ObjectiveTo prepare high specific monoclonal antibodies(mAbs) against BP26 of Brucella(B.)melitensis.Methods A recombinant plasmid pET-28a-BP26 was constructed and transformed into competent Escherichia coli BL21 (DE3),and then the bacteria were induced by 1 mmol/L isopropylthio-β-D-galactoside (IPTG).After induction,the recombinant BP26 protein (rBP26) was purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PGAE) and nickel ion affinity chromatography(Ni-NTA).Mice were inoculated with rBP26 antigens for three times at 2-week intervals.The first subcutaneous injection contained 100 μg rBP26 with 0.1 ml complete Freund adjuvant.The second subcutaneous injection was 50 μg rBP26 with 0.1 ml incomplete Freund adjuvant.The antibody titers to rBP26 were determined 2 weeks after each reimmunization.Three days before cell fusion,the mice with the highest titer were intraperitoneally injected with 50 μg rBP26 in 0.1 ml PBS.Pre- and post-immunization sera were collected and used as negative or positive controls for screening mAbs.Mice with the highest titer were sacrificed and spleen cells were isolated.The spleen cells of rBP26 immunized mice were fused with SP2/0 myeloma cells in a ratio of 5 ∶ 1 by polyethylene glycol(PEG) 1450.Antibody-producing hybridomas were primarily screened by an indirect enzyme-linked immunosorbnent assay(ELISA) with rBP26.Reactive hybridomas were subcloned for 3 times,then the strains of hybridoma cells secreting antibodies against BP26 were obtained.Supernatant of cloned hybridoma cultures was collected for mAb analyses.These mAbs were named by the hybridoma clone number and tested their reactivity to membrane proteins extracted(NMP) from B.melitensis vaccine strain(M5-90) by Western blotting and Dot-ELISA.mAbs isotyping and kappa(κ) or lambda(λ) light chain was identified by Mouse Monoclonal Antibody Isotyping Kit.Results A total of two mAbs reactive to rBP26 of B.melitensis were selected from antibody screening hybridomas by indirect-ELISA.The two mAbs were named 3C3 and 5A5,and identified as IgG1 (κ) and IgG2(κ),respectively.They could react with NMP from M5-90.Conclusions Results of identification show that two mAbs against rBP26 can be produced.The two mAbs can recognize natural BP26 protein,giving the experimental materials for further research on identification of its epitopes.

Adenocarcinoma ; metabolism ; pathology ; Chromogranin A ; metabolism ; Colonic Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; methods ; Proliferating Cell Nuclear Antigen ; metabolism ; Staining and Labeling ; methods ; Vimentin ; metabolism

Adenosine Triphosphate ; metabolism ; Animals ; Animals, Newborn ; Cantharidin ; pharmacokinetics ; Cell Hypoxia ; drug effects ; Enzyme Inhibitors ; pharmacology ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Flow Cytometry ; Kidney Tubules ; drug effects ; metabolism ; pathology ; Swine

Objective Through the investigation and analysis,Find the influencing factors of nosocomial infection,seek to control the incidence of nosocomial infection methods and countermeasures.Methods Use retrospective study to find 293 infection cases,and 286 normal cases,then process Logistic regression analysis of the second category.Results The influencing factors of nosocomial infection are:the number of days hospitalized,ICU days,age,blood transfusion,the number of days serious ill,the sections change times.Conclusion In order to control the incidence of nosocomial infection,focus on the crowd of high-risk factors,bring in the performance appraisal,establish the HIS to realtime monitor.

Objective To explore the roles of macrophage inflammatory protein-1?(MIP-1?)in hypoxic-ischemic encephalopathy(HIE)of newborn infarnts.Methods Serum samples were obtained in 24,72 h and 7 d after birth respectively from 34 newborn infants with HIE,and 20 newborn infants without HIE as control group.Enzyme-linked immunosorbent assay(ELISA)method was used to determine the serum concentrations of MIP-1?.Results Levels of MIP-1? in newborn infants with HIE [(12.47?2.51)ng/L]were significantly higher than that of newborn infants without HIE [(8.63?2.63)ng/L](P0.05).Conclusions MIP-1? are involved in HIE of neonates,and the more severe damage,the higher levels in serum,which suggests that,as an inflammatory mediator,the MIP-1? may play an important role in involvement of brain hypoxic-ischemic damage.

Objective:To investigate the predictive value of serum soluble urokinase-type plasminogen activator receptor (suPAR) combined with alpha-fetoprotein (AFP) and model for end-stage liver disease (MELD) score in short-term prognosis assessment of patients with chronic hepatitis B (CHB) related acute-on-chronic liver failure (ACLF).Methods:From January 2018 to May 2020, 66 patients with CHB related ACLF from Fuzhou First People′s Hospital were enrolled. After 90 days of follow-up, the patients with CHB related ACLF were divided into death group and survival group according to the outcome. Meanwhile, 30 patients with CHB were enrolled by simple random sampling method. The differences of serum suPAR in patients with CHB related ACLF and patients with CHB were analyzed. The values of suPAR, AFP and MELD score were compared between death group and survival group in patients with CHB related ACLF. The predictive value of suPAR, AFP, MELD score, Child-Turcotte Pugh score (CTP score) and suPAR combined with AFP and MELD score in the short-term prognosis of patients with CHB related ACLF were analyzed by area under the receiver operator characteristic curve (AUROC). Data were analyzed by two independent sample t test or non-parametric test. Results:The serum suPAR level of patients with CHB related ACLF was (9.6±0.8) ln ng/L, which was higher than that of patients with CHB ((8.0±0.3) ln ng/L). The difference was statistically significant ( t=14.533, P<0.01). The suPAR and MELD score of patients with CHB related ACLF in the death group were (9.9±0.7) ln ng/L and 29.6 (7.1) points, respectively, which were higher than those in the survival group ((9.4±0.7) ln ng/L and 21.0 (5.0) points, respectively). The AFP level in the death group was 45.9 (108.1) μg/L, which was lower than that in the survival group (209.3 (187.1) μg/L). There were significant differences in suPAR ( t=2.895, P=0.005), MELD score ( Z=4.708, P<0.01) and AFP ( Z=3.051, P<0.01) between the death group and the survival group. AUROC of suPAR (0.741, P=0.001), AFP (0.724, P=0.002) and MELD score (0.885, P<0.01) had predictive value for death in patients with CHB related ACLF. The sensitivities of suPAR, AFP, MELD score, CTP score and suPAR combined with AFP and MELD score were 84.6%, 73.1%, 88.5%, 96.2% and 84.6%, respectively, and the specificities were 75.0%, 72.5%, 70.0%, 52.5% and 92.5%, respectively. The AUROC of suPAR combined with AFP and MELD score was 0.871 ( P<0.01), which was higher than that of CTP score (0.793, P<0.01). Conclusions:Serum suPAR is increased in patients with CHB related ACLF. SuPAR combined with AFP and MELD score could apply in the prognostic value for patients with CHB related ACLF.

Objective:To investigate the alteration of delayed memory and its relationship with neurogenesis and angiogenesis in vascular dementia (VD) rats after moxibustion therapy. Methods: Two hundred adult male SPF Wistar rats were chosen for the experiment. Thirty-six rats were randomly selected as the sham operation group. Except for rats in the sham operation group (n=36), the others were made into VD models by bilateral common carotid arteries occlusion (BCCAo). After modeling, the 108 survived rats were randomly divided into 3 groups: a model group, a neural stem cells (NSCs) plus endothelial progenitor cells (EPCs) moxibustion group and a NSCs moxibustion group. Co-transplanted implant was transplanted into the rats in the NSCs plus EPCs moxibustion group, and the rats in the NSCs moxibustion group were transplanted by NSCs only. The NSCs plus EPCs moxibustion group and the NSCs moxibustion group received suspended moxibustion therapy at Baihui (GV 20), Dazhui (GV 14) and Shenting (GV 24), (each group was divided into 3 subgroups by the treatment course as 1, 2 and 3 courses). Every group was measured by Morris water maze to evaluate its delayed memory after 3 treatment courses and the rat’s brain was taken out after perfusion of 4% paraformaldehyde one day after 1, 2 and 3 treatment courses, respectively. Marker protein expression was detected by laser confocal microscope to analyze the effect on neurogenesis and angiogenesis. Results: VD rats showed delayed memory in Morris water maze test 3 d after ischemic injury. After 3 courses of moxibustion therapy, VD-induced delayed memory deficits were improved in the NSCs plus EPCs moxibustion group and the NSCs moxibustion group. The expressions of nestin, doublecortin (DCX) and CD34 increased significantly in the two moxibusiton groups after every treatment course (all P＜0.05), which might contribute to the neurogenesis and angiogenesis in hippocampus. In addition, compared with the rats in the NSCs moxibustion group, the expressions of nestin, DCX and CD34 increased significantly in the NSCs plus EPCs moxibustion group (P＜0.05). Conclusion: Moxibustion can reverse VD-induced delayed memory deficits, which may be related to the promotion of neurogenesis and angiogenesis.

OBJECTIVE: To assess the reproducibility of non-verbal facial expressions (smile lips closed, smile lips open, lip purse, cheek puff) in normal persons using dynamic three-dimensional (3D) imaging and provide reference data for future research. METHODS: In this study, 15 adults (7 males and 8 females) without facial asymmetry and facial nerve dysfunction were recruited. Each participant was seated upright in front of the 3D imaging system in natural head position. The whole face could be captured in all six cameras. The dynamic 3D system captured 60 3D images per second. Four facial expressions were included: smile lips closed, smile lips open, lip purse, and cheek puff. Before starting, we instructed the subjects to make facial expressions to develop muscle memory. During recording, each facial expression took about 3 to 4 seconds. At least 1 week later, the procedure was repeated. The rest position (T0) was considered as the base frame. The first quartile of expressions (T1), just after reaching the maximum state of expressions (T2), just before the end of maximum state of expressions (T3), the third quartile of expressions (T4), and the end of motion (T5) were selected as key frames. Using the stable part of face such as forehead, each key frame (T1-T5) of the different expressions was aligned on the corresponding frame at rest (T0). The root mean square (RMS) between each key frame and its corresponding frame at rest were calculated. The Wilcoxon signed ranks test was applied to assess statistical differences between the corresponding frames of the different facial expressions. RESULTS: Facial expressions like smile lips closed, smile lips open, and cheek puff were reproducible. Lip purse was not reproducible. The statistically significant differences were found on the T2 frame of the repeated lip purse movement. CONCLUSION The dynamic 3D imaging can be used to evaluate the reproducibility of facial expressions. Compared with the qualitative analysis and two-dimensions analysis, dynamic 3D images can be able to more truly represent the facial expressions which make the research more reliable.
Adult ; Face/diagnostic imaging* ; Facial Expression ; Female ; Humans ; Imaging, Three-Dimensional ; Lip/diagnostic imaging* ; Male ; Photogrammetry ; Reproducibility of Results ; Smiling

Animals ; Aorta ; pathology ; Atherosclerosis ; blood ; etiology ; metabolism ; Chemokine CCL2 ; metabolism ; Cholesterol ; blood ; Cholesterol, Dietary ; administration & dosage ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Dehydroepiandrosterone ; pharmacology ; Diet, Atherogenic ; Immunohistochemistry ; Rabbits ; Random Allocation ; Triglycerides ; blood ; Vascular Cell Adhesion Molecule-1 ; metabolism

Objective To evaluate the indications and the surgical technique of endoscopic transsphenoidal approach in the surgical treatment of pituitary adenomas. Methods We retrospectively analyzed the surgical effectiveness and complications of transsphenoidal approach under endoscope in treating 39 patients suffering from pituitary adenomas, including 37 with functional adenomas and 2 with non-functional adenomas. Results In all the patients with functional adenomas,tumors were totally resected. The 2 patients with non-functional adenoma invaded the seller obtained partial resection, and thus, postoperative radiotherapy was performed and preoperative symptoms were improved. All patients had no serious complications. All cases were followed up for 6-36 months: 1 had temporally visual loss; 11 appeared transient polyuria or diabetes insipidus; 2 developed cerebrospinal rhinorrhea, and they were cured after the treatment; headache was disappeared or relieved in 11, and sexual function got improvement in 7; the high preoperative PRL level was decreased to normal value in 37 patients after the surgery. Conclusion Endoscopic transsphenoidal technique, a safe, minimally invasive and efficient procedure for removing pituitary adenomas, offers simple and rapid access to the sella turcica and also provide direct visualization, thus improving the surgeon's ability to resect the tumors.