Objective To evaluate the mitochondrial function of chondrocytes in Kashin-Beck disease (KBD) patients and its correlation with age.Methods Mitochondrial function was evaluated by analyzing the activities of respiratory chain enzyme complexes and citrate synthase (CS),intracellular adenosine triphosphate (ATP)contents, changes in mitochondrial membrane potential (ΔΨm),as well as the intracellular reactive oxygen species (ROS) contents.Correlation of mitodhondrial function and age was analyzed with linear regression.Results Activities of complexes Ⅱ (P = 0.001 ),Ⅲ (P = 0.005 )and Ⅳ (P = 0.005 )were reduced in KBD articular chondrocytes compared with cells from normal controls.However,the mitochondrial mass was increased in KBD samples. Cultured KBD chondrocytes had reduced cellular ATP level (P = 0.001 )and mitochondrial membrane potential (P =0.001),but increased citrate synthase activity (P =0.04)and intracellular ROS level (P <0.001).We found no correlation between mitochondrial function and donor age either in normal or in KBD chondrocytes. Conclusion Mitochondrial function is altered in articular chondrocytes of KBD,and no correlation was found between mitochondrial function and donor age.

Asian Continental Ancestry Group ; genetics ; Carcinoma, Hepatocellular ; genetics ; China ; Gene Frequency ; Genetic Predisposition to Disease ; Humans ; Liver Neoplasms ; genetics ; Mongolia ; Polymorphism, Single Nucleotide

Objective To examine the changes of serum total and high density lipoprotein cholesterol during whole blood incubation and to investigate the possible mechanisms responsible for the instability.Methods A method for the measurement of serum total cholesterol (TC), total free cholesterol (TFC), high density lipoprotein cholesterol (HDLC) and high density lipoprotein free cholesterol (HDLFC) by high performance liquid chromatography (HPLC) was developed. Whole blood specimens were incubated at 25℃ and 4℃ for various period of time and serum TC, TFC, HDLC and HDLFC were measured.Results The new HPLC method showed a mean within-run CV of 0.22%~0.51%. The averaged changes during the incubations ranged 0.6%~2.0% for TC and 2.0%~4.1% for HDLC.Conclusion An HPLC method has been established that is highly precise and can be used for detecting subtle cholesterol changes in biological samples. Different cholesterol exchanges or transfers between blood cells and lipoproteins exist during various incubations. Prolonged whole blood storage that causes serum TC and HDLC changes should be avoided in clinical lipid measurements.

Objective To investigate the method to isolate and culture hepatic stellate cells ( HSCs) for studying the cellular mechanisms of hepatic frbrosis.Methods HSCs were isolated by nycodenz density gradient centrifugation after the hepatocytes obtained from adult male gebils were digested with pronase, collagenase and DNase, infused via portal vein.The cell viability was determined by trypan blue exclusion test.The purity of HSCs was identified by detectingα-SMA, desmin immunohistochemical staining.Results The yield rate of HSCs was 0.5~1 ×107 per gerbil liver, and the cell viability was more than 90%.The percentage ofα-SMA-positive cells was more than 75%after 3 days primary culture and almost 100% cells were α-SMA and desmin positive in passage culture.Conclusion The successful protocol of primary culture of Mongolian gerbil HSC provide a technical support for research of relevant liver diseases and drug development in the future.

Objective To observe the changes of learning function and neuron-specific enolase (NSE)protein of both brian and serum in rats after intermittent hypoxia,and explore the relation between NSE protein and learning and memory function.Methods Male Wistar adult rats(n=48)were randomly divided into control group (UC group) and 7.5％ chronic intermittent hypoxia group (7.5％CIH group).7.5％ chronic intermittent hypoxia model in rats were simalated by using the self-made cabin of intermittent hypoxia.At the 2nd,4th,6th,and 8th week respectively,learning and memory function in rat was tested by Morris water maze in two groups.The level of NSE protein in hippocampal CA1 region was detected by immunohistochemical method,and was detected in serum of rats by using enzyme-linked immunosorbent adsorption method.Results Compared with the control group,7.5％ CIH group at the 2nd,4th,6th,and 8th week in rats from the second week of the escape latency time was 44.13± 2.84) s,(50.35 ± 1.96) s,(57.47 ± 1.66) s,(62.85 ± 1.80) s,and across the target quadrant time was 48.81 ± 2.09) s,(42.04± 1.84) s,(36.82± 2.07) s,(31.81 ± 1.68) s.From the first two weeks,the longer the hypoxia time prolonged,the longer the rat's escapedlatency (P＜0.05) and the shorter the rats acrossed the target quadrant (P ＜0.05).7.5％ CIH group of hippocampal CA1 neurons NSE protein at the 2nd,4th,6th,and 8th week was (9.69±1.37),17.10± 1.87),(24.79± 3.51),(34.16±5.35),respectively,and serum NSE protein at the 2nd,4th,6th,and 8th week respectively was (6.03±0.91) μg/L,(11.04± 1.89) μg/L,(16.39± 1.00) μg/L,(24.22±3.73) μg/L,both of which were more than the control group.The difference was statistically significant (P＜0.05).7.5％ CIH group of hippocampal CA1 neurons and serum NSE protein expression were significant time differences,and gradually increased over time.The difference was statistically significant (P＜0.05).Positive NSE protein expression in hippocampal CA1 region relative NSE and serum protein expression values were positively correlated (r=0.94,P ＜0.01).But in control group NSE protein expression did not exist time difference (P＞0.05).Conclusion Intermittent hypoxia can cause NSE levels significant increase in serum and brain tissue,and the dynamic changes of them can reflect severity of learning and memory impairement in rat.

Objective To investigate the effect of intravenous infusion of hyper-oxygenated solution (HOS) on small intestinal ischemia-reperfusion (I/R) injury in rabbits.Methods Twenty-four rabbits of both sexes,weighing 2.5-3.2 kg,were randomly divided into 3 groups ( n =8 each):sham operation group (group S),I/R group,and HOS group.Small intestinal I/R was produced by clamping the superior mesenteric artery (SMA) for 1 h followed by 2 h of reperfusion in I/R and HOS groups,while the SMA was only clamped in group S.HOS was infused intravenously at a rate of 20 ml· kg-1 ·h -1 via the auricular vein starting from the time immediately after clamping the SMA in group HOS and the equal volume of normal saline was infused instead of HOS in group I/R.Blood samples were obtained from the inferior vena cava at 2 h of reperfusion to detect the concentration of serum lactic acid.The animals were then sacrificed and the small intestine was removed for determination of the malondialdehyde (MDA) content,and activities of superoxide dismutase (SOD),catalase (CAT) and glutathione peroxidase (GSH-Px) in intestinal tissues and for microscopic examination.The pathological changes of the intestinal epithelia were observed and the damage.to the mucous membrane was scored.The internal organs were removed and bacterial translocation from gut to the internal organs was observed.Results Compared with group S,the level of MDA and lactic acid,and rate of bacterial translocation were significantly increased,and the activities of SOD,CAT and GSH-Px were significantly decreased in groups I/R and HOS ( P ＜ 0.05).Compared with group I/R,the level of MDA and lactic acid,rate of bacterial translocation,and activities of SOD,CAT and GSH-Px were significantly decreased in group HOS ( P ＜ 0.05).Conclusion Intravenous infusion of HOS can reduce small intestinal I/R injury in rabbits.

Objective To explore the necessity and function of the sustentaculum tali screw placement for the treatment of Sanders type Ⅱ calcaneal fracture.Methods The data of Dicom that CT scan with bone was input into Mimics 12.0 software and Ansys 13.0 software for construction of calcaneus three-dimensional finite element model.Then,this model was imported into Solidworks 2010 software.Type Ⅱ calcaneal fracture model was established according to Sanders type cutting of calcaneus.Geometric parameters of AO calcaneal plate and screws were imported to the Solidworks 2010 software.Two internal fixation simulations were established based on the calcaneal model.In one model,the steel was placed on the outside with bone and the cancellous' bone screws were infiltrated respectively into the vertical bone since two screw holes which under the articular surface behind the steel plate,two screw holes which rear of the steel plate,a screw hole of the below calcaneus,two screw holes which in front of steel plate.In another model,a cortical bone screw was infiltrated into the sustentaculum tali through the bottom screw hole of articular surface of the steel plate.The two internal fixation models were loaded under the same condition,and non linear finite element analysis was carried out.The stress distribution in the two internal fixation models was calculated respectively.Results The maximum principal stress focused on the cortical bone of sustentaculum tali in both of the models under the same condition of loading.Bone joint displacement,maximum principal stress of calcaneal and internal fixation system in the model with sustentaculum screw fixation were smaller than that in the model without sustentaculum screw fixation.The stress in the model with sustentaculum screw fixation was more dispersed.Conclusion The placement of sustentaculum tali screw is essential for fixation of type Ⅱ calcaneal fracture to achieve the biomechanical stability and can be used in clinical.

OBJECTIVETo study the therapeutic method and clinical effect of domestic locking plate for treating fracture of the tubiform bone through a minimal incision and bridging fixation.

METHODSAmong 40 patients with fracture of the tubiform bone, 29 patients were male and 11 patients were female, aged from 29 to 69 years, with the average of 38.27 years. Among 34 patients with closed fracture, 7 cases were type A, 10 cases type B, 17 cases type C according to AO/ASIF classification. Among 6 patients with open fracture, 2 cases were type I, 4 cases were type II according to Gustilo classification. These patients were treated with a small incision after manipulative reduction. Locking plate and 3.5 mm diameter Kirschner wire were used to maintain the reduction and through the C-arm X-ray machine to check and adjust the position, screw were locked to bridging fixation at last.

RESULTSForty cases were followed up in 6 months on average (ranging from 4 to 10 months). The average time of union of fracture was 4.6 months (ranging from 3 to 8 months). According to Ovadia typing, the results were excellent in 16 cases, good in 20 cases, fair in 3 cases,poor in 1 case. The excellent and good rate was 90%.

CONCLUSIONSThe domestic locking plate is suitable for patients with osteoporosis and comminuted fracture because of the advantages of reliable fixation and low price. It could achieve the same effect of minimal invasive as LISS (less invasive stabilizing system) plate through the improvement of operative method.

Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation ; methods ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods

Objective To investigate the effect of ibuprofen on autophagy and astrocyte proliferation and their significances in rats with pentylenetetrazol (PTZ)-induced epilepsy. Methods Sixty male sprague-dawley rats were randomly divided into control group, PTZ group, 3-MA+PTZ group, ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group (n=12); activity of gamma-aminobutyric acid was blocked by PTZ to ignite epileptic rats in the latter 4 groups, and rats in the control group were intraperitoneally injected with normal saline once every one day; rats in the 3-MA+PTZ group and ibuprofen+PTZ group were given intraperitoneal injection of 3-MA (2.5 mg/kg) or ibuprofen (30 mg/kg) 30 min before PTZ injection;rats in the 3-MA+ibuprofen+PTZ group were given intraperitoneal injection of 3-MA (2.5 mg/kg)+ibuprofen (30 mg/kg) at the same time. The behavior and EEG features of rats were observed. Immunofluorescence staining and Western blotting were used to detect the expressions of microtubule-associated protein 1 light chain 3 (LC3) and glial fibrillary acidic protein (GFAP). Results (1) As compared with rats in the PTZ group and 3-MA+PTZ group, rats in the ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group had significantly decreased seizure grading and incidence of complete ignition, and significantly increased latency period (P<0.05). (2) The EEG waveform of the control group was normal; electroencephalogram of PTZ group and 3-MA+PTZ group showed sharp waves of high amplitude and spike waves; EEG wave peaks in the ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group decreased significantly, presenting frequent small spike waves and slow spike waves, among which ibuprofen+PTZ group showed more obvious changes. (3) The results of immunofluorescence staining and Western blotting showed that as compared with the PTZ group, 3-MA+PTZ group had statistically decreased LC3 expression and significantly increased GFAP expression (P<0.05); as compared with the PTZ group, ibuprofen+PTZ group had statistically increased LC3 expression and significantly decreased GFAP expression (P<0.05), however, 3-MA+ibuprofen+PTZ group had statistically decreased LC3 expression and significantly increased GFAP expression (P<0.05). Conclusion Ibuprofen can reduce astrocyte proliferation by promoting autophagy to affect seizures.

Objective To exlore the influence of internal quality control and external quality control assessment(EQA) resulting from applicability of control samples in measurement of whole blood viscosity (WBV) through the analysis and comparison of applicability of 1 non-Newtonian fluid internal quality control sample in 3 viscometers. Methods Viscometer B, C and D were used to measure WBV of 30 blood samples in parallel under the shear rate(SR) of 1 s-1,30 s~(-1) and 200 s~(-1), then the blood SR-WBV curves of 3 viscometers were drawn according to the results. At the same time, viscometers B, C and D were used respectively to determine the WBV of control A 10 times in one day, then the control A SR-WBV curves were mapped. Three viscometers were used to measure the manufactory control samples and control A 5 times in one day for 4 days. Four groups of daily values of manufactory control samples and control A of each instrument were used to carry out F test to calculate whether 4 daily values are difference. Finally, the control A was dispensed in 49 laboratories nationwide chosen for measurement. On the basis of viscometer used, 20 laboratories were classified as group B, 20 laboratories were classified as group C and 9 laboratories were classified as group D. Then the data under SR of 1 s~(-1) were analyzed to calculate the coefficient of variation (CV) in the group. Results There was significant difference among the WBV of blood samples measured by the viscometers B, C and D. The results under SR of 1 s~(-1) declined in turn, and they were highest under SR of 30 s~(-1) followed by the values of viscometer D and B and they were (8.14±0.75), highest under SR of 30 s-1 followed by the values of viscometer B and D, and they were (7.35±0.07), daily values of manufactory control and control A of each instruments in four groups were compared. Under SR of 1 s~(-1), there was no difference between daily values of manufactory control and control A in viscometer B (F = 2.63, 1.37, P > 0.05), and there was no difference of daily values of manufactory control among viscometer C and D (F = 0.33,3. 14, P > 0.05), but significant daily difference existed when control A was tested by viscometer C and D (F = 5.76, 8.00, P < 0.05). Under SR of 30 s~(-1), there was no difference of daily values of manufactory control among 3 viscometers(F =0.31, 0.18, 2.26, P >0.05), and there was no difference of daily values of control A among 3 viscometers' (F = 1.03, 1.83, 2.40, P > 0.05); Under SR of 200 s~(-1), there was no difference of daily values of manufactory control among 3 viscometers (F =2.59, 0.68, 2.96, P > 0.05), and there was no difference of daily values of control A among 3 viscometers (F=2.31, 3.01, 2.28, P>0.05). When control A was tested under SR of 1 s~(-1) in 49 laboratories nationwide, the WBV values in groups of viscometer B, C and D were (18.47±1.30), (11.17±2.38), viscometer D and C were 63.75% and 21.3%. Conclusions Control A could fully mimic the properties of whole blood steadily on viscometer B, but partially mimic viscometer C and D, so the control A is most appropriate for viscometer B. Because current non-Newtonian fluid internal quality control could mimic rheological properties of whole blood under specifically conditions, laboratories should evaluate the consistent degree between control and whole blood, only the candidates which can mimic the properties of whole blood approximately could be chosen as quality control of WBV. When third-party control is chosen to be samples of EQA, its applicability should be in consideration. Pretest should be performed adequately to define applicability of third-party control, so as to reduce the difference among laboratories due to applicability of control and reflect detection quality of laboratories exactly.