Objective　To explore the effect of dexamethasone (Dex) given with the intention of prevention or treatment on endotoxin shock in rabbits and its relationship with tumor necrosis factor (TNF). Methods　Fifty-three health rabbits were divided into 4 groups, including normal control (n=13), endoxin shock group (n=16), preventive Dex group (n=12) and therapeutic Dex group (n=12). Except normal control was given with saline, the other 3 groups were administered with lipopolysaccharide (LPS) infusion, and the preventive Dex group was treated with Dex (5 mg/kg body weight) 30 min before LPS infusion and the therapeutic Dex group 20 min after LPS infusion. Mean arterial blood pressure (MABP), survival rate, TNF level in circulatory blood and other parameters were detected. Results　In preventive and therapeutic Dex groups, MABP was increased and survival rate was reduced compared with the animals from endoxin shock group (P<0.05, P<0.01), and TNF activity in the circulating blood was significantly suppressed (P<0.01). In addition, dexamethasone administration could alleviate the elevation of plasma glucagon, glucose, lactic acid, and β-glucironidase (P<0.05, P<0.01) in shocked animals. It was also found that administration of dexamethasone in vitro prevented the release of TNF by Kupffer cells. Conclusion　These results indicate that the preventive and therapeutic effect of dexamethasone on endotoxin shock, which may relate to its direct inhibition of the release of TNF induced by LPS.

OBJECTIVESTo seek a better profiling of proteins of hepatoma cells.

METHODSThe homogenate of hepatoma cells QGY-7703 was fractionated into four parts by differential centrifugation: the nuclei, the pellet by 20,000 x g, the pellet by 100,000 x g and the cytosolic supernatant. The four fractions were submitted to two-dimensional gel electrophoresis and their electrophoretic patterns were analyzed.

RESULTSIn comparison with the protein pattern of hepatoma cells not fractionated, the patterns of the four fractions display many more protein spots, and a large number of proteins present in the nuclei and cytosolic supernatant were not shown in the not-fractionated samples.

CONCLUSIONPreparation of subcellular fractions before electrophoretic procedures proves to be very useful; not only can it improve the results of two-dimensional gel electrophoresis, but also can lead to research into the subcellular level.

Carcinoma, Hepatocellular ; chemistry ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Liver Neoplasms ; chemistry ; pathology ; Neoplasm Proteins ; analysis ; Proteome ; Tumor Cells, Cultured

To study the difference of changes on apoptosis, necrosis and respiratory burst of the polymorphonuclear neutrophils (PMN) in endotoxemia rat model. LPS (O(55)B(5), 5 mg/kg) was injected into abdominal cavity of 20 random normal Wistar rat. 2, 4, 8 and 12 hours after injection, the changes of apoptosis, necrosis and respiratory burst of the rats between PMN from the peripheral blood and from the bronchoalveolar lavage fluid were observed using the flow cytometer. At the same time, 5 uninjected rats were taken as control. The results demonstrated that the quantity proportions of apoptosis of PMN between the peripheral blood PMN and the bronchoalveolar lavage fluid PMN in rat's endotoxemia were similar. However, comparison with the uninjected LPS rat, the necrosis of peripheral blood PMN obviously increased and the respiratory burst capacity was clearly inhibited. Contrarily, the necrosis of bronchoalveolar lavage fluid PMN obviously decreased and the respiratory burst obviously increased in the injecting LPS rat. It was concluded that the necrosis and apoptosis displayed differently between the pulmonary and peripheral blood PMNs in endotoxemia. Under state of inflammation, the surviving PMN in tissue increased and kept the activated state due to tissue injury.
Animals ; Apoptosis ; Bronchoalveolar Lavage Fluid ; cytology ; Endotoxemia ; blood ; Necrosis ; Neutrophils ; physiology ; Pulmonary Alveoli ; pathology ; Rats ; Rats, Wistar ; Respiratory Burst

BACKGROUNDPolymorphonuclear neutrophil (PMN), one of the most important inflammatory cells, functions throughout the initiation, progression and resolution of inflammation. This study aimed at investigating the relationship between PMN apoptosis and the lung injury after chest impact trauma.

METHODSPMNs were purified from rabbits subjected to the chest impact trauma and their apoptosis, necrosis, survival and respiratory burst were detected by flow cytometry. Meanwhile, lactate dehydrogenase and (LDH) [Ca2+]i were measured.

RESULTSThe delayed apoptosis of PMNs in bronchoalveolar lavage fluid was observed from 2 hours to 12 hours after trauma, and viable cells increased. Respiratory burst of PMNs in bronchoalveolar lavage fluid was increased significantly from 2 hours with the peak at 8 hours. Meanwhile, lactate dehydrogenase in bronchoalveolar lavage fluid was higher than that in control (P < 0.05) from 4 hours to 24 hours, and intracellular free Ca2+ in PMN was increased temporarily.

CONCLUSIONSRetention of PMN in tissues and the abnormality in apoptotic pathway inevitably generate persistent activation of PMN and excessive release of toxic substances, resulting in tissue injury. The temporary increase of intracellular free Ca2+ may be responsible for the delayed apoptosis of PMN.

Animals ; Apoptosis ; physiology ; Lung Injury ; Neutrophils ; physiology ; Rabbits ; Respiratory Burst ; physiology ; Thoracic Injuries ; complications

BACKGROUND: Amniotic membrane has a unique structure that can block the penetration of certain substances, to ensure normal nutrition supply for the surrounded tissues, and is also characterized by anti-adhesion, good tissue compatibility, mild inflammatory reaction, few fibers and biodegradability. OBJECTIVE: To compare the effects of fresh amniotic membrane and acellular amniotic membrane to prevent adhesion and promote tendon healing during the repair of tendon sheath defects. METHODS: Sixty healthy male leghorn chickens were selected, and the model of tendon injury and tendon sheath defects was established at the third toes. The animal models were then randomly divided into three groups and underwent repair with fresh amniotic membrane (group A), acellular amniotic membrane (group B), and no treatment in control group (group C), respectively. Histological observation and biomechanical analysis of the third toes were performed after repair. RESULTS AND CONCLUSION: (1) Histological observation. Congestive edema and inflammatory response were found in all animals at 2 weeks after repair, but mildest in the group A and severest in the group C. These inflammatory responses gradually alleviated over time in the three groups. At 12 weeks after repair, the new tendon sheath formed in all the animals, which was more mature than that at 4 weeks after repair. The synovial cells on the surface of the tendon sheath were arrayed tidily with dense structure in the groups A and B, but in the group C, the synovial cells were distributed disorderly with loose structure and prominent fibrous tissues. (2) Biomechanical analysis. Tendon sliding distance in the groups A and B was significantly larger than that in the group C at 4, 8, 12 weeks after repair (P < 0.05), but there were no significant difference in the distance between the groups A and B (P > 0.05). At 4 and 8 weeks after repair, the maximum tensile strength was largest in the group A, sequentially followed by group B and group C (P < 0.05), but there were no significant difference among the three groups at 12 weeks after repair (P > 0.05). To conclude, both fresh amniotic membrane and acellular amniotic membrane can promote tendon healing and prevent the adhesion of tendon through tendon sheath reconstruction, but the fresh amniotic membrane is preferred to promote early tendon healing compared with acellular amniotic membrane.

The protective effects of cyclosporin A (CsA), an inhibitor of mitochondrial permeability transition pore (MPTP), on vascular permeability in sepsis rats were investigated. Cecal ligation and puncture (CLP)-induced sepsis rats were used for in vivo studies, and the effects of CsA (1 and 5 mg·kg^-1) on vascular permeability of lung, kidney, and intestine, mitochondrial respiratory control ratio, and the survival of the sepsis rats were observed. Lipopolysaccharide (LPS) was used for stimulating vascular endothelial cells (VECs) in vitro, and the effects of CsA on leakage of microvascular, immunofluorescence of zonula occludes-1 (ZO-1), and transendothelial electrical resistance (TER) were observed. All the animal welfare and experimental procedures are in accordance with the regulations of the Animal Ethics Committee of the Army Medical University. Compared with sham-operated group, the vascular permeability of lung, kidney, and intestine in sepsis rats increased significantly (P<0.05). Compared with conventional treatment group, CsA could significantly decrease the vascular permeability of lung, kidney, and intestine (P<0.05 or P<0.01), and prolong the survival period. The results of microcirculation also showed that CsA could significantly reduce the permeability of mesenteric venules in sepsis rats. At the cellular level, LPS stimulation significantly increased the permeability of vascular endothelial cells, including the decrease of transmembrane resistance and protein expression of ZO-1 (P<0.05). CsA can significantly reduce the increase of permeability of vascular endothelial cells induced by LPS stimulation (P<0.01). The function of mitochondria in the kidneys and intestines of sepsis rats was obviously impaired, and the respiratory control ratio of mitochondria was decreased. LPS significantly increased MPTP opening of VECs, while CsA significantly inhibited MPTP opening and improved mitochondrial function. CsA may protect mitochondrial function by inhibiting the opening of MPTP and play a protective role in the vascular permeability of sepsis rats. This study will provide an insight for the treatment of sepsis vascular leakage.

In order to study the pesticide residues of the medicinal Crataegi Fructus,this study aims to establish an analysis method for pesticide residues( mainly containing insecticides and fungicides) suitable for the actual situation of medicinal Crataegi Fructus based on the survey of the pesticides of the Crataegi Fructus base,combined with the blind screening results of the LC-ESI-MS/MS pesticide screening platform established by the research team in the early stage. Then,the pesticide residues in medicinal Crataegi Fructus from Shandong,Hebei,Henan,Shanxi,and Liaoning( main cultivation areas) were analyzed. The samples were pretreated by the modified Qu ECh ERS method,i.e.,extracted with acetonitrile-water( 9 ∶1),purified by PSA,C_(18),GCB,silica gel. The detection of pesticides was performed by LC-MS/MS. The ion source was ESI with positive scanning mode,and the linearity of 11 kinds of pesticides in the range of 5-300 μg·kg~(-1) was acceptable( R~2>0. 996 9). All the recoveries of pesticides were within 70. 02%~(-1)12. 0% in the low,medium and high levels,with RSD≤17%. The results showed that the detection rate of carbendazim,chlorpyrifos and difenoconazole is 79%,82%,56%,respectively. Besides,the prohibition pesticide carbofuran were detected in some of the batches,indicating the security risk. This study provides methodological references and basic data for risk assessment of Crataegi Fructus and government regulation.
Chromatography, Liquid ; Crataegus/chemistry* ; Drug Contamination ; Drugs, Chinese Herbal/analysis* ; Pesticide Residues/analysis* ; Surveys and Questionnaires ; Tandem Mass Spectrometry