Association of Hepatitis A exposure and TIM-1 with childhood allergic asthma.

BACKGROUND: Hepatitis A virus (HAV) receptor (TIM-1) polymorphism plays an important role in asthma and autoimmune diseases. Objective. To analyze the association of TIM-1 polymorphism and HAV infection with childhood allergic asthma in Southwest China. METHODS: TIM-1 exon 4 (157insMTTTVP) and two polymorphism loci, -416G>C and -1454G>A, in the HAV receptor promoter region were studied. Polymerase chain reaction (PCR) was used to test the genotypes of three polymorphism loci among 579 cases of asthma and 524 controls. The HAV infection status was determined in a case-control study with stratified analysis. RESULTS: HAV exposure associated with childhood allergic asthma in the study population was compared with controls (odds ratio (OR) = 0.181, 95% confidence interval (CI) 0.126-0.260, p < .001). The -416G>C polymorphism was associated with asthma (OR = 1.384, 95% CI 1.148-1.669, p < .001), but the insertion variant 157delMTTTVP of exon 4 and the -1454G>A polymorphism were not. CONCLUSION: Our results indicated that the -416G>C polymorphism of the TIM-1 gene is associated with childhood allergic asthma, providing a better understanding of the pathogenesis of the allergic asthma among children aged below 15 years in Southwest China.

[Relationship between TIM-4 polymorphism and childhood asthma].

OBJECTIVE: To elucidate whether the polymorphism of asthma immune regulator gene TIM-4 is associated with the risk of childhood allergic asthma in the southwest region of China. METHODS: TIM-4 gene promoter region RS6882076 and intron RS4704727 were studied. PCR-RFLP was used to test the genotypes of two polymorphism loci among 579 cases (average 7.2 years old) of asthma and 524 controls (average 7.6 years old) in a case-control study. RESULTS: There were significant differences in the frequency of gene types at RS4704727 site between the asthma and the control groups (P<0.01). The results of PCR-RFLP showed that the polyporphisms of RS6882076 and RS4704727 in TIM-4 gene were present in this study population. The frequency of T allele at the RS4704727 site in the asthma group was significantly lower than that in the control group (OR=1.603; 95%CI 1.304-1.971; P<0.01). There were no significant differences in the frequencies of gene types and allele at RS6882076 site between the two groups (P>0.05). CONCLUSIONS: RS4704727 polymorphism of TIM-4 gene may be associated with childhood asthma, providing a better understanding of the pathogenesis of childhood asthma in the Southwest region of China.

[Human fetal liver nonparenchymal mesenchymal stem cells differentiate into functional hepatocyte-like cells in vitro].

OBJECTIVE: To induce nonparenchymal mesenchymal stem cells (NPMSCs) differentiating into functional hepatocyte-like cells in vitro, and to identify the molecular biology and functional characteristics of those hepatocyte-like cells. METHODS: Human NPMSCs were isolated and cultured with cell culture technique. NPMSCs were induced (on 1% Matrigel as a matrix and then submitted to 2.5 mmol/L AZA pretreatment for 10-12 h), by adding HGF 10 microg/L + FGF4 10microg/L + HGM into the culture medium. The characteristics of proliferation and growth of human NPMSCs were studied with methyl thiazolyl tetrazolium (MTT). The phenotypes of NPMSCs were identified by flow cytometry, immunohistochemistry and RT-PCR. Albumin (Alb) levels in culture supernatants were determined with ELISA. Staining for glycogen of undifferentiated NPMSCs and NPMSCs derivated hepatocyte-like cells was conducted with periodic acid-Schiff (PAS) test. RESULTS: Growth and division of adherent cells obtained from fetal livers were good and the amount of NPMSCs resourced from each fetus could be amplified to 109 cells after 10 serial subcultivations. The phenotype of NPMSCs was CD166 positive and CD34 negative. The shape of NPMSCs plated on Matrigel with FGF4 and HGF changed from long fusiform to polygonal or round on days 21-28. The rate of cell rounding was 40% and the ratio of dikaryocytes was 5%. Immunohistochemical and RT-PCR detection showed that undifferentiated NPMSCs expressed few alpha fetoprotein (AFP) and AFP mRNA, and did not express any of the liver-specific transcription factors or cytoplasmic markers. Many cells in early induction expressed GATA4, AFP and CK18 proteins and their mRNAs, and their expressions were reduced at the late induction, but the expressions of Alb, CK18, GST-and hepatocyte transcription factor HNF1increased gradually. The ratio of Alb and CK18 positive cells was 60%. Undifferentiated NPMSCs did not produce Alb. Alb production by induced NPMSCs increased in a time-dependent manner. Glycogen storage was first seen on day 14, and maximum levels were seen after day 28. CONCLUSIONS: There are MSCs among nonparenchymal cells of fetal livers. A high ratio of hepatocyte-like cells was obtained under our induction condition. NPMSCs differentiate firstly into hepatocyte precursors, and then differentiate into mature hepatocytes and hepatocyte-like cells with positive hepatocyte markers. The induced NPMSCs have hepatocyte specific functional features.

[Isolation, cultivation and biological identification of human fetal marrow mesenchymal stem cells].

OBJECTIVE: Noting the morphological and cytobiology characteristics and phenotypes of MMSCs, to establish an isolation and culture method for fetal MMSCs in order to provide a source of marrow mesenchymal stem cells (MMSCs). METHODS: Fetal MMSCs were isolated and cultured with in vitro cell culture technique; the characteristics of the proliferating and growing fetal MMSCs were studied with MTT and image analysis; the phenotypes of MMSCs were identified by flow cytometry and immunohistochemistry. RESULTS: Bone marrow of 12 fetuses was isolated within 0.5-2 h, and about 300+/-80 adherent cells were obtained at 24 h. Colonies with more than 5 cells were 15+/-6, growth detention period of culture cell was at 1-3 d after planting, log phase growth period was at day 4, and the amount of disintegration phase cells was reduced significantly. Original culture and serial subcultivations showed that cells divided prosperiously; unequal divisions special for stem cells were observed, and the amount of MMSCs harvested from each fetus was as much as 10(11)-10(12) cells after 10 serial subcultivations. The phenotype of MMSCs was CD166 positive and CD34 negative. Serial subcultivated MMSCs expressed a microamount of AFP and did not express albumine or CK18. CONCLUSION: Fetal MMSCs are easily isolated and proliferate prosperouly. Serial subcultivated MMSCs did not differentiate into hepatocyte-like cells under common culture condition and are feasibile as seed cells for tissue engineering reconstruction.

Effects of hemoperfusion adsorption and/or plasma exchange in treatment of severe viral hepatitis: a comparative study.

AIM: Non-bioartificial liver has been applied to clinic for quite a long time, but the reported efficacy has been very different. The aim of this study was to compare the efficacy and safety of hemoperfusion adsorption, plasma exchange and plasma exchange plus hemoperfusion adsorption in treatment of severe viral hepatitis. METHODS: Seventy-five patients with severe viral hepatitis were treated with hemoperfusion adsorption therapy (24 cases), plasma exchange therapy (17 cases) and plasma exchange plus hemoperfusion adsorption therapy (34 cases). The data of liver function, renal function, blood routine test, prothrombin time (PT) and prothrombin activity (PTa) pre- and post-therapy were analyzed. RESULTS: Clinical symptoms of patients improved after treatment. The levels of aminotransferase, total bilirubin, direct bilirubin decreased significantly after 3 therapies (P<0.05 or P<0.01). PT, the level of total serum protein decreased significantly and PTa increased significantly after plasma exchange therapy and plasma exchange plus hemoperfusion adsorption therapy (P<0.05 or P<0.01). The side effects were few and mild in all patients. CONCLUSION: Three therapies were effective in the treatment of severe viral hepatitis. Plasma exchange therapy and plasma exchange plus hemoperfusion adsorption therapy are better than hemoperfusion adsorption therapy.

[Preliminary evaluation on the effects of a hybrid bioartificial liver support system in the treatment of hepatic failure].

OBJECTIVES: To construct a novel hybrid artificial liver support system and evaluate its clinical efficacy in the treatment of hepatic failure. METHODS: A hybrid bioartificial liver support system consisting of plasma exchange device, charcoal perfusion column, and bioreactor cultured human or porcine hepatocytes was developed. 30 patients with hepatic failure were treated using this hybrid system. RESULTS: Both the excellent rate and effectual rate of the artificial liver support system were 43.3% (13/30). The total effectual rate was 86.7%. Finally, eleven out of 30 patients recovered completely. Six patients were bridged to liver transplantation. Six patients (20%) died of hepatic failure and seven patients (23.3%) discharged due to worsening of disease. CONCLUSIONS: The hybrid artificial liver support system has prominent liver support effects for hepatic failure, which can be regarded as an efficient measure for the treatment of severe hepatitis.

[Effects evaluation of middle mode artificial liver in treatment of severe hepatitis].

OBJECTIVE: To evaluate the clinical efficacy and safety of middle mode artificial liver-plasma pheresis in treatment of severe hepatitis. METHODS: Seventeen patients with severe hepatitis were treated with plasma pheresis. The results of liver function, renal function, blood routine, prothrombin time (PT), prothrombin time activity (PTa) before and after the treatment were analyzed. All patients were observed closely. RESULTS: Symptoms of patients treated with plasma pheresis were improved, and the total effective rate reached 58.8 percent, but the survival rate was only 11.8 percent. Compared with those before the therapy, there were significant differences in aminotransferase, total bilirubin, direct bilirubin, PT, PTa and total protein level after treatment (P<0.05 or P<0.01). The side-effects were mild. CONCLUSION: Middle artificial liver is effective in the treatment of severe hepatitis.