Objective To investigate the effects of penehyclidine hydrochloride on activities of nuclear factor kappa B (NF-kB) and activator protein-1 (AP-1) during actue lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation (HSR) in rats.Methods Thirty male Sprague-Dawley rats,aged 8 weeks,weighing 250-300 g,were randomly assigned into 3 equal groups (n =10 each) using a random number table:sham operation group (group S),blunt chest trauma-HSR group (group THSR) and penehyclidine hydrochloride group (group PHCD).The model of actue lung injury induced by blunt chest trauma-HSR was induced by dropping a 300 g weight onto a precordium in anesthetized rats.Blood was withdrawn via the femoral artery 5 min later until MAP was decreased to 35-45 mmHg within 15 min and maintained at this level for 60 min,followed by resuscitation.In PHCD group,PHCD 2 mg/kg was injected intravenously at 60 min after hemorrhagic shock.At 6 h after the model was established,blood samples were obtained for measurement of concentrations of tumor necrosis factor-alpha (TNF-α) in serum.The lungs were then removed for determination of lung water content,myeloperoxidase (MPO) activaty (by colorimetric assay),NF-κB and AP-1 activaties (using electrophoretic mobility shift assay) in lung tissues,and for microscopic examination of pathologic changes (under light microscope).The left lung was lavaged,and lung permeability index (LPI) was calculated.Results Compared with S group,lung water content,LPI,serum TNF-α level and activites of MPO,NF-κB and AP-1 were significantly increased in THSR and PHCD groups.Compared with THSR group,lung water content,LPI,serum TNF-α concentrations and activites of MPO,NF-κB and AP-1 were significantly decreased in PHCD group.The pathological damage to lung tissues was significantly reduced in PHCD group as compared with THSR group.Conclusion PHCD can inhibit activities of NF-κB and AP-1 in lung tissues,thus mitigating acute lung injury induced by blunt chest trauma-HSR in rats.

Objective To study the effects of chrysophanol ( Chry ) on ammonia-induced cell swelling and gultamate uptake inhibition in cul-tured astrocytes and its signaling pathways .Methods Primary cultured mouse astrocytes were divided into normal group , model group , three Chry test group ( 0.1 , 1.0 , 10.0 μg · mL-1 ) and six positive control group[L-NAME 250 μmol· L-1, vitamin E 100 μmol· L-1, supero-xide dismutase ( SOD ) 25 U · mL-1 , extracellular regulated kinase 1/2 ( ERK1/2 ) inhibitor UO126 10 μmol · L-1 , P38MAPK inhibitor SB239063 10 μmol · L-1 , JNK1/2/3 inhibitor SP600125 1 μmol· L-1 ].Cultured astrocytes treated for 48 h, cell volume were measured by measuring the volume of water in cell and gultamate uptake were measured by the isotope labeling method after 72 h.The glutamate/aspartate transporter ( GLAST ) mRNA expression was detected by RT-PCR.Results Ammonia-induced astrocytes swelling and gulta-mate uptake inhibition and GLAST mRNA expression downregulation were reduced by moderate and lager doses Chry , which similar to the positive control.Conclusion Chry may improve the ammonia-induced astrocytes dysfunction by againsting oxidative stress and inhibiting theMAPKs phosphorylation.

AIMTo investigate the mechanisms of anti-cancer effect of resveratrol (Res), and the effects of Res in cell apoptosis. The role of Res playing in mitochondrial permeability transition pore (PTP) induction was studied.

METHODSMitochondria was prepared from the liver of Wistar rats. The effects of Res on oxygen consumption of isolated mitochondria from rat liver was measured with Clark-type electrode and resulted in respiration control rate (RCR). Mitochondrial swelling affected by Res was assessed spectrophotometrically, through the changes in absorbance at 540 nm. The PTP opening was learned from the results. Membrane potential of mitochondia was measured through fluorescence spectrophotometry.

RESULTSRes was shown to inhibit the respiration and decrease the RCR of mitochondria. Res can promote the PTP opening mediated by Ca2+. Res was shown to promote the increase of mitochondial membrane potential mediated by Ca2+ and loss of mitochondial membrane potential.

CONCLUSIONRes was shown to inhibit mitochondial respiration and induce PTP opening of mitochondria. These may be one of the pathways that Res showed anti-cancer action and induce cells apoptosis.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Calcium ; metabolism ; Female ; Ion Channels ; drug effects ; metabolism ; Membrane Potentials ; drug effects ; Mitochondria, Liver ; drug effects ; physiology ; Mitochondrial Membrane Transport Proteins ; Mitochondrial Swelling ; drug effects ; Rats ; Rats, Wistar ; Stilbenes ; pharmacology

Objective To discuss a real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) wether if can be used to detect Brucella. Methods According to the BCSP31 gene sequences specific for Brucella, one pair of primers and one TaqMan probe were designed. A real-time PCR was developed with the BCSP31 fragments cloned into PMD18-T vector. The standard cure was established and the sensitivity, the species specificity and the stability of the assay were evaluated. The clinical blood specimens were detected by QT-PCR and compared with clinical diagnosis. Results The standard curve was established with the standard template and the relationship between the Ct and the DNA copy number was linear(r=0.999). The sensitivity of the real-time PCR was 5 copies/μl. The sensitivity of the common PCR was 5×102 copies/μl. The sensitivity was about 100 times higher than common PCR. Species specificity of this FQ-PCR assay evaluated using genomic DNA from 6 Bmcella strains and 5 non-Brucella strains and strong fluorescence was detected in all Brucella strains. The CV of intra-assay and inter-assay reproducibility were 0.71%,7.23%, reprectively. Twenty-four specimens from clinical brucellosis cases, 19 showed positive, the positive coincident rate was 79%(19/24). The negative results were obtained for all 31 negative control, and the negative coincident rate was 100%(31/31). Two were positive from all 30 specimens clinically suspected. Conclusions Highly specific, sensitive, repeatable and coincidental with clinic, this FQ-PCR is quite useful for rapid detection of tiny DNA of Brucella in various samples and laboratory diagnosis.

OBJECTIVETo detect serve acute respiratory syndrome-associated coronavirus (SARS-CoV) and SARS-like-CoV in fruit bats captured in Guangzhou and its vicinity.

METHODSTotally 927 bats of 9 species (Cynopterus sphinx, Rousettus leschenaulti, Miniopterus schreibersi, Hipposideros pratti, Rhinolophusasinicus, Scotophilusakuhlii, Hipposideros Pomona, Rhinolophus affinis, and Rhinolophus pusillus) captured in Guangzhou and its vicinity from September 2004 to November 2005 were available for this investigation, from which 3,043 samples (813 throat swasb, 524 sera, 853 lung tissues and 853 colorectal tissue specimens) were obtained. SARS-Cov and SARS-like-CoV were detected in these specimens using diagnostic kit for novel coronavirus N protein (ELISA), SARS-CoV Virus RNA detection kit, fluorescence PCR, Genchip, RT-PCR and cell isolation culture methods.

RESULTS AND CONCLUSIONNo SARS-CoV and SARS-like-CoV were detected in the 3043 samples, indicating the current absence of SARS-CoV and SARS-like-CoV in the bats captured in Guangzhou and its vicinity.

Animals ; China ; epidemiology ; Chiroptera ; virology ; Disease Vectors ; Enzyme-Linked Immunosorbent Assay ; Humans ; Nucleocapsid Proteins ; metabolism ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; metabolism ; Severe Acute Respiratory Syndrome ; epidemiology ; transmission ; virology

OBJECTIVETo investigate the protective effect of L-carnitine (LC) combined with sildenafil on the reproductive endocrine function of male rats with diabetes mellitus (DM).

METHODSA total of 40 male SD rats were randomly divided into five groups, group A taken as normal controls, and groups B, C, D and E made into DM models by injection of streptozotocin at 65 mg/kg. Then the rats in groups A and B were treated with normal saline, C with sildenafil at 5 mg per kg per d, D with LC at 300 mg per kg per d, and E with sildenafil at 5 mg per kg per d plus LC at 300 mg per kg per d, all via gastric gavage for 6 weeks, followed by determination of the levels of testosterone (T), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the serum of the rats.

RESULTSAfter 6 weeks of treatment, the T, FSH and LH levels were (25.25 +/- 2.67) nmol/L, (5.78 +/- 0.61) IU/L and (625.21 +/- 43.45) ng/L in group A, (9.63 +/- 1.71) nmol/L, (1.98 +/- 0.42) IU/L and (479.89 +/- 27.62) ng/L in group B, (18.98 +/- 3.07) nmol/L, (5.08 +/- 0.33) IU/L and (586.57 +/- 31.72) ng/L in group C, (16.18 +/- 2.65) nmol/L, (4.63 +/- 0.30) IU/L and (540.78 +/- 25.52) ng/L in group D, and (23.65 +/- 2.66) nmol/L, (5.59 +/- 0.48) IU/L and (621.53 +/- 36. 40) ng/L in group E. The three parameters were significantly lower in B than in the other four groups (P < 0.01), and so were they in C and D than in A and E (P < 0.05), but showed no significant differences either between C and D (P > 0. 05) or between A and E (P > 0.05).

CONCLUSIONSix-week medication of either sildenafil or LC alone could increase the levels of T, FSH and LH in the serum of DM rats, but the combination of the two had an even more obvious increasing effect, which indicates a still better protective effect on the reproductive endocrine function of diabetic male rats.

Animals ; Carnitine ; adverse effects ; therapeutic use ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Drug Therapy, Combination ; Follicle Stimulating Hormone ; blood ; Luteinizing Hormone ; blood ; Male ; Piperazines ; administration & dosage ; therapeutic use ; Purines ; administration & dosage ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Sildenafil Citrate ; Sulfones ; administration & dosage ; therapeutic use ; Testosterone ; blood

OBJECTIVETo evaluate the precision of rat bone mineral density (BMD) measurements by Norland Excellplus dual-energy X-ray absorptiometry (DXA) and to investigate the BMD changes in ovariectomized (Ovx) rats in vitro.

METHODS(1) The coefficients of variation (CV) for BMD measurements at various skeletal regions were repeatedly determined by DXA in 10 Wistar rats in vitro. (2) BMD in lumbar vertebra (L5) and both sides of femurs was measured in total 90 rats. And (3) changes in BMD between Ovx and sham rats were compared.

RESULTS(1) The short-term CVs of BMD measurements in different regions by DXA were as follows, 1.58% for lumbar vertebra (L5), 0.90% for left femur, and 0.86% for right femur, respectively. The long-term CVs were 2.22% for lumbar vertebra (L5), 1.09% for left femur, and 1.20% for right femur. (2) The BMD values in 90 Wistar rats were (127.5 +/- 12.3) in lumbar vertebra (L5), (82.6 +/- 11.3) in corpus vertebra (L5'), (150.7 +/- 10.6) in left femur and (149.9 +/- 10.6) mg/cm2 in right femur, respectively. The correlation coefficient of BMD measurements between left and right femurs was 0.792 (P < 0.001). (3) In Ovx group, the BMD values of corpus vertebra (L5') and distal femurs were significantly decreased, that was 10.0%-17.5% lower in comparison with sham group (P < 0.001).

CONCLUSIONSMeasurement of rat BMD in vitro by Norland Excellplus DXA is a useful method, and it can reflect the changes in rat bone masses with good precision.

Absorptiometry, Photon ; Animals ; Bone Density ; Female ; In Vitro Techniques ; Ovariectomy ; Rats ; Rats, Wistar

OBJECTIVETo ascertain whether other variations coexist with 1555(A--> G) mutation in the mitochondrial DNA and may aggravate the severity of hearing loss or increase the penetrance of 1555(A--> G) mutation in a large family with maternally inherited nonsyndromic deafness in Huaiyin, Jiangsu province.

METHODSPCR-restriction fragment length polymorphism (PCR-RFLP) was used to screen both the nt1555 and the nt7445 of the mitochondrial DNA from 27 maternal members in the core family; and then the mitochondrial genomes from two maternal members, and the 12S rRNA genes MTRNR1 and tRNA-Ser(UCN) gene MTTS1 from the others, were amplified by PCR-RFLP and were sequenced.

RESULTS1555(A--> G) mutation in the mitochondrial DNA was reverified to be one of the major factors which cause maternally inherited nonsyndromic deafness and the cosegregation of 955-960(insC) and 1555(A--> G) was present in this family. Moreover, 7449 (insG), a novel homoplasmic mutation in the tRNA-Ser(UCN) gene, was found to co-exist with 1555(A--> G) mutation in two maternal members.

CONCLUSIONThe cosegregation of 955-960(insC) and 1555(A--> G) implies that 955-960(insC) may synergistically cause hearing loss in the presence of an 1555(A--> G) mutation, serving as an aggravating factor to enhance the sensitivity to aminoglycosides, and may sometimes increase the penetrance of 1555(A--> G) mutation.

DNA, Mitochondrial ; chemistry ; genetics ; Deafness ; genetics ; Female ; Genetic Predisposition to Disease ; Genome, Mitochondrial ; genetics ; Humans ; Male ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA

Objective To study the genetic characterization of enterovirus 71 (EV71) strains isolated from specimens of patients with hand-foot-mouth disease (HFMD) in Ningxia province in 2008. Methods All the stool, throat swab and vesicle samples that collected from patients with HFMD were cultured. The positive isolates were identified by reverse transcriptase PCR (RT-PCR) with specific primers of EV71. Complete VP1 gene sequences (891 nucleotides) of 29 strains (part of 93 EV71 strains) were determined and compared with A, B and C genotype reference EV71 strains while EV71 China isolates by homogeneity and phylogenetic tree analyses. Results 215 strains of EV were isolated from 439 specimens. Results from RT-PCR indicated that 93 strains belong to EV71. Phylogenetic tree analysis revealed that the selected 29 stains were clustered with reference strains of C4 subgenotype. The nucleotide identity with C4 reference strains was 91.7%-99.4%. The amino acid homogeneity was 96.6%-100.0%. Conclusion The recently identified EV71 strains in Ningxia province belonged to subgenotype C4 which resembled to most of the isolates in China.

Objective To investigate the effect of Citronella oil on dementia of Alzheimer's type induced by D-galactose and aluminum trichloride (AlCl3) and explore its mechanism.Methods The 60 healthy KM mice with sex in half were enrolled in this study,10 of them were taken randomly as the normal group,the remainder 50 mice were in the model group.The rats received inntraperitoneal injection of D-galactose and oral aluminum trichloride for consecutive 10 weeks to construct the disease models.The model rats were randomly divided into five groups:model group,control group (0.5 g · kg-1 Piracetan) and three doses (0.4,0.6,0.8 mL · kg-1 Citronella oil) experimental groups.The ability of learning and memory was measured by the Morris water maze test.The levels of glutathione peroxidase (GSH-PX) and malondialdehyde (MDA) in serum homogenate and acetylcholine (ACh) in brain tissue homogenate were determined by spectrophotometer.Results At the fifth day in the Morris water maze training period,the escape latency(EL;s) in the normal group,model group,control group and three doses experimental groups were 21.42 ±-3.18,43.33 ± 2.14,26.84 ± 0.71,23.16 ± 3.70,18.48 ± 1.11,21.40 ±0.88.Compared with the normal group,the EL in model group was increased significantly (P ＜ 0.05).Compared with the model group,EL in control group and three doses experimental groups reduced significantly (P ＜ 0.05).The levels of GSH-PX and MDA in serum in the normal group,model group,control group and three doses experimental groups were (299.05 ± 90.62),(246.19 ± 37.90),(532.49 ± 121.52),(587.21 ± 177.37),(497.14 ±82.95),(493.33 ±73.64)U · mL-1;(9.38 ±3.01),(14.89 ±4.41),(6.98 ±2.43),(6.06 ±3.34),(5.26 ± 1.90),(5.01 ± 1.59) mmol · mL-1.Compared with the normal group,the GSH-PX and MDA in model group was decreased or increased significantly (all P ＜ 0.05).Compared with the model group,the GSH-PX and MDA in control group and three doses experimental groups reduced significantly (P ＜ 0.05,P ＜ 0.01).The level of ACh in brain tissue in the normal group,model group,control group and three doses experimental groups were (268.27 ±25.40),(224.95 ±21.52),(392.56 ±50.58),(240.17 ± 17.15),(255.50 ± 15.91),(259.15 ±20.74) μg · mL-1.Compared with the normal group,the ACh in model group was decreased significantly (P ＜ 0.01).Compared with the model group,the ACh in control group and middle and high doses experimental groups increased significantly (P ＜ 0.05,P ＜ 0.01).Conclusion Citronella oil,like Piracetam,enhanced the antioxidant activity of the Alzheimer's type induced by D-galactose and AlCl3,and increased the synthesis of ACh in brain.