Objective To investigate the different immune status of TCM classification and the levels of T and B cells and the re-lationship between the NK cells .Methods Three different immune state of patients with chronic HBV were divided into asthenia syndrome(AS) group and sthenia syndrome(SS) group ,and then we detected the peripheral blood lymphocyte subsets and NK cells ,and compared the above each index analysis .Results In three different immune status ,all patients with AS and SS groups CD4+ T cell percentages are lower than normal control group (P<0 .05) ,and in a state of immune tolerance ,CD4+ T cells of SS group were lower than the AS group (P<0 .05) ,in a state of immune clearance ,CD8+ T cell percentage were SS group> control group> the AS group (P<0 .05) .Three different immune status ,the ratio of CD4+ /CD8+ SS group were significantly lower than the AS group or control group ,in a state of immune clearance the SS and the AS group B lymphocyte percentage were higher than the control group ,the percentage of NK cells was lower than the control group (P<0 .05) .Conclusion Chronic HBV infection have immune dysfunction ;HBV infected different immune status of TCM classification have relationship with T and B cell subsets and NK cells ,and the immune function test has certain clinical application value for the judgment of TCM syndrome type .

Objective:To investigate the expression level of C-type lectin domain family 4 member G ( CLEC4 G) in liver disease tissues and its correlation with the clinicopathological characteristics of hepatocellular carcinoma (HCC) patients. Methods:The cancer tissue and the corresponding adjacent tissues (at least 2 cm from the edge of the cancer tissue), cut in surgeries from January to December in 2019, of 40 HCC patients in Zhuzhou Central Hospital, as well as 10 normal liver tissue samples (seen as far away as possible from the edge of the cancer tissue with naked eyes) and 10 liver cirrhosis samples were analyzed retrospectively. The tumor genome atlas (TCGA) database was used to screen the HCC transcriptome data sets, and bioinformatics methods were used to make expression heat maps and box maps which can help analyze the difference of CLEC4 G in cancer and adjacent tissues. The mRNA expression level of CLEC4 G was detected by conducting real-time fluorescence quantitative PCR (qRT-PCR), and the protein expression level of CLEC4G was detected by immunohistochemistry (IHC). The measurement data were expressed as mean±standard deviation ( Mean± SD). Group t test was used for inter-group comparison. The counting information was expressed as a percentage (%). The χ2 test was adopted to analyze the correlation between CLEC4 G expression level and the clinicopathological features of patients. Results:The expression level of CLEC4 G in cancer tissues was significantly decreased in heat map compared with that in adjacent tissues. In the box figure, the relative expression of CLEC4 G mRNA in the cancer tissues was (82.5±18.9) and (3 354.4±296.2) in paracancer tissues, with statistically significant difference ( P<0.001). Respectively, qRT-PCR and IHC showed that mRNA of CLEC4 G were abundant in normal liver tissues (3 301.3±286.4), while they were very little in liver cancer tissues (63.6±32.9), significantly decreasing in liver cirrhosis (1 742.6±208.7) and paracancer tissues (1 553.2±249.9), with statistically significant difference ( P<0.001). Moreover, low CLEC4 G expression level was associated with tumor vascular metastasis in HCC patients. Conclusions:CLEC4 G is highly expressed in normal liver tissue, but with the progression of malignant liver disease, it is significantly decreased with little expression in HCC tissue. It can be expected to be a good marker for the pathological diagnosis of HCC.

Objective: To investigate the regulation of emodin on microRNA expressions in mouse model with sepsis by GeneChip microRNA array. Methods: Forty two c57 mice were randomly (random number) divided into 3 groups: sham operation group (sham group, n=14),sepsis group(n=14) and emodin group (n=14). The sepsis model of the mouse was subjected to cecal ligation and puncture (CLP). Mice in emodin group received intraperitoneal injection of emodin (40 mg/kg) half an hour before operation and every 12 hours after CLP. All mice were sacrificed 48 h after surgery and part of the ileum were removed for intestine tissue stained with hematoxylin eosin. The levels of tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6) and intestinal fatty acid binding protein (I-FABP) in peripheral blood were measured by enzyme-linked immunosorbent assay (ELISA). Total RNA of ileum tissues was extracted from the sepsis group and emodin group, and then subjected to miRNA microarray. The results of microarray were further verified by quantitative real-time PCR (qRT-PCR). Target genes of differentially expressed miRNAs were predicted and subjected to gene ontology (GO) and KEGG pathway enrichment analysis. Data of multi-groups were analyzed by one way variance (ANOVA) and inter-group comparisons were made by SNK-q tests. Mann-Whitney U test was used when homogeneity of variance were not met. The value of P<0.05 was considered statistically significant. Results: Compared to the sepsis group, the levels of serum TNF-α, IL-6 and I-FABP in the emodin group were decreased significantly. MiRNA microarray showed that 23 miRNAs were differentially expressed in the emodin group as compared to the sepsis group, among which 17 miRNAs were up-regulated and 6 miRNAs were down-regulated. qRT-PCR results were consistent with miRNA array data. Target genes of 10 selected miRNAs were predicted and subjected to bioinformatic analysis. A total of 3 410 target genes were significantly enriched in 2 072 GO biological process terms, 246 GO cellular component items and 277 GO molecular function terms. Moreover, KEGG pathway analysis showed that these differential miRNAs were involved in the regulation of PI3K-Akt signaling pathway, MAPK signaling pathway, and TNF signaling pathway. Conclusions Emodin can regulate the expression of multiple microRNAs, and play an important role in protecting the intestinal mucosal barrier via a variety of targets and pathways.