1.Cytocompatibility of biodegradable poly(ethylene glycol)-b-poly(L-lactide)-b-poly(L-glutamic acid) tri-block copolymer with human umbilical vein endothelial cells
Bin LIU ; Xianjing SONG ; Jieyu LIU ; Feng JIANG ; Yongfeng SHI ; Dongsi SHUANG ; Zhibo LI
Chinese Journal of Tissue Engineering Research 2008;12(10):1950-1953
BACKGROUND: Poly(ethylene glycol)-b-poly(L-lactide)-b-poly(L-glutamic acid) (PEG-PLA-PGL) tri-block copolymers have good applied foreground in constructing tissue engineering scaffold materials. Whether endothelial cells survive and grow on the materials has a direct influence on the application as a biodegradable material for the scaffold of endothelial cell vector.OBJECTIVE: To explore the cytocompatibility of PEG-PLA-PGL tri-block copolymers with human umbilical vein endothelial cells (HUVECs).DESIGN: Randomized control observation.SETTING: the Second Hospital of Jilin University.MATERIALS: The experiment was carried out in the Department of Pathobiology, School of Basic Medical Sciences, Jilin University from February to October in 2006. Human umbilical cord about 20 cm length came from one neonatal infant who was delivered normally after enough months in the Department of Gynecology and Obstetrics, the Second Hospital of Jilin University. Human umbilical cord was sampled in the informed consents of the infant's family member. The experimentation was authorized by the medical ethic committee of the hospital. PEG-PLA-PGL membranes were provided by Changchun Institute of Applied Chemistry, Chinese Academy of Sciences. Inverted microscope and phase-contrast microscope were bought from Olympus Company (Japan).METHODS: HUVECs cultivated and grew steadily, were inoculated onto PEG-PLA-PGL membranes, serving as the experiment group. While the culture medium without PEG-PLA-PGL membranes were taken as the control group.①Cytocompatibility of PEG-PLA-PGL membranes with HUVECs was evaluated by observing cellular growth through phase-contrast microscope.②The proliferation index of cells was detected by MTT method in 1, 3, 5 and 7 days after inoculation.MAIN OUTCOME MEASURES: ①Cytocompatibility of PEG-PLA-PGL membranes with HUVECs;②The proliferation index of cells in l, 3, 5 and 7 days after inoculationRESULTS: ①Cytocompatibility of PEG-PLA-PGL membranes with HUVECs: The observation result of phase contrast microscopy showed that, endothelial cells planted on the PEG-PLA-PGL membranes began to attach and stretch after being planted 4-6 hours. Three days later, cells grew in colonies rapidly, after 5 days, colonies began to fuse and seemed like cobble-stone. The cells were shuttle or polygon in shape after passages. There were no significant differences between the experiment and control group. Cells cultured on PEG-PLA-PGL membranes for 15 days grew in inserts with membranes, but they didn't grow into patches through scanning electron microscope.②The proliferation index of cells: No significant differences of the proliferation index of cells were detected by MTT method in 1, 3, 5 and 7 days after inoculation between experiment group and control group (P>0.05).CONCLUSION: Endothelial cells grow well in PEG-PLA-PGL membranes, and the two have good cytocompatibility.
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. Peripheral nerve injury in LAMA2-related congenital muscular dystrophy patients
Xiaoping LIANG ; Shuang WANG ; Wei ZHANG ; Yun YUAN ; Juan DING ; Xingzhi CHANG ; Cuijie WEI ; Jieyu LIU ; Hui XIONG
Chinese Journal of Pediatrics 2017;55(2):95-99
Objective:
To explore the injury pattern and features of peripheral nerve in congenital muscular dystrophy patients caused by LAMA2 gene mutation.
Method:
Seventeen patients genetically or molecular pathologically diagnosed as LAMA2-related congenital muscular dystrophy were recruited in Peking University First Hospital between 2002 and 2015. All the patients received nerve conduction velocity (NCV) and needle electromyography tests. Clinical and laboratory examination data of the patients was retrospectively analyzed. The correlation between the NCV and disease course was determined by Pearson correlation analysis. Additionally, one patient underwent a sural nerve biopsy.
Result:
Among these 17 identified patients (13 male and 4 female), all of them were diagnosed as congenital muscular dystrophy, and all of them underwent electrophysiological examination at ages between 1 month to 6 years. Electromyogram indicated seventeen patients of myogenic damage, of whom 10 cases were complicated with reduced NCV. Twenty-six of 95 analyzed nerves showed NCV slower than the normal average of contemporary in 17%-47%. Correlation analysis between NCV and the disease course indicated that NCV of median nerves, ulnar nerves, tibial nerves and common peroneal nerves were negatively associated with the disease course (