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Background & objectives: Oral squamous cell carcinoma (OSCC) is widely prevalent in the Indian subcontinent mainly due to habit-associated aetiologies. Immune regulation and angiogenesis are the part of tumourigenesis that play a crucial role in metastasis and survival. However, the concurrent expression of vascular endothelial growth factor (VEGF) and CD3 (immune regulator receptor on T-lymphocyte) in the same OSCC tissue samples has not been reported in the Indian population. The present study evaluated the expression of CD3+ T-cells and VEGF in OSCC tissue samples and studied the clinicopathological correlation and survival analysis in an Indian population. Methods: This was a retrospective study conducted on 30 formalin-fixed and paraffin embedded sections which were histologically diagnosed as OSCC cases comprising of 15 metastatic OSCC and 15 non- metastatic OSCC with available clinical data and survival status. Results: Reduced expression of CD3+ T-cells and increased VEGF expression were observed in metastatic OSCC samples. The correlation of expression of CD3+ T-cells and VEGF with clinicopathological parameters showed a significant association between these markers with age, nodal status, site of the lesion and survival. Interpretation & conclusions: Reduced expression of CD3+ T-cells in OSCC was found to be associated with a significantly poor survival. VEGF was found to be over expressed in metastatic OSCC as compared to that in non-metastatic OSCC. The study findings suggest that the evaluation of CD3 and VEGF in incisional OSCC biopsies can be considered for predicting the survival outcome and metastasis
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@#Periodontitis is a chronic infectious disease that occurs in periodontal support tissues. Plaque microorganisms are its initiating factor, while local inflammation and alveolar bone loss resulting from periodontitis are the most common causes of tooth loss. Interleukin-17 (IL-17), which plays an important role in the immune response to periodontitis, mostly originates from T helper cell 17 (Th17) and γδT cells. In periodontitis, the role of Th17 cells has been demonstrated broadly, but the role of γδT cells was not revealed until recent years. As a highly heterogeneous group of T lymphocytes, γδT cells are considered a link between innate immunity and adaptive immunity. Studies have found that γδT cells are mostly distributed in the oral epithelium near the biofilm, where they can interact with microorganisms to produce IL-17, recruit neutrophils, macrophages, etc., and participate in the host immune response to periodontitis. They also play a role in the association between periodontitis and relevant systemic diseases. In addition, γδT cells have been proven to produce tissue repair-related factors with a protective effect against periodontitis. The possible mechanism of γδT cells in periodontitis is discussed in this review.
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Bilirubin has good anti-inflammatory， antioxidant and immunomodulatory effects， but its poor water solubility and low bioavailability greatly limit its clinical application. Researchers have developed bilirubin into various nanoparticles， which effectively eliminate the limitation of low solubility of bilirubin with the advantage of dosage form， so that they can maximize its pharmacological activities such as anti-inflammatory， anti-oxidation and immune regulation. Bilirubin nanoparticles have great application potential in a variety of gastrointestinal diseases， liver and kidney diseases， skin diseases， autoimmune diseases， islet transplantation and targeted therapy of tumors （both as a direct anti-tumor drug and as a drug delivery system）. The study of bilirubin nanoparticles will promote the clinical application of bilirubin and the development of related new drugs.
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Polysaccharides have significant immunomodulatory activity and have good development value in food and medicine fields. At present, there are many studies on the chemical structure and immune activity of polysaccharides, but the relationship between them of polysaccharides has not been fully explained, which limits the further development and utilization of polysaccharide resources. The immune activity of polysaccharides is closely related to their own structure. This paper systematically summarized the relationship between the relative molecular weight, monosaccharide composition, glycosidic bond types, chemical modification, and advanced conformation of polysaccharides and the immune regulation, aiming to provide references for the profound study of polysaccharide structure-activity relationship and utilization of polysaccharides.
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Monosaccharides/chemistry , Structure-Activity Relationship , Molecular Weight , Antioxidants/pharmacology , Polysaccharides/chemistryABSTRACT
Establish a production line with controllable process and high intelligence, contribute to improve the quality and production efficiency of aconite processed by microwave, and promote the transformation and application of aconite processed by microwave. According to the principle of aconite detoxification and the characteristics of industrial microwave equipment, an industrial production line of aconite processed by microwave was established with diester alkaloids and monoester alkaloids as indicators, and pilot production was carried out. At the same time, the content of active constituents and efficacy were compared with that of the main processed products, such as Shengfupian, Baifupian and Heishunpian. The results showed that the industrial production of aconite processed by microwave can be divided into two stages: "Liquid seal to detoxification - drying and puffing". The content of monoester alkaloids in 10 batches of aconite processed by microwave was 0.071%-0.166% and the content of diester alkaloids was 0.004%-0.016%, which met the relevant requirements of the Chinese Pharmacopoeia in 2020. Compared with Heishunpian and Baifupian, the retention rate of the effective components of aconite processed by microwave was higher. Pharmacological experiments showed that aconite processed by microwave not only retained the anti-inflammatory and analgesic activities of Heishunpian and Baifupian, but also significantly increased the levels of leukocytes and lymphocytes in mice with liver cancer chemotherapy, enhanced the CD4/CD8 ratio in spleen cells of mice (P < 0.05), thus regulating the body's immunity. However, this effect of Baifupian was weak, while Heishunpian and Shengfupian had no such effect. Through the above research, this study established microwave processing line with controllable process and high intelligence, as well produced the aconite processed by microwave with low toxicity and stable quality. It laid a foundation for the industrialized continuous production and clinical positioning of aconite by microwave processed, and provided scientific support for the development and application of microwave technology in the field of traditional Chinese medicine. All animal experiments in this study were reviewed and approved by the Experimental Animal Ethics Committee of Chengdu University of Traditional Chinese Medicine before being carried out (Approval No. 2020-28).
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Functional peptides refer to peptides that are beneficial to life activities or have special physiological activities, also known as bioactive peptides. Oyster is rich in protein and is a good material for developing bioactive peptides, which has great potential as a functional food and great application value in pharmaceutical and medical industry. With the development of modern biotechnology and medical technology, the method innovation of oyster peptide preparation，the absorptivity and biological activity of oyster peptide have been enhanced significantly, which lead to deep recognition of the biological function of oyster peptide and offer the boarder application prospect. The researches on the diversification activities of oyster peptides were summarized in this review, which provided clues and ideas for the development of the oyster peptide applications.
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Helminth infections may trigger host innate and adaptive immune responses. Group 2 innate lymphoid cells (ILC2) are an important factor involved in type 2 immune responses, and produce a large number of T helper 2 cell (Th2) cytokines following stimulation by interleukin (IL)-25, IL-33 and thymic stromal lymphopoietin (TSLP), which play a critical role in parasite clearance and tissue repair. Following helminth infections, autocrine factors, mast cells, enteric nervous system and Th2 cells have been recently found to be involved in regulation of ILC2. Unraveling the role of ILC2 in immune response against helminth infections is of great value for basic research and drug development. This review summarizes the research progress on ILC2 and its role in helminth infections.
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@#To investigate the effects of Shengmai formula (SMF) on tissue damages, serum inflammatory factors and the proportion of innate immunocytes in peripheral blood, sepsis models using either intraperitoneal injection of 20 mg/kg lipopolysaccharide (LPS) or cecal ligation and puncture (CLP) were established.The role of gut microbiota in septic mice during SMF treatment was further investigated.LPS-induced sepsis model was carried out 4 days after daily gavage administration with 0.3 g/kg, 0.6 g/kg, 1.2 g/kg SMF or intraperitoneal injection with 0.6 g/kg SMF.Survival rates of septic mice were determined.Histological evaluations of liver, lung and kidney were analyzed by H&E staining. Serum IL-6, TNF-α, Alanine transaminase (ALT), Aspartate aminotransferase (AST), Blood urea nitrogen (BUN) and Creatinine (Cr) levels were determined.LPS and CLP-induced sepsis models were established, and the proportion of monocytes, macrophages and neutrophils in peripheral blood were analyzed by flow cytometry after gavage administration or intraperitoneal injection of SMF.The therapeutic effects of SMF after antibiotics treatment were further determined, and the therapeutic effects of fecal microbiota from SMF-treated mice were investigated.The results show that LPS-induced sepsis caused death of mice, damages in liver, lung and kidney with increased infiltration of leukocytes and elevated levels of serum IL-6, ALT, AST, BUN and Cr, which were all reversed by gavage administration of SMF.Gavage administration of SMF could significantly reduce the proportion of peripheral macrophages in LPS model and monocytes, macrophages, neutrophils in CLP model.Intraperitoneal injection of SMF showed no therapeutic benefits in septic mice.Depletion of gut microbiota using antibiotics cocktail reversed the therapeutic effects of SMF on sepsis, indicating the involvement of gut microbiota.Fecal microbiota from SMF-treated donors was transplanted into pseudo-sterile recipients, and we found FMT could significantly ameliorate sepsis of recipients.These results showed that gavage administration of SMF reduced serum inflammatory factors and alleviated tissue damages in septic mice by regulating gut microbiota. This study provides a theoretical basis for the treatment of clinical sepsis with SMF.
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Chronic graft-versus-host disease (cGVHD) is the main complication after allogeneic hematopoietic stem cell transplantation, which is also the major cause of non-relapse -related death. Due to its complex pathophysiological process, the response rate of conventional glucocorticoids combined with immunosuppressants is less than 50%. Second-line therapy should be given for patients with glucocorticoid-resistant cGVHD. Nevertheless, no consensus has been reached on current second-line therapy and the therapeutic effect is relatively poor. Mesenchymal stem cell (MSC) is one of the most common adult stem cells. Due to multi-dimensional and multi-target immune regulating function, MSC has been widely applied in the prevention and treatment of cGVHD. In addition, accumulated studies have confirmed the safety and efficacy of MSC in the treatment of cGVHD, which is expected to become a novel strategy for the prevention and management of cGVHD. In this article, research progress, mechanism and existing problems of prevention and treatment of cGVHD by MSC were reviewed, aiming to provide novel ideas for optimizing therapeutic regimens of MSC and enhancing the prevention and treatment effect of cGVHD in subsequent research.
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ObjectiveTo observe the effect of Qinggan Jianpi Huoxue prescription（QGJPHXP） on the polarization of M1/M2 macrophages in rats with hepatic fibrosis induced by carbon tetrachloride（CCl4）. MethodA rat hepatic fibrosis model was established by intraperitoneal injection of 40% CCl4-olive oil suspension twice a week at the dosage of 2.0 mL·kg-1 for 8 weeks. After the model was successfully established， these rats were randomly divided into the model group， QGJPHXP group（32.084 g·kg-1） and Biejiajian pills（BJJP） group（0.925 5 g·kg-1）， with 12 rats in each group. The blank group was injected intraperitoneally with the same amount of olive oil. The rats in the administration groups were given the corresponding solution according to the dose， and the blank and model groups were given the same dose of purified water， once a day. After 4 weeks of continuous administration， the liver tissues of rats were taken and stained with hematoxylin-eosin（HE） and Masson to observe the pathological changes. The serums were collected to detect the alanine aminotransferase（ALT） and aspartate aminotransferase（AST） levels. Interleukin（IL）-6， IL-12， IL-10， IL-1β， transforming growth factor-β1（TGF-β1） and tumor necrosis factor-α（TNF-α） levels in liver tissues were measured by enzyme-linked immunosorbent assay（ELISA）. The expression levels of CD86 and CD206 were detected by immunohistochemistry（IHC）. Western blot and real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） were used to detect the protein and mRNA expression levels of inducible nitric oxide synthase（iNOS）， arginase-1（Arg-1）， phosphorylated p38 mitogen-activated protein kinase（p-p38 MAPK）， nuclear transcription factor-κB p65（NF-κB p65） in liver tissues of rats. ResultCompared with the blank group， the hepatic cell plate was irregularly arranged, and local inflammatory cell infiltration and fibrous hyperplasia were observed， while the serum levels of ALT and AST were significantly increased in the model group（P<0.01）， and IL-1β， IL-6， IL-12， TGF-β1， TNF-α， CD86， CD206， iNOS， p-p38 MAPK，p38 MAPK and NF-κB p65 levels in liver tissues were obviously increased（P<0.05， P<0.01）， while the levels of IL-10 and Arg-1 were obviously decreased（P<0.05， P<0.01）. Compared with the model group， QGJPHXP group reduced the degree of liver cell fibrosis，and serum levels of ALT and AST（P<0.01）， and IL-1β， IL-6， IL-12， TGF-β1， TNF-α， CD86， iNOS， p-p38 MAPK， p38 MAPK， and NF-κB p65 levels in liver tissues were obviously decreased（P<0.05， P<0.01）， the levels of IL-10， CD206 and Arg-1 were obviously increased in the QGJPHXP group（P<0.05， P<0.01）. ConclusionQGJPHXP has ability to inhibit the activation of pro-inflammatory M1 macrophages， induce the secretion of anti-inflammatory cytokines by M2 macrophages， reduce the release of pro-fibrogenic cytokines， and promote the macrophage polarization of M1 to M2 in liver for tissue repair， thereby serving as an anti-inflammatory and anti-hepatic fibrosis drug.
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Intestinal flora plays an important role in maintaining the body's immune balance, digestion, and nutrient absorption. Thyroid function mainly includes the synthesis and secretion of thyroid hormones and the regulation of metabolic balance in the body. When the body’s thyroid hormone synthesis and secretion are too little or too much, it will lead to thyroid diseases. Studies have shown that intestinal flora may affect the occurrence and development of thyroid diseases by participating in thyroid metabolism, immune regulation, cross immunoreaction, and interaction with thyroid-related predisposing factors. Understanding the structural changes and functional mechanisms of intestinal flora during the occurrence and development of thyroid diseases can provide a new basis for the diagnosis and treatment of thyroid diseases and improvement of clinical prognosis.
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Inflammatory bowel diseases (IBD) are a kind of non-specific inflammatory disease that occurs in gastrointestinal tract. Abnormal immune regulation is a key factor in its pathogenesis. The acquired immune regulation is mediated by helper T cells (Th), which is reported to play an important role in the pathogenesis of IBD. Th17 is a subtype of CD4 + T cells that could specifically produce interleukin-17 (IL-17) and other related cytokines. In this paper, we review the immune modulation of Th17 and its related cytokines in IBD.
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ABSTRACT Interferon-gamma (IFN-γ) plays a crucial role in viral infections by preventing viral replication and in the promotion of innate and adaptive immune responses. However, IFN-gamma can exert distinct effects in different persistent viral infections. The long-term overproduction of IFN-γ in retroviral infections, such as the human immunodeficiency virus (HIV), human T-lymphotropic virus type 1 (HTLV-1), and human endogenous retroviruses (HERVs), resulting in inflammation, may cause neuronal damage. This review is provocative about the role of IFN-γ during persistent retroviral infections and its relationship with the causation of some neurological disorders that are important for public health.
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Objective To investigate the roles of inducible costimulatory molecules (ICOS) and related cytokines in the immune regulation of Echinococcus granulosus infections in mice. Methods Eighty BALB/c mice (weight 18–22 g) were divided into the control and infection groups, of 40 animals in each group. E. granulosus infection was modeled in mice by intraperitoneal injection of 10 000 protoscoleces per mouse. Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) and peripheral interleukin-4 (IL-4) and IL-10 levels were measured 2, 8, 30, 60, 180 days post-infection. Mouse liver specimens were excised for hematoxylin-eosin (HE) staining and immunostaining, and ICOS expression was quantified in mouse liver specimens using quantitative real-time PCR (qPCR) assay. Results There were no significant differences in serum ALT (F = 12.082, P < 0.05), AST (F = 6.347, P < 0.05) or ALP levels (F = 52.186, P < 0.05) in mice 2, 8, 30, 60 and 180 days post-infection with E. granulosus. The serum ALT levels were significantly higher in the infection group than in the control group 2 [(61.72 ± 9.89) vs. (50.65 ± 4.67)U/L, P < 0.05] and 30 days post-infection [(80.61 ± 23.71)vs.(67.75 ± 9.79)U/L, P < 0.05], and the serum ALT levels were significantly higher in the infection group than in the control group 2 [(181.06 ± 60.61) vs.(115.58 ± 17.66)U/L, P < 0.05] and 180 days post-infection [(137.84 ± 29.01) vs. (108.05 ± 10.33) U/L, P < 0.05], while greater serum ALP levels were measured in the infection group than in the control group 2 [(162.90 ± 21.04)vs.(64.54 ± 5.99)U/L, P < 0.05], 8[(176.36 ± 24.56) vs. (62.70 ± 9.21)U/L, P < 0.05] and 30 days post-infection [(138.86 ± 13.59) vs. (58.60 ± 5.28) U/L, P < 0.05]. A few inflammatory cells were seen in mouse liver in the infection group 30 days post-infection, and no apparent changes were found in the mouse hepatic structure 60 days post-infection. On day 180 post-infection, a large number of epithelium-like cells presented fibrotic growth in mouse liver in the cyst-infiltrating regions, with cuticula formation seen, and plenty of red cells were present in lesions and hepatocyte space. Positive ICOS expression was detected in mouse liver in the infection group, with ICOS-positive cells predominantly seen in the cytoplasm of the hepatocyte, and the ICOS expression increased over time. The relative ICOS mRNA expression was 2.732 ± 0.094 on day 180 post-infection, which was significantly greater than that on day 2 postinfection (0.746 ± 0.049). There were no significant differences in serum IL-4 or IL-10 levels at different time points after E. granulosus infections, while the serum IL-4 and IL-10 levels peaked in the infection group 180 days and 60 days post-infection, respectively. Higher serum IL-4 levels were measured in the infection group than in the control group 8 [(22.50 ± 3.24) vs. (5.82 ± 0.49) pg/mL, P < 0.05], 30 [(15.49 ± 4.73) vs. (5.10 ± 1.38) pg/mL, P < 0.05], 60 [(36.93 ± 6.14) vs. (4.13 ± 1.19) pg/mL, P < 0.05] and 180 days post-infection [(198.35 ± 0.70) vs. (4.19 ± 0.98) pg/mL, P < 0.05], and higher IL-10 levels were measured in the infection group than in the control group 2 [(4.84 ± 1.91) vs. (2.11 ± 1.03) pg/mL, P < 0.05], 8 [(44.72 ± 14.63) vs. (3.16 ± 0.60) pg/mL, P < 0.05], 30 [(25.47 ± 8.00) vs. (3.83 ± 1.87) pg/mL, P < 0.05], 60 [(187.16 ± 60.44) vs. (3.69 ± 1.05) pg/mL, P < 0.05] and 180 days post-infection [(85.40 ± 7.15) vs. (3.25 ± 0.93) pg/mL, P < 0.05]. Conclusions High ICOS expression is present in the liver of mice with E. granulosus infections. The positive ICOS expression and immune activation levels increase with the time of E. granulosus infections, leading to aggravation of hepatocyte injury caused by inflammation.
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Micheliolide is a kind of guaiacane sesquiterpene lactone from natural plants. It distributs in the root bark of Cephalantheropsis gracilis and Michelia compressa . Modern pharmacological studies show that ，micheliolide can play an anti-tumor role by inhibiting the proliferation of tumor cells ，regulating the apoptosis of tumor cells ，inducing autophagy of tumor cells ，and inhibiting invasion and metastasis of tumor cells. It can reduce the expression of inflammatory cytokines by regulating several related signaling pathways ，such as nuclear factor κB and phosphatidylinositol-3-hydroxykinase/protein kinase B ；it can play an immunomodulatory role by restoring the expression of immune-related factors in the body ；it can play a neuroprotective effect by reducing the accumulation of amyloid β-protein and inhibiting microglial activation ；it can play a liver protective role by reducing the inflammatory response and steatosis of hepatocytes ；its nephroprotective mechanism is related to the relief of the inflammatory response by regulating multiple pathways ；the mechanisms of its cardioprotective action include alleviating the inflammatory response and reducing the level of oxidative stress ，etc. In conclusion ，micheliolide is a compound of great development value.
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Objective:To investigate the role of chemokine receptor CX3CR1 in chronic skin inflammation and its regulatory mechanism.Methods:Wild type (WT) C57BL/6 mice and Cx3 cr1 GFP/GFP mice were induced by DNFB to establish acute and chronic allergic contact dermatitis (ACD) model. Ear inflammation and swelling were observed with hematoxylin-eosin (HE) staining. Flow cytometry (FCM) was used to detect the changes in classical Langerhans cell (LC) and monocyte-derived LC (Mo-LC), as well as the expression of major histocompatibility complex Ⅱ (MHCⅡ), inducible nitric oxide synthase (iNOS) and TNF-α. Changes in epidermal LC in UV irradiation-induced dermatitis models were also analyzed. In human chronic skin inflammation, CX3CL1 expression was detected using immunohistochemistry, RT-PCR and Western blot and CD1a, CD14 and CD207 expression was observed with immunofluorescence staining. Results:In the chronic ACD model, Cx3 cr1 GFP/GFP mice showed significantly alleviated ear inflammatory and swelling as compared with WT mice, but no significant difference was found in the acute ACD model. The percentages of Mo-LC were decreased in the chronic ACD model and after three weeks of UV irradiation. Moreover, MHCⅡ, TNF-α and iNOS expressed by Mo-LC were significantly upregulated as compared with those by classical LC. CX3CL1 expression was significantly upregulated and the numbers of CD14 + monocytes and CD1a + langerin - Mo-LC were dramatically increased in human chronic skin inflammation. Conclusions:CX3CR1 might maintain inflammatory response by regulating local remodeling of Mo-LC in chronic skin inflammation.
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ObjectiveTo investigate the efficacy of Bushen Shengxue prescription and Yiqi Yangxue prescription in the treatment of chronic aplastic anemia and the effect on T cell subsets and the expression of T-box expressed in T cells （T-bet） and GATA binding protein 3 （GATA3）. MethodA total of 585 patients with chronic aplastic anemia who were treated in 19 hospitals in China from May 2018 to June 2021 were enrolled. With the prospective， double-blind and randomized control methods， the patients were randomized into three groups： kidney deficiency group， Qi and blood deficiency group， and control group. The three groups were respectively treated with Bushen Shengxue prescription granule， Yiqi Yangxue prescription granule， and Placebo （half the dose of Bushen Shengxue formula granules）. In addition， all of them were given oral cyclosporin and androgen. The treatment lasted 6 months， with 3 months as a course. The blood routine indexes， T cell subsets， and fusion genes T-bet and GATA3 before and after treatment were analyzed， and the safety indexes were monitored. ResultDuring the observation， a total of 75 cases dropped out and 18 were rejected. Finally， 161 cases in the kidney deficiency group， 164 in the Qi and blood deficiency group， and 167 in the control group were included. After 6 months of treatment， the total effective rate was 98.8% （159/161） in the kidney deficiency group， which was higher than the 79.9% （131/164） in the Qi and blood deficiency group （χ2=30.135， P<0.01） and the 61.7% （103/167） in the control group （χ2=70.126， P<0.01）. The total effective rate was higher in the Qi and blood deficiency group than in the control group （χ2=13.232， P<0.01）. After treatment， the hemoglobin （HGB） content increased significantly in three groups （P<0.05） as compared with that before treatment， particularly the kidney deficiency group （P<0.01）. After treatment， the white blood cell （WBC） count and platelet （PLT） count in the kidney deficiency group and the control group increased compared with those in the Qi and blood deficiency group （P<0.01）. There was no specific difference in neutrophils （ANC） after treatment among the three groups. At the same time point， the level of T helper type 1 （Th1） cells， Th1/Th2 ratio （P<0.05）， level of CD4+， and CD4+/CD8+ ratio （P<0.05） were significantly low in the kidney deficiency group among three groups. There was no significant difference in CD19-， HLA/DR+， and CD25+ between the kidney deficiency group and the other two groups， but the T-bet of the kidney deficiency group and the control group was lower than that of the Qi and blood deficiency group （P<0.05）. ConclusionBushen Shengxue prescription exerts therapeutic effect on the aplastic anemia by improving the immunoregulatory mechanism， inhibiting the activity of immune system， modulating T cell subsets， suppressing Th1 and CD4+， and promoting bone marrow hematopoiesis. Moreover， it is safe with little side effects， which is worthy of further promotion.
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ObjectiveTo explore the pharmacodynamic ingredients of Zhenqi Fuzheng granules （ZFG） for immunomodulatory through spectrum-effect relationship analysis， which provides experimental basis for improving the quality standard of ZFG. MethodEighteen batches of ZFG from six manufacturers were collected for analysis. The fingerprints were established by high performance liquid chromatography （HPLC）. Acetonitrile （A）-0.1% formic acid aqueous solution （B） were adopted as the mobile phase with gradient elution （0-15 min， 5%A； 15-23 min， 5%-8%A； 23-30 min， 8%-11%A； 30-45 min， 11%-18%A； 45-60 min， 18%-21%A； 60-67 min， 21%-23%A； 67-90 min， 23%-37%A）， the detection wavelength was 220 nm. Chemometric analysis such as similarity analysis and hierarchical cluster analysis （HCA） were subsequently used to analyze the similarities and chemical differences among these samples. A cyclophosphamide-induced immunodeficiency mouse model was used to evaluate the immune-enhancing effects of the products from different manufacturers. The spectrum-effect relationship between HPLC fingerprints and the immunomodulatory effects was examined using Spearman bivariate correlation analysis. HPLC coupled with mass spectrometry （HPLC-MSn） was used to identify the spectrum-effect related peaks with electrospray ionization， positive and negative ion modes， and scanning range of m/z 100-1 500. ResultThe HPLC fingerprint of ZFG was established， and twenty peaks with good resolution were selected as common peaks. The results of quality analysis and pharmacodynamic test showed there were significant differences in both ingredients content and immune-enhancing effects of ZFG from different manufacturers. Through spectrum-effect relationship study， twelve peaks were screened as bioactive ingredients peaks. Thereafter， eight peaks among them were subsequently identified by HPLC-MSn. They were salidroside （peak 2）， echinacoside （peak 5）， calycosin-7-glucoside （peak 6）， isomer of specnuezhenide （peak 7）， isonuezhenide （peak 9）， calycosin （peak 11）， nuezhenide G13 or oleonuezhenide （peak 14）， and formononetin （peak 18）， respectively. ConclusionThere are differences in quality and efficacy of ZFG produced by different manufacturers. Through spectrum-effect relationship analysis， the medicinal ingredients of ZFG for immune-enhancing effects are screened， which can provide reference for the improvement of its quality standard.
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ObjectiveTo determine the therapeutic effect of Gegentang granules on a disease-syndrome mouse model combining human coronavirus 229E （hCoV-229E） pneumonia with Hanshi Yidu Xifei syndrome in vivo. MethodMice were randomly divided into normal group， infection group， cold-dampness group， model group， chloroquine phosphate group （0.18 g·kg-1）， interferon-α2b （IFN-α2b） group （1.83×106 U·kg-1）， Gegentang granules high-dose and low-dose groups （6.6， 3.3 g·kg-1） with 10 mice in each group. Cold-dampness environment and hCoV-229E infection were used for modeling， and the general status and lung index of mice in each group were observed. The viral load in lung tissue was detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， the pathological changes in lung tissue were evaluated by hematoxylin-eosin （HE） staining， the levels of serum gastrointestinal hormones and inflammatory factors in lung tissue were detected by enzyme-linked immunosorbent assay （ELISA）， and the percentage of peripheral blood lymphocytes was detected by flow cytometry. ResultComparing with model group， Gegentang granules could significantly alleviate the physical signs of Hanshi Yidu Xifei syndrome， including listlessness， weakness of limbs， sticky stool， etc. Comparing with model group， Gegentang granules high-dose group significantly reduced lung index， histopathological score of interstitial lung and bronchus， and the level of serum motilin （P<0.05， P<0.01）， two doses of Gegentang granules could significantly increase the level of serum gastrin （P<0.05， P<0.01）， the percentage of CD4+， CD8+ T lymphocytes in peripheral blood （P<0.05， P<0.01）， and the level of tumor necrosis factor-α （TNF-α） in lung tissue was significantly decreased （P<0.01）， and the level of interleukin-6 （IL-6） showed decreasing tendency. ConclusionGegentang granules has therapeutic effect on model mice. It can improve the appearance and behavior characterization， regulate the level of gastrointestinal hormones， decrease lung index and histopathological score， and possibly play an immunomodulatory role by inhibiting the expression of inflammatory cytokines in lung tissue and restoring the percentage of peripheral blood lymphocytes.
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ObjectiveTo determine the therapeutic effect of Gegentang granules on a disease-syndrome mouse model combining human coronavirus 229E （hCoV-229E） pneumonia with Hanshi Yidu Xifei syndrome in vivo. MethodMice were randomly divided into normal group， infection group， cold-dampness group， model group， chloroquine phosphate group （0.18 g·kg-1）， interferon-α2b （IFN-α2b） group （1.83×106 U·kg-1）， Gegentang granules high-dose and low-dose groups （6.6， 3.3 g·kg-1） with 10 mice in each group. Cold-dampness environment and hCoV-229E infection were used for modeling， and the general status and lung index of mice in each group were observed. The viral load in lung tissue was detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， the pathological changes in lung tissue were evaluated by hematoxylin-eosin （HE） staining， the levels of serum gastrointestinal hormones and inflammatory factors in lung tissue were detected by enzyme-linked immunosorbent assay （ELISA）， and the percentage of peripheral blood lymphocytes was detected by flow cytometry. ResultComparing with model group， Gegentang granules could significantly alleviate the physical signs of Hanshi Yidu Xifei syndrome， including listlessness， weakness of limbs， sticky stool， etc. Comparing with model group， Gegentang granules high-dose group significantly reduced lung index， histopathological score of interstitial lung and bronchus， and the level of serum motilin （P<0.05， P<0.01）， two doses of Gegentang granules could significantly increase the level of serum gastrin （P<0.05， P<0.01）， the percentage of CD4+， CD8+ T lymphocytes in peripheral blood （P<0.05， P<0.01）， and the level of tumor necrosis factor-α （TNF-α） in lung tissue was significantly decreased （P<0.01）， and the level of interleukin-6 （IL-6） showed decreasing tendency. ConclusionGegentang granules has therapeutic effect on model mice. It can improve the appearance and behavior characterization， regulate the level of gastrointestinal hormones， decrease lung index and histopathological score， and possibly play an immunomodulatory role by inhibiting the expression of inflammatory cytokines in lung tissue and restoring the percentage of peripheral blood lymphocytes.