Objective To investigate the effects of human Rrain-derived neurotrophic factor-gamma fetopro-tein (hBDNF-GFP) gene-transfected neural stem cell (NSC) transplantation on BDNF expressions in the retina of rots after optic nerve (ON) crush injury. Method ①Seventy-eight Sprague-Dawley (SD) rats were randomly as-signed into a control group (n = 6) and ON crush group (n = 72). In the ON crush group, the right ON was crushed while the left NO was exposed as sham injury. Rats in the ON crush group were divided into three sub-groups: PBS group (intravitreons injection of 0.01 mol/L phosphate buffered solution); GFP group (intravitreous transplantation of GFP gene-transfected NSCs); and hBDNF-GFP group (intravitreous transplantation of hBDNF-GFP gene-transfected NSCs). Rats were sacrificed 3, 7, 14 and 28 days after transplantation, and BDNF expres-sions in retinal homogenates was detected by using enzyme-linked immunosorbent assay (ELISA). ②The hBDNF-GFP-NSCs were transplanted intravitreous into six rats after ON crush injury. Following this, two rats were sacri-riced 2, 4 and 8 weeks after transplantation. The survival and location of NSCs in host retina were observed by frozen section analysis. ③Adult SD rats were randomly divided into four groups: control group (n = 5); NSC group (NSC transplantation, n = 10); GFP-NSC group (GFP-NSC transplantation, n = 10); and hBDNF-GFP-NSC group (hBDNF-GFP-NSC transplantation, n = 10). Four and eight weeks after transplantation, five rats from every group were sacrificed. Western blot analysis was used to determine retinal BDNF expression. Results ① There was no significant difference in BDNF expression between the control group and sham-injury groups (P >0.05). Three days after NSC transplantation, BDNF expression increased significantly in the three injured sub-groups compared with the sham-injury group, (P < 0.05), whereas no significant inter-group differences in BDNF expressions among three injured sub-groups were observed (P > 0.05). Seven days after transplantation, there was a significant difference in BDNF expression between the GFP-NSC group and the sham-injury groups (P <0.05), whereas there were no significant differences in BDNF expressions among the PBS, hBDNF-GFP-NSC and sham-injury groups (P > 0.05). Fourteen and 28 days after transplantation, BDNF expressions decreased in the PBS group and the GFP-NSC groups, while BDNF expressions in the hBDNF-GFP-NSC group increased significant-ly compared with the other three groups (P < 0.05); ②Frozen section showed that transplanted hBDNF-GFP-NSCs could survive and gradually extended to all layers of the host retina. ③Westem blot revealed there were no differences in BDNF expressions between 4-week and 8-week intervals in the hBDNF-GFP-NSC group. Compared with other three groups, BDNF expressions in the retina increased significantly after hBDNF-GFP-NSC transplanta-tion. Conclusions The hBDNF-GFP gene-trausfected NSCs can survive in the host retina and BDNF expressions are stable at a high level.

BACKGROUND: Myelin sheath related inhibitors have been found to have great impact on microenvironment of axon regeneration. Traditional Chinese medicine is gradual y becoming a research hotspot on improving microenvironment of nerve regeneration with its advantage on multiple factors and targets.
OBJECTIVE: To clarify the effect of Jisuikang on Nogo-66 receptor NgR expression after spinal cord injury.
METHODS: 144 rats were randomly divided into six groups: sham surgery group, model group, prednisone group, high-, moderate- and low-dose Jisuikang groups (n=24). Animal models of spinal cord injury were established by the modified Allen’s method in the later five groups. Rats in the prednisone group were daily given 0.06 g/kg prednisone acetate by lavage, once a day. Rats in the high-, moderate- and low-dose Jisuikang groups were daily intragastrically given 12.5, 25 and 50 g/kg Jisuikang, once a day. Rats in the sham surgery and model groups were intragastrically daily given 20 mL of saline, once a day. Rats in each group were administered drugs until death.
RESULTS AND CONCLUSION: Compared with the model group, NgR protein and mRNA expression levels were significantly lower in the prednisone, moderate-and low-dose Jisuikang groups. These data suggested that Jisuikang can improve the recovery of neurological function after spinal cord injury and effectively inhibit NgR protein expression at the site of injury so as to suppress the microenvironment factors harmful to nerve regeneration and further improve the microenvironment of nerve regeneration. Subject headings: Drugs, Chinese Herbal; Spinal Cord Injuries; Axons; Tissue Engineering

OBJECTIVE:To provide reference for rational use of human serum albumin for surgical inpatients. METHODS:The utilization of human serum albumin for surgical inpatients in a hospital during Jan.-Mar. 2014 was analyzed and evaluated by UHC Guidelines for the Use of Album,Nonprotein Colloid,and Crystalloid Solutions(2010 edition)and European Immune Globu-lin and Albumin Use Recommendation. RESULTS:Among the 556 patients,totally 895 human serum albumin application were con-ducted,mainly involving development of gastrointestinal surgery(29.7%),hepatobiliary surgery(25.9%)and cardiothoracic sur-gery(13.1%). The main reasons were correcting hypoalbuminemia(62.9%),followed by albumin supplemented during major sur-gery(7.9%)and alleviating ascites in patients with cirrhosis(4.4%);only 95 applications(10.6%)were considered appropriate. The most prevalent inappropriate reason was for correcting hypoalbuminemia. CONCLUSIONS:Human serum albumin in the surgi-cal inpatients in the hospital shows a large amount,and low consistent rate between indications and guidelines. The rational stan-dardized utilization of human serum albumin should be strengthened.

OBJECTIVETo observe the protection effect of Ligustrazine Hydrochloride (LH) on coagulation reaction and inflammation reaction in single valve replacement patients with rheumatic heart disease undergoing cardiopulmonary bypass (CPB).

METHODSTotally 40 patients undergoing single valve replacement were recruited in the study and randomly assigned to the two groups, the treatment group and the control group, 20 in each group. In treatment group LH (3 mg/kg) was intravenously infused from the jugular vein. LH (3 mg/kg) was also added in the CPB priming. In the control group LH was replaced by equal amount of normal saline. Endothelial micro-particles (EMP) count was detected before CPB, 30 min after CPB, 1 h and 24 h after CPB finished. The coagulation reaction time (R), coagulation time (K), clotting formation velocity (alpha angle), maximum amplitude (MA), coagulation index (CI), platelet (PLT), hypersensitive C reactive protein (hs-CRP), IL-6, and IL-10 were detected before CPB, 1 h and 24 h after CPB finished.

RESULTSThere was no statistical difference in aorta arresting time, period of CPB, post-operative drainage volume, plasma transfusion volume, post-operative respirator assistant time, and hospitalization time between the two groups (P >0.05). Compared with pre-CPB in the same group, the count of EMP was much higher at 30 min after CPB and 1 h after CPB finished (P < 0.01). R and K, hs-CRP, IL-6, and IL-10 increased at 1 h and 24 h after CPB finished (P <0.01,P < 0.05). The alpha angle,.MA, CI, and PLT decreased 1 h after CPB finished (P <0.01). The a angle increased, while CI and PLT decreased 24 h after CPB finished (P <0.05). Compared with the control group in the same period, the count of EMP was lower in the treatment group 30 min after CPB and 1 h after CPB finished (P <0. 05, P <0. 01). R and K values obviously decreased in treatment group 1 hour after CPB finished (P <0. 05), while a angle, MA, CI, and PLT increased (P <0. 05, P <0. 01). hs-CRP and IL-6 decreased in the treatment group 1 h and 24 h after CPB finished (P <0.05), while IL-10 increased (P <0.05). The count of PLT increased 24 h after CPB finished in the treatment group (P <0. 05).

CONCLUSIONLH had certain protection effect on the vascular endothelium undergoing CPB, and lower excessive activation of coagulation reaction and inflammation reaction in patients undergoing CPB.

Blood Coagulation ; drug effects ; C-Reactive Protein ; metabolism ; Cardiopulmonary Bypass ; methods ; Humans ; Inflammation ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Pyrazines ; pharmacology ; therapeutic use ; Rheumatic Heart Disease ; drug therapy

Adult ; Antidepressive Agents ; therapeutic use ; Depression ; complications ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Dyspepsia ; drug therapy ; psychology ; Female ; Gastric Emptying ; drug effects ; Gastrointestinal Agents ; therapeutic use ; Gastrointestinal Motility ; drug effects ; Humans ; Male ; Middle Aged ; Phytotherapy ; Powders

To find a rapid and accurate genotyping method for specific non-syndromic hearing loss (NSHL)-causing gene mutations for disease diagnosis in different ethnic populations.Methods We performed a novel multiplex primer extension (PE) reaction in combination with denaturing high-performance liquid chromatography (DHPLC) to simultaneously detect and genotype the 6 most common mutations in 180 patients with NSHL (GJB2-235delC,GJB2-299delAT,PDS-A2168G,PDS IVS7-2A ＞ G,mtDNA-A1555G,and mtDNA-C1494T) in Chinese population.This method involved the amplification of the target sequence,followed by a purification step,a multiplex PE reaction,and DHPLC analysis performed on the Transgenomic Wave DNA fragment analysis system under fully-denaturing conditions.Results In a blind analysis,this technique successfully and accurately genotyped 100％ of the samples simultaneously characterized by direct sequencing.Conclusion Combination of PE and DHPLC is simple,rapid,accurate,and cost-effective for genotyping common disease-causing mutations,including substitutions,insertions,and deletions in NSHL,and may be successfully used in other genetic diseases.

Objective To study the characteristics of the transcranial electric stimulation(TES)-elicited motor evoked potentials(MEP)recorded from different points of the diaphragm,and to determine the optimal record- ing site in the diaphragm.Methods One EEG electrode was placed subcutaneously in the midline of the skull as stimulation electrode and another EEG electrode was inserted into hard palate submucously as cathode in 30 male rats.And single square wave electrical pulse was used for stimulation.The stimulation intensity was 15 mA,the du- ration of the pulse was 0.2 ms,and the stimulating interval was 200 ms.The concentric needle electrode were insert- ed into the following eight sites:the medial,intermediate,lateral crura of the diaphragm,the junction of the posterior axilla line and the inferior border of the eleventh rib,the junction of the anterior axilla line and the inferior border of the ninth rib,the junction of the intermediate line of the clavicle and the inferior border of the eighth rib,the junction of the para-sternal line and the inferior border of the seventh rib,the pars sternalis.The MEP was recorded from each point,respectively.The optimal sites of the diaphragm MEP were found and recorded.Results The MEPs were re- corded from each of the 8 sites of the diaphragm in all the rats after TES.There was no statistically significant differ- ence among the latencies of every site.But the amplitude varied from 6.08?0.46 mV in maximum along the inferior border of the ninth rib at the anterior line of the axilla to the minimum of O.91?0.03 mV at the pars sternalis. Conclusion It is safer to insert the needle along the inferior border of the ninth rib at the anterior line of the axilla, because there was certain distant from the folding part of the pleura and the amplitude of the recorded wave at this point is maximal.

Coprinus comatus cultivated on cotton seed hull medium decomposed lignocellulose straggly and was high of absolute biological efficiency. Lignocellulose is the main carbon source for the fruiting stage of the fungus. There existed the positive correlation between the degradation rates of the cellulose and hemicellulose in the medium and the activities of extracellular CMCase (carboxymenthelcel lulase), FPase (filter paper cellulase) and HCase (hemicellu-lase), there also existed the positive correlation between the degradation rate of the lignin in the medium and the activity of extracellular laccase, but no correlation between the degradatio rate of the lignin in the medium and the activity of peroxidase. The activity of extracellular amylase was comparatively high at mycelial growth stage, and the protease activity peek was at teh time when the fruitbody matured.

Objective To prepare specific polyclonal antibodies to outer membrane protein (Opr) D_2 of Pseudomonas aeruginosa (PA),and explore the relationship between loss of OprD_2 and imipenem resistance.Methods The genomic DNA of PA was ex- tracted with phenol:chloroform.OprD_2 coding gene was amplified by PCR and prokaryotic expression vector pRSET-OprD_2 was constructed.OprD_2 protein was expressed by IPTG induction in E.coli BL21(DE3),and purified with SDS-PAGE.The new protein band was recovered and used as antigens to subcutaneously immunize two New Zealand rabbits to prepare poly- clonal antibody.The specificity of the antibody was determined by Western blot.The expression of OprD_2 in 32 clinical isolates of PA was detected with the prepared polyclonal antibody by Western blot.Results The vector pRSET-OprD_2 has been success- fully expressed in E.coli BL21 (DE3).The polyclonal anti-OprD_2 antibody with high specificity has been successfully pre- pared.Present results show that of the 27 imipenem-resistant PA clinical isolates,OprD2 protein was low-expressed in 5 iso- lates (18.5%) and normally expressed in 2 isolates (7.4%) but not expressed in 20 isolates (74.1%).Conclusions The loss or low-expression of OprD_2 is one of the essential mechanisms accounting for imipenem resistance in clinical isolates of PA.

Objective: To investigate the protective effect of rhubarb (大黄) combined with scopolamine on gastric and gut mucosal barrier in patients with severe trauma. Methods: Forty patients with severe trauma, injury severity score (ISS)≥25, were randomly divided into two groups (each n=20), namely common therapy group (CM group) and rhubarb (infused into stomach) combined with scopolamine group (RS group). In RS group, rhubarb combined with scopolamine were given just after the first time measuring gastrointramucosal pH (pHi), the other treatment was the same as CM group.The mean arterial pressure (MAP) and heart rate (HR) were recorded, and pHi, lactic acid, interleukin-6 (IL-6) and tumor necrosis factor-? (TNF-?) were measured on admission and at 24, 48 and 72 hours after admission. Results: The MAP, HR and pHi were uptrend in both groups. There were significant increase in MAP and pHi of RS group at 72 hours compared with those of CM group (both P