1.Study on in vitro model of hepatitis B virus-infected transwell chambers mediated by peripheral blood mononuclear cells
Junni WEI ; Yue ZHANG ; Xuefeng GAO ; Shulian XUE ; Suping WANG
Chinese Journal of Infectious Diseases 2015;(6):347-350
Objective To observe the transport of hepatitis B virus (HBV)-infected peripheral blood mononuclear cells (PBMC)through placental barrier set up by choriocarcinoma trophoblast cells (Bewo cells),and to explore the biological role of PBMC as a carrier for HBV transport.Methods Bewo cells and PBMC were cultured and their proliferation and activity were detected by cell counting kit (CCK)-8.One hundred μL serum containing 5 ×10 6 copy/mL HBV DNA was used to infect PBMC,and cells infected with HBV were labeled by fluorescent dye carboxyfluorescein diacetate succinimidyl ester (CFSE).A co-culture model of Bewo cells and HBV-infected PBMC was set up by transwell chamber. The migration of HBV-infected PBMC was detected by flow cytometry.Realtime fluorescence quantitative polymerase chain reaction method was used to detect HBV DNA contents of PBMC under transwell chamber.Results PBMC and Bewo cells proliferated at around 24 h and entered into growth stagnation at around 120 h.The contents of PBMC labeled by green fluorescent at 0,12,24 and 48 h during co-culture under chamber were (0.445 ±0.021)%,(21 .180 ± 4.653 )%,(34.830 ± 7.156 )% and (64.185 ± 3.161)%,respectively.The amount of PBMC marked green fluorescence increased over prolonged incubation time (F =68.983,P =0.001 ).PBMC HBV DNA contents at 24 and 48 h of co-culture under chamber were (1.925±0.431)×103 copy/mL and (2.565 ±0.361)×103 copy/mL,respectively,indicating that PBMC under chamber were infected with HBV.Conclusions PBMC may be a target for HBV infection in extrahepatic tissues.Placental trophoblastic barrier built by transwell chambers may provide new ideas to investigate HBV transmission across the placenta in vitro .
2.Relationship between placenta HBsAg and hepatitis B serum markers in newborns
Junni WEI ; Yue ZHANG ; Shulian XUE ; Junfeng ZHANG ; Jieyu SHUANG ; Liping FENG ; Suping WANG
Chinese Journal of Infectious Diseases 2012;(11):668-672
Objective To study the relationship between placenta HBsAg in HBsAg positive pregnant women and serum hepatitis B virus (HBV) markers,HBV DNA levels in newborns.Methods Placenta HBsAg was detected by immunohistochemical affinity hormone-biotin complex (ABC) method in 155 HBsAg positive pregnant women.Serum HBV markers in newborns were detected by enzyme-linked immunosorbent assay (ELISA).Serum HBV DNA levels of newborns were detected by real-time fluorescence quantitative polymerase chain reaction (PCR).The positive rates were compared using x2 test.Results HBsAg was expressed with different levels in various types of cells of placenta in 155 pregnant women.The total placenta HBsAg positive rate was 37.4% (58/155),and those in decidual cells,trophoblastic cells,villous mesenchymal cells and villous capillary endothelial cells were 37.4% (58/155),25.8% (40/155),18.7% (29/155) and 7.1% (11/155),respectively.The HBsAg positive rates of placenta gradually decreased from decidual cells of the maternal surface to villous capillary endothelial cells of the fetal surface (tendency x2 =43.01,P=0.00).The positivity of placenta HBsAg was associated with both HBsAg and HBeAg in newborns (x2 =4.88,P<0.05 and x2 =3.86,P<0.05,respectively),while that was not associated withanti-HBe and anti-HBc in newborns (x2 =3.36,P>0.05 and x2 =0.00,P> 0.05,respectively).The risk of HBsAg positive in newborns was higher when HBsAg was positive in villous capillary endothelial cells and villous mesenchymal cells (OR=5.31,95 %CI=1.38-20.40 and OR=3.33,95%CI=1.16-9.52,respectively).The risk of HBeAg positive in newborns was higher when HBsAg was positive in trophoblastic cells and villous mesenchymal cells (OR=3.04,95 %CI=1.45-6.39 and OR=3.05,95 % CI=1.32-7.03,respectively).However,placenta HBsAg positive was not associated with HBV DNA positive in newborns (x2 =0.09,P>0.05).Conclusion The risk of neonatal HBV serological markers positive is higher when the HBsAg positive placental cells are closer to fetal surface,which indicates that HBsAg enters fetal blood circulation by means of cell transferring layer by layer.
3.Changes of metallothionein contents in plasma and tissues of rabbit with atherosclerosis
Shulian LI ; Xiaohong WANG ; Lin XUE ; Jing LIN ; Chunyu ZHAO ; Chaoshu TANG
Chinese Journal of Pathophysiology 2000;0(07):-
AIM: To observe the level of metallothionein (MT) in liver, aorta and plasma of rabbit with atherosclerosis (AS) in order to recognize the alteration of oxidative defense system in body when AS occurred.METHODS:Preparation of AS model of rabbit induced by having high-fat diet for eight weeks; the levels of MT and malondialdehyde (MDA) were measured in the tissues of liver and aorta and plasma of rabbit.RESULTS:The MT levels in liver tissues and plasma in atherosclerotic group increased 318%( P
4.Homocysteine induces metallothionein formation in mice
Shulian LI ; Zaiquan LI ; Ying ZHANG ; Lin XUE ; Song XU ; Xiaohong WANG ; Chaosh TANG
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To observe the changes of metallothionein (MT) in various tissues of mice during hyperhomocysteinemia. METHODS: Intraperitoneal injection of homocysteine into mice induced hyperhomocysteinemia. The contents of tissue MT and malondialdehyde (MDA) in liver, heart and kidney were determined. RESULTS: Compared with control group, tissue MT levels in Hcy-group animals were increased by 210% ( P 0.05),respectively, compared with Hcy alone group. Tissue MDA contents were decreased by 24% ( P
5.Clinical outcome and prognostic factors of primary gastric mucosa-associated lymphoid tissue lymphoma: a retrospective analysis of 77 cases
Shulian WANG ; Liyan XUE ; Yongwen SONG ; Jing JIN ; Weihu WANG ; Yueping LIU ; Xinfan LIU ; Zihao YU ; Ning Lü ; Yexiong LI
Chinese Journal of Radiation Oncology 2009;18(2):105-109
Objective To analyze the clinical results and prognostic factors of patients with early-stage primary gastric mucosa-associated lymphoid tissue(MALT) lymphoma. Methods Seventy-seven pa-tients with primary gastric MALT lymphoma treated from 1985 to 2006 were retrospectively analyzed. All pa-tients were pathologically confirmed as MALT lymphoma in stage Ⅰ ,Ⅱ and ⅡE (by modified Blackedge staging system). Thirty-seven patients had stage Ⅰ disease,23 stage Ⅱ and 17 stage ⅡE. Sixty patients un-derwent surgical resection and 17 received non-surgical treatment. Survival rates were calculated by the Kap-lan-Meier analysis with the Logrank test. Results With a median follow up of 57 months for the surviving patients(ranging from 1 to 198 months for all patients), the 5-year overall survival rate, disease-free survival rate,loco-regional control rate and distant metastasis free survival rate were 74% ,70% ,76% and 87% ,re-spectively. In univariate analysis, clinical stage was significantly associated with overall survival. Patients with stage Ⅰ or Ⅱ disease had a better overall survival than those with stage ⅡE (P = 0.01). Tumor size and surgical resection were significantly associated with disease-free survival. Patients with primary tumor 8 cm or less in diameter had better disease-free survival than those with primary tumor more than 8 cm in diameter(P =0.03). Patients who underwent complete resection had better disease-free survival than those who under-went incomplete resection or no surgery (P =0.02). Clinical stage, tumor size and surgical resection were significantly associated with loco-regional control. Patients with stage Ⅰ or Ⅱ disease had better loco-regional control than those with stage ⅡE (P = 0. 03). Patients with primary tumor 8 cm or less in diameter had better loco-regional control than those with primary tumor more than 8 cm in diameter(P =0.01). Patients who un-derwent complete resection had better loco-regional control than those who underwent incomplete resection or no surgery(P=0.03). Patients with stage Ⅰ and Ⅱ disease treated with surgery had more local recurrence, and patients treated without surgery tended to recur systematically. Patients with stage ⅡE disease tended to recur locally in spite of surgery or not. Conclusions The efficacy of surgical and non-surgical treatment for primary gastric MALT lymphoma are similar. Surgical resection is no longer a necessary approach in the primary treatment. Clinical stage is an important prognostic factor for primary gastric MALT lymphoma.