Objective:To evaluate and compare the identification of several DNA barcoding candidate sequences on Illicium difengpi and its fake I. jiadifengpi. Method:Samples from different origins of I. difengpi and I. jiadifengpi, were collect extraction of total DNA,nuclear gene ITS2 sequence,chloroplast rbcL,matK gene sequence were selected for PCR amplification,product purification and sequencing,and CondonCode Aligner V3.7.1 was used to proofread stitching. Result:PCR amplification and sequencing of rbcL sequences of I. difengpi and I. jiadifengpi were not satisfactory. It is assumed that their rbcL sequences were too long with slow evolution,which was unsuitable for interbreeding. The success rate of matK sequencing of I. difengpi and I. jiadifengpi was 0 and 76.8%,which may be because primer standards were different for matK sequences of different groups. The results of PCR amplification and sequencing of ITS2 on I. difengpi and I. jiadifengpi were successful,with the success rate of sequencing was 89.3% and 91.2%. Analysis sequencing results, the total length of ITS2 sequences was 268 bases,and there were 2 variation sites of I. difengpi. The total length of ITS2 sequences was 430 bases,and there were 4 or 3 variation sites of I. jiadifengpi. It shows that ITS2 sequences of I. difengpi and I. jiadifengpi were short and has obvious variability and can be amplify,that ITS2 sequence was better than rbcL and matK sequence in molecular identification of I. difengpi and I. jiadifengpi. Conclusion:DNA barcoding based on ITS2 sequence was a powerful and efficient tool for identification of I. difengpi and its fake I. jiadifengpi.

Objective To construct adenovirus vector containing firefly luciferase reporter gene (AdLuc) and infect bone marrow mesenchymal stem cells (BMSC),then to take bioluminescence imaging in vitro and in vivo for identification.Methods The luciferase gene was amplified with PCR from psiCHECK-2 plasmid and cloned into the adenoviral shuttle vector (pShuttle-CMV).It was confirmed by Nhe Ⅰ/Xba Ⅰ digestion and sequencing.PShuttle-CMV-Luc and backbone vector (pAdeno) were homologous recombined.Then the recombinant plasmid was packaged in HEK293 cells and the virus titer was detected.The BMSC were infected by the recombinant adenovirus.The bioluminescence imaging in vitro was performed to determine the best multiplicity of infection (MOI),and the relationship between bioluminescence intensity and MOI was analyzed by curve fitting regression analysis.Viability was evaluated via Trypan blue staining.The transfected BMSC (l× 106) were implanted into the muscles of forelimb of SD rats,and then tracked by bioluminescence imaging in vivo.Cell viability was compared using two-way repeated measures analysis of variance between groups.Results Enzyme digestion and sequence analysis indicated that Ad-Luc was successfully constructed.The virus titer was 1 × 1010 plaque forming unit (PFU)/ml.The bioluminescence detection in vitro showed that Ad-Luc could infect BMSC high efficiently to express luciferase and the best MOI was 50.The bioluminescence intensity enhanced with increase of MOI (R2 =0.98).No statistically significant difference was found in cell viability between transfected and untransfected BMSC at 1,3,5,7 d.The cell survival rates were (92.5±2.3)％ vs (94.1±1.8)％,(91.4±0.9)％ vs (92.7±2.0)％,(92.1±1.6)％ vs (93.3± 2.4) ％,(91.9 ± 1.5) ％ vs (93.0 ± 3.1) ％,respectively (F =4.38,P ＞ 0.05).The bioluminescence imaging in vivo showed that BMSC survived 1,3,7 d after implantation.However,bioluminescence signal decreased gradually over time.Conclusion It is feasible to apply the optical reporter gene imaging for tracing transplanted stem cells in vitro and in vivo due to the effective transformation of luciferase reporter gene into BMSC by adenovirus vector.

To test the hypothesis that dermal fibroblasts (DF) are an important source of cytokines which elicit major changes in hepatic synthesis of acute phase reactants(APRs). Metkods: Condi-tioned medium(CM) from human DF challenged with bacterial lipopolysaccharide (I,PS) was collected andIL-10 and IL-6 levels were measured- The ability of DF conditioned medium(fLPS) and dexamethasone(Dex) to mediate an hepatic acute phase response was tested on rat hepatoma H4 cells- Various concentra-tions and combinations of CM (?LPS), recombinant IL-6 (rhIL-6) and Dex were tested for their abilitiesto stimulate albumin, Q,-acid glycoprotein (AGP), a,-antitrypsin (AT) and transferrin mRNA synthesis.Results: LPS stimulated IL-6 production by DF and Dex inhibited this producti0n. IL-6 and CM+LPS in-hibited the production of albumin mRNA,while the expression of AT and AGP 0ccurred 0nly with CM+LPS+Dex. However,IL-6 alone had an inhibitory effect 0n albumin and transferrin mRNA producti0n.Dex maximized the effects 0f lL-6 and CM, and was essential f0r AGP gene expression. C0nclusion: (l)LPS-treated human DF can secrete IL-6, but not IL-101 (2)DF may als0 pr0duce other cyt0kines whichmodulate hepatic pr0tein synthesis during the acute phase response l (3) Dex inhibits IL-6 production byDF, but enhances its ability to stimulate the acute phase response.

Objective To analyze the multi-slice computed tomography (MSCT) enhanced findings in the patients with primary small intestinal stromal tumor(SIST),and to probe the relationship between the imaging findings and the pathologic risk in order to improve the diagnostic accuracy.Methods Thirty patients with primary SIST confirmed by surgical pathology were enrolled in this study.Characterization and compassion of the clinical manifestations and MSCT enhanced findings were carried out between the pathologic low-and high-risk groups.Furthermore,the relationship was analyzed between the enhanced findings and the pathologic risk.Results Among all 30 patients with primary SIST,the lesion was located at duodenum in 5 patients (16.7％),at jejunum in 16 (53.3％),and at ileum in 9 (30％).14 patients were classified in the low risk group with the lesion with the average length of (3.8±0.9) cm,and other 16 in the high-risk group with lesion with the average length of (7.0 ± 1.4) cm.There were no statistical differences between the low-and high-risk groups in CT value in plain and venous phase,and in added value in arterial,venous and delayed phases.However,the significantly differences were observed in CT value in arterial and delayed phases between two groups (P＜0.05).Conclusion MSCT may effectively evaluate the pathologic risk of primary SIST.There are significant differences of the enhanced findings between the low and high-risk groups,which can provide important apreoperative classification for the therapy.

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Objective To evaluate the effectiveness and safety of autologous granular fat graft applied for penile lengthening and augmentation.Methods After all the superficial ligment and 1/3-2/ 3 part of the deep suspensory-ligament had been cut off for penile lengthening,local pedicaled fasciaadipose flap was designed to fill the depression,the pre-centrifuged autologous granular fat was injected into the space beneath Buck's fascia for penile augmentation.Normal length,pulling penis length,diameter,circumference and complications were evaluated.Results 34 cases were performed and followed up for 3-18 months,both ideal length and diameter increase of penis were achieved.The differences of nomal length,pulling-length,the diameter and circumference were (2.8±0.1) cm,(2.1±0.2) cm,(0.9 ± 0.1) cm,(2.8 ± 0.1) cm,respectively.The common complications included poor wound healing in 4 cases,preputial edema and subcutaneous scleroma in 8 cases for 3 months.Conclusions Autologous granular fat graft for penile augmentation during the lengthening surgery is a reliable and effective method and easy procedure.Detail processing can decrease the complications.

Objective To learn about the antibiotic resistance status of methicillin resistant coagulase negative staphylococcus(MRCNS),and to investigate the distribution and resistant feature of different staphylococcal chromosomal cassette mec(SCCmec) genotypes of children in Anhui,so as to guide clinical medication.Methods Resistance phenotype screening was conducted in coagulase negative staphylococcus,which were isolated from clinical strains in children in Anhui from 2010 to 2014 each year in September.MecA gene was detected by using PCR method in order to collect MRCNS.Minimal inhibitory concentrations (MIC) of 16 antibiotics were determined by adopting agar dilution method.Vacomycin-resistant strains were identified with population analysis and the Brain Heart Infusion vancomycin screen agar dilution method recommended by Clinical and Laboratory Standards Institute in 2013.Van gene and SCCmec types were detected by using PCR method.Results A total of 148 MRCNS strains were detected through the resistance phenotype screening and the detection of mecA gene.There were methicillin resistant staphylococcus epidermidis,methicillin resistant staphylococcus haemolyticus,methicillin resistant staphylococcus hominis,and other kinds of MRCNS,and the proportions of them were 44.59％ (66/148 cases),25.68％ (38/148 cases),19.59％ (29/148 cases) and 10.14％ (15/148 cases),respectively.The analysis of antibiotic resistance showed the antimicrobial resistant rates of MRCNS to Penicillin,Cefoperazone,Cefotaxime,Ceftriaxone,lmipenem and Meropenem were all 100％,to Erythromycin and Azithromycin,Ciprofloxacin,Clindamycin,Gentamicin,Lewofloxacin,Rifampincin,Chloramphenicol,Teicoplanin and Vancomycin were 92.57％,97.98％,83.78％,79.05％,43.24％,35.81％,24.32％,8.78％,2.03％ and 0.68％,respectively.There was 1 heterogeneous Vancomycin-resistant strain,which was resistant to both Vancomycin and Teicoplanin (with MIC 32.00 mg/L and 64.00 mg/L).No vanA,vanB,vanC1 or vanC2/3 gene was detected from heterogeneous Vancomycin-resistant strain by PCR.Ⅰ to Ⅴ SCCmec genotypes were detected from 148 MRCNS strains,and the major SCCmec type was SCCmec type Ⅲ,which was followed by hybrid type.Three subtypes of SCCmec type Ⅳ were identified,including Ⅳa,Ⅳc and Ⅳd.There were 148 MRCNS strains that showed different resistant phenotypes to various antibiotics.Conclusions The MRCNS strains of children in Anhui province showed multiple resistance to antibiotics.It should be on alert when heterogeneous Vaneomycin-resistant strain appeared.There were several different SCCmec types among several kinds of MRCNS,and SCCmec Ⅲ genotype was the major epidemic isolate.There was no significant correlation between the different resistance rates of non-β-lactamase antibiotics and SCCmec genotypes in MRCNS.

Objective To discuss the clinical value of high sensitivity C‐reactive protein(hs‐CRP) ,fibrinogen(Fib) and D‐dimer (D‐D) measurement for patients with acute myocardial infarction(AMI) before and after the treatment with the anticoagulation and thrombolysis therapy .Methods 110 patients with AMI were recruited in the study and the plasma hs‐CRP ,Fib ,D‐D and myocardi‐al damage markers were measured before and after the treatment .Results 66 of the 110 patients′plasma hs‐CRP ,Fib ,D‐D concen‐trations elevated(higher than the threshold) before treatment and after treatment within 24 h ,while 44 patients′plasma hs‐CRP , Fib concentrations increased ,but D‐D didn′t .Conclusion The measurement of hs‐CRP is helpful for the diagnosis and treatment of AMI .Hs‐CRP is another good myocardial injury marker ,and the plasma hs‐CRP concentration after treatment for 24 -48 h could reflect the severity and prognosis of AMI better than after treatment within 12 h .Fib decreases relatively slowly after the treat‐ment ,so it cannot be used for curative effect observation for AMI patients;D‐D concentration dosen′t have the determined negative predictive value for the diagnosis of AMI ,so it cannot be used as screening out indicator for AMI ,but D‐D concentration can be used as therapeutic effect monitoring indicator for AMI patients with D‐D positive .

OBJECTIVE:To select a best therapeutic scheme for diabetes among four kinds of oral hypoglycemic drugs:repaglinide(Novonorm),glibenclamide,gliclazide and glipizide.METHODS:According to the principle of pharmacoeconomics,using cost-effectiveness analysis(CEA),assessment and survey of the influencing factors(price,dosage)were carried out,at the same time,sensitive analysis was performed.RESULTS:The best therapeutic scheme was Novonorm and the rest were gliclazide,glipizide and glibenclamide in sequence.CONCLUSION:The result of sensitive analysis is in keeping with that of CEA,which indicates the credibility of CEA.