BACKGROUND:Immunosuppression leads to impaired body immune function and aggravates the disease.Herb-partitioned moxibustion can effectively regulate immune function and improve immunity in the body,but its regulatory mechanism has not been elucidated. OBJECTIVE:To sequence immunosuppressed model rats treated with herb-partitioned moxibustion using bioinformatics techniques based on transcriptomics and to explore the mechanisms by which it regulates immunity. METHODS:Twenty-four Sprague-Dawley rats were randomly assigned to three groups:control,model,and herb-partitioned moxibustion groups,with eight rats in each group.The model and herb-partitioned moxibustion groups were subjected to establishment of an immune suppression model by intraperitoneal injection of cyclophosphamide at a dose of 35 mg/kg for 3 consecutive days.No interventions were administered to the control and model groups after modeling.In contrast,the herb-partitioned moxibustion group received moxibustion treatment at Zhongwan,Shenque,Guanyuan,and Zusanli acupoints using a combination of moxa and herbal cakes,once a day,for 10 consecutive days,with samples being collected the day after the end of the intervention.Peripheral blood was collected from all groups of rats to measure their white blood cell count.RNA-seq was performed on the Illumina sequencing platform,and differentially expressed genes were selected for bioinformatics analysis using the GO and KEGG databases. RESULTS AND CONCLUSION:Compared with the control group,the model group exhibited a significant decrease in white blood cell count(P<0.001).RNA-seq analysis identified 3 026 differentially expressed genes between the model and control groups,with 1 565 upregulated and 1 461 downregulated.There were 535 differentially expressed genes identified between the herb-partitioned moxibustion group and the model group,with 280 upregulated and 255 downregulated.The Venn diagram analysis revealed that 159 genes were downregulated in the model group compared with the control group.However,after moxibustion with herbal cakes,these genes were upregulated.Protein-protein interaction network analysis identified 10 core targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,Mx1,Syk,Hspa1a,and Ret.According to GO and KEGG analyses,moxibustion with herbal cakes regulated the body through pathways related to immune response,viruses,angiogenesis,and the autoimmune system.To conclude,there is a significant association between herbal cake-separated moxibustion intervention and immune suppression targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,and Mx1.The intervention exhibits regulatory effects in the pathways related to immune responses,viral activities,and angiogenesis.

Objective:To compare similarities and differences in expression changes of cytotoxic T-lymphocyte antigen-4(CT-LA-4)and programmed death receptor 1(PD-1)in immunosuppressed rabbits under different moxibustion interventions.Methods:Twenty large-eared white rabbits were randomly divided into normal group,model group,moxa stick moxibustion(MSM)group and herbal cake-partitioned moxibustion(HPM)group,with five rabbits in each group.CTX-induced immunosuppressed models were prepared by intraperitoneal injection for 7 consecutive days.After successful modeling,MSM and HPM were performed on alternate days for 10 treatments.Rabbits were anesthetized after treatment,and serum,liver and spleen were collected.Serum PD-1 and PD-L1 contents were detected by ELISA.PD-1 in liver tissue was detected by immunohistochemistry,and CTLA-4 mRNA in liver and spleen tissues were detected by RT-qPCR.Results:Both HPM and MSM could reduce PD-1 and PD-L1 levels due to immunosuppression,and could effectively suppress elevated levels of CTLA-4 in spleen and PD-1 and CTLA-4 in liver,which were statistically different compared with immunosuppression model group(P<0.05).Pearson correlation test showed that CTLA-4 and PD-1 in liver tissue had significant positive correlation(r=0.780 7,P<0.001).Conclusion:HPM can improve body immune function by regulating multiple immunosuppressive sites.

Objective:To observer the effect of electroacupuncture at "Neiguan", "Ximen" and "Hegu" acupoints of the Pericardium Meridian and observe the expression of CaMK Ⅱ mRNA and apoptosis in the myocardial ischemic rats with reperfused injury.Methods:The Wister rats were randomly divided into sham operation group, model group, Acupuncture Neiguan group, Acupuncture Ximen group and Acupuncture Hegu group, with 10 rats in each group. Exceptthe sham operation group, the othergroups were all ligated with the left anterior descending branch of the coronary artery. Before ligating the coronary artery, electroacupuncture was performed at "Neiguan", "Ximen" and "Hegu" points for 20 minutes respectively. After the ligation for 40 minutes, electroacupuncture was performed at the above points for 20 minutes and then restore coronary perfusion for 60 minutes. The ECG changes of rats were recorded. The level of CaMK Ⅱ mRNA was detected by RT-PCR and the apoptosis rate of cardiomyocytes was detected by TUNEL. The correlation between the expression rate of CaMK Ⅱ mRNA and the apoptosis rate of cardiomyocytes was analyzed according to the pearson correlation.Results:At 20, 40 and 60 min after the ligation of coronary artery and 30 and 60 min after the loosening of coronary artery, compared with the model group and Acupuncture Hegu group, the ST segment difference of Acupuncture Neiguan group and Acupuncture Ximen group was decreased ( P<0.01); The levels of CaMK Ⅱ mRNA (0.483 ± 0.050, 0.432 ± 0.079 vs. 0.935 ± 0.109) in Acupuncture Neiguan group and Acupuncture Ximen group were significantly decreased ( P<0.01). The apoptosis rate of cardiomyocytes (10.86% ± 2.17%, 9.66% ± 4.09% vs. 36.22% ± 1.69%) significantly decreased ( P<0.01). Conclusions:There is a positive correlation between CaMK Ⅱ mRNA level and cardiomyocyte apoptosis during myocardial ischemia-reperfusion. Acupuncture can effectively reduce CaMK Ⅱ mRNA level and cardiomyocyte apoptosis rate and protect cardiomyocytes.

Objective To explore the effect of polysaccharides from Sargassum graminifolium (SGP) on the intestinal flora of obese mice and the relationship between changes of the intestinal flora and biochemical indicators of blood lipid.Methods Sixty c57BL/6J mice were randomly divided into the conventional feed group (the control group,n =10) and the high-fat feed group (n =50).The model of obesity was established by using the animals in the high-fat feed group.With rejection of 10 animals that were not conformed to the selection standards,the remaining 40 mice were divided into the blank control group and the treatment group.The treatment group was subdivided into the low-dosage group that was given SGP at a dosage of 50 mg/kg by gavage,the medium-dosage group that was given SGP at a dosage of 100 mg/kg by gavage,and the high-dosage group that was given SGP at a dosage of 200 mg/kg by gavage,each consisting of 10 animals.After 8 weeks of feeding the SGP by gavage,detections were performed in biochemical indicators of blood lipid and the number of colonies of Escherichia coli,lactobacillus and bifidobacterium in the contents of the appendix.Results The number of colonies of lactobacillus [(7.05 ± 0.15)lg CFU/g] and that of bifidobacterium [(8.75 ± 0.41)lg CFU/g] in the blank control group was significantly decreased,as compared with that of the control group [(8.31 ± 0.12)lg CFU/g and(10.17 ± 0.31) lg CFU/g].The number of colonies of Escherichia coli in the blank control group [(6.87± 0.25)lg CFU/g] was significantly higher,as compared with that of the control group [(5.83 ± 0.28) lg CFU/g],and statistical significance could be seen,when comparisons were made between them(P ＜ 0.05).When compared with that of the blank control group,the number of colonies of Escherichia coli in the medium-dosage and high-dosage groups were significantly decreased.The number of colonies of lactobacillus and bifidobacterium in the low-dosage,medium-dosage and high-dosage groups were significantly increased.The number of colonies of bifidobacterium in the low-dosage and high-dosage groups were also significantly increased,with statistical significance(P ＜0.05).TC [(5.18 ± 0.87) mmol/L],TG [(1.90 ± 0.21) mmol/L] and LDL [(0.63 ± 0.02) mmol/L] were all higher than those of the control group,but were lower than those in the blank group,which were respectively [TC (2.54 ± 0.39) mmol/L],[TG (1.12 ± 0.20) mmol/L] and [LDL(0.35 ±0.05)mmol/L].As compared with those of the blank control group,the levels of HDL in the low-dosage,medium-dosage and high-dosage groups were all decreased significantly.The levels of TG in the medium-dosage and high-dosage groups were also decreased significantly,the levels of LDL in the low-dosage,medium-dosage and high-dosage groups were all decreased significantly,and the levels of HDL in the mediumdosage and high-dosage groups were all increased significantly.Statistical significance could be noticed,when comparisons were made between them(P ＜ 0.05).The levels of Escherichia coli were positively correlated with those of TC,TG and LDL,while the levels of HDL coli were negatively correlated with those of TC,TG and LDL.The levels of lactobacillus and bifidobacterium were negatively correlated with those of TC,TG and LDL,however,they were positively correlated with those of HDL.Conclusions SGP could suppress the growth of Escherichia coli in the intestinal of the high-fat-feed mice,and the number of intestinal colonies was correlated with the biochemical indicators of blood lipid.

Objective To explore the effect of polysaccharides from Sargassum graminifolium (SGP) on the intestinal flora of obese mice and the relationship between changes of the intestinal flora and biochemical indicators of blood lipid.Methods Sixty c57BL/6J mice were randomly divided into the conventional feed group (the control group,n =10) and the high-fat feed group (n =50).The model of obesity was established by using the animals in the high-fat feed group.With rejection of 10 animals that were not conformed to the selection standards,the remaining 40 mice were divided into the blank control group and the treatment group.The treatment group was subdivided into the low-dosage group that was given SGP at a dosage of 50 mg/kg by gavage,the medium-dosage group that was given SGP at a dosage of 100 mg/kg by gavage,and the high-dosage group that was given SGP at a dosage of 200 mg/kg by gavage,each consisting of 10 animals.After 8 weeks of feeding the SGP by gavage,detections were performed in biochemical indicators of blood lipid and the number of colonies of Escherichia coli,lactobacillus and bifidobacterium in the contents of the appendix.Results The number of colonies of lactobacillus [(7.05 ± 0.15)lg CFU/g] and that of bifidobacterium [(8.75 ± 0.41)lg CFU/g] in the blank control group was significantly decreased,as compared with that of the control group [(8.31 ± 0.12)lg CFU/g and(10.17 ± 0.31) lg CFU/g].The number of colonies of Escherichia coli in the blank control group [(6.87± 0.25)lg CFU/g] was significantly higher,as compared with that of the control group [(5.83 ± 0.28) lg CFU/g],and statistical significance could be seen,when comparisons were made between them(P ＜ 0.05).When compared with that of the blank control group,the number of colonies of Escherichia coli in the medium-dosage and high-dosage groups were significantly decreased.The number of colonies of lactobacillus and bifidobacterium in the low-dosage,medium-dosage and high-dosage groups were significantly increased.The number of colonies of bifidobacterium in the low-dosage and high-dosage groups were also significantly increased,with statistical significance(P ＜0.05).TC [(5.18 ± 0.87) mmol/L],TG [(1.90 ± 0.21) mmol/L] and LDL [(0.63 ± 0.02) mmol/L] were all higher than those of the control group,but were lower than those in the blank group,which were respectively [TC (2.54 ± 0.39) mmol/L],[TG (1.12 ± 0.20) mmol/L] and [LDL(0.35 ±0.05)mmol/L].As compared with those of the blank control group,the levels of HDL in the low-dosage,medium-dosage and high-dosage groups were all decreased significantly.The levels of TG in the medium-dosage and high-dosage groups were also decreased significantly,the levels of LDL in the low-dosage,medium-dosage and high-dosage groups were all decreased significantly,and the levels of HDL in the mediumdosage and high-dosage groups were all increased significantly.Statistical significance could be noticed,when comparisons were made between them(P ＜ 0.05).The levels of Escherichia coli were positively correlated with those of TC,TG and LDL,while the levels of HDL coli were negatively correlated with those of TC,TG and LDL.The levels of lactobacillus and bifidobacterium were negatively correlated with those of TC,TG and LDL,however,they were positively correlated with those of HDL.Conclusions SGP could suppress the growth of Escherichia coli in the intestinal of the high-fat-feed mice,and the number of intestinal colonies was correlated with the biochemical indicators of blood lipid.