Objective To study the impact of ethanol in nucleic acid extraction products on quantitative real-time PCR(qPCR), and to provide ethanol concentration limits for the extraction quality of more sensitive nucleic acid detection methods.Methods A potassium dichromate oxidation method was developed to detect the ethanol residues in the products extracted by two commercialized magnetic bead nucleic acid extraction kits, testing whether the ethanol residues impact qPCR. Then different concentrations of ethanol solutions were mixed with qPCR reagents that had been dissolved in simulated extraction products and freeze-dried(hereinafter referred to as freeze-dried reagents) for amplification. The sensitivity changes were observed, and the Ct values, fluorescence gain intensity and amplification efficiency were statistically analyzed.Results The extraction products of the two types of reagent kits contained 3%-9% of residual ethanol, both of which caused an impact on the amplification, potentially leading to a decrease in sensitivity. For human cytomegalovirus(HCMV), when ethanol concentration reached 4. 38%, the fluorescence gain intensity showed significant difference(P < 0. 001), and the amplification efficiency showed significant difference. For respiratory syncytial virus(RSV), there was a significant difference in fluorescence gain intensity when ethanol concentration reached 2. 63%(P = 0. 004). When the concentration of ethanol reached 3. 50%, the difference of Ct values was significant(P = 0. 004), and the amplification efficiency was significantly different.Conclusion Ethanol can affect the sensitivity, Ct values, fluorescence gain, and the amplification efficiency of qPCR. In order to detect nucleic acid with higher sensitivity, it is suggested that the residue of ethanol in the extracted products should not exceed 3. 50%.

Objective To analysis the occurrence and risk factors of ventilator-associated pneumonia(VAP)in intensive care unit in order to direct the clinical work.Method The clinical data of 158 cases receiving mechanical ventilation in ICU were retrospectively analyzed to investigate the incidence of VAP and risk factors.Result Mechanical ventilation and intubation of nasogastric tubes were the main risk factors of VAP(P<0.01).Conclusion The nursing measures such as earlier extubating, keeping the oropharynx clean,decreasing the risk of colonization,strengthening disinfection and air purification,paying attention to the tracheotomy nursing and enhancing nursing procedure skills are important for VAP prevention.

ObjectiveTo analyze the clinical characteristics of atypical acute myocardial infarction,improve the diagnosis of myocardial infarction. MethodsTo analyze the main clinical characteristics of the patients with atypical acute myocardial infarction in our hospital in the past 3 years ,which compared to the typical acute myocardial infarction. ResultsAtypical acute myocardial infarction includes atypical symptom and atypical ECG, atypical symptoms is more prevail. Compared with typical acute myocardial infarction, the women, elderly people and hypertension patients is more common, fewer anterior myocardial infarction and poorer prognosis ( P ＜ 0.05 ). ConclusionTo improve the understanding of atypical acute myocardial infarction,was benefit of reducing the misdiagnosis and mistreatment of acute myocardial infarction.

Objective To investigate the clinical manifestations and diagnosis and treatment of aortic dissection(AD).Methods The clinical data of 20 patients with aortic dissection were analyzed retrospectively.Results 20 patients were diagnosed by CT, MRI and colour ultrasonography (5,5,10 cases, respectively). According to DeBakey typing, this group of patients was composed of type I(11 cases, 55%), type II(2 cases, 10%)and type III (7 cases, 35%). All patients were treated with internal medicine ,average time in hospital stay was(29 8?25 5)days. The patients' conditions were improved in 19 person-times (76%). 4 patients died (20%) and 2 patients abandoned treatment (8%).Conclusions The accurate diagnosis as early as possible and active therapy was a key of improvment of prognosis in patients with aortic dissection.

To explore the clinical features,diagnosis and treatment of Langerhans cell neoplasms.The data of 6 patients with Langerhans cell neoplasms was analyzed retrospectively.There were Langerhans cell histiocytosis (LCH,n =5) and Langerhans cell sarcoma ( LCS,n =1 ).Among 5 LCH patients,the involved organs included waist,back,scalp,inguinal lymph nodes,parotid gland and left femur.And the symptoms were fever,splenohepatomegalia and destruction of bone.Three patients received surgery,1 had radiotherapy and 1 was discharged without treatment.The follow-up period was 34 - 138 months.Two patients had no relapse,the contra-lateral parotid gland became involved later in another and several organs were involved in other patients.One patient was lost to follow-up.Left occiput of another LCS was involved and both surgery and radiotherapy were administered.Over a follow-up period of 41 months,there was no relapse.Either surgery or radiotherapy is indicated for this rare disease when a single organ is involved.And chemotherapy is administered for the involvement of multiple organs.The patients should be treated individually.

Objective To investigate the effect of continuing nursing on elderly patients with high risk of pressure sores. Methods High-risk elderly patients with pressure sores healed in our hospital in January 2014 to December 2014 were divided into control group and observation group with 130 cases in each group by random number table. The control group received routine follow-up service, while the observation group were intervened by continuing nursing care. To observe the differences before and after the interventions in both groups on the patient's quality of life and the incidence of pressure sores. Results There was statistically significance on mental function scores (t=12.393, P=0.000), social function score (t=8.459, P=0.000) and overall quality of life score (t=12.750, P=0.000) between the two groups after the interventions. Meanwhile, there was no statistically significance on material life score (t =-1.913, P=0.057) and physical function score (t=-0.328, P=0.744). The difference of the re-occurrence of pressure sores in the two groups was statistically significant (χ2=35.96, P=0.000), either. Conclusions Continuing nursing intervention can obviously reduce the risk the re-occurrence of pressure sores on high-risk elderly patients, in particular to improve the quality of life aspects of mental function and social function in patients, meanwhile with no obvious improvement in that of material life and body functions.

The activation of p21 kinase 2 (PAK2) has a certain value in cancer research, since its participation in a series of intracellular biological activities. There is a relatively little research on its relationship with the development of tumors. PAK2 can be activated by a variety of upstream signals, especially small G protein Rho family of Rac and Cdc42, and it participates in a variety of important signaling pathways and cell function regulation. Abnormal expression of PAK 2 has been reported in various tumors, which involves in cytoskeletal remodeling, cell motility, apoptosis, signal transduction, gene transcription, translation and angiogenesis. Therefore, PAK2 plays an important role in tumor occurrence and development. The study on PAK2 and the targeting therapy for anti-PAK2 provides a new sight on the treatment and prevention of tumor.

Objective To explore preliminarily the role of the E2 subunit of pymvate dehydrogenase (PDC-E2) modified by xenobiotics (e.g.2-octynic acid,2-OA) in the pathogenesis of primary biliary cirrhosis (PBC).Methods Patients of PBC (102 cases),primary sclerosing cholangitis (PSC,34 cases) and healthy controls (HC,50 cases) were selected.The anti-PDC-E2,anti-2-OA and anti-lipoic acid (LA) antibody in the peripheral blood of the 3 groups were tested by enzyme linked immunosorbent assay (ELISA).By inhibitive ELISA (iELISA),30 of the 102 PBC patients with anti-PDC-E2 antibody but without anti-2-OA antibody were selected to detect whether there was any new epitope on the PEC-E2 conjugated with 2-OA.The chi-square test and Fisher exact test were taken to analyze the enumeration data.The two-tailed unpaired t test with Welch's correction was used to compare the measurement data.Spearman rank correlation analysis was also employed for proper test.Results The positive rate of anti-PDC-E2,anti-LA and anti-2-OA antibody in PBC patients was 94.1％(96/102),73.5％(73/102) and 53.9％(55/102) respectively,all of which were statistically significantly higher than those in healthy controls group but were of no significant difference between PSC and healthy controls group.There was no significant relevance between the levels of Anti-LA and anti-2-OA antibody in the PBC group (r=-0.065,P=0.520).The iELISA results showed that the antibody,which only identified the epitopes on 2-OA-PDC-E2 induced by the 2-OA conjugation with PDC-E2,existed in 40％(12/30) of the PBC patients,and more interestingly,this antibody was predominantly appeared in PBC patients at their early clinical stage.Conclusion There are anti-LA antibody and anti-2-OA antibody in PBC patients,which have shown no significant association with each other.It is very likely that new antigenic conformational epitopes on PDC-E2 modified by 2-OA would emerge,which might led to the immune response in the individuals who are susceptible to PBC,and thus contribute for the breaking of PDC-E2 immune tolerance,and PBC occurrence finally.

Evidence-based medicine(EBM) emphasizes that the clinical practice should be based on the combination of the best available clinical evidence,medical experiences and patient desire to cope with the demand of medical decision-making.Guidelines for good clinical(research) practice,conduct of more trials as multicentre trials,and the cochran collaboration may all help apply the best research evidence to clinical practice.China is challenged by variety of health problems.The implementation of EBM will improve the decision-making and health care services and protect the doctors and patients rights.

Objective To investigate the effect of cyanidin-3-glucoside extracted from Chinese bayberry on the proliferation. migration and invasion ability of the gastric cancer cell lines MGC803 and SGC7901 in vitro, and explore the possible mechanism of the preventive effects of C3G on tumor metastasis.Methods After treatment by C3G, the growth inhibiting of C3G on MGC803 and SGC7901 was determined by MTF assay, cell migration and invasion ability was evaluated with transwell chamber. Expression of Matrix metalloproteinase 2( MMP-2 )mRNA and protion on cells were analyzed by RT-PCR and Western blot.Results C3G significantly inhibited the proliferation of MGC803 and SGC7901 cells in a concentration and time-dependent manner as measured by MTT method ( P ＜0. 01 ), and the IC50 were: MGC803:24 h IC50 =6. 27 μg/ml;SGC7901:24 h IC50 = 5.42 μg/ml. After the cells were treated with C3G, the migration and invasion ability of MGC803 and SGC7901 cells decreased significantly ( P ＜ 0. 01 ) the number of invasive cells in 24 hours of the negative control MGC803, SGC7901 group was ( 207 ± 9 ) and ( 115 ± 9 ),respectively, while in C3G 10 μg/ml group the number of invasive cells decreased to( 24 ± 5 ) , ( 14 ± 6). In addition, the expression of MMP-2 mRNA and protion decreased abviously ( P ＜ 0. 01 ), all that was in a concentration-dependent manner. Conclusions In vitro, C3G has a concentration- and time-dependent growth inhibiting effect on MGC803 and SGC7901 cells, and may prevent metastasis by affecting migration and invasion ability of tumor cells. This action may be mediated by down-regulation of MMP-2mRNA and protein.