Objective To evaluate a new computer-aided technique applicable for myocardial contrast echocardiography(MCE) to quantitate automatically calibrated myocardial contrast intensity(CD and to test the feasibility of calibrated CI in assessing myocardial perfusion. To analyze the relationship on myocardial perfusion,regional contractile function and cell apoptosis in stunned myocardium. Methods According to coronary occlusion and reperfusion at different times, rabbits were divided into three groups: 15 min occlusion/30min reperfusion (group Ⅰ ),30 min occlusion / 60min reperfusion (group Ⅱ ) and 120 min occlusion / 60min reperfusion (group Ⅲ ). MCE was performed on all rabbits at baseline,occlusion and after reperfusion,and its images were analyzed by a new computer-aided technique. Myocardial calibrated CI of each segment was measured automatically by software. Percentage wall thickening (WT) of each risk segment at each stage were also measured by echocardiography. The apoptotic index(AI) in regional left myocardial dysfunction was calculated by terminal deoxynucleotidyl transferease-mediated biotinylated deoxyuridine triphosphate nick end labeling(TUNEL ). Results (1) During occlusion, WT in the areas at risk decreased to zero or negative and the calibrated CI values were significantly lower than those at baseline ( P ＜0.01 ). After reperfusion, WT in all risk segment remained depressed, but calibrated CI significantly improved in group Ⅰ and Ⅱ while those remained unchanged in group Ⅲ. (2)AI in risk myocytes were (13. 70 ± 5.48 ) %, (36.25 ± 5.55 ) % and ( 62.06 ± 6. 70 ) %, respectively, both statistically significant difference between the two groups ( P ＜0.05 or P ＜ 0.01 ). AI were negatively correlated to WT and calibrated CI ( r = - 0. 87 and r = - 0. 77, P ＜0.05). Conclusions MCE with computer-aided technique can assess quantitatively myocardial perfusion and regional contractile function. Short-term ischemiareperfusion does not cause myocardial necrosis, but it will lead to myocardial cell apoptosis and the phenomenon of myocardial stunning. Prolonged ischemia, even if given sufficient reperfusion, can lead to apoptosis and necrosis simultaneously.

20 first mandibular premolars were randomly divided into 2 groups(n =10).The teeth in experimental group were treated by fi-ber main post in combination with auxiliary pile,in control group by single fiber main post,and then were restored by metal crown.They were fixed in universal testing machine.The fracture load(N)of experimental and control group was (846.50 ±40.33)and (437.90 ±41.15) respectively(P <0.01).

Objective To evaluate the effects of dexmedetomidine (DEX) on cell apoptosis induced by endoplasmic reticulum stress and c-Jun N-terminal kinase (JNK) pathway during one-lung ventilation (OLV) in rats.Methods Sixty male Sprague-Dawley rats were randomly allocated into 6 groups (n =10 each):sham operation group (Sham group),OLV group,OLV + atipamezole (α2 receptor antagonist) group (AD group),OLV + atipamezole + DEX group (DEX+AD group),OLV + low-dose DEX group (DEX-L group) and OLV + high-dose DEX group (DEX-H group).The animals were anesthetized with 10％ chloral hydrate 4.5 ml/kg,tracheostomized and mechanically ventilated.Bilateral lungs were ventilated for 2.5 h in Sham group.The right lung was ventilated for 2.0 h followed by 0.5 h two-lung ventilation in OLV group.In DEX-L and DEX-H groups,DEX was infused intravenously for 1 h at a rate of 2.5 μg · kg-1 · h-1 and 5.0 μg · kg-1 · h-1,respectively,starting from 1 h prior to OLV.Atipamezole 250 μg/kg was injected intravenously at 1 h prior to OLV in AD group.Atipamezole 250 μg/kg was injected intravenously at the onset of DEX infusion (5.0 μg · kg-1 · h-1) in DEX+AD group.The rats were sacrificed and left lungs were removed for determination of weight to dry lung weight ratio (W/D),cell apoptosis in lung tissues (by TUNEL),and expression of glucose-regulated protein 78 (GRP78) mRNA and protein,JNK mRNA and phosphorylated JNK (p-JNK) protein (by RT-PCR and Western blot).Pathological changes of lungs were examined and the injured alveolus rate (IAR) was counted under light microscope.The changes in ultrastructure of lung tissues were observed under transmission electron microscope.Apoptosis index (AI) was calculated.Results W/D,AI and IAR were significantly higher in OLV,AD and DEX+AD group than in Sham group,while lower in DEX-L and DEX-H groups than in OLV,AD and DEX+AD groups.The pathological changes of the structure of lung tissues were observed in OLV,AD and DEX+AD groups,while the pathological changes were significantly alleviated in DEX-L and DEX-H groups.In OLV,AD and DEX + AD groups,there was apoptosis in lots of pulmonary vascular endothelial cells and alveolar epithelial cells,while cell apoptosis was significantly reduced after administration of DEX.The expression of GRP78 mRNA and protein,JNK mRNA and p-JNK protein was significantly higher in OLV,AD and DEX+AD groups than in Sham group,and lower in DEX-L and DEX-H groups than in OLV,AD and DEX +AD groups.Conclusion DEX pretreatment can protect lungs during OLV,and inhibited JNK signaling pathway and reduced cell apoptosis induced by endoplasmic reticulum stress may be involved in the mechanism.

Objective:To analyze and optimize the extraction technology of Geum aleppicum by Box-Behnken design and response surface methodology. Methods:With the content of total flavonoids as the index, the effects of three factors including the solvent-solid ratio, ethanol concentration and extraction time on the content of total flavonoids were studied by single factor experiment, and the ex-traction technology was optimized by Box-Behnken design and response surface methodology. Results: The optimal extraction condi-tions of Geum aleppicum were as follows:the solvent-solid ratio was 4. 2, the ethanol concentration was 50% and the extraction time was 80 min. Under the above conditions, the average yield was 12. 590 0 mg·g-1 . Conclusion: Optimizing extraction technology of the total flavonoids in Geum aleppicum by Box-Behnken design and response surface methodology is reasonable, and the mathematical mod-el is consistent with the experimental data, which has good predictability.

Objective To investigate a cure for neonatal hypoxic-ischemic encephalopathy(HIE) by observing the curative effect on neonates with HIE admitted to neonatal intensive care unit (NICU).Methods Fifty neonates with HIE from May to Dec.in 2006 admitted to NICU for treatment(NICU group),which contrast to the 42 neonates with HIE in corresponding periods of 2005 year(control group).The curative effect of three supports and three expectant treatment were evaluated by observing vital signs,oxygen saturation,biochemical test,and the neonatal beha-vioral nerve assessment (NBNA) respectively; Clinical curative effect contained markedly effective and effective which as the total curative effect statistics.Results There were no obvious difference between each group from the means of blood sugar data while hospitalizing as well as the NBNA in 3 days. By contrast, the blood sugar status and NBNA improving of neonates with HIE admitted to NICU surpassed the control group after treatment (Pa

BACKGROUND: Bone marrow mesenchymal stem cells are characterized by wide sources and low immunogenicity. Especially, these cells are easy to import and express foreign genes, and thus have obvious superiorities as anti-tumor gene therapy vectors. OBJECTIVE: To investigate the effects of interleukin-12 gene modified bone marrow mesenchymal stem cells on the proliferation, cell cycle and apoptosis of ovarian cancer cells. METHODS: Interleukin-12 recombinant adenovirus vector was used to transfect bone marrow mesenchymal stem cells. Then, the expression of interleukin-12 mRNA and protien in transfected bone marrow mesenchymal stem cells was detected by RT-PCR and western blot assay, respectively. The level of interleukin-12 in cell supernatant was determined by ELISA. The SKOV3 cells were co-cultured with the supernatant of bone marrow mesenchymal stem cells transfected with (transfection group) or without (control group) interleukin-12 recombinant adenovirus vector. The proliferation of SKOV3 cells was determined by MTT assay. Flow cytometry was used to detect the cell cycle and apoptosis of SKOV3 cells. RESULTS AND CONCLUSION: RT-PCR and western blot results showed that interleukin-12 mRNA and protein were expressed in transfected bone marrow mesenchymal stem cells, but not found in empty vector group and blank control group. ELISA results showed that the content of interleukin-12 in the supernatant of bone marrow mesenchymal stem cells was (68.78±12.35) μg/L in the interleukin-12 transfection group after 48 hours culture, and no interleukin-12 expression was detected in the empty vector group and the blank control group, Interleukin-12-transfected bone marrow mesenchymal stem cells significantly inhibited the proliferation of SKOV3 cells, and the proliferation inhibition rate was increased with time (P < 0.05). The proportion of G1-phased SKOV3 cells was higher in the transfection group than in the control group (P < 0.05), and the percentage of G2-phased SKOV3 cells was lower in the transfection group than in the control group (P < 0.05). The apoptosis rate of SKOV3 cells in the transfection group was higher than that in the control group (P < 0.05). Our experimental findings indicate that bone marrow mesenchymal stem cells transfected with interleukin-12 recombinant adenovirus vector can express interleukin-12, inhibit the proliferation and induce apoptosis of ovarian cancer cells.

BACKGROUND:Gene engineering plays an important role in the process of human malignant tumor treatment, and adenovirus vectors for gene therapy are commonly used. In recent years, lactoferrin is found to exert important effects on the occurrence, development and metastasis of a variety of tumors. However, little is reported on the in vivo expression of adenovirus vector mediated lactoferrin in the human body.OBJECTIVE:To investigate the effect of adenovirus mediated human lactoferrin (Ad-hLF) on proliferation and apoptosis of cervical cancer stem-like cells.METHODS:Primary cervical cancer cells from mice were cultured in vitro to sort cervical cancer stem-like cells using SP method. Afterwards, the stem-like cells were divided into three groups, blank control group, empty vector group, and Ad-hLF group, followed by transfection with nothing, Ad-GFP and Ad-hLF, respectively. After transfection, MTT assay was used to detect the proliferation of cervical cancer stem-like cells; scratch-wound assay was employed to detect the migration of cervical cancer stem-like cells; western blot assay was used to determine the expression of lactoferrin in cervical cancer stem-like cells; and Annexin V-FITC/PI was used to detect cell apoptosis.RESULTS AND CONCLUSION:Compared with the blank control and empty vector groups, Ad-hLF significantly inhibited the proliferation of cervical cancer stem-like cells and reduced the number of scratched cells, while the cell apoptosis increased in the Ad-hLF group (P < 0.05 or P < 0.01). These findings indicate that the recombinant adenovirus vector mediated lactoferrin Ad-hLF for transfection of cervical cancer stem-like cells can be stably expressed in cervical cancer stem-like cells, and can inhibit the proliferation, increase apoptosis and reduce migration of cervical cancer stem-like cells.

Objective To establish the HPLC characteristic chromatogram of Shenshitong Granules. Methods The chromatographie fingerprints were obtained through Thermo Hypersil GOLD-C18 column (4.6 mm×250 mm, 5μm) with the gradient elution solvent system composed of acetonitrile-0.2% phosphoric acid (0-20 min, 5%→10% acetonitrile;20-40 min, 10%→12%acetonitrile;40-60 min, 12%→14%acetonitrile;60-90 min, 14%→20%acetonitrile;90-120 min, 20%→28%acetonitrile). The detective wavelength was set at 280 nm;the flow rate was 1.0 mL/min;the column temperature was maintained at 30 ℃;the analysis time was 120 min. Results The HPLC characteristic chromatogram was built on basis of 10 batches of Shenshitong Granules, including 27 common peaks which contain the characteristic peaks of 6 Chinese herbal medicines, such as Radix Salvia Miltiorrhizae, Herba Lysimachiae, etc. Conclusion The established HPLC fingerprint has high sensitivity and good repeatability, and can be available for quality evaluation of Shenshitong Granules.

Objective To evaluate the effect of exogenous hydrogen sulfide (H2S) on mitochondrion-dependent apoptosis in lung tissues of rats with endotoxemia.Methods Forty healthy male SpragueDawley rats,aged 2-3 months,weighing 250-310 g,were divided into 5 groups (n=8 each) using a random number table:control group (group C),lipopolysaccharide (LPS) group,low dose sodium hydrosulphide (NaHS) group (group L-NaHS),moderate dose NaHS group (group M-NaHS) and high dose NaHS group (group H-NaHS).Endotoxemia was induced by Ⅳ LPS 5 mg/kg in chloral hydrate-anesthetized rats.The equal volume of 0.9％ sodium chloride solution was intravenously injected in group C.NaHS (an exogenous donor of H2S) 0.78,1.56 and 3.12 mg/kg were intraperitoneally injected at 3 h after LPS injection in L-NaHS,M-NaHS and H-NaHS groups,respectively.The rats were sacrificed at 6 h after injection of LPS or 0.9％ sodium chloride solution,and lungs were removed for examination of the mitochondrial ultrastructure of lung tissues and for determination of apoptosis in lung cells (by flow cytometry) and expression of caspase-3,caspase-9,Bcl-2 and Bax protein and mRNA in lung tissues (by Western blot or real-timne polymerase chain reaction).The apoptosis rate and ratio of Bcl-2 expression to Bax expression (Bcl-2/Bax ratio) were calculated.The expression of cytochrome c (Cyt c) in cytoplasm and mitochondria of lung tissues was detected by Western blot.Results The apoptosis rate was significantly increased,the expression of caspase-3,caspase-9,Bcl-2 and Bax protein and mRNA was up-regulated,Bcl-2/Bax ratio was decreased,the expression of Cyt c in cytoplasm was up-regulated,and the expression of Cyt c in mitochondria was down-regulated in group LPS (P ＜0.05 or 0.01).Compared with group LPS,the apoptosis rate was significantly decreased,the expression of caspase-3 and Bax protein and mRNA was down-regulated,the expression of Bcl-2 protein and mRNA was up-regulated,Bcl-2/Bax ratio was increased,the expression of Cyt c in cytoplasm was down-regulated,and the expression of Cyt c in mitochondria was up-regulated in L-NaHS,M-NaHS and H-NaHS groups,the expression of caspase-9 protein and mRNA was significantly down-regulated in M-NaHS and H-NaHS groups,and the expression of caspase-9 was significantly down-regulated (P ＜0.05 or 0.01),and no significant change was found in caspase-9 mRNA expression in group L-NaHS (P＞0.05),and the damage to mitochondrial ultrastructure was significantly mitigated in MNaHS and H-NaHS groups.Conclusion The mechanism by which exogenous H2S inhibits cell apoptosis in lung tissues may be related to inhibition of mitochondrion-dependent apoptosis in rats with endotoxemia.

Objective To observe the effects of hydrogen sulfide (H2S) on structure and function of mitochondria of lung in rats with acute lung injury (ALI) induced by lipopolysaccharide (LPS).Methods Forty healthy male Sprague-Dawley (SD) rats were randomly divided into control group, LPS injury group, and low-, middle-, and high-dose NaHS groups, with 8 rats in each group. The rats in LPS injury group were given LPS 5 mg/kg via sublingual vein, and those in low-, middle-, and high-dose NaHS groups were challenged by LPS for 3 hours followed by intraperitoneally injection of 0.78, 1.56 and 3.12 mg/kg NaHS respectively in a volume of 2 mL/kg. The rats in control group were given 2 mL/kg normal saline via sublingual vein. The rats were sacrificed at 6 hours after model reproduction, and the lung tissues were harvested on time. The mitochondria in lung tissues were isolated with differential centrifugation. The lung mitochondria ultra structures were observed with electron microscope. The content ofmalondialdehyde (MDA) in mitochondria was determined with thiobarbituric acid method, and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and adenosine triphosphatase (ATPase) were determined with xanthine oxidase method. The mitochondrial activity and swelling were determined by multiskan spectrum.Results It was shown by transmission electron microscope that the mitochondrial structure in the control group was normal. The mitochondria in rat lung cells was swollen with disrupted or disintegrated cristae, the osmiophilic lamellar bodies had fused or disappeared, and rough endoplasmic reticulum degranulation phenomenon was obvious in LPS injury group. The mitochondrial damage was slightly mitigated in the low-dose NaHS group, and it was significantly mitigated in the middle-dose and high-dose NaHS groups. Compared with control group, the MDA content in lung mitochondria in LPS injury group was significantly increased (nmol/mg: 26.30±1.45 vs. 11.16±1.20), andSOD, GSH-Px, and ATPase activities were significantly decreased [SOD (U/mg): 18.78±1.13 vs. 27.44±1.97, GSH-Px (U/mg): 63.91±1.99 vs. 128.15±3.47, ATPase (U/mg): 4.83±0.25 vs. 9.92±0.65]; as well as the activity of the mitochondria was significantly decreased (A value: 0.164±0.025 vs. 0.319±0.045), and the swelling of the mitochondria was significantly increased (A value: 0.182±0.012 vs. 0.273±0.023), all with significantly statistical differences (allP < 0.01). Compared with LPS injury group, the MDA contents in low-, middle-, and high-dose NaHS groups were significantly decreased (nmol/mg: 21.89±1.23, 17.63±1.56, 12.19±1.30 vs. 26.30±1.45), and the SOD, GSH-PX, and ATPase activities were significantly increased [SOD (U/mg): 20.13±0.85, 21.38±1.22, 24.05±1.56 vs. 18.78±1.13; GSH-Px (U/mg): 82.06±1.65, 101.45±2.14, 117.80±2.12 vs. 63.91±1.99; ATPase (U/mg): 5.34±0.23, 7.06±0.37, 8.78±0.44 vs. 4.83±0.25]; as well as the activity of the mitochondria was markedly increased (A value: 0.194±0.018, 0.230±0.032, 0.297±0.038 vs. 0.164±0.025), and the swelling of mitochondria was markedly decreased (A value: 0.195±0.008, 0.219±0.017, 0.249±0.018 vs. 0.182±0.012), all with significantly statistical differences (allP < 0.05). Moreover, the protective effect of NaHS showed a dose-dependent manner.Conclusion It could be concluded that LPS induce mitochondrial structural damage and functional impairment in rats with ALI induced by LPS, and H2S have a beneficial effect against ALI induced by LPS with decreasing the mitochondrial lipid peroxidation level and protecting the cell structure and function, and the effect is correlated with the dosage.