Objective:To determine the mechanism of sunitinib-induced autophagy in renal cell carci-noma cells.Methods:MTS assay was applied to detect the cell viability alteration under the treatment of sunitinib (2,8 μmol /L).The sunitinib-induced autophagy as well as cell apoptosis was measured and compared after knocking down autophagy-related protein Beclin1 and microtubule associated protein 1 light chain 3 fusion protein (LC3)by RNA interference.The transmission electron microscope was used to observe the formation of autophagosomes in ACHN cells.The fluorescence microscope was used to mo-nitor distribution and aggregation of endogenous LC3-Ⅱ.The expressions of protein such as LC3-Ⅱ,the autophagic regulation molecules protein kinase B /mammalian target of rapamycin (Akt/mTOR)and the symbol of apoptosis poly ADP-ribose polymerase (PARP)were capable to be detected by immunoblotting assay.Results:Sunitinib was able to significantly trigger cell viability loss in the renal carcinoma cell ACHN,which was both in a concentration-dependent and time-dependent manner (P <0.05 ).After reducing the autophagy by knocking down Beclin1 and LC3,the number of cleavage of PARP was in-creased remarkably,whereas there was nearly not any cleavage in the mock group.By the transmission electron microscope,there were more autophagic vacuoles in ACHN cells after being administrated with sunitininb compared with the control.And the nuclear-to-cytosol translocation as well as aggregation of LC3-Ⅱ was presented after sunitinib treatment by the fluorescence microscope,which was the proof of the enhanced autophagy.According to the immunoblotting,sunitinib was able to increase the accumula-tion of LC3-Ⅱ.At the same time,the result of sunitinib combined with chloroquine,a drug which blocked the fusion of autophagosomes and lysosomes,demonstrated that the increasing amount of LC3-Ⅱwas due to the enhanced autophagy flux by sunitinib treatment in ACHN cells.However,phosphorylation of Akt as well as mTOR was decreased at the same time.The rapamycin (mTOR inhibitor)or knocking down Akt subunits could change the sunitinib-induced LC3-Ⅱ accumulation,whereas overexpression of Akt subunits decreased the autophagic flux,indicating that Akt/mTOR was the target of sunitinib in auto-phagy.Conclusion:Sunitinib induced autophagy via suppressing Akt/mTOR pathway,and the auto-phagy was involved in apopotosis.

OBJECTIVE:To establish a method for analyzing the volatile components in Cuscuta chinensis,and compare the difference of the volatile components in C. chinensis. METHODS:HS-SPME-GC-MS was adopted:sampling amount was 1.0 g, extracting fibers was 65 μm PDMS/DVB,equilibrium temperature was 120 ℃,equilibrium time was 15 min,extraction time was 30 min,resolution time was 3 min;GC conditions:the column was HP-5MS quartz capillary column,programmed temperature, inlet temperature was 230 ℃,carrier gas was high purity helium,the flow rate was 1.0 ml/min,splitless injection;MS condi-tions:ion source was electron ionization,temperature was 230 ℃,quadrupole temperature was 150 ℃,electron energy was 70 eV,photomultiplier tube voltage was 1.2 kV,the interface temperature was 280 ℃,and scanning range was m/z 35-550. Com-bined with the qualitative analysis for volatile components of C. chinensis from different habitats by HP ChemStation,the relative content was calculated by peak area normalization,and the data was analyzed by principal component analysis and cluster analysis. RESULTS:Totally 52 components were identified,9 of which were the common components in C. chinensis,namely leaf alcohol, 1-octene-3-ol,3-octanol,malt alcohol,diethyl phthalate,caryophyllene,nonaldehyde,octanol and palmitic acid. sample 1,2,3 were clustered into a group,then clustered with 4,5,6 into a group,sample 7,8,9 was clustered into a group,then clustered with 10,11,12 into a group,and sample 13,14,15 clustered into a group individually. CONCLUSIONS:The method is stable and reliable,and suitable for the rapid analysis of volatile components in C. chinensis;and differences of volatile components in C. chinensis from diflerent habitats are discernible.

Objective To investigate the relationship between the matrix metalloproteinase-3(MMP-3)and the stability of carotid artery plaque,and explore MMP-3's prediction role on the attack and relapse of acute ischemic cerebrovascular events.Methods 100 patients with the first ever acute cerebral infarction,100 patients with chronic cerebral circulation insufficiency(CCCI)and 40 persons without cerebrovascular diseases were enrolled in this study.According to the carotid ultrasound examination,100 cerebral infarction patients were divided into three subgroup: unstable plaque group(45 patients,mixed plaque,soft plaque),stable plaque group(35 patients,plaque Group)and endometrial coarse group(25patients).Matrix metalloproteinase-3(MMP-3)levels of all the subjects were measured by enzyme-linked immunosorbent assay(as basal level).All the subjects were followed up for one year to observe cerebral infarction events.Serum MMP-3 levels of each group,and the basic serum MMP-3 levels were compared among patients who were attacked or relapsed cerebral ischemic with those who had not been attack cerebral ischemic during this period of time.Results 5 patients in the cerebral infarction group had relapse (5%),2 patients in the CCCI group were attacked by cerebral ischemic(2%),and no one in the normal control group was attacked by cerebral ischemic.Serum MMP-3 levels in the acute cerebral infarction group were significantly higher than CCCI group,and both groups were significantly higher than normal control group (P ＜0.05).The basic serum MMP-3 levels in all patients who were attacked by cerebral ischemic were significantly higher than those who had not been attack by cerebral ischemic during this period of time(P ＜0.05).The serum MMP-3 levels of the unstable plaque group were significantly higher than stable plaque group.And both groups were significantly higher than endometrial coarse group(P ＜0.05).Conclusions Elevated levels of matrix metalloproteinase-3(MMP-3)might have something with the stability of carotid atherosclerotic plaque,and participate the attack and the relapse of acute cerebral infarction.Determination of MMP-3 might be used to predict the attack and relapse of acute cerebral infarction.

OBJECTIVE To choose the marks of screening HBV infective source during surgical operation.METHODS On the basis of the HBV infective threshold value,HBV infective source and HBV susceptibility among the surgeons,to choose the marks of screening HBV infective source during surgical operation.RESULTS Before the immunization of surgeons against HBV,the ratio of HBV infective source between HBsAg positive and HBeAg positive carriers was 19.2% and 83.9%,respectively.That of HBsAg negative and HBeAg negative carriers was 0 and 3.2%.The sensitivity of screening HBV infective source with HBsAg acting as mark was much higher than HBeAg.The specificity was lower than HBeAg.After the immunization of surgeons against HBV,the ratio of HBV infective source between HBsAg positive and HBeAg positive carriers was 0.6% and 3.2%,respectively.None of the negative one was HBV infective source during surgical operation.The sensitivity of screening HBV infective source with HBeAg acting as mark was the same as HBsAg.But the specificity was remarkably higher than HBsAg.CONCLUSIONS Before the immunization of surgeons against HBV,HBsAg acting as the screening mark of HBV infective source during surgical operation is more suitable.After that HBeAg is more suitable.

Objective To apply the prosthesis and orthosis to improve the walking ability of patients after meningomyelocele.Methods A case was reported.Results and Conclusion After wearing prosthesis and orthosis combined with physical therapy for 3 weeks,She could walk 1000 m with only one hand crutch and the speed reached to 36 m/min.