Clinical Significance and Prognostic Value of the Expression of LAG-3 and FGL1 in Esophageal Squamous Cell Carcinoma.

In this retrospective study, we analyzed the expression of lymphocyte activating gene 3 (LAG-3) and fibrinogen-like protein 1 (FGP1) mRNA and the corresponding proteins in 78 patients with esophageal squamous cell carcinoma (ESCC) to evaluate the clinical significance and prognostic value. mRNA and protein expression were analyzed by reverse transcription PCR and Western blotting, respectively. The expression of LAG-3 and FGL1 mRNA and the corresponding proteins in tumor tissues were significantly increased in comparison with the normal esophageal mucosa. The overexpression of LAG-3 significantly correlated with the content of tumor-infiltrating lymphocytes (TILs), tumor differentiation, and TNM stage. The overexpression of FGL1 also significantly correlated with TILs, TNM stage, and lymph node metastasis. Kaplan-Meier survival analysis showed that tumor diameter, TNM stage, lymph node metastasis, LAG-3 and FGL1 protein expression were related to the progression-free survival (p<0.05). Multivariate Cox regression showed that the level of FGL1 and TNM stage were independent prognostic factors of progression-free survival. We speculated that the tumor microenvironment of ESCC induces immunosuppression due to up-regulated expression of LAG-3 and FGL1 in the tumor tissues, which promotes tumor growth.

[Application of next-generation sequencing in the pathogenesis of uveal melanoma].

Uveal melanoma, as the most common intraocular malignant tumor in adults, has poor overall survival after metastasis. In recent years, next-generation sequencing technology has been increaingly applied in studying the genetic characteristics of diseases. From the perspectives of genome, epigenome, and transcriptome, this review summed up the genomics mutation, epigenomics regulation mechanism, and immune transcriptomic profiling of uveal melanoma in the context of next-generation sequencing technologies, especially chromosome copy number variation, gene mutation and DNA methylation.

The Awareness and Knowledge Regarding Sarcopenia among Healthcare Professionals: A Scoping Review.

BACKGROUND: Sarcopenia is a prevalent and costly disease associated with serious negative health outcomes, and its prevalence will further grow as the percentage of elderly rises. Healthcare professionals play a crucial role in the prevention, identification and management of sarcopenia and in promoting the well-being of elders. Awareness and knowledge are the prerequisite and basis for these actions. OBJECTIVE: The objective of the review was to summarize available publications to identify the healthcare professionals' awareness and knowledge about sarcopenia, and to identify knowledge gaps that interventions could address. DESIGN: The scoping review will be performed based on the Scoping Review guidelines published by JBI in Australia. METHODS: Six electronic databases, including PubMed, Embase, CINAHL, Web of Science, Cochrane Library and CNKI were searched systematically. Two researchers independently screened the retrieved articles and extracted the information. RESULTS: A total of 6 studies were identified, including 5 quantitative studies and 1 qualitative study. These studies mainly were conducted in Australia, Netherlands and Brazil, and none from Asia. The awareness and knowledge of healthcare professionals about sarcopenia varied in different studies. With exception of one study conducted in oncology clinicians, other studies suggested that awareness and knowledge among healthcare professionals was incomplete and limited. CONCLUSION: The relatively few studies indicated that healthcare professionals had low awareness and limited knowledge of sarcopenia, which could influence and hinder the diagnosis and treatment of sarcopenia in practice. Future researches should develop a rigorously tested and valid sarcopenia knowledge assessment tool and researches conducted in larger samples are needed.

LncRNA LINP1 promotes proliferation and inhibits apoptosis of gastric cancer cells by repressing RBM5.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "LncRNA LINP1 promotes proliferation and inhibits apoptosis of gastric cancer cells by repressing RBM5, by X.-C. Lu, H.-Y. Zhou, J. Wu, Y. Jin, X.-M. Yao, X.-Y. Wu, published in Eur Rev Med Pharmacol Sci 2020; 24 (1): 137-144-DOI: 10.26355/eurrev_202001_19904-PMID: 31957826" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/19904.

Long non-coding RNA AB073614 promotes metastasis of gastric cancer cells by upregulating IGF-2.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Long non-coding RNA AB073614 promotes metastasis of gastric cancer cells by upregulating IGF-2, by X.-Y. Wu, H.-Y. Zhou, X.-M. Yao, X.-D. Chen, J. Wu, X.-C. Lu, published in Eur Rev Med Pharmacol Sci 2020; 24 (1): 145-150-DOI: 10.26355/eurrev_202001_19905-PMID: 31957827" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/19905.

LncRNA LINP1 promotes proliferation and inhibits apoptosis of gastric cancer cells by repressing RBM5.

OBJECTIVE: Recent studies have revealed that long non-coding RNAs (lncRNAs) play a crucial role in tumor progression. Gastric cancer (GC) is one of the common types of malignancies worldwide. This study aimed to identify the exact function of lncRNA LINP1 in the progression of GC. PATIENTS AND METHODS: LINP1 expression in paired cancer tissues and adjacent normal tissues of GC patients was detected by Real-Time quantitative Polymerase Chain Reaction (RT-qPCR). The effect of LINP1 silence on proliferation and apoptosis of GC cells was detected. Meanwhile, the underlying mechanism of LINP1 function was explored by RT-qPCR and Western blot assay. Furthermore, tumor formation assay was performed to examine the ability of LINP1 in tumor formation in vivo. RESULTS: LINP1 expression was remarkably up-regulated in GC tissues compared with adjacent normal tissues. The growth ability of GC cells was significantly inhibited after silencing of LINP1 in vitro. Besides, the apoptosis of GC cells was markedly induced after silencing of LINP1. The silence of LINP1 significantly up-regulated the expression of RBM5 in GC cells. Meanwhile, RBM5 expression in GC tissues was remarkably lower than that of the adjacent normal tissues. Furthermore, tumor formation assay showed that knockdown of LINP1 markedly inhibited tumor formation in vivo. CONCLUSIONS: These results suggested that LINP1 could down-regulate RBM5. Meanwhile, LINP1 remarkably promoted growth ability and suppressed apoptosis of GC in vitro and in vivo. Our findings might provide a novel regulator and therapeutic strategy for GC patients.

Long non-coding RNA AB073614 promotes metastasis of gastric cancer cells by upregulating IGF-2.

OBJECTIVE: Recently, long non-coding RNAs (lncRNAs) have been widely studied for their vital roles in human diseases. In this study, we investigated the effect of lncRNA AB073614 on the metastasis of gastric cancer (GC), and explored the possible underlying mechanism. PATIENTS AND METHODS: AB073614 expression in GC tissue samples was detected by Real-time quantitative polymerase chain reaction (RT-qPCR). The roles of AB073614 in GC metastasis were identified through wound healing assay and transwell assay, respectively. Moreover, RT-qPCR and Western blot assay were used to explore the potential mechanism. RESULTS: AB073614 expression level in GC samples was significantly higher than that of adjacent ones. Besides, the migration and invasion of GC cells were obviously repressed after AB073614 was knocked down. After AB073614 was knocked down in vitro, the mRNA and protein expressions of insulin-like growth factor 2 (IGF-2) was remarkably down-regulated. Furthermore, a negative correlation was found between the expression level of IGF-2 and AB073614 in GC tissues. CONCLUSIONS: AB073614 could promote GC cell migration and invasion via up-regulating IGF-2. Our findings might provide a potential therapeutic target for GC patients.

Mechanism of metformin enhancing the sensitivity of human pancreatic cancer cells to gem-citabine by regulating the PI3K/Akt/mTOR signaling pathway.

OBJECTIVE: To investigate the effect of metformin (MET) on enhancing the sensitivity of human pancreatic cancer cells to gemcitabine (GEM) by regulating the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. MATERIALS AND METHODS: The GEM-resistant human pancreatic cancer PANC-1/GEM cell line was established, and the proliferation ability of PANC-1 and PANC-1/GEM cell lines was detected using the Cell Counting Kit-8 (CCK-8), which was then detected by flow cytometry after they were labeled by Ki67. Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and Western blotting were adopted to detect the difference in the mTOR expression between PANC-1 and PANC-1/GEM cell lines. The proliferation ability of PANC-1/GEM/MET and PANC-1/GEM cell lines was determined using CCK-8 after drug-resistant cell lines were treated with 20 mmol/L MET combined with 0.4 µmol/L GEM or 0.4 µmol/L GEM alone for 48 h. Colony formation assay was applied to detect the proliferation ability of cells. The difference in the expression of mTOR/PI3K/Akt between PANC-1/GEM/MET and PANC-1/GEM cell lines was tested via qRT-PCR and Western blotting, respectively. RESULTS: Compared with PANC-1 cells, PANC-1/GEM cells had significantly enhanced proliferation ability (p<0.01). Flow cytometry results showed that the proliferation ability of PANC-1/GEM cells was notably enhanced (p<0.01). The expression level and phosphorylation level of mTOR in drug-resistant cell lines were increased (p<0.01). After the drug-resistant cell lines were treated with 20 mmol/L MET for 48 h, the proliferation ability of PANC-1/GEM/MET cells was evidently decreased compared with that of PANC-1/GEM cells (p<0.01). The messenger ribonucleic acid (mRNA) and protein expression levels of mTOR/PI3K/Akt were markedly down-regulated (p<0.01). CONCLUSIONS: MET can regulate the PI3K/Akt/mTOR signaling pathway to enhance the sensitivity of human pancreatic cancer cells to GEM.

Therapeutic effect of Bifidobacterium combined with early enteral nutrition in the treatment of severe acute pancreatitis: a pilot study.

OBJECTIVE: To explore the therapeutic effect of Bifidobacterium combined with early enteral nutrition in severe acute pancreatitis. PATIENTS AND METHODS: A total of 60 patients with severe acute pancreatitis admitted from November 2012 to November 2016 were retrospectively analyzed. According to the different treatment methods, the patients were divided into Bifidobacterium combined with early enteral nutrition group (experiment group) and early enteral nutrition group (control group). Serum ALB (albumin), CRP (C-reactive protein), WBC (white blood cell count) and PCT (procalcitonin) levels in both groups were observed. The pain relief time, diet recovery time, length of stay, and hospitalization costs between the two groups, were compared. The APACHE (acute physiology and chronic health evaluation scoring system) II score and SOFA (sequential organ failure assessment) score before and after nutritional support were compared between the two groups. Adverse events and complications were observed as well. RESULTS: 58 patients recovered and 2 died after treatment. Improvements in laboratory indicators such as ALB, CRP, WBC and PCT were much better in the experiment group than the control group (p<0.05). Both the length of days and hospitalization cost were lower in the experiment group than those of the control group (p=0.0029, p=0.0435). In the comparison of hospitalization symptoms, shorter pain relief time and diet recovery time were found in the experiment group than those in the control group (p=0.0003, p=0.0218). After the treatment, APACHE II score and SOFA score of the experiment group were also higher than the control group. No significant differences in adverse events and complications between the two groups were exerted (p>0.05). CONCLUSIONS: Bifidobacterium combined with early enteral nutrition can improve the nutritional status of patients with severe acute pancreatitis in the acute stage, which also enhances the patient's immune capacity and the body's resistance to disease.

Expression and clinical significance of long-non-coding RNA GHET1 in pancreatic cancer.

OBJECTIVE: To investigate the expression level and biological function of long non-coding RNA gastric carcinoma high expressed transcript 1 (lncRNA-GHET1) in pancreatic ductal adenocarcinoma (pancreatic cancer for short), to analyze the correlation between the expression of GHET1 and clinicopathological features and to explore the role and clinical significance of GHET1 in the development and progression of pancreatic cancer. PATIENTS AND METHODS: The relative expression of GHET1 in 5 human pancreatic cancer cell lines was detected by quantitative Real-time polymerase chain reaction (qRT-PCR). The specific interference sequence of GHET1 was designed and transiently transfected into pancreatic cancer cells. qRT-PCR assay was used to detect the interference efficiency. Cell counting kit-8 (CCK-8) assay was used to detect the effect of the interference with GHET1 on the proliferation of pancreatic cancer cells. Flow cytometry was used to detect the effect of the interference with GHET1 on the cycle distribution and apoptosis. qRT-PCR was used to detect the relative expression of GHET1 in pancreatic cancer tissues compared with that in cancer-adjacent tissues. The correlation between the expression of GHET1 and the pathological features of pancreatic cancer patients was analyzed. RESULTS: The expression of GHET1 in human pancreatic cancer cells was relatively high. The results of CCK-8 showed that the proliferation of tumor cells was inhibited after the interference with GHET1 expression. The results of flow cytometry showed that the expression of GHET1 was blocked at G1/G0 phase, and the apoptosis rate was increased. The results of qRT-PCR showed that the expression of GHET1 was upregulated in pancreatic cancer tissues of 49 out of 64 patients compared with that in cancer-adjacent tissues, and the highly expressed GHET1 was positively correlated with the tumor, node and metastasis (TNM) staging of pancreatic cancer. CONCLUSIONS: Highly expressed GHET1 promotes the proliferation of pancreatic cancer, inhibits apoptosis and is related to TNM staging. The expression of GHET1 can be used as a potential molecular marker for the prognosis and therapeutic target of pancreatic cancer.

Metformin regulates tight junction of intestinal epithelial cells via MLCK-MLC signaling pathway.

OBJECTIVE: To observe the effect of metformin on the tight junction of intestinal epithelial cells and its relevant mechanism. MATERIALS AND METHODS: Caco-2 cell monolayers were incubated with or without tumor necrosis factor-α (TNF-α) (10 ng/mL) in the absence or presence of indicated concentrations of metformin. Transepithelial electrical resistance (TEER) was measured at various time points. Caco-2 cell permeability was assessed using fluorescein permeability test. Immunofluorescence was used to detect the distribution of tight junction protein. Western blotting and Real-Time PCR were used to detect the expression of tight junction protein and Myosin light chain kinase (MLCK)-Myosin light chain (MLC) signaling pathway. RESULTS: Metformin attenuates the effects of TNF-α on Caco-2 cell TEER and paracellular permeability, prevents TNF-α-induced morphological disruption of tight junctions, ameliorates the inhibiting effect of TNF-α on epithelial tight junction-related protein expression and suppresses the TNF-α-induced increase in MLCK production. CONCLUSIONS: Metformin can stabilize and up-regulate tight junction protein by inhibiting MLCK-MLC signaling pathway, thus ameliorating the tight junction of intestinal epithelial cells.

High expression of LncRNA CASC15 is a risk factor for gastric cancer prognosis and promote the proliferation of gastric cancer.

OBJECTIVE: To investigate the expression of long non-coding RNA CASC15 in gastric cancer tissue and its effect on the proliferation of gastric cancer cell line MKN28. MATERIALS AND METHODS: We found that expression of lncRNA CASC15 in gastric cancer tissue was higher than normal gastric epithelium through the TCGA and Gene Expression Omnibus (GEO) database. Then, we detect the RNA level of CASC15 from clinical samples of 42 normal gastric epithelial tissues and 60 gastric cancer tissues. In order to explore the function of CASC15 in gastric cancer, we perform gain-function and loss-function assay in gastric cancer cell lines. RESULTS: We found that expression of lncRNA CASC15 in gastric cancer tissue was higher than normal gastric epithelium through the TCGA database and the related microarray data set was searched from Gene Expression Omnibus (GEO) database.  Then, we extracted total RNA from clinical samples of 42 normal gastric epithelial tissues and 60 gastric cancer tissues. The results of quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) were consistent with those of TCGA analysis. Clinical data analysis showed that the expression of LncRNA CASC15 was correlated with the total survival, tumor size and TMN staging in clinical patients. Clinical data analysis showed that the expression level of CASC15 was correlated with tumor size and TNM stage in clinical patients. Compared with the negative control group, the proliferation and cell cloning ability of MKN28 cells overexpressing LncRNA CASC15 significantly increased (p<0.001), indicating that overexpression of LncRNA CASC15 promoted the proliferation of MKN28 cells. CONCLUSIONS: The expression of LncRNA CASC15 was significantly higher in gastric cancer tissues and its expression was negatively correlated with the overall survival of clinical patients. It was positively correlated with the tumor size and TMN stage. LncRNA CASC15 could promote the proliferation of gastric cancer cells and was expected to become the molecular marker for prediction and prognosis of gastric cancer, as well as a potential therapeutic target.

[Efficacy and safety analysis of different dose idarubicin plus cytarabine regimen as induction chemotherapy for young patients with de-novo acute myeloid leukemia].

OBJECTIVE: To compare the efficacy, safety and long-term prognosis between different dose idarubicin (IDA) combined with cytarabine (IA) as induction chemotherapy in newly diagnosed young patients of acute myeloid leukemia (AML). METHODS: A total of 149 newly diagnosed young AML patients (APL excluded) between January 2009 to July 2014 was enrolled. According to the dose of IDA, the patients were divided into three groups, high standard- dose IA group (10- 12 mg · m (- 2) · d(- 1)), low standard-dose IA group (8-9 mg·m(-2)·d(-1)) and low-dose IA group (<8 mg·m(-2)·d(-1)). The efficacy, adverse effects and long- term prognosis among the three groups were compared. RESULTS: Of them, 34 patients were in high standard-dose IA group, 53 in low standard-dose IA group and 62 in low-dose IA group. After one cycle of induction chemotherapy, the complete remission (CR) rate was 79.4%, 75.5% and 46.8%, the overall response (OR) rate was 97.1%, 94.3% and 64.5%, and the overall CR rate was 85.3%, 81.1% and 54.8%, respectively. Compared with low- dose IA group, high standard- dose IA group and low standard-dose IA group had significantly better result (P<0.05), but there was no significant difference between the latter two groups (P>0.05). Multivariate analysis also showed that standard-dose IA was favorable factor for induction chemotherapy (P<0.05). The adverse effects were similar in the three group, other than the lowest count of WBC (P=0.002). Low standard-dose IA can improve the OS compared to the low-dose IA (P=0.003), but EFS, RFS was similar in the three groups. CONCLUSIONS: For the newly diagnosed young(<55) AML patients, the standard-dose IA has better CR rate. The adverse effects were similar in the three groups. High-dose IA may improve the OS compared to the low-dose IA.

Interleukin-10 polymorphisms associated with susceptibility to acute myeloid leukemia.

We investigated the association between polymorphisms in interleukin-10 (IL-10) -1082G/A (rs1800896), -819T/C (rs1800871), and -592A/C (rs1800872) and the risk of acute myeloid leukemia (AML) in a Chinese population. A total of 167 primary AML cases and 328 healthy control subjects were recruited at the First People's Hospital of Yunnan Province between March 2009 and January 2012. The polymorphisms rs1800896, rs1800871, and rs1800872 were genotyped by polymerase chain reaction-restriction fragment length polymorphism. Multivariate regression analyses showed that subjects carrying the rs1800871 CC genotype and C allele had a significantly increased risk of AML, with adjusted odds ratios (95% confidence intervals) of 1.72 (1.01-2.97) and 1.38 (1.04-1.81), respectively. Those carrying the rs1800872 G allele had a slightly increased risk of AML, with an adjusted odds ratio (95% confidence interval) of 1.30 (1.01-1.72). Moreover, genotyping results demonstrated that subjects carrying both the rs1800871 C allele and rs1800872 G allele had a moderately increased risk of AML, indicating that the 2 genotypes had a synergistic effect on AML risk (odds ratio = 2.03, 95% confidence interval = 1.24- 3.15). Our results demonstrated that polymorphisms in rs1800871 and rs1800872 enhance the risk of AML, and these 2 single nucleotide polymorphisms have a synergistic effect on AML risk.

Antitumor and antimetastatic activities of grape skin polyphenols in a murine model of breast cancer.

Treatment modalities are not effective once breast cancer metastasis has occurred. Dietary botanicals may have a better protective effect. We therefore investigated the effects of grape skin polyphenols on a highly metastatic mouse mammary carcinoma cell line. In vitro treatment of 4T1 cells, with grape skin polyphenols resulted in inhibition of the migration and viability in a dose-dependent manner. The migration of 4T1 cells was significantly inhibited by grape skin polyphenols, even at a very low concentration (5 µg/ml), and was totally inhibited when the concentration was 20 µg/ml. However, 20 µg/ml of grape skin polyphenols inhibited cell viability by only 11.4%. The inhibition of migration is independent of decreased cell viability or apoptosis induction. Further analysis indicated that the inhibition of migration by grape skin polyphenols is involved in blocking the PI3k/Akt and MAPK pathways. The effects of dietary grape skin polyphenols were then examined using an in vivo model in which 4T1 cells were implanted subcutaneously in Balb/c mice. The metastasis of tumor cells to the lungs was inhibited significantly by dietary grape skin extracts (0.5 and 1.0 mg/ml in drinking water) and the survival of the mice enhanced. These data suggest that grape skin polyphenols possess chemotherapeutic efficacy against breast cancer with metastases.

Simvastatin protects diabetic rats against kidney injury through the suppression of renal matrix metalloproteinase-9 expression.

OBJECTIVE: To observe the effects of simvastatin on urinary excretion of matrix metalloproteinase-9 (MMP- 9), renal expression of MMP-9, and investigate its possible renoprotective mechanisms in streptozotocin (STZ)-induced diabetic rats. METHOD: Twenty-four Wistar rats were divided into 3 groups: control healthy rats (group C, no.=8), untreated diabetic rats (group D, no.=8), and diabetic rats treated with simvastatin (20 mg/kg/d) (group S, no.=8). Peripheral blood glucose was tested weekly, glycosylated hemoglobin A1c (HbA1c), total cholesterol (TC), LDL cholesterol (LDL-C) levels, and urinary albumin (ALB) excretion rate as well as the urinary excretion rates of retinol-binding protein (RBP) and MMP-9 were tested at 8th week. The renal tissues of diabetic rats were obtained for evaluating kidney/ body weight ratio, observing renal pathological changes by electron microscope and examining the expression of renal MMP-9 mRNA by RT-PCR. RESULTS: There was no statistical difference on the change of peripheral blood TC and LDL-C between group C and group D. Peripheral blood glucose, HbA1c levels kidney/body weight ratio urinary excretion rates of ALB, RBP, and MMP-9 concurrently with the expression of renal MMP-9 mRNA were significantly higher in groups D and S compared with group C (p<0.01). Treatment with simvastatin significantly lowered peripheral blood TC, LDL-C, kidney/body weight ratio, urinary excretion rates of ALB, RBP, and MMP-9 as well as the expression of renal MMP-9 mRNA (p<0.01); however, there was no evident effect on the change of blood glucose and HbA1c levels between group D and group S. In addition, urinary excretion rate of MMP-9 showed positive correlations with the urinary ALB excretion and urinary RBP excretion. Pathological lesions of the glomeruli and epithelial cells foot processes (FP) was lightened by simvastatin. CONCLUSION: Simvastatin may has a potential therapeutic target in diabetic nephropathy, which may be partly attributed to down-regulating over-expression of MMP-9 in renal tissue.

Intensive insulin therapy decreases urinary MCP-1 and ICAM-1 excretions in incipient diabetic nephropathy.

BACKGROUND: Nowadays, intensive insulin treatment has been widely used in type 2 diabetics who have poor control of blood glucose, to reduce the risk of chronic complications of diabetes. Recently, some scholars have paid more attention to the pivotal role of inflammation involved in type 2 diabetes and its complications. Monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1), which are two important inflammatory chemokines, have been documented to participate in the onset and development of type 2 diabetes and its complications, such as diabetic nephropathy (DN). DESIGN: In the current study, we recruited 30 type 2 diabetics with microalbuminuria to be treated with multiple insulin injections daily for 2 weeks. Random spot urine samples (corrected for creatinine-Cr) were collected for the examination of urinary MCP-1, ICAM-1 and albumin (Alb) levels before and after the intensive insulin therapy. Changes in their levels were observed to test the hypothesis that type 2 diabetes with microalbuminuria is associated with elevated urinary concentrations of MCP-1 and ICAM-1, and intensive insulin therapy can result in a decline of Alb by reducing the inflammatory reaction. RESULTS: The urinary MCP-1/Cr and urinary ICAM-1/Cr ratios in type 2 diabetic patients with microalbuminuria were much higher than those in normal controls, and intensive insulin treatment could decrease significantly the urinary MCP-1/Cr, ICAM-1/Cr and Alb/Cr ratios in type 2 diabetics with microalbuminuria. CONCLUSION: Intensive insulin treatment may protect against renal injury in early DN by reducing the urinary MCP-1 and ICAM-1 excretions.

Symmetric and asymmetric DNA methylation in the human IGF2-H19 imprinted region.

The two contiguous IGF2 (human insulin-like growth factor II) and H19 genes are reciprocally imprinted in both human and mouse. In most tissues, IGF2 is transcribed only from the paternal chromosome while H19 is transcribed only from the maternal allele. The presence of a differential methylation region (DMR) on the two parental alleles at the 5' flanking region of H19 has been proposed to constitute the gametic imprint, which controls the reciprocal allelic expression of the two genes. Using bisulfite genomic sequencing, we have assessed the methylation status of cytosine (including 154 CpG sites) in six CpG-rich regions of the human IGF2-H19 genes. In a CpG island near promoter P3 of the IGF2 gene, more than 99.8% of all cytosines were converted to thymidine by sodium bisulfite mutagenesis, indicating that none of the CpGs was methylated. In the IGF2 exon 8-9 region, mosaic methylation of 56 CpG sites was observed in fetal tissues and in adult blood DNA. In contrast to the mosaic methylation of IGF2, the allelic methylation of the human H19 DMR was uniform. In the CpG region located 2 kb upstream (-2362 to -1911) of the H19 transcription site, all 25 CpG sites were completely methylated on only one parental allele. Uniform allele-specific methylation was also observed in the CpG island proximal to the H19 promoter (-711 to -290) with complete methylation of all 25 CpG sites in one parental allele. In contrast, the CpG region in the H19 promoter (-292 to +15) was mosaically methylated in all tissues. In addition, cytosine was methylated at three CpNpG and GpNpC sites on the top DNA strand and one CpNpG site on the bottom DNA strand from the fetal brain. The cytosines at CpG sites were methylated on both DNA strands (symmetric methylation) while cytosines at the CpNpG and GpNpC sites were methylated on only one DNA strand (asymmetric methylation). The asymmetric methylation was associated with tissue-specific disruption of H19 genomic imprinting in fetal brain.