Abstract:Objective To construct and identify a novel single targeted CRAd shuttle vector pshuttle-E1A-E1Bp-E1B55K selectively delated CR2 region of E1A gene,and lay the foundation for the research of gene therapy of cancer. Methods The   E1B55K and E1A-E1Bp genes were amplified by reverse transcriptase polymerase chain reaction (RT-PCR) from human embryonic kidney cells (293T);E1A gene deleted CR2 region was amplified by overlap-PCR.The shuttle vector of pshuttle-E1A-E1Bp-E1B55K was constructed by gene recombinant technique,PCR and enzyme analysis were used to identify the shuttle vector. Results These sequences of amplified E1B55K and E1A-E1Bp deleted CR2 region were consistent with that published on GenBank,indicating that the E1B55K and E1A-E1Bp deleted CR2 region genes were cloned successfully. Furthermore,the recombinant plasmid pshuttle-E1A-E1Bp-E1B55K was identified by endonuclease digestion and PCR,which could be digested into two fragments of XbaⅠ or KpnⅠ，one length of 817 base pairs for XbaⅠ and the length of 1 244 base pairs for KpnⅠ,and the lengthes of  amplified fragments by PCR were 250 base pairs and 1 148 base pairs. These were coincident with the anticipated result. Conclusion A novel recombinant CRAd shuttle vector pshuttle-E1A-E1Bp-E1B55K is constructed successfully which is useful for cancer gene therapy.

Abstract:Objective
To construct the hTERT promoter-driven TRAIL expression vector and explore its inhibitory effects on the proliferation of hepatoma SMMC7721 cells and HL7702 hepatocytes. Methods The recombinant plasmids pshuttle-TRAIL and pshuttle-hTERT-TRAIL were constructed with molecular biological methods. They were confirmed correctly by endonucleases digestion and PCR identification,and transfected into SMMC7721 and HL7702 cells through liposome,meanwhile,the pcDNA3.1+-GFP was transfected to detect the transfection efficacy by fluorescence microscopy. MTT was used to detect the effects of the recombinant plasmids on cell proliferation in transfected cells. Results The recombinant plasmid pshuttle-TRAIL and pshuttle-hTERT-TRAIL were constructed successfully. And the transfection efficacy was highest by fluorescence microscopy when the ratio of plasmid to liposome was 1∶3. As compared with control group, the inhibitory rates of SMMC7721 cells from 16 h after transfection in pshuttle-hTERT-TRAIL group was increased significantly (P<0.001);and as compared with pshuttle-TRAIL group,the inhibitory rates of SMMC7721 cells from 24 h after transfection in pshuttle-hTERT-TRAIL group was increased significantly (P<0.05). However,as compared with control group,the inhibiory rates of HL7702 cells in recombinant plasmids groups did not change significantly. Conclusion hTERT promoter-driven TRAIL could significantly inhibit the hepatoma cell proliferation,especially superior to that in pshuttle-TRAIL,however,no effects on hepatocyte proliferation.

Abstract:Objective
To investigate the effect of isoflurane on the synaptic long-term potentiation (LTP) in the CA1 area of the aged rat hippocampal slices，and elucidate the mechanisms of the effect of isoflurane on cognitive function in the aged rats. Methods Hippocampal slices (400  μm thick) were obtained from the brains of 70 male Sprague-Dawley rats that were decapitated. The slices were incubated in artificial cerebrospinal fluid (ACSF) at room temperature for at least 120 min before use. Thirty-five slices were randomly divided into 5 groups (n=7):control group,isoflurane groups(0.062 5,0.125 0,0.250 0 and 0.500 0 mmol/L). All the slices in each group were perfused with ACSF,isoflurane 0.062 5,0.125 0,0.250 0 or 0.500 0 mmol/L,respectively. The slices in each group were performed to record evoked population spikes (PS) using a extracellular microelectrode recording technique. Another thirty-five slices were randomly divided into 5 groups (n=7):LTP group,isoflurane-LTP groups (0.062 5,0.125 0,0.250 0 and 0.500 0 mmol?L-1). All the slices in each group were perfused with ACSF,isoflurane (0.062 5,0.125 0,0.250 0 and 0.500 0 mmol?L-1),respectively. PSs were recorded for at least 30 min before LTP in each group. For LTP induction,high-frequency stimulation (HFS) conditioning pulses (100 Hz/s) were applied to the Schaffer collateral-commissural pathway of hippocampus using a bipolar stimulating electrode. The changes in PS amplitude after HFS were analyzed in each group. Results  The PS amplitude of the rat hippocampal slices in isoflurane 0.062 5 mmol/Lgroup had no significant difference compared with control group(P>0.05). The PS amplitudes in isoflurane groups (0.125 0,0.250 0 and 0.500 0 mmol/L) were significantly decreased compared with control group (P<0.01). The PS amplitude in LTP group after HFS was significantly increased by (34±11)% compared with that of pre-HFS indicating the successful induction of LTP. Compared with LTP group,the amplitudes of the PS in isoflurane-LTP groups(0.062 5,0.125 0,0.250 0 and 0.500 0 mmol/L) after HFS were decreased significantly (P<0.01). Conclusion The inhibition of LTP induction in hippocampus of the aged rats may contribute to isoflurane-induced deficits in cognitive function of the aged rats.

Abstract:Objective To study the effect of activin A on voltage-gated sodium current(INa) of Neuro-2a cells and mechanism in order to lay a foundation for the application of activin A in nerve system diseases. Methods Neuro-2a cells were divided into IgG control group and anti-ActRⅡA antibody-stained group. Immunohistochemical staining was used to observe whether ActRⅡ A was expressed in Neuro-2a cells. The Neuro-2a cells were randomly divided into normal control group and activin A administration group. Whole cell patch clamp was used to analyze the effect of activin A on INa  of Neuro-2a cells. The expressions of vasoactive intestinal peptide(VIP) and inducible nitric oxide synthase(iNOS) mRNA were examined by RT-PCR in Neuro-2a cells treated with activin A. Results ActRⅡA expression was detectable in normal Neuro-2a cells. Compared with control group,activin A increased INa in Neuro-2a cells significantly (P<0.05).The VIP mRNA expression in activin A group was higher than that in control group (P<0.05),while the expression of iNOS mRNA was lower than  that in control group (P<0.05). Conclusion Activin A can increase INa of Neuro-2a cells and the effect of activin A on regulating the neuron function may be mediated via VIP and iNOS pathways.

Abstract:Objective
To sieve out the fully human,Peroxiredoxin Ⅰ (Prx Ⅰ) -specific single chain variable fragment (scFv) antibodies from the lung adenocarcinoma-related phage antibody library and analysis the biodistribution of the antibodies in vivo. Methods The insert ratio of scFv antibodies library was identified by polymerase chain reaction (PCR) and the products digested by Sfi Ⅰ/ Not Ⅰ were analyzed on 1% agarose gel. Panning against lung adenocarcinoma cell line A549 and Prx Ⅰ were performed three rounds,separately. The positive clones were chosen for soluble expression and analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE),enzyme-linked immunosorbent assay (ELISA) and competition ELISA. The tumor-beared athymic mice were injected via the tail vein with purified 131I-labeled scFv. The biodistribution analysis and radioimmunoimaging were taken to evaluate the specificity and distribution of antibodies in vivo. Results The ratio of recombinant bacteria inserted with scFv was 77% (23/30) and the enzyme digest reaction showed the aim products on 1% agarose gel. From the first to the sixth round of panning,the obtained phages number increased 180 times. The positive reactions to A549 cells were detected in 6 of 10 clones. The positive ratio was 60% (6/10). The human scFv vs  against Prx Ⅰ of lung adenocarcinoma were confirmed by SDS-PAGE and ELISA. Competition ELISA showed significant,concentration-dependent affinity of scFv antibodies to A549 cells. The radiochemical purity of 131I-labeled scFv was (95.6±3.7)% and the specific activity was (2.8±0.2) TBq/g. The radiolocalization indexes (RI) of tumor/serum and tumor/muscle were gradually increased,reaching its peak (4.06±0.13 and 5.17±0.97,respectively) at 48 h postadministration. The radioactivity was aggregated in tumor locations and the tumor imaging was clearly observed by single photon emission computed tomography (SPECT) imaging. The T/NT value at 48 h (3.73±0.20) was significantly higher than those at 12 h (1.26±0.15),24 h (2.18±0.16) and 72 h (2.85±0.18) (all P<0.01). Conclusion The scFv fragments against Prx Ⅰ of lung adenocarcinoma are acquired by screening the phage antibody library successfully. The soluble scFv antibodies have bonding avidity specifically in vivo.

Abstract:Objective To study the effects of cadmium chloride (CdCl2) on endoplasmic reticulum stress by observing the expressions of CHOP and GRP78 mRNA in SMMC-7721 cells. Methods SMMC-7721 cell cultures were exposed to CdCl2 at concentraions of 0,5,10,20,30 and 40  μmol /L for 24 h. Power SYBR green PCR Master Mix and Mx3000P QPCR System were used to measure a threshold cycle (CT) value for each sample. Relative quantitation of gene expression was based on the comparative CT method. Results The CHOP mRNA level tended to increase along with the elevating of CdCl2 dose. The expressions of CHOP mRNA in all experimental groups were higher than that in control group,and the expressions of CHOP mRNA in 10,20 and 40  μmol /L groups were increased significantly (P<0.05). The GRP78 mRNA level was increased along with the elevating of CdCl2 dose. The expressions of GRP78 mRNA in 5,20 and 40  μmol /L groups were significantly higher than that in control group (P<0.05 or P<0.01). Conclusion CdCl2 can induce the expressions of CHOP and GRP78 mRNA in SMMC-7721 cells.

Abstract:Objective
To investigate the effect of local application of simvastatin on the expression of bone morphogenetic protein-2(BMP-2)  in the tooth socket following tooth extraction and discuss its mechanism. Methods Fifty-eight male Wistar rats were divided into experimental group and control group (n=28). PLGA was immediately implanted with or without simvastatin into extraction sockets of the mandibular incisors. Oligonucleotide probe specific to BMP-2 mRNA was designed,and hybridization technique was conducted 5 d,1 week,2 weeks and 4 weeks after implantation.Results There were no positive cells in both experimental group and control group 5 d after implantation. There were deep-dying positive stromal cells between the implanted material and the wall of tooth socket in experimental group,and a few light-dying positive cells could be seen in control group 1 week after implantation. There were many positive cells with clear particles  in the stromal cells between the implanted  material and the wall of tooth socket,and there were also BMP-2 mRNA  positively expressed cells on the surface of newly-formed osteoid in two  groups,and the number of the positive cells in experimental group was higher than that in control group 2 weeks after implantation. There were BMP-2 mRNA  positive cells  in both the experimental group and the control group 4 weeks after implantation. The number of BMP-2  positive cells in experimental group was significantly higher than that in control group 1,2,4 weeks after implantation (P<0.05). Conclusion Topical application of simvastain could promote alveolar bone repair by up-regulating BMP-2 mRNA expression.

Abstract:Objective
To study the effect of thyroid hormone deficiency on malonaldehyde (MDA) and total antioxidative capacity (TAC) and superoxide dismutase (SOD) levels in different developing stages of rat brain and  further explore the mechanism of impaired brain development caused by thyroid hormone deficiency. Methods Perinatal hypothyroidism was induced by administering propylthiouracil ( PTU) solution to the dams by gavage (2 % PTU 1. 5 mL/d) beginning on embryonic day 15. The blood samples of  pups in different developing stages(7,14,21 d) were collected to measure serum FT3 and FT4 levels. And the brain tissues were collected to measure MDA,TAC and SOD levels in brain homogenate. Age matched controls were pups from normal dams without administering propylthiouracil ( PTU) solution. Results Compared with age matched controls,the MDA and TAC and SOD levels were increased in hypothyroid group of 21 d pups（MDA:14.63 μmol/g/±3.44  μmol/g  vs  10.52 μmol/g ±2.32  μmol/g，P<0.05;TAC:0.051 mmol/g±0.006 mmol/g  vs  0.042 mmol/g±0.003 mmol/g，P<0.05;SOD:79.63 μmol/min/g±10.10  μmol/min/g   vs  63.51 μmol/min/g±8.35 μmol/min/g,P<0.05）,but for the 7 d and 14 d pups,ther
e were no differences between two groups (7 d MDA:11.20 μmol/g±2.39  μmol/g　vs  12.29 μmol/g±1.79  μmol/g,P>0.05;14 d MDA:13.07 μmol/g±3.16  μmol/g  vs  12.98 μmol/±2.93  μmol/g,P>0.05;7 d TAC:0.046 mmol/g  ±0.006 mmol/g vs  0.047 mmol /g  ±0.004 mmol/g,P>0.05;14 d TAC:0.042 mmol/g  ±0.007 mmol/g  vs  0.046 mmol/g ±0.007 mmol/g,P>0.05;7 d SOD:66.34 μmol?min-1/g ±11.20  μmol/min/g  vs  65.49 μmol/min/g±7.52  μmol/min/g,P>0.05;14 d SOD:67.53  μmol/min/g±12.01  μmol/min/g  vs  64.57  μmol/min/g±8.28  μmol/min/g/,P>0.05). Conclusion Thyroid hormone deficiency during rat brain development may cause the changes of MDA,TAC and SOD levels and this may be related to brain damage caused by thyroid hormone deficiency.

Abstract:Objective
To construct recombinant plasmid co-expressing siRNA-Stat3 and GRIM-19,and observe its influence in the proliferation ability of prostate cancer cells.  Methods A recombinant plasmid co-expressing siRNA-Stat3 and GRIM-19 was constructed by using gene cloning. The prostate cancer cell lines PC-3M were transfected with the co-expression plasmid. The gene expression of Stat3 and GRIM19 was determined by semi-quantitative RT-PCR assay. The influence of recombinant plasmid in the proliferation ability of prostate cancer cells was determined by MTT. Results The siRNA-Stat3 and GRIM-19 co-expression plasmid was successfully constructed. Compared with control group,the expression of Stat3 mRNA was significantly decreased,and the expression of  GRIM-19 mRNA was significantly increased（P<0.01）,the proliferation rate of prostate cancer cells was significantly decreased（P<0.05）in experimental group. Conclusion The siRNA-Stat3 and GRIM-19 co-expression plasmid  is successfully constructed,and  it has significant inhibition on the growth of prostate cancer cells.

Abstract:Objective To study the effect of trantinterol,a potent and highly selective  β2 receptor  agonist,on the cytochrome P450 enzymes (CYP450) mRNA expression and provide a theoretical basis for clinical drug combination.Methods  The cell proliferation of HepG2 cells treated with  2.4,12,60,300,1 200 and 4 800 ng/L  trantinterol was analyzed by MTT.The mRNA expression levels of CYP1A1,CYP2E1 and CYP3A5 in HepG2 cells were examined using the real-time quantitative reverse-transcriptase polymerase chain reaction in human hepatoma HepG2 cells.Results Different concentrations of trantinterol didn’[KG-*3]t inhibit the proliferation of HepG2 cells (P>0.05).Compared with control group,the mRNA expressions of CYP2E1 and CYP3A5 in trantinterol groups had no change,while the mRNA expression of CYP1A1 was inhibited (P<0.01).Conclusion Exposure of HepG2 cells to trantinterol has no effect on the mRNA expressions of CYP2E1 and CYP3A5 but inhibit the mRNA expression of CYP1A1.

Abstract:Objective  To observe the effects of autoimmune regulator(AIRE) on CD4+CD25+Treg cells,and explore the functions and mechanisms of   AIRE in peripheral tolerance.Methods RAW264.7 cells were transfected with pEGFPC3AIRE plasmid via lipofectin.Flow cytometry was used to determine the amount of CD4+CD25+Treg cells.  Then semi-quantitative RT-PCR was employed to detect the expressions of Foxp3 and TGF-β1  mRNA.The level of TGF-β1   was detected with ELISA kit. Results AIRE transfected RAW264.7 cells could directly or indirectly induce the generation of CD4+CD25+Treg cells and the increased expression of Foxp3 mRNA.Conclusion The AIRE expressed in the peripheral antigen presenting cells(APCs) could directly and/or indirectly induce the generation and functions of CD4+CD25+Treg cells,and probably plays a crucial role in the peripheral immune tolerance.

Abstract:Objective
To study the protective effect of zinc sulfate on liver injury of diabetic rats induced by STZ and explore its mechanism. Methods Fifty  male Wistar rats were randomly divided into five groups: normal diet (NC) group, high fat diet (FC) group, diabetes (DM) group, Zn before the intervention of diabetes (ZnDM) group, Zn after the intervention of diabetes (DMZn) group. The rats in Zn groups were fed with Zn ions (Zn2+) 15　mg?kg-1?d-1 by giving ZnSO4 solution daily by gavage, the others were fed with distilled water by gavage. Eight weeks after STZ injection, the serum was collected to detect the function of liver, AST and ALT, and liver was taken for the measurement of liver index, SOD, MDA, and GSH-Px. Results  Compared with DM group, Zn intervention could significantly reduce the liver index (P<0.05); the levels of AST and ALT in serum were decreased mildl (P>0.05); the level of MDA in liver tissues was markedly decreased and the levels of SOD and GSH-Px in liver tissue were significantly improved (P<0.05)； the levels of MDA and SOD in liver tissues had significant differences between the two groups of Zn intervention (P<0.05). Conclusion Exogenous zinc sulfate can decrease the liver index of diabetic rats, and make the liver function better. The mechanism of the protection of Zn may be related to the strengthening of anti-oxidation.  

Abstract:Objective
To investigate the changes of the expression of corticosteroids metabolic enzymes  11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) and glucocorticoid receptor (GR) in skeletal muscle in type 2 diabetic rat  model and significance. Methods The Wistar rats were devided into control and type 2 diabetic model groups. The models were built  with a combination of high fat diet (HFD) and multiple low dose of STZ (30 mg/kg, twice) injection. 8 weeks later, the blood of rats were colleted by cutting their tail，the blood glucose was tested with the method of oxidizing enzyme. According to the serologic test and the relevant index of insulin, the damage of islet and the secretion of insulin were determined. The protein expressions of corticosteroids metabolic enzymes 11β-HSD1 and GR in skeletal muscle were examined by Western blotting analysis. Results  Compared with control group, the insulin secretion(IS), insulin sensitivity index(ISI) and secretion index of HOMA β cell (HβCI) were decreased significantly(P<0.05), while the insulin resistance index (IRI) was increased significantly(P<0.05). The protein expressions of 11β-HSD1 and GR in model group were higher than those in control group(P<0.05). Conclusion The type 2 diabetic  model built  with a combination of HFD and multiple low dose of STZ  injection has the characteristics of high blood glucose, damage of IS function and insulin resistance. In the type 2 diabeic models the protein expressions of 11β-HSD1 and GR are significantly increased.

Abstract:Objective
To investigate the expression and role of NF-κB in  rat's  with myocardial ischemic fibrosis,and  illuminate the relations between  NF-κB and  occurrence and development of myocardial ischemic fibrosis. Mthods 20 male Wistar rats were  randomly divided into  control group，model group(12 h,72 h and 1 week)(n=5)， Except control group,all the rats in the other groups were treated with isoproterenol hydrochloride(2 mg?kg-1).Conventional  staining,immunohistochemical staining and Masson staining were used to analyze expression of NF-κB protein in heart tissues. Results The results of HE staining showed the myocardium tissue had been a good order,the clear nuclear arms,no cell swollen. The mycardium presented many punctual necrosis at 12 h. The myocardium appeared  clear boundary  and multi-radiations  necrosis stove at 1 week. The results of Masson staining showed the myocardium had only a small number of collagen in normal control group,the expression of collagen was  obviously increased at 12 h(P<0.01) and reached the arrived  peak at 1 week. The results of NF-κB protein expression showed there was positive matter of buff or yellow. The NF-κB expression was  increased obviously at 12 h and reached the  peak at 1 week(P<0.05). Conclusion Along with,The NF-κB expression is increased obviously which indicats that NF-κB is involved in the occurrence and development of myocardial ischenic frbrosis in rats.with the prolongation of Iso injection time.

Abstract:Objective
To express and purify the fusion protein of rhaFGF-tat with biological activity,and provide a foundation for development of an genetically engineered drug. Methods The aFGF-tat gene fragment through digesting the pMD18-T-aFGF-tat was obtained,then the recombinant plasmid was transformed into E.coil BL21 (DE3) and induced by IPTG. The protein was purified by CM-Sepharose FF cation exchange and heparin-Sepharose FF affinity. The activity of aFGF-tat was detected with NIH 3T3 proliferation assay. Results Acquired gene fragments of aFGF-tat  identified by digestion and DNA sequencing were coincident with the human aFGF-tat reported in GenBank. The recombinant vector of pET3c-aFGF-tat was constructed successfully. SDS-PAGE result proved that aFGF-tat fusion protein with a relative molecular mass of about 19 102 was expressed. The purity of fusion protein was more than 95%,Western blotting results showed that the expressed products had specific reaction with anti-human aFGF polyclonal antibody and was able to promote the proliferation of NIH 3T3 cells. Conclusion　aFGF-tat fusion gene is expressed in E.coil and purified successfully,and the aFGF-tat fusion protein can promote the proliferation of cells.

Abstract:Objective
To study the mechanism of fusion protein of hsp65-MOMP-T-epitopes(H-ctm1) against C.trachomatis(Ct) genital tract infections that will underlay further research for T-epitopes Ct vaccine. Methods Models for Ct genital tract infections of mice were established.In the experiment of delayed type hypersensitivity(DTH),14 C57BL/6 female mice were divided into two groups(H-ctm1 immunized mice,naive mice,7 mice every group). The thickness of bilateral hind footpad of every mouse was measured,then,the left hind footpad was injected with SPG,and the right hind footpad with heat-killed chlamydia（HK-EBs）.The thickness of hind footpad was remeasured at 72 h following injection. In  the experiment of flow cytometric quantitation,21 C57BL/6 female mice were divided into three groups(H-ctm1 immunized mice,HK-EBs immunized mice,PBS-immunized mice,7 mice every group).After immunization,these mice were infected by Ct. 6 d after infection,the CD4+ and CD8+ T cells were dectected with flow cytometry(FACS). Results For the experiment of DTH,in the group of naive mice,there was no significant difference in thickness between pre-injection and post-injection whatever right or left hind footpad（P>0.05）,also between right and left hind footpad whatever pre-injection and post-injection（P>0.05）.In the group
 of H-ctm1 immunized mice,there was no significant difference in thickness between right and left hind footpad before injection（P>0.05）,however,the difference was  significant  after injection（P<0.05）.For left hind footpad,the difference was not significant   between before and  after injection（P>0.05）;for right hind footpad,the difference was  significant  between before and after injection（P<0.05）.[JP3]In the FACS experiment,the mean ratio of CD4+ T cells in H-ctm1 group(18.9%±[JP]2.2%) was higher than that in HK-EB group(11.3%±3.1%,P<0.05)) and control group(0.05%±0.07%,P<0.01). And the mean ratio of CD8+ cells in H-ctm1 g
roup(5.7%±1.9%) was higher than that in HK-EB group(1.5%±0.5%,P<0.01) and control group(0.02%±0.01%,P<0.01). Conclusion DTH play a role in the process of immune response to H-ctm1 immunized mice.HSP65 fusion protein can stimulate the dendritic cells to up-regulate the levels of MHC（class Ⅰ and class　Ⅱ）and improve  MOMP-T-epitopes immunogenicity.

Abstract:Objective
To study the anti-fatigue effect of the water extract of Acanthopanax senticosus fruits and its mechanism，and provide experimental basis for exploration and utilization Acanthopanax senticosus resources. Methods Acanthopanax senticosus fruits were extracted by ultrasonic method and their aqueous extract was prepared.The mouse models of anti-fatigue were randomly divided into negative control,positive control and different doses of aqneous extract from Acanthopanax senticosns fruits groups.The anti-fatigue effect of aqueous extract from Acanthopanax senticosus fruits was observed through determining the exhaustive swimming time and the contents of hepatic glycogen，muscle glycogen，superoxide dismutase (SOD),lactate dehydrogenase（LDH）,blood lactic acid(LAC) as well as blood urea nitrogen(BUN) with stamina swimming mice as animal models of anti-fatigue. Results Compared with negative control and positive control groups,the loaded swimming time of mice was prolonged in different doses groups of aqueous extract from Acanthopanax senticosus fruits（P<0.01)；and the serum LDH activity was increased and the contents of LAC and BUN were decreased after exercise(P<0.01). The levels of hepatic glycogen and muscle glycogen of postexercise mice were improved;and the more sugar for aerobic athletics was increased;the SOD activity of exercising mice was significantly improved(P<0.05).Conclusion Acanthopanax senticosus fruits have anti-fatigue effects.

Abstract:Objective
To investigate the effects of ionizing radiation on the expression of γ-H2AX in Jurkat cell lines. Methods The changes of the expressions of γ-H2AX in Jurkat cell lines after X-irradiation with different doses(0.5,1.0,2.0,4.0,and 6.0 Gy) and after irradiation with 2.0 Gy X-rays for different time(0,0.5,1.0,4.0,8.0,16.0,and 24 h) were analyzed by flow cytometry (FCM),and the apoptotic rate of Jurkat cell lines was detected during dose-effect experiment. Results In time-effect experiment,the results showed that the expression of γ-H2AX reached to the peak within 1 h,followed by a time-dependent decrease,kept expressing at 16.0 h,and had no significant difference with control at 24.0 h. In dose-effect experiment,it was demonstrated that the expressions of γ-H2AX showed a dose-dependent increase in the range of 0.5-4.0 Gy at 1.0 h after exposed to X-rays,and dropped at the does of 6.0 Gy;at 8.0 h after exposure to those doses above,the express levels of γ-H2AX enhanced linearly with the dose increased in the range of 0-4.0 Gy,showing a clear dose-dependent manner(P<0.01). Conclusion The γ-H2AX expression could be increased by X-irradiation in time-dependent and dose-dependent manners.

Abstract:Objective
To investigate the protective effects of total flavonoids of Bidens bipinnata L. (TFB) on acute and chronic liver injury in mice induced by carbon tetrachloride and  mechanisms. Methods Sixty Wistar rats were randomly divided into control group,model group,Hu-Gan-Pian(4 g?kg-1) group and TFB (120，60，30 mg?kg-1) groups. The acute and chronic liver injury model in mice were induced by different dosages and administration times of carbon tetrachloride respectively to study the effects of TFB on liver index,hepatic morphological changes and detect the contents of SOD,MDA,ALT and AST in serum. Results  Compared with model group,the degree of hepatic edema in TFB（120，60，30 mg?kg-1）and Hu-Gan-Pian(4 g?kg-1) groups were decreased（P<0.05 or P<0.01）,the contents of AST,ALT and MDA in serum were decreased（P<0.05 or P<0.01）,and the contents of SOD were increased（P<0.05 or P<0.01）. TFB and Hu-Gan-Pian could ameliorate the pathological lesion. Conclusion TFB has protective effects on carbon tetrachloride-induced acute and chronic liver injury in mice,which may be related to inhibition of lipid oxidation.

Abstract:Objective To study the effects of purified product of trihydroxybenzoic acid (TBA) from water caltrop and its compound at different concentrations on cell cycle progression and apoptosis of Jurkat cells in vitro and explore their possible mechanism of inhibiting tumor growth.  Methods The Jurkat cells at logarithmic growth phase were treated with purified product of TBA and its compound in vitro. Each TBA was divided into 5 groups (0,3.125,6.250,12.500 and 25.000 mg/L). The changes of cell cycle progression and apoptotic rate of Jurkat cells 30 h after treated with TBA were measured with flow cytometry.  Results The apoptotic rates of the Jurkat cells treated with purified product TBA with doses of 3.125 and 25.000 mg/L were increased significantly as compared with those in  control group(P<0.05 or P<0.01);the apoptotic rates of Jurkat cells treated with TAB compound with different doses were increased with the increase of doses (P<0.05 or P<0.01). The percentages of G1 phase cells treated with purified product TBA with different doses were increased significantly with the increase of doses as compared with those in  control group(P<0.01),the percentages of S phase cells were decreased significantly in 6.250 and  25.000 mg/L groups（P<0.05 or P<0.01），and the percentages of G2 phase cells did not change significantly. The percentages of G1 phase cells treated with TAB compound with 6.250 and  25.000 mg/Lwere significantly higher than those in  control group(P<0.05 or P<0.01),the percentages of S phase cells were decreased significantly (P<0.05),and the percentages of G2 phase cells did not change significantly. Conclusion Two different TBA could all induce Jurkat cell apoptosis and G1 phase arrest. Their inhibitory effects on tumor growth could be due to apoptosis caused by G1 arrest.

Abstract:Objective To study the inhibitory effects of melatonin(MLT),cisplatin(DDP),MLT combined with DDP on human ovarian cancer cell line SKOV3,and provide the theoretical basis for the study on MLT as an ancillary drug of ovarian cancer. Methods The ovarian cancer cell SKOV3 at logarithmic growth phase were divided into control group,MLT groups(0.5,1.0 and 2.5 mmol/L),DDP groups(25,50 mg/L) and MLT combined with DDP groups(MLT 0.5 mmol/L combined with DDP 25 mg/L；MLT 0.5 mmol/L combined with DDP 50 mg/L；MLT 1.0 mmol/L combined with DDP 25 mg/L；MLT 1.0 mmol/L combined with DDP 50 mg/L；MLT 2.5 mmol/L combined with DDP 25 mg/L；MLT 2.5 mmol/L combined with DDP 50 mg/L). The apoptotic rate and proliferation inhibitory rate of ovarian cells were detected with microscopic observation and MTT assay,and the expression of VEGF in ovarian cancer cell SKOV3 was determined. Results Observing with microscope,in the cells treated with drugs the adherence ability was reduced with refraction,shrink,circular,karyopycnosis,nuclear fragmentation and the cell chromatin agglutination. The apoptotic cells of solo drug groups were more than those in combined drug groups. MTT assay indicated that the MLT groups(0.5,1.0,2.5 mmol/L) and MLT combined with DDP(25,50 mg/L) groups all had inhibitory effects on the cell growth,and the inhibitory rate was increased with the concentrations of drugs. Two drugs had synergetic effect,CDI＜1. Immunocytochemical staining results showed that the expressions of VEGF in solo drug groups and combined groups were lower than that in control groups,the expression of VEGF in combined groups was decreased significantly(P<0.05). Conclusion  MLT can restrain the growth of ovarian cancer cells. MLT combined with DDP has synergetic effect on down-regulation of the expression of VEGF. Combined DDP with debita spissitudine MLT can achieve good curative effect.

Abstract:Objective  To detect the different proteins in ovarian carcinoma tissues and explore the relationship between the proteins and the carcinogenesis and development of ovarian carcinoma. Methods 19 cases of serous carcinoma tissues and 12 cases of normal ovarian tissues were collected, the proteins from tissues were extracted. With protein pathway array technique,37 kinds of phosphorylative antibodies and 73 kinds of  non-phosphorylative antibodies, the different proteins in ovarian carcinoma tissues and normal ovarian tissues were detected. The results were analyzed by QUANTITY One software and statistically calculated. Results There were 52 proteins expressed (47%) and 23 proteins (21%) were diferentially expressed with statistical significance (P<0.05), including the over expression proteins: p-Stat3, p-PDK1, p-cdc2, p-p38, p-AKT, XIAP, PCNA, cdc2p34, p53, Cdk2, Mesothelin, E-cadherin and down-regulated proteins: ERK, Cdc25C, Wee 1, Akt, N-cadherin, α-tubulin, Vimentin, cdc42, Bax, PTEN (P<0.05). Compared with normal ovarian tissues and normal ovarian cell line,p-Stat3, p-PKD1 and p-AKT were over expressed in ovarian carcinoma tissues and cell lines (P<0.05). Conclusion Stat3, PKD1 and AKT play important roles in carcinogenesis of ovarian carcinoma

Abstract:Objective To study the changes of interleukin 8 (IL-8) and interleukin 10 (IL-10) in hyperoxia-induced lung injury and significances. Methods  The model of rat with hyperoxia-induced lung injury was established by being inhaled with high concentration of oxygen (>95%).The lung tissues were harvested 6 h and 1,3,6,10,14 d  after oxygen treatment, the histopathological changes of lung tissues  were observed by HE staining and Double-antigen sandwich ELISA was used to detect the IL-8 and IL-10 levels in the lung tissue homogenate. Results  The infiltrative inflammatory cells appeared in alveolus first,and then alveolus was enlarged ,then alveolar septum became thinner and eventually the number of alveolus was decreased as the time of exposure to oxygen prolonged. The concentrations of IL-8 in lung tissue homogenate in high  oxygen group were significantly higher than those in control group, the level reached the peak at  day 3 after high oxygen exposure and the statistical difference lasted until the 14th day. The concentrations of IL-10 in lung tissue homogenate were significantly higher after high oxygen exposure 6 h and also had statistical difference on day 1, but restored normal level at day 3. Conclusion The concentration of inflammatory factor IL-8 is increased fast and lasts for long time while anti-inflammatory factor IL-10 is increased for short time in hyperoxia-induced lung injury.

Abstract:Objective To study the effects of methylmercury chloride (MMC) on protein kinase C ( PKC) activity in rat developing hippocampus. Methods The animals in experimental groups were fed with standard rat chow including  0.75 mg/kg MMC,1.5 mg/kgMMC and 3.0 mg?kg-1 MMC respectively for 90 d before gestation to 30 d post parturition. Hippocampus of pups from each group on postnatal days ( PND) 3,7,17,21 and 30 were dissected. All the samples were separated into cytosol and membrane subcellular fractions and assayed for PKC activity by the improved Takai’s method. Results The membrane and cytosolic PKC activities of pup hippocampus in experimental groups were significantly higher than those in control group.The PKC activities of rats in 3.0 mg/kg MMC group and PND 21,30 in 1.5 mg?kg-1 MMC group were significantly increased compared with control group (P<0.05).Conclusion The damage of the rat developing hippocampus by chronic exposure to MMC is possibly correlative with the enhanced effect of MMC on PKC activity.

Abstract:Objective To  establish the animal model of arteriogenic erectile dysfunction(ED) and explore the pathogenesis of ED. Methods Thirty male Wistar rats were divided into two groups:experimental group (n=15),treated with ligating rat bilateral internal arteries;control group (n=15),treated with sham operation only. 4 weeks later,the erectile function of the rats was evaluated by injecting with apomorphine (APO) and  electrostimulation of cavernous nerves and recording of the　intracavernous pressure (ICP). The ultrastructures of the smooth muscle cells of cavernous of penis were observed with transmission electron microscope;the NOS level in the penis tissue was examined by NADPH diaphorase staining. Results In APO experiment,the penile erection times in experimental  group were decreased significantly compared with control group(P<0.01);the number of yawns was showed no obvious difference between  two groups（P>0.05）. In ICP experiment,the mean amplitude of ICP in experimental group was decreased significantly compared with control group (P<0.01). Electron microscopy showed that in the smooth muscle cells in experimental group,there were a significantly less number of mitochondria in the cytoplasm,less number of caveolae with fingerlike processes in the plasma membrane,increased number of vacuoles in the cytoplasm compared with control group. The NOS staining in experimental group was less intense compared with control group. Conclusion It is easy,practicable and reliable to establish the arteriogenic ED rat animal model by ligating rat bilateral internal arteries. Both the changes of ultrastructures and the low NOS level in penis tissue play roles in the occurrence of ED.

Abstract:Objective To study the influence of ginsenoside Rg1 on injury of white matter and illuminate the protective effect of ginsenoside Rg1 and its mechanism after cerebral ischemia. Methods Male ICR mice were pre-treated with ginsenoside Rg1 (50 mg?kg-1) by intragastric (i.g.) injection once a day for 7 days. One hour after the last injection bilateral common carotid arteries were occluded for 60 min. The mice were divided into four groups:sham group(Sal-Sham),drug control group (Gs-Sham),ischemia group (Sal-Isc) and drug-protective group (Gs -Isc).The injury of white matter in dentate gyrus of hippocampus after global cerebral ischemia was detected with immunohistochemical  staining.The MDA protein expression after           cerebral ischemia was determined with Western blotting method.Results Compared with sham group,the MBP positive cells  in ischemia group were decreased（P<0.05）; compared with ischemia group,the MBP positive cells  in drug-protective group were increased（P<0.05）;compared with sham group,the MDA protein  in ischemia group was increased（P<0.05）;compared with ischemia group,the MDA protein  in drug-protective group was decreased（P<0.05）.Conclusion Ginsenoside Rg1 can attenuate the injury of white matter after cerebral ischemia and its mechanism might be associated with decreasing the MDA protein expression.

Abstract:Objective To observe the immunoregulatory effects of Single herb decoction of  Curcumae on acute liver injury indued by CCl4 in mice,and provide experimental evidence for the development and utilization of single herb decoction of Curcumae in prevention and treatment of acute and chronic liver injury. Methods  Forty-eight ICR mice were randomly divided into 6 groups:normal group,model group,high,middle and low doses of Curcumae groups,positive drug group(diammonium glycyrrhizinate). The mice in positive drug group were injected intraperitoneal with diammonium glycyrrhizinate (22.5 mg/kg)，once a day;the mice in treatment groups were orally administered with Curcumae （12,6,3 g/kg）,once a day;the mice in normal and model groups were administered with the same volume of saline,once a day;for 7 d,1 h after administration at the seventh day,the mice in model and treatment groups were  intraperitoneally  injected with   0.2% CCl4 (10 mL/kg); the mice in normal group with the same volume of saline. All the mice were sacrificed 24 h after injection.The liver index was calculated； the  pathological changes were examined by HE staining method . The mRNA expressions of IL-1β and TNF-α were analyzed with RT-PCR. The protein expressions of IL-18 was detected with immunohistochemistry staining .  Results Compared with model group,the liver indexes in treatment groups were obviously down-regulated（P<0.05）.In model group,there were a lot of inflammatory cells infiltration and necrotic hepatocytes in mice,but the lesions became much slighter in treatment groups. Compared with model group,the mRNA expressions of IL-1β and TNF-α and the protein expression of IL-18 in liver of mice were significantly decreased  in treatment groups (P<0.05). Conclusion The Single herb decoction of  Curcumae has the protective effects on CCl4-induced acute liver injury by inhibiting the　expression of inflammatory factors IL-1β,IL-18,and TNF-α and regulating the immune function of liver in mice.

Abstract:Objective To investigate the genetic association between the polymorphisms of PLA2G4B gene and schizophrenia in the North Han Chinese population. Methods The method of PCR-based ligase detection reaction (PCR-LDR) was applied to detect the genotype single nucleotide polymorphisms of rs3816533 in PLA2G4B gene among 201 pedigrees consisting of fathers,mothers and affected offsprings with schizophrenia. Haplotype relative risk (HRR) test and transmission disequilibrium test (TDT) were conducted to analyze the genotyping data. Results The genotypic frequency of rs3816533 of PLA2G4B gene did not deviate from Hardy-Weinberg equilibrium in both case and control groups. HRR analysis did not show allelic association with the PLA2G4B gene (χ2 = 0.840,P>0.05);TDT analysis did not show preferential transmission of the two alleles(χ2 = 0.818,P>0.05).  Conclusion  There is no genetic association between the polymorphism of rs3816533 site on  PLA2G4B gene  and schizophrenia in Han population from the north of China.

Abstract:Objective To identify the locus and explore the pathogenesis of follicular occlusion triad by the linkage analysis in a four-generation follicular occlusion triad family. Methods The peripheral blood samples from all of members in this family were collected and genomic DNA was extracted,all of exons and intron-exon boundaries of KRT5 gene were sequenced. Two-point linkage analysis was performed with 12 microsatellite markers at chromosome 1p21.1-1q25.3 using linkage program (5.2 Version). Results No causative mutations were identified in the KRT5 gene and the cumulative maximum two-point LOD score  were obtained below －2 at 1p21.1-1q25.3 (at recombination fraction=0.00). Conclusion The previously reported locus at 1p21.1-1q25.3 (61.8 cM) for follicular occlusion triad coexisted with Dowling-Degos disease is excluded in this family. This study indicates that follicular occlusion triad is likely a genetically heterogeneous disorder.

Abstract:Objective To investigate the prevalence of arterial vascular calcification (VC) in uremic patients undergoing diabetic nephropathy (DN) and  explore its risk factors.   Methods Demographic and clinical data were collected in the patients latest diagnosed as uremia and not received dialysis treatment. The uremic patients were divided into DN group and non-diabetic nephropathy (NDN)group. And 20 healthy subjects  were chosen as control group from the Physical Examination Room. The patients’ sex and ages were similar in the 3 groups.VC was semi-quantitatively evaluated by plain radiographic films from abdomen，pelvis and hands．The clinical and laboratorial parameters related to VC  were detected and analyzed.  Results ①The present study included 20（51.28%） DN uremic patients and 19(48.72%) NDN uremic patients．The average diabetic duration in the DN patients was (8.11±7.39) years. ②Among the 39 uremic patients，VC was found on radiographic films in 29 cases (74.36％)，including 23 cases (58.97%) with the con-score 1-3 and 6 cases (15.38%)with 3-6 scores. And VC was not detected in control group.Bone density analysis showed that osteopenia occupied 14 cases (35.90%) in all and the T-score was －0.81±0.87. ③Linear correlation analysis revealed that VC was correlated with serum calcium and phosphate（r=0.026,P<0.05；r=－0.639,P<0.05）,and uncorrelated with bone density（P>0.05）. VC score was significantly correlated with the diabetic duration（r=0.790,P<0.001）. Logistic regression revealed that the diabetes duration was the independent risk factors （P<0.05）for VC．The reflections of serum calcium and phosphate were rejected. ④The prevalence of VC in DN gourp (95.0%) was higher than that  in NDN group（42.1%,P<0.05）．And the VC score in DN group （3.18±1.77）was higher than that in NDN group （1.56±0.97,P<0.05).Conclusion There is a higher VC prevalence rate and more VC severity in DN uremic patients than in NDN patients. Diabetes duration is an independent risk factor for VC．Preventing from the high serum glucose maybe plays an effective role in controlling the VC  in diabetic nephropathy patients in uremic stage.

Abstract:Objective To study the pathologic and ultrastructural features of fibrillary glomerulopathy (FGP),and provide basis for diagnosis and differential diagnosis of FGP. Methods Two renopuncture biopsy tissues of FGP（one case combined immunotactoid glomerulopathy) were studied by routine histopathological(morphologically and ultrastructurally) and were observed by light microscope with staining(HE,PAS,PAM-Masson)and immunohistochemical staining(IgG,IgM,IgA,C3,C4,C1q)and  by electron microscope with Lead citrate-Uranyl acetate staining. Results Morphologically,membranous nephropathy with immune reactants were showed in mesangium and subepithelial areas of glomerular basement membrane(GBM) in membranous glomerulopathy(MG) and membranoproliferative glomerulonephritis(MPGN) with mesangial nodular sclerosis and GBM’s double-track,which immune reactants eposited showed in subendothelial areas of GBM in MPGN by PAM-Masson staining. Renal mesangium of MG and MPGN associated with deposition of material that was positive for IgG，IgM，IgA，C3，capillary tuft of MG was positive for IgG，C3 and capillary tuft of MPGN was positive for IgM，C3，C1q by immunohistochemical staining,but they were not stained by Congo red. Ultrastructurally,the mesangial,subepithelial and/or subendothelial areas of MG and MPGN were expanded because of the electron-dense deposits,which were represented by randomly oriented nonbranching fibrils with a diameter of about 15-25 nm. In addition,microtubule deposits with a diameter of about 30-50 nm were also found in MPGN.Conclusion The diagnosis of FGP is always based on biopsy of the kidney which revealed the characteristic of fibrils composed of immune reactants under electron microscope.

Abstract:Objective To study the influence of incline implant supported fixed bridge at the local area of the maxillary molar on the implant-bone-interface stress distribution and illustrate the correlation between every influence factors about incline implant supported fixed bridge and late success rate of implant denture. Methods Three-dimensional finite element model for the insufficient ertical bone mass at the area of the maxillary molar was established biomechanically in this study by spiral CT technique and finite element software technique. Different experimental conditions were designed which were about different implant lengthes (7,9 and 11 mm,diameter of implants was 5.5 mm and oblique angle was 10°),implant diameters（3.5,4.5 and 5.5 mm,length of implants was 7 mm and oblique angle was 10°） and oblique angles of implant (5,10 and 15°,diameter of implants was 5.5 mm,length of implants was 7 mm). The implant-bone-interface stress distribution was analyzed by 3-D finite element method under the action of vertical and decentralized load. Results  When implant length changed from 7 to 11 mm,the stress of necks and root tips of implants was big,and the one on root tips was bigger than the necks at the same time. The stress on root tips was decreased as the length of the implant was increased and was closed to the necks. The feature on stress distribution of different diameters was that the stress of necks and root tips of implants was big with 3.5 and 4.5 mm in diameter,and the stress on each part of the implant-bone-interface was similar with 5.5 mm in diameter. The feature of stress distribution on each part of the implant-bone-interface was very close in resemblance with different oblique angles, the stress of necks and root tips of implants was big. The stress of root tips of implants was increased as the angle was increased. Load distribution also affected stress distribution on the interface. The peak stress under a concentrated load was bigger than the one under decentralized load. Conclusion When the vertical bone mass at the local area of the maxillary molar is relative insufficient,the implant-bone interface stress distribution tends towards uniformity in selecting longer and wider-diameter implant. The  complicated maxillary anatomical structure could be considered adequately to prevent stress concentration to occur in partial thick bone.

Abstract:Objective To investigate the significance of immunohistochemical method in differential diagnosis and definiting the origin of esophageal spindle cell tumors.Methods Eighteen cases of esophageal stromal turmors were studied by HE staning and immunohistochemistry to detect the expressions of CD117, CD34, SMA, Desmin and  S-100.Results In 18 cases of esophageal spindle cell tumors,the CD34, S-100, Desmin positive/negative expression rates were 4/14,0/18 and 12/6 , respectively.6 cases of esophageal spindle cell tumors were positive for CD117,among them  2 cases were local positive in membrane and 4 cases were positive in cytoplasm of spindle cells. 12 cases of esophageal spindle cell tumors were negative for CD117. 17 cases tumor cells were varying degrees diffusely and positive expression for SMA. Among 18 cases of esophageal spindle cell tumors, 12 cases were leiomyoma  and  6 cases were gastrointestinal stromal tumors. Conclusion Immunohistochemistry could accurately determine the biological behavior and prognosis of esophageal spindle cell tumors, it could provide direction for reasonable choice of molecular target therapy after operation.

Abstract:Objective To investigate the expressions of matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9(MMP-9) in endometrial adenocarcinoma tissues and clinical significances and explore the correlation between MMP-2,MMP-9 and development and biological behavior of endometrial adenocarcinoma. Methods 102 cases of uterine samples with complete data were selected. 60 cases of the endometrial adenocarcinoma were divided into three groups by clinical stage:stage Ⅰ(26 cases),stage Ⅱ(19 cases),stage Ⅲ(15 cases);three groups by histological level:level 1(29 cases),level 2(19 cases),level 3(12 cases); three groups by myometrial invasion:non-myometrial invasion(8 cases), myometrial invasion<1/2 (11 cases),myometrial invasion>1/2 (41 cases);and  two groups by lymph node metastasis:non-lymph node metastasis(54 cases) and lymph node metastasis(6 cases). S-P immunohistochemistry method was used to detect the expressions of  MMP-2 and MMP-9. Results The positive rates of MMP-2 and MMP-9 in endometrial adenocarcinoma tissues (76.7% and 80.0%) were significantly higher than those in normal uterine tissues (13.60%) and hyperplasia endometrial tissues (20.0%) (P<0.05). The strong positive rates of MMP-2 and MMP-9 in endometrial adenocarcinoma tissues at stage Ⅲ and histological level 3 were higher than those at  stage Ⅰ and histological level 1 (P<0.05). The strong positive rates of MMP-2 and MMP-9 in endometrial adenocarcinoma tissues with myometrial invasion and lymph node metastasis were obviously higher than those with non-myometrial invasion and non-lymph node metastasis (P<0.05). There are significant differences between them according to the statistics analysis (P<0.05). Conclusion The positive expression rates of MMP-2 and MMP-9 in endometrioid adenocarcinoma tissues are high. The strong positive expression rates of MMP-2 and MMP-9 are related to tumor differentiation grade，clinical stage，myometrial invasion and lymph node mestastasis.

Abstract:Objective To observe the change of  CD4+ T Lymphocytes in patients with H1N1 at early stage and figure out its clinical significances on the progress and therapeutic selection of H1N1. Methods The absolute counts of T lymphocyte subset from the peripheral blood samples of 48 H1N1 patients in first ten days’ duration were detected by flow cytometry,and  the serial chest CT examinations were performed. Results In all 48 clinical cases,28 cases were in normal range of CD4+ lymphocyte absolute count,whose pulmonary lesions were limited and illness condition stayed in the stability,they  didn’〖KG-*3〗t need steroid. In the other 20 cases with low level of CD4+,4 cases’ illness presented the progressive development and needed to be treated with steroid and 16 cases with lightly decreased CD4+ level which had a stable condition without treatment  with steroid. The result of  Pearson correlation analysis showed that there were negative correlations between absolute count of CD4+ cells and pulmonary lesions (r=－0.299，P<0.05) and steroid treatment (r=－0.383，P<0.05). Conclusion The lower peripheral blood CD4+ cell absolute count of H1N1 patients at early stage indicates the worse condition of pulmonary lesions. The patients with remarkable decrease of CD4+ lymphocytes are in need of treatment with steroid.

Abstract:Objective To investigate the correlation and the clinical significance between the overall classification on color Doppler ultrasound and the clinical stages of atherosclerosis obliterans(ASO)，and evaluate the extent of arterial lesions comprehensively. Methods 125 patients                         of ASO，who were divided into three groups of mild，moderate and severe with Color Doppler ultrasound according to differences of occlusion，quantity，degree of stenosis and collateral number，were analyzed with clinical stages，then  their associations were studied with Spearman rank analysis. Results The clinical manifestations of ASO patients who were divided into three groups of mild，moderate and severe according to overall classification on color Doppler ultrasound were respectively gradually serious，which had  positive correlations with the stages of Ⅰ，Ⅱand Ⅲ according to clinical stages. Spearman rank analysis showed that the correlation coefficients (rs) was 0.797 2 between two groups(P<0.01),there was good consistency between the overall classification on color Doppler ultrasound and the clinical stages of ASO. Conclusion The overall classification of ASO on color Doppler ultrasound has considered impact of many other factors on the clinical symptoms，such as the level of the local narrow，narrow scope，segments of occlusion and collateral arteries，which divids the lesions more objectively，shows good consistency with the clinical stages.

Abstract:Objective To investigate the mediator effects of coping styles on the relationship between multidimensional life satisfaction (family,friends,school,living environment,self and global) and depression/anxiety,and  clarify the importance of coping strategies and their relationships with well being in the context of ongoing depression and anxiety. Methods The participants were 17 622 students of eight cities in China. The data were collected by using Multidimensional Students’ Life Satisfaction Scale (MSLSS),Trait Coping Style Questionnaire (TCSQ),Self-rating Depression Scale (SDS),and Self-Rating Anxiety Scale (SAS). Results Global life satisfaction and positive coping were negativiely  correlated with anxiety and depression（r=－0.471,－0.423,－0.550,－0.369,Ps< 0.01);negative coping was positively correlated with anxiety(r=0.369,P<0.01). The proportions of mediating effects in total effects of positive/ negative coping in associations among life satisfaction and depression were 35.4%-54.6%,9.6%-19.3%,respectively. The proportions of mediating effects in total effects of positive/ negative coping in associations amongst life satisfaction and anxiety were 16.4%-33.1%,29.0%-42.3%,respectively. Conclusion Coping styles partially mediate the relationship between multidimensional life satisfaction and depression/anxiety. It should pay more attention with regard to the study on the pathogenesis and prevention of  adolescent depression and anxiety.