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 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.

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 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 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 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.

Background Accurate measurement of corneal thickness is very important during the pre-and post-operative management of corneal surgical procedures,especially laser-assisted in-situ keratomileusis (LASIK),which is the most popular approach to the correction of refractive errors currently.This may be particularly important for the patients who have undergone previous laser refractive surgery with suboptimal outcomes and are being considered for an enhancement procedure.Objective This study was to compare the measuring outcomes of corneal thickness by slit-scanning pachymetry,non-contact specular microscope,anterior segment optical coherence tomography (AS-OCT)and ultrasound pachymetry,with a focus on central and midperipheral (from the central 3.0 mm) region of cornea in post-LASIK eyes.Methods Sixty-four right eyes of 64 patients who received LASIK were collected in Henan Eye Institute,Henan Eye Hospital from March to June 2011 with the equivalent spherical diopter of (-4.75±2.38)D and horizontal corneal diameter of (11.36±0.32)mm.Central corneal thickness was measured on each eye by using non-contact specular microscope (Topcon SP-3000P),slit-scanning pachymetry (Orbscan Ⅱ),AS-OCT and A-type ultrasound pachymetry,respectively,and the paracentral corneal thickness including 12:00,2:00,6:00 and 10:00 meridian was measured using Orbscan Ⅱ,non-contact specular microscope and AS-OCT.The measuring values and the agreement from different instruments were compared and evaluated.Results The mean central corneal thickness was (467.12±31.10)tμm for AS-OCT,(466.67±30.99)μm for ultrasound pachymetry,(441.84 ± 33.65) μm for specular microscopy and (422.51 ± 44.09) μm for Orbsan Ⅱ,respectively,showing a significant difference among the four methods (F =23.730,P =0.000).The central thickness value of the A-type ultrasound pachymetry was significantly higher than that of Orbsan Ⅱ or non-contact specular microscope (q =6.940,6.720,both at P =0.000).Compared with Orbscan Ⅱ,the measuring values of non-contact specular microscope and AS-OCT were significantly higher (q =-5.54,6.940,both at P =0.000),and the measuring value of AS-OCT was significantly higher that of non-contact specular microscope (q =6.800,P =0.000).The lowest difference value (25.3 μm)and the best agreement was found between the ultrasound pachymetry and AS-OCT.The paracentral corneal thickness values in 12:00,2:00,10:00,6:00 meridians were highest for Orbsan Ⅱ and the next for AS-OCT,and non-contact specular microscope had the lowest values,with significantly differences among them (F =5.020,22.950,67.890,18.850,all at P ＜ 0.01).Conclusions The corneal thickness values vary with the different instruments.Orbsan Ⅱ underestimates the central corneal thickness and overestimates the midperipheral corneal thickness,and non-contact specular microscope underestimates both the central and midperipheral corneal thickness.The measuring outcome from AS-OCT has a good agreement with ultrasound pachymetry and therefore they can be used interchangeably.

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 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.