Objective To construct a T-bet response reporter gene, for the detection of T-bet tran-scriptional activity and application in high-throughput screening for the functional genomies. Methods The cis-acting DNA element, ThRE, based on CNS-22 T box site of IFN-γ gene, was recombined into a reporter vector pLUC-MCS. The reporter gene was transfected into HEK 293T cells to detect its response to T-bet. And the binding of T-bot to TbRE was identified with electrophoretic mobility shift assay(EMSA). Results ThBE was successfully cloned into pLUC-MCS, named as TbRE-LUC. Using a luciferase assay, expression of the reporter gene is found to be induced by T-bet in a dose dependent manner and correlate with T-bet ex-pression positively with activation up to 20 folds. Moreover, the binding specificity of T-bet to TbRE is vali-dated by EMSA. Conclusion We successfully constructed a T-bet response reporter gene, ThRE-LUC, which responds to T-bet keenly and specifically. TbRE-LUC will be a useful tool in high-throughput screen-ing for human genes associated with transcription activity of T-bet.

BACKGROUND: Differentiation of neural stem calls (NSCs) was mediated by many environmental factors. Several factors can induce NSCs to differentiate into neurons in varying degrees and it is now a focus on the control of NSCs differentiation.OBJECTIVE: To study the effects of combination of basic fibroblast growth factor (bFGF) and brain-derived neurotrophic growth factor (BDNF) on the differentiation of NSCs into neurons.DESIGN, TIME AND SETTING: The in vitro cytology observation was performed at the Neurotomia Laboratory of China Medical University in May 2008.MATERIALS: Three adult male SD rats were provided by Experimental Animal Center of China Medical University.METHODS: Dispositions to the rats were consistent with ethical standards of animals. The rat brain hippocampus was removed sterilely. After trypsin digestion, NSCs were cultured in serum-free medium. Cell suspension was prepared and diluted when the diameter of the fourth passage of clone sphere was 200 μm by mixture of DMEM/F12 containing 2% B27, 20 μg/L of epidermal growth factor (EGF), and 20 μg/L bFGF. Monoclonal calls were passagad. NSCs were divided into blank control, bFGF, BDNF and bFGF+BDNF groups by different growth factors added into the media. Fetal bovine serum of 0.1 volume fraction was added in blank control group. The media in the other three groups were added bFGF, BDNF and bFGF+BDNF respectively for 1 week.The concentration of bFGF was 10 μg/L and the concentration of BDNF was 200 μg/L.MAIN OUTCOME MEASURES: Immunocytochemistry staining was used to identify NSCs as well as to detect the differentiation of NSCs into neurons.RESULTS: The monoclonal calls expressed nestin and the differentiated call expressed neuron specific enolase and glial fibrillary acidic protein. Compared to blank control group, the proportion NSCs into neurons in the bFGF group, BDNF group and bFGF+BDNF group were much higher (t=3.409-7.558, P < 0.05), with the highest in bFGF+BDNF group (t =7.558, P < 0.05).CONCLUSION: Combination of bFGF and BDNF can promote the differentiation of adult hippocampus NSCs into neurons.

The paper briefed characteristics of emergency medical centers in developed countries, described the current status in China, and analyzed problems found in the emergency medical rescue system in the country.On such basis, the authors proposed on the organization, network building, communication and information system, rescue teams, rescue equipments, synergy mechanism, and assurance measures, for the purposes of capacity building and resue capacity of the pre-hospital care institutions in China.

Objective:To compare the effects of intravenous anesthesia between propofol and etomidate in patients undergoing laparoscopic surgery and their effects on plasma nitric oxide (NO) and endothelin-1 (ET-1).Methods:The clinical data of 80 patients with laparoscopic surgery in Guangrao People′s Hospital from March 2017 to March 2019 were retrospectively analyzed. Among them, 40 cases were given propofol intravenous anesthesia (propofol group), and 40 cases were given etomidate intravenous anesthesia (etomidate group). The anesthetic effect, plasma NO and ET-1 levels, hemodynamic indexes and adverse reactions (muscle spasm, nausea and vomiting, injection site pain, body movement and respiratory depression) were compared between the two groups.Results:The time of consciousness disappearance, tracheal intubation, eye opening, spontaneous breathing and speech response in etomidate group were significantly shorter than those in propofol group: (57.48 ± 2.63) s vs. (86.17 ± 7.41) s, (4.39 ± 2.56) min vs. (6.42 ± 2.58) min, (5.39 ± 2.56) min vs. (9.42 ± 2.58) min, (5.21 ± 1.99) min vs. (8.75 ± 2.54) min and (8.39 ± 2.56) min vs. (8.39 ± 2.56) min, and the differences were statistically significant ( P<0.05). The levels of NO and ET-1 in the etomidate group were significantly lower than those in the propofol group at 0.5, 1.0 and 1.5 h after pneumoperitoneum ( P< 0.05). The levels of systolic blood pressure, diastolic blood pressure and oxygen saturation (SpO 2) in the etomidate group were significantly higher than those in the propofol group: (78.42 ± 4.68) mmHg (1 mmHg = 0.133 kPa) vs. (74.11 ± 6.63) mmHg, (132.86 ± 8.71) mmHg vs. (111.24 ± 3.56) mmHg and 0.982 ± 0.032 vs. 0.953 ± 0.043, and the differences were statistically significant ( P<0.05). The incidence of adverse reactions in the etomidate group was significantly lower than that in the propofol group: 17.5% (7/40) vs. 47.5% (19/40), P<0.05. Conclusions:Compared with propofol intravenous anesthesia, etomidate intravenous anesthesia in laparoscopic surgery patients has more stable hemodynamics and better anesthetic effect. It can effectively inhibit the release of NO and ET-1, and has higher safety.

Objective:The application of computer-rapid prototyping-assisted simulated hollow screw in the treatment of femoral neck fractures in fresh cadavers verified the reliability and accuracy of computer-rapid prototyping-assisted simulated hollow screw in the treatment of femoral neck fractures.Methods:The glass ion was labeled as a target for the treatment of femoral neck fracture with a cannulated screw. Three Kirschner pins were placed at the top and bottom of the greater trochanter. CT scan of the processed specimens. The scan data was imported into Mimics software, and the upper end of the femur was reconstructed three-dimensionally and the target of the femoral neck fracture was fixed with cannulated screws. The position of the cannulated screw in the femoral neck was designed. In Pro/E software, draw a guide that fits the three positioning Kirschner pins and cannulated screw guide needles on the upper end of the femur, and use rapid prototyping technology to make the guide. The navigation template was mounted on the positioning pin through the cannulated screw guiding hole, and the hollow screw guiding needle was drilled, and the hollow screw was inserted for the treatment of the femoral neck fracture to complete the positioning and guiding. According to the coordinates of the hollow screw guide needle and the target center in the Mimics software three-dimensional reconstruction coordinate system, the spatial distance calculation formula is used to calculate the distance between the hollow screw guide needle and the target center.Results:The coordinate analysis of the end point of the cannulated screw guide needle and the target can be obtained that the average distance between the end point of the cannulated screw guide needle for fixed femoral neck fracture and the target center point was 1.92 mm.Conclusion:The computer-rapid molding-assisted cannulated screw fixation of the femoral neck fracture is reliable and accurate, which provides a new idea and method for clinical individualization and precise fixation of the femoral neck fracture.

Objective To investigate the efficacy of biphasic insulin aspart 50(BIAsp50)twice daily(bid) versusbiphasichumaninsulin50(BHI50)(bid)plusmetforminonbloodglucosecontrolfollowingastandardmealtest in Chinese patients with type 2 diabetes mellitus(T2DM). Methods A randomized, open-label, 2-sequence, crossover trial for two 4-week treatment periods was conducted in 14 Chines institutes. Eligible subjects inadequately controlled with BHI50(bid)plus metformin were randomized to two sequences in a 1 : 1 ratio(A:BIAsp50-BHI50, B:BHI50-BIAsp50 ) . Standard meal tests were performed at baseline and the ends of two periods within 4 weeks. Primary endpoint was 2h postprandial plasma glucose ( PPG) increment following standard meal test, with insulin dose standardized at 0. 3 IU/kg. Results A total of 161 subjects were randomized into two sequences(81 to sequence A, and 80 to sequence B) and finally analysed. After 4 weeks of treatment, mean 2h PPG increment with BIAsp50 was lower than that with BHI50 [ treatment difference of BIAsp50 vs BHI50: -1. 12 mmol/L ( 95% CI-1. 66,-0. 58), P<0. 01], suggesting superiority of BIAsp50 over BHI50. Incremental area under the curve for PPG(0-2 h)with BIAsp50 was lower than that with BHI50 [treatment difference:-38. 8 mmol·L-1·min-1(95%CI-77. 3,-0. 26), P=0. 049], as was the mean 2h PPG [treatment difference:-0. 58 mmol/L(95% CI -1. 13,-0. 03), P=0. 040]. The FPG value with BIAsp50 was higher than that with BHI50 [treatment difference:0. 52 mmol/L(95%CI 0. 18, 0. 86), P=0. 003]. The rate of nocturnal hypoglycemia with BIAsp50 was lower than that with BHI50(1. 13 vs 2. 86 events per subject year, P<0. 01). Conclusion In patients with T2DM inadequately controlled with BHI50 plus metformin, BIAsp50 was proven to be well-tolerated with improved postprandial glucose control compared with BHI50.

Objective To evaluate the efficacy and safety of epalrestat, an aldose reductase inhibitor, and epalrestat plus methylcobalamine on diabetic peripheral neuropathy, as compared with methylcobalamine. Methods A total of 444 subjects with diabetic neuropathy were enrolled in the study, and divided into methylcobalamine group ( n= 145 ) , epalrestat group ( n = 143 ) , and methylcobalamine combined with epalrestat group ( n = 156 ) . Therapeutic efficacay was assessed in terms of clinical symptoms and physical examinations by using Michigan Neuropathy Screening Instrument ( MNSI ) , and electrophysiological assessments. Results After 4 to 12-weeks′treatment, symptoms and signs of neuropathy ( using MNSI ) are significantly improved in the three groups ( P<0. 01). The mean changes of MNSI ( questionnaire) score from baseline were higher in epalrestat group and methylcobalamine combined with epalrestat group as compared with that of methylcobalamine group(P<0. 05), but no difference was detected in the change of MNSI ( physical examination ) score from baseline among three groups. After treatment for 12 weeks, motor nerve conduction velocity ( MNCV ) was significantly improved in epalrestat group and methylcobalamine combined with epalrestat group(P<0. 05), but no difference was detected in MNCV at 12 week among three groups(P>0. 05). Conclusion Epalrestat is effective and safe in the treatment of diabetic neuropathy. Furthermore, epalrestat is more efficacious in ameliorating symptoms and MNCV of neuropathy than methylcobalamine. However, while no improved efficacy is shown with the combined treatment.

Impaired eady phase insulin secretion is an important reason for leading to postprandial hyperglycemia.Nateglinide is a rapid-acting insulin secretagogue,which reduces postprandial blood glucose of type 2diabetic patient by restoring early phase insulin secretion.The efficacy and safety have been fully verified by clinical administration and it is more widely used to treat type 2 diabetic patients.Both sulfonylureas and glinides were named insulin secretagogue agents and regarded as alternative first-line drugs in the 2010 Chinese Guideline for treatment of type 2 diabetes.AACE/ACE Consensus statement claimed that glinides would be one of the important choices after metformin.In order to further guide the clinical application of nateglinide,16 national specialists in the field of endocrinology and metabolism of China discussed,drafted,and edited this consensus.The current consensus combined clinical evidences at home and abroad.systematically reviewed and summarized tlle results of these studies about nateglinide.It will provide guiding recommendations and reference concerning how to reasonably and effectively use nateglinide in the clinical practice.

Oblique lateral interbody fusion (OLIF) was minimally invasiveprocedure for lumbar interbody fusion (LIF) through the space between anterior margin of retroperitoneal psoas major muscle and the vessels (ATP). Although OLIF had many advantages over other approaches, there were also various kinds of surgical complications, the incidence of which was 3.69%-81.82%. Most of the complications were relieved by conservative or symptomatic treatment. Only a small number of complications were difficult to recover, if so, revision surgery was needed and might remain persistent symptoms. OLIF complications included intraoperative and postoperative complications. Major vascular injury was a dangerous complication during operation, which requires immediate compression or suture to prevent bleeding. The incidence of nerve injury could be reduced by avoiding violent traction and tissue separation and reducing the operation time. When injury of thorax and peritoneum occurs, suture should be done as soon as possible. Transient hip flexion weakness and transient thigh/groin sensory disturbance was the most common post-operative complication, most of which would disappear after several months. Intestinal obstruction is caused by the pulling of peritoneum during operation, most of which was incomplete and would be relieved after some time. Postoperative infection was mostly superficial and would be cured by dressing change and antibiotic application. Subsidence of cage and collapse of intervertebral space were the most common complications related to instrumentations which might not lead to related clinical symptoms; however the severe cases need to be repaired. The incidence of pseudarthrosis is relatively low and a few patients with clinical symptoms need revision surgery. The sample size of most studies was small and follow-up period was short. In the future, large samples and multi-center studies are needed to improve our understanding of OLIF complications in the future.

Objective:To investigate the effect of O-arm navigation assisted posterior pedicle screw reduction and internal fixation of atlantoaxial fractures.Methods:A retrospective case-control study was conducted to analyze 37 patients with atlantoaxial fractures admitted to Third Hospital of Hebei Medical University from January 2016 to June 2018, including 22 males and 15 females, aged from 29 to 68 years [(50.9±9.8)years]. The posterior pedicle screw reduction and internal fixation was performed under O-arm navigation system (navigation group, n=24), and using free-hand technique (free-hand group, n=13). The operation time and blood loss were compared between the two groups. The Japanese Orthopaedic Association (JOA) score and neck disability index (NDI) were used to evaluate the clinical efficacy before operation, 7 days operation and at the last follow-up. The complications were detected. A total of 86 screws were placed in navigation group (Neo grade 0: 83 screws, grade 1: 2 screws, grade 2: 1 screw ), and 44 screws were inserted in free-hand group (Neo grade 0: 36 screws, grade 1: 5 screws, grade 2: 2 screws, grade 3: 1 screw)( P<0.05). Classification of screw positions proposed by Neo et al was used to evaluate the position relationship between the screw and the bone cortex and the incidence of screw penetration. Results:All patients were followed up 24-38 months [(27.7±4.0)months]. The operation time in navigation group was (189.8±35.4)minutes, significantly shorter than (221.5±48.6)minutes in free-hand group ( P<0.05). The bleeding volume in navigation group was 300.0 (250.0, 537.5)ml , significantly less than 500.0 (425.0, 625.0)ml in free-hand group ( P<0.05). Both groups showed significantly enhanced JOA and decreased NDI after operation and at last follow-up, compared with those before operation ( P<0.05). However, there was no significant difference in JOA and NDI between the two groups ( P>0.05). No severe complications such as neurovascular injury occurred during operation. The incidence of cortical penetration was 3% (3/86) in navigation group and 18% (8/44) in free-hand group ( P<0.05). Conclusions:In the process of posterior atlantoaxial pedicle screw placement, the application of O-arm navigation can significantly reduce the operation time and amount of bleeding, and enhance the accuracy of pedicle screw implantation.