In order to probe the effects of lipopolysaccharide(LPS) on ? adrenoceptor (? AR) and ? AR kinases in rat pulmonary microvascular endothelial cells (RPMVEC), radioactive ligand binding assay was used to measure the distribution density of ? AR before and after LPS acting in it; Western blot was used to observe the expression changes in ? AR kinases, including GRK2 and GRK3. The results showed that the maximum binding capacity (Bmax) of RPMVEC ? AR was 5 583?0 306 fmol/10 5 cells. The Bmax of ? AR in LPS group significantly decreased compared with normal control group ( P

AIM　Observing the influence of lipopolysaccharide (LPS) on changes of the permeability and actin cytoskeleton of rat pulmonary microvascular endothelial cells (PMVEC) monolayer, we explore their relationship with β-adrenoceptor(β-AR) and G protein subunit(Gsα),and the interfering action of anisodamine. METHODS　PMVEC was isolated and cultured from Wistar rat in vitro, radioligand binding assay and flow cytometer were used. RESULTS　After incubation of LPS in vitro, the central F-actin of PMVEC depolymerized and its density decreased obviously ,while the permeability of PMVEC monolyer increased significantly. Meanwhile it was found that LPS can induce the down regulation of β-AR and Gsα protein level. Anisodamine can inhibit these changes above. CONCLUSION　LPS can directly cause the increase of permeability of PMVEC monolayer which relates to the depolymerization of central F-actin in PMVEC. Anisodamine may take part in regulating actin cytoskeleton of PMVEC and attenuate LPS-induced the increase of permeability of PMVEC monolayer by influencing two links of both β-AR and Gsα protein.

AIM Observing the influence of lipopolysaccharide (LPS) on changes of the permeability and actin cytoskeleton of rat pulmonary microvascular endothelial cells (PMVEC) monolayer, we explore their relationship with ?-adrenoceptor(?ＡＲ) and G protein subunit(Gs?), and the interfering action of anisodamine. METHODS PMVEC was isolated and cultured from Wistar rat in vitro, radi- oligand binding assay and flow cytometer were used. RESULTS After incubation of LPS in vitro, the central F-actin of PMVEC depolymerized and its density decreased obviously , while the permeability of PMVEC monolyer increased significantly. Meanwhile it was found that LPS can induce the down regulation of 5-AR and Gsa protein level. Anisodamine can in- hibit these changes above. CONCLUSION LPS can directly cause the increase of permeability of PMVEC monolayer which relates to the depolymerization of central F-actin in PMVEC. Anisodamine may take part in regulating actin cytoskeleton of PMVEC and attenuate LPS-induced the increase of permeability of PMVEC monolaver by influencing two links of both ?-AR and Gsa protein

To study the cellular pathophysiological change in kidney IMCD cell under respiratory acid base disorders. 3% or 10% CO 2 was used to initiate chronic respiratory alkalosis(ALG) or chronic respiratory acidosis(ACG) of cultured rabbit IMCD cell. 5% CO 2 was used in control group(CG). Fluorescent probe was used to measure H + /K + exchange function of these cells. In situ hybridization and RT PCR were used to measure cH K ATPase mRNA expression of these cells.The results showed that H + /K + exchange in ACG was significantly higher than that in CG ( P 0 05). It is suggested that chronic respiratory acidosis could induce an increase in H + /K + exchange function and cH K ATPase mRNA expression. However, chronic respiratory alkalosis could induce decreases in H + /K + exchange function and cH K ATPase mRNA expression with no statistical significance, the reason of which calls for further study.

Objective To investigate the effects of tumor necrosis factor ? (TNF ?) on ? adrenoceptor (? AR) and ? AR related G protein coupled receptor kinases (GRKs) in rat pulmonary microvascular endothelial cells (PMVECs) as well as the interfering action of anisodamine. Methods Radio ligand binding assay was used to measure the maximal binding capacity (B max ) changes of ? AR in rat PMVECs after treatment with TNF ?. The effects of TNF ? and ? AR related GRKs expression at the protein level were observed by Western blotting. Results B max of ? AR in normal rat PMVECs was (5.58?0.31) fmol/10 5 cells. B max of ? AR in TNF ? group decreased significantly as compared with that in the normal control group, but no significant difference was found between the normal control group and TNF ?+anisodamine group. The expression of GRK2 in rat PMVECs was positive, but expression of GRK3, GRK5, and GRK6 were negative. The expression of GRK2 in TNF ? group and TNF ?+anisodamine group increased significantly as compared with that in the normal control group, but no significant difference was found between the TNF ? group and the TNF ?+anisodamine group. Conclusion GRK2 but not GRK3, GRK5, or GRK6 is expressed in rat PMVECs. The increased expression of GRK2 induced by TNF ? in rat PMVECs might promote the phosphorylation of ? AR, leading to ? AR internalization and decoupling with G protein, which might be the main mechanism of down regulation of ? AR induced by TNF ?. Anisodamine might inhibit the down regulation of ? AR through other mechanism instead of inhibiting the increase of GRK2 expression.

Objective: To establish the standard for quality control of Zhiganqing Granules (Radix et Rhizoma Rhei, Fructus Ligustri Lucidi, Rhizoma Alismatis, Pericarpium Citri Reticulatae, Polyporus Umbellatus, etc.) Methods: Fructus Ligustri Lucidi, Rhizoma Alismatis and Pericarpium Citri Reticulatae in the preparation were identified by TLC. The emodin in the preparation was determined by HPLC. A good linear range was shown at the concentration from 0.224?g to 0.732?g. The average recovery was 96.8% and the RSD was 1.14%.

AIM　To establish the fluorescent measurement technique for H+/K+ exchange in primary cultured IMCD cell monolayers of rabbit kidney. METHODS　To measure H+/K+ exchange in the primary cultured IMCD cell monolayers under different CO2 concentrations by the BCECF/AM fluorescent probe.RESULTS　［pH］i recovery rate after acid loaded in the respiratory acidosis group was 0.0620±0.012，and was significantly higher than that of the control group（0.0504±0.009，P<0.05）;which in the respiratory alkalosis group was 0.0476±0.007, there was no signifacant difference between ALG and CG （P>0.05）. CONCLUSION　The fluorescent measurements of H+/K+ exchange in primary cultured IMCD cell monolayers of rabbit kidney has many benefits which are convenient manipulation, stable and credit result, economy and safety , this method should further be popularized.

Objective To explore the effects of early glucocorticoids (GC) therapy on the outcome of patients with acute respiratory distress syndrome (ARDS). Methods The clinical data of all ARDS patients admitted from January 2008 to December 2011 in Chengdu Military General Hospital of Chinese PLA were retrospectively analyzed. The adult patients whose diagnosis was in accord to the Berlin ARDS diagnostic criteria published in 2012 were enrolled, and based on whether using glucocorticoid or not, they were divided into GC group and non-GC group. All the patients in GC group received low dosage of intravenous GC within 48 hours after the onset of ARDS, including different kinds of GC, methylprednisolone, dexamethasone and hydrocortisone (hydrocortisone dosage < 5 mg·kg-1·d-1, the dosage of former two kinds of GC being converted to that of hydrocortisone), and the therapeutic course of the two groups was 7 to 21 days. The patients in non-GC group received no GC therapy after the occurrence of ARDS. The duration of mechanical ventilation, the length of intensive care unit (ICU) stay and totally in hospital, medical cost and 28-day survival rate were compared between the two groups. Results One hundred and seventeen patients with ARDS were collected, including 56 cases (47.86%) in GC group and 61 cases (52.14%) in non-GC group. The duration of mechanical ventilation in GC group was significantly shorter than that in non-GC group [days:0 (0, 2.50) vs. 2.00 (0, 2.50), Z=2.015, P=0.044]. The 28-day survival rate in GC group was significantly higher than that in non-GC group [71.43%(40/56) vs. 50.82%(31/61),χ2=5.198, P=0.023]. There were no significant differences in the length of ICU stay [days:7.50 (2.00, 11.00) vs. 4.00 (1.00, 9.00), Z=1.879, P=0.060] and stay totally in hospital [days:16.00 (10.00, 27.75) vs. 15.00 (7.00, 28.00), Z=0.592, P=0.552] between GC group and non-GC group. However, the medical cost in non-GC group was significant lower than that in GC group [10 thousand Chinese yuan:3.15 (1.51, 5.78) vs. 4.39 (1.66, 10.88), Z=2.204, P=0.028]. Conclusion The early GC therapy may improve the outcome of patients with ARDS, especially beneficial to the 28-day survival rate.

AIM To establish the fluorescent measurement technique for H+ /K+ exchange in primary cultured IMCD cell monolayers of rabbit kidney. METHODS To measure H+ /K+ exchange in the primary cultured IMCD cell monolayers under different CO2 concentrations by the BCECF/AM fluorescent probe. RESULTS [pH], recovery rate after acid loaded in the respiratory acidosis group was 0.0620 ? 0.012, and was significantly higher than that of the control group (0.0504 ? 0.009, P 0.05). CONCLUSION The fluorescent measurements of H+ /K+ exchange in primary cultured IMCD cell monolayers of rabbit kidney has many benefits which are convenient manipulation, stable and credit result, economy and safety, this method should further be popularized.

AIM: To determine if aquaporin-1 (AQP-1) expression and function is influenced after lipopolysaccharide (LPS) stimulation in rat lung microvessel endothelial cells and to investigate the mechanisms of lung fluid abnormal metabolism in acute lung injury. METHODS: LPS at different concentrations (100 ?g/L, 1 mg/L or 10 mg/L) was used to stimulate cultivated rat lung microvessel endothelial cells in vitro at different stimulatory times (4 h, 12 h or 24 h), respectively. Tritium water permeation was conducted for determining the intracellular signal intensity in rat lung microvessel endothelial cells. The RT-PCR technique was applied for the assay of the expression of AQP-1 mRNA. RESULTS: The signal intensity of intracellular tritium water in the LPS stimulation group was less than that in normal control significantly. Compared with the normal control group (P