composition, physiochemical properties, and reconstruction of skin surface lipid film are discussed.
Cicatrix ; Lipids ; physiology ; Skin ; pathology ; Skin Transplantation ; Wound Healing

To investigate the effects of WIN 55,212-2 on IK in cultured rat trigeminal ganglion (TG) neurons, whole-cell patch clamp techniques were used to record the IK before and after WIN 55,212-2 perfusion at different concentrations. 30 μmol/L WIN 55,212-2 markedly (35.7 %±7.3%, P＜0. 01, n=8) inhibited IK currents, and the currents were partially recovered after washing.30μmol/L WIN 55,212-2 also induced a significant depolarizing shift in conductance-voltage parameters (control: V0.5 =10.43 ± 4.25 mV, k= 16.27±3.86; WIN 55,212-2: V0. 5 =24.71±3.91mV, k =16.69±2.75; n = 8, P＜0.01 for V0. 5). 0.01μmol/L WIN 55,212-2 slightly (27.0 %± 7.9 %, P＜0.05, n=7) increased IK currents, but had no significant change in conductance voltage parameters (control: V0.5 =10. 74±5. 27 mV, k=17. 33±2. 96; WIN 55,212-2: V0.5 =11.06±2.05 mV, k=19. 69±6. 60; n=7, P＞0.05 for V0.5 and k). These results suggested that WIN 55,212-2 has dual action, which might be through different receptors.

To further investigate the mechanisms of action of icariin (ICA), we assessed the effects of ICA on the in vitro formation of cGMP and cAMP in isolated rabbit corpus cavernosum. Isolated segments of rabbit corpus cavernosum were exposed to increasing concentrations of ICA and the dose-dependent accumulation of cGMP and cAMP was determined in the tissues samples by means of 125I radioimmunoassay. Responses of the isolated tissues preparations to ICA were compared with those obtained with the reference compounds sildenafil (Sild). Furthermore, the effects of ICA on the mRNA expression of specific cGMP-binding phosphodiesterase type V (PDE5) in rat penis were also observed. After incubation with ICA for 6 h or 14 h respectively, the levels of PDE5 mRNA were examined by reverse transcriptase polymerase chain reaction (RT-PCR). The results showed that ICA increased cGMP concentrations directly (P＜0.05), but there was no significant effect on cAMP concentrations (P＞0.05). In the presence of sodium nitroprusside (SNP), a stimulatory agent of cGMP,both ICA and Sild increased cGMP concentrations with increasing dose (P＜0.01). Their EC50 was 4.62 (ICA) and 0.42 (Sild) μmol/L respectively. Under the same condition, ICA and Sild unaltered cAMP level significantly (P＞0.05). There were PDE5A1 and PDE5A2 mRNA expressions in rat corpus cavernosum with PDE5A2 being the dominant isoform. ICA could obviously inhibit these two isoforms mRNA expression in rat penis, and decrease PDE5A1 more pronouncedly (P＜ 0.01). The present study indicated that the aphrodisiac mechanisms of icariin involved the NO-cGMP signal transduction pathway, with increasing cGMP levels in the corpus cavernosum smooth muscle. The inhibitory effect of icariin on PDE5 mRNA expression, especially on PDE5A1, might account for its molecular mechanisms for its long-term activity.