To study the B cell linear epitopes of rabies virus CVS-11 nucleoprotein, peptides were synthesized according to the amino acid sequences of B cell linear epitopes. Linear epitopes predicted by bioinformatics analysis were evaluated with immunological techniques. Indirect enzyme-linked immunosorbent assay showed that titers of antibodies to peptides (355-369 and 385-400 residues of rabies virus CVS-11 nucleoprotein) were above 1:12 800 in mouse sera. The antibodies recognized denatured rabies virus CVS-11 nucleoprotein in Western blot analysis. Purified anti-peptide antibodies recognized natural rabies virus CVS-11 nucleoprotein in BHK-21 cells in indirect fluorescent antibody test. The 355-369 and 385-400 residues of rabies virus CVS-11 nucleoprotein were validated as B cell linear epitopes.
Amino Acid Sequence ; Animals ; Antibodies, Viral ; immunology ; Epitope Mapping ; Epitopes, B-Lymphocyte ; chemistry ; genetics ; immunology ; Female ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Nucleoproteins ; chemistry ; genetics ; immunology ; Rabies ; immunology ; virology ; Rabies virus ; chemistry ; genetics ; immunology

Objective To evaluate the effectiveness and safety of mouse nerve growth factor in treating dysphagia in patients with nasopharyngeal carcinoma after radiotherapy.Methods Fifty-eight post-radiotherapy nasopharyngeal carcinoma patients with dysphagia were randomly divided into an observation group and a control group.Both groups received routine treatment,but the observation group was additionally injected with mouse nerve growth factor intramuscularly every day for four weeks.Before and after the 4 weeks of treatment,both groups were evaluated using Kubota's water drinking test,videofluoroscopy and the brief version of the WHO's Quality of Life scale.Results After 4 weeks,the patients in the observation group displayed significantly greater improvement in swallowing compared with the control group.There was a significant difference in the groups' average scores on the drinking water test and in the videofluoroscopy results.Moreover,the patients in the observation group had significantly higher quality of life scores than those in the control group,on average.Conclusions Mouse nerve growth factor may have a rapid and safe therapeutic effect on dysphagia induced by radiation.No obvious adverse reactions were observed.

Objective To investigate the effect and mechanism of cell differentiation agent Ⅱ (CDA-Ⅱ) on the differentiation of human acute mycloid leukemia (APL) HL-60 cells.Methods The cell morphology and differentiation was detected by Wright-Giemsa staining,the expression of cell surface differentiation antigen CD11b of HL-60 was detected by flow cytometry,the differentially expressed genes were screened by gene expression profile chip (NimbleGen).Results The result of Wright-Giemsa staining showed that CDA-Ⅱ induced HL-60 differentiation towards mature stages in a time-dependent manner.After treated with CDA-Ⅱ,the percentage of CD11b-positive HL-60 cells significantly increased in a time-and dose-dependent manner.The result of gene expression profiling indicated differentially expressed genes including 113 up-regulation genes and 140 down-regulation genes.The up-regulation expression genes involved in six pathways including mineral absorption,complement and coagulation cascades,down-regulation expression genes involved in nine pathways including pyrimidine metabolism,RNA polymerase,purine metabolism and so on.Conclusion CDA-Ⅱ can induce HL-60 differentiation and make gene differentially expressed.The data provide the feasibility of CDA-Ⅱ in differentiation induction therapy for APL.

Objective To investigate the safety of cleaning and disinfection of duodenoscope used in endoscopic retrograde cholangiopancreatography (ERCP).Methods The duodenoscopes,which were used in Endoscopy Center of Digestive Disease Hospital,Heilongjiang Provincial Hospital from May 4 to May 31 2016,were cleaned manually and disinfected with sterilization powder.The samples from elevator and elevator channel were collected for bacterial culture to evaluate the safety of clinical application of duodenoscope.Results Among 60 high level disinfections,one was excluded because disinfectant concentration wasn't effective.Among 59 elevator samples no positive cultures were detected,while among 59 channel samples,4(6.8％) cultures were positive and all from the same duodenoscope,including 3 cases of Klebsiella pneumonia and 1 case of Pseudomonas aeruginosa.Conclusion Sterilization powder is effective for disinfection of duodenoscope used for ERCP.Residual organic material can't be excluded from elevator sample even if it is negative for culture.Duodenoscope with high level disinfection still has the risk of transmitted infection,and should be monitored regularly by culture and reprocessed in time.

Objective:To investigate the expression of serum human phosphatidylethanolamine-binding protein 4 (hPEBP4) in patients with multiple myeloma (MM) and its clinical significance.Methods:A total of 59 symptomatic MM patients admitted to West Branch of Beijing Chaoyang Hospital from September 2016 to September 2018 were selected as the research objects. According to the CRAB symptoms [elevated serum calcium (C), kidney injury (R), anemia (A), bone lesions (B)], all patients were divided into 2 groups, including the active group of 44 patients with CRAB symptoms, and the response group of 15 patients who achieved at least partial remission after chemotherapy and symptom relief of CRAB. According to the degree of bone lesions (BL), 30 patients with severe bone-related events were grouped as the severe bone lesions (SBL) group, and 14 patients were grouped as the non-severe bone lesions (NSBL) group. According to the revised international prognostic staging system (R-ISS), patients in the active group were divided into three subgroups: stage Ⅰ, stage Ⅱ, and stage Ⅲ, including 26, 11 and 7 patients, respectively. A total of 15 healthy examination people whose gender and age matched those of the patients were treated as the healthy control group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of hPEBP4, tumor necrosis factor ligand superfamily member 14 (LIGHT/TNFSF14) and activin A of patients in different groups. Pearson was used to analyze the relationship of the expressions of multiple factors in the active group. The optimal cut-off value of multiple factors diagnosing MM was determined by using receiver operating characteristic (ROC) curve, and according to the cut-off value, the differences in overall survival (OS) of patients with different stratification were compared.Results:In the active group, the respond group, the healthy control group, the level of hPEBP4 was (1.48±0.64) μg/L, (1.49±0.75) μg/L, (0.31±0.10) μg/L, respectively; the level of LIGHT/TNFSF14 was (169±112) ng/L, (256±132) ng/L, (44±27) ng/L,respectively; the level of activin A was (383±266) ng/L, (223±79) ng/L, (234±85) ng/L, respectively; and the differences were statistically significant (all P<0.05). In the active group, the level of hPEBP4 was (1.06±0.60) μg/L, (1.15±0.50) μg/L, (1.73±0.68) μg/L, respectively in patients with stage R-ISSⅠ, R-ISSⅡ and R-ISS Ⅲ, and the difference was statistically significant ( F=3.287, P=0.032). The level of activin A was (219±55) ng/L, (247±117) ng/L, (450±215) ng/L, respectively among patients in stage R-ISSⅠ, R-ISSⅡ, R-ISS Ⅲ, and the level of activin A in stage R-ISS Ⅲ was higher than that in stage R-ISSⅠand R-ISSⅡ (all P < 0.05). The levels of LIGHT/TNFSF14 and activin A of SBL patients were higher than those of NSBL patients [(174±101) ng/L vs. (98±53) ng/L; (467±238) ng/L vs. (189±71) ng/L, all P < 0.05]. The level of hPEBP4 was positively correlated with the levels of M protein ( r=0.694, P < 0.01) and activin A ( r=0.252, P < 0.01) of IgG patients in the active group. ROC curve analysis showed that the optimal cut-off value of hPEBP4, LIGHT/TNFSF14, activin A diagnosing MM was 1.04 μg/L, 97.0 μg/L, 156.2 ng/L. The median overall survival (OS) time of patients with hPEBP4 >1.04 μg/L and hPEBP4 ≤ 1.04 μg/L was 57 months (95% CI 22-92 months) and not reached, respectively, and the difference was statistically significant ( P < 0.05); while the median OS time of patients with activin A ≥ 156.2 ng/L and activin A < 156.2 ng/L was 61 months (95% CI 24-98 months) and not reached, respectively, and the difference was statistically significant ( P < 0.05). Conclusions:High expression level of hPEBP4 is related with the progression of MM. It is positively related with the level of M protein and negatively with the OS of MM patients. It is suggested that hPEBP4 may be used as an important marker to judge disease progression and tumor burden in MM. LIGHT/TNFSF14 and activin A cooperate with hPEBP4 to participate in the pathological processes of tumor microenvironment of MM.

Objective To establish a rapid fluorescent inhibition test (RFFIT) for testing rabies virus neutralizing antibody and the titer of rabies virus neutralizing antibody.CVS-11 was used as the standard challenge virus,and three generations prepared for the establishment of the virus library.Methods International standard for rabies immunoglobulin was used as the reference serum.RFFIT test was established under consulting the protocol of Institute of Pasteur,and its specificity,stability and reproducibility were validated.Results We established the RFFIT which showed both good specificity ( 100% ) and reproducibility (P＞0.5).Conclusion The establishment of RFFIT test perfected the rabies laboratory techniques and would enhance the overall ability in detecting rabies in China.

OBJECTIVECompare the difference of the results referred to the WHO standard rabies immunoglobulin and the national standard human rabies immunoglobulin used in the rapid fluorescent focus inhibition test (RFFIT).

METHODSSetting the WHO standard immunoglobulin and the national standard immunoglobulin in the same system and testing 12 human serum at the same time. Compare the fluorescence percentage of the two different standard immunoglobulin; compare the 12 serum results calculated from the two different standard immunoglobulin used the calculation formula of neutralization antibody titer.

RESULTSThe Results display that the 50% percent of the two standard immunoglobulin are all between the fifth and the sixth well, but the percentage of the national standard immunoglobulin is lower than the WHO one. The same testserum result calculated from the WHO standard immunoglobulin is little higher than the national one.

CONCLUSIONThere is difference in the WHO standard immunoglobulin and the national one, but there is no influence in the results.

Fluorescent Antibody Technique ; methods ; standards ; Humans ; Immunoglobulins ; immunology ; Neutralization Tests ; methods ; standards ; Rabies ; diagnosis ; immunology ; Rabies virus ; immunology ; Reference Standards ; World Health Organization

BACKGROUND:Pain is the main clinical manifestation for ankylosing spondylitis. At present, nonsteroid anti-inflammatory drugs are oral y taken, but the effects are limited and toxic and side effects are more. Thus, there is no effective scheme for treatment of pain induced by ankylosing spondylitis.
OBJECTIVE:To investigate the correlation between postoperative joint pain al eviation and al ogeneic blood transfusion, and the mechanisms.
METHODS:We retrospectively analyzed clinical data of 88 ankylosing spondylitis patients combined with kyphosis who received only one section of osteotomy surgery merging hip joint pain. We compared the visual analog scale score of hip joint and detected the variation of leucocytes, lymphocytes and immunoglobulin concentrations before and after the operation in the groups of fresh al ogeneic whole blood transfusion, autologous whole blood transfusion, and mixed transfusion of al ogeneic and autologous whole blood. Flow cytometry was used to analyze the number and ratio of peripheral blood Th17 cells and Treg cells which were both highly associated with autoimmune diseases.
RESULTS AND CONCLUSION:The symptom of hip arthralgia obviously improved in both groups transfused by fresh al ogeneic whole blood or al ogeneic-autologous mixed whole blood. However, no obvious variation was detected in leucocytes, lymphocytes and immunoglobin concentration. However, flow cytometry results indicated that Th17/Treg proportion associated with autoimmune diseases was increased remarkably in peripheral blood of ankylosing spondylitis patients. Results suggested that al ogeneic whole blood transfusion can al eviate patients’ joint pain by correcting the imbalance of Th17/Treg which may improve their immune state.

The combination of Danggui and Honghua (GH) is a popular herb pair commonly used in clinic for the treatment of blood stasis syndrome in China. To evaluate the activating blood circulation and dissipating blood stasis effects of the combination of different proportions of Danggui and Honghua on acute blood stasis rats, and optimize the proportion of GH to have the best activating blood circulation and dissipating blood stasis effect. Acute blood stasis rat model was induced by subcutaneous injection of adrenaline and ice water bath. The blood stasis rats were administrated intragastrically with GH (1 : 0, 4 : 1, 2 : 1, 3 : 2, 1 : 1, 2 : 3, 1: 2, 1 : 4 and 0 : 1) extracts. The whole blood viscosity (WBV), plasma viscosity (PV), and high shear whole blood relative index (HSWBRI), low shear whole blood relative index (LSWBRI), and erythrocyte aggregation index (EAI) were tested to observe the effects of GH on hemorheology of blood stasis rats. And the maximum aggregation induced by adenosine diphosphate (ADP) was tested to observe the effect of GH on platelet aggregation index of blood stasis rats. In addition, the prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT) and plasma fibrinogen (FIB) were tested to observe the effects of GH on blood coagulation function of blood stasis rats. Then principal component analysis and multi-attribute comprehensive index methods were both used to comprehensively evaluate the total activating blood circulation and dissipating blood stasis effects of GH. The results showed that the hemorheological indexes and coagulation parameters of model group both had significant differences with normal group. Compared with model group, GH (1 : 0, 4 : 1, 2: 1, 3 : 2, 1 : 1, 2 : 3, 1 : 2, 1 : 4 and 0 : 1) could improve all the blood hemorheology indexes and regulate part indexes of blood coagulation function and platelet aggregation in acute blood stasis rats. Based on principal component analysis and multi-attribute comprehensive index methods, GH 1 : 1 and GH 3 : 2 both had the best effect of blood circulation and dissipating blood stasis, and the effect of GH 1 : 1 was slightly better than GH 3 : 2. These results suggest that GH could obviously ameliorate the abnormality of hemorheology and blood coagulation function in acute blood stasis rats. The optimized proportion of GH was consistent with regulations of medicine usage that GH 1 : 1 had the highest frequency used in traditional Chinese formulae. It could provide scientific basis for more effective application of the compatibility between Danggui and Honghua in modern clinic medicine.
Animals ; Blood Coagulation ; Blood Viscosity ; Carthamus tinctorius ; China ; Drugs, Chinese Herbal ; pharmacology ; Erythrocyte Aggregation ; Hemorheology ; Partial Thromboplastin Time ; Platelet Aggregation ; Principal Component Analysis ; Prothrombin Time ; Rats ; Rats, Sprague-Dawley ; Thrombin Time

Objective To report a new self-designed locking plate assembly suitable for treatment of displaced calcaneal fractures of Sanders types Ⅱ &Ⅲ via sinus tarsi approach.Methods We designed a new locking plate assembly suitable for treatment of displaced calcaneal fractures of Sanders types Ⅱ & Ⅲ via sinus tarsi approach.Its biomechanical performance was tested by standard electronic and mechanical devices.From March 2014 to October 2015,18 patients with displaced calcaneal fracture were treated with our self-designed new locking plate assembly.They were 14 men and 4 women,aged from 32 to 66 years (average,50.4 years).All the fractures were unilateral closed ones,with 7 cases of the left side and 11 ones of the right side.By the Sanders classification,14 cases belonged to type Ⅱ (8 to type Ⅱa,4 to type Ⅱb and 2 to type Ⅱc) and 4 to type Ⅲ (2 to type Ⅲab,one to type Ⅲac and one to type Ⅲbc).The B(o)hler angle,Gissane angle,and height,width and length of the calcaneus were measured and compared at preoperation,postoperative 3 months and final follow-ups.The clinical outcomes were evaluated using Maryland foot score system atfinal follow-ups.Results This new locking plate assembly could tolerate a maximum vertical load of 1,396.03 N and a maximum horizontal anti-bending load of 427.15 N.It broke after it had been subjected to 93,003 loadings in a repeated 500 N stress test.All the 18 patients were followed-up for an average of 9.3 months (from 6.0 to 13.5 months).Follow-ups at 3 months postoperation showed significant improvements in B(o)hler and Gissane angles and calcaneal height,width and length compared with pre-operative parameters (P ＜ 0.05),but no significant differences were observed in the above parameters between 3 and final follow-ups postoperation (P ＞ 0.05).Maryland foot scores demonstrated 12 excellent cases,3 good ones and 3 fair ones.No incision infection,implant failure,nonunion or malunion happened in this series.Conclusions This new locking plate assembly suitable for sinus tarsi approach can be used in clinic because it has advantages of easy placement,rigid fixation,satisfactory functional recovery of the foot and limited complications.