1.Prokaryotic Expression and Purification of a Cotton Dehydration Responsive Element Binding Protein GhDBP1 and Its DNA Binding Activity
Progress in Biochemistry and Biophysics 2006;33(3):247-253
Cotton (Gossypium hirsutum) is one of the most important economic crops in the world. Its growth and productivity were affected by environment stresses such as drought, cold and high salinity. Thus, the enhanced stress tolerance in this plant is of great importance. As the dehydration responsive element (DRE) binding protein (DBP) plays an important role in the regulation of plant resistance to environmental stresses and is quite useful for generating transgenic plants tolerant to these stresses, isolation and functional analysis of DBPs in cotton are important to cotton production. In the previous work, a DBP gene from cotton, named as GhDBP1, was isolated and its expression patterns in cotton plants was demonstrated at the transcriptional level. Here, the expression,purification and DNA binding activity of GhDBP1 were reported. The entire coding region of the GhDBP1 gene was inserted into an expression vector, pET28a, and transformed into Escherichia coli BL21 (DE3). The fusion protein was successfully expressed under IPTG induction and the purified recombinant protein was obtained by Ni-NTA affinity chromatography. Non-radioactive electrophoretic mobility shift assay revealed that the purified GhDBP1 protein was able to form a specific complex with the previously characterized DRE element. In addition, the computer modeling of the DNA-binding domain of GhDBP1 were performed using SWISS-MODEL software. The main-chain structures and the folding patterns of the DNA-binding domain of GhDBP1 were similar to the known structure of the DNA-binding domain of the Arabidopsis thaliana GCC box-binding protein AtERF1. These results indicate that GhDBP1 is a DRE-binding transcription factor and might use the structure similar to that of AtERF1 to interact with DRE sequence.
2.Progress in diagnosis and treatment of anastomotic leakage after anterior resection of rectal cancer
International Journal of Surgery 2014;41(1):46-51
Anastomotic leakage was one of the severe complications after anterior resection of rectal cancer.Its occurrenc was correlated with age,weight and nutrtion,basic diseases,preoperative radio-chemotherapy,surgical technique and stage of the tumor.Anastomotic leakage could be diagnosed by symptoms,physical signs,drainagefluid and imaging examinations.Most of the anastomotic leakage could be cured non-operatively.Much attention should be pay on the prevention of anastomotic leakage,protective stomy should be made in high risk population,while the complications of protective stomy should be pay attention to.
3.PLCE1 modulates p53 expression and apoptosis in esophageal cancer cells
Yun LI ; Junhang ZHANG ; Jun AN ; Jinyuan HE ; Shaohong HUANG
Chinese Pharmacological Bulletin 2015;(1):82-86
Aim To investigate the role of phospho-lipase C epsilon 1 ( PLCE1 ) in modulating the apoptot-ic mechanism in esophageal cancer ( Eca ) cells. Methods Eca cell lines, OE33 and CP-C cells were cultured to assess the expression of PLCE1 . siRNA suppress expression of PLCE1 . The expression of PLCE1 and p53 was evaluated by quantitative real time PCR and Western blot. Methylation analyses of p53 were performed by bisulfite conversion of genomic DNA. Apoptosis was assessed by flow cytometry. Results OE33 and CP-C cells expressed high levels of PLCE1 . Knockdown of PLCE1 markedly increased the expression and hypomethylation of p53 , and in-creased the frequency of apoptosis. Conclusion PLCE1 suppresses apoptosis of Eca cells via promoting p53 promoter methylation and inhibiting expression of p53 .
4.Preoperative evaluation of local infiltration of rectal cancer using high-resolution 3.0T MR
Zheng WANG ; Zhongkui HUANG ; Jinyuan LIAO ; Ningqin LI ; Rui SONG
Journal of Practical Radiology 2017;33(8):1196-1200
Objective To investigate the value of high-resolution 3.0T MR in the assessment of local infiltration of preoperative rectal cancer.Methods A total of 168 patients pathologically proved rectal cancer underwent both conventional pelvic and rectal high-resolution before operation, and the imaging findings were reviewed retrospectively.The accuracy of preoperative high-resolution 3.0T MR in prediction of pathological staging was assessed,and the characteristic imaging features of local infiltration in preoperative rectal cancer were discussed.Results The relationship between circumference invasion of colorectal cancer and the pathological T staging was moderately positive (rs=0.530,P=0.003).Compared the staging of colorectal cancer on MRI with pathologic T staging,the overall diagnostic accuracy was 84.52%,and there was a stronger correlation between MRI findings and pathological staging (rs=0.837,P=0.001).The best single parameters for diagnosing T3 stage rectal cancer on MRI were nodular convex of the tumor and muscular signal interruption,with 91.1% specificity and 89.7% sensitively respectively.And the best combination of parameters was the cord appearence of intestinal wall and muscular signal interruption,with 89.3% specificity and 78.0% sensitively respectively.Conclusion High-resolution 3.0T MR can be preferable to evaluating local infiltration of rectal cancer, showing a higher clinical value to asseee T staging of preoperative rectal cancer.
5.Determination of diosgenin and ruscogenin in Radix Ophiopogonis by nonaqueous capillary electrophoresis.
Baomei HUANG ; Chengwei YAO ; Qingquan BIAN ; Zhiguo WANG ; Jinyuan MO
Acta Pharmaceutica Sinica 2011;46(4):443-6
Nonaqueous capillary electrophoresis is used for the determination of the contents of diosgenin and ruscogenin in Radix Ophiopogonis. The operating buffer was composed of 20 mmol x L(-1) Na2B4O7-HCl (pH 7.61) in 70% methanol. The applied voltage was 25 kV and detection potential was at +0.70 V. With these conditions, the components were successfully separated. The content of diosgenin in Radix Ophiopogonis was 0.018 mg x g(-1) and ruscogenin was 0.008 mg x g(-1). The average recoveries of diosgenin and ruscogenin were 102% and 99.2%, respectively. A new method of the quality control of diosgenin and ruscogenin in Radix Ophiopogonis is provided.
6.The value of multi-slice spiral CT liver perfusion imaging to evaluate the chronic hepatic fibrosis and cirrhosis
Liling LONG ; Zhongkui HUANG ; Ke DING ; Jinyuan LIAO ; Jianning JLANG
Chinese Journal of Radiology 2012;46(4):317-321
ObjectiveTo investigate the value of the MSCT liver perfusion imaging parameters inthe evaluation of the chronic hepatic fibrosis and cirrhosis. Methods Liver CT perfusion ( CTP ) was performed in 107 participants,including 31 patients with mild hepatic fibrosis( S1,S2),34 patients with severe hepatic fibrosis ( S3,S4 ) and early stage of hepatic cirrhosis which conformed by liver pathologic biopsy,42 patients with hepatic cirrhosis who had typical clinical and image signs,and 30 healthy subjects as control group.The data of CTP ( HAP,PVP,LTP,HPI and TTP) at different stages were obtained with Body perfect CT-syngo CT2007A and control study with histopathologic stage.Compared the study index by the one-way ANOVA analysis. Used Spearman rank correlation to analysis the relationship between liver perfusion imaging parameters and the degrees of the chronic hepatic fibrosis. Used Logistic regression to analysis the maximum.regression coefficient among the liver perfusion imaging paraneters,which affected the histopathologic stage mostly.ResultsIn the subgroups of the chronic hepatic fibrosis S1,S2,S3,S4 to the hepatic cirrhosis,HAP values was (28.9 ±8.6),(24.6 ±2.4),(29.2 ±2.3) and (38.9 ± 7.0) ml · 100 ml -1 · min-1,respectively.HAP decreased firstly,then increased.Statistic analysis showed the difference of HAP between later-stage cirrhosis and other groups( F =40.26,P < 0.01 ).PVP values of above subgroups was (111.3 ± 18.1),(92.9 ±5.3),(73.0 ±9.0) and (54.1 ± 13.8) ml · 100 ml-1 ·min -1,respectively.TLP values of above subgroups was ( 140.2 ± 25.9 ),( 117.1 ± 4.5 ),( 102.3 ± 8.7 )and (93.0 ± 11.8) ml · 100 ml-1.min-1,respectively.The difference of PVP,TL.P among each subgroup was significant ( F =136.79,67.40,respectively,P < 0.01 ).HPI values of above subgroups was (20.4 ± 2.6)%,(21.0 ±2.1)%,(28.5 ±3.1)% and (42.6± 11.1)%,respectively.TTP values of above subgroups was (123.7±22.2),(137.1 ±27.1),(145.0 ±28.6) and (166.5 ±25.1)s,respectively.The difference of HPI,TTP among each subgroup was significant( F =93.05,17.37,respectively; P <0.01 ).PVP,TLP was significant negative correlation with the degree of the hepatic fibrosis( r =-0.920,-0.846,respectively; P <0.01 ).HAP,HPI and TTP was significant positive correlation with the degree of the hepatic fibrosis( r =0.611,0.882 and 0.545,respectively; P < 0.01 ).Logistic regression analysis showed the regression coefficient of PVP( - 8.798) was maximum.With an area under the receiver operating characteristic curve of PVP =84.76 ml · 100 ml- 1 · min- 1 as a diagnose critical point.The sensitivity was 0.890,the specificity was 0.950,and the accuracy was 0.931 in the prediction of the chronic hepatic fibrosis.Conclusions MSCT liver perfusion imaging parameters can reflect the hemodynamic changes of chronic hepatic fibrosis and cirrhosis.CTP may be helpful for differentiation the severe hepatic fibrosis and early stage of hepatic cirrhosis and later-stage cirrhosis.
7.Treatment of children spleenomegaly by partial splenic embolization
Fan LIU ; Zenghui QIN ; Liangbo XU ; Sui HUANG ; Jinyuan YANG
Journal of Interventional Radiology 1994;0(04):-
Objective To investigate the safety treatment of partially embolizing spleenomegaly in children. Methods Forty two children aged 1-15 with spleenomegaly were treated through staged partial splenic embolization (PSE). The first embolized scope of spleen was 30%-40% with the second being 30%-40% at 1 or 2 months later in order to achieve the goal of getting rid of hypersplenia and improving the splenic function. Results The adverse effects of splenic embolisation were slight with short duration of fever and stomachache and with efficient control of hypersplenia and its correlative basic diseases except one case of splenic abscess.Conclusions Spleenomegaly in Children can be more safely and more efficiently cured through staged PSE.
8.Effects of N-acetylcysteine on Alcoholism in Mice
Yongping HUANG ; Shiwen ZHOU ; Ying XU ; Jianlin TANG ; Lian WANG ; Jinyuan ZHANG ; Xue YANG
China Pharmacy 2001;0(07):-
OBJECTIVE:To investigate the effects of N-acetylcysteine(NAC)on the concentration of ethanol in serum and the activities of alcohol dehydrogenase in liver and stomach tissue in alcoholic mice.METHODS:All mice were given p.o white spirit30min after p.o saline or5%NAC solution,and the durations from the disappear of righting reflex to recovery in mice were recorded and the number of dead mice in24hours were countered;With the same procedure for six days,the mice were killed by withdrawaling blood from orbit,and taken out the liver and stomach immediately,the concentrations of ethanol in serum and the activities of alcohol dehydrogenase in liver and stomach were measured by chromatometry and colorimetric method respectively.RESULTS:In drunk test,5%NAC solution significantly postponed the time from p.o white spirit to the disappear of righting reflex(P
9.Significance of the detections for CD4 +CD25 + regulatory T cells, Foxp3 mRNA and interleukin 2 receptor in kidney transplantation recipients
Jun TIAN ; Jinyuan ZHANG ; Nanmei LIU ; Dayong HU ; Weifeng HU ; Jian HUANG
Chinese Journal of Nephrology 2008;24(7):466-470
Objective To observe the changes of CD4+CD25+ regulatory T cells, Foxp3 mRNA and soluble interlukin 2 receptor (sIL-2R) in the peripheral blood of kidney transplantation recipients and to evaluate their effect on the diagnosis of acute rejection. Methods Forty-two renal transplant recipients and 30 healthy controls were enrolled in this study. CD4+CD25+ regulatory T cells proportion, Foxp3 mRNA and sIL-2R of pre-transplantation and those of day 7,14, 28, 56 of post-transplantation were measured by flow cytometer, fluorescent quantization PCR and enzyme-linked immunosorbent assay (ELISA), respectively. Biochemistry appliance was used to detect serum creatinine. The diagnosis of acute rejection in transplanted kidney was based on the clinical symptoms, the laboratory examinations, Doppler ultrasound and biopsy. Results (1)At day 7, 14, 28, 56 of post-transplantation, CD4+CD25+ regulatory T ceils proportion, Foxp3 mRNA level in acute rejection group were significantly decreased compared with those in non-acute rejection group. (2) There were significant differences of peripheral blood CD4+CD25+ regulatory Tcells[(9.22±3.53)% vs (6.09±1.99)%, P<0.01], Foxp3 mRNA[(0.82±0.36)×10-3 vs (0.50±0.28)×10-3, P<0.01] and sIL-2R levels [(856.30±108,24) U/ml vs (247.35±11.24) U/ml, P<0.01]between patients of pre-transplantation and healthy control group. (3)Plasma CD4+CD25+ regulatory T cells [(16.53±4.14)%] and the expression of Foxp3 mRNA [(4.97±1.94)×10-3] was significantly increased, but sIL-2R level [(463.72±31.23) U/ml] was significantly decreased as the transplanted renal function was restored (all P<0.01). (4) Plasma CD4+CD25+regulatory T cells [(12.18~2.86)%] and the expression of Foxp3 mRNA [(3.15±1.22)×10-3] was significantly decreased (P<0.01), and sIL-2R level [(748.36±115.41) U/ml] was significantly increased (P<0.01) when acute rejection occurred. The above changes had an earlier onset than the change of Scr. (5)The percentage of CD4+CD25+ regulatory T cells was positively correlated with the Foxp3 mRNA level (P<0.01), but was not correlated with sIL-2R level in all the patients. Conclusion The measurement of these markers in peripheral blood may be an important guideline to the diagnosis and prognosis of acute rejection in renal transplant recipients.
10.PLCE1 suppresses apoptosis of lung adenocarcinoma cells via modulating p53 expression
Yun LI ; Xueping LUO ; Jizong LIN ; Shaohong HUANG ; Jun AN ; Jinyuan HE ; Junhang ZHANG
Chinese Journal of Pathophysiology 2015;(1):119-123
AIM:To investigate the role of phospholipase C epsilon 1 ( PLCE1 ) in modulating the apoptotic mechanism in lung adenocarcinoma A 549 cells.METHODS:PLCE1 inhibitor U-73122 was used to suppress the expres-sion of PLCE1.The expression of PLCE1 and p53 in A549 cells was evaluated by quantitative real-time PCR and Western blotting.Apoptosis was assessed by flow cytometry .RESULTS:A549 cells expressed high level of PLCE1 and low level of p53.Inhibition of PLCE1 markedly increased the expression of p 53, and increased the apoptosis of A 549 cells.CON-CLUSION:PLCE1 suppresses apoptosis of A549 cells via inhibiting the expression of p53.