1. Induction of acute myelogenous leukemia cell apoptosis by bortezomib and its effect on SALL4 gene expression
Chinese Pharmaceutical Journal 2012;47(4):274-278
OBJECTIVE: To investigate the proteasome inhibitor bortezomib induces acute myelogenous leukemia cell lines TF1 and NB4 apoptosis and its effect on SALL4 gene expression. METHODS: Cell proliferations were analyzed by MTT assay. Flow cytometry was used to analyze cell apoptosis rate. SALL4 protein was detected by immunocytochemistry. The expressions of SALL4 gene were detected by Real-time PCR. SALL4 proteins in two cell lines were detected by Western Blotting. RESULTS: MTT assay showed bortezomib inhibited the proliferations of two cell lines in a time-and-does manner. TF1 and NB4 cells' 48 h IC50 were (29.15 ± 0.55) and (30.55 ± 0.74) nmol · L-1 respectively. Flow cytometry showed bortezomib could induce apoptosis of two cell lines in a dose-dependent manner. Immunocytochemistry analysis revealed that both of two cell lines expressed SALL4 proteins which located in cell nucleus. Real-time PCR demonstrated that SALL4 genes were down-regulated after cells were treated by different concentrations(10, 30, 50 nmol · L-1) of bortezomib for 24 h, and bortezomib 50 nmol · L-1 groups' genes were down-regulated to 45.11% (TF1) and 69.77% (NB4) respectively comparing with the control groups (P < 0.05). Western blotting revealed that both of the cell lines expressed SALL4B proteins and which could be inhibited by bortezomib in a time-and-does manner. CONCLUSION: Bortezomib can significantly inhibit two cell lines proliferation and induce apoptosis, meanwhile down-regulate the expressions of SALL4 gene. Copyright 2012 by the Chinese Pharmaceutical Association.
2.Four ports pars plana vitrectomy in retrieving dislocated posterior chamber intraocular lens
Lei, GAO ; Fu-Hua, WANG ; Ying, SUN
International Eye Science 2007;7(1):43-45
AIM: To report the use of modified four ports pars plana vitrectomy in managing a dislocated posterior chamber intraocular lens.METHODS: A young man with bilateral pseudophakia and dislocated PC-IOLs had undergone modified four ports pars plana vitrectomy in relocating a dislocated posterior chamber intraocular lens. This procedure was surgically less tedious and demanding compared with previous methods of IOL retrieval.RESULTS: No significant intra- or post-operative complication was noted. The visual recovery of the operated eye was rapid with the best corrected visual acuity at 14 months post-operation being 0.5. In contrast, the right eye without operation due to patient refusal developed total retinal detachment with hand movement visual acuity.CONCLUSION: Modified four ports pars plana vitrectomy may be a more ophthalmic surgeon friendly alternative in the retrieval of dislocated IOL.
4.Expression, purification and characterization of recombinant human type Ⅲ collagen
Xin LIU ; Hua LI ; Lei QI ; Fu SONG ; Lanju XU
International Journal of Biomedical Engineering 2021;44(1):18-22,33
Objective:To express and purify recombinant human collagen type Ⅲ and evaluate its properties.Methods:The recombinant genetic engineering strain pET30a(+)-1880/pACYCDuet-hy726/bL21(DE3) was constructed to stably co-express recombinant human type Ⅲ collagen (rhCol) and prolyl hydroxylase. rhCol was prepared and purified by E. coli high-density fermentation, salting out and column chromatography protein purification technology. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to determine the purity of rhCol. The N-terminal amino acid sequences of rhCol were determined by automatic protein polypeptide sequencing instrument. The hydroxyproline content of rhCol was determined by ultraviolet spectrophotometry. The cellular compatibility of rhCol was evaluated by MTT assay. Results:The final wet weight of high-density fermentation was about 200 g/L. The expression level was about 3 g/L. The purity of rhCol by affinity chromatography was over 95%. The results showed that the hydroxyproline content of rhCol was 11.44%, and the rhCol products have good water solubility and cell compatibility.Conclusions:RhCol can be widely applied to the field of skin care and biomedicine as an excellent biological material.
5.Pharmacological study on free anthraquinones compounds in rhubarb in rats with experimental acute pancreatitis.
Lin ZHU ; Jian-Lei ZHAO ; Xiao-Hang PENG ; Mei-Hua WAN ; Xi HUANG ; Wen-Fu TANG
China Journal of Chinese Materia Medica 2014;39(2):304-308
OBJECTIVETo verify the pharmacological hypothesis of prescriptions by studying the targeted distribution of major components in stewed rhubarb in the rat model with acute pancreatitis (AP).
METHODNormal SD rats (control group, n = 5) and the AP model induced with intraperitoneal cerulein (model group, n = 5) were taken as the experimental objects. Rats of the two groups were orally administered with stewed rhubarb granules (20 g x kg(-1)). Their heart, liver, spleen, lung, kidney and pancreas were collected two hours after the administration. Such constituents as emodin, chrysophanol, physcion, rhein and aloe-emodin and their concentrations in each tissue homogenate were detected by high performance liquid chromatography-mass-mass.
RESULTAloe-emodin and physcion in stewed rhubarb whose concentrations in liver and kidney of normal rats were higher than that in pancreatic tissues, while the distribution spectrums and concentrations of the remaining components in pancreatic tissues had no significant difference with that of other organs. The concentrations of emodin, aloe-emodin, rhein and chrysophanol in stewed rhubarb in pancreatic tissues of the AP model group were higher than that in other tissues and organs, while their concentrations in pancreatic, renal and splenic tissues were notably higher than that in the normal group.
CONCLUSIONIn the conditions of AP, effective components in stewed rhubarb show a targeted distribution feature in pancreas, which provides experimental basis for the pharmacological hypothesis of prescriptions.
Acute Disease ; Animals ; Anthraquinones ; pharmacokinetics ; pharmacology ; therapeutic use ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; therapeutic use ; Male ; Organ Specificity ; Pancreatitis ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry
7.Clinical study of prognosls evaluation of brainstem auditory evoked potential combined with mesencephalic morphology on coma patients with severe craniocerebral injury
Lei WANG ; Fu HUANG ; Qinghua ZHANG ; Zhenlin ZHAO ; Dong HU ; Hua XIAO
Chinese Journal of Postgraduates of Medicine 2012;35(17):1-4
Objective To explore the correlation between brainstem auditory evoked potential (BAEP) combined with mesencephalic morphology and the prognosis of coma patients with severe craniocerebral injury.Methods Forty-seven coma patients with severe craniocerebral injury were examined with BAEP and moaitored dynamically in early period of coma,and all these patients got head CT scans and the ratio of anteroposterior diameter and transverse diameter of mesencephalon were measured at the same time.Results There was a significantly negative relationship between the first BAEP classification and GOS score at the end of the follow-up (r =-0.755,P =0.000 ).Among all of the patients,there were 27 patients with the ratio of anteroposterior diameter and tansverse diameter of mesencephalon from 0.9 to 1.1,19 patients (70.4%,19/27) with good prognosis;20 patients with the ratio >1.1 or < 0.9,6 patients (30.0%,6/20 ) with good prognosis.The rate of good prognosis between the two had significant difference ( x2 =7.521,P =0.006).The sensitivity and specificity were 73.7%(14/19),82.6% (19/23) at the first BAEP combined with mesencephalic morphology,88.2% (15/17),100.0% (21/21) at the second BAEP combined with mesencephalic morphology.Conclusion BAEP combined with mesencephalic morphology can evaluate effectively the prognosis of coma patients with severe craniocerebral injury.
8.Efficacy analvsis of early hyperbaric oxygen therapy in patients with severe traumatic brain injury after craniotomy
Kangfeng LIU ; Wenjuan HUANG ; Fu HUANG ; Renduan CAI ; Zhihu YU ; Lei WANG ; Hua XIAO
Chinese Journal of Postgraduates of Medicine 2011;34(29):5-8
Objective To evaluate the effect of hyperbaric oxygen therapy (HBOT) in patients with severe traumatic brain injury (STBI) after craniotomy,and evaluate the difference of HBOT effects on the patients whose Glasgow coma score (GCS) were 6-8 scores and 3-5 scores.Methods Using case-control study,61 neurosurgical in-patients with STBl from February 6,2009 to November 25,2010 were divided into two groups by random digits table,30 in HBOT group and 31 in control group.Their GCS at the periods on admission,before HBOT and when they finished HBOT were recorded,as well as the Glasgow outcome score (GOS) before HBOT,finished HBOT,and 3 months after admission (GOS3M).Results The GCS finished HBOT were (12.63 ±2.70) scores in HBOT group and (11.64 ±2.50) scores in GCS 3-5 subgroup,there were statistically differences than those in control group [ (10.61± 3.01 ),(8.44 ± 1.67)scores] (P <0.05).The mean rank of GCS finished HBOT improvement (△GOSf) and GOS scores 3months after admission ( △ GOS3M) in HBOT group was 35.37 and 35.87,which were significantly higher than those in control group (26.77 and 26.29) (P <0.05).Meanwhile,the mean rank of △GOSf and △ GOS3M in GCS 3-5 subgroup was 12.14 and 13.09,which were significantly higher than those in control group (8.05 and 7.33) (P < 0.05 ).In GCS 6-8 subgroup,there was no significant difference in △ GOSf and △ GOS3M between HBOT group and control group (P > 0.05).Conclusion Early HBOT is effective to improve the recovery of consciousness and prognosis of the postoperative patient with STBI,especially of the patients with the special STBI (GCS 3-5 scores ).
9.New suspicious gene loci correlated with the generation and development of children 's retinoblastoma
Xiaoli ZHANG ; Weiling FU ; Jiangghua WANG ; Laixin ZHOU ; Manping LEI ; Chuan HUA
Chinese Journal of Tissue Engineering Research 2005;9(11):197-199
BACKGROUND: Recent researches have indicated that the generation and development of retinoblastoma(RB) might also be related with other anti-oncogenes except the known Rb1 gene.OBJECTIVE: To explore the loci of other genes which possibly participated in RB generation and development and try to find and confirm the indicators for the loss of heterozygosity(LOH) with merits in surveillance and prognosis.DESIGN: A case analysis by employing RB patients as subjects SETTING: A center of gene diagnosis and therapy of a military medical university-affiliated hospitalPARTICIPANTS: The study was conducted in the center of gene diagnosis and therapy of Xinan Hospital affiliated to Third Military Medical University of Chinese PLA. Sixteen RB cases including 10 males and 6 females were the patients of the outpatient department of three-affiliated hospitals of the Third Military Medical University of Chinese PLA from May 1998 to October 2001.Inclusion criteria: in accordance with RB diagnostic criteria and younger than 3 years old; Exclusion criteria: family heredity history. Two eyes were involved in 12 cases and one eye was involved in 4 cases.METHODS: Fourteen micro-satellite DNA labels on the 13th chromosome in tumor or peripheral blood samples were separately amplified by polymerase chain reaction(PCR) to analyze the incidence of LOH of each locus. Simultaneously, the genetic source of loci loss was confirmed by genealogical analysis.MAIN OUTCOME MEASURES: Frequency of LOH incidence on the 13th chromosome.RESULTS: In 16 RB patients,LOH occurred in one or more than one locus on the 13th chromosome of 12 cases. Thereinto, the probability of LOH occurrence on three loci including D13S265,D13S263 and D13S153(in Rb1gene) was the highest. Ten loci of LOH in 12 LOH positive samples were confirmed from agnate chromosomes. The RB confirmation of LOH-positive group or LOH-negative group needed 504 days or 1086 days,which was significantly different(t=2. 357,P<0.05).CONCLUSION: RB confirmation was earlier in LOH-positive patients than LOH-negative patients. Except the confirmed Rb1 gene, LOH on two loci including D13S263(13q14.1-14.2) and D13S265 (13q31-32) also might have certain suggestive effect on early intervention and functional surveillance of RB patients.
10.Sequence Analysis of Segment 8 of Five Chinese Isolates of Rice Gall Dwarf Virus and Expression of a Main Outer Capsid Protein in Escherichia coli
Ming-rong, DENG ; Xiao-lei, RUAN ; Fu-xiu, LIU ; Qin, ZHAO ; Hua-ping, LI
Virologica Sinica 2007;22(4):294-300
The rice gall dwarf disease, caused by the Rice gall dwarf virus (RGDV) is a serious disease occurring in rice in many regions of Guangdong province. As a basis to control the disease we have studied the genomic diversity of a variety of isolates from different locations. Genome segment 8(S8), encoding a main outer capsid protein (Pns8) of RGDV five isolates (BL, CH, DQ, GZ, XY) from Guangdong province was cloned and sequenced. The results revealed that all the S8 segments of the five isolates consisted of 1 578 nucleotides and had a single open reading frame (ORF) extending for 1 301 nucleotides from nucleotide 21 which encoded a polypeptide of 426 amino acids with an estimated molecular weight of 47.4 kDa. The S8 full-length sequence and the ORF sequence shared 97.3%-98.8% and 97.3%-99.1% nucleotide sequence identities within the five Chinese isolates, and shared 94.8%-95.6% and 95.0%-96.0% identities with those of the Thailand isolate respectively. The deduced amino acid sequence of Pns8 in GZ isolate was identical to that in the Thailand isolate, while the amino acid sequence variability of Pns8 within five Chinese isolates ranged from 0.5% to 2.1%. These results indicate that the S8 segment of RGDV is highly conserved in different isolates from different locations. The S8 cDNA from the XY isolate was cloned into the plasmid vector pET-28b(+) and a fused expression protein with an apparent molecular mass of 51kDa was specifically detected in an analysis of Escherichia coli Rossetta(DE3)Ⅱcells. To our knowledge, this is the first report on analysis of the RGDV segment 8 sequence and genetic comparison of different RGDV isolates and their protein expression.