AIM: To investigate the process of human bone marrow stromal cells (hBMSCs) differentiation into neural-like cells and to determine the role of 26S proteasome in neuronal differentiation. METHODS: Purified hBMSCs were treated with β-mercaptoethanol (β-ME) for 1 day and retinoic acid (RA) for 3 days, followed by growth factor (10 μg/L bFGF or 20 μg/L NGF) for another 3 days. Immunofluorescence was performed to detect the expression of nestin (a neural precursor cells marker), Tuj1 (a premature neuronal marker), and neurofilament (NF, a mature neuronal marker) at all stages of induced differentiation. Immunostaining and RT-PCR were used to analyze the expression of 26S proteasome during neuronal differentiation of hBMSCs. To further confirm the role of 26S proteasome in hBMSCs differentiation, cells were treated with β-ME/RA and then followed by protesome inhibitor MG132 and growth factor. Immunostaining was performed to detect NF-positive cells. RESULTS: Quantification results showed that the untreated cells were almost never positive for nestin, Tuj1 and NF. After treated with β-ME/RA, the numbers of nestin-positive cells (34.41%±1.27%) and Tuj1-positive cells (27.79%±1.27%) were increased. Notably, the numbers of NF-positive cells were significantly increased to 56.72%±2.4% after induction with β-ME/RA/GF. Immunofluorescence analysis showed that undifferentiated hBMSCs cells were weakly stained by antibody against 26S proteasome, but the numbers of cells with high-intensity of 26S proteasome were increased after treated with β-ME/RA. The RT-PCR result of 26S proteasome further confirmed that the mRNA level of the cells differentiated by β-ME/RA (1.33), as well as by β-ME/RA/GF (1.77), was significantly increased compared to the undifferentiated cells. Moreover, hBMSCs incubated with protesome inhibitor MG132 significantly decreased the numbers of NF-positive cells (37.59%±1.52%). CONCLUSION: After induction with β-ME/RA/GF, hBMSCs can be differentiated into neural-like cells, which is concomitant with the increase in 26S proteasome expression. Inhibitor of 26S protesome prevents hBMSCs differentiation, suggesting that 26S proteasome may be involved in the differentiation of hBMSCs into neural-like cells.

BACKGROUND: Caspase family is viewed as the executive factor of cell apoptosis. Neuronal apoptosis happens probably after spinal cord injury.OBJECTIVE: To observe the changes in caspase-3 expression after spinal cord injury in rats so as to probe into the relationship between it and neuronal apoptosis and provide the evidence on the prop e r time window of intervention on alleviating secondary spinal cord injury.DESIGN: Self-control and mutual-control were designed in animal experiment.SETTING: Department of Traumatic Surgery and Department of Orthopedics of Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Experiment Room of Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology from January to December 2001, in which, 54SD rats were employed, of either sex, mass weighted varied from 220 to 250 g and provided from Animal Experimental Center of Tongji Medical College of Huazhong University of Science and Technology.rats were divided into the control and injury group. Laminectomy was only done on Ts and T9 in the control and the injury group was subdivided into 9 subgroups, in which, the materials were collected on the 4th and 8th hours and on the 1st, 2rd, 3rd, 7th 14th and 21st days successively, 6 rats in each one. After abdominal anesthesia with 30 g/L pentobarbitol sodium,sternal cord on T8 andT9 segments were exposed with Nystrom method and 50 g weight compressed the front middle region of the spinal cord of such segments with arch smooth metal pad 2.2 mn×5.0 mm for 5 minutes. After injury, artificial bladder urination was done 3 times at 10:00, 16:00and 22:00 successively everyday till the bladder reflex was established.cord was collected at various time spots after spinal cord injury. 4 pieces of spinal cord tissue masses from each group, about 8mm in length, were embedded with paraffin and sectioned continuously. Afterwards, HE staining, immunohistocheistry and TUNEL (TdT-mediated dUTP-biotin nick end labeling) were performed successively. Two rats were sacrificed on ice in each group and central tissue of injured spinal cord was placed in expression was assayed with immunohistochemistry method, neuronal apoptosis was assayed with TUNEL method and linear correlation was used to analyze the correlativity between caspase-3 expression and neuronal apoptosis.pase-3 expression after spinal cord injury in rats of each group.RESULTS: Six rats were maintained in each group and included in result optic microscope: Extensive hemorrhage appeared in 1 hour in injured segment. In 4 to 8 hours, spinal structure began destructive and a large amount of neuronal death appeared. In 24 hours, the destruction of spinal cord became severe and in 7 to 21days, the range of injury was defined with immunohistochemistry in rats of each group: Very few caspase-3 expressions (2.1±0.5) presented in neurons of spinal cord in normal rat. In 8hoursafter spinal cord injury, caspase-3 expression of positive neurons was increased remarkably (89.2±10.5) and up to the peak (189.6±12.7) in 3 days. Caspase-3 expression of positive cell and apoptotic cell appeared alexpression assayed with transcription-polymerase chain reaction (RT-PCR)in rats of each group: Caspese-3 mRNA (0.442±0.024) began increased in 4h, was up to the peak (0.634±0.028) in 48 hours and was restored to be normal (0.351±0.013) in 7 days, which appeared early than apoptosis, indicating positive correlation with the level of neuronal apoptosis (r=0.622).In the control and 4 hours group, stained cell was seen occasionally and positive cell appeared 8 hours later, mainly localized in gray matter. Afterwards, positive cell was increased and up to the peak in 3 days. In 7 days,positive cell of apoptosis and staining was decreased gradually in gray matter, mainly around the white matter. Little amount positive cells appeared on the 14th day and 21st day.CONCLUSION: In normal spinal cord tissue, caspase-3 existed in form of zymogen with very low activity. Caspase-3 is enhanced in expression after spinal cord injury in rats, expresses in large amount in 8 hours and is up to the peak in 24 to 48 hours, which is overlapped in time with positive apoptotic cell assayed with TUNEL and concerning to the localization, it is in conformity with positive apoptotic cell of spinal cord injury compared with positive cell of caspase-3. It is indicated that caspase-3 is involved in regulation of cell apoptosis after spinal cord injury. It is seen in this experiment that the time from spinal cord injury to the activation of caspase-3 is the time window of treatment for cell apoptosis intervened by spinal cord and alleviating secondary spinal cord injury. It is suggested that genetic intervention or specific caspase-3 inhibitor should be applied in 48 hours.

This study was based on the project prophase of expert consultation and literature consult, and aimed to analyze the medicine establishment from the perspective of traditional Chinese medicine (TCM) doctors, in order to further study the influence of medical insurance on the development of TCM. Detailed suggestions were made in or-der to promote the benign development trend of TCM and medical insurance. Questionnaire survey was used in the investigation on perceptions of TCM related with medical insurance among 253 TCM doctors in appointed medical institution of medical insurance. Data was processed with frequency statistics. The results showed that after becoming the appointed medical institution of medical insurance, the medical insurance patients have become the main service group (73.5%) of hospitals (68.4%) and incomes of TCM doctors (41.9%) have increased, which contribute to TCM hospitals of becoming bigger and stronger (63.6%). The medical insurance patients have chosen TCM treatment main-ly for its curative effect (37.9%) and safety (24.1%). The influence of reimbursement ratio in medical insurance was relatively low (1.2%). The reimbursement range of TCM medical insurance needs to be further expanded, such as Chinese medicine nosocomial preparation (90.5%), decoction preparation fee (78.3%), and etc. It was concluded that under the background of universal health coverage, medical insurance plays a more and more important role in the development of TCM. And TCM should ensure its safety and improve its curative effect through the standardization of TCM and other measures. Meanwhile, the formulation of medical security policy and medical insurance management service standards should also consider the characteristics of TCM, and encourage the service of TCM.

Alzheimer’s disease (AD) has become one of the worldwide critical diseases which seriously threaten the health of the elderly. Exploring and developing medicine with high efficiency and low toxicity for AD patients is one of the vital medicine issues. Traditional Chinese medicine has high research and development value in the prevention and treatment of AD. Currently, increasing researches have proved that Ginseng Radix et Rhizoma, as a kind of traditional and valuable Chinese herbal medicine, shows effects on improving learning and memorizing ability and prevention and treatment of AD. This article reviewed Ginseng Radix et Rhizoma extract improving symptoms of AD and its mechanism of action in detail, with a purpose to provide references for Ginseng Radix et Rhizoma extract improving learning and memorizing ability and prevention and treatment of AD in clinic.

Objective To investigate the role of a small RNA interference against VEGF in radiation sensitivity in melanoma .Methods A375 human melanoma cell lines were transplanted into nude mice ,which with malignant melanoma were randomly divided into control group , VEGF negative plasmid group and VEGF positive plasmid group, followed by 4Gy irradiation twice a week for 2 weeks.The volume of tumor was calculated twice a week, the area of tumor necrosis was assayed by HE,the expression of VEGF in tumor was determined by Western-blot and Immunohistochemical. ResuIts The expression of VEGF in VEGF positive plasmid group decreased significantly (P<0.05), VEGF positive group had more tissue necrosis, tumor growth was significantly inhibited (P<0.05).ConcIusion siRNA-VEGF in tumor injection liposome encapsulated in malignant melanoma has a role in the radiation sensitization, which provides an experimental basis for the clinical development of targeted therapy combined with radiotherapy for the treatment of VEGF gene.

The development, application and revision of the clinical practice guideline (CPG) in traditional Chinese medicine (TCM) are a whole thing. However, the development and revision of TCM CPGs have been influenced due to lack of TCM CPGs reporting and feedback channel. Therefore, during the TCM standardization network establish-ment, we studied the application model of TCM CPGs with spontaneous reporting network, in order to provide the ba-sis for further TCM CPGs development and revision.

Objective To observe the best compatibility proportion and action mode of protective effects of combined baicalin and taurine in Alzheimer’s disease (AD) model.Methods Aβ1-42 was injected through lateral ventricles to establish AD rat models. SD rats were randomly divided into sham-operation group, model group, aricept group, combined baicalin and taurine in different proportions group. Administration groups were treated with different medicines, while sham-operation group and model group were treated with the same amount of normal saline for 40 days. The step-through latency and the times of mistakes were detected through step-through test;the escape latent and the times of crossing target quadrant were detected through Morris water maze. Furthermore, the human neuroblastoma SH-SY5Y cells were induced by 5 μmol/L Aβ1-42 for establishing a cellular AD model. The AD cellular model was treated with baicalin and taurine compatibility in different proportions for 48 hours. Then the rate of cell viability was detected by MTT assay, and the median effective concentration (EC50) and combination index were calculated. Results Baicalin and taurine in 2∶1 and 1∶1 proportion groups can significantly improve learning and memory ability in AD rats (P<0.05,P<0.01). The EC50 were 2.110 μmol/L and 0.422 μmol/L, respectively. The combination index was 0.308.Conclusion Baicalin combined with taurine exhibit synergetic protective effects in AD rats induced by Aβ1-42. The optimal compatibility proportion of baicalin and taurine is 2∶1, and EC50 is 0.279 μmol/L.

OBJECTIVE: To investigate the clinical prognostic impact factors of adult sinonasal rhabdomyosarcoma (SNRMS). METHOD: The clinical features, treatment methods, and disease outcome were reviewed retrospectively for twenty-three adult SNRMS between 2006 January and 2014 December. The survival analysis was performed by Kaplan-Meier estimate and the comparison between groups by Log-rank test. Multivariate analysis was carried out by Cox proportional hazard model. RESULT: Patients' ages ranged from 18 to 59 years (median, 23.2 years). With a median follow-up of 20 moths (3-47 moths), 14 cases dead and 9 cases alive, the 1-year and 2-year overall survival (OS) rates were 77.1% and 35.0%, respectively. Within the 1-year and 2-year OS rates,early stage group had a higher overall survival rates than advanced diease group (100.0%, 66.7% and 83.3%, 10.5%, P < 0.05); combined therapy group had a higher overall survival rates than single treatment group (86.7%, 50.0% and 50.8%, 0, P < 0.05). In the non-metastasis group (21 cases), 1-year and 2-year distant metastasis rates were 38.1% and 70.5%, respectively. Multivariate analysis showed that radiotherapy, chemotherapy and tumor diameter less than 5 cm were good prognostic factors (P < 0.05), while the lymph node metastasis, meningeal involvement and orbital involvement were poor prognostic factors (P < 0 05). In the 14 cases of dead patients, 92 8% (13/14) died of distant metastasis. CONCLUSION Adult RMS had a high advanced rate with poor prognosis. Distant metastasis is the leading cause of death. Controlling distant metastasis is a key to improve the survival rate.
Adolescent ; Adult ; Humans ; Lymphatic Metastasis ; Middle Aged ; Multivariate Analysis ; Paranasal Sinus Neoplasms ; diagnosis ; mortality ; pathology ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Rhabdomyosarcoma ; diagnosis ; mortality ; pathology ; Survival Analysis ; Survival Rate ; Treatment Outcome ; Young Adult

Seven target compounds coupled by rhein and furoxan were synthesized and their chemical structures were confirmed by 1H NMR, IR, and MS. All target compounds were evaluated for anti-proliferative activity against human hepatoma cells HepG2 and Bel-7402, human colon cancer cells HCT116, human osteosarcoma cells U2OS, drug-resistant cells Bel-7402/5-FU and normal hepatocytes cells LO2 in vitro by thiazolyl blue(MTT) colorimetry. The results indicated that all target compounds had more potent anti-proliferative activity than their parent compound rhein. Additionally, compound 4g had stronger proliferation inhibitory activity on HepG2, Bel-7402, U2OS and Bel-7402/5-FU,with little effect on the proliferation of normal cells, exhibiting selective inhibitory activity. Griess assay was used to measure the release of nitric oxide in vitro. Results showed that compound 4g could increase the releases NO in HepG2 cells, which may be associated with its antitumor effects. Furthermore, the antitumor activity of compound 4g was attenuated by NO scavenger (hemoglobin), which indicates that the antitumor activity of compound 4g may be partly related to the release of NO.

Objective To observe the effect of Compoud Qingqin Liquids on renal function of rat model of uric acid nephropathy, and to discuss its protection of renal function. Methods The rat model was induced by gavaging adenine and feeding yeast. SD rats were randomly divided into control group, model group, positive group, and high-, medium-, low-dose groups of Chinese medicine. Blank control group and model group were daily gavaged with distilled water, positive control group was daily gavaged with allopurinol by 9.33 mg/kg, and high-, medium-, low-dose group of Chinese medicine was daily gavaged with Compound Qinggin Liguids by 3.77, 1.89, 0.09 g/(kg·d) respectively for 6 weeks. General condition of rats were observed, renal pathological changes were observed with light and electron microscope. Urine protein concentration, blood uric acid, blood urea nitrogen, creatinine and kidney weight index were respectively tested before and after treatment. Results There were no significant differences in eating, drinking and body weight between before and after modeling. Compoud Qingqin Liquids can obviously decrease the concentration of urine protein, blood uric acid, serum creatinine, blood urea nitrogen, and kidney weight index (P<0.05) of rats with uric acid nephropathy. Renal tubular epithelial cells atrophy and renal interstitial fibrosis of high-dose group of Chinese medicine were not evident. Conclusion Compoud Qingqin Liquids can protect the rats renal function against uric acid renal injury.