Upregulation of KLF14 expression attenuates kidney fibrosis by inducing PPARα-mediated fatty acid oxidation.

Tubulointerstitial fibrosis (TIF) is essential during the development of end-stage kidney disease (ESKD) and is associated with the impairment of fatty acid oxidation (FAO). Kruppel-like factor 14 (KLF14) is an important gene in lipid metabolism, but its role in TIF remains unknown. TGF-ß-stimulated HK-2 cells and mouse unilateral ureteral obstruction (UUO) were used as renal fibrosis models. The role of KLF14 in the process of renal fibrosis was verified by gene knockout mice, genetic or pharmacological interference in animal model and cell model respectively. In the current study, we found that KLF14 expression increased after activation of the TGF-ß signaling pathway during TIF. In KLF14-/- mice, more severe fibrosis was observed after unilateral ureteral obstruction (UUO) was induced. In human HK2 cells, knockdown of KLF14 led to more severe fibrosis induced by TGF-ß1, while overexpression of KLF14 partially attenuated this process. Specifically, KLF14 deficiency decreased mitochondrial FAO activity, resulting in lipid accumulation. Thus, the energy supply to the cells was insufficient, finally resulting in TIF. We further proved that KLF14 could target peroxisome proliferator activated receptor alpha (PPARα) as a transcriptional activator. This study identified the upregulation of KLF14 expression in response to kidney stress during the process of fibrosis. Upon TIF, the activated TGF-ß signaling pathway can enhance KLF14 expression, while the upregulation of KLF14 expression can decrease the degree of TIF by improving FAO activity in tubular epithelial cells and recovering the energy supply mediated by PPARα.

Acyl-CoA Thioesterase 8 and 11 as Novel Biomarkers for Clear Cell Renal Cell Carcinoma.

BACKGROUND: Clear cell renal cell carcinoma (ccRCC) is essentially a metabolic disorder characterized by reprogramming of several metabolic pathways. Acyl-coenzyme A thioesterases (ACOTs) are critical enzymes involved in fatty acid metabolism; however, the roles of ACOTs in ccRCC remain unclear. This study explored ACOTs expressions and their diagnostic and prognostic values in ccRCC. METHODS: Three online ccRCC datasets from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) were utilized to measure the expressions of ACOTs in paired normal and tumor tissues. Receiver operating characteristic (ROC) curves were depicted to assess the diagnostic values of ACOTs in ccRCC. Quantitative real-time PCR and immunohistochemical analysis were performed to validate the ACOT11 expression in ccRCC cell lines and clinical samples. Survival curves and Cox regression analysis were used to evaluate the predictive values of ACOTs in clinical outcome of ccRCC patients. Functional enrichment analyses and correlation analysis were carried out to predict the potential roles of ACOT8 in tumorigenesis and progression of ccRCC. RESULTS: ACOT1/2/8/11/13 were found to be significantly downregulated in ccRCC samples. In particular, ACOT11 was decreased in almost every matched normal-tumor pair, and had extremely high diagnostic value as shown by ROC curve analysis (AUC = 0.964). The expression of ACOT11 was further verified in ccRCC cell lines and clinical samples at mRNA and protein levels. Furthermore, clinical correlation analysis and survival analysis indicated that ACOT8 was correlated with disease progression and was an independent predictor of unfavorable outcome in ccRCC. Moreover, functional analyses suggested potential roles of ACOT8 in the regulation of oxidative phosphorylation (OXPHOS), and correlation analysis revealed an association between ACOT8 and ferroptosis-related genes in ccRCC. CONCLUSION: Our study revealed that ACOT11 and ACOT8 are promising biomarkers for diagnosis and prognosis of ccRCC, respectively, and ACOT8 may affect ccRCC development and progression through the regulation of OXPHOS and ferroptosis. These findings may provide new strategies for precise diagnosis and personalized therapy of ccRCC.

Is the Preoperative Level of Procalcitonin a Valid Indicator for Predicting Postoperative Fever After Percutaneous Nephrolithotomy?

OBJECTIVE: To evaluate the risk factors for postoperative fever and to identify the value of preoperative procalcitonin (PCT) in predicting postoperative fever after percutaneous nephrolithotomy (PNL). PATIENTS AND METHODS: Patients who underwent PNL between January 2014 and March 2017 were studied. In total, 363 medical records with complete data were determined to be eligible for analysis. Patients were classified into a control or febrile group according to the presence of a body temperature over 38°C. Demographic and perioperative data were compared between the groups. Variables found to be statistically significant were included in a binary logistic regression analysis. RESULTS: Ninety-one (25.1%) patients experienced postoperative fever. Univariate analysis revealed a statistically significant difference between postoperative fever and factors, such as sex (p = 0.009), preoperative fever (p < 0.001), stone burden (p < 0.001), pyuria (p = 0.013), urine culture (p < 0.001), and serum levels of C-reactive protein (CRP) (p = 0.003), PCT (p < 0.001), and interleukin-6 (IL-6) (p = 0.003). Binary logistic regression analysis indicated the presence of preoperative fever (p = 0.037), stone burden >353 mm2 (p = 0.002), PCT >0.05 ng/mL (p < 0.001), or positive urine culture (p = 0.004) as independent risk factors for postoperative fever following PNL. CONCLUSIONS: We concluded that patients with preoperative fever, stone burden >353 mm2, PCT >0.05 ng/mL, or positive urine culture were more likely to develop postoperative fever and that routinely detecting PCT levels before PNL would be helpful in predicting postoperative fever.

Relaxin abrogates renal interstitial fibrosis by regulating macrophage polarization via inhibition of Toll-like receptor 4 signaling.

Renal fibrosis is a common feature of chronic kidney disease (CKD). To inhibit the CKD process, it is important to prevent renal fibrosis, though CKD remains incurable. Renal fibrosis can be inhibited by relaxin in several experimental models, but the mechanism of relaxin for antifibrotic potential is still not clear. And here we have studied the role of relaxin in macrophage polarization and renal inflammation after unilateral ureteral obstruction (UUO). Our results show that relaxin can downregulate the Toll-like receptor (TLR) 4 signaling, shift macrophage polarization toward the M2 phenotype and ameliorat renal fibrosis in the early stages of UUO. In vitro experiments, it has been confirmed that relaxin can downregulate the TLR4 signaling and induce the M2 macrophage transition. Furthermore, the transitional actions of macrophage phenotype induced by relaxin are significantly blocked by TAK-242, a TLR4 antagonist, in vitro experiments. Thus, there is a novel mechanism of relaxin for antifibrosis that shifts macrophage polarization toward the M2 phenotype via inhibition of TLR4 signaling.

EZH2 mediates lidamycin-induced cellular senescence through regulating p21 expression in human colon cancer cells.

Lidamycin (LDM) is a novel member of the enediyne antibiotics identified in China with potent antitumor activity. However, it remains unclear whether LDM has potential molecular targets that may affect its antitumor activity. Enhancer of zeste homolog 2 (EZH2) functions as a histone lysine methyltransferase and mediates trimethylation on histone 3 lysine 27 (H3K27me3). High EZH2 level is found to be positively correlated with the aggressiveness, metastasis and poor prognosis of cancer. Here, we aim to study the role of EZH2 in LDM-induced senescence, as well as in the cytotoxicity of LDM in human colon cancer cells. LDM is found to be relatively more potent in inhibiting the colon cancer cells harboring high EZH2 level and induces irreversible cellular senescence at IC50 dose range, as evidenced by senescence-associated ß-galactosidase staining, cell cycle arrest and molecular changes of senescence regulators including p21 in HCT116 and SW620 cells. More importantly, LDM is found to markedly inhibit EZH2 expression at both protein and mRNA levels upon the induction of p21 and cellular senescence. LDM also selectively inhibits EZH2 expression as compared with other histone lysine methyltransferases. Knockdown of p21 with siRNAs abolishes LDM-induced senescence, whereas EZH2 knockdown markedly increases p21 expression and causes senescent phenotype. Enrichment of both EZH2 and H3K27me3 levels in the p21 promoter region is reduced by LDM. Moreover, EZH2 overexpression reduces cellular senescence, p21 expression and DNA damage response upon LDM exposure. LDM also demonstrates potent antitumor efficacy in xenografted animal models. Collectively, our work provides first demonstration that EZH2 may mediate, at least partially, the senescence-inducing effects of LDM by regulating p21 expression and DNA damage effect. Thus, EZH2 may serve as a potential target and biomarker to indicate the clinical efficacy of the potent enediyne antitumor drug.

Design, synthesis and biological evaluation of novel benzimidazole-2-substituted phenyl or pyridine propyl ketene derivatives as antitumour agents.

A series of novel benzimidazole-2-subsituted phenyl or pyridine propyl ketene derivatives were designed and synthesized. The biological activities of these derivatives were then evaluated as potential antitumour agents. These compounds were assayed for growth-inhibitory activity against HCT116, MCF-7 and HepG2 cell lines in vitro. The IC50 values of compounds A1 and A7 against the cancer cells were 0.06-3.64 µM and 0.04-9.80 µM, respectively. Their antiproliferative activities were significantly better than that of 5-Fluorouracil (IC50: 56.96-174.50 µM) and were close to that of Paclitaxel (IC50: 0.026-1.53 µM). The activity of these derivatives was over 100 times more effective than other reported structures of chalcone analogues (licochalcone A). A preliminary mechanistic study suggested that these compounds inhibit p53-MDM2 binding. Compounds A1, A7 and A9 effectively inhibited tumour growth in BALB/c mice with colon carcinoma HCT116 cells. The group administered 200 mg/kg of compound A7 showed a 74.6% tumour growth inhibition with no signs of toxicity at high doses that was similar to the inhibition achieved with the 12.5 mg/kg irinotecan positive control (70.2%). Therefore, this class of benzimidazole-2-subsituted phenyl or pyridine propyl ketene derivatives represents a promising lead structure for the development of possible p53-MDM2 inhibitors as new antitumour agents.

Effect of heat shock pretreatment on apoptosis and metallothionein expression in rat cardiomyocytes.

To investigate the effect of heat shock pretreatment on apoptosis and mitochondrial metallothionein (MT) expression in rat cardiomyocytes. In vitro cultured H9C2 cells were randomly divided into three groups: control, hydrogen peroxide (H2O2) injury, and H2O2 injury after heat shock pretreatment (n = 6 per group). Cardiomyocyte apoptosis and caspase-3 activity were assayed after treatment. Mitochondrial cytochrome (cyt) c and MT expression was assayed by Western blotting. Compared with the control group, the H2O2 injury group had a growing number of apoptotic cardiomyocytes (P < 0.01) and significantly elevated caspase-3 activity (P < 0.01) with markedly increased mitochondrial cyt c and MT expression (P < 0.01). After heat shock pretreatment, the numbers of apoptotic and necrotic cardiomyocytes (P < 0.01) and the caspase-3 activity significantly declined (P < 0.01), while mitochondrial cyt c and MT expression continued to increase (P < 0.01) compared with the H2O2 injury group. Heat shock pretreatment inhibits cardiomyocyte apoptosis, which may have a protective effect on cardiomyocytes by increasing the expression of myocardial protective MT and reducing the release of mitochondrial cyt c.

[A survey on respiratory infectious diseases related health literacy and health behavior in Fengtai district, Beijing].

OBJECTIVE: This study aims to survey respiratory infectious disease related health literacy (RIDHL) and health behavior (RIDHB) among residents in Fengtai district, Beijing, analyze impact factors of RIDHL , explore the association between RIDHL and RIDHB. METHODS: Multistage sampling was employed and 1100 respondents were surveyed by self-designed questionnaires, which including social-demographic characteristics and evaluation of RIDHL and RIDHB. The survey results were described, the impact factors of RIDHL and the association between RIDHL and RIDHB were analyzed by analysis of variance or covariance. RESULTS: A total of 998 qualified questionnaires were recollected with the effective rate of 90.7%. The respondents aged from 15 to 65, scored (71.3 +/- 19.0) points in RIDHL test. Of those respondents, 25.7% (256/998), 43.2% (432/998) and 31.1% (310/998) were evaluated as low( <60 points), medium (60 - 85 points), and high level ( > 85 points) of RIDHL, respectively. There were significant difference in RIDHL scores between registered and non-registered residents, who scored (74.1 +/- 18.9) and (68.4 +/- 18.8) points, respectively (P < 0.01). RIDHL sections were ranked as audiovisual (77.6%, 4647/5988), internet using (75.2%, 2251/2994), reading (74.6%, 3724/4990), map using (68.3%, 4090/5988) and quantitative (65.5%, 5230/7984) according to the accurate rates from high to low. Analysis of variance or covariance showed that RIDHL scores were significantly different among respondents with different ages, nationalities, educational levels, occupations, and incomes (P < 0.01), yet no significant differences among those with different genders and marital status (P > 0.05). Respondents scored (69.7 +/- 15.5) points in RIDHB test. The RIDHB scores ((64.5 +/- 15.0), (70.4 +/- 15.6), (72.5 +/- 14.9) points, respectively) increased among residents with low, medium and high level of RIDHL (P < 0.01). CONCLUSION: Residents in Fengtai district, Beijing possessed medium level of RIDHL. The non-registered residents showed lower RIDHL than registered residents. Ages, nationalities, educational levels, occupations, and incomes were impact factors of RIDHL. People with higher level of RIDHL also showed a higher level of RIDHB.

[Bone setting manipulation combined with surgery for the treatment of calcaneal fractures].

OBJECTIVE: To investigate the clinical effects of bone setting manipulation on simplified operation and swelling reduction in treating calcaneal fractures. METHODS: From December 2005 to November 2010, 42 patients with calcaneal were reviewed, including 25 males and 17 females, ranging in age from 18 to 74 years, with an average of 41.4 years. Twenty patients had fractures in the left and 22 in the right. Twenty-three patients were treated with anatomical plate fixation, 19 patients were treated with Kirchners wires or cannulated screws fixation. The average period of swelling in soft tissue, joint function and complications were evaluated. RESULTS: Forty-two patients were followed up, and the duration ranged from 3 to 18 months, with a mean of 8.3 months. The pain was markedly relieved at the next day after reduction, and the swelling was relieved in 3 to 5 days. The operative was simplified and the average operative time was 90 minutes. The swelling was relieved in 4 to 7 days after the operation, and the necrosis of skin was not found. The average postoperatively Böhler angle was (31 +/- 3.2) degrees. Gissane angle was (112 +/- 5.3) degrees. Calcaneal width was (30.2 +/- 0.89) mm. According to Maryland foot function score system, 16 patients got an excellent result, 18 good, 6 fair and 2 bad. CONCLUSION: The operation is simplified, and skin complications decrease, as well as the detumescence period is shortened.

[Karyotype analysis of Picris davurica chromosome].

OBJECTIVE: To systematically study Picris davurica Fisch chromosome karyotype. METHODS: Sections combined with micrograph were used to analyse chromosome. RESULTS: Chromosome number of normal diploid was 2n = 10, karyotype formula was K (2n) = 10 = 2 m + 8 sm, the relative length was 2n = 10 = 2 L + 2 M2 + 4 M1 + 2 S, which belonged to "3 A" type. The total length of chromosome groups was 12.51 microns, the total length of long arms was 8.36 microns, the AS. K% was 66.83%. The total volume of chromosome was 11.71 microns3. CONCLUSION: Picris davurica chromosome number and morphological character were clear.

[Karyotype analysis of Astragalus complanatus chromosome].

Micrograph were used to analyse the number, karyotype and volume of chromosome of Astragalus complanatus. The normal diploid 2n = 16, karyotype formula based on Levean publication (1964) was K(2n) = 16 = 10 m + 6 sm. According to the method of S.R. Guo, the chromosome relative length was 2n = 16 = 6M2 + 10M1, which belonged to "2A" type according to the Stebbins' karyotype classification. The total length of chromosome groups was 33.05 microns, total length of long arms was 19.76 microns, from Arano's method the AS.K% was 59.79%. The total volume of chromosome was 50.6 micron 3.