Objective To study the effects of Ganoderma lucidum polysaccharide (GLP) on PBMCs and the related immune mechanism.Methods PBMCs from cancer patients and healthy donors were isolated and treated with different doses of Ganoderma lucidum polysaccharides (10 ng/ml,50 ng/ml and 100 ng/ml).DC cell costimulatory molecules (HLA-DR,CD83,and CD11 c),Th1 (CD3 + CD8-IFN-γ+) cells,Th2 (CD3 + CD8-IL-4+) cells and NK (CD3-CD56+) were analyzed by FCM.Furthermore,The CD3+ CD4+ Th ceils were separated by immunomagnetic beads and stimulated with Ganoderma lucidum polysaccharides at different concentrations in culture.After 24 h,the cytokine expression levels of Th1 and Th2were detected by RT-PCR.The expressions of Th1 differentiation-related transcription factor,STAT4,were analyzed by Western blot.Results Ganoderma lucidum polysaccharides can significantly stimulate in vitro Thl cell differentiation (P＜0.01) in a dose depend manner.It correlates with an increased expression of STAT4 and the elevated mRNA expression levels of Th1 cytokineincluding IL-12,IFN-γ and TNF-α (P＜0.01).Conclusion Ganoderma lucidum polysaccharides may promote Th1 differentiation and increase the secretion of Th1 cvtokines through the upregulation of STAT4.

Objective To investigate the regulation of the immune function of cytokine-induced killer cells(CIK) by dendritic cell (DC) vaccine in the patients with chronic hepatitis B(CHB) in vitro.Methods CIK cells from 30 patients with CHB were cultured in vitro,and were randomly divided into two groups,DC vaccine-treated group and the control group.After 14 days of culture,the percentages of CD3 + CD4+ T,CD3 +CD8 +T and CD3 +CD56+ T cells among CIKs were analyzed by flow cytometry.The levels of IL-12,IFN-γand IL-6 in cell culture supernatant was detected by ELISA.Results The percentages of CD3 + CD4 + T,CD3 +CD8+T and CD3+ CD56+ T cells were 18.27％,64.36％ and 20.00％ in CIKs in DC vaccine group,and 17.79％ ( P ＞ 0.05 ),54.69％ ( P ＜ 0.01 ) and 13.39％ ( P ＜ 0.01 ) in the control group,respectively.The perentage of CD3 + CD4 + T cells were similar between the two groups ( P ＞ 0.05 ),but for the perentage of CD3 + CD8 +T and C D3 + CD56 + T cells were significantly different between the two groups (t =4.130 and 5.601respectively,Ps ＜ 0.01 ).The concentrations of IL- 12,IL-4 and IFN-γin supernatant were ( 177.82 ± 130.06),(31.77 ± 9.52) and (86.99 ± 56.30) ng/L in DC vaccine-treated group respectively,which were significantly different from those of (80.83 ±50.15) ng/L (t =3.811,P ＜0.01 ),(40.33 ± 19.74) ng/L( t =2.141,P ＜0.05) and (42.07 ± 19.68)ng/L(t =4.125,P ＜0.01) in the control group,respectively.Conclusion DC vaccine could enhance the killing function of CIK cells.

The core protein (CP) of the classical swine fever virus (CSFV) is one of its structural proteins. Apart from forming the nucleocapsid to protect internal viral genomic RNA, this protein is involved in transcriptional regulation. Also, during viral infection, the CP is involved in interactions with many host proteins. In this review, we combine study of this protein with its disorders, structural/functional characteristics, as well as its interactions with the non-structural proteins NS3, NS5B and host proteins such as SUMO-1, UBC9, OS9 and IQGAP1. We also summarize the important part played by the CP in CSFV pathogenicity, virulence and replication of genomic RNA. We also provide guidelines for further studies in the CP of the CSFV.
Animals ; Classical Swine Fever ; virology ; Classical swine fever virus ; genetics ; metabolism ; pathogenicity ; Genome, Viral ; Swine ; Viral Core Proteins ; chemistry ; genetics ; metabolism ; Virulence

AIM:To investigate the effects of glucagon-like peptide-1 (GLP-1) on mRNA expression of SOCS-3 and SREBP-1c in the rats with nonalcoholic fatty liver disease.METHODS:Male SD rats were randomly divided into normal control ( NC) group, high fat ( HF) group and HF+liraglutide ( Lira) group.The rats in HF group and HF+Lira group were given high-fat diet for 16 weeks.After 12 weeks of high-fat diet feeding in HF+Lira group, Lira (600μg? kg-1? d-1 ) was intraperitoneally injected for 4 weeks.At the end of the 16th week, the rats were killed.The pathologi-cal changes of the liver were observed under optical microscope.The serum levels of alanine aminotransferase ( ALT) , as-partate aminotransferase ( AST) , triglyceride ( TG) and total cholesterol ( TC) were detected by automatic biochemical an-alyzer.TG contents of liver were measured by GPO-PAP method.The fasting insulin ( FINS) was determined by ELISA, and insulin resistance index was assessed by homeostasis mode assessment ( HOMA-IR) .The mRNA expression of SOCS-3 and SREBP-1c in the liver tissues was detected by RT-qPCR.RESULTS:Compared with NC group, HOMA-IR, TG of liver, and the serum levels of ALT, AST, TG, TC and FINS in HF group were obviously increased (P<0.01).Compared with HF group, HOMA-IR, TG of liver, and the serum levels of ALT, AST, TG, TC and FINS in HF+Lira group were all obviously decreased (P<0.05 or P<0.01).The mRNA expression of SOCS-3 and SREBP-1c in HF group was signifi-cantly higher than that in NC group (P<0.01).The mRNA expression of SOCSV3 and SREBP-1c in HF+Lira group was significantly decreased as compared with HF group (P<0.05).CONCLUSION:Liraglutide may improve the IR and re-
duce TG of liver through decreasing the mRNA expression of SOCS-3 and SREBP-1c, so as to play a therapeutic role in nonalcoholic fatty liver disease.

Objective To observe expression and location of cannabinoid receptor 1 (CB1) in liver tissue of patients with chronic hepatitis B (CHB) ,and analyze the relationship of it with the liver fibrosis score,the serum levels of TGF-β1 and Leptin. Methods Liver biopsies were performed in 118 patients with CHB.The expression of CB1 in liver tissue was observed by immune histochemical staining, and semi-quantitative analysis was carried out to devide the CB1 score into four grades: -, +, + +, + + +. Serum levels of TGF-β1 and Leptin were determined by ABC-ELISA double-antibody sandwich method. Results The expression of CB1 in liver tissue with CHB had significant relationship with the fibrosis score. As the expression of the CB1 increased, the fibrosis score became higher ( F = 23. 369,P = 0. 000). Moreover, the expression of CB1 in liver tissue with CHB had significant relationship with the serum levels of TGF-β1 and Leptin( F values were 8. 762 and 5. 749;P values were 0. 001 and 0. 027, respectively). Conclusion CB1 may play promotive role in the process of hepatic fibrosis through regulation of TGF-β1 and Leptin.

The architecture of the alveolus and its capillaries of human lung injected withsolution of ABS in methyl ethyl ketone was studied under the SEM.The results wereoutlined as follows:1.The alveolar casts were observed from the subpleural and intralobular septu-lar surfaces.The human alveolus is irregular polygon in appearance.The size ofthe alveolus is variable,its surface is smooth,there are many depressions of the topof type Ⅱ cells.The bridge-like structure between two adjacent alveoli are thecasts of the alveolar pores.They are variable both in size and number,and ofround or oval shapes with smooth surface.2.Capillaries of the subpleural space and interlobular septulum are transitional,and identical in appearance.Meshes of the capillary network are larger than thoseof other parts,but more closer in density compared with those in dog.Each capil-lary is branched from the metaarteriole.3.Capillaries in the alveolar septum is a single layer of dense network,theirdiameter are larger than those of the mesh hole.They originate mainly from thecapillaries of subpleural space and interlobular septulum.

Objective:To investigate the clinicopathological and molecular characteristics of pulmonary enteric adenocarcinoma (PEAC).Methods:The clinical and pathological data of 19 cases of PEAC in the Affiliated Cancer Hospital of Zhengzhou University were retrospectively collected from 2015 to 2019. Immunohistochemistry (IHC) was used to detect the relevant immunophenotypes, amplification refractory mutation system (ARMS) and fluorescence in situ hybridization (FISH) were used to detect the expression of EGFR, KRAS and ALK genes. The patients were followed up, and the relevant literature was reviewed and analyzed.Results:There were 19 cases, including 10 males and 9 females, with a mean age of 58 years (range 33-71 years). Microscopically, the tumors showed moderately to highly differentiated adenoid and/or papillary growth patterns. The tumor cells were highly columnar and sometimes showed pseudostratification. Inflammatory necrosis and scattered nuclear fragmentation were seen in some glandular lumens. IHC showed variable expression of CK7 (19/19), TTF1 (8/19), Napsin A (6/19), villin (17/19), CK20 (16/19) and CDX2 (10/19). Molecular testing showed KRAS mutation in nine cases (9/19), EGFR mutation in one case (1/19), and positive ALK split signal in one case (1/19). In the literature, the reported mutation rate of KRAS in PEAC was much higher than that of EGFR and ALK. All 19 cases underwent surgical resection and 11 cases were subjected to chemotherapy or radiotherapy.Conclusions:PEAC is a rare variant of invasive pulmonary adenocarcinoma, and has similar histological and cytological features to that of colorectal adenocarcinoma. However, detailed medical history, histologic heterogeneity, an IHC combination of CK7 +/villin + and high KRAS mutation rate are the key points of diagnosis. The prognosis needs long-term follow-up and big data statistics.

Objective:To observe the curative effects of berberine in rats with high-fat diet induced non-alcoholic fatty liver and to further explore its possible mechanism.Methods:Twenty-six Sprague-Dawley rats (120-160 g) were randomly divided into 3 groups: control group ( n = 8), model group ( n = 10) and treatment group ( n = 8). Rats in the control group were fed with regular diet, and the model group and the treatment group were fed a high-fat diet. At the 12th week, two rats in the in the model group were sacrificed to verify whether model was successful established. Subsequently, treatment group rats were given a gavage of berberine at a dose of 150 mg·kg -1·d -1 for 4 weeks, and the control and the model group rats were given the same dose of normal saline. Rats were sacrificed at week 16th. HE staining was used to observe the changes in the intestinal mucosa of rats. Sudan black B staining was used to observe the fatty changes in liver. Immunohistochemical staining was used to observe the expression level of occludin protein in the intestinal epithelium. A real-time 16S rDNA PCR method was used to measure the number of escherichia coli, bacteroides and faecalibacterium prausnitzii in the feces of rats. Results:Model group had a higher serum levels of endotoxin (0.288 ± 0.045) and tumor necrosis factor (TNF)-α (1.07 ± 0.11) than the control group (0.192 ± 0.049, 0.94 ± 0.07) ( P < 0.05). Berberine intervention had significantly reduced endotoxin (0.213 ± 0.025) and TNF-α level (0.93 ± 0.07) ( P < 0.05). The expression level of occludin protein was significantly lower in the intestinal mucosa of model group than that of control group (0.166 ± 0.014), and berberine had promoted the expression of occludin protein in intestinal mucosa (0.055 ± 0.009), but the difference was not statistically significant ( P > 0.05). At the same time, compared with the model group (7.29 ± 0.47), the number of bacteroidetes in the control group (9.49 ± 0.59) was decreased, while the number of bacteroidetes in the treatment group was increased (9.77 ± 0.87). The number of escherichia coli (6.92 ± 0.77) and faecalibacterium prausnitzii (8.70 ± 0.62) in the model group were increased than control group (5.42 ± 0.63, 9.49 ± 0.59), while the number of escherichia coli (6.34 ± 0.71) and faecalibacterium prausnitzii (9.77 ± 0.87) ( P < 0.05) was reduced with the intervention of berberine. Conclusion:Berberine could effectively protect the intestinal barrier function in rats with NAFLD and the possible mechanism of action behind it may be the regulation of intestinal flora.

Objective: To investigate the effect of curcumin on intestinal mucosal mechanical barrier in rats with non-alcoholic fatty liver disease. Methods: A total of 30 male Sprague-Dawley rats were equally divided into normal control group, model group, and curcumin intervention group. The rats in the model group and the curcumin intervention group were given high-fat feed for 16 weeks, and those in the curcumin intervention group were given curcumin 200 mg/kg/day by gavage once a day after 8 weeks of high-fat feeding. The rats were sacrificed at the end of week 16. A light microscope was used to observe pathological changes in the liver, an electron microscope was used to observe the tight junction of the intestinal mucosa, an automatic biochemical analyzer was used to measure the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), chromogenic substrate Limulus amebocyte lysate assay was used to measure plasma lipopolysaccharide (LPS) level, spectrophotometric method was used to measure the activity of serum diamine oxidase, ELISA was used to measure the serum level of tumor necrosis factor-α (TNFα), and immunohistochemistry was used to measure the expression of the tight junction protein occludin. One-way ANOVA test and SNK-q test were used for statistical analysis. Results: Under the light microscope, the control group had no hepatocyte steatosis, the model group had significant hepatocyte steatosis and inflammatory cell infiltration, and the curcumin intervention group had reduced hepatocyte steatosis and inflammatory cell infiltration. Under the electron microscope, the control group had a clear and complete structure of the tight junction of the intestinal mucosa and normal structures of mitochondria and endoplasmic reticulum; in the model group, the structure of the tight junction of the intestinal mucosa was destroyed, the intercellular space was widened, the desmosomes had a loose structure, there was edema in some mitochondria, and the endoplasmic reticulum was dilated; the curcumin intervention group had improvements in the structure of tight junction of the intestinal mucosa, intercellular space, edema in the mitochondria, and dilation of the endoplasmic reticulum. Compared with the control group, the model group had significant increases in the serum levels of AST, ALT, DAO, TNFα, and LPS (q = -15.918, -14.402, -33.700, -8.944, and -10.832, P < 0.05); compared with the model group, the curcumin intervention group had significant reductions in the serum levels of AST, ALT, DAO, TNFα, and LPS (q = 10.457, 7.752, 18.802, 5.202, and 4.279, P < 0.05). In the control group, occludin showed a linear distribution along the top of small intestinal mucosal epithelial cells. The model group had a significant reduction in positive staining compared with the control group, and the curcumin intervention group had a significant increase in positive staining compared with the model group. The relative expression of occludin was 0.29±0.03 in the control group, 0.12±0.02 in the model group, and 0.21±0.02 in the curcumin intervention group (P < 0.05). Conclusion Intestinal mucosal mechanical barrier is impaired in rats with NAFLD. Curcumin can reduce such damage, and its mechanism of action may be related to up-regulating the expression of occludin in the intestinal mucosa and reducing the levels of TNFα and LPS.