Glioma-associated microglial cells, a key component of the tumor microenvironment, play an important role in glioma progression. In this study, the mouse glioma cell line GL261 and the mouse microglia cell line BV2 were chosen. First, circadian gene expression in glioma cells co-cultured with either M1 or M2 microglia was assessed and the exosomes of M2-polarized and unpolarized BV-2 microglia were extracted. Subsequently, we labeled the exosomes with PKH67 and treated GL261 cells with them to investigate the exosome distribution. GL261 cell phenotypes and related protein expression were used to explore the role of M2 microglial exosomes in gliomas. Then a specific miR-7239-3p inhibitor was added to verify miR-7239-3p functions. Finally, the mouse subcutaneous tumorigenic model was used to verify the tumorigenic effect of M2 microglial exosomes in vivo. Our results showed that in gliomas co-cultured with M2 microglia, the expression of the BMAL1 protein was decreased (P < 0.01), while the expression of the CLOCK protein was increased (P < 0.05); opposite results were obtained in gliomas co-cultured with M1 microglia. After treatment with M2 microglial exosomes, the apoptosis of GL261 cells decreased (P < 0.001), while the viability, proliferation, and migration of GL261 cells increased. Increased expression of N-cadherin and Vimentin, and decreased E-cadherin expression occurred upon treatment with M2 microglial exosomes. Addition of an miR-7239-3p inhibitor to M2 microglial exosomes reversed these results. In summary, we found that miR-7239-3p in the glioma microenvironment is recruited to glioma cells by exosomes and inhibits Bmal1 expression. M2 microglial exosomes promote the proliferation and migration of gliomas by regulating tumor-related protein expression and reducing apoptosis.

OBJECTIVE：To pr ovide reference for improving extended clinical trial system in China. METHODS ：The implementation experience of the international extended clinical trial system was introduced from three aspects ： system development，application scope and principles ，application and approval ；the evolution and challenges of the system in China were further analyzed ，and relative suggestions were put forward. RESULTS & CONCLUSIONS ：The development of international extended clinical trial system in the United States ，the United Kingdom ，Australia and the European Union had been relatively mature. The specific system had not formally been established in China ，and the implementation of the system faced many challenges，such as the ethical problems caused by drug use risk ，the sponsor dilemma caused by the difficult balance between the resistance of initiating application and the benefits ，the difficulty of review caused by the unknown responsible party of risk assessment and the urgent time of approval. It can be dealt with by the following measures ：improving the existing laws and regulations，strengthen information disclosure and risk prevention work ，make clear the division of responsibilities in the process of system operation. Meanwhile ，based on the existing international model ，the management system of the expanded clinical trials in China is established. Corresponding application paths are set up for different application types such as “single patient emergency ”， “single patient non emergency ”and“two or more patients group ”，and all parties should be mobilized to supervise so as to promote the improvement and implementation of the extended clinical trial system.

Objective:To explore the application value in cerebral blood perfusion status of dynamic susceptibility contrast enhanced perfusion weighted imaging (DSC-PWI) in ischemic cerebrovascular disease (ICVD).Methods:Retrospective analysis of 31 cases ICVD head magnetic resonance imaging (MRI), diffusion weighted imaging (DWI), magnetic resonance angiography (MRA) and DSC-PWI image in clinical diagnosis, and selectively analyze parameters including relative cerebral blood flow (rCBF), relative cerebral blood volume (rCBV), mean transit time (MTT) and time to peak (TTP) for generalize its characteristics.Results:31 cases of ICVD, 16 cases hypoperfusion, rCBF decreased significantly, rCBV decreased, MTT and TTP prolonged, include 7 cases of large vessel disease and 5 cases of small vessel disease. The collateral circulation formed 9 cases, including 3 cases with good compensation, rCBF normal, rCBV normal, MTT and TTP prolonged, 6 cases with bad compensation, rCBF decreased, rCBV normal or increased, MTT and TTP prolonged. The blood reperfusion in 3 cases, rCBF normal or slightly increased, rCBV increased, MTT shortened or normal, and TTP shortened. The excessive perfusion in 3 cases, rCBF significantly increased and rCBV significantly increased, with MTT and TTP shortened.Conclusions:DSC-PWI can reliably reflect the perfusion state and collateral circulation compensation of ICVD, so as to guide the selection of clinical treatment program and significantly improve the prognosis of patients.

OBJECTIVE： To provide reference for the thought and operation of the law to protect the tacit technology secrets of traditional Chinese medicine （TCM）. METHODS： Through the analysis of the composition， categories and legal attributes of tacit technology secrets of TCM， legal protection methods were sought from the legal perspective. RESULTS & CONCLUSIONS： In the field of TCM， tacit knowledge， represented by the TCM processing， TCM diagnosis and treatment， is mostly inherited and studied in the form of technology secrets. The tacit technology secrets of TCM have the characteristics of personality dependence and inheritance， the protection of them by law can not be achieved directly through the field of intellectual property rights， but through the protection of the interests of the inheritors. Public law and private law are indispensable to the protection of tacit technology secrets of TCM. Public law should focus on improving the mechanism of technological inheritance and protecting the rights and interests of TCM inheritors. Private law should overcome the risk of infringement by signing contracts and transforming tacit knowledge of TCM.

OBJECTIVE:To provide reference for improving legal protection system of proprietary technology in TCM time-honored enterprises. METHODS:By studying the relevant literatures and data,main type of proprietary technology,formula, legal characteristics of processing technology in TCM time-honored enterprises were analyzed;the protection forms and existing problems of proprietary technology were summarized to put forward countermeasures. RESULTS & CONCLUSIONS:The most important proprietary technology of TCM time-honored brand is formula and TCM processing technology,and legal characteristics of it include practicality,value,confidentiality and teachability. The proprietary technology protection of TCM time-honored brand included various methods such as business secret,patent protection and administrative protection. But,there still are problems such as the incompatibility of patent protection,the conflict between the secret protection and information disclosure,gradually weakened means of administrative protection,weak awareness of legal protection in time-honored brand of TCM. It is suggested to use comprehensive protection means of patent system and business secret,coordinate intellectual property protection and administrative protection,establish proprietary technology protection directories and databases and incentive and constraint mechanism of internal intellectual property rights in enterprises so as to seek effective way to protect proprietary technology of TCM time-honored brand.

Objective To promote the application of the best evidence for supportive care in the management of depression in patients with head and neck cancer, and to improve the quality of clinical care.Methods Follow the three stages of clinical evidence practice of Joanna Briggs Institute (JBI): baseline review, practice change and effect evaluation, the best evidence of supportive nursing in depression management for head and neck cancer patients were retrieved, and 4 quality review indicators were developed. A total of 50 hospitalized head and neck cancer patients and 12 nurses were selected to collect data by means of field observation, interviews and nursing records. The obstacles to the application of evidence were analyzed and a practical scheme was worked out.Results The nurse's implementation rate of the four review indicators increased from 0-90％ at the baseline review to 90％-100％ after the evidence was applied. Except for Indicator 3 "Indicators nurses introduced the pressure on patients and their families to face cancer", the implementation rates of the other three indicators were significantly improved (P<0.01). After the application of evidence, the incidence of depression was decreased (P<0.05). The score of Relaxation, Sleep and Fatigue Management Knowledge Questionnaire was (18.20±14.09) before the application of evidence, which was lower than that after the application of evidence (80.60±12.84), and the difference was statistically significant (t=-23.14, P<0.01). The nurses' score of Supportive Nursing Knowledge Questionnaire before the application of evidence was (63.83±10.78), which was lower than that after the application of evidence (95.5±3.18), and the difference was statistically significant (t=-9.76,P<0.01).Conclusions The evidence-based application program can standardize nurses' supportive nursing behaviors for patients with depressive symptoms, reduce the severity of depression, improve the self-support knowledge level, and improve the quality of depression management in hospitalized patients with head and neck cancer.

Objective:To evaluate the feasibility of transforaminal endoscopic nerve root decompression for degenerative lumbar spinal stenosis (DLSS).Methods: From July 2011 to April 2016,96 cases of single segment DLSS were involved.All the patients had unilateral lower extremity neurological symptoms,signs,neurogenic intermittent claudication of less than 500 m.Imaging examinations (CT or MRI) or diagnostic nerve root block confirmed single segment degeneration.The mean age was (71.6±5.4) years,male: 55 cases,female: 41 cases.Their intraoperative blood loss,operation time,complications,ambulation time and discharge time were recorded.Leg pain VAS,ODI were used to evaluate the pain and lumbar function of the patients.The clinical efficacy was evaluated by Nakai evaluation.Results: All the patients were performed endoscopic decompression of the lateral recess and nerve root by removing the ventral part of the superior facet joint,the ligamentum flavum and the intervertebral disc.The decompression range was from the inferior edge of the upper pedicle to the superior edge of the lower pedicle.The nerve root was detected to have no compression and the pulse of nerve root returned to normal.The patient got ambulant on the operation day and discharged if he had no discomfort symptom.In the study,68 cases got follow up.The mean follow-up time was 12.1 months (6-63 months).The VAS at dif-ferent follow-up time points was improved relative to the baseline,and the difference was statistically significant (F=491.60,P<0.001).The ODI at different follow-up time points was improved relative to the baseline,and the difference was statistically significant (F=189.91,P<0.001).The excellent and good rates of Nakai evaluation were 79.4％ (excellent in 42 cases,good in 12 cases,fair in 10 cases and poor in 4 cases).The mean intraoperative blood loss was (49.29±11.86) mL.The mean operation time was (92.46±21.34) min.The mean ambulation time was 1.8 h.The mean discharge time was 2.3 days.Postoperative epidural hematoma was found in 1 case.Foot drop was found in 1 case.Second stage open surgery was performed in 6 cases.Conclusion: We can apply transforaminal endoscopic decompression for the patients of lumbar spinal stenosis who have unilateral nerve root irritation.Patients with transforaminal endoscopic decompression can get less surgical trauma,quick recovery and obtain good short-term outcome.

This study aimed at investigating the dose-effect relationship of SFI between the blood viscosity and the early-and mid-stage cardiogenic shock and the mediatory effect on rats.The end or root of left anterior descending coronary arteries (LADCA) was ligatured to establish the rat model of the early-and mid-stage cardiogenic shock.The blood viscosity indexes included low shear rate (LSR,10/s),middle shear rate (MSR,60/s),high shear rate (HSR,150/s) of the whole blood viscosity and plasma viscosity (PV),being observed 60 mins after the venous administration of 0.10,0.33,1.00,3.30,10.00 and 20.00 mL·kg-1 SFI (low dosage range:0.1-1.0 mL·kg-1,middle dosage range:1.0-10 mL·kg-1,high dosage range:10-20 mL·kg-1) with a blood rheometer.Dose-response curves were fitted by GraphPad Prism 6.0 software,the dose-response relationship of SFI between the blood viscosity and the early-and mid-stage cardiogenic shock in rats was evaluated to calculate the dose threshold parameters of the indexes.It was found that the blood viscosity indexes were improved with the dosage of 10 mL·kg-1 SFI in rats with the early-stage cardiogenic shock,while the dose-response curves of LSR,MSR and HSR at the early stage all presented favorable s shapes.Most of the effective dose range [D]2o-[D]80 and the threshold dose [D]20 were between 3.3 and 6.3 mL· kg-1.The four indexes of blood viscosity were improved with the administration of 10 and 20 mL·kg-1 SFI in mid-stage model rats with favorable s shapes in the dose-response curves.Most of the effective dose range and the threshold dose were in the range of 3.3 to 10.0 mL·kg-1.In conclusion,most of the dose-response curves of blood viscosity indexes in early-and mid-stage cardiogenic shock rats presented favorable s shapes with the threshold dose between 3.3 and 10.0 mL·kg-1,indicating an effective middle dosage range,which was converted into clinical dosage about 37.1 to 112 mL each day.The research provided an experimental basis for clinical medication.

Objective To evaluate the clinical use of delayed open reduction and internal fixation technique in the treatment of Rud?i Ⅲ Pilon fractures. Methods From May 1997 to June 2003, 38 patients with Rud?i Ⅲ Pilon fractures underwent immediate calcaneal traction as well as treatment with methylprednisolone and mannitol. After 5-8 days, all patients underwent regular open reduction and internal fixation with cloverleaf buttress plates. One week after the operation, ankle joint began to exercise with no weight bearing. Results All patients were followed up for average 18 months and evaluated subjectively and objectively by using systems of Teeny and Wiss, which showed good results for 74 %, mild for 21% and poor for 5%. All fractures healed within an average of 5.7 months except for two cases of infections and one soft tissue complication. Conclusion Delayed open reduction and internal fixation offers acceptable way for treatment of severe Pilon fractures.

Objective To study the application of extended tibia tubercle osteotomy in revision of total knee arthroplasty. Methods Retrospective study was carried out in 12 cases that had received extended tibia tubercle osteotomy in Zhongshan Hospital from May 1998 to October 2002. Results After osteotomy, all knees were healed clinically and radiologically at 6th month postoperatively. Average range of joint motion increased from 79 degrees to 95 degrees. Average knee score improved from 60 to 85. Conclusion Extended tibia tubercle osteotomy can improve the exposure of operation and knee joint function.