AIM: To observe the clinical effect of amniotic membrane transplantation and penetrating keratoplasty in the treatment of atheromatous corneal ulcer.
METHODS: Thirteen patients ( 13 eyes ) diagnosed atheromatous corneal ulcer from February 2009 to May 2014 in our hospital were retrospectively analyzed. Surgical treatment including amniotic membrane transplantation and penetrating keratoplasty were used to deal the patients with no effects of drugs. All patients were followed up for 4mo to 2a ( mean 8mo ) after surgery. Visual acuity, healing and recipient of ulcer were examined.
RESULTS:There patients were treated conservatively with corneal ulcer slowly healing, healing time was 14~35 ( 21±12. 1 ) d. Seven cases were treated with amniotic membrane transplantation, 2 cases were treated with penetrating keratoplasty, 1 case of corneal ulcer perforation and lost light perception received enucleation of eyeball. Corneal ulcer were cured in patients performed amniotic membrane transplantation and penetrating keratoplasty. All patients had no recurrence during the follow-up period.
CONCLUSION:Atheromatous corneal ulcer is difficult to be cured by conservative treatment. Most patients need operation treatment. Amniotic membrane transplantation and penetrating keratoplasty can be performed to deal with atheromatous corneal ulcer and abtain satisfactory effect. But amniotic membrane transplantation is relatively simple and easy, and it is suitable for promotion in primary hospitals.

AIM: To investigate the preliminary clinical observation of excimer laser phototherapeutic keratectomy ( PTK ) assisted by anterior segment optical coherence tomography ( OCT ) in the treatment of non II type granular corneal dystrophy.
METHODS: A retrospective case series were studied. Totally 8 patients ( 12 eyes ) who were diagnosed as granular corneal dystrophy underwent PTK from April 2011 to January 2013 in our hospital. All patients were excluded from the II type granular corneal dystrophy ( Avellino corneal dystrophy ) by the Avellino corneal dystrophy rapid diagnostic kit and underwent preoperative anterior segment OCT examination, so as to determine the lesion morphology and depth, and used to guide the setting of PTK parameters. They were followed up for the complications after operation, postoperative recurrence, the recovery of visual acuity.
RESULTS: All patients were followed up for 6-12mo, average 9mo after operation. All patients' best corrected visual acuity were significantly improved, superficial corneal opacity lesions were effectively removed, and the corneal opacity recurrence or serious Haze were not found during the follow-up period after operation.
CONCLUSION: In patients with non type II granular corneal dystrophy, PTK assisted by anterior segment OCT can be accurate, effective removal of corneal lesions, obtain good effect after operation.

The procedures for the calculation and measurement and calibration of output dose of the Siemens primus E accelerator with IAEA regulation are introduced to probe an accurate,rapid and effective method.The parameter value of measurement is determined according to IAEA regulations and absorbed dose calculation formal is briefed to the form that the reading of instrument multiplies exposure calibration coefficient Nx and Air temperature,pressure and humidity effects Ktp and cumulative correction factor Cf the precision of accelerator conform to the standard of QA&QC by the measurement and calibration of the Hospital Primus E accelerator in output dose.The methods of measurement and calibration can be used for the determination of output dose in precise radiotherapy of digital medical linear accelerator and clinical treatment.

AIM:To investigate the changes of connexin 43 (Cx43) via establishing a model of sympathomi-metic atrial fibrillation ( AF) .METHODS:The mongrels ( n=15) were randomly divided into control group , rapid atrial pacing (RAP) group and isoprenaline (ISO) perfusion+RAP group (ISO+RAP group).All mongrels’ hearts were taken out rapidly by median sternotomy to establish the cardiac model with Langendorff perfusion in vitro.The atrial effective re-fractory period ( AERP) and AF inducability were tested .The expression and distribution of tyrosine hydroxylase ( TH) were analyzed by immunohistochemistry .Total protein level of Cx 43 and phosphorylation of Cx 43 were determined by West-ern blot.The distribution of Cx43 were also observed by immunofluorescence staining .The cell apoptosis was analyzed by TUNEL staining.The generation of reactive oxygen species ( ROS) in the mitochondria was measured by fluorescence spec-trophotometry .RESULTS:No significant change of AERP was found between control group and RAP group , while that in ISO+RAP group was significantly decreased (P<0.05) and induced AF.Compared with control group, the expression of TH, apoptotic index and the generation of ROS increased gradually (P<0.05), while the content of Cx43 decreased grad-ually both in the total protein and the phosphorylation levels in RAP group and ISO +RAP group (P<0.05).The fluores-cence intensity of Cx43 was also attenuated and Cx43 were lateralized apparently in RAP group , while Cx43 were character-ized as punctate distribution in ISO +RAP group.CONCLUSION:Sympathetic nerves may activate autophagosome at in-tercalated discs and trigger cell apoptosis , resulting in remodeling and downregulation of Cx 43 via oxidative stress , thus having effects on mediating and maintaining AF .

Objective To determine effects of activating protein kinase C (PKC) on ventricular action potential duration restitution (APDR) and Burst stimulus induced arrhythmia in Langendorff-perfused rabbit hearts.Methods Male rabbits were equally divided into three groups randomly: control group (Tyrode's solution perfusion),PKC agonist phorbol-12-myristate-13- acetate (PMA,100 nmol/L) group and PKC inhibitor bisindolylmaleimide (BIM,500 nmol/L) group.Thirty minutes after perfusion,the monophasic action potential (MAP) and effective refractory period (ERP) were determined in right basal ventricle (RB),right apex ( RA ),left basal ventricle (LB) and left apex (LA) of all the animals,and APDR curve was drawn.Burst stimulus method was used to induce ventricular arrhythmia in perfused rabbit hearts; Real-time PCR was used to detect the mRNA expression of PKC in four different areas of ventricle.Results Compared with the control group,the ERP,90％ of monophasic action potential duration ( MAPD90 ) and ERP/MAPD90 were significantly shortened ( all P ＜ 0.01 ),the max slopes ( Smax ) of APDR curve were significantly steeper (RB:1.22 ±0.23 vs.0.65 ± 0.19 ; RA:2.99 ± 0.29 vs.1.02 ± 0.18 ;LB:1.84 ±0.21 vs.0.85 ±0.12; LA:4.02 ±0.32 vs.1.12 ±0.23,all P ＜0.01 ) and the incidences of ventricular arrhythmia were significantly increased in the PMA group.All parameters were similar between the BIM group and the control group (all P ＞ 0.05).Conclusion Activating PKC could enhance the max slopes of APDR curve at various ventricular areas and subsequently increase arrhythmia susceptibility in Langendorff-perfused rabbit hearts.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have a remarkable differentiation potential and superiority as a type of seed cells,but their application is limited in the presence of certain diseases,such as aplastic anemia and myelogenous neoplasm.The present studies have found that seed cells called muscle-derived stem cells (MDSCs) have brought more and more attention,because of their capability of stir-renewal and multi-diffcrentiation like B MSCs.OBJECTIVE: To explore the biological characterization of the muscle-derived stem cells (MDSCs) from rabbits,and analyze the phenotype.DESIGN,TIME AND SETTING: Cell in vitro observation experiment was performed at the Medical Research Center of Second Affiliated Hospital of Sun Yat-sen University from August 2005 to March 2006.MATERIALS: A New Zealand rabbit (1.5 months old,clean grade) was enrolled for the preparation of Muscle-derived stem cells.Growth medium was DMED-LG added with 10% fetal bovine serum and 10% horse serum and fusion medium was DMEM-LG added with 2% fetal bovine serum.METHODS: The muscle mass was removed from the anesthetized rabbit to isolate MDSCs.These cells were dissociated using three enzymes (collagenase XI,dispase and trypsin) respectively.Sediment was resuspended.Then preplate technique was used.The muscle cell extract was plated on a collagen-coated culture flask with growth medium.The flask was called PP1.PPI was kept overnight in a 37 ℃ incubator containing 5% CO2,After that,the suspension was transferred to another collagen-coated culture flask,which was called PP2.PP3,PP4,PP5 and PP6 were constructed later following the same procedures.The cells adhered in PP6 were collected,plated in 6-well plates,and divided into 2 groups.Growth medium was used in one group,in which the cells were kept growing at a degree of confluence beyond 50%,and fusion medium was used in the other one,in which the cells were passaged with a degree up to 30%.MAIN OUTCOME MEASURES: The cells from PP1 to PP6 were collected,and the characterization was identified preliminary by Flow cytomctry,Immunocytochemistry and Western Blotting.The fusion of cells in PP6 was detected at different confluence degrees and concentration of medium.RESULTS: The cells in PP6 showed ＞ 80% desmin+,＞ 70% Bcl-2+,＞ 95% CD45,which indicated that MDSCs were in a high concentration.The expressions of α-SMA in the cells were decreasing with the Preplate technique used and the cells in PP6 almost had no α -SMA expression.When passaged at a high confluence (＞ 50%) or cultured with low concentrations of serum (2% serum),the cells in PP6 had a strong tendency of fusing into myotubes or cell chains and were skeletal myosin+.CONCLUSION: MDSCs,which are capable of multi-differentiation under a high fusion or low serum conditions,express dcsmin and Bcl-2 highly,but extreruelv little CD45 and no α -SMA.

Objective To establish a method of reverse hybridization to detect five subfamilies of low risk Human Papillomaviruses(HPV6,11,42,43 and 44)and eighteen subfamilies of high risk HPV (HPV16,18,31,33,35,39,45,51,52,53,56,58,59,66,68,73,83 and MM4)in one reaction.Methods Special probes for twenty-three HPV subfamilies were fixed on nylon membrane bars,biotin labeled general primers mediated polymerase chain reaction(GP-PCR)were applied in HPV DNA amplification.PCR amplified DNA fragments were reversely hybridized with special probes that were fixed on the membranes. All samples(136)detected by reverse hybridization method were paralleled with the methods of Hybridization Capture Ⅱ(HC-Ⅱ)and sequencing.Results Positive rate of the 136 samples detected by reverse hybridization was 41.9%,while HC-Ⅱ 42.6% and sequencing 40.4%.Reverse hybridization detection indicated coherence with the other two methods(Kappa 0.8644 and 0.9089,respectively).While sequencing was lab standard for DNA test,the sensitivity was 96.36%,specificity was 95.06%,accuracy was 95.59%.Conclusions Method of reverse hybridization is adaptable to 23 kinds of HPV subfamilies, which can confirm the exactly subfamilies of HPV infection.This method is adaptable in clinical detection of HPV,with high sensitivity,high specificity,simply and convenient operation and the results are easily to be read.

Objective: To investigate the effects of vitamin B 1, C and orange juice on human lung cancer cell proliferation in vitro. Methods: Vitamin C degradation in culture medium was evaluated. The methods included: methy thiazolyl tetrazolium (MTT) assay, colony forming assay and 3H TdR incorporation test. The final concentrations of factors in medium were: orange juice (vitamin C 30,60,120 ?g/ml), vitamin C(30,60,120 ?g/ml), vitamin B 1 (0.01,0.1,1 ?g/ml). Results: (1) In MTT test, orange juice at each level had significant inhibitory effect on the growth of lung cancer cells (P

AIM To study the effect of ketamine on proliferation,cell cycle in the cultured rat neural stem cells. METHODS The growth inhibition of ketamine on neural stem cell was evaluated by an MTT assay. The effect of ketamine on cell cycle was measured by flow cytometry. RESULTS Ketamine inhibited the growth of cultured rat neural stem cells. Flow cytometry analysis showed that G 0/G 1 phase rate was increased but S phase rate was decreased. CONCLUSION Ketamine can inhibit proliferation of cultured rat neural stem cells,and this inhibitition is associated with cell cycle block.

In this study, in vitro chemosensitivity testing was conducted on primary cultured breast cancer cells from 96 patients with breast cancer, and the results showed that the cells from a few patients with primary breast cancer developed multidrug resistance (MDR) prior to the first chemotherapy exposure. All the cells from the recurrent cancer patients had MDR. The findings suggested that patients having MDR would benefit from high-dose chemotherapy (HDC) regimens. In vitro chemosensitivity screening, which was aimed at improving the therapeutic efficacy and minimizing side effects, helps in choosing individualized treatment for breast cancer.
Antineoplastic Agents/*pharmacology ; Breast Neoplasms/*drug therapy ; Breast Neoplasms/pathology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Drug Screening Assays, Antitumor/*methods ; Neoplasm Recurrence, Local/*drug therapy ; Tumor Cells, Cultured