Aim To observe the effects of theaflavin on intracellular calcium concentration(i) level in rat ventricular myocytes and discuss the possible mechanisms.Methods The effects of theaflavin oni were investigated in rat ventricular myocytes.i was detected by laser confocal microscopy and represented by relative fluorescent intensity(FI-FI0)/FI0,%;FI0:control;FI:administration of drugs).Results ① Theaflavin(10,20,40 ?mol?L-1) had no effect on the i of ventricular myocytes in normal Tyrode′s solution.However,it reduced the i of ventricular myocytes in simulated ischemia solution in a concentration-dependent manner.② Pretreatment with Bay k8644(0.5 ?mol?L-1) mostly abolished the effects of theaflavin(20 ?mol?L-1) in simulated ischemia solution.③ Theaflavin(20 ?mol?L-1) markedly inhibited the low concentration of ryanodine-induced i increase in Ca2+-free Tyrode′s solution.④ When the propagating waves of elevated i(Ca2+ waves) were produced by increasing extracellular Ca2+ concentration from 1 mmol?L-1to 10 mmol?L-1,theaflavin(20 ?mol?L-1) could block the propagating waves of elevated i(Ca2+ waves),reduce the frequency and duration of propagating waves,and reduce i as well.Conclusion Theaflavin may reduce the i in isolated rat ventricular myocytes via inhibiting Ca2+ influx by voltage-dependent Ca2+ channel and alleviating Ca2+ release from sarcoplasmic reticulum(SR).

Aim To observe the influences of ginsenoside Rg1 on the spontaneous contraction of small intestine smooth muscle of rabbits in vitro and explore the mechanism.Methods With the isothermal perfusion of small intestine in vitro,the influences of ginsenoside Rg1 on the spontaneous contraction of small intestine was observed and the mechanism of ginsenoside Rg1 was studied.Results Ginsenoside Rg1 reduced the amplitude of contraction of small intestine smooth muscle in rabbits in a dose-depended manner.Bay K8644 and nitro-L-arginine methylester(L-NAME)could completely block the inhibition of ginsenoside Rg1 on the contraction of small intestine smooth muscle.Ginsenoside Rg1 inhibited the intracellular calcium-depended contraction induced by rynodine in the Ca2+ free Tyrode's solution.Conclusions Ginsenoside Rg1 inhibits the contraction of small intestine smooth muscle of rabbits in vitro.The mechanism may be related to increase NO concentration in small intestine smooth muscle so that it inhibits extracellular Ca2+ inflowing via cell membrane and intracellular Ca2+ releasing via sarcoplasmic reticulum.

Aim To investigate the effect of ginsenoside Rg1 on gut injury following intestinal ischemia reperfusion in rats and the possible mechanism.Methods By using rat model of intestinal I/R injury, 30 male SD rats were randomly divided into 3 groups(n =10 in each group):sham-operation group, I/R group(control group)and ginsenoside Rg1 group(treatment group).The contents of tumor necrosis factor α(TNF-α), interleukin-6(IL-6), malondialdehyde(MDA), the activity of superoxide dismutase(SOD)in intestinal mucosa were measured respectively.Chiu's count was used to assess the changes in intestinal pathological morphology.Results TNF-α, IL-6, MDA contents and the intestinal injury score in control group were significantly increased compared to those in sham-operation group, while SOD contents in control group were significantly decreased compared to sham-operation group.Inversely, TNF-α, IL-6, MDA contents and the intestinal injury score in treatment group were significantly decreased compared to those in control group, while SOD contents in treatment group were significantly increased compared to control group.Conclusion Pretreatment with ginsenoside Rg1 has protective effect on intestinal ischemia-reperfusion injury in rats, which may be attributed to decreased contents of TNF-α, IL-6, MDA and increased levels of SOD.

Aim To observe the cardioprotection of chronic intermittent hypobaric hypoxia (CIHH) against ischemia/reperfusion injury in adult and young rats.Methods Adult and postnatal male Sprague-Dawley rats were divided randomly into four groups:control 28-day group (CON28),control 42-day group (CON42), CIHH 28-day treatment group (CIHH28), and CIHH 42-day treatment group (CIHH42). CIHH animals with maternal rats were put into a hypobaric chamber 2 days before birth to get 28 days and 42 days CIHH mimicking 3000 m altitude (P_B=525 mmHg,P_(O_2)=108.8 mmHg), 5 h daily, respectively.The control animals were kept in the same environment as CIHH rats with free access to water and food except hypoxic exposure. The isolated hearts were perfused in the Langendorff apparatus, undergoing 30 min global ischemia and 60 min reperfusion.Cardiac function was recorded continuously during the whole experiment. Parameters of left ventricular function included left ventricular developing pressure (LVDP), left ventricular end-diastolic pressure (LVEDP), maximal positive (+LVdp/dt) and negative (-LVdp/dt) velocity of left ventricular pressure, coronary flow (CF) and heart rate(HR).Results ① For adult rats, there was no significant difference in the parameters of left ventricular function between CIHH28 and CON28 groups. However, the recovery of cardiac function in CIHH42 rats was much better than that in CON42, including LVDP, LVEDP, ±LVdp/dt and CF (P<0.05). ② For young rats, the basic coronary flow (CF) in CIHH rats was significant higher than that in CON rats, while other parameters of cardiac function didn't change. The recovery of cardiac function in CIHH rats was much better than that in CON rats, including LVDP, LVEDP,±LVdp/dt and CF (P<0.05).Conclusion CIHH confers cardioprotection against ischemia/reperfusion injury in rat cardiomyocytes, which is predominant in CIHH42 group and significantly affected by the age of animals. Cardioprotective effects produce more easily in young rats by CIHH.

Aim To investigate the effect of chronic intermittent hypobaric hypoxia ( CIHH) on the paeonol induced vasomotion of isolated rat ’ s thoracic aorta rings and its underlying mechanisms. Methods Spra-gue-Dawlay ( SD ) rats were randomly divided into 2 groups: control group ( CON ) and CIHH treatment group ( CIHH) . CIHH rats were exposed to hypoxia in a hypobaric chamber simulating 5 000 m altitude, 6 hours daily for 28 days. CON rats lived in the same en-vironment as CIHH animals except hypoxia. Organ bath technique was used to observe the effect of pae-onol on isolated thoracic aorta rings of rats. Results There were no significant differences of noradrenaline ( NE )- and KCl-induced contraction in thoracic aorta rings among CIHH and CON rats;CIHH enhanced va-sodilative effects of paeonol on isolated thoracic aorta rings of rats; the vasodilative effects on CIHH rats could be partly decreased by β-receptor blocker prop-ranolol,ATP-sensitive potassium channel ( KATP ) bloc-ker glibenclamide and NO synthase inhibitor L-NAME. Paeonol significantly inhibited NE-induced intracellular and extracellular calcium-dependent contraction in CIHH rats. Paeonol didn ’ t inhibit NE-induced con-traction by intracellular calcium release and its inhibi-tory effect couldn ’ t be blocked by glibenclamide in CON. Vasodilative effects of paeonol couldn ’ t be re-versed by indomethacin, a cyclooxygenase inhibitor, in CIHH and CON rats. Conclusion CIHH significantly enhances vasodilative effects of paeonol on isolated tho-racic aorta rings of rats. Besides promoting the signa-ling pathway of paeonol in CON, CIHH significantly enhances vasodilative effects of paeonol via activating KATP and inhibiting Ca2+ release from sarcoplasmic re-ticulum.

OBJECTIVE:To investigate the new management model of the hospital pharmacy established by application of vender managed inventory (VMI) model. METHODS:Concerned about the implementation of VMI model,its effects and prob-lems,new management model of hospital pharmacy were described after the hospital cooperated with two suppliers. The hospital is mainly in charge of the management and application of pharmaceutical staff and equipment,drug capital flow,and the selection and optimization of drug use list;focus on the training of pharmaceutical talents;improve drug quality management,pharmaceuti-cal administration,pharmaceutical care and rational drug use. Two suppliers are mainly in charge of drug logistics,and the alloca-tion of servicer,software and hardware(as automatic medicine dispensing machine);share logistic cost of supply chain. Drug in-formation flow was established by both hospital and supplier. RESULTS:After cooperation,pharmaceutical staff,related equip-ment and inventory cost(floating capital decreased by 10 million yuan each month)were decreased,and the efficiency of distribu-tion improved;the error rate of drug dispensing in outpatient pharmacy were decreased (decreasing from 0.39‰ to 0.12‰) after the establishment of automatic pharmacy. The establishment of early warning system for abnormal patients flowrate in pharmacy window shortened the time of getting medicine,and improved service level and satisfactory degree of patients (increasing from 84.91% to 93.62%). CONCLUSIONS:New management model of hospital pharmacy based on VMI model provides reference for public hospital reform and hospital-enterprise cooperation under New Medical Reform.

Obej ctive Abnormal proliferation of T cells plays an important role in the development of autoimmune diseases. The article aimed to study the inhibitory effect of small molecule compound BD691 on T cell proliferation and its mechanism. Methods Human peripheral blood T-lymphocytes were isolated and purified by the immunomagnetic microbeads,then T cells were ac-tivated with anti-CD3/CD28 mAbs or alloantigen.The inhibitory effect of BD691 on activated T cell proliferation, the cytotoxic effect BD891 on resting T cells and the expression of activated T cells marker CD25 were measured by flow cytometry.Furthermore, ELISA was used to detect the secretion of cytokines associated with T cell differentiation. Results BD691 significantly inhibited the prolif-eration of T cells being stimulated by anti-CD3/CD28 mAb or alloantigen in a dose-dependent manner, and IC50 values are (8.5 ± 1.5)μmol/L and (7.2 ±1.3)μmol/L, respectively.However, BD691 had no obvious cytotoxic effects on resting T cells and periph-eral blood mononuclear cells, even at a high concentration ( up to 100μmol/L) .In T cells which were not activated by anti-CD3/CD28 mAb, the percentage of CD25+T cells is only 1.6%of the total cells, while the number increased to 68% after activating treatment.Mean-while, in T cells which were activated by 0, 3.3, 10, 30μmol/L BD691, no obvious change of CD25 expression were observed, while immunosuppressant FK506 (0.1μmol/L) significantly decreased the expression of CD25 +T cells (14.9%).In unactivated T cells, 95.6%cells were at G0/G1 phase, while after activation, the percentage of cells at G0/G1 phase reduced to 57.7%.In addition, BD691 inhibited the secretion of IFN-γ, IL-6 and IL-17 in activated T cells, but had no effects on the secretion of IL-2, IL-4 and IL-10. Co nclusion BD691 exerts no effects on T cell activation, but it inhibits T cell proliferation by inducing T cell cycling arrest at G0/G1 phase.Moreover, BD691 inhibits the secretion of key cytokines (such as IFN-γ, IL-6, IL-17) closely related to the differ-entiation of Th1 and Th17 cells.The results suggest that BD 691 is a potential lead compound to develop a new immunosuppressant for the inhibition of abnormal proliferation and differentiation of T cells.

ObjectiveTo study the biological characterization and the genetic background of circulating CA16 strains in mainland of China for the purpose of CA16 vaccine development in the future.MethodsCA16 strains were isolated from throat swabs of patients with hand-foot-mouth disease and identified by neutralization assay and RT-PCR.The genotype of these isolates were determined by sequence alignment and phylogenetic analysis of VP1 gene.The proliferation dynamics and the plaque morphology were observed when propagated in Vero cells.The pathogenicity of these CA16 isolates was evaluated by challenging newborn mice.ResultsIn this study,six CA16 circulating isolates,BJ-1-6 were obtained.The RT-PCR products were 150 bp amplified with the general enterovirus primers and 210 bp with CA16 primers respectively,which cannot be amplified by EV71 primers.Additionally,these isolates were identified to display some obvious proliferation dynamics and plaque morphology when propagated for 96 h in Vero cells.The diameter of plaques were about 1.5 to 2 mm for BJ-1,BJ-2,BJ-4,BJ-6,4-5 mm for BJ-3 and 3 mm for BJ5,the plaques were regular except BJ-3.All the six isolates can be neutralized by the convalescent serum of patient infected with CA16.The virus titer of different isolates propagated for five passages in Vero cells was 7.0LgCCID50/ml.The sequence alignment of VP1 gene demonstrated that the genotypes of BJ-2,BJ-4,BJ5 were C1 and BJ-1,BJ-3,B J-6 were C,3 comparatively.The genetic distance of the VPI gene from theseisolates suggested that they were highly genetic identity with the homology of 90％ in nucleotide and 99％ in dedicated amino acid respectively.However,a distinctive difference in pathogenic ability in neonatal mice was found that the suckling mice challenged with BJ-3 & BJ-5 were paralyzed 4-5 d and dead 6-7d postchallenge,compared with the control group without any abnormality in the during of 14 d.ConclusionThe circulating CA16 isolates in China have different biological characteristics,different pathogenic ability and similar genetic backgrounds,which is helpful for the development of a CA16 vaccine in the future.

Objective To investigate the effect of sensory and motor nerve homogenates at different concentrations on the proliferation and osteogenesis differentiation of bone marrow mesenchymal stem cells (BMSCs) in rats.Methods The saphenous nerve and the muscle branch of the sciatic nerve in rats were extracted surgically as sensory and motor nerve tissues,respectively.The primary nerve homogenates (10 mg/mL) were prepared as per 10 mg tissue with 1 mL osteoblast inducing conditional media,and 10 times diluted after filtration purification to prepare sensory and motor nerve homogenates at concentration gradients of 1.0,0.1,0.01,0.001 and 0.0001 mg/mL.Cultivation GFP ± rat pups BMSCs in vitro were trained to P3 generation.The experiment was carried out in 3 groups.The sensory and motor nerve homogenates of 500 μL at the above 6 concentration gradients were added during cultivation respectively in the sensory nerve group (n =18) and the motor nerve group(n =18) while 500 μL of osteoblast inducing conditional media was added in the control group(n =3).Cell proliferation quantity detection and alkaline phosphatase (ALP) staining were used to assess the proliferation and differentiation of BMSCs after 14 days.Results According to the results of CCK-8,the cellular absorbance values at concentrations of 1.0 and 0.1 mg/mL homogenate in the sensory nerve group (1.957 ±0.065 and 1.751±0.073) were significantly greater than in the control group (1.145±0.087) while the cellular absorbance value at concentration of 10.0 mg/mL homogenate in the motor nerve group (0.304 ± 0.619) was significandy smaller than in the control group (1.145 ± 0.087) (P ＜ 0.05).According to the ALP staining,the amounts of cellular calcium nodules in the sensory and motor nerve groups (2.667 ± 0.816 and 3.000 ± 0.632,respectively) were significantly smaller than in the control group (11.833 ± 1.471) (P ＜ 0.05).Conclusion Sensory nerve homogenate is different from motor nerve homogenate in that it may promote proliferation of BMSCs and inhibit osteogenesis differentiation of BMSCs in a certain rage of concentrations.

Objective Benzothiazole derivative BD960 has immunosuppressive activity after cell -based assays for high-throughput screening.The paper aimed to investigate the involved mechanism of BD960 on T cell proliferation. Methods Human peripheral blood T-lymphocytes were isolated and purified by the immunomagnetic microbeads.Then the T cells were activated by anti-CD3/anti-CD28 mAbs or alloantigen.The effect of BD960 on activa-ted T cell proliferation, the cytotoxic effect BD960 on resting T cells and the expression of activated T cells marker CD25 were measured by flow cytometer.Cytokine levels, including IL-2, IL-4, IL-6, IL-10, IL-17A and IFN-γ, were determined by ELISA. Results BD960 significantly inhibited the proliferation of T cells stimulated by anti-CD3/anti-CD28 mAb or alloantigen in a dose-dependent manner.The IC50 value is (2.3 ±0.3)μmol/L or (2.5 ±0.3)μmol/L, respectively.Moreover, BD960 had no obvious cytotoxic effects on rest-ing T cells and peripheral blood mononuclear cells, even at a high concentration ( up to 100μmol/L) .The ratio of CD25 expression on T cell was 69.7%after stimulated by Anti-CD3/CD28 mAbs with 72 h, the concentration (0.625、2.5、10)μmol/L of BD960 also had no potent effects on the ratio, but 0.1μmol/L FK506 could inhibit CD25 expression as low as 9.4%.The G0/G1 phase of activated T cells was 58.5%after stimulated by BD960 with 96 h.BD960 could induce cell cycle arrest at the G0/G1 phase in activated T cells with the increase of concentration and RAPA in the concentration of 0.1 μmol/L was 91.5%.In addition, BD960 (0.625、2.5、10)μmol/L could inhibit the secretion of IFN-γ, IL-6 and IL-17 in activated T cells with the increase of concentration, without any effects on the secretion of IL-2, IL-4 and IL-10. Conclusion BD960 not only exerts the inhibition on the late stage of T cell activation of cell proliferation but also inhibits the secretion of inflammatory cytokines, such as IL-6, IL-17 and IFN-γ, while the mechanism of BD960 on T cell proliferation was not the same as FK506.As a result, BD960 has the potential to be the lead compound to develop a new immunosuppressant.