OBJECTIVE:To explore the ways of preventing errors in the process of drug treatment.METHODS:Related overseas literatures were consulted;causes of errors in the drug treatment were analyzed,which combined with prac-tice.RESULTS&CONCLUSIONS:Errors can be found effectively and reduced by establishing independent checking and error reporting system.

ObjectiveTo evaluate the therapeutic effect of vitreo-retinal surgery on oclular siderosis. MethodsThe clinical data of 22 patinets (22 eyes) with ocular siderosis due to the magnetic foreign body at intraocular postsegment were retrospectively analyzed. The patients aged from 6 to 54 years (average 40 years), including 21 males and 1 femal. The duration of the magnetic foreign body remained in the eye lasted for 1 month to 20 years. The preoperative best corrected visual acuity (BCVA) was <0.01 in 15 eyes, 0. 01-0. 15 in 5 eyes and 0.1-0.2 in 2 eyes. There was Intra-vitreous foreign body in 18 eyes and ocular wall embedded foreign body in 4 eyes; intraocular foreign body (IOFB) combined with cataract in 18 eyes; combined with retinal detachment in 3 eyes; scleral buckling combined with silicon oil filled in 12 eyes and C3F8 filled in 7 eyes.Cataract extraction was performed in 12 eyes, and 2 eyes underwent filtrating surgery. ResultsThe IOFB was successfully removed by one-off surgery in 22 eyes. BCVA increased in 20 eyes (90.9%) and kept unchanged in 2 eyes (9. 1%), including<0.1 in 7 eyes, 0. 1-0.4 in 8 eyes, and 0.5-1.0 in 7 eyes. Operative complications involved retinal holes with retinal detachment in 2 eyes and vitreous haemorrhage secondary to enlarge sclera incision in 2 eyes.Postoperative complications included secondary cataract in 4 eyes, retinal detachment due to silicon oil removal 3 months after submacular removal of foreign body in 1 eye, and retinal detachment 7 days after C3F8 filling in 1 eye; the latter two eyes had reattached retina after another silicon oil filling. At the end of the follow-up period, retina reattached in 22 eyes. ConclusionAdvanced modern vireo-retinal operation is ffective on oclular siderosis, which can avoid the release of Fe+ and improve the patients' visual function.

BACKGROUND:Mesenchymal stem cel s as potential seeded cel s have been widely used in tissue engineering and clinic therapy;thus, the precise, safe, effective isolation of mesenchymal stem cel s is the particular important premise to build culture system.
OBJECTIVE:To review the methods of isolating mesenchymal stem cel s and to compare the merit and demerit of different methods, thereby providing theoretical basis for safe and high-effective isolation of mesenchymal stem cel s.
METHODS:A computer-based online research of CNKI and PubMed databases was performed to col ect articles, which included reviews, clinical trials and experiments, published between 1965 and 2014 with the key words of“mesenchymal stem cel s (MSCs), isolation methods”in Chinese and English. A total of 52 articles were included according inclusion and exclusion criteria
RESULTS AND CONCLUSION:(1) The whole bone marrow culture method can derive a mass of mesenchymal stem cel s, which need to be purified. (2) The density gradient centrifugation method which uses the media with the density of 1.073 g/mL can be used to harvest more purified cel s. (3) The tissue digestion method is suitable for digestion and isolation of adipose tissue and umbilical cord tissue. Type II col agenase digestion is better, but they are both limited by a high demand for operative techniques. (4) Immunomagnetic bead separation is appropriate to study the biological characteristics of a kind of subpopulation of mesenchymal stem cel s which express special surface markers. (5) The combination method is also an optimal way. (6) Some new methods limited by few dates require further studies.

Objective To successfully get the full PCR alleles of the Insertion/Deletion marker rs10644346 in which a SNP-binding exists at the 3'end region of the primer.Methods Based on the AS-PCR,a common forward primer and two reverse primers with allele-specific oligonucleotides at the last second 3'end instead of the terminus were tentatively designed for typing 150 unrelated individuals and 10 father-mother--child trios from Htnan province in South-central China.Simultaneously,9 samples were typed with all the above three primers (the two primer sets which consist of the common forward primer and one of the reverse primers).Results PCR amplicons were well detected in the 150 unrelated individuals after being typed with the three primers,and the amplified fragments of parental and filial generations among 10 father-mother-child trios conformed to Mendel's principles.Allele missing was found in the two-primer group.Conclusion The primers designed by locating the specific nucleotide at the last second 3'end rather than terminal position were demonstrated also effective in getting specific alleles if perfect mismatch and PCR conditions are guaranteed,and the design strategy can provide an optional reference to rescue markers of SNP-binding primers for forensic practice.

BACKGROUND:In previous studies, it is satisfied to sorting bone marrow mesenchymal stem cels based on natural sedimentation combined with low permeation. Then, based on particle hydromechanics theory, the settling velocity of bone marrow mesenchymal stem cels in culture liquid is calculated. A simple and easy method of separation, purification and proliferation of bone marrow mesenchymal stem cels is established by natural sedimentation velocity. OBJECTIVE:To explore the feasibility of sorting bone marrow mesenchymal stem cels by natural sedimentation velocity method. METHODS:The density interval of rabbit bone marrow mesenchymal stemcels (ρ1) was determined by density gradient centrifugation method. The diameter of rabbit bone marrow mesenchymal stem cels (d) was measured by scanning electron microscope. The density of culture liquid (ρ2) was measured by liquid density meter. The viscosity of the culture liquid (μ) was measured by viscosity meter. The settling velocity of bone marrow mesenchymal stem cels (Vt) was derived from the above four numerical values with the appropriate formula. Bone marrow mesenchymal stem cels were sorted from bone marrow tissue by natural sedimentation velocity method. Cel proliferation, purity and differentiation were observed. Meanwhile, the primary culture time of three different cel sorting methods was recorded; the colony formation rate of rabbit bone marrow mesenchymal stem cels was determined. RESULTS AND CONCLUSION:(1) The diameter of rabbit bone marrow mesenchymal stem cels was (20.37±4.58) μm, and the setting velocity of rabbit bone marrow mesenchymal stem cels in the culture liquid was 50-55 mm/h. (2) Bone marrow mesenchymal stem cels could be successfuly isolated from bone marrow tissue of rabbits by the natural sedimentation velocity method, which could be induced into osteoblasts and adipocytes. (3) The natural sedimentation velocity group cost less time than the other two density gradient centrifugation groups in the primary culture stage. The colony formation rate of the natural sedimentation velocity group was higher than that of the other two groups. (4) Natural sedimentation velocity method did not impose any intervention measures for sorting cels, which maybe maximaly maintain cel viability and biological characteristics. The whole separation process was simple and safe, which may have little damage to the cels.

Objective To investigate the dynamic changes of osteopontin (OPN) in the guinea pig model of cholesterol gallstone disease.Methods Forty-four guinea pigs were randomly assigned to cholesterol gallstone group and control group.The animals were sequentially killed on Day 7,14,28,42,56 and 70 after operation.The expressions of OPN mRNA in gallbladder and liver tissues were detected by real-time PCR.The changes of OPN,mucin,α1-acid glycoprotein (AAP) and apolipoprotein A1 (APOA1) in bile and blood plasma were detected by ELISA kits.Results The expression of OPN mRNA in gallbladder and liver tissues increased gradually and reached the peak in the 6th week,and then decreased.The increased expression of OPN in bile in the gallstone group started from the 1st week and reached the peak in the 6th week (P ＜ 0.05),which gradually decreased to the baseline in the 10th week (P ＞ 0.05).The expressions of OPN in bile and blood demonstrated similar trends,while the peak time in blood samples was much earlier (4th week).The changes of APOA1 in bile and blood were similar to OPN,although there was no advanced peak value in blood.The levels of mucin and AAP in bile and blood increased after operation,and were kept at high level throughout the study.Conclusions OPN is involved in the whole process of cholesterol gallstone formation,which may be associated with other nucleation-active factors.

Objective To investigate the role of osteopontin (OPN) in the pathogenesis ot cholesterol gallstone formation in bile.Methods The nucleation time of OPN in model bile and human gallbladder bile was studied by the nucleation time assay,the effect of OPN on cholesterol crystal growth in model bile was examined by the cholesterol crystal growth assay.The effect of OPN on vesicle was detected by the transmission electron microscopy in model bile and gallbladder bile; then the content of OPN and calcium were detected via the commercial kits in human bile.Results Osteopontin prolonged nucleation time in a dose dependent manner in model bile and human bile,and this effect was correlated with calcium.Compared with control group,the nucleation times were prolonged by 1.50and 1.93 times in lithogenic bile at the concentration of osteopontin 50 μg/ml and 100 μg/ml (P＜0.01),respectively. Nucleation time were prolonged by 1.17 and 1.33 times in normal bile (P＜0.01) and by 1.29 and 1.48 times in model bile (P＜0.01),respectively.The rate of cholesterol crystals growth was not influenced by calcium ions,but inhibited by osteopontin in a dose dependent manner in the model bile.Furthermore,the formation,aggregation and fusion of vesicles were delayed by osteopontin in bile samples as indicated by the transmission electron microscopy.The concentration of osteopontin [(0.53± 0.08) mg/ml vs. (0.65 ± 0.14) mg/ml,P＜0.05] and the calcium ions [ (0.71 ± 0.17) mmol/L vs. ( 0.84 ± 0.08 ) mmol/L,P ＜ 0.05 ] were lower in lithogenic bile than in control.Conclusions Osteopontin can inhibit the cholesterol gallstone formation in model and human gallbladder bile as the anti nucleating factor.

Objective To observe the effect of combined treatment with rhTRAIL(recombinant human TNF-related apoptosis inducing ligand) and selective Cox-2 inhibitor Celecoxib on gallbladder carcinoma in vitro and to explore the possible mechanism of the effect. Methods Western blot analysis was used to detect the expression of c-FLIP and death receptors after treatment by Celecoxib. Apoptosis of gallbladder cell line SGC-996 after the combined treatment with Celecoxib and rhTRAIL was detected in three ways: (1) phase microscopy of the cells, (2) detection of effector caspase-3 and caspase-7 activity, and (3) determination of the proportion of apoptotic cells labeled by Annexin V-PI flow cytometric analysis using CELLQUEST software. Results Celecoxib down-regulated the expression of c-FLIPs and up-regulated the expression of DR5 in a dose- and time-dependent mode on cell line SGC-996. Apoptotic levels in the combined treatment group in cell line SGC-996 were significantly higher than those in the single drug treatment group and control group. Conclusion Celecoxib markedly sensitized rhTRAIL-induced apoptosis through the down-regulation of c-FLIPs and up-regulation of DR5 in gallbladder carcinoma cell line SGC-996.

Objective To investigate the role of osteopontin (OPN) in cholesterol gallstone formation.MethodsGallbladder bile was obtained from patients with cholelithiasis (n=36,the experimental group) and from donors of liver transplantation (n=19,the control group).OPN,calcium ion and lipid were analysed quantitively.The nucleating role of OPN in bile was evaluated using nucleating time (NT) approach.ResultsOPN inhibited cholesterol nucleation in a dose dependent manner.OPN (50 μg/ml and 100 μg/ml) prolonged NT by 48.90％ (91.51％) and 17.07％ (32.93％) in lithogenic and control bile,respectively.OPN (100 μg/ml) also inhibited the nucleating effect induced by calcium ion.Furthermore,a combination of OPN (50 μg/ml) and calcium prolonged NT by 75.78％ and 33.96％ in lithogenic and control bile,respectively.A combination of OPN (100 μg/ml) and calcium prolonged NT by 125.9％ and 62.26％ in the 2 groups.The contents of osteopontin and calcium were significantly lower in lithogenic bile than control bile (P＜0.05).On the other hand,the cholesterol saturation index and the contents of cholesterol,phospholipid and bile acid were significantly higher (P＜0.05).ConclusionsOPN inhibited cholesterol gallstone formation.It may be involved in the pathogenesis of cholelithiasis.

AIM To research effects of sodium ferulate(SF) on transforming growth factor ?_1 inhibited hepatocyte growth. METHODS Human cultured hepatocytes(L02 cells) served as control group ,TGF?_1(5 ?g?L -1 ) was used to inhibited L02 cells to construct the cell model. MTT methods were used to study the growth inhibitory effect of TGF?_1 on L02 cells. DNA synthesis was analyzed by measuring 3H-TdR incorporation. The effects of SF on the changes of cell cycle were analyzed by flow cytometry. Generation of intracellular ROS was determined by the emission of fluorescence in L02 cells proloaded with DCFH-DA fluoresin and then treated them with TGF?_1 or SF. After 4h,control, TGF?_1 and TGF?_1+SF-treated cells were analyzed by flow cytometry. RESULTS ①Incubated in the presence of TGF?_1 for 24 h, 67.93% of L02 cells survived (P