2.Effect of Shen warming Pi strengthening method on the expression of serum T cell subsets in IBS-D rats.
Xiao-lan SU ; Ru-han WEI ; Wei WEI ; Jing ZHANG ; Yi-bing BAI ; Hai-xia SHI
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(4):457-460
OBJECTIVETo observe the effect of Shen warming Pi strengthening method on expressions of serum T cell subsets (C045+%, C03+%, and C04 +/COB+) in diarrhea-predominant irritable bowel syndrome (IBS-0) rats. Methods An IBS-0 rat model was established referring to AL-Chaer's modeling method combined with tail clamp and intragastric administration of sanna leaf. After modeling 30 SO rats were randomly divided into 6 groups according to random digit table, i.e., the model group, the high, middle, low dose Wenshen Jianpi Recipe (WJR) groups, and the Sishen Pill control group, 6 in each group. A normal control group consisting of 6 SO rats were also set up. Rats in high, middle, low dose WJ R groups were administered by gastrogavage with boil-free WJ R at the daily dose of 3. 100, 1. 550, 0. 775 g/kg, respectively. Rats in the Sis hen Pill control group were administered by gastrogavage with boil-free Sis hen Pill at the daily dose of 0. 736 g/kg. Equal volume of normal saline was given by gastrogavage to rats in the model group and the normal control group. All medication lasted for 2 successive weeks. Rats' general state, expressions of T cell subsets (CD45+%, CD3+%, and CD4+ /CDB+) changes were observed.
RESULTSCompared with the normal control group, expressions of CD45+% and CD3+% increased, but CD4+ /CDB+ decreased with statistical difference (P < 0. 05). Compared with the model group, expressions of CD45+% and CD3+% decreased, but CD4+ ICDB+ increased with statistical difference in high, middle, low dose WJR groups, and the Sis hen Pill control group (P <0. 05). Compared with the Sis hen Pill control group, there was statistical difference in all indices except CD45+ value in the low dose SWPSM group (P <0. 05). Compared with the low dose WJ R group, the expression of CD3+% decreased in high and middle dose WJR groups, and the Sis hen Pill control group; CD4+ /CD8+ increased in the Sishen Pill control group and the high dose SWPSM group (all P < 0. 05).
CONCLUSIONSWJR showed better treatment effect. The mechanism of Shen warming Pi strengthening method might be achieved by regulating expressions of CD45+% and CD3+%, and CD4+ /CD8+ ratios.
Animals ; Drugs, Chinese Herbal ; Female ; Irritable Bowel Syndrome ; therapy ; Leukocyte Common Antigens ; metabolism ; Medicine, Chinese Traditional ; Rats ; T-Lymphocyte Subsets ; metabolism
3.Cloning and High Expression Anabaena Class-II Fructose-1, 6-bisphosphate Aldolase Gene in Escherichia coli
Lan-Zhen WEI ; Yi-Wen CUI ; Wei-Min MA ; Quan-Xi WANG ;
China Biotechnology 2006;0(05):-
The complete genomes of more cyanobacterial strains have been completed for sequence, and genetic engineering of cyanobacteria has evolved in the post-genome era. Since Kaneko and colleagues had completed the sequence for the complete genome of Anabaena sp. PCC 7120 in 2001, functions of some genes in this genome, including fructose-1,6-bisphosphate aldolase (FBA) gene, have been predicated using the method of bioinformatics. However, little information is available regarding whether this gene can encode FBA and its product characteristics of related enzyme. Here, to explore this information, the predicted II-FBA gene-encoding region in Cyanobase database was cloned by PCR method and then ligated into pET-32a to generate the expression vector, pET-FBA-II. The results of SDS-PAGE indicated that the expression level of the expected target polypeptide was approximate 23.4 percent as compared to total protein and the molecular weight is about 40 kDa as compared to the protein molecular marker. The results of enzyme activity analysis showed that the activity of II-FBA was ~11.8 U per mg protein and owned a standard activity of II-FBA. To sum up, the results not only prove the functional prediction of this II-FBA gene from the Cyanobase database, but also provide the important conditions for further studying its physiological and biochemical characteristics and functions of the gene expression product.
5.Pirfenidone effects on human hypertrophic scar fibroblasts cultured in vitro
Wei LAN ; Xiaojian LI ; Xueliang JI ; Xianfeng YI ; Yanzhi LIU ; Rongmei TU
Chinese Journal of Tissue Engineering Research 2015;(24):3808-3812
BACKGROUND:Studies have shown that cytokine inhibitor pirfenidone can inhibit biological activity of fibroblasts by regulating a variety of cytokines. It has made good progress in the research and application of anti-fibrosis of internal organs, but the effect and mechanism for hypertrophic scars and skin fibroblasts are unclear. OBJECTIVE:To investigate the effect of pirfenidone on human hypertrophic scar fibroblasts. METHODS:Human hypertrophic scar fibroblasts were cultured using tissue culture method. Passages 3-6 cel s grew wel in the logarithmic growth phase were col ected. Cel s were divided into the control group (0 g/L pirfenidone), 0.15, 0.3 and 1 g/L pirfenidone groups according to different mass concentrations. Cel s were intervened for 12, 36 and 48 hours. RESULTS AND CONCLUSION:MTT, reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay results demonstrated that compared with the control group, cel proliferation, transforming growth factorβ1 mRNA expression, types I and III col agen secretion were decreased in the 0.15, 0.3 and 1 g/L pirfenidone groups (P<0.05), and the decrease was most significant in the 1 g/L pirfenidone group (P<0.05). At 24, 48 and 72 hours after intervention, significant differences in inhibitory rate of cel proliferation and the secretion of types I and III col agen were detected among 0.15, 0.3 and 1 g/L pirfenidone groups (P<0.05). Results confirmed that pirfenidone apparently inhibited the secretion of col agen of hypertrophic scar fibroblasts cultured in vitro, transforming growth factorβ1 expression and cel proliferation and viability.
6.Expression of SET8 in renal cell carcinoma tissue and its clinical significance
Xiuwang WEI ; Xiaoming YI ; Chaopeng TANG ; Zhenyu XU ; Jianbo LIANG ; Zhixiang LAN ; Wenquan ZHOU
Journal of Medical Postgraduates 2015;28(10):1048-1052
Objective The occurrence and progression of renal cell carcinoma ( RCC) is complicated process associated with DNA abnormal methylation , histone modification , and Wnt signaling pathway .This study aimed to investigate the expression of histone methylase SET8 in RCC, its relationship with the Wnt signaling pathway , its action mechanism in RCC , and its clinical significance . Methods We selected 50 cases of RCC treated by radical nephrectomy , detected the expression of SET 8 in the RCC and adjacent noncancerous kidney tissues by immunohistochemical EliVision two-step staining with β-catenin.We compared the expression levels of SET8 and β-catenin in the two types of tissue and analyzed their relationship with the patients′clinical information and the pathologic stage and grade of tumor as well as the correlation between the SET 8 andβ-catenin expressions . Results SET8 was mainly express in the cytoplasm of the RCC and noncancerous kidney tissues , partially in the cell membrane and nucleus , while theβ-catenin protein chiefly in the cell membrane of renal tubular epithelial cells in the normal kidney tissue .The expression levels of SET 8 and β-catenin in the RCC tissue were closely related to the TNM stage and tumor grade (P<0.05).The positive expression of SET8 in the RCC tissue (76%[38/50]) showed no significant difference from that in the ad-jacent noncancerous kidney tissue (66% [33/50]) (P>0.05), but that of β-catenin was remarkably higher in the former (68%[34/50]) than in the latter (4%[2/50]) (P<0.01).There was a positive correlation between the positive expression of SET 8 and the abnormal expression of β-catenin (r=0.219, P<0.05). Conclusion SET8-activated H4K20me-1 controls the activation and abnormal activities of the Wnt signaling pathway , affects the gene transcription and cell activity , and participates in the occurrence , progression, and distant metastasis of RCC .
7.Identification of up-regulated miRNAs in extrahepatic and intrahepatic cholangiocarcinoma
Changzheng LIU ; Wei LIU ; Jingjing LI ; Yi ZHENG ; Lan YU ; Xiaodong HE ; Songsen CHEN
Chinese Journal of Hepatobiliary Surgery 2012;18(6):466-469
Objective To investigate the expression profile of miRNAs up-regualted in human extrahepatic and intrahepatic cholangiocarcinoma tissues and probe the effect on cell growth of four of these miRNAs in QBC939 cell line.Methods Up-regulated miRNAs in extrahepatic or intrahepatic cholangiocarcinoma tissues were analyzed by using miRNA-microarray,which was confirmed by using miRNA Real-Time PCR analysis.Based on these findings,four of these up-regulated miNRAs were chosen to perform function investigation.The specific miRNA inhibitors were transfected into QBC939 cells,respectively,and cell proliferation assay was performed by using MTT.Results 12 miRNAs were up-regulated both in two types of cholangiocarcinoma tissues,28 miRNAs and 21 miRNAs were up-regulated in extrahepatic cholangiocarcinoma and intrahepatic cholangiocarcinoma,respectively.MiR-125b and miR-19a expression levels were increased about 3.7 and 3.6 fold,compared with the matched normal bile duct tissues (P<0.05).MiR-92a and miR-205 expression was upregulated about 4.S- and 3.5-fold,compared with the matched normal bile duct tissues (P<0.05).MiR-125b,miR19a,miR-21,and miR 378* were inhibited in QBC939 cells,which indicated a significant inhibitory effect on cell growth.The ratio of inhibition was 71%,72%,69%,and 76%(P<0.05)at 36 h,61%,63%,60%,and 59%(P<0.01) at 48 h,and 61%、56%、60% and 59%(P<0.05) at 60 h.Conclusion The miRNAs expression patterns in human extrahepatic and intrahepatic cholangiocarcinoma tissues are different and uo-regulated miRNAs act as oncomirs on cholangiocarcinoma cell growth.
8.Effect of alcohol extract from Coreopsis tinctoria Nutt. on spontaneous hypertension rats by metabolomic methods
Wei LU ; Xiaoli MA ; Yi LAN ; Linlin LI ; Ye WANG ; Chunli JIN ; Xinmin MAO
Chinese Pharmacological Bulletin 2014;(9):1311-1314,1315
Aim To investigate the effects of the alco-hol extract from Coreopsis tinctoria Nutt. on urinary metabolomics of spontaneous hypertension rats ( SHR) . Methods SHR were fed with normal diet for 1 week and then, they were randomly divided into six groups:untreated control, the high, middle, low dos-age group of the alcohol extract(3.2 g·kg-1 ,1.6 g· kg-1,0.8 g·kg-1) , captopril group(4 mg·kg-1) and Duzhong tablet group(187.5 mg·kg-1). The u-rine of normal rats and SHR hypertension model rats was collected on 1,2,3,4 weeks. The metabolic pro-files were analyzed using 1 H-NMR. PLS-DA methods were used to discriminate the difference and the bio-markers. Results Compared with model group, the blood pressure of all groups was significantly lowered after 4 weeks ( P <0.01 ) . The alcohol extract from Coreopsis tinctoria Nutt. could significantIy reduce blood pressure, and the urinary metabolic profiles trea-ted with Coreopsis tinctoria Nutt. were changed signifi-cantly such as IIL,creatine,β-glucose,etc. Conclusion The alcohol extract from Coreopsis tinctoria Nutt. could significantIy reduce blood pressure and change the urinary metabolomics profiles of SHR rats.
9.Evaluation of nephron-sparing surgery for renal cell carcinoma
Qun WAN ; Zhou-Jun SHEN ; Zhong-Yi LI ; Wei-Feng LAN ; Yang-Cheng MAO ;
Chinese Journal of Urology 2001;0(07):-
Objective To evaluate the indications and intraoperative management of nephron-spar- ing surgery (NSS) for renal cell carcinoma (RCC).Methods A total of 53 patients (32 men and 21 women;age range,21 -76 years) with unilateral RCC and a normal contralateral kidney underwent NSS.The disease course ranged from 1 week to 13 months.The unilateral RCCs were detected by B-uhrasonography. Partial nephrectomy was performed on 38 cases of polar RCC and wedge nephrectomy was performed on 15 cases of central RCC.Both renal vein and artery were blocked for 10 - 15 min,and unclamped for 1 -2 min if necessary.The resections were finished within 7 -21 min (mean,12 min).No inosine and regional hypo- thermia were used during operation.Absorbable hemostatic gauze and terylene flake were used to suture renal wound surface.After operation all patients were treated with interferon hypodermic injection of 5 000 000 U every other day for 3 months.Results The diagnosis of RCC with negative cutting margin was confirmed by pathological examination in all patients.The greatest dimension of the renal cancers were<4 cm except for one of 6 cm.The clinical stage was T_1N_0M_0 in all patients.During the follow-up of 6 -48 months,all pa- tients were alive without recurrence and had normal renal function.Conclusions NSS can be rapidly per- formed for T_1 stage renal cancer<4 cm or single clearly localized cancer>4 cm in diameter.The follow-up results are satisfactory.
10.Apply research of MRI combined with 1H-MRS in meningiomas
Wei LI ; Wan-sheng LONG ; Xue-mao LUO ; Xiao-qin CAI ; Yong LAN ; Yi-gai HE ;
Journal of International Oncology 2010;37(11):877-880
Objective To evaluate the roles of magnetic resonance imaging and proton magnetic resonance spectroscopy(1H-MRS) in the diagnosis of meningiomas. Methods 98 patients with meningiomas underwent conventional pre-contrast MR and contrast MR. Among them, 28 cases had two dimensional single voxel or multi voxel 1 H-MRS simultaneously both in the lesion's region and the contralateral side. Results On precontrast MR images of 98 cases, T1 WI showed 58.1% (61/105) isointensities, 31.4% (33/105) faintly low intensities and 10. 5% (11/105) mixed intensities; T2WI showed 40. 0% (42/105) isointensities, 41.0%(43/105) hyperintensities, 10.5% (11/105) faintly low intensities and 8.5% (9/105) mixed intensities. After administration of Gd-DTPA, the solid part of the tumors exhibited various enhancement in all the 98 cases.28 cases of MRS exhibited specific different spectral peaks, including increased of choline-containing compounds(Cho), absent or decreased of acetylaspartate(NAA), and the unchanged of creatine(Cr). The value of NAA, Cr, Cho, NAA/Cr, Cho/Cr, NAA/Cho in the tumor center of meningioma were 0. 09 ± 0.06,0.31 ± 0. 22, 0.46 ± 0. 16, 0.33 ± 0. 42, 1.50 ± 0. 68, 0. 15 ± 0.08, compared with the contralateral normal region, Cr has no significant difference (P > 0. 05), NAA, Cho, NAA/Cr, Cho/Cr, NAA/Cho had significantly differences(P < 0.05). Conclusion Conventional pre-contrast MR and contrast MR is the most important dignostic means for meningiomas, 1H-MRS combined with MRI can improve the diagnostic accuracy of meningiomas.