Volume 93  ·   Issue 17  ·  2013

To establish the experimental autoimmune Lewis rat model of myasthenia gravis (EAMG) specific for muscle-specific kinase (MuSK).

Lewis rats were immunized subcutaneously with a single injection of 200μl of MuSK (50/100μg) emulsified in complete Freund′s adjuvant (CFA).And MuSK was expressed according to the murine gene of MuSK (AY360453).Control rats were immunized with CFA alone.All rats were observed every other day in a blinded fashion for fatigue characteristics.The clinical symptoms were graded between 0 and 3.And the clinical scores were recorded.The serum levels of anti-MuSK IgG at Days 7 and 57 were determined by enzyme-linked immunosorbent assay (ELISA).

The clinical symptoms of myasthenia garvis (MG) appeared at Day 16 and peaked at Days 18- 22.The serum levels of anti-MuSK IgG were markedly elevated in the experimental groups compared with the control group.

EAMG induced by MuSK is successfully established in rats.And this animal model may help us elucidate the pathogenesis of anti-MuSK myasthenia (AMM) and discover new drug therapy in the future.

To explore the correlations of serum total bilirubin (TBIL), direct bilirubin (DBIL), indirect bilirubin (IBIL) and uric acid (UA) with myasthenia gravis (MG).

A total of 131 MG patients were selected as MG group and 176 healthy cases as control group.They were enrolled from the Department of Neurology, First Affiliated Hospital, Anhui Medical University between January 2010 and December 2012.And the controls were recruited from a health check-up center.The total serum BIL and UA concentrations were measured by an enzymatic method.All MG patients underwent thymus computed tomography (CT) scanning.

The serum levels of TBIL, IBIL and UA in patients with MG ((11.0±0.4, 7.4±0.3, 270±70) μmol/L) were significantly lower than those in healthy control group ((13.0±0.4, 9.6±0.3, 301±60) μmol/L)(P<0.01).Moreover, these results were consistent when the male and female cohorts were investigated separately.In MG group, females had significantly lower serum TBIL, IBIL and UA levels ( (10.1±3.7, 6.7±2.7, 242±68) μmol/L) than males ((12.2±5.5,8.4±4.2,288±73) μmol/L)(P<0.05-0.01).However, no difference existed when comparing different grades of MG patients according to the modified Osserman classification.Also no significant difference existed between MG patients with thymic abnormalities and those without.In comparison with the subjects in the reference group (total bilirubin≥11.4μmol/L, UA≥279μmol/L), the odds ratio (95% CI) for MG patients in the lower tertile (total bilirubin<11.4μmol/L, UA<279μmol/L) were 1.98 (1.20-3.29) and 2.22 (1.29-3.82) respectively after multivariable adjustment.Serum level of TBIL and UA had positive correlations with creatintine as indicated by Pearson correlation analysis (r=0.151, P=0.008|r=0.301, P=0.000).Meanwhile,serum level of UA had a negative correlation with high density lipoprotein-cholesterol (r=-0.347, P=0.000).

Decreased serum levels of BIL and UA are closely correlated with MG.As a replacement therapy, administration of BIL and UA or theirs precursors may offer benefits to the MG patients.

To explore the characteristics of acute exacerbations of myasthenia gravis after fluoroquinolone exposure.

Gender, age, prior type, absolute score, concurrent disease, precipitated disease, use of antibiotic, onset/symptom/degree of exacerbation, therapeutic measures and prognosis at Month 1 were retrospectively analyzed for 9 patients after fluoroquinolone systemic exposure.Results Ciprofloxacin (n=4), levofloxacin (n=1) and moxifloxacin (n=4) exposure resulted in myasthenia gravis exacerbation.Myasthenia gravis exacerbations developed at 15 minutes to 4 days post-exposure.And the clinical scores of quantitative myasthenia gravis (QMG) increased by an average of 10.The main syndromes included dyspnea, diplopia, ptosis and dysphagia.All patients improved upon the withdrawal of fluoroquinolone in conjunctions with other interventions.

Fluoroquinolone exposure may result in myasthenia gravis exacerbations in patients with underlying diseases.Healthcare professionals should be aware of this serious drug-disease association.

crush syndrome refers to a clinical syndrome characterized by limb swelling, necrosis, hyperkalemia, myoglobinuria and acute kidney injury when the muscle-rich parts of the limbs or trunk are squeezed for a long time. The core link of squeeze syndrome is rhabdomyolysis, which causes the contents of muscle cells to leak into extracellular fluid and blood circulation, resulting in a series of complications such as reduced effective circulating blood volume, electrolyte disorder, acute kidney injury and multiple organ dysfunction. Earthquakes, landslides, mine collapses, mudslides, hurricanes, wars, etc. may produce a large number of patients with crush syndrome^[1,2,3,4,5]。

To explore the diagnostic rationales for pelvic retroperitoneal tumors and summarize their therapeutic regimens.

A total of 16 retroperitoneal tumor patients were recruited.And their general information, previous medical history, physical examinations, auxiliary tests, surgical findings and postoperative pathological results were analyzed.

Two cases were diagnosed through preoperative magnetic resonance imaging (MRI) while others found intraoperatively.Complete tumor resection was performed in all except for one case.Postoperative pathological examinations revealed 10 benign cases.And there was one case of pelvic endometriosis (mild cytologic atypia).Five cases were malignant.The operation duration was 1.45-8.5 hours and peri-operative bleeding volume 50-5000 ml.Among them, 12 patients had heavy adhesion in pelvic cavity, 7 cases underwent operations collaboratively with related departments because of surgical difficulties and vascular injury and bladder rupture occurred in 1 case.During the follow-ups,one case was lost, two patients died from disease recurrence and another one had a postoperative relapse at Month 4.The other 12 cases recovered well and had no recurrence.

Surgery remains a key for retroperitoneal tumors.With a low diagnostic rate, they are often found surprisingly intraoperatively.Because of surgical difficulties and frequent complications, multi-departmental collaboration is necessary.Preoperative correct diagnosis and adequate preoperative preparation are essential.And MRI is an effective auxiliary examination.

To explore the clinical application and experience of cardiopulmonary bypass (CPB) diluted autologous transfusion so as to provide clinical rationales for blood conservation.

At Department of Cardiothoracic Surgery, First Affiliated Hospital,Zhengzhou University,the clinical data were reviewed for 5000 CPB surgical patients without banked blood from September 1996 to March 2012 to analyze the clinical application of autologous blood transfusion and summarize the experience of depth hemodilution.No significant changes occurred in blood pressure, central venous pressure or heart rate after blood collection.The values of blood gases stayed within the normal perioperative range.The collected blood and all the remaining blood in CPB machine were reinfused to bring the postoperative level of Hb to over 90 g/L.Only 129 cases on a second thoracotomy for hemostasis required banked blood transfusion because of hemogloblin (Hb) under 70 g/L.

Hb, hematocrit (Hct), pH, base excess (BE), lactate (Lac), cerebral oxygen saturation (rSO₂) and arterial oxygen saturation (SaO₂) indicated a statistically significant during CPB compared with pre-anesthetic (P<0.05).The remaining indicators stayed within the normal perioperative range except for Hb and Hct.There were 146 cases of deaths.And the causes were postoperative low cardiac output (n=34), cerebrovascular accident (n=48), malignant arrhythmias (n=28) and lung infection and multiple organ dysfunction (n=36).The case fatality rate was merely 2.92%.All survivors recovered well and no heart, brain, kidney or other complications occurred.

Hct 15%- 19% depth of hemodilution with proper cooling and anesthetic management during cardiopulmonary bypass is both safe and feasible.And neutralization heparinemia in stored blood before CPB by protamine is one of the effective

of blood conservation.

To explore the surgical treatment and prognostic factors of bronchopulmonary carcinoid tumors (BPC).

The clinical data of 65 patients undergoing surgery for BPC from May 1999 to December 2007 were reviewed retrospectively.The predictors of univariate and multivariate analyses included gender,age,smoking history, pathological type and tumor stage.

The procedures included segmentectomy (n=1), lobectomy (n=36), bilobectomy (n=6), sleeve resection (n=4), pneumonectomy (n=11), carinal resection (n=2), bronchoplastic resection (n=4) and exploratory thoracotomy (n=1).The 1-, 3- and 5-year overall survival rates were 86.2%, 73.8% and 64.6% respectively.Univariate analysis showed that gender (P=0.029),age (P=0.003), smoking history (P=0.039), pathological type (P<0.01), tumor stage (P<0.01), postoperative radiochemotherapy (P<0.01), lymph node metastasis (P<0.01) and surgical type (P=0.042) were prognostic factors.And multivariate analysis revealed that pathological type (P=0.019) and lymph node metastasis (P<0.01) were independent prognostic factors.

Surgery remains a first-choice for BPC.The major resection procedure is anatomical lobectomy or pneumonectomy.Both pathological type and lymph node metastasis are independent prognostic factors.

To evaluate the application value of multiple phase subtraction imaging of dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) in common breast tumor.

All radiographic and MRI data of 72 patients with histological proved common breast tumor were analyzed retrospectively.Prior to an injection of gadolinium-diethylenetriaminepentaacetate (GDDTPA), 3D mask scans were performed.Then 9 consecutive phases of imaging were acquired.After the drawing of time-signal intensity curve, he first and second phases were selected to perform subtraction with the mask, rebuild 3D maximum intensity projection and analyze the application value of subtracted and 3D maximum intensity projection (MIP) reformation images.Chi-square test was used for the analyses of benign and malignant breast tumors.

There were 83 lesions in 72 patients.Among 18 cases of fibro adenoma,23 lesions had no tumor vessel.For the detecting ratio of lesions, the first and second phase subtraction imaging had no statistics difference.Only one intraductal papilloma with ductal carcinoma in situ (DCIS) had tumor vessel in 4 intraductal papilloma patients.Among 5 DCIS cases, one of them had small curved tumor vessel.And 51 lesions were found in 45 infiltrating ductal carcinoma (IDC) cases.Tumor vessels could be seen in all IDC cases.And 37 cases had multiple tumor vessels while 8 cases had single tumor vessel.When the first phase was used for subtraction, 36 lesions were found.On the other hand, 51 lesions were found in the second phase subtraction.There were significant differences between them.

No significant differences exist in benign breast tumor on differential phase subtraction images of DCEMRI, but there are significant differences in malignant breast tumor.It is very important to detect the lesions and show the tumor vessels by rebuilding multiple phase subtraction.

To explore the cortical thickness changes in the patients with type 2 diabetes mellitus and the effects of the insuline therapy on the brain using whole brain cortical thickness mapping.

High resolution three-dimensional T1-weighted fast spoiled gradient recalled echo MRIimages were obtained from 11 normal controls and 11 patients with typeⅡdiabetes at baseline level (before insulin therapy) and follow-up level (after insulin therapy for 1 year).The whole brain cortical thickness map was calculated by the closest point methods.The cortical thickness of each brain region was calculated using the automatic region of interest.The paired t test was performed for the mean cortical thickness.

The mean cortical thickness of bilateral hemisphere and whole brain was decreased in the patients with type 2 diabetes mellitus compared with normal controls (P<0.05).After insuline therapy 1 year, the mean cortical thickness of right hemisphere and whole brain at follow-up was significantly increased compared with that at baseline (P<0.05), and the mean cortial thickness of left hemisphere showed no significant difference between baseline and follow-up (P>0.05).Mean cortical thickness over each brain region demonstratd that increased cortical thickness were mainly located in the left medial orbitofrontal gyrus, bilateral pars opercularis, right supramarginal gyrus, right lateral occipital gyrus, left calcarine gyrus, right fusiform gyrus, bilateral inferior temporal gyri, middle temporal gyri, left caudal anterior cingulate, bilateral entorhinal cortex, left parahippocampal cortex, right insular lobe and right rostral anterior cingulate.

Cortical thinning was investigated in the patients with type 2 diabetes mellitus compared with normal controls, and wide cortical thickening was demonstrated after insuline therapy for 1 years.The precise mechanism should be further explored.

To retrospectively explore the correlation between anterior cruciate ligament (ACL) -ruptured knees, stability of ACL-rupture knee and posterior tibial slope (PTS).

From January 2008 to October 2012,150 knees with ACL rupture underwent arthroscopic surgery for ACL reconstruction.A control group was established for subjects undergoing arthroscopic surgery without ACL rupture during the same period.PTS was measured on a digitalized lateral radiograph.Lachman and mechanized pivot shift tests were performed for assessing the stability of knee.

There was significant difference (P=0.007) in PTS angle between the patients with ACL rupture (9.5±2.2 degrees) and the control group (6.6±1.8 degrees).Only among females, increased slope of tibial plateau had effect on the Lachman test.There was a higher positive rate of pivot shift test in patients of increased posterior slope in the ACL rupture group.Conclusion Increased posterior tibial slope (>6.6) appears to contribute to non-contact ACL injuries in females.And the changes of tibial slope have no effect upon the Lachman test.However, large changes in tibial slope affect pivot shift.

To explore the value of dynamic arterial elastance (Eadyn) in the predication of arterial pressure response to volume loading in shock patients.

A total of 32 patients with pulse indicator continuous cardiac output (PICCO) monitoring at our intensive care unit from January 2011 to December 2012 were retrospectively studied.The decision of fluid replacement was based upon the presence of shock (mean arterial pressure (MAP) ≤65 mm Hg, systolic arterial pressure<90 mm Hg or a decrease of 40 mm Hg from baseline) and preserved volume responsiveness condition with a stroke volume variation (SVV) value≥10%.According to the MAP increase after volume loading, they were classified into MAP responders (≥15%) and MAP nonresponders (<15%) respectively.The goal was to investigate the influencing factors of the changes of MAP after volume loading and predict the arterial pressure response to volume loading.

Significantly different between MAP responders and MAP nonresponders, baseline Eadyn was an effective predictor of MAP increase after volume loading.The area under the ROC curve was 0.95 for the prediction of volume loading on MAP for Eadyn at baseline (P<0.01).A baseline Eadyn value>0.85 predicted a MAP increase after volume administration with a sensitivity of 89.5% and a specificity of 92.3%.

Baseline Eadyn may predict accurately arterial pressure response in MAP to volume loading in shock patients.

To explore our experience of anesthetic management for pediatric congenital laryngomalacia operation.

A total of 27 pediatric patients with congenital laryngomalacia were treated at our hospital between December 2010 and November 2012.All patients were anesthetized by intravenous anesthesia of propofol-remifentanil and spontaneous breathing.Oxygen was insufflated at a rate of 4 L/min through an endotracheal tube near glottis.Propofol was set at a constant rate of 100μg·kg^-1·min^-1. The initial dose of remifentanil at 0.05μg·kg^-1·min^-1 was adjusted in 0.05μg·kg^-1·min^-1 increments to titrate a 50% reduction in baseline respiratory rate.Heart rate (HR), mean arterial pressure, pulse oxygen saturation (SpO₂), respiratory rate (RR), operation time, anesthesia time and remifentanil rate were recorded.Adverse events and interventions were also examined.

Comparison with induction of anesthesia, HR and RR changed significantly intraoperatively (P<0.05).MAP, SpO2 were no significantly change during operation (P>0.05).The induction time was 9-12 min and the highest remifentanil rate stood at (0.18±0.03) μg·kg^-1·min^-1. Body movements occurred in 3 (11%) patients and a bolus of propofol was administered.Desaturation below 95% occurred in 2 (7%) patients in which interventions were offered by decreasing the remifentanil infusion rate.No complications such as cough, hypoxemia, laryngospasm or bronchospasm, nausea or vomiting, arrhythmia were observed.Conclusion Key points of anesthetic management for pediatric congenital laryngomalacia include sufficient preoperative evaluation, spontaneous respiration anesthesia technique with total intravenous anesthesia,suitable anesthesia depth and intensive intraoperative monitoring.

To evaluate the modulation of RhoA/Rho kinase (ROCK),a small Rho GTPase, on migration,invasion and proliferation of fibroblast like synoviocytes (FLS) from rheumatoid arthritis (RA) patients.

RAFLS were collected from active RA patients.And 10% fetal bovine serum (FBS) and interleukin-1β (IL-1β) were used as stimuli in migration and proliferation experiments respectively.RhoA activity was measured by pull down assay while ROCK activity by Western blot.FLSmigration and invasion in vitro were measured by the Transwell chamber method.And thiazolyl blue tetrazolium bromide (MTT) test was used to detect cell proliferation.

There were increased activities of RhoA and ROCK in ex vivo FLS from RA versus OA patients and healthy control.The migrated cell number of FBS-induced, C3-treated and Y27632-treated groups was 85±14,51±15 and 42±11 respectively.The Matrigel invading cell number of 3 groups was 64±13, 39±12 and 26±9 respectively.Statistical differences existed in cell number between FBS-induced, C3-treated or Y27632-treated group (P<0.05) in above migration and invasion experiments.Inhibition of RhoA and ROCK activity also suppressed the cytoskeletal reorganization and proliferation of RAFLS.

Increased RhoA/ROCK activity may contribute to abnormal migration, invasion and proliferation of RA FLS.Thus inhibition of ROCK activity may be a new therapeutic target for RA.

To explore the effects of GRP78 suppression on the sensitivity to cisplatin and elucidate the role and mechanism of GRP78 in ovarian cancer cisplatin resistance.

The GRP78 siRNA expression plasmid was constructed to suppress GRP78 expression.Cell viability was detected by methyl thiazolyl tetrazolium (MTT) assay.Endoplasmic reticulum stress-related apoptosis related protein expressions were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.And cell apoptosis was detected by flow cytometry.

The expressions of GRP78, CHOP and cleaved-caspase 4 were induced significantly by cisplatin (6 mg/L) in SKOV3 cells.And the expression of GRP78 was induced significantly by cisplatin in SKOV3/DDP cells.But the expressions of CHOP and cleaved-caspase 4 showed no significant difference.Inhibition of GRP78 expression and cisplatin combined treatment significantly increased the expressions of cleaved-caspase 4 and cleaved-caspase 3 in SKOV3/DDP cells.Inhibition of GRP78 expression reduced the cisplatin-induced up-regulations of p-Akt and p-mTOR and induced XBP1 mRNA shear expression and CHOP mRNA expression.

Inhibition of GRP78 expression reverses cisplatin resistance in SKOV3/DDP cells.The mechanism may be through the activity of Akt/mTOR signaling pathway, CHOP expression levels and caspase activity.

To explore whether a tissue-engineered construct composed of autogenous endothelial cells, osteoblasts and a new bioresorbable nano-hydroxyapatite/recombinant human-like collagen/polylactic acid (nHA/RHLC/PLA) would enhance bone regeneration and repair femoral head defects in canine models.

The bone marrow stem cells (BMSCs) were isolated from bone marrow of canine ilium and cultured in Dulbecco′s modified eagle medium:nutrient mixture F-12 culture media for 1 week and the second-generation BMSCs were further induced by osteogenic medium (1×10^-8 mol/L dexamethasone, 10 mmol/L B-sodium glycerophosphate and 50μg/ml vitamin C) and by endothelial cell grow medium (vascular endothelial growth factor and basic fibroblast growth factor) for 14 days in vitro.Thus BMSCs were induced into ECs and OBs.After the second passage, cells were digested and collected.And cell density was adjusted to 1.0×10⁶/ml.The cells and nHA/RHLC/PLA scaffold were co-cultured for 2-4 hours then nHA/RHLC/PLA scaffold composites prepared.Cavity defects of 8 mm in diameter and 10 mm in height were made in femoral heads.The nHA/RHLC/PLA scaffold composited with ECs and osteoblasts (OBs)(group A) and composited with OBs (group B) were inserted into different defects while cell-free nHA/RHLC/PLA scaffold served as controls (group C).New bone formation and defect repair were evaluated at 3 and 6 months by radiographic examination, histology and bone histomorphometry.

New bone formation was evident as early as 3 months in groups A, B and C.At 6 months, abundant bone tissue within defects was observed in group A.The control animals with cell-free scaffold showed less bone formation at both timepoints.The scaffold of nHA/RHLC/PLA was degraded and absorbed gradually with the formation of new bone tissues.Histology and bone histomorphometry further revealed significantly increased trabecular bones in group A compared with groups B and C at 6 months postimplantation (P<0.01).Conclusion More abundant new bone tissue may be found in the bone defect areas implanted with osteoblast-endotheliocyte composite than osteoblasts composite and scaffold materials only.ECs and osteoblasts derived from BMSC are ideal seed cells for repairing femoral head defects.

To assess the neuroprotective effect of Tempol,an antioxidant acting as a superoxide dismutase mimic, on white matter lesions and cognitive impairment in rats with chronic cerebral hypoperfusion.

Chronic cerebral ischemia was induced by permanent occlusion of bilateral common carotid arteries (2-VO) in male Wistar rats.The animals were divided into sham operation, saline-treated, Tempol (8 mg/kg) and Tempol (30 mg/kg) groups (n=15 each).Performance of Morris water maze task and Western blot for myelin basic protein (MBP), amyloid precursor protein (APP) and 4-hydroxy-2-nonenal (H NE) modified proteins were analyzed at 6 weeks post-hypoperfusion.Results Tempol reduced the escape latency of Morris water maze post-hypoperfusion in comparison with the saline-treated rats (P<0.05).The mean relative optical density of MBP in the white matter was significantly higher in Tempol (8 mg/kg) group (0.82±0.17) and Tempol (30 mg/kg) group (0.91±0.15) than saline-treated group (0.44±0.13, all P<0.01).The mean relative optical density of APP in white matter was significantly lower in Tempol (8 mg/kg) group (0.55±0.13) and Tempol (30 mg/kg) group (0.46±0.15) than saline-treated group (0.96±0.19, all P<0.01).The mean relative optical density of HNE-modified protein in white matter was significantly lower in Tempol (8 mg/kg) group (0.20±0.035) and Tempol (30 mg/kg) group (0.18±0.031) than saline-treated group (0.29±0.039,all P<0.01).

Tempol ameliorates cognitive impairment by preventing white matter lesions induced by chronic cerebral hypoperfusion in rats.And it may protect white matter lesions in hypoperfused rats through reducing the formation of lipid peroxidation.