Antiporters ; genetics ; Base Sequence ; Female ; Glycogen Storage Disease Type I ; diagnosis ; genetics ; Humans ; Infant ; Monosaccharide Transport Proteins ; genetics ; Mutation ; Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo assess the relationship between the overexpression of facilitative glucose transporter-1 (Glut1) and fluorine-18 fluorodeoxyglucose (FDG) uptake in patients with primary lung squamous cell carcinoma.

METHODSFrom April 1999 to March 2001, 23 patients with lung squamous cell carcinoma were imaged using FDG positron emission tomography (PET) before surgery. Their maximum and mean standard uptake values (SUVmax and SUVmean) of tumor and SUV of the normal lung (SUVlung) were measured. The expression of Glut1 of all the 23 cases was analysed in paraffin sections using SP immunohistochemistry.

RESULTSAll the 23 tumors tested were Glut1 positive (69 +/- 18)% of tumor cell area was positive and staining intensity was 4.6 +/- 0.7. All tumors of the patients could be detected by FDG-PET. FDG uptake of tumor was higher than that of normal lung (P < 0.01). SUVmax, SUVmean and SUVlung were 8.33 +/- 4.14, 6.10 +/- 3.00 and 0.38 +/- 0.13 respectively. Correlations were found among Glut1 expression and FDG uptake and tumor size (P < 0.01).

CONCLUSIONS(1) Glut1 overexpression is universal in the lung squamous cell carcinoma. (2) SUV was higher in the lung squamous cell carcinoma than that of the normal lung tissue. (3) Glut1 expression and FDG uptake and tumor size appear to be correlated with each other in patients with lung squamous cell carcinoma.

Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; Fluorodeoxyglucose F18 ; metabolism ; Glucose Transporter Type 1 ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; Male ; Middle Aged ; Monosaccharide Transport Proteins ; analysis

OBJECTIVEGlycogen storage disease type Ib (GSDIb) is caused by a deficiency of glucose-6-phosphate translocase (G6PT) activity due to SLC37A4 gene mutations. Most GSDIb patients have recurrent infections and inflammatory bowel disease, with poor prognosis. Detection of SLC37A4 gene mutations is of great significance for the diagnosis, subtyping and outcome prediction of GSD patients. This study aims to analyze SLC37A4 gene mutations in Chinese GSDIb patients and to investigate the relationship between its genotypes and clinical manifestations.

METHODSAll exons and their flanking introns of SLC37A4 gene in 28 Chinese children with a primary diagnosis of GSDIb were screened by PCR combined with direct DNA sequencing to detect SLC37A4 gene mutations.

RESULTSFive SLC37A4 gene mutations were detected in 7 (25%) of the 28 children, i.e., p.Gly149Glu (9/13, 69%), p.Gly115Arg (1/13, 8%), p.Pro191Leu (1/13, 8%), c.959-960 insT (1/13, 8%) and c.870+5G>A (1/13, 8%).

CONCLUSIONSIn this study, c.959-960 insT is a novel mutation and p.Gly149Glu is the most common mutation. p.Gly149Glu may be associated with severe infections in children with GSDIb.

Antiporters ; genetics ; Child, Preschool ; Female ; Glycogen Storage Disease Type I ; complications ; genetics ; Humans ; Infant ; Male ; Monosaccharide Transport Proteins ; genetics ; Mutation ; Sequence Analysis, DNA

The mechanism of glucose transported (GT) expression on the plasma membranes of hepatoma cells in rats induced by 3'-methyl-4-dimethylaminoazobenzene (3'-MeDAB) was studied. Cytochalasin B binding to plasma membrane fractions from control and 3'-MeDAB group in the absence of cold cytochalasin B showed 9,825 +/- 925 and 30,165 +/- 625 dpm/mg membrane protein. Scatchard plot analysis showed that the GTs present on the plasma membrane fractions in control and 3'-Me DAB groups were 5.0 and 16.0 pmol/mg membrane protein and their Kd values were 151 and 157 nM, respectively. These results suggest that the numbers of GTs in plasma membrane were increased in the 3'-Me DAB group compared to the control group. In contrast, the amounts of GTs in low density microsomal (LDM) fractions measured by a photoaffinity labeling technique using [3H]-cytochalasin B were 31,207 and 11,702 dpm/mg protein in the control and 3'-Me DAB group, respectively. These results suggest that GTs were translocated from LDM to plasma membranes during carcinogenesis. To confirm these results by an independent method 10% SDS-polyacrylamide gel electrophoresis was carried out. Gel slice No. 13 corresponding to MW of 45 kDa from plasma membrane fractions showed increased radioactivities in the 3'-Me DAB group compared to the control group. However, LDM fractions of the 3'-Me DAB group showed decreased radioactivities compared to the control group. Western blot analysis using anti-human RBC GT antibody present in the plasma membranes and LDM fractions from control and 3'-Me DAB groups did not show any significant difference, indicating low cross-reactivity between them. These results indicate that increased glucose transport seems to be more likely due to reciprocal redistribution of GTs between plasma membrane and LDM fractions.
Animal ; Blotting, Western ; Cell Membrane/chemistry ; Cytochalasin B/metabolism ; Glucose/*analysis ; Liver Neoplasms, Experimental/*metabolism ; Male ; Methyldimethylaminoazobenzene ; Microsomes, Liver/*chemistry ; Monosaccharide Transport Proteins/*analysis ; Rats ; Support, Non-U.S. Gov't

OBJECTIVEGlycogen storage disease type Ib (GSDIb, MIM: 232220) is an autosomal recessive inborn error of metabolism caused by deficiency of the glucose-6-phosphate translocase. The clinical manifestations include symptoms and signs of both the typical GSDIa, including hepatomegaly, fasting hypoglycemia, lactic acidemia and hyperlipidemia, and the dysfunction of neutrophils of recurrent infection and neutropenia. More than 84 mutations have been identified since the discovery of the SLC37A4 gene as the disease causing gene. Up to date, 5 mutations in 4 Chinese patients were reported from Hong Kang and Taiwan. In order to see the spectrum of the SLC37A4 gene mutations and the correlation between genotype and phenotype in patients with GSDIb of the mainland of China, the authors investigated 17 GSDIb patients from 15 families in this study.

METHODData of 17 patients from 12 provinces, 11 male and 6 female, aged 6 months to 35 years, were collected from the genetic clinics of Peking Union Medical College Hospital from Oct. 2006 to Mar. 2009. All of them were Han Chinese in ethnicity. Consanguineous status was confirmed in 2 unrelated patients. All patients were presented with hepatomegaly, fasting hypoglycemia, lactic acidemia, hyperlipidemia and neutropenia with variable frequency of infections. The full coding exons, their relevant exon-intron boundaries, and the 5'- and 3'-flanking regions of the SLC37A4 gene were amplified and directly sequenced. RT-PCR was performed to verify the effect of the 2 novel splicing mutations.

RESULTA total of 11 mutations were identified in 15 families. Four mutations, p.Gly149Glu, p.Pro191Leu, p.Arg415X and c.1042_1043 del CT, were previously reported, and seven mutations, p. Leu23Arg, p.Gly115Arg, p.Gly281Val, p.Arg415Gly, c.784 + 1G > A, c.870 + 5G > A and c.1014_1120del107, were novel. The frequent mutations are p.Pro191Leu, p.Gly149Glu and c.870 + 5G > A, accounting for 37%, 15% and 11% of mutant alleles respectively. RT-PCR analysis of novel mutation c.784 + 1G > A confirmed the splicing of exon 5 of 159 bp, causing inframe deletion. While mutation c.870 + 5G > A was proved to cause exon 6, 86 bp, deletion causing frame-shift. Among 15 families, 12 genotypes were identified, including 3 with homozygous mutation and 9 with compound heterozygous mutations. Homozygous p.Pro191Leu mutation was the only genotype detected in more than 1 family and was found in 4 unrelated families, including 1 patient from consanguineous marriage.

CONCLUSIONA total of 11 SLC37A4 gene mutations were identified in 15 families of the mainland of China. The frequent mutations are p.Pro191Leu, p.Gly149Glu and c.870 + 5G > A. The number of Chinese SLC37A4 gene mutations was extended from 5 to 14.

Adolescent ; Adult ; Antiporters ; genetics ; Child ; Child, Preschool ; DNA Mutational Analysis ; Female ; Genotype ; Glycogen Storage Disease Type I ; genetics ; Humans ; Infant ; Male ; Monosaccharide Transport Proteins ; genetics ; Mutation ; Pedigree ; Young Adult

Increased expression of glucose transporter1 (GLUT1) has been reported in many human cancers. We hypothesized that the degree of GLUT1 might provide a useful biological information in gastric adenocarcinoma. RT-PCR and immunostaining were used to analyze GLUT1 expression in gastric cancer. RT-PCR showed GLUT1 expression was not largely detected in normal gastric tissue but was detected in cancerous gastric tissue of counterpart. By immunohistochemistry, GLUT1 protein was absent in normal gastric epithelium and intestinal metaplasia. 11 of 65 patients with gastric adenocarcinoma had specific GLUT1 immunostaining in a plasma membrane pattern with varied intensities. GLUT1 protein did not show any significant correlation with tumor stage and nodal metastasis (p+AD4-0.05 by Mann-Whitney test). However, the positive immunostaining for GLUT1 is associated with intestinal differentiation (p+AD0-0.003). Our results suggest that GLUT1 protein is associated with intestinal type of gastric cancer.
Adenocarcinoma/pathology ; Adenocarcinoma/chemistry+ACo- ; Adult ; Aged ; Female ; Gastric Mucosa/pathology ; Gastric Mucosa/chemistry+ACo- ; Human ; Intestines ; Male ; Metaplasia ; Middle Age ; Monosaccharide Transport Proteins/analysis+ACo- ; Neoplasm Proteins/analysis+ACo- ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms/pathology ; Stomach Neoplasms/chemistry+ACo- ; Tumor Markers, Biological/analysis+ACo-

Increased expression of glucose transporter1 (GLUT1) has been reported in many human cancers. We hypothesized that the degree of GLUT1 might provide a useful biological information in gastric adenocarcinoma. RT-PCR and immunostaining were used to analyze GLUT1 expression in gastric cancer. RT-PCR showed GLUT1 expression was not largely detected in normal gastric tissue but was detected in cancerous gastric tissue of counterpart. By immunohistochemistry, GLUT1 protein was absent in normal gastric epithelium and intestinal metaplasia. 11 of 65 patients with gastric adenocarcinoma had specific GLUT1 immunostaining in a plasma membrane pattern with varied intensities. GLUT1 protein did not show any significant correlation with tumor stage and nodal metastasis (p+AD4-0.05 by Mann-Whitney test). However, the positive immunostaining for GLUT1 is associated with intestinal differentiation (p+AD0-0.003). Our results suggest that GLUT1 protein is associated with intestinal type of gastric cancer.
Adenocarcinoma/pathology ; Adenocarcinoma/chemistry+ACo- ; Adult ; Aged ; Female ; Gastric Mucosa/pathology ; Gastric Mucosa/chemistry+ACo- ; Human ; Intestines ; Male ; Metaplasia ; Middle Age ; Monosaccharide Transport Proteins/analysis+ACo- ; Neoplasm Proteins/analysis+ACo- ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms/pathology ; Stomach Neoplasms/chemistry+ACo- ; Tumor Markers, Biological/analysis+ACo-

OBJECTIVETo analyze the clinical and SLC2A1 gene mutation characteristics of glucose transporter type 1 deficiency syndrome.

METHODThe detailed clinical manifestations of six cases were recorded. The laboratory tests including EEG, MRI, blood chemistry, and lumbar puncture were performed. SLC2A1 gene mutations were analyzed by PCR, DNA sequencing and multiplex ligation-dependent probe amplification (MLPA).

RESULTPatient 1, 2 and 3 had classical clinical symptoms including infantile onset seizures, development delay. Patient 4, 5 and 6 had non-classical clinical symptoms including paroxysmal behavior disturbance, weakness, ataxia, lethargy, especially after fasting or exercise, without severe seizures. The plasma glucose levels were normal. The CSF glucose levels decreased in all the six cases, ranged from 1.10 mmol/L to 2.45 mmol/L, the mean level was 1.68 mmol/L. The CSF glucose/plasma glucose ratios decreased, ranged from 0.16 to 0.51, the mean ratio was 0.34. Four patients had normal EEG. Two patients had focal and diffuse epileptiform discharge, and one of them also had paroxysmal occipital or generalized high-amplitude slow waves during awake and sleep time. MRI abnormalities were found in three patients, patient 1 with mild brain atrophy, patient 3 with bilateral ventricle plump, and patient 4 with high signals in T2 in the frontal and occipital white matter, interpreted as hypomyelination. SLC2A1 gene mutations were found in six cases. Patient 1 has large scale deletion in exon 2. In patient 2 to 6, the mutations were c.741 G>A (E247K), 599delA, 761delA, c.1148 C>A (P383H), c.1198 C>T (R400C) respectively. Two patients were treated with ketogenic diet. The seizures disappeared and development became normal. Three patients responded to frequent meals with snacks. One patient refused any treatments, the symptoms continued to exist.

CONCLUSIONThe clinical manifestations of glucose transporter type 1 deficiency syndrome are varied. The common symptoms included infantile onset seizures and various paroxysmal events. These neurologic symptoms generally fluctuated and were influenced by factors such as fasting or fatigue. This feature could be a very important clue for the diagnosis of GLUT1-DS. Lumbar puncture is recommended in patients with episodic CNS symptoms especially after fasting. GLUT1-DS is a treatable neurometabolic disorder, early diagnosis and treatment may improve the prognosis of the patients.

Biomarkers ; analysis ; Brain ; diagnostic imaging ; pathology ; Carbohydrate Metabolism, Inborn Errors ; diagnosis ; genetics ; therapy ; Child ; Child, Preschool ; DNA Mutational Analysis ; Diet, Ketogenic ; Electroencephalography ; Epilepsy ; diagnosis ; genetics ; therapy ; Female ; Follow-Up Studies ; Glucose Transporter Type 1 ; genetics ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Monosaccharide Transport Proteins ; deficiency ; genetics ; Movement Disorders ; diagnosis ; genetics ; therapy ; Mutation ; genetics ; Radiography