OBJECTIVETo establish and optimize the two-dimensional electrophoresis (2-DE) of uterine leiomyoma for the proteome analysis.

METHODSRun immobilized pH gradient (IPG)-isoelectric focusing electrophoresis as the first dimension, then vertical SDS-PAGE electrophoresis as the second dimension. A series of important steps,such as sample solubility, volume of loading, electrophoresis parameters and protocol for staining were optimized.

RESULTSThe 2-DE patterns of uterine leiomyoma and myometrium with good quality were obtained.

CONCLUSIONWith optimal condition the two-dimensional electrophoresis of uterine leiomyoma can be obtained.

Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Leiomyoma ; chemistry ; Myometrium ; chemistry ; Neoplasm Proteins ; analysis ; Proteome ; analysis ; Uterine Neoplasms ; chemistry

OBJECTIVETo study the anti-cataract effect of gigantol combined with syringic acid and their action mechanism.

METHODH202-induced lens oxidative injury in vitro rat model was establish to observe the impact of gigantol combined with syringic acid on lens transparency under a dissecting microscope. D-galactose-induced cataract rat model was established to observe the impact of gigantol combined with syringic acid on lens transparency under a slit-lamp. UV spectrophotometry was adopted to detect the inhibitory activity of gigantol combined with syringic acid against AR. Molecular docking method was used to detect binding sites, binding types and pharmacophores of gigantol combined with syringic acid in prohibiting aldose reductase.

RESULTBoth in vitro and in vivo experiments showed a good anti-sugar cataract activity in the combination of gigantol and syringic acid and a better collaborative effect than single component-gigantol and syringic acid and positive control drug Catalin. Molecular docking and dynamic simulation showed their collaborative AR-inhibiting amino acid residue was Asn160 and the major acting force was Van der Waals' force, which formed common pharmacophores.

CONCLUSIONGigantol combined with syringic acid shows good anti-cataract, their action mechanism is reflected in their good collaborative inhibitory effect on AR.

Aldehyde Reductase ; antagonists & inhibitors ; Animals ; Bibenzyls ; Cataract ; drug therapy ; enzymology ; Drug Synergism ; Female ; Gallic Acid ; analogs & derivatives ; pharmacology ; Guaiacol ; analogs & derivatives ; pharmacology ; Humans ; In Vitro Techniques ; Lens, Crystalline ; drug effects ; enzymology ; Male ; Rats ; Rats, Wistar

OBJECTIVETo construct the delta-pIRES2-EGFP plasmid and investigate its expression in HEK293 cells.

METHODSFull length cDNA of rat delta opioid receptor gene amplified from rat brain tissues using reverse transcription and nested PCR was cloned into pMD20 T vector. The delta cDNA was inserted into pIRES2-EGFP plasmid to construct the recombinant eukaryotic plasmid delta-pIRES2-EGFP, which was transfected into HEK293 cells via Lipofectamine2000. The expression of delta was examined under fluorescence microscope.

RESULTSThe recombinant delta-pIRES2-EGFP plasmid was successfully constructed, and high expression of delta was detected in HEK293 cells transfected by the plasmid.

CONCLUSIONdelta-pIRES2-EGFP has been successfully cloned, which shows high expression of delta in HEK293 cells.

Animals ; DNA, Complementary ; genetics ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Polymerase Chain Reaction ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, delta ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Leaf senescence is often caused by water deficit and the chimeric gene P(SAG12)-IPT is an auto-regulated gene delaying leaf senescence. Using in vitro leaf discs culture system, the changes of contents of chlorophylls, carotenoids, soluble protein and thiobarbituric acid reactive substance (TBARS) and antioxidant enzymes activities were investigated during leaf senescence of P(SAGl2)-IPT modified gerbera induced by osmotic stress compared with the control plant (wild type). Leaf discs were incubated in 20%, 40% (w/v) polyethylene glycol (PEG) 6000 nutrient solution for 20 h under continuous light [130 micromol/(m(2) x s)]. The results showed that the contents of chlorophylls, carotenoids and soluble protein were decreased by osmotic stress with the decrease being more pronounced at 40% PEG, but that, at the same PEG concentration the decrease in the transgenic plants was significantly lower than that in the control plant. The activities of superoxide dismutase (SOD), catalases (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and dehydroascorbate reductase (DHAR) were stimulated by PEG treatment. However, the increases were higher in P(SAG12)-IPT transgenic plants than in the control plants, particularly at 40% PEG treatment. Lipid peroxidation (TBARS content) was increased by PEG treatment with the increase being much lower in transgenic plant than in the control plant. It could be concluded that the increases in the activities of antioxidant enzymes including SOD, CAT, APX, GPX and DHAR were responsible for the delay of leaf senescence induced by osmotic stress.
Alkyl and Aryl Transferases ; genetics ; Antioxidants ; metabolism ; Arabidopsis Proteins ; genetics ; Ascorbate Peroxidases ; Asteraceae ; genetics ; metabolism ; Carotenoids ; metabolism ; Catalase ; metabolism ; Chlorophyll ; metabolism ; Cysteine Endopeptidases ; genetics ; Genes, Bacterial ; Genes, Plant ; Lipid Peroxidation ; Osmotic Pressure ; Oxidoreductases ; metabolism ; Peroxidase ; metabolism ; Peroxidases ; metabolism ; Plant Leaves ; metabolism ; Plant Proteins ; metabolism ; Plants, Genetically Modified ; Promoter Regions, Genetic ; Solubility ; Superoxide Dismutase ; metabolism

Objective To evaluate the clinical outcome of periapical endodontic surgery for teeth that can't be treated by nonsurgical endodontic methods. Methods Sixty-two affected teeth were chosen for surgical endodontic treatment, of which 31 teeth underwent periapical curettage and the others were treated by root-end resection, retrograde preparation and filling. A radiography was taken immediately after surgery and was compared with those taken at 12 and 24 months. The results of two groups were analyzed using the X2 test. Results The success rate for retrograde filling was higher (85% after 12 months, 88% after 24 months) compared with that of periapical curettage (52% after 12 months, 45% after 24 months). The difference in success rate between the two groups was statistically significant. Conclusions Ultrasonic root-end preparation and retrograde filling is a good choice of treatment when the teeth can't be treated appropriately by nonsurgical treatment.

OBJECTIVETo investigate the antimicrobial resistance of clinical isolates of Stenotrophomonas matophilia (SMA) and the mechanisms of their drug resistance.

METHODSDisc diffusion method (NCCLS) was used to detect the resistant patterns of 88 initial SMA isolates resistant to 12 antibiotics isolated from a local hospital in the past 4 years. PCR was used to detect the 7 aminoglycosides modifying enzymes genes (AME) against amikacin and gentamicin. Metal-beta-lactamases (MBLs) were screened by synergic method, and extended-spectrum beta-lactamases (ESBLs) were detected by double-disk synergy test.

RESULTSThe resistance rates of the SMA isolates were 0%-9.7% to minocycline, 12.5%-22.6% to ticarcillin-clavulanic acid, 12.5%-28.6% to levofloxacin, 18.8%-33.3% to doxycycline, 18.8%-40% to sulfamethoxazole compound, 50%-65.7% to ciprofloxacin, 50%-66.7% to cehazindme, 54.8%-66.7% to amikacin, 75%-100% to gentamicin, 81.3%-100% to piperacillin, 87.5%-100% to aztreonam and 93.5%-100% to imipenem. Aac(3)-I and ant(4')-II were not detected in these strains. The positive rates of the other 5 AME genes of aac(3)-II, ant(2'')-I, aac(6')-I, aac(3)-III, aac(3)-IV were 2.3%, 5.7%, 8%, 10%, and 10%, respectively. SMA strains producing ESBLs were found at the rate of 38.6%; 25% of the strains were MBL-producing, and 13.6% produced both ESBLs and MBLs.

CONCLUSIONMost of the SMAs we isolated are multidrug-resistant through various mechanisms. The choice of antibiotics should be made according to the susceptibility results.

Amikacin ; pharmacology ; Drug Resistance, Multiple, Bacterial ; Gentamicins ; pharmacology ; Humans ; Imipenem ; pharmacology ; Microbial Sensitivity Tests ; Stenotrophomonas maltophilia ; drug effects ; isolation & purification

To investigate the effect of hepatitis B virus X protein(HBx) on CtBP-interacting protein(CtIP) which is an important repair factor of DNA double strand break damage in HepG2 cells induced by bleomycin. A HBx stably expressing HepG2 cell line and a control HepG2 cell line with empty vector transfected were established. After the double strand break (DSB) damage occurred, the mRNA and protein levels of CtIP were detected by Real-time PCR and Western blot assay respectively, cell cycle profiles and apoptotic cell death were determined by a flow cytometry, and the position of CtIP in cells was observed by confocal laser scanning microscopy. It showed that HepG2 cells transfected with hepatitis B virus X gene could stably express HBx protein. After being induced by bleomycin, the percentage of apoptotic cell was 16.90%+/-0.89% in HBx stably expressing HepG2 cell line and 15.30%+/-0.86% in control cell line, respectively (q = 2.074, P is more than to 0.05). While the percentage of death cell was 8.71%+/-0.74% in HBx stably expressing HepG2 cell line and 4.90%+/-0.46% in control cell line, respectively (q = 7.126, P is less than to 0.01). The two cell lines manifested the increase of G2/M arrest and significant difference existed between the two cell lines. HBx down regulated the expression levels of CtIP and its mRNA. The CtIP level was 0.66+/-0.04 in HepG2-HBx cell and 0.73+/-0.05 in HepG2-vec cell, respectively (t = 2.314, P is less than to 0.05). The relative mRNA level was 1.00+/-0.06 in HepG2-HBx cell and 1.23+/-0.08 in HepG2-vec cell, respectively (t = 2. 732, P is less than to 0.05). We also found that CtIP was concentrated in the cell nucleus. The research suggests that HBx may affect DNA-repair pathways by disrupting the expression of CtIP.
Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; metabolism ; Real-Time Polymerase Chain Reaction

To study and analyze prevalence condition of hypertension and complicated cardiovascular risk factors in north Fujian rural area .Methods : By cluster sampling , an epidemiological questionnaire and physical ex‐amination were performed among 1784 residents using standardized survey method , who were ≥35 years from six towns and eight villages in north Fujian rural area from Jun 2013 to Mar 2014. Hypertension was diagnosed accord‐ing to The 2010 Chinese Guideline for the Prevention and Treatment of Hypertension , multivariate Logistic regres‐sion was used to analyze independent risk factors for hypertension in these subjects .Results : Prevalence rate of hy‐ pertension was 17.43% (31／1784 ) in north Fujian rural area .After population standardization , it＇s 23. 21%(18.66% in men and 16. 13% in women) , there was no significant difference in prevalence rate of hypertension be‐tween men and women in any age layer , P＞0. 05 all.Multivariate Logistic regression analysis indicated that age , overweight (BMI≥24 kg／m2 ) , high triglyceride and fasting blood glucose were independent risk factors for hyper‐tension in north Fujian rural area (OR＝1.107－2.096 , P＜0. 05 or ＜0. 01).Conclusion : Prevalence rate of hyper‐tension is high in north Fujian rural area , age , overweight , smoking , high triglyceride and fasting blood glucose are the main risk factors .

Influenza virus is the causative agent of the seasonal and occasional pandemic flu. The current H1N1 influenza pandemic, announced by the WHO in June 2009, is highly contagious and responsible for global economic losses and fatalities. Although the H1N1 gene segments have three origins in terms of host species, the virus has been named swine-origin influenza virus (S-OIV) due to a predominant swine origin. 2009 S-OIV has been shown to highly resemble the 1918 pandemic virus in many aspects. Hemagglutinin is responsible for the host range and receptor binding of the virus and is therefore a primary indicator for the potential of infection. Primary sequence analysis of the 2009 S-OIV hemagglutinin (HA) reveals its closest relationship to that of the 1918 pandemic influenza virus, however, analysis at the structural level is necessary to critically assess the functional significance. In this report, we report the crystal structure of soluble hemagglutinin H1 (09H1) at 2.9 Å, illustrating that the 09H1 is very similar to the 1918 pandemic HA (18H1) in overall structure and the structural modules, including the five defined antiboby (Ab)-binding epitopes. Our results provide an explanation as to why sera from the survivors of the 1918 pandemics can neutralize the 2009 S-OIV, and people born around the 1918 are resistant to the current pandemic, yet younger generations are more susceptible to the 2009 pandemic.
Animals ; Cloning, Molecular ; Crystallography, X-Ray ; Hemagglutinin Glycoproteins, Influenza Virus ; chemistry ; genetics ; immunology ; isolation & purification ; Influenza A Virus, H1N1 Subtype ; chemistry ; genetics ; immunology ; Models, Molecular ; Protein Conformation ; Swine ; virology

Objective To explore and analyze the expression and prognostic value of serum vitamin D(VitD),fer-ritin(FRT)and heparin-binding epidermal growth factor(HB-EGF)in sepsis patients.Methods 86 sepsis patients who were admitted to the intensive care unit(ICU)of a hospital from January 2021 to January 2022 were selected as the case group,and 60 non-sepsis patients in the ICU were selected as the control group.According to the prognosis of sepsis patients one month later,patients were divided into survival group and death group.Patient's serum upon admission was taken,levels of serum VitD,FRT and HB-EGF were detected,the correlation with the prognosis of sepsis patients was analyzed,and the prognostic value was evaluated by area under curve(AUC)of receiver opera-ting characteristic curve.Results Levels of white blood cell count(WBC),C-reactive protein(CRP),procalcitonin(PCT),tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-1β and FRT of case group were all higher than those of non-sepsis patients in the control group,while the VitD and HB-EGF levels were lower than those of non-sepsis patients in the control group,differences were all statistically significant(all P＜0.05).One month follow-up on the prognosis of sepsis patients showed that 55 patients survived and 31 died.Acute physiology and chronic health evaluationⅡ(APACHE IⅡ)score,sequential organ failure assessment(SOFA)score,PCT,TNF-α,L-1βand FRT score in patients in the death group were all higher than those in patients in the survival group,while VitD and HB-EGF were lower than patients in the survival group,differences were all statistically significant(all P＜0.05).The Pearson correlation analysis results showed that VitD were negatively correlated with APACHE Ⅱ score,SOFA score,WBC,CRP,PCT and TNF-α(all P＜0.05),HB-EGF was negatively correlected with APACHE Ⅱ,score,CRP,PCT,TNF-α,IL-6 and IL-1β(all P＜0.05);while FRT was positively correlated with APACHE Ⅱscores,CRP,PCT,TNF-α,IL-6 and IL-1β(all P＜0.05).The AUC,sensitivity,and specificity of combined de-tection of serum VitD,FRT and HB-EGF in predicting the prognosis of sepsis patients were 0.82(95％CI:0.72-0.86),84.39％,and 69.35％,respectively.Conclusion Serum levels of VitD and HB-EGF are lower and FRT is higher in sepsis patients,their expression levels are closely related to patient prognosis,and have good predictive value for predicting the prognosis of sepsis patients.