ObjectiveTo develop an experimental model of orthostatic hypotension (OH) after spinal cord injury (SCI) in rabbits and investigate the effect of tilting table exercise and Shengmaiyin on it. MethodsRabbit model of OH after SCI were induced by complete transection of spinal cord at the T5 spinal segment and raised to 60° head up tilt (HUT) on a tilting table. Tilting table exercise and Shengmaiyin were applied to different groups for 28 d from 4 days after operation. Systolic blood pressure (SBP) and pulse rate (PR) were observed with non-invasive sphygmobolometer of rabbit 3, 10, 17, 24, 31 d after the operation. Liquid chromatography with electrochemical detection(HPLC-ECD) was used to detect the concentrations of plasma norepinephrine (NE) and epinephrine(E) with horizontal position and HUT 60° on the 31st day. ResultsSBP with HUT 60° in rabbits of combined treatment group was significantly higher than that in rabbits of SCI group on the 24th day after operation,and the drop in SBP from horizontal position to HUT60° was significantly lower than that in rabbits of SCI group. SBP with HUT 60° in rabbits of exercise group and combined treatment group was significantly higher than that in rabbits of SCI group on the 31st day after operation. The drop in SBP from horizontal position to HUT 60° was significantly lower in rabbits of Shengmaiyin group, exercise group and combined treatment group than that in rabbits of SCI group on the 31st day after operation. The concentration of E and NE of SCI group, Shengmaiyin group, exercise group and combined treatment group was lower than that of sham group, but there was no significant difference among these groups. ConclusionThe treatment with tilting table exercise and Shengmaiyin can improve OH after SCI partially in rabbits without involving norepinephrine and epinephrine significantly. The period of treatment can be shorten when both therapies used together.

OBJECTIVETo study the therapeutic effects of posterolateral depression fractures of the tibial plateau through a modified anterolateral approach.

METHODSFrom February 2011 to January 2012,13 patients with posterolateral depression fractures of the tibial plateau were treated through a modified anterolateral approach. There were 8 males and 5 females, ranging in age from 28 to 59 years old (49.2 years old on average). Data from patients were collected retrospectively as follows: X-ray, time of fracture healing and the complications of fracture healing. The patients were evaluated both clinically and radiologically according to the Rasmussen score system.

RESULTSAll the patients were followed up, and the duration ranged from 6 to 18 months (mean 13.7 months). All the patients got bony union. The average radiographic bony union time was 15.1 weeks (ranged, 11 to 17 weeks). No case of secondary articular depression was found. No complications such as malunion or joint stiffness were found. But 1 patient had superficial infection and 1 patient had common peroneal nerve injury. According to the Rasmussen score system,the mean radiological score was 16.50 ± 0.67 (ranged, 13 to 18), and the mean functional score was 25.20 ± 2.21 (ranged, 13 to 30). The mean range of knee motion was (125.3 ± 9.3)° (ranged, 0° to 135°).

CONCLUSIONTreatment of depression fractures of posterolateral tibial plateau with a modified anterolateral approach is a safe method with effective exposure, due to its stable fixation and relatively good outcome with minimal soft-tissue complications. It is regarded as an ideal procedure for depression fractures of posterolateral tibial plateau.

Adult ; Female ; Fracture Fixation, Internal ; methods ; Fracture Healing ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tibial Fractures ; surgery

Objective:To remove murine embryonic stem cells(mESC)from the differentiating cell culture and purify the differentiated cells by Magnetic Activated Cell Sorting(MACS).Methods:Neural differentiation of mESC was induced by a 5-stage method.The specific cell surface marker,SSEA-1,was used to identify ES cells in the differentiating cells.The optimal dilutions of mouse anti mouse SSEA-1 IgM primary antibody and FITC conjugated goat anti mouse secondary antibody were determined before the flow cytometry test.The incubation time and incubation temperature of primary antibody were all optimized to make the cytometry test accurate.After the optimization,stage 4 cells were dissociated into single cell suspension,incubated with antibody of SSEA-1 and microbeads conjugated goat anti mouse IgM,and then sorted through the magnetic field.The rate of SSEA-1 positive cells in pre-and post-separation groups was assessed by flow cytometry,and the viability of cells was evaluated by trypan blue staining counting under light microscopy.Results:The proportion of SSEA-1 positive cells in the separated cells can be reduced from(7.19?1.36)% to(1.34?0.80)%.The survival rate of sorted cells was more than 92%,similar to that of pre-separation cells.Conclusions:The MACS system we used can effectively remove mESC from the differentiated cells.The sorted cells will be well provided for the subsequent studies about transplantation therapy.

Adult ; Humans ; Male ; Nasal Obstruction ; congenital ; surgery ; Nose ; abnormalities

Objective To construct single chain antibody specific to membrane protein of Schistosoma japonicum by gonetic engineering technique. Methods The V\-H (heavy-chain variable region) and V\-L (light-chain variable region) genes were amplified by PCR from the genomic DNA of NP11-4 cell line, and sequenced by Sanger's method. The ScFv was constructed in pTHA90 vector using V\-H and V\-L genes, then expressed by IPTG. Results The V\-H and V\-L genes were obtained through PCR. The DNA sequences showed that V\-H and V\-L were new variable region genes of antibody. They were registered by GenBank. A ScFv gene with (Gly4Ser) 3 intralinker in the pTHA90 vector was successfully constructed. The ScFv was expressed as thioredoxin-fused proteins about 36\^2 kDa. Conclusion A specific ScFv against the membrane protein of Schistosoma japonicum was constructed and expressed.

Objective To assess the expression and significance of proprotein convertase subtilisin kexin 9 (PCSK9) in patients with rheumatoid arthritis (RA).Methods Sixty-five RA patients and forty-seven healthy controls were recruited in this study.The body mass index (BMI) and serum total cholesterol (TC),triglyceride (TG),high density lipoprotein (HDL),lipoprotein a,low density lipoprotein (LDL),very low density lipoprotein(VLDL),apolipoprotein A (ApoA),apolipoprotein B (ApoB) and the ratio of LDL-C/HDL-C were tested.Other parameters included disease activity score 28 (DAS28),rheumatoid factor (RF),anti-cyclic citrullinated peptide (CCP) antibody,erythrocyte sedimentation rate (ESR),c-reactive protein (CRP).Serum PCSK9 level was measured by ELISA and compared between RA patients and healthy controls.Results (1) The serum PCSK9 levels in RA patients were higher than those in healthy controls [(409.36 ±223.52) μg/L vs (292.19 ± 109.79) μg/L,P ＜ 0.05].(2) Compared with subgroup of moderate and low active disease and patients in remission,PCSK9 was significantly higher in patients with highly active disease (P ＜ 0.05).(3) The serum PCSK9 levels were positively correlated with RF,TC,TG,LDL,very low density lipoprotein (VLDL),ApoB,with r values as 0.303,0.490,0.320,0.451,0.319,0.463,respectively (P ＜ 0.05).(4) Multiple stepwise regression analysis showed that DAS28,RF,TC and LDL-C/HDL-C were relevant factors for PCSK9 in RA patients.Conclusions The serum PCSK9 level is elevated in RA patients,which is related to RF,disease activity,TC,TG,LDL,VLDL,ApoB.This suggests that PCSK9 is potentially linked to inflammatory reaction and lipid metabolism in rheumatoid arthritis.

Objective:To develop evaluation index system of Knowledge, Attitude, and Practice Model of esophageal speech training for Otolaryngology nurse, which can provide an initial tool to evaluate the effect of esophageal speech training for Otolaryngology nurse.Methods:Based on the Knowledge, Attitude, and Practice Model and literature research, the initial dimensions and items were developed. The evaluation index system was completed by Delphi technique that 23 experts were invited to evaluate the importance of the dimensions and items in the consulting round. Then the weight value of the index system was computed through analytic hierarchy.Results:For the first and second rounds of the study, respectively, the recovery rates were 78.26% and 100.00%, the expert authority were 0.85 and 0.86, and the Kendall′s W were 0.16 and 0.19 ( P<0.05). Finally, the index system contained 3 dimensions and 30 items. Conclusions:The evaluation index system has the possible scientificity and reliability, and has the feasible thoery and content. In the future, the evaluation index system can be used to evaluate the effect of esophageal speech training for Otolaryngology nurse through the clinical practice improvement.

OBJECTIVENeonatal hypoxic-ischemic encephalopathy (HIE) harms the lives and health of newborn infants and children severely. The prognosis is not satisfied, especially of the severe HIE. Mesenchymal stem cells (MSCs) can secrete a series of growth factors and neurotrophic factors. As well they have the potential ability to differentiate to the neural cells in vitro and in vivo. Therefore MSCs transplantation has been employed as a source of progenitor cells for cell therapy in patients with HIE in order to promote recovery of brain function and reduce the sequelae. Studies have shown that MSCs could enter the cerebral parenchyma and differentiate to neural cells through systemic infusion, but most of the researches applied adult stroke animal models. This study used neonatal HIE models to test the hypothesis that MSCs could enter the brain of newborn Wistar rats through the blood-brain barrier (BBB) by intraperitoneal infusion followed by observing the characteristics of the distribution and differentiation of MSCs in brain tissues, and exploring the effects of hypoxic-ischemic brain damage to the penetration and differentiation of MSCs.

METHODSIsolation and purification of MSCs were established from the whole bone marrow of juvenile Wistar rats by removing the nonadherent cells in primary and passage cultures. For cellular identification, MSCs of three to five passages were continuously pre-labeled with 5-bromo-2-deoxyuridine (BrdU) for 72 hours before transplantation. Animal models of HIE were built in 7-day-postnatal Wistar rats according to the method described by Rice. Two hours after hypoxia-ischemia, rats in HIE group (n = 8) were intraperitoneally infused with MSCs (4 x 10(6), 0.5 ml). In control group (n = 8), 7-day-postnatal normal Wistar rats were intraperitoneally infused with the same amount of MSCs. All rats were sacrificed and their cerebra were sectioned by cryomicrotome 14 days after transplantation. Immunohistochemical staining with chromogen diaminobenzidine (DAB) was used to detect and measure the cells derived from MSCs, and study the characteristics of distribution. To determine the differentiation of the BrdU positive cells entering the brains, immunofluorescence double labeling for BrdU and neural cells specific antigens was performed.

RESULTSMSCs were distributed throughout the cerebra in both groups at the 14th day after transplantation. The number of MSCs detected was 2415 +/- 226 in the control group, and 3626 +/- 461 in HIE group, respectively (t = 6.68, P < 0.05). More BrdU reactive cells were observed in the right ischemic hemisphere (1904 +/- 267) than in the contralateral hemisphere (1723 +/- 204), (t = 4.47, P < 0.05). No significant difference was found while comparing both cerebral hemispheres of the control group (t = 0.31, P > 0.05). In the HIE group, MSCs distributed more extensively, and some focal aggregations of MSCs were noticed. A few MSCs expressed Nestin-protein marker of neural progenitor cells, and almost none of the MSCs which expressed proteins characteristic of neuron (e.g. NSE) and astrocyte (e.g. GFAP) was detected at the 14th day after transplantation.

CONCLUSION1. MSCs could enter the cerebral parenchyma through BBB and migrate throughout the brain by intraperitoneal infusion. 2. More MSCs injected intraperitoneally were localized and directed to the sites of hypoxic-ischemic brain damage. 3. Transplanted MSCs could not differentiate to neuron and astrocyte without other interventions during 14 days after transplantation.

Animals ; Blood-Brain Barrier ; physiology ; Brain ; Cell Differentiation ; Cell Movement ; Disease Models, Animal ; Hypoxia-Ischemia, Brain ; therapy ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; physiology ; Rats ; Rats, Wistar

OBJECTIVETo test the reliability of quantitative neurological tests in evaluating the neurological deficits in rat models of cerebral ischemia.

METHODSForty male SD rats (7 to 9 weeks old) were subjected to transient (1 h) middle cerebral artery occlusion (MCAO) to induce cerebral infarction and 9 received sham operation only. The motor and sensory functions of the rats were measured before and 1, 3, 7 and 14 days after MCAO by accelerating rotarod test and modified adhesive removal test. Fourteen days later, the rats were sacrificed for histological examination, and the lesion size was determined by Winroof image analysis. The neurological deficits of the rats with different lesion sizes and at different time points were analyzed.

RESULTSAccording to the infarct size, the rats after MCAO were divided into two subgroups with large lesions [(68.56+/-12.54)%] affecting both the cortical area and basal ganglia and small lesions [(12.45+/-9.06)%] affecting the basal ganglia. Significant differences in the results of modified adhesive removal test and rotarod test were found between the two groups, and a liner relationship was noted between the lesion size and the results of the neurological tests.

CONCLUSIONThe modified quantitative neurological tests can reliably evaluate the severity of the infarction and estimate the infarct size in rats with cerebral infarction.

Animals ; Behavior, Animal ; physiology ; Brain Ischemia ; pathology ; physiopathology ; Infarction, Middle Cerebral Artery ; pathology ; physiopathology ; Male ; Motor Activity ; Neurologic Examination ; Psychomotor Performance ; Rats ; Rats, Sprague-Dawley ; Severity of Illness Index

Objective To investigate the expression of peroxisome proliferator-activated receptor (PPAR?)in lupus nephritis(LN)patients and the possible mechanisms of PPAR?in the pathogenesis of LN. Method PPAR?expression was examined in 21 LN patients and 5 normal kidney biopsy specimens by im- munohistochemical method.The relationship between PPAR?expression and renal pathologic changes was an- alyzed.Results Glomerular and tubular positive staining of PPAR?in LN patients was markedly up-regulated compared with that in normal kidney specimens.The distribution and expression of PPAR?in classⅣwas sig- nificantly increased compared with that in classⅤandⅡ.The relevance assay showed that there was positive relationship between active index and glomerular PPAR?immunohistochemistry staining cell numbers(r=0.94, P＜0.01 ).Conclusion This study demonstrates in vivo that PPAR?expression is increased in active LN pa- tients with pathological active inflammation.These data suggest that the increase of PPAR?expression in renal cells may play an important role in the pathogenesis of LN.