KAT8/MOF-Mediated Anti-Cancer Mechanism of Gemcitabine in Human Bladder Cancer Cells

Histone acetylation is a well-characterized epigenetic modification controlled by histone acetyltransferases (HATs) and histone deacetylases (HDACs). Imbalanced histone acetylation has been observed in many primary cancers. Therefore, efforts have been made to find drugs or small molecules such as HDAC inhibitors that can revert acetylation levels to normal in cancer cells. We observed dose-dependent reduction in the endogenous and exogenous protein expression levels of KAT8 (also known as human MOF), a member of the MYST family of HATs, and its corresponding histone acetylation at H4K5, H4K8, and H4K16 in chemotherapy drug gemcitabine (GEM)-exposed T24 bladder cancer (BLCA) cells. Interestingly, the reduction in MOF and histone H4 acetylation was inversely proportional to GEM-induced γH2AX, an indicator of chemotherapy drug effectiveness. Furthermore, pGL4-MOF-Luc reporter activities were significantly inhibited by GEM, thereby suggesting that GEM utilizes an MOF-mediated anti-BLCA mechanism of action. In the CCK-8, wound healing assays and Transwell ® experiments, the additive effects on cell proliferation and migration were observed in the presence of exogenous MOF and GEM. In addition, the promoted cell sensitivity to GEM by exogenous MOF in BLCA cells was confirmed using an Annexin V-FITC/PI assay. Taken together, our results provide the theoretical basis for elucidating the anti-BLCA mechanism of GEM.

Predictive factors for postoperative hypocalcemia after reoperation of thyroid cancer / 中华内分泌外科杂志

Objective Hypothyroidism(HypoPT) is one of the most common complications of thyroid reoperation,and hypocalcemia caused by HypoPT is a difficult problem in clinical practice.This study aims to investigate the risk factors for hypoealcemia after reoperation of thyroid cancer.Methods Data of 106 patients who underwent reoperation for thyroid cancer from 2013 to 2018 were retrospectively analyzed.According to the parathyroid area involved in the reoperation,the reoperation was graded:low-risk group,intermediate-risk group and high-risk group.Parathyroid hormone(PTH) and total calcium(Ca) levels were monitored at 12 to 24 hours after surgery.The follow-up period was at least 6 months.The PTH cut-off values of postoperative hypocalcemia were analyzed by ROC curve.The chi-square test and logistic regression analysis were used to analyze the risk factors of postoperative hypocalcemia.Results The incidence of transient HypoPT after reoperation was 35％,and the permanent HypoPT was 1.9％.The grade of reoperation was positively correlated with postoperative HypoPT.The ROC curve showed that the PTH cut-off values of postoperative hypocalcemia was 15 pg/ml and area under curve(AUC) was 0.636(95％ CI:0.530-0.742,P＜0.017).Univariate and multivariate analysis showed that reoperation grade and postoperative PTH＜15 pg/ml were independent risk factors for postoperative hypocalcemia.Conclusion The grade of reoperation and postoperative PTH value ＜15 pg/ml can predict postoperative hypocalcemia.

Over expression of AdeABC and AcrAB-TolC efflux systems confers tigecycline resistance in clinical isolates of Acinetobacter baumannii and Klebsiella pneumoniae

Abstract: INTRODUCTION: Due to the wide use of tigecycline in the treatment of severe infections caused by multidrug-resistant (MDR) bacteria, clinical resistance to tigecycline has increased in recent years. Here, we investigated the relationship between tigecycline resistance and the expression of efflux pumps. METHODS: Clinical isolates of Acinetobacter baumannii and Klebsiella pneumoniae were consecutively collected from hospitalized patients in three hospitals. The minimum inhibitory concentration (MIC) of tigecycline was determined using the broth microdilution method. Expression levels of efflux pump genes and regulators were examined by quantitative real-time reverse transcription polymerase chain reaction. The correlations between tigecycline MICs and gene expression levels were analyzed. RESULTS: Overall, 1,026 A. baumannii and 725 K. pneumoniae strains were collected. Most strains were isolated from sputum. The tigecycline resistance rate was 13.4% in A. baumannii isolates and 6.5% in K. pneumoniae isolates. Overexpression of AdeABC and AcrAB-TolC efflux systems was observed found in clinical tigecycline-resistant isolates. The tigecycline MIC had a linear relationship with the adeB expression level in A. baumannii isolates, but not with the acrB expression level in K. pneumoniae isolates. There were significant linear trends in the overexpression of ramA as the tigecycline MIC increased in K. pneumoniae isolates. CONCLUSIONS: Tigecycline resistance in A. baumannii and K. pneumoniae was strongly associated with the overexpression of efflux systems. More studies are needed to elucidate whether there are other regulators that affect the expression of adeB in A. baumannii and how ramA affects the expression of acrB in K. pneumoniae.

Human INO80/YY1 chromatin remodeling complex transcriptionally regulates the BRCA2- and CDKN1A-interacting protein (BCCIP) in cells

The BCCIP (BRCA2- and CDKN1A-interacting protein) is an important cofactor for BRCA2 in tumor suppression. Although the low expression of BCCIP is observed in multiple clinically diagnosed primary tumor tissues such as ovarian cancer, renal cell carcinoma and colorectal carcinoma, the mechanism of how BCCIP is regulated in cells is still unclear. The human INO80/YY1 chromatin remodeling complex composed of 15 subunits catalyzes ATP-dependent sliding of nucleosomes along DNA. Here, we first report that BCCIP is a novel target gene of the INO80/YY1 complex by presenting a series of experimental evidence. Gene expression studies combined with siRNA knockdown data locked candidate genes including BCCIP of the INO80/YY1 complex. Silencing or over-expressing the subunits of the INO80/YY1 complex regulates the expression level of BCCIP both in mRNA and proteins in cells. Also, the functions of INO80/YY1 complex in regulating the transactivation of BCCIP were confirmed by luciferase reporter assays. Chromatin immunoprecipitation (ChIP) experiments clarify the enrichment of INO80 and YY1 at +0.17 kb downstream of the BCCIP transcriptional start site. However, this enrichment is significantly inhibited by either knocking down INO80 or YY1, suggesting the existence of both INO80 and YY1 is required for recruiting the INO80/YY1 complex to BCCIP promoter region. Our findings strongly indicate that BCCIP is a potential target gene of the INO80/YY1 complex.

Nicotine alleviates the liver inflammation of non-alcoholic steatohepatitis induced by high-fat and high-fructose in mice / 北京大学学报(医学版)

Objective:To investigate the anti-inflammation effects by activation of the cholinergic anti-inflammatory pathway and its mechanisms in non-alcoholic steatohepatitis (NASH)model mice.Me-thods:6-week-old male C57BL/6J (B6)mice were randomly divided into four groups:the first group was normal mice,injected with saline;the second group was normal mice,injected with nicotine;the third group was NASH model mice,injected with saline;the fourth group was NASH model mice,injec-ted with nicotine.The experimental mice were fed with either standard chow (SC)or high-fat and high-fructose (HFHF)for 17 weeks to generate an NASH model mice.The mice received injection once daily for 3 weeks [nicotine dose,400 μg/kg].Then,their pathological characteristics and function of the liver were assessed.The expressions of interleukin-6 (IL-6)and tumor necrosis factor-α(TNF-α)in se-rum were analyzed by enzyme linked immunosorbent assay (ELISA).The expressions of alpha 7 nicotinic acetylcholine receptors (α7nAChR),Toll-like receptors-4 (TLR-4)and nuclear factor κB of phosphory-lation (p-NF-κB)in Kupffer cells were determined by Western blot and immunofluorescence assays.Re-sults:We successfully generated NASH model mice by imitating the high-fat and high-fructose dietary style of NASH patients.The results of our investigation demonstrated that nicotine could reduce signifi-cantly the levels of IL-6,and TNF-αin serum (P <0.05).The expression of p-NF-κB protein in the group which was NASH model mice injected with nicotine declined significantly as compared with the group which was NASH model mice injected with saline (P <0.05).And the expression of α7nAChR protein elevated significantly conversely (P <0.05 ).Conclusion:Activation of the cholinergic anti-inflammatory pathway could inhibit the release of inflammatory factors as TNF-αand IL-6 in NASH model mice,and the mechanism for the inhibition of inflammatory was mediated by NF-κB pathway.

Clinical research of chronic pelvic cavity pain syndrome treated with acupoint catgut embedding therapy / 中国针灸

<p><b>OBJECTIVE</b>To explore the clinical efficacy of acupoint catgut embedding therapy on chronic pelvic cavity pain syndrome differentiated as kidney deficiency and stagnation of damp heat, and explore the impacts on plasma P substance (SP), plasma beta-endorphin (β-EP).</p><p><b>METHODS</b>One hundred and eighty cases were randomly divided into a catgut embedding group (90 cases) and a western medication group (90 cases). In the western medication group, tamsulosin capsules 0. 2 mg were prescribed for oral administration, once a day; indometacin sustained release tablets, 25 mg, three times a day. Totally, the oral administration for 8 weeks was required. In the catgut embedding group, the acupoint catgut embedding therapy was applied to Qugu (CV 2), Shenshu (BL 23), Zhibian (BL 54), Huiyin (CV 1) and Sanyinjiao (SP 6), once every two weeks; the treatment of 4 weeks made one session, and two sessions were required. Before and after treatment, TCM symptom score, NIH-CPSI (the National Institute of Health Chronic Prostatitis Symptom Index) score, lecithin body numbers in prostatic fluid, score in SAS (self-rating anxiety scale), score in SDS (self-rating depression scale), the levels of SP and β-EP, etc. were observed in the two groups, and the clinical efficacy was assessed in the two groups.</p><p><b>RESULTS</b>(1) Ten cases were dropped in either group. The total effective rate was 91. 25% (73/80) in the catgut embedding group, higher than 78. 75% (63/80) in the western medication group (P<0. 05). (2) After treatment, TCM symptom score, total score in NIH-CPSI, pain score and the scores in SAS and SDS were all reduced as compared with those before treatment in the two groups (all P<0. 05). After treatment, TCM symptom score, total score and pain score in NIH-CPSI, and the scores in SAS and SDS in the catgut embedding group were both lower than those in the western medication group (all P<0. 05). (3) After treatment, the lecithin body numbers were both increased as compared with those before treatment in the two groups (both P<0. 05), and the result in the catgut embedding group was higher than that in the western medication group (P<0. 05). (4)After treatment, the SP level was lower than that before treatment in the two groups (both P<0. 05); the level of p-EP was increased as compared with that before treatment (both P<0. 05). The SP level in the catgut embedding group was lower than that in the western medication group (P<0. 05); the level of β-EP was higher than that in the western medication group (P<0. 05).</p><p><b>CONCLUSION</b>The acupoint catgut embedding therapy apparently relieves the clinical symptoms of chronic pelvic cavity pain syndrome differentiated as kidney deficiency and stagnation of damp heat as well as the condition of anxiety and depression, increases lecithin body numbers in prostatic fluid and β-EP level and reduces SP level.</p>

Effects and mechanisms of the inflammatory reaction related to NASH and induced by activation of the cholinergic anti-inflammatory pathway / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects and mechanisms of the inflammatory reaction related to nonalcoholic steatohepatitis (NASH) and induced by activation of the cholinergic anti-inflammatory pathway.</p><p><b>METHODS</b>A mouse model of NASH was established by feeding a high-fat and high-sugar diet.Activation of the cholinergic anti-inflammatory pathway was achieved by nicotine administration to the NASH modeled mice and normal controls. Liver biopsies were taken and the concentrations of cytokines were measured. Isolated liver primary Kupffer cells and RAw264.7 cells were cultured, pre-treated or not with lipopolysaccharide (LPS) and exposed to nicotine, after which the supernatant concentrations of IL-6 and TNFa were determined by ELISA. The protein expression levels of phosphorylated (p)-NF-kB and I k B were detected in primary cultured Kupffer cells by western blotting.</p><p><b>RESULTS</b>The mouse model of NASH was successfully established, as evidenced by findings from liver biopsy and serum liver function tests. The degree of liver inflammation in the NASH mice decreased after nicotine administration, and the level of serum TNFa also significantly decreased. The levels of serum TNFa were 21.95+/-0.8 pg/mL in nicotine-treated mice and 38.07+/-1.7 pg/mL in the non-nicotine-treated NASH mice (P less than 0.05). The nicotine treatment also significantly reduced the concentration of TNFa in the culture supernatants of Kupffer cells after LPS stimulation; moreover, the supernatant level of TNFa decreased significantly after the nicotine treatment (Pless than 0.05). LPS stimulation of the RAw264.7 cells led to an increased level ofp-NF-kB and a reduced level ofI-kB, suggesting that the NF-kB pathway had been activated; different doses of nicotine pre-treatment led to down-regulation of the p-NF-kB level and up-regulation of the I-kB level, both in dose-dependent manners.</p><p><b>CONCLUSION</b>Activating the cholinergic anti-inflammatory pathway inhibits the NASH-related inflammatory reaction, and the mechanism for this inhibition involves the NF-kB signaling pathway.</p>

Clinical efficacy of combined procedures for the surgical correction of congenital ptosis accompanied by epicanthus / 中华医学美学美容杂志

Objective To observe the effects of combined surgeries for the correction of congenital ptosis accompanied by epicanthus.Methods 80 cases (105 eyes) of congenital ptosis were analyzed in this study.64 cases (80 eyes),with levator muscle strength more than 4 mm,were treated with suspension of frontal aponeurosis,and 16 cases (25 eyes),with levator muscle strength less than 4 mm,were treated with shorten operation of levator palpebrae superioris.Furthermore,all these patients were corrected the epicanthus at the same time.Results Except the under correction in 5 cases (8 eyes),the remaining patients were corrected satisfactorily.Postoperative palperbal fissure length was averagely increased by 2.0± 1.0 mm,inner canthic diameter was averagely decreased by 4.0± 2.0 mm,the eyelid height was increased by average 9.0±1.0 mm,the inclination of palpebral fissure was 12.0°±2.0°;64 cases (80 eyes) were treated with suspension of frontal aponeurosis,postoperative recurrence were 7.5％ (6/80),no postoperative recurrence were 92.5％ (74/80),complication were 5.0％ (4/80);16 cases (25 eyes) were treated with shorten operation of levator palpebrae superioris,postoperative recurrence were 8.0％ (2/25),and no postoperative recurrence were 8.0％ (2/25);complication rates were 4.0％ (1/25).Conclusions This procedure of combined surgeries can reduce the recurrence rate effectively.Moreover,the satisfactory,functional,and aesthetic results can be obtained simultaneously with this method.

Study on Standard Revision of Portulaca grandiflora / 中国药房

OBJECTIVE:To improve the quality standard of Portulaca grandiflora. METHODS:TLC was used to identify the P. grandiflora with the reference of letuolin and apigenin. HPLC was performed on the column of Hanbon-C18 with the mobile phase of methanol-acid gradient[water-acetic acid(61∶4)] at the flow rate of 1.0 ml/min,the detection wavelength was 335 nm,the temperature was 35 ℃ and the volume was 10 μl. RESULTS:There was a good linear relationship in the quality concentration of scutellarin and luteolin respectively in th range of 0.299 5-1.872 0 μg(r=0.999 8)and 0.015 1-0.094 4 μg(r=0.999 5);the RSDs of precision,stability and reproducibility tests were ≤2.61%. The average recovery was respectively 100.42%(RSD=0.81%,n=6) and 100.65%(RSD=1.64%,n=6). CONCLUSIONS:The established TLC and HPLC are stable,reliable,simple and can be used for the qualitative and quantitative analysis of P. grandiflora.

Effect and mechanism of rat macrophage α7-acetylcholine receptor mediated anti-inflammatory ;pathway on endotoxin-induced inflammation reaction / 中华临床感染病杂志

Objective To investigate the effect of rat macrophage α7-acetylcholine receptor (α7-AChR )-mediated cholinergic anti-inflammatory pathway on endotoxin-induced inflammation reaction . Methods Density gradient centrifugation method was used to isolate rat primary macrophages and flow cytometry was used to identify the cell purity .α7-AChR in macrophages was detected by fluorescence confocal microscopy and Western blot .After 1ipopolysaccharides ( LPS) was added to the culture media of primary culture of macrophages , the concentration of tumor necrosis factor ( TNF)-αin the supernatant was determined by enzyme linked immunosorbent assay (ELISA).The expression of p-NF-κB protein in primary cultured macrophages was detected by Western blot .ANOVA was used to analyze TNF-αlevels after adding different concentrations of nicotine .Results α7-AChR was observed by fluorescence confocal microscope in primary macrophages .Nicotine could significantly reduce the concentration of TNF-αin culture supernatants of macrophages after LPS stimulation .When the concentrations of nicotine were 0, 1, 10, 100μmol/L, the concentrations of TNF-αwere (2 123 ±86), (1 486 ±80), (1 316 ±83) and (1 090 ±77)pg/mL, respectively (t=16.33, 20.18 and 26.83, P<0.05).The level of p-NF-κB in macrophages was also reduced when nicotine added .Conclusion Activation of macrophage α7-AChR can inhibit the endotoxin-induced release of inflammatory factor TNF-α, which may be through NF-κB signal pathway .

The clinical observation of fluticasone salmeterol inhaler combined with salbutamol inhalation of oxygen-driven treatment of chronic obstructive pulmonary disease with acute exacerbation / 中国基层医药

Objective To observe the clinical efficacy of fluticasone salmeterol inhaler combined with salbutsmol inhalation of oxygen-driven treatment of chronic obstructive pulmonary disease with acute exacerbation.Methods 60 patients of chronic obstructive pulmonary disease with acute exacerbation,randomly divided into observation and control groups of 30 patients each.The control group on the basis of conventional treatment with salmeterol plus fluticasone inhalation powder compound and the oxygen-driven albuterol inhalation aerosol liquid,the other group by conventional therapy based on the use of oxygen-driven albuterol inhalation aerosol solution,then compare the clinical curative effects.Results There were 18 examples excellencely,10 examples improve,2 examples ineffective of observation group with total effective rate 93.3％ ; 9 examples excellencely,11 examples improve,10 examples ineffective of control group with total effective rate 66.7％.The contrast of total effective power with the two groups exists the statistical significance ( x2 =4.60,P ＜ 0.05 ) . In the observation group,FEV1 and FVC of post-treatment,were better than pretherapy and post-treatment of control group(t =2.629,2.649,2.064,2.051,P ＜0.05).Conclusion Salmeterol and fluticasone propionate inhalation powder hybrid atomizing inhalation of salbutamol effective than salbutamol alone.

Measurement and clinical significance of IL-8 、IL-13 and IL-18 in serum of patients with bronchial asthma / 中国基层医药

ObjectiveTo investigate the change and significance of IL-8,IL-13 and IL-18 level in serum of patients with bronchial asthma.Methods45 patients with acute exacerbation of bronchial asthma were selected (acute group),and remission group 45 patients and 30 healthy persons (control group) were selected.The serum IL-8,IL-13 and IL-18 levels were measured respectively by ELISA.ResultsThe serum IL-8,IL-13 and IL-18 levels in acute group were significantly higher than that in control group and remission group ( t =5.21,5.13,4.99,5.32,5.48,5.59,P ＜ 0.05 ) ; The serum IL-8,IL-13 and IL-18 levels in remission group were significantly higher than that in control group(t =4.18,4.71,4.89,P ＜0.05).ConclusionThe levels of serum IL-8,IL-13 and IL-18 may be related to the severity of asthma attack.IL-8,IL-13 and IL-18 played important role in both pathogenesis and determination of asthma.

The expression and significance of protein tyrosine phosphatase SHP2 in juvenile rat bacterial meningitis model / 中华传染病杂志

Objective To explore the expression and significance of protein tyrosine phosphatase SHP2 in experimental bacterial meningitis.Methods A juvenile rat bacterial meningitis model was established by direct intraeisternal injection with Streptococcus pneumoniae. Uninfected control animals were mock-infected with sterile saline.The transcription and expression of SHP2 were detected by reverse transcription-polymerase chain reaction(RT-PCR),Western blot and immunohistochemistry techniques respectively at 1,3,7 and 14 days after infection.White blood cell(WBC)count,concentration of tumor necrosis factor-alpha(TNF-α)in cerebrospinal fluid(CSF)were also measured.Variables that were not normally distributed were compared by the Kruskal-Wallis test.Multiple comparison used Student-Newman-Keuls(SNK).The association between variables was assessed by using the Pearson correlation coefficient.Results Compared with uninfected controls,rats with bacterial meningitis showed a significant upregulation of SHP2 at both mRNA and protein levels(F=12.74,P<0.01;F=198,P<0.01).S HP2 mRNA levels peaked at 3 days after infection increasing more than five fold and remained at high levels at 7 days.In parallel,SHP2 protein levels began to increase at 3 days after infection,reaching a maximal increase of nearly nine fold at 7 days and remained at high levels at 14 days. Immunohistochemical analysis of SHP2 expression in the juvenile rat brain demonstrated that SHP2 labelling cells,identified as neuronal and glial cells,widely distributed in the cerebral cortex and the increased immunoreactive cells around the third ventricle were mainly glial cells.In addition,the protein levels of SHP2 and WBC counts were significantly correlated with each other(r=0.77,P<0.01),while there were no significant correlations between levels of SHP2 and TNF-a (r=0.08,P>0.05).Conclusions SHP2 may participate the pathological progress of the bacterial meningitis,restrating the inflammation and accelerating the renovation,so it can be regard as an index to measure the state of the illness.