Objective To investigate the effects of SIRT1 gene expression inhibited by shRNA on proliferation and apoptosis of pancreatic carcinoma PANC1 cells. Methods The eukaryotic expression plasmid of short hairpin RNA (shRNA) targeting SIRT1 gene (SIRT1-shRNA) was constructed as pGC-shRNA and transfected into PANC1 cells. There were shRNA-control transfection group and untransfectian control group. The expressions of SIRT1 mRNA and protein were detected with real-time PCR and immunocytochemistry assay, respectively. The growth rate of PANC1 cells was detected by MTT. Activity of caspase-3 and caspase-9 were detected by ELASA. Expressions of Bax, Bcl-2 proteins were determined by Western blotting. ResultsCompared with untransfected group, the inhibition rate of expressions of SIRT1 mRNA and protein were (76.2 ± 10.4) % and (80.1 ± 11.6) %, cytostasis rate was (45.1 ± 6.5) %, caspase-3 and caspase-9 activity was significantly increased, Bax protein was up-regulated, while Bcl-2 protein was down-regulated 48h after transfection. Conclusions Recombinant expression plasmid SIRT1 shRNA could significantly inhibit the expression of SIRT1 gene, and the mechanism may include increased caspase-3 and caspase-9 activity and Bax protein up-regulation, as well as Bcl-2 down-regulation.

Objective To investigate the effect of resveratrol on the proliferation and invasion of human pancreatic cancer PANC-1 cells. Methods Five groups including blank control group, 0. 1% dimethylsulfoxide (DMSO) group and resveratrol groups (50, 100, 200 μmol/L) were established. The proliferation of PANC-1 cells was detected by MTT assay. The apoptosis and cell cycle change were analyzed by flow cytometry. The invasive ability of PANC-1 cells was observed with a Transwell cell culture chamber. The expressions of Bax, Bcl-2,matrix metalloproteinases 2 (MMP-2) and 9 (MMP-9) of the PANC-1 cells were assayed by real-time quantitative PCR and Western blot. All data were analyzed using the analysis of variance. Results ( 1 ) The inhibition rate of resveratrol on the proliferation of PANC-1 cells was 0 in the blank control group, 3.25% ±0.42% in the 0. 1% DMSO group, 13.23% ± 1.68% in the 50 μmol/L of resveratrol group, 42.25% ± 3.20% in the 100 μmol/L of resveratrol group, and 56.94% ±5.31% in the 200 μmol/L of resveratrol group. There was a significant difference in the inhibition rate among the five groups (F=460. 10, P<0.05). (2) The apoptosis rate was 0.05% ±0.03% in the blank control group, 3.39% ± 1.77% in the 0. 1% DMSO group, 6.92% ± 1.85% in the 50 μmol/L of resveratrol group, 19.05% ± 2.01% in the 100 μmol/L of resveratrol group, and 27. 17% ±6.43% in the 200 μmol/L of resveratrol group. There was a significant difference in the apoptosis rate among the five groups (F = 38.84, P < 0.05). (3) There was no significant effect of 0. 1% DMSO on the cell cycle of PANC-1 cells. The number of PANC-1 cells in the G0/G1 and S phase was increased. (4) The average number of invading PANC-1 cells was 61 ± 13 in the blank control group, 54 ± 13 in the 0. 1% DMSO group, 48 ± 15 in the 50 μmol/L of resveratrol group, 23 ±6 in the 100 μ mol/L of resveratrol group and 18 ±7 in the 200 μmol/L of resveratrol group. There was a significant difference in the number of invading PANC-1 cells among the five groups (F = 69.08, P < 0.05 ). (5) There were up-regulated mRNA and protein expressions of Bax and down-regulated mRNA and protein expressions of Bcl-2, and the expressions of MMP-2 and MMP-9 of the PANC-1 cells were inhibited in the resveratrol groups. The changes of the protein expressions of Bax, Bcl-2, MMP-2, MMP-9 were consistent with the changes of the mRNA expressions of the four indexes. Conclusion Resveratrol can significantly inhibit the proliferation and invasion, as well as induce apoptosis of PANC-1 cells in vitro.

BACKGROUND:C-reactive protein has been shown to rapidly increase during the occurrence of inflammation and tissue injury, and can indicate the degree of inflammatory reaction.
OBJECTIVE:To analyze the correlation between survival time and C-reactive protein in rabbits after transplantation of polytetrafluoroethene artificial trachea with support ring.
METHODS:The cervical trachea of rabbits was replaced by polytetrafluoroethene artificial trachea with support ring. Survival time of the rabbit, and the changes in serum C-reactive protein at 1-7 days after transplantation were observed. Linear regression was used to assess the univariate association between serum C-reactive protein and survival time.
RESULTS AND CONCLUSION:The linear correlation was observed between changes of serum C-reactive protein and survival time in rabbits with artificial trachea replacement operation. C-reactive protein levels in rabbits with<13 days of survival time were increased and positively associated with the number of days after transplantation. However, C-reactive protein levels in rabbits with>13 days were decreased and negatively associated with the number of days after transplantation. In rabbits with positive correlation and negative correlation, the median survival time and 95%confidence interval (CI) were respectively 10 days (95%CI 8.614-11.386 days) and 27 days (95%CI 23.970-30.030 days). The survival rate in negative correlation group was significantly higher than positive correlation group (x2=29.364, P<0.01). Results suggested that the prolonged survival time of rabbits after artificial trachea replacement operation was related to the decreased concentration of serum C-reactive protein.

OBJECTIVETo establish an appropriate animal model of uterine leiomyoma and to understand the pathogenesis of this disease.

METHODSMature female rats were intramuscularly injected with estradiol benzoate at 200 μg or 300 μg twice a week. After injection for 8 or 10 weeks, the rats were sacrificed. We measured the serum levels of estrogen (E(2)) and progesterone (P), evaluated ER and PR expression, and calculated the leiomyoma forming rate and mortality of the rats. Histological changes were compared between rat uterine leiomyoma and human uterine leiomyoma with HE staining. The optimal dose and duration of E(2) for induction of uterine leiomyoma in rat were determined.

RESULTSIn the rats treated with estradiol benzoate 200 μg for 8 weeks ìn the serum E(2) level increased significantly (P<0.01). Uterine nodules were visible in some of the tested rats. Based on the pathohistological Results , the uterine leiomyoma developed in the treated rats demonstrated similar features as in human uterine leiomyoma. The expressions of ER and PR were increased in the leiomyoma tissues.

CONCLUSIONThe rat model of uterine leiomyoma can be established by intramuscular injection of estradiol benzoate at 200 μg twice per week for 8 weeks, with similar features as those of human uterine leiomyoma. The high concentrations of ER and PR in uterine tissue might be related with the development of uterine leiomyoma in animal.

Animals ; Disease Models, Animal ; Estrogens ; administration & dosage ; adverse effects ; Female ; Leiomyoma ; chemically induced ; Rats ; Uterine Neoplasms ; chemically induced

Targeting rRNA of bacteria is a new strategy for antibiotic agent development. The rRNA such as mRNA are naturally self-folded molecules which expose only limited accessible target-sites for binding. These accessible sites are pivotal for designing the effective antisense oligonucleotides, ribozymes, and DNAzymes. MAST, an RNA accessible site screening method, illustrated 6 accessible sites on 16S rRNA by immobilizing 16S rRNA and hybridizing with oligonucleotide library. 5 of the accessible sites were identified valid, and the antisense oligonucleotides targeted to which showed inhibition effectiveness on the proliferation. Among the 5 target sites, one showed the priority of accessibility. Ribozyme designed to this site showed obvious inhibition to the growth when induced expressing in the transfection E.coli.

Abstract: Objective　To understand the distribution and drug resistance of bacteria in clinical blood culture specimens in Ningxia in recent years, and to provide a basis for the prevention and treatment of bloodstream infection diseases. 　　　Methods　The blood culture isolation bacteria and drug resistance of Ningxia bacterial resistance monitoring network hospitals from 2018 to 2020 were statistically analyzed by WHONET5.6 software. Results　In the past three years, a total of 6 757 strains of bacteria were isolated from blood samples, including 3 697 strains (54.7%) of gram-negative bacteria and 3 060 (45.3%) of gram-positive bacteria. Among the gram-negative bacteria, Escherichia coli (2 074 strains,30.7%), Klebsiella pneumoniae (696 strains), Pseudomonas aeruginosa (139 strains), and Acinetobacter baumannii (121 strains). Among the gram-positive bacteria, coagulase-negative Staphylococcus (1 691 strains,25.0%), Staphylococcus aureus (442 strains), Streptococcus spp. (431 strains), Enterococcus spp. (379 strains). Resistance to Escherichia coli and Klebsiella pneumoniae was 56.6% and 22.6% against third-generation cephalosporins, and resistance to carbapenems was 1.0% and 3.7%, respectively. Pseudomonas aeruginosa and Acinetobacter baumannii were resistant to carbapenems at 9.0%（12/139） and 80.7%（71/121）. Methicillin-resistant Staphylococcus aureus (MRSA) was detected at 26.8%, methicillin-resistant coagulase-negative Staphylococcus was detected at 70%, and no Staphylococcus bacteria resistant to vancomycin and linezolid were found. For three years, only 1 strain of vancomycin-resistant Enterococcus faecalis was detected, and no linezolid-resistant Staphylococcus and Enterococcus were detected. Conclusions　Ningxia clinical blood specimen isolates of Escherichia coli, coagulase-negative Staphylococcus, and Klebsiella pneumoniae are more common. Among them, the resistance rate of Escherichia coli and Klebsiella pneumoniae to the third generation of cephalosporins is relatively stable, and the resistance rate to carbapenems is low. Acinetobacter baumannii is highly resistant to carbapenems, and methicillin-resistant Staphylococcus aureus detection rates are on the rise and should be closely monitored.

OBJECTIVETo investigate the stability of curcumin, demethoxycurcumin and bisdemethoxycurcumin in different buffer solution.

METHODTo determine concentration of curcumin by HPLC when added curcumin, demethoxycurcumin and bisdemethoxycurcumin into the buffer solution the equation of degradation was established.

RESULTThe sequence of stability are bisdemethoxycurcumin > or = demethoxycurcumin > or =curcumin at the same condition.

CONCLUSIONThe demethoxycurcumin can stabilize curcumin more strong than the others. The demethoxycurcumin is a nature stabilizing agent for curcumin.

Chromatography, High Pressure Liquid ; Curcumin ; analogs & derivatives ; chemistry ; Drug Stability ; Hydrogen-Ion Concentration

Translocating protein and translocating peptides have therapeutic potential against tumors by exposing the cytotoxic domains of toxic proteins to the cell cytosol. The aim of this study is to investigate the effect of N-terminally fused PE translocating peptides on granzyme B (GrBa) activity. PE II-GrBa fusion protein genes were constructed by replacing N-terminal signal and acidic dipeptide sequence of human granzyme B gene with two truncated translocating sequences of Pseudomonas exotoxin A (PE II aa 280-364/358) by recombinant PCR, and then cloned into pIND inducible expression vector. The resulting pIND-PE II-GrBa expression vectors were co-transfected with assistant plasmid pVgRXR into HeLa cells through lipofectamine, followed by selection on G418 and zeocin. The resistant cells were collected and induced with ponasterone A. Western blot analysis demonstrated that ponasterone A induction caused the expression of PE II-GrBa fusion proteins, and indirect immunofluorescence detected giant sized multinucleated cells, suggesting cytoskeletal and mitotic abnormalities as reported in our previous studies. Western blot, enzymatic activity assay and cell counting analysis indicated that two types of PE II-GrBa fusion proteins were capable of cleaving both endogenous and exogenous substrates of granzyme B, and inhibiting the growth of cells. The PE II (aa 280-358)-GrBa was shown to have higher serine protease activity and stronger growth inhibitory effect. Such inhibition was presumably associated with G2 arrest as determined by cell cycle analysis. These data prove that PE II-GrBa fusion proteins have cell inhibitory effect similar to GrBa, and that the shorter PE-derived peptide exerts less influence on GrBa activity. This study helps to optimize the construction of recombinant protein comprising translocating peptides and cytotoxic molecules for tumor cell killing.
ADP Ribose Transferases ; genetics ; pharmacology ; Bacterial Toxins ; genetics ; pharmacology ; Cell Proliferation ; drug effects ; Exotoxins ; genetics ; pharmacology ; Granzymes ; genetics ; pharmacology ; HeLa Cells ; Humans ; Recombinant Fusion Proteins ; pharmacology ; Virulence Factors ; genetics ; pharmacology

OBJECTIVETo study the value of ultrasound elastography in the evaluation of ethanol-induced lesions of liver.

METHODSAlcohol with a dose of 2 ml was injected into a fresh porcine liver under ultrasound guidance to create stiff necrosis. Then freehand elastography of the lesion from the identical scan plane was obtained with Siemens SONOLINE Antares system using VF10-5 probe at about every 30 seconds till 6 minutes later. The original high-quality radio-frequency data were acquired through an ultrasound research interface provided by the ultrasound system. Corresponding elastograms were then produced offline using cross-corre-lation technique and compared with gross specimen.

RESULTSA hyperechoic area with acoustic shadow below appeared immediately after alcohol injection. The hyperechoic area diffused and its boundary was illegible following injection. On the contrary, the ethanol-induced lesion in elastography appeared as a low strain hard region surrounded by high-strain soft hepatic tissues with clear but irregular boundaries. Sequential elastograms with the lesion boundaries sketched showed that the lesion area grew in the first 3 minutes after ethanol injection and then reached a plateau, which corresponded to the gross specimen.

CONCLUSIONUltrasound elastography can be used to detect and evaluate the diffusion of ethanol-induced hepatic lesion.

Animals ; Disease Models, Animal ; Elasticity Imaging Techniques ; methods ; Ethanol ; adverse effects ; Humans ; Liver ; diagnostic imaging ; pathology ; Liver Diseases ; diagnosis ; diagnostic imaging ; pathology ; Swine

OBJECTIVETo evaluate the usefulness of contrast-enhanced ultrasonography (CEUS) in the diagnosis of biliary diseases.

METHODSCEUS was performed in 57 patients with biliary diseases. The contrast enhancement characteristics and the morphologic features were observed. The ultrasonographic results were compared with those obtained through conventional 2-D ultrasound (2D-US), color Doppler flow ultrasound (CDFI), and clinical, surgical, and laboratory findings. In some cases, the ultrasonographic results were also compared with those obtained from contrast-enhanced computed tomography (CECT).

RESULTSThe diagnostic accuracy of 2D-US combined with CEUS was significantly higher than that of 2D-US combined with CDFI 87.7% vs 71.9%; chi(2) = 4.41, P < 0.05). CEUS clearly showed the presence/absence of blood supply in biliary lesions and offered real-time imaging of the microcirculation perfusion in the lesions. It also offered useful information to differentiate biliary tumors from stones, bile mud, and/or blood clots. It distinctly displayed the size and contour of the lesions as well as the infiltrated range, depth, and the involved area. However, CEUS is most useful in reflecting blood perfusion patterns; it had limited value in differentiating the malignancies of polypoid lesions. The diagnostic accuracy (87.0% vs 91.3%;chi(2) = 0. 45, P > 0.05) and the size and range of the lesions displayed (0.4-6.2 cm vs 0.4-6.0 cm, P = 0.721) were not significantly different between CEUS and CECT.

CONCLUSIONCEUS is a useful tool in the routine ultrasonography of biliary diseases.

Adult ; Aged ; Aged, 80 and over ; Biliary Tract Diseases ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Sensitivity and Specificity ; Ultrasonography ; Young Adult