Objective:To investigate the differences of gene expression and signal transduction pathways in large conductance calcium-activated potassium channels(BKCa) gene knockout rats and analyze the role of BKCa gene in pancreas.Methods:Three adult female BKCa knockout SD rats (BKCa knockout group) were donated by Professor Wang Wei from Department of Pathology and Physiology of Basic Medical College of Capital Medical University, and three wild type adult femal SD rats were used as wide-type group. The whole pancreas was resected and RNA was extracted. RNA transcriptome sequencing (RNA-seq) technology was used for sequencing and DESeq2 differentiation analysis software was used for screening differentially expressed genes between two groups, and the gene ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis were performed. The key genes were validated by RT-PCR.Results:18 258 genes were detected by sequencing in the 2 groups. There were statistically significant differences in the expression of 348 genes screened by DESeq2, 200 of which were highly expressed in the pancreas of BKCa knockout group, and 148 of which were low-expressed. 214 differentially expressed genes enrichments were found in GO database, including 25 involved in biological process, 18 in cell components and 14 molecular functions. All 348 differentially expressed genes were found in KEGG database, 15 of which were significantly enriched in PI3K/Akt signaling pathways. RT-PCR results showed that the expression of key genes Hsp90ab1, Hsp90aa1, Foxo3a and Col1a2 in the BKCa knockout group was significantly higher than that in wide type group ( P<0.0001), while Thbs1, Pik3r1 and Ppp genes were not significantly different. Conclusions:Differentially expressed genes and related important regulatory signaling pathways were screened out between BKCa knockout SD rats and wild-type SD rats at the transcriptional level, and PI3K/Akt pathway was found to be the most enriched, providing an important clue for predicting the function of BKCa in the pancreas.

Human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) might be developed as a novel anti-tumor drug due to its selective cytotoxicity in tumor cells. The predicted Macaca mulatta TRAIL (mmTRAIL) is highly homologous to hTRAIL in nucleotide acid as well as amino acid sequence, suggesting that mmTRAIL might induce apoptosis of human cancer cells. However, the cytotoxicity of mmTRAIL in human cancer cells has not been investigated. In this paper, it is reported that the gene encoding mmTRAIL has been cloned by using reverse-transcriptase polymerase chain reaction (RT-PCR) from monkey peripheral blood mononuclear cells (PBMCs) in our laboratory. Subsequently, an expression plasmid was constructed by inserting mmTRAIL gene into pQE30 plasmid. After induction by addition of Isopropyl β-D-1-Thiogalactopyranoside (IPTG), mmTRAIL was expressed. MmTRAIL was recovered from supernatant of sonicated bacteria by Ni-NTA agarose affinity chromatography. SDS-PAGE and gel filtration chromatography demonstrated that mmTRAIL forms trimer in solution. In vitro assays indicated that mmTRAIL was cytotoxic to human COLO205 tumor cells but not to normal cells at low concentration of nanomole. In addition, antitumor effect of mmTRAIL was evaluated in mice bearing human COLO205 tumor xenografts. Intratumorally injected mmTRAIL significantly inhibited growth of tumor grafts. These results suggested that mmTRAIL was valuable as candidate drug for cancer-targeted therapy.
Animals ; Antineoplastic Agents ; Apoptosis ; Cell Line, Tumor ; Cloning, Molecular ; Humans ; Leukocytes, Mononuclear ; Macaca mulatta ; Mice ; Plasmids ; TNF-Related Apoptosis-Inducing Ligand ; genetics ; metabolism ; Xenograft Model Antitumor Assays

The present article aimed at importance of self-efficacy assessment in behavioral change ,self-management capaci-ty ,empowerment capacity and active response in patients with diabetes mellitus (DM) ,introduced research progress of dia-betes self-efficacy assessment scale in China and overseas ,and influencing factors affecting self-efficacy of DM patients , such as baseline data ,well-awareness of blood glucose control ,method and content of health education and social support etc .,in order to promote research and development of Chinese self-efficacy assessment tool .

[ ABSTRACT] AIM: To investigate the role of anterior thalamic nucleus in trigeminal nerve electrostimulation ( TNS)-induced effects on seizures and hippocampal damage.METHODS: The rats were intraperitoneally injected with pilocarpine to induce chronic epilepsy, and then received sham treatment, TNS treatment and TNS treatment after stereotac-tic lesion to the anterior thalamic nucleus.The TNS treatment lasted for 1 month in each group.Another injection of pilo-carpine was conducted to induce seizures, and the severity and duration of seizures were quantitatively evaluated.TUNEL, Fluoro-Jade B ( FJB) and Nissl staining were applied to determine in situ apoptosis, neuronal degeneration and neuron loss in the hippocampal CA1 area respectively.RESULTS:Compared with TNS group, TNS treatment after stereotactic lesion to the anterior thalamic nucleus significantly increased the severity and duration of seizures (P<0.05), and the numbers of TUNEL positive cells, FJB positive cells and lost neurons in the hippocampal CA1 area (P<0.01).CONCLUSION:Anterior thalamic nucleus plays a role in TNS-induced reduction of seizures and hippocampal damage.The mechanisms might be due to the chronically activation of the cortex through anterior thalamic nucleus pathway induced by TNS, resulting in the down-regulation of neuronal excitatory susceptibility.

ObjectiveTo investigate and analyze the current primary quality management situation in TCM clinical studies.Methods One sub-center unit from 39 projects of “Prevention and Treatment of Difficult and Complicated Disease of TCM” plan of the“11th Five-year National Key Technology R&D Program” was chosen. Executive condition of the primary quality control of each unit was under field investigation, and quality control quantitative evaluation indexes were used to conduct quantitative evaluation.Results Results were shown as the forms of statistical description and radar map. The average and median scores of primary quality management were 5.05 and 5.2, respectively. The best and preferable proportions of 39 units were 25.64% and 28.21%. Other 46.15% of these units performed poor. The problem mainly manifested on insufficient frequency and bad normalization of primary quality management, such as comprehensiveness and integrity of inspection contents, cases, problem recordings and problems timely feedback.ConclusionAlthough the primary quality management is widely conducted in TCM clinical studies, there are still some problems to be settled and improved.

The neuropeptide orexins, including orexin A and orexin B, are located exclusively in hypothalamus OX1R and OX2R are receptors of orexin. OX1R is selective to orexin A while OX2R is not. The distributions of orexin neurons in human brain are the same as those of rats. Orexin was closely related with sleep and awakening behavior besides regulating the feeding behavior. Orexin-containing terminals are also found in areas modulating sleep and awakening such as the locus ceruleus, tuberomammillary nucleus, dorsal raphe, and the basal forebrain. In those areas, there are also lots of OX1R and OX2R and their precursors. Orexin neurons are extremely active during rapid wave sleep and awakening. If hypothalamus in some areas is destroyed and orexin neurons are reduced, sleep-awakening dysfunction will appear (for instance: narcolepsy).

5-Flucytosine (5-FC) could be changed to 5-fluorouracil (5-FU) by cytosine deaminase (CD), the latter is able to kill cancer cells. However, there is no efficient method to deliver the CD gene into the tumor cells, which hampers the application of the suicide gene system. In this experiment, for the first time, the NDV has been utilized as a vector to deliver the CD gene into the cancer cells, the virus can infect the cancer cells specifically, replicate and assemble, while the cytosine deaminase is expressed. Then the CD converts the prodrug 5-FC into 5-FU to achieve the purpose of inhibiting tumor. Firstly, the whole genome of E. coli JM109 was extracted, and the CD gene was obtained by cloning method. Then the CD and IRES-EGFP were ligated into the pEE12.4 expression vector to become a recombinant pEE12.4IE-CD eukaryotic expression plasmid. The human liver cancer cells were transfected with the plasmid. The cells were treated with different concentrations of 5-FC, MTT method was used to determine the killing effect of CD/5-FC system on the human liver cancer cells. The cell deaths were 18.07%, 42.98% and 62.20% respectively when the concentrations of prodrug were at 10, 20 and 30 mmol x L(-1). In 5-FC acute toxicity experiment, Kunming mice were injected with different concentrations of 5-FC at intervals of 1:0.5. The LD50 of 5-FC through iv injection was determined by improved Karber's method, the LD50 was 507 mg x kg(-1) and the 95% confidence limit was 374-695 mg x kg(-1). According to the maximum LD0 dose of the LD50, the maximum safe dose was 200 mg x kg(-1). Body weight and clinic symptoms of the experimental animals were observed. These results laid the foundation to verify the antitumor effect and safety of CD/5-FC system in animal models. The CD gene was ligated into the NDV (rClone30) carrier, then the tumor-bearing animal was established to perform the tumor inhibiting experiment. The result showed that the recombinant rClone30-CD/5-FC system has a high antitumor activity in vivo. To summarize, CD gene has been cloned and its bioactivity has been confirmed in the mammalian cells. It is the first time in this study to utilize the recombinant NDV to deliver the CD gene into the tumor cells; our result proves the rClone30-CD/5-FC system is a potential method for cancer therapy.

OBJECTIVE:To analyze intervention effect of Drug Rational Usage Guidelines System(DRUGS)on antibacterial agents use in department of obstetrics and gynecology in our hospital. METHODS:The application of antibacterial agents in depart-ment of obstetrics and gynecology in our hospital during Jan.-May(before intervention)and Jun.-Nov. 2012(after intervention)were extracted in respects of drug name,preoperative medication duration,perioperative additional condition,postoperative drug with-drawal time,drug combination,usage and dosage,average hospitalization stay,hospitalization cost. The intervention effects were analyzed. RESULTS:After intervention,the type of antibacterial agents were more in line with national regulations;the proportion of type Ⅰ incision surgery without antibacterial agents increased from 57.8% to 75.2%;the prophylactic application of antibacteri-al agents in type Ⅱ incision surgery within 0.5-2 h increased from 80.2% to 97.0%. The rate of reasonable antibacterial selection, drug combination,usage and dosage increased from 76.9%,64.9%,71.3% to 89.3%,84.6%,90.2%,respectively. The average hospitalization stay and antibacterial cost per capita decreased significantly. There was statistical significance among above indica-tors before and after intervention(P<0.05 or P<0.01). CONCLUSIONS:DRUGS effectively change irrational use of antibacterial agents in department of obstetrics and gynecology,which provide a new method for the management of antibacterial agents.

Objective To evaluate the role of glycogen synthase kinase-3 beta (GSK-3β) in spinal cord ischemia/reperfusion (I/R) injury in rats.Methods Forty-eight male Sprague-Dawley rats,aged 3 months,weighing 250-300 g,were randomly divided into 3 groups (n =16 each) using a random number table:sham operation group (group S),group I/R and I/R+ GSK-3β inhibitor LiCl group (group LiCl).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.Spinal cord ischemia was induced by 45 min occlusion of the abdominal aorta followed by reperfusion.In I/R and LiCl groups,normal saline 5 ml and LiCl 15 mg/kg were injected,respectively,via the caudal vein at 30 min before ischemia.The animals were sacrificed at 48 h of reperfusion and the lumbar segment (L4-6) of spinal cords was removed for microscopic examination and for determination of neuronal apoptosis in the anterior horn of the spinal cord (by TUNEL),and the expression of interleukin-6 (IL-6),IL-8 and IL-10 was detected (by immunohistochemistry).The apoptosis rate was calculated.Results Compared with group S,the apoptosis rate was significantly increased,IL-6 and IL-8 expression was upregulated,and IL-10 expression was down-regulated in I/R and LiCl groups.Compared with group I/R,the apoptosis rate was significantly decreased,IL-6 and IL-8 expression was down-regulated,IL-10 expression was up regulated,and the pathological damage was attenuated in LiCl group.Conclusion Activated GSK-3β is involved in the development of spinal cord I/R injury possibly by promoting synthesis and release of inflammatory factors in rats.

A Co2+ ion-mediated formaldehyde imprinted polymer ( MIP) was prepared by coordination polymerization method in present work and its surface structure characterized by using IR spectrum and scanning electron microscope ( SEM). Inversed phase gas chromatography ( IGC) technique using this formaldehyde imprinted polymer as stationary phase was utilized to investigate on the retention selectivity, isotherm adsorption and adsorption thermodynamics for this imprint material toward the template and its structural analogue. Also, the ability of this polymer in the removal of formaldehyde from room atmosphere was explored. Results indicated that the capacity of the template on the molecularly imprinted polymers (MIPs) column was much higher than that of aldehyde and the lower column temperature and flow rate of carrier gas was beneficial for the selective retention of imprint material toward the template molecule, possessing a higher capacity factor of 61. 1 for the template and a higher separation factor of 10. 66 for this imprint polymer toward formaldehyde and aldehyde under the optimized chromatographic conditions ( column temperature: 363 K;flow rate of carrier gase: 7. 0 mL/ min; injection volume: 3. 0 μL). An approximate linear adsorption isotherm for the template and a BET Ⅲ one for the analogue on the MIPs was observed. In addition, this molecularly imprinted polymer was shown with higher capability in the removal of formaldehyde from room atmosphere.