1.Experimental study of ectopic activation about allograft segment tibia of rabbits
Yanping LUO ; Mingwu ZHOU ; Ruifu YANG ; Yang LI ; Libo XU ; Xun ZHANG
Chinese Journal of Microsurgery 2015;38(1):61-65
Objective To compare of the difference about ectopic activation between autogenous bone graft and allograft from large segment tibia of rabbits.Methods Eighty healthy adult Chinese rabbits (6 months of age),weighing (2.5 ±-0.5)kg,were randomly divided into experimental group (allogeneic bone group) and the control group (autograft group),40 rabbits in each group.Another 10 rabbits were allogeneic bone donor.In experimental group,when 1.5 cm long rabbit tibial allograft were finishied,they were implanted into spatium intermusculare between the musculus rectus femoris and medial vastus muscle of the rabbit around the saphenous artery and were fastened to the femur by 1.0 mm Kirschner-wire.In control group,autologous tibias were done,the same as experimental group including length and position and method.Four weeks and 8 weeks and 12 weeks postoperative,respectively,the postmortem specimens were examined gross and immunohistochemistry and the expression of BMP-2 and collagen type Ⅰ of transplanted bone tissue were detected.Results BMP-2 mainly exist in cytoplasm of osteoblasts and chondrocytes undifferentiated mesenchymal cells.Collagen type Ⅰ primarily exist in the bone matrix around the pit of bone.The expression level of BMP-2 of experimental group in postoperative 4,8,12 and 16 weeks were 85.25 ± 4.47,109.44 ± 14.69,141.85 ± 9.45,116.25 ± 14.18,respectively,and the expression level of BMP-2 of control group were 103.78 ±-6.59,124.95 ± 14.94,145.46 ± 8.10,112.48 ± 13.27,respectively.The expression level of collagen type Ⅰ of experimental group in postoperative 4,8,12 and 16 weeks were 78.74 ± 7.99,95.95 ± 6.99,139.91 ± 4.32,137.76 ± 3.48,respectively,and the expression level of BMP-2 of control group were 88.87 ± 11.26,102.45 ± 2.82,140.76 ± 4.62,139.05 ± 4.55.Compared with control group,there was a significant difference in the expression level of the BMP-2 and collagen type Ⅰ of experimental group in postoperative 4,8 weeks (P < 0.05),but,there was no significant difference in the expression level of the BMP-2 and collagen type Ⅰ of experimental group in postoperative 12,16 weeks (P > 0.05).The amount increased gradually during 4 weeks,8 weeks,12weeks,peaked at 12 weeks,BMP-2 displayed a downward trend at 16 weeks,and collagen type Ⅰ basiclly maintain the level of 12 weeks.Conclusion Allograft segment could complete activation while they are implanted into spatium intermusculare containing famous blood supply within 3 months,there is no significant difference between autologous bone and allograft,it shows the feasibility of ectopic activation about allograft segment.
2.Role of miR-129-5p in regulation of epithelial-mesenchymal transition of peritoneal mesothelial cells
Xun ZHOU ; Fuyou LIU ; Ying LUO ; Dan TANG ; Shikun YANG ; Lin SUN ; Li XIAO
Chinese Journal of Nephrology 2015;31(4):269-276
Objective To investigate the role of microRNA-129-5p (miR-129-5p) in the regulation of epithelial-mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs) isolated from peritoneal dialysate effluents and TGF-β1 induced HPMCs line.Methods The isolated cells were cultured from peritoneal dialysate effluents overnight of 10 patients just started PD and 12 patients with PD over 6 months.Taqman PCR assay was used to determine the expression of miR-129-5p in the HPMCs.Moreover,the expression of miR-129-5p in HPMCs induced by 5 μg/L TGF-β1 for 0-72 h was also detected by Taqman PCR.HPMCs were pre-transfected with miR-129-5p precursor (pre-mir-129-5p) to overexpress miR-129-5p,then incubated with TGF-β1 for 48 h,and the expression of EMT associated gene and protein was detected by real-time PCR,Western blotting and immunofluorescence,respectively.Furthermore,the effect of TGF-β1 on the expression of Smad interacting protein-1 (SIP1) and the regulation of pre-miR-129-5p on the SIP1 expression also were investigated.Results MiR-129-5p expression significantly down-regulated in the HPMCs isolated from PD patients over 6 months than from PD start patients(P < 0.01).Similarly,TGF-β1 remarkably decreased miR-129-5p in HPMCs lines on time-dependent manner (P < 0.01).Pre-mir-129-5p dramatically restored the expression of epithelial marker E-cadherin,while inhibited the expression of Vimentin,a mesenchymal marker,in HPMCs induced by TGF-β1 (all P < 0.01).In addition,TGF-β1 increased SIP1 expression in HPMCs time dependently,while the high level of SIP1 protein was obviously repressed after transfected of pre-miR-129-5p (P < 0.01),but there was no obvious change of its mRNA expression.Conclusion MiR-129-5p modulates EMT formation of HPMCs in PD process,possibly by posttranscriptional inhibition of SIP1.Targeting miR-129-5p/SIP1 may provide a new approach for the prevention and treatment of peritoneal fibrosis during PD.
3.Intraperitoneal application of chemotherapy in cervical cancer
Yaqi LI ; Xun GONG ; Lianying LUO ; Yan MA ; Yu ZHANG ; Weiting YANG ; Zhongxiu WU ; Guofang LIAO
Chinese Journal of Primary Medicine and Pharmacy 2012;19(2):203-204
Objective To observe the clinical efficacy and toxicity of intraperitoneal chemotherapy combined with intravenous chemotherapy in treatment of cervical cancer.Methods The experimental group of 40 patients received systemic chemotherapy of docetaxel and cisplatin combined abdominal intraperitoneal high-frequency hyperthermia in the control group,40 patients received docetaxel plus cisplatin chemotherapy,The efficacy and toxicity were observed.Results The complete remission rate of observotion group was 82.5%,significantly higher than 62.5% in control group.Effective rate in observation group was 92.5% and also significantly higher than 75.0% in control gronp.After 6 cycles of chemotherapy,the improvement of the experimental group was 85.0% higher than 57.5 % in control gronp.The differemles between the two groups was statistically significant(P < 0.05); the main side effects during chemotherapy are gastrointestinal reactions,since the cells decreased,hair loss,the difference was not statistically significant(P >0.05).Conclusion The chemotherapy of docetaxel and cisplatin intraperitoneal chemotherapy can significantly improve the treatment of cervical cancer,without increasing side effects.
4.Association of c.553G>T polymorphism in the apolipoprotein A5 gene with coronary heart disease and the levels of serum lipid
Fang QIU ; Kui ZHANG ; Lei LI ; Guang-Yu GU ; Li WANG ; Xun-Yang LUO ; Yong-Quan XIA ; Ping GU ;
Chinese Journal of Laboratory Medicine 2003;0(07):-
T site polymorphism is closely related to CHD and elevated serum triglyceride and total cholesterol.
5.Clinical implication of urinary protein markers in diabetic nephropathy and interventional effects of Chinese herbal medicine.
Xi-Miao SHI ; Xian-Jie MENG ; Yi-Gang WAN ; Shan-Mei SHEN ; Xun-Yang LUO ; Liu-Bao GU ; Jian YAO
China Journal of Chinese Materia Medica 2014;39(14):2589-2594
In clinic, some urinary protein makers can dynamically and noninvasively reflect the degree of renal tubular injury in patients with diabetic nephropathy (DN). These urinary biomarkers of tubular damage are broadly divided into two categories. One is newfound, including kidney injury molecule-1 (Kim-1), neutrophil getatinase-associated lipocalin (NGAL), liver-type fatty acid-binding protein (L-FABP) and cystatin C (CysC); the other one is classical, including beta2 microglobulin (beta2-MG), retinal binding protein (RBP) and N-acetyl-beta-D-glucosaminidase (NAG). It is reported that, the increases in urinary protein markers are not only closely related to the damage of tubular epithelial cells in DN patients, but also can be ameliorated by the treatment with Chinese herbal compound preparations or Chinese herbal medicine. Recently, although urinary proteomics are used in the protein separation and identification, the traditional associated detection of urinary protein markers is more practical in clinic. At present, it is possible that the associated detection of urinary biomarkers of glomerular and tubular damages may be a feasible measure to reveal the clinical significance of urinary protein markers in DN patients and the interventional effects of Chinese herbal medicine.
Biomarkers
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urine
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Diabetic Nephropathies
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complications
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drug therapy
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urine
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Drugs, Chinese Herbal
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pharmacology
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therapeutic use
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Humans
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Medicine, Chinese Traditional
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methods
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Proteinuria
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complications
6.Characteristic of urinary protein spectrum in patients with stage III diabetic nephropathy and its regression analysis with traditional Chinese medicine symptom.
Yi-Gang WAN ; Xian-Jie MENG ; Shan-Mei SHEN ; Xun-Yang LUO ; Liu-Bao GU ; Xi-Miao SHI ; Jian YAO
China Journal of Chinese Materia Medica 2013;38(23):4157-4163
To analyze the characteristic of urinary protein spectrum in patients with stage III diabetic nephropathy (DN) and its compliance with traditional Chinese medicine (TCM)symptom, for the sake of providing a basis for clarifying the rules of TCM syndrome differentiation in DN. Adopting the traditional epidemiological retrospective method, thirty-eight TCM syndromes and urinary protein with medium or low molecular weight, as well as urinary enzyme, including 24 h urinary protein (Upro), urinary albumin( UAlb), urinary retinal binding protein( URBP), urinary cystatin C (UCysC), urinary N-acetyl-beta-D-glucosaminidase (UNAG), were collected from 108 patients with stage III DN, and a multiple factor regression analysis between them was conducted. As the results, the levels of Upro, UAlb, URBP, UCysC, and UNAG were increased in 108 patients with stage III DN. Qi-Yin deficiency type was the major type. The level of UAlb in patients with Qi-Yin deficiency type was significantly higher than those without Qi-Yin deficiency type (P < 0.05). The elevation of Upro with the factors as swift digestion with rapid hungering, lassitude and lack of strength, weakness of waist and knees was complied, the elevation of UA1b with the factors as dry mouth with desire to drink, the elevation of URBP with the factors as numbness of extremities, shortness of breath, the elevation of UCysC with the factors as clear urine in large amounts, and the elevation of UNAG with the factors as frequent micturition, were complied respectively. In conclusion, for 108 stage III DN patients. The increase in urinary protein spectrum including UAlb, URBP, UCysC, and UNAG is the major characteristic. Shen and Pi are the major organs related to the appearance of urinary protein; Pi-Shen deficiency is the basic pathogenesis. The level of UAlb is taken as one of the objective syndrome factors for Qi-Yin deficiency type. The levels of UNAG and UCysC are possibly the objective syndrome factors for Shen-Qi deficiency type.
Diabetic Nephropathies
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complications
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diagnosis
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urine
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Female
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Humans
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Male
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Medicine, Chinese Traditional
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methods
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Middle Aged
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Proteinuria
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complications
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urine
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Qi
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Regression Analysis
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Yin-Yang
7.Dynamic variation of endothelin in portal hypertensive patients with hemorrhage and its clinical significance.
Hong-wu LUO ; Fei-zhou HUANG ; Xun-yang LIU ; Shu-ping REN
Journal of Central South University(Medical Sciences) 2005;30(4):427-429
OBJECTIVE:
To investigate variations of plasma endothelin (ET) and its clinical significance in portal hypertensive patients with esophageal variceal hemorrhage.
METHODS:
Sixty-six patients with portal hypertension were randomly divided into 2 groups. Group I (32 patients) received general therapy and Group II (34 patients) received general therapy and UTI after hemorrhage. The plasma ET concentration and liver function were determined at 1, 2, 4, 7, 10, and 14 d after the hemorrhage. Another 20 patients without the hemorrhage were elected as the control group.
RESULTS:
At 7 and 14 d after the hemorrhage, the levels of TBIL, ALT and AST were elevated at first and then decreased in Groups I and II. The decrease of TBIL, ALT and AST levels was significantly faster in Group II than in Group I (P < 0.05, P < 0.01, P < 0.05, respectively) on 14 d after the hemorrhage. At 1 d after the hemorrhage the ET concentration was markedly increased in Group I and II as compared with the control group (P < 0.01). Then it was gradually decreased on 10 d after the hemorrhage. The ET concentration in Group II was decreased more rapidly than that in Group I on 2, 4 and 7 d after the hemorrhage (P < 0.05; P < 0.01; P < 0.05, respectively). The ET concentration was positively correlated to TBIL levels in groups I and II (r = 0.734, P < 0.01). And the decreased index of ET concentration was negatively correlated to the increased index of TBIL (r = -0.486, P < 0.05).
CONCLUSION
The increased plasma ET in portal hypertensive patients with hemorrhage may contribute to liver injury. UTI can protect the liver function by inhibiting ALT, AST, TBIL and ET level.
Adult
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Aged
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Endothelin-1
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blood
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Esophageal and Gastric Varices
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blood
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etiology
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Female
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Glycoproteins
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therapeutic use
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Humans
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Hypertension, Portal
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blood
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complications
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Liver Failure
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prevention & control
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Male
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Middle Aged
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Trypsin Inhibitors
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therapeutic use
8.Akt gene therapy for cirrhotic rats with portal hypertension.
Fei-zhou HUANG ; Gang DENG ; Xun-yang LIU ; Cheng-qun LUO
Journal of Central South University(Medical Sciences) 2008;33(1):31-37
OBJECTIVE:
To determine whether there is an impaired Akt and eNOS activation in cirrhotic livers, and to investigate the feasibility of transferring adenovirus-mediated Akt gene to the liver for portal hypertension.
METHODS:
Recombinant adenovirus Ad-myr-HA-Akt and Ad-EGFP were produced by homologoas recombination in 293 cells . The Methods of compound factor, carbon tetrachloride (CCl4), corn flour, and cholesterol plus alcohol were used to construct the hepatic cirrhosis rat models. Ten normal rats were served as a normal control group, and 40 cirrhotic rats were divided into 4 groups randomly: an untreated group, an Ad-myr-HA-Akt treated group, an Ad-EGFP group, and a saline group. Ad-myr-HA-Akt, Ad-EGFP, and saline were transduced into the Ad-myr-HA-Akt treated group, Ad-EGFP group, and saline group via the tail vein respectively. Portal vein pressure, mean arterial pressure, and heart rate were measured in all rats. Protein abundance and phosphorylation status of Akt and eNOS were examined by Western blot. Spectrophotometry was used to measure the NO level. Frozen sections of the liver, heart, lung, kidney, brain, spleen, and testis were made to examine the expression of enhanced green fluorescent protein (EGFP) by fluorescence microscopy on Day 3 in the Ad-EGFP group.
RESULTS:
The concentration of recombinant adenovirus Ad-myr-HA-Akt after the purification was 5.5 x 10(11)vp/mL and that of Ad-EGFP was 6.0 x 10(11)vp/mL. Akt and eNOS phosphorylations in the liver of cirrhotic rats were obviously impaired. Adenoviral delivery of myr-Akt restored eNOS phosphorylation, increased the NO level and decreased the portal pressure after 3 days of adenoviral infection. In contrast, the livers infected with Ad-EGFP and saline were not changed. The EGFP expression was mainly found under the fluorescence microscopy on the frozen section of liver. Very little fluorescence was detected in the lung and kidney; and there was no detectable EGFP in other organs.
CONCLUSION
There is an impaired Akt and eNOS activation in the cirrhotic livers; myr-Akt gene therapy can restore the Akt activation and NO production in the cirrhotic liver, suggesting that this therapy may be helpful in treating portal hypertension.
Adenoviridae
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genetics
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metabolism
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Animals
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Carbon Tetrachloride
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Carbon Tetrachloride Poisoning
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Genetic Therapy
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Hypertension, Portal
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etiology
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therapy
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Liver Cirrhosis, Experimental
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complications
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metabolism
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therapy
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Male
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Nitric Oxide Synthase Type III
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metabolism
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Proto-Oncogene Proteins c-akt
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genetics
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Random Allocation
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Rats
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Rats, Sprague-Dawley
10.Dynamic accumulation analysis on bioactive constituents of Polygonum multiflorum in different collection periods.
Yi-yuan LUO ; Juan-xiu LIU ; Xun-hong LIU ; Cai-wu LAN ; Ya HOU ; Yang MA ; Sheng-nan WANG ; Bao-chang CAI
China Journal of Chinese Materia Medica 2015;40(13):2565-2570
To study the dynamic change law of bioactive constituents from Polygonum multiflorum, and to explore the optimal harvest period of P. multiflorum. Determination of stilhene glucoside, anthraquinones and catechin from P. multiflorum in different harvest times by MEKC-DAD, and principal component analysis (PCA) was used to comprehensive evaluation for bioactive constituents. There are obvious differences among the contents of active ingredients in various collecting periods samples, the content of stilbene glucoside was the highest in November, the total content of combined anthraquinone was the highest in November and December, the content of catechin was the highest in September. The comprehensive evaluation index obtained with principal component analysis showed that the sample collected in November is significantly higher than those with other samples. The optimal harvest period of P. multiflorum is November.
Electrophoresis
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Fallopia multiflora
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chemistry
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growth & development
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metabolism
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Time Factors