OBJECTIVETo explore the value of the application of oil red O staining in spinal cord injury (SCI) of rats.

METHODSWith simple randomization, 24 Spargue-Dawley male rats were divided into normal control group including 6, and SCI group including 18. Spinal cord was transected at spinal lever T10 to build SCI model. Six rats of SCI group were sacrificed randomly at 1, 2, 4 weeks after surgery. After the spinal cord tissue sections were made, oil red O staining methods were used to observe the changes at the end of transected spinal cord. Images were analyzed by Image-Pro Plus 6.0 and SPSS 20.0 software.

RESULTSThe oil red O staining of normal control group showed that white matter surrounded by myelin sheath was clear and obviously distinctive from grey matter. Uneven and strengthened staining in oil O was observed in grey matter of SCI group at 1, 2, 4 weeks post-SCI.

CONCLUSIONIt is a good method to label the myelin sheath in spinal cord and distinct white matter from grey matter by oil red O staining. Analysis of the images showed that lipid may become another target for drugs, which needs more researches.

Animals ; Azo Compounds ; Gray Matter ; pathology ; Male ; Nerve Fibers ; physiology ; Nerve Regeneration ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; pathology ; Staining and Labeling

Objective To analyze the clinical characteristics of leucine-rich glioma-inactivated protein 1 antibody-associated encephalitis.Methods Fourteen patients diagnosed as leucine-rich gliomainactivated protein 1 antibody-associated encephalitis in Capital Medical University Xuanwu Hospital from January 2012 to January 2015 were recruited.The clinical manifestation,brain magnetic resonance imaging,cerebrospinal fluid findings and biochemical examination of these patients were analyzed.Results The case series of 14 patients had an average age of (48.93 ± 15.60) years (range 27-67 years) with a male to female ratio of 2.5∶ 1.All patients presented with short-term memory loss.Nine patients experienced dementia.Additionally,among the 14 patients,12 experienced seizures,8 experienced faciobrachial dystonic seizures,10 had psychiatric symptoms,and 8 showed sleep dysfunction.Two patients were transferred into intensive care unit because of deteriorating symptoms and were provided operated mechanical ventilation.Thirteen of 14 patients exhibited abnormalities in their brain magnetic resonance imaging,with lesions in temporal lobe and hippocampus.Six patients had abnormal cerebrospinal fluid findings,8 patients showed hyponatremia (serum Na+ ＜ 135 mmol/L),while 5 patients co-existed with other autoantibodies.Screening for malignant tumors revealed normal findings.During 2 years follow-up,3 patients relapsed.Conclusions Leucine-rich glioma-inactivated protein 1 antibody-associated encephalitis is an autoimmune encephalitis characterized by short-term memory loss,faciobrachial dystonic seizures and hyponatremia.Lesions in brain MRI always involve in temporal lobe and hippocampus.This disease can relapse and is seldom associated with tumor.

Objective To analyze the improvement of clinical symptoms,relapse and neurological functional recovery and the prognostic factors of anti-N-methyl-D-aspatate receptor (NMDAR) encephalitis.Methods Follow-up was conducted for 51 hospitalized patients with anti-NMDAR encephalitis at the Department of Neurology,Xuanwu Hospital,Capital Medical University from June 2012 to April 2015.The neurological functional recovery was evaluated through modified Ranking Scale (mRS),and the prognostic factors were analyzed.Results Among the 51 patients with anti-NMDAR encephalitis,89％ (45/51) were completely recovered or remained mild neurological dysfunction (mRS score ≤ 2).The prognosis of main clinical symptoms was as follows:78％ (35/44) of the mental and behavior disorders were fully recovered,94％ (32/34) of the seizures were controlled and 65％ (21/31) of the cognitive deficiency were completely recovered;25％ (13/51) of the patients relapsed.Comparison of clinical data of initial on-set among complete recovery patients group (mRS score =0),partial recovery patients group (mRS score =1 or 2) and poor prognosis patients group (mRS score ≥ 3) showed that initial clinical manifestation with memory deficiency (17,9,6 cases respectively;x2 =6.664,P=0.036),involuntary movements(19,4,5 cases respectively;x2 =7.976,P =0.019) and central hypoventilation (5,0,2 cases respectively;x2 =6.124,P =0.047) had statistically significant difference.Conclusions The majority of anti-NMDAR patients have favorable prognosis,but some of the patients may remain various degrees of neurological deficiency,including mental and behavior disorders,cognitive deficiency and seizures.Initial clinical manifestation with memory deficiency,involuntary movements and central hypoventilation may indicate a poorer prognosis.

Objective To evaluate the application of modified Abbe flap in repairing moderate defects of the upper lip and time point to divide the pedicle.Methods Classic Abbe flap was modified in its design,pedicle cutting and dividing time,which was used to repair moderate defect of the upper lip in 12 cases.Surgery was divided into two phases:one with modified Abbe flap surgery was performed for the combined nasal deformity repair simultaneously,and then the pedicle was divided 9days after surgery.Results 12 patients underwent modified Abbe flap.No vascular complications were found in these flaps.Upper lip shape was well and satisfactory functional recovery,corresponding improvement in nasal appearance.Conclusions The surgery that the modified Abbe flap with the pedicle is divided 9 days after surgery is very simple.On one hand,it greatly improves the patient's appearance and function of the upper lip,improve the overall shape of midface,on the other hand,dividing pedicle time is significantly shorter than in the past,specially reducing the suffering of patients and duration.It is particularly suitable for unilateral and bilateral cleft lip of the upper lip on secondary moderate deformities and combined nasal deformities.

Objective To explore the dynamic changes of D-dimers during pregnancy and early puerperium (within 3 days postpartum). Methods A retrospective study was performed among 8 367 healthy women who had term singleton delivery in Women′s Hospital, School of Medicine, Zhejiang University from January 2007 to December 2014. D-dimers concentrations during pregnancy and early puerprium of all the cases were collected. Data of 21 065 D-dimers tests were assigned to 5 groups according to the time of sampling, including early pregnancy (≤12 gestation weeks), middle pregnancy (12-28 gestation weeks), late pregnancy (>28 gestation weeks), 1 postpartum (within 48 hours postpartum) and 2 postpartum (48-72 hours postpartum). The D-dimers concentrations in different groups were compared. The effect of delivery mode on D-dimers of early pureperium was analyzed. The correlation between D-dimers and the thromboembolic disease was also explored. In this study, Student′s t-test and Wilcoxon rank sum test were used for statistical analysis. D-dimers concentration≤0.5 mg/L was used as the normal range. Results (1) D-dimers concentrations during pregnancy were higher than the non-pregnant women (P<0.01), but there was no statistical difference between early pregnancy and late pregnancy (P=0.820). D-dimers concentration in the 1 postpartum group was higher than that of early pregnancy group or late pregnancy group (P<0.01). But in the 2 postpartum group, it was lower than early pregnancy, late pregnancy and 1 postpartum groups. (2)D-dimers in cesarean section cases was significantly higher than in vaginal delivery cases in each period of pregnancy and early pueprium.(3)The 95%CI of D-dimers in early pregnancy, late pregnancy, 48 hours after vaginal delivery, 48-72 hours after vaginal delivery, ≤48 hours after cesarean section, 48-72 hours after cesarean section were 0.58-8.28, 0.47-11.52, 1.04-9.59, 0.87-5.22, 1.07-11.58 and 1.00-6.23 mg/L, respectively.(4)In 6 cases with thromboembolic disease, D-dimers was 6.89-19.89 mg/L, with the mean value of 13.66 mg/L. It was significantly higher than normal range. In 3 cases, all after cesarean section, with lower extremity vein thrombosis within 48 hours postpartum, the D-dimers concentrations, 9.77, 8.65 and 6.89 mg/L respectively, were in the 95%CI of the study population after cesarean section. Conclusions D-dimers concentration of 0.5 mg/L is not suitable for venous thromboembolism screening during pregnancy. D-dimers concentration in pregnancy and early puerprium is higher than non-pregnancy. It increases in the very early period postpartum and decreases with time. D-dimers should not be a routine screening test to exclude thromboembolic disease in pregnant women without high risk factors and clinical manifestation of thromboembolic disease.

Objective To investigate the risk factors and clinical manifestations of placental abruption, and to analyze the causes of missed diagnosis and misdiagnosis. Methods A retrospective analysis was conducted in 135584 women who delivered in Women′s Hospital, School of Medicine, Zhejiang University from January 2005 to December 2015. The diagnosis of placental abruption was made in 1212 cases. According to the consistency of prenatal and postnatal diagnosis, they were divided into 3 groups.(1) The diagnosis was consistent prenatally and postnatally in 715 cases(58.99%,715/1212) as the diagnosis group.(2)In 312 cases (25.74%,312/1212), the diagnosis was made after birth as the missed diagnosis group.(3)In 185 cases (15.26%,185/1212), the diagnosis was made prenatally but excluded after birth as the misdiagnosis group. The disease classification was made, and the risk factors, clinical manifestations, lab results, the time of termination and perinatal outcomes were recorded in the 3 groups. The reasons of missed diagnosis and misdiagnosis were analyzed. Results (1) In the 1212 cases, the diagnosis of placental abruption was confirmed in 1027 cases, with the incidence of 0.76%(1027/135584). The rate of missed diagnosis was 30.38%(312/1027), and the rate of misdiagnosis was 0.14%(185/134557). (2) There were significant differences in the degree of placental abruption among the 3 groups (P＜0.05). (3)Significant differences were found among the 3 groups regarding the ratio of hypertensive disorders, trauma, induced labor and advanced maternal age (all P＜0.05). (4) There were statistically significant differences among the 3 groups regarding the incidence of vaginal bleeding, persistent abdominal pain and uterine tenderness, bloody amniotic fluid, increased uterine tension and stillbirth (all P＜0.05). (5) There was no significant difference in the rate of abnormal fetal heart rate mornitoring among the 3 groups (P=0.22). The differences were statistically significant among the 3 groups when regarding the incidence of abnormal ultrasound finding and abnormal blood coagulation (P＜0.01), with the highest incidence of abnormal ultrasound in the diagnosis group (68.1%) and the highest incidence of abnormal coagulation in the misdiagnosis group (24.9%). (6)There was statistically significant difference among the 3 groups when comparing the ratio of termination of pregnancy within 24 hours (P=0.01). (7) There were statistically significant differences among the 3 groups when the ratios of postpartum hemorrhage, DIC, neonatal asphyxia and perinatal death were compared (all P＜0.05). The highest incidence of postpartum hemorrhage was in the diagnosis group (17.9%) and the lowest was in the misdiagnosis group (5.4%). The highest incidence of DIC was in the diagnosis group (3.9%) and the lowest was in the misdiagnosis group (0). The highest incidence of neonatal asphyxia was in the diagnosis group (30.6%) and the lowest was in the misdiagnosis group (7.6%). And for perinatal death, the highest incidence was in the diagnosis group (12.6%), the lowest was in the misdiagnosis group (2.2%). Conclusions Placental abruption could be misdiagnosed when depending on risk factors, such as trauma. And it could be missed diagnosis during the induction of labor. Uterine contraction, abnormal fetal heart rate mornitoring, abnormal ultrasound and abnormal coagulation function are important in the diagnosis of placental abruption.

Objective To search for a good method for generation of hematopoietic stem cells (HSC) by embryonic stem cells (ESC),and to investigate the potential of HSC derived from ESC to reconstruct hematopoiesis in vivo.Methods Using a three-step method to induce a mice ESC line, E14.1-into HSC.And identifying HSC by flow cytometry analysis cell markers with CD34 +/Sca-1+, then HSC (1×109L-1)from ESC differentiation were injected into severe combined immunodefieency (SCID) mice for observing teratoma formation.To validate function of HSC by colonngenic cells assay and to reconstitute the hematopoiesis in lethally irradiated mice.Results Combining to use more hematopoietic stimulating factor to availably promote the El4.1 cell into embryoid body (EBs) with abundant hematopoietic progenitors.EBs were induced after 14 day to fast differentiate for HSC with peak percentages of CD34+/Sca-1+ cells reached to (13.72±2.07)%.To harvested ceils from EBs by day 14 for second -step HSC differentiation, percent of CD34+/Sca-1+ cells rise to(24.62±2.50) % after day 16 induction.Cloning forming units (CFU) analysis showed that more Erythro -myeloid cloning generation was observed at this period.Cells obtained in the second step are subsequently plated on monolayer of mice bone marrow stromal cells, in the presence of TPO, FLt3 ligand and superoatant of mice fetal liver stromal cells, cultured for additional 15 days, followed fast expansion of CD34+/Sca-1+ to maximally (58.64±4.20 ) % with more CFU-E, CFU-GM and CFU-GEMM population.Wright-Giemsa stain showed that its had the character of hematopoietic progenitors.Cells from the third-step were injected into SCID mice, but no teratomas were recovered in 2 mices after 6 weeks.Positive selection of CD34+/Sca-1+ cells by magnetic sorting from the third - step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 days after transplantation, with 71.4% successful engraftment rate.And 3 recipients showed that the cell population of the peripheral blood leukocytes,red cells, hemoglobin approached normal index at 40 days after transplant, hut followed relative'slow renew in platelet count.Survival rate of transplant group is 43.0%, compared to 100% mortality in control mice.Karyotyping assays confirmed female mice with XY.Conclusions The results showed that the three-step differentiation and the culture conditions described here good support differentiation of ESC into HSC.HSC derived from ESC were safe without teratomas formation in body, and its can reconstruct hematopoiesis.

Objective To investigate and analyze the behavior and therapeutic status of rheumatoid arthritis (RA) patients. Methods Out patients diagnosed with rheumatoid arthritis in our hospital from May to August 2007 were enrolled. The data including sex, age of onset, site of first hospitalization and medication status were collected and analyzed. Results In this 181 RA patients, the mean age of onset was (53±11) year-old, mean history duration was (10±8) years, the ratio of male to female was 1:4.2. The orthopedics department was the most common site of first hospital visit (32.0%, 58/181) and rheumatology department was the most common site to clarify the diagnosis (62.4% ,113/181). The diagnosis delay caused by patients themselves was (5.9±17.2) months and the delay caused by doctors was (9.0±22.0) months. More than half of the patients were not treated appropriately before they came to our hospital. Methotrexate was the most commonly used DMARDs (67.3%), followed by leflunomide (46.4%), sulfasalazine (37.5%) and hydroxyehloroquine (19.6%). Conclusion In this cohort, the proportion of patients who come to rheumatology department immediately after disease onset is low. There is delay between symptoms and final diagnosis. More than half of the patients are not treated appropriately.

Objective To investigate the practical anatomy and clinical applications of the lateral sural nerve-lesser saphenous vein vascular plexus island flap with median sural nerve preserved.Methods Ten lower limbs injected with color red latex and 20 fresh limbs were observed.Based on anatomical studies,the island flap pedicled with the vascular axis of lateral sural nerve and lesser saphenous vein was designed for clinical application in 63 cases.In procedure,the median sural nerve were preserved in donor site and the distal end of the lesser saphenous vein were not ligated.Results The accompanying arteries formed a favourable vascular plexus around the nerve and the vein,lied along the lateral sural nerve and the lesser saphenous vein and nourished the skin through cutaneous branches.The distance of the lateral ankle to the confluent of lateral and median sural nerve was (8.5± 0.8) cm.63 patients with soft tissue defects in the distal leg and foot were repaired.Their appearance and function were satisfaction without feeling lose in the heel and lateral side of the foot.Conclusions The lateral sural nerve-lesser saphenous vein vascular island flap is a reliable source of soft tissue to cover defects in the lower leg and foot.This flap has sufficient blood supply and high survival rate without the sacrifice of the median sural nerve.

Objective and Methods: Thirty-three patients with peri-menopausal syndrome were treated by electroacupuncture of points Guanyuan (CV 4) and Sanyinjiao (SP 6) according to the principle of regulating and tonifying the Thoroughfare and Conception vessels, and the curative effects were analyzed.Results and Conclusion: The results showed that there was a significant difference in the clinical symptoms of the patients with peri-menopausal syndrome between pre- and post-electroacupuncture of points Guanyuan (CV 4) and Sanyinjiao (CV 6) and the clinical effective rate reached 93.9%. The measurement of sexual hormones showed E2, FSH and LH levels were better after treatment than before treatment in the patients with peri-menopausal syndrome. The analyses indicated that electroacupuncture of points Guanyuan (CV 4) and Sanyinjiao (CV 6) could relieve the clinical symptoms in the patients with peri-menopausal syndrome and improve the patient's life quality.