Objective To observe the expression of NOD-like receptor pyrin domain-3 (NLRP3) inflammasome and interleukin-1 beta (IL-1b) in pterygium and normal conjunctiva specimens,and clarify the role of NLRP3 in the development of pterygium.Methods Specimens from 20 cases of pterygium and 6 cases of normal eonjunctival were analyzed for establishing the expression of NLRP3 and IL-1b by immune-histochemistry.The level of caspase-1 and pro-caspase-1 protein were analyzed by Westernblot,gene expression of interleukin-18 (IL-18) was detected by RT-PCR.Results NLRP3 was not expressed in normal conjunctival epithelial cells,but was positive in the basal part of pterygium;IL-1was not expressed in normal conjunctival tissues,but was positive in pterygium.There was no significant difference about the expression of Procaspase-1 in normal conjunctiva and pterygium (P ＞ 0.05),However,caspase-1,also known as the active form of pro-caspase-1 expressed in pterygiun was higher than that in normal conjunctiva(P ＜ 0.05).The average level of IL-18 in 20 cases of pterygium was significantly higher than that in normal conjunctiva (P ＜ 0.05).Conclusion After NLRP3 signaling pathway activating in pterygium tissue,caspase-1,IL-1β and IL-18 are high expression abnormally,suggest that NLRP3 inflammasome and the related signaling pathways may play a role in the progression of pterygium.

Objective To establish a method for content determination of harpagide and harpagoside in Mailuoning injection by HPLC. Methods The experimental condition of HPLC method was as follows: SunfireTM C18 column (4.6 mm ×150 mm, 5 μm), with gradient elution using acetonitrile and 0.03% phosphoric acid; the detected wavelength was 210 nm, and the flow rate was 1.0mL/min.ResuIts Harpagide and harpagoside demonstrated good linear relationship in the range 0.1424~0.8544 μg/mL(r=0.9998) and 0.0732~0.4392μg/mL (r=0.9997) respectively.The average recovery rate were 98.22% and 99.27% with RSD of 1.46% and 1.42%(n=6)respectively.ConcIusion The method is simple, reliable, accurate, reproducible and stable, and it could be used in the determination of harpagide and harpagoside in Mailuoning injection.

OBJECTIVE:To investigate the dosage and consumption sum of opioid analgesic drug in Qingdao district. METH-ODS:The consumption data of drugs in 29 public hospitals at secondary or above level in Qingdao district were analyzed statistical-ly by ABC analytic methods and defined daily dose methods. RESULTS:In ABC analysis,5 kinds of class A drugs accounted for 20% of the total number of species,and the percentage of consumption sum was 77.38%;4 kinds of class B drugs accounted for 16.00% of all species numbers,and the percentage of consumption sum was 11.98%;other 16 kinds of drugs accounted for 64.00% of the total number of species,and the percentage of consumption sum was 10.64%. Oxycodone sustained-release tablets and Morphine sustained-release tablets with the highest DDDs consumed more health care costs,serial number ratio was 1.00,syn-chronization was good and conform to the actual needs of clinical work. CONCLUSIONS:The composition ratio of opioid analge-sic drug costs is consistent with the theoretical value,significant discrepancy between cost and DDDs does not appear.

OBJECTIVE: To evaluate the efficacy and toxicity of sorafenib plus Capecitabine in patients with advanced hepatocellular carcinoma (HCC). METHODS: 20 patients (treatment group) were assigned to take sorafenib 200 mg bid for 3 consecutive weeks plus capecitabine 1 500 mg? m-2?d-1 for l4 days followed by 7 days discontinuation in 3-week treatment cycle. 22 patients in the control group only received Capecitabine 1 500 mg?m-2?d-1 for l4 days followed by 7 days discontinuation in a 3-week treatment cycle. Tumor response was assessed after 2-cycle treatment using modified WHO criteria. RESULTS: In the treatment group and the control group: the median survival times were 10.9 months and 7.2 months, respectively; the median time for tumor progression was 6.8 months and 4.3 months, respectively; the overall response rates were 20.0% and 9.1% respectively; the clinical benefit rates were 70.0% and 40.9%; the ?-foetoprotein (AFP) reduction rates were 65.5% and 39.0%, respectively. The toxicities were not significant between the two groups. CONCLUSION: Sorafenib plus Capecitabine is safe and effective for advanced hepatocellular carcinoma patients.

OBJECTIVE:To discuss the influence of drug price reduction and regulation of purchase on the use of anti-infectives in our hospital.METHODS:The use of anti-infectives in5times of drug price reduction was retrospective anal?ysed.RESULTS:The proportion of the sum of money for consumption of anti-infectives in the total sum for drug consumption dropped year by year but the total DDDc of anti-infectives increased year by year.The DDDs and the total DDDc of all kinds of anti-infectives showed different characteristics.CONCLUSION:The macroscopic price reduction for anti-infectives posi?tively affects the rational use of anti-infectives,yet the effect is limited.Microcosmic regulation of purchase offers an indis?pensable effect.

Objective To construct a recombinant adenoviral vector carrying and co-expressing human inhibitor of growth 4(ING4) and human interleukin-24(IL-24) mediated by poly( A)-Promoter[Ad-ING4-poly(A)-Promoter-IL-24, referred to as Ad-ING4-IL-24] and explore its effect on the growth of HepG-2 human hepatocellular carcinoma cellsin vitro. Methods The poly(A)-Promoter(hEFl-elF4g) (Sal Ⅰ and Not Ⅰ ), ING4 ( Bgl Ⅱ and Sal Ⅰ ), and IL-24 ( Xho Ⅰ and Xba Ⅰ ) fragments were amplified by PCRusing pORF-mbcl-2α, pcDNA3.0-IL-24, and pcDNA3.0-ING4 plasmids as templates and subcloned into pAdTrack-CMV transfer vector to form pAdTrack-CMV-ING4-poly (A)-Promoter-lL-24, respectively. The pAdTrack-CMV-ING4-poly (A)-Promoter-IL-24 transfer vector linearized with Pme Ⅰ digestion and pAdEasy-1 backbone vector was further cotransformed into the bacteria BJ5183 competent cells for homologous recombination. The resultant pAdEasy-l-pAdTrack-CMV-ING4-poly ( A )-Promoter-IL-24 homologous recombinant plasmids were linearized with Pac Ⅰ digestion and transfected into the human embryonic kidney 293 (QBI-293A) cells by liposome, leading to formation of the recombinant adenoviruses Ad-ING4-IL-24co-expressing ING4 and IL-24. The Ad-ING4-IL-24 were amplified in QBI-293A cells and its titer was up to 3.5 × 109 PFU/ml. Adenovirus-mediated ING4 and IL-24 genes expression in HepG-2 cells was examined by RT-PCR and Western blot. The growth-suppressing and apoptosis-inducingg effect of Ad-ING4-IL-24 coexpressing ING4 and IL-24 on HepG-2 human hepatocellular carcinoma cells was assessed by MTT assay and FCM, respectively. Results DNA sequencing showed that the ING4, poly (A)-Promoter, and IL-24 fragments subcloned into pAdTrack-CMV plasmids were completely identical to those reported in GenBank.ING4 and IL-24 gene mediated by adenovirus could both successfully express in HepG-2 cells. Adenovirusmediated ING4 and IL-24 co-expression significantly suppressed HepG-2 hepatocellular carcinoma cell growth and induced cell apoptosis, and the effect of Ad-ING4-IL-24 group was more significant than AdING4 and Ad-IL-24 group. Conclusion The adenoviral vector co-expressing ING4 and IL-24 mediated by poly(A)-Promoter(Ad-ING4-IL-24) was successfully constructed. Ad-ING4-IL-24 had marked anti-tumor effect in suppressing HepG-2 human hepatocellular carcinoma cell growth and inducing cell apoptosis in vitro. Compared with Ad-ING4 and Ad- IL-24, Ad-ING4-IL-24 enhanced anti-tumor effect.

OBJECTIVE: To investigate the relationship of the function and gene polymorphism of insulin receptor substrate (IRS) with type 2 diabetes mellitus (T2DM), and to provide a new perspective for T2DM drug development. METHODS: Relevant literatures included in CNKI, Wanfang, VIP, PubMed, SpringerLink and other databases from Jan. 1991 to Nov. 2017 were retrieved by using "Insulin receptor substrate" "Type 2 diabetes" "Insulin resistance" "Polymorphism" as Chinese keywords, and "Insulin receptor substrate" "IRS" "Type 2 diabetes" "Insulin resistance" "Polymorphism" as English keywords. The relationship of the function and gene polymorphism of IRS family with T2DM was reviewed. RESULTS & CONCLUSIONS: A total of 328 literatures were retrieved, of which there were 38 valid literatures. At present, IRS family has six members (IRS-1 to IRS-6). The dysfunction of IRS-1 and IRS-2 will lead to insulin resistance and induce T2DM. The relationship of IRS-3 and IRS-4 with T2DM remains controversial. IRS-5 and IRS-6 were newly found and their functions are not clear. The Gly972Arg mutation of IRS-1 is positively correlated with the pathogenesis of T2DM. Gly1057Asp mutation of IRS-2 combined with obesity can induce insulin resistance, but there is controversy. The mutation types of IRS family other members include Ala94Thr, Ala512Pro and Ser892Gly mutation of IRS-1, ACC, Ala157Thr and Leu647Val mutation of IRS-2. The relationship between these types of mutation and T2DM has not yet been fully supported. Multiracial and large-scale studies are required. Some achievements have been made in the present study, but the study is not yet comprehensive. Relationship of IRS family members and their mutation sites with T2DM still needs to be further tested in the expanded population.

OBJECTIVE:To provide reference for rational selection of antibiotics against non-fermentative Gram-negative bacilli in clinic. METHODS:Etiological data of clinical isolated Pseudomonas aeruginosa(PA),Acinetobacter baumanii(AB) and Stenotrophomonas maltophilia(SM)were collected from the Affiliated Hospital of Qingdao University(called"our hospital"for short)during Jan. 2004-Dec. 2016. Drug resistance of them to commonly used antibiotics was analyzed retrospectively. RESULTS:Totally 15 587 strains of PA,7 446 strains of AB and 2 950 strains of SM were detected. Resistance rates of PA to commonly used antibiotics fluctuated but were in a decreasing tendency. Except for imipenem,resistance rates of PA to commonly used antibiotics decreased significantly,and resistance rates of PA to amikacin and gentamicin decreased to 4.60% and 7.48%, respectively. Resistance rates of AB to most commonly used antibiotics were more than 40%,but it was sensitive to tigecycline (drug resistance of 0-4.03%). Resistance rates of SM to cefoperazone sodium and sulbactam sodium increased from 3.03% in 2004 to 39.01% in 2016,but it was sensitive to sulfamethoxazole,minocycline and levofloxacin. CONCLUSIONS:Non-fermentative Gram- negative bacilli detected in our hospital are mainly PA. Resistance rate of PA to most of the antibiotics is declining;drug resistance of AB is severe;resistance rates of SM to cefoperazone sodium and sulbactam sodium show increasing tendency.Above 3 non-fermentative Gram-negative bacilli are sensitive to amikacin,tegocycline and minocycline. Clinical selection should be based on the results of drug sensitivity test.

The corona-like spikes or peplomers on the surface of the virion under electronic microscope are the most striking features of coronaviruses. The S (spike) protein is the largest structural protein, with 1,255 amino acids, in the viral genome. Its structure can be divided into three regions: a long N-terminal region in the exterior, a characteristic transmembrane (TM) region, and a short C-terminus in the interior of a virion. We detected fifteen substitutions of nucleotides by comparisons with the seventeen published SARS-CoV genome sequences, eight (53.3%) of which are non-synonymous mutations leading to amino acid alternations with predicted physiochemical changes. The possible antigenic determinants of the S protein are predicted, and the result is confirmed by ELISA (enzyme-linked immunosorbent assay) with synthesized peptides. Another profound finding is that three disulfide bonds are defined at the C-terminus with the N-terminus of the E (envelope) protein, based on the typical sequence and positions, thus establishing the structural connection with these two important structural proteins, if confirmed. Phylogenetic analysis reveals several conserved regions that might be potent drug targets.
Amino Acid Sequence ; Antigens, Viral ; immunology ; Base Composition ; Computational Biology ; Enzyme-Linked Immunosorbent Assay ; Membrane Glycoproteins ; genetics ; Molecular Sequence Data ; Mutation ; genetics ; Phylogeny ; Protein Structure, Tertiary ; SARS Virus ; genetics ; immunology ; Sequence Analysis, DNA ; Sequence Homology ; Spike Glycoprotein, Coronavirus ; Viral Envelope Proteins ; genetics ; metabolism

Lycophytes and seed plants constitute the typical vascular plants. Lycophytes have been thought to have no paleo-polyploidization although the event is known to be critical for the fast expansion of seed plants. Here, genomic analyses including the homologous gene dot plot analysis detected multiple paleo-polyploidization events, with one occurring approximately 13-15 million years ago (MYA) and another about 125-142 MYA, during the evolution of the genome of Selaginella moellendorffii, a model lycophyte. In addition, comparative analysis of reconstructed ancestral genomes of lycophytes and angiosperms suggested that lycophytes were affected by more paleo-polyploidization events than seed plants. Results from the present genomic analyses indicate that paleo-polyploidization has contributed to the successful establishment of both lineages-lycophytes and seed plants-of vascular plants.
Evolution, Molecular ; Genome, Plant ; Genomics ; Phylogeny ; Polyploidy ; Selaginellaceae/genetics*