Objective To investigate the effects of penehyclidine hydrochloride (PHCD) pretreatment on hypoxia-inducible factor-1alpha (HIF-1α) expression in lung tissues in rats with acute lung injury (ALI) induced by lipopolysaccharide (LPS).Methods One hundred and twenty female Sprague-Dawley rats,weighing 180-220g,were randomly divided into 3 groups (n =40 each):control group (group C),ALI group and PHCD pretreatment group (group P).ALI was induced by intraperitoneal LPS 5 mg/kg in ALI and P groups.The equal volume of normal saline was given intraperitoneally in group C.Group P received intraperitoneal PHCD 2 mg/kg at 30 min before LPS administration.Eight rats in each group were chosen at 2,4,8 and 24 h after LPS administration,and the lung was removed for determination of HIF-1α mRNA expression in lung tissues by RT-PCR.Eight rats in each group were chosen at 6 h after LPS administration,and the lung was removed for determination of W/D lung weight ratio and IL-6 content (by ELISA) and for microscopic examination.Results The W/D lung weight ratio and IL-6 content at 6 h after LPS administration and HIF-1α mRNA expression at each time point after LPS administration were significantly higher in groups P and ALI than in group C (P ＜ 0.05),and lower in group P than in group ALI(P ＜ 0.05).The pathological damage was significantly ameliorated in group P as compared with group ALI.Conclusion PHCD can reduce LPS-induced ALI through down-regulation of HIF-1α expression in lung tissues and inhibition of the inflammatory response in rats.

Objective To evaluate the safety and feasibility of hepatic resection for huge primary liver carcinoma (PLC). Methods 216 cases of huge PLCs(mean diameter of 14.2cm) were resected. The hepatectomies were performed under intermittent occlusion of hepatic inflow. Results All 216 cases were successfully resected. The mean time of occlusion of hepatic inflow was 19min, the mean blood loss was 743 ml. No serious complications occurred, and only seven patients died of hepatic failure and upper gastrointestinal haemorrhage postoperatively in this series. Conclusions Although resection of huge PLC is quite difficult, but if suitable surgical techique and perioperative management are adopted ,it is safe and feasible .

Objective To investigate the effects of huaier granules on invasion and metastasis of colorectal cancer SW480 cells in vitro, and explore the basic mechanism. Methods The appropriate concentration and duration of huaier granules promoting SW480 cell apoptosis were determined by SubG1 method. Wound healing assay and transwell assay were used to observe the effect of huaier granules on SW480 cell invasion and metastasis. The changes of E-cadherin, twist, snail and vimentin at protein and mRNA levels were examined by Western blotting and Real-Time PCR. Results After treatment with huaier granules at 3. 0 g·L-1 for 36 h, apoptosis of SW480 cells was most significant, and wound healing assay revealed that relative mobility was (31. 36±2. 39)%, compared with (61. 11±1. 09)% in control group (P<0. 01). Number of invaded cells per field of view was (129±12) in treatment group and (354±20) in control group (P<0. 01). After treatment with huaier granules at 3. 0 g·L-1 for 36 h, protein and mRNA levels of E-cadherin were increased, while those of twist, snail and vimentin were decreased. Conclusion Huaier granules can inhibit invasion and metastasis of colorectal cancer SW480 cells in vitro through effectively depressing epithelial-mesenchymal transition.

Objective To investigate the effect of penehyclidine hydrochloride on endotoxemiainduced intestinal injury in neonatal rats.Methods Ninety healthy neonatal Sprague-Dawley rats,aged 7days,weighing 16-20 g,were randomly divided into 3 groups (n =30 each) using a random number table:control group (group C),lipopolysaccharide (LPS) group,and penehyclidine hydrochloride group (group P).Endotoxemia was induced by intraperitoneal LPS 5 mg/kg in LPS and P groups.In group P,penehyclidine 2 mg/kg was injected intraperitoneally at 30 min before and after LPS injection.The equal volume of normal saline was given in group C.At 2,6 and 12 h after LPS or normal saline administration,10 rats in each group were randomly chosen and sacrificed.The ileum was removed to detect wet/dry lung weight ratio (W/D ratio) and contents of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),hypoxia-inducible factor 1 alpha (HIF-1α) and glutamine (Gln),and for examination of the pathologic changes of the ileum with light microscope.Results Compared with group C,the W/D ratio and contents of TNF-α,IL-6 and HIF-1α were significantly increased,and the content of Gln was decreased at each time point in LPS and P groups.Compared with group LPS,the W/D ratio and contents of TNF-α,IL-6 and HIF-1α were significantly decreased,the content of Gin was increased at each time point,and the pathologic changes of the ileum were mitigated in group P.Conclusion Penehyclidine hydrochloride can reduce endotoxemia-induced intestinal injury in the neonatal rats,and down-regulated expression of HIF-1α,up-regulated expression of Gln and attenuated inflammatory responses may be involved in the mechanism.

Aim To determine the neuroprotective effect of sulfated pachymaran (SP)on MPTP-induced
mouse model.Method ICR mice were randomly di-vided into control group,MPTP group and SP treatment group (50,1 00,1 50 mg·kg -1 ,ip).After 1 7 days, the activities of endogenous antioxidants (SOD,GSH-Px,CAT),antisuperoxide anion,hydrogen peroxide and MDA content in the midbrain and cortex were as-sayed.Results The results proved that SP significant-ly reduced the content of MDA and H2 O2 ,regulated
the activities of antioxidant enzyme and increased the activity of antisuperoxide anion.Conclusion All these effects indicate that SP is a potential neuroprotective a-gent and its neuroprotective effects are achieved in the MPTP mouse model.

DNA damage response (DDR) in different cell cycle status of human peripheral blood lymphocytes (PBLs) and the role of H2AX in DDR were investigated. The PBLs were stimulated into cell cycle with phytohemagglutinin (PHA). The apoptotic ratio and the phosphorylation H2AX (S139) were flow cytometrically measured in resting and proliferating PBLs after treatment with camptothecin (CPT) or X-ray. The expressions of γH2AX, Bcl-2, caspase-3 and caspase-9 were detected by Western blotting. DDR in 293T cells was detected after H2AX was silenced by RNAi method. Our results showed that DNA double strand breaks (DSBs) were both induced in quiescent and proliferating PBLs after CPT or X-ray treatment. The phosphorylation of H2AX and apoptosis were more sensitive in proliferating PBLs compared with quiescent lymphocytes (P<0.05). The expression levels of anti-apoptotic proteins Bcl-2 were reduced and cleaved caspase-3 and caspase-9 were increased. No significant changes were observed in CPT-induced apoptosis in 293T cells between H2AX knocking down group and controls. It was concluded that proliferating PBLs were more vulnerable to DNA damage compared to non-stimulated lymphocytes and had higher apoptosis rates. γH2AX may only serve as a marker of DNA damage but exert no effect on apoptosis regulation.

Objective:To evaluate the relationship between Na + leak current channel (NALCN) in hippocampal dentate gyrus (DG) region and cognitive function in mice. Methods:Thirty-nine male wild type C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were studied.Three mice were sacrificed to verify the expression of NALCN co-localized with neuronal nuclear antigen (NeuN) in the hippocampal DG region through immunofluorescence technique.The remaining 36 mice were divided into 2 groups ( n=18 each) by the random number table method: control group (group C) and NALCN gene knockout group (group KO). NALCN-shRNA virus was injected in hippocampal DG region in group KO, and scrambled-shRNA virus was injected in group C. Three weeks after virus injection, behavioral tests (Y maze test and open field test) were performed, then the animals were sacrificed, and the hippocampal tissues were removed for determination of the expression of NALCN protein and mRNA using Western blot and real-time polymerase chain reaction. Results:NALCN and NeuN colocalized a lot on the same neuron in the hippocampal DG region of mice, and NALCN was widely expressed in the hippocampal DG region.Compared with group C, the expression of NALCN protein and mRNA was significantly down-regulated, the times of entering the new arm were reduced, the duration of staying at the new arm was shortened ( P<0.05), and no significant change was found in the parameters mentioned above in the open field test in group KO ( P>0.05). Conclusions:NALCN in the hippocampal DG region is involved in the regulation of cognitive function in mice, and the down-regulation of NALCN may lead to cognitive decline.

AIM:We investigated how AT 1-R stimulated by mechanical stresses induces cardiac fibrosis .METHODS:We produced in vivo cardiac pressure overload model in angiotensinogen knockout ( ATG-/-) mice and in vitro mechanically-stretched cell model in cultured neonatal cardiac cells of ATG-/-mice both lack the participation of Ang II .RESULTS: Pressure overload for 4 weeks in ATG-/-mice induced myocardial hypertrophy accompanied by the significant interstitial fibrosis , however , the TGF-β, a key regulatory factor of fibrosis, was not significantly increased in these ATG-/-mice.Meanwhile, the inhibitor for AT1-R significantly inhibited mechani-cal stress-induced cardiac fibrosis in these ATG-/-models whereas inhibition of TGF-βdid not.CONCLUSION:The results showed that mechanical stress-induced fibrotic responses through AT 1-R required the phosphorylation of Smad 2 but not the involvement of TGF-β.

DNA damage response (DDR) in different cell cycle starus of human peripheral blood lymphocytes (PBLs) and the role of H2AX in DDR were investigated.The PBLs were stimulated into cell cycle with phytohemagglutinin (PHA).The apoptotic ratio and the phosphorylation H2AX (S139)were flow cytometrically measured in resting and proliferating PBLs after treatment with camptothecin (CPT) or X-ray.The expressions of γH2AX,Bcl-2,caspase-3 and caspase-9 were detected by Western blotting.DDR in 293T cells was detected after H2AX was silenced by RNAi method.Our results showed that DNA double strand breaks (DSBs) were both induced in quiescent and proliferating PBLs after CPT or X-ray treatment.The phosphorylation of H2AX and apoptosis were more sensitive in proliferating PBLs compared with quiescent lymphocytes (P＜0.05).The expression levels of anti-apoptotic proteins Bcl-2 were reduced and cleaved caspase-3 and caspase-9 were increased.No significant changes were observed in CPT-induced apoptosis in 293T cells between H2AX knocking down group and controls.It was concluded that proliferating PBLs were more vulnerable to DNA damage compared to non-stimulated lymphocytes and had higher apoptosis rates.γH2AX may only serve as a marker of DNA damage but exert no effect on apoptosis regulation.