Objective To identify the key immune-related genes and potential drug targets of Krabbe disease by using neural stem cells(NSCs)derived from induced pluripotent stem cells(iPSCs)generated from patients with Krabbe disease.Methods 1)The transcriptome data of GLD-NSCs(GSE212512)was analyzed using GEO2R and Sangerbox;2)Differential expression genes(DEGs)related to immunity were identified through various methods;3)Functional enrichment and pathway analysis were performed using DAVID;4)The protein-protein interaction network was constructed and analyzed using STRING and Cytoscape;5)Key immune-related genes were deter-mined through topological analysis;6)Potential therapeutic drugs were screened using DGIdb and drug MAP data-bases;7)Molecular docking was performed using CB-DOCK2.Results 1)Sixty-one immune-related DEGs were identified in patient NSCs,with GDNF,EGF,KDR,FGF10,and MET being identified as key immune-related genes;2)Drug candidates with high binding affinity to these genes were identified,including gentamicin for GDNF,cetuximab for EGF,tivozanib for KDR,and capmatinib for MET.Conclusions The predictions suggest cetuximab,gentamicin,tivozanib,and capmatinib exhibit high binding affinity with EGF,GDNF,KDR,and MET,respectively,indicating their potential application value in GLD targeted therapy.

Objective To investigate the effects and mechanism of Dahuang Lingxian Prescription on bile duct reaction of cholestatic liver fibrosis rats caused by bile duct ligation.Methods A total of 40 SD rats were randomly divided into blank group,model group,ursodeoxycholic acid group and Dahuang Lingxian Prescription group,with 10 rats in each group.Except for the blank group,the remaining groups of rats underwent bile duct ligation surgery to establish a cholestatic liver fibrosis model.After surgery,the ursodeoxycholic acid group was given ursodeoxycholic acid solution by gavage,Dahuang Lingxian Prescription group was given Dahuang Lingxian Prescription solution by gavage,and the blank group and model group were given equal volume of normal saline by gavage,once a day for 3 consecutive weeks.The activities of serum AST,ALT,ALP,GGT and the contents of TBIL,TBA were tetected,the morphology of liver tissue was observed by HE staining,and the liver fibrosis was observed by Masson staining,immunohistochromic staining and Western blot were used to detect the expressions of CK19,CK7,EpCAM and SOX9 proteins.Results Compared with the blank group,the liver surface of the model group rats was rough,with a harder texture and obvious graininess,HE staining showed damage to the liver lobule structure,forming pseudo lobules,a large number of bile duct hyperplasia and inflammatory cell infiltration,and a significant increase in collagen fiber deposition(P<0.01);the activities of serum AST,ALT,ALP,GGT,as well as the contents of TBIL and TBA significantly increased(P<0.01);the positive expressions of CK19,CK7 and EpCAM in liver tissue significantly increased(P<0.01),and the protein expressions of CK19,CK7,EpCAM and SOX9 significantly increased(P<0.01).Compared with the model group,the appearance and texture of the liver of the rats in the ursodeoxycholic acid group and Dahuang Lingxian Prescription group were relatively softer,the lobular structure was less damaged,the inflammatory cells infiltration was less,the collagen fiber deposition was significantly reduced(P<0.01),the activities of serum AST,ALT,ALP,GGT,and the contents of TBIL and TBA were significantly decreased(P<0.01);the expressions of TBA and TBIL were significantly decreased(P<0.01),the positive expressions of CK19,CK7 and EpCAM significantly decreased(P<0.01),and the protein expression of CK19,CK7,EpCAM and SOX9 significantly decreased(P<0.01).Conclusion Dahuang Lingxian Prescription can inhibit the bile duct reaction associated with liver progenitor cells,decrease the expression of CK19,CK7,EpCAM and SOX9,and thus improve the cholestatic liver fibrosis of rats induced by bile duct ligation.

Objective To investigate the effects and mechanism of Dahuang Lingxian Prescription on bile duct reaction of cholestatic liver fibrosis rats caused by bile duct ligation.Methods A total of 40 SD rats were randomly divided into blank group,model group,ursodeoxycholic acid group and Dahuang Lingxian Prescription group,with 10 rats in each group.Except for the blank group,the remaining groups of rats underwent bile duct ligation surgery to establish a cholestatic liver fibrosis model.After surgery,the ursodeoxycholic acid group was given ursodeoxycholic acid solution by gavage,Dahuang Lingxian Prescription group was given Dahuang Lingxian Prescription solution by gavage,and the blank group and model group were given equal volume of normal saline by gavage,once a day for 3 consecutive weeks.The activities of serum AST,ALT,ALP,GGT and the contents of TBIL,TBA were tetected,the morphology of liver tissue was observed by HE staining,and the liver fibrosis was observed by Masson staining,immunohistochromic staining and Western blot were used to detect the expressions of CK19,CK7,EpCAM and SOX9 proteins.Results Compared with the blank group,the liver surface of the model group rats was rough,with a harder texture and obvious graininess,HE staining showed damage to the liver lobule structure,forming pseudo lobules,a large number of bile duct hyperplasia and inflammatory cell infiltration,and a significant increase in collagen fiber deposition(P<0.01);the activities of serum AST,ALT,ALP,GGT,as well as the contents of TBIL and TBA significantly increased(P<0.01);the positive expressions of CK19,CK7 and EpCAM in liver tissue significantly increased(P<0.01),and the protein expressions of CK19,CK7,EpCAM and SOX9 significantly increased(P<0.01).Compared with the model group,the appearance and texture of the liver of the rats in the ursodeoxycholic acid group and Dahuang Lingxian Prescription group were relatively softer,the lobular structure was less damaged,the inflammatory cells infiltration was less,the collagen fiber deposition was significantly reduced(P<0.01),the activities of serum AST,ALT,ALP,GGT,and the contents of TBIL and TBA were significantly decreased(P<0.01);the expressions of TBA and TBIL were significantly decreased(P<0.01),the positive expressions of CK19,CK7 and EpCAM significantly decreased(P<0.01),and the protein expression of CK19,CK7,EpCAM and SOX9 significantly decreased(P<0.01).Conclusion Dahuang Lingxian Prescription can inhibit the bile duct reaction associated with liver progenitor cells,decrease the expression of CK19,CK7,EpCAM and SOX9,and thus improve the cholestatic liver fibrosis of rats induced by bile duct ligation.

Objective:To determine the seroprevalence of celiac disease in susceptible population, and to analyze the relationship between demographic characteristics, dietary habits, lifestyle and serological positivity so as to provide guidance for the prevention and treatment of celiac disease in Southern China.Methods:A total of 1 273 individuals who participated in Guangdong Province Health Screening Program in 2015, were selected as serologically positive subjects of celiac disease, including people with irritable bowel syndrome, colitis, diarrhea, anemia, low BMI, short stature, type 1 diabetes mellitus (T1DM), rheumatoid arthritis (RA), ankylosing spondylitis, psoriasis and bristol grade=6 or 7. All subjects were tested for serum IgA anti-tissue transglutaminase antibodies (TTGA), IgA antibodies against deamidated gliadin peptides(DGPA) and IgG against deamidated gliadin peptides (DGPG). Dietary habits, lifestyle and demographic characteristics were compared in subgroups.Results:The seroprevalence of celiac disease in susceptible population was 0.94% (95% CI 0.54%-1.64%) including 0.08% (1/1 273) for TTGA, 0.47% (6/1 273) for DGPA, and 0.39% (5/1 273) for DGPG. The seropositive rate was 3.6% (1/28) in patients with psoriasis, 2.1% (2/95) in the low BMI group, 1.9% (1/53) in T1DM group, 1.8% (3/169) in diarrhea group and 1.1% (5/463) in RA group. No significant difference was found in age, gender, high carbohydrate diet or lifestyle between the negative and the positive subjects. Conclusions:In Southern China, the seropositive rate of celiac disease is 0.94% in susceptible population, which prompts an urgent need of serological screening for early diagnosis.

Objective:To study microwave-assisted extraction technique of flavonoids from Ophiopogon japonicus. Methods:The effect of the parameters,i.e.ratio of solid to liquid,microwave treatment time,microwave power and volume ratio of ethanol on extraction amount of flavonoid were analyzed by single factor experiment.Results:Optimum processing data were determined as follows:90.0% ethanol as extracting solvent,stock ratio 1:14(g/ml) ,microwave extracting for 4 minutes at the power of MED. The extraction rate was 39.7% higher than that of non-microwave's.Conclusion:Extraction of flavonoids by microwave from Ophiopogon japonicus is an eficient method.

Objective To observe effects of four extracts (water, n-butanol, acetoacetate and petro-leum benzine extracts) from Hippocampus japonicus on kidney-yang deficiency mice induced by extradiol, and reveal the modern scientific connotation about sweet-flavor, warm-nature, meridian tropism in liver and kidney. Methods Kidney-yang deficiency mice was induced by intraperitoneal injection of estradiol benzoate. Effects of four extracts from H. japonicus were observed by measuring physical signs, rectal temperature, grip strength, swimming time, autonomic mobility, creatinine (Cr) and urea (UR) level in serum, haematological indexes, testis index and seminal vesicle index. Results All the four extracts from H. japonicus could markedly improve physical signs and increase seminal vesicle index, RBC, Hb and PLT in blood, and decrease UR level in serum of kidney-yang deficiency mice, water extract and petoleum benzine extract could increase rectal temperature, grip strength, swimming time, autonomic mobility, testis index, WBC, lymphocyte (LY), monocyte (MO), and neutrophilic granulocyte (NE), n-butanol extract could increase autonomic mobility and NE, acetoacetate extract could enhance grip strength and prolong swimming time. Conclusion All above effects of extracts from H. japonicus could be as the basic of improving kidney-yang deficiency, could be an important part of the modern scientific connotation about sweet-flavor, warm-nature, meridian tropism in liver and kidney. Effects of water extract and petroleum ether extract were closer and stronger, which offer enlightenment for searching the common effective substance groups of H. japoniacs.

Objective: To observe antipyretic, diuresis, anti inflammatory and antisepsis effects of Qinglin Granules. (Herba Polygoni Avicularis, Herba Dianthi, Caulis Aristolochiae Manshuriensis, Semen Plantaginis, Fructus Gardeniae, etc.) Methods: The model of pyretic rats induced by yeast was used to prove antipyretic action. The diuresis action was observed by the test. The anti inflammatory effect was observed by pettitoes swelling in rats and auricle swelling in mice. The paper diffuse methods were used to confirm the antisepsis effect. Results: Qinglin Granules not only had the obvious effect of relieving fever and increasing the amount of urine, but also had the strong inhibition on the two kinds of inflammatory models, moreover it can widen the diameter of the antisepsis ring which showed it had the inhibition on cdibacillus, pseudomonas, aeruginosa, bacillus gasoformans, staphylococcus aureus and gonococcic. Conclusion: Qinglin Granules have the obvious antipyretic, diuresis, anti inflammatory and antisepsis effects.