Objective To study the repairment of pancreas mesenchymal stem cells(MSCs) on injured pancreas of rats,and find a new source of cells for treatment of diabetes. Methods ① Pancreases were taken out of three-day-old Wistar rats under bioclean condition,and cells were obtained by V-collagenase digestion. Generations were passed by conventional culture,cells were purified by adherence screening method,cell morphous was observed by Giemsa staining. The expressions of surface marker CD34 and CD44 were determined by FCM,and compared with bone marrow MSCs(BM-MSCs),the differences in character and function were observed. ②The experimental rats were divided into two groups randomly. Pancreas ischemic necrosis model was made by deligation,then the purified pancreas MSCs were marked with DAPI and then were transplantated partially. After two weeks,the survival rate was measured and histopathological detection was performed. Results ① The cells had concordant morphous gradually with vigorous generation. There was no significant difference in morphology and surface antigen compared with BM-MSCs.② The survival rate in experimental group was 75% ,the necrotic tissue had basi-rebounded. Blue fluorescent was observed in repaired pancreas tissues. The survival rate in control group was 20%. The survival rate in experimental group was higher than that in control group (P

Objective To investigate the changes of PPARr mRNA and protein expressions in rats with ANP. Methods 36 rats were randomly divided into sham group and ANP group. The rats were sacrificed 3 h, 6 h,12 h after ANP induction, the levels of serum amylase were measured, the pancreatic pathological changes were determined and the expressions of pancreatic PPARr mRNA and protein was examined by RT- PCR and immunohistochemistry. Results In ANP group, the level of serum amylase at 6h was (7170.83± 1635.59) U/L, the scores of pathological changes were 6.67±1.03 and 13.00±2.36, which were much higher than those of sham group (P<0.01) ; the PPARr mRNA expression was 0.18±0.05, and there were no obvious differences compared with that of sham group (0.22±0.03 ) ; PPARr protein expression was 4.17 ±0.98, which was significantly higher than that of sham group (1.83±0.71, P<0.05). Conclusions Inflammatory injury resulted in increased deactivation of pancreatic acinar PPARr, meanwhile PPARr gene expression was inhibited by feedback.

Objectives To analyze comparatively the adverse reactions of Nedaplatin(NDP)and Cisplatin(DDP)in concurrent chemoradiotherapy for locally advanced nasopharyngeal carcinoma and summarize the nursing points as well.Methods From March 2012 to March 2013,112 patients with locally advanced nasopharyngeal carcinoma were randomly divided into NDP group and DDP group.Besides intensity modulated radiotherapy for both groups,NDP group were treated with intravenous drop infusion of NDP by 100 mg/m2 and the control group with intravenous drop infusion of DDP by 100 mg/m2 both for three courses of once every three weeks (e.g.day one,day 22 and day 43 during the course).The two groups were compared in terms of therapeutic effects and incidences of adverse reactions.Results The complete remission rates of the NDP group and DDP group were 87.5%and 85.7%,respectively (P>0.05).The incidences of adverse reactions like gastrointestinal reactions and radioactive mucositis in the NDP group were significantly lower than those in the DDP group(P<0.05)and the index of platelet decrease was significantly higher(P<0.05).There were no significant differences between the two groups in terms of liver and kidney dysfunction and white blood cells decrease(P>0.05). Conclusions Chemotherapy with NDP combined with radiotherapy for locally advanced nasopharyngeal carcinoma has fewer adverse effects and is easy to be accepted by patients so that their quality of life can be improved.In the application of the two kinds of chemotherapy,we should pay attention to the adverse reactions on patients in order to give pertinent care.

OBJECTIVE: To construct a cell model with simultaneous knockout of RNA methyltransferase-like( METTL) 3 and METTL14 using clustered regularly interspaced palindromic repeats( CRISPR)/CRISPR-associated protein 9( Cas9)system in human bladder cancer T24 cells,in order to provide new tools and means for further studying gene intervention in occupational cancers. METHODS: The small guide RNA( sgRNA) oligonucleotide sequences targeting METTL3 and METTL14 were designed,synthesized and constructed into the Lenti-multi-CRISPR vector. The Lenti-iCas9-neo inducible plasmids were transfected into T24 cells by lentivirus system,and the positive cells were sorted by flow cytometry. Finally,the recombinant plasmid Lenti-multi-METTL3-METTL14 was transfected into positively sorted cells and screened to obtain stable cell lines. T24-Lenti-multi-CRISPR cells and T24-KO-METTL3-METTL14 were used as control group and multigene knockout group. The Western blot was used to examine relative expression of METTL3 and METTL14 protein in the two groups. RESULTS: The targeting sgRNA was successfully inserted into the Lenti-multi-CRISPR plasmid and confirmed by sequencing. The flow cytometry was used to obtain a large amount of the positive cells. The stable cell line inducted of doxycycline with simultaneous knockout of METTL3 and METTL14 gene was obtained. The relative expression of METTL3 and METTL14 protein decreased respectively by 52. 08% and 64. 04% after 96 hours of doxycycline treatment in the multigene knockout group compared with the control group( P < 0. 01). CONCLUSION: The stable T24 bladder cancer cell with simultaneous double gene knockout of METTL3 and METTL14 was obtained for the first time,which will be helpful for providing a foundation for mechanistic study for prevention and treatment of candidate tumor targets and bladder cancer.

Stroke is an acute cerebro-vascular disease with high incidence and poor prognosis, most commonly ischemic in nature. In recent years, increasing attention has been paid to inflammatory reactions as symptoms of a stroke. However, the role of inflammation in stroke and its underlying mechanisms require exploration. In this study, we evaluated the inflammatory reactions induced by acute ischemia and found that pyroptosis occurred after acute ischemia both in vivo and in vitro, as determined by interleukin-1β, apoptosis-associated speck-like protein, and caspase-1. The early inflammation resulted in irreversible ischemic injury, indicating that it deserves thorough investigation. Meanwhile, acute ischemia decreased the Sirtuin 1 (Sirt1) protein levels, and increased the TRAF6 (TNF receptor associated factor 6) protein and reactive oxygen species (ROS) levels. In further exploration, both Sirt1 suppression and TRAF6 activation were found to contribute to this pyroptosis. Reduced Sirt1 levels were responsible for the production of ROS and increased TRAF6 protein levels after ischemic exposure. Moreover, N-acetyl-L-cysteine, an ROS scavenger, suppressed the TRAF6 accumulation induced by oxygen-glucose deprivation via suppression of ROS bursts. These phenomena indicate that Sirt1 is upstream of ROS, and ROS bursts result in increased TRAF6 levels. Further, the activation of Sirt1 during the period of ischemia reduced ischemia-induced injury after 72 h of reperfusion in mice with middle cerebral artery occlusion. In sum, these results indicate that pyroptosis-dependent machinery contributes to the neural injury during acute ischemia via the Sirt1-ROS-TRAF6 signaling pathway. We propose that inflammatory reactions occur soon after oxidative stress and are detrimental to neuronal survival; this provides a promising therapeutic target against ischemic injuries such as a stroke.

We report a complete genomic sequence of rare isolates (minor genotype) of the SARS-CoV from SARS patients in Guangdong, China, where the first few cases emerged. The most striking discovery from the isolate is an extra 29-nucleotide sequence located at the nucleotide positions between 27,863 and 27,864 (referred to the complete sequence of BJ01) within an overlapped region composed of BGI-PUP5 (BGI-postulated uncharacterized protein 5) and BGI-PUP6 upstream of the N (nucleocapsid) protein. The discovery of this minor genotype, GD-Ins29, suggests a significant genetic event and differentiates it from the previously reported genotype, the dominant form among all sequenced SARS-CoV isolates. A 17-nt segment of this extra sequence is identical to a segment of the same size in two human mRNA sequences that may interfere with viral replication and transcription in the cytosol of the infected cells. It provides a new avenue for the exploration of the virus-host interaction in viral evolution, host pathogenesis, and vaccine development.
Base Sequence ; China ; Cluster Analysis ; Gene Components ; Genetic Variation ; Genome, Viral ; Genotype ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; Sequence Analysis, DNA ; Severe Acute Respiratory Syndrome ; genetics