Objective To explore the status of C57BL/6J mouse brown fat adipogenic differentiation function with aging.Methods C57BL/6J female and male mice at the ages of 0-week (newborn),4-week,8-week,12-week old were selected from the same brood,brown adipose tissue was obstained from their interscapular region,and the brown adipose was identified by using immunohistochemical markers.Then the total RNA was extracted from the brown adipose and quality identification was determined at the same time.The expression levels of the related genes (PPARα,C/EBPα,PGC-1α,PPARγ,FOXC2,BMP7) induced by brown adipose adipogenic differentiation were detected by quantitative real-time PCR in 0-week,4-week,8-week,12-week mice.Results Uncoupling protein -1 (UCP1) immunohistochemical data indicated that positive deep-colour substance was brown adipose tissue.Quantitative Real-time PCR also indicated that the expression volume of adipogenesis gene gradually reduced with aging,and there were significant differences at the different time points [PPARα (F =11.96,P ＜ 0.000 1),C/EBPα (F =9.39,P ＜0.000 1),PGC-1α(F =17.21,P ＜0.000 1),PPARγ(F =13.11,P ＜0.000 1),FOXC2(F =12.23,P ＜0.000 1),BMP7(F =16.44,P ＜0.000 1)].Conclusions The adipogenic differentiation ability and activity of mouse brown adipose gradually reduce with aging.But the regulatory factors for its function needs to be further investigated.

Objective To investigate the relationship between standard CD44(CD44s)expression and tumor stage and prognosis in colorectal cancer.Methods CD44s expression was detected by immunohistochemistry in tumor tissues and normal mucosa specimens from 74 cases of primary colorectal cancer.Results Of 74 cases,the expression of CD44s in colorectal cancer tissue and normal mucosa specimens was documented in 42% and 16%,respectively.The expression of CD44s in primary tumors significantly increased in stage Ⅲ and Ⅳ than in stage Ⅰ and Ⅱ(39% vs.6%,x~2=8.46,P＜0.01).A statistically significant correlation was observed between the overall survival and CD44s expression(x~2=17.82,P＜0.01).Furthermore,a poor prognosis of CD44s-positive tumors was observed in patients with stage Ⅲ and Ⅳ colorectal cancer(x~2=16.23,P＜0.01),but not in patients with stage Ⅰ and Ⅱ colorectal cancer(x~2=1.34,P＞0.05).Multivariate analysis indicated that TNM stage and CD44s expression were independent predictors of overall survival in colorectal cancer.Conclusion CD44s overexpression is involved in the progress of colorectal cancer and predicting the prognosis.

In order to make the experimental teaching adapt to the development of modern teaching idea and to meet the needs of the pharmaceutical industry for high quality talents, pharma-cology experiment teaching method has been reformed. Single teaching method has turned to diversi-fied teaching method according to the experimental contents, difficulty and characteristics of teaching situation, such as leading method of using theory after experiment in validation experiments and single blind method in multidrug efficacy experiment in the early stage, case-based teaching in comprehensive experiments in the middle stage , and student teaching method in designing experiments in the later stage. The study results show that students' interest in learning has been inspired,their experimental enthusiasm has been mobilized,and their operation, analysis and problem-solving ability has been improved, which is advantageous to the comprehensive quality education.

Objective To identify biomarkers associated with the differentiated phenotype based on gene expression profiling of gastric cancer. Methods Two bioinformatic methods, BAGEL and k-TSP, were used to identify featured genes associated with differentiation in gastric cancer samples based on the Oligo gene chip data, and ROC curves were used to verify the classification sensitivity and specificity of the identified genes. Finally, a total of 30 gastric cancer samples with different differentiation levels were collected for laboratory validation using real-time PCR analyses. Results A total of 121 differentially expressed genes were identified using the BAGEL algorithm, the criterion were FC ＞ 2. 0 and P ＜ 0. 001.Then, the k-TSP algorithm for feature selection based on this differential expression data were used, and 3 groups of featured genes which had potential to classify poor and well differentiation gastric cancer samples were identified, including MYLIP and TMPRSS3, ZNF266 and TM4SF1, SNAI2 and CNFN. To define the featured gene groups that had the highest classification capability, ROC curves to calculate the classification sensitivity and specificity of each gene group were used. The results showed that the combination of SNAI2and CNFN as a classifier had the highest classification sensitivity and specificity. Real-time PCR results showed that 18 of 22 poor differentiation samples were found with high expression of SNAI2 and low expression of CNFN (82%); 6 of 8 well differentiation samples were of low expression of SNAI2 and high expression of CNFN (75%). Conclusion The results indicate that SNAI2 and CNFN are constantly expressed in poor or well differentiation gastric cancer samples, and the expression pattern of these two genes is opposite. These results indicate that SNAI2 and CNFN have the potential for the identification of the differentiation level of gastric cancer.

Objective To investigate the neuroprotective effect of Benztropine on retinal ganglion cells (RGCs) death and optic nerve injury in rats model of non-arteritis anterior ischemic optic neuropathy (rNAION).Methods A total of 25 Sprague-Dawley rats were randomly divided into Benztropine treatment group (n=13)and PBS control group (n=12).The right eye was set as the experimental eye.rNAION model was established by using rose Bengal combined with laser photodynamic method.The rats in the Benztropine treatment group were received intraperitoneal injection with Benztropine 10 mg/kg (0.2 ml) daily for 3 weeks,while the rats in the PBS control group were received intraperitoneal injection with an equal volume of PBS.At 1,3 and 7 days after modeling,the retinal and optic disc conditions of the rats were observed by direct ophthalmoscopy.Retrograde labeling,fluorescence microscopy and transmission electron microscopy were used to observe the survival of RGCs and the damage of the optic nerve myelin and axon at 4 weeks after modeling.The RGCs density and survival rate of the two groups were compared by One-Way Anova.Results At 1 and 3 days after modeling,the optic disc edema was observed in the rats of rNAION model group.At 7 days after modeling,the optic disc edema decreased and the boundary was blurred compared with 3 days after modeling.After 4 weeks,the RGCs density in the PBS group was 308± 194/mm2 and the survival rate was 13.7％.The density of RGCs in the Benztropine group was 1173+868/mm2 and the survival rate was 47.6％.The differences of RGCs density and survival rate were significant between the two groups (F=7.552,8.184;P=0.015,0.012).Myelin disintegration,axon degeneration,onion-like body and gliosis were observed in the optic nerve sections of rNIAON in the PBS group,while the damage ofaxon and myelin structure in the Benztropine group was significantly less than that in the PBS group.Conclusions Benztropine group showed higher RGC survival rate,less damage ofaxon and myelin structure on rNAION model.This study explored the potential neuroprotective effect of Benztropine.

Objective:To evaluate the safety and efficacy of 27G micro-incision vitrectomy surgery (MIVS) combined with intravitreal injection of ranibizumab (IVR) in the treatment of retinopathy of prematurity (ROP) with early intervention failure.Methods:Retrospective case series was performed. Fourteen eyes (11 infants) with ROP who underwent 27G MIVS combined with IVR were included from March 2016 to January 2018 in Shenzhen Eye Hospital. Among them, there were 5 males with 7 eyes, 6 females with 7 eyes. The average gestational age of the infants was 28.12±0.90 weeks; the average birth weight was 1 023.64±200.96 g. Before the early clinical intervention, 1 infant (2 eyes) had ROP in zone Ⅰ stage 3 with plus disease, 8 infants (10 eyes) had ROP in zone Ⅱ stage 3 with plus disease, and 2 infants had ROP in aggressive posterior ROP. Six eyes underwent laser photocoagulation, while 8 eyes received laser therapy combined with IVR. Six eyes of stage 4A ROP and 8 eyes in stage 4B. Retinal detachment was detected with a mean of 10.44±9.21 weeks. At the time of surgery, the average post-conceptional age was 48.02±8.09 weeks. All the affected eyes were treated with standard sclera with three incisions 27G MIVS. During the operation, only local vitrectomy was performed to release and clear fibroascular proliferation in the optic disc, anterior macular area and pericristal area. After surgery, 10 mg/ml of ranibizumab 0.03 ml was injected into the vitreous cavity. The average follow-up time was 23.36±8.34 months. The primary objectives were the condition of retinal reset, ROP progression control and complications.Results:All patients had uneventful surgeries with an average duration of 32.86±9.35 mins. Of the 14 eyes, 12 eyes (85.71%) were controlled, 8 eyes (57.14%) had a good rearrangement of macular structure, while 4 eyes with macular traction. Two eyes had ROP progression, recurrence of retinal detachment, posterior synechia. Complicated cataract was in 1 eye. Proliferative vitreoretinopathy and retinal detachment was in 1 eye after 7 months the operation.Conclusion:27G MIVS combined with IVR is a safe and effective treatment for ROP with early clinical intervention failure.