Objective To observe the influence of tumor necrosis factor-alpha(TNFα) on skeletal muscle protein catabolism in rats with chronic obstructive pulmonary disease (COPD) and the effects of indomethacin (IND) on it. Methods Duplicated COPD model rats were divided into two groups: the malnutrition group and the normal nutrition group. The malnutrition group were further divided by randomized block design into four groups. Isotonic physiologic saline was administered to group A, the control and the normal nutrition group, and different doses of oral IND were administered to groups B, C, and D weight, concentrations of TNFα, contents of 3-methyl-histidine ( 3- M H ) and tyrosine (Tyr) in the diaphragm and extensor digitorum longus muscle homogenates were measured before and after the intervention. Results Before the intervention, the concentrations of TNFα in the serum of malnutrition groups were all significantly higher than those of normal nutrition group and the control group. After the intervention: (1) The concentrations of TNFα in the serum of the rats of group B, C and D were significantly lower than the group A, especially in group C. The levels of TNFα in serum and body weight of model group rats were negatively correlated ( r = -0. 846, P ＜0. 01 ), as well as the diaphragm and extensor digitorum longus muscle weights ( r = - 0. 778, P ＜ 0. 01; r = - 0. 772, P ＜ 0. 01 ). (2) The levels of 3-methyl-histidine in the diaphragm and extensor digitorum longus muscles of the intervention group C was lower than the COPD normal nutrition group, as well as the intervention groups B and D. The contents of tyrosine in the diaphragm and extensor digitorum longus muscles of the intervention group C was lower than that of the COPD normal nutrition group,as well as the groups B and D. The body weight growth value of the intervention group B were slightly higher than the group A, without significant difference( P ＞ 0. 05 ), while the group C was significantly higher than the group A ( P ＜ 0. 01 ). Conclusions TNFα is involved in the occurrence of COPD malnutrition and skeletal muscle amyotrophy. IND can reduce the TNFα levels in the serum and the catabolic rates of the skeletal muscle proteins in malnutrition rats with COPD, so as to improve partly the skeletal muscle atrophy.

This study was aimed to observe the effect ofZi-Bu Pi-Yin Recipe (ZBPYR) on the mRNA expressions of NMDA receptor subunits NR1, NR2A, NR2B in different brain regions of spleen-yin deficiency Alzheimer's Disease (AD) model rats. The levels of NR1, NR2A, NR2B mRNA expressions were detected by using RT-PCR method. The results showed that the levels of NR1, NR2A, NR2B mRNA expressions of AD group and spleen-yin deficiency AD group decreased significantly (P < 0.05). The levels of NR1, NR2A, NR2B mRNA expressions of ZBPYR treatment group increased significantly (P < 0.05). It was concluded that the expression levels of NMDAR mRNA in different brain regions of the ZBPYR treatment group increased significantly, which indicated that ZBPYR may up-regulate the protein expressions of NMDAR by increasing the expression levels of NMDAR mRNA, thereby to play the anti-dementia effect.

This study was aimed to observe the changes of dendritic spine density in different regions of brain among spleen-yindeficiency dementia (SYDD) model rats, in order to investigate the effects ofZi-Bu Pi-Yin Recipe (ZBPYR) on dendritic spines. Spleen-yindeficiency (SYD) rats were modeled by classical method. And incubatedβ-Amyloid 1-40 (Aβ1-40) was injected into the hippocampus of each rat to make SYDD model, which received the administration of ZBPYR. Golgi staining was used to stain dendritic spine in different regions of brain in rat model for the observation of the amount and shape. The results showed that dendritic spine density in different regions of hippocampus and cortex in SYDD group was reduced than that of the SYD group. Compared with the dementia group and the SYDD group, the dendritic spine density in different regions of hippocampus and cortex of the SYDD + ZBPYR group was increased. Compared with the blank control group, the dendritic spine density in different regions of hippocampus and cortex in rats from the dementia group was reduced. It was concluded that there were different degrees of reducing in the dendritic spine density of different brain regions in SYDD group. ZBPYR improved the learning and memory impairment, which might be related to the maintenance of dendritic spine density in different brain regions.

Objective To study the different ultrasonic features in patients of polycystic ovary syndrome (PCOS) with or without obesity based on body mass index (BMI), and to investigate whether certain hormonal factors correlate with ovarian morphology and blood flow, and to discuss the role of ultrasound combined with hormone test in the diagnosis of obese PCOS. Methods One hundred and five women with PCOS were recruited. Patients were divided into two groups according to BMI;obese PCOS group (OB-PCOS, n=32, BMI≥25 kg/m2) and non-obese PCOS (NOB-PCOS, n=73, BMI<25 kg/m2). The ultrasonic parameters of follicle number (FN), ovarian volume (Vol), resistance index (RI) of ovarian stromal blood, RI of uterine artery and serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), the ratio of luteinizing hormone and follicle-stimulating hormone (LH/FSH), progesterone (P), estradiol (E2), free testosterone (FT), prolactin (PRL), sex hormoe binding globulin (SHBG), fasting plasma glucose (FPG), fasting insulin (FINS), the extent of insulin resistance and hyperandrogenism (HOMA-IR) were measured and compared. The correlation of the ultrasonic parameters and hormonal factors were analyzed. Results The Vol of OB-PCOS group was significantly higher than NOB-PCOS group [(12.25±4.89) ml vs (10.73±2.30) ml, t=2.20, P < 0.05]. FN and uterine artery RI of OB-PCOS group had a rising trend and RI of ovarian interstitial was on a reducing trend compared with NOB-PCOS group. But the differences were not statistically significant. The levels of FINS and HOMA-IR in OB-PCOS group [(14.82±6.45) mU/L and (3.91±3.30)] were significantly higher than those in NOB-PCOS group [(8.04±4.57) mU/L and (1.64±1.20)] (t=4.87, 3.47, respectively, both P < 0.01). And FSH in NOB-PCOS group was significantly higher than OB-PCOS group [(5.95±1.91) U/L vs (4.65±1.88) U/L, t=-2.77, P<0.01]. In POCS patients, FN was significantly associated with LH/FSH (r=0.35, P<0.01), and FT (r=0.38, P<0.01). Vol was significantly associated with LH/FSH, BMI, HOMA-IR and FPG (r=0.27, P<0.05;r=0.25, P<0.05;r=0.40, P<0.01;r=0.32, P<0.01). RI of ovarian stromal blood flow was significantly associated with SHBG (r=0.28, P<0.05). In OB-POCS group, RI of uterine artery was significantly associated with PRL (r=-0.58, P < 0.05). Vol was significantly associated with HOMA-IR (r=0.47, P < 0.05). In NOB-POCS group, FN was significantly associated with LH/FSH (r=0.33, P<0.05), and FT (r=0. 56, P<0.05). Vol was significantly associated with FT (r=0.31, P < 0.05). Conclusion There are some differences in the ultrasound and endocrine parameters between obese and non-obese PCOS patients, and some correlations exist between them.

OBJECTIVETo observe the influence of heat shock preconditioning on the expressions of heat shock protein (HSP) 60 and HSP 70 and on the activities of cytochrome oxidase (CCO) and superoxide dismutase (SOD) in mitochondria in gastric mucosa of severely scalded rats, and to investigate its protective mechanism on acute gastric mucosal lesion in rats with severe scald.

METHODSOne hundred and forty-four Wistar rats were randomized into three groups, i. e. scald group ( n = 40, acute gastric mucosal lesion was made after scald, other 8 normal rats without scald were employed as blank control); HS group ( n =40, with heat shock preconditioning 20 h before scald), and other 8 rats preconditioned with heat shock but without scald were employed as experimental control I; actinomycin D group ( n = 40, with intraperitoneal injection of 0. 1 mg/kg actinomycin D 30 min before heat shock preconditioning and other treatment as HS group), and other 8 rats with merely actinomycin D injection were employed as experimental control II. Eight rats in each group were sacrificed and laparotomized at 3, 6, 12, 24 and 48 post-scald hours (PSH) , respectively to determine the index of gastric mucosal lesions (UI ) , the mRNA expressions of HSP70 and protein expression of HSP60 and HSP70, and the changes in the activities of SOD and CCO.

RESULTSUI of the scalded rats increased as the time elapses, reaching the peak (12. 8 +/- 1.9) at 24 PSH. In addition, UI in HS group was significantly lower than that in scald group at each time-point except that at 3 PSH ( P < 0. 05 or 0. 01). The extent of gastric mucosal lesion in rats in actinomycin D group was obviously aggravated compared with that in scald and HS groups ( P <0. 05). The HSP70 mRNA expression in both scald and HS groups was increased at each time-points except for 48PBH, while that in actinomycin D group was increased at 24 PBH and 48PBH. The expressions of HSP70 and HSP60 were greatly increased in HS group compared with those in scald group ( P < 0. 05 or 0. 01) , while those in actinomycin D group were significantly inhibited ( P < 0. 05). The activities of CCO and SOD were gradually decreased in gastric mucosa in scald group, but it was greatly improved by HS preconditioning at 6, 12, 24 PSH ( P < 0. 05 or 0. 01).

CONCLUSIONHeat shock preconditioning is beneficial for the protection of acute gastric mucosal lesion of rats after severe scald, due to increase of HPS60 and HSP70 expression, and increase of CCO and SOD activities in mitochondria.

Animals ; Burns ; metabolism ; pathology ; Chaperonin 60 ; metabolism ; Electron Transport Complex IV ; metabolism ; Female ; Gastric Mucosa ; metabolism ; pathology ; HSP70 Heat-Shock Proteins ; metabolism ; Heat-Shock Response ; Male ; Mitochondrial Proteins ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism

Adult ; Anemia, Aplastic ; chemically induced ; genetics ; immunology ; Benzene ; poisoning ; Gene Expression ; Genes, T-Cell Receptor beta ; genetics ; Humans ; Male

Objective To investigate the usefulness of 99Tcm-MDP whole body bone scintigraphy (WBBS) in patients with synovitis,acne,pustulosis,hyperostosis,osteitis (SAPHO) syndrome.Methods 99Tcm- MDP WBBS was performed in 25 patients (6 males,19 females,mean age =(55.1 ±9.8) years)with SAPHO syndrome.Bone lesions were classified into five categories:anterior chest wall,spine,mandible,sacroiliac joint,and limbs.The typical scintigraphic manifestations of SAPHO syndrome were summarized and compared to other radiological imaging data.Results Among 25 patients,32％ of cases (8/25)were associated with skin lesion; 48％ ( 12/25 ) were pathologically diagnosed with chronic nonspecific bone inflammation by bone biopsy.On 99Tcm-MDP WBBS,abnormal metabolic foci at anterior chest wall were found in all cases,most of which located in the sternocostoclavicular region (96％,24/25 ),including sternoclavicular joints (60％,15/25),first costosternal junctions (48％,12/25),and manubriosternal junctions (44％,11/25 ).Only 20％ of the patients (5/25) demonstrated the typical scintigraphic characteristic:“bull's head” sign.The second most frequent part was spine (44％,11/25).Appendicular skeleton was affected in 16％ (4/25) patients.WBBS also demonstrated additional skeletal lesions in 68％ (17/25 ) of the patients,mainly in first costosternal junctions (7 patients),sternoclavicular joints (6 patients),manubriosternal junctions (5 patients) and spine (4 patients).Conclusions Abnormal metabolic foci in sternocostoclavicular region and other imaging manifestations on 99Tcm- MDP WBBS can be used to diagnose,differentiate,and localize the insidious lesion and evaluate the lesion activity in patients with SAPHO syndrome.

Objective To investigate the association of angiotensinogen(AGT)gene A-6G、T174M and G-217A polymorphisms with the risk of essential hypertension(EH)in the elderly of Han nationality.Methods Genotypes of AGT gene A-6G,T174M and G-217A polymorphisms in 177 aged EH patients and 86 sex and age-matched controls were analyzed with gene chip technology.Results The A-6G and T174M polymorphisms of AGT gene were significantly associated with EH.The numbers of the three genotypes of A-6G were 113,58 and 6 in the patient group and 70,15 and 1 in the control group(P= 0.014)and those of T174M were 94,77 and 6,60,25 and 1(P=0.031),respectively.G-217A polymorphism was not related to EH.Individuals carrying A-6G AA and T174M CC genotypes showed 57% and 56% lower risk of EH(OR=0.43;95%CI=0.23-0.82 and OR=0.44;95%CI=0.25-0.79, respectively).Conclusions The A-6G AA and the T174M CC genotype may be related with decreased risk of EH and G-217A polymorphism may have little role in the etiology of EH in Han nationality.

BACKGROUNDNF-kappaB p65 was shown to inhibit transcription of phosphoenolpyruvate carboxykinase (PEPCK), a rate-limiting enzyme in gluconeogenesis in the liver. To understand the mechanism of action of NF-kappaB p65, we investigated the nuclear receptor corepressor in the regulation of PEPCK transcription.

METHODSRat H4IIE cells, human hepatoma HepG2 cells and human embryo kidney (HEK) 293 cells were used in this study. The transcriptional activity of a rat PEPCK gene promoter (-490/+100) was analyzed in HepG2 cells, a HepG2 super suppressor IkBalpha (ssIkBalpha) stable cell line, and HEK 293 cells. The effects of p65 and ssIkBalpha on a rat PEPCK gene promoter were observed using the PEPCK luciferase reporter system. The interaction of the cAMP-response- element-binding (CREB) protein, histone deacetylase 3 (HDAC3) and silencing mediator for retinoic and thyroid hormone receptors (SMRT) with the PEPCK gene promoter were investigated using the chromatin immunoprecipitation (ChIP) assay. p65 cotransfection and RNAi-mediated gene knockdown were used to determine the corepressor involved in the inhibition of PEPCK by NF-kappaB p65 and the transcriptional regulation of CREB by NF-kappaB p65.

RESULTSNF-kappaB p65 inhibited PEPCK expression and the inhibition was blocked by ssIkBalpha. The inhibitory effect of p65 was completely blocked in a HepG2 stable cell line in which ssIkBalpha was expressed. HDAC3 or SMRT knockdown led to a significant up-regulation of PEPCK reporter activity in the presence of p65 cotransfection. In the ChIP assay the interaction of HDAC3 and SMRT with the PEPCK gene promoter was induced by p65 activation, but the CREB signal was reduced. Transcriptional activity of CREB was inhibited by NF-kappaB p65 cotransfection. The inhibitory effect of NF-kappaB p65 was blocked by HDAC3 RNAi or SMRT RNAi.

CONCLUSIONSThe study showed that the inhibition of PEPCK by NF-kappaB p65 was dependent on HDAC3 and SMRT, which form a nuclear corepressor complex for transcriptional inhibition. The transcription factors NF-kappaB p65 and CREB share the same corepressor HDAC3-SMRT, and the corepressor exchange leads to inhibition of PEPCK gene transcription by NF-kappaB p65.

Animals ; Blotting, Western ; Cell Line ; Chromatin Immunoprecipitation ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Hep G2 Cells ; Histone Deacetylases ; genetics ; metabolism ; Humans ; NF-kappa B ; genetics ; metabolism ; Nuclear Receptor Co-Repressor 2 ; genetics ; metabolism ; Phosphoenolpyruvate Carboxykinase (ATP) ; genetics ; Promoter Regions, Genetic ; genetics ; Protein Binding ; genetics ; physiology ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factor RelA ; genetics ; metabolism

OBJECTIVETo assess the feasibility of using MTT assay for detecting the apoptosis of osteosarcoma cells following chemotherapy.

METHODSThe osteosarcoma cells derived form surgical specimens were cultured in RPMI 1640 medium supplemented with 10% calf serum. Chemotherapeutic agents were administered in the cell culture, and MTT assay was used to observe the cell apoptosis under optical microscope.

RESULTSMTT staining accurately identified apoptosis of the osteosarcoma cells, and the apoptotic cells were easily distinguished from normal cells and dead cells.

CONCLUSIONMTT staining is convenient and practical for detecting osteosarcoma cell apoptosis.

Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Apoptosis ; drug effects ; Bone Neoplasms ; pathology ; surgery ; Humans ; Osteosarcoma ; pathology ; surgery ; Tetrazolium Salts ; Thiazoles ; Tumor Cells, Cultured