This paper described the results of the optimal method for preparation and extraction of lectin from Phaeoporus obliquus(Pers ex Fr).(J.Schroet).We determined the optimal buffer solution using the ag-glutination test with 2% hare blood cells.The optimal extraction process included the following parameters that were conformed by optimization analysis: material-water ratio,extraction time,concentration of sodium chloride and the pH value.The results are that the agglutination indexes of POL using TBS and PBS extrac-tion buffer were 64 and 16,respectively.The optimal extraction process is that,the material-water ratio was 1:50,the extraction time was 20 h,while the concentration of sodium chloride was 0 mol/L and pH was 8.0.The agglutinability of POL was 256 examined by this method.The optimized extraction process is stable and practicable,which may supply some basis for the application of the lectin from Phaeoporus obliquus in immunoregulation.

Objective To investigate the effect of brain-derived neurotrophic factor (BDNF) gene modified neural stem cells (NSCs)transplantation on cerebral ischemic injury in rats.Methods NSCs from newborn rat hippocampus were isolated,cultured in a medium containing fibroblast factor (FGF) in vitro. Their proliferation and differentiation were detected by immunohistochemistry.Virus vectors pLXSN-BDNF were built and BDNF were transfected into NSCs.Biological activity were detected to obtained engineering stem cells of BDNF protein with secretary activity.Middle cerebral artery occlusion (MCAO) models were made and transplanted with NSCs-BDNF by stereotaxic technique.The effect of transplantation on MCAO models was evaluated histologically and behaviorally.Results Statistical analysis showed that the behavioral performance of the animals improved after transplantation (Longa mark being 1.343?0.293 four weeks after stem cell transplantation).The number of hippocampal dentate gyrus neurons increased to 87.5%?6.6% , four weeks after stem cell transplantation on Nissle stained hippocampal sections,which was significantly different from that of controls.Positively BrdU stained neural stem cells revealed by immunohistochemistry in the cultured cells and in the transplanted brain sections,indicated that the engineering cells transplanted had survived in the host brain and began to proliferate.Conclusion Transplantation of BDNF genetically modified NSCs can effectively promote the recovery from cerebral ischemic injury.

Background The popularization and promotion of gene diagnosis technology makes it possible to detect deafness genes for children with congenital hearing impairment,and the proportion of gap junction protein beta 2 (GJB2) gene mutations in cochlear implant patients is 26.5％ We did follow-up evaluation on auditory rehabilitation effect for all 31 deaf children with GJB2 gene mutation after cochlear implantation to provide a reference for such patients.Methods Application of “the genetic deafness gene chip detection kit” and “gene complete sequence analysis” were applied to conduct detection on common genetic deafness gene mutation hotspots of the hearing impaired children with cochlear implantation.To conduct auditory rehabilitation effect evaluation on all 31 cases of patients with GJB2 genetic deafness after 3,6 and 12 months of the operation respectively.The single factor repeated measure analysis of variance (ANOVA) was applied to analysis whether there were significant difference among the results of initial consonant of a Chinese syllable recognition at 3 different stages after the operation,the results of vowel of a Chinese syllable recognition at 3 different stages after the operation,and the results of two-syllable recognition at 3 different stages after the operation.Results The 235delC is the high-incidence mutational site in 31 cases of patients with GJB2 genetic deafness,and the total detection rate is up to 90.3％ (28/31).There were significant differences in the initial consonant and the vowel of a Chinese syllable recognition rate,and the two-syllable recognition rates at 3,6,and 12 months after the operation (P＜0.01).Conclusion Cochlear implantation is a safe and effective measure for auditory reconstruction,enabling patients with GJB2 hereditary severe sensorineural deafness to achieve auditory speech recognition effectively.

Background: Post-operative pain is unpleasant for patients and may worsen surgical recovery.Peri-operative multimodal analgesia has been used for many years; however,its efficacy still needs improvement.In the present study,a thorough peri-operative pain counseling and stratified management program based on risk assessment was implemented,with the goal of improving post-operative analgesia and patient satisfaction.Methods: This prospective,controlled,pilot study included 361 patients who underwent elective surgery.Of these 361 patients,187 received peri-operative pain risk assessment and stratified analgesia and counseling(stratified analgesia group),while 174 received conventional multimodal analgesia(conventional group).The two groups were compared regarding the post-operative pain intensity,rescue analgesia administration,post-operative quality of recovery as assessed via the quality of recovery 40 questionnaire,total dosage of peri-operative opioids,analgesic satisfaction,and analgesic costs.Results: Compared with the conventional group,the stratified analgesia group reported decreased pain intensity during motion at 24 h post-operatively and required lower dosages of rescue analgesia(P = 0.03).The total quality of recovery 40 questionnaire score and the scores for physical wellbeing and pain were significantly better in the stratified analgesia group than the conventional group(P = 0.04); the stratified analgesia group also reported better scores for analgesic satisfaction(P = 0.03)and received lower dosages of opioids(P = 0.03).Analgesic costs were lower in the stratified analgesia group than the conventional group; the cost-effective ratio was 109 in the conventional group and 62 in the stratified analgesia group.Conclusions: The analgesic efficacy was improved by the implementation of stratified analgesia based on surgical pain risk assessment and counseling.This stratified analgesia protocol increased the patients' analgesic satisfaction and improved the quality of recovery without increasing healthcare costs.The present findings may help improve the efficacy of peri-operative multimodal analgesia in clinical practice.

BACKGROUND AND OBJECTIVEPrevious studies have shown that Bmi-1 is overexpressed in a variety of tumors, suggesting that Bmi-1 plays an important role in tumorigenesis. In this study, we investigated the effect of Bim-1 siRNA on cell proliferation, cell cycle, cell apoptosis and migration of human esophageal carcinoma EC9706 cells, and explored its potential mechanisms.

METHODSBmi-1 small interfering RNA (siRNA) was transferred into EC9706 cells. Then, cell proliferation was measured using cell counting kit-8 (CCK-8), cell cycle and cell apoptosis were analyzed by flow cytometry, cell migration ability was detected using Boyden chamber assay, and the mRNA and protein expression levels of Bmi-1, p16, Bcl-2, Bax, and MMP-2 were determined using real-time polymerase chain reaction (PCR) and Western blot analysis, respectively.

RESULTSBmi-1 siRNA treatment significantly inhibited the expression of Bmi-1 at both mRNA and protein levels in EC9706 cells. Cell proliferation rate decreased dramatically in the Bmi-1 siRNA treated group than in the untreated group and in the scrambled siRNA treated group (both P < 0.001). In Bmi-1 treated group, the percentage of cells at G(0)/G(1) stage was 71.93%, which was higher than that in the untreated group (47.36%) or scramble siRNA treated group (48.47%) (both P < 0.001). Early cell apoptosis rate also increased significantly in the Bmi-1 siRNA treated group (both 17.32%) than in the untreated group (2.61%) and in the scramble siRNA treated group (2.73%) (both P < 0.001). Further experiment suggested that downregulation of Bmi-1 led to less cell migration. In EC9706 cells transfected by Bmi-1 siRNA, the expression levels of p16 and Bax increased, while the expression level of Bcl-2 decreased.

CONCLUSIONSBmi-1 downregulation in esophageal carcinoma cells inhibits cell proliferation, cell cycle, and cell migration, while increases cell apoptosis. These results suggest that Bmi-1 is a potential molecular target of treating esophageal cancer.

Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Down-Regulation ; Esophageal Neoplasms ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Nuclear Proteins ; genetics ; metabolism ; physiology ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; metabolism ; physiology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Repressor Proteins ; genetics ; metabolism ; physiology ; Transfection ; bcl-2-Associated X Protein ; metabolism

Objective To analyzed the computed tomography angiography (CTA) features of the coronary artery fistulas. Methods Sixty-six coronary artery fistulas were diagnosed out of 12 717 patients underwent the coronary artery multiple detector CTA examination. The origin and drainage site of the coronary artery fistulas and the plaque and stenosis of the coronary artery were observed by post-processing analysis on various images. Coronary artery angiography was performed in 14 out of 66 coronary artery fistulas patients. Results Coronary artery fistulas arose from bilateral coronary artery system in 21 cases, from left coronary artery in 26 cases and from right coronary artery in 19 cases. The majority of coronary artery fistulas entered into pulmonary artery (41 cases). The rest drainage sites included left atrium (10 cases), right atrium (8 cases),left ventricle (4 cases), coronary sinus (2 cases) and right ventricle (1 case). The findings of CTA and coronary artery angiography were consistent in 14 patients with DSA examination. Coronary artery plagues were evidenced in 31 cases and stenosis was greater than 50% in 7 coronary artery fistulas patients. Conclusions Multiple coronary artery fistulas are not rare, and pulmonary artery is the most frequent drainage site. When suspecting the coronary artery fistulas, coronary artery CTA can be the first choice of diagnose. CTA can supply adequate information for therapy.

Adult ; Biomarkers ; blood ; Case-Control Studies ; Female ; Granzymes ; blood ; Hepatitis B ; blood ; immunology ; Humans ; Male ; Middle Aged ; Perforin ; blood ; T-Lymphocytes, Cytotoxic ; metabolism

Objective Clinical data of 19 Chinese patients with 21 hydroxylase deficiency (21OHD) were analyzed to improve the diagnosis and treatment level. Methods Clinical features and laboratory data were collected from 19 patients with 21OHD before and after treatment. Results In male patients, the average age of early appearance of secondary sexual character was (9.3?2.8)yrs, and excess androgen resulted in phallic enlargement. Primary amenorrhea was the most common complaint in female(87.5%), and the signs included a varying degree of labioscrotal fusion and clitoral enlargement. The average level of 17-hydroxy progesterone(17OHP) was (63.42?35.07) ?g/L, and adrenocorticotrophic hormone(ACTH), dehydroepiandrosterone(sodium) sulfate(DHEAS) and testosterone(T) were obviously elevated. CT scan showed bilateral adrenal hyperplasia. The level of 17OHP was significantly decreased after treatment[(63.42?35.07) ?g/L vs (3.15?2.71) ?g/L](P

Objective To detect the feasibility of magnetically labeled swine bone marrow mesenehymal stem cells(MSCs)with SHU 555A combined with poly-L-arginine(PLL),under MR imaging in vitro and in vivo.Methods Swine mesenehymal stem cells were isolated and culture-expanded 3 passages in vitro,then magnetically labeled by incubation with SHU 555A(25?g Fe/ml,Resovist,Schering)for 24 hours with 750 ng/mL poly-L-lysine(PLL;average MW_275 kDa)added 1 hour before incubation.Cellular iron incorporation and detention at 0 d,4 d,8 d,12 d,16 d,20 d after labeling was qualitatively assessed using Prussian blue and quantified at atomic absorption spectrometry.Cell viability was assessed by trypan-blue exclusion test.Cell suspensions underwent MR imaging with T_1-and T_2-weighted spin-echo and fast field-echo sequences on a clinical 1.5 T MR system.At last,1?10~6 SHU 555A labeled and unlabeled MSCs were transextracardially implanted into the infracted and normal myocardium approximately 2 week following the ligation of left anterior descending coronary artery in 1 swine respectively,and finally performed 1.5-T MRI within 1 week after infarction.Results①Intracytoplasmic particles stained with Prussian blue stain were detected for all cells with mean cellular iron content of(13.13?2.30)pg per cell.With division of stem cells, the stained particles decreased gradually with iron content(0.68?0.20)pg per cell.at 16 days after labeling, approximately to the prelabeled baseline values.(0.21?0.06)pg per cell(P＞0.05).The viability of the labeled cells at various time points were not significantly different with that of nonlabeled cells(P＞0.05).②MR images showed signal intensity changed most obviouly in T2*WI in vitro.The percentage change of signal intensity increased with increasing cell numbers,and decreased with the time.As few as 5?10~4-1?10~5 cells could be detected by using this approach.③Two injected sites containing MR-MSCs were detected in vivo,presentingas low signal intensity areas with the T_2*WI scanning sequence.Conclusion Swine bone marrow MSCs can be labeled with SHU555A-PLL and depicted with a standard 1.5-T MR imager in vitro and in vivo.(J lntervent Radiol,2007,16:115-121)

Objective The present study was conducted to investigate the infection of Lyme disease, Spotted fever, Ehrlichiosis (anaplasmosisin) in wild animals and ticks in the mountain areas of Zhejiang province. Methods Nested polymerase chain reaction was used to amplify specific DNA sequences of Lyme spirochetes, Spotted fever group rickettsiae, Ehrlichia (anaplasma) from samples of mice and ticks. Results 14 positive samples were identified from 121 mice and 105 groups of ticks. Among mice samples, one positive 5S-23S rDNA intergenic spacer of Borreia burgdorferi and two 5' fragments of Ehrlichia (anaplasma) 16S rDNA were obtained. 11 positive results were detected from tick samples including three 5S-23S rDNA intergenic spacer regions of Borreia burgdorferi and eight 5' fragments of Spotted fever group rickettsiae outer member protein A gene. One group of adult ticks, Haemaphysalis longicornis, which had been collected from eastern mountain area were detected to have co-infected with Lyme spirochetes and Spotted fever group rickettsiae. The positive sequences of 5S-23S rDNA intergenic spacer and ompA gene were tested and analyzed as Lyme spirochetes while rickettsia which was closely related to Borrelia valaisiana and R. massilliae. Conclusion This was the first report about co-infection of Lyme spirochetes and Spotted fever group rickettsiae found in the same group of adult Haemaphysalis longicornis. It is very important to strengthen the surveillance program on tick-borne infectious disease and their pathogenic in vectors, wild animals and targeted high risk groups and to differentiate the clinical manifestation and diagnosis to extend the knowledge of tick-borne infectious diseases in Zhejiang.