Objective:To establish a nicotine-treatment and-withdrawal rat model and to evaluate its characteristics and application through analyzing 3 parameters.Methods:Male Sprague-Dawley rats,aged 10-11 weeks old,were randomly divided into normal saline group(subcutaneous injection of saline [0.5 ml/kg] for 6 weeks),nicotine-treated group(injection with nicotine [0.5 ml/kg,3 mg/(kg?d)] for 6 weeks),and nicotine-withdrawn group(injection with nicotine [0.5 ml/kg,3 mg/(kg?d)] for 3 weeks and followed by saline injection for additional 3 weeks).Body weight,food intake,and water intake of animals were recorded during the treatment in 3 groups.The model was evaluated through analyzing body weight,serum parameters and adipose tissue weights.Results:The body weight of rats,as well as the serum levels of triglyceride and insulin,were all decreased after nicotine treatment;the weights of subcutaneous fat,visceral fat and periaortic fat were also decreased.The above indicators increased after withdrawal of nicotine.Conclusion:The established model can be used to study multiple pharmacological effects of nicotine;it can also be used for smoking and smoking cessation related studies.

Objective To investigate relationship between CD133 expression and progression of hepatocellular carcinoma (HCC) by detecting expressions of CD133. Methods 65 cases with HCC were enrolled and their HCC tissues and peripheral blood samples were taken. Expression of CD133 in HCC tissue was detected with immunohistochemical method and the percentages of CD45- CD133+ cells in peripheral blood were assayed by the method of flow cytometry (FCM), and 10 healthy persons were chosen as controls. Results In HCC patients, the positive expression rate of CD133 in HCC tissues was 6.4 % and the percentage of CD45- CD133+ cells in peripheral blood was 2.0 %. The expression rate of CD133 was not found in normal liver tissues and peripheral blood (P <0.01). The amount of CD133+ cells was significantly related to portal vein tumor embolus (P<0.01), tumor amicula integrality (P<0.01), pathology grade (P<0.01) and TNM stage (P <0.01), but not related to tumor diameter(P0.05) and AFP value (P0.05). It showed a positive correlation between HCC tissues and peripheral blood (r=0.36, P <0.01). Conclusion The percentage of CD133+ cell in HCC tissues and peripheral blood from HCC patient is greatly correlated to the progression of HCC. It is useful to exam the CD133 expression in predicting the diagnosis and prognosis evaluation.

Objective To discuss the effectiveness between arthroscope cleaning combined with exercise therapy and exercise therapy in senile osteoarthritis patients.Methods Two hundred and ten senile osteoarthritis patients were divided into operation group and non-operation group by random digits table method with 105 cases each.Patients in operation group received the arthroscope cleaning and exercise therapy,and patients in non-operation group received the exercise therapy alone.The visual analogue score (VAS) and international knee documentation committee knee evaluation form (IKDC) score before treatment and 1,6 and 12 months after treatment were compared.Results A total of 210 patients received the analysis of result without drop-out.There were no statistical differences in VAS and IKDC scores before treatment and 6 and 12 months after treatment between operation group and non-operation group (P ＞ 0.05).The VAS and IKDC scores 1,6,12 months after treatment were significantly improved compared with those before treatment in operation group,and there were statistical differences (P ＜ 0.05).The VAS and IKDC scores 6,12 months after treatment were significantly improved compared with those before treatment in non-operation group,and there were statistical differences (P ＜ 0.05).According to the IKDC score,in operation group 15 patients were excellent,61 patients were fine,19 patients were general and 10 patients were bad after treatment,and good effectiveness rate was 72.38％ (76/105).In non-operation group,14 patients were excellent,59 patients were fine,20 patients were general and 12 patients were bad after treatment,and good effectiveness rate was 69.52％ (73/105).There was no statistical difference in the good effectiveness rate between 2 groups (P ＞ 0.05).Conclusions Exercise therapy with guidance can significantly improve the function of senile knee joint osteoarthritis,and ease joint pain symptoms.Arthroscope cleaning combined with exercise therapy can relieve the joint acute inflammatory symptoms,but it has no significant clinical effectiveness in long-term treatment of osteoarthritis.

Objective To investigate the expressions of hepatorna stem cell surface marker CD133 CD90 in tissues of hepatocellular carcinoma (HCC) and evaluate their related clinical significances. Method The expressions of CD133 CD90 were detected by immunohistochemical method in HCC tissues of 93 patients, and normal liver tissues of 10 cases. Results Among 93 cases with HCC, the positive expression of CD133 were found in 71 cases (76.3%), and CD90 positive expression in 64 cases (68.8%), and the percentage of positive cells were (6.4±3.3)% and (4.3±3.9)% respectively. No positive expression of CD133 and CD90 was found in normal liver tissues (P<0.01). CD133, CD90 expressions in the HCC tissues of TNM stage Ⅲ-Ⅳ [(8.1±3.7)%,(5.7±4.2)%] were higher than those of TNM stage Ⅰ - Ⅱ [(4.1±2.3)%,(2.3±1.9)%] (P<0.01). Spearman correlation analysis indicated that the expressions of CD133 and CD90 were up-regulated as the pathohistology grades increased (P<0.05). Positive correlation was observed between CD133 and CD90 expression (r=0.402, P<0.01). Conclusions CD133, CD90 positive cells exist in HCC tissues, their expressions positively relate to the TNM stage and the pathohistology grades for HCC patients.

Objective To investigate the coverage of a recombinant protein vaccine based on pneumococcal surface protein A (PspA) from both family 1 and family 2.Methods One hundred and fifty-nine Streptococcus pneumoniae strains, including 47 invasive strains, were isolated from children in Nanjing Children′s Hospital.Cell lysates were prepared and reacted with three antibodies recognizing PspA -RX1, PspA-3296 and PspA-5668 for PspA typing by ELISA .Results Among 47 invasive isolates of 9 different serotypes, 10.7%were PspA family 1 and 89.3%were PspA family 2.Among all of 159 clinical isolates, 10.1% were identified as PspA family 1, 88.0%were family 2, while 1.9%of strains could not be typed by ELISA and PCR assays .None of strains belonged to PspA family 3.Conclusion The recombinant pro-tein vaccine based on PspA from both family 1 and family 2 has a broad coverage among clinical isolates and is potentially protective against both invasive and non-invasive pneumococcal diseases .

Objective To investigate T-bet mRNA expression on peripheral blood mononuclear cells (PBMCs) and its relations with allergen specific IgE (SIgE), eosinophile cationic protein (ECP) levels, and allergic symptom in patients with allergic rhinitis (AR). Methods The allergen, SIgE, and ECP in serum of patients with AR were detected by Unicap CAP system. Blood samples were taken from 15 healthy controls and 35 house dust mite allergic patients. PBMC was isolated by density gradient centrifugation and one part of them was cultured with mite allergen at a concentration of 50 μg/mL. PBMC was subjected to analysis of T-bet mRNA expression using semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Results The ratio of T-bet to β-actin mRNA levels was 0.418 ± 0. 101 in patients of AR and 0.706 ± 0.091 in healthy controls and the difference was significant (P ＜ 0.01). The expression intensity of T-bet mRNA was not related to varying severity of allergic symptom and ECP levels ( r = - 0.227, - 0.033, P ＞ 0.05). However, there was an inverse correlation between expression intensity of T-bet mRNA and SIgE concentration (r = -0.375, P ＜ 0.05). There was a positive correlation between SIgE and allergic symptom scores ( r = 0.426, P ＜ 0.05). After that PBMC was stimulated by mite allergen, the expression intensity of T-bet mRNA, ECP, and SIgE changed very little ( P ＞ 0.05). Conclusion Down-regulated expression of T-bet mRNA in mite-AR patients is not related to serum ECP and symptom scores but one of important links in the mechanism of imbalance of Th1/Th2 in the occurrence of AR. Specific allergen has no effect on T-bet mRNA, ECP, and SIgE of children and adults with AR in vitro. The level of SIgE objectively and directly indicates the severity of allergic symptom, but T-bet did not. T-bet may be one of indirect factors which affect the level of IgE.

Objective To analyze the positive report time,distribution and antimicrobial resistance of pathogens isolated from blood culture at a hospital,so as to provide laboratory basis for prevention,contro1 ,and rational antimicrobialuse for bloodstream infection.Methods From January 2013 to January 2015,blood culture specimens of outpatients and inpatients were performed bacterial identification and antimicrobial susceptibility testing, antimicrobial resistance was analyzed.Results A total of 1 973 blood culture specimens were sent by clinical depart-ments,219 (11 .10%)of which were isolated pathogens.Most positive blood culture specimens were from depart-ment of paediatrics (n = 199 ).Isolation rates of gram-negative bacteria,gram-positive bacteria,and fungi were 44.34% (n=98),50.23% (n=111),and 5.43% (n=12)respectively;the main pathogens was coagulase-negative staphylococcus (n=53,23.98%),followed by Escherichia coli (n=39,17.65%),Staphylococcus aureus (n=23, 10.41 %),Klebsiella pneumoniae (n =15,6.79%),and Pseudomonas aeruginosa (n =13,5.88%),the average positive blood culture report time of top five pathogens was 1 -2 days.The detection rates of extended-spectrumβ-lactamase-producing Escherichia coli and Klebsiella pneumoniae accounted for 53.85% and 53.33% respectively, susceptibility of gram-negative bacilli to carbapenems was relatively high(76.92% - 100%);methicillin-resistant isolates accounted for 39.13% among Staphylococcus aureus and 64.15% among coagulase-negative staphylococ-cus,vancomycin-resistant and teicoplanin-resistant strains were not found;resistant rate of Candida glabrata to 5-fluorocytosine was 14.29%,but was susceptible to amphotericin B.Conclusion The major pathogens isolated blood culture are gram-positive bacteria,in order to reduce the emergence of drug-resistant strains,clinicians should choose antimicrobial agents according to blood culture results and antimicrobial susceptibility testing results.

Objective To study and investigate the efficacy of alteplase combined with low molecular weight heparin sodium in the treatment of patients with submaximal pulmonary embolism and its influence on related indicators, and to guide clinical medication. Methods Eighty patients with large pulmonary embolism treated in our department from August 2013 to August 2017 were randomly divided into observation group and control group, with 40 cases in each group. The control group was treated with subcutaneous injection of low molecular weight heparin sodium. In combination with oral anticoagulation with warfarin, the observation group was treated with alteplase on the basis of the control group. After 1 week of continuous treatment, the clinical efficacy was compared between the 2 groups, and pulmonary function and arterial partial pressure of oxygen (PaO2) were compared before and after the treatment. The partial pressure of carbon dioxide (PaCO2) was used to compare the levels of serum B-type natriuretic peptide (BNP) , troponin Ⅰ (cTnⅠ) , homocysteine (Hey) , Ddimer (D-D) , and bleeding before and after the treatment in both groups. Results The total effective rate was97.50％ in the observation group, which was significantly higher than that in the control group of 75.00％ (P <0.05). There was no significant difference in terms of FEV1％, FEV1/FVC, PaO2 and PaCO2 between the two groups before the treatment (P> 0.05) , but FEV1％, FEV1/FVC, PaO2 in the observation group were significantly higher than those in the control group after the treatment, and PaCO2 was significantly lower (P <0.05); There was no significant difference regarding to serum BNP, cTnl, Hey and D-D levels between the two groups before the treatment (P> 0.05) , but after the treatment, the indexes of the observation group were significantly lower than those of the control group (P < 0.05). There was no statistical difference in the incidence of bleeding between the two groups (P> 0.05). Conclusion Alteplase combined with low-molecular-weight heparin sodium in the treatment of the next large-area pulmonary embolism can improve the efficiency of treatment, lung function and the prognosis, but will not inc-rease the risk of bleeding. It could be widely used in clinical practice.

Nicotinic acetylcholine receptors (nAChRs) ,also known as neuronal nicotinic receptors ,are widely expressed throughout the central and peripheral nervous system .nAChRs play crucial roles in pain signaling .Recently ,agonists that tar-get specific nAChR subtypes have shown substantial efficacy in a wide range of acute and chronic pain models ,contributing to developing novel analgesic drugs with low drug dependence .Positive allosteric modulators offer new approaches for increasing the potency and therapeutic window of these drugs .This review summarizes the advances in nicotinic acetylcholine receptors as analgesic targets and anticipate the future directions in this field .

Objective Nicotinamide phosphoribosyltransferase (Nampt) is a new therapeutic target for ischemic stroke. The aim of this study was to investigate protective effect of liver-derived Nampt on ischemic stroke. Methods Liver-specific Nampt knockout mice were generated using the Cre/loxP system. Nampt^loxP/loxP mice were crossed with liver-specific Cre recombinase expression mice (Alb-Cre), and the progeny genotypes were identified by polymerase chain reaction. Body weight of knockout mice and control mice were measured. Nampt in liver and brain was determined by Western blot assay. Middle cerebral artery occlusion (MCAO), a classical ischemic stroke model, was generated in liver-specific Nampt knockout mice and control mice by electrocoagulation. After 24 h of modeling, neurological deficit scores of each group were evaluated and TTC staining was performed to determine the cerebral infarction volume. The level of plasma Nampt in each group was determined by ELISA. Results Liver-specific Nampt knockout mice with the genotype of Nampt^loxP/loxPAlb-Cre were successfully constructed. The hepatic Nampt expression in knockout mice was significantly decreased by 74.2% compared to control mice, while there was no significant difference in the expression of brain Nampt protein between the knockout group and the control group. Specific knockout of liver Nampt gene expression had no effect on the body weight of mice. Under normal physiological conditions, there was no significant difference in plasma Nampt levels between liver-specific Nampt knockout mice and control mice of the same gender. 24 h after MCAO modeling, there were no significant differences in neurological deficit scores, cerebral infarct volume and plasma Nampt concentration between liver-specific Nampt knockout group and control group. Conclusion Liver-specific Nampt knockout mice are successfully constructed. Liver-derived Nampt has no significant protective effects on ischemic stroke.