Construction of severe combined immunodeficiency mice based on CRSIPR/Cas9 technology / 中国实验动物学报

Objective To knockout Rag2 and IL2rg genes and construct severe combined immunodeficiency mice based on CRISPR/Cas9 technology. Method Design and synthesis of 25 bp sgRNA were made according to the Rag2 and IL2rg sequences in Genbank. After annealing, sgRNA was cloned into pX330 vector. Recombination plasmid Rag2?sgRNA, IL2rg?sgRN and Cas9 were then transcribed into RNA, these RNA were microinjected into zygotes and the zygotes were transplanted into recipient ICR mice. F0 founders were born and mutated F0 founders mated with wild type mice to obtain F1 generation heterozygous mice. Mutated F1 mice were crossed and got F2 generation homozygous mice. Genotype and phenotype of the knockout mice were identified by sequencing, flow cytometry and xenograft model. Results Rag2?sgRNA and IL2rg?sgRNA recombination plasmids were constructed and transcribed into RNA. After microinjection and mat? ing, F0 founders were born and F2 homozygous mice were obtained. The results of sequencing showed that there were two types of genotype in IL2rg gene, 10 bp or 11 bp deletion;however, there was only one genotype in Rag2 gene, which was 8 bp deletion. Compared with wild?type BALB/c mice, the number of CD3 +, B220 + and NKp46 + cells in peripheral blood of the knockout mice was reduced significantly. After inoculation of human breast cancer cell line SKBR?2HL cells, tumor size in the xenograft mouse model was increased gradually along with time extension. Conclusion CRISPR/Cas9 is an efficient way to mutate Rag2 and IL2rg gene in mice in vivo, leading to aberrant T cells, B cells and NK cells.

The application of two enterostomy in gerontal patients with obstructive carcinoma of sigmoid and high rectum / 实用医学杂志

Objective To explore the safety and efficacy of loop ileostomy and Hartmann operation in gerontal patients with obstructive carcinoma of sigmoid and high rectum. Methods sixty-two gerontal patients with obstructive carcinoma of sigmoid and high rectum from April 2008 to April 2013 were randomly divided into loop ileostomy group and Hartmann group.The operation time , length of stay and postoperative complications in the two groups were analyzed. Results The time of absolute diet in loop ileostomy group was shorter than that in Hartmann group in Stage I operation (P < 0.05). The time of operation, absolute diet, and length of stay in loop ileostomy group were shorter than that in Hartmann group in Stage II operation (P < 0.05). Conclusion The application of loop iloostomy in gerontal patients with obstructive carcinoma of sigmoid and high rectum was safe and effective.

Effect of Hepatocyte Growth Factor/Scatter Factor on Apoptosis of Colorectal Cancer Cells Induced with Curcumin / 中国普外基础与临床杂志

Objective To explore the effect of hepatocyte growth factor/scatter factor (HGF/SF) on apoptosis of colorectal cancer cells induced with curcumin. Methods MTT assay was used to evaluate the cytotoxicity of curcumin to colorectal cancer cells. Flow cytometry was used to detect the anti-apoptosis effect of HGF. Results Flow cytometry showed only 64 ?g/ml curcumin could play the proliferation-inhibiting role in Caco-2 cells leading to their apoptosis; at the same time, different concentrations of HGF could antagonize this inhibitory effect resulting in the decrease of apoptosis, but HGF worked without a concentration-dependent manner. The study on MAPK pathway showed that the protective effect of HGF on the apoptosis of Caco-2 cells was not influenced by inhibiting p42/p44 MAPK and p38 MAPK pathway. Conclusion HGF/SF antagonizes the apoptosis of Caco-2 cells induced with curcumin, but MAPK signaling pathway might not participate in this process.

Heavy load exercise induced dysfunction of immunity and neuroendocrine responses in rats.

To determine whether immunity and neuroendocrine system is altered by different loads of exercise training in rats, eight-week-old male Sprague-Dawley rats were randomly assigned to one of the three groups: 1) cage control group (CCG); 2) moderate load training (MLT) (swimming at the intensity of 1.4 m/sec water flowing for 60 min per day); 3) heavy load training (HLT) (swimming at the intensity of 1.8 m/sec water flowing for 120 min per day). MLT and HLT rats were assigned to swim for 6 days per week for total of 6 weeks. All rats were sacrificed 36 h after their last training session. Splenocytes were pooled for assay of cell proliferation and neuropeptide contents in the hypothalamus, hypophysis and plasma were determined by radioimmunoassay while glucocorticoid specific binding in intact thymus was measured by radioligand binding assay. All rats were weighed weekly. The results showed that after 6-week training, rat splenocyte proliferation in response to Con A and LPS decreased in HLT rats compared with MLT and CCG rats. In addition, the contents of beta-endorphin, dynorphin A, arginine vasopressin and oxytocin in the hypothalamus, hypophysis and plasma were altered by HLT, as shown by increased plasma concentration of glucocorticoids and decreased glucocorticoids specific binding in intact thymus compared with MLT and CCG. Furthermore, a decreased body mass in HLT rats has been observed. The body mass of HLT rats was significantly lower than that in CCG and MLT rats at the end of the swimming training period. These data suggest that 6-week heavy load training induces the dysfunction of immunity and neuroendocrine responses, which might be one of the underlying mechanisms of immune dysfunction in overtraining.

Inhibitory Effects of Ethanol Extracts Obtained from Five Traditional Chinese Medicines on Experimental Hypermelanosis Induced by UVB Using a Brownish Guinea Pig Model / 中华皮肤科杂志

Objective To investigate the inhibitory effects of traditional Chinese medicine (TCM) on hypermelanosis and provide experimental evidence for treating skin pigmentary disorders. Methods Five TCMs with strong inhibitory effects on tyrosinase activity were tested. The changes of the number and morphology of melanocytes induced by UVB were observed in the experimental hypermelanosis model (brownish guinea pig). Results Decreased melanocytes and melanin granules were found with the treatment of Poria cocos, Polyporus umbellatus, Cornus officinalis Sieb. et Zucc., Atractylodes macrocephala Koidz., Astoagalus complanatus R. Br. ex Bge. Conclusion There are inhibitory effects on hypermelanosis induced by UVB with the treatment of Poria cocs, Polyporus umbellatus, Cornus officinalis Sieb.et Zucc., Atractylodes macrocephala Koidz., Astoagalus complanatus R. Br.ex Bge.

The function of HGF/SF in the proliferation of colorectal cancer cells / 中国普通外科杂志

Objective To study the function of hepatocyte growth factor/scatter factor(HGF/SF) in the (proliferation) of colorectal cancer cells.Methods The expression of c-met,the receptor of HGF,was(detected) in Caco-2 and Colo320 cell lines by Western blot.The activation of p42/p44MAPK and p38MAPK induced by HGF in these two cell lines was observed.Observation of the effect of the inhibitor of p42/p44MAPK(PD98059),p38MAPK(SB203580) on the inhibition of HGF-induced proliferation of Caco-2 and Colo320 cells were made by Using -TdR,MTT assay.Results(1)Both cell lines(expressed) the c-met.(2)HGF activated p42/p44MAPK and p38MAPK,and 20ng/ml HGF treated cells showed maximum activity in both to be within 10min.(p42/p44MAPK,2.28?0.01;p38MAPK,2.25?0.01).(3)HGF was found to significantly increase thymidine incorporation(P