Dynamic elastic modulus assessment of the early degeneration model of an intervertebral disc in cynomolgus monkeys with one strike loading.

OBJECTIVE: Disc degeneration has long been associated with excessive mechanical loading or acute disc injury. Our goal is to perform a shock load on the functional units of the cynomolgus monkey intervertebral disc and analyze the degree of degeneration of the intervertebral disc through image analysis and comprehensive analysis. The organ model establishes a standard organ culture model and a non-invasive biomechanical evaluation protocol close to the early degeneration of the human intervertebral disc. METHODS: After modeling, the cynomolgus monkey intervertebral discs were collected and loaded into the dynamic mechanical culture system. The physiological group was loaded with 10% high compressive deformation load for one second, the injury group was punctured with annulus fibrosus, the model group was loaded with 20-50% high compressive deformation, and the nutritional components were a high-glucose group and low-glucose group. After day 3 (short term) and day 10 (long term), samples were collected to analyze cell viability, histomorphology, image analysis for imaging and biomechanical changes. RESULTS: Both the injury group and the 30-50% strain model group showed signs of early degeneration, including decreased instantaneous compressive stiffness, percent change in gray value, decreased cell viability, AF fissure, and percent increase in dynamic elastic modulus. The glucose-restricted group also showed signs of early disc degeneration in long-term cultures. CONCLUSION: This study shows that a single shock load can induce early degeneration of healthy cynomolgus monkey intervertebral discs, and 30% strain may be the nociceptive threshold for early degeneration of healthy intervertebral discs. More importantly, a non-invasive biomechanical evaluation scheme of Percentage change in dynamic modulus of elasticity is established, which solves the key scientific problem of how to non-invasively, quantitatively and sensitively detect the development process of early intervertebral disc degeneration and its degree of degeneration in an in vitro organ model.

Programmed NP Cell Death Induced by Mitochondrial ROS in a One-Strike Loading Disc Degeneration Organ Culture Model.

Increasing evidence has indicated that mitochondrial reactive oxygen species (ROS) play critical roles in mechanical stress-induced lumbar degenerative disc disease (DDD). However, the detailed underlying pathological mechanism needs further investigation. In this study, we utilized a one-strike loading disc degeneration organ culture model to explore the responses of intervertebral discs (IVDs) to mechanical stress. IVDs were subjected to a strain of 40% of the disc height for one second and then cultured under physiological loading. Mitoquinone mesylate (MitoQ) or other inhibitors were injected into the IVDs. IVDs subjected to only physiological loading culture were used as controls. Mitochondrial membrane potential was significantly depressed immediately after mechanical stress (P < 0.01). The percentage of ROS-positive cells significantly increased in the first 12 hours after mechanical stress and then declined to a low level by 48 hours. Pretreatment with MitoQ or rotenone significantly decreased the proportion of ROS-positive cells (P < 0.01). Nucleus pulposus (NP) cell viability was sharply reduced at 12 hours after mechanical stress and reached a stable status by 48 hours. While the levels of necroptosis- and apoptosis-related markers were significantly increased at 12 hours after mechanical stress, no significant changes were observed at day 7. Pretreatment with MitoQ increased NP cell viability and alleviated the marker changes by 12 hours after mechanical stress. Elevated mitochondrial ROS levels were also related to extracellular matrix (ECM) degeneration signs, including catabolic marker upregulation, anabolic marker downregulation, increased glycosaminoglycan (GAG) loss, IVD dynamic compressive stiffness reduction, and morphological degradation changes at the early time points after mechanical stress. Pretreatment with MitoQ alleviated some of these degenerative changes by 12 hours after mechanical stress. These changes were eliminated by day 7. Taken together, our findings demonstrate that mitochondrial ROS act as important regulators of programmed NP cell death and ECM degeneration in IVDs at early time points after mechanical stress.

Local Infiltration Analgesia with Ropivacaine Improves Postoperative Pain Control in Ankle Fracture Patients: A Retrospective Cohort Study.

Purpose: The study aimed at investigating the effect of local infiltration analgesia (LIA) with ropivacaine on postoperative analgesia for patients undergoing ankle fracture surgery. Methods: Consecutive patients were retrospectively included and analysed according to their medical records from July 2014 to August 2018 in a tertiary hospital. Inclusion criteria were patients undergoing open reduction and internal fixation (ORIF) for ankle fractures under general anaesthesia. Moreover, patients should have received intravenous patient-controlled analgesia (iPCA) or LIA + iPCA for postoperative pain relief. The primary outcome indicator was visual analogue scale (VAS) from 8 hours to 48 hours after surgery. Secondary outcomes included postoperative opioid requirement, need for rescue medication, opioid-related adverse effects, and wound complications. Results: In total, 89 consecutive patients were included in the study. There were 48 males and 41 females. The average age was 44.6 ± 7.0 years, and VAS scores were significantly lower in the LIA + iPCA group at 8 hours after surgery (1.51 ± 0.58 cm vs 4.77 ± 1.83 cm, p < 0.001). The time to first tramadol consumption was longer (580 ± 60.9 minutes vs 281 ± 86.4 minutes, p < 0.001). The time to first tramadol consumption was longer (580 ± 60.9 minutes vs 281 ± 86.4 minutes, p < 0.001). The time to first tramadol consumption was longer (580 ± 60.9 minutes vs 281 ± 86.4 minutes, p < 0.001). The time to first tramadol consumption was longer (580 ± 60.9 minutes vs 281 ± 86.4 minutes, p < 0.001). The time to first tramadol consumption was longer (580 ± 60.9 minutes vs 281 ± 86.4 minutes. Conclusions: The retrospective cohort study indicates that LIA with ropivacaine can provide better early postoperative pain management with a reduction of VAS scores for ankle fracture surgery. Patients receiving wound infiltration also experience decreased opioid consumption, a lower rate of analgesia-related side effects, and comparable wound complication rate.

Efficacy of open reduction and internal fixation with a miniplate and hollow screw in the treatment of Lisfranc injury.

PURPOSE: To investigate efficacy of open reduction and internal fixation with the miniplate and hollow screw in the treatment of Lisfranc injury. METHODS: Ten cases of Lisfranc injury treated by open reduction, miniplate and hollow screw in our hospital were retrospectively analyzed. There were 6 males and 4 females with age ranging from 25 to 45 years (mean 32 years). Among them, one case was classified as Type A, six Type B and three Type C. Injury mechanism included road traffic accidents (3 cases), fall from height (5 cases) and hit by heavy object (2 cases). All injuries were closed without cerebral trauma or other complicated injuries. The time interval between injury and operation was 6-10 days (average 6.6 days). Postoperatively, the foot function was assessed using Visual Analogue Scales (VAS) and American Orthopaedic Foot and Ankle Society (AOFAS) Scales. Healing time and complications were observed. RESULTS: All patients were followed up for 18-24 months (average 20 months). Anatomic reduction was achieved in all patients on images. There was statistical significance between preoperative score (7.89 ± 0.34) and score at postoperative 8 weeks (0.67 ± 0.13). According to the AOFAS score, 5 cases were defined as excellent, 3 cases as good and 2 cases as fair. During follow-up, there was no wound infection or complications except for osteoarthritis in 2 cases. Healing time ranged from 3 to 6 months with an average of 3.6 months. CONCLUSION: Anatomical reduction of Lisfranc injury can be achieved by open reduction and internal fixation with the miniplate and hollow screw. Normal structure of Lisfranc joint is regained to a great extent; injured ligaments were also repaired. Therefore, this method offers excellent curative effect and can avoid postoperative complications and improve the patients' quality of life.

[Non-invasive predictive model of esophageal varices in cirrhosis].

OBJECTIVE: To analyze the non-invasive indexes for predicting esophageal varices (EV) in liver cirrhosis, and to establish a model for predicting the degree of EV. METHODS: A total of 294 patients with liver cirrhosis and portal hypertension were divided into the following groups according to EV grade as assessed by endoscopy: non-EV and grade I EV, grade II EV and grade III EV. The non-invasive EV predictive measures of liver stiffness (LS), platelet (PLT) count, spleen thickness (ST), PLT/ST ratio, portal vein diameter, portal vein flow velocity and Child-Pugh score (CPS) were assessed by univariate analysis and multivariate logistic regression analysis, and used to generate a predictive model. The t-test, chi-square test, logistic analysis and receiver operating characteristic (ROC) curve were used in statistical analyses. RESULTS: The area under the ROC for the new model was 0.990. The best cutoff value for the score was 0.898, as defined from the ROC. The sensitivity of the model was 96.5%, and the specificity was 99.2%. CONCLUSIONS: The model for predicting EV was composed of LS, PLT count, ST, PLT/ST and CPS, which was accurate and sensitive, and could be used to predict EV in clinic.

[Bottleneck and development trend of bone xenograft for the treatment of bone defect].

Bone xenograft bone for the treatment of bone defect is one of the current research focus, which has advantages of extensive sources, low cost, simple preparation method. While the process of single bone xenograft bone in repairing bone defect is very long, and the clinical outcome is not satisfactory. The main problems focus on formation of bone and vascularization. Reconstituted bone xenograft combined with cells and xenogenic bone material could promote vascularization and bone fusion in vivo, thus achieve a clinical effect of autogenous bone in repairing bone defect.

Evaluation of partially purified soluble egg antigens in colloidal gold immunochromatography assay card for rapid detection of anti-Schistosoma japonicum antibodies.

Shistosomiasis is one of the important parasitic diseases in developing countries and especially remains a threat to public health in China. Many immunodiagnostic kits have shown cross-reactions with other parasitic diseases and need large volumes of serum for the tests. In this study, we evaluated partially purified soluble egg antigen (SEA) in a colloidal gold immunochromatography assay (GICA) card kit for rapid detection of anti-Schistosoma japonicum antibodies using 5 microl of serum. Partially purified SEA from S. japonica was purified by Sephacryl S-300 chromatography. The optional reaction system and detection level of GICA using partially purified SEA were established by improving conjugated concentration and formulation of sample buffer and labeled solution. GICA showed 93.7% sensitivity in detecting schistosomiasis patients, 97.6% specificity in healthy population and patients with other parasitic diseases and a Youden's index value of 0.91. Cross-reaction with other parasitic diseases, such as paragonimiasis (1 case) and toxoplasmosis (1 case) is significantly lower compared to using crude SEA. Partially purified SEA in GICA is practical for detection of schistosomiasis in the field as it requires a small volume of serum, has high sensitivity, and has low cross-reaction rate.

Prognostic value of CYP2W1 expression in patients with human hepatocellular carcinoma.

CYP2W1 overexpression has been reported in a variety of human cancers. However, the role of CYP2W1 in hepatocellular carcinoma (HCC) remains unclear. This study was designed to evaluate the expression and prognostic significance of CYP2W1 in human HCC. Real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was conducted to detect CYP2W1 messenger RNA (mRNA) expression in 41 pairs of fresh-frozen HCC tissues and adjacent noncancerous tissues. In addition, CYP2W1 expression was analyzed by immunohistochemistry in 133 clinicopathologically characterized HCC cases. The relationship between CYP2W1 expression and clinicopathological features was analyzed by appropriate statistics. Kaplan-Meier analysis and Cox proportional hazards regression models were used to investigate the correlation between CYP2W1 expression and prognosis of HCC patients. The relative mRNA expression of CYP2W1 was significantly higher in HCC tissues than in adjacent noncancerous tissues (P < 0.001). In addition, CYP2W1 expression was significantly correlated with tumor size (P = 0.023), histological differentiation (P = 0.04), and tumor stage (P = 0.014). The Kaplan-Meier survival curves indicated that patients with high expression of CYP2W1 had shorter overall survival than those with low expression (P < 0.001). Furthermore, Cox regression analyses showed that CYP2W1 expression was an independent predictor of overall survival. Our data suggest that CYP2W1 could play an important role in HCC and might serve as a valuable prognostic marker and potential target for gene therapy in the treatment of HCC.

[Changes of erythrocyte rheology in rabbits with acute renal failure].

OBJECTIVE: To observe the change of erythrocyte theology in rabbits with acute renal failure (ARF). METHODS: Thirty-eight healthy rabbits were randomly divided into control group (n = 8), model group (establishing ARF model via intramuscular injection 1% HgCl2, and divided into 12 h, 24 h, 48 h subgroups, all n = 10), the arterial blood sample was taken out through carotid artery at corresponding times after anesthetization with urethane, for detecting the indices of renal function and erythrocyte rheology. RESULTS: The levels of urea and creatinine in plasma of model rabbits at 12 h, 24 h and 48 h were higher than those of control group, and there was a rise trend along with the time extension. The erythrocytes electrophoresis time at 12 h of model group was higher, the electrophoresis rate and migration rate of erythrocytes were lower compared with those of control group; the erythrocytes electrophoresis time at 24 h of model group was lower and the electrophoresis rate and migration rate were higher compared with those of model group at 12 h; and there were no statistical differences in erythrocytes electrophoresis indices between model group at 48 h and other groups. Meanwhile, there was a rise trend in erythrocyte sedimentation rate (ESR), K value of equation and emendation along with the time extension of ARF, but these indices only at 48 h of model group was lower significantly than that of control group. There were no statistical differences in aggregation index and deformability index of erythrocytes among groups. CONCLUSION: During the process of ARF, the erythrocytes electrophoresis time lengthen and electrophoresis rate and migration rate decrease at early stage, and these indices gradually return to normal; the indices of ESR increase gradually.

[Development and evaluation of colloidal gold immunochromatography assay (GICA) for detection of antibody against Schistosoma japonica in smaller serum].

OBJECTIVE: To develop a rapid colloidal gold immunochromatography assay (GICA) for detection of the antibody to schistosome in 5 microl sera. METHODS: A soluble egg antigen (SEA) from Schistosomajaponica was separated with sephacryl S-300 column to retain the active antigen fractions to the antibody of schistosome. The optional reaction system and detection kit with 5 microl sera were established by improving conjugated concentration and formulas of sample buffer and labeled solution. RESULTS: The sensitivity of detecting schistosomiasis patients whose stool examinations for schistosome eggs were positive was 93.7%, the specificity to health population 97.1%, the cross reaction rate to patients with paragonimiasis 5.6%. The Youden' s index value was 0.91. There was 96.1% crude coincidence of GICA and ELISA in detecting 507 cases of floating population and the Kappa value was 0.81. CONCLUSION: GICA kit is practical for detection of schistosomiasis in the field because of its advantages such as smaller serum needed, the high sensitivity, lower cross reaction rate and spread application for human and animals.

Effect of serum from sub-healthy subjects with fatigue on the gene expression profile of skeletal muscle cells.

To investigate the effect of serum from sub-healthy subjects with fatigue on the gene expression profile of skeletal muscle cells (HSkMCs), HSkMCs were cultured in vitro and treated with serum from healthy and sub-healthy subjects independently. Total RNA was extracted, purified and used for reverse transcription into cDNA. This was labeled with one of two fluorescence dyes, Cy3 or Cy5, and hybridized with a Human Genome Array carrying 8064 target genes. The differentially expressed genes were selected with the aid of software. There were 86 differentially expressed genes identified between the sub-healthy subjects and the healthy individuals, of which 60 were upregulated and 26 were downregulated compared with those of the healthy individuals. These genes were mainly associated with the cell cycle, membrane channels, protein transport, energy metabolism and apoptosis. Our results suggest that serum from sub-healthy subjects with fatigue may change the gene expression profile of HSkMCs.

Changes of the tubular markers in type 2 diabetes mellitus with glomerular hyperfiltration.

AIM: To assess whether glomerular hyperfiltration (GHF) could result in renal tubular damage in type 2 diabetes mellitus (T2DM) patients. METHODS: Reference value of estimated glomerular filtration rate (eGFR) was determined in 248 healthy individuals based on serum CysC levels. GHF was defined as an eGFR exceeding the sex-specific 97.5th percentile in non-diabetic individuals. In the present study, 30 with GHF, 58 with norm-GFR T2DM, and 24 healthy controls were recruited. Tubular markers, such as urinary N-acetyl-ß-D-glucosaminidase (NAG) and kidney injury molecule 1 (KIM-1), as well as serum and urinary neutrophil gelatinase-associated lipocalin (NGAL), were measured and compared. The correlation of these markers with eGFR was analyzed in the GHF group. RESULTS: The GHF group had higher urinary NGAL and KIM-1 levels but lower serum NGAL level than the norm-GFR and control groups. Slightly decreased serum NGAL and increased urinary NGAL levels were also noted in the norm-GFR group compared with those of the controls. There was no statistical difference in the urinary NAG values among the three groups. Correlation analysis showed that eGFR was positively related to fasting blood glucose (FBG), HbA1c, urinary NGAL, and KIM-1, but negatively with serum NGAL in the GHF group. CONCLUSION: Higher urinary tubular damage markers were found in T2DM patients with GHF than the norm-GFR and control groups, probably a direct proof that GHF is a deleterious factor for diabetic nephropathy.

[Mechanism of multiple organ injury subsequent to acute renal failure with respect to membrane pump activity in rabbits].

OBJECTIVE: To observe the changes in membrane pump activity of kidney, myocardium and pancreas in rabbits with acute renal failure (ARF) in rabbits, and inquire into the mechanism of multiple organ injury subsequent to ARF. METHODS: Forty-two rabbits were randomly divided into control group, HgCl(2) group and glycerine group, and the latter two groups were subdivided into 12, 24, 48-hour subgroups, with 6 rabbits in each group. The ARF model was reproduced by hypodermic injection 1% HgCl(2) (1.3 ml/kg) in HgCl(2) group, or intramuscularly injection 50% glycerine (10 ml/kg) in glycerine group, respectively. At different time points, the kidney, myocardium and pancreas were harvested, and homogenates of them were prepared. The ATPase activities of different organ homogenates were determined. RESULTS: It showed that the activities of Na(+)-K(+)-ATPase, Ca(2+)-ATPase, Ca(2+)-Mg(2+)-ATPase of renal homogenate in two model groups were reduced gradually with worsening of renal function, and they became lowest at 48 hours [(0.84±0.16), (0.52±0.17), (0.45±0.09) µmol×mg(-1)×h(-1) in HgCl(2) group; (0.85±0.22), (0.49±0.21), (0.54±0.17) µmol×mg(-1) ×h(-1)in glycerine group]. The respective activities of Na (+)-K (+)-ATPase, Ca (2+)-ATPase, Mg(2+)-ATPase, Ca(2+)-Mg(2+)-ATPase of myocardium and pancreas homogenates in two model groups were reduced gradually following depression of renal function, and they became lowest at 48 hours [(0.56±0.11), (0.51±0.19), (0.55±0.19), (0.37±0.19) µmol×mg(-1)×h(-1) in HgCl(2) group and (0.52±0.19), (0.62±0.10), (0.61±0.16), (0.54±0.10) µmol×mg(-1) ×h(-1) in myocardium homogenate of glycerine group; (0.81±0.12), (0.71±0.15), (0.73±0.18), (0.62±0.16) µmol×mg(-1) ×h(-1) in HgCl(2) group and (0.72±0.13), (0.57±0.18), (0.66±0.14), (0.59±0.23) µmol×mg(-1) ×h(-1) in pancreas homogenate of glycerine group], there was statistical difference compared with control group (P<0.05 or P<0.01). CONCLUSION: The mechanism of myocardial and pancreatic injury subsequent to ARF might be related to reduction of the activity of cell membrane pump.

The mechanism of myocardium and pancreas injury in rabbits with acute renal failure might be related to myeloperoxidase and membrane pump activities.

There is increasing evidence indicating that the distant organ injury is a major contributor of high mortality in patients subjected to acute renal failure (ARF). However, sources and mechanisms that ARF causes distant organ injury remain to be determined. The aim of this study is to explore the mechanism from polymorphonuclear neutrophil (PMN) sequestration and membrane pump suppression. To achieve this, we examined myeloperoxidase (MPO), a marker of PMN accumulation in tissues, and membrane pump activities of heart, pancreas, and kidney in two ARF rabbit models. Rabbits are randomly assigned to control, HgCl(2)-treated, and glycerin-treated groups. ARF animal models are established by hypodermic injection of 1% HgCl(2) with 1.3 mL/kg bodyweight (bw) in HgCl(2)-treated group or intramuscular injection of 50% glycerin with 10 mL/kg bw in glycerin-treated group, respectively, and all animals in each group are further divided into 12 h, 24 h, and 48 h subgroups with each consisting of six rabbits. Six healthy rabbits serve as control group. Results have shown that MPO activities of kidney, myocardium, and pancreas in two model groups were significantly increased than control group at diverse time points. Membrane pump activities of kidney in two model groups are significantly lower than the control group at multiple time points. Moreover, Na(+)-K(+)-, Ca(2+)-, Mg(2+)-, and Ca(2+)-Mg(2+)-ATPase activities of myocardium and pancreas in two model groups are gradually declined with the development of ARF. These findings suggest that PMN sequestration and membrane pump suppression plays an important role in the pathogenesis of ARF and also a major mechanism of myocardium and pancreas injury during the process of ARF.

[Retrograde intramedullary locked nail and bolt for comminuted fracture in the femur condyles].

OBJECTIVE: To evaluate the therapeutic effects of locked supracondylar intramedullary nail and bolt for treating comminuted fracture in the distal end of the femur. METHODS: Forty-two patients (including 31 male and 11 female patients) with supracondylar and intercondylar fractures, classified into type C1 in 20 cases and type C2 in 22 cases according to AO classification, were treated with retrograde intramedullary nail and bolt. RESULTS: The follow-up period ranged from 3 to 18 months and the average time of bone healing was 4.6 months. Assessed by functional scoring, the results were excellent in 31 patients, good in 10, and acceptable in 1, with the rate of excellent result as high as 97.6%. CONCLUSIONS: Retrograde intramedullary nail and bolt can be ideal for treating supracondylar and intercondylar fractures. The bolt can effectively control the dissociation of the intercondylar fractures and good functional recovery of the knee joint can be achieved through early functional training.