Abstract: A new selaginellin derivative named as selaginellin S (1) was isolated from the whole plants of Selaginella pulvinata (Hook. et Grev.) Maxim. (Selaginellaceae), together with a known one (selaginellin M, 2). Compounds 1 and 2 were separated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were determined on the basis of extensive spectroscopic analysis including IR, MS, 1D and 2D NMR experiments, as well as ECD calculations. Compound 1 is a key intermidiant in the biosynthesis pathway of selaginellins. Compound 2 is first reported in this plant.

AIM: To study the effect of activated protein C(APC) at different concentrations on apoptosis of human umbilical vein endothelial cells(HUVECs) induced by lipopolysaccharide(LPS).METHODS: The HUVECs were induced by LPS(1.0 mg/L) as apoptotic model that was administered by different concentration of APC(10 ?g/L or 50 ?g/L).Meanwhile,the control group and induced apoptosis group induced by LPS(1.0 mg/L) stimulation were also set up.The changes of cellular ultrastructures were observed under electron microscope.The DNA ladder and TUNEL fluorescent staining were measured in cells.Annexin-Ⅴ/PI double staining was used to measure the cell apoptosis rate by flow cytometry.Cell survival rate was measured by MTT assay.The proliferating cell nuclear antigen(PCNA) expression levels in cells were also measured by Western blotting to reflect the proliferation of the cells.RESULTS: There were significant apoptotic changes in the cells induced by LPS,but the apoptotic changes were reduced and apoptosis rates were decreased in the cells treated with APC.Meanwhile,cell survival rate and the protein levels of PCNA were increased after APC treatment,particularly at the concentration of 50 ?g/L,which showed difference when compared with those induced apoptosis group by LPS(P

The paper is to report the pharmacokinetics of sulfamethoxazole in healthy Han volunteers living at plain (PH) and native Han and Tibetan healthy volunteers living at high altitude (HNH and HNT). After healthy volunteers were administrated orally cotrimoxazole tablets, plasma concentration of sulfamethoxazole and metabolite N4-acetylsulfamethoxazole was determined by RP-HPLC, and plasma concentration-time data were analyzed by DAS 2.0 software to get the related pharmacokinetic parameters. The main pharmacokinetic parameters t(1/2) of sulfamethoxazole in PH, HNH and HNT were, respectively, 9.30 +/- 1.11, 10.99 +/- 1.23 and 10.44 +/- 1.05 h; tmax were 1.4 +/- 0.3, 2.0 +/- 1.1 and 1.8 +/- 0.4 h; Cmax were 94.42 +/- 15.26, 89.33 +/- 7.67 and 87.43 +/- 11.61 micro x mL(-1); AUC(0-t) were 1202.5 +/- 238.3, 1 434.7 +/- 193.9 and 1302.8 +/- 103.0 microg x h x mL(-1); AUC(0-infinity) were 1240.7 +/- 255.3, 1511.5 +/- 211.9 and 1363.9 +/- 116.5 microg x h x mL(-1); CL were 1.01 +/- 0.22, 0.81 +/- 0.12 and 0.89 +/- 0.08 L x h(-1) x kg(-1); V were 13.27 +/- 1.73, 12.81 +/- 2.15 and 13.28 +/- 1.20 L x kg(-1). Sulfamethoxazole pharmacokinetic parameters of HNH and HNT were significantly different from that of PH. The t(1/2) was significantly higher and the CL was significantly lower in HNH and HNT than that in PH, and the AUC(0-infinity) was significantly lower in HNT compared with HNH. This study found significant changes in the disposition of sulfamethoxazole under the special environment of high altitude hypoxia. This finding may provide some references for clinical rational application of sulfamethoxazole in HNH and HNT.

Objective To investigate the relationship between the activity of Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway in cerulein-induced pancreatic acinar cell line AR42J. Methods The in vivo model of AP was induced by cerulean treated pancreatic acinar cell line AR42J, then RPM and AG490 were given for intervention. Western blot was used to determine theexpressions of JAK1 and phosphorylation JAK1 ( P JAK1 ) , STAT1, PSTAT1 and TNFα, IL-1β, IL-6. The expressions of IL-6, IL-1 β, and TNFα mRNA were measured by RT-PCR. Survival rate of cells was evaluated by trypan blue stain. Results The relative expressions of JAK1, P JAK1, STAT1, P STAT1 and TNF-o, IL-1β, IL 6 without cerulean treatment were 0.09 ±0.04,0.14 ±0.08,0.21 ±0.09,0.12 ±0.12,0.10 ±0.02,0.08 ± 0.03,0.02 ± 0.02. After cerulean treatment, the expressions of abovementioned protein increased in a time-dependant manner, the expressions at 24h were 0.53 ± 0.09,0.53 ± 0.13,0.56 ± 0.09,0.55 ± 0.10,0.25 ± 0.04,0.25 ±0.09,0.27 ±0.07, which were significantly higher than those in the control group (P ＜0.05). 2 4 h after RPM and AG 4 9 0 inhibition, the expressions of TNF-α, IL-1 β, IL-6 proteins significantly decreased to 0.17 ± 0.03and 0.17 ± 0.01,0.15 ± 0.05 and 0.14 ± 0.07,0.19 ± 0.04 and 0.19 ± 0.05; their expressions of mRNA significantly decreased ( P ＜ 0.05 ). The cell survival rates in RPM and AG490 treatment group were (72.4 ± 11.2) %, (69.7 ± 9.8 ) %, and in cerulein-stimulated cells (42.2 ± 12.3 ) % ( P ＜ 0.05 ).Conclusions The JAK1/STAT1 signaling pathway was involved in pancreatic inflammatory response with cerulein stimulation. Early treatment with inhibitors to the JAK1/STAT1 signaling pathway might control the inflammatory response in acute pancreatitis.

Objective To investigate the role of connexin 43(Cx43)in the apoptosis of pancreatic cancer cell line BxPC and its possible mechanism.Methods pcDNA-Cx43,pcDNA-Cx43N,pcDNA3.0,siRNA-Cx43 and siRNA-NC were transfected into BxPC3 cells via liposome method.Cx43 protein and Cytochrome C(Cyt C)concentration was determined by Western blot,and the apoptosis was analyzed by Annexin V/PI binding assay.The mitechondria apoptosis pathway involved in Cx43 associated apoptosis was examined which contains the depolarization of mitechondrial membrane potential (MMP);fluorospectrophotometer was used to measure the activities of caspase-3 and caspase-9. Gap junction intercellular communication(GJIC) was determined by dye-transfer method.Results Cx43 protein expression increased after BxPC3 transfeetion,apoptosis rate increased from(6.35±0.43)%in empty vector transfection group to(14.29±1.24)%;after H202 treatment,apoptosis rate increased from(20.34±2.47)%to(31.27±2.56)%(P<0.05).Meanwhile,mitochondrial membrane potential was decreased,Cyt C was increasingly released from mitochondria,caspases activities were increased;after siRNA43 interference,apoptosis rate decreased from(7.42±0.47)% to(5.19±1.37)%,after H_2O_2 treatment,apoptosis rate decreased from (19.43±1.71)%to(11.67±1.97)%(P<0.05).Decreased mitochondrial membrane potential and Cyt C release were observed,caspases activities were decreased.GJIC of pcDNA-Cx 43 transfection group increased from 14.52±0.57 to 23.05±3.84.and it increased from 1.70 ±0.24 to 3.84 ±0.45 in the presence of β-GA(P<0.05).But the apoptosis rate was not significantly different.Conclusions Cx43 could promote BxPC3 apoptosis via mitochondrial apoptotic signal pathway,and the possible mechanism included signal pathway other than GJIC.

Objective To study the protective effects of activated protein C (AFC) on the apoptosis of endothelial cells induced by lipopolysaccharide (LPS) in order to clarify the mechanisms associated with the expression of some genes related to apoptosis. Method The human umbilical vein endothelial cells were incubated with LPS (1.0 μg/mL) for one hour to make the models of cell apoptosis, and then the different concentrations of AFC (10 ng/mL and 50 ng/mL) were added to the models of cell apoptosis as treatment group. Therefore, there were two groups, model group and APC treated group. The factors related with apoptosis such as P53, Bax, Bcl-2, and caspase-3 mRNA or protein level were measured by using RT-PCR and Western blotting. Results Compared with LPS stimulated cells, the expressions of P53, Bax and caspase-3 mRNA and levels of protein were decreased and the expression of Bcl-2 mRNA and protein level were increased in APC treated cells particularly in APC 50 ng/mL treated cells (P <0.05). Conclusions The APC inhibits the apoptosis of HUVECs induced by LPS via regulating the mitochondrial-dependent apoptosis pathway, and it may become a novel therapeutic agent for infection disease.

Objective To explore the relationship between the severity of experimental acute pancreatitis (AP) and the expression of Myc interacting zinc finger protein 1 (MIZ1) in rat, to evaluate the value of MIZ1 for severity assessment .Methods Acute necrotizing pancreatitis ( ANP) model was induced by retrograde injection of sodium taurocholate into the pancreatic duct , and normal saline was used in control group.The rats were sacrificed at 6, 24, 48 h, and the blood and pancreas tissue was collected , and serum amylase level , C reactive protein ( CRP ) , TNF-α, IL-6 and MIZ1 were determined by ELISA .Pancreatic tissue was routinely examined by pathologist , and the MIZ1 protein expression in pancreatic tissue was measured by immunohistochemistry and Western blot .Results The serum amylase levels of control group and ANP group at 6, 24, 48 h were (449 ±40), (578 ±25), (1 021 ±205), (971 ±143)U/L, and the levels of CRP were (123 ±23), (169 ±25), (226 ±34), (229 ±24)mg/L;and the levels of IL-6 were (20.16 ± 4.11), (38.60 ±12.05), (52.33 ±6.77), (44.83 ±4.30)ng/L;and the levels of TNF-αwere (55.33 ±3.32), (82.8 ±5.26), (120.66 ±16.00), (108.33 ±12.17)ng/L;and the levels of MIZ1 were (5.51 ± 0.34), (3.44 ±0.56), (2.11 ±0.11), (2.41 ±0.43) ng/L.The pathologic scores of pancreas were (1.83 ±0.75), (6.00 ±1.67), (8.16 ±2.70), (9.33 ±1.50), and the expressions of MIZ1 in pancreatic tissue were 0.81 ±0.05, 0.53 ±0.07, 0.31 ±0.06, 0.21 ±0.08.Except for amylase level of ANP 6h group, other parameters of ANP group were significantly different with those of control group , and the parameters of ANP 24, 48 h group were significantly different with that of ANP 6 h group ( P<0.01), but there was no significantly different between ANP 24 and 48 h group.MIZ1 expression was negatively correlated with serum amylase level , CRP, TNF-α, IL-6 and pathologic scores of pancreas , and the difference was statistically signific ant (P<0.01).Conclusions The decreasing expression of MIZ1 is closely correlated with the severity of AP , and may be a potential marker for prognosis evaluation .

BACKGROUND:Al oying is a convenient and effective method to alter the microstructure and control the corrosion behavior of magnesium al oy.
OBJECTIVE:To explore the in vitro and in vivo degradation of Mg-Nd-Zn-Zr al oy as a degradable medical biomaterial.
METHODS:(1) In vitro static immersion test:The immersion tests were carried out at (37.0±0.5) thermostatic bath. Six Mg-Nd-Zn-Zr al oy samples and six pure magnesium samples were immersed in the 250 mL simulated body fluid and vibrated without agitation during immersion. After 3, 7 and 30 days static immersion, the samples were taken out from the simulated body fluid. Then the in vitro corrosion properties were evaluated using scanning electron microscope and energy dispersive spectrometer analysis. (2) In vivo animal experiment:After bone tunnel was established in the left femur of adult New Zealand rabbits, the Mg-Nd-Zn-Zr al oy rods were embedded in the Mg-Nd-Zn-Zr al oy group, titanium al oy rods were embedded in the titanium al oy group, and only bone tunnel was established in the sham-operated group. At 1, 2, 4, 8 weeks after implantation, an X-ray of the implanted region was taken to determine the location and the degradation behavior℃in a of Mg-Nd-Zn-Zr al oy. At 4, 8 weeks after implantation, the corrosion product and its element composition were observed using scanning electron microscope with an energy dispersive spectroscopy system.

Objective To investigate the effect of antioxidants including PDTC on pancreatic fibrosis of rats with chronic pancreatitis.Methods The rats were randomly divided into 5 groups including control group, CP group, PDTC treatment group, vitamin E treatment group and vitamin C treatment group.The CP model was in ducad by using intraperitoneal injection of DETC (750 mg/kg), twice a week.The control group received no treatment.After DETC injection, the treatment groups received an intraperitoneal injection of PDTC (100 mg/kg), vitamin E (15 mg/kg), vitamin C (15 mg/kg), respectively.Rats were sacrificed at 90 min, 24 h, 48 h, 72 h, 2 w, 3 w, 4 w, 6 w after first injection of DETC.Pancreatic tissue was taken for routine pathological examination.The activity of SOD, GSH-PX and MDA content were detected by spectrophotometric ratio method.α-SMA, desmin collagen Ⅰ, Ⅲ, TGF-β1, FN were detected by immunohistochemical assay.The expression of TGF-β1, FN mRNA was measured by RT-PCR.Results At 6w, the fibrosis and the parameters for damage of the pancreas in the three treatment groups were significantly better than that in CP group (P ＜0.01), the vacuolar degeneration index in vitamin E group and vitamin C group was also better than that in CP group (P ＜0.01).From the 2nd week, the activity of SOD, GSH PX in PDTC group, Vit C group and Vit E group was higher than that in CP group, while the MDA activity was lower than that in CP group, and the difference was statistically significant (P ＜ 0.01 or P ＜ 0.05).No significant difference was found among the three treatment groups.The mRNA levels of TGF-β1 and FN of the treatment groups were lower than those of CP group (P ＜0.05 or P ＜0.01), but higher than those of the control group (P ＜ 0.05).There was no significant difference among the three treatment groups (P ＞ 0.05).Conclusions PDTC and the other antioxidants can reduce oxygen free radicals by increasing the activity of SOD,suppressing the activation of PSCs, reducing the secretion of TGF-β1, Collagen Ⅰ , Ⅲ, FN and eventually inhibit the progress of pancreatic fibrosis.

Objective To optimize the best subculture cycle through studying the influence of subculture time on tube plantlet growth of Dendrobium huoshanense and the change of medium composition.Methods Height,tiller,fresh weight,and chlorophyll content of the tube plantlet and pH value,water content,and sugar content of the medium were measured after different cycles of the subculture,the cost of culture medium for subculture was calculated as well.Results The height of the tube plantlet increase 282.86%,the tiller increase by 3.5 times,fresh weight reaches its maximum,chlorophyll content of the tube plantlet almost reaches its maximum after 40 d subculture;while water content and sugar content of the medium are decreased to the lowest point,pH value of medium(