Objective To investigate the effect and mechanism of S100A16 in the course of 3T3-L1 preadipocytes differentiation.MethodsA recombinant virus vector overexpressing S100A16 ( PLJM1-S100A16-GFP) was constructed and transfected into 3T3-L1 preadipocytes.The expression of S100A16 in the course of 3T3-L1 preadipocytes differentiated into adipocytes was detected by Western blot.The lipid droplets were observed by oil-red O staining and triglycercide (TG) content was measured by the TG measure kit.Levels of adipogenesis-associated genes such as PPARγ,CCAAT/enhancer binding protein ( C/EBP-α),lipoprotein lipase ( LPL),fatty acid synthase ( FAS),adipocyte fatty acid binding protein( aP2 ) were measured by realtime PCR and Western blot.The interaction between S100A16 and p53 was detected by immunoprecipitation.Results3T3-L1 cell line overexpressing S100A16was successfully contructed.It was found that the expression of S100A16 was increased during 3T3-L1 adipocytes differentiation.Overexpression of S100A16 stimulated 3T3-L1 preadipocytes differentiation and increased the accumulation of triglycerides in adipocytes (P＜ 0.01 ),along with the up-regulation of adipocyte differentiationassociated gene expressions including PPARγ,C/EBP-α,LPL,aP2,and FAS ( P ＜ 0.05 or P ＜ 0.01 ).Immunoprecipitation analysis revealed that S100A16 interacted with tumor suppressor protein p53,also a known inhibitor of adipogenesis.ConclusionS100A16 stimulates 3T3-LI preadipocytes differentiation via inhibiting p53activity.

Objective:To describe a method based on analysis of the histogram of intensity values pro-duced from the magnetic resonance imaging (MRI)for quantifying the degree of fatty infiltration. Methods:The study included 25 patients with dystrophinopathy.All the subjects underwent muscle MRI test at thigh level.The histogram Mvalues of 250 muscles adjusted for subcutaneous fat,representing the degree of fatty infiltration,were compared with the expert visual reading using the modified Mercuri scale.Results:There was a significant positive correlation between the histogram Mvalues and the scores of visual reading (r =0.854,P <0.001).The distinct pattern of muscle involvement detected in the pa-tients with dystrophinopathy in our study of histogram M values was similar to that of visual reading and results in literature.The histogram M values had stronger correlations with the clinical data than the scores of visual reading as follows:the correlations with age (r =0.730,P <0.001 )and (r =0.753, P <0.001);with strength of knee extensor (r =-0.468,P =0.024)and (r =-0.460,P =0.027) respectively.Meanwhile,the histogram Mvalues analysis had better repeatability than visual reading with the interclass correlation coefficient was 0.998 (95% CI:0.997 -0.998,P <0.001)and 0.958 (95%CI:0.946 -0.967,P <0.001)respectively.Conclusion:Histogram Mvalues analysis of MRI with the advantages of repeatability and objectivity can be used to evaluate the degree of muscle fatty infiltration.

Objective To investigate the synergistic effect of 11 β-hydroxysteriod dehydrogenase (11 β-HSD1) and S100A16 on the differentiation of3T3-L1 preadipocytes and its mechanism.Methods Lentiviral vectors PLJM1-11β-HSD1 and PLJM1-S100A16-GFP were respectively constructed and co-transfected into 3T3-L1 preadipocytes.The cell strains expressing 11 β-HSD1/S100A16 were screened with 2.5 μg/ml puromycin for two weeks.Western blot was employed to verify the lentiviral carrier transfection effects.The expressions of marker genes related to the adipocyte differentiation were detected by mean of realtime PCR.Oil red O staining was used to observe the lipid droplet accumulation and the content of triglyceride was measured after differentiation of preadipocytes.The effect of 11β-HSD1 and S100A16 on PPARγ promoter activity was detected by luciferase reporter gene.Results Compared with the empty vector group,the expressions of 11β-HSD1 and S100A16 protein in the lentivirus cotransfected 3T3-L1 cell strain were significantly higher.After 3T3-L1 cell strain co-expressing 1 1β-HSD1 and S100A16 was induced to differentiate for 8 days,the lipid droplets accumulation and triglyceride content were siginificantly increased,along with increased expressions of adipocyte differentiation marker genes such as PPARγ,CCAAT/enhancer binding protein α,lipoprotein lipase,fatty acid synthase,and adipocyte fatty acid-binding protein,in comparison with 11 β-HSD1 or S100A16 overexpression.The result of reporter gene indicated that 11 β-HSD1/ S100A16 enhanced PPARγ promoter activity.Conclusions 11β-HSD1 and S100A16 may jointly promote the differentiation of 3T3-L1 preadipocytes through a synergistic effect on PPARγexpression and play a critical role in the development of obesity.

Objective To investigate the characteristics of muscle edema and fatty infiltration in thighs and relationship with clinical symptoms in Chinese patients with different phenotypes of dysferlinopathy.Methods A total of 32 patients were enrolled , including 13 limb-girdle muscular dystrophy 2B (LGMD2B), 13 Miyoshi myopathy (MM), 4 proximodistal myopathy and 2 hyper-creatine-kinase-emia.Clinical symptoms were evaluated using modified Gardner-Medwin and Walton ( GM-W) score.Muscle MRI was performed in thighs to observe fatty infiltration and edema.We then compared the age of onset , disease duration, GM-W score, muscle edema and muscle fatty infiltration between LGMD 2B and MM groups,and the relationship of muscle edema score and fatty infiltration score with disease duration and GM-W score in all patients.Results The median GM-W score was 4.00 (2.00,5.00) in all patients, 4.00 (3.00,4.50)in LGMD2B and 4.00(2.00,5.00)in MM, respectively.Muscle fatty infiltration appeared in 30 cases (93.75%), with the same pattern in LGMD2B and MM.The mean fatty infiltration score was 28.20 ±12.86 in all patients, 28.50 ±13.03 in LGMD2B and 29.00 ±12.63 in MM, respectively.Muscle edema appeared in 26 cases (81.25%) with same pattern in LGMB2B and MM.The mean edema score was 18.36 ±13.60 in all patients, 22.88 ±11.59 in LGMD2B and 16.77 ±13.80 in MM.The age of onset , disease duration, GM-W score, muscle fatty infiltration and edema score were not significantly different between LGMD2B and MM patients.Muscle fatty infiltration score significantly correlated with GM-W score (rs=0.737,P=0.000) and disease duration (rs=0.637,P=0.000).Conclusions Fatty infiltration and edema in thigh muscles are very common in patients with dysferlinopathy , with similar radiological changes in main subtypes.The muscle fatty infiltration can be used as a predictor of disease progression.

Objective To analyze the preventive and therapeutic effects of a traditional Chinese medicine Taohe Chengqi decoction on a rat models of hepatic encephalopathy ( HE) induced by thioacetamide ( TAA) .Methods The rat model of HE was established by intraperitoneal injection of TAA,and the effects of Taohe Chengqi decoction were observed by neurobehavioral changes,neurological test,blood ammonia,serum biochemical indexes,and liver and brain pathological examination.Results The rat model of hepatic encephalopathy was successfully established.Low, medium and high dose Taohe Chengqi decoction markedly improved neuro-reflexes and liver and brain pathological damages, and significantly decreased the HE staging and serum biochemical indexes, and the results of high dose group was the best, similar to that of positive drug-treatment group.Conclusions Taohe Chengqi decoction exhibits promising preventive and therapeutic effects on the rat model of TAA-induced hepatic encephalopathy, and these results provide substantial experimetal evidence for its clinical application.

BACKGROUNDTo observe cytopathogenic effect of Hantaan virus (HV) on cultured human bone marrow cells.

METHODSLight and transmission electron microscopy and direct immunofluorescent technique were applied to study cellular structure especially ultrastructural changes of bone marrow cells from patients with Hantaan virus infection. Bone marrow cells of one healthy volunteer were also studied as control.

RESULTSThe antigen of HV was found in bone marrow cells of 20 of 27 HFRS patients by the aid of direct immunofluorescent technique. It was found that the granulocytes had the highest percentage of HV antigen positive cells (76%), followed by monocytes (65%), lymphocytes (40%), megakaryocytes (20%) and the lowest was found in erythrocytes (3.7%). The injury of cell membrane after infection with HV was significantly more severe than that in the control group under the light and electron microscopy.

CONCLUSIONThis study demonstrated that HV could attack human bone marrow cells and cause cytopathogenic effect on them.

Adult ; Aged ; Antigens, Viral ; analysis ; Bone Marrow Cells ; ultrastructure ; virology ; Female ; Fluorescent Antibody Technique, Direct ; Hantavirus ; immunology ; pathogenicity ; Hemorrhagic Fever with Renal Syndrome ; pathology ; virology ; Humans ; Male ; Microscopy, Electron, Scanning ; Microscopy, Fluorescence ; Middle Aged

OBJECTIVETo develop a GC-FID method to determine borneol's concentration in mouse tissues, and to investigate the tissue distribution after intravenous and intranasal administrations of borneol.

METHODMouse brains, hearts, livers, spleens, lungs and kidneys were collected at 1, 3, 5, 10, 20, 30, 60, 90, 120 min after administration of borneol with the dose of 30.0 mg x kg(-1). The drug in tissues was extracted with ethyl acetate, and borneol's concentration detected by GC, with octadecane as the internal standard.

RESULTThe calibration curve showed a good linear relationship. Extraction recoveries, inter-day and intra-day precisions and stability were in conformity with the analytical requirements of biological samples. Borneol was mainly distributed in most tissues, more in heart, brain and kidney, and less in liver, spleen and lung.

CONCLUSIONThe established GC-FID method is applicable for content determination of borneol in tissues. After intravenous and intranasal administrations in mice, borneol is mainly distributed in abundant blood-supply tissues. After intranasal administration, brain tissues showed the highest target coefficient and target effectiveness.

Administration, Intranasal ; methods ; Administration, Intravenous ; methods ; Animals ; Bornanes ; administration & dosage ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Female ; Mice ; Mice, Inbred ICR ; Tissue Distribution

OBJECTIVETo observe the effect of Kangxin Capsule (KXC) on the expression of nerve growth factor (NGF) as well as the morphology and amount of nerve synapse in the cortical parietal lobe and hippocampus CA, area of vascular dementia ( VD) model rats.

METHODSThe model rats of VD made by photochemical reaction technique were randomly divided into five groups: the model group (MG), the high-dose, middle-dose and low-dose KXC groups (HDG, MDG and LDG), and the Western medicine hydergin control group (WMG). They were treated respectively with distilled water, high, middle and low dosage of KXC suspended liquid, and hydergin for a month. Besides, a blank group consisting of normal (non-model) rats was set up for control (CG). The ultrastructure of nerve synapse in the cortical parietal lobe and hippocampus CA1 area of the rats were observed and its density estimated. The condition of NGF positive neurons in the above-mentioned two regions were also observed by immunohistochemical stain.

RESULTSAll the KXC or hydergin treated groups demonstrated a normal amount of nerve synapse with integral structure in the cortical parietal lobe and hippocampus CA, area, which approached that in the CG and was superior to that in the MG. Also, the NGF positive neuron in all the treated groups was much more than that in MG with significant difference ( P<0.01), approaching to that in the CG.

CONCLUSIONKXC could elevate the expression of NGF in the cortical parietal lobe and hippocampus CA, area, preserve the number and morphology of synapse, thus to protect the function of nerve system from ischemic injury.

Animals ; Capsules ; Dementia, Vascular ; drug therapy ; metabolism ; pathology ; Disease Models, Animal ; Female ; Hippocampus ; chemistry ; Immunohistochemistry ; Male ; Medicine, Chinese Traditional ; Microscopy, Electron ; Nerve Growth Factors ; analysis ; Neurons ; ultrastructure ; Parietal Lobe ; chemistry ; Rats ; Rats, Wistar ; Synapses ; ultrastructure

To confirm the inactivating effect of chito-oligosaccharides on Tobacco mosaic virus (TMV) par ticles in vitro, the difference of TMV pathogenicity was evaluated according to the decrease of local lesion numbers after inoculating with TMV mixed with chito-oligosaccharides (DP3-10) in Nicotiana glutinosa, and the virion structural change was studied by transmission electron microscopy after mixed with chito-oligosaccharides. In the range of tested concentrations of chito-oligosaccharides (100-1000 microg /mL), the numbers of local lesions were strongly reduced with over 30% decrement, and the 88.4% reduction gained at the concentration of 600g /mL. It revealed that treatment with chito-oligosaccharide solution of 300-500 microg /mL directly broke TMV particles into tiny pieces of 50-150nm long, and that treatment with solutions of 600-1000 microg/mL caused virus particle agglomerated. The data presented here suggested that chito-oligosaccharides exerted strong inactivating effect on plant virus in vitro.
Microscopy, Electron, Scanning ; Oligosaccharides ; pharmacology ; Tobacco Mosaic Virus ; drug effects ; ultrastructure ; Virion ; drug effects ; ultrastructure

OBJECTIVE: To investigate the relationship between the mutation and abnormal expression of p16 gene in peripheral lung carcinoma and its CT manifestations. METHODS: Immunohistochemistry and PCR-SSCP were used to detect P16 protein expression and p16 gene mutation of 52 cases of peripheral lung cancer. All patients were scanned with spiral CT before the operation and proved by pathology. RESULTS: Of the 52 cases of lung cancer tissues, the negative expression rate of p16 gene protein was 53.8% (28/52), and the deletion or mutation rate of the exon 2 was 23.1% (12/52). There were significant statistical differences of p16 gene defect and its protein loss rates among groups of different clinical stages (P < 0.05), but among groups of different tissue types, different differentiation degree p16 gene defect and its protein loss rates showed no significant statistical difference (P > 0.05). In lung cancer patients with CT appearances of thin spicule, speculated protuberance, pleural indentation, and metastasis of lymph node, p16 gene and its protein loss rates were much higher than those without CT manifestations mentioned above (P < 0.05). However, there were no statistical differences among groups of different tumor sizes, with or without lobulation, with or without cavity, and different contrast enhanced CT values (P > 0.05). CONCLUSION p16 gene mutation and abnormal expression may play an important role in the occurrence and development of lung cancer, and it is relative to CT appearances of lung cancer. p16 gene may be used as a predicting index for clinical diagnosis and prognosis assessment.
Adult ; Aged ; Carcinoma, Squamous Cell ; diagnostic imaging ; genetics ; Female ; Gene Deletion ; Genes, p16 ; Humans ; Lung Neoplasms ; diagnostic imaging ; genetics ; Male ; Middle Aged ; Point Mutation ; Tomography, X-Ray Computed