Objective: To study the inhibitory effect of retrovirus-mediated antisense human telomerase RNA (hTR) gene on hepatocelluar carcinoma, so as to explore an effective way to inhibit telomrerase activity in the treatment of hepatocellular carcinoma. Methods: Sense and antisense hTR gene were transfected into the packaging cell line PT67 by electroporation, and the stablely transfected cells were selected with G418. The recombinant retroviral supernatant was collected and transfected into HepG2 cells. After G418 selection, PCR was used to verify the integration of the hTR gene. Cell growth curves were drawn using MTT assay and the expression of PCNA was determined by immunofluorescence. TRAP-PCR-ELISA was adopted to detect the telomerase activity; cell cycle and apoptosis were evaluated by flow cytometry (FCM).Results: The expression of hTR gene could be amplified in HepG2-hTR-EcoRⅠ and HepG2-hTR-BamHⅠ cells, but not in untransfected HepG2 cells. The antisense hTR complementary to the template region of telomerase inhibited growth and proliferation of HepG2 cells. The expression of neutrophil proliferating cell nuclear antigen (PCNA) was decreased. Telomerase activity in the antisense hTR-treated group was (2.31?0.16),which was significantly lower than those of the other 2 groups(P

Objective:To compare the effect of intracorporeal anastamosis and extracorporeal anastamosis on abdominal infection associated with laparoscopic right hemicolectomy.Methods:The clinical data of 210 patients with colon cancer who underwent laparoscopic right hemicolectomy in Dalian Third Peoples′s Hospital, Liaoning Province from January 2015 to December 2019 were analyzed retrospectively.Among them, 79 patients underwent intracorporeal anastamosis (intracorporeal anastamosis group) and 131 patients underwent extracorporeal anastamosis (extracorporeal anastamosis group). The perioperative indexes and postoperative abdominal infection were compared between the two groups.Results:In intracorporeal anastamosis group, the intraoperative bleeding was (45.2±4.2) mL, the operative time was (161.3±22.4) min, the number of lymph node dissection was (30.8±9.6), the postoperative exhaust time was (3.3±1.2) d, and the postoperative hospital stay was (7.6±0.5) d. In extracorporeal anastamosis group was (42.1±5.0) mL, (167.3±26.7) min, (32.9±8.6), (3.4±1.0) d and (7.5±0.6) d, respectively, there were no significant difference between the two groups (t value were 0.417, 0.207, 0.829, 0.338 and 0.293, respectively; P value were 0.699, 0.845, 0.231, 0.734 and 0.802, respectively). In intracorporeal anastamosis group, the incidence of abdominal infection (with anastomotic fistula)was 13.9%(11/79), the incidence of abdominal infection (without anastomotic fistula)was 10.1%(8/79), and in extracorporeal anastamosis group was 1.5%(2/131)and 0.8%(1/131), the differences were statistically significant (χ 2=12.805, 10.238; P=0.003, 0.008). In intracorporeal anastamosis group, the incidence of respiratory system infection was 1.3%(1/79), the incidence of urinary system infection was 2.5%(2/79), the incidence of surgical incision infection was 1.3%(1/79). In extracorporeal anastamosis group was 3.1%(4/131), 0.8%(1/131) and 3.1%(4/131), respectively.There were no significant difference between the two groups (χ 2 value were 0.662, 0.420 and 0.662, respectively; P value were 0.364, 0.587 and 0.364, respectively). Conclusion:Laparoscopic right hemicolectomy with intracorporeal anastamosis and extracorporeal anastamosis have the same surgical effect, but intracorporeal anastamosis may increase the risk of postoperative abdominal infection.

Objective:To explore the effects of MiR-199a-3p on promoting the development of mycoplasma pneumonia via the silent information regulator 1(SIRT1)/nuclear factor-κB (NF-κB) pathway.Methods:Totally 80 SPF-grade BALB/c mice were randomly divided into a control group, a model group, a miR-199a-3p group, and a inhibitory group, with 20 rats in each group. Excep the control group, the model group was established as a mouse model of mycoplasma pneumoniae through a continuous nasal drip of a high-load mycoplasma pneumoniae bacterial solution for 3 days. After modeling, mice in each group had tail vein injections. The miR-199a-3p group and the inhibitory group mice were injected with agomir solution and antagonir solution, respectively. And the model group and control group mice were injected with the same amount of physiological saline through the tail vein. After the experiment, mice of all groups were killed and their blood was collected from the eyeballs, and interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) levels were detected by the enzyme-linked immunosorbent assay method. Subsequently, the lung tissues of the mice were taken for HE staining to observe pathological changes in the lung tissue. Real-time fluorescence quantitative PCR was used to detect miR-199a-3p gene expression levels in lung tissue, and Western Blot was used to detect SIRT1/NF-κB signaling pathway protein expression in lung tissue. Results:Compared with the model group, IL-4, IL-6, IL-1β, and TNF-α levels in the serum of the miR-199a-3p group were decreased, with a significant difference ( P < 0.05). The HE staining results showed that the lung tissue structure of the control group mice was nearly normal and there was no alveolar exudation or inflammatory response. The other three groups all had varying degrees of alveolar interstitial widening, alveolar exudation, and inflammatory cell infiltration. Compared with the model group, miR-199a-3p gene and SIRT1 protein expression levels in the miR-199a-3p group increased, with a significant difference (all P < 0.05). While NF-κB protein expression levels in the miR-199a-3p group decreased, there was a significant difference ( P < 0.05). Conclusions:The expression of miR-199a-3p gene is reduced in the lung tissue of mycoplasma pneumoniae mice. Increasing the level of miR-199a-3p gene has a protective effect on lung tissue damage in mycoplasma pneumoniae mice through anti-inflammatory effects, and its mechanism may be related to its regulatory effect on the SIRT1/NF-κB pathway.

Coloring Agents ; Diagnostic Uses of Chemicals ; Esophageal Neoplasms ; diagnosis ; Esophagoscopy ; Esophagus ; pathology ; Humans ; Iodine

Ischemia-reperfusion injury is one of the main causes of complications related to liver transplantation, hepatectomy, trauma and hemorrhagic shock. The cells of ischemia and hypoxic injury release of injury-related molecular patterns, lead to the activation of immune cells and cytokine, which further aggravates the inflammatory response and enlarges the injury. It’s indicated that injury-related molecular patterns, liver resident immune cells and cytokines play a key role in promoting inflammation and liver ischemia-reperfusion injury. However, recent studies suggested that the ischemia cells and cytokines played acomplex role in this process. Relevant progresses were reviewed in this article.

Objective To evaluate the ability of Matrix-assisted laser desorption ionization-time of flight mass spectrum (MALDI-TOF MS) on homology analysis of carbapenem-resistant Klebsiella pneumoniae(Carbapenemase-Resistant K. pneumoniae, CRKp). Methods During January 2017 to December 2017, a total of 57 strains isolatedfromChanghai Hospital, Shanghai and 36 strainsfrom Urumqi First Hospital, Xinjiang were included for retrospective study. Inclusion criteria:strains which resistant to one of the carbapenems and with more than 100 peaks in identification mass spectrum. A total of 39 isolates were selected and divided into three groups according to the PFGE typing results:(1) Highly homologous group (17 strains with blaOXA-48 gene); (2) Moderate homology group (16 strains with blaKPC gene); (3)The non-homology group (8 strains with other resistance mechanisms). Mass spectrometry results were analyzed using BioNumeric and the two classification methods in SARAMIS, respectively.The PFGE analysis results were regarded as standard when evaluate the reliability of the other three analytical methods. Results Forhighly homologous group, the analysisresults of KA-A type dientical mass by two classification methods in SARAMIS were accord with PFGE,and the identity mass value is>80％, the similarity is>85％. However, for non-homologous group, the identity mass value is<80％, the similarity is<85％ when compared with PFGE, which indicated that the method based on MALDI-TOF MS was not premium for this group of strains. For moderate homologous strains,the results of two SARAMIS methods for KC-A type havepoor consistency with PFGE, for the identity mass value is<70％, the similarity is<90％. Conclusions Based on MALDI-TOF MS using BioNumeric software and SARAMIS database software could be regarded as a screening method for phylogenetic analysis among CRKp strains with high homology. While, it should not be applied to evaluate homology of the strains with genetic diversity for lacking stability.

PURPOSE: Cathepsin K is a potent collagenase implicated in human and animal atherosclerosis-based vascular remodeling. This study examined the hypothesis that serum CatK is associated with the prevalence of coronary artery disease (CAD). MATERIALS AND METHODS: Between January 2011 and December 2012, 256 consecutive subjects were enrolled from among patients who underwent coronary angiography and percutaneous coronary intervention treatment. A total of 129 age-matched subjects served as controls. RESULTS: The subjects' serum cathepsin K and high sensitive C-reactive protein (hs-CRP) and high-density lipoprotein cholesterol were measured. The patients with CAD had significantly higher serum cathepsin K levels compared to the controls (130.8+/-25.5 ng/mL vs. 86.9+/-25.5 ng/mL, p<0.001), and the patients with acute coronary syndrome had significantly higher serum cathepsin K levels compared to those with stable angina pectoris (137.1+/-26.9 ng/mL vs. 102.6+/-12.9 ng/mL, p<0.001). A linear regression analysis showed that overall, the cathepsin K levels were inversely correlated with the high-density lipoprotein levels (r=-0.29, p<0.01) and positively with hs-CRP levels (r=0.32, p<0.01). Multiple logistic regression analyses shows that cathepsin K levels were independent predictors of CAD (odds ratio, 1.76; 95% confidence interval, 1.12 to 1.56; p<0.01). CONCLUSION: These data indicated that elevated levels of cathepsin K are closely associated with the presence of CAD and that circulating cathepsin K serves a useful biomarker for CAD.
Aged ; Biological Markers/blood ; C-Reactive Protein/metabolism ; Cathepsin K/*blood ; Coronary Artery Disease/*blood/metabolism ; Female ; Humans ; Male ; Middle Aged

PURPOSE: Cathepsin K is a potent collagenase implicated in human and animal atherosclerosis-based vascular remodeling. This study examined the hypothesis that serum CatK is associated with the prevalence of coronary artery disease (CAD). MATERIALS AND METHODS: Between January 2011 and December 2012, 256 consecutive subjects were enrolled from among patients who underwent coronary angiography and percutaneous coronary intervention treatment. A total of 129 age-matched subjects served as controls. RESULTS: The subjects' serum cathepsin K and high sensitive C-reactive protein (hs-CRP) and high-density lipoprotein cholesterol were measured. The patients with CAD had significantly higher serum cathepsin K levels compared to the controls (130.8+/-25.5 ng/mL vs. 86.9+/-25.5 ng/mL, p<0.001), and the patients with acute coronary syndrome had significantly higher serum cathepsin K levels compared to those with stable angina pectoris (137.1+/-26.9 ng/mL vs. 102.6+/-12.9 ng/mL, p<0.001). A linear regression analysis showed that overall, the cathepsin K levels were inversely correlated with the high-density lipoprotein levels (r=-0.29, p<0.01) and positively with hs-CRP levels (r=0.32, p<0.01). Multiple logistic regression analyses shows that cathepsin K levels were independent predictors of CAD (odds ratio, 1.76; 95% confidence interval, 1.12 to 1.56; p<0.01). CONCLUSION: These data indicated that elevated levels of cathepsin K are closely associated with the presence of CAD and that circulating cathepsin K serves a useful biomarker for CAD.
Aged ; Biological Markers/blood ; C-Reactive Protein/metabolism ; Cathepsin K/*blood ; Coronary Artery Disease/*blood/metabolism ; Female ; Humans ; Male ; Middle Aged

OBJECTIVETo select and identify ssDNA aptamers specific to Streptococcus mutans strains with different cariogenicity isolated from clinical specimens.

METHODSSubtractive SELEX technology targeting the whole intact cells was used to screen for ssDNA aptamers specific to the clinical isolates Streptococcus mutans strains with different cariogenicity. Radioactive isotope, flow cytometry, gene cloning and sequencing, MEME online software and RNA structure analysis software were employed to analyze the first and secondary structures of the aptamers and identify the screened aptamers.

RESULTSDetection by radioactive isotope showed sufficient pool enrichment after 9 rounds of subtractive SELEX. Flow cytometry showed that the selected aptamers H1, H16, H4, L1, L10 and H19 were capable of binding specifically with highly cariogenic Streptococcus mutans strains but not with strains with a low cariogenicity. The aptamer H19 had the strongest binding capacity to highly cariogenic Streptococcus mutans strains, with a dissociation constant of 69.45∓38.53 nmol/L.

CONCLUSIONWe have obtained the ssDNA aptamers specific to the clinical isolates of highly cariogenic Streptococcus mutans strains.

Aptamers, Nucleotide ; genetics ; Cloning, Molecular ; DNA Primers ; Dental Caries ; microbiology ; Gene Library ; Humans ; Nucleic Acid Conformation ; SELEX Aptamer Technique ; Species Specificity ; Streptococcus mutans ; classification ; genetics ; isolation & purification

OBJECTIVETo screen of high cariogenicity Streptococcus mutans (S. mutans) strains isolated from clinical specimens preliminary.

METHODSAcidogenicity, aciduricity, extracellular polysaccharide production and adhesion of 41 strains of S. mutans isolated from clinical specimens were investigated to screen high cariogenicity S. mutans strains.

RESULTSThere were different cariogenicity among 41 strains of S. mutans, in which 3 strains of S. mutans had all high ability to produce extracellular polysaccharide, adhere to the saliva-coated hydroxyapatite, produce acid and tolerate acid, indicated there were 3 strains with high cariogenicity S. mutans strains isolated from clinical specimens. Another 3 strains of S. mutans with all low ability to produce extracellular polysaccharide, adhere to the saliva-coated hydroxyapatite, produce acid and tolerate acid indicated they were low cariogenicity S. mutans strains isolated from clinical specimens.

CONCLUSIONWe may have obtained high cariogenicity S. mutans strains isolated from clinical specimens.

Dental Caries ; Durapatite ; Humans ; Saliva ; Streptococcus mutans