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Resumen La deficiencia de fructosa-1,6-bisfosfatasa (deficiencia de FBPasa) es un defecto metabólico congénito poco común que afecta la gluconeogénesis. Es una enfermedad genética autosómica recesiva. El paciente se presenta con hipoglucemia en ayunas y acidosis metabólica, y puede tener hiperventilación, apnea y cetosis. Aunque la enfermedad puede ser fatal en el período neonatal, el tratamiento adecuado puede producir un pronóstico excelente. A continuación, presentamos una paciente de 21 años con déficit de fructosa-1,6-bisfosfatasa, quien presentó cuadro gastroenteritis viral que provocó descompensación de su patológica de base, la paciente presentó evolución satisfactoria al manejo con cristaloides y dextrosa endovenosa. Se expone este caso porque es una entidad de baja frecuencia, con escasos reportes en adultos y con adecuada respuesta al tratamiento dietario.
Abstract Fructose-1,6-bisphosphatase deficiency (FBPase deficiency) is a rare congenital metabolic defect affecting gluconeogenesis. It is an autosomal recessive genetic disease. The patient presents with fasting hypoglycemia and metabolic acidosis, and may have hyperventilation, apnea, hypoglycemia, and ketosis. Although the disease can be fatal in the neonatal period, appropriate treatment can produce an excellent prognosis. Here we present the case of a 21-year-old patient with fructose-1,6-bisphosphatase deficiency, who presented with viral gastroenteritis that caused decompensation of her underlying pathology, the patient presented satisfactory evolution with crystalloids and intravenous dextrose. This case is presented because of its low frequency, with few reports in adults and with adequate response to dietary treatment.
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Objective:To explore the effects of enriched environment on neurological function in cerebral ischemia-reperfusion injury rats and the glucose metabolism in ischemic penumbra. Methods:A total of 72 adult male Sprague-Dawley rats were randomly divided into sham group (n = 24), model group (n = 24) and enriched environment group (n = 24). The latter two groups suffered cerebral ischemia 60 minutes and reperfused with modified Longa's method. The enriched environment group was fed in enriched environment after operation. All the rats were assessed with modified Neurological Severity Score (mNSS) before, and one, seven, 14, 21 and 28 days after operation. One and 28 days after operation, twelve rats from each groups were sacrificed after mNSS assessment, respectively. The histopathology was observed with HE staining. The expressions of hypoxia-inducible factor-1α (HIF-1α), glucose transporter 1 (GLUT1) and 6-phosphofructo-2-kinase/fructose-2-bisphosphatase 3 (PFKFB3) in ischemic penumbra were determined with reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting. The levels of ATP, ADP and AMP in ischemic penumbra were measured with high performance liquid chromatography (HPLC) and the energy charge (EC) was calculated. Results:Compared with the model group, the scores of mNSS decreased in the enriched environment group since 14 days after operation (P < 0.05). The cells in the penumbra presented edema, nuclear pyknosis marginalization, vacuolar arrangement and other pathological changes in the model group and the riched environment group one day after operation; while compared with the model group, the levels of ATP and EC decreased and the mRNAs and protein expression of HIF-1α, GLUT1 and PFKFB3 increased in the enriched environment group (P < 0.05). The pathology improved in the riched environment group compared with that in the model group 28 days after operation; while the mRNAs and protein expression of HIF-1α, GLUT1 and PFKFB3 increased, as well as the levels of ATP and EC (P < 0.05). Conclusion:Enriched environment can promote the recovery of neurological function in rats after cerebral ischemia-reperfusion, which may associate with promoting expression of HIF-1α and downstream GLUT1 and PFKFB3, and improving glucose metabolism.
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Objective@#To construct the recombinant adenoviral containing fructose 1, 6-biphosphatase 1 (FBP1), and to investigate whether FBP1 has effect on autophagy and proliferation in liver cancer cells (HepG2).@*Methods@#FBP1 cDNA sequence was amplified by PCR and cloned in adenovirus vector pAdTrack-TO4, and then recombinant adenovirus plasmid pAdTrack-FBP1 was constructed. The recombinant adenovirus plasmid was transfected into HEK293 cells by Lipofectamine 3000. High-titer of recombinant adenovirus AdFBP1 was obtained by packaging and amplification. HepG2 cells were infected with recombinant adenovirus AdFBP1, and the Mock and AdGFP group were set at the same time. Western blot and confocal laser scanning microscopy were used to observe the effect of FBP1 on the level of autophagy in hepatocellular carcinoma cells, and the effect of FBP1on the proliferation was observed by MTS and colony formation assay. A t-test and one-way ANOVA were used to compare the mean between group.@*Results@#A high-titer recombinant adenovirus FBP1 was successfully constructed. Western blot and confocal laser scanning microscopy showed that the level of autophagy in AdFBP1 group was significantly lower than that in AdGFP group. Western blot results showed that LC3-II protein expression level in AdGFP was 1.10 ± 0.10 and 0.30 ± 0.01 in AdFBP1 group, F = 90.36, P < 0.01. Confocal laser scanning microscopy analysis showed that the average number of autophages in AdGFP was 28.33 ± 1.53 and 12.33 ± 1.53 in AdFBP1group, F = 97.40, P < 0.01. In addition, the results of colony formation assay and MTS assay showed that the proliferation of liver cancer cells in the AdFBP1 group was significantly inhibited compared with the AdGFP group. The results of colony formation showed that the cell clones in the AdGFP group was 65.66 ± 2.57 and 34.00 ± 2.00 in AdFBP1 group, F = 141.50, P < 0.01. MTS results showed that the absorbance of AdGFP group at 96h was 39.13 ± 2.21 and 30.61 ± 3.33 in AdFBP1 group, F = 7.80, P < 0.05.@*Conclusion@#FBP1 inhibited the autophagy and proliferation in liver cancer cells (HepG2).
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OBJECTIVE: To analyze the clinical and molecular genetic characteristics of 2 cases of fructose-1,6-bisphosphatase deficiency in the same family to provide evidence for the precise treatment,genetic counseling and prenatal diagnosis.METHODS: Clinical data were collected from 2 patients with hypoglycemia encephalopathy,and molecular genetic analysis was performed using targeted capture next-generation sequencing. RESULTS: The 2 patients were siblings,the male proband was 7 years old,mainly manifested with convulsions after hunger or ingestion of a large amount of fructose,accompanied by ketoacidosis;clinical diagnosis was hypoglycemia encephalopathy,and fructose metabolism abnormalities was suspected. The younger brother was 4 years old,mainly showing hunger and sweating in the morning,stomach ache after eating fruit,and convulsion episode once after hunger. Next-generation sequencing results showed that the siblings had c.333+1＿2 delinsTC and c.490 G>A compound heterozygous mutations in the FBP1 gene,and their parents were carriers with normal phenotype.The c.333+1＿2 delins TCis a novel mutation,c.490 G>A is a reported pathogenic mutation,and the two patients were diagnosed with fructose-1,6-bisphosphatase deficiency genetically. CONCLUSION: For children with unexplained hypoglycemia,convulsions and metabolic acidosis,the fructose-1,6-bisphosphatase deficiency should be considered. Early genetic analysis is helpful to clarify the cause,make precise treatment and improve prognosis.
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Objective: To study the inhibitory effect of the medicine group of promoting blood circulation and removing stasis (PBCRS) on breast cancer induced by 7, 12-dimethylbenz(a)anthracene (DMBA) in rats, and screen out and verify key genes based on RNA Sequencing (RNA-seq) technology and Ingenuity Pathway Analysis (IPA). Method: Totally 96 Sprague-Dawley (SD) rats were randomly divided into blank group, DMBA model control group, tamoxifen (TAM) group (1.9 mg·kg-1·d-1), high-dose, middle-dose and low-dose PBCRS groups (12.96, 6.48, 3.24 g·kg-1·d-1). One week after drug intervention, except for the blank group, the DMBA was used to induce the rat model of breast cancer (with an interval of a week, irrigation for two times at the dose of 100 mg·kg-1). After 10 weeks, the changes in tumor weight and tumor volume were observed. The total RNA was extracted by total RNA extraction kit, and three RNA samples were collected from the DMBA model control group and the middle-dose PBCRS group for genetic testing. Based on RNA-seq, key differential genes were screened out and verified by Real-time PCR. Result: Comparing with the DMBA model control group, the tumor volume and tumor weight in middle-dose PBCRS group were decreased significantly (PPConclusion: PBCRS may inhibit the occurrence of breast cancer by interfering with the expression of FBP1 in breast cancer tissue.
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@#[Abstract] Objective: Toevaluatetheexpressionof6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3(PFKFB3) in malignant glioma tissues and the effects of inhibitor of PFKFB3(PFK15) on the proliferation, migration, invasion, clone formation and tumorigenesis of H4 cells. Methods: Malignant brain glioma tissues and corresponding paratumor tissues from 31 patients, who were hospitalized in Department of Neurosurgery,Ankang Hospital of Traditional Chinese Medicine during February 1, 2015 to January 31, 2016 for operative treatment, were collected for this study. Immunohistochemistry and western blotting assays were applied to detect the expression of PFKFB3 in collected tissues. PFKFB3 in H4 cells were blocked by PFK15 (1.25, 2.5, 5.0 μmol/L). The effect of PFK15 on proliferation, migration, clone formation and tumorigenesis of H4 cells were determined by MTT assay, EdU incorporation assay, wound healing assay, Transwell assay, colone formation assay and in vivo xenograft bearing nude mice model respectively. Results: Positive expression rate of PFKFB3 was significantly higher in malignant glioma tissues compared with normal adjacent tissues[(80.60±8.98)% vs (41.57±10.16)%, P<0.05]. The results of MTT assay and EdU incorporation assay indicated that PEK15 significantly inhibited the proliferation of H4 cells in a concentration dependent manner. The migration, invasion and clone formation activity of H4 cells were significantly reduced by treatment with PFK15 (all P<0.05). In tumor bearing nude mice, the tumor volume of mice treated with PFK15 was significantly smaller than that of mice from control group ([254.15±154.25] vs [801.52±224.25] mm3, P<0.05). Conclusion: PFKFB3 was highly expressed in malignant glioma tissues. Blocking of PFKFB3 by PFK15 significantly reduced the malignant biological behaviors and tumorigenesis of H4 cells in vitro and in vivo, which may serve as a promising target for the treatment of malignant gliomas.
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During 2 years,a 6-year-old girl was hospitalized for 2 times with recurrent onset of episodes of vomiting,weakness and fever after eating dessert at the Department of Neurology & Endocrine Pediatrics,the Affiliated Hospital of Qingdao University.The arterial blood gas analysis revealed severe hypoglycemia,lacticacidemia and metabolic acidosis,the urine ketone body was positive.After intravenous infusion of glucose,bicarbonate and antibiotics,there was a dramatic clinical improvement in a short time.Physical examination showed tachypnea and mild hepatomegaly,and she had normal physical and mental development.The laboratory findings revealed transient hyperuricacidemia.Urine organic acids analysis repeatedly showed an elevation of lactic acid,ketone and glycerol.Glyceroluria was a very distinctive trait.The literatures in PubMed was searched with glyceroluria as keyword.Three related diseases were identified:FBPase deficiency,glycerol kinase (GK) deficiency and complex GK deficiency.Further reading of related literatures to understand the characteristics of diseases and laboratory tests,the clinical diagnosis of GK deficiency and complex GK deficiency was excluded.The mutation analysis of FBPase gene (FBP1) was performed by Sanger sequencing and a novel compound heterozygous mutations of c.355G ＞A and c.960delG was discovered.Full analysis of disease-related traits and targeted gene testing is one of the effective methods for accurate diagnosis and treatment of inherited metabolic disorders.
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Objective To investigate the expression of FBP1 in human renal clear cell carcinoma and paracancerous tissue and its effect and clinical significance in the carcinogenesis,progression and prognosis of renal cancer.Methods The paraffin sections from 118 patients with renal clear cell carcinoma treated by surgical resection and fresh specimens from 40 patients with renal clear cell carcinoma were selected.The expressions of FBP1 protein and mRNA in renal cancer and paracancerous tissues(negative incisal edge) were detected by adopting the immunohistochemistry(IHC),Western blot and RT-PCR.Its correlation with clinicopathologic characteristics and prognosis of the patients was analyzed.Results The IHC result,found that strong positive expression of FBP1 protein could be seen in 78.81％ (93/118) of cancer-adjacent tissues,while only 39.83％ (47/118) of renal cancer tissues had positive expression.Western blot found that the expression positive rate of FBP1 in renal cancer tissue was significantly decreased compared with corresponding cancer-adjacent tissues (P＜0.01).RT-PCR found that the FBP1mRAN expression level in cancer-adjacent tissues was also significantly higher than that in renal cancer tissue(P＜0.05).The FBP1 low expression was significantly correlated with the clinical stage,pathologic grade,UISS risk coefficient and recurrence(P＜0.05),and had no relation with the age,gender,symptoms,tumor size,location,tumor necrosis,vascular invasion and adrenal involvement(P＞0.05).The 5-year survival rate in renal cancer patients with FBP1 positive expression was higher than that in the patients with FBP1 negative expression(P＜0.05).Conclusion FBP1 and protein are lowly expressed in renal cancer tissue,are correlated with occurrence and development of renal cancer,and may become one of candidate markers of renal cancer prognosis.
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Background and purpose: Epithelial ovarian carcinoma is the most malignant tumor in female reproductive system because of its resistance to chemotherapy. Fructose-1, 6-bisphosphatase (FBP1) is a rate-limiting enzyme in gluconeogenesis used to catalyze the hydrolysis of fructose-1, 6-bisphosphate to fructose-6-phosphate and inorganic phosphate, thereby inhibiting the effect of glycolysis in tumor cells. This study aimed to investigate the association between the expression of FBP1 and chemosensitivity. Methods: The expression level of FBP1 in ovarian cancer patients was measured by immunohistochemistry. Results: According to the results of immunohistochemistry in 209 ovarian carcinoma specimens, the percentage of positive FBP1 expression was about 49.3% (103/209). Loss of FBP1 was a negative factor of survival (42.6 months vs 62.1 months, P=0.003). Besides, patients who were sensitive to chemotherapy displayed significantly higher scores of FBP1 expression than patients who were resistant to therapy (P=0.007). Conclusion: The rate-limiting enzyme FBP1 in gluconeogenesis can be used as a biomarker for predicting the chemoresistance and prognosis of ovarian cancer patients.
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Metabolic rearrangement is a typical hallmark of cancer cells ,especially the disorders in glucose metabolism. Normal cells rely on the steps of oxidative phosphorylation within the mitochondria to metabolize glucose and yield energy upon enough oxygen ,yet turning to glycolysis in the absence of oxygen . Nevertheless ,tumor cells exhibit high levels of glycolytic flux in different environments ,due to the requirement of malignant proliferation. This phenomenon was named as the Warburg effect ,the initiation of which is driven by multiple mechanisms ,including the influence of tumor microenvironment ,the activa?tion of tumor-promoting signaling pathways ,and the aberrant levels of metabolic enzymes. Moreover ,our recent findings sug?gest that inhibition of gluconeogenesis ,the general reversal of glycolysis ,would further promote the Warburg effect and tumor progression.
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Fructose-1,6-bisphosphatase(FBPase) deficiency is a rare inherited metabolic disease,which is an autosomal recessive metabolic disorder.Affected patients usually present with metabolic crisis including severe hypoglycemia and metabolic acidosis.Each attack occurred with a similar sequence.The triggering factors are removed and then clinical status is improved dramatically.As patients are usually symptomless in the plateau stage,it is often misdiagnosed.Metabolite assay in blood and urine is very useful for the diagnosis of FBPase deficiency.FBPase is a key enzyme in gluconeogenesis.Deficiency of FBPase impairs the formation of glucose from all precursors.FBP1 gene mutation contributes to the disease.More than 30 mutation types have been reported.There is no specific treatment.Early diagnosis and appropriate life-style can prevent repetitive metabolic derangements,improving life quality of these children and ensuring successful pregnancy.
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The gene mutations of a patient with fructose-1,6-bisphosphatase (FBPase) deficiency and her parents were analyzed and her clinical manifestations, laboratory tests, and genetic characteristics were reviewed. The molecular analysis of FBP1 gene showed a G residue duplication at base 960 in exon 7(c. 960dupG) in this patient while her parents carried the heterozygous c. 960dupG mutation. The prominent clinical feature of this patient was the benign course of the disease with age. However, acute attack could be triggered by stress, long-time fasting, a large amounts of fructose intake, etc. The typical clinical manifestations were severe lactic acidosis, hypoglycemia, and elevated liver enzymes.
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The gene mutations of a patient with fructose-1,6-bisphosphatase (FBPase) deficiency and her parents were analyzed and her clinical manifestations, laboratory tests, and genetic characteristics were reviewed. The molecular analysis of FBP1 gene showed a G residue duplication at base 960 in exon 7(c. 960dupG) in this patient while her parents carried the heterozygous c. 960dupG mutation. The prominent clinical feature of this patient was the benign course of the disease with age. However, acute attack could be triggered by stress, long-time fasting, a large amounts of fructose intake, etc. The typical clinical manifestations were severe lactic acidosis, hypoglycemia, and elevated liver enzymes.