OBJECTIVE: To observe the changes of malondialdehyde (MDA), superoxide dismutase (SOD), tumor necrosis factor-alpha (TNF-alpha), and interleukin-1beta (IL-1beta) in rat brain tissue and to explore the mechanism of secondary cerebral injury after brain concussion. METHODS: The brain concussion model was established with the pathological changes of rat brain tissue by Weil stain. The expressions of MDA and SOD in brain tissue were examined by photochemical method. The expressions of TNF-alpha and IL-1beta in cerebral cortex and hippocampus were examined by immunochemistry. RESULTS: Nerve myelin sheath showed disorder, disruption, gryposis and swelling by Weil stain. Above changes were more severe at 12h. The quantity of MDA in rat brain tissue after concussion was significantly higher than that in the control group. The activity of SOD was significantly lower than that in the control group. The expressions of TNF-alpha and IL-1beta increased more significantly in cerebral cortex and hippocampus in rat brain tissue after concussion than that in the control group. CONCLUSION Oxidative stress and inflammatory injury in the rat brain tissue, which may play an important role in secondary cerebral injury after concussion.
Animals ; Brain/metabolism* ; Brain Concussion/metabolism* ; Brain Injuries ; Hippocampus ; Interleukin-1beta/metabolism* ; Malondialdehyde/metabolism* ; Oxidative Stress ; Rats ; Superoxide Dismutase/metabolism* ; Tumor Necrosis Factor-alpha/metabolism*

The medium of photo-synthetic bacteria synthesizing carotenoid was optimized with response surface methodology. The regression equation expressing the relationship between carotenoid yield and main components of culture medium was established by stepwise analysis. The R-squared in the model of regression equation was 0. 958, which meant the established equation could predict the carotenoid yield well at the range of factors in this design. It was indicated that the optimum medium components were: 0. 81% citric acid, 0. 35% NH4Cl and 0. 18% corn syrup. On those conditions, it was predicted that the highest production of carotenoid was 13. 34 mg/L, which was 2. 04 times higher than initial carotenoid yield before optimization.

OBJECTIVE: To investigate the role of endoplasmic reticulum stress (ERS) in lipopolysaccharide (LPS)-induced hepatocyte apoptosis. METHODS: Cells of the rat hepatocyte line BRL were cultured. The hepatocytes were treated with LPS, ERS inducer thapsigargin (TG), and ERS inhibitor 4-phenylbutyric acid (4-PBA), respectively or in their different combination. The cell viability was measured by MTT assay. The cyto-nuclear morphological changes of apoptosis cells were detected by the fluorescent dye Hoechst 33258. The apoptosis rate was assessed by flow cytometry with Annexin V-FITC/PI double-staining. Expressions of GRP78 as ERS marker protein, CHOP, caspase-12 and cleaved-caspase-3 as ERS related protein were detected by Western blotting. RESULTS: LPS could cause a decrease in cell viability and an increase in apoptosis rate in a dose- and time-dependent manner. The expression of GRP78, CHOP, caspase-12 and cleaved-caspase-3 proteins were significantly increased with LPS treatment. TG led to a marked decrease in cell viability and an increase in apoptosis rate, which aggravated the hepatocyte injury induced by LPS; whereas 4-PBA alleviated LPS-induced apoptosis. CONCLUSION ERS mediates LPS-induced hepatocyte injuries, indicating that ERS may play a vital role in the pathogenesis of LPS-induced hepatocyte injuries.
Animals ; Apoptosis ; Caspase 3 ; Cell Survival ; Endoplasmic Reticulum Chaperone BiP ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; Hepatocytes ; Lipopolysaccharides ; Phenylbutyrates ; Rats

In practice of forensic medicine, potential disease can be associated with fatal asphyxia in restraint position. Research has demonstrated that nitric oxide (NO) and nitric oxide synthase (NOS) are plentifully distributed in skeletal muscle, contributing to the regulation of contractile and relaxation. In the current study, respiratory functions, indices of diaphragmatic biomechanical functions ex vivo, as well as NO levels in serum, the expressions of diaphragmatic inducible NOS (iNOS) mRNA, and the effects of L-NNA on contractility of the diaphragm were observed in sepsis induced by cecal ligation and puncture (CLP) under the condition of restraint position. The results showed that in the CLP12-18h rats, respiratory dysfunctions; indices of diaphragmatic biomechanical functions (Pt, +dT/dt(max), -dT/dt(max), CT, Po, force over the full range of the force-frequency relationship and fatigue resistance) declined progressively; the NO level in serum, and iNOS mRNA expression in the diaphragm increased progressively; force increased significantly at all stimulation frequencies after L-NNA pre-incubation. Restraint position 1 h in CLP12 h rats resulted in severe respiratory dysfunctions after relative stable respiratory functions, almost all the indices of diaphragmatic biomechanical functions declined further, whereas little change took place in NO level in serum and diaphragmatic iNOS mRNA expression; and the effects of L-NNA were lack of statistical significance compared with those of CLP12 h, but differed from CLP18 h group. These results suggest that restraint position and sepsis act together in a synergistic manner to aggravate the great reduction of diaphragmatic contractility via, at least in part, the negative modulation of NO, which may contribute to the pathogenesis of positional asphyxia.
Animals ; Asphyxia ; Diaphragm/physiology* ; Muscle Contraction ; Muscle, Skeletal ; Nitric Oxide/metabolism* ; Nitric Oxide Synthase ; Nitric Oxide Synthase Type II ; Rats ; Respiration Disorders ; Restraint, Physical ; Sepsis

OBJECTIVETo evaluate clinical results of early total care (ETC) treatment for elderly patients with intertrochanteric femur fractures.

METHODSClinical data of 106 elderly patients with intertrochanteric fracture treated from January 2012 and February 2015 were retrospectively studied. According to whether receiving the early total care mode, the patients were divided into 2 groups, 34 cases were diagnosed and treated with early total care pattern (ETC group), including 14 males and 20 females with an average age of (74.88 ± 4.38) years old ranging from 70 to 86. According to Evans types, 4 cases were type I, 5 cases were type II, 13 cases were type III, 11 cases were type IV, 1 case was type V. Seventy-two patients were treated with conventional trauma method (conventional group), including 35 males and 37 females with an average age of (74.46 ± 3.63) years old ranging from 70 to 85. According to Evans type, 8 cases were type I ,13 cases were type II, 25 cases were type III, 25 cases were type IV, and 1 case was type V. All fractures were treated with proximal femoral nails anti-rotation (PFNA). Operative time, hospital stays, leaving bed time, complications, cases of death at 1 year after operation, postoperative Harris score at 12 months were observed and compared.

RESULTSAll patients were followed up, the time of ETC group ranged from 9 to 18 months with an average of 13.29 ± 1.51, and the time in conventional group ranged from 12 to 16 months with an average 12.93 ± 1.15, while there was no significant difference between two groups in time of following-up (t = 1.368, P = 0.174). There was no significant meaning in cases of death between ETC group (2 cases) and conventional group (8 cases). Three cases occurred complications in ETC group, and 20 cases in conventional group,there was obvious meaning between two groups (χ² = 0.739, P = 0.318). Operative time,hospital stays,leaving bed time in ETC group respectively was (2.03 ± 0.67) d, (15.41 ± 2.87) d and (3.62 ± 0.74) d; while in conventional group respectively was (4.17 ± 1.59) d, (20.11 ± 4.24) d and (5.35 ± 1.22) d; there were significant differences between two groups in operative time, hospital stays, leaving bed time. Postoperative Harris scores at 12 months in ETC group was (82.32 ± 4.56), and (79.24 ± 5.52) in conventional group, there was obvious meaning between two groups (t = 2.833, P = 0.006).

CONCLUSIONETC pattern is a novel method for diagnosis and treatment of intertrochanteric femur fractures in elderly, it could shorten operative time, hospital stays, leaving bed time, decrease complications and promote recovery of function.

Aged ; Aged, 80 and over ; Female ; Femur ; injuries ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; Hip Fractures ; surgery ; Hip Joint ; surgery ; Hospitalization ; Humans ; Male ; Operative Time ; Postoperative Care ; Retrospective Studies

Objective To explore the effect of uhrasound-phoresis of Chinese herb in the treatment of frac- ture healing on rats.Methods The animal model of femur fracture was established in 36 rats,who were divided in- to 4 groups after operation.The rats in the ultrasound group were given ultrasound treatment daily.The herbal group was given Chinese herb applied on the fracture site.The experimental group was given uhrasound-phoresis of Chinese herb on the site of fracture.The control group was housed without any treatment.All rats were sacrificed at 30 days and the bony callus were harvested and observed with histological anti immunohistoehemical examination.Results The histological examination showed that the appearance of cartilaginous and bony callus in the experimental group were earlier than those in other groups,Immunohistochemical examination showed that the expression of collagenⅠin the experimental group was significantly higher than that in ultrasound group and herbal group(P

OBJECTIVE: To observe the time-course expression of calcium-calmodulin dependent protein kinase II delta (CaMK II delta) in cerebral cortex after traumatic brain injury (TBI). METHODS: The TBI rat model was established. The expression of CaMK II delta in cerebral cortex around injured area was tested by Western blotting and immunohistochemical staining. RESULTS: Western blotting revealed expression of CaMK II delta in normal rat brain cortex. It gradually increased after TBI, peaked after 3 days, and then returned to normal level. The result of immunohistochemical staining was consistent with that of Western blotting. CONCLUSION The expression of CaMK II delta around injured area after TBI increased initially and then decreased. It could be used as a new indicator for wound age determination following TBI.
Animals ; Blotting, Western ; Brain Injuries/metabolism* ; Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism* ; Cerebral Cortex/metabolism* ; Forensic Medicine ; Immunohistochemistry ; Rats ; Time Factors

OBJECTIVE: To explore the effect of nitric oxide (NO) on the gene expression of hepatic TNF-α and IL-1β by crush injury of rat's soft tissues. METHODS: Rats were randomly divided into sham group, crush group, crush+aminoguanidine (AG) group, and crush+L-arginine (L-Arg) group. Activities of ALT and AST as well as NO level in serum were measured. Gene expressions of TNF-α and IL-1β were detected with RT-PCR. RESULTS: Obvious increase in TNF-α and IL-1β mRNA expression was detected in the crush group compared with the sham group (P<0.05). After pretreated L-Arg, expressions of TNF-α and IL-1β mRNA were markedly increased (P<0.05). After pretreated AG, those indices obviously decreased (P<0.05). Activities of ALT and AST enhanced and NO level increased in the crush group compared with the sham group (P<0.05). Pretreatment with L-Arg or AG led to substantial increased or reduced activities of ALT and AST as well as NO levels, respectively. CONCLUSION Endogenous NO mediated TNF-α, IL-1β mRNA up expression in liver induced by increased production of NO after crush injury of rat's soft tissues.
Animals ; Gene Expression ; Interleukin-1beta/metabolism* ; Liver ; Nitric Oxide/physiology* ; RNA, Messenger ; Rats ; Tumor Necrosis Factor-alpha/metabolism* ; Wounds and Injuries

OBJECTIVE: To investigate effects of antioxidant stress protein heme oxygenase-1 (HO-1) on lipopolysaccharide (LPS)-induced endoplasmic reticulum stress (ERS) of rat hepatocytes. METHODS: The BRL cells (rat hepatocyte cell line) were cultured. The hepatocytes were treated with LPS, LPS+HO-1 siRNA, HO-1 siRNA and PBS solution, respectively. The cell viability was measured by trypan blue exclusion test. The apoptosis cells were detected by the fluorescent dye Hoechst 33258. Expressions of GRP78, CHOP, caspase-12 and HO-1 were detected by Western blotting. RESULTS: LPS caused an increase of HO-1 protein expression of rat hepatocytes in a dose-dependent and time-dependent manner, a up-regulation of GRP78, CHOP and caspase-12, a decrease in cell viability, and an increase in apoptosis rate of hepatocytes. Pretreatment of HO-1 siRNA inhibited the up-regulation of LPS-induced HO-1, however, aggravated ERS and cellular injury. CONCLUSION HO-1 inhibites ERS-mediated cellular injury of rat hepatocytes induced by LPS.
Animals ; Apoptosis/physiology* ; Endoplasmic Reticulum/metabolism* ; Endoplasmic Reticulum Stress/physiology* ; Heme Oxygenase-1/pharmacology* ; Hepatocytes/metabolism* ; Lipopolysaccharides/pharmacology* ; Rats

OBJECTIVE: To explore the role of hydrogen sulfide (H2S) in acute liver injury induced by crushing hind limbs of rats. METHODS: The rats were randomly divided into the following groups: control, crushing, H2S donor sodium hydrosulfide (NaHS) + crushing, H2S inhibitor propargylglycine (PAG) + crushing group. The acute liver injury model was established by 'crushing the hind limbs of rats with standard weight. Rats were sacrificed at 30 min and 120 min after the crush. The activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured by colorimetric method, and the content of H2S in plasma and the contents of malondialdehyde (MDA), protein carbonyl, glutathione (GSH) in the liver and the activity of H2S generating enzyme (cystathionine y-lyase, CSE) were determined by chemical method. The expression of CSE mRNA in liver was detected by RT-PCR. RESULTS: For crush injury group, the levels of AST and ALT in serum, MDA and protein carbonyl in liver increased. The levels of GSH, CSE, CSE mRNA in liver and H2S in serum decreased. The administration of NaHS before limbs crush could attenuate the changes of liver injury, but the pre-treatment with PAG could exacerbate the changes. CONCLUSION The decrease of H2S production could involve in mediating the acute liver injury induced by traumatic stress in rats.
Alanine Transaminase/blood* ; Alkynes/pharmacology* ; Animals ; Aspartate Aminotransferases/blood* ; Cystathionine gamma-Lyase/metabolism* ; Glutathione/metabolism* ; Glycine/pharmacology* ; Hydrogen Sulfide/pharmacology* ; Liver/injuries* ; Malondialdehyde/metabolism* ; Protein Carbonylation ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sulfides/pharmacology*