Objective: To investigate the enhancement effect of Xinjiang wild Artemisia rupestris L. crude polysaccharides (WARCP) as an adjuvant on ovalbumin (OVA) vaccine in mice immunized intramuscularly. Methods: ICR mice were randomly divided into 6 groups (5 per group), including 9 g/L NaCl group (blank control), OVA group (10 μg OVA), low dose WARCP/OVA group (OVA+50 μg WARCP), medium dose WARCP/OVA group (OVA+200 μg WARCP), high dose WARCP/OVA group (OVA+400 μg WARCP), and aluminum adjuvant (Alum)/OVA group (positive control group, OVA+100 μg Alum). ICR mice were immunized intramuscularly and weighted. The OVA-specific antibodies and subtypes in serum were detected by enzyme linked immunosorbent assay (ELISA). T cells subsets from spleen and lymph nodes were detected by flow cytometry. Results: The medium-dose WARCP/OVA group enhanced IgG and IgG1 levels and increased early antibody levels (all P<0.05). The medium-dose WARCP/OVA group and the high-dose WARCP/OVA group significantly enhanced IgG2a levels (all P<0.05), but the difference was not statistically significant comparing with Alum/OVA group (P>0.05). The low-dose WARCP/OVA group enhanced the percentage of CD4⁺ T cells in spleen and CD4⁺ T, CD8⁺ T, CD4⁺CD44⁺ T cells in lymph nodes (all P<0.05). The medium dose WARCP/OVA group and the high dose WARCP/OVA group enhanced the CD4⁺ T, CD8⁺ T, CD4⁺CD44⁺ T, CD8⁺CD44⁺ T cells in spleen and CD8⁺CD44⁺ T cell in lymph nodes (all P<0.05). Conclusions Plant-derived WARCP as an OVA protein vaccine adjuvant can enhance cellular immunity and humoral immunity, and it is safe and reliable. The results in this study provide a theoretical basis for the popularization and application of WARCP.

Objective:To investigated the effects of storage temperature and storage state on the immunoregulatory activities of wild Artemisia rupestris L. crude polysaccharides (WARCP) and cultivated Artemisia rupestris L. crude polysaccharides (CARCP). Explore the optimal storage conditions for WARCP and CARCP.Methods:WARCP and CARCP were stored at different temperatures (4, -20 and -80 ℃) and in different states (powder and solution) for 6 months. Different doses (10, 50, 100 μg/ml) of WARCP and CARCP were used to stimulate mouse bone marrow dendritic cells (DCs) for 24 h in vitro. The lipopolysaccharide with a dose of 100 ng/ml were used as positive control, and RPMI-1640 medium was used as negative control. Flow cytometry was used to detect the proportions of CD40 and CD86 on the surface of DCs.Results:At different temperatures, the differences in immunomodulatory activity of WARCP and CARCP stored in powder state and WARCP stored in solution state were not statistically significant (all P>0.05). The immunomodulatory activities of CARCP stored in solution state at -20 ℃ and -80 ℃ were significantly better than that stored at 4 ℃ (all P<0.05). Conclusions:After 6 months of storage, the immunomodulatory activity of WARCP and CARCP is not affected by the storage state, and can maintain good immunomodulatory activity at different storage temperatures.

Objective To compare the immunopotentiating effects of polysaccharides extracted from wild/cultivate Cistanehe deserticola (WCDPS/CCDPS) in Xinjiang. Methods ICR mice were subcu-taneously injected twice with different doses(low,medium and high) of WCDPS and CCDPS in combination with ovalbumin (OVA). OVA-specific antibody IgG,as well as IgG1 and IgG2a subtypes, was determined by ELISA. OVA-specific lymphocyte proliferation was measured by MTT. Expression of CD4+T and CD8+T cells was analyzed by flow cytometry. Results Both WCDPS and CCDPS could significantly improve the production of OVA-specific IgG,IgG1 and IgG2a,promote the proliferation of OVA-specific lymphocytes and increase the expression of CD4+T and CD8+T cells(all P<0.05) with no significant difference between them at the same dosages (P>0.05). WCDPS and CCDPS had no influence on the body weight of mice after im-munization. Conclusion WCDSP and CCDPS could significantly enhance the OVA-specific humoral and cellular immune responses with no statistical difference and are characterized by high safety.

Objective To study the effects of aqueous extracts of cultivated Cistanche deserticola Y. C. Ma (AECCD) on T cell responses and the duration of antibody response and to investigate its immunoen-hancing activities in mice. Methods Two batches of female ICR mice were used in this study with 30 from each batch. Each batch of mice was randomly divided into six groups (n=5). Low, medium and high doses of AECCD in combination with ovalbumin ( OVA) were used to set up three experimental groups, while 0. 9% NaCl, OVA alone and aluminium adjuvant were respectively used as blank, negative and positive controls. All mice were intramuscularly injected twice at an interval of two weeks. Flow cytometry was used to detect the ex-pression of T lymphocyte subsets, cytokines and surface molecules of dendritic cells (DC). Indirect ELISA was used to detect IgG antibody levels. Results AECCD could significantly increase the percentage of CD4+and CD8+T lymphocytes in spleen (P<0. 05), up-regulate the expression of CD4+CD44+and CD8+CD44+effector T lymphocytes (P<0. 05), promote the secretion of IFN-γ in T lymphocytes and enhance the expression of CD40 and CD80 on the surface of DC (P<0. 05). ELISA results showed that high-dose AECCD could significantly prolong the duration of IgG antibody response induced by OVA (P<0. 05). Conclusion AECCD could en-hance the T lymphocyte immune response induced by OVA and keep it maintained at a high level, which might help to improve the body′s immune response.

Objective To investigate the immunopotentiating effects of cultivated Cistanche deser-ticola (C.deserticola) crude polysaccharides (CCDCP) as an adjuvant on the model antigen ovalbumin (OVA). Methods Low,medium and high doses of CCDCP in combination with OVA were intramuscularly injected twice into ICR mice at an interval of two weeks,respectively. Aluminum adjuvant was used to set up positive control group. Levels of IgG,IgG1 and IgG2a antibodies were detected by ELISA. Splenocyte prolif-eration was detected by MTT assay. Growth conditions of the immunized mice were observed. Results IgG level was significantly increased in the high dose group 7 days after the first immunization(P<0.05),espe-cially on the 21st and 28th days (P<0.01) as compared with that of the aluminum adjuvant group. High dose of CCDCP in combination with OVA significantly up-regulated the levels of IgG1 and IgG2a in mice as compared with immunization with OVA alone (P<0.05). Moreover, IgG2a level in mice immunized with high dose of CCDCP and OVA was higher than that of the aluminum adjuvant group(P<0.05). Splenocyte proliferation was significantly enhanced in the medium and low dose groups in comparison with that of the OVA group (P<0.05) and the aluminum adjuvant group (P<0.01). No significant difference in mouse body weight was observed in different groups(P>0.05). Conclusion CCDCP as an adjuvant of OVA pro-tein vaccine can enhance Th1 and Th2 immune responses,especially the early antibody production and Th1 immune response. These results will provide some information for further studies of CCDCP as a vaccine ad-juvant.

Objective:To investigate the effect of wild Cistanche deserticola crude polysaccharides (WCDCP) on the immune response of ovalbumin (OVA).Methods:42 ICR mice were randomly divided into the 9 g/L NaCl group (blank sample), WCDCP group (400 μg WCDCP), OVA group (10 μg OVA), low-dose WCDCP/OVA group (100 μg WCDCP+10 μg OVA), medium-dose WCDCP/OVA group (400 μg WCDCP+10 μg OVA), high-dose WCDCP/OVA group (800 μg WCDCP+10 μg OVA), and aluminum adjuvant/OVA group (positive concentration, 200 μg aluminum adjuvant+10 μg OVA). Each group included 6 mice. The mice were immunized by using two-point injection into the muscles of the hind legs of the mice. A total of 2 immunizations, and the immunization was boosted once every 2 weeks after the initial immunization. The body weight of the mice was weighed 7, 14, 21, and 28 days after the initial immunization of the mice, and the changes in body weight and growth status of the mice were observed. The IgG antibodies and antibody fractions were detected by indirect enzyme-linked immunosorbent assay. Twenty-one days after the initial immunization, the spleen lymphocyte proliferation level was detected by the thiazole blue method. Flow cytometry was used to detect the proportion of T cell subsets in the spleen and lymph nodes.Results:At 7 days after the initial immunization, the serum IgG antibody level (0.597 6±0.110 7) in the high-dose WCDCP/OVA group was significantly higher than (0.254 4±0.074 8) of the OVA group ( P<0.05). At 28 days after the initial immunization, the serum IgG, IgG1 and IgG2a antibody levels in the high-dose WCDCP/OVA group were higher than those in the OVA group, the comparison respectively were 0.972 3±0.243 8 vs. 0.389 2±0.077 4 ( P<0.05), 1.156 0±0.088 4 vs. 0.612 6±0.059 7 ( P<0.001), 1.648 0±0.103 9 vs. 0.557 2±0.181 5 ( P<0.001), and the differences were statistically significant. High-dose WCDCP can significantly promote the proliferation of spleen cells induced by concanavalin A ( P<0.001) and lipopolysaccharide ( P<0.05). High-dose WCDCP/OVA group can significantly stimulate the activation ratio of T cell subsets in the spleen. The proportion of CD3 +CD8 + T cells in the high-dose WCDCP/OVA group [(10.83±0.44)%] was significantly higher than that in the OVA group[(6.76±0.58)%] ( P<0.01), and the proportion of CD3 +CD4 + T cells [(28.17±1.67)%] was also significantly higher than that in the OVA group [(19.17±2.73)%] ( P<0.05). WCDCP had no effect on body weight (all P>0.05) and growth of mice. Conclusions:WCDCP can enhance humoral immune response and cellular immune response, and has no side effect on the growth of mice, suggesting that WCDCP can be used as a potential adjuvant for OVA.

Objective:To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT).Methods:A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ 2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results:A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment.Conclusion:ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.

Objective To investigatethe relationship between gastroesophageal reflux disease (GERD) symptoms and clinicopathological characteristics, p53 expression, and survival of Chinese patients with esophageal adenocarcinoma. Methods A total of 3882 patients with esophageal adenocarcinoma, 970 matched patients with esophageal squamous cell carcinoma, and 970 matched patients with gastric cardia adenocarcinoma were included to compare the incidence of GERD symptoms. Patients with esophageal adenocarcinoma were divided into two groups according to the presence and absence of GERD symptoms. The differences in clinicopathological characteristics and p53 expression between the two groups were compared by chi-square test, and the differences in survival between the two groups were compared by landmark analysis. Results The incidence of GERD symptoms increased successively in esophageal squamous cell carcinoma, esophageal adenocarcinoma, and gastric cardia adenocarcinoma. In patients with esophageal adenocarcinoma, the proportions of male, less than 65 years old, lower thoracic tumors, tumors without squamous cell carcinoma, poorly differentiation, early tumors (stage 0-I), and p53-positive tumors in the group with GERD symptoms were higher than those in the group without GERD symptoms; moreover, the long-term survival (≥15 years) was better. Conclusion GERD symptom is an important clinical sign of esophageal adenocarcinoma. It is closely related to specific clinicopathological characteristics, p53 overexpression, and good long-term prognosis.