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 establish a mouse model of infection with the minimum lethal dose of Vibrio vulnificus(V.vulnificus)and to evaluate the protective efficacy of inactivated whole-cell(IWC)vaccine using this model.Methods A mouse model of lethal-dose infection was established by intraperitoneal injection of different doses of V.vulnificus.Bacterial colonization in the organs was detected with tissue homogenate plating,and pathological changes in the organs were observed after tissue section staining.Flow cytometry was used to detect immune cell responses after liver tissues were digested into single-cell suspension.IWC vaccine of V.vulnificus was prepared,and the mice were immunized through different routes to observe the protective efficacy of the vaccine.Results A mouse model of infection with the minimum lethal dose at 1×106 CFU of V.vulnificus was successfully established.After infection,the bacteria were mainly colonized in the liver of mice and caused severe pathological damages.Compared with the uninfected mice,the proportion of neutrophils in the liver was significantly increased in the infected mice,whereas the proportions of B cells and T cells were correspondingly decreased(P<0.05).A single intramuscular or intraperitoneal injection of the IWC vaccine could protect the mice effectively against lethal infection of V.vulnificus in 7 d later(P<0.01),although the level of serum IgG having no significant increase.Conclusion A mouse model of lethal-dose infection with V.vulnificus is successfully established,with histopathological characteristics.The IWC vaccine of V.vulnificus rapidly mediates immune protection in this model probably independent of IgG.

The occurrence of benign prostate hyperplasia(BPH)was related to disrupted sex steroid hormones,and metformin(Met)had a clinical response to sex steroid hormone-related gynaecological disease.How-ever,whether Met exerts an antiproliferative effect on BPH via sex steroid hormones remains unclear.Here,our clinical study showed that along with prostatic epithelial cell(PEC)proliferation,sex steroid hormones were dysregulated in the serum and prostate of BPH patients.As the major contributor to dysregulated sex steroid hormones,elevated dihydrotestosterone(DHT)had a significant positive rela-tionship with the clinical characteristics of BPH patients.Activation of adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK)by Met restored dysregulated sex steroid hormone homeostasis and exerted antiproliferative effects against DHT-induced proliferation by inhibiting the formation of androgen receptor(AR)-mediated Yes-associated protein(YAP1)-TEA domain transcription factor(TEAD4)heterodimers.Met's anti-proliferative effects were blocked by AMPK inhibitor or YAP1 over-expression in DHT-cultured BPH-1 cells.Our findings indicated that Met would be a promising clinical therapeutic approach for BPH by inhibiting dysregulated steroid hormone-induced PEC proliferation.

Objective: To investigate the current status of HIV-related stigma among college students in Guilin,and to explore the impact of perspective-taking and empathy on HIV-related stigma among college students. Methods: The stratified cluster sampling method was used to select college students from 4 universities in Guilin city. Stratified randomization was used to assign all subjects into a control group (410) and an experimental group (396) for randomized controlled trials,and the HIV-related Stigma Questionnaire and the Basic Empathy Scale were used before and after the intervention. Results: Before the intervention of perspective-taking,there were no statistically significant differences between two groups in subscale scores and total scores of HIV-related stigma (t=0.80，0.35，-0.62，-0.10，P>0.05); However, signiticant differences in subscale scores and total score in HIV-related stigma were found after intervention (t=3.53，2.21，2.30，3.98，P<0.05). There was no statistically singnificant in the scores of all dimensions of empathy level and the total score before the intervention (t=0.10,-0.27,-0.08,P>0.05), dimensional score and total score in empathy were statistically significant after intervention (t=-2.15，-3.06，P<0.05). Empathy played an intermediary role of 14.08% between opinion selection and HIV stigma. Conclusion HIV-related stigma exists among college students in Guilin,and perspective-taking intervention effectively reduces its AIDS stigma. Empathy plays an intermediary role between them.

As the proceeding of the Belt and Road, TCM industry in Russia develops steadily. Acupuncture and Tuina therapies have been accepted by the local people, while Chinese medicine is still around the acceptance. Right now, Russian economy is on a recovering stage, so it's good time for TCM to carry service trading during this time. TCM development in Russia need to focus on the input of medical resources, managements and regulations, proceeding TCM teaching work and bringing along TCM service trade by acupuncture therapy.

OBJECTIVE: To observe the effects of maternal low-level decabromodiphenyl( BDE209) exposure on nervous system and secretion of thyroid hormones in offspring mice. METHODS: Sixty-four specific pathogen free female,aged 4 weeks Kunming mice were used. These mice were randomly divided into control group and low,medium and high exposure groups after successful mating was confirmed. The rats of control group were fed with 0. 01 L/kg body mass of peanut oil.The maternal mice in the experimental groups were given BDE209 at doses of 50,100 and 300 μg/kg body mass by oral gavage once per day. Continuous exposure was given until 21 days after birth of offspring,the exposure model from gestation to lactation was established. At the end of the exposure,10 mice of each group including half female and half male were randomly selected and the body mass and growth development status were observed. Morris water maze was used to examine the learning and memory ability in offspring mice. The serum levels of total triiodothyronine( TT3),free triiodothyronine( FT3), total tetraiodothyronine( TT4) and free tetraiodothyronine( FT4) were measured by chemiluminescence immunoassay. The levels of glutathione transferase( GST), superoxide dismutase( SOD) and malondialdehyde( MDA) level in hippocampus were measured by colorimetry. RESULTS: The escape latency of the medium exposure group was longer than that of the control group( P < 0. 05). The escape latency of high exposure group was longer than that of control group,low exposure group and medium exposure group( P < 0. 05). The time of quadrant movement and number of crossing the platform in offspring rats in high exposure group were less than that of the control group and the low and medium exposure groups( P < 0. 05). The serum levels of TT3,FT3,TT4 and FT4 increased,the activities of GST and SOD in hippocampus tissue decreased,the MDA level increased with the increasing exposure dose( P < 0. 05).CONCLUSION: Maternal low-level BDE209 exposure can result in decrease the learning and memory ability of offspring mice. It also can increase the serum thyroid hormone level and induce oxidative stress injury in hippocampus in a dose dependent manner in offspring mice.

Objective To investigate the effect of low dose of BDE209 on thyroid hormone and thyroid hormone metabolism enzyme-iodothyroninedeiodinases Ⅱ(D2) in off spring mice after pregnancy exposure .Methods Total 64 adult SPF female Kun-ming mice were randomly divided into 4 groups ,which treated with oral gavage of 0 ,50 ,100 ,300 μg · kg -1 · d-1 dose of BDE209 after successful pregnancy ,the exposure continue to 21 days after delivery .10 mice was randomly selected in each offspring group and get the peripheral blood and brain sample ,the serum thyroid hormones level ,oxidative damage and the expression of D2 mRNA in brain were detected .Results Compared with the control group(0μg · kg -1 · d-1 dose of BDE209) ,the TT4 ,TT3 ,FT4 and FT3 levels of offspring mice increased significantly in every exposure group (P< 0 .05);antioxidant enzyme glutathione-S transferees (GST) ,superoxide dismutase (SOD) activity decreased with the BDE209 dose increase (P<0 .05) ,and malondialdehyde (MDA) level increased (P<0 .05);the D2 mRNA relative expression of brain in middle(100μg · kg -1 · d-1 dose of BDE209) and high(300μg · kg -1 · d-1 dose of BDE209) dose group decreased when compared with control group (P< 0 .05) .Conclusion Low level of BDE209 exposure in pregnancy resulted in the increasing of thyroid hormone levels in offspring mice ,which may cause oxidative damage and decrease expression of D2 mRNA in the brain .