Human herpesvirus 7 (HHV-7) infection is dependent on the functions of structural glycoproteins at multiple stages of the viral life cycle. These proteins mediate the initial attachment and fusion events that occur between the viral envelope and a host cell membrane, as well as cell to cell spread of the virus. To characterize the HHV-7 glycoproteins that can mediate cell fusion, a cell-based fusion assay was used. 293T cells expressing the HHV-7 glycoproteins of interest along with a luciferase reporter gene under the control of the T7 promoter were cocultivated with SupT1 cells transfected with T7 RNA polymerase. HHV-7 glycoproteins gB, gH, gL and gO can mediate the fusion of 293T cells with SupT1 cells, and the fusion can be inhibited by anti-CD4 mAbs. Thus, the coexpression of HHV-7 gB, gO, gH and gL is sufficient and necessary for HHV-7 induced membrane fusion, and one of these glycoproteins or protein complex formed by these glycoproteins might be the ligand(s) of CD4 molecule.

OBJECTIVETo establish and evaluate a BA-ELISA method for the quantitative detection of cystic fibrosis transmembrane conductance regulator (CFTR) protein.

METHODSWe deliberately selected three tables of CFTR and made the synthetic peptide be expressed in E. coli, then used the antigen to immunize rabbits to obtain the anti-CFTR polyclonal serum. After that, 96 well plates were coated with the purified antibody against CFTR. The antigen CFTR which was extracted from human sperm was detected by anti-CFTR antibody labeled with biotin, horseradish peroxidase conjugated avidin, and the substrate. The concentrations of two kinds of antibodies and the experiment parameters were optimized. Thereby, the double antibody sandwich BA-ELISA method for the quantitative detection of CFTR protein was established. Furthermore, the reproducibility, specificity and so on were evaluated by clinical specimens of sperm.

RESULTSThe optimal concentration of coated anti-CFTR IgG was 4 µg/ml, while the biotin labeled anti-CFTR IgG was 10 µg/ml; the optimal blocking buffer was 1% BSA-PBST, the optimal time of the reaction between antigen and antibody was 60 min, the optimal chromogenic time was 15 min, the intra-assay and inter-assay coefficient were 2.16%-9.23% and 2.29%-11.71% respectively; The lowest detectable limit was 0.15 ng/ml; the standard curve had a good linear correlation of R2 = 0.962.

CONCLUSIONThe BA-ELISA method for the quantitative detection of CTFR protein is successfully established, and it is demonstrated that the method has strong specificity, high sensitivity and good reproducibility. It provides the basis and evidence of the further application of the method.

Animals ; Antibodies ; Cystic Fibrosis Transmembrane Conductance Regulator ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; Humans ; Peptides ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity

Objective:To investigate the effect of estrogen on the expression of P2X7 receptor ( P2X7R) in the cerebral cortex and neuroinflammation after subarachnoid hemorrhage ( SAH) in rats.Methods:The rat model of SAH was induced by modified mono-filament puncture method.Sixty male SD rats were divided randomly into three groups:sham group;SAH group and estrogen-treatment group.The local cerebral blood flow was detected with laser doppler blood flow meter.The content of tumor necrosis factor alpha ( TNF-α) and interleukin-6 (IL-6) were measured by ELISA analysis.The expression of P2X7R in the cerebral cortex was tested by immuno-histochemical and Western blot methods.Results:Compared with that in Sham group,cerebral blood flow was significantly decreased after SAH (P<0.05),the content of TNF-αand IL-6 in the cerebral cortex were significantly up-regulated at each time point after SAH (P<0.05),peaked at 24 h,and the expression of P2X7R significantly increased at 6 h,12 h and 24 h after SAH (P<0.05),peaked at 12 h.Compared with that in SAH group,cerebral blood flow was significantly increased in estrogen-treatment group (P<0.05),the levels of TNF-α,IL-6 and P2X7R were down-regulated in estrogen-treatment group ( P<0.05).Conclusion: Estrogen could attenuate neuroinflammation in the cerebral cortex after subarachnoid hemorrhage in rats, which may be associated with the down-regulation in P2X7R proteins.

Objective To assess the value of hemofiltration in the prevention of contrast induced nephropathy(CIN) post percutaneous coronary intervention(PCI) in severe chronic kidney disease stages(CKD) patients. Methods We evaluated 30 CKD patients who underwent PCI followed by bedside hemofiltration. We measured serum creatinine levels before PCI, 24 hours, 72 hours, and one week after PCI, and calculated creatinine clearance(CrCl) according to Cockcroft and Gault equation.We observed the incidence of CIN, and the short term clinical efficacy of hemofiltralion. Results The average age of the 30 patients was (72.87 ± 8.71) years old, with 21 (70%) male patients. The stages of CKD among the patients included CKD 3 (3 patients, 10%), CKD 4 (20 patients, 66.7%) and CKD 5 (7 patients, 23.3%). The average duration of hemofiltration was (7.5±4.1) hours. Serum creatinine before PCI, 24 hours, 72 hours and 1 week after PCI was (498.7±143.7)μmol/L, (353.2±128.0)μmol/L, (450.0±132.2)μmol/L, (488.0±145.7)μmol/L respectively,and CrCl was (20.3±10.2) ml/min, (36.5±14.3) ml/min, (28.3±10.4) ml/min, (21.0±10.3) ml/min respectively. There was no CIN. Mean follow-up was (3.1 ± 2.6) months. There was no new cardiovascular events, and no new patients need to rely on long term hemodialysis. Conclusions For patients with severe renal insufficiency, hemofiltration may reduce the incidence of CIN, It is an alternative preventive measures to prevent CIN.

A diving decompression procedure is a specific rule that divers should follow when they ascend and get out of water. It comes from the decompression theory and algorithm and is designed for the prevention of decompression sickness. With the ， ， development of diving technology and diving medicine the decompression procedures are constantly innovated and the new ， decompression procedure can be used in diving practice after safety verification. In principle the safety verification of ， decompression procedures should be conducted on animal experiments before human experiments and the risks of ， decompression sickness and oxygen toxicity should be systematically assessed. However the assessment methods used in ， ， ， different studies differ greatly thus it is urgent to establish a standard and universal verification system. Traditionally the risk ， ， assessment of decompression sickness and oxygen toxicity is mainly carried out by observing the incidence detecting bubbles ， theoretical calculation and lung functional test. Furthermore biochemical indicators are increasingly becoming important ， ， supplements. Due to the special underwater environment the diving operation is prone to accidents. Therefore in addition to ， verifying the safety of the new decompression procedure exploring its safety decompression limit is of great significance for the formulation of emergency decompression procedures in emergency situations. The specific approach is to shorten the decompression time and assess the safety until the critical time for detecting bubbles without the occurrence of decompression ， ， sickness is found. Future studies should continue to optimize safety assessment methods explore sensitive biochemical markers ， clarify species associations and improve verification efficiency and reliability of results.

OBJECTIVE: To analyze the causes of the esophagogastric junction outlet obstruction (EGJOO) patients, to discuss the differences of the clinical manifestation and esophageal motility characteristics between the anatomic EGJOO (A-EGJOO) and functional EGJOO (F-EGJOO) subgroups, and to search the diagnostic values of the specific metrics for differentiating the subgroups of EGJOO patients. METHODS: For the current retrospective study, all the patients who underwent the esophageal high resonance manometry test were retrospectively analyzed from Jan 2012 to Oct 2018 in Peking University Third Hospital. The EGJOO patients were enrolled in the following research. The clinical characteristics, such as symptoms and causes of the patients were studied. Then the patients were divided into two subgroups as A-EGJOO subgroup and F-EGJOO subgroup. The clinical symptoms and the main manometry metrics were compared between these two subgroups. The significant different metrics between the two groups were selected to draw receiver operating characteristic (ROC) curves and the diagnostic values were analyzed in differentiating the A-EGJOO and F-EGJOO subgroups. RESULTS: The most common symptom of EGJOO was chest pain or chest discomfort (30.63%), then the dysphagia (29.73%), and acid regurgitation/heartburn (27.03%). Non-erosive reflux disease (36.04%) was the most popular cause for EGJOO, then the reflux esophagitis (17.12%). Besides the intra-EGJOO and extra-EGJOO lesions, the connective tissue disease (6.31%) and central nervous diseases (2.70%) were found to be the etiology of EGJOO. The causes of the rest 19 EGJOO were unknown. A-EGJOO patients presented significantly higher intra bolus pressure (IBP) than that of F-EGJOO [6.80 (5.20, 9.20) mmHg vs. 5.10 (3.10, 7.60) mmHg, P=0.016]. The area under curve of IBP was 0.637. When IBP≥5.15 mmHg, the sensitivity was 78.60% and specificity 50.70% to differentiate A- or F-EGJOO. CONCLUSION Chest pain or chest discomfort was the most common symptom in EGJOO patients. Besides the intraluminal structural disorders, the extra-luminal causes were found in EGJOO patients. A-EGJOO presented higher IBP than that of F-EGJOO patients. The cutoff value of IBP to differentiate A-EGJOO from EGJOO was 5.15 mmHg with sensitivity 78.06% and specificity 50.70%. However for the low area under curve, the diagnostic value of IBP was limited.
Deglutition Disorders ; Esophageal Motility Disorders/diagnosis* ; Esophagogastric Junction ; Humans ; Manometry ; Retrospective Studies

A new benzaldehyde, 3-hydroxy-4-(4-(2-hydroxyethyl) phenoxy) henzaldehyde(1), together with six known compounds, including isovanillic acid(2), pyrocatechol(3), glutinosalactone A(4), chrysoeriol(5), apigenin(6) and luteolin(7) were isolated from aerial part of Rehmannia glutinosa. The compounds were isolated by macroporous resin, silica gel, Sephadex LH-20 and HPLC chromatographies. The chemical structures of 1-7 were elucidated on the basis of spectral analysis (MS, 1D NMR and 2D NMR).
Benzaldehydes ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Rehmannia ; chemistry ; Spectrometry, Mass, Electrospray Ionization

In this paper, three kinds of chemotactic parameters (concentration, temperature and pH) were determined by plate assay and spore germination method to research the chemotactic response of Botrytis cinerea and Alternaria panax, and their spores on total ginsenosides. The results showed that Botrytis cinerea had strong chemotactic response at the mid-concentration of total ginsenosides (cultivation temperature was 20 degrees C and pH value was 6), and the data of chemotactic migration index (CMI) was 1.293 0, chemotactic growth rate (CGR) was 0.476 0, spore germination rate (SGR) was 53%, and dry weight of mycelial (DWM) was 0.452 6 g x L(-1); however, Alternaria panax had strong chemotactic response at the low-concentration of total ginsenosides (cultivation temperature was 25 degrees C and pH value was 6), and the data of chemotactic migration index (CMI) was 1.235 4, chemotactic growth rate (CGR) was 0.537 0, spore germination rate (SGR) was 67%, and dry weight of mycelial (DWM) was 0.494 8 g x L(-1). The results indicated that the low and middle concentration (2, 20 mg x L(-1)) of total ginsenosides had significant promoting effect on chemotactic response of these two pathogens, and the spore germination, mycelial growth rate, dry weight of mycelial of them were also significantly improved by this chemotactic response, whereas it decreased as the increase of total ginsenosides concentration.
Alternaria ; drug effects ; growth & development ; physiology ; Botrytis ; drug effects ; growth & development ; physiology ; Chemotaxis ; drug effects ; Drugs, Chinese Herbal ; metabolism ; pharmacology ; Ginsenosides ; metabolism ; pharmacology ; Panax ; metabolism ; microbiology ; Plant Diseases ; microbiology ; Spores, Fungal ; drug effects ; growth & development ; physiology

Objective To analyze the value of plane QRS-T angle on prediction of malignant ventricular arrhythmia (MVA) occurred after emergency percutaneous coronary intervention (PCI) in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods The clinical data of 418 patients with STEMI who underwent PCI within 12 h of symptom onset were retrospectively analyzed, and the patients were divided into plane QRS-T angle ≤ 90° group (324 cases) and plane QRS-T angle>90° group (94 cases) according to the plane QRS-T angle after PCI. The clinical data were compared between 2 groups. Results Compared with patients in plane QRS-T angle ≤ 90° group, patients in plane QRS-T angle > 90° group was older: (67.4 ± 11.8) years vs. (63.6 ± 12.0) years, QTc interval was longer: (438.60 ± 34.97) ms vs. (425.24 ± 25.49) ms, rate of left ventricular ejection fraction (LVEF) <45% was higher: 57.4% (54/94) vs. 35.8% (116/324), rate of using of beta-blockers was less: 74.5% (70/94) vs. 84.9% (275/324), but the incidences of hypertension and MVA were higher:79.8%(75/94) vs. 64.5%(209/324) and 10.6%(10/94) vs. 1.2%(4/324), and there were statistical differences (P<0.01 or<0.05). Logistic regression analysis showed that plane QRS-T angle >90° was an independent risk factor of MVA after PCI in STEMI patients (OR = 9.640, P =0.001), and using of beta-blockers was a protective factor (OR = 0.266, P = 0.028). Conclusions Plane QRS-T angle>90° is an independent risk factor of MVA after PCI in STEMI patients, while the use of beta-blockers is a protective factor. Paients with STEMI after PCI should be alert to the occurrence of MVA in the condition of plane QRS-T angle>90° and not taking beta-blockers.

Objective To evaluate the efficiency of rotational atherectomy (RA) with drug-eluting stents (DES) by intravascular ultrasound (IVUS) for elderly patients with heavily calcified lesions. Methods Thirteen patients aged above 70 years with heavily calcified coronary artery having underwent the treatment of RA and DES by IVUS were enrolled. The levels of troponin T, creatine kinase isozyme-MB and creatinine were detected before and after operation.The levels of minimus lumen diameter, diameter stenosis rate, effective area of the lumen before and after operation were compared. The immediate success rate was evaluated and the follow-up results were recorded. Results Thirteen patients in accordance with angiography and IVUS examination were confirmed as severe calcification. The levels of troponin T, creatine kinase isozyme-MB and creatinine detected before and after operation had no significantly differences (P>0.05). The levels of minimus lumen diameter, diameter stenosis rate and effective area of the lumen before and after operation had significant differences:(2.06 ± 0.38) mm vs. (3.98 ± 0.76) mm, (73.26 ± 7.02)%vs. (17.00 ± 3.34)%, (4.53 ± 1.50) mm2 vs. (12.54 ± 6.19) mm2, P<0.01. The rate of left anterior descending artery, circumflex artery and right coronary artery calcification was 10/13, 0 and 1/13. All patients′operation was successful. Followed up for (14.6 ± 3.4) months, no patients had angina and myocardial infarction, or required target vessel revascularization. Conclusions RA with DES by IVUS can be safely used in elderly patients with severely calcified coronary artery disease, and can improve the success rate of intervention operation.