Effect of amitriptyline on lipid deposition and biochemical metabolism in a cell model of nonalcoholic fatty liver disease / 临床肝胆病杂志

ObjectiveTo investigate the effect of amitriptyline on lipid deposition and biochemical metabolism in a cell model of nonalcoholic fatty liver disease (NAFLD) by regulating the acid sphingomyelinase (ASM)/ceramide (CE) pathway. MethodsHepG2 and L02 cells were cultured in vitro to establish a cell model of NAFLD. MTT colorimetry was used to measure cell proliferation rate, and oil red O staining was used to observe the change of lipid droplets in cells. In the experiment, the cells were divided into normal control group, model group, Ami group, TNFα group, and Ami+TNFα group. An automatic biochemical analyzer was used to measure the levels of triglyceride (TG) and total cholesterol (TC) in cells and the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in supernatant; ELISA was used to measure the levels of CE and ASM in cells; Western blot was used to measure the protein expression ASM in cells, and RT-PCR was used to measure the mRNA expression of ASM in cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the Turkey test was used for further comparison between two groups. ResultsCompared with the normal control group, the NAFLD model group had significant increases in the protein and mRNA expression of ASM and the levels of CE, TG, TC, ALT, and AST (all P＜0.05). Compared with the model group, the Ami group had significant reductions in the protein and mRNA expression of ASM and the levels of CE, TG, TC, ALT, and AST (all P＜0.05), and the TNFα group had significant increases in the protein and mRNA expression of ASM and the levels of CE, TG, ALT, and AST (all P＜0.05). Compared with the TNFα group, the Ami+TNFα group had significant reductions in the protein and mRNA expression of ASM and the levels of CE, TG, TC, ALT, and AST (all P＜0.05). ConclusionThe ASM/CE pathway promotes lipid accumulation and may lead to hepatocyte steatosis, and amitriptyline can alleviate lipid deposition in NAFLD hepatocytes by inhibiting the ASM/CE pathway.

Imaging diagnosis of hepatic lipidosis in a cat / 대한수의학회지

A 5-year-old intact female Persian cat was referred with anorexia and vomiting. Physical examination revealed icterus in the pinna, oral mucosa, and sclera. Ultrasonography revealed several findings including uniformly hyperechoic hepatic parenchyma compared to adjacent falciform fat, increased attenuation of the ultrasound beam, and poor visualization of intrahepatic vessel borders. Computed tomography revealed hypoattenuation of the hepatic parenchyma with a radiodensity value of

Influences of high-fat diet during pregnancy on sterol regulatory pathway and hepatic lipid deposition in male rat offspring / 中华围产医学杂志

Objective To analyze the influences of maternal high-fat diet on male rat offspring's blood lipid level and hepatic lipid deposition as well as on the expression of two key factors,sterol regulatory element binding protein-1 (SREBP-1) and SREBP cleavage acting protein (SCAP),involved in hepatic cholesterol regulatory cascade during childhood and adulthood.Methods Pregnant Wistar rats were randomly divided into two groups:high fat diet (HF) group and normal intake control (NC) group (both n=20).Rats in the HF group were fed with high-fat diet till delivery before changing to a normal-fat diet,while the NC group was given the normal-fat diet all through the study.Male offspring born appropriate for gestational age (AGA) in different feeding groups were randomly selected as experimental subjects.Physical development,serum lipid levels and hepatic lipid deposition at the age of 7 and 24 weeks were compared between offspring of the two groups.Expression of SCAP and SREBP-1 at protein level was detected with immunohistochemistry.T test or Chi-square test was used for statistical analysis.Results The average birth weight of HF group was higher than that of NC group [(6.95± 0.25) vs (6.79 ±0.78) g,t=2.088,P=0.038].Large for gestational age (LGA),AGA and small for gestational age (SGA) offspring accounted for 18.6％ (21/113),77.9％ (88/113)and 3.5％ (4/113) in HF group and 7.6％ (10/132),87.9％ (116/132) and 4.5％ (6/132) in NC group (x2=13.500,P=0.001).At the age of 7 weeks,the offspring's body weight in HF group was higher than that in NC group [(68.78±7.55) vs (66.61 ±3.92) g,t=2.303,P=0.023].At the age of 24 weeks,the body weight,body length and abdominal circumference of adult offspring in HF group were significantly higher than those in NC group [(251.74±24.04) vs (216.24±33.42) g,(22.60±0.79) vs (21.59± 1.34) cm,(17.93±0.59) vs (16.83±0.88) cm;t=2.220,2.379,2.927,all P＜0.05].At the age of 7 weeks,offspring's serum total cholesterol (TC) level in HF group was higher than that in NC group [(1.94±0.62) vs (1.23 +0.77) mmol/L,t=2.379,P＜0.05].At the age of 24 weeks,adult offspring's liver index,serum TC,triglyceride (TG),low density lipoprotein-cholesterol (LDL-C) and very low density lipoprotein-cholesterol (VLDL-C) levels were all higher in HF group than in NC group,while the high density lipoprotein-cholesterol (HDL-C) level was lower in HF group [(2.75±0.31)％ vs (2.46±0.39)％,(1.98±0.14) vs (1.45±0.27) mmol/L,(2.96±0.21) vs (2.71 ±0.29) mmol/L,(0.97±0.32) vs (0.71 ±0.21) mmol/L,(0.81 ±0.13) vs (0.52±0.17) mmol/L,(1.74±0.20) vs (1.59±0.29) mmol/L;t=2.264,2.333,2.359,2.088,5.336,1.994,all P＜0.05].Comparative analysis of nonalcoholic fatty liver disease activity scores (NAS) between the two groups showed that the liver lobular inflammation in HF group's offspring was more severe at both 7 and 24 weeks of age (2.29±0.49 vs 1.92±0.45,2.83±0.47 vs 2.22±0.55;t=2.157,3.251,all P＜0.05).At the age of 24 weeks,adult offspring in HF group also showed higher liver steatosis scores (2.81 ± 0.35 vs 2.25 ± 0.30,t=4.609,P＜0.05).Immunohistochemical detection showed that there was no significant difference in SCAP protein expressions in liver tissues of 7-week-old offspring between the two groups (P＞0.05),but higher expression of SREBP-1 protein was observed in HF group than in NC group [(34.16±5.08)％ vs (18.09±3.99)％,t=9.697,P＜0.05].At the age of 24 weeks,the expression levels of both SCAP and SREBP-1 proteins in liver tissues were higher in HF group than in NC group [(31.22±6.01)％ vs (17.98+7.89)％,(61.33± 16.25)％ vs (29.76± 11.21)％;t=2.303,2.274,both P＜0.05].Conclusions High-fat intake during pregnancy can increase offspring's birth weight and the risk for LGA offspring.Maternal high-fat diet even up-regulates the expression of SCAP and SREBP-1 in male offspring born AGA in childhood and this impact become more obvious when reaching adulthood,resulting in excessive increase of body weight,body length and abdominal circumference and elevated serum lipid level and liver lipid deposition.

Neutral lipid storage disease with distal weakness: report of one family / 中华神经科杂志

Objective To report the clinical and pathological features of one family with neutral lipid storage disease with distal weakness and review the literature , explore the diagnosis and differential diagnosis of the disease , and improve the understanding of the disease .Methods Clinical data of two patients of a family were collected , and next generation sequencing was performed after electrophysiological examination and pathological analysis of skeletal muscle biopsy .And related literature was reviewed .Results Two patients had similar presentations , both having symptoms after 30 years old.The main manifestations were weakness and atrophy of the distal limbs .The creatine kinase levels were increased ( 1067, 740 U/L) .Electromyography revealed myogenic lesions .Pathological analysis of skeletal muscle biopsy showed a large amount of lipid droplets deposition in most muscle fibers .Next generation sequencing revealed a homozygous splice mutation in PNPLA2 gene ( C.187 +1G >A) .Conclusions The two patients with neutral lipid storage disease with myopathy involve distal limbs , which should be distinguished with distal myopathy.The presence of large amounts of lipid droplets in the cytoplasm of muscle fibers can be used for the diagnosis of lipid storage myopathy .The next generation sequencing is helpful for the typing diagnosis of lipid deposition disease .

Quantification of Organ Fat Deposits in Patients with Type 2 Diabetes Mellitus by Magnetic Resonance Imaging / 中国医学影像学杂志

Purpose Lipotoxicity plays an important role in the progression of diabetes and its complications,and the gold standards for organs' lipid quantification are biopsy or histochemical staining,which are invasive and have their own limitations.Thus,our research was to identify the difference in organ lipid deposition between type 2 diabetic patients and healthy volunteers by using noninvasive three-point T2* corrected Dixon imaging.Materials and Methods Sixty-five type 2 diabetic patients and 34 healthy volunteers were included in this study.All participants underwent three-echo Dixon with 3.0T MR imager.Dixon imaging parameters [hepatic fat fraction (HFF),splenic fat fraction (SFF),pancreatic fat fraction (PFF)] were collected.All of the MRI parameters were compared.The relationship between HFF,SFF,PFF and BMI,age were analyzed.Results The hepatic and splenic lipid percentage in diabetic group [HFF=(5.4±4.3)％;SFF=(3.7± 1.4)％] was significantly higher than healthy volunteers group [HFF=(2.9± 1.3)％;SFF=(3.0± 0.9)％;P＜0.05,respectively].However,there was no significant difference in pancreatic fat fraction between diabetic and healthy groups (P＞0.05).There were positive correlations between HFF,SFF,PFF and BMI (r=0.379,0.305 and 0.306,P＜0.05).Moreover,only the positive correlation between pancreatic fat fraction and age were observed (r=0.261,P＜0.05).Conclusion The three-point T2* corrected Dixon revealed the abnormalities of hepar and spleen lipid accumulation in diabetic patients.Thus,the three-point Dixon imaging may potentially aid in evaluating the lipid deposition of abdominal organs.

Influence of rotation manipulation on Raman spectral characteristics of stable plaques in a rabbit model of early carotid atherosclerosis / 中国组织工程研究

BACKGROUND:It is generaly recognized that cervical rotation manipulation can increase the risk of detachment of unstable plaques in carotid atherosclerosis, but few studies are reported on the influence of cervical rotation manipulation on the stable plaque in early carotid atherosclerosis. OBJECTIVE:To explore the influence of the cervical rotation manipulation on the lipid contents in carotid atherosclerotic plaque in a rabbit model of early carotid atherosclerosis. METHODS:After being fed for 15 days with normal diet, 30 male New Zealand White rabbits were further fed for 18 weeks with normal diet (n = 10; control group) or a high-fat diet containing 2% cholesterol, 10% lard and 88% normal granules to build rabbit models of early carotid atherosclerosis with stable carotid plaque (n = 20). At 14 weeks of feeding with high-fat diet, the experimental rabbits fed with high-fat diet were randomly divided into a cervical rotation manipulation group (n = 10) and a model group (n = 10). The rabbits in the cervical rotation manipulation group underwent cervical rotation manipulation to the left and right sides, once each side, with the maximal range of rotation. Total five cervical rotation manipulations, once every 3 days, were performed. RESULETS AND CONCLUSION:The 1 450 cm-1 and 1 660 cm-1 peaks of the Raman spectrum of lipid in the carotid atherosclerotic plaque of rabbit models were not distinctly present in the control group, however, they were obviously observed in the cervical rotation manipulation group and model group. Nevertheless, the relative intensity differences at spectrum characteristic peaks were not significant between cervical rotation manipulation group and model group (P > 0.05). The environmental findings indicate that the lipid content in carotid atherosclerotic plaque of rabbit models of early carotid atherosclerosis cannot be increased after short-term administration of cervical rotation manipulation.

Hot spot mutations in electron transfer flavoprotein dehydrogenase gene of riboflavin responsive lipid storage myopathy in 20 Chinese families / 中华神经科杂志

Objective To report the spectrum of electron transfer flavoprotein dehydrogenase (ETFDH)gene mutations in 20 Chinese RR-LsM families.Methods Twenty-four RR-LSM patients in the First Hospital of Peking University from January 2003 to May 2010 were collected and the clinical characteristics were analyzed.These patients came from 20 families in North Mainland China.Sixteen families had 1 patient each.and the other 4 families had 2 patients.ETFDH gene analysis was performed in all patients,11 family members and 100 healthy controls.Results The mean onset age was(27.9±9.9)years.The main symptoms were limb weakness(21,87.5%),dysmasesia(15,62.5%),neck weakness (14,58.3%)and myalgia(14,58.3%).Eighteen patients had high level of acyleamitine.Fifteen of 17patients had glutaric aciduria.Seventeen ETFDH mutations,including 13 missense mutations,2 splice mutations,and 2 nonsense mutations,were identified in 19 families:c.998A>G,c.1450T>C,c.1703T>C,c.1717C>T,c.821G>A,c.643G>A,c.251C>T,c.1763A>T,c.IVS7+2T>C and c.IVS6+1G>A were Hovel mutations which were not found in 100 healthy controls.Nine families had the mutation of c.770A>G(P.Y257C)and 5 families had the mutation of c.1227A>C(P.L409F).Conclusions The numerous novel mutations in ETFDH gene indicate that Chinese RR-LSM might have special mutation pattern.c.770A>G(P.Y257C)and c.1227A>C(p.L409F)may be hot spot mutations in North Mainland China.

Clinical features and electron transfer flavoprotein dehydrogenase gene mutation analysis in 35 Chinese patients with lipid storage myopathy / 中华神经科杂志

Objective To investigate the clinical features and electron transfer flavoprotein dehydrogenase (ETFDH) gene mutations in 35 Chinese patients with lipid storage myopathy. Methods The clinical data of 35 cases with lipid storage myopathy confirmed by muscle biopsy were collected. The sequences of all 13 exons of ETFDH were analyzed. Results All 35 patients showed proximal weakness. Ten of them demonstrated masseter weakness and 28 of them showed weakness in neck flexion. Twenty-nine of 32 patients who were followed up showed improvement after treatment with VitB2 and CoQ10. Mutations of ETFDH were found in 30 of 35 patients,which included 8 homozygosises,20 compound heterozygosises and 2 single heterozygosises. Fourteen novel mutations were found, including 9 missense mutations ( c. 3G > C, c. 152G>A, c. 191G > A, c.349G>C, c.433G>C, c. 949C > A, c. 1454C > G, c. 1744A >T and c. 1763A>G), 1 nonsense mutation(c. 172G>T), 2 deletions(c. 1282_1283del and 1773_1774del) and 2 splice mutations (c. 405 + 1G > T and c. 1691 -3C > G). Nine of them showed c. 250G > A mutation and 6 of them showed c. 770A > G mutation. Conclusions Lipid storage myopathy is presented as proximal weakness. Multiple acyl-CoA dehydrogenase deficiency caused by mutations of ETFDH is the major cause of lipid storage disease in this group. ETFDH c. 250G > A and c. 770A > G mutations show a high frequency.

Estudo da doença de Gaucher em Santa Catarina / Study of Gaucher disease in Santa Catarina

A doença de Gaucher (DG) foi a primeira doença de armazenamento lisossomal descrita e a mais encontrada. Caracteriza-se pela deficiência hereditária da atividade da enzima lisossomal glucocerebrosidase, que bloqueia o metabolismo do glicocerebrosídeo. A proposta deste trabalho foi estudar as características clínicas, laboratoriais e radiológicas, as principais mutações encontradas, relacionando-as com as formas clínicas e avaliar a resposta à terapia de reposição enzimática (TRE) nos pacientes com DG em Santa Catarina. Foram estudados dez pacientes com DG no Hospital Universitário, no período entre 1998 e 2003, após confirmação diagnóstica da doença pela dosagem da enzima beta-glicosidase em leucócitos. Pesquisa das mutações foi realizada em amostras de sangue e de mucosa oral. A média de idade ao diagnóstico foi de 19,6 anos. A DG tipo 1 foi diagnosticada em 80 por cento dos casos, e a tipo 2 em 20 por cento. Quatro pacientes tiveram história familiar de DG. Hepatoesplenomegalia foi a manifestação clínica mais comum. Anemia e trombocitopenia ocorreram em 100 por cento dos casos. Dores ósseas foram relatadas por 75 por cento dos pacientes. Os alelos mutantes encontrados foram N370S e L444P. Houve elevação dos níveis de hemoglobina em todos os pacientes com DG tipo 1. Concluímos que a DG tipo 1 é a forma clínica mais comum. Anemia, trombocitopenia, hepatoesplenomegalia e osteopenia são as características mais freqüentes dos pacientes com DG. O alelo N370S é o mais freqüente, estando relacionado com o tipo 1. O alelo L444P em homozigose sugere letalidade precoce. A TRE é segura e efetiva para a DG tipo 1.

Gaucher Disease (GD) was the first described and is the most common lysosomal deposit disease. It is characterized byahereditary deficiency of glucocerebrosidase lysosomal enzyme activity which blocks the metabolism of glucocerebrosideo. The aim of this work was to study the clinical, laboratorial and radiological characteristics, the main mutations correlating them with the clinical form of the disease and evaluating the response to enzymatic replacement therapy in patients with GD in Santa Catarina. Ten GD patients were studied at a University Hospital between 1998 and 2003. The disease was diagnosed by measurement of the beta-glucosidase enzyme in leukocytes. Investigation of mutations used samples of blood and oral mucus. The average age at diagnosis was 19.6 years. Type 1 GD was diagnosis in 80 percent of the cases and type 2 in 20 percent. Four patients had a family history of GD. Hepatosplenomegaly was the most common clinical manifestation. Anemia and thrombocitopenia occurred in all cases. Bone pain was reported by 75 percent of the patients. The mutanted alleles identified were N370S and L444P. The hemoglobina levels were elevated in all patients with type 1 GD. In conclusion, type 1 GD is the most common clinical form and anemia, thrombocitopenia, hepatosplenomegaly and osteopenia were the most common characteristics of GD patients. The N370S allele is the most common mutation and is related with type 1 GD 1. Homozygosity of the L444P allele suggests early death. Enzyme replacement therapy is safe and efficacious in type 1 GD.

Síndrome de Urbach-Wiethe: relato de caso / Urbach-Wiethe syndrome: a case report

Apresentação de um caso de síndrome de Urbach-Wiethe com manifestações típicas, inclusive oculares. Paciente do sexo feminino, 15 anos, com quadro de prurido ocular relacionado à presença de lesões papuliformes em margens palpebrais (blefarose moniliforme), em associação com outras alterações sistêmicas. O diagnóstico foi confirmado por meio de biópsia cutânea e foi instituído uso de lágrimas artificiais, com alívio parcial do sintoma. O objetivo do trabalho é relatar um caso com manifestações características da doença atendido no Setor de Oftalmologia da Faculdade de Medicina de Marília.

We present a case of Urbach-Wiethe syndrome with typical findings, including ocular lesions. A 15-year-old girl was referred to our department complaining of itchy eyelid lesions (moniliform blepharosis) associated with other systemic manifestations. Diagnosis was confirmed by performing skin biopsy. Artificial tears were prescribed, with partial relief of the symptom. The objective of the present study is to describe a typical case of Urbach-Wiethe syndrome attended at the Ophthalmologic Sector of the Medical School of Marília.