Objective To investigate the role of mitochondrial KATP channel in the mechanism of the protective effect of ischemic preconditioning (IP) against ischemia-reperfusion (I/R) injury.Methods Forty-eight Wistar rats of both sexs weighing 250-350 g were used in this study. Forty rats were randomly divided into 5 groups ( n = 8 each): group A I/R; group B IP+ I/R; group C diazoxide (DZ mito-KATP channel activator) + I/R; group D 5-HD (mito-KATP channel blocker) + IP + I/R and group E 5-HD + DZ + I/R. Another 8 animals were used for electron microscopic examination of normal mitochondria as control. The animals were anesthetized with intraperitoneal pentobarbital 30 mg?kg-1. The hearts were immediately excised and passively perfused in a Langendorff apparatus with K-H solution at 5.8 kPa perfusion pressure and 36.5-37.5℃ via aortic cannulation. A fluid-filled latex balloon was via left atrium in left ventricle for the measurement of left ventricular function. I/R was induced after 30 min stabilization by clamping aortic cannula for 40 min followed by 30 min reperfusion. In group B and D the isolated hearts underwent 2 episodes of 5 min ischemia followed by 5 min reperfusion before I/R. In group C and E DZ 50 ?mol?L-1 was infused for 10 min and in group D and E 5-HD 100 ?mol?L-1 was infused for 10 min before I/R. HR, LVSP, LVEDP and coronary flow (CF) were measured at the end of stabilization (T0 , baseline), immediately before I/R (T1 ) and at 10, 20 and 30 min of reperfusion (T2.3.4.), and left ventricular developed pressure (LVDP= LVSP- LVEDP) was calculated. Myocardial tissue was obtained at the end of 30 min reperfusion for electron microscopic examination of mitochondria. Mitochondrial ultrastructure was assessed by Flameng scoring system (0 = normal, 4 = severely damaged) .Results Ischemic and DZ preconditioning significantly increased LVDP and decreased LVEDP and Flameng score. 5-HD pretreatment partly antagonized the protective effect of IP and completely antagonized that of DZ against I/R injury. Conclusion Ischemic preconditioning protects the heart against I/R injury mainly by activating mitochondrial KATP channel.

Objective To establish a cell line of human ovarian cancer, and study its characterization. Methods The cell line was established by the cultivation of subsides walls, and kept by freezing. The morphology was observed by microscope and electromicroscope. The authors studied its growth and propagation, the agglutination test of phytohemagglutinin (PHA), the chromosome analysis, heterotransplanting, immuno histochemistry staining, the analysis of hormone, the pollution examination and the test of sensitivity to virus etc. Results A new human ovarian carcinoma cell line, designated ovarian mucinous cystadenocarcinoma 685 (OMC685), was established from mucinous cystadenocarcinoma. This cell line had subcultured to 91 generations, and some had been frozen for 8 years and revived, still grew well. This cell line possessed the feature of glandular epithelium cancer cell. The cells grew exuberantly, and the agglutinating test of PHA was positive. Karyotype was subtriploid with distortion. Heterotransplantations,alcian blue periobic acid schiff (AbPAS),mucicarmine,alcian blue stainings, estradiol (E 2) and progesterone were all positive. Without being polluted, it was sensitive to poliovirus Ⅰ, adenovirus 7 and measles virus. Conclusions OMC685 is a distinct human ovarian tumous cell line.

Objective To investigate the effects of Tet-on gene regulated HIF-1?expression on cell apoptosis of hepatoma cells in vitro. Methods The apoptosis of hepatocellular carcinoma cell lines HePG2 was measured after HIF-1?expression was activated in HePG2 in vitro by Tet-on expression system. Results The amplified products were confirmed as the cDNA of HIF-1?by DNA sequencing, and obtained pTHE-HIF-1?was verified by endonuclease digestion. The HePG2Tet-on cells express HIF-1?constantly. After being incubated under different concentrations of doxycycline for 48 h, HIF-1?gene could inhibit HePG2 cell apoptosis. The apoptosis rates were 59. 6%、50. 9%、38. 1%、30. 5%、23. 9%、18. 3% respectively when the concentrations of doxycycline were 0?g/ml,0. 02?g/ml,0. 2?g/ml、1?g/ml、2?g/ml,5?g/ml (P

Objective To examine whether the increase of carbon monoxide (CO) induced by oral methylene chloride (MC) administration in recipients before heart transplantation would protect heart grafts against isehemia/reperfusion (I/R) injury associated with transplantation and to explore the possible mechanism.Methods Inbred male Balb/c mice were used as donors and recipients to establish cervical heart transplantation model Recipients were treated with either MC (100 mg/kg or 500 mg/kg,per os)(group MC 100 mg,n=10;group MC 500 mg,n=12) or olive oil(0.15 ml,per os.group olive,n=10) 3 h prior to anesthesia.Age-matched norwlal mice served as controls (group N,n=5).The serum COHb and the CO content of myocardial tissue were measured at 0,1,3,6,12,24 h after oral MC administration.Half of recipients were killed at 3 and 24h after transplantation for senum or cardiac graft samples.The serum cTnI levels,the mRNA levels of TNF-α,IL-10,Bcl-2,Bax.the protein levels of NF-κB and the ultrastructures of myocardium were examined.Results As tompared with group olive.the serum COHb and tissue CO were increased significantly and peaked within 3 h in group MC 100 mg and group MC 500 mg.The serum cTnI levels in group MC 100 mg and group MC 500 mg were significantly decreased (P<0. 01 ), especially in group MC 500 mg. The increase of CO in recipients of group MC100 mg and group MC 500 mg significantly inhibited the proinflammatory gene expression of TNF-α mRNA and the pro-apoptotic gene expression of Bax mRNA (P<0. 01), and increased the anti-apoptotic gene expression of Bcl-2 mRNA (P<0. 01), but did not increase the anti-inflammatory gene expression of IL-10 mRNA (P>0. 05) in the heart grafts. As compared with group N, the myocardial NF-κB activation was increased significantly in group olive,group MC 100 mg and group MC 500 mg (P<0. 01 ), but there was no significant difference among the later three groups (P>0. 05). The myocardial ultrastructure was also alleviated significantly in group MC 100 mg and group MC 500 mg as compared with group N. Conclusion The increase of CO induced by MC in recipients suppresses pro-inflammatory and pro-apoptotic gene expression and efficiently ameliorates transplant-induced heart I/R injury. The possible mechanism does not seem to be associated with down-regulation of the NF-κB signaling pathway.

Objective To investigate the role of induction of carbon monoxide (CO) with methylene chloride (MC) in recipients in ameliorating allograft rejection and prolonging allograft survival and to explore the possible mechanisms in a murine heart transplantation model.Methods Inbred male C57BL/6 mice were used as donors and inbred male Balb/c mice as recipients respectively to establish cervical heterotopic heart transplantation model.The experiments were divided into 3 groups.Recipients were treated with MC (100 mg/kg,per os) day 1 prior to transplantation to day 6 posttransplantation (group MC1w) or to day 13 posttransplantation (group MC2w),or treated with isovolumic olive oil day 1 prior to transplantation to cardiac arrest of allograft (group Tx).The serum TNF-α and IL-10 proteins,TNF-α and IL-10 mRNA,and Foxp3 mRNA and protein in cardiac grafts were measured respectively.The intercellular adhesion molecule-1 (ICAM-1) and Caspase-3 protein in cardiac grafts,and the histopathologic changes of cardiac grafts were also observed.Results Serum COHb levels in untreated mice were (0.85 ± 0.28)％.After MC application,serum COHb peaked within 3 h in recipients (5.24 ± 0,45)％ (P＜0.01 ).The median survival time of cardiac grafts in group MC1w(12.1 days) and group MC2w( 19.4 days) was longer than that in group Tx (6.3 days) (P ＜0.01). As compared with group Tx,induction of CO in group MC1w and group MC2w down-regulated significantly the levels of serum TNF-α (P＜0.01 ) and TNF-α mRNA (P＜0.01) of cardiac grafts and spleen in recipient mice,inhibited the protein expression of ICAM-1 (P＜0.01) and Caspase-3 (P＜0.01) of cardiac grafts,and inhibited,especially in group MC2w,the proliferation of lymphocytes and monocytes infiltration in cardiac grafts.There was no significant difference in serum IL-10 and Foxp3 mRNA and protein in cardiac grafts and spleen of recipients among the groups (P＞0.05).Conclusion Induction of CM in recipients could relieve cardiac allograft rejection and prolong cardiac allograft survival via its anti-inflammation and anti-apoptotic effects,not via up-regulation of Foxp3 in recipient mice.

Objective To study the effect of three fungi of Helminthosporium on the seven kinds of bioactive constituents in root of seedling Rumex gmelini. Methods Let the R. gmelini infect the fungi of Helminthosporium by dealing the leave surface of R. gmelini with fungal spore suspension. The content of seven kinds of bioactive constituents was determined by HPLC and the results were analyzed by variance analysis. Results During the treatment time, little effect of three fungi of Helminthosporium on content of polydatin, chrysophanol 1-glucoside, and physcion, but the obvious effect on resveratrol, musizin, emodin, and chrysophanol was found. Conclusion The effects of H. turcicum 001 on yield and content of resveratrol are significant in early-middle stage (0—40 d), but treatment time over 40 d is not suitable. The results of this study could provide the new idea and theoretical basis for the exploiting of natural medicines and the planting of Chinese herbal medicine.

Objective:To explore the hemodynamic and comfort effect of modified prone-flexed position in complexity renal calculi patients undergoing percutaneous nephrolithotomy.Methods:A total of 82 cases of complexity renal calculi patients who undergoing percutaneous nephrolithotomy were recruited and randomly assigned to experiment group (41 cases) and control group (41 cases). Patients in the control group were placed in conventional prone position, while in the experiment group were placed in modified prone-flexed position. The data include blood pressure, heart rate, respiratory rate, saturation of blood oxygen were recorded at preoprative, lithotomy position, change position 30 minutes after operation and postoperative. The General Comfort Questionnaire (GCQ) score was recorded in patients for postural comfort. The clinical data was also compared between two groups.Results:There was no significant difference in blood pressure, heart rate, respiratory rate, saturation of blood oxygen between experiment group and control group ( P>0.05). Compared to (44.95±4.80) min, 90.2% (37/41) of the control group, the patients in the experiment group was (47.80±6.71) min, 73.2% (30/41), which showed a shorter average operation time and higher total stone clearance rate, the differences were statically significant ( tvalue was 2.216, χ2value was 3.998, P<0.05). At immediately and 30 minutes after operation, the skin pressure were 9 cases and 4 cases in the experiment group, 18 cases and 11 cases in the control group, the differences were statically significant ( Zvalues were 2.132, 2.038, P<0.05). After 48 hours of surgery, the physical subscale and totl GCQ score were (13.32±1.40) and (78.56±4.65) pionts in the experiment group, significantly higher than (12.17±1.41), (75.76±5.16) pionts in the control group, the differences were statically significant ( tvalues were 3.685, 2.584, P<0.01 or 0.05). Conclusion:Modified prone-flexed position can shortern operation time decrease the risk of pressure injury and demonstrate better postural comfort of patients with complexity renal calculi undergoing percutaneous nephrolithotomy.

0. 05). At the protein level, the expressions of MDR1, BCRP and LRP in HePG2/ADM were significantly higher than HePG2 and L02 ( FMDR1= 28. 68, FBCRP= 18. 60, FLRP= 6. 28, P 0. 05). Conclusion The four genes are probably the normally expressed genes in the liver. The expression of MDR1 and BCRP could be up-regulated by anticancer agents in vitro. MDR1 and BCRP may play very important roles in the MDR of HePG2/ADM. However, MRP1 and LRP may have no relations with the multi-drug resistance of HePG2/ ADM cell lines.

OBJECTIVE:To provide references for the application and management of anti-infectives.METHODS:Data on sale volume,DDDs etc.of anti-infection drugs in6hospitals of Guang'an city from2001to2004were statistically analyzed.RESULTS:The DDDs of oral lomefloxacin dominated the first or2nd places for the4years;The DDDs rank orders of isoniazid and rifampin were No.4and No.5respectively in the4years;The DDDs of penicillin for injection dominated the first place on the injectable preparation lists(2001～2003);The latecomer levofloxacin assumed the tendency of surpassing the early starters not only in sale volume but also the DDDs.CONCLUSION:The proportion for anti-infectives in sales volume to the sum total drug consumption of the above mentioned area were close to those of the other medical institutions in southwest area,there are less new drug varieties and the grade of drugs used in the area was low.

The roles of multi-drug resistance protein 1 (MDR1), multi-drug resistance related protein 1 (MRP1), lung resistance protein (LRP) and breast cancer resistance protein (BCRP) in the multi-drug resistance (MDR) of hepatocellular carcinoma (HCC) were studied. By exposing HepG2 cell line to progressively increased concentrations of adriamycin (ADM), HepG2 multi-drug resistant subline (HepG2/ADM) was induced. The MDR index of HepG2/ADM was detected by using MTT. The expressions of the four MDR proteins in the three cell lines (L02, HepG2, HepG2/ADM) were investigated at mRNA and protein levels by real-time RT-PCR and Western blot respectively. Our results showed that when the ADM concentration was under 100 microg/L, HepG2 could easily be induced to be drug-resistant. The IC(50) of the HepG2/ADM to ADM was 282 times that of HepG2. The expression of MDR1 and BCRP mRNA in HepG2/ADM cells were 400 and 9 times that of HepG2 cells respectively while there was no difference in the mRNA expressions of MRP1 and LRP. There was no difference between HepG2 and L02 cells in the mRNA expressions of the four genes. At the protein level, the expressions of MDR1, BCRP and LRP but MRP1 in HepG2/ADM were significantly higher than those of HepG2 and L02. Between HepG2 and L02, there was no difference in the expressions of four genes at the protein level. HepG2/ADM is a good model for the study of MDR. The four genes are probably the normally expressed gene in liver. The expressions of MDR1 and BCRP could be up-regulated by anti-cancer agents in vitro. The MDR of HCC was mainly due to the up-regulation of MDR1 and BCRP but MRP1 and LRP. These findings suggest they may serve as targets for the reversal of MDR of HCC.