A mouse model for acute hepatitis B virus (HBV) infection was established by using the hydrodynamical injection of mouse tail vein, in which the immunocompetent BALB/c mice were hydrodynamically injected with a competent replication plasmid pAAV-HBV1.2 having 1.2 fold over-length of HBV DNA. On day 1, 2, 4, 6 and 8 after injection, the levels of HBsAg, HBeAg and HBV DNA in blood serum were detected by using ELISA and fluorogenic quantitative PCR assay (FQ-PCR). And on day 8. HBsAg and HBeAg in liver tissue were assayed by immunohistochemical staining. It was found that HBsAg in blood serum could be detected on day 1 after infection in 14 of 16 mice (85.7%) injected with pAVV-HBV1.2 by using ELISA assay and the peak levels of HBsAg and HBeAg were attained during the first day after injection and then it dropped down gradually up to day 8 following injection. The titer of HBV DNA in blood serum attained its peak on day 2 and maintained a high level later on. On day 8 after injection, its titer was 1.9×10~4 copies/mL. The percentage of HBcAg-positive hepatocytes and HBsAg-positive hepatocytes in liver tissues were 5% and 2% respectively. Thus, by using the hydrodynamic injection with the competent replication plasmid, a mouse model for acute HBV infection is successively developed.

Enthusiasm is an intrinsic motive that drives the ethical and moral aspects of nursing practice.Love is the source of enthusiasm.An enthusiastic nurse loves life,work and patients.Loving patients is essential to inflame passion for patient care.A desire for and pursuits of benevolence is the key to enthusiasm.Only the element of benevolence integrated into enthusiasm can nurses arouse a sense of compassion,responsibility and honour,which is then advanced to the realm of humanitarian-Heal the Wounded,Rescue the Dying.The essence of enthusiasm is sincerity,that is,providing patients wholehearted care rather than hypocritical act.Caring for patients is based on goodwill and altruism.Enthusiasm is also the driving force for nurses' creativity.It inspires a positive life,pushes to pursue self-actualization and propels continuous creation.

Objective To investigate the value of PCT in the diagnosis of severe infection in children.Methods The clinical data of patients with infectious diseases treated in our hospital from December 2013 to February 2016 were retrospectively analyzed.According to the degree of infection,the patients were divided into local infection group and severe infection group,At the same time,the clinical data of 50 children with non infectious diseases were selected as control group.The differences of serum PCT and cytokine levels were observed between the three groups,at the same time,according to the prognosis of patients with severe infection group were divided into improvement group and deterioration group,The differences of serum PCT and inflammatory cytokines levels in the patients with severe infection in the improvement group and the worsening group,Analysis of the correlation between PCT levels and serum inflammatory cytokines levels in children with severe.Results There were significant differences in the levels of PCT,IL-18,IL-6 and hs-CRP between the three groups,which were from high to low: severe infection group,local infection group and control group; Deterioration of PCT,IL-18,IL-6,hs-CRP and levels were higher than the improvement group(t=-10.099,-8.949,-10.827,-2.088,P<0.05); The level of PCT were positively correlated with IL-18,IL-6 and hs-CRP levels in children with severe infection(r=0.385,0.412,0.408,P=0.012,0.008,0.017).Conclusion PCT has a good diagnostic value in children with severe infection,and it is closely related with the level of inflammatory cytokines in children with severe infection.

Objective The modified rapid one-step extraction (ROSE) method was employed in the Mycoplasma pneumoniae (Mp)genomic DNA extraction,in order to establish an easy DNA extraction method,which is used in Mp Loop Mediated Isothermal Amplification(LAMP) and suitable for high throughput without any expensive instruments.Methods In line with the character of Mp,a one-step Mp acid extraction method was established on the basis of ROSE.The studies had been carried out on comparing the modified ROSE DNA extraction with the classical methods of Boiling,CTAB,Alkali solution and reagent kit in extraction quality,quantity,reagent kinds,operation steps and extraction time.The extracted DNA of the Mp culture solution by these five methods had been used in LAMP to test the extraction efficiency.20 cases of Throat swab specimens from children with mycoplasma pneumonia were selected from January to March of 2016,and the throat swab specimens were extracted by the improved ROSE method then applied to LAMP detection system.Results Purity,comparison of ROSE method and other four methods were P＜0.05,the difference was statistically significant and ROSE was slightly better than the alkaline lysis method and boiling method,but lower than the CTAB method and kit method;Yield,results were compared with ROSE method and other four methods:there was no significant difference in q=0.95 (P＞0.05) compared with ROSE method and boiling method.The ROSE method was significantly different from the other three methods (P＜0.05),so the yield of ROSE method was higher than that of alkaline lysis method,CTAB method and kit method.Instrument and equipment,extracting procedure,extracting time and quantity ROSE,Boiling and Alkali solution were obvious ly better than CTAB and reagent kit.The extracted DNA by these five methods could be used in LAMP reaction,and the results showed no significant difference.Conclusion This study established an original one-step DNA genomic extraction method of Mp,which had been successfully applied in the Mp LAMP withoutany expensive instruments.Offer a groundbreaking method detecting Mp for the hospitals at all levels including the primary medical organizations.

Objective:To develop a Q-TRAP LC-MS/MS method for the content determination of militarine, protocatechuic acid and caffeic acid in Bletilla Striata. Methods:A Supelco Discovery C-18 (150 mm × 2. 1 mm, 3 μm) column was used and the mobile phase was acetonitrile-water containing 0. 1% formic acid (v/v) with gradient elution. The flow rate was 0. 25 ml·min-1. Bletilla Striata contrast medicinal materials and Bletilla Striata samples were detected by AB Sciex 4000 Q-MRM scan mode. Results: The content of militarine, protocatechuic acid and caffeic acid in the control herb of Bletilla Striata was 1. 167 5, 0. 062 6 mg·g-1 and 0. 001 0 mg·g-1 , respectively, and that in Bletilla Striata samples was 0. 708 8, 0. 001 1 mg·g-1 and 0. 000 4 mg·g-1 , respec-tively(n=3). Conclusion:The method is simple, accurate and reproducible. It can be used to quantitatively analyze militarine, pro-tocatechuic acid and caffeic acid in Bletilla Striata.

The effects of fenofibrate on plasminogen activator inhibitor-1 (PAI-1) expression in human umbilical endothelial cell-derived transformed cell line--ECV 304 cells were investigated. ECV 304 cells were incubated with different concentrations of fenofibrate (0, 10, 50, 100 micromol/L) for 24 h. PAI-1 mRNA and protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively. PAI-1 antigenic content of endothelial cells was measured by using ELISA. Fenofibrate could inhibit the PAI-1 mRNA and protein expression and reduce PAI-1 antigenic content dependently. After treatment with fenofibrate (10 micromol/L), the expression levels of PAI-1 mRNA and protein were 0.65 +/- 0.05 and 0.96 +/- 0.11 respectively, significantly lower than in the control group (0.78 +/- 0.03 and 1.21 +/- 0.15, respectively, P<0.05). PAI-1 antigenic contents (24.52 +/- 8.39) in ECV304 cells treated with 10 micromol/L fenofibrate were significantly lower than those in the control group (6.98 +/- 5.12, P<0.05). It was concluded that fenofibrate inhibited the expression of PAI-1 mRNA in ECV304 cells, and reduce the protein expression and the antigenic content of PAI-1, suggesting that fenofibrate may have an antiatherosclerotic effect on endothelial cells by PAI-1 pathway.

Objective To investigate the status of depression with anxiety symptoms, and analyze the influencing factors of anxiety symptoms from demographic data and social psychological factors. Methods Hamilton depression rat?ing scale (HAMD), Hamilton anxiety rating scale (HAMA), Eysenck personality questionnaire (EPQ), life event scale (LES), trait coping style questionnaire (TCSQ) and social support scale (SSS) were used to evaluate 729 patients with de?pression. According to HAMA scores, patients were divided into non anxiety symptoms group (HAMA<7) and anxiety symptoms group (HAMA>14). Social psychological factors were compared between two groups, and the influencing fac?tors of anxiety symptoms were analyzed. Results The incidence of anxiety symptoms in depression was 58.85% (429/729), and 119 cases (16.32%) were certainly without anxiety symptoms. Compared with the group without anxiety symp?toms, the anxiety symptoms group had higher scores on neuroticism, psychoticism, negative life events and negative cop?ing style (P<0.001), but lower scores on introversion and extroversion (P=0.010). Degree of depression (OR=9.255, 95%CI:4.726~18.127), neuroticism (OR=1.595, 95%CI:1.197~2.125), negative life events (OR=1.009, 95%CI:1.001~1.017) and negative coping style (OR=1.046, 95%CI:1.013~1.080) were the risk factors of anxiety symptoms (P<0.05). Conclu?sion The incidence of anxiety symptoms in patients with depression is high. Patients with higher degree of depression and typical neurotic personality experiencing more negative life events and those with tendency to adopt negative coping style are more susceptible to anxiety symptoms.

Objective To explore the effect of different nasal feedings in enteral nutrition for elderly patients with severe brain hemorrhage. Methods Eighty elderly patients with severe brain hemorrhage were assigned into the observation group and control group according to the odd and even numbers at admission with 40 in each group. The control group was treated with graded injection nasal feeding, and the observation group was treated with continuous infusion nasal feeding. The nutrition indicators and enteral nutrition complications were compared between the two groups. Result The nutrition indicators in the observation group were better than those in the control group and the rate of enteral nutrition complications in the observation group was lower than that in the control group (P<0.05). Conclusion Continuous infusion nasal feeding can improve the nutritional status of elderly patients with severe cerebral hemorrhage and effectively reduce the incidence of complications of enteral nutrition. So it is worthy of clinical application.

Objective To evaluate the efficacy and safety of sorafenib in the treatment of Chi-nese patients with metastatic renal cell carcinoma. Methods This muhicenter phase Ⅱ clinical trial was performed from May 2006 to December 2006. Sixty-two patients with metastatic renal cell carci-noma not suitable for curative treatment were enrolled. All patients received oral sorafenib as single a-gent at the dose of 400 mg twice a day until disease progression or intolerable toxicities occurred. Re-salts Partial responses were recorded as best response in 11 patients, while complete remission was found in 1 patient and stable diseases were found in another 35 patients. According to the intents-to-treatment population, the overall response rate was 19.4% (12/62), and the disease control rate was 77.4%(48/62). The median progression free survival time was 9.6 months with 1-year progression-free survival rate of 41.9%. However, the median survival time had not reached due to the short fol-low-up. The most frequent adverse events included alopecia (66.1%), diarrhea (62.9%), hand-foot syndrome (58.1%), anorexia (40.3%), rash(37.1%), fatigue (37.1%), hypertension (35.5%), hoarseness(32.3%), joint pain (25.8%), hypophosphatemia (21.0%), fever (19.4%), nausea (19.4%), abnormal transeaminase( 11.3% ), elevated total bilirubicin( 16.1% ), leucopenia( 12.9% ), bleeding under nail(16.1%), and gum bleeding(11.3%). Grade 3 adverse events included hand-foot syndrome (16.1%), hypertension (12.9%), diarrhea(6.5%), hypophosphatemia (4.8%), joint pain (3.2%), and leucopenia(3.2%). Conclusions Sorafenib has prominent anti-tumor activity in Chi-nese metastatic renal cell cancer patients with most adverse events being grade 1 or 2. More attention should be paid to hypertension and cardio-cerebral vascular events during the application of sorafenib.

Objective To establish normally immortalized porcine hepatocyte lines by ectopic expression of simian virus 40 large T (SV40LT) antigen and the human telomerase reverse transcriptase(hTERT). Methods Primary porcine Hepatoeyte cells were transfeeted with recombinant retrovirus containing SV40LT or hTERT respectively. Subsequently drug resistant cell clones were screened and expanded for further studies. Immortalized porcine hepatocyte was confirmed by examination. Results The morphological phenotype of the transfected cells was similar to the primary porcine hepatocyte. One clone, HepLP, has been maintained in cultue for half year, and expanded by more than 60 passages. SV40 LT and hTERT could be detected in transfected porcine hepatocyte. Pig albumin mRNA was also detected by RT-PCR. No tumor formation occurred when HepLP cells were injected into Balb/c nude mice. Conclusions The immortalized, nontumorigenic, porcine hepatoeytes maintained the properties of porcine primary hepatocytes such as the albumin secretion. This generation of immortalized porcine hepatocyte may be helpful for bioartifical liver support system, hepatocytes transplantation, drug/toxicological studies, and liver biologic studies.