1.Gradient pH co-precipitation to recover proteins from plasma discarded in artificial-liver treatment
Bing XU ; Chen PAN ; Xiao-Zhi JIANG ; Man-Zhi LIN ; Min CHEN ; Zhuo LIN ; Yi CHEN ; Hui-Cong CHEN ;
Chinese Journal of Infectious Diseases 2001;0(05):-
Objective To set up a system in vitro to rapidly recover plasma proteins lost during artificial-liver treatment.Methods The polyprotein precipitation was obtained by all proteins whose isoelectric point pH value were between 7.3 and 5.1,which collided with each other and aggregated using gradient pH co-precipitation(adding 1 mol/L citric acid slowly in the plasma solution to change the pH values gradually from 7.3 to 5.1 in 5 h)combined with salting out(degree of saturation of NaCl is 33%,reacted for 5.5 h at 4℃)or low-temperature ethanol precipitation(40% ethanol, reacted for 5.5 h at -7℃)so that to get rid of toxicants by discarding the supernatant.Results In the range of pH 5.1-7.3,50%(29g/57g)of the total plasma proteins had been recovered by the gradient pH salting out and 41%(25 g/61g)by the gradient pH low-temperature ethanol co-precipi- tation.The protein remained in the supernatant was mostly albumin and its combined bilirubin.The levels of total bilirubin decreased to 0.07% and 0.06% of the original levels by these two methods respectively and the serum HBV DNA level decreased to be undetected(quantitative PCR).Conclu- sions The proteins with close isoelectric point can co-precipitated with the presence of high concen- tration of NaCl or low-temperature ethanol and by changing the pH value gradually.The total protein in the discarded plasma during artificial-liver treatment can be recovered rapidly using the gradient pH coprecipitation.
2.Changes of insulin like growth factor - 1 in blood and cerebrospinal fluid in children with viral encephalitis
yi gang, MAN ; xiao xia, LIN ; zhi-rong, SHU ; zi-pu, LI
Journal of Applied Clinical Pediatrics 2004;0(12):-
Objective To study the changes of insulin- like growth factor- 1(IGF-1)in blood and cerebrospinal fluid (CSF) in children with viral encephalitis (VE).Methods The IGF-1 levels in blood and CSF were determined before treatment by ELISA in 25 children who admitted with VE, including 15 cases with severe VE and another 10 cases with mild VE, 10 children served as con-trols. Results Before treatment, the blood IGF-1 levels in VE group were significantly lower than those of controls, but the CSF IGF-1 levels were significantly higher than those of controls(P0.05), but the blood IGF-1 levels in serve VE group were significanfly lower than those of mild VE group and controls(P
3.Significance of Biological Rhythm on Selective Nocte Treatment on Autosomal Dominant Nocturnal Frontal Lobe Epilepsy
jia-sheng, HU ; zhi-sheng, LIU ; fang-lin, WANG ; xiao-man, WU
Journal of Applied Clinical Pediatrics 2004;0(12):-
0.05).The adverse effect of treatment group was significantly less than control group(P
4.Expression of a testis-specific gene 1700001022RIK in mice and its bioinformatic analysis.
Yu-chi LI ; Shou-ren LIN ; Man-ling LUO ; Huan GUO ; Han-wei WU ; Zhi-mao JIANG ; Yao-ting GUI
National Journal of Andrology 2015;21(5):391-395
OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.
METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.
RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.
CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.
Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis
5.Vascular endothelial growth factor and pigment epithelium-derived factor in aqueous humor of patients with choroidal neovascularization.
Jian-ping TONG ; Ye SHEN ; Wai-man CHAN ; Shun-chao LIN ; Zhi-pei PENG
Journal of Zhejiang University. Medical sciences 2006;35(3):311-314
OBJECTIVETo detect the levels of vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) in aqueous humor of patients with active choroidal neovascularization (CNV).
METHODSAqueous humor samples were obtained from 32 patients with active CNV. The concentrations of VEGF and PEDF in aqueous humor were measured by enzyme linked immunosorbent assay (ELISA) for quantitative analysis. VEGF and PEDF in 10 samples of aqueous humor from patients with cataract were also detected by the same methods as control.
RESULTThe mean VEGF and PEDF concentrations in aqueous humor of active CNV patients were higher than those in the control group (P=0.000).
CONCLUSIONThe patients with active CNV exhibit significantly higher VEGF and PEDF levels than those in control, indicating that VEGF along with PEDF may modulate the formation of CNV.
Adult ; Aged ; Aged, 80 and over ; Aqueous Humor ; chemistry ; Choroidal Neovascularization ; metabolism ; Eye Proteins ; analysis ; Female ; Humans ; Male ; Middle Aged ; Nerve Growth Factors ; analysis ; Serpins ; analysis ; Vascular Endothelial Growth Factor A ; analysis
6.Studies on characteristics of nanostructure of N-TiO2 thin films and photo-bactericidal action.
Ming-fang XU ; Shan LIN ; Xin-man CHEN ; Yan-zhi PENG
Journal of Zhejiang University. Science. B 2006;7(7):586-590
Pseudomonas aeruginosa strain (AS1.50) and Bacillus subtilis strain (AS1.439) from Ming lake were decomposed by photocatalytic nanostructure N-TiO(2) thin films in a photo-reactor under UV irradiation. The different thickness nanostructure N-TiO(2) thin films coated on mesh grid were prepared by sol-gel method and immobilized at 500 degrees C (films A) or 350 degrees C (films B) for 1 h in a muffle furnace. The results showed that N-TiO(2) thin film B (8.18 nm thickness, 2.760 nm height and 25.15 nm diameter) has more uniform granular nanostructure and thinner flat texture than N-TiO(2) thin film A (12.17 nm thickness, 3.578 nm height and 27.50 nm diameter). The bactericidal action of N-TiO(2) thin film A and film B for Pseudomonas aeruginosa strain (AS1.50) and Bacillus subtilis varniger strain (AS1.439) were investigated in this work. More than 95% of photocatalytic bactericidal efficiency for Pseudomonas aeruginosa strain (AS1.50) and 75% for Bacillus subtilis strain (AS1.439) were achieved by using N-TiO(2) thin films-B for 70-80 min of irradiation during the photo-bactericidal experimental process. The results indicated that the photo-induced bactericidal efficiency of N-TiO(2) thin films probably depended on the characteristics of the films.
Anti-Bacterial Agents
;
chemistry
;
pharmacology
;
Bacillus subtilis
;
drug effects
;
radiation effects
;
Membranes, Artificial
;
Nanostructures
;
chemistry
;
ultrastructure
;
Nitrogen Compounds
;
chemistry
;
pharmacology
;
Particle Size
;
Photochemistry
;
methods
;
Pseudomonas aeruginosa
;
drug effects
;
radiation effects
;
Surface Properties
;
Titanium
;
chemistry
;
pharmacology
;
Ultraviolet Rays
7.Overexpression of centromere protein H is significantly associated with breast cancer progression and overall patient survival.
Wen-Ting LIAO ; Yan FENG ; Men-Lin LI ; Guang-Lin LIU ; Man-Zhi LI ; Mu-Sheng ZENG ; Li-Bing SONG
Chinese Journal of Cancer 2011;30(9):627-637
Breast cancer is one of the leading causes of cancer death worldwide. This study aimed to analyze the expression of centromere protein H (CENP-H) in breast cancer and to correlate it with clinicopathologic data, including patient survival. Using reverse transcription-polymerase chain reaction and Western blotting to detect the expression of CENP-H in normal mammary epithelial cells, immortalized mammary epithelial cell lines, and breast cancer cell lines, we observed that the mRNA and protein levels of CENP-H were higher in breast cancer cell lines and in immortalized mammary epithelial cells than in normal mammary epithelial cells. We next examined CENP-H expression in 307 paraffin-embedded archived samples of clinicopathologically characterized breast cancer using immunohistochemistry, and detected high CENP-H expression in 134 (43.6%) samples. Statistical analysis showed that CENP-H expression was related with clinical stage (P = 0.001), T classification (P = 0.032), N classification (P = 0.018), and Ki-67 (P < 0.001). Patients with high CENP-H expression had short overall survival. Multivariate analysis showed that CENP-H expression was an independent prognostic indicator for patient survival. Our results suggest that CENP-H protein is a valuable marker of breast cancer progression and prognosis.
Adult
;
Aged
;
Aged, 80 and over
;
Blotting, Western
;
Breast
;
cytology
;
Breast Neoplasms
;
genetics
;
metabolism
;
pathology
;
Cell Line
;
Cell Line, Tumor
;
Chromosomal Proteins, Non-Histone
;
genetics
;
metabolism
;
Epithelial Cells
;
cytology
;
metabolism
;
Female
;
Follow-Up Studies
;
Humans
;
Ki-67 Antigen
;
metabolism
;
Middle Aged
;
Neoplasm Metastasis
;
Neoplasm Staging
;
RNA, Messenger
;
metabolism
;
Reverse Transcriptase Polymerase Chain Reaction
;
Survival Rate
;
Up-Regulation
8.The Effecacy and Safety of Daratumumab Based Regimens in Relapsed/Refractory Multiple Myeloma:A Single-Center Real-World Data Analysis
Han-Yan ZENG ; Zhi-Juan LIN ; Zhi-Feng LI ; Long LIU ; Man-Man DENG ; Bing XU
Journal of Experimental Hematology 2024;32(3):763-767
Objective:To investigate the efficacy and safety of daratumumab based regimens in relapse and/or refractory multiple myeloma(RRMM)in the real world,as well as the impact of daratumumab on stem cell collection and engraftment.Methods:The clinical data of patients with RRMM who received daratumumab in hematology department of the First Affiliated Hospital of Xiamen University from February 2019 to March 2023 and had evaluable efficacy were retrospective analysis.Results:All 43 RRMM patients were treated with daratumumab-based combination regimens,including Dd,DVd,DRd,Dkd,DId,and Dara-DECP.With median follow-up time 10.1(2.1-36.6)months,the best overall response rate(ORR)was 74.4%and a best complete response rate(CR)was 25.6%.1-year overall survival rate(OS)was 84.5%.The most common severe hematologic adverse events(Grade>3)are 3/4 grade leukopenia(18.6%),and the most common severe non-hematologic adverse events were infusion-related reactions(IRRs,20.9%)and infections(7.0%).Multivariate prognostic analysis showed that extramedullary infiltration was an independent adverse prognostic factor affecting OS(P=0.004).The use of daratumumab has no effect on stem cell collection,or engraftment.Conclusion:Daratumumab is safe and effective in RRMM.
9.Effects of simvastatin on PI3K/AKT signaling pathway in human acute monocytic leukemia cell line SHI-1.
Mei ZENG ; Wei-Ying GU ; Ting-Xiu JIANG ; Zi-Xing CHEN ; Guo-Qiang QIU ; Man LI ; Hao-Qing WU ; Zhi-Lin WANG ; Xiao-Bao XIE ; Xiang-Shan CAO
Journal of Experimental Hematology 2012;20(2):268-272
To investigate the effects of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor simvastatin (SV) on proliferation, apoptosis and the PI3K/AKT signaling pathway in human acute monocytic leukemia cell line SHI-1. SHI-1 cells were incubated with different concentrations of SV (5, 10, 15 µmol/L). Otherwise, SHI-1 cells without any treatment were used as control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detection. MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the early stage apoptosis ratio. The human PI3K-AKT Signaling Pathway RT(2) Profiler(TM) PCR Array was used to detect the expression of 84 genes involved in PI3K-AKT signaling. The results indicated that the SV inhibited the proliferation and inducted the apoptosis of SHI-1 cells in time- and dose-dependent manners significantly. The growth inhibition rates of SHI-1 cells treated with 15 µmol/L SV for 24, 48 and 72 h were 26.82, 47.09 and 63.92, respectively; and their early stage apoptosis ratios were 5.75, 13.25 and 15.59, respectively. Compared with the control group, expression levels of 39 genes were changed in the group of 15 µmol/L SV at 48 h, among them 26 genes were down-regulated and 13 genes were up-regulated. It is concluded that the SV can inhibit proliferation and induce apoptosis of SHI-1 cells, and the mechanism may be associated with the changes of gene expression level in PI3K-AKT signaling pathway regulated by SV.
Apoptosis
;
drug effects
;
Cell Line, Tumor
;
Cell Proliferation
;
drug effects
;
Down-Regulation
;
Gene Expression Regulation, Leukemic
;
Humans
;
Leukemia, Monocytic, Acute
;
metabolism
;
Phosphatidylinositol 3-Kinases
;
metabolism
;
Proto-Oncogene Proteins c-akt
;
metabolism
;
Signal Transduction
;
drug effects
;
Simvastatin
;
pharmacology
10.Effect of estrogen on gene expression of fatty acid synthase in periosteum.
Rui-min ZHENG ; Shou-qing LIN ; Yong LIU ; Man-ting HUANG ; Wei-yan GONG ; Zhi-hong WU
Chinese Medical Journal 2009;122(15):1775-1779
BACKGROUNDEstrogen deficiency contributes to postmenopausal osteoporosis. Periosteum might be a potential target of estrogen, but the underlying mechanism at gene level is far from being elucidated. The objective of this study was to investigate the correlation between estrogen and fatty acid synthase (FAS) expression in periosteum.
METHODSHuman periosteum cells were cultured in vitro. Expressed genes in the substrated cDNA library were verified using semi-quantitative PCR and real-time PCR. The expression of FAS in periosteum of ovarectomized (OVX) SD rats was investigated.
RESULTSFAS gene was most significantly expressed in the subtracted cDNA library of periosteal cells screened by semi-quantitative PCR. Low FAS expression was verified by real-time PCR in the estrogen exposed human periosteum rather than in the control. The estradiol levels were (20.81 +/- 12.62) pg/ml, (19.64 +/- 4.35) pg/ml and (13.47 +/- 1.84) pg/ml in the sham group, the control, and the OVX group, respectively. The estradiol levels in the OVX group was significantly lower (P = 0.0386). The FAS gene expression in periosteum in the OVX group, sham group, and control group was 3.09 +/- 1.97, 1.33 +/- 0.47 and 1.51 +/- 1.32, respectively. The gene expression in the OVX group was significantly higher (P = 0.0372).
CONCLUSIONEstrogen modulates FAS gene expression in in vitro human perisoteum as well as in in vivo rat periosteum.
Animals ; Cells, Cultured ; Estradiol ; blood ; pharmacology ; physiology ; Fatty Acid Synthases ; genetics ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Ovariectomy ; Periosteum ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction