BackgroundMüller cells has been recognized as being vital in both healthy and diseased retina.Recently,these cells even have been identified to be the source of retinal progenitor cells.In order to study the possible function of the retinal Müller cells,it is important to establish a practical procedure to obtain the purified cells. ObjectiveThis study was to simplify the procedure of primary culture and purification of retinal Müller cells in vitro. MethodsEyeballs of SPF newborn SD rats were enucleated and retinas were dissected free after soaking the eyeballs overnight in DMEM/F12 medium in room temperature.Then the retinas were mechanically dissociated into micro aggregates and cultured in DMEM/F12 medium containing 10％ FBS for 8-10 days.The floating retinal aggregates and debris were removed and the medium was changed in 2-3 days interval to get more purified flat cell population.Cultured cells were passaged after confluent.Immunofluorescence staining was used to detect the response to glutamine synthetase (GS) and Vimentin for the identification of the cultured Müller cells,and flow cytometry (FCM) was used to estimate the purity of the cells. Results Cultured Müller cells had large cellular body and richer cytoplasm.More than 95％ of the cells showed the positive response for GS with the brown staining in cytoplasm and cellular nuclei,and the positive stainiug also was seen for Vimentin in cytoplasm.FCM showed that 99.7％ of the cells were GS positive after 3 passages.Conclusions Modified procedure in this experiment is a simple and practical method for culturing retinal Müller cells.

Retina is subjected to many acquired and inherited neuronal degenerative diseases such as agerelated macular degeneration (AMD), diabetic retinopathy (DR) and retinitis pigmentosa (RP). All of these diseases are associated with the progressive damage and loss of photoreceptors, which is causing visual impairment and irreversible blindness. Stem-cell therapy is being widely considered as a promising treatment of these incurable retina diseases. However, in mammals including humans, there seems to be little or no recovery of lost cells. By contrast, nonmammalian vertebrates, such as amphibians and fish, have robust regenerative responses to injury, which can lead to the near complete restoration of the neurons lost through injury. Nevertheless ,over the past several years, studies have investigated that stem cells do exist in the adult mammalian eyes, and at least some types of neurons can be regenerated in the mammalian retina by stimulating with growth factors or transcription factors. These recent results suggest that some part of the regenerative program occurring in lower vertebrates remains in the mammalian retina.Here, the origin of various of adult retinal stem cells for the self-renewal and proliferation. and the relevant influencing factors were summarized.

Coupled magnetic nanoparticles in the microcapsule structure, such as magnetic microcapsules, can be delivered in specific organism or tissues under magnetic field exposure. Thus, the microcapsules can achieve active targeting functions by manipulat-ing the magnetic field. Based on the magnetic microcapsules, the antitumor drugs can also be loaded to realize magnetic response, which gives microcapsules sustained and controlled release advantages. To date, the drug microcapsules carrying magnetic nanoparti-cles have become promising novel delivery carriers for the treatment of tumor diseases. This paper mainly reviews the method of prepa-ration of the magnetic nanoparticle-coupled microcapsules, including liposomes, polyelectrolyte microcapsules, and polymer micro-spheres. The basic research progress of these microcapsules as anticancer drug carriers for the tumor therapy was also reviewed.

OBJECTIVETo evaluated the safety and effect of the 120-W GreenLight HPS laser photoselective vaporization of the prostate (PVP) in different BPH populations.

METHODSThis study included 174 BPH patients treated by PVP using 120-W Green-Light HPS laser. According to the prostate volume (PV) ( < 80 or ≥ 80 ml), history of 5-alpha reductase inhibitor (5-ARI) medication, and history of acute urinary retention (AUR), we divided the patients into a PV < 80 ml, a PV ≥ 80 ml, a 5-ARI, a no 5-ARI; an AUR, and a no AUR group. We collected the baseline, perioperative, and follow-up data about the patients, and compared them among different groups.

RESULTSThe patients were aged 69.4 ± 7.7 years, of whom PVP was successfully performed for 136 and PVP was intraoperatively converted to transurethral resection of the prostate (TURP) in the other 38, with a mean operation time of (49.4 ± 16.3) min. The preoperative PV averaged (67.9 ± 29.8) ml. There was no intraoperative blood transfusion, transurethral resection syndrome, or capsule perforation. Bladder neck contracture occurred in 2 cases and urethral stricture developed in another 2 postoperatively. All the patients showed a significant improvement in the prostatic function parameters and no significant differences were observed between the PV < 80 ml and PV ≥ 80 ml, 5-ARI and no 5-ARI, or AUR and no AUR groups.

CONCLUSIONPVP with 120-W GreenLight HPS laser is safe and efficient for the treatment of BPH and the its effect is not influenced by the prostate volume, history of 5-ARI medication, or history of AUR. However, preoperative urinary catheterization may increase the difficulty of surgery and the risk of conversion to TURP.

5-alpha Reductase Inhibitors ; therapeutic use ; Aged ; Humans ; Laser Therapy ; adverse effects ; methods ; Male ; Operative Time ; Organ Size ; Prostate ; pathology ; surgery ; Prostatic Hyperplasia ; pathology ; surgery ; Transurethral Resection of Prostate ; Treatment Outcome ; Urethral Stricture ; etiology ; Urinary Bladder ; Urinary Retention

Objective To understand the epidemic situation of Hepatitis B of the children and the situation of Hepatitis B vaccine injection in Maanshan city so as to provide the basis for the development of Hepatitis B vaccination strategy or adjustment in all of the people. Methods Two-stage sampling method was designed to survey 978 children under the age of 15 in 8 villages, with 3～5 ml of venous blood brawn and HBVM detected by ELISA. Results HBsAg positive rate of the children under the age of 15 was 10.12% in 1991 and, compared with the present rate at 0.92%, there was a significant difference. The protection rate of Hepatitis B vaccination was 90.91%. HBsAb positive rate of the children under the age of 15 was 40.49%, and the distribution was balanced among regional, urban and rural areas. With the in crease in age, the trend of Hepatitis B antibody levels declined, but the 1～5 age group had lower peak. The survey rate of Hepatitis B vaccine injection of the children under the age of 15 was 91.82%, and the report rate was 87.89%. The regional difference for them was significant. With the in crease in age, the rate of Hepatitis B vaccine injection declined for the survey and the report in all of the children under the age of 15. Conclusions The effect of Hepatitis B Control for 16 years is very significant in the city. It is necessary to administer charge-free Hepatitis B vaccination for the newborns and to strengthen the Hepatitis B vaccination in flowing children.

Objective:To generate monoclonal antibodies against ciprofloxacin (CIP) and to analyze its immunological traits for establishing a determination method for ciprofloxacin residues in food tissues.Methods:BALB/c mice were immunized by the conjugation of CIP-BSA successfully.The splenic cells of BALB/c mice and the oncocyte were fused and screened in HAT culture medium.Cell strains of 1C9,3F6,6H2,6A7,6G11 and 8F5 were cloned,which could secret the monoclonal antibody (Mab) for ciprofloxacin.Results:By ELISA method,the haplotype of Mabs were identified.The antibody secreted by 1C9,3F6 and 6A7 were classified to the IgG2a.Those of 6H2 and 8F5 were classified to the IgG1 while 6G11 to IgG3.The results of SDS-PAGE showed that the Mab protein was made up of the weight chain and the light chain whose molecular weight were 50 kD and 25 kD,respectively.All of those Mabs had fine specificity and sensibility.The indirect ELISA titer of 1C9 was 1:6.4?102 in supernatant and 1:5.6?105 in ascites.The affinity of 1C9 was 2.85?109 L/mol and the value of IC50 reached to 245.86 ng/ml.The protein secreted by 1C9 was screened for establishing the ELISA method and the detective limitation reached to 45.25 ng/ml.There were no cross reactions to Ofloxacin,Difloxacin,Sarafloxacin,Malachite green,Chloramphenicol and Furacilin while the rate of cross reaction to Enrofloxacin could reach to 84.6%.Conclusion:The Mabs are sensitive and specific,which are suitable to be applied in establishing immunoassay of CIP residues.

Objective To study the lymphoproliferative responses to HHV-6(Human Herpesvirus-6,HHV-6)among patients with oral squamous cell carcinoma,and to discuss the role of HHV-6 in the pathogenesis of oral squamous cell carcinoma.Methods IgG antibody to HHV-6 in plasma was identified by indirect immunofluorescence(IIF).Immuno-magnetic beads were used to prepare CD4+ T cells and CD8+ T cells.The proliferation of CD4+ T cells,CD8+ T cells and PBMCs stimulated with HHV-6 or anti-CD3 antibodies was measured by 3H-thymidine uptake.The percentage of CD4+ C25+ Treg cells(Regulatory T cell,Treg)within the peripheral blood CD4+ T cell compartment was analyzed by flow cytometry.Results Significantly higher proportion of patients with oral squamous cell carcinoma had IgG antibody to HHV-6(8/8) in plasma as compared with those in control subjects(12/20);The proliferative responses of PBMCs and CD4+ T cells from patients with oral squamous cell carcinoma to HHV-6 were significantly decreased as compared to those from HHV-6-infected and healthy individuals(P

Objective To investigate the microsatellite instability(MSI) and monomorphism at microsatellite DNA BAT 26 locus in esophageal mucosa. Methods Genomic DNA extracted from tissues was amplified by PCR. PCR products were run on 9% denaturing polyacrylamidegel and stained with silver. Then MSI and monomorphism of microsatellite at BAT 26 locus in normal tissues at the incised edge and esophageal cancer tissues in 45 cases of esophageal caner were analyzed. Results All normal specimens showed no change in the length of MS DNA at BAT 26 locus. MSI was detected in 3 out of 45(6 7%) cases of esophageal cancer. Conclusion BAT 26 has complete monomorphism in genome of normal esophageal tissues, but is not sensitive to the identification of MSI.

Objective To detect the state of microsatellite instability (MSI) and investigate the relationship between MSI and clinical pathological parameters in esophageal cancer. Methods MSI was analyzed by PCR SSCP method. Results MSI was detected (22 22%) in 45 cases of esophageal cancer. Effects of MSI on clinical stage, pathological classification, lymph node metastasis, and invasion were not found. Conclusion MSI may be an early molecular pathway and play a certain role in the development of the esophageal cancer.

Objective To assess the incidence of thrombocytopenia and abnormal coagulation,door to needle time (DNT),and safety in patients with ischemic stroke who receive intravenous thrombolytic treatment prior to the availability of blood platelet (PLT) and coagulation results.Methods Consecutive acute ischemic stroke (AIS) or transient ischemic attack patients within 12 hours of symptom onset who were admitted from January 2009 to March 2013 were retrospectively recruited.First laboratory reports in their medical charts were collected to assess the incidence of thrombocytopenia and abnormal coagulation.In the mean time,consecutive AIS patients who received intravenous thrombolysis at Huashan Hospital during the same period were retrospectively recruited.The thrombolytic procedures were further optimized since June 10.3760/cma.j.issn.1006-7876.2014.07.00626,2012.With informed consent,intravenous thrombolysis was initiated without PLT and coagulation results after certain previous history and medications were ruled out.Thrombolytic patients were divided into two groups based on the initiation of thrombolysis before or after PLT and coagulation results.Baseline demographic data,symptomatic intracerebral hemorrhage rates,mortality on the 7th day as well as functional outcome at 3 months were collected.DNT as well as efficacy and safety of thrombolysis therapy were compared between the two groups.Results Of 298 AIS patients within 12 hours of onset,8 had thrombocytopenia or abnormal coagulation.One hundred and twenty cases of intravenous thrombolysis patients were recruited.Waiting for PLT and coagulation results prolonged DNT than without waiting for them (90 min vs 59 min; U =870.000,P ＜0.01).There was no statistically significant difference in the rate of symptomatic intracerebral hemorrhage (sICH),early efficacy,7 d mortality and 3-month good outcome between two groups.Conclusions The incidence of thrombocytopenia and abnormal coagulation is low in AIS patients.Initiating intravenous thrombolysis prior to the availability of coagulation results can shorten DNT,while increased risks of sICH and 7 d mortality were not observed.