1.Protective effects of ursodeoxycholic acid on α-naphthylisothi-induced acute liver injury in rats
Shibo LI ; Fangming XU ; Chuan XUE ; Xianjun DING ; Yuncheng LI ; Liyong QIAN ; Guoliang ZHANG ; Fang ZENG
Chinese Journal of Digestion 2012;32(5):325-329
ObjectiveTo investigate the protective effects and mechanism of ursodeoxycholic acid (UDCA) on α-naphthylisothi (ANIT)-induced cholestatic liver injury in rats.MethodsA total of 48 Sprague-Dawley (SD) rats were selected.Fouty-two rats were gavaged with ANIT (100 mg/kg) to induce acute liver injury,six rats were sacrificed 24 hours after the liver injury and the rats left were evenly divided into control group which were gavaged with saline and UDCA group which were gavaged with UDCA (20 mg/kg).Six rats were sacrificed at 48 hours,72 hours and 96 hours after modeling.The six untreated rats were set as blank control group.Serum and liver tissues of all rats were kept after sacrificed.Serum levels of alanine transaminase (ALT),aspartate transaminase (AST),total bilirubin (TBil) and total bile acid (TBA) were tested,interleukin-10 (IL-10),interleukin-6 (IL-6),and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA).The expression of multidrug resistance associated protein2 (Mrp2) at mRNA level in liver tissue was measured by quantitative real-time polymerase chain reaction (qRT-PCR) and the inflammatory reaction activity of liver tissues was inspected with Haematoidin-Eosin (HE)staining under microscope.ResultsAt 48 hours after liver injury modeling,serum TBil (143.80± 12.08) μmol/L vs.(178.50±15.19) μmol/L,TBA (13.15±3.81) μmol/L vs.(21.68±7.93)mol/L,IL-10 (44.13±3.68,37.15±6.25 ng/L),IL-6(50.80±2.09,57.32±4.63 ng/L) and TNF-α (17.53±0.84) ng/L vs,(19.10±1.64) ng/L of UDCA group and control group were compared,and the differences were statistically significant (P < 0.01 or P< 0.05).At 72 hours after liver injury modeling,serum ALT (721.67±97.54) U/L vs.(929.50±148.29) U/L and IL-10 (54.68±6.79)ng/L vs.(43.85±4.08) ng/L of UDCA group and control group were compared,and the differences were statistically significant (P<0.01 or P<0.05).At 96 hours after liver injury modeling,serum ALT (156.83±14.99) U/L vs.(250.67±42.29) U/L,AST (143.67±27.45) U/L vs.(206.00±63.94) U/L and TBil (23.53±5.08) μmol/L vs.(34.02±9.98) μmol/L of UDCA group and control group were compared,and the differences were statistically significant (P<0.01 or P<0.05).The differences of Mrp2 expression at mRNA level in liver tissues between UDCA group and control group at 48 hours (0.77 ± 0.21,0.46 ± 0.25),72 hours (2.27 ±0.84,1.10 ±0.38) and 96 hours (3.64±0.54,2.75±0.69) after liver injury modeling were statistically significant (P<0.01 or P<0.05).ConclusionThe mechanism of the protective effects of UDCA on ANIT-induced liver injury may be related with the regulation of serum cytokines and liver Mrp2 expression.
2.Use of gastrojugular shunt as minimally invasive portal systemic shunt before liver transplantation
Feng GAO ; Qian ZENG ; Xinchao YANG ; Fangming LIN ; Danni HUANG ; Hailong XUE
Chinese Journal of Hepatobiliary Surgery 2015;21(6):415-417
Surgical shunt is still an effective method in managing portal hypertension related gastrointestinal bleeding.To minimize the invasive trauma and adverse effect on transplantation remains to be the unsolved problem.Herein we present the use of a newly designed surgical shunt to cure massive refractory gastrointestinal tract hemorrhage in a patient,who was critically ill because of the extensive thrombus in portal venous system.The procedure is named gastrojugular shunt.For the sake of its simple operation and effective outcome,the procedure was performed on four other patients.All the patients were well treated and recovered uneventfully with good follow up results.
3.Purification of large-scale plasmid DNAs by selective precipitation with cetyltrimethylammonium bromide.
Quan ZHANG ; Kexiang YU ; Weifeng YUAN ; Fangming XUE ; Huaichang SUN ; Hongfei ZHU
Chinese Journal of Biotechnology 2008;24(12):2117-2121
Following Escherichia coli lysis with alkali, cetyltrimethylammonium bromide (CTAB) was directly titrated into the supernatant. An easy and feasible technology for plasmid purification was established with the optimized proportion between the quantity of CTAB and plasmid, combined with the specific solution for DNA release and TritonX-114 for endotoxin removal. Quality detection showed that the purified plasmid was free of contamination of host RNA. The host bacterial genomic DNA, endotoxin and bacterial protein were less than 10 microg, 50 EU and 10 microg per mg plasmids, respectively. The ratio of OD260/OD280 was between 1.75-1.85. Eighty percent of the prepared plasmids were presented in the supercoiled form. The plasmid purified with this technology can satisfy all criteria stipulated by FDA. The main advantages of the technology include the avoidance of animal-derived enzymes such as ribonucleases A, Proteinase K and toxic reagents like chloroform and phenol. In addition, the technology has low cost and no pollution.
Cetrimonium Compounds
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chemistry
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Chemical Precipitation
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DNA
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genetics
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isolation & purification
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DNA, Bacterial
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genetics
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isolation & purification
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Escherichia coli
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chemistry
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genetics
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Plasmids
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genetics
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isolation & purification
4.Analysis of the necessity of extraperitonealization in ileal conduit surgery after radical cystectomy
Fangming LIU ; Di JIN ; Wenhao SHEN ; Ming CAO ; Lianhua ZHANG ; Ruiyun ZHANG ; Liang DONG ; Wei XUE ; Yiran HUANG ; Haige CHEN
Chinese Journal of Urology 2017;38(5):352-356
Objective To discuss the necessity of closing the peritoneum during the operation of ileal conduit after the radical cystectomy.Methods We retrospectively analyzed the clinical data of 395 patients with bladder cancer who received radical cystectomy from Jan.2014 to Sep.2016.The amount of male was 327,female was 68.The mean age was (65.8 ± 9.7) years old.Patients were divided into four groups according to the surgical method of cystectomy and urinary diversion.In group A,patients,including 78 males and 9 females,were received open radical cysectomy (ORC) with extraperitoneal ileal conduit.The mean age was (67.8 ± 9.2) years old.In the preoperative clinical staging,66 cases were less than T2 and 21 cases were more than T2.Preoperative pathological grade in 83 cases and low grade in 4 cases.In group B,patients,including 31 males and 2 females,were accepted ORC with ileal conduit without peritoneum closure.The mean age was (67.3 ± 8.7) years old.Preoperative clinical staging showed less than T2 in 25 cases,more than T2 in 8 cases,The preoperative pathological grade showed high grade in 33 cases.In group C,patients,including 112 males and 27 females,were accpeted LRC with ileal conduit without peritoneum closure.The mean age was (64.3 ± 10.5)years old.The preoperative clinical staging showed less than T2 in 107 cases and more than T2 in 32 cases.The preoperative pathological grade showed high grade in 135 cases and low grade in 4 cases.In group D,patients,including 106 males and 30 females,were accepted RARC with ileal conduit without peritoneum closure.The mean age was (65.9 ±10.0)years old.Preoperative clinical staging showed less than T2 in 103 cases and more than T2 in 33 cases.The preoperative pathological grade showed high grade in 132 cases and low grade in 4 cases.Ileal conduit without peritoneum closure means completely open the peritoneum after anastomosis of the ureter and intestine in the urinary diversion surgery without shutting down the peritoneum,which is different from the extraperitoneal ileal conduit.The operating time,blood loss,blood transfusion rate,recovery time of intestinal function and perioperative complications and rate of hydronephrosis were analyzed.Results The 395 cases completed operation successfully,no LRC or RARC had been converted to ORC.The operative time was (280.1 ± 92.3) min,(233.6 ± 99.4) min,(304.8 ± 108.9) min,(364.6 ± 86.4) min in four groups,respectively (P < 0.05).The blood loss in four groups were (489.1 ± 285.6) ml,(431.8 ± 233.1) ml,(373.0 ±213.7) ml,(205.6 ± 137.8) ml,respectively (P <0.05).The transfusion rate in four groups were 18 (20.7%),16 (48.0%),15 (10.8%),14 (10.3%),respectively (P < 0.05).The mean time to flatus in four groups were (3.7 ±1.8)d,(3.6±1.0)d,(3.5±1.2)d,(2.2±1.7)d,respectively (P < 0.05).While ileal obstruction rate had no statistical difference in four groups [group A 17 cases(19.5%),group B 6 cases(18.2%),group C 27 cases(19.4%),group D 19 cases(14.0%),P =0.678].Urine leakage,intestinal leakage,lymphocyst were only occurred in group A [7 cases (8.0%),2 cases (2.3%),2 cases (2.3%)].Pyelonephritis was noticed in each group,including 14 cases(16.1%)in group A,2 cases(6.1%)in group B,9 cases (6.5%)in group C,6 cases(4.4%)in group D (P < 0.05).Hydronephrosis 6 months after surgery was observed in four groups,including 15 cases(17.2%)in group A,3 cases(9.1%)in group B,7 cases(5.0%)in group C,5 cases(3.7%)in group D (P < 0.05).Conclusions Ileal conduit without peritoneum closure would not increase the incidence of complications,on the contrary,it would relieve the tension of anastomosis,and reduce the occurrence of complications such as urine leakage.