2.Development of monoclonal antibodies against PD-L1 and preliminary investigation on potential application in treatment of chronic hepatitis B virus in-fection
Yong WU ; Tianying ZHANG ; Liuwei SONG ; Ningshao XIA ; Quan YUAN
Chinese Journal of Immunology 2016;32(7):1004-1008,1012
Objective:To get specific monoclonal antibodies ( mAbs) against PD-L1 which can block PD-1/PD-L1 binding, and explore the feasibility of its application on the treatment of chronic HBV infection preliminarily by in vitro and in vivo model. Methods:E. coli expression and series chromatography purification system were employed to get human and mouse PD-1/PD-L1 that had binding activity in vitro. By immunizing BALB/c mouse with purified recombination proteins of PD-L1,mAb hybridoma cell lines against PD-L1 were obtained. The reactivity with human/mice PD-L1 of individual antibody and the interaction blocking activity of the mAbs to PD-1/PD-L1 in vitro were examined by indirect chemiluminescence immune assay. Results: 8 cell lines against PD-L1 were obtained and 2 Anti-PDL1 mAbs (Ab5 &Ab6) performed strong immune activity to human/mice PD-L1 and blocking activity to PD-1/PD-L1. In the PBMC stimulation experiment of chronic HBV patient,Ab5 and Ab6 could promote theγ-IFN levels. With HBV in-fecting mice model,intravenous injections of these mAbs induced dramatically decrease of HBV DNA copies about 20 times, HBsAg levels in serum reduced to 30% of the baseline level. Conclusion:We obtained 2 PD-L1 mAbs with the reactivity to human/mice PD-L1 and blocking activity to PD-1/PD-L1. The 2 mAbs can promote T cell function in PBMC stimulation culture of chronic HBV patient, have significant antiviral effect in HBV transgenic mice and can be candidates for immunotherapy applications.
3.Chemical of Vitex trifolia.
Quan-Yu LIU ; Yong-Sheng CHEN ; Fei WANG ; Shi-Wu CHEN ; Yong-Hong ZHANG
China Journal of Chinese Materia Medica 2014;39(11):2024-2028
A new steroidal ester, beta-rosaterol palmitate (1) along with ten known compounds, uvaol(2), 3-epi-ursolic acid (3), 2alpha, 3beta, 24-trihydroxyolean-12-en-28-oic acid (4), 2alpha, 3alpha, 24-trihydroxyurs-12-en-28-oic acid (5), 2alpha, 3alpha, 24-trihydroxyolean-12-en-28-oic acid (6), 2alpha, 3alpha, 24-trihydroxyolean-12-en-28-oic acid-28-O-beta-D-glucopyranosyl ester (7), (Z)-9-hexadecenoic acid (8), octacosyl alcohol (9), beta-sitosterol (10) and beta-daucosterol (11), has been isolated from the stems and leaves of Vitex trifolia. Their structures were elucidated using a combination of 1D and 2D NMR techniques (COSY, HMQC, and HMBC)and HR-ESI-MS analyses. Compounds 2-7 were isolated from this plant for the first time.
Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Magnetic Resonance Spectroscopy
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Molecular Structure
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Spectrometry, Mass, Electrospray Ionization
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Vitex
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chemistry
4.Clinical analysis of 126 children with hemorrhagic fever with renal syndrome.
Cui-ping WU ; Ru-yong LI ; Liang-yi QI ; De-quan XIA
Chinese Journal of Pediatrics 2003;41(9):703-704
Adolescent
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Child
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Child, Preschool
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Electrocardiography
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Female
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Fever
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complications
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Hemorrhagic Fever with Renal Syndrome
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blood
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complications
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pathology
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Humans
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Hypergammaglobulinemia
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blood
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Immunoglobulin M
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blood
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Male
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Pain
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complications
5.A fMRI study of item memory and source memory
Ming-Quan WANG ; Kai WANG ; Yong-Qiang YU ; Hai-Bao WANG ; Cheng ZHANG ; Jin-Min WU ;
Chinese Journal of Neurology 2000;0(05):-
Objective To examine the neural basis of item memory and source memory with fMRI approach.Methods Eight male and eight female healthy fight-handed native Chinese speakers were involved in this study.The item memory and source memory task were conducted with 504 highly frequent Chinese double-character words in the Block-designed experiment.Participants underwent such a double- round procedure as fMRI scanning following study.The fMRI data collected from a GE 1.5T MRI system were analyzed to generate corresponding activation maps for females and males respectively(P20)using statistical parametric mapping software(SPM).Results For females,item memory task activated the bilateral dorsolateral prefrontal cortex(BA6,the number of activated voxel clusters was 62 or 11 in the left and the right,respectively),source memory more activated the left dorsolateral prefrontal cortex(BA6/46,the number was 59).For males,item memory activated the right dorsolateral prefrontal cortex(BA6/46,the number was 64),source memory activated the bilateral dorsolateral prefrontal cortex(BA6,9 and 40 in the left and the right).Conclusion On the neural basis of item or source memory,there exists dissociation,which is that right dorsolateral prefrontal areas are more activated by item memory while left dorsolateral prefrontal areas by source memory.For the difference of gender,it is suggested that left dorsolateral prefrontal areas(BA6/46)are more activated in females while right dorsolateral prefrontal cortex(BA6/46)more in males.
6.Dose-respones relationship between daily total fluoride intake and prevalence of osteofluorosis
Quan-yong, XIANG ; Minh-hao, ZHOU ; Ming, WU ; Ran, TAO ; Lian-sheng, CHEN ; Ming-fang, ZHANG ; You-xin, LIANG
Chinese Journal of Endemiology 2008;27(2):196-200
Objective To investigate the daily total fluoride intake in relating to the prevalence of skeletal fluorosis in two villages in Jiangsu Province,in order to provide the scientific evidences for the control and prevention of endemic fluorosis.Methods Adults sampled from a high-fluoride Village,Wamiao,and a low-fluoride Village,Xinhuai,were surveyed in this study according to the fluoride concentration in their household shallow well.The average daily total fluoride intake from difierent sources and the skeletal fluorosis were investigated in each subject.Then the subjects from two villages were allocated into five subgroups(<2.00,2.00~,3.00~,4.00~,≥5.00 mg/d),the relation fluoride intake and prevalence of osteofluorosis was analyzed.Results The prevalence of skeletal fluorosis in Wamiao Village was 31.06%(41/132),but no skeletal fluorosis case(0/35)was found in Xinhuai Village.According to the daily total fluoride intake,subjects with higher daily total fluoride intake tended to associated with a higher prevalence of skeletal fluorosis in a significant dose-response relationship(regression equation:y=2.624-6.855x+3.424x2:r=0.997).The benchmark dose lower limitation of daily total fluoride intake with 95% confidencewas 2.50 mg/d calculated according to this dose-response relationship,the reference dose(RfD)was 2.50 mg/d.In Wamiao Village a significant difference was also found between daily total fluoride intake in 41 subjects[(5.09±1.20)mg/d]with X-ray detectable skeletal fluorosis and in 91 subjects[(3.08±1.12)mg/d]without X-ray detectable skeletal fluorosis(t=-9.32,P<0.01).Conclusions These findings indicate that the daily total fluoride intake has a significant dose-response relationship with the prevalence of skeletal fluorosis in an endemic fluorosis area associated with high-fluoride drinking water;and the RfD in this study was lower than that in the national standard of"Chinese hygienic standard for daily total fluoride intake(WS/T 87-1996)"(3.50 mg/d).
7.Retrospective study on post-operative glucose level and insulin dose in patients undergoing total pancre-atectomy
Ying ZHOU ; Weigang ZHAO ; Wenming WU ; Tao YUAN ; Yong FU ; Taiping ZHANG ; Menghua DAI ; Xin LU ; Quan LIAO ; Yupei ZHAO
Chinese Journal of Clinical Nutrition 2016;24(2):70-75
Objective To evaluate the post-operative glucose level and insulin dose of patients undergoing total pancreatectomy.Methods From September 1980 to September 2014, 21 patients underwent total pancrea-tectomy in Peking Union Medical College Hospital, who were enrolled in our study.We reviewed the changes in their insulin dosage and glucose levels after operation, also summarized type and dose of insulin as well as glucose level in stable period.Results The required insulin dose reached peak within 4 days after surgery ( maximum dose 300 U/d).The average dose was (143.5 ±62.8) U/d and decreased gradually.During the perioperative period (needing parenteral nutrition), the blood glucose level fluctuated markedly (1.52-29.06 mmol/L) and the average level was (11.18 ±0.95) mmol/L.During the stable period ( without parenteral nutrition) , patients on average had (5.3 ±2.0) U of preprandial rapid-acting insulin and (8.1 ±2.9) U of long-acting insulin be-fore sleeping;the average fasting blood glucose was (6.69 ±1.48) mmol/L, 2 h postprandial blood glucose was (9.08 ±2.84) mmol/L, bedtime blood glucose was (9.66 ±2.49) mmol/L, and blood glucose level at night was (8.15 ±2.78) mmol/L.67%of the patients had 13 hypoglycemic episodes monthly on average.For those five followed-up patients, the average hemoglobin A1c was (6.15 ±1.20)%.Conclusions Patients undergoing total pancreatectomy may experience marked fluctuation of blood glucose level and short-term increase of insulin need which gradually decreases afterwards.After entering the stable period, the glucose level could be well-con-trolled but with frequent hypoglycemia.There is no diabetic ketoacidosis.
8.Spatio-temporal expression of the pathway-specific regulatory gene redD in S. coelicolor.
Li-hua ZHOU ; Yu-qin LI ; Yong-quan LI ; Dan WU
Journal of Zhejiang University. Science. B 2005;6(6):464-469
Confocal laser scanning microscopy was used to observe the spatio-temporal expression of the pathway-specific gene redD during S. coelicolor cell cultivation. The corresponding mutant S. coelicolor lyqRY1522 carrying redD::eyfp in the chromosome was constructed. The temporal expression results of the fusion protein during submerged cultivation demonstrated that expression of redD began in the transition phase, continuing through the exponential growth phase to the stationary phase, and reached maximum in the stationary phase. On the other hand, redD was expressed only in substrate mycelia during solid-state culture, while aerial mycelia remained essentially non-fluorescent throughout culture. Results demonstrated that the expression pattern of redD coincides with that of the biosynthesis of the antibiotics during culture, revealing a direct correlation between the spatio-temporal distribution of regulatory gene expression and second metabolism.
Anti-Bacterial Agents
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biosynthesis
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Bacterial Proteins
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genetics
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metabolism
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Gene Expression Profiling
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Gene Expression Regulation, Bacterial
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physiology
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Mutation
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Signal Transduction
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physiology
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Streptomyces coelicolor
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genetics
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metabolism
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Trans-Activators
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genetics
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metabolism
9.Effect of ginsenoside Rg1 on the spatial learning-memory ability in dementia rats after transplanted with bone marrow mesenchymal stem cells.
Wei WU ; Jing-quan YANG ; Zhi-yong HE
Chinese Journal of Integrated Traditional and Western Medicine 2011;31(6):799-802
OBJECTIVETo study the effect and mechanism of ginsenoside Rg1 on the spatial learning-memory ability in rats with Alzheimer's disease after transplanted with bone marrow mesenchymal stem cells (BMSCs).
METHODSUsing digital randomization table method, seventy-five male SD rats were divided into the bilateral FF transection model group (as the model group: ambi-hippocampal fimbria-fomix transected), the sham-operative control group (the SOC group: receiving the same modeling process as the model group, but without ambi-hippocampal fimbria-fomix transected), the ginsenoside Rg1 treatment group (as the treatment group: Two weeks after modeling ginsenoside Rg1 was peritoneally injected at the dose of 5 mg/kg, once daily for four weeks in total), the BMSCs transplanted treatment group [as the control group: Two weeks after modeling every rat received transplantation of BMSCs (10 microL, 1 x 10(6) cells)], and the ginsenoside Rg1 + BMSCs treatment group (as the combination group: They received both transplantation of BMSCs and peritoneal injection of ginsenoside Rgl). The spatial learning-memory ability of rats was detected by Morris water maze and the escape latency (s) was recorded, mRNA expression of nerve growth factor (NGF) was detected using Real-time PCR.
RESULTSSix weeks after the hippocampal fimbria-fomix (FF) transection, the escape latency o feach medication group was obviously shorter than that of the model group, and the spatial learning-memory ability of dementia rats was somewhat improved. The spatial learning-memory ability of rats in the combination group was (29.95 +/- 2.03) and the mRNA expression level of NGF was (1.13 +/- 0.15), better than those in the BMSCs group (44.36 +/- 1.43, 0.78 +/- 0.09, P<0.05).
CONCLUSIONSGinsenoside Rg1 could strengthen the spatial learning-memory ability in dementia rats after transplanted with BMSCs. Its mechanism might be possibly correlated with up-regulating mRNA expression of NGF in basal forebrain after BMSCs transplantation.
Animals ; Dementia ; psychology ; therapy ; Ginsenosides ; pharmacology ; therapeutic use ; Learning ; drug effects ; Male ; Memory ; drug effects ; Mesenchymal Stem Cell Transplantation ; Rats ; Rats, Sprague-Dawley
10.Isolation,incubation and identification of parenchymal neural stem cells in adult mouse spinal cord
Hui ZHANG ; Zong-Sheng YIN ; Sheng-Quan ZHANG ; Wei WANG ; Xing-Yi HUA ; Yong HU ; Guang-Wu LI ;
Chinese Journal of Trauma 2003;0(07):-
Objective To isolate and identify the adult neural stem cells from the parenchyma of spinal cord in adult mouse.Methods The parenchymal spinal cord from adult mouse was dissected and dissociated by mechanical trituration.The tissue suspension was cultured in serum-free DMEM/F12 medium supplemented with EGF and B27.The cell colonies generated from a single cell were screened by limited dilution and incubated with BrdU.The cell colonies were transferred into medium with serum to induce differentiation.The cells were identified with antibodies to Nestin,BrdU,MAP2 and GFAP by immunofluorescence staining.Results The cells were cultured for seven days to generate proliferative neurospheres.The majority of cells in these neurospheres expressed Nestin and were differentiated into MAP2-positive cells and GFAP-positive cells in medium containing with fetal bovine serum.Conclusion A significant number of neural stem cells are present in the parenchymal adult mouse spinal cord and can proliferate and also give rise to neurons and glia in vitro.