Harnessing Deaminated DNA to Modulate mRNA Translation for Controlled and Sequential Protein Expression.

Messenger RNA (mRNA) offers transformative potential in vaccines and therapeutics for a range of intractable diseases. While considerable efforts have focused on enhancing protein expression levels to improve efficacy, comparatively little attention has been given to regulating the rate and timing of protein expression. Given that sudden antigen bursts can overstimulate immune responses and pose serious risks in susceptible individuals, precise control over translation kinetics is essential for safe and personalized mRNA therapies. Herein, we describe the use of “damaged” DNA to modulate translation rates of mRNAs. Hybridization of deoxyuridine‐containing DNA to the 5′‐end of mRNA inhibits translation initiation, which is subsequently displaced via base excision repair (BER), enabling controlled expression. DNA strand lengths determine the rate and onset of translation (e.g., a 52‐nt DNA induces a 20‐fold slower expression with a 200‐min delay). This also enables the sequential expression of multiple mRNAs from a single cocktail. This strategy requires no chemical modification of the mRNA and produces no toxic byproducts, but only recyclable DNA fragments—offering a broadly applicable and biocompatible adjuvant for controlled mRNA translation.

To investigate the protein and mRNA expression of osteopontin (OPN) in the lung cancer tissue and explore the roles thereof in the development and progression of lung cancer. Immunohistochemistry and in situ hybridization were used to detect the protein and mRNA expression of OPN in 57 specimens of lung cancer tissue, 30 specimens of inflammatory pseudotumors and 20 specimens of pulmonary bulla, all obtained during operation. The OPN protein expression rate of the lung cancer tissue was 57.9% (33/57) , significantly higher than that of the inflammatory pseudotumor (16.7%, 5/30, chi2 = 13.581, P = 0.000). The OPN mRNA expression rate in the lung cancer tissue was 71.9% (41/57), significantly higher than that of the inflammatory pseudotumor (30.0%, 9/30, chi2 = 14. 138, P = 0.000). All the 20 specimens of pulmonary bullae were negative in the expression of OPN, both at the protein and mRNA levels. The OPN protein and mRNA expression rates of the lung cancer tissues with lymph node metastasis were 71.1% (27/38) and 86.8% (33/38) respectively, both significantly higher than those of the lung cancer tissue without lymph node metastasis [31.6% (6/19) and 42.1% (8/19) respectively, chi2 = 6.558, P = 0.010, and chi2 = 10.438, P = 0.001]. The OPN protein and mRNA expression rates of the non-small lung cancer tissues were 68.1% (32/47) and 78.7% (37/47) respectively, both significantly higher than those of the small lung cancer tissues [10% (1/10) and 25% (4/10) respectively, chi2 = 11.412, P = 0.001, and chi2 = 6.124, P = 0.013]. The OPN protein expression was positively correlated with the OPN mRNA expression in the lung cancer tissues (r = 0.623, P = 0.001). The 57 patients with lung cancer after surgery were followed up for 28 (24-40) months, 8 of the 32 patients with both positive OPN protein and mRNA expression had recurrence and 13 patients had a distant metastasis, while only 1 of the 15 cases negative in both OPN protein and mRNA expression showed recurrence (chi2 = 14.258, P = 0.000). 12 patients died in the both positive expression group but no patient died in the both negative expression group (chi2 = 7.554, P = 0.006). Over-expressions of OPN protein and OPN mRNA are found in lung cancer tissues and their expressions are correlated to the prognosis and metastasis of lung cancer.

Objective To study the expression of special AT-rich sequence-binding protein 1(SATB1) gene and its relationship with SLC22A18 protein expression in astrocytoma.Methods Fifty-six patients with astrocytomas (12 with grade Ⅰ,13 with grade Ⅱ,15 with grade Ⅲ,and 16 with grade Ⅳ),performed surgical excision in our hospitals from September 2006 to June 2010 and from September 2003 to June 2006,were chosen in our study; another 10 brain tissues from patients performed decompression operation resulting from cerebral hernia were selected as the controls.RT-PCR and Western blotting were used to detect the mRNA and protein expressions of SATB1.The SLC22A18 protein expression was detected by immunohistochemical assay.The relations between SLC22A18expressions and SA TB1 levels,and these two and the degree of malignancy were analyzed.Results RT-PCR and Western blotting revealed that positive mRNA and protein expressions were noted in 35patients with astrocytomas; the mRNA and protein expression rate and value of SATB1 in the astrocytoma tissues were significantly different among different grades of tumors (P＜0.05); the higher the malignancy grade,the higher mRNA and protein expression rate and value ofSA TB1; the protein expression value of SA TB1 had a positive correlation with the malignancy grade of tumors (r=0.987,P=0.000).And a few expressions of SA TB1 mRNA and protein were found in the tissues of controls.Immunohistochemical assay indicated that positive protein expression of SLC22A18 was noted in 19 astrocytoma tissues,and the protein expression rate of SLC22A18 in the astrocytoma tissues was significantly different among different grades of tumors (P＜0.05); the higher the malignancy grade,the lower expression of SLC22A18.And the protein expression of SLC22A18 was found in all the tissues of controls.The SATB1 expression rate in the tissues with negative SLC22A18 expression (81.1％) was significantly higher than that in the tissues with positive SLC22A18 expression (26.3％,P＜0.05).Conclusion SATB1 expresses in the astrocytoma tissues,indicating that it may play an important role in the pathogenesis of astrocytoma;up-regulation of SATB1 expression and dysfunction of SLC22A18 may play synergetic roles in the process of carcinogenesis of astrocytoma. Key words: Astrocytoma; Special AT-rich sequence binding protein 1; SLC22A18; Gene expression

To investigate the expression and clinical significance of mismatch repair genes hMLH1 and hMSH2 in sporadic colorectal carcinoma tissues. The expression of hMLH1 and hMSH2 proteins was detected in the 63 sporadic colorectal carcinoma samples by immunohistochemical staining, including tumor tissue, adjacent tissue at 3 cm from the carcinoma, and normal tissue at 10 cm away from the tumor. The positive rate of hMLH1 protein expression in the 63 normal colorectal tissues, adjacent tissues and sporadic colorectal carcinoma tissues was 95.2%, 85.7% and 81.0%, respectively. The positive rate of hMLH1 protein expression was significantly lower in the tumor than in normal colorectal tissues (P < 0.05). The positive rate of hMSH2 protein in the 63 normal colorectal tissues, adjacent tissues and sporadic colorectal carcinoma tissues were 76.2%, 66.7% and 52.4%, respectively. The positive rate of hMSH2 protein expression was significantly lower in the tumor than in normal colorectal tissues (P < 0.01). The positive rate of hMLH1 protein expression was significantly higher in the tumor tissue of patients aged younger than 60 years (100%) than that in patients ≥ 60 years (75.0%, P < 0.05). The positive rate of hMLH1 protein expression in the tumor tissue accompanied by lymphatic metastasis was 50.0%, significantly lower than that (93.3%) in tumors without lymphatic metastasis (P < 0.05). The positive rate of hMSH2 protein expression in the tumor tissue of patients aged younger than 60 years was 80.0%, significantly higher than that (43.8%) in the cases ≥ 60 years (P < 0.05). The positive rate of hMSH2 protein expression in the tumor tissues with invasion reaching to the intestinal serosa (61.5%) was significantly higher than that (37.5%) in the tumors invading to submucosa or muscular layer (P < 0.05). There was a positive correlation between the expressions of hMLH1 and hMSH2 proteins in the sporadic colorectal carcinomas. There is a certain loss of expression of hMLH1 and hMSH2 proteins in sporadic colorectal carcinoma, and is correlated with the age of patients, lymphatic metastasis and different depth of cancer invasion. HMLH1 and hMSH2 may be used as a useful laboratory marker in clinical judgement of occurrence and development of sporadic colorectal carcinoma.

This study aims to investigate the expression levels and values of autophagy genes light chain 3 (LC3) and autophagy-related 5 (ATG5) in intestinal-type gastric cancer. Ninety samples of normal gastric mucosa, intraepithelial neoplasia, and gastric cancer tissue were used in this study. The messenger ribonucleic acid (mRNA) and protein expression levels of autophagy genes LC3 and ATG5 were detected using quantitative reverse transcription polymerase chain reaction, Western blot, and the immunohistochemistry method. The correlations of the autophagy genes and certain clinical pathological parameters were analyzed. The results showed that LC3 mRNA expression was 43.76 ± 20.31 in the normal group, 111.29 ± 18.65 in the intraepithelial neoplasia group, and 131.78 ± 26.29 in the gastric cancer group, while ATG5 mRNA expression was 4.52 ± 2.37 in the normal group, 7.09 ± 1.88 in the intraepithelial neoplasia group, and 10.25 ± 2.81 in the gastric cancer group. The differences between the groups were statistically significant (P < 0.05). The protein expression of LC3 in the normal group was 1.05 ± 0.41, 1.53 ± 0.36 in the intraepithelial neoplasia group, and 1.99 ± 0.14 in the gastric cancer group. The protein expression of ATG5 was 0.78 ± 0.24 in the normal group, 1.37 ± 0.39 in the intraepithelial neoplasia group, and 2.04 ± 0.63 in the gastric cancer group. The differences between the groups were statistically significant (P < 0.05). The positive rate of LC3 protein expression was 33.3% in the normal group and 60% in the intraepithelial neoplasia group, and the difference was statistically significant (χ2 = 4.89; P = 0.04). In the gastric cancer group, the positive rate of LC3 protein expression was 83.3%, making it significantly higher than the intraepithelial neoplasia group, with a statistically significant difference (χ2 = 4.02, P = 0.045). The positive rate of ATG5 protein expression was 23.3% in the normal group, 50.0% in the intraepithelial neoplasia group, and 76.7% in the gastric cancer group. The expression in the intraepithelial neoplasia group was much higher than in the normal group, with a statistically significant difference (χ2 = 4.59, P = 0.03), and that of the gastric cancer group was much higher than that of the intraepithelial neoplasia group, with a statistically significant difference (χ2 = 4.59, P = 0.03). LC3 protein expression was significantly correlated with depth of infiltration, and lymph node status. ATG5 protein expression was significantly correlated with age, depth of infiltration, and lymph node status. There was also a correlation between the LC3 and ATG5 proteins (correlation coefficient r = 0.72, P = 0.001). The enhanced autophagy activity of LC3 and ATG5 may participate in the occurrence and development of intestinal gastric cancer, and they may play a synergistic role in promoting the occurrence and development of intestinal gastric cancer. These findings provide clinical value for the diagnosis of intestinal gastric cancer.

The base excision repair (BER) process removes base damage such as oxidation, alkylation or abasic sites. Two BER sub-pathways have been characterized using in vitro methods, and have been classified according to the length of the repair patch as either 'short-patch' BER (one nucleotide) or 'long-patch' BER (LP-BER; more than one nucleotide). To investigate the occurrence of LP-BER in vivo, we developed an assay using a plasmid containing a single modified base in the transcribed strand of the enhanced green fluorescent protein (EGFP) gene and a stop codon, based on a single-nucleotide mismatch, at varying distances on the 3' side of the lesion. The reversion of the stop codon occurs after DNA repair synthesis and restores EGFP expression after transfection of mismatch-repair-deficient cells. Repair patches longer than one nucleotide were observed for 55-80% or 80-100% of the plasmids with a mean length of 2-6 or 6-12 nucleotides for 8-oxo-7,8-dihydroguanine or a synthetic abasic site, respectively. These data show the existence of LP-BER in vivo, and emphasize the effect of the type of BER substrate lesion on both the yield and the extent of the LP-BER sub-pathway.

Objective To investigate the expressions of 5-lipoxygenase （5-LOX） protein and mRNA in colon mucosa of ulcerative colitis （UC） and to analyze its correlation with endoscopic and histological grading. Methods The biopsies from 32 UC patients and 26 healthy controls were collected, and the expressions of 5-LOX mRNA and protein in colonic mucosa was determined by real-time fluorescent quantitative reverse transcription polymerase chain reaction （RT-PCR） and immunohistochemistry assay, respectively. The endoscopic and histological grading of 32 UC patients were also recorded. Results According to endoscopic grading, there were 10 cases of grade Ⅰ , 19 of grade Ⅱ and 3 of grade Ⅲ in 32 UC patients, and there were 19 cases of grade Ⅰ , 9 of grade Ⅱ and 4 of grade Ⅲ according to histological grading. The expressions of 5-LOX mRNA and protein in UC patients were significantly higher than those in healthy controls （P 〈0. 05 ）. The rate of 5-LOX protein expression increased with endoscopic and histological graObjective To investigate the expressions of 5-lipoxygenase （5-LOX） protein and mRNA in colon mucosa of ulcerative colitis （UC） and to analyze its correlation with endoscopic and histological grading. Methods The biopsies from 32 UC patients and 26 healthy controls were collected, and the expressions of 5-LOX mRNA and protein in colonic mucosa was determined by real-time fluorescent quantitative reverse transcription polymerase chain reaction （RT-PCR） and immunohistochemistry assay, respectively. The endoscopic and histological grading of 32 UC patients were also recorded. Results According to endoscopic grading, there were 10 cases of grade Ⅰ , 19 of grade Ⅱ and 3 of grade Ⅲ in 32 UC patients, and there were 19 cases of grade Ⅰ , 9 of grade Ⅱ and 4 of grade Ⅲ according to histological grading. The expressions of 5-LOX mRNA and protein in UC patients were significantly higher than those in healthy controls （P 〈0. 05 ）. The rate of 5-LOX protein expression increased with endoscopic and histological grades, which was also positively correlated with endoscopic grades （ P 〈 0. 05 ）, but not with histological grades （ P 〉0. 05 ）. Conclusion The expressions of 5-LOX mRNA and protein in colon mucosa from UC patients might be a marker for disease activity. 5-LOX may play a pivotal role in inflammation of UC and be a target site for treatment. Key words: Colitis, ulcerative; 5-lipoxygenase; Reverse transcription polymerase chain reaction; Immunohistochemistry

Hepatocarcinogenesis is a stepwise process during which multiple genes are altered. Understanding the molecular mechanisms that induce hepatocarcinogenesis may improve the screening, prevention and treatment of patients with hepatocellular carcinoma (HCC). In recent years, the oxidored-nitro domain-containing protein 1 (NOR1) gene has been identified to have an important role in the development of HCC in vitro experiments. The current study aimed to examine the expression of NOR1 mRNA and protein expression in specimens of normal liver, hepatitis, cirrhosis and HCC, together representing the process of HCC development. Furthermore, the association between NOR1 expression and clinicopathological parameters of HCC patients was analyzed. Tissue microarrays containing the specimens of human normal liver, hepatitis, cirrhosis and HCC were purchased, and in situ hybridization and immunohistochemistry were used to detect the expression of NOR1 mRNA and protein expression, respectively. It was revealed that the positive rate of NOR1 protein and mRNA expression in the specimens of hepatitis and cirrhosis were not significantly different from that in the normal liver samples. However, the specimens of HCC exhibited an increased positive rate of NOR1 protein and mRNA expression in comparison with the normal liver samples. In addition, a higher positive rate of NOR1 protein expression was observed in HCC patients with a poor pathological differentiation grade and high tumor node metastasis (TNM) stage. In conclusion, the present study provides evidence, for the first time, of the increased expression of NOR1 in human HCC tissues, and its correlation with the pathological stage and TNM status. These findings indicate that NOR1 may be involved in the progression of HCC and it could be employed as a predictive biomarker in HCC development.

Objective To investigate the protein and mRNA expressions of intercellular adhesion molecule-1 (ICAM-1) and monocarboxylate transporters 1 (MCT1) in human gliomas and their correlation. Methods Fifty-eight brain glioma specimens, performed excision and conformed by pathology in our hospital from March 12, 2012 to October 30, 2015, were chosen in our study; in them, 31 were low-grade gliomas (WHO I-II) and 27 were high-grade gliomas (WHO III-IV); another 10 normal brain tissues, collected during the ventriclostomy at the same period were chosen as controls. The mRNA and protein expressions of ICAM-1 and MCT1 were detected by reverse transcription-PCR and two-step immunohistochemical staining (SP method). The relation between ICAM-1 and MCT1 expressions was analyzed by statistical method. Results (1) The ICAM-1 and MCT1 mRNA expressions in normal brain tissues, low-grade tissues and high-grade tissues were increased in sequence, with significant differences (P<0.05); (2) In normal brain tissues, low-grade tissues and high-grade tissues, the positive rate of ICAM-1 protein expression was 2 0.0%, 64.5% and 74.1%, with significant differences (P<0.05); and that of MCT1 protein expression was 30.0%, 58.1% and 70.4%, with significant differences (P<0.05). (3) Positive correlation was noted between ICAM-1 and MCT1 mRNA expressions (r=0.701, P=0.002). Conclusion The higher the glioma malignant degrees, the stronger the protein and mRNA expressions of ICAM-1 and MCT1; they may play important roles together in glioma progress. Key words: Human glioma; Intercellular adhesion molecule-1; Monocarboxylate transporter-1

：Objective To evaluatethe expression of Hedgehog signaling pathway related Sonic hedgehog (SHH) gene incolorectal carcinoma (CRC) and to explore its clinical significance. Methods BetweenDecember 2008 and April 2009, 43 specimens of fresh CRC tissues and adjacent tissues (＞10cm away from the cancer margin) were collected and detected for their expressions of SHHmRNA and proteins with RT-PCR and immunohistochemistry (Envision method), and comparedwith 20 specimens of normal colorectal tissues from non- CRC patients as controls. Theassociations of SHH mRNA and proteins expressions and clinicopathological variables wereanalyzed. Results Gel-imaging analysis displayed a 280 bp band in SHH mRNA-positivespecimens in consistency with theoretical value, which was not seen in SHH mRNA-negativespecimens. The rate of SHH mRNA expression in CRC tissues (27.9%, n=12) was notstatistically different from that in the adjacent tissues (18.6%, n=8) (P＞0.05), but the both were significantly higher than that in the normal mucosa (0%, n=0) (all P＜0.05).The expression intensity of SHH mRNA was significantly higher in CRC tissues than that inthe adjacent tissues (P＜0.05) but not detected in normal colorectal tissues(P＜0.05). Immunohistochemistry demonstrated positive expression of SHHproteins in both CRC tissues and adjacent tissues, appearing as yellow-brown particleswith unstained background in the cell membrane and cytoplasm. The expression of SHHprotein was negative in the cell membrane of normal colorectal mucosa. The rate of SHHprotein expression in CRC tissues was remarkably higher than that in normal colorectalmucosa [25.6% (11/43) vs 0% (0/20) , P＜0.05]. There was nostatistical difference in the positive rate of SHH protein expression between CRC andadjacent tissues, and between adjacent tissues and normal colorectal mucosa (all P＞0.05).The CRC tissues showed the highest intensity of SHH protein expression, followed byadjacent tissues and normal mucosa (all P＜0.05). No significantassociation was found between intensity of SHH mRNA or protein expression in CRC tissuesand the variables such as age, gender, clinical staging, site of tumor, depth of tumorinvasion and pathological classfications (all P＞0.05). Conclusion The SHH mRNA and protein expressions areshown to be significantly enhanced in CRC tissues but not associated with clinical orpathological variables. Key words: Sonic hedgehog; Colorectal neoplasms; Reversetranscriptase polymerase chain reaction; Immunohistochemistry; Clinical significance

Objective To investigate the expression of phosphatase of regenerating liver-3(PRL-3)in Wilms tumor and adjacent kidney tissue and to study the relationship between PRL-3 expression and the clinical stage、pathology grading、recurrence、chemotherapy of Wilms tumor.Methods Reverse transcription polymerase chain reaction(RT-PCR) and Immunohistochemistry S-P were used to measure the expression of PRL-3 in 39 nephroblastoma specimens and 30 controls,which were obtained from adjacent kidney tissues.Itscorrelation with clinical stage、pathological type、recurrence and chemotherapy was analyzed.Results A total of 39 Wilms tumor specimens(30 fresh specimens and 9 paraffin specimens) and 30 fresh adjacent kidney tissues were obtained.3 more recurrent tumor tissues were also obtained.The positive rate of PRL-3 mRNA and protein in Wilms Tumors was 60.0％(18/30) and 61.5％ (24/39)respectively,while that in the adjacent kidney tissues was 13.3％ (4/30)、16.7％(5/30).The expression level was significantly different between Wilms Tumors and adjacent kidney tissues (x2 =5.16、5.34,P＜0.05).The positive expression rate of PRL-3 mRNA and protein in stage Ⅰ、stage Ⅱ 、stageⅢ 、stage Ⅳ of Wilms Tumors was 14.3％(1/7)、57.1％ (4/7)、72.7％(8/11) 、100％ (5/5); 12.5％ (1/8)、50.0％ (4/8)、75.0％ (12/16)、100％ (7/7),respectively.Spearman correlation analysis showed that the expression of PRL-3 mRNA and protein was positively correlated with the clinical stage(r's =0.857、0.839,P＜0.05).The degree of malignancy in tumor tissues also-correlated with the positive expression rate of PRL-3 mRNA.100％ (5/5) expression was seen in tumors with the most severe form,while 52.0％ (13/25) was seen in the favorable type.The trend was similar for protein expression:100％ (5/5) for severe form and 55.9％ (19/34) for favorable form.The expression of PRL-3 mRNA and protein was significantly different between different degree of malignancy tissues (P2tail =0.0049、0.0084＜0.05).Positive expression rate of PRL-3 mRNA and protein in recurrent Wilms tumors were both 100％ (3/3),and their primary tumors all expressed positively.The positive expression rate of PRL-3 mRNA and protein in Wilms tumors trated with pre-operative chemotherapy was 42.9 ％ (3/7) 、45.5 ％ (5/11);while that without pre-operative chemotherapy was 65.2％ (15/23)and 67.9％(19/28).The expression of PRL-3 mRNA and protein was significantly different between pre-operative chemotherapy group and the group without pre-operative chemotherapy (P2tail =0.0348、0.0389＜0.05).Conclusions PRL-3 is widely expressed in Wilms tumor and may be closely related to the clinical stage、degree of malignancy、and recurrence.This would suggest that PRL-3 mRNA and protein may serveas predictors of disease and provide a strong basis for formulating the postoperative treatment. Key words: PRL-3; Nephroblastoma; Reverse transcriptase polymerase chain reaction; Immunohistochemistry

The aim of the present study was to investigate the association between tumor protein 53 (TP53) gene deletion and protein expression and clinical features in esophageal squamous cell carcinoma (ESCC), and to evaluate the predictive value of these two characteristics in the prognosis of ESCC. Immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) were performed to detect the expression of p53 protein and gene deletion in ESCC tissue samples from different ethnic groups in Xinjiang, in order to analyze their association with clinicopathological characteristics and patient prognosis, as well as the sensitivity and specificity of the two methods. In addition, the results were further validated by tissue microarray from a different region. The positive rate of p53 protein expression was 54.5% (201/369) in the multi-ethnic group, and was significantly different between sex (P=0.026) and between tumor differentiation groups (P=0.032). FISH demonstrated that the TP53 gene deletion rate was 31.8% (68/214), which was significantly different between different tumor differentiation (P=0.002), lymph node metastasis (P=0.005) and vascular invasion (P<0.001) groups. The survival rate of patients with TP53 gene deletion was significantly lower than those without TP53 gene deletion (P<0.05). The positive rate of p53 protein expression in the tissue microarray was 58.1% (68/117), which was significantly different between the depth of invasion groups (P=0.011). The TP53 gene deletion rate was 47.9% (56/117), which significantly differed according to lymph node metastasis (P=0.003) and TNM stage (P=0.01). In addition, the total concordance rates of the two methods were 60.3 and 64.1%, respectively. There were also significant differences in the positive rate of TP53 gene deletion and protein expression in different stages of ESCC (P<0.05), which increased gradually with the progression of ESCC. The deletion of the TP53 gene in esophageal cancer was associated with poor prognosis and may be an important biomarker for evaluating the prognosis of patients with ESCC. The combination of FISH and IHC methods could significantly improve the detection rate of TP53 gene abnormalities and the accuracy of prognostic assessment of ESCC.

Ribosome-footprint profiling provides genome-wide snapshots of translation, but technical challenges can confound its analysis. Here, we use improved methods to obtain ribosome-footprint profiles and mRNA abundances that more faithfully reflect geneexpression in Saccharomyces cerevisiae. Our results support proposals that both the beginning of coding regions and codons matching rare tRNAs are more slowly translated. They also indicate that emergent polypeptides with as few as three basic residues within a ten-residue window tend to slow translation. With the improved mRNA measurements, the variation attributable to translational control in exponentially growing yeast was less than previously reported, and most of this variation could be predicted with a simple model that considered mRNA abundance, upstream open reading frames, cap-proximal structure and nucleotide composition, and lengths of the coding and 5' UTRs. Collectively, our results provide a framework for executing and interpreting ribosome-profiling studies and reveal key features of translational control in yeast.

AIMTo determine the association of p53, carcinoembryonic antigen (CEA) and CA19-9 protein expression with esophageal carcinogenesis.METHODSAn iodine staining endoscopic screening program of esophageal lesions was carried out in the high-incidence area of Feicheng County, China. Seventy-seven patients with basal cell hyperplasia (BCH), 247 with low-grade dysplasia (LGD), 51 with high-grade dysplasia (HGD), 134 with invasive cancer, and 80 normal controls diagnosed by mucous membrane biopsy pathology were enrolled. Immunohistochemical detection of p53, CEA and CA19-9 proteins was performed. In the ROC curve analysis, the expression of a single biomarker and the expression of a combination of biomarkers were used to predict the risk of these four esophageal lesions.RESULTSThe positive rates of p53 protein expression in invasive cancer, HGD, LGD, BCH and the normal control groups were 53.0%, 52.9%, 35.6%, 27.3% and 20.0%, respectively; the positive rates of CA19-9 protein expression were 44.0%, 33.3%, 16.5%, 9.2% and 6.2%, respectively; the positive rates of CEA protein expression were 74.6%, 60.8%, 23.3%, 23.7% and 16.2%, respectively. The positive rates of the combined expression of the three biomarkers were 84.3%, 76.5%, 47.6%, 42.9% and 27.5%, respectively. In the receiver operating characteristic curves of the combination of the three biomarkers, the specificity was 88.8% for the normal controls, and the sensitivity was 58.2% for invasive cancer, 25.5% for HGD, 11.2% for LGD, and 6.5% for BCH.CONCLUSIONp53, CEA and CA19-9 protein expression was correlated with esophageal carcinogenesis, and testing for the combination of these biomarkers is useful for identifying high-risk patients with precancerous lesions.

To explore the expressions of SARI (suppressor of AP-1, regulated by IFN) and connective tissue growth factor/cysteine-rich 61/nephroblastoma-1 (CCN1) and clarify their influences on the occurrence, development and prognosis of colorectal carcinoma (CRC). Real-time PCR (polymerase chain reaction) was used to confirm the expressions of SARI and CCN1 at the mRNA level in 32 fresh tissue samples. And the expressions of Caco-2, HT-29 and Lovo were also detected by RT-PCR (reverse transcription-PCR) in cell lines. Tissue specimens were obtained from 116 cases of CRC and the expressions of SARI and CCN1 for each specimen detected by immunohistochemistry. The correlations between the expressions of SARI and CCN1 proteins were summarized. The relationships between the expression levels of SARI and CCN1 and their clinical features in primary CRC were analyzed respectively. The effects of expression levels of SARI and CCN1 proteins on the prognosis were also assessed in 116 CRC cases. The expressions of SARI and CCN1 at the mRNA level in fresh cancerous tissues and cell lines decreased and became up-regulated respectively. The positive rate of SARI protein expression was 76.7% and 28.4% in cancerous and noncancerous tissues respectively (P < 0.05). The positive rate of CCN1 protein expression was 26.7% and 74.1% in cancerous and noncancerous tissues respectively (P < 0.05). A negative correlation was observed between the expressions of SARI and CCN1 (r = -0.24, P < 0.05). The negative expression of SARI correlated with a low grade of differentiation, deep infiltration depth and high TNM staging (P < 0.05). A positive expression of CCN1 correlated with deep infiltration depth and high TNM staging (P < 0.05) while a negative expression of SARI correlated with a lower survival rate than that of a positive expression (χ(2) = 8.47, P < 0.05); additionally, the survival rate of patients with a negative expression of SARI plus a positive expression of CCN1 was further lowered (χ(2) = 12.56, P < 0.05). The aberrant expressions of SARI and CCN1 correlate with the malignant biobehaviors of CRC. And a negative expression of SARI correlates with a worse prognosis of CRC.