1.Effects of recombinant human augmenter of liver regeneration on renal inflammation after renal ischemia reperfusion injury in rats
Xiaohui LIAO ; Hang SUN ; Qi LIU ; Hui GUO ; Ling ZHANG
Chinese Journal of Nephrology 2012;28(2):121-126
Objective To investigate the effects of recombinant human augmenter of liver regeneration (rhALR) on renal inflammation in acute kidney injury (AKI) induced by renal ischemia reperfusion (IR). Methods SD rats were randomly divided into sham-operated group,IR group,rhALR1 group (100 μg/kg) and rhALR2 group (200 μg/kg).Both renal pedicles of rats were identified and occluded with microvascular clamps for 60 min to induce acute kidney injury (AKI).Blood urea nitrogen and serum creatinine levels were evaluated using a Hitachi 747 automatic analyzer. For histological examination, sections were stained with HE. The activity of myeloperoxidase (MPO) was detected by spectrophotometer.Expression of TNF-α,ICAM-1,MCP-1 was determined by Western blotting. Results Blood urea nitrogen,serum creatinine levels and the injury of kidney were improved significantly in rhALR group as compared with IR group (all P< 0.05).They were improved more significantly in rhALR2 group as compared to in rhALR1 group (all P<0.05).The protein levels of TNF-α,ICAM-1,MCP-1 and the activity of MPO in kidneys from the sham-operated rats were low,and increased significantly after renal ischemia reperfusion injury (all P<0.05).After treated with rhALR,the expression of TNF-α,ICAM-1,MCP-1 and the activity of MPO were decreased significantly in kidneys as compared to those in IR group (all P<0.05),which decreased more significantly in rhALR2 group than those in rhALR1 group (all P< 0.05). Conclusions nhALR can protect kidneys from ischemia reperfusion injury in rats.The mechanism may be associated with the inhibition of renal inflammatory cells infiltration and down-regulated expressions of YNF-α,ICAM-1 and MCP-1 in the kidney.
2.Implication of expression of Nanog in prostate cancer cells and their stem cells.
Chen, GONG ; Hui, LIAO ; Fengjin GUO ; Liang, QIN ; Jun, QI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(2):242-6
Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells. The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells, and the relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells. By using magnetic active cell sorting (MACS), we isolated a population of CD44(+)/CD133(+) prostate cancer cells that display stem cell characteristics from PC3 cell line. Immunohistochemistry revealed positive expressions of CD44, CD133 and α(2)β(1)-integin in the isolated cells. CCK-8 analysis showed that isolated cells had a strong proliferative ability. The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing, indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line. Western blotting exhibited that the isolated cells had higher experession of Nanog, an embryonic stem marker, as compared with PC3 cells. Our study showed that Nanog might be helpful in sustaining the self-renewal and the undifferentiation of prostate cancer stem cells, and may serve as a marker for prostate cancer stem cells for isolation and identification.
3.Establishment of pTet-on rat precartilaginons stem cell strains and construction of pTRE-PTHrP(1-36) responsive plasmids
Shuwei ZHANG ; Anmin CHEN ; Minghui LI ; Jun QI ; Dengxin SONG ; Wentao ZHU ; Hui LIAO ; Fengjing GUO
Chinese Journal of Trauma 2008;24(7):564-568
Objective To establish a doxycycline-controlled immortalized pre-cartilaginons stem cells (IPCSCs) strains, clone parathroid hormone-related peptide[PTHrP(1-36)] gene and construct re- sponsive plasmid, pTRE-PTHrP (1-36). Methods Plasmid pTet-on was transfected into IPCSCs by using LipoinfectaminTM 2000 and then the stable clones were obtained by G418 screening. The doxycyc- line was added into the medium of monoclonal cells that were transiently transfected with plasmid pTRE- 2Hyg-Lue. The total RNA was extracted from PCSCs and the PTHrP(1-36) gene obtained by RT-PCR method. Then, the PTHrP (1-36) gene was subcloned to plasmids of Tet-responsive element with the se- lection marker of hygromycin pTRE-2Hyg to construct recombinant eukaryotic expression plasmid pTRE- PTHrP(1-36). After transferred into E. coli-DH5α, the clone was amplified, the recombinant plasm0ids were purified and identified by double-enzyme digestion. Results The doxycycline induced IPCSCs line was obtained, with 50 times higher than the non-induced cell line. Double enzyme digestion analysis and sequencing showed that the target gene was cloned into recombinant plasmid. Conclusions The induced IPCSCs line can be used to highly express alien genes. The responsive plasmid containing PTHrP (1-36) gene may be premising for rigorous control of PTHrp (1-36) gene expression.
4.Radiographic Measurement of Femoral Lateral Bowing and Distal Femoral Condyle Resection Thickness: Variances and Effects on Total Knee Arthroplasty Planning
Wu PEI?HUI ; Zhang ZHI?QI ; Gu MING?HUI ; Zhao XIAO?YI ; Kang YAN ; Liao WEI?MING ; Fu MING
Chinese Medical Journal 2017;(21):2557-2562
Background:Accurate evaluation of the plain radiography of lower limb is critical for preoperative planning of total knee arthroplasty (TKA). We aimed to investigate the effect of femoral lateral bowing and rotation on the radiographic measurements of distal femoral condyle resection thickness (DRT) and the distal femoral resection valgus angle (FVA). Methods: We analyzed 246 three?dimensional femoral models generated from computed tomography images of 123 patients, acquiring projected contours in seven positions – 20° and 10° internal rotation; 0° rotation; 10°, 20°, 30°, and 40° external rotation – for each model. Medial and lateral condyle DRTs, femoral shaft lateral bowing angle (FBA), and distal FVA were determined for each position. Linear mixed effect model was used to determine the effect of degree of femur rotation on repeated measurements of DRT or FVA. Results: FBA significantly affected the FVA and DRT (Pearson's R = 0.767 and ?0.408, respectively; P < 0.000). Samples were divided into three groups according to the FBA measured in neutral position: FBA <0°: DRT 3.75 ± 1.30 mm, FVA 4.53° ± 1.27°; FBA >0° but <3°: DRT 3.39 ± 1.31 mm, FVA 5.92° ± 1.31°; FBA >3°: DRT 2.22 ± 1.31 mm, FVA 7.37° ± 1.31°. From simulated 20° internal rotation to 40° external rotation in each femoral model, the average variation ranges of radiographically measured DRT, FVA, and FBA were 0.50 ± 0.28 mm, 2.93° ± 0.96°, and 10.33° ± 1.90°, respectively, with no significant differences among the FBA groups. The degree of femoral rotation significantly affected the FVA (F = 62.148, P < 0.000), whereas there was no effect on condyle resection thickness (F = 0.4705, P = 0.494). Conclusions: Axial femoral rotation has less effect on radiographic measurements of differences in the DRT than on those of the distal FVA.
5.Surgical treatment for both-column acetabular fractures using pre-operative virtual simulation and three-dimensional printing techniques
Huang JI-HUI ; Liao HUI ; Tan XIN-YU ; Xing WEI-RONG ; Zhou QI ; Zheng YU-SHI ; Cao HONG-YU ; Zeng CAN-JUN
Chinese Medical Journal 2020;133(4):395-401
Background:Surgical treatment of both-column acetabular fractures is challenging because of the complex acetabular fracture patterns and the curved surface of the acetabulum.Seldom study has compared the application of three-dimensional (3D) printing technology and traditional methods of contouring plates intra-operatively for the surgical treatment of both-column acetabular fractures.We presented the use of both 3D printing technology and a virtual simulation in pre-operative planning for both-column acetabular fractures.We hypothesized that 3D printing technology will assist orthopedic surgeons in shortening the surgical time and improving the clinical outcomes.Methods:Forty patients with both-column acetabular fractures were recruited in the randomized prospective case-control study from September 2013 to September 2017 for this prospective study (No.ChiCTR1900028230).We allocated the patients to two groups using block randomization (3D printing group,n =20;conventional method group,n =20).For the 3D printing group,1:1 scaled pelvic models were created using 3D printing,and the plates were pre-contoured according to the pelvic models.The plates for the conventional method group were contoured during the operation without 3D printed pelvic models.The operation time,instrumentation time,time of intra-operative fluoroscopy,blood loss,number of times the approach was performed,blood transfusion,post-operative fracture reduction quality,hip joint function,and complications were recorded and compared between the two groups.Results:The operation and instrumentation times in the 3D printing group were significantly shorter (130.8 ± 29.2 min,t =-7.5,P < 0.001 and 32.1 ± 9.5 min,t =-6.5,P < 0.001,respectively) than those in the conventional method group.The amount of blood loss and blood transfusion in the 3D printing group were significandy lower (500 [400,800] mL,Mann-Whitney U=74.5,P < 0.001 and 0 [0,400] mL,Mann-Whitney U =59.5,P < 0.001,respectively) than those in the conventional method group.The number of the approach performed in the 3D printing group was significantly smaller than that in the conventional method group (pararectus + Kocher-Langenbeck [K-L] approach rate:35% vs.85%;X2 =10.4,P < 0.05).The time of intra-operative fluoroscopy in the 3D printing group was significantly shorter than that in the conventional method group (4.2 ± 1.8 vs.7.7 ± 2.6 s;t =-5.0,P < 0.001).The post-operative fracture reduction quality in the 3D printing group was significantly better than that in the conventional method group (good reduction rate:80% vs.30%;X2 =10.1,P < 0.05).The hip joint function (based on the Harris score 1 year after the operation) in the 3D printing group was significantly better than that in the conventional method group (excellengood rate:75% vs.30%;x2 =8.1,P < 0.05).The complication was similar in both groups (5.0 % vs.25 %;x2=3.1,P =0.182).Conclusions:The use of a pre-operative virtual simulation and 3D printing technology is a more effective method for treating bothcolumn acetabular fractures.This method can shorten the operation and instrumentation times,reduce Mood loss,blood transfusion and the time of intra-operative fluoroscopy,and improve the post-operative fracture reduction quality.
6.Effects of lead on thyroid function of occupationally exposed workers.
Qi-rong LIANG ; Rui-qin LIAO ; Su-hua SU ; Shu-hai HUANG ; Rui-hui PAN ; Jia-le HUANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2003;21(2):111-113
OBJECTIVETo explore the effects of lead on the thyroid function of occupationally exposed workers.
METHOD157 workers occupationally exposed to lead in a smelting factory were investigated. The concentration of lead in air at workshop was measured by flame atomic absorption spectrophotometry (FAAS) and the levels of blood lead (PbB) by atomic absorption spectrophotometry (AAS), zinc protoporphyrin (ZPP) by ZnPP meter, as well as the indexes of thyroid function, thyroid-stimulating hormone (TSH), triiodothyronine (T(3)), thyroxin (T(4)), free T(3) (FT(3)), and free T(4) (FT(4)) in serum by radioimmunoassay.
RESULTSThe workers with higher level of blood lead (> 2.88 micro mol/L) showed lower levels of T(3) [(1.54 +/- 0.39) nmol/L] and FT(3) [(5.50 +/- 1.26) pmol/L] than those with lower blood lead level [PbB: (1.92 approximately 2.88) micro mol/L group, T(3): (1.71 +/- 0.45) nmol/L, FT(3): (6.12 +/- 1.64) pmol/L, P < 0.05]. There was no obvious effect of length of service on thyroid hormone of exposed workers.
CONCLUSIONHigher level of blood lead may cause certain damage to thyroid function by inhibiting deiodination of T(4). No obvious relation between length of service and thyroid function was found.
Adult ; Female ; Humans ; Lead ; blood ; toxicity ; Male ; Middle Aged ; Occupational Exposure ; Thyroid Gland ; drug effects ; physiology
7.Cloning, Sequence Analysis and Expression of dhaT Gene from Citrobacter freundii and Purification and Property of Corresponding Recombinant Enzyme
Xiang-Hui QI ; Zhao-Fei LUO ; Yu-Tuo WEI ; Fa-Zhong CHEN ; Shan-Shan WANG ; Shou-Hai HOU ; Dong-Qing LIAO ; Ri-Bo HUANG ;
China Biotechnology 2006;0(07):-
1,3-propanediol (1,3-PD) is an important material for chemical industry, therefore, there is much interest in the production of 1,3-PD. The gene dhaT encoding 1, 3-propanediol dehydrogenase ( PDOR) of Citrobacter freundii was amplified by PCR. Sequence analysis of the similarity at the nucleotide and amino acid level between the gene encoding C. freundii PDOR and that of C. freundii ( U09771 ) were 78% and 90% , respectively. The recombinant plasmid pSE-dhaT was constructed by inserting dhaT gene into expression vector pSE380 and then transformed E. coli JM109. The recombinant strain was induced by IPTG to express dhaT. Further more the recombinant enzyme was purifed from recombinant E. coli by Ni-nitrilotriacetate affinity chromatography followed by Sephacral S-300 gel filtration. A single obvious protein about 42kDa could be obtained by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis of recombinant enzyme. The purified enzyme was used to determined enzyme property on the substrate of propionaldehyde and 1, 3-PD. The optimal temperature and optimal pH of the purified enzyme were 37℃, 8.0 for reduction and 25℃, 10. 5 for oxidation, respectively; and the kinetic property of PDOR about Km and V max were 10. 05mmol/L, 37. 27?mol/min/mg for propionaldehyde and 1. 28mmol/L, 25. 55?mol/min/mg for 1,3-PD, respectively; The deduced dhaT gene product (388 amino acids) showed a specific reduction activity of 49. 50U/mg and oxidation activity of 79. 92U/ mg. There also have a putative iron-binding motif ( G-XX-H-X-X-A-H-X-X-G-X-X-X-X-X-P-H-G) as a fingerprint pattern in the recombinant enzyme, the motif is fully conserved among these 1, 3-propanediol dehydrogenase. It is beneficial to the researches of high producing 1, 3-propanediol by gene engineering strain.
8.Phylogenetic Analysis of Homologous Proteins Encoded by UL2 and UL23 genes of Herpesviridae
Long-ding, LIU ; Wen-juan, WU ; Min, HONG ; Hai-jing, SHI ; Shao-hui, MA ; Jing-jing, WANG ; Hong-ling, ZHAO ; Yun, LIAO ; Qi-han, LI
Virologica Sinica 2007;22(3):207-211
The proteins encoded by the Herpesviridae β-gene play a critical role in the replication stage of the virus. In this paper, phylogenetic analyses provided evidence that someβ-gene products, such as UL2 and UL23 from HSV1, have their homologous genes in its family, and also exist in prokaryotic organisms, indicating that these viruses appear to have been assembled over evolutionary time by numerous independent events of horizontal gene transfer.
9.Implication of Expression of Nanog in Prostate Cancer Cells and Their Stem Cells
GONG CHEN ; LIAO HUI ; GUO FENGJIN ; QIN LIANG ; QI JUN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(2):242-246
Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells.The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells,and fhe relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells.By using magnetic active cell sorting (MACS),we isolated a population of CD44+/CD133+ prostate cancer cells that display stem cell characteristics from PC3 cell line.lmmunohistochemistry revealed positive expressions of CD44,C D 133 and α2β1-integin in the isolated cells.CCK-8 analysis showed that isolated cells had a strong proliferafive ability.The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing,indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line.Western blotting exhibited that the isolated cells had higher experession of Nanog,an embryonic stem marker,as compared with PC3 cells.Our study showed that Nanog might be helpful in sustaining the self-renewal and fhe undifferentiation of prostate cancer stem cells,and may serve as a marker for prostate cancer stem cells for isolation and identification.
10.Methyltransferase inhibitor BIX01294 promotes the migration and inhibits decidualization of mouse uterine stromal cells in vitro.
Hui-Qi LIAO ; Liu TIAN ; Hui YANG ; Ni MA ; Chang-Jun ZHANG ; Hong-Lu DIAO
Journal of Southern Medical University 2017;37(6):730-736
OBJECTIVETo investigate the effect of BIX01294 (BIX), a methyltransferase inhibitor, on the migration and decidualization of the stromal cells in mouse uterus.
METHODSMouse endometrial stromal cells were isolated and cultured from the uterus of pregnant mice on day 3.5 of gestation. The migration and decidualization of mouse endometrial stromal cells treated with BIX at different concentrations were observed with wound healing assay and real-time PCR.
RESULTSThe migration distance of mouse endometrial stromal cells increased as the BIX concentration increased within the range below 15 µmol/L. Compared with the control cells, the cells treated with BIX (15 µmol/L) showed significantly increased migration distances, but increasing BIX concentration to 20 µmol/L did not further increase the cell migration distance and began to cause cell death. Compared with the control cells, the BIX-treated stromal cells exhibited significantly down-regulated expression of Ehmt2 mRNA, and 15 µmol/L BIX caused inhibition of decidualization in the stromal cells.
CONCLUSIONWithin a defined concentration range, BIX promotes the migration and inhibits decidualization of mouse uterine stromal cells by inhibiting the expression of Ehmt2 mRNA.