1.Property and implication of dendritic cells from peripheral blood monocytes in patients with persistent hepatitis B virus infection.
Lihe XING ; Fusheng WANG ; Mingxu LIU
Chinese Journal of Infectious Diseases 2001;0(06):-
Objective To identify the property and implications of dendritic cells of peripheral blood monocytes (PBMC) in patients with persistent HBV infection. Methods The PBMC from 18 patients with persistent HBV infection and 10 healthy subjects were incubated and induced into mature dendritic cells (DC) in the completed medium containing GM CSF、IL 4、FLt3 L and TNF ? cytokines. The flow cytometric analysis was employed to detect the expression of surface markers on DC. ELISA test was used to determine the level of interleukin 12 (IL 12) and IL 10 cytokines produced by DCs. The stimulatory capacity of DC were also determined in allogenic mixed leukocyte reaction (MLR). Results A typical morphology of DCs was observed when the PBMCs from healthy subjects and HBV infected patients were induced for 6 7 day in vitro incubation, but there were some significant differences of DCs as follows: (1) the proliferation ability and number of DCs significantly decreased, in particular, the expression levels of HLA DR,CD80(B7 1),CD86(B7 2) and CD1? on DC surface were simultaneously much lower in patients compared to healthy subjects ( P
2.Design and synthesis of photoaffinity biotin labelled 2'-O-propargyl-guanosine.
Luxin NA ; Xin LIU ; Zhuoming MENG ; Zhu GUAN ; Lihe ZHANG ; Zhenjun YANG
Acta Pharmaceutica Sinica 2015;50(1):59-63
Photoaffinity labeling is widely applied to demonstrate targets of small molecule ligands. In this paper, biotin photoaffinity labeled molecule with propargyl group 1 has been designed and synthesized, followed it's labeling of N2-acetyl-2'-O-propargyl guanosine 9 by "click chemistry". This technology presents delight development potential in labeling of second messenger cyclic nucleotide, antisense oligonucleotide or siRNA.
3.Long-term culture and differentiation of skin-derived mesenchymal stem cells.
Liye YANG ; Xiangming LIU ; Guozhen HUI ; Jian FEI ; Lihe GUO
Journal of Biomedical Engineering 2005;22(3):514-517
The aim of this study was to investigate the culture conditions of skin-derived mesenchymal stem cell (sMSCs) and to explore a new cell source for central nervous system cell transplantation. The cells from skins of mice were primarily isolated and cultured in serum-free medium, and they were transferred into serum-containing medium after passaged 2, and the passaged cells were identified by immunocytochemistry and induced to differentiate into multiple lineages. The results indicated that a population of sMSCs could be isolated from skins, they could be maintained in vitro for extended periods with stable population doubling, and they were expanded as undifferentiated cells in culture for more than 10 passages, indicating their proliferative capacity. About 60% of sMSCs expressed vimentin and the majorities of these cells expressed fibronectin. They could differentiate into adipocytes, osteogenic cells and fibroblast-like cells, they could differentiate into neurons with a simple protocol, and almost 50-60% of these cells expressed neuron specific enolase (NSE) and neurofilament (NF); and the differentiated neurons showed typical complicated morphology of neurons. In conclusion, skin contains stem cells that are capable of multiple differentiation; they could be cultured in vitro for long time and could maintain their characteristics of stem cells, and they may represent an alternative autologous stem cell source for CNS cell transplantation.
Adipocytes
;
cytology
;
Animals
;
Cell Culture Techniques
;
Cell Differentiation
;
Cells, Cultured
;
Culture Media, Serum-Free
;
Fibroblasts
;
cytology
;
Immunohistochemistry
;
Mesenchymal Stromal Cells
;
cytology
;
Mice
;
Neurons
;
cytology
;
Skin
;
cytology
4.A novel homozygous mutation p.E25X in the HSD3B2 gene causing salt wasting 3β-hydroxysteroid dehydrogenases deficiency in a Chinese pubertal girl: a delayed diagnosis until recurrent ovary cysts.
Yonglan HUANG ; Jipeng ZHENG ; Ting XIE ; Qing XIAO ; Shaomei LU ; Xiuzhen LI ; Jing CHENG ; Lihe CHEN ; Li LIU
Chinese Journal of Pediatrics 2014;52(12):948-951
OBJECTIVE3β- hydroxysteroid dehydrogenase deficiency (3βHSD), a rare form of congenital adrenal hyperplasia (CAH) resulted from mutations in the HSD3B2 gene that impair steroidogenesis in both adrenals and gonads. We report clinical features and the results of HSD3B2 gene analysis of a Chinese pubertal girl with salt wasting 3βHSD deficiency.
METHODWe retrospectively reviewed clinical presentations and steroid profiles of the patient diagnosed in Guangzhou Women and Children's Medical Center in 2013. PCR and direct sequencing were used to identify any mutation in the HSD3B2 gene.
RESULTA 13-year-old girl was diagnosed as CAH after birth because of salt-wasting with mild clitorimegaly and then was treated with glucocorticoid replacement. Breast and pubic hair development were normal, and menarche occurred at 12 yr, followed by menstrual bleeding about every 45 days. In the last one year laparoscopic operation and ovariocentesis were performed one after another for recurrent ovary cysts. Under corticoid acetate therapy, ACTH 17.10 pmol/L (normal 0-10.12), testosterone 1.31 nmol/L (normal <0.7), dehydroepiandrosterone sulfate 13.30 µmol/L (normal 0.95 - 11.67), cortisol 720 nmol/L (normal 130-772.8), androstenedione, 17-hydroxyprogesterone and progesterone were normal. Estradiol 461 pmol/L, follicle-stimulating hormone 3.04 IU/L, luteinizing hormone 8.52 IU/L in follicular phase. A pelvic ultrasound showed lateral ovaries cysts (58 mm × 50 mm × 35 mm) and a midcycle-type endometrium. A novel nonsense mutation c.73G >T (p.E25X) was identified in HSD3B2 gene. The girl was homozygous and her mother was heterozygous, while her father was not identified with this mutation.
CONCLUSIONA classic 3βHSD deficiency is characterized by salt wasting and mild virilization in female. Ovary cysts may be the one of features of gonad phenotype indicating ovary 3βHSD deficiency. A novel homozygous mutation c.73G >T(p.E25X) was related to the classical phenotype.
17-alpha-Hydroxyprogesterone ; Adolescent ; Adrenal Hyperplasia, Congenital ; diagnosis ; genetics ; Androstenedione ; China ; Codon, Nonsense ; Delayed Diagnosis ; Female ; Follicle Stimulating Hormone ; Homozygote ; Humans ; Hydrocortisone ; Luteinizing Hormone ; Mutation ; genetics ; Ovarian Cysts ; genetics ; Progesterone Reductase ; genetics ; Recurrence ; Retrospective Studies
5.Clinical characteristics of 34 cases with Japanese encephalitis in adults
Tianhong WANG ; Youquan GU ; Chaoning ZHOU ; Xiaoming CHEN ; Ying WANG ; Lihe YAO ; Wenjuan WU ; Yaqin LU ; Ning LIU ; Jun CHEN
Chinese Journal of Neurology 2018;51(8):612-617
Objective To study the clinical characteristics of Japanese encephalitis (JE) in 34 adult patients and to improve the level of diagnosis of this disease.Methods The clinical manifestations,laboratory results and radiological features of 34 adult patients with JE in our hospital from July 2017 to September 2017 were summarized and the progonsis was observed.The modified Rankin Scale (mRS) was used to evaluate the progonsis.Results Eighteen patients were males and 16 patients were females with the average age of (45.39 ± 16.34) years in 34 patients who were diagnosed as JE.The major clinical features of JE patients included fever (34,100%) with the average temperature of (39.4 ± 1.1) ℃ on admission,headache (26,76%),seizures (7,21%),decreased consciousness (25,74%) on day 2.6 ± 1.4 after the onset,respiratory failure (9,26%) on day 3.8 ± 1.6 after the onset.The major features of laboratory results included white blood cells increase (15,44%),blood hematocrit decrease (25,74%),eosinophil absolute value decrease (29,85%),cerebrospinal fluid pressure increase (12,35%),cerebrospinal fluid protein increase (27,79%),cerebrospinal fluid white blood cells increase (30,88%).Brain MRI scan of abnormal signal was found abnormal in up to 54%patients (14/26),involving the thalamus,basal ganglia,mesencephalon,temporal lobe,hippocampus and occipital lobe,especially in the area of bilateral thalamus and mesencephalon.The follow-up showed three cases were dead;mRS score was 0 in twenty-one cases,1 or 2 in five cases,3 or 4 in three cases,5 in two cases five-six months after onset;the sequelaes were cognitive impairment in nine patients and movement disorder in five patients.Conclusions The clinical symptoms of JE in adults are severe.The main clinical manifestations of JE are hyperthermia,disturbance of consciousness,seizures and respiratory failure,with characteristic imaging findings on brain MRI.JE is a disease with high mortality and severe long-term sequelae.
6.Molecular pathological mechanism of liver metabolic disorder in mice with severe spinal muscular atrophy.
Lihe LIU ; Mingrui ZHU ; Yifan WANG ; Bo WAN ; Zhi JIANG
Journal of Southern Medical University 2023;43(5):852-858
OBJECTIVE:
To explore the molecular pathological mechanism of liver metabolic disorder in severe spinal muscular atrophy (SMA).
METHODS:
The transgenic mice with type Ⅰ SMA (Smn-/- SMN20tg/2tg) and littermate control mice (Smn+/- SMN20tg/2tg) were observed for milk suckling behavior and body weight changes after birth. The mice with type Ⅰ SMA mice were given an intraperitoneal injection of 20% glucose solution or saline (15 μL/12 h), and their survival time was recorded. GO enrichment analysis was performed using the RNA-Seq data of the liver of type Ⅰ SMA and littermate control mice, and the results were verified using quantitative real-time PCR. Bisulfite sequencing was performed to examine CpG island methylation level in Fasn gene promoter region in the liver of the neonatal mice.
RESULTS:
The neonatal mice with type Ⅰ SMA showed normal milk suckling behavior but had lower body weight than the littermate control mice on the second day after birth. Intraperitoneal injection of glucose solution every 12 h significantly improved the median survival time of type Ⅰ SMA mice from 9±1.3 to 11± 1.5 days (P < 0.05). Analysis of the RNA-Seq data of the liver showed that the expression of the target genes of PPARα related to lipid metabolism and mitochondrial β oxidation were down-regulated in the liver of type Ⅰ SMA mice. Type Ⅰ SMA mice had higher methylation level of the Fasn promoter region in the liver than the littermate control mice (76.44% vs 58.67%). In primary cultures of hepatocytes from type Ⅰ SMA mice, treatment with 5-AzaC significantly up-regulated the expressions of the genes related to lipid metabolism by over 1 fold (P < 0.01).
CONCLUSION
Type Ⅰ SMA mice have liver metabolic disorder, and the down-regulation of the target genes of PPARα related to lipid and glucose metabolism due to persistent DNA methylation contributes to the progression of SMA.
Mice
;
Animals
;
PPAR alpha
;
Liver Diseases
;
Muscular Atrophy, Spinal/genetics*
;
Mice, Transgenic
;
Body Weight
;
Glucose