1.Effect of miRNA-5193 on the sensitivity of cervical cancer Caski cells to cisplatin
Guangxia WANG ; Shasha SHAO ; Lixin DONG
Cancer Research and Clinic 2021;33(3):161-167
Objective:To investigate the effect and mechanism of miRNA-5193 (miR-5193) on the sensitivity of cervical cancer Caski cells to cisplatin.Methods:The expression of miR-5193 in cervical cancer cell lines C33A, SiHa, Caski and normal cervical cell line Ect1/E6E7 were determined by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). Caski cells were divided into control group (no transfection, normally cultured), miR-5193-negative control (miR-NC) group (transfected with miR-NC mimic), miR-5193 group (transfected with miR-5193 mimic), miR-NC+cisplatin group (transfected with miR-NC mimic and treated with 10 μg/ml cisplatin), miR-5193+cisplatin group (transfected with miR-5193 mimic and treated with 10 μg/ml cisplatin), miR-5193+cisplatin+NC group (cotransfected with Foxp3-negative control vector and miR-5193 mimic, and treated with 10 μg/ml cisplatin), and miR-5193+cisplatin+Foxp3 group (cotransfected with Foxp3 overexpression vector and miR-5193 mimic, and treated with 10 μg/ml cisplatin). Proliferation was detected by methyl thiazolyl tetrazolium (MTT), cell cycle was detected by PI single staining method, cell apoptosis was detected by Annexin V-FITC/PI double staining method, and expressions of CDK2, p27 and C-caspase-3 proteins in cells were detected by Western blot. Bioinformatics software was used to predict miR-5193 target genes, and the luciferase reporting system was used to identify the targeting relationship.Results:The relative expression of miR-5193 in cervical cancer C33A, SiHa and Caski cells was lower than that in normal cervical Ect1/E6E7 cells (0.56±0.06, 0.41±0.03, 0.23±0.02 vs. 1.00±0.10, all P < 0.05). Compared with the control group and miR-NC group, the cell proliferation activity (absorbance value) in miR-5193, miR-NC+cisplatin and miR-5193+cisplatin groups decreased (0.58±0.06, 0.59±0.07 vs. 0.38±0.04, 0.40±0.05, 0.23±0.02, all P < 0.05), the cell apoptosis rate increased [(2.5±0.2)%, (2.7±0.3)% vs. (12.6±1.2)%, (11.9±1.5)% , (18.9±1.7)%, all P < 0.05], and the proportion of cells in G 0/G 1 phase increased [(50.4±4.2)%, (51.3±6.3)% vs. (62.3±3.2)%, (61.9±5.8)%, (71.4±5.4)%, all P < 0.05]. The expression levels of p27 and C-caspase-3 proteins increased, and the expression level of CDK2 protein decreased. The software predicted that the target gene of miR-5193 was Foxp3, which was confirmed by the luciferase reporting system. Compared with the miR-5193+cisplatin+NC group, the cell proliferation activity (absorbance value) in miR-5193+ cisplatin+Foxp3 group increased (0.24±0.03 vs. 0.65±0.05, t = 21.094, P < 0.01), the proportion of cells in G 0/G 1 phase decreased [(71.0±6.4)% vs. (60.3±4.1)%, t = 4.196, P < 0.01], the apoptosis rate of cells decreased [(19.6±1.6)% vs. (11.5±1.2)%, t = 11.880, P < 0.01], the expression levels of p27 and C-caspase-3 proteins in cells decreased, and the expression levels of CDK2 and Foxp3 proteins increased. Conclusion:The miR-5193 may increase the sensitivity of cervical cancer Caski cells to cisplatin in vitro by targeted inhibition of the Foxp3 gene.
2.Study on Optimum Conditions of Isolation and Purification of Specific Toxin Fractions Produced by Exserohilum turcicum
Shao-Xin WANG ; Jin-Gao DONG ;
Microbiology 1992;0(02):-
Five fractions,Ⅰ(R_f0.06)、Ⅱ(R_f0.21)、Ⅲ(R_f0.45)、Ⅳ(R_f0.60)、Ⅴ(R_f0.75)have been obtained after HT-toxin from race 1 of Exserohilum turcicum 99-2 isolated by TLC. In all of these fractions,only fractions Ⅱ had specific toxicity to the corn leaves with Ht1 gene. Then fractions Ⅱ-1、Ⅱ-2、Ⅱ-3 were isolated from fractions Ⅱ by HPLC purification,and the bioassay result showed only fraction Ⅱ-3 was toxigentic to corn leaves with Ht1 gene but non-toxigentic to corn leaves without Ht1 gene. Fractions Ⅱ-1、Ⅱ-2 and Ⅱ-3 were scanned by UV-Vis spectrophotometer. It was shown that the fractions Ⅱ-1 and Ⅱ-2 had analogous spectrum,and especially the fraction Ⅱ-3 had a special peak at 300nm.
3.Different scoring systems to evaluate the prognosis of Fournier's gangrene: A comparative study.
Xiao-dong ZHU ; Fei DING ; Guo-dong WANG ; Qiang SHAO
National Journal of Andrology 2015;21(8):720-723
OBJECTIVETo sum up the experience in diagnosis and treatment of Fournier's gangrene and find an optimal evaluation tool for its prognosis by comparing currently used prognostic scoring systems.
METHODSWe retrospectively analyzed 16 cases of Fournier's gangrene diagnosed and surgically treated in our hospital between 2004 and 2012. Using Fournier's Gangrene Severity Index (FGSI), Uludag Fournier's Gangrene Severity Index (UFGSI), Age-Adjusted Charlson Comorbidity Index (ACCI), and Surgical Apgar Score (sAPGAR) , we obtained the prognostic scores of the patients and made comparisons among different scoring systems.
RESULTSFGSI, UFGSI, ACCI, and sAPGAR were all clinically used scoring systems. Statistically significant differences were found in the scores of ACCI and UFGSI but not in those of FGSI and sAPGAR between the death and survival groups, with the maximum area under the ROC curve and minimum standard error for the ACCI score.
CONCLUSIONBoth ACCI and UFGSI are useful for evaluating the prognosis of Fournier's gangrene. However, ACCI is even better for its higher sensitivity and specificity and easier clinical collection.
Age Factors ; Aged ; Fournier Gangrene ; diagnosis ; mortality ; surgery ; Humans ; Prognosis ; Retrospective Studies ; Sensitivity and Specificity ; Severity of Illness Index
4.Effect of Carvedilol and Bisoprolol on the Changes of Heart Type Fatty Acid Binding Protein and Brain Natri-uretic Peptide in Patients with Congestive Heart Failure
Haitao ZHANG ; Hui SHAO ; Xinhong WANG ; Qiuli DONG ; Zhongming WANG
China Pharmacy 2015;(32):4532-4533,4534
OBJECTIVE:To observe the changes of heart type fatty binding protein (H-FABP) and brain natriuretic peptide (BNP)when carvedilol and bisoprolol in the treatment of elderly patients with chronic heart failure(CHF). METHODS:110 CHF patients were randomly assigned into carvedilol group and bisoprolol group (n=55). Carvedilol group was given carvedilol 3.125 mg,bid,doubling dose every 2 weeks to reach target dose 25mg or maximum tolerant dose;bisoprolol group was given bisoprolol 1.25 mg,qd,doubling dose every 2 weeks to reach target dose 10 mg or maximum tolerant dose. Treatment course lasted for 6 months. Clinical effective rate of 2 groups were observed after treatment. Echocardiography was used to measure the changes of LVEDD,LVEF,SBP and DBP before and after treatment. The plasma concentration of BNP and H-FABP were measured by radio-immunoassay. RESULTS:After treatment,total effective rate of carvedilol group(96.4%)was higher than that of bisoprolol group (89.2%),with statistical significance(P<0.05). Compared with before treatment,the levels of LVEDD,LVEF,BNP,H-FABP, SBP and DBP in 2 groups were all decreased significantly after treatment,with statistical significance (P<0.01 or P<0.05). CON-CLUSIONS:Both carvedilol and bisoprolol can significantly improve cardiac function,but carvedilol efficacy is more significant. At the same time,significant decrease of H-FABP before and after treatment indicate that H-FABP can be used as a reliable index of heart failure treatment effect.
5.Testing method of human body's current threshold for perception based on EEG analysis.
Xiaofei WANG ; Lijuan SHI ; Dong LI ; Xu ZHAO ; Haiming SHAO
Journal of Biomedical Engineering 2014;31(1):13-17
Electric and electronic products are required to pass through the certification on electrical safety performance before entering into the market in order to reduce electrical shock and electrical fire so as to protect the safety of people and property. The leakage current is the most important factor in testing the electrical safety performance and the test theory is based on the perception current effect and threshold. The traditional method testing the current threshold for perception only depends on the sensing of the human body and is affected by psychological factors. Some authors filter the effect of subjective sensation by using physiological and psychological statistical algorithm in recent years and the reliability and consistency of the experiment data are improved. We established an experiment system of testing the human hody's current threshold for perception based on EEG feature analysis, and obtained 967 groups of data. We used wavelet packet analysis to detect a wave from EEG, and used FFT to do spectral analysis on alpha wave before and after the current flew through the human body. The study has shown that about 97.72% alpha wave energy changes significantly when electrical stimulation occurs. It is well proved that when the EEG feature identification is applied to test the human body current threshold for perception, and meanwhile alpha wave energy change and human body sensing are used together to confirm if the current flowing through the human body reaches the perception threshold, the measurement of the human body current threshold for perception could be carried out objectively and accurately.
Algorithms
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Electric Stimulation
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Electroencephalography
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Humans
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Reproducibility of Results
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Sensory Thresholds
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physiology
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Wavelet Analysis
6.Effect of basic fibroblast growth factor on the proliferation and differentiation of non-adherent stromal precursor cells
Junfei SHAO ; Caiping MAO ; Mingchen WANG ; Ningzheng DONG
Chinese Pharmacological Bulletin 1986;0(05):-
AIM To investigate the actions of basic fibroblast growth factor (bFGF) on the proliferation and differentiation of the non-adherent stromal precursor (NASP) cells. METHODS The osteogenic potential of the bone marrow stromal cells (BMSC) isolated from Wistar rats were cultured in the absence or presence of bFGF. After ALP cytochemistry, the colonies were counted by image analysis. The cells cultured in petri dishes were stained by the avidin-biotin-peroxidase complex (ABC) immunoperoxidase technique for collagen type Ⅰ, proliferating cell nuclear antigen ( PCNA ) and double staining for calcium and osteocalcin. RESULTS Many stromal precursor cells are present in bone marrow in a non-adherent form. bFGF not only stimulated the proliferation of NASP cells, but also enhanced the differentiation of NASP cells into osteoblasts. CONCLUSIONS NASP cells are possibly the main targets of the anabolic action of bFGF which may play an important role in fracture healing and bone formation.
7.Traditional Chinese medicine Rhizoma curcumae activates PXR and affects cytochrome P450 3A in the rat liver
Jingwei SHAO ; Haiyan DONG ; Tao WANG ; Yanghao GUO
Chinese Pharmacological Bulletin 1986;0(04):-
Aim To investigate whether Rhizoma curcumaecould induce pregnane X receptor(PXR)-mediated transcriptional expression of CYP3A4 and study the modulatory effects on the enzyme activity and mRNA expression of CYP3A in the rat liver.Methods Transient cotransfection reporter gene assays were performed in HepG2 cells;the rat liver microsomal cytochrome P450 and CYP3A isoenzyme-erythromycin N-demethylase(ERD)activities were determined by UV chromatography;the mRNA expression level of CYP3A was detected by reverse transcriptase-polymerase chain reaction(RT-PCR).Results In vitroinvestigation showed Rhizoma curcumaeand its four selected constituents could induce the CYP3A4 transcriptional expression by activating PXR.In vivoinvestigation showed the CYP450 content of liver microsomes and enzyme activity of CYP3A were markedly increased and induced by Rhizoma curcumaeextract;at the mRNA level, the expression of CYP3A1 and CYP3A2 gene were markedly induced by Rhizoma curcumaeextract.Conclusions Rhizoma curcumaeand its four selected constituents could induce the expression of the CYP3A4 gene transcriptional expression through activating PXR;Rhizoma curcumaeextract could increase and induce the enzyme activity and mRNA expression of CYP3A.
8.MicroRNA differential expression in liver cirrhosis rats undergoing human umbilical cord mesenchymal stem cell transplantation
Xiangzhong LIU ; Zhiqiang ZOU ; Guiqiang WANG ; Dong LI ; Zhiying SHAO
Chinese Journal of Tissue Engineering Research 2015;(23):3674-3680
BACKGROUND:Human umbilical cord mesenchymal stem cels (hUC-MSCs) can obviously relieve liver cirrhosis, and thereby repair liver injury. However, the molecular mechanism of hUC-MSCs therapy for liver cirrhosis is limited at present, and especialy the non-coding RNA regulation of hepatic gene changes has not been detailed. OBJECTIVE:To investigate the changes of microRNA after hUC-MSCs therapy in rats with liver cirrhosis. METHODS:Liver cirrhosis models were established in rats using carbon tetrachloride subcutaneous injection plus oral administration of alcohol. At 8 weeks after modeling, hUC-MSCs were injectedvia the tail vein once a week for 4 consecutive weeks. At 1 week after the last injection, rat liver tissues were colected for paraffin embedding. Liver RNA was extracted for gene chip analysis. Blood samples were colected and analyzed using an automatic biochemical analyzer to detect the changes of liver function. RESULTS AND CONCLUSION:Alanine aminotransferase, aspartate aminotransferase and gamma-glutamyl transpeptidase were improved significantly after hUC-MSCs therapy. Fat lesions and necrosis of hepatocytes were significantly reduced. MicroRNA expression microarray hybridization analysis and PCR results showed that rno-miR-369-5p, rno-miR-3584-5p and rno-miR-153* were down-regulated during modeling and increased after hUC-MSCs therapy. And rno-miR-93, rno-miR-199a-3p, rno-miR-195, rno-let-7a and rno-miR-19a were firstly up-regulated in the process of modeling and then down-regulated obviously after hUC-MSCs therapy. These results suggest that hUC-MSCs may reverse liver cirrhosis and liver cel damage through up-regulation of rno-miR-369-5p, rno-miR-3584-5p and rno-miR-153*, and down-regulation of rno-miR-93, rno-miR-199a-3p, rno-miR-195, rno-let-7a and rno-miR-19a.
9.Analysis of the color ultrasonography of thyroid disease in adult people with neck discomfort
Hongyan WANG ; Xiaohui SHAO ; Xiaoqiu DONG ; Meina LIU
Chinese Journal of Endemiology 2014;33(3):342-345
Objective To investigate the current situation of thyroid disease by analyzing retrospectively the color poppler ultrasonography data of neck discomfort in Heilongjiang Province.Methods A total of 13 020 cases (18-93 years old) of thyroid ultrasound examination data collected from September 2009 to October 2012 at the Department of Ultrasound,Fourth Affiliated Hospital of Harbin Medical University,were retrospectively analyzed.According to the sonographic features of thyroid (thyroid morphology,size,echo characteristics,blood flow,nodular location,number,calcification,etc),combined with ultrasound diagnosis,the relationships between gender and disease,age and disease,ultrasound diagnosis and disease classification,gender,number of nodules and benign and malignant were analyzed.Results Among the 13 020 cases,524 cases was excluded.There were 2 291 male cases; abnormalities were detected in 1 679 cases,and the abnormal ratio was 73.29%; there were 10 205 female cases; 7 946 cases were abnormal,and the abnormal ratio was 77.86%.The proportion of abnormal thyroid cases of the total number of female was higher than that of male,and the ratio of male and female was 1.00 ∶ 4.73 (1 679∶7 946); gender differences were statistically significant(x2 =18.476,P < 0.01).There were nodular type 5 018 cases,diffuse type 2 012 cases and mixed type 2 603 cases in sonographic findings.The nodular type on ultrasound images showed mainly nodular goiter(79.21%,3 975/5 018).The diffuse type on ultrasonic images showed mainly Hashimoto Thyroiditis(59.24%,1 192/2 012).The mixed cases on ultrasound images showed mainly nodular goiter (89.83%,2 331/2 595).Ultrasound diagnostic results showed that nodular goiter accounted for 65.52% (6 306/9 625) and Hashimoto Thyroiditis accounted for 17.97% (1 730/9 625).Proportion of thyroid disease increased gradually with age,reached a peak [29.93%(2 557/8 544) and 28.84% (2 464/8 544)] between the ages of 41-50 and 51-60 years old.The proportion gradually decreased into[14.17%(1 211/8 544),7.26%(620/8 544)] between the age of 61-71 and > 71 years old.The incidence differences of malignant nodules between different age groups were statistically significant (x2 =407.796,P < 0.01).Among malignant nodules,solitary nodule accounted for 95.76% (113/118) ; multiple nodules accounted for 4.24% (5/118),and there were more solitary nodule than multiple nodules(x2 =15.286,P < 0.01).About malignant solitary nodules,women accounted for 87.61%(99/113); men accounted for 12.39% (14/113),and women's were significantly higher than man's(x2 =360.960,P < 0.01).Correlation analysis showed that the results of ultrasounddiagnosisofvarious thyroid diseases were highly correlated with the ultrasonic types of diseases(r =0.139 99,P < 0.01).Conclusions Nodular goiter and Hashimoto Thyroiditis are the most common adult neck discomfort diagnosed by ultrasound in Heilongjiang Province.The high-risk age of thyroid disease is between 41-60 years old.Thyroid malignant is more common in single nodular,and the number of female patients is significantly higher than that of men.
10.Clinical analysis of different hysterectomy approaches on 710 cases
Li MA ; Ailing SHAO ; Lixiao WANG ; Jianmin DONG
Chinese Journal of Primary Medicine and Pharmacy 2008;15(10):1606-1607
Objective To compare the clinical effects of total abdominal hysterectomy(TAH), laparoscopic vaginal hysterectomy(LAVH) and transvaginal hysterectomy(TVH). Methods A retrespective analysis was conduc-ted in 710 women underwent different hysterectomy approaches,includlng 397 patients underdoing TAH,176 underdo-ing LAVH and 137 treated with TVH. Results Uterine volume, operative pelvis, pelvic adhesion were main factors infulencing different operative styles for hysterectomy ( P<0.05 ). Compared with TAH, LAVH, TVH, the operation time,blood loss ,operation complicafion and recovery time of intestinal function were significantly difference(P<0.05). Conclusion Uterine volume, pelvic adhesion, operative pelvis were major factors influencing different operative styles for hysterectomy. The clinical efficacy of TVH is best,which is less time, minimal invasion, shorter recovery time and lower cost. The application of LAVH provides a promising microinvasive approach for vaginal hysterectomy which can make operation easy, visual field clear and is worthy to be advocated too.