We developed the monoclonal antibodies against nucleoprotein (NP) of Newcastle disease virus (NDV),and established a double antibody sandwich ELISA method for quantitative determination of NP antigen of NDV (NDV NP ELISA).The recombination NP protein derived from strain F48E9 of NDV were prepared and used to immunize BLAB/c mice.The mouse splenic cells from immunized mice were fused with SP2/0 cells to generate monoclonal antibodies (mAb).The NDV NP specific mAbs were paired to establish a double antibody sandwich ELISA method.The performance of the NDV NP ELISA was evaluated,including specificity,sensitivity,precision,accuracy and linearity.The correlation between the ELISA and PFU virus titer was analyzed by regression analysis method.Two monoclonal antibodies 3C10 and 4E7 were selected to establish double antibody sandwich ELISA for NP antigen of NDV.The linearity and performance of the NDV NP ELISA was characterized.The detection linearity fell in the range of 0.015-0.250 μg/mL (R2 =0.997 4).The detection limit of the assay was 0.015 μg/mL.The recovery was between 88.4％ and 106.01％;the variation coefficient was below 3.4％.In testing of 50 NDV virus samples,this assay performed well and correlated comparably with PFU virus titer (R2 =0.920 9).The NDV NP ELISA for quantitative detection of NDV is a reliable quantifiable assay for detection of NDV NP protein;it provides a new approach for rapid and quantitative detection of Newcastle disease virus.

Objective To evaluate the effect of injection of ozone (O3) through lateral recess for the treatment of radiculitis caused by lumbar disc herniation on the blood glucose in patients with diabetes mellitus.Methods Ninety-six patients with radiculitis caused by lumbar disc herniation involving 48 patients with diabetes mellitus and 48 patients without diabetes mellitus were enrolled in the study.The diabetic or non-diabetic patients were randomly divided into 2 groups ( n =24 each):non-diabetic patients-O3 group (group N-O3 ),non-diabetic patients-ghcocorticoid group (group N-GC),diabetic patients-O3 group (group D-O3 ),and diabetic patients-glucocorticoid group (group D-GC).In N-O3 and D-O3 groups,30 mg/L O3 10 ml was injected via the lateral recess.In N-GC and D-GC groups,compound betamethasone injection 3.5 mg was injected via the lateral recess.The blood glucose level was measured before treatment (T1),1 and 4 h after treatment (T3,4),and 1,3 and 7 days after treatment (T5-7).VAS score was recorded at T1,immediately after treatment (T2),and at T5-7.The patients' quality of life was measured using the Medical Outcomes Study 36-Item Short-Form Health Survey (MOS SF-36 Health Survey) questionnaire at T1,7.The therapeutic effect was evaluated at T7.The side effects were recorded.Results Compared with the baseline value at T1,VAS scores were significantly decreased after treatment,while the MOS SF-36 Health Survey questionnaire scores for physical functioning,bodily pain,role emotional,and mental health in all groups,for general health in group N-GC,and for vitality in groups D-O3 and N-O3 were significantly increased at T7,and the blood glucose level was significantly increased at T4 in groups N-GC and D-O3,and at T3-7 in group D-GC ( P ＜ 0.05).There was no significant change in the blood glucose level before and after treatment in group N-O3.The blood glucose level was significantly higher in group N-GC than in group N-O3,and in group D-GC than in group D-O3 ( P ＜ 0.05).There were no significant differences in the excellent and good rates and all the MOS SF-36 Health Survey questionnaire scores among all groups.No side effects were found in the study.Conclusion Injection of ozone through lateral recess for treatment of radiculitis caused by lumbar disc herniation exerts no influence on the blood glucose level in patients with diabetes mellitus.

Objective: The aim was to explore the association between the HLA-DRB1 allele polymorphism and susceptibility to chronic myelogenous leukemia(CML)in Chinese Han population.Methods: The polymorphism of HLA-DRB1 alleles in 762 CML patients and 2 264 normal umbihcal cord blood samples were tested by polymerase chain reaction with sequence specific primer(PCR-SSP).Results: The most common allele in DRB1 locus in normal population was DRB1 * 15(17.25%), whereas DRB1 * 10 was the least frequent allele(1.39%).Compared with normal controls, the frequency of HLA-DRB1 * 08 was significantly increased in all CML groups(7.48%vs 5.39%, X~2 =8.963,OR= 1.023, P =0.004)and CML male groups(7.72%vs 5.39%,X~2 = 8.059,OR= 1.025,P = 0.007).But there was no significant difference between the normal controls and CML female patients.Conclusion: It should be concluded that DRB1 * 08 could be considered a susceptible allele for CML male patients.

Objective To evaluate the utility of short tau inversion recovery (STIR) sequence in the diagnosis of hippocampal sclerosis ( HS).Methods Twenty-one patients with medial temporal lobe epilepsy without neoplasm lesions or injuries by conventional MRI sequence including T1WI,T2WI and FLAIR were included in this study.STIR imaging in axial,coronal and sagittal sequences was performed on these patients. Diagnosis of HS was based on the findings of hippocampal atrophy,alteration signal,disturbed internal structure and enlargement of the inferior horn. The findings shown on conventional MRI were compared with those on STIR sequence. Furthermore,the correlation of radiologic and histological findings was investigated in 6 patients operated for refractory seizures. Results On conventional MRI sequence,14 patients (66.7％) were confirmed with unilateral HS and 4 patients were suspected with unilateral HS. In contrast,all these 18 patients (85.7％) were confirmed with unilateral HS by STIR.Particularly,STIR sequence delineated the internal structure of hippocampus more clearly than conventional MRI sequences did. C shaped contour in subiculum-CA1-CA2 was revealed in normal hippocampus on STIR sequence and disappeared in HS,correlated to the pathology finding of loss of neuron in CA1 in resected tissues in 6 operated patients.The patients with HS also showed areas of hypodensity in CA4 on STIR,in accordance with pathologic findings of gliosis in this area in the 6 operated patients.Conclusion STIR sequence could depict the internal anatomical structure of hippocampus with high resolution superior to conventional MRI sequences,and can be of great value in the diagnosis of HS.

Objective To investigate the protective effects of isopimaric acid ( ISO), the BKCa channel activator, on cognitive function and synaptic plasticity in APP/PS1 mice. Methods Alzet osmotic pump was loaded with ISO or DMSO only and assembled with ALZET Brain Infusion Kit III. The cannula was implanted into the lateral ventricle of 4-month-old male APP/PS1 mice or matched wild type ( WT) mice. Two weeks later, open field test and Morris water maze were conducted. Paired-pulse facilitation ( PPF) and TBS-induced long-term potentiation ( LTP ) were recorded in CA1 region of hippocampus. Results The open field test showed that there was no significant difference among the four groups in spontaneous activities and vertical plane movement distance within 30 minutes. Floor plane movement distance was significantly greater in APP/PS1+DMSO group than that in WT+DMSO group(P<0.05) . Compared with the WT+DMSO group, APP/PS1+DMSO group had significantly longer escape latency from the third to fifth day and lower percentage of time spent in the target quadrant ((43.27±3.24)% vs (34.19±2.56)%) and the number of crossing through the platform ((4.25±0.66)times vs (1.93±0.33)times)(P<0.05). Compared with the APP/PS1+DMSO group, the APP/PS1+ISO group had significantly shorter escape latency from the fourth to fifth day and higher percentage of time spent in the target quadrant ((46.16±3.51)%) and the number of crossing through the platform ((3.41±0.34) times) (P<0.05). PPF in APP/PS1+DMSO group significantly reduced compared with that in WT+DMSO group at 30-50ms interstimulus interval(P<0.05). PPF in APP/PS1+ISO group((224.50±13.79)%) was significantly augment compared with APP/PS1+DMSO ((174.99 ±6.68)%) group at 40 ms interstimulus interval (P<0.05). The LTP at 60 min post-TBS was significantly smaller in the APP/PS1+DMSO group ((135.19±1.32)%) than that in the WT+DMSO group ((172.17± 4.15)%)(P<0.001). The LTP of the APP/PS1+ISO group((160.48±1.19)%) became significantly in-creased compared with that in the APP/PS1+DMSO group(P<0.001).Conclusion BKCa channel activator ISO improve the learning and memory function of APP/PS1 mice by promoting PPF and increasing LTP to recover synaptic plasticity in the hippocampus.

Objective To investigate the change in NF-κB activity in astrocytes in spinal dorsal horn in a rat model of neuropathic pain and the underlying mechanism. Methods Sixteen male SD rats aged 2-3 months weighing 220-280 g were randomly divided into 2 groups ( n = 8 each) : sham operation group (group S) and CCI group. Neuropathic pain was induced by chronic constrictive injury (CCI) . Right sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4-0 chromic catgut. In group S the right sciatic nerve was exposed but not ligated. The paw withdrawal threshold (PWT) to von Frey filament stimulation and paw withdrawal latency (PWL) to radiant heat stimulation were measured at 1 d before (baseline) and 7 d after operation. The animals were then killed and the lumbar segment of the spinal cord (L_(4-6)) was removed. The expression of NF-κB in the astrocytes in spinal dorsal horn was determined by immuno-histochemistry. Results PWT and PWL to mechanical and thermal stimuli were significantly decreased after operation as compared with the baseline before operation in group CCI. The number of NF-κBp65 immunoreaction positive cells in the spinal dorsal horn on the operated side was significantly larger in group CCI than in group S. Conclusion NF-κB signal transduction pathway in the astrocytes in the spinal dorsal hom may be involved in neuropathic pain.

Objective To explore a new index for reflecting the topological information of brain functional networks in patients at high risk of Alzheimer disease using characteristics of resting-state functional connectivity strengths(FCS) in patients with amnestic mild cognitive impairment(aMCI). Methods Thirty-one aMCI patients and 42 age, gender and years of education matched normal controls were enrolled between September 2009 and April 2011 in this study. The resting-state functional MRI (rs-fMRI) data of all participants were acquired and preprocessed. Then the whole-brain functional connectivities were constructed for exploring the distribution characteristics of hub regions which had higher FCS values. Using two-sample t test to compare group differences in age, years of education and each neuropsychological assessment. In addition, using Chi-squared test to compare group differences in gender. Group differences in FCS values were analyzed by general linear model. Finally, correlation analyses were used to evaluate the relationships between the FCS values of the brain regions with group differences and behavioral scores in aMCI patients. Results The hub regions of the functional networks in the aMCI patients were mainly located in the association cortices such as the precuneuses, posterior cingulate cortices, medial prefrontal cortices, angular gyri, superior occipital gyri, fusiform gyri and lingual gyri. The distribution models in the aMCI patients were consistent with those in the normal controls. However, the FCS values of these brain regions were significantly lower in the aMCI patients than those in the normal controls. In comparison to the normal controls, the aMCI patients had significantly decreased FCS values in the bilateral fusiform gyri, lingual gyri, superior occipital gyri, left middle occipital gyrus and postcentral gyrus (the cluster was 389, 230, 187 and 107 voxels, respectively;P<0.05, respectively), and they had decreased trends of FCS values in the bilateral posterior cingulate cortices and right insulas. The correlation analysis with uncorrected conditions showed that the FCS values of the left postcentral gyri were correlatid with the clock drawing test (CDT) scores (r=0.436, P=0.026). Conclusions aMCI mainly attacks the hub regions of brain functional networks. The changes of functional connectivities in aMCI may reflect the early pathophysiologic alterations of AD.

Objective:To evaluate the value of 99Tc m-methoxyisobutylisonitrile(MIBI) SPECT/CT imaging for the identification of dystonic muscles in patients with primary cervical dystonia (PCD). Methods:A total of 10 patients with PCD (3 males, 7 females, age (47.3±9.9) years) and 10 healthy subjects (4 males, 6 females, age (43.5±9.4) years; control group) between August 2019 and October 2021 in China-Japan Friendship Hospital were enrolled prospectively. All subjects underwent 99Tc m-MIBI SPECT/CT scan. The SUV max of 8 bilateral representative muscles, including rectus capitis posterior major, obliquus capitis inferior, splenius capitis, semispinalis, sternocleidomastoid, trapezius, musculus scalenus muscle and levator scapulae were evaluated in control group. In PCD group, muscles with abnormal uptake were determined. ROI was drawn and SUV max was measured. Independent-sample t test was used to analyze the differences of SUV max between normal and abnormal muscles. The detecting rates of neck MRI and SPECT/CT for abnormal muscles were analyzed by χ2 test. Results:Normal muscles of healthy subjects showed mild symmetrical radioactivity distribution, with the SUV max of 1.10±0.19. A total of 60 muscles with abnormal uptake in 10 patients were found, including 7 rectus capitis posterior major, 10 obliquus capitis inferior, 8 splenius capitis, 8 semispinalis, 10 sternocleidomastoid, 5 trapezius, 3 musculus scalenus muscle and 9 levator scapulae. The SUV max of muscles with abnormal uptake was 1.81±0.43, which was higher than that of normal muscles ( t=17.05, P<0.001). Only 30 pieces abnormal hypertrophy muscle were found by neck MRI, and the detecting rate was much lower than that of SPECT/CT (18.75%(30/160) vs 37.50%(60/160); χ2=28.03, P<0.001). Conclusion:99Tc m-MIBI SPECT/CT may be a useful method for identifying dystonic muscles and a guide to precision therapy in patients with PCD.

OBJECTIVETo explore the promoting effects of cograft of auto-microskin and allo-dermal matrix on burn wound healing.

METHODSForty-six patches of full skin loss wounds were made in the backs of 6 small white swines. Auto-microskin and meshed acellular allo-dermal matrix were simultaneously prepared. The 46 wounds were averagely divided into test (T) and control (C) groups. The wounds in T group were covered with auto-microskin and allo-dermal matrix (1:4) and split-thick allo-porcine skin. While the wounds in C group were grafted with autoskin and allo-dermal matrix.

RESULTSThe skin survival rate exhibited no difference between C and T groups (P > 0.05). There was similar histological exhibition in the two groups after skin grafting. The structure of collagen fibres appeared integrated, clear with regular arrangement and steady diameter at 8 - 20 post-operative weeks. Simultaneously, it was revealed by histological examination that normal vascular structure could be identified in the grafted skin and that inflammatory reaction ameliorated gradually and that epithelium combined well with dermis. It was also found that the epidermal papillar across the basement membrane fixed well to allo-dermis. The skin appeared smooth, elastic and functioned well at 5 months after skin grafting.

CONCLUSIONThe grafted skin survived well after the cograft of auto-microskin and allo-dermal matrix, which might be ideal covering material for the major deep burn wound healing.

Animals ; Burns ; therapy ; Disease Models, Animal ; Female ; Graft Rejection ; Male ; Skin Transplantation ; Skin, Artificial ; Swine

Sporotrichosis has been classified as "neglected tropical diseases" by the World Health Organization. In recent years, researches on sporotrichosis have gradually increased. This review summarizes recent progress in sporotrichosis in terms of pathogens, epidemiology, diagnosis, treatment and vaccines.