Object To study on the genetic diversity and genetic structure of Iphigenia indica Kunth. Methods Random amplified polymorphic DNA (RAPD) was applied to detect DNA fingerprints of three populations of I. indicafrom Yunnan Province. Results Twenty primers were screened, and a total of 120 DNA fragments were amplified ranging from 0.2-3 kb, among which 71 (59.17%) were polymorphic. The average number of DNA band produced by each primer was 3.55. The Shannon index was (0.199 4) in Ca population, 0.200 7 in Cb population, 0.254 8 in Cc population, respectively; the average value of populations was 0.218 3. The Shannon index was 0.288 6 in species. Nei's genetic identity was 0.930 9 between Ca and Cb, 0.932 7 between Ca and Cc, 0.946 6 between Cb and Cc. G_(st) within population was (0.204 4.) Conclusion For the establishment of protective tactic and measure, and of the standard of good agriculture practice (GAP) growth and heredity breeding, RAPD analysis provides I. indicawith theoretical foundation and basal data.

Objective To investigate the levels of peripheral blood plasma M-ficolin/L-ficolin and their correlation with clinical indexes in patients with rheumatoid arthritis (RA).Methods M-ficolin/L-ficolin was detected in the plasma of 61 RA patients who were grouped according to the disease activity and 18 healthy controls by enzyme-linked immunosorbent assay (ELISA).The relationships between the levels of M-ficolin/L-ficolin and laboratory parameters,disease activity were investigated in RA patients.Statistical analysis were performed by t-test,one-way analysis of variance (ANOVA),rank-sum test,correlation analysis was carried out with Pearson/ Spearman rank correlation test.Results ① M-ficolin levels in 61 RA patients were [M (QR) 39 (24) ng/ml] significantly higher than healthy controls (14±4) ng/ml (Z=-5.72,P＜0.05).L-ficolin levels in 61 RA patients were (1.7±0.6) μg/ml obviously higher than healthy controls (1.3±0.5) μg/ml (t=2.55,P=0.013).② The M-ficolin levels of 61 RA patients in low,mediate,high disease activity group were (21±5) ng/ml,[M(QR) 35(13) ng/ml),[M(QR) 50(25) ng/ml].There was significant difference among the three groups (P＜0.01).The L-ficolin levels of 61 RA patients in low、mediate,high disease activity group were (1.2±0.5) μg/ml,(1.7±0.4) μg/ml and (2.2±0.6) μg/ml.There was significant difference among the three groups (P＜0.05).(M-ficolin levels were positively correlated with disease activity score (DAS)28,erythrocyte sedimentation rate (ESR),C reactive protein (CRP),rheumatoid factor (RF),white blood cell (WBC),Note Express User Tool (NEUT),MONO,blood platelet (PLT) (r=0.830,0.692,0.725,0.667,0.388,0.397,0.338,0.476,P＜0.05),L-ficolin levels were positively correlated with DAS28,ESR,CRP,RF,WBC,NEUT,MONO,PLT (r=0.653,0.593,0.630,0.468,0.254,0.302,0.312,0.340,P＜0.05).Conclusion M-ficolin/L-ficolin levels increase in RA patients and correlate with disease activity,which suggests that the complement system may play an important role in the pathogenesis of rheumatoid arthritis.

Objective To observe the clinical efficacy of atorvastatin combined with sodium ferulic acid on renal interstitial fibrosis of diabetic nephropathy(DN).Methods According to the diagnostic and staging criteria of DN,56 patients of DN(Mogensen stage Ⅲ~Ⅳ)who hadn’t received lipid-lowering treatment in the last 2 months and glycemic control were within the standard were randomly divided into control group (n =28 )and experimental group(n=28).Control group were received basic treatment including diabetes diet,decreasing blood glucose level and adjusting blood pressure.Experimental group were added atorvastatin combined with sodium besides the basic treatment above-mentioned.The course is both 4 weeks in two groups.Changes of fasting plasma glucose(FPG),blood urea nitrogen(BUN),serum creatinine(SCr)and 24-hour urine albumin excretion rate (24hUAER),transforming growth factorβ1 (TGF-β1 )before and after the treatment in two groups were observed. Results The differences of FPG,BUN, SCr,24h UAER and TGF-β1 in two groups before and after treatment were statistically significant(P<0.01).Conclusion Atorvastatin combined with sodium ferulate can function cooperatively in the treatment of diabetic nephropathy and can delay renal interstitial fibrosis.

Objective To evaluate the myocardial protective effects of pinacidil combined with pyrrolidine dithiocarbamate (PDTC) against ischemia-reperfusion (I/R) injury to the isolated rabbit hearts and investigate its mechanisms. Methods One-hundred and twelve Japanese long-ear white rabbits of both sexes weighing 1.8-8.2 kg were killed by a knock to the head after heparinization. Their hearts were immediately removed and mounted on Langendorff apparatus and perfused with oxygenated K-H solution at 371 . Of the 112 isolated hearts 96 were randomized into 6 groups with 16 hearts in each group of which 8 hearts underwent 60 min reperfusion and another 8 hearts 120min reperfusion after 40min global myocardial ischemia: the hearts were perfused with K-H solution in group Ⅰ(K); with St Thomas Ⅱ solution in group Ⅱ(S); with pinacidil in group Ⅲ(P); with PDTC + K-H solution in group Ⅳ(PK); with PDTC + St Thomas Ⅱ solution in group Ⅴ(PS) and with PDTC + pinacidil in group Ⅵ(PP) . The rest of the 112 hearts (16 hearts) were perfused with K-H solution for 10 min. Then myocardial tissue was obtained for immuno-histochemical examination (SABC staining) used as normal control value.(1) Time of resumption of heart beat (from the beginning of reperfusion to the resumption of heart beat) was recorded; (2) left ventricular systolic and end-diastolic pressure (LVSP, LVEDP) and + dp/dtmax were monitored; (3) effluent from coronary sinus was collected at 60 min of reperfusion for determination of TNF-? concentration and (4) myocardial tissue was obtained at the end of reperfusion for determination of expression of NF-?B p65 and ICAM-1 in myocardium. Results (1) The heart beat resumption time was significantly shorter in group PP, PS and P than in the other 3 groups (P

Objective To study the relationship between the T helper cells (Th22)/interleukin (IL)-22 and rheumatoid arthritis (RA) with interstitial lung disease (ILD), and to define the clinical significance of Th22 cells for RA. Methods The quantity of Th22 cells in the peripheral blood from 40 patients with RA (20 RA with ILD, 20 RA without ILD) were examined by flow cytometry, the level of IL-22 in the sera was detected by enzyme-linked immunosorbent assay (ELISA). Comparisons between groups were analyzed by t-test, rank sum test, and the correlation of parameters were tested by linear correlation analysis. Results The quantities of CD4+IL-22+ cells (Th22) in RA patients [(0.15 ±0.07)%] were significantly higher than normal controls [(0.09 ±0.05)%] (t=4.097, P<0.01), and IL-22 levels in RA patients [(83 ±7) ng/L] were significantly higher than normal controls [(61±5) ng/L] (t=13.057, P<0.01). The quantities of Th22 cells in RA-ILD patients [(0.18±0.07)%] were significantly higher than RA-NILD patients [(0.13±0.05)%] (t=2.919, P=0.008), and IL-22 levels in RA-ILD patients [(87±6) ng/L] were significantly higher than RA-NILD patients [(80±6)ng/L] (t=3.624, P=0.001). The quantities of Th22 cells were positively correlated with erythrocyte sedimentation rate (ESR), rheumatoid factor(RF) and 1.4 disease activity score (DAS)28 (r=0.336, 0.377, 0.577, P<0.05),and the level of IL-22 were also positively correlated with ESR and DAS28 (r=0.406, 0.576, P<0.05). Conclusion The quantities of Th22 cells and IL-22 level are increased in RA patients, especially in RA-ILD patients. The quantities of Th22 cells and IL-22 level are positively correlated with ESR and DAS28. It may play a certain role in RA especially in RA with ILD.

Objective To compare and analyze the changes of CEA,CA19-9,CA72-4 in different pathologictypes of gastric cancer.Methods 93 patients with gastric cancer were divided into different groups according to the histological types,pathologic types and TNM staging.The levels of CEA,CA19-9,CA72-4 of the patients were measured,then the results were compared and analyzed.Results The level of serum tumor marker of the moderately differentiated group was obviously higher than that of well-differentiated group(P ＜ 0.05),while the level of serum tumor marker of the poorly differentiated group was obviously higher than that of moderately differentiated group (P ＜ 0.05).The differences of levels of CEA,CA19-9,CA72-4 between the well-differentiated group,moderately differentiatedgroup and poorly differentiated group were statistically significant(F =61.433,57.882,125.547,all P ＜ 0.05).The differences of levels of CEA,CA19-9,CA72-4 between patients of TMN Ⅰ stage,TMN Ⅱ stage,TMN Ⅲ stage,TMN Ⅳstage were statistically significant(F =189.624,95.236,80.342,all P ＜ 0.05).The difference of serum tumor marker between different histological groups was not statistically significant (all P ＞ 0.05).Conclusion The concentration of serum CEA,CA19-9,CA72-4 will be higher if the gastric tumor is poorly differentiated or invades deeply,but the concentration of serum tumor has nothing to do with the histological types of the tumor.

Objective To study the application of carcinoembryonic antigen (CEA),carbohydrate antigens19-9 (CA19-9) and carbohydrate antigens72-4 (CA72-4) in the diagnosis of gastric cancer.Methods A total of 78 patients with gastric cancer underwent treatment in our hospital from August,2007 to June,2011 were enrolled as the case group.A total of 78 patients with benign gastric diseases and 78 healthy residents of the same period and district were randomly selected as the benign group and healthy control group,respectively.Measurements of serum CA72-4,CA19-9,CEA of all the cases and control group were performed.And the measurements of the three groups were statistically compared.Results The Serum concentration of CA72-4,CA19-9,CEA were significantly higher than the other two groups [CEA:(27.56 ± 18.36) μg/L vs (2.44 ±0.97) μg/L vs (2.37 ± 1.25) μg/L,F =145.346,P ＜0.0001 ; CA19-9:(99.87 ±80.53) kU/L vs (18.21 ±10.36) kU/Lvs (17.48 ±9.66) kU/L,F=78.503,P＜0.0001; CA72-4:(56.13 ±39.26)kU/L vs (5.77 ±2.95) kU/L vs (3.62 ±2.18) kU/L,F =133.892,P ＜0.0001].And the positive rate of the gastric cancer group were significantly higher than those of the other two groups [CEA:69.23％,10.26％and 7.69％; CA19-9:57.69％,23.07％ and 20.51％; CA72-4:83.33％,10.26％ and 8.97％].The combination of the 3 measurements obtained the highest accuracy of diagnosis,while for single measurement,CA72-4 had the highest accuracy.Conclusion CEA,CA19-9 and CA72-4 have a significant meaning in the early diagnosis of gastric cancer,which has high sensitivity and specificity.The detection rate of the gastric cancer will be improved if the test of CEA,CA19-9 and CA72-4 are combined.

Objective To evaluate the role of nuclear factor-E2 related factor 2 (Nrf2)-antioxidant response element (ARE) pathway in cardio-protection by ischemic or pinacidil postconditioning ( IP,PP) against ischemia-reperfusion (I/R) injury in isolated rat hearts.Methods Fifty-six male SD rats of both sexes weighing 200-250 g were anesthetized with intraperitoneal amobarbital sodium.The isolated rat hearts were perfused in a Langendorff apparatus with Krebs-Hensleit buffer (K-H).Fifty-six isolated rat hearts with I/R injury were randomly divided into 7 groups ( n =8 each):normal control group (group C) ; group I/R; group IP and group PP1-4 postconditioning with 4 different concentrations of pinacidil.After 20 min of equilibration,the perfusion was suspended for 40 min (global ischemia) followed by 60 min of reperfusion in group I/R.In group IP after 40 min of global ischemia,the isolated hearts underwent 6 cycles of 10 s reperfusion and 10 s ischemia followed by 58 min of reperfusion.In group PP1-4 at the end of 40 min of global ischemia,the isolated hearts were perfused with K-H containing pinacidil 5,10,30 and 50μmol/L for 5 min respectively followed by 55 min reperfusion with regular K-H.Left ventricular developed pressure (LVDP) and LVEDP were measured immediately before global ischemia and at the end of 60 min reperfusion.Myocardial specimens were obtained at the end of reperfusion for detection of Nrf2,quinopeoxidoreductase (NQO1),HO-1 and SOD1 mRNA (by RT-PCR) and protein (by Western blot) expression.Results I/R significantly up-regulated Nrf2,NQO1,HO-1 and SODI mRNA and protein expression,decreased LVDP and increased LVEDP in group I/R as compared with group C.IP and 30,50 μmol/L pinacidil postconditioning further significantly increased Nrf2,NQO1,HO-1 and SOD1 mRNA and protein expression and IP,5,10,30,50 μmol/L pinacidil postconditioning significantly increased LVDP and decreased LVEDP as compared with group I/R.Conclusion Ischemic or pinacidil postconditioning can attenuate I/R injury by activating Nrf2-ARE pathway in isolated rat hearts.

Objective To investigate the relationship between the mechanism of ischemic postconditioning-induced activation of nuclear factor-E2 related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway during myocardial ischemia-reperfusion (I/R) and reactive oxygen species (ROS).Methods Healthy male Sprague-Dawley rats, aged 16-20 weeks, weighing 250-300 g, were heparinized and anesthetized with intraperitoneal 1％ pentobarbital sodium 40 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution.Thirty-two isolated rat hearts were randomly divided into 4 groups (n=8 each) using a random number table: control group (group C) , group I/R,ischemic postconditioning group (group IPO) , and N-(2-mercaptopropionyl)-glycine (a ROS scavenger) + IPO group (group M + IPO).After 20 min of equilibration, group C was continuously perfused with K-H solution for 100 min, and the isolated hearts received the drugs via the perfusion system in the other groups.Group I/R was perfused with cardioplegic solution 4 ℃ St.Thomas, and then was subjected to 40 min of ischemia at 32 ℃ followed by 60 min of reperfusion.In group IPO, ischemic postconditioning was induced by 6 cycles of 10 s reperfusion followed by 10 s limb ischemia starting from the onset of reperfusion, and the hearts were then perfused for 58 min.In group M + IPO, the hearts were perfused with K-H solution containing N-(2-mercaptopropionyl)-glycine 2 m mol/L for 3 min starting from the onset of reperfusion,underwent 2 min of ischemic postconditioning, and then was perfused for 55 min.Heart rate (HR), left ventricular developed pressure (LVDP), left ventricular end-diastolic pressure (LVEDP),and positive maximal pressure of left ventricular increase (+dp/dtmax) were recorded at the end of equilibration and of reperfusion.At 5 min of reperfusion and the end of reperfusion, myocardial specimens were obtained from the left ventricle for determination of ROS content by enzyme-linked immunosorbent assay.At the end of reperfusion, myocardial specimens were obtained from the left ventricle for examination of the ultrastructure of myocardial cells and for determination of Nrf2, heme oxygenase-1 (HO-1) , quinone oxidoreductase 1 (NQO1), and superoxide dismutase 1 (SOD1) mRNA and protein expression (by using Western blot and real-time polymerase chain reaction).The damage to myocardial mitochondria was assessed using Flameng scoring.Results Compared with group C, HR, +dp/dtmax and LVDP were significantly decreased, and LVEDP was increased at the end of reperfusion in I/R and M+IPO groups, HR and LVDP were decreased, LVEDP was increased, and no significant changes were found in +dp/dtmax at the end of reperfusion in IPO group, Flameng score was increased in I/R, IPO and M+IPO groups , the ROS content was increased at the end of reperfusion in I/R, IPO and M+IPO groups, and Nrf2, HO-1,NQO1 and SOD1 mRNA and protein expression was down-regulated at the end of reperfusion in I/R, IPO and M+IPO groups.Compared with group I/R, HR, +dp/dtmax and LVDP were significantly increased, and LVEDP and ROS content were decreased at the end of reperfusion, Nrf2, HO-1, NQO1 and SOD1 mRNA and protein expression was up-regulated at the end of reperfusion in IPO and M+IPO groups, Flameng score was decreased in IPO group, there was no significant change in Flameng score in M+IPO group.Compared with group IPO, HR, +dp/dtmax and LVDP were significantly decreased, LVEDP and ROS content were increased at the end of reperfusion, Flameng score was increased, and Nrf2, HO-1, NQO1 and SOD1 mRNA and protein expression was down-regulated in M+IPO group.Conclusion Ischemic postconditioning can regulate ROS level and activate Nrf2-ARE signaling pathway, thus attenuating myocardial I/R injury in rats.

Objective To observe the activation mechanism of Nrf2-ARE pathway in protective effect of ischemia and pinacidil postconditioning on isolated rat hearts.Methods The hearts of adult male Sprague Dawley rats were established ischemia-reperfusion injury model,and devided into six groups(n =8,each group),i.e.Normal group(Group N),ischemiareperfusion group (Group Con,I/R),ischemic postconditioning group (Group IPO),pinacidil postconditioning group (Group P50),N-(2-mercaptopropionyl)-glycine(MPG,2mmol/L) + IPO group(Group M + IPO),MPG + P50 group(Group M + P50).Rat hearts were perfused with Krebs-Henseleit(K-H) buffer for 20 minutes for equilibration.Subsequently,Group N was perfused with K-H buffer for 100 minutes after equilibration,Group Con was perfused with 4℃ ST.Thomas solution to stop the heart beating after equilibration,then the hearts were underwent 40 minutes global ischemia under 32℃,and followed by the K-H solution for 60 minutes.Group IPO after global ischemia period,the hearts were subjected to six 10-seconds cycles of ischemia/reperfusion at the beginning of reperfusion,then were reperfused for 58 minutes.Group P50 after global ischemia,rat hearts were perfused with K-H buffer containing pinacidil(50.μmol/L) for 2 minutes before reperfusion.Group M + IPO after global ischemia,the hearts were subjected to perfuse with K-H buffer containing MPG(2 mmol/L) for 3 minutes,and then underwent six 10-seconds cycles of ischemia/reperfusion before reperfusion.Group M + P50 after global ischemia,the hearts were perfused with K-H buffer containing MPG(2 mmol/L) for 3 minutes,and then subjected to perfuse with K-H buffer containing pinacidil(50 μmol/L) for 2 minutes before reperfusion.Cardiac function indexes(such as HR,LVDP,LVEDP,and the Max dp/dt) at the end point of equilibration and repeffusion were observed and recorded.The ultrastructure of myocardial tissue was observed by electron microscopy and the mitochondrial Flameng score was calculated.RT-PCR and western-blot were applied to detect the gene transcription and protein expression of HO-1,NQO1,SOD1,and Nrf2 in left ventricular myocardial tissue after reperfusion.Results The HR,LVDP and + dp/dtmax at the end of reperfusion:the cardiac function indexes are lower among each group compared with group N,group 1PO and group P50 are better than group Con (P ＜ 0.05).Compared with group IPO,there is no significant difference in group group P50,but group M + IPO is obviously decreased(P ＜ 0.05).Compared with group P50,group M + P50 index is decreased significantly(P ＜ 0.05).The LVEDP at the end of reperfusion is lower than that among each group as compared with group Con,which is significantly increased in group Con (P ＜ 0.05).Compared with group IPO,there is no significant difference in group P50,but group M + IPO is significantly increased(P ＜ 0.05).Compared with group P50,the group M + P50 is obviously decreased(P ＜ 0.05).The ultrastructure of myocardial tissue in group N is mostly normal,group Con presence serious damage.The ultrastructure damage of myocardial tissue is improved in group IPO and group P50 as compared with that in group Con,while group M + IPO is more serious than group IPO,group M + P50 is more serious group P50.The mitochondrial Flameng score is higher among each group as compared with group N (P ＜ 0.05),the score is lower in group IPO and group P50 as compared with group Con and corresponding nonblocking group (M + IPO,M + P50,P ＜0.05).The mRNA and the protein expressions of HO-1,NQO1,SOD1 and Nrf2 among each group are lower as compared with group N(P ＜0.05).Compared with those in group Con,the mRNA and the protein expressions in group IPO and group P50 are obviously increased(P ＜ 0.05),group IPO and group P50 are higher than those in group adding active oxygen scavenger(MPG) (P ＜ 0.05).Conclusion Ischemic postconditioning and pinacidil postconditioning have protective effect of myocardial tissue from ischemia reperfusion injury,while improve the cardiac function index.The cardiac protective effect of Ischemic and Pinacidil postconditioning methods may be involved the ROS in early reperfusion,which activate the Nrf2-ARE pathway,and up-regulate the expression downstream antioxidant protein and phase Ⅱ detoxifying enzyme,ultimately improve the cardiac function index during the reperfusion period.