Global burden of ischemic heart disease associated with high red and processed meat consumption: an analysis of 204 countries and territories between 1990 and 2019.

BACKGROUND: Multiple studies have indicated an association between red and processed meat consumption and the incidence of ischemic heart disease (IHD). In this study, we aimed to assess the burden of IHD caused by a diet high in red and processed meat in 204 countries and territories between 1990 and 2019, using data from the Global Burden of Disease (GBD) 2019. METHODS: We extracted data from the GBD 2019, which included the number of deaths, age-standardized mortality rates (ASMR), disability-adjusted life years (DALYs), and age-standardized DALYs rates (ASDR) attributed to IHD caused by a diet high in red and processed meat. We then calculated the burden of IHD attributable to a high intake of red and processed meat in each country and territory, stratified by age, sex, and socio-demographic index (SDI). RESULTS: Globally, a high intake of red meat was responsible for 351,200 (95% uncertainty interval (UI): 559,000-642,700) deaths from IHD in 2019, while a high intake of processed meat was associated with 171,700 (95% UI: 30,100-320,000) deaths from IHD. Between 1990 and 2019, while the corresponding age-standardized rates declined, the numbers of deaths and DALYs increased. China had the highest number of deaths [98,386.9 (95% UI: 14,999.3-189,812.7)] caused by a high intake of red meat, while United States of America [33,129.6 (95% UI: 7,150-59,593.8)] was associated with the highest number of deaths caused by high intake of processed meat for IHD in 2019. Males experienced a greater burden of IHD caused by a high intake of red and processed meat than females. The ASMR and ASDR of IHD attributed to a high intake of red meat decreased in countries with high SDI, high-middle SDI and low SDI, while the ASMR and ASDR of IHD attributed to a high intake of processed meat decreased only in countries with high SDI and high-middle SDI. CONCLUSION: Although there is a decline in the ASMR and ASDR of IHD caused by a high intake of red and processed meat, there is also an increase in deaths and DALYs number globally. Additionally, there is a heterogeneous burden of IHD related to a high intake of red and processed meat across regions and countries, with males experiencing a greater burden than females. Implementing targeted policies and interventions is required to reduce the burden of IHD caused by a high intake of red and processed meat.

Δ133p53 decreases the chemosensitivity of carcinoma cell line H1299.

The research evaluated the effect of Δ133p53 on the chemosensitivity of lung adenocarcinoma cell line H1299. By this study, the drug-resistant molecular marker and a new target for cancer therapy could be provided. Δ133p53 or negative control plasmid were transferred into H1299 cells by lentivirus vector. The expression of Δ133p53 in transfected cells was examined using immunofluorescence. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method and colony formation test were applied to detect drug sensitivity after cisplatin or 5-fluorouracil (5-FU) treatment. After cisplatin (CDDP)/FU treatment, MTT assay demonstrated that the inhibition rate of H1299/Δ133p53 cell was reduced compared with that of the H1299 and H1299/NEG cells at the same concentration of drug. The 50% inhibitory concentrations (IC 50 ) of CDDP and 5-FU rose by 36.1 and 30.2%, respectively (P < 0.05). The colony formation assay suggested that the cell proliferation ability of H1299/Δ133p53 cell was prominently increased when compared with that of control group H1299 and H1299 /NEG cells (P < 0.05). The present study demonstrated that the transfection of the Δ133p53 gene in H1299 cells led to the reduction of chemosensitivity.

Effect of the p53α gene on the chemosensitivity of the H1299 human lung adenocarcinoma cell line.

To investigate the effects of tumor protein p53 (p53 or TP53) α gene on the chemosensitivity of the H1299 human lung adenocarcinoma cell line, the recombinant vector pEGFP-p53α was constructed. The vector pEGFP-p53α was transfected into the cultured p53-null H1299 cells using Lipofectamine 2000. The G418-resistant cells were then selected. The expression of the p53α gene in these cells was examined using reverse transcription-polymerase chain reaction, and TP53 protein expression was examined using western blot analysis and immunocytochemistry. An MTT assay and colony formation assay were used to analyze the response of the transfected cells to cisplatin (CDDP). DAPI staining was used to determine the level of apoptosis of the transfected cells. The transfected H1299 human lung adenocarcinoma cells stably expressed TP53 protein. The MTT assay demonstrated that the 50% inhibitory concentrations for the H1299, H1299/pEGFP-N1 and H1299/pEGFP-p53α cells were 28, 24 and 18 µmol/l, respectively. The survival rate of H1299/pEGFP-p53α cells was significantly reduced compared with that of H1299 and H1299/pEGFP-N1 cells (P<0.05). The colony formation assay and DAPI staining identified that the colony formation rate and the number of apoptotic cells of H1299/pEGFP-p53α were significantly reduced, compared with those of the H1299 and H1299/pEGFP-N1 cells (P<0.05). Therefor, the present study demonstrated that the transfection of H1299 cells with the p53α gene resulted in an increase in sensitivity to CDDP chemotherapy. The combination of CDDP and gene therapy for H1299 lung adenocarcinoma cell line provides an experimental basis for clinical research.

Effect of combined 5-aza-2'deoxycytidine and cisplatin treatment on the P15 lung adenocarcinoma cell line.

Aberrant promoter hypermethylation resulting in the epigenetic silencing of apoptosis-associated genes is a key process in the chemotherapeutic treatment of cancer. The nucleoside analog, 5-aza-2'deoxycytidine (DAC), inhibits the activity of DNA methyltransferase enzymes and is able to restore the expression levels of genes that have been silenced by aberrant DNA methylation. The aim of the present study was to investigate the effect of combined treatment with DAC and cisplatin (CDDP) on the lung adenocarcinoma cell line, P15. Growth inhibition was examined using a clone formation assay and growth inhibitory activities by cell counting during treatment with DAC alone, CDDP alone or DAC followed by CDDP. In addition, changes in the mRNA expression levels of various apoptosis-associated genes following treatment with increasing concentrations of DAC were determined using reverse transcription-polymerase chain reaction. Furthermore, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) analysis was used to detect the number of apoptotic P15 tumor cells following treatment with DAC and/or CDDP. The results indicated that DAC treatment alone restored the mRNA expression levels of p73, p16INK4a , B-cell lymphoma (Bcl)-2-associated agonist of cell death and Bcl-2-associated X protein. In addition, combined therapy with DAC and CDDP was found to significantly suppress the growth of P15 tumor cells compared with DAC or CDDP treatment alone. In conclusion, DAC may enhance the chemosensitivity of the P15 cell line to treatment with CDDP.

[Effect and significance of BH3-only protein in targeted therapy of
 non-small cell lung cancer].

Lung cancer is the leading cause of cancer deaths throughout the world. Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. In traditional anti-cancer therapy, promotion of apoptosis in NSCLC is an important part of treatment, but anticancer drugs have the toxic side effects, resistance and other problems. Therefore, the search for new targets of anticancer drugs becomes one of the foci in the treatment of NSCLC. The BH3-only protein plays an important role in activation and communication in apoptosis pathways. BIM is the core member in BH3-only protein family. The target at BIM in the treatment of NSCLC has an irreplaceable role. This paper briefly describes the BCL-2 family and BH3-only pro-apoptotic protein, elaborates the important role of BIM and BH3-only protein in targeted therapy of NSCLC.

Novel mechanism of intra­renal angiotensin II-induced sodium/proton exchanger 3 expression by losartan in spontaneously hypertensive rats.

The present study aimed to investigate the molecular pharmacodynamic mechanisms of losartan used in the treatment of hypertension. A total of 12 spontaneously hypertensive rats (SHR) were divided randomly into an SHR group treated with saline and LOS group treated with losartan. Six Wistar­kyoto rats (WKY) were enrolled as the WKY group with saline in the study. The LOS group received 30 mg/kg/day losartan by intragastric injection, while the SHR and WKY were fed the same volume of saline. The dosage was modulated according to the weekly weight. Changes in blood pressure were measured by the indirect tail cuff method. Angiotensin (Ang) II production in the plasma and renal tissue was measured by an immunoradiometric method. Na+/H+ exchanger (NHE)3 and serum and glucocorticoid­inducible kinase (SGK)1 were assessed by quantitative polymerase chain reaction (qPCR) and western blot analysis. When compared with the WKY group, the blood pressure of the SHR and LOS groups were higher prior to treatment with losartan. Following two weeks, blood pressure was reduced and the trend continued to decrease over the following six weeks. The plasma and renal tissue levels of Ang II in the SHR and LOS groups were significantly higher than those in the WKY group. NHE3 and SGK1 were increased at the mRNA and protein level in the SHR group, and losartan reduced the expression of both of them. The results suggested that in hypertensive rats, the circular and tissue renin angiotensin systems were activated, and the increased Ang II stimulated the expression of NHE3 and SGK1, which was reduced by losartan. Therefore, the effects of losartan in hypertension may be associated with the Ang II­SGK1­NHE3 of intra­renal tissue.

[P53 family proteins provide new insights into lung carcinogenesis and clinical treatment].

The transcription activation of p53 plays an important role in the maintenance of genetic stability. P53 is an intensive study tumor suppressor, which has been called the "gene guider". The p53 family members p63, p73 have high homologous sequence with p53. Some of them can bind to the p53-responsive genes and transcript the downstream genes. Human lung cancer is one of the most common malignant tumors in the world. Abnormality of the p53 gene is the significant event in lung cancers, which leads to the poor prognosis and the resistance of chemotherapy. A deep understanding of the relationship between p53 family members and lung cancers can provide a more reasonably targeted clinical approach. This paper will focus on the special function of p53 family members in the development, chemosensitivity and target treatment of lung cancer.

p73 expression is associated with cellular chemosensitivity in human non-small cell lung cancer cell lines.

p73 is a member of the p53 tumor suppressor protein family and induces apoptosis in tumor cells that lack functional p53. It has been demonstrated that methylation of CpG islands in the promoter and exon 1 region may result in silencing of the p73 gene. The aim of this study was to investigate the correlation between p73 gene expression and chemosensitivity in non-small cell lung cancer (NSCLC) cell lines. The expression of the p73 transcript in six NSCLC cell lines was investigated by reverse transcription-polymerase chain reaction (RT-PCR). The methylation status in these cell lines was determined by methylation-specific PCR (MSP) analysis. An in vitro demethylation assay was conducted using the DNA methyltransferase inhibitor 5-aza-2-deoxycytidine (5-aza-dC). Restored expression of p73 in the human lung squamous cell carcinoma cell line C57, both at the mRNA and protein level, was investigated by RT-PCR and immunohistochemistry, respectively. A colony formation assay was used to measure the surviving fraction of the C57 cell line. Transcript silencing of the p73 gene in the six NSCLC cell lines was observed and related to aberrant methylation. The expression of the p73 transcript and protein in the C57 cell line was restored by 5-aza-dC. The surviving fraction for colony formation in C57 cells pre-treated with 5-aza-dC was 0.059±0.006, which was significantly different from that of the control group (0.12±0.008; P<0.05). Our data demonstrated a significant correlation between expression of p73 and cellular chemosensitivity in NSCLC.

[Neonatal intrahepatic cholestasis caused by citrin deficiency: a histopathologic study of 10 cases].

OBJECTIVE: To investigate the diagnostic value of histopathological changes in the liver of patients with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD). METHODS: Liver specimens from 10 cases of NICCD were evaluated by hematoxylin-eosin stain, histochemistry and immunohistochemistry (EnVision method). SLC25A13 mutation analysis was performed to correlate with histopathology. RESULTS: Most specimens showed varying degrees of fat deposition in hepatocytes, necrotic inflammation, cholestasis and fibrosis (so-called tetralogy). The combination of the above four histological changes was highly characteristic for NICCD. With the progression of the disease, hepatic fibrosis deteriorated and ultimately led to cirrhosis. CONCLUSIONS: NICCD should be suspected in the presence of cholestasis during infancy. A liver biopsy must be performed to rule out other liver diseases. The tetralogy of the hepatic histopathological changes has a highly diagnostic value for NICCD, which is also practical for accurately assessing the degree of inflammation and fibrosis, and similarly the progression of hepatic cirrhosis.

Association of p73 G4C14-to-A4T14 polymorphism at exon 2 with the response of human lung adenocarcinoma cell lines to chemotherapy.

In order to assess the effect of p73 gene polymorphism G4C14-A4T14 on cisplatin-based chemosensitivity of human lung adenocarcinoma cell lines, we examined the differences in biological character and drug sensitivity affected by cisplatin between human lung adenocarcinoma cell lines A549 and P15. The allelic expression of p73 in A549 and P15 was studied by Sty I polymorphism analysis. MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay was used to analyse the response of these two cell lines to cisplatin. The changes in the biological behaviour of the cells were observed by colony formation assay. The drug-induced apoptosis of cells was measured by Hoechst and TUNEL techniques. Homozygous allelic expression was demonstrated in the two cell lines. AT/AT genotype appeared in A549, GC/GC genotype was detected in P15. Although the colony formation number decreased with an increasing cisplatin dose (P<0.05), there was no significant difference in colony-formation rate in these two cell lines (P>0.05). MTT assay also determined that the 50% inhibitory concentration (IC50) for A549 and P15 was 8.9 and 11.6 micromol/l, respectively; the IC50 value did not differ significantly between A549 and P15 (P>0.05). The cell apoptosis induced by cisplatin was demonstrated in both A549 and P15. P73 G4C14-A4T14 polymorphisms at exon 2 existed in human NSCLC (non-small-cell lung cancer) cell lines. Our data in vitro suggest that p73 G4C14-A4T14 polymorphism has no significant relationship to the cisplatin-based chemosensitivity in human lung adenocarcinoma.

The relationship between transforming growth factor beta1 expression and cold ischemia injury of rat donor kidney.

OBJECTIVE: Cold ischemia injury represents an independent risk factor which favors chronic allograft nephropathy (CAN). In order to investigate the role of transforming growth factor-beta 1( (TGF-beta1) in the progression of CAN, we studied the relationship between the expression of TGF-beta1 and cold ischemia injury in the renal tubular epithelia of rat donor kidney. METHODS: A total of 24 Wistar rats were used in this study. In terms of the time of cryopreservation of donor kidney, the 24 Wistar rats were randomly divided into 3 groups: 0 h group(control group), 24 h group, 48 h group. A block removal of donor kidney with in situ perfusion of cooling HC-A preservation solution was adopted. The rat kidney was preserved 0 h, 24h and 48 h at 0-4 degrees C respectively. The morphologic changes of proximal tubular epithelial cells in different cryopreservation time were observed under light microscope and transmission electron microscope. The expression of TGF-beta1 mRNA and protein in proximal tubular epithelial cells of different cryopreservation time group were detected by in situ hybridization and immunohistochemistry analysis. RESULTS: 1. In 24 h group, part of the proximal tubular epithelial cells showed slight degeneration. In 48 h group, the proximal tubular epithelial cells demonstrated severe hydropic degeneration and part of the cells developed necrosis and effluxion. 2. Only a small amount of TGF-beta1 protein and mRNA were expressed in the renal tubular epithelial cells of 0 h group. The positive unit (PU) value of TGF-beta1 protein and mRNA were 6.37+/-2.77 and 5.29+/-2.15, respectively. As the cold ischemia time prolonged, the PU value of TGF-beta1 protein increased at 24 h group (10.20+/-3.27) and 48 h group (17.17+/-3.96) . The PU value of TGF-beta1 mRNA also increased at 24 h group (11.31+/-3.34) and 48 h group (19.01+/-3.53). There was the significant difference of TGF-beta1 protein PU value or mRNA PU value among these groups(P<0.05). CONCLUSION: There was the significant correlation between the expression of TGF-beta1 and the degree of cold ischemia injury. The results suggest that TGF-beta1 might play the key role in regeneration and reparation of renal tubular epithelial cell injury. The overexpression of TGF-beta1 might be one of the mechanisms that initiate chronic allograft nephropathy.

Loss of p73 expression in six non-small cell lung cancer cell lines is associated with 5'CpG island methylation.

P73 is located on chromosome 1p36, a region that is frequently deleted in neuroblastoma and other tumors. P73 has been postulated to be a candidate tumor suppressor and imprinted gene that shares significant homology with the p53 gene. To investigate the pattern of inactivation of this gene in human non-small cell lung cancers, we studied the six NSCLC cell lines to identify abnormal methylation in exon 1 and the allelic expression using StyI polymorphism analysis. We also examined the p73 gene expression in these six cell lines by reverse transcription-PCR as well as the expression of p73 protein in the five cell lines inducing tumors by immunohistochemistry. Homozygous allelic expression was demonstrated in all six cell lines and the GC/GC genotype was the predominant type. P73 was aberrantly methylated in all these six lung cancer cell lines. Complete loss of the p73 expression both at mRNA and the protein level was associated with the p73 methylation. Our results show that methylation of the p73 gene could be an important mechanism in silencing expression of this gene in human non-small cell lung cancers.

Assessment of chemosensitivity by monitoring bcl-2 transcript kinetics in acute myeloid leukemias.

Enforced bcl-2 gene expression suppresses apoptosis and confers resistance to anticancer drugs. Here we established a real time fluorescence PCR assay to analyze the association between the bcl-2 gene expression and clinical chemosensitivity in acute myeloid leukemia. Expression levels of the bcl-2 gene were measured and normalized by beta-actin, a housekeeping gene expressed as endogenous reference. By applying real time PCR to clinical samples, we observed that although the bcl-2/beta-actin ratio was not related to FAB subtypes, the changing data following remission induction therapy clearly reflected drug-sensitivity. These results suggest that RT-PCR assay monitored the efficacy of the chemotherapy by quantifying the bcl-2 gene transcript in AML.