Objective We increase the soluble expression of artificial tandem hybrid ubiquitin binding domains ( ThUBD) in prokaryotic cytoplasm of Escherichia coli BL21 ( DE3 ) , which offer an effective and special profiling for ubiquitin conjugates( UbC) .Methods Codon optimization of the ThUBD was performed, followed by analysis of codon relative adaptiveness based on relative frequency of synonymous codon ( RFSC) of E.coli.Further induced expression and yeast ubiquitin conjugate enrichment quantified the soluble ThUBD-S and tested the ability to bind UbC.Results The statistical result showed that the percentage of codon of the highest usage frequency was increased from 48%to 75%, and codon adaptation index( CAI) was increased from 0.63 to 0.88 after codon optimization, which might suggest a higher expression of the ThUBD in E.coli BL21 (DE3).The subsequent SDS-PAGE indicated that the soluble target protein was increased four times, which accounted for 13.06%of total cell lysis.Further ubiquitinated proteome of yeast demonstrated that the ability to bind and enrich UbC of optimized ThUBD-S did not change compared with original ThUBD.Conclusion The expression of ThUBD-S can quadruple after codon optimization.At the same time, codon optimization does not impact its soluble expression and the ability to bind UbC.

Objective To analyse clinical application of hepatitis B virus core antibody(HBcAb)detected by using the chemilu-minescence microparticle immunoassay.Methods A total of 1 6 830 specimen with positive HBcAb detected by using the two pairs of semi-hepatitis test from January 2012 to November 2014 were collected,and divided into three groups according to the cut off in-dex(COI)of detection results of HBcAb,including group 1.0-<9.0,group 9.0-<1 1.0 and group COI≥1 1.0,and detection re-sults were statistically analysed.The hepatitis B virus(HBV)DNA test was carried out in specimen with negative hepatitis B surface antigen(HBsAg)and hepatitis B surface antibody (HBsAb)and COI≥1 1.0.Results The detection rate of HBsAg(+)HBsAb(-) (13.84%)was significantly higher than other expression patterns in group ≥1 1.0(P <0.05).There was no statistically significant differences in positive rate among all expression patterns of HBsAg and HBsAb in the group 9.0-<1 1.0(P >0.05).The detec-tion rate of HBsAg(+)HBsAb(-)of group 9.0-<1 1.0 was significantly lower than that of the other two groups(P <0.05).A total of 304 specimen were HBsAg(-)HBsAb(-)and COI≥1 1,among them 64 specimen were HBV DNA postive and the posi-tive rate was 21.0%.Conclusion In the detection of HBcAb,COI≥1 1 and 1.0-<9.0 could be reference indicators for diagnosiing current and past HBV infection respectively,which should be combined with other laboratory indicators of HBV clinical data for comprehensive analysis.

Objective: To investigate the screen exposure status and influencing factors among 6-12 year-old children in Hainan Province, so as to provide insights into screen exposure intervention for children. Methods: Children aged 6-12 years from 18 counties (cities) in Hainan Province were selected using multi-stage stratified cluster sampling method from December 2020 to July 2021. Demographic information, parents' educational level, family type and screen time was collected using questionnaire surveys. The screen exposure rate of children was analyzed, and factors affecting screen exposure were identified using a multivariable logistic regression model. Results: A total of 27 501 children were surveyed, including 13 901 boys (50.55%) and 13 600 girls (49.45%). The mean age was (9.22±1.86) years. Among them, 3 925 children had screen exposure, with a screen exposure rate of 14.27%. Multivariable logistic regression analysis showed that gender (female, OR=0.859, 95%CI: 0.796-0.926), age (OR=1.078, 95%CI: 1.049-1.108), ethnicity (ethnic minorities, OR=1.147, 95%CI: 1.041-1.254), place of residence (rural area, OR=0.869, 95%CI: 0.801-0.944), father's educational level (high school or technical secondary school, OR=0.879, 95%CI: 0.788-0.981; college degree or above, OR=0.686, 95%CI: 0.589-0.818), mother's educational level (college degree or above, OR=0.706, 95%CI: 0.588-0.846), family type (others, OR=1.250, 95%CI: 1.105-1.414), and annual family income (>100 000 Yuan, OR=0.741, 95%CI: 0.619-0.885) were the influencing factors for screen exposure among children aged 6-12 years. Conclusion The screen exposure among children aged 6-12 years in Hainan Province was affected by gender, age, ethnicity, place of residence, parental education level, family type and annual family income.

OBJECTIVE: To investigate the characteristics of T lymphocytic subsets in chronic tonsillitis patients for evaluation of their clinical implication. METHOD: Fresh peripheral blood samples were obtained from 54 chronic tonsillitis patients and 52 healthy counterparts. CD4+ and CD8+ T lymphocyte subsets including the naive (CD45RA+), memory (CD45RO+), functional (CD28+), activated (HLA-DR+, CD25+) and apoptosis (CD95+) T lymphocytes, were analyzed by flow cytometry, respectively. The clinical data such as serum Cystatin C con centration, ASO and ESR were simultaneously recorded from each chronic tonsillitis patient. RESULT: The CD4+ T cells, the rate of CD4+/CD8+ and CD4+ CD45RA+ /CD4+ CD45RO+, the CD4+ CD45RA+ T cells and the CD4+ CD25+ T cells in chronic tonsillitis were significantly lower than those of the control group (P < 0.05) while an obviously increasing percentage of memory (CD45RO+) and apoptosis (CD95+) T lymphocytes in chronic tonsillitis patients, and there were significant differences between the patients with chronic tonsillitis and the healthy volunteers (P < 0.05). Furthermore, in the chronic tonsillitis patients, Serum Cystatin C level was negatively correlated with CD4+ CD45RA+ T(P < 0.05), and not significantly correlated with other T lymphocyte subtypes (P > 0.05). CONCLUSION Immune disorder is present in the peripheral blood of chronic tonsillitis patients. Our data may provide valuable information for evaluation of disease progression of chronic tonsillitis patients.
Adolescent ; Adult ; CD4-Positive T-Lymphocytes ; Case-Control Studies ; Chronic Disease ; Female ; Flow Cytometry ; Humans ; Leukocyte Common Antigens ; Lymphocyte Count ; Male ; Middle Aged ; T-Lymphocyte Subsets ; Tonsillitis ; blood ; physiopathology ; Young Adult

Objective:To investigate the effect of bone mesenchymal stem cells derived exosomes (BMSC-Exo) on improving hippocampal microangiogenesis, energy metabolism, and behaviors in depression mouse models.Methods:(1) Mouse bone marrow mesenchymal stem cells were isolated and cultured to extract BMSC-Exo; BMSC-Exo morphology was observed by transmission electron microscopy, BMSC-Exo particle diameter ranges were determined by Zetaview analyzer, and expressions of CD9 and CD63 in BMSC-Exo were detected by Western blotting. (2) Depression models were established in 2 mice by chronic unforeseeable mild stress (CUMS); 24 h after stereotaxic injection of phosphate buffer solution (PBS) or DiR labeled BMSC-Exo, BMSC-Exo uptake was detected by in vivo imaging system. (3) Thirty-six mice were randomly divided into control group, model group and BMSC-Exo group ( n=12); CUMS was used to establish depression models in the latter 2 groups; brain stereotaxic injection of 1 μL BMSC-Exo was given to mice in the BMSC-Exo group after modeling, and same amount of PBS was given to the control group; behaviors were observed by forced swimming test (FST), tail suspension test (TST) and open field test (OFT); hippocampal microvascular length and number were detected by alkaline phosphatase staining; energy metabolism in the hippocampus was detected by micro positron emission tomography/computed tomography (mPET/CT); glucose transporter 1 (GLUT1) expression in the hippocampus was detected by Western blotting. Results:(1) BMSC-Exo had a typical disk-like vesicle-like structure with particle size of (100.5±1.4) nm; Western blotting confirmed that CD9 and CD63 expressed in BMSC-Exo. (2) In vivo imaging showed no fluorescence in the brain and liver after PBS injection, but obvious local fluorescence after BMSC-Exo injection. (3) Compared with the control group, the model group and BMSC-Exo group had significantly longer rest time in FST and TST and shorter movement distance and time in the central region of OFT ( P<0.05); compared with the model group, BMSC-Exo group had significantly shorter rest time in FST and TST and longer movement distance and time in the central region of OFT ( P<0.05). Compared with the control group, the model group and BMSC-Exo group had significantly decreased standard uptake value (SUV) of regions of interest, microvascular length and number, and GLUT1 expression in the hippocampus ( P<0.05); compared with the model group, the BMSC-Exo group had significantly higher SUV, microvascular length and number, and GLUT1 expression in the hippocampus ( P<0.05). Positive correlations were noted between hippocampal microvascular length and SUV and between microvascular number and SUV in the 3 groups ( r=0.540, P<0.001; r=0.600, P<0.001). Conclusion:BMSC-Exo could promote microangiogenesis energy metabolism in the hippocampus to improve depression-like behaviors in depression mouse models.

OBJECTIVE: To evaluate the treatment responses of persistent allergic rhinitis with and without eosinophils (EOS) in nasal discharge to inhaled glucocorticosteroid (CS), and therefore to verify whether low eosinophil level in nasal discharge can predict poor response to treatment with CS. METHOD: Forty-two symptomatic allergic rhinitis patients, who had not received CS therapy in three months preceding the study, were examined before and 2 month, 4 months and 6 months after treatment with CS. At each visit, all patients underwent symptom scoring and physical sign scoring. The level of eosinophil cationic protein (ECP) in the nasal discharge supernatants was measured by radioimmunoassay. The patients were divided into 2 groups according to nasal discharge EOS percentages, an EOS group (EOS > or = 3%) and a non-EOS group (EOS < 3%). The response to CS therapy (as measured by symptom and physical sign scores) and the changes of nasal discharge measurements were compared between the 2 groups. RESULT: In the EOS group, the baseline EOS [0.086 (0.065; 0.176)] and ECP level [(326 +/- 145) microg/L] were significantly higher than EOS [0.016 (0.005; 0.022)] and ECP level [(154 +/- 58) microg/L] of the non-EOS group, t = 4.40, 3.32, respectively, all P < 0.01. After 2 months and 6 months of CS therapy, the nasal discharge EOS, ECP level were 0.038 (0.006; 0.070), 0.019 (0.010; 0.060), (175 +/- 122) microg/L, (175 +/- 153) microg/L, respectively in the EOS group,which were significantly different as compared to baseline values (F = 6.73, 7.38, respectively all P < 0.05). But in the non-EOS group, the nasal discharge EOS and ECP level were 0.014 (0.004; 0.032), 0.015 (0.010; 0.026), (118 +/- 60) microg/L, (112 +/- 60) microg/L, respectively at 2 and 6 months, which showed that the nasal discharge EOS level and the symptom and physical sign scores did not improve significantly (F = 0.82, P > 0.05), but the ECP level did improve (F = 3.78, P < 0.05). The average daily dose of CS was not different between the two groups at any visits. CONCLUSION In persistent allergic rhinitis with low EOS in nasal discharge, CS therapy for 6 months failed to improve symptoms and physical signs.
Administration, Inhalation ; Adult ; Eosinophil Cationic Protein ; Eosinophils ; pathology ; Female ; Glucocorticoids ; administration & dosage ; Humans ; Leukocyte Count ; Male ; Middle Aged ; Rhinitis, Allergic ; drug therapy ; metabolism ; pathology ; Treatment Outcome

OBJECTIVE: To investigate the changes of leukotriene D4 (LTD4) in nasal discharge and plasma of patients with persistent allergic rhinitis (AR) and the effects of antihistamine. METHOD: The investigation was a prospective, randomized controlled trial. Forty AR patients (group C) were divided randomly into two subgroup. One group received oral antihistamine 10 mg everyday for one week (group CA) and another group received no loratadine tablets 10 mg everyday for one week (group CB). Fifteen age matched healthy (group D) people were enrolled as control. The level of LTD4 and interleukin-5 (IL-5) in both nasal discharge and plasma by using enzyme linked immunosorbent assay (ELISA) and enzyme immunoassay (EIA), cell counts and cell differentials in nasal discharge, were measured before and after three month. The clinical symptom and life quality scores of group C were also investigated. RESULT: The concentrations of LTD4 in nasal discharge [(794 +/- 305) pg] and plasma [(5219 +/- those in group D [(347 +/- 169) pg, (2283 +/- 489) ng/L, all P 1185) ng/L] in group C were significantly higher than those in group D [(347 +/- 169) pg, (2283 +/- 489) ng/L, all P < 0.05]. The level of LTD4 in nasal discharge was positively correlated with the percentage of neutrophil (r = 0.453, P < 0.05) and IL-5 (r = 0.364, P < 0.05). The pre- and post-therapy concentrations of nasal discharge and plasma in group CA were (812 +/- 1592) pg, (657 +/- 495) pg and (5422 +/- 935) ng/L, (4589 +/- 1057) ng/L respectively; While in group CB the concentrations were (776 +/- 227) pg, (860 +/- 194) pg and (5074 +/- 1850) ng/L, (6063 +/- 450) ng/L, respectively. There were no significant difference either in the level of LTD4 in nasal discharge or in plasma in both groups (all P > 0.05). CONCLUSION The results suggested that LTD4 was involved in airway inflammation in AR. Antihistamine was not effective enough in decreasing the levels of LTD4 in both nasal discharge and plasma of AR patients.
Adult ; Anti-Allergic Agents ; pharmacology ; Bodily Secretions ; chemistry ; Female ; Histamine H1 Antagonists ; pharmacology ; Humans ; Leukotriene Antagonists ; therapeutic use ; Leukotriene D4 ; analysis ; blood ; metabolism ; Male ; Middle Aged ; Plasma ; chemistry ; Prospective Studies ; Rhinitis, Allergic, Perennial ; blood ; drug therapy ; metabolism

OBJECTIVE: To assess the value of chromosomal karyotyping analysis and single nucleotide polymorphism-based microarray (SNP-array) for the detection of chromosomal mosaicisms in amniotic fluid samples. METHODS: Seventy four pregnant women with fetal mosaicisms detected by both methods were retrospectively analyzed. RESULTS: Among the 74 mosaicisms, 12 were pseudo and 62 were true mosaicisms, which included 1 Robertsonian translocation, 3 deletions, 4 supernumerary markers, 19 autosomal aneuploidy mosaicisms, 30 sex chromosome aneuploidy mosaicisms and 5 isometric chromosome mosaicisms. CONCLUSION Chromosome karyotyping analysis and SNP-array have their own advantages and limitations for the diagnosis of mosaicisms. When the two methods have yielded inconsistent results, fluorescence in situ hybridization may be used for further verification.
Aneuploidy ; Chromosome Aberrations ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Mosaicism ; Pregnancy ; Prenatal Diagnosis/methods* ; Retrospective Studies ; Sex Chromosome Aberrations

Objective:To evaluate the effect of botulinum neurotoxin A (BoNT/A) on prevention of chronic migraine (CM) in mice and explore the potential mechanism.Methods:Twenty-four male C57BL/6 mice were randomly divided into control group, nitroglycerin (NTG) group, and BoNT/A+NTG group ( n=8). Mice in the latter two groups were intraperitoneally injected with 10 mg/kg NTG on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments to establish CM models. Mice in the BoNT/A+NTG group were injected with 0.18 U/100 μL BoNT/A one h before the first injection of NTG. Mice in the control group were injected with the same dose of normal saline. Basal mechanical withdrawal threshold (MWT) and evoked MWT 2 h after NTG in the facial and hindpaw regions on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments were evaluated by von Frey filament test. The motor function of mice 2 h after NTG injection was tested by rotarod test on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments. On 9 th d of experiments, the mice were sacrified; the calcitonin gene-related peptide (CGRP), synaptosomal-associated protein 25 (SNAP25), glial fibrillary acidic protein (GFAP) and TRP channel protein expressions in the trigeminal ganglia (TG) and trigeminal nucleus caudalis (TNC), and NOD-like receptor protein 3 (NLRP3) inflammatory factor pathway-related protein expressions in TNC were detected by Western blotting; real-time quantitative PCR (RT-qPCR) was used to detect the NLRP3 inflammatory factor pathway-related mRNA expressions in TNC. The CGRP expression in TNC was detected by immunofluorescent staining. Results:(1) As compared with the control group, the NTG group had significantly decreased basal facial MWT on the 7 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased basal facial MWT on the 7 th and 9 th d of experiments ( P<0.05). As compared with the control group, the NTG group had significantly decreased evoked facial MWT on the 5 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased evoked facial MWT on the 5 th and 9 th d of experiments ( P<0.05). As compared with the control group, the NTG group had significantly decreased basal and evoked MWT in the hindpaw regions on the 3 rd, 5 th, 7 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased basal and evoked MWT in the hindpaw regions on the 3 rd, 5 th, 7 th and 9 th d of experiments ( P<0.05). (2) There was no significant difference in running time on rotarod among the three groups ( P>0.05). (3)Western blotting results showed that as compared with those in the control group, the CGRP and SNAP25 protein expressions were significantly increased in TG of the NTG group ( P<0.05); and those in the BoNT/A+NTG group were significantly decreased as compared with those in the NTG group ( P<0.05). As compared with those in the control group, the CGRP and NLRP3 protein expressions were significantly increased in TNC of NTG group ( P<0.05); and those in the BoNT/A+NTG group were significantly decreased as compared with those in the NTG group ( P<0.05). (4)RT-qPCR results showed that as compared with that in the control group, the IL-1β mRNA expression in TNC of the NTG group was significantly increased ( P<0.05), and that in the BoNT/A prevention group was statistically decreased as compared with that in the NTG group ( P<0.05). (5) Immunofluorescent staining results showed that as compared with that in the control group, the CGRP expression in TNC of the NTG group was significantly increased, and that in the BoNT/A+NTG group was significantly decreased as compared with that in the NTG group ( P<0.05). Conclusion:BoNT/A can reduce the SNAP25 expression in TG, reduce the CGRP release in TG and TNC, and prevent CM onset; BoNT/A can regulate NLRP3 level in TNC.

Objective This project aims to establish an interactive oral and maxillofacial radiological image annotation database and to analyze its feasibility for implementation into curricula in order to develop a highly effective software for image browsing. Methods We established the interactive image annotation database primarily on the basis of the local net-work and Foxit Reader. A pilot survey was then conducted to determine the performance of the interactive database. Seventy-six students were asked to complete a structured and open questionnaire related to their perceptions of using the database. Simple numeric quantitative and qualitative analyses were then applied. Results A total of 542 portable document format (PDF) digital teaching images and corresponding annotated files were collected. The survey revealed that most of the students found the database useful. Approximately 87.50% of the 64 subjects who compelete questionnaire believed that the database was superior to an older system (joint photographic experts group, JPEG) of image browsing. Conclusion The integration and sharing of teaching resources and the establishment of an internet-based learning platform is the key to realizing a digital medical teaching system. The established database has high potential in a wide range of practical applications.