Objective To establish a RP-HPLC method for the determination of encapsulation efficiency (EE) and medicine loading (ML) in β-elemene-loaded Nano Polymeric Micelles; To study its release characteristic in vitro. Methods DSPE-PEG2000 was used as carrier to prepare medicine-loaded micelles. EE and ML were determined by petroleum ether extraction method, and its release characteristic in vitro was studied by dialysis method. Results The average EE and ML ofβ-elemene-loaded Nano Polymeric Micelles were 89.47%and 8.33%, respectively. Its release characteristic was slow. Conclusion The method for EE and ML determination is simple and accurate, and the prepared micelles have the property of sustained release.

Objective: To select suitable conditions for prokaryotic expression and purification of rhIL-17. Methods: rhIL-17 was expressed in E. coli host under heat induction. After compared among the expression amounts in different media under different heat induction time, the most suitable conditions was selected. The target protein was present in the form of inclusion body. The precipitate of inclusion was obtained and purified after 6M guanidine solublization or 2% SDS solublization. Results: Either protocol could yield rhIL-17 with high purity and stable activity. The SDS solublization mehthod gives rise to much more higher productivity than the guanidine solublization method. Conclusion: rhIL-17 were expression in E. coli system and purified to homogenicity by SDS solublization methods with high productivity.

Objective To evaluate the role of β-arrestin-1 in penehyclidine hydrochloride (PHC)-induced inhibition of lipopolysaccharide (LPS)-caused increase in pulmonary microvascular permeability in human pulmonary microvascular endothelial cells (PMVECs).Methods Human PMVECs were seeded in 6-well plates (2 ml/well) or in culture flasks (4 ml/flask) at the density of 1 × 105 cells/ml and divided into 5 groups (n=15 each) using a random number table:empty plasmid transfection group (group C),LPS plus empty plasmid transfection group (LPS group),PHC plus LPS plus empty plasmid transfection group (P+LPS group),LPS plus β-arrestin-1 short hairpin RNA (shRNA) transfection group (LPS+shRNA group) and PHC plus LPS plus β-arrestin-1 shRNA transfection group (P+LPS+shRNA group).In LPS and LPS+shRNA groups,the cells were transfected with empty plasmid 1.5 μg or with plasmid containing 15 nmol/L β-arrestin-1 shRNA,LPS with the final concentration of 0.1 μg/ml was added at 24 h of incubation,and the cells were then incubated for 1 h.In P+LPS and P+LPS+shRNA groups,the cells were transfected with empty plasmid 1.5 μg or with plasmid containing 15 nmol/L β-arrestin-1 shRNA,PHC with the final concentration of 2 μg/ml was added at 24 h of incubation,LPS with the final concentration of 0.1 μg/ml was added at 1 h of incubation,and the cells were then incubated for 1 h.The cell permeability was measured using Transwell chambers.The expression of heat shock protein (HSP27) was detected by immunofluorescence.The expression of β-arrestin-1,p38 mitogen-activated protein kinase (p38MAPK) and phosphorylated p38MAPK (p-p38MAPK) was detected by Western blot.The ratio of pp38MAPK/p38MAPK was calculated.Results Compared with group C,the cell permeability was significantly increased,the expression of HSP27 was up-regulated,p-p38MAPK/p38MAPK ratio was increased,and the expression of β-arrestin-1 was down-regulated in LPS,LPS + shRNA and P + LPS + shRNA groups (P＜0.05),and no significant change was found in the parameters mentioned above in group P+LPS (P＞ 0.05).Compared with group LPS,the cell permeability was significantly decreased,the expression of HSP27 was down-regulated,p-p38MAPK/p38MAPK ratio was decreased,and the expression of β-arrestin1 was up-regulated in group P +LPS,and p-p38MAPK/p38MAPK ratio was significantly increased (P＜0.05),and no significant change was found in the other parameters in group P+LPS+shRNA (P＞0.05).Compared with group P+LPS,the cell permeability was significantly increased,the expression of HSP27 was up-regulated,p-p38MAPK/p38MAPK ratio was increased,and the expression of β-arrestin-1 was down-regulated in group P+LPS+shRNA (P＜0.05).Conclusion The mechanism by which PHC inhibits LPS-induced increase in pulmonary microvascular permeability is totally related to β-arrestin-1 in human PMVECs.

Adenovirus harboring E. coli. cytosine deaminase gene (AdCD) and adenovirus encoding with lymphotactin gene (AdLtn) were used for gene therapy in vivo. BALB/c mice were inoculated subcutaneously with CT26 colon adeno-carcinoma cells and 3 days later received combined injection of AdCD and/or AdLtn followed by continuous injection with 5-fluorocytosine(5-FC) 300mg/kg. The results demonstrated that mice received combined therapy developed tumors most slowly and survived longest when compared with mice treated with AdCD/5-FC, AdLtn, AdlacZ/5-FC or PBS. To further explain the immunological mechanism of the antitumor effects by the combined therapy, we found that combined treatment with suicide gene and Ltn gene therapy achieved maximal cytotoxic effects of nature killer cells and specific cy-totoxic T lymphocytes. FACS analysis of the tumor mass demonstrated that AdCD/5-FC in combination with AdLtn therapy increased the expression of H-2K~d and B7-1 expression on tumor cells. The CD4~+ and CD8~+ cells infiltrated in the tumor mass after combined therapy were significantly increased when measured by FACS analysis. Our results demonstrated that combined transfer of suicide gene and lymphotactin gene induce nonspecific and specific antitumor immunity of the host and elicit more potent antitumor effect.

Objective To investigate the feasibility and clinical effect of immediate ureterovesical reimplantation after ureteral rupture during laparoscopic hysterectomy. Methods From August 2010 to December 2015, 5 cases of ureteral rupture during the operation of laparoscopic hysterectomy were treated with immediate ureterovesical reimplantation under laparoscopy. Results All operations were successfully performed without transversion to open surgery. No patients with urinary leakage occurred. The mean follow-up were 21 months (range 3-60 months) . No cases with ureteral stricture were observed. Slight urine reflux was found in two patients, of whom obvious hydronephrosis and renal damage were not found. Conclusion Immediate ureterovesical reimplantation under laparoscopy is a feasible, safe and minimal invasive method for treatment of ureteral ruputure during laparoscopic hysterectomy.

The transcriptional repressor RE1 silencer transcription factor(NRSF/REST) is an important factor that restricts some neuronal traits in neurons.Since these traits are also present in pancreatic islet cells,NRSF-regulated genes involved in islet function are searched.A NRSE-like motif was analysed in human insulin promoter.The role of NRSE was evaluated by generating a model of insulin-secreting cells that firmly express NRSF.The presence of NRSF led to a decrease in activity of human insulin promoter by stable or transient transfection with human insulin-promoter luciferase.The predicted NRSE-like motif also confers NRSF-dependent transcriptional repression in the context of a surrogate gene promoter.Specific binding activity of NRSF/REST to the NRSE-like motif was confirmed by EMSA.Moreover,the binding activity is competed by consensus NRSE sequence.These data showed that human insulin promoter is regulated by the transcriptional repressor NRSF/REST via the NRSE-like motif.

Objective To evaluate the role of M3 receptors in penehyclidine hydrochloride ( PHC)-induced reduction of lipopolysaccharide ( LPS)-induced injury to human pulmonary microvascular endotheli-al cells ( PMVECs) . Methods Human PMVECs transfected with M3 shRNA were seeded in 6-well plates (2 ml∕hole) or in culture flasks (4 ml∕flask) at the density of 1×105 cells∕ml and divided into 5 groups ( n=5 each) using a random number table method: control group ( group C) , LPS group, PHC plus LPS group ( group P+LPS) , LPS plus M3 shRNA transfection group ( group LPS+shRNA) , and PHC plus LPS plus M3 shRNA transfection group ( group P+LPS+shRNA) . Group C received no mediation, and LPS was added at the final concentration of 0. 1 μg∕ml in the other groups. PHC 2 μg∕ml was added at 1 h before adding LPS in P+LPS and P+LPS+shRNA groups. The cells were transfected with plasmid containing 2. 5 nmol∕L M3 receptor shRNA in LPS+shRNA group and P+LPS+shRNA group. Contents of filamentous actin ( F-actin) in endothelial cells were measured by flow cytometry at 1 h after adding LPS. The expression of myosin light chain kinase ( MLCK) and VE-cadherin protein was examined by immunofluorescence. The ex-pression of nuclear factor kappa B ( NF-κB) p65 and IκB was detected by Western blot. Contents of tumor necrosis factor-alpha ( TNF-α) and interleukin-6 ( IL-6) were determined by enzyme-linked immunosorbent assay. The M3 receptor mRNA transcription was detected by real-time polymerase chain reaction at 10, 30 and 60 min after adding LPS. Results Compared with group C, F-actin content was significantly de-creased, the expression of VE-cadherin and IκB was down-regulated, the contents of TNF-αand IL-6 were increased, and the expression of MLCK and NF-κB p65 was up-regulated in LPS and P+LPS groups ( P<0. 05) . Compared with group C, the expression of M3 receptor mRNA was significantly up-regulated in group LPS ( P<0. 05) , and no significant change was found in group P+LPS ( P>0. 05) . Compared with group LPS, F-actin content was significantly increased, the expression of VE-cadherin and IκB was up-reg-ulated, the contents of TNF-αand IL-6 were decreased, and the expression of MLCK, NF-κB p65 and M3 receptor mRNA was down-regulated in group P+LPS and group LPS+shRNA ( P<0. 05) . Compared with group P+LPS, F-actin content was significantly increased, the expression of VE-cadherin and IκB protein was up-regulated, TNF-α and IL-6 contents were decreased, and the expression of MLCK, NF-κB p65 and M3 receptor mRNA was down-regulated in group P+LPS+shRNA ( P<0. 05) . Conclusion PHC re-duces LPS-induced injury to human PMVECs through interfering with M3 receptors and inhibiting NF-κB-mediated inflammatory responses.

Objective To explore the surgical feasibility and clinical efficacy of one-stage anterior-posterior approaches in treatment of severe fracture and dislocation of lower cervical spine in "beach chair position".Methods Sixteen male cases of severe fracture and dislocation of lower cervical spine and with a mean age of 49.8 years (range,36-78 years) treated surgically from May 2012 to May 2016 were analyzed retrospectively by using case series study.The segment of injury was C4-5 in 4 cases,C5-6 in 7 and C6-7 in 5.The degree of spinal cord injury according to the American Spine injury Association (ASIA) score was Grade A in 4 cases,Grade B in 7 and Grade C in 5.Sub-axial injury classification (SLIC) score was 8 points in 9 cases and 9 points in 7.After a general anesthesia,a ring with a hole was hanged on patient's head before the operation.Then,under the protection of hole traction,the upper of operating bed was swung up slowly,so that the patient was restricted in vertical "beach chair position" with traction on the halo in order to immobilize the head and partially reduce the kyphotic deformity.Routine cervical anterior-posterior approach was done with the exposure of damaged section of the front and rear structure.Pedicle screw system or lateral mass screw displacement was conducted.Anterior intervertebral discectomy or fracture vertebral was performed,using collaborative reset prying method before and after the road.In the front of intervertebral cage or titanium net support bone graft,rear pedicle screws or lateral mass screws fixation and bone graft fusion were implemented.The operation time and blood loss were recorded.The healing of the wound was observed.The recovery of neurological function was evaluated according to the ASIA grade.Postoperative review X-ray,CT and MRI were done to evaluate the reset and bone graft in position and fusion.Results All the surgeries were done well without aeroembolism and other related complications.The mean operative time was 153 minutes (range,150-180 minutes),and the mean amount of blood loss was 543 ml (range,400-800 ml).Sixteen cases were followed-up from 6 to 24 months (mean 13.7 months).All the incision were healed at Ⅰ stage.Spinal cord function did not aggravate.The ASIA grade was improved with an average of one to two Grades 6 months after surgery.Postoperative X-ray and CT confirmed that graft object position was favorable and cervical sequence was recovered well.The Cobb angle decreased from (23.6 ± 5.3) ° preoperatively to (4.0 ± 0.4)°postoperatively,and the translational displacement of vertebral body was restored into (2.7 ±0.4) mm (P ＜ 0.01) from (10.9 ± 1.6) mm before operation.The cervical spinal canal was not obstructed and the cervical spinal cord was relieved,showed by MRI.Conclusions One-stage anterior-posterior approaches for severe fracture and dislocation of lower cervical spine circumferential reconstruction in "beach chair position" is a beneficial and effective method,without the need of changing positions in a collaborative reduction and fixation.The method can reduce the interference of spinal cord,shorten the operation time and save anterior extra fixation.

Objective To investigate the effect of subclinical epileptiform discharges (SED) on the cognition of adult patients with epilepsy,exploring the mechanism of SED that leads to cognitive impairment in adult patients with epilepsy to raise physicians' attention about SED.Methods Patients were collected in the Department of Neurology,China-Japan Union Hospital of Jilin University from March 2016 to March 2017.Sixty adult patients with epilepsy without clinical episodes in the last three months were selected as SED group and 40 healthy volunteers as control group.Medical history of the SED group was recorded in detail.All patients were examined by Self-rating Anxiety Scale,Self-rating Depression Scale,Pittsburgh Sleep Quality Index scale in order to exclude organic brain disorders,metabolic diseases,anxiety,depression,sleep disorders and drug-induced cognitive dysfunction.Subjects in the two groups received Montreal Cognitive Assessment (MoCA),electroencephalogram and blood oxygen level dependent functional magnetic resonance imaging examination.Finally,the results were compared between the two groups.Results ①SED had different effects on cognitive function in adult patients with epilepsy,and the MoCA score (26(22,27)) showed statistically significant difference compared with the control group (29 (28,29),Z =-6.26,P =0.00).②Different discharges indexes showed different effects on cognitive function aspects.Cognitive impairment was significant when the discharges indexes were ＞ 10％ (discharges indexes 1％-10％:MoCA score 26(26,28),discharges indexes 10％-50％:MoCA score 22(19.5,25),Z =-4.74,P =0.00).③The cognitive function of epilepsy patients was positively correlated with the duration of education (r =0.41,P =0.00) and the time interval to recent seizure (r =0.31,P =0.02),and negatively correlated with SED (r =-0.57,P=0.17).There was no correlation between cognitive function and duration of disease and onset age.The SED was the main influencing factor of cognitive function in epilepsy.④Compared with healthy people,epilepsy patients with SED showed differences in resting brain function network connection,with strong connective regions at the right inferior temporal gyrus,right hippocampus,bilateral thalamus,with weak connective regions at the double medial upper frontal gyrus,lateral dorsal frontal gyurs.Conclusions SED had an effect on the cognitive function of adult patients with epilepsy.The mechanism of cognitive impairment in adult epilepsy with SED may be related to abnormal brain function in cognitive-related areas.

To revise GBZ 188 Technical Specification for Occupational Health Surveillance based on national laws, regulations, standards, specifications and legal documents of occupational disease, and combination with the actual situation in China. The main modifications are as follows: the occupational health surveillance for workers exposed to toluene (xylene may implement by reference), bromopropane, methyl iodide, ethylene oxide, chloroacetic acid, indium and its compounds, coal tar, coal tarasphalt, asphalt, β-naphthylamine, dust of metal and its compounds(tin, iron, antimony, barium and its compounds), hard metal dust, erionite dust, low temperature, laser, tick-borne encephalitis virus, Borrelia burgdorferi, and human immunodeficiency virus, for scraper or grind operators, and underground workers using squatting or kneeling position, crawling position, side-lying position, or shoulder position for a long period of time are included. The emergency health screening for workers exposed to arsenic, fluorine and its inorganic compounds, and acrylamide are included. The occupational medical examination (OME) for workers exposed to amino and nitro compounds of benzene, phosgene, monomethylamine, organic fluorine and dimethyl sulfate has been adjusted and made mandatory, with corresponding assessments required upon leaving the job. The special occupational health surveillance for workers exposed to mycobacterium tuberculosis and hepatitis virus is removed. The OME conclusion of reexamination is removed, and standardize recheck/additional inspection requirements. The optional items in OME performed before, during and after leaving post are removed, but the optional items in emergency medical examination are retained. Additional OME items are added. The Guideline for OME Summary Reports is added as informative appendix, and so on. The revised GBZ 188 Technical Specification for Occupational Health Surveillance is more scientific and practical.