Objective To explore the effect and preliminary mechanism of the plant-derived immunostimulatory molecule,ophiopogonin,on enhancing the immune response of a subunit vaccine with the receptor-binding domain(RBD)of coronavirus spike protein as the antigen.Methods CCK-8 assay was used to determine the cytotoxicity of ophiopogonin D'(OPD')on bone marrow-derived dendritic cells(BMDCs).Female Balb/c mice were randomly divided into RBD,RBD/OPD',RBD/Alum,and control groups.The immunization dose was 5 μg of antigen per mouse and 100 μg of adjuvant per mouse,and immunization was carried out according to the intramuscular injection immunization procedure on days 0,21,and 42.The titers of specific IgG and its subtype antibodies were detected by ELISA.The cytokine levels in the supernatant of splenocytes were detected using ELISA.The number of splenocytes secreting IFN-γ was detected by ELISpot.Laser confocal microscopy was employed to observe the uptake of antigen by BMDCs.The phagocytic ability of BMDCs for antigen was quantitatively analyzed by flow cytometry.The mechanism of its enhanced immune effect was preliminarily explored using transcriptomics technology combined with bioinformatics research.Results When the concentration of OPD'was less than 5 μg/mL,the survival rate of BMDCs was 100%.After a single intramuscular injection in mice,except for a slight decrease in body weight,the other biochemical indicators were within corresponding normal ranges.After intramuscular injection immunization of the vaccine,the titers of serum-specific IgG,IgG1,and IgG2a in the RBD/OPD'group were significantly higher than those in the RBD group(P<0.05).Compared with the RBD group,the RBD/OPD'group induced a high-level Th1 cell immune response of IL-1β,TNF-α,and IFN-γ(P<0.01)and had more lymphocytes secreting IFN-γ(P<0.001).Laser confocal microscopy displayed that BMDCs took up more antigens after OPD'treatment,which was further confirmed with flow cytometry in quantitative analysis on antigen uptake rate(P<0.01).Transcriptomics results indicated that there was more significant enrichment of the PPAR signaling pathway in the RBD/OPD'group than the RBD group,suggesting that OPD'may activate the PPAR signaling pathway to exert its adjuvant effect.Conclusion OPD'effectively enhances the immune response of the RBD subunit vaccine,and its action mechanism may be related to the activation of the PPAR signaling pathway.

Objective To prepare tubeimoside Ⅲ nanoemulsion(TBMⅢ-NE)and evaluate its adjuvant effect in vaccines.Methods TBMⅢ-NE was prepared using low-energy emulsification.Dynamic light scattering was used to characterize the particle size and polydispersity index of the obtained TBMⅢ-NE,and transmission electron microscopy(TEM)was employed to observe the morphology.CCK-8 assay was utilized to determine the cytotoxicity of TBMⅢ-NE on bone marrow-derived dendritic cells(BMDCs).The in vitro safety of TBMⅢ-NE was evaluated using a hemolysis assay.The ability of TBMⅢ-NE to promote the phagocytosis of antigens by DC2.4 cells was observed using confocal laser microscopy.After co-incubation of TBMⅢ-NE with BMDCs,the expression levels of CD40,CD86,MHC-Ⅰ,and CCR7 on the surface of BMDCs were detected using flow cytometry,and the levels of cytokines in the supernatant of BMDCs were measured using enzyme-linked immunosorbent assay(ELISA).After female BALB/c mice were immunized with the SARS-CoV-2 antigen RBD in combination with TBMⅢ-NE,ELISA was conducted to determine the serum levels of specific IgG,IgG2a,and IgG1 antibodies.The number of specific IFN-γ-secreting cells in mouse splenocytes was detected using enzyme-linked immunospot(ELISpot)assay.Results The prepared blank nanoemulsion(BNE)and TBMⅢ-NE were in a particle size of 25.46 and 25.89 nm,and a polydispersity index of 0.214 and 0.125,respectively.TEM displayed that TBMⅢ-NE was in uniform sphere and well dispersed.When the TBMⅢ-NE adjuvant was diluted by 400-fold,the survival rate of BMDCs was approximately 86%.Compared with free TBMⅢ,the hemolytic toxicity of TBMⅢ-NE was significantly reduced(P<0.01).TBMⅢ-NE promoted the phagocytosis of antigens by DC2.4 cells and significantly increased the expression of CCR7 on the surface of BMDCs(P<0.05),indicating its potential to promote more dendritic cells to effectively migrate to lymph nodes.TBMⅢ-NE also promoted the expression of IL-6 and IL-1β in the supernatant of BMDCs(P<0.05).When combined with RBD,TBMⅢ-NE significantly increased the levels of specific IgG,IgG2a,and IgG1 antibodies in mouse serum(P<0.01)and promoted the secretion of specific IFN-γ in splenocytes(P<0.01),indicating that TBM Ⅲ-NE could enhance specific cellular immune responses.Conclusion A stable and highly effective TBMⅢ-NE that can induce humoral and cellular immune responses is successfully prepared.

OBJECTIVE To establish a extraction process of Changyan Heji Ⅱ(CYHJ-Ⅱ)based on UPLC-Q-TOF-MS/MS technology combined with response surface analysis,and to optimize the extraction process.METHODS The chemical components in CYHJ-Ⅱ were qualitatively analyzed by UPLC-Q-TOF-MS/MS technology,and the chemical components with good linear relation-ship in mass spectrometry response were selected as process investigation indicators;the extraction process parameters(water addition amount,extraction time and soaking time)were investigated by Box-Behnken design;the comprehensive score was obtained by princi-pal component analysis(PCA),and the optimal process was determined by the comprehensive score combined with response surface a-nalysis.RESULTS Through qualitative analysis,110 components were inferred and identified from CYHJ-Ⅱ,including 2 organic acids,82 flavonoids,13 terpenoids,and 13 alkaloids.Based on the results of qualitative analysis,48 index components with good lin-ear relationships were derived by UPLC-Q-TOF-MS/MS combined with Masshunter mass spectrometry data analysis software.PCA was performed and the comprehensive score was calculated.Response surface analysis was performed with the comprehensive score as an indicator.The optimal extraction process obtained by combining the response surface prediction results and actual production was:soaking for 45 min,8 times the amount of solvent,2 extractions,each time for 120 min.CONCLUSION This study provides a new idea for the investigation of the extraction process of traditional Chinese medicine compound prescriptions and expands a new method for the development of traditional Chinese medicine compound prescriptions.

OBJECTIVE To establish a extraction process of Changyan Heji Ⅱ(CYHJ-Ⅱ)based on UPLC-Q-TOF-MS/MS technology combined with response surface analysis,and to optimize the extraction process.METHODS The chemical components in CYHJ-Ⅱ were qualitatively analyzed by UPLC-Q-TOF-MS/MS technology,and the chemical components with good linear relation-ship in mass spectrometry response were selected as process investigation indicators;the extraction process parameters(water addition amount,extraction time and soaking time)were investigated by Box-Behnken design;the comprehensive score was obtained by princi-pal component analysis(PCA),and the optimal process was determined by the comprehensive score combined with response surface a-nalysis.RESULTS Through qualitative analysis,110 components were inferred and identified from CYHJ-Ⅱ,including 2 organic acids,82 flavonoids,13 terpenoids,and 13 alkaloids.Based on the results of qualitative analysis,48 index components with good lin-ear relationships were derived by UPLC-Q-TOF-MS/MS combined with Masshunter mass spectrometry data analysis software.PCA was performed and the comprehensive score was calculated.Response surface analysis was performed with the comprehensive score as an indicator.The optimal extraction process obtained by combining the response surface prediction results and actual production was:soaking for 45 min,8 times the amount of solvent,2 extractions,each time for 120 min.CONCLUSION This study provides a new idea for the investigation of the extraction process of traditional Chinese medicine compound prescriptions and expands a new method for the development of traditional Chinese medicine compound prescriptions.

ObjectiveTo investigate the measurement invariance and the score distribution characters of the 9-item Patient Health Questionnaire （PHQ-9）， and to test its feasibility in comparing depression among adolescents of different sexes and ages. MethodsFrom September 2018 to May 2019， 12 241 adolescents aged 10 to 19 years old across 22 primary and secondary schools in Hangzhou were included based on cluster sampling method， and they were investigated by PHQ-9. The measurement invariance was tested by multi-group confirmatory factor analysis. Poisson regression analysis and Logistic regression analysis were adopted to explore the score distribution of PHQ-9 by sex and age. ResultsThe configural， metric， scalar and strict invariances （χ²=2 492.527， df=79， P<0.01， RMSEA=0.071， SRMR=0.031， CFI=0.942， TLI=0.947， ΔCFI=0.004， ΔTLI=0.002 compared with the scalar model） of PHQ-9 across sex were all acceptable， and the configural， metric and scalar invariances （χ²=3 311.991， df=414， P<0.01， RMSEA=0.076， SRMR=0.055， CFI=0.928， TLI=0.937， ΔCFI=0.010， ΔTLI=0.002 compared with the metric model） across age were acceptable. The detection rate of mild depressive symptoms in girls was higher than that in boys （52.99% vs. 46.65%， χ²=48.344， P<0.01）. The PHQ-9 total scores of girls aged 13 to 17 were higher than those of boys （D=0.092~0.144， P<0.01）. There was an interaction between sex and age （χ²=32.800， df=1， P<0.01， OR=0.895）. Except low self-evaluation and suicide or self-injury， the detection rate of assessed symptom on each item of girls was higher than that of boys （P<0.05）. ConclusionIt is discovered that PHQ-9 has measurement equivalence across sex and age in adolescents， and girls and older adolescents scored higher in PHQ-9.

Objective:To analyze the monitoring results of iodine deficiency disorders in Nanning City in 2020, learn about the consumption of iodized salt among residents and the iodine nutrition status of key populations, and provide scientific basis for formulating or adjusting targeted prevention and control measures of iodine deficiency disorders.Methods:According to the Guangxi Iodine Deficiency Disorders Monitoring Plan, monitoring was carried out in all of 12 districts (counties) in Nanning City. One township (street) was selected from each of the five directions of east, west, south, north, and central. Forty non-boarding children aged 8 to 10 and 20 pregnant women were selected as monitoring subjects in each township (street). Edible salt samples were collected from children and pregnant women to detect salt iodine content, random mid-course urine samples from children and morning urine samples from pregnant women were collected to detect urinary iodine content; in addition, thyroid examination of children was conducted in Qingxiu District, Liangqing District, Long an County and Shanglin County.Results:In 2020, a total of 2 434 children aged 8 to 10 and 1 207 pregnant women were surveyed in Nanning City. The coverage rate of iodized salt, the qualified rate of iodized salt and the qualified iodine salt consumption rate were 99.67%(3 629/3 641), 97.99%(3 556/3 629) and 97.67%(3 556/3 641), respectively. The median urinary iodine of children was 182.0 μg/L, and the median values ranged from 146.5 to 234.8 μg/L in different districts (counties), there were significant differences in median urinary iodine between urban and non-urban areas, different gender and age groups ( U = 2.38, 2.41, P = 0.017, 0.016; H = 16.42, P < 0.001). The goiter rate of children was 0.99%(8/807), and the rate ranged from 0 to 2.00% (4/200) in the 4 districts (counties) examined, there were significant differences in thyroid volume between urban and non-urban areas and different ages ( U = - 3.52, P < 0.001; H = 47.67, P < 0.001). The median urinary iodine of pregnant women was 191.0 μg/L. The median urinary iodine of different districts (counties) ranged from 141.0 to 241.5 μg/L, and the difference was statistically significant at different gestational stages ( H = 24.37, P < 0.001). Conclusion:In 2020, the coverage rate of iodized salt, the qualified rate of iodized salt and the qualified iodine salt consumption rate by residents in Nanning City are high, and the iodine nutrition of both children and pregnant women are at an appropriate level.

As a ligand-dependent transcription factor,retinoid-associated orphan receptor γt(RORyt)that controls T helper(Th)17 cell differentiation and interleukin(IL)-17 expression plays a critical role in the pro-gression of several inflammatory and autoimmune conditions.An emerging novel approach to the therapy of these diseases thus involves controlling the transcriptional capacity of RORyt to decrease Th17 cell development and IL-17 production.Several RORyt inhibitors including both antagonists and inverse agonists have been discovered to regulate the transcriptional activity of RORyt by binding to orthosteric-or allosteric-binding sites in the ligand-binding domain.Some of small-molecule inhibitors have entered clinical evaluations.Therefore,in current review,the role of RORyt in Th17 regulation and Th17-related inflammatory and autoimmune diseases was highlighted.Notably,the recently developed RORyt inhibitors were summarized,with an emphasis on their optimization from lead compounds,ef-ficacy,toxicity,mechanisms of action,and clinical trials.The limitations of current development in this area were also discussed to facilitate future research.

Ulcerative colitis (UC) is one of types of inflammatory bowel disease with high recurrence. Recent studies have highlighted that microbial dysbiosis as well as abnormal gut immunity are crucial factors that initiate a series of inflammatory responses in the UC. Modulating the gut microbiota-intestinal immunity loop has been suggested as one of key strategies for relieving UC. Many Chinese herbal medicines including some of single herb, herbal formulas and the derived constituents have been reported with protective effect against UC through modulating gut microbiome and intestinal immunity. Some clinical trials have shown promising results. This review thus focused on the current knowledge on using Chinese herbal medicines for treating UC from the mechanism aspects of regulating intestinal homeostasis involving microbiota and gut immunity. The existing clinical trials are also summarized.

Objective:To survey the awareness status of risk factors for gastric cancer and related factors among residents in Shijiazhuang city.Methods:From October to November 2020, residents aged>18 years were selected through convenience sampling from Xinhua District and Yuhua District in Shijiazhuang for a face-to-face questionnaire survey. The demographic characteristics and awareness levels of risk factors for gastric cancer among 1 490 subjects were analyzed. The multivariate linear regression model was applied to analyze the related factors.Results:There were 522 male participants (35.0%) and 968 female participants (65.0%). The mean knowledge score of risk factors of gastric carcinoma in the respondents was 5.0 (1.0, 11.0). There were 57.0% (849/1 490) of the participants who had a low knowledge level about risk factors of gastric cancer. The more highly recognized risk factors were irregular diet (47.4%, 706/1 490), consumption of pickled foods (45.2%, 674/1 490), consumption of smoked foods (45.0%, 671/1 490); the less-recognized risk factors included physical inactivity (14.8%, 221/1 490), male sex (17.3%, 258/1 490) and older age (19.5%, 291/1 490). Only 26.8% (400/1490) of the participants regarded H. pylori infection as a risk factor of gastric cancer. Univariate analysis showed that educational level ( Z=39.34), marital status ( Z=53.31), monthly income ( Z=11.82), family member or friend ever having stomach problem ( H=-2.98), and family history of gastric cancer ( H=-2.34) were significantly associated with the knowledge score of risk factors for gastric cancer ( P<0.05). Multivariate analysis showed that compared to participants with educational levels of primary school or below, those with educational level of junior high school ( β=0.27, P<0.001), high school ( β=0.23, P<0.001), or college ( β=0.16, P<0.001) had a higher levels of knowledge of risk factors for gastric cancer; compared to unmarried participants, awareness of risk factors was significantly better in those who were married ( β=0.16, P<0.001), divorced ( β=0.05, P=0.039), or widowed ( β=0.06, P=0.027); compared to participants with monthly income<3 000 yuan, the subjects with monthly income from 5 000 to 10 000 yuan ( β=0.07, P=0.020) had a higher knowledge score; compared to participants with no family history of gastric cancer, subjects with family history had higher knowledge level of risk factors for gastric cancer ( β=0.06, P=0.029). Conclusion:The knowledge levels of risk factors for gastric cancer are generally low among residents in Shijiazhuang city. Educational initiatives are required to improve the awareness of risk factors for gastric cancer, and interventions need to be implemented concurrently to change unhealthy behaviors among residents in Shijiazhuang city.

Objective To explore the effects of lung rehabilitation practice on the quality of life and pulmonary function of chemotherapy patients with lung cancer in low lung function. Methods From October 2016 to October 2017, 90 patients with lung cancer in low lung function who were undertaking chemotherapy regularly in the First Affiliated Hospital of Harbin Medical University were randomly divided into control group and observation group, with 45 cases in each group. The patients of both the groups were given chemotherapy nursing and health education, while the observation group received pulmonary rehabilitation intervention on the basis of routine nursing. The quality of life and pulmonary function of the patients in the two groups were compared before and 4 weeks after the intervention. Results Before the intervention, there was no significant difference in the quality of life and pulmonary function between the two groups (P＞ 0.05). After 4 weeks' intervention, the total score and the score of each dimension of the life quality scale of the observation group were all better than those of the control group, while the BODE index (Body-mass index, airflow Obstruction, Dyspnea, and Exercise) was lower than that of the control group. All the differences were statistically significant (P＜ 0.05). Conclusions Pulmonary rehabilitation is an effective way to improve the quality of life and lung function of lung cancer patients with low lung function during chemotherapy, which is worthy of clinical promotion.