Objective To study the genomic DNA extraction from Rhizoma Coptidus and optimization of RAPD reaction system. Methods Different methods, i.e. phenol method, CTAB method, low pH extraction medium with high salt, were used to genomic DNA extract from Rhizoma Coptidus. The DNA samples obtained by the above methods were tested by agarose gel electrophoresis and ultraviolet spectrometer. Results CTAB Method was considered to be an optimal technique. Based on the genomic DNA extracted by CTAB method, a reaction system suitable for Rhizoma Coptidus was established, that is, 25 ?L amplification reactions system containing 1?PCR buffer, 2 mmol/L Mg 2+, 100—150 ?mol/L dNTP, 20 ng primer, 40 ng template DNA, and 1 U Taq DNA polymerase. Conclusion CTAB Method and RAPD reaction system can be used to RAPD analysis in Rhizoma Coptidus.

Objective To investigate the protective effect of the extract of ginkgo biloba on myocardial tissue in aged rat.Methods The rats aged 20 months were given the extract of ginkgo biloba (EGB) and ALT-711 respectively by garage for 16 weeks.The aged controls and adult rats were infused with the same volume saline.The superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), malonyt diadehyde (MDA) in blood samples and advanced glycation end products (AGEs) in the myocardial tissue were measured .The myocardial histopathological changes under electron microscope and the mitochondrial DNA (mtDNA) deletion rate in myocardial cells were observed.Results Compared with the adult rats, the content of AGEs in myocardial cells in aged rats was significantly increased [(33.5±1.3)AU/mgHYP us.(18.1±1.2)AU/mgHYP, t= 7.18,P<0.05] and the levels of SOD and GSH-PX in blood samples were decreased [(138.4±3.8) U/mlvs.(227.7±13.8)U/ml, (1283.8±28.8) U/ml vs.(2114.1 ±135.9)U/ml, t=-19.59, -18.79;both P<0.01].The MDA level in the serum and mtDNA deletion rate in aged rats were higher than in adult rats[(6.7±0.6) mmol/ml vs.(4.1±1.0) mmol/ml, (0.18054±0.0718) % vs.( 0.0060±0.0001)%, t=7.18,6.98;both P<0.05].Compared with the aged controls, the content of AGEs in myocardial cells, the level of MDA and mtDNA deletion rate were significantly decreased in EGB and ALT-711 treatment adults (all P<0.05).The SOD and GSH-PX in blood samples were increased in EGB and ALT-711 treatment adults (all P<0.05).Conclusions Nonenzymatic glycation may play an important role in myocardial aging, which may be amplified by oxidative stress.EGB and ALT-711 may have the same anti-aging effects by inhibiting nonenzymatic glycation and oxidative stress.

Objective To investigate the role of cancer stem cells in radiation resistance of esophageal cancer and its molecular mechanism, and to provide a theoretical basis for radiotherapy for esophageal cancer.Methods Esophageal cancer cell line TE1 was treated with 8 Gy of radiation. Esophageal cancer cell line with resistance to radiation, TE1-res, was established and screened.Cell counting was used to evaluate cell proliferation.Flow cytometry was used to determine the expression of CD44 (high) CD24(-) CD133(+) and apoptosis in cells.The colony formation assay was used to determine the colony-forming rate and cell survival curve.Bisulfite sequencing PCR was used to determine the methylation status of cancer suppressor genes.Comparison of the data was made by group t test or analysis of variance. Results Compared with TE1 cells, TE1-res cells had significantly enhanced proliferation, a significantly higher proportion of CD44( high) CD24(-) CD133(+) cells, and significantly enhanced resistance to apoptosis (mean value 20.84×105 vs.4.46×105/day, P=0.008;(38.0±2.9)%vs.(10.1±1.3)%, P=0.001;mean value 33.23% vs.10.50%, P=0.003 ) .After treatment with 8 Gy of radiation, TE1-res cells had significantly higher colony-forming rate and D0 value than TE1 cells ((14.3±2.6)%vs.(0.9±0.3)%, P=0.011;3.28 vs.2.19 Gy, P=0.125 ) .Moreover, the promoter methylation in cancer suppressor genes including SPINT2, CDKN1B, DKK1, TP53, and PPP2R1B was significantly enhanced in TE1-res cells than in TE1 cells ((89.7±4.9)%vs.(5.0±0.5)%, P=0.001;(92.3±4.7)%vs.(10.4±0.7)%, P=0.001;(90.7±3.7)%vs.(7.9±0.4)%, P=0.001;(83.4±5.7)%vs.(17.2±1.2)%, P=0.002;(90.2±
6.7)%vs.(4.4±1.2)%, P=0.002).Conclusions Cancer stem cells play an important role in radiation resistance of esophageal cancer. The resistance to radiation is closely associated with promoter hypermethylation in cancer suppressor genes including SPINT2, CDKN1B, DKK1, TP53, and PPP2R1B.

Objective To establish a radioresistant esophageal squamous cancer cell line,and to identify the radioresistant genes and mechanisms.Methods The radioresistant KYSE410-res cell line was established by repeated exposure of cell line KYSE410 to radiation.The proliferation and apoptosis of esophageal squamous cancer cells were evaluated before and after radiation.The changes in gene expression of the esophageal squamous cancer cells after radiation were determined by gene microarray and analyzed by group t test.The genes with significant difference in expression after radiation were validated.Results The KYSE410-res cells had significantly enhanced proliferation and anti-apoptosis than the KYSE410 cells (all P＜0.05).The result of gene microarray showed that compared with the KYSE410 cells,the KYSE410-res cells had the expression of 463 and 251 genes upregulated and downregulated by no less than 4 folds,respectively.Those genes with different expression levels after radiation were mainly responsible for cell proliferation,adhesion,signal transduction,angiogenesis,reactive oxygen metabolism,cell damage repair,and the MAPK/ERK signaling pathway.OAS2 and UBD were key proteins in the network.In the KYSE410-res cells,the expression of HLA-DQBI,MMP1,NCAM1,ZNF521,GPC6,SELENBP1,LCN15,and TFPI-2 genes measured by real-time PCR was consistent with that measured by gene microarray.Conclusions Abnormal activation of the MAPK/ERK signaling pathway,upregulated expression of OAS2 and UBD,downregulated expression of TFPI-2,and upregulated expression of MMPs may play a role in radioresistance of esophageal cancer cells.

Objective To determine the vitamin D status in children with sepsis/severe sepsis in pediatric intensive care unit (PICU) in order to explor the association between vitamin D status and clinical outcomes,in turn to provide evidence for optimizing nutrition support.Methods It was a prospective,observational,multi-center study,carried out in patients with sepsis/severe sepsis from March 1,2013,to March 30,2014,in the PICUs of three tertiary-care children's hospitals.Total serum 25-hydroxy vitamin D [25 (OH) D] was measured by using an enzyme-linked immunosorbent assay at admission.The association of vitamin D status at admission with length of PICU length of stay,total hospital stay,in-hospital mortality,28-days mortality and costs were analyzed.Results A total of 194 patients includng 117 boys (60.3％)and 77 girls (39.7％) were enrolled.There were 96 patients with sepsis and 98 with severe sepsis.The mortality on discharge and 28 days were 6.7％ and 24.2％ respectively.The median vitamin D level was 9.79 ng/mL (5.32,18.46) at admission.Of them 77.8％ (151/194) had vitamin D deficiency and 50.5％ (98/194) had severe vitamin D deficiency.Patients with severe vitamin D deficiency,had higher mortality on discharge (P =0.011).Vitamin D status had no significant correlations with 28 days mortality,length of PICU stay,total hospital stay and costs.Conclusions More than three-quarters (77.8％) of children with sepsis/severe sepsis in PICUs had Vitamin D deficiency.Patients with severe vitamin D deficiency at admission had higher risk of mortality at discharge.

Acetamides ; metabolism ; toxicity

The paper introduces application and development of medical information technologies and elaborates the overall reform of medical science undergraduate curriculum made by Oregon Health and Science University including the reform method and the curriculum provision after reform.It points out the enlightenment for medical informatics curriculum reform of China from the perspectives of intensif-ying utilization of computing devices, valuing interaction with patients, enhancing combination with clinical practices, etc.

Objective: : To observe whether cytokines rIL 2,TNF ?,I FN ? and anti CD3/anti glioma bispecific antibody(BsAb) can work coordinately, and to investigate how to further enhance cytotoxicity of T lymphocyte against human glioma cells by BsAb. Methods: There were 12 groups,contr ast method were used to analyze the effect of cytokines rIL 2,TNF ?,IFN ? to cytoxicity directed by BsAb by single and combined experiments. Cytotoxicity was assayed by standard 18 h 3H TdR incorporation release. Resul ts: rIL 2,TNF ?,IFN ? and BsAb could cooperatively enhance the cy totoxicity of effect cells( P

Objective To evaluate endoscopic ultrasonography (EUS) for diagnosis of large gastric folds. Methods EUS was performed in 66 patients with possible large gastric folds which could not be diagnosed by conventional gastroscopy. The characteristics of EUS findings were analyzed, and the EUS results were compared with pathological findings to evaluate the sensitivity, specificity and accuracy of EUS. Results The diagnostic sensitivity, specificity and accuracy of EUS for diffused infiltrated gastric cancer were 92.3%(24/26), 95.1% (39/41) and 95.5% (63/66), respectively, and those for gastric lymphoma were 92. 3% ( 12/13), 96. 1% (49/51) and 92. 4% (61/66), respectively. The accuracy in differential diagnosis of benign and malignant disease was 93.9% (62/66). There were statistical differences between benign and malignant large gastric folds in characteristics of EUS findings, including width of gastric wall, enlargement of muscularis propria and preservation of gastric layer ( P ＜ 0. 05 ). Conclusion EUS is of high accuracy for diagnosis of large gastric folds, especially for diffused infiltrated gastric cancer and gastric lymphoma. Such features as width of gastric wall, the enlargement of muscularis propria and the preservation of gastric layer are important features for differential diagnosis of benign and malignant lesions.

Objective To investigate the effect of once yearly zoledronic acid of 5 mg on postmenopausal women with osteoporosis of different causes. MethodsFrom October 2009 to December 2009,a total of 89 postmenopausal women with osteoporosis were enrolled and assigned into 2 groups.There were 45 cases of primary postmenopausal osteoporosis,including 27 cases of fresh fracture,in group A.They were aged from 47 to 83 years,with an average of 63.7 years.There were 44 cases of secondary postmenopausal osteoporosis,including 28 cases of fresh fracture,in group B.All patients were given a.single 30-minute intravenous injection of zoledronic acid (5 mg),supplemented by 1,25-dihydroxyvitamin D of 0.25 μg and calcium of 600 mg with VitD125 IU daily.At pre-intervention and 12 months after intervention respectively,bone mineral density (BMD) was measured by dual-X-ray absorptiometry (DXA) at the lumbar spine and hip,and a balance test(Sunlight Tetrax- Ⅱ) was performed to evaluate the risk of falling.Intervention compliance of the patients and adverse events related to zoledronic acid infusion were observed. Results All cases of fresh fracture healed well at 3-month follow-up.At 12 months,43 subjects in group A and 42 subjects in group B completed the follow-up.In group A,BMD increased by 5.8％ at the lumbar spine,by 2.9％ at the femoral neck,by 5.2％ at the Words area,by 5.3％ at the greater trochanter and by 3.9％ at the total hip while the risk of falling decreased by 26.1％; in group B,BMD increased by by 3.4％ at the lumbar spine,by 2.1％ at the femoral neck,by 3.2％ at the Words area,by 3.0％ at the greater trochanter and by 2.5％ at the total hip while the risk of falling decreased by 21.8％.The differences between pre-intervention and post-intervention were significant in both groups ( P ＜ 0.05).No intolerable adverse events occurred in both groups except that one new fracture happened in each group but responded to conservative treatment.ConclusionA once-yearly infusion of zoledronic acid of 5 mg is a convenient and effective therapy for treatment of osteoporosis in postmenopausal women.