Objective The aim of this study was to explore the effect of androgen deficiency on serum hormone levels, visceral fat accumulation and inflammatory gene expression in miniature pigs fed a high-fat diet ( HFD) . Methods Sexually mature male Chinese Wuzhishan miniature pigs were divided into three groups ( animals/group) as follows:intact male pigs ( SHAM) , castrated male pigs ( CAS) and castrated male pigs plus testosterone treatment ( CAS+T) . The pigs were fed a HFD diet for 12 weeks. Serum levels of testosterone and leptin were measured and visceral fat were dissected and weighted. qRT?PCR was performed to determine the mRNA expression levels of lipogenic, lipolysis and inflammation relat?ed genes. Results (1) Serum testosterone levels were significantly decreased but serum leptin levels were significantly in?creased in the castrated pigs. These effects were recovered after testosterone treatment. ( 2 ) Visceral fat percentage was significantly increased in the castrated pigs, and testosterone treatment reduced the increased visceral fat in the castrated pigs. (3) Castration and testosterone treatment had no significant effects on the expression levels of lipogenic genes (FAS and ACC) and lipolysis genes (HSL and ATGL) in pigs fed a HFD. (5) Castration significantly induced the expressions of inflammatory genes including Leptin, CD68, CCL16, CCL23 and SAA, and testosterone treatment recovered the expres?sions of the above genes in the castrated pigs. Conclusions Castration?induced testosterone deficiency promotes visceral fat accumulation and upregulates the expression levels of inflammatory genes in miniature pigs fed a HFD. Moreover, tes?tosterone treatment ameliorates castration?induced visceral fat accumulation and inflammatory response in HFD?fed pigs.

The binding of pefloxacin mesylate (PFLX) to bovine lactoferrin (BLf) and human serum albumin (HSA) in dilute aqueous solution was studied using fluorescence spectra and absorbance spectra. The binding constant K and the binding sites n were obtained by fluorescence quenching method. The binding distance r and energy-transfer efficiency E between pefloxacin mesylate and bovine lactoferrin as well as human serum albumin were also obtained according to the mechanism of Förster-type dipole-dipole nonradiative energy-transfer. The effects of pefloxacin mesylate on the conformations of bovine lactoferrin and human serum albumin were also analyzed using synchronous fluorescence spectroscopy.
Animals ; Anti-Bacterial Agents ; chemistry ; metabolism ; pharmacology ; Binding Sites ; Cattle ; Humans ; Kinetics ; Lactoferrin ; chemistry ; metabolism ; Pefloxacin ; chemistry ; metabolism ; pharmacology ; Protein Binding ; Protein Conformation ; Serum Albumin ; chemistry ; metabolism ; Spectrometry, Fluorescence ; Spectrophotometry, Ultraviolet

Aim To investigate the effect of aloe-emodin on lipopolysaccharide(LPS)-induced production of nitric oxide and expression of inducible nitric oxide synthase in RAW264.7 cells.Methods RAW264.7 macrophage line in mice was induced by LPS to set up the inflammatory model.Nitric oxide(NO)production was examined by Griess reaction;the expression of iNOS mRNA was detected by RT-PCR analysis;NO radical generation was tested by sodium nitroprusside method.Results Aloe-emodin at the dose of 0.69～2.5 mg·L~(-1) exhibited the inhibitory effect on LPS-induced NO production in a dose-dependent and time-dependent manner;aloe-emodin at the dose of 0.63～5.00 mg·L~(-1) suppressed LPS-induced iNOS mRNA expression in RAW 264.7 cells.However,aloe-emodin had no scavenging effect on sodium nitroprusside-triggered NO production,and didn't affect iNOS enzyme activity.Conclusion Aloe-emodin inhibited signifi-cantly LPS-induced NO production through suppressing inducible NO synthase(iNOS)expression at mRNA level in a dose-dependent and time-dependent manner,but failed to affect sodium nitroprusside-triggered NO production and iNOS enzyme activity.

Head ; surgery ; Humans ; Neck ; surgery ; United States

The data mining technology was introduced, and the law and peculiarity of TCM syndrome and treatment were analyzed in this paper. Concurrently, the application of the data mining in TCM researches on rule of syndrome and treatment was roughly summarized and its advantages and existing problems were pointed out. In order to enhance the efficiency and the appliance value of the mining technology and to accelerate the development of TCM research, the method for integrating multi-algorithms to make the mining was proposed.
Biomedical Research ; Data Mining ; methods ; Diagnosis, Differential ; Drug Therapy ; Humans ; Medicine, Chinese Traditional

OBJECTIVETo investigate the protective effect of phosphodiesterase type 5 inhibitors (tadalafil) on the testis following testicular ischemia-reperfusion injury in rats.

METHODSEighty-four healthy adult male SD rats were randomly and equally divided into groups A (sham operation), B (testicular torsion + low-dose tadalafil), C (testicular torsion + high-dose tadalafil), and D (testicular torsion + placebo). Models were established in the latter three groups by 7200 torsion of the right testis for 2 hours. The animals in groups A and B were treated by gavage with tadalafil at the dose of 0. 5 mg per kg per day, those in group C at 2 mg per kg per day, and those in group D with saline at the same dose. After 3, 7, and 14 days of treatment, the torsioned testes were harvested for evaluation of the superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the testis tissue. The pathological changes in the testis were observed under the light microscope.

RESULTSAt 3, 7, and 14 days, the SOD activity was (254.46 +/- 7.43), (278.49 +/- 8.33), and (317.99 +/- 3.31) nU/mg prot in group B, and (277.12 +/- 8.80), (309.40 +/- 2.14), and (320.39 +/- 4.72) nU/mg prot in group C, all obviously higher than in D ([223.21 +/- 4.65], [231.45 +/- 4.16] and [248.28 +/- 5.74] nU/mg prot), while the MDA content was lower in the former two groups than in the latter. At 3 and 7 days, the SOD activity was significantly higher and the MDA level significantly lower in group C than in B (both P < 0.01) , while at 14 days, neither showed any remarkable differences between the two groups (P > 0.05). No obvious histopathological change was observed in the testis tissue of group A. At 3 and 7 days, pathological examination of the testis tissue revealed significant differences in the number of seminiferous epithelial layers, testicular histological score, and seminiferous tubule diameter in group B (P < 0.01), but the three indexes at 14 days in group B and at 7 days in group C exhibited no remarkable differences from those at 14 days in group A.

CONCLUSIONTadalafil can alleviate testicular ischemia-reperfusion injury following testis torsion/detorsion in a time- and dose-dependent manner.

Animals ; Biomarkers ; metabolism ; Carbolines ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Male ; Malondialdehyde ; metabolism ; Phosphodiesterase 5 Inhibitors ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Seminiferous Tubules ; pathology ; Spermatic Cord Torsion ; complications ; Superoxide Dismutase ; metabolism ; Tadalafil ; Testis ; blood supply ; metabolism ; pathology ; Time Factors

In the experiment, we designed and synthesized two siRNAs based on the sequence of nuclear receptor-related factor 1 (Nurr1) mRNA. They were separately subcloned into the plasmid of pSilenCircle (pSC) containing U6 promoter. The pSC-Nurr1 vectors (pSC-N1 and pSC-N2) specific to Nurr1 gene and the negative control vector of short-hairpin RNA (shRNA) eukaryotic expression vector were constructed. We cultured the dopaminergic cell line MN9D and the verified vectors were transfected with LipofectamineTM 2000 in vitro. The positive cell clones transfected with pSC were obtained after being screened with 500 mug/ml G418. After that, the silencing effects of Nurr1 and TH mRNA or protein were detected by real time RT-PCR and Western blot. The neurite extension of MN9D cells was observed and photographed by inverted microscope. The results showed that Nurr1 mRNA expression in MN9D cells was specifically down-regulated by the vectors of pSC-N1 and pSC-N2, and the silencing effects were 62.3% and 45.6%, respectively. The dopaminergic phenotype of TH mRNA was also suppressed significantly and the silencing effects were 76.3% and 62.6%, respectively. Meanwhile, the expressions of Nurr1 and TH proteins were also significantly suppressed, and the silencing effects of Nurr1 and TH protein were 57.4%, 72.0% and 79.1%, 70.1% respectively. The negative control and liposome groups had no effect on the two genes. In conclusion, Nurr1 shRNA expressing vectors can inhibit the expressions of Nurr1 and TH mRNA or protein in MN9D cells, and Nurr1 might play a role in neurite extension of MN9D cells. Nurr1 shRNA expressing vector may provide a novel applicable strategy for the study on the function of the genes associated with Parkinson disease and the development of dopaminergic neuron.
Cell Line ; Dopaminergic Neurons ; cytology ; metabolism ; Down-Regulation ; Fetus ; Humans ; Mesencephalon ; cytology ; Neurites ; physiology ; Nuclear Receptor Subfamily 4, Group A, Member 2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Tyrosine 3-Monooxygenase ; genetics ; metabolism

OBJECTIVETo observe clinical curative effect of cake-separated moxibustion on impaired glucose regulation (IGR) and explore its action mechanism.

METHODSSixty cases were randomly divided into a simple lifestyle intervention group (control group) and a cake-separated moxibustion combined with lifestyle intervention group (observation group), 30 cases in each one. The control group was treated with lifestyle intervention. Based on lifestyle intervention, cake-separated moxibustion at Pishu (BL 20), Weishu (BL 21) and Yishu (EX-B 3) was applied to the observation group. Fast plasma glucose (FPG), two hours plasma glucose after oral glucose tolerance test (OGTT2hPG), fasting insulin (FINS), homa insulin resistance index (HOMA-IR), blood lipid, body mass index (BMI) and waist circumference (WC) were observed in the two groups before and after treatment.

RESULTSAfter treatment, the OGTT2hPG and FPG were both decreased significantly (both P<0.05) in the two groups, compared between the two groups, the differences of FPG [(0.41 +/- 0.42) mmol/L vs (0.05 +/- 0.08)mmol/L] and OGTT2hPG [(0.85 +/- 0.53)mmol/L vs (0.17 +/- 0.19)mmol/L] were both statistically significant. There were no significant changes in FINS, HOMA-IR, blood lipid, BMI and WC in the control group before and after treatment (all P>0.05), but FINS, HOMA-IR levels, triglycerides (TG), total cholest-erol (TC), low density lipoprotein (LDL-C), BMI and WC in the observation group were decreased obviously after treatment (all P<0.05), which had statistical differences between the two groups (all P<0.05).

CONCLUSIONThe cake-separated moxibustion combined with lifestyle intervention can obviously control blood glucose levels, improve insulin resistance and blood lipid levels, decrease BMI and WC.

Adult ; Aged ; Female ; Glucose ; metabolism ; Glucose Intolerance ; metabolism ; physiopathology ; therapy ; Humans ; Insulin ; Male ; Middle Aged ; Moxibustion ; Waist Circumference

OBJECTIVETo investigate apoptosis-inducing effect and its mechanisms of HY-1, a carbazole alkaloid, on human erythroleukemia K562 cells.

METHODSCell proliferation was detected by sulforhodamine B (SRB) assay after treated with HY-1 at indicated doses. Cell cycle analysis was performed by flow cytometry, mitochondria membrane voltage change was assessed by rhodamine 123 staining, annexin V-PI apoptosis detecting kit and DNA agarose gel electrophoresis were used to identify apoptosis-inducing effect of HY-1. The alterations of apoptosis-relating proteins were detected by Western blot.

RESULTSThe IC(50) of HY-1 in K562 cells was (29.05 +/- 0.90) micromol/L by SRB assay. HY-1 had significant apoptotic inducing effect on K562 cells in a dose- and time-dependent manner as verified by appearance of Sub-G(1) peak on histogram of flow cytometry analysis, reduction of mitochondria membrane voltage, appearance of double positive cell group in Annexin V-PI apoptosis detecting test, and remarkable DNA ladder. The expression of cytosolic cytochrome c was apparently increased. Pro-caspase-9, pro-caspase-3 and PARP were all cleaved to active segments. There was no change in the expression of caspase-8.

CONCLUSIONHY-1 exerts its anticancer activity through triggering apoptosis of K562 cells by mitochondria-activating pathways.

Alkaloids ; isolation & purification ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Carbazoles ; isolation & purification ; pharmacology ; Humans ; K562 Cells ; Mitochondria ; metabolism ; Rutaceae ; chemistry

The study was aimed to explore the trehalose method for storing platelets in cold. (51)Cr-labeling platelet was used to detect the platelet survival. The platelet function in vitro was performed by platelet aggregate analyzer. After treatment with 50 mg/ml trehalose at 37 degrees C for 4 hours, the rabbit platelet concentrates (PC, 2.0 x 10(9)/ml) were stored in 4-8 degrees C refrigeration, the platelet function in vitro and survival of chilled platelets transfused into self-rabbits were observed. The results showed that trehalose could protect the chilled rabbit platelets. After PC stored at 20-24 degrees C and 4-8 degrees C for up to 24 hours, the platelet aggregate in vitro in response to 11.2 micromol/L ADP were (75.3 +/- 9.8)% and (80.5 +/- 12.5)%, the survival of PC stored at 20-24 degrees C and 4-8 degrees C for 24, 48, 72 hours after transfused into self-rabbits were (78.1 +/- 7.9)%, (65.4 +/- 6.7)%, (57.5 +/- 7.2)% and (5.1 +/- 2.5)%, (2.8 +/- 2.0)%, (0.9 +/- 0.8)%, respectively. The PC treated with 50 mg/ml trehalose were remained stable for up to 12 days of refrigerated storage in autologous plasma. The platelet aggregate in vitro in response to 11.2 micromol/L ADP at 12 days after stored in refrigeration was (77.8 +/- 9.5)%, the survival on 24, 48, 72 hours of platelet transfused into self-rabbits were (75.7 +/- 11.0)%, (67.0 +/- 8.5)%, (56.8 +/- 8.0)%, respectively. Compared with control group of storing at 20-24 degrees C for 24 h, P > 0.05. In conclusion, trehalose can protect the chilled blood platelets, prolong the circulation of refrigerated rabbit platelets, and not impair chilled rabbit platelet function.
Adenosine Diphosphate ; pharmacology ; Animals ; Blood Platelets ; cytology ; drug effects ; Blood Preservation ; methods ; Cell Survival ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Platelet Aggregation ; drug effects ; Platelet Transfusion ; Rabbits ; Trehalose ; pharmacology