Plastrous of Chinemys reevesii (Gray) and Morenia ocellata (Dum.et Bid) are compared and identified by characte rization, TLC and UV spectroscopy.The method is simple and reliable.

BACKGROUND:Currently, bone graft is mainly used for repair of bone defects, and tricalcium phosphate is the most used artificial bone material. But the effectiveness of the tricalcium phosphate bone graft is stil controversial, and there is also no detailed report about its function during the healing of bone defect. ＜br> OBJECTIVE:To observe the concentration changes of bone morphogenetic protein-2 and vascular endothelial growth factor as wel as bone healing after tricalcium phosphate graft in bone defects. ＜br> METHODS:Forty-eight C57 mice were randomly divided to experimental group and control group. A 2-mm-long diaphyseal segment and periosteum from the middle of the right femur was cut to prepare unilateral bone defect models. Tricalcium phosphate bone graft was used in the experimental group, and no bone graft in the control group. During the fol owing 4 weeks, X-ray examination was done once a week to observe the bone healing, and then the animals were executed for col ecting samples in the graft area. The concentrations of bone morphogenetic protein-2 and vascular endothelial growth factor in samples which were taken from the bone graft area were determined by using ELISA assay. ＜br> RESULTS AND CONCLUSION:X-ray films showed that 2 weeks later, bone fracture healed mostly in the experimental group except a smal part of cortical bone;3 weeks later, bone fracture was basical y healed, and only a smal amount of tricalcium phosphate remained;4 weeks later, bone fracture was completely healed, and the cal us grew obviously, and the tricalcium phosphate was nearly absorbed. In the control group, the fracture line was stil visible at 1-2 weeks, but it became vague at 3 weeks;then, the fracture was healed at 4 weeks except some of the cortical bone. The levels of bone morphogenetic protein-2 and vascular endothelial growth factor were significantly higher in the experimental group than in the control group at different time points (P＜0.05). These results suggest that tricalcium phosphate bone graft can up-regulate the expression of bone morphogenetic protein-2 and vascular endothelial growth factor and accelerate bone healing.

Objective To investigate the electromyngraphy eharacteristies of Parkinsen's disease (PD) by studying tremor frequency, intensity and drug response. Methods Sixty-six patients with PD hospitalized during March 2005 to May 2006 were re-evaluated and recruited into the study. A ten-channel electromyography recorded the frequency and intensity of the rest tremor and postural tremor. Tremor analysis soft was used to statistically analyze differences between the two types of tremor, correlation between the frequency and intensity, and changes after regular administration of dopaminergic medication. Results The frequencies of rest tremor ((4. 79±0.69 ) Hz) and postural tremor ( (5.54±1.18) Hz ) showed statistical differences (F = 33.5, P<0. 01), but the intensity of rest tremor (0.016-421.700 (rag)2) and postural tremor (0.062-217.900 (mg)2) didn't show statistical differences (F=1.917, P>0.05 ). No regression relation was revealed between the frequency and intensity of tremor (r=0.09, P>0.05). After dopaminergic medication treatment, the intensity declined significantly from 0.016-421.700 (mg)2 to 0.027-82.200 (mg)2(F=5.179,P <0.05), but the difference of frequencies of tremor between before ( (5.09±0.85) Hz) and after treatment ( (5.20 ± 1.08 )Hz) was not significant (F = 0.474, P > 0.05). Conclusions There may be different mechanisms for the rest tremor and postural tremor in PD. There is no correlation identified between the frequency and intensity in PD and this is may be resulted from variability of tremor intensity. Dopaminergic can relieve the symptom of tremor, but it doesn't reduce the frequency of tremor.

BACKGROUND:Systemic erythematosus lupus (SLE) is closely associated with activity of dendritic cells (DCs) function.DCs derive from hematopoietic precursor cells (HPCs) in vivo and many factors might affect the differentiation and maturation of DCs.The cytokine network was in disturbance and the levels of various cytokines were anomalous in SLE serum.Previous studies mainly addressed effects of single factors.OBJECTIVE:To investigate the effects of joint action of serum interferon (IFN)-α and interieukin (IL)-6 in the serum of SLE patients on the differentiation and maturation of DCs derived from CD34~+ HPCs.DESIGN,TIME AND SETTING:The grouping,controlled and completely random designed study was performed at the Department of Microbiology and Immunology,Nanjing Medical University from May 2006 to October 2008.MATERIALS:Peripheral blood samples were collected from 50 SLE patients from the Department of Rheumatology,First Affiliated Hospital of Nanjing Medical University and 30 healthy volunteers from Nanjing Medical University.Cord blood samples were collected from 15 full-term normal delivery neonates from Nanjing Maternity and Children Health Care Hospital Affiliated to Nanjing Medical University.Informed consent was obtained from all patients and their family members.METHODS:Peripheral blood samples were collected from SLE patients and healthy volunteers.According to serum IFN-α and IL-6 concentrations of normal person,95% reference value range of serum IFN-α and IL-6 concentrations were calculated by the method of percentiles.Above this range represented abnormal increase.Cord blood mononuclear cells were harvested by Ficoll-Hypaque density gradient centrifugation.CD34~+HPCs were purified from cord blood by magnetic cell sorting system (MACS),and cultured to differentiate to DCs.There were 6 groups in this study.Normal serum,SLE serum with elevated levels of IFN-α,SLE serum with elevated levels of IL-6,SLE serum with elevated levels of IFN-α and IL-6,SLE serum with anti-IFN-α and anti-IL-6 neutralizing antibodies,or normal serum with exogenous IFN-α and IL-6 were respectively added to the culture medium in each group.10% volume fraction serum was used in each group,for totally 14 days incubation.MAIN OUTCOME MEASURES:The phenotype of DCs was analyzed by flow cytometry (FCM).Cytokine production was assessed by ELISA.The capacity of DCs to stimulate allogenic T lymphocyte proliferation and to regulate T cell differentiation was evaluated in a mixed lymphocyte reaction (MLR).RESULTS:Compared with normal serum,HLA-DR,CD80 and CD86 expression was significantly increased in the medium containing SLE serum with elevated levels of IFN-α,SLE serum with elevated levels of IL-6,SLE serum with elevated levels of IFN-α and IL-6 and normal serum with exogenous IFN-α and IL-6 (P＜0.05),whereas IL-12 levels were significantly decreased (P＜0.05),and capacity of DCs to stimulate allogenic T lymphocyte proliferation was significantly enhanced (P＜0.05).Above-mentioned indexes recovered to a normal level in the medium containing SLE serum with anti-IFN-α and anti-IL-6 neutralizing antibodies.Compared with normal serum,proportion of both CD3~+CD8~-IFN-γ~+ and CD3~+CD8~+IFN-γ~+ T cell subsets was significantly reduced in the medium containing SLE serum with elevated levels of IL-6 (P＜0.05).However,proportion of both CD3~+CD8~-IFN-γ~+ and CD3~+CD8~+IFN-γ~+ T cell subsets was significantly increased in the medium containing SLE serum with elevated levels Of IFN-α,SLE serum with elevated levels of IFN-α and IL-6,normal serum with exogenous IFN-α and IL-6 (P＜0.05).CONCLUSION:The joint action of IFN-α and IL-6 in SLE serum promotes the differentiation and maturation of DCs derived from CD34+HPCs and affects the DC-mediated T cell differentiation,which might contribute to the pathogenesis of SLE.

[Objective] To investigate the effect of mitogen-activated protein kinases (MAPK) on cerebral ischemia induced by photothrombosis in Swedish amyloid precursor protein (APP/SWE) transgenic mice.[Methods] In APP/SWE transgenic mice and non-transgenic mice (n ＝ 12,respectively),photothrombotic stroke was induced,on 7 d after cerebral ischemia,the amount of the survival neuron in the penumbra was counted using Nissl staining (n ＝ 6),and the activities of p38MAPK and JNK were measured by Western blot (n ＝ 6).[Results] On 7 d after cerebral ischemia,ratio of amount of survival neuron over the penumbra in hippocampus in the ischemic side to that in the non-ischemic side in the non-transgenic mice group (78.3 ± 1.3)% was significantly higher than that in the APP/SWE transgenic mice group (70.5 ± 1.4)% (P ＜ 0.05);compared with the non-ischemic hemisphere,the activities of p38 MAPK and JNK increased significantly in the ischemic hemisphere in the APP/SWE transgenic mice group (P ＜ 0.05),whereas,there was no significant difference between ischemic and non-ischemic hemisphere in the non-transgenic mice group (P ＞ 0.05).[Conclusion] Photothrombosis causes more severe damage in the APP/SWE transgenic mice group than that in the non-transgenic mice group.The possible mechanism includes the increased activities of MAPK which enhance the process of neuronal cell apoptosis.

Objective:To analyze risk factors of breast ductal carcinoma in situ (DCIS) with microinvasion (DCIS-MI) and explore suitable axillary lymph node surgery treatment for patients with DCIS-MI. Methods:The clinical characteristics, such as age, menopausal status at diagnosis, size of breast mass, and pathology reports of 45 patients with breast DCIS or DCIS-MI treated at Jinling Hospital, Medical School of Nanjing University from February 2013 to February 2016, were retrospectively collected and analyzed statistically to deter-mine the risk factors associated with microinvasion. Results:Premenopause (P=0.006), tumor size≥3.15 cm (P=0.006), and family his-tory of malignant tumor (P=0.002) were proven risk factors of DCIS-MI. Conclusion:Patients with clinical palpable axillary mass, pre-menopause, large breast mass, and family history of malignant tumor demonstrated high possibility of DCIS-MI. Hence, sentinel lymph node biopsy should be performed. Axillary lymph node dissection is highly recommended to patients whose main symptom is palpable axillary mass.

BACKGROUND: Blood stem cells (BSCs) are the most primitive cells in the immune system and can be differentiated into many kinds of cells. As the regulatory cells of immune response, dendritic cells (DCs) have been attracted more and more attention in the field of autoimmune diseases. Due to different resources of precursor cells, DCs have different cytokines, ideal cytokine matching,applying orders, and experimental cultured conditions. Furthermore, development, phenotypic expression, and mature degree are still different.OBJECTIVE: To investigate the influence of tumor necrosis factor- α (TNF- a ) and interleukin-4 (IL-4) on the culture system and provide improved method for inducing functional DCs in vitro, derived from cord blood CD34+ hematopoietic precursor cells (HPC).DESIGN, TIME AND SETTING: Observational study, which was performed in the Department of Microbiology and Immunology, Nanjing Medical University from March 2005 to November 2005.MATERIALS: The cord blood was collected from neonatal umbilical cord in the 81 Hospital of Nanjing City. CD34 monoclonal antibody coated magnetic bead system (MACS) was provided by Miltenyi Biotec Company, Germany; human recombinant granulocyte-macrophage colony-stimulating factor (rhGM-CSF), human recombinant interleukin-4 (rhlL-4), and human recombinant tumor necrosis factor-α (rhTNF-α) by Pepro Tech Company, USA.METHODS: CD34+HPC were isolated and purified from umbilical cord blood by using a high-gradient magnetic cell sorting system (MACS). Then the cells were cultured with different culture medium which contained different combinations of cytokines:GM-CSF and TNF-α (GT) or GM-CSF, TNF-α and IL-α (GTI) in order to be induced to differentiate into dendritic cells (DCs).The DCs derived from CD34+HPC were identified for their morphology and phcnotype by FACS and laser scanning con_focal fluorescence microscopy, and also for their abilities of inducing proliferation of allogenic T cells by 3H-TdR incorporation assay.RESULTS: The purity of selected CD34+ cells with MACS was more than 90%. DCs could be obtained from CD34+HPC by the culture in presence of GM-CSF and TNF-α or GM-CSF, TNF- o and IL-4. With the time of culture lasting, the cells expressed lower level surface antigen of CD34 and HLA-DR (P ＜ 0.05), and possessed the phenotypes of DCs characterized by higher expression of CD80, CD86, CD83 and CDIa. At 13-15 days, the cells possessed higher level of phenotypes of DCs compared to 7-9 days and 10-12 days. DCs induced with GTI culture system expressed higher levels of surface antigen CD80, CD86, CD83,CD 1 a and a lower level of CD 14 than those induced with GT culture system. DCs displayed typical morphology and property and expressed higher level surface antigen of CD86 and CD80, especially expressed CD86 when they were induced with proper cytokines of GM-CSF and TNF- α added at 0 hour, and of IL-4 added at 48 hours.CONCLUSION: DCs can be generated from CD34+HPC by proper culture system, the design of GM-CSF + TNF- α + IL-4(GM-CSF and TNF- α were added at 0 hour, IL-4 was added at 48 hours) is preferred.

[ ABSTRACT] AIM:To explore the role of superoxide dismutase ( SOD) and malondialdehyde ( MDA) in chronic pancreatitis ( CP) induced by dibutyltin dichloride ( DBTC) combined with ethanol, and the mechanisms for prevention and treatment of pancreatic fibrosis by Chaihushugansan.METHODS: The KM mice were randomly divided into control group, CP group ( DBTC combined with ethanol) and Chaihushugansan group ( CP+Chaihushugansan) .Except for control group, the mice in other groups were intravenously injected in tail with DBTC (8 mg/kg) and drank 10% ethanol.The mice in Chaihushugansan group were administered intragastrically with Chaihushugansan (6 g· kg-1 · d-1 ) at the follow-ing experimenal period.Before modeling and 1 week, 2 weeks, 4 weeks and 8 weeks after modeling, the mice were anes-thetized and sacrificed.The activity of amylase and the content of hyaluronic acid in the serum were measured.The mor-phology and the degree of fibrosis in the pancreas were observed by HE staining.The activity of SOD and the level of MDA in the pancreas homogenate were analyzed.The protein of pancreas was extracted to detect the expression of type I collagen by Western blotting.RESULTS:DBTC combined with ethanol induced CP with increased serum amylase and hyaluronic acid levels, while the serum amylase and hyaluronic acid levels in Chaihushugansan group were significantly lowered ( P<0.05).In 1 week, 2 weeks, 4 weeks and 8 weeks, the pancreas were obviously injured and appeared different degrees of fibrosis.The content of MDA and the expression of type I collagen in the increased significantly, but the SOD was de-creased.In Chaihushugansan group, the pathological damage and the degree of fibrosis of the pancreas were improved.The level of MDA and type I collagen expression in the pancreas were significantly reduced, but the SOD was increased.CON-CLUSION:The oxidative stress may take part in the development of CP.Inhibition of oxidative stress in the pancreas is one of the mechanisms that Chaihushugansan attenuates the development of CP.

Objective:To compare the survival and prognostic factors of intraoperative radiotherapy (IORT) and postoperative radiotherapy (PORT) in female patients, aged≥50 years, diagnosed with node-negative breast cancer (≤ 3 cm in size).Methods:Clinical data of eligible early breast cancer patients between 2010 and 2015 were obtained from the SEER database. Patients were divided into the IORT and PORT groups according to the radiotherapy record and propensity score matching (PSM) was subsequently conducted. Kaplan-Meier curve was used to evaluate the overall survival (OS) and breast cancer-specific survival (BCSS) between two groups and Cox proportional hazard regression analysis was used to explore the risk factors of clinical prognosis.Results:7 068 patients were included after PSM. The median follow-up time was 32.0 months. The 5-year OS rates in the IORT and PORT groups were 96.8% and 93.8%, respectively. Univariate Cox analysis showed that radiotherapy, age, histological grade, T stage, estrogen receptor (ER) status and progesterone receptor (PR) status were the independent risk factors for OS, and histological grade, T stage, ER status, PR status and chemotherapy were the independent risk factors for BCSS. Multivariate Cox regression analysis demonstrated that patients who received IORT had better OS than PORT counterparts ( P=0.020). Besides, patients aged≥60 years obtained worse OS than those aged<60 years ( P=0.003). Patients with T 2 stage or ER-negative tumors had worse OS than those with T 1 stage tumors ( P<0.001) or ER-positive tumors ( P=0.001). Patients with grade Ⅲ-Ⅳ tumors achieved worse BCSS ( P=0.004). Subgroup analysis showed that IORT yielded better OS for elderly patients (≥60 years), grade Ⅲ-Ⅳ tumors, infiltrating duct carcinoma, T 2 stage tumors, ER-positive tumors, PR-positive tumors and patients without chemotherapy. Conclusions:IORT may bring benefit for highly selected patients with low risk of recurrence, which is not inferior to PORT in terms of short-term survival. Prospective studies with longer follow-up time are needed to confirm the findings.

Objective To compare the clinical efficacy of high-flow nasal cannula oxygen therapy (HFNC) with non-invasive positive pressure ventilation (NPPV) in patients with traumatic cervical spinal cord injury complicated with acute respiratory failure (ARF).Methods A prospective randomized controlled trial was performed in EICU of the First Affiliated Hospital of Zhengzhou University from May 2016 to January 2018.One hundred sixty-eight consecutive patients with traumatic cervical spinal cord injury complicated with ARF,who did not respond to conventional oxygen therapy,were assigned to the HFNC or NPPV treatment group sequenced by the random number table.The baseline clinical characteristics of randomized participants and respiratory frequency (RR),PaCO2,mean arterial pressure (MAP) at 1,12,24,48 h after treatment were evaluated.Comfortable scale,tracheal intubation rate within 28 d,duration of mechanical ventilation,length of stay in ICU and mortality rate were compared as well.Results There was no significant differences in baseline clinical characteristics,such as sex,age.between the two groups (P＞0.05).RR and PaCO2 were lower in the HFNC group at all time point.In addition,the HFNC group had significantly lower PaCO2 than the NPPV group at 24 and 48 h after treatment (P＜0.01);Oxygenation index (PaO2/FiO2) was improved in both groups,and the HFNC group had superior oxygenation index than the NPPV group at 12,24,48 h after treatment (P＜0.01).Furthermore,the HFNC group had better comfort scale (6.93±0.71 vs 4.29±0.93,P＜0.01),shorter length of stay in ICU and duration of mechanical ventilation compared to the NPPV group (P＜0.01).There was no significant differences in tracheal intubation rate and mortality rate between the two groups (P＞0.05).Conclusions In addition to the superior efficacy in improving respiratory function and shortening length of stay in ICU,HFNC was well tolerated by patients with traumatic cervical spinal cord injury complicated with ARF,and could be recommended in clinical practice.