MicroRNAs (miRNAs)are involved in the pathological and physiological process of life activities,and some of them exist stably and abundantly in serum,plasma,and other body fluids.Previous studies have indicated that there are significant differences in the expres-sion patterns of circulating miRNAs between patients with viral hepatitis,liver fibrosis,liver cirrhosis,hepatocellular carcinoma,drug-in-duced hepatitis,and other hepatobiliary diseases at different disease stages.Among these circulating miRNAs,some can act as novel nonin-vasive biomarkers for the early diagnosis,disease progression,and prognostic evaluation of related diseases.The latest research advances in circulating miRNAs for the diagnosis of hepatobiliary diseases are reviewed.

[Objective] To observe the clinical therapeutic effect of XBJ oxygen inhalation on chronic pulmonary-cardiac disease in acute period.[Method] Randomly divide 150 cases into blank control group(1),vein administration group(2),oxygen inhalation group(3),observe their clinical effect,lung function,blood-gas analysis,cardiac function grade and other indexes.[Result] Group 3 could improve patients lung function,blood-gas analysis,cardiac function grade and other indexes,which had marked advantages compared with other 2 groups.

OBJECTIVE: To analyze the occupational hazardous factors in hospital pharmacy so as to enhance pharmacist’s awareness of self-protection against such hazards. METHODS: The occupational hazardous factors including physical factors, chemical factors and psychological factors etc were analyzed and the countermeasures were discussed. RESULTS & CONCLUSIONS: Pharmacists should be cautious to the occupational hazards, strengthen traning and education, raise awareness of self-protection to guard against the occupational hazard event.

OBJECTIVE:To provide references for the evaluation of drug suppliers in hospital so as to promote the suppliers to enhance their service level. METHODS: The drug suppliers were evaluated comprehensively with on-time delivery percentage, drug price and post-sale service as evaluation criteria. RESULTS & CONCLUSIONS: The evaluation of drug suppliers can not only raise the service level of the suppliers, but also facilitate a scientific and standardized management of drug supply. However, great importance should be attached to the impartiality and fairness in the evaluation.

Objective To study the effect of Chinese herb medicine compound on general state and bone-muscle system in simulated weightlessness rats, and to observe the synergistic action of other ingredients in the compound on calcium. Methods Thirty Wistar rats were divided into 3 groups:control group, tail suspend group, tail suspend and medicine group which were given Chinese herb medicine compound by intragastric administration. After 3 weeks simulated weightlessness, body weight (BW), muscle weight (MW) and index (MI) of posterior limb, bone length (BL), wet weight (BWW), index (BI), dry weight (BDW), content of organic (ORG) and inorganic (INO) substance, bone mineral density (BMD), and mechanical properties (MEC) of femur were observed. Results At the middle-later stage of the experiment, BW of tail suspend group decreased significantly (P

Objective To observe the efficacy of melbine combined with clomifene citrate in treatment of patients with polycystic ovarian syndrome ( PCOS) .Methods 60 patients aged 20 to 39 with PCOS were randomly divided into observation group and control group, 30 patients in each group.In the menstrual cycle of 3~7 d, the control group received clomifene citrate orally (50 mg per time, qd), and the observation group received melbine (2.5 mg per time, qd)combined with clomifene citrate (50 mg per time, qd), with a course of 5 days.Results The number of mature follicles and score of cervical mucus in obsercation group was significant higher than that in control group, respectively(P<0.05).The rate of follicular development and ovulation rate of single follicle in obsercation group was significant higher than that in control group, respectively(P<0.05), but there were no significant differences of pregnancy rate, first trimester abortion rate and periodic ovulation rate between two groups.The rate of endometrial thickness<7 mm patients in observation group was significantly lower and positive rate of three line was higher than that in control group, respectively ( P<0.05).The follicle stimulating hormone level in observation group was significantly lower than that in control group(P<0.05), but there was no significant difference of luteinizing hormone between two groups.Conclusion Melbine combined with clomifene citrate has a good effect on mature follicles number and endometrial thickness in anovulatory infertility women caused by polycystic ovary syndrome.

Objective The fluorescence-activated cell sorting (FACS) technique is a method for the identification and isolation of different cell populations.At present,the special surface marker for limbal stem cells has been not found yet.This study aimed to investigate the application of FACS technique in the research of rabbit limbal stem cells.MethodsCorneal limbal tissue was obtained from New Zealand white rabbits and cultured using the explant culture method in SHEM.Side population cells (SP cells) and non-SP cells were sorted from cultured rabbit limbal epithelium cells by FACS at a excitation wavelength 350 nm,and acquistion length 450 nm (blue light) and 675 nm (red light).The SP cells and non-SP cells were identified by detecting the expression of ABCG2 and K3/K12.The colony-forming efficiency of SP cells and non-SP cells were evaluated by the observation of cellular vitality with trypan blue staining.The percentage of colony formation was calculated as the colony number in various group/200×100%.ResultsIn 48-72 hours after primary culture,limbal epithelial cells migrated from the cultured tissue mass to form the mambrane-like structure and achieved 70%-80% confluence.The cells showed round,polygon and flattened shape.The proportion of SP cells in cultured limbal epithelial cells was 0.22%±0.09% with a colony-forming efficiency of 5.52±0.45% in SP cells and 0.78%±0.73% in non-SP cells,with a statistically significant difference between the two populations (t＝2.17,P＜0.01).After verapamil,an inhibitor of the expression of the ABCG2 protein,was added into the medium,the proportion of SP cells in the cultured limbal epithelial cells declined to 0.04%±0.006%.The SP cells presented a positive immunoresponse for ABCG2 and absence of immunoresponse for K3/K12,but a contradictory staining result was found in non-SP cells.ConclusionFACS can be applied in the research of limbal stem cells.

BACKGROUND:Compared with traditional autogenous bone graft, composite, as the carrier of seed cells, possesses advantages of fewer traumas and no limitation of donor site in repairing bone defect.OBJECTIVE: To observe the ability of the composite of β-tricalcium phosphate artificial bone-hyaluronic acid-type I collagen (β-TCP/HA/COL-I), as induced bone marrow stromal cell (MSC) carrier, to repair rabbit radial defect, and the feasibility with the composite as bone substitute material.DESIGN: A randomized and controlled trial.SETTING: Department of Orthopaedics, Renmin HospitaL, Wuhan University.MATERIALS; The study was conducted in the Department of Orthopaedics Renmin Hospital, Wuhan University between September 2003 and July 2004. Thirty-one New Zealand big white rabbits, aged 6 months, with body mass of 1.5 to 2.0 kg were enrolled in this study. The rabbits were randomized into control group (n =4) and experimental group (n =27).METHODS : ①In vitro induction and culture of MSCs was performed on 31 white rabbits, and the alkaline phosphatase (ALP) positive ratio of induced MSCs was observed. The structure of β-TCP/HA/COL-I was observed under scanning electron microscope. ② A 2 cm radial defect was created through operation. Eight weeks later, composites of β-TCP/HA/COL-I/MSCs were implanted into one side of rabbits in the experimental group, and autogenous bone was implanted into the other side. Rabbits in the control group were untouched. ③All the animals in the experimental group were randomly sacrificed at postoperative 4,8 and 12 weeks, 6 rabbits at 4 and 8 weeks, 15 at 12 weeks; Animals in the control group were sacrificed at 12 weeks. Gross observation, X-ray photographing, haematoxylin-eosin (HE) dyeing, and assessment of inorganic ingredient were performed. Osteogenic area and biomechanical tests were performed at 12weeks. Repairing effects on bone defect in each group were compared.MAIN OUTCOME MEASURES: The ALP positive ratio of induced MSCs; The structure of composite ofβ-TCP/HA/COL-I;Gross observation; X-ray photographing; HE dyeing and assessment of inorganic ingredient; Osteogenic area and biomechanical tests.RESULTS: All the 31 rabbits entered the stage of result analysis. ① The ALP positive ratio of cells reached 75% after induction and culture. ② Scanning electron microscope observation showed that 3 kinds of materials with abundant cellular structure distributed evenly. ③ The osteogenic area at 12 weeks was (72.5±3.6)% and (76.7±6.2)% in the experimental group and autogenous group, respectively (P ＞ 0.05). ④The maximum bending moment was (521.0±61.1) and (554.3±53.3)N·mm in the experimental group and control group, respectively; The maximum displacement at point of application of force was (0.816±0.071)and (0.870±0.103)respectively, without significant difference (P ＞ 0.05). ⑤Inorganic ingredient in the composite was 75%, 57% and 42% at 4,8 and 12 weeks respectively, suggesting that the inorganic component in the material was gradually decomposed with the elongation of time. ⑥Results of gross observation,X-ray photographing, histopathological examination, biomechanical test showed that with the elongation of time, composite of β-TCP/HA/COL-I/MSCs could repair bone defect in the experimental group, while bone defect in the control group had not been repaired.CONCLUSTON: Composite of β-TCP/HA/COL-I /MSCs possesses the same effect on repairing bone defect as autogenous bone, so it may be used as autogenous bone graft substitute.

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.