Objective To observe the expression level of macrophage stimulating protein (MSP) in acute on-chronic liver.failure (ACLF) patients,and to explore the clinical significance and correlation with different immune regulatory factors.Methods The double antigen sandwich enzyme-linked immunosorbent assay method was used to detect MSP in the peripheral blood of 45 patients who were diagnosed with ACLF and 32 cases of chronic hepatitis B (CHB).The MSP levels were compared among ACLF patients with different outcomes,and the MSP level of healthy person was used as control.Meanwhile,liver function and hepatitis B virus (HBV) load were detected,and the expressions of peripheral blood CD4+ interferon (IFN)γ+ (helper T cell 1 Th1),CD4+ interleukin (IL)-4+ (helper T cell 2,Th2),CD4+IL-17+ (helper T cell 17,Th17) and CD4+ CD25+ Foxp3+ (regular T cell,Treg) were measured by flow cytometry.The comparison of means between two samples was done by t test,and oneway ANOVA and linear correlation analysis were also used.Results The serum MSP levels in ACLF patients,CHB group and healthy control were (1.65±0.46) ng/mL,(1.43±0.32) ng/mL and (1.23±0.21) ng/mL,respectively.The serum MSP level in ACLF patients was significantly higher than both CHB patients (t=2.163,P=0.035) and healthy control (t=4.032,P=0.01).The MSP level in ACLF survival group was statistically higher compared with ACLF death group ([2.29 ± 0.42] ng/mL vs [1.42±0.17] ng/mL,t=1.973,P=0.042).Th2,Th17 cells in ACLF group,CHB group and healthy control group were (1.51±0.27) ％ and (1.94±1.02)％,(0.42±0.08)％ and (0.55±0.36)％,(0.23±0.19) ％ and (0.26±0.19) ％,respectively,which were all significantly different (F=7.759 and 37.229,respectively;both P＜0.01).The MSP level was positively associated with the number of Th2 (r=0.386,P=0.032) and Th17 (r=0.644,P=0.000),and the ratio of Th17/Treg (r =0.605,P=0.000);while it was negatively associated with the number of Th1 (r=-0.212),Treg (r=-0.262) and the ratio of Th1/Th2 (r=-0.394) (all P＞0.05).Conclusion MSP is involved in the progress of ACLF,and it may be associated with clinical outcomes and cellular immune imbalance of ACLF patients.

Intrahepatic cholestasis of pregnancy (ICP) is the most common liver disease during pregnancy. ICP usually occurs in late pregnancy, with main manifestations of pruritus, jaundice, and increases in serum levels of total bile acid and aminotransferases found by laboratory examination. ICP has great impact on the fetus and may lead to preterm birth, stillbirth, and deformity. ICP has complex etiologies, and studies have shown that genetic, metabolic, immunological, and hormonal factors may contribute to the onset of ICP. The genes associated with cholestasis, such as ABCB11, ABCB4, ATP8B1, and FXR, are associated with the genetic susceptibility of ICP. This article reviews the association between cholestasis-related genes and ICP.

Objective: To identify the type and origin of ATP7B gene mutation in a family affected with Wilson disease by combined use of multiple methods. Methods: Peripheral blood samples were collected from the proband, her parents and her brother. Sanger sequencing were used to detect point mutation and small deletion/insertion of the 21 exons and flanking sequences of the ATP7B gene in all family members. Array-based comparative genomic hybridization (aCGH) was performed to identify copy number variations (CNVs) of the ATP7B gene in the proband. The result was validated by quantitative PCR (qPCR) in other 3 members. Results: Sanger sequencing indicated that the proband carried a heterozygous variation c. 2668G>A (p.V890M) derived from her mother. In addition, 5 common SNPs were detected in her mother, three of which were also identified in her father and brother. The 5 SNPs in the proband were of the wide type. aCGH analysis demonstrated that the proband was heterozygous for a 4 kb deletion, which encompassed exons 2 and 3 of the ATP7B gene and 2 SNPs. qPCR showed that the copy number in her father and brother was about half of the control, indicating heterozygous loss of exons 2 and 3. Conclusion The combined Sanger sequencing, array CGH and qPCR has identified a novel CNV involving the ATP7B gene. The strategy can improve the diagnostic rate for hereditary or rare diseases.