Objective To modify HLA-A2.1 restricted epitope of tyrosinase related protein-2 (TRP-2) (180-188) of human melanoma differentiation antigen and enhance HLA-A2.1 molecule its affinity to. Methods The TRP-2 (180-188) nave epitope was altered with the predominant amino acid for the primary anchor residues and auxiliary anchor residues. Altered peptide ligands (APLs) were scanned by polynomial method, the quantitative motif method and OSAR methods. The analogues their affinity to HLA-A2.1 molecule were assayed. Results Compared with the nave peptide, APLs had stronger affinity to HLA-A2 molecules. Conclusion Our results suggest that altered nave epitope with P1 (Y) and/or P2(L, M) can enhance affinity to HLA-A2 molecule. However, the immunogical efficacy of APLs needs to be identified in studies in vitro and in vivo.

Objective:To determine bacteriostatic abilities ofArtemisia argyi extracts,and to explore the effect ofArtemisia argyi extracts on oral ulcer in rats.Methods:We extracted the mixture ofArtemisia argyi volatile oils and water-extraction by leaching method and evaluated the anti-microbial effect ofArtemisia argyi extracts on common oral floras in vitro.The rat cheeks were burnt by NaOH to establish the models of oral ulcer.The curative effects of crude drug of Artemisia argyi extracts at 2.0,1.0,0.5 g/mL on oral ulcer in rats were evaluated by measuring the oral ulcer healing time.Serum TNF-α level and expression of proliferating cell nuclear antigen (PCNA) were analyzed by ELASA and immunohistochemical staining.Results:Artemisia argyi extracts obviously inhibited the Staphylococcus aureus and Streptococcus.NaOH-made oral ulcer in rats were successfully established.The crude drug at 2.0 and 1.0 g/mL obviously reduced healing time,significantly inhibited the release of TNF-α,and improved the PCNA level in the ulcer tissues (All P＜0.01).The extracts obviously reduced the local inflammatory reaction and promoted tissue repair of oral ulcer.Conclusion:Artemisia argyi extracts promote tissue repair of oral ulcer via inhibiting bacterial growth,reducing the release of TNF-α and improving the PCNA level.

Objective To investigate the value of serum IgG4 level in the differential diagnosis of IgG4-related pancreatic and hepatobiliary disease (IgG4-PHD) and non-IgG4-related disease (non-IgG4-RD). Methods Clinical data were collected from 491 patients who were hospitalized and 50 individuals who underwent physical examination in Huaian No. 1 People's Hospital Affiliated to Nanjing Medical University, Subei People's Hospital, and The First Affiliated Hospital of Xuzhou Medical University from August 2014 to April 2021. The 491 patients were divided into IgG4-PHD group with 20 patients, non-IgG4-RD autoimmune disease group with 431 patients (104 patients with systemic lupus erythematosus, 79 with rheumatoid arthritis, 174 with Sjogren's syndrome, 16 with ankylosing spondylitis, 11 with scleroderma, 4 with adult-onset Still's disease, 30 with myositis, 3 with psoriasis, and 10 with primary sclerosing cholangitis), and malignant pancreatic/hepatobiliary tumor group with 40 patients, and the 50 individuals undergoing physical examination were enrolled as healthy control group. Scattering immunoturbidimetric assay was used to measure serum IgG4 concentration. The two-sample Mann-Whitney U test was used for comparison of normally distributed continuous data between groups, and the Fisher's exact test was used for comparison of categorical data between groups. The receiver operating characteristic (ROC) curve was plotted to determine the optimal cut-off value of serum IgG4 in the diagnosis of IgG4-PHD. Results The IgG4-PHD group had a significantly higher serum IgG4 level than the non-IgG4-RD autoimmune disease groups, the malignant pancreatic/hepatobiliary tumor group, and the healthy control group (all P < 0.05), and the Sjogren's syndrome group had a significantly lower serum IgG4 level than the healthy control group ( Z =2.958, P < 0.05). With serum IgG4 ≥1.35 g/L and IgG4 ≥2.01 g/L as the cut-off values, the IgG4-PHD group had a significantly higher positive rate than the non-IgG4-RD autoimmune disease group and the healthy control group (all P < 0.05). The ROC curve analysis showed that IgG4 had an area under the ROC curve of 0.980 in the differential diagnosis of IgG4-PHD and non-IgG4-RD autoimmune diseases, with a sensitivity of 100.00% and a specificity of 94.00% at the optimal cut-off value of 2.21 g/L. Conclusion Serum IgG4 level may also increase in non-IgG4-RD autoimmune diseases, while the cut-off value of 2.21 g/L can improve the differential diagnosis of IgG4-PHD and non-IgG4-RD autoimmune diseases, which requires further verification in clinical practice.

Objective: To explore the role of TLR 9 in intrauterine transmission of hepatitis B virus (HBV) through blood pathway and placenta. Methods: Epidemiological investigation was carried out in 290 HBsAg positive parturients and 45 normal parturients (control group) in Northwest Women and Children Hospital of Shaanxi Province. Enzyme-linked immunosorbent assay (ELISA) was used to detect five serological makers of hepatitis B and TLR 9 levels in peripheral blood of pregnant women and newborns. HBV DNA was detected by real-time fluorescence quantitative PCR. Detection of TLR 9 expression in placenta by immunohistochemical method. A case-control study was conducted to analyze the difference of TLR 9 levels in placenta and peripheral blood of HBsAg- positive pregnant women with intrauterine transmission of HBV. Results: The incidence of dominant HBV infection (DBI), occult HBV infection (OBI) and intrauterine transmission of HBV were 9.28% (27/291), 40.21% (117/291) and 49.48% (144/291) respectively. (1) The level of TLR 9 in peripheral blood of HBsAg-positive parturients, non-HBV intrauterine transmission (NBIT) group and OBI group were significantly lower than that of control group (P<0.001). The level of TLR 9 in DBI group was significantly higher than those in NBIT group and OBI group (P=0.000). (2) The TLR 9 level in HBeAg-negative group was significantly lower than that in HBeAg-positive parturients in OBI group (P=0.01). (3) With the increased severity of intrauterine transmission of HBV in each HBV DNA load group, the TLR 9 level in maternal peripheral blood increased significantly (P<0.05). (4) With the increased severity of intrauterine transmission of HBV, the levels of TLR 9 increased significantly in antiviral therapy, immunoglobulin injection and non-hepatitis B vaccine groups (P<0.05). (5) The expression of TLR 9 in placenta tissues with DBI group was significantly higher than that in OBI group and NBIT group (P<0.05). Conclusions HBV can inhibit the secretion of TLR 9 in parturient to some extent, but HBeAg can stimulate the secretion of TLR 9. However, with the increased severity of intrauterine transmission of HBV, the level of TLR 9 in parturients is increased by intra-group cross-differentiation. Therefore, TLR 9 is not an independent marker for screening and grouping, but it can be used as an reference indicator for the monitoring and management of HBsAg-positive parturients.

Objective: To investigate the expression of IL-18 in peripheral blood of HBsAg positive parturients in intrauterine transmission of HBV. Methods: A case-control study was conducted in 282 HBsAg positive parturients and 43 health parturients (control group) in Northwest Women and Children Hospital of Shaanxi Province. Enzyme-linked immunosorbent assay (ELISA) was used to detect five serological makers of hepatitis B, real time PCR was used to detect HBV DNA, and flow liquid chip method was used to detect IL-18 levels in peripheral blood of parturients and newborns. Results: The incidence of dominant HBV infection (DBI), occult HBV infection (OBI) and intrauterine transmission of HBV were 8.42% (24/285), 40.00% (114/285) and 48.42% (138/285), respectively. The level of IL-18 in peripheral blood of HBsAg-negative parturients were significantly lower than those of HBsAg-positive parturients (P=0.001), non-HBV intrauterine transmission (NBIT) group (P=0.001) and OBI group (P<0.001). The level of IL-18 in HBeAg negative group was significantly lower than that in HBeAg positive group (P=0.023). When HBV DNA load was ≥10³ copies/ml, the level of IL-18 was significantly higher than that in HBsAg-negative group (P<0.01). With the increase of HBV DNA load in maternal blood, the level of IL-18 increased (P=0.024). When HBV DNA load was 10³-10⁶ copies/ml, the level of IL-18 in DBI group was significantly lower than that in NBIT group (P=0.022), and increased with the increase of HBV DNA load in maternal blood (P=0.016). With the increased severity of intrauterine transmission of HBV, the level of IL-18 in non-hepatitis B vaccine group decreased significantly (P=0.044). The level of IL-18 in non-hepatitis B vaccine group and immunoglobulin injection group was significantly higher than that in NBIT group (P<0.05). Multivariate analysis showed that the linear relationship between maternal HBeAg status and maternal IL-18 levels had statistical significance (P=0.01). Conclusions IL-18 is a higher level balance regulator of Th1/Th2 immune network. Monitoring the level of IL-18 in HBsAg-positive parturients can be used not only for predicting the probability of DBI and OBI, but also as an intervention mean, especially for those who are HBeAg-positive and had HBV DNA load ≥10³ copies/ml, to improve maternal cellular immune function, which is conducive to interrupting intrauterine transmission and providing a theoretical basis for the prevention and control of HBV intrauterine transmission.

Objective: To investigate the influence of IFN-γ and IL-12 levels in prenatal peripheral blood of HBsAg-positive parturients on intrauterine transmission of hepatitis B virus (HBV). Methods: A case-control study was conducted in 282 HBsAg positive parturients and 43 health parturients (control group) in Northwest Women and Children Hospital of Shaanxi Province. Enzyme-linked immunosorbent assay (ELISA) was used to detect five serological makers of hepatitis B in peripheral blood of parturients. HBV DNA was detected by real-time fluorescence quantitative PCR. Detection of cytokines IFN-γ and IL-12 levels were conducted with liquid chip-based flow cytometry method. The serum levels of five serological markers of hepatitis B and HBV DNA in 285 newborns were detected within 24 hours after birth. Results: The incidence of intrauterine dominant infection (DBI), occult infection (OBI) and intrauterine transmission of HBV in HBsAg positive parturients were 7.37% (21/285), 40.70% (116/285) and 48.07% (137/285), respectively. The level of IFN-γ in peripheral blood of HBsAg-negative parturients was significantly lower than those of HBsAg-positive parturients (t=-2.55, P=0.011), NBIT group (t=-2.54, P=0.012) and OBI group (t=-2.33, P=0.021). In HBV DNA load of 10³-10⁶ copies/ml group, the levels of IFN-γ in the DBI group were significantly lower than those in OBI group and NBIT group (P<0.01). The level of IFN-γ in maternal peripheral blood decreased significantly with the increased severity of intrauterine transmission of HBV (χ²=6.40, P=0.041). In the antiviral treatment group, the level of IL-12 in maternal peripheral blood decreased significantly with the increased severity of intrauterine transmission of HBV (χ²=8.90, P=0.012). Multivariate analysis showed that there was a significant linear relationship between maternal IFN-γ level and maternal age, placenta previa and hepatitis B vaccine injection (P<0.05). The linear relationship between the level of maternal IL-12 and the mode of rupture and hepatitis B vaccine injection had statistical significance (P<0.05). Conclusions HBV can stimulate the expression of IFN-γ and inhibit the secretion of IL-12 in pregnant and lying-in women, but the expression of IFN-γ in HBsAg-positive parturients showed intra-group differentiation, and the maternal level of IFN-γ will decrease in HBeAg-positive and high-HBV DNA loadstatus. Increasing the levels of IFN-γ and IL-12 in HBsAg-positive parturients is beneficial to block intrauterine transmission of HBV, especially DBI.