Objective To investigate the effects of inducible hypothermia on expressions of peroxisome proliferator activated receptor gamma coactivator-1 (PGC-1) and small heterodimer partner (SHP) in rat model of hemorrhagic shock.Methods SD rats were randomly (random number) divided into three groups:control (C),normothermia (N) and hypothermia (H).Exsanguination was carried out in rats by continuously drawing out venous blood (25 mL/kg) over 15 minutes to establish hemorrhagic shock model.Then,rectal temperatures of rats were maintained by body surface cooling to 32℃ in H group and by body surface warming to 38℃ in N group,respectively.After a shock period of 60 minutes,rats received the infusion of whole blood of their own and lactated Ringer's solution (1 ∶ 2) treatment for 60 min.Rats were warmed to 38℃ by body surface warming and monitored for 3 h after resuscitation.Hematocrit (Hct),base excess (BE),lactate (Lac),and glucose (Glu) were recorded before modeling and after different lengths of hemorrhagic shock period (HSP).The expressions of PGC-1 mRNA and SHP mRNA and the levels of their protein in liver were tested by real-time reverse transcription polymerase chain reaction (qRT-PCR) and western blotting,respectively.Results The H group had lower lactate levels and higher BElevels than the N group [(6.3±2.1) vs.(10.4±1.5) and (-4.7±2.5) vs.(-9.0±3.2)] (P＜ 0.05).At 72 hours after modeling,there were four survivors in the N group and seven survivors in the H group (P ＜ 0.05,Log Rank).The expressions of PGC-1 mRNA and SHP mRNA increased in N group.Hypothermia resuscitation down-regulated PGC-1 mRNA expression,meanwhile,increased expression of SHP mRNA.Both Hypothermia and Normothermia resuscitation increased SHP protein levels,but decreased PGC-1 protein levels.Conclusions Inducible hypothermia ameliorated acidosis and energy metabolism imbalance through adaptive regulation in PGC-1 and SHP.

Objective To observe the effect of Yangxue Qingnao Granule on the ultrastructures and expression of p38 mitogen activated protein kinases (p38MAPK) in CA1 area of hippocampus in vascular dementia rats. Methods 180 Sprague-Dawley rats were randomly di-vided into sham group, vascular dementia model group (model group) and Yangxue Qingnao Granule treatment group (treatment group). The vascular dementia was modeled with modified Pulsineli's four-vessel occlusion. The ultrastructure of CA1 area was observed with trans-mission electron microscope, while the expression of p38MAPK in CA1 area was detected with immunohistochemstry and Western blotting 1, 2, 4 and 8 weeks after modeling. Results In the model group, pyknosis, nuclear dissolution, heterochromatin margination and mitochon-dria swelling were found in most of the neurons in CA1. In the treatment group, the distribution of chromatin was well-proportioned, and mi-tochondrion and other organelle were normal. In the model group, the expression of p38MAPK increased at each time point compared with the sham group (P<0.01), and peaked 4 weeks after modeling, and decreased in the treatment group compared with the model group (P<0.01). Conclusion Yangxue Qingnao Granule can improve the ultrastructure of neuronal in CA1 area of hippocampus of vascular dementia rats, which may relate with the inhibition of the expression of p38MAPK.

This study was aimed to observe the effect of Yang-Xue Qing-Nao(YXQN) granules on the expression of brain-derived neurotrophic factor (BDNF) and basic fibroblast growth factor (bFGF) in CA1area of hippocampus among vascular dementia (VD) rats.A total of 72 SD rats were randomly divided into the sham operation group,VD model group (model group) and YXQN treatment group (treatment group).The VD rat model was prepared by modified Pulsineli's four-vessel occlusion.The learning and memory abilities of rats were detected by the Morris water maze.The protein expressions of BDNF and bFGF were detected by the immunohistochemical analysis.The results showed that compared with the sham operation group,there was obvious learning and memory disorders in the model group with increased protein expressions of BDNF and bFGF in CA1 area of hippocampus at difference time points (P<0.01).Compared with the model group,the learning and memory abilities of rats in the treatment group were significantly improved; and the protein expressions of BDNF and bFGF in CA1 area of hippocampus were significantly increased at difference time points (P<0.01).The expression was the highest on the 4th week.It was concluded that YXQN granules can improve the learning and memory abilities of VD rats.Its mechanism may be related to the upregulation of protein expression of BDNF and bFGF.

OBJECTIVETo study the expression of heme oxygenase-1 (HO-1) gene and protein and the effect of budesonide (BUD) on the HO-1 expression in lung tissues in rats with asthma.

METHODSFifty male Sprague-Dawley rats were randomly divided into 5 groups: normal control, asthma model, dexamethasone (DXM)-, hemin (HO-1 challenger)-or BUD-treated asthma. The asthma model was prepared by ovalbumin sensitization and challenge. The rats were sacrificed 24 hrs after the last challenge. The blood COHb content,and the total cell count and the percentage of differential cells in bronchoalveolar lavage fluid (BALF) were measured. The expression of HO-1 protein and mRNA in lung tissues was detected with immunohistochemistry and RT-PCR, respectively. The airway inflammation situations were evaluated by histopathology.

RESULTSThe airway inflammatory cell infiltration in the DXM-, hemin- and BUD-treated asthma groups was remarkably alleviated compared with that in the asthma model group. Compared with the normal control group, the expression of HO-1 mRNA and protein in lung tissues and the blood COHb content in the asthma model and the DXM-, hemin- and BUD-treated asthma groups were significantly up-regulated. The DXM-, hemin- and BUD-treated asthma groups showed significantly increased expression of HO-1 protein and mRNA in lung tissues compared with the asthma model group. The blood COHb content in the DXM-and the hemin-treated asthma groups was significantly higher than that in the asthma model group.

CONCLUSIONSThe expression of HO-1 protein and mRNA in lung tissues and blood HO-1 activity increased in rats with asthma,suggesting that HO-1 may be involved in the pathogenesis of asthma. HO-1 may have a protective effect against the airway inflammation in asthmatic rats. BUD and DXM can up-regulate the expression of HO-1 protein and mRNA, thus providing protective effects against the airway inflammation in asthmatic rats.

Animals ; Asthma ; drug therapy ; enzymology ; Budesonide ; pharmacology ; therapeutic use ; Carboxyhemoglobin ; analysis ; Dexamethasone ; pharmacology ; Heme Oxygenase-1 ; analysis ; genetics ; Hemin ; pharmacology ; Lung ; enzymology ; Male ; RNA, Messenger ; analysis ; Random Allocation ; Rats ; Rats, Sprague-Dawley

BACKGROUND: Clinical evidence have proved that heterotopic ossification is easily present in bony callus of patients with fracture combined with head injury, and the healing of bone fracture is accelerated obviously. OBJECTIVE: To study the expression and distribution of vascular endothelial growth factor (VEGF) in bony callus among the patients with fracture combined with head injury and simple fracture patients, and to investigate its clinical significance and action mechanism.DESIGN, TIME AND SETTING: Grouping controlled observation was performed in the Xijing Hospital, the Fourth Military Medical University of Chinese PLA between February 2006 and July 2007.PARTICIPANTS: The patients comprised two groups those who had fracture combined with head injury and those who had simple fracture. Each group had 50 patients. The fracture combined head injury group included 41 males and 9 females, aged 19-55 years; simple fracture group included 36 males and 14 females, aged 17-52 years. METHODS: The bony callus specimens were harvested at 7-10, 11-15, 16-20, 21-27 and 28-35 days post-injury. The content of VEGF in bony callus was measured at different phase by using immunohistochemistry methods, and the speed of fracture healing was also observed.MAIN OUTCOME MEASURES: ①X-ray results; ②immunohistochemical image analysis. RESULTS: In different phase of bone healing, the VEGF of the two groups were homogenous, the early expression of VEGF in osteoprogenitor cell, osteoblast and cartilage cell in the fracture combined with head injury group was significantly higher than those in simple fracture group. The osteoprogenitor cell proliferated, and the differentiations of osteoblast and cartilage cell were enhanced obviously. The VEGF level reached the peak at 7-10 days, and kept at the high level for 30 days and then gradually decreased. The peak of VEGF level in simple fracture group came at 11-15 days, kept in the high level for 20 days and then gradually decreased. The peak value in simple fracture group was significantly lower than the fracture combined with head injury group. There was statistical significance in the expression of VEGF between two groups by the paired t-test (P < 0.05). A large mount of osteotylus could be detected at 4 weeks after injury by X-ray in patients with fracture combined, with head injury, but at 7-9 weeks in patients of those who had simple fracture.CONCLUSION: The expression of VEGF in the bony callus in the group of fracture combined with head injury is significantly greater than that in simple fracture patients, and the expression lasts for a longer time. The proliferation and differentiation of osteogenitor call, osteoblast and cartilage cell are also significantly faster in the group of fracture combined with head injury, which might be one of the bone healing mechanisms.

Humans ; In Situ Hybridization, Fluorescence ; Membrane Proteins ; analysis ; NF-kappa B ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Tissue Array Analysis ; methods ; Tumor Cells, Cultured

This study was aimed to observe the effect ofYang-Xue Qing-Nao(YXQN) granules on expressions of glycogen synthase kinase-3β (GSK-3β) andβ-catenin in CA1 area of hippocampus among vascular dementia (VD) rats. A total of 72 SD rats were randomly divided into the sham operation group, VD group (model group) and the YXQN granules treatment group (treatment group). The VD rat model was prepared by modified Pulsineli’s four-vessel occlusion. The expressions of GSK-3β andβ-catenin in CA1 area of hippocampus were detected by immunohistochemistry and western blotting at the 1st week, 2nd week, 4th weeks and 8th week after VD model operation. The results showed that expressions of GSK-3βwere increased in the model group at different time points, which were many quantities of expression at the 1st week, and a large number of expressions at the 2nd week. It reached peak at the 4th week; and began to decline but still higher at the 8th week. Compared with the sham operation group, the expression of GSK-3β was significantly increased in the model group at different time points (P < 0.01). The expression ofβ-catenin was increased in the model group at different time points. However, there was no statistical significance compared with the sham operation group (P > 0.05). Compared with the model group, expressions of GSK-3β andβ-catenin were significantly increased in the treatment group at different time points (P < 0.01). It was concluded that YXQN granules upregulated the expression of GSK-3β andβ-catenin, which may be helpful to VD treatment.

OBJECTIVETo establish a simple and accurate method for rapid detection of lamivudine related mutations in hepatitis B virus (HBV) polymerase gene.

METHODSHBV polymerase gene fragments of covering B and C active region were amplified by nested polymerase chain reaction (nPCR) or nested mismatched PCR. The PCR products were digested with Nde I or Nia III and subjected to electrophoresis on agarose gel, respectively. The patterns of restriction fragment length polymorphism (RFLP) were distinguished. Using this method, thirty patients with chronic hepatitis B and treated with lamivudine for at least one year were analysed for the lamivudine related mutations in polymerase gene. Sixteen cases without lamivudine therapy were used as controls. Some of the patients were also analysed by clone sequencing.

RESULTSThe nested mismatched PCR-RFLP method was simple, accurate and rapid. The whole experiments could be finished in eleven hours. The least titers of HBV DNA which could be detected was 10.3 copies/ml. The wild or mutant strains judged by RFLP were identified by clone sequencing. Mutation in the tyrosine methionine aspartic aspartic acid (YMDD) motif of HBV polymerase gene was found in eight patients and mutations of YMDD motif associated with L526M were found in another three patients. However, there were no such mutations in the control cases.

CONCLUSIONThe nested PCR-RFLP is considered as a simple and accurate method for rapid detection of lamivudine related mutations in HBV polymerase gene. It is suitable for larger number of sample detection.

Antiviral Agents ; pharmacology ; DNA-Directed DNA Polymerase ; genetics ; Drug Resistance, Viral ; genetics ; Hepatitis B virus ; drug effects ; enzymology ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; pharmacology ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length

The purpose of the present study is to use beta-cyclodextrin polymers (beta-CDP) with different cross-linked degree (CLD) to form inclusion complexes with ibuprofen and examine the effects of structural and compositional factors of beta-CDP on its drug loading and release behaviors. A series of beta-CDP with different CLD were synthesized and characterized by Fourier Transform Infrared Spectroscopy (FT-IR) and 13C NMR spectrum. The beta-CDP was systemically characterized for the relation between the CLD of beta-CDP and the drug loading and release as well. The results of FT-IR and 13C NMR showed that similar peak-shaped vibration of beta-CDP and beta-CD implies that the polymer keeps the original characteristic structure of beta-CD. The CLD of the beta-CDP played a critical role in the drug loading and release, increasing the CLD resulted in reduction of drug loading, but increase in drug release.
Carbon Isotopes ; Cross-Linking Reagents ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Ibuprofen ; administration & dosage ; chemistry ; Magnetic Resonance Spectroscopy ; Pharmaceutical Preparations ; administration & dosage ; chemistry ; Polymers ; chemistry ; Solubility ; Spectroscopy, Fourier Transform Infrared ; beta-Cyclodextrins ; chemistry

Objective To study the COLIXA3 gene polymorphism of patients with fluorosis and to explore the pathogenesis of COLIXA3 gene in endemic fluorosis.Methods Fifty one cases of patients with drinking-water borne fluorosis were selected as the case group in Xinzhou city,Shanxi province and 28 cases of healthy people were as the control group.Dental fluorosis was detected by Dean method and skeletal fluorosis was examined by X-ray.COLIXA3 of exon 5 gene product of 103 points was amplified by PCR and the gene locus genotype was sequenced.Results Ten cases of mild dental fluorosis,14 cases of moderate dental fluorosis,15 cases of severe dental fluorosis were detected among the 51 patients.The control group was free of dental fluorosis.All the 51 cases of patients with fluorosis had varying degrees of skeletal fluorosis,mainly osteosclerosis lesions,accounting for 86.27％(44/51 ),and mild skeletal fluorosis patients were all osteosclerosis lesions,and osteosclerosis lesions and multiple skeletal lesions were found among moderate and severe skeletal fluorosis patients in the case group,while control group had no skeletal fluorosis.The differences between genotypes of frequency distribution of AA,Aa,aa of COLIXA3 of case and control groups were not statistically significant [96.08％(49/51 ),3.92％(2/51 ),0.00％(0/51) and 96.43％(27/28),3.57％(1/28),0.00％(0/28),x2 =0.94,P ＞ 0.05].ConclusionsCOLIXA3 gene polymorphism is not significantly correlated to fluorosis.