1.Preparation of polyclonal antibody for minoxidil, development of an ELISA method for detection of minoxidil
Biao WEI ; Guirong ZHANG ; Danshen ZHANG ; Lingyun NIE ; Yanping TANG
Chinese Journal of Immunology 2015;31(12):1668-1673
Objective:On the basis of synthetic antigens on minoxidil ,we prepared its polyclonal antibody and established a ELISA method to detect minoxidil .Methods:We synthesized minoxidil artificial antigen with glutaraldehyde and identified by UV scan -ning.We had established a detection of minoxidil competition ELISA .Results: The UV scan showed that the minoxidil successfully coupled to a carrier protein.The conjugate of minoxidil-BSA was used to immunize BALB/c mice.And the produced antiserum showed high titer of 1:12 800 in the indirect ELISA.Its ELISA curve equation:y=-0.082 3 x+0.938 ( R2 =0.9811 ) , minoxidil mass concentration and the absorbance in 0.001-10 μg/ml showed a good linearity .Conclusion: This method has been successfully preparing minoxidil artificial antigen and polyclonal antibodies and established ELISA detection method for minoxidil ,which provides a basis for the practical application of minoxidil immunoassay method .
2.AAV-HGFK1 inhibits proliferation of colorectal cancer cells by down-regulating the phosphorylation of EGFR
Feihong DENG ; Biao NIE ; Junhua ZUO ; Xuehua LIU ; Jinmin CHEN
Chongqing Medicine 2014;(33):4447-4449,4453
Objective To study the effect of adeno associated virus hepatocyte growth factor K1(AAV‐HGFK1)on the prolifer‐ation of 4 different colorectal cell lines with or without KRAS or BRAF mutation .Methods The levels of epidermal growth factor receptor (EGFR) mRNA were determined in SW48 without KRAS or BRAF mutation ,Lovo with KRAS mutation ,SW620 with KRAS mutation ,HT29 with BRAF mutation by quantitative real time PCR ,respectively .After the infection of AAV‐HGFK1 ,the expressions of EGFR ,p‐EGFR and β‐actin were detected by Western blot and the proliferation of the cells were assayed using MTT .Results Lovo ,SW48 and HT29 expressed EGFR protein while SW620 did not .EGF promoted the proliferation of Lovo , SW48 and HT29 cells .AAV‐HGFK1 down‐regulated the phosphorylation of EGFR and significantly inhibited their proliferation . But EGF had no effect on proliferation of SW620 stimulated by EGF .Conclusion AAV‐HGFK1 exhibited its antitumor effects through EGF/EGFR signaling irrespective of the KRAS or BRAF mutation and may also act through other signaling pathways .
3.Application of CT venography in diagnosis and treatment of Budd-Chiari syndrome
Xiaogao WANG ; Shiyuan CHEN ; Biao SHI ; Xiaochun QIAO ; Yong GAO ; Chaowen YU ; Zhonglin NIE
Chongqing Medicine 2016;45(8):1043-1045
Objective To investigate the application value of CT venography(CTV) in the diagnosis and treatment of Budd‐Chiari syndrome(BCS) .Methods 58 patients with BBCS in our hospital from January 2012 to January 2014 were performed the CTV examination .The inferior vena cava ,hepatic vein ,portal vein and collateral vessels were performed the reconstruction analysis . Results All the patients were definitely diagnosed as BCS after completing CTV examination ,including :19 cases of inferior vena cava(IVC) diaphragm type ,15 cases of short IVC segment occlusion ,8 cases of long IVC segment occlusion ,9 cases of hepatic vein stenosis or occlusion ,7 cases complicated by fresh thrombosis .In addition ,the different degrees of compensatory expansion of col‐lateral vesse ,intuition and comprehensiveness ,can display the position ,character and length of lesion vessel ,also observes the de‐grees of collateral vessels expansion and liver cirrhosis .
4. Effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells and ciprofloxacin on Staphylococcus aureus in vitro
Biao ZHOU ; Hualei TU ; Te BA ; Lingfeng WANG ; Shujie WANG ; Shunyi NIE
Chinese Journal of Burns 2017;33(6):355-360
Objective:
To explore the effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells (hUCMSCs) and ciprofloxacin on
5.Establishment and comparison of liver metastasis models of two colorectal carcinoma cell lines in mice.
Jun-bao WEN ; Biao NIE ; Bo JIANG
Journal of Southern Medical University 2007;27(7):1044-1046
OBJECTIVETo establish hepatic metastasis models of two colorectal carcinoma cell lines in mice for studying mechanism involved in colorectal carcinoma metastasis and its potential countermeasures.
METHODSMurine and human colorectal carcinoma CT26 and LoVo cells were inoculated into the spleen of Balb/c mice and Balb/c nude mice, respectively. The conditions of all the mice were observed, and the survival time and liver metastases were recorded.
RESULTSAll mice inoculated with CT26 cells and a few with LoVo cells developed liver metastases without metastases in any other organs. Pathological examination identified the liver metastatic foci as poorly differentiated colonic adenocarcinoma. Compared with the mice inoculated with LoVo cells, those with CT26 cells had a higher rate of liver metastasis and a shorter survival time.
CONCLUSIONThe mouse model has been established successfully, which well mimics the pathological process of liver metastasis of colorectal cancer.
Animals ; Cell Line, Tumor ; Colorectal Neoplasms ; pathology ; Disease Models, Animal ; Drug Evaluation, Preclinical ; Female ; Humans ; Liver ; pathology ; Liver Neoplasms ; drug therapy ; pathology ; secondary ; Mice ; Spleen ; pathology
6.p16 and MGMT gene methylation in sputum cells of uranium workers.
Shi-biao SU ; Lu-jing YANG ; Wei ZHANG ; Ya-li JIN ; Ji-hua NIE ; Jian TONG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(2):92-95
OBJECTIVETo study the methylation of O-6-methylguanine-DNA methyltransferase (MGMT) and p16 gene in the sputum cells of radon-exposed population. To provide the experimental base for finding the molecular biomarker of the high risk population of the radon-induced lung cancer.
METHODS91 radon-exposed workers were divided into 4 groups, high dosage group (> 120 WLM), middle dosage group (between 60 and 120 WLM), low dosage group (between 30 and 60 WLB) and lower dosage group (between 2 and 30 WLM) according to the accumulated exposure dosage of the radon daughters. The abnormal methylation of p16 and MGMT gene in the sputum cells of the population in the four groups was detected with the methylation specific PCR (MSP).
RESULTSThere was significantly upward trend for the p16 gene methylation rate (0.00%-20.00%), the MGMT gene methylation rate (0.00%-28.00%) and the total methylation rate (0.00%-40.00%) with the increase of the accumulated exposure dosage of the radon daughters (P < 0.01).
CONCLUSIONThe methylation of p16 and MGMT gene is related to the accumulate exposure dosage of the radon daughters.
Carcinogens, Environmental ; adverse effects ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; DNA Methylation ; Dose-Response Relationship, Radiation ; Humans ; Male ; O(6)-Methylguanine-DNA Methyltransferase ; genetics ; metabolism ; Occupational Exposure ; Radon ; adverse effects ; Radon Daughters ; adverse effects ; Sputum ; metabolism
7.nm23-H1 gene inhibits lung cancer cell invasion through down-regulation of PKC signal pathway.
Qiang NIE ; Qing-hua ZHOU ; Wen ZHU ; Lun-xu LIU ; Jun-ke FU ; Ding-biao LI ; Yin LI ; Guo-wei CHE
Chinese Journal of Oncology 2006;28(5):334-336
OBJECTIVETo study the molecular mechanisms of nm23-H1 for regulating PKC signal pathway before and after transfection with nm23-H1 gene.
METHODSUsing Western-blot, Boyden-chamber, MTT and laser scanning confocal microscopy (LSCM) techniques to detect the distribution of PKC in cytosol and plasma membrane, changes of invasion and proliferation activity, PKC translocation status and changes of intracellular Ca(2+) concentration among different human pulmonary carcinoma cells with transfected or untransfected nm23-H1 gene, and changes of the three cell lines after treatment with Calphostin C, a PKC inhibitor.
RESULTS(1) The expression of PKCalpha, PKCbeta II on L9981 and L9981-pLXSN cell membrane, which was in activated status, was remarkably higher than those in L9981-nm23-H1 cell line (P < 0.001). The expression of PKCalpha, PKCbeta II in cytosol in L9981 and L9981-pLXSN cell lines, which was in inactivated status, was lower than those in L9981-nm23-H1 cell line (P < 0.001). It means that the PKC signal pathway was activated in L9981 and L9981-pLXSN cell lines. (2) PKCalpha and PKCbeta II mainly located in nuclei and perinuclear area in L9981 and L9981-pLXSN cells, which were in active status, and the Ca(2+) concentration in these cells was obviously higher than that in L9981-nm23-H1 cell line (P < 0.01). In L9981-nm23-H1 cell line, which was transfected with nm23-H1 gene, PKCalpha and PKCbeta II mainly located in soluble cytosolic section, in an inactive status. (3) The invasion and proliferation ability of L9981 and L9981-pLXSN lung cancer cells was higher than that of L9981-nm23-H1 cell line (P < 0.001). There was no statistically significant difference between L9981 and L9981-pLXSN cell lines (P > 0.05). (4) After treated with PKC inhibitor Calphstin C, the expression of PKC and PKCbeta II in membrane in L9981 and L9981-pLXSN cell lines was down-regulated (P < 0.001), PKCalpha and PKCbeta II were mainly located in cytosolic area, mainly in an inactive status, and the Ca(2+) concentration was found to be decreased in all the three cell lines. The invasion and proliferation ability of the three lung cancer cell lines were obviously decreasing (P < 0.001). However, the invasion and proliferation ability of L9981-nm23-H1 lung cancer cell line was still lower than that of L9981 and L9981-pLXSN lung cancer cell lines (P < 0.001). There was also no significant difference between L9981 and L9981-pLXSN cell lines (P > 0.05).
CONCLUSIONThe results of this study suggest that nm23-H1 gene might inhibit the invasion and metastasis of lung cancer cells by down-regulating PKC signaling pathway. The Ca(2+) in cells might be involved in this process.
Calcium ; metabolism ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Cell Proliferation ; drug effects ; Cytosol ; metabolism ; Down-Regulation ; Humans ; Lung Neoplasms ; enzymology ; metabolism ; pathology ; NM23 Nucleoside Diphosphate Kinases ; genetics ; Naphthalenes ; pharmacology ; Neoplasm Invasiveness ; Protein Kinase C ; antagonists & inhibitors ; metabolism ; Protein Kinase C beta ; Protein Kinase C-alpha ; metabolism ; Signal Transduction ; Transfection
8.Comparison of detection of trisomy 8 with fluorescence in situ hybridization and conventional karyotype analysis in myelodysplastic syndrome.
Zhi-Biao ZHANG ; Shi-He LIU ; Juan LI ; Li-Jin BO ; Huan-Ying CUI ; Xu-Ping LIU ; Yan-Xia NIE ; Shuang QIN
Journal of Experimental Hematology 2002;10(2):115-118
The purpose of this study was to compare the detection of trisomy 8 in myelodysplastic syndrome (MDS) patients with interphase fluorescence in situ hybridization (FISH) and cytogenetic karyotype analysis. Using Spectrum Green labeled chromosome 8 centromere probe, interphase FISH was established. The trisomy 8 clones were simultaneously detected in 48 MDS cases with FISH and conventional cytogenetic analysis (CCA). Results showed that the CCA revealed no significant difference of constitutional proportion between MDS-RA and MDS-RAEB with karyotypes of whole +8, partial +8 and one +8. With FISH, detectable rates were 66.1% for whole +8. Partial +8 and sole +8 were significantly higher than one +8 and complex +8, respectively. The percentages of trisomy 8 were similar in MDS-RA and MDS-RAEB. Trisomy 8 was detected in 1 of 15 specimens with normal or abnormal karyotype without trisomy 8 by FISH. There was linear correlation between the percentages of partial +8 detected by FISH and CCA. Two patients received CCA and FISH examination at diagnosis and during treatment, the percentage of trisomy 8 was increased with progress of disease. In conclusion, our results showed that FISH is a sensitive and accurate technique to detect trisomy 8 in MDS patients. It can provide contribute to diagnosis, assessment of curative effect and predicting progress of disease in MDS. Clone size of trisomy 8 does not related to classification of MDS, but sole +8 is seems to see in MDS-RA frequently.
Chromosomes, Human, Pair 8
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genetics
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Humans
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In Situ Hybridization, Fluorescence
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Karyotyping
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Myelodysplastic Syndromes
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genetics
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pathology
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Reproducibility of Results
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Sensitivity and Specificity
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Trisomy
9.Slowing progression of chronic allograft nephropathy by conversion from cyclosporin A to tacrolimus.
Long-Kai PENG ; Xu-Biao XIE ; Feng-Hua PENG ; Yu WANG ; Yi JIANG ; Gong-Bin LAN ; Chun-Hua FANG ; Man-Hua NIE
Journal of Central South University(Medical Sciences) 2007;32(1):59-62
OBJECTIVE:
To investigate the feasibility and safety of substituting tacrolimus(FK506) for cyclosporin A(CsA) on delaying the pace of renal dysfunction in patients with biopsy-proven chronic allograft nephropathy(CAN).
METHODS:
Seventy-three renal transplantation patients with CAN proved by allograft biopsy were collected in this study. Patients were randomly divided into 2 groups. Patients were either converted to FK506(FK506 group, n=43) or remained on their initial CsA-based immunosuppression(CsA group, n=30). The clinical data at study entry and after 12 months including blood urea nitrogen(BUN), serum creatinine(SCr), glomerular filtration rate(GFR), 24-hour urine protein excretion, serum total cholesterol(TC), triglyceride(TG), low density lipoprotein(LDL) and the side effects of calcineurin inhibitors were monitored during a follow-up of over 12 months.
RESULTS:
Twelve months later, the level of SCr was statistically reduced and GFR levels were obviously elevated in the FK506 group as compared with CsA group [(194.8+/-42.5)micromol/L vs. (245.4+/-52.8)micromol/L and (50.14+/-3.92)mL/(min.1.73 m(2)) vs. (40.58+/-2.49)mL/(min.1.73 m2), P<0.01]. Quantity of 24-hour urine protein excretion in the FK506 group was (2.0+/-0.5)g which is significantly lower than (3.9+/-0.7)g in the CsA group(P<0.01). TC, TG, and LDL levels remained unchanged in the CsA group, while those were statistically reduced in the FK506 group respectively [(5.19+/-0.73)mmol/L vs. (6.94+/-1.37)mmol/L, (1.86+/-0.84)mmol/L vs. (3.14+/-1.38)mmol/L, (3.03+/-0.71)mmol/L vs. (3.82+/-0.89)mmol/L, P<0.01]. Tremor obviously increased (P<0.01) and hypertension obviously decreased (P<0.05) in the FK506 group compared with the CsA group.
CONCLUSION
FK506 treatment can greatly improve the proteinuria and hyperlipidemia. Conversion from CsA to FK506 is an effective and safe alternative therapy for delaying the progression of renal dysfunction induced by CAN.
Adult
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Aged
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Cholesterol
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blood
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Creatinine
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blood
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Cyclosporine
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therapeutic use
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Female
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Follow-Up Studies
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Glomerular Filtration Rate
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Graft Rejection
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complications
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Humans
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Immunosuppressive Agents
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therapeutic use
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Kidney Failure, Chronic
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blood
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drug therapy
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etiology
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Kidney Transplantation
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adverse effects
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Lipoproteins, LDL
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blood
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Male
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Middle Aged
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Tacrolimus
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therapeutic use
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Treatment Outcome
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Triglycerides
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blood
10.Postoperative changes in 25-hydroxy vitamin D and parathyroid hormone levels in obese patients
Xinyu CAO ; Zhe WANG ; Nianrong ZHANG ; Wen ZHANG ; Biao ZHOU ; Yuntao NIE ; Siqi WANG ; Hua MENG
Chinese Journal of Clinical Nutrition 2022;30(4):235-242
Objective:To investigate the postoperative changes in levels of 25-hydroxy vitamin D (25-[OH]D], parathyroid hormone (PTH) and other relevant biomarkers in obese patients receiving metabolic surgery and analyze the dynamic changes in relevant biomarkers in the short term (after 3-6 months) and the long term (after 12-24 months).Methods:A total of 96 obese patients who underwent metabolic surgery and received follow-up examinations from January 2018 to January 2020 were included. Baseline and postoperative data were collected, including anthropometric data (height, weight, neck circumference, waistline and hipline) and laboratory test results (fasting glucose, glycated hemoglobin, 25-[OH)D, PTH, serum calcium and serum phosphorus). Body mass index (BMI) and waist-hip ratio were calculated. The anthropometric data were analyzed by repeated measures analysis of variance and laboratory data were compared between groups using t test and Kruskal-Wallis test. Results:96 patients (33 males and 63 females) were included, of whom 49 were complicated with diabetes. Prior to surgery, 79 (79.17%) of the patients had 25-(OH)D deficiency (< 20 μg/L), 16 (16.67%) had 25-(OH)D insufficiency (≥ 20 μg/L and < 30 μg/L) and 23 (23.96%) had high PTH levels (> 70 ng/L). After the surgery, 25-(OH)D level was transiently increased in the short term ( P = 0.01) but declined thereafter in the long term ( P < 0.01) to levels lower than baseline ( P = 0.023). Long-term PTH level was higher than baseline ( P = 0.012), with 11 patients showing PTH levels higher than normal (> 70 ng/L). Serum phosphorus level was increased in both the short term and the long term ( P < 0.01). Conclusions:Obese patients have 25-(OH)D deficiency/insufficiency before metabolic surgery and experience further decrease in the long term after surgery, despite a transient increase. Secondary increase in PTH level occurs in some of the patients after surgery. Long-term nutritional supplements and comprehensive nutritional management play important roles in postoperative management of obese patient.