Objective: To establish the quality standard for Tiaolitongbao I capsules.Methods: The components including Astragalus , Atractylodes, Finger citron and Radix aucklandiae were identified by TLC.Emodin, the effective component of Polygonum cuspidatum , was determined by HPLC.Results: The characteristic spots in TLC were clear without any interference.The linear range of emodin was 4.25-68.00 μg·ml-1 (r =0.999 9).The average recovery was 95.22% ,and RSD was 1.47% (n =6).Conclusion: The methods used for the identification and quantification are sensitive, simple and accurate, which can be used for the quality control of Tiaolitongbao I capsules.

AIM: To study the quality standard of Compound Shencha Granuls (Radix et Rhizoma Glycyrrhizae, Radix Rehmanniae, etc.). METHODS: Radix et Rhizoma Glycyrrhizae and Rhizoma et Radix Polygoni Cuspidati in Compound Shencha Granule were distinguished by TLC. Ursolic acid was determinated by TLCS. RESULTS: The methods of TLC to distinguish Radix et Rhizoma Glycyrrhizae and Rhizoma Polygoni Cuspidati were feasible without negative inference. Ursolic acid content in three groups of samples was 1.24 - 1.58 mg?g -1 . CONCLUSION: The method is accurate and reproducible. It can be used for the quality control of Compound Shencha Granule.

OBJECTIVETo investigate the main factors that influence the results of sperm alkaline single-cell gel electrophoresis (SCGE), optimize the conditions, and standardize its procedures.

METHODSUsing alkaline SCGE, we detected the DNA fragments of sperm treated with different concentrations of H2O2 and determined the influences of the number of agarose gel layers, pH during DNA unwinding and electrophoresis, the time of DNA unwinding and electrophoresis, and cumulative sperm number on the results of sperm alkaline SCGE. Then we optimized the procedures, analyzed the repeatability of the optimized method, and examined 40 semen samples using the method.

RESULTSThree agarose gel layers could reduce the background. The optimal pH during DNA unwinding and electrophoresis was 10, and the best times for DNA unwinding and electrophoresis were 40 min and 30 min, respectively. Fifty sperm were adequate to ensure the reliability of the results. Based on the percentage of tail DNA, the intra- and inter-assay repeatabilities of the optimized sperm alkaline SCGE were 3.12% and 7.13%, and by the DNA damage score, they were 2.38% and 6.09%, respectively. Sperm DNA fragments were significantly increased in the infertile patients with oligoasthenoteratozoospermia as compared with healthy fertile males (P <0.05).

CONCLUSIONThe optimized sperm alkaline SCGE, highly repeatable and easy to be standardized, can be applied to the clinical detection of sperm DNA fragmentation in infertile men.

Asthenozoospermia ; genetics ; Comet Assay ; standards ; DNA Damage ; DNA Fragmentation ; Humans ; Hydrogen Peroxide ; toxicity ; Male ; Oligospermia ; genetics ; Oxidants ; toxicity ; Reproducibility of Results ; Sperm Count ; Spermatozoa ; drug effects ; enzymology ; Time Factors

Objective To explore the differences of emotion identification and emotion quotient (EQ) scores between the teenage criminal group and the vocational school student group,and conduct research on the correlation between the emotion identification and EQ of teenage criminals.Methods A self-designed questionnaire,experiments about emotion identification and EQ tests were applied to 48 teenage criminals and 46 vocational school students who had the matching age,IQ and sex with the teenage criminals.Results The number of correct responses to neutral mood among the teenage criminals (60.00± 17.07) was lower than that of vocational school students (66.12±5.45).There was no difference about EQ between these two groups(90.48± 13.31,90.76± 19.85,P ＞0.05),but in pressure management dimensions,teenage criminals had some differences compared with the vocational school students (53.97± 8.95,57.84 ± 7.26,P＜ 0.05),especially in impulse control (24.97 ± 4.98,28.95 ± 5.22,P＜0.01).The general state of mind,pressure management,individual components and interpersonal components was related to the emotion identification(r 1 =0.43,r 2 =0.36-0.38,r 3 =0.37,P＜0.05).Conclusion Emotion identification of teenage criminals have some flaws,especially the judgment on neutral mood,and it is easily to be wrong.Teenage criminals lack of controlling in EQ.The emotion identification and EQ of the teenage criminal interact with each other.

Objective To investigate the clinicopathologic features and prognostic significance of BRCA1 and Ki-67 expression in breast cancer.Methods The expression levels of BRCA1 and Ki-67 were assayed by immunohistochemistry in 194 cases of breast cancer tissues.The correlations of BRCA1 and Ki-67 expression with patients' clinicopathologic features were also analysed.Results Low expression of BRCA1 was detected in the lymph node metastasis group,ER/PR negative group,and HER-2 positive group in 194 patients with breast cancer (P ＜ 0.05),as well as in the triple negative breast cancer (TNBC) group compared to non-TNBC group (P ＜0.05).High expression of Ki-67 was detected in patients with higher histological grade,negative ER/PR,and positive HER-2 (P ＜ 0.05).Furthermore,negative correlation was found between the expression of BRCA1 and Ki-67 (P ＜ 0.05).The combination of low expression of BRCAl and high expression of Ki-67 was mostly found in the patients with postmenopausal,lower histological grade,lymph node metastasis,negative ER/PR and positive HER-2 (P ＜ 0.05).However,there was no significant difference between TNBC and non-TNBC.Conclusion Joint detection of BRCA1 and Ki-67 might play an important role in predicting clinical outcomes of breast cancer,especially BRCA1 may be one of prognostic factors in TNBC.

Objective To study interleukin-1 receptor(IL-1R) and connexin(Cx36) gene expression following Mg 2+-free-induced seizures in cultured developing neuron. Methods Rat embryo cortical neurons cultured for 6 and 17 days were exposed to Mg 2+-free media to induce seizure. At different time after Mg 2+-free treatment, real-time RT-PCR was used to detect IL-1R and Cx36 mRNA expression. Results 1. IL-1R mRNA expression transiently decreased after Mg 2+-free treatment in neurons cultured for 6 and 17 days in vitro. Then the levels of IL-1R mRNA expression recovered in neurons cultured for 6 days, but IL-1R mRNA expression were increased in neurons cultured for 17 days compared with control group and the peak was at 24 hours. 2. In neurons cultured for 6 days in vitro, Cx36 mRNA expression increased after Mg 2+-free treatment compared with control group, the peak was at 24 hours. But in neurons cultured for 17 days in vitro, Cx36 mRNA expression decreased at 6 hours after Mg 2+-free treatment compared with control group, the peak was at 24 hours. Conclusions IL-1R mRNA and Cx36 mRNA expression following Mg 2+-free-induced seizures are different between the neurons cultured for 6 and 17 days in vitro. This is possibly related to the different neuron injury between 6 and 17 days in vitro following seizures.

Objective To observe the energy consumption of children with cerebral palsy before and after assisted walking. Methods From January, 2014 to October, 2015, 21 children in primary school of Grades 1~5 (control group) and 22 children with cerebral palsy in our hospital (observation group) were enrolled. They were required to walk in the 50 meters trail for six minutes. The resting heart rate, the walking distance and the heart rate after walking were measured, and the walking speed and the physical consumption index (PCI) were cal-culated. The observation group was tested with and without forearm crutches. Results Compared with the control group, the walking dis-tance and speed significantly decreased (t>10.653, P<0.001), and the PCI significantly increased (t>4.207, P<0.001) in the observation group. For the observation group without forearm crutches, the heart rate after walking, and the difference of the heart rate significantly de-creased (t=8.389, P<0.001), and the walking distance, walking speed and PCI decreased (t>2.382, P<0.05) when they walked with forearm crutches. Conclusion Assisted walking can decrease the energy consumption of children with cerebral palsy.

Objective To explore the application of gas chromatography mass spectrometry (GC/MS) in children with high risk of inher-ited metabolic disease. Methods From March, 2010 to November, 2015, 119 children suspected with inherited metabolic disease were in-cluded. The urinary organic acid was detected with GC/MS, and related diseases were screened. Results Seventeen children (14.29%) were positive with inherited metabolic disease, in which 16 cases (94.12%) manifested with development retardation. 20 children (16.81%) were probable positive. Conclusion GC/MS is effective in screening children with high risk of inherited metabolic disease, which can provide ba-sis for further diagnosis.

Objective To discuss the distance training for medical staff of children's rehabilitation. Methods 15 lessions were carried out using computer software from April, 2013 to November, 2014 in our centre. 7 cooperative hospitals participated the trainings. The implemen-tation effect was surveyed after training. Results 2693 person-times attended the trainings, including 2109 person-times with living distance training and 584 person-times with video distance training. In the following sampling survey, 92.8%staff thought that the distance training was helpful to their work. 98.6%staff thought that the living distance training was better than the video distance training and 88.6%staff thought their rehabilitation level improved after the distance training. Conclusion The distance training of children's rehabilitation has the advantages of economy, convenience and celerity. It has a wide developmental prospect.

Objective To knock out Asxl2 gene in murine embryonic fibroblast cell line NIH3T3 using CRISPR/Cas9n system. Methods A pair of sgRNAs which targeted exon 5 of Asxl2 gene were designed and subcloned into the pX462 vec?tor. The recombined plasmids were verified by sequencing and transfected into NIH3T3 cell line. Single cells were isolated through serial dilutions, followed by an expansion period to obtain new monoclonal cell lines. The genomic DNA of the new monoclonal cell lines was extracted and a DNA fragment flanked the target site was amplified by genotyping PCR then se?quenced. Lastly, western blotting were applied to confirm whether Asxl2 was successfully knocked out. Results The CRIS?PR/Cas9n plasmids that targeted Asxl2 were successfully constructed. NIH3T3 cells were co-transfected with the two recom?binant constructs. After puromycin selection, subclonal cell lines were obtained and one of them was validated by genotyping PCR-sequencing. Western blotting also confirmed that Asxl2 was completely depleted in the NIH3T3 cell line. Conclu?sion CRISPR/Cas9n plasmids that targeted Asxl2 were successfully constructed therefore a Asxl2 knockout NIH3T3 stable cell line was established via this system.