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ObjectiveTo explore the possible peripheral analgesic mechanism of electroacupuncture （EA） at promimal and distal acupoints in treatment of myofascial pain syndrome （MPS）. MethodsTwenty-four SD rats were randomly divided into blank group， model group， proximal group， and distal group， with six rats in each group. MPS model was prepared by “strike combined with centrifugal exercise” in all groups except for the blank group. After modeling， the rats in the proximal group received EA at the local myofascial trigger points （MTrPs）， namely the Ashi points， with dilatational waves of frequency of 2/100 HZ and voltage of 2-4 V， current intensity depending on a slight trembling of the left lower limbs， once a day， 15min each time，for 14 days. The rats in the distal group received EA at “Yanglingquan” （GB 34） and “Yinlingquan” （SP 9）， with the same operations as the proximal group. The rats in the blank group and the model group were only grasped and hedged， without other interventions. After intervention， the paw withdrawl mechanical threshold （PWMT） was measured， and variability between the left and right hind paws was calculated. Musculoskeletal ultrasound imaging and electromyography monitoring were performed on the left lower extremity vastus medialis. The morphological changes of vastus medialis muscle of the left lower extremity were observed by HE staining. The positive expression of substance P （SP）， calcitonin gene-related peptide （CGRP）， CD68 and CD206 in muscle tissue was detected by immunohistochemistry. Abdominal aortic serum interleukin-1β （IL-1β）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interleukin-10 （IL-10）， and interleukin-8 （interleukin-8） were detected by ELISA. ResultsCompared to those in the blank group， the fibers of the vastus medial muscle of the rats in the model group were broken and distorted with thickness in variation， and the myofascia was broken， with fibrillation potential， enlarged muscle cells， inward moved nucleus， and widened muscle space； the variability of PWMT between the left and right hind paws significantly increased， as well as the levels of SP， CGRP， CD68， and CD206 in the vastus medialis muscle （P＜0.01）， and the serum IL-8 and TNF-αlevels were significantly elevated （P＜0.05 or P＜0.01）. Compared to those in the model group， the muscle fibers in the proximal and distal group were complete in shape and arranged in an orderly manner， with continued non-broken myofascia， regular shape of muscle cells， and significantly reduced level of IL-8 （P＜0.01）； the amplitude and frequency of spontaneous discharge in the proximal group significantly decreased， as well as the variability of PWMT between the left and right hind paws， and the levels of SP， CGRP， and CD68 in the vastus medialis muscle， while the CD206 level increased significantly （P＜0.05 or P＜0.01 ）； there was complex discharges in the distal group， with significantly decreased level of CD68 in the vastus medialis muscle and increased level of CD206 （P＜0.01）. Compared to the proximal group， the level of IL-8 in the distal group was significantly higher （P＜0.05）. ConclusionsEA at proximal acupoints can significantly improve the pain threshold and local muscle tissue morpho-logy in rats， and its mechanism may be related to reducing the levels of pain-causing substances and related inflammatory factors and promoting the polarization of macrophages. The analgesic effect of EA at distal acupoints is not obvious， and the mechanism is still unclear.
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Objective To establish a solvent desorption inductively coupled plasma-mass spectrometry (ICP-MS) method for determination of iodine in workplace air. Methods Iodine in workplace air was collected with alkaline activated carbon tube and desorbed with 10.0 mL pure water or 20 mmol/L sodium bicarbonate solution. Rhenium-185 was used as an internal standard for quantification. The sample was determined in standard mode and kinetic energy discrimination collision (KED) mode by ICP-MS. Results In standard mode, iodine showed a good linear range in the concentration of 9.0 to 1 100.0 μg/L, with a correlation coefficient of 0.999 3 and a detection limit of 2.7 μg/L. In KED mode, iodine showed a good linear range in the concentration of 24.3 to 800.0 μg/L, with a correlation coefficient of 0.999 1 and a detection limit of 7.3 μg/L. The average desorption efficiency using pure water ranged from 99.1% to 106.7%, with within-run relative standard deviation (RSD) of 3.1% to 8.0% and between-run RSD of 4.9% to 9.3%. The average desorption efficiency using sodium bicarbonate solution ranged from 96.5% to 105.3%, with within-run RSD of 4.9% to 8.6% and between-run RSD of 2.5% to 9.9%. There were no statistical significant differences in the main effects of desorption solution, ICP-MS detection mode, their interaction on average desorption efficiency and within-run RSD (all P>0.05). Samples could be stored at room temperature for at least 7 days. Conclusion This method is highly sensitive, accurate, and suitable for the determination of iodine in workplace air. The sample pretreatment is simple and rapid.
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OBJECTIVE To provide reference for improving the professional identity of clinical pharmacists and the quality of pharmaceutical care ，and promoting the effects of clinical pharmaceutical intervention. METHODS A questionnaire survey was conducted among clinical pharmacists in secondary and tertiary hospitals in 31 provinces（autonomous regions and municipalities ） in 2019 by stratified semi-random sampling. Through descriptive analysis of survey data ，their job satisfaction status was evaluated ； χ 2 test and Logistic regression analysis were used to analyze the influential factors of job satisfaction ；the robustness test of study results by propensity score matching method and replacement regression model ，and grouping Logistic regression of samples from hospital on different levels. Targeted improvement measures were put forward according to the results of survey . RESULTS There was statistical significance in the difference of job satisfaction among pharmacists of different professional titles （P＜0.05）. Results of Logistic regression showed that whether to participate in standardized training ，whether to obtain communication and support from patients ，whether the pharmaceutical management rules and regulations were sound ，whether to set up economic compensation means such as pharmaceutical service fee ，whether to work overload ，and whether to smoothly perform pharmaceutical care duties were significant influential factors for job satisfaction of clinical pharmacists （P＜0.05）. These results showed good robustness as tested by propensity score matching method and replacement regression model. Heterogeneity analysis results showed that the job satisfaction of clinical pharmacists in tertiary hospitals was more significantly affected by economic compensation ，while clinical pharmacists in secondary hospitals were more concerned about training opportunities and workload conditions （P＜0.05）. CONCLUSIONS The job satisfaction level of Chinese clinical pharmacists remains to be improved. Accordingly ，it is compulsory to continue the promotion of standardized training courses ，consummate the pharmaceutical management system ，and fair remuneration structure in order to improve the job satisfaction of clinical pharmacists and build a high-level clinical pharmacist team.
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Objective:To express virus-like particles of poliovirus type 2 (PV2-VLP) in insect cells using a recombinant baculovirus expressing P1 and 3CD and to preliminarily evaluate its immunogenicity.Methods:Based on the codon preference of High 5 cells, the sequences of P1 gene and 3CD gene of PV2 were optimized and inserted into pUC57-Amp to construct pUC57-PV2-P1 and pUC57-PV2-3CD. UC57-PV2-P1s mutant that carried P1 gene mutation affecting thermostability was then constructed. Recombinant baculovirus strains of rBac-PV2-P1s-3CD and rBac-PV2-P1-3CD (wild type) were constructed using homologous recombination. The expression of target proteins was detected by Western blot. PV2-VLP was purified by ion exchange chromatography. The structure of VLP was observed under transmission electron microscopy to evaluate the assembly efficiency. The immunogenicity of PV2-VLP was assessed in a rat model.Results:The recombinant baculovirus with stable expression of P1s and 3CD proteins was successfully constructed. Western blot results showed that the yield of VLP was higher after thermostability mutation than that of the wild type. A three-dimensional structure with a diameter of about 30 nm was observed under electron microscopy, indicating that the VLP was successfully assembled. Animal experiment showed that the recombinant PV2-VLP had immunogenicity and could effectively induce the production of neutralizing antibodies.Conclusions:Effective VLP vaccines could be successfully prepared using the insect cell-baculovirus expression system, which provided reference for the development of polio VLP vaccine.
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Objective:To screen the neutralizing epitope of enterovirus 71 (EV71) and determine the specific minimum amino acid sequence that triggers immunity for providing a theoretical basis for the development of synthetic peptide vaccines.Methods:EV71 neutralizing antibody-specific binding clones were panned and sequenced using a phage display random 12-peptide library to obtain the key sequences of neutralizing epitopes. A series of peptides containing the key sequences with N-terminal acetylation (AC) and C-terminal linking to Keyhole limpet hemocyanin (KLH) were synthesized. Serum samples were collected after immunizing mice with the modified peptides. Then the immunogenicity of the peptides and the neutralizing activity of serum samples were analyzed by Western blot, ELISA and neutralization test.Results:After three rounds of panning, cloning and sequencing, KQEKDL was identified as the key motif. The serum samples collected from the mice immunized with the modified series of peptides containing key motifs had different degrees of binding ability to EV71 and VP1 protein. The serum samples of mice immunized the synthetic peptide containing only the minimum key motif (AC-KQEKDL-KLH) had the strongest response to the other three peptides and EV71 and the highest neutralizing titer.Conclusions:The EV71 neutralizing epitope was successfully screened using the phage display random peptide library. The key motif of KQEKDL might be the specific minimum amino acid sequence that triggered the immune system. This study provides a theoretical basis for better understanding the immune response mechanism, evaluating the immunogenicity of the antigens and further research and development of polypeptide vaccines.
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Objective:To investigate the humoral immune response to GⅠ.1/GⅡ.4 norovirus (NoV) virus-like particle (VLP) vaccine of different doses in mice.Methods:The GⅠ.1/GⅡ.4 norovirus vaccine was diluted into four different concentrations, containing 50, 25, 8.3 and 2.8 μg/dose antigens, respectively. Aluminum hydroxide adjuvant was used in the control group. Ten 7-week-old BALB/c mice in each group were immunized intraperitoneally with 0.5 ml vaccine or adjuvant on 0 d and 21 d. Blood samples were collected on 35 d and the histoblood group antigen (HBGA) blocking titers of GⅠ.1 and GⅡ.4 antibodies in serum were detected. The differences in antibody levels between the groups were analyzed by SPSS23.0 software.Results:GⅠ.1 and GⅡ.4 HBGA blocking antibodies in 50, 25 and 8.3 μg/dose groups became positive on 35 d after the first dose vaccination. The geometric mean titers (GMT) of GⅠ.1 and GⅡ.4 HBGA blocking antibodies were 488 (95%CI: 249-955) and 489 (95%CI: 302-790) in 50 μg/dose group, 278 (95%CI: 106-728) and 738 (95%CI: 299-1 820) in 25 μg/dose group, 300 (95%CI: 158-570) and 486 (95%CI: 222-1 068) in 8.3 μg/dose group, respectively. The positive rates of GⅠ.1 and GⅡ.4 blocking antibodies in 2.8 μg/dose group were 40% and 70% and the GMT were 23 (95%CI: 10-51) and 85 (95%CI: 24-304), respectively. The GⅠ.1 and GⅡ.4 blocking antibodies in the control group were negative. Statistically significant differences in antibody levels were found between 50, 25, 8.3 μg/dose groups and the control group ( P<0.05), as well as in GⅠ.1 blocking antibodies between 50, 25, 8.3 μg/dose groups and 2.8 μg/dose group ( P<0.05). GⅡ.4 antibody level in 25 μg/dose group was statistically different from that in 2.8 μg/dose group ( P<0.05). Conclusions:GⅠ.1/GⅡ.4 norovirus VLP vaccine at (50-8.3) μg/dose could induce humoral immune response in mice.
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Objective@#Longitudinal data were used in this study to examine the predictive effects of psychological stress in early puberty on subsequent anxiety and depression symptoms.@*Methods@#Objects from a puberty cohort of 998 children from 4 primary schools in Chongqing were included. Psychological stress was measured during their early puberty, and anxiety and depression were followed up after 4 years. Multiple linear regression was used to analyze the influence of psychological stress on anxiety and depression level in their middle puberty.@*Results@#The average score of psychological stress during early puberty was (34.79±24.78), and scores of anxiety and depression were (11.20±10.89) and (12.06±6.69), respectively, with detection rates of 14.03% and 15.63%. Girls had higher anxiety and depression scores than boys(F=51.58,5.48，P<0.05). The depression scores of children with different parents’ educational levelsand perceived parental relationship were different(F=6.74, 7.38, 10.49, P<0.05). The results of multiple linear regression showed that girls(β=4.38), higher psychological stress(β=0.13), older age (β=0.89) were risk factors for higher anxiety level. Children with higher psychological stress(β=0.05), older age(β=0.57), perceived worse parents’ relationship(β=1.19), lower maternal education (β=-1.00) had higher depression scores(P<0.05).@*Conclusion@#The psychological stress level, and age during early puberty had a positive predictive effect on anxiety and depression after 4 years. Simultaneously, girls were more prone to anxiety, and poor parental relationship and low maternal literacy were risk factors for children’s depression.
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Objective To validate a cell infection-based quantitative RT-PCR for evaluating the potency of rotavirus vaccine. Methods According to the ICH ( the International Council for Harmonization) Harmonised Tripartite Guideline, the method was validated for its specificity, accuracy, precision, linearity and robustness. Results The method had good specificity as it could only amplify and detect the corre-sponding type of rotavirus strain. The recovery rates for determining the potency against rotaviruses of G2, G3 and G4 types were 97％ to 108％. The percent coefficient of variation ( CV) of both intra-plate and in-ter-plate precision was≤2. 62％, while the intraday and interday CV was≤1. 76％ and≤2. 27％, respec-tively. The CV between the two experimenters was≤7. 68％. The linearity range of the method was 4. 4-6. 5 UI for G2 type rotavirus, 3. 9-8. 3 UI for G3 type and 3. 5-8. 1 UI for G4 type. Good robustness was observed using the cells of 140 to 160 generations. Conclusions The cell infection-based quantitative RT-PCR was shown to have satisfactory specificity, accuracy, precision, linearity and robustness, suggesting that it was a suitable method for evaluating the potency of multivalent rotavirus live vaccines.
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Objective To evaluate whether simultaneous vaccination with live attenuated polio vaccine affects the immunogenicity of live attenuated rotavirus ( RV) vaccine. Methods Rotarix produced by GlaxoSmithKline was used as the research object. Two doses of Rotarix were orally administered on day 0 and month 1, and oral live attenuated polio vaccine (OPV) was administered on day 0, month 1 and month 2 according to the national vaccination plan. Healthy infants aged 6 to 16 weeks were randomly divided into two groups:interval vaccination group ( Rotarix and OPV were vaccinated on different days) and simultane-ous vaccination group ( Rotarix and OPV were vaccinated on the same day) . Serum samples were collected on day 0, month 2 and month 12, and serum RV-IgA was measured by enzyme linked immunosorbent assay. Statistical analysis was performed to evaluate whether there were statistical differences in the seroconversion rate and level distribution of RV-IgA between the two groups. Results The seroconversion rate of serum RV-IgA in month 2 was 73. 84％ in the interval vaccination and 63. 95％ in the simultaneous vaccination group, and the difference between them was statistically significant (P<0. 05). The geometric mean concen-trations (GMC) of RV-IgA were 97 EU/ml and 90 EU/ml, respectively (P>0. 05). Compared with the simultaneous vaccination group, the seroconversion rate and GMC of serum RV-IgA in month 12 were higher in the interval vaccination group, and the differences were statistically significant (P<0. 05). Conclusions Simultaneous vaccination with live attenuated polio vaccine would affect the immune response of live attenua-ted rotavirus vaccine, especially the maintenance of RV-IgA antibody level.
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Objective@#To evaluate whether simultaneous vaccination with live attenuated polio vaccine affects the immunogenicity of live attenuated rotavirus (RV) vaccine.@*Methods@#Rotarix produced by GlaxoSmithKline was used as the research object. Two doses of Rotarix were orally administered on day 0 and month 1, and oral live attenuated polio vaccine (OPV) was administered on day 0, month 1 and month 2 according to the national vaccination plan. Healthy infants aged 6 to 16 weeks were randomly divided into two groups: interval vaccination group (Rotarix and OPV were vaccinated on different days) and simultaneous vaccination group (Rotarix and OPV were vaccinated on the same day). Serum samples were collected on day 0, month 2 and month 12, and serum RV-IgA was measured by enzyme linked immunosorbent assay. Statistical analysis was performed to evaluate whether there were statistical differences in the seroconversion rate and level distribution of RV-IgA between the two groups.@*Results@#The seroconversion rate of serum RV-IgA in month 2 was 73.84% in the interval vaccination and 63.95% in the simultaneous vaccination group, and the difference between them was statistically significant (P<0.05). The geometric mean concentrations (GMC) of RV-IgA were 97 EU/ml and 90 EU/ml, respectively (P>0.05). Compared with the simultaneous vaccination group, the seroconversion rate and GMC of serum RV-IgA in month 12 were higher in the interval vaccination group, and the differences were statistically significant (P<0.05).@*Conclusions@#Simultaneous vaccination with live attenuated polio vaccine would affect the immune response of live attenuated rotavirus vaccine, especially the maintenance of RV-IgA antibody level.
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Objective To compare the relationship between the enzyme‐linked immunosorbent assay(ELISA) reagent and West‐ern blot(WB) confirmation reagent for analyzing the quality lever of human T‐cell lymphotropic virus(HTLV) detection reagent . Methods The WB confirmation reagent was used to detect anti‐HTLV antibody in 156 human serum samples of ELISA prelimina‐ry screening positive .The ELISA cut‐off value(optimal value) was selected by using the two‐graph receiver operating characteristics (TG‐ROC) analytical method .The two‐by‐two table analysis was constructed to analyze the consistency of results detected by the two methods ,moreover the McNemar test was used to evaluate the consistency of detection results .The quality level of HTLV de‐tection reagent was comprehensively evaluated .Results Among 156 serum samples of ELISA preliminary screening positive ,only 40 samples were positive by the WB confirmation ,and other 116 samples were negative .The sensitivity and specificity of ELISA de‐tection reagent obtained by TG‐ROC analysis were 97 .5% and 45 .7% respectively ,the TG‐ROC test also indicated that the detec‐tion results had significant difference between ELISA and WB(P<0 .05) .By adjusting the cut‐off value ,the sensitivity and specific‐ity of ELISA were increased to 88 .8% (parametric method) .In the comparison of the parametric method and the non‐parametric method ,the obtained areas under the curve(AUC) was 0 .923 5(parametric method) ,their results were basically consistent .Conclu‐sion Although above results indicate that the detection results of ELISA reagent are different from those of WB ,but adjusting the cut off value can increase its sensitivity and specificity ,thus increases the reliability of diagnosis result .
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Objective To establish a culture system in vitro of adult human retinal neural cells for providing a model for the research of retinal neural cells. Design Experimental study. Participants Cultured adult human retinal neural cells. Methods The isolated cells from adult human postmortem retina (20?40 years old) were cultured, then cells of different stages were identified with immunocytochemical staining and judged with phase contrast microscopy and electron microscopy. Main Outcome Measures Cellular morphology and structure. Results (1) The results of cell culture: the adult retinal neural cells could survive in vitro under some conditions and were identified as NSE positive mostly. (2) The results of electron microscopy: most cultured cells were photoreceptors, bipolar cells, horizontal cells and some were glial cells with scanning electron microscopy. Conclusions Under feasible conditions, the adult human retinal neural cells could be cultured and maintained effectively in vitro.
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Objective To explore the effects of drugs on functions of mitochondria in retinal nerve cells, and to lay a foundation of the investigation of drug protection for retinal nerve cells. Methods Cultivation of the retinal nerve cells of 8 eyes of neonatal calves was performed. The changes of fluorescent density of the mitochondria of cultured cells labeled by dye rhodamine 123 (Rh123) before and after the activation of the medicines, including ferulic acid (FA), arginine, glycine, taurine, vitamine E and brain-derived neurotrophic factor (BDNF) respectively, were detected by laser-scanning confocal microscopy. Results FA with the concentration of 500 ?g/ml led the diphasic variation of the fluorescent intensity of mitochondria. After scanning for 60.772 seconds when treated with FA firstly, the fluorescent intensity decreased rapidly (from 45.425?4.153 to 22.135?5.293); while after 112.774 seconds when treated secondly, the intensity increased obviously (from 19.655?4.383 to 28.247?4.764), and after 168.773 seconds when treated thirdly the intensity still increased. After scanning for 56.457 seconds when treated with vitamin E (12.5 mg/ml), the fluorescent intensity increased obviously (from 88.255?5.039 to 111.273?4.529), which suggested that vitamin E with the concentration of 12.5 mg/ml strengthen the fluorescent intensity. After scanning for 58.147 and 134.148 seconds when treated with BDNF (50 ng/ml) respectively, the fluorescent intensity increased obviously (from 69.115?5.038 to 77.225?5.131) which suggested that BDNF with the concentration of 50 ng/ml led the increase of the fluorescent intensity. Glycine (2.5 mg/ml) and arginine(30 mg/ml) didn't affect the fluorescent intensity of mitochondria, and taurine (6.25 mg/ml) caused the appreciable decrease of the fluorescent intensity. Conclusion FA, BDNF and vitamin E may promote the metabolism of retinal nerve cells via the path of mitochondria, while amino acids may adjust the activation of retinal nerve cells through other ways.