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.

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.

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.

This paper is to explore changes of intestinal mucosal barrier, intestinal flora, and bacterial translocation in rats with severe acute pancreatitis (SAP). Twenty four male SD rats were randomly divided into the control group (n = 10) and the experimental group (n = 14). The model of severe acute pancreatitis of rats was induced by the method of injecting adversely 5% sodium taurocholate into the common biliary-pancreatic duct. All of the rats were killed after 24 hours and the level of the serum amylase and the plasma endotoxin was determined after that. The pathological changes of pancreas and small intestine were observed through hematoxylin-eosin staining (HE staining) and the abdominal viscera bacterial translocation rates were tested. With the method of real-time polymerase chain reaction (RT-PCR) the quantity of the intestinal flora was analyzed. In the control group, the level of Escherichia coli, Lactobacillus and Bifidobacterium were 2.08 ± 1.29, 11.04 ± 7.55 and 12.21 ± 4.95, respectively. On the contrast, the level of Escherichia coli in the cecum contents was much higher (9.72 ± 3.58, P < 0.01), while the Lactobacillus number was decreased significantly (0.67 ± 0.34, P < 0.01), and the Bifidobacterium number was also decreased (4.59 ± 3.42, P < 0.05) in the experimental group, so the ratio of Bifidobacterium/Escherichia coli was reversed. Besides, in the experimental group, the plasma endotoxin positive rates and the bacterial translocation rates were much higher (P < 0.01 or P < 0.05) and the pathology scores of pancreas and small intestines were also significantly higher (P < 0.01) than those in the control group. These results indicated that in severe acute pancreatitis rats, the intestinal mucosal barrier was severely damaged and the dysbacteriosis occurs in the intestinal canal. And these might relate to the occurrence and development of multiple organ infection.
Animals ; Bacterial Translocation ; Endotoxins ; Intestinal Mucosa ; pathology ; Intestines ; microbiology ; Male ; Pancreas ; pathology ; Pancreatitis ; microbiology ; pathology ; Rats ; Rats, Sprague-Dawley

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 observe the effect of forearm crutches on motor function of children with spastic cerebral palsy. Methods 60 children with spastic cerebral palsy were divided into observation group (n=30) and control group (n=30). The control group accepted rou-tine rehabilitation, while the observation group were also trained to use forearm crutches. They were assessed with Gross Motor Function Measure-88 (GMFM-88) and Balancer. The way of item 70 of GMFM-88 was used to assess the mobile capability. Results The scores of GMFM-88 significantly improved in both groups after treatment (t>6.002, P<0.001), and improved more in the observation group than in the control group (t=2.317, P<0.05). The whole path length and the circumference area reduced in both groups after treatment (P<0.05), and reduced more in the observation group with the assist of the forearm crutches (P<0.01). The incidence of walking was more in the observa-tion group with the assistant of the forearm crutches (χ2=25.87, P<0.01). Conclusion Forearm crutches assistant can improve the recovery of motor function, balance and walking ability in children with spastic cerebral palsy.

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.