Objective:To evaluate whether Salviae Miltiorrhizae Liguspyragine Hydrochloride and Glucose Injection has posi-tive effects on the prevention and treatment of radiation-induced lung injury. The basic function and mechanism of Salviae Miltiorrhi-zae Liguspyragine Hydrochloride and Glucose Injection were also investigated. Methods:A total of 70 adult male rats weighing about 200 g were selected and divided into seven groups. These groups were as follows:1) normal control group (N):rats were injected with 1 ml of normal saline per day;2) single medicine treatment group (D):rats were administered with 1 ml of Salviae Miltiorrhizae Ligus-pyragine Hydrochloride and Glucose Injection per day;3) single irradiation group (Z):rats were exposed to 20 Gy single whole-chest ir-radiation and injected with 1 ml of normal saline per day;4) irradiation with 10 Gy and medicine treatment group (Z10):rats were ex-posed to 10 Gy single whole-chest irradiation and administered with 1 ml of Salviae Miltiorrhizae Liguspyragine Hydrochloride and Glucose Injection per day; 5) irradiation with 15 Gy and medicine treatment group (Z15): rats were exposed to 15 Gy single whole-chest irradiation and administered with 1 ml of Salviae Miltiorrhizae Liguspyragine Hydrochloride and Glucose Injection per day;6) irradiation with 20 Gy and medicine treatment group (Z20a):rats were exposed to 20 Gy single whole-chest irradiation for four weeks, and administered with 1 ml of Salviae Miltiorrhizae Liguspyragine Hydrochloride and Glucose Injection per day;and 7) irradia-tion with 20 Gy and medicine treatment group (Z20b):rats were exposed to 20 Gy single whole-chest irradiation and administered with 1 ml of Salviae Miltiorrhizae Liguspyragine Hydrochloride and Glucose Injection per day. Two rats were selected and sacrificed at the end of two, four, six, eight, and ten weeks of irradiation. Samples of blood and lung tissues in rats were obtained. Results:In the group with single irradiation, the tumor necrosis factor-α (TNF-α) and transforming growth factor-β (TGF-β) levels in plasma of rats were higher than those in the other groups. In the group with irradiation and medicine treatment, the TNF-αand TGF-βlevels in plasma were higher than those in the normal control group and single medicine treatment group. In the group with single irradiation for four weeks,some petechial hemorrhages on the surface of the lung were visible to the naked eye. In the groups with medicine treatment, the petechi-al hemorrhages on the surface of the lung visibly reduced. According to the pathological mechanism of lung tissues, the groups with Sal-viae Miltiorrhizae Liguspyragine Hydrochloride and Glucose Injection exhibited less inflammation than the single irradiation group. Ir-radiation at 20 Gy for four weeks followed by a daily abdominal injection was slightly better than single irradiation, but the effects were not obvious. Conclusion: Salviae Miltiorrhizae Liguspyragine Hydrochloride and Glucose Injection could prevent the occurrence of lung injury by reducing the TNF-αand TGF-βlevels in plasma. After the occurrence of radiation-induced pneumonitis, the application of medicine could not decrease the symptoms.

Objective To investigate the relationship between the expressions of P16 and proliferating cell nuclear antigen(PCNA) in osteosarcoma and clinicopathological features,and explore the effects of them in occurrence and development of osteosarcoma.Methods The expressions of P16 and PCNA were detected by immunohistochemistry(SP) in 71 osteosarcima tissues and 10 normal bone tissues.Results ①The positive rate of P16 expression in osteosarcoma tissues was lower than that in normal bone tissues(P0.05).The expression of PCNA had positive relationship with osteosarcoma's prognosis(P0.05).③There was negative correlation between P16 and PCNA expressions(rs=-0.58,P

Objective:To investigate the expression of cyclooxygenase-2 in primary hepatocellular carcinoma, in cancer surrounding tissues and normal liver tissue and its relationship with clinical pathological features.Methods:The expression of COX-2 was detected in 30 cases of hepatocellular carcinoma, 20 cases of cancer surrounding tissue and 10 normal liver tissue by flow cytometry (FCM) and immunohistochemistry (SP). The clinical data were analyzed retrospectively.Results:(1)The expression of COX-2 in the HCC tissue was significantly higher than in cancer surrounding tissues and normal liver tissue (P0.05).Conclusion:The hyperexpression of COX-2 in tissue can reflex the biological behavior of HCC,and have very important role in the development of HCC.The specificness of COX-2 protein expression make it to be new target of tumor diagnosis and treatment.These results provide a theoretical basis for the chemoprevention of hepatoma.

Objective:To study the expression of COX-2 and Survivin in HCC tissues and their relationship for supplying experimental evidence for gene diagnosis and treatment of HCC.Methods:The expression of COX-2 and Survivin was detected in 30 cases of hepatocellular carcinoma and 10 normal liver tissue by Flow cytometry (FCM) and immunohistochemistry (SP).Results:Two experimental methods showed that the positive expression of COX-2 and Survivin in HCC was significanly higher than that in normal liver tissue and there was significanly positive correlation between the expression of COX-2 and Survivin.Conclusion:The hyper-expression of COX-2 and Survivin in tissue can reflex the biological behavior of HCC and there are synergistic effect between them during the development of HCC.A possible mechanism is inhibition of tumor cell apoptosis through upregulating Survivin by COX-2,and promoting abnormal cell proliferation in development of hepatocellular carcinoma.

Objective To explore an effective evaluation method for students' self-and peer-assess-ment. Methods The students of 6 groups participating in extracurricular teaching activities were selected as research subject. Traditional method (final score = mean score of group/2 + teacher's score/2) and mean difference method [final score=teacher's score-(mean difference of group-mean difference of all groups)] were used to calculate final score of each group, and effect of two methods were compared. Results Scores of most groups were higher than the teacher's scores, and high scores were given by group 3 in self- and peer-assessment. The final score of all groups were higher than teacher's scores in traditional method. Compared with teacher's scores, final scores increased significantly in group 1, 4, 5 below mean difference, final score decreased significantly in group 2, 3 above mean difference, and final score did not differ in group 6 equal to mean difference in mean difference method. Conclusion The mean difference method can reflect the effect of student's self- and peer-assessment, and guide student to make objective and accurate evaluation. It is a more reasonable and scientific evaluation method for self-and peer-assessment.

Objective To investigate the effect of enteral nutrition (EN) on the T lymphocytes-mediated immune function in patients with acquired immune deficiency syndrome (AIDS). Methods Totally 79 AIDS patients were randomly divided into enteral nutrition ( EN ) group ( supported with EN daily in addition to conventional treatment; n = 46) and control group (underwent conventional treatment only; n = 33 ). T lymphocytes including CD3, CD4, and CD8 cells as well as blood biochemical parameters including alanine aminotransferase ( ALT), aspartate aminotransferase (AST), glucose ( Glu ), total protein (TP), albumin ( ALB ), blood urea nitrogen (BUN) , Cr, and prealbumin (PA) were determined immediately before management (T0) and on the 30th day(T1). Results ALT, AST, Glu, TP, ALB, BUN, Cr, and PA showed no significant differences between these two groups before management ( all P ＞ 0. 05 ). The levels of TP ( P = 0. 015), ALB ( P = 0. 007 ), and PA ( P =0. 022 ) were significantly higher in EN group than those in control group at T1. The cell counts of CD3, CD4, and CD8 were not significantly different at T0, while the cell count of CD4 was significantly higher in EN group than that in control group at T1 ( P ＜ 0. 05 ). Conclusion EN can improve the nutritional status and T lymphocytesmediated immune function in AIDS patients.

BACKGROUND:Embryonic stem cells (ESCs) serve as a major cell source for smooth muscle cells,but the heterogeneity of cells derived from ESCs result in difficulty to obtain high purity smooth muscle cells.OBJECTIVE:To construct a double expression vector of puromycin resistance (pac) gene and enhanced green fluorescence protein (EGFP) gene driven by smooth muscle specific SM22α promoter (pSM22α-PAC-IRES2-EGFP),in addition,to detect its availability and specificity in ESCs.DESIGN,TIME AND SETTING:The observational experiment of gene level was performed at the Cardiovascular Institute,General Hospital of Shenyang Military Region from April 2007 to September 2008.MATERIALS:ESCs line R1 with number SCRC-1011TM was purchased from American ATCC Company.The pSM22α-EGFP vector was constructed by our laboratory.And the pIRES2-EGFP,pSM2C and pSuper.basic vectors were purchased from Invitrogen Company.METHODS:SM22α promoter was cloned from pSM22α-EGFP by polymerase chain reaction.CMV promoter of pIRES2-EGFP vector was replaced by SM22 promoter to establish pSM22α-IRES2-EGFP.Pac gene,excised from pSM2C by HindⅢ/Clal digestion,was sub-cloned into pSuper.basic to establish pSuper-PAC.After BgⅢ/Accl enzyme digestion of pSuper-PAC,pac gene fragment was obtained,which was further sub-cloned into pSM22α-IRES2-EGFP to produce pSM22α-PAC-IRES2-EGFP.ESCs were transfected with pSM22α-PAC-IRES2-EGFP using lipofectamine.Positive clones were selected by G418 and induced to differentiate and further identified by amplification of pac gene by RT-PCR.Differentiated cells were immunostained by SM α-actin,and expression of SM α-actin and EGFP was observed simultaneously under fluorescence microscope.MAIN OUTCOME MEASURES:Sequencing result of pSM22α-PAC-IRES2-EGFP;Amplification of pac gene;EGFP expression;as well as SM α-actin immunostaining.RESULTS:Three segments of 261 bp,664 bp,and 5000 bp were obtained by HindⅢ/Clal digestion,which was coincident with expectation,and the sequencing results showed that pSM22α-PAC-IRES2-EGFP vector was successfully constructed.Amplification of pac gene identified 4 ESCs clones successfully transfected.After induction of differentiation,partial portion of differentiated cells expressed EGFP,accompanied by positively stained by SM α-actin antibody.CONCLUSION:pSM22α-PAC-IRES2-EGFP vector was successfully constructed.ESCs clones transfected with this vector expressed pac gene and EGFP gene,and the expression of EGFP is smooth muscle specific.

Objective:To investigate mechanism of TRAIL-resistance in A549 cells ( a cell line of non-small cell lung carcino-ma cells) due to Akt phosphorylation .Methods:A549 cells were treated with Akt inhibitor Perifosine and rsTRAIL protein individual-ly and in combination.The expressions of Akt phosphorylation(p-Akt),c-FLIPLL and caspase-8 were detected by Western blot.The apoptotic rate of the A549 cells treated was detected by flow cytometry and the cell proliferation was evaluated by MTT assay .Results:A549 cells showed the increased level of Akt phosphorylation mediated by rsTRAIL protein .Treatment with the Akt inhibitor Perifosine induced a suppression of Akt activation in A 549 cells and a concomitant decrease in the expression of c-FLIPLL .As a result, Perifosine significantly enhanced TRAIL-induced apoptosis rate of (76.5 ±3.02)%and cytotoxic rate of (83.2 ±2.54)%by promoting the activ-ity of caspase-8.Conclusion:Akt activity promotes A549cells survival against TRAIL-induced apoptosis and that the cytotoxic effect of rsTRAIL protein can be enhanced by modulating the Akt phosphorylation in human non -small cell lung carcinoma cells .

Objective To study the influence of survivin targetedly inhibited with antisense oligonucleotide (ASODN)technique on the apoptosis of hepatoma carcinoma cells SMMC-7221,and to clarify the mechanism of promotion effect of survivin-ASODN on the apoptosis of SMMC-7721 cells.Methods The ASODN sequence of survivin marked by FAM fluorescein was designed and synthized. The SMMC-7721 cells were transfected by different concentrations (100,200,300,400,and 600 nmol· L-1 )of survivin-ASODN (ASODN transfection groups),at the same time blank control group and blank liposome control group and sense oligonucleotide (SODN) control group were set up.The apoptotic rates and the changes of cell cycle of the SMMC-7721 cells 24,48,and 72 h after transfected with different concentrations of survivin-ASODN were detected by FCM. The expression levels of survivin were measured by Western blotting method.Results Compared with each control group,24 h after transfection,the apoptotic rates of survivin-ASODN transfected SMMC-7221 cells were increased,the growth of cells was inhibited (P<0.05),and the effects had time-dose dependent tendency.48 h after transfection,the hypodiploid apoptotic peak appeared in ASODN transfection groups before G1 phase, the number of the cells at G0/G1 phase was decreased (P<0.05)and the number of the cells at G2/M phase wsa increased (P<0.05). Compared with each control group,the survivin expression levels in the SMMC-7721 cells in ASODN transfection groups were decreased (P<0.05 ), and the effects of survivin-ASODN was time-dose dependent (P<0.05 ). Conclusion Survivin-ASODN can block the expression of survivin in SMMC-7721 cells and inhibit the proliferation of SMMC-7721 cells by changing the cell cycle and increasing apoptosis in a time-dose dependent manner.

Objective To explore the potential mechanisms of non-small cell lung carcinoma cells to rsTRAIL protein-induced apoptosis by phosphatidylinositol 3′-kinase (PI3K/Akt)inhibitor LY294002,and to provide new ways to increase killing activities of rsTRAIL protein for non-small cell lung cancer.Methods The A549 cells at logarithmic growth phase were selected and randomly divided into rsTRAIL group and LY294002+rsTRAIL group. The inhibitory rate of growth of the A549 cells was tested by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry analysis. The expression levels of Ser473 phosphorylated form of Akt (p-Akt),c-FLIPL protein and Bcl-2 protein in the A549 cells in two groups were analyzed by Western blotting method. Results The inhibitory rate of growth of the A549 cells in LY294002+rsTRAIL group (74.6 %± 2.63%)was higher than that in rsTRAIL group (5.61% ± 0.32%) (P< 0.05 ). Compared with rsTRAIL group, the percentage of the cells at G0/G1 phase in LY294002+rsTRAIL group was increased(P<0.05)and the percentage of the cells at S phase was decreased(P<0.05).The apoptotic rate of the A549 cells in LY294002+rsTRAIL group (61.5%±3.02%)was higher than that in rsTRAIL group (3.21%±0.96%)(P<0.05). The Western blotting results showed that the expression levels of p-Akt, c-FLIPL and Bcl-2 proteins in the A549 cells in LY294002+rsTRAIL group were decreased (P<0.05 )and the ratio of Bax/Bcl-2 was increased (P<0.05 ) compared with rsTRAIL group.Conclusion LY294002 can increase the killing activity of rsTRAIL protein in A549 cells by inhibiting the activity of PI3K.