BACKGROUND:Kunming mouse embryonic fibroblasts are the most common feeder layers at present, and there are rare reports addressing C57BL/6 mouse embryonic fibroblasts as feeder layers.
OBJECTIVE:To separate and culture C57BL/6 mouse embryonic fibroblasts in vitro, and produce feeder layers to enlarge the resources of mouse embryonic fibroblasts.
METHODS:C57BL/6 mouse embryonic fibroblasts were isolated and cultured by trypsin digestion method in vitro. The biological characteristics and growth rule of the fibroblasts were investigated, then the feeder layers for the cel culture were produced. The growth of cel colonies on the prepared feeder layer was tested.
RESULTS AND CONCLUSION:C57BL/6 mouse embryonic fibroblasts grew wel with a large amount, by trypsin digestion method at different concentrations. There was no significance in the survival rate after cryopreservation for 1 week, 2 weeks, 1 month, 3 months and 6 months. The cel s were proliferative from the second to fifth passage and declined sharply after the sixth passage. The planted mouse embryonic fibroblasts feeder layers had a high activity within 3 days, but got a sharp decline after 4 days. So it is best to use C57BL/6 mouse embryonic fibroblast feeder layers within 3 days after they’re inactivated. C57BL/6 mouse embryonic fibroblast feeder layer can support embryonic stem cel s and induce pluripotent stem cel s to grow as Kunming mouse embryonic fibroblasts.

Objective:To observe the feasibility of promoting blood circulation by removing blood stasis in target interfering ADUB endometrial hyperplasia.Methods:Endometrial hyperplasia model was established,and the rats were divided into normal group(group A),model group(group B),small,middle and large dosage groups of Chinese drugs(C group,D group,E group,respectively).To observe uterus humid weight,uterus index,morphological change of endometria and expression of VEGF before and after treatment.In addition,30 cases of Anovulatory dysfunctional uterine bleeding were selected and given intrauterine injection of Qumozhibeng Cream twice in the first week after menstrual period,the change of menstrual blood amount were observed after applying medicine for one month,three months and six months.Results:After treatment,the uterus humid weight and uterus index decreased(P

Objective To investigate the clinical application of ultrasound biomicroscopy in the treatment of congenital corneal opacities.Methods Medical records of 20 eyes (15 patients) with congenital corneal opacity treated at our hospital from July 2004 to November 2011 were retrospectively reviewed.Best corrected visual acuity testing,intraocular pressure testing,slit-lamp anterior segment examination,fundus examination,slit-lamp microscopic photography,B scan examination,and ultrasound biomicroscopy were performed for analysis of complications of congenital corneal opacity and selection of surgical approaches.Results The ultrasound biomicroscopic examination showed that 5 eyes had no Descemet's membrane and corneal endothelium,20 eyes had anterior synechia,5 eyes had aniridia,3 eyes had loss of lens cortex,13 eyes had cataract,14 eyes had closed angle,and 3 eyes had pupillary membrane.14 of 20 eyes received surgical treatment,including penetrating keratoplasty combined with cataract extraction and trabeculectomy (5 eyes),penetrating keratoplasty combined with pupil angioplasty (3 eyes),penetrating keratoplasty combined with cataract extraction (3 eyes),penetrating keratoplasty combined with trabeculectomy (2 eyes),and lamellar keratoplasty (1 eye).Conclusions Ultrasound biomicroscopy is important to guide the diagnosis and treatment of congenital corneal opacity.

AIM:To study the human brain hemodynamics responses induced by acupuncturing foot shaoyang gallbladder meridian point GB37 and visual stimulation,and to analyze the effect of acupuncture to activate the cerebral functional area in visual stimulus. METHODS:Twenty five normal subjects were divided into two groups:15 subjects for electro -acupuncture GB37 in visual stimulus group and 10 subjects for electro-acupuncture GB37 groups. The blood oxygen level dependent functional magnetic resonance imaging (BOLD-fMRI) was carried out under the condition of two dif-ferent stimulation patterns (including a vision stimulation task and point GB37 acupuncture under the vision stimulation condition). A scintillating chessboard grid was applied as the vision stimulation task and the frequency was 8 Hz. The activation of cerebral functional areas of tested people under the conditions of the vision stimulation task and acupuncture were analyzed,respectively. RESULTS:Out of 15 subjects of electro -acupuncture GB37 in visual stimulus groups,twelve health volunteers were enrolled. When the vision stimulation task was carried out,the alarge area of brainstem and posterior lobe of cerebellum was seen to be activated. The areas of limbic lobe,hippocampus,middle temporal gyrus and inferior temporal gyrus were also activated. Under the condition of acupuncture only,the areas of posterior central gyrus,inferior parietal lobule,parietal lobe precuneus and the middle frontal gyrus were activated. When doublesided points of gallbladder meridian of foot-shao-yang were electroacupunctured under the visual stimulus status,the areas of superior frontal gyrus,inferior frontal gyrus,parietal precuneus and occipital gyrus lingualis were activated. CONCLUSION:The functional MRI shows the activation of cerebral functional area after acupuncture. Visual stimulus causes significant changes of brain conduction path connection. Under the visual stimulus status,the acupuncture to the points of gallbladder meridian of foot -shao -yang,GB37 has an extent of influence to conduction path conformity of the vision system and with the treat-ment of ophthalmocace. Furthermore,the external stimulation has prominent influence to the acupuncture therapy possibly.

AIM: To establish HPLC fingerprint of Venenum Bufonis in an attempt to become a standard of quality control. METHODS: The HPLC method was set up using Alltima C_(18)(250 mm?4.6 mm,5 ?m) column with mobile phase of acetonitrile-0.5% potassium dihydrogen phosphate solution of water;UV detection wavelength at 296 nm and column temperature at 30 ℃ with the flow rate of 1.0 mL/min;20 ?L of the injection volume. RESULTS: In this chromatogram condition,10 peaks were identified as the characteristic fingerprints of Venenum Bufonis.All samples showed the content differences among the samples.The retention times for resibufogenin、cinobufagin、bufalin、bufotalin and cinobufotalin in Venenum Bufonis were consistent with each other.The fingerprint showed good similarity up to 93% in samples from different habitats. CONCLUSION: The method is exact、simple and accurate,and can be used for the identification and quality control of Venenum Bufonis.

OBJECTIVE: To evaluate the clinical efficiency of recombinant adenovirus p53 injection (rAd-p53) combining chemotherapy in the treatment of malignant body cavity effusion. METHODS: 50 cases with malignant body cavity effusion were randomly divided into 2 groups. The treatment group were given intracavitary administration of rAd-p53 1 ? 1012VP after puncture drainage, 48h later which were given intracavitary administration of 60mg/m2 cisplatin once a week for 3～4 weeks. The control group was given the same intracavitary therapy as the treatment group but without rAd-p53 therapy. RESULTS: The total effective rates of the treatment group and the control group were 85.7% and 51.7%(P

To prepare monoclonal antibody of carbohydrate antigen 19-9(CA19-9).Methods: Based on the titer test results of mouse ascites and its IC 50 values ,the mouse that prepare for fusion was identified.Positive monoclonal cell strains were established by cell fusing and screening.Monoclonal antibody from ascites was produced by peritoneal injection monoclonal cell , and then purified by octoic acid-ammonium sulfate precipitation method.After determine the protein concentrations by UV-spectrophotometry ,the monoclonal antibody against CA 19-9 was labelled with horseradish peroxidase.Based on antibody pairing test , DAS-ELISA method was established .To compared with abroad kit , analyzing performance of this method.Results: Three strains of monoclonal antibody were obtained.And the optimal working concentrations of mAb (ZJY3-1G9) ,as coated antibody,McAb(ZJY2-7F10),as HRP-IgG,were assured.Limit of detection was 26.4 U/ml.Linear range was 30-300 U/ml.By detecting patients with serum 33 , confirmed the correlation coefficient of r=0.950 4 , compared with abroad kit that measure simultaneously.Conclusion:Monoclonal antibody prepared for CA 19-9 can be used to develop a kit.

Objective To evaluate the value of whole genome amplification (WGA) combined with array comparative genomic hybridization (aCGH) in prenatal diagnosis. Methods Array CGH were performed by the DNA of 18 prenatal specimens , which were amplified by WGA because of the low DNA yield. Result 3 of the 18 fetuses were 45, X0 and 9 of 15 fetuses with normal aCGH results showed healthy outcome. Conclusion It’ s feasible for prenatal diagnosis using WGA combined with aCGH which not only can shorten the reporting time but also keep the sensitivity and accuracy of detection.

Objective:To compare the durability of resin-based root-surface coating material and all-in-one self-etching adhesive on root surface in vitro.Methods:Human extracted premolars or molars with intact roots were selected.The cementum was removed using a periodontal scaler to expose root dentin. The root surface was coated with an acid-resistant nail varnish,leaving a window of 3 mm ×3 mm on the exposed dentin.The window was covered with either PRG Barrier Coat (PRG)or Clearfil S3 Bond (CS3).After water aging for 14 d,specimens were immersed in acid buffer at pH 4.5 for 4 d and the demineralization buffer was changed every 24 h.Then the specimen was split longitudinally through the center of the ‘window’and the cross-sectional surface was observed with scanning electron microscope (SEM).After fixed and dehydrated,the prepared samples were coated with platinum.The coating mate-rial,root dentin and the interface was observed by scanning electron microscope (SEM).The thickness of the coating material was measured on the SEMimages.Regarding toothbrush wear test,coronal dentin-disks were prepared and covered with PRG and CS3,respectively.After storage in water for 24 h,the specimen was subjected to the toothbrush wear tester for 100,200,300,500,700,1 500 brushing cy-cles.A slurry of fluoride toothpaste (1 ∶2 ratio of toothpaste and deionized water by weight)was used and the brushing load was 300 N.The surface microstructure of remaining coating material was analyzed using SEM.The wear depths were determined by a profilometer.Statistical analysis was performed with SPSS 20.0 by one-way ANOVA.The level of significance was at 0.05.Results:Application of PRG Barrier Coat produced a coating layer of (47.1 ±27.3)μm,while CS3 presented a thin film of (5.7 ± 2.1)μm in thickness.The exposed dentin was hermetically sealed and no obvious gap was observed at the interface in both PRG and CS3 groups.There was no dentin demineralization observed in both groups after water aging.The wear depths of PRG and CS3 increased along with the numbers of brushing cycles. PRG wore at a significant lower pace than CS3 did (P <0.05).Conclusion:PRG coating resin had similar performances as CS3 on protecting root dentin from demineralization after water aging.What’s more,PRG demonstrated a higher toothbrush wear resistance than CS3.We concluded that PRG Barrier Coat contained S-PRG filler may be an effective coating material for protecting exposed root from both chemical and mechanical challenges.Further studies should be carried out to evaluate the long-term reli-ability of the rootsurface coating materials under the clinical setting.

Objective To determine the reproductive physiology and blood physiological and biochemical characteristics of SPF Yorkshire and Landrace swine.Methods Ten reproductive physiology parameters,19 blood physiological parameters and 18 blood biochemical parameters in SPF Yorkshire and Landrace swine were measured using conventional methods and the differences between population,between age groups and between both sexes were analyzed.Results There were no significant differences(P>0.05) in reproductive physiology parameters and most blood physiological and biochemical parameters of the SPF Yorkshire and Landrace swine.A few of parameters,such as blood physiological indices GRAN,HGB,RDW,PLT,PCT,and blood biochemical indices ALKP,CHOL,TBIL,BUN,showed significant difference(P<0.05) between populations,between age groups and between both sexes,however,the values of difference were rather small,deviated from the normal range.Conclusion The physiological and biochemical characteristics of SPF Yorkshire and Landrace swine are basically stable and there is no significant difference compared with other laboratory miniature pigs.This study will provide valuable basic data for raising velvet yield,establishment of animal models and evaluating the genetic quality of closed colony.