1.Expression and Purification of Enterovirus Type 71 Polyprotein P1 using Pichia pastoris system
Xue HAN ; Xiaoling YING ; Hao HUANG ; Shili ZHOU ; Qi HUANG
Virologica Sinica 2012;27(4):254-258
Enterovirus type 71(EV71) causes severe hand-foot-and-mouth disease(HFMD) resulting in hundreds of deaths of children every year; However,currently,there is no effective treatment for EV71.In this study,the EV71 poly-protein(EV71-P1 protein) gene was processed and cloned into the eukaryotic expression vector pPIC9k and then expressed in Pichia pastoris strain GS115.The EV71 P1 pretein with a molecular weight of 100 kD was produced and secreted into the medium.The soluble EV71 P1 protein was purified by column chromatography with a recovery efficiency of 70%.The result of the immunological analysis showed that the EV71 P1 protein had excellent immunogenicity and could stimulate the production of EV71-VP1 IgG antibody in injected rabbits.We suggest that EV71-P1 protein is an ideal candidate for an EV71 vaccine to prevent EV71 infection.
3.THE POSSIBLE MUTAGENESIS OF SEVERAL MALE ANTIFERTILITY AGENTS IN SPERM HEAD ANOMALIES
Weisan YE ; Yanchun DENG ; Yuling HUANG ; Yuhua HAN ; Shepu XUE
Acta Anatomica Sinica 1955;0(03):-
Mutagenesis of several male contraceptives in sperm bead anomalies was investigated. Results show that glycosides of tripterygium wilfordii hook (GTW) and its monomer T13, microwave induce sperm head anomalies. However, gossypol and monomer T4 and GTW do not induce sperm head anomalies. Adult male mice and rats were given orally GTW, monomer T4, T13 and gossypol. These chemical agents were delivered in 1% methylcellulose. Result indicated that frequency of abnormal sperm heads in GTW, T13 groups were significantly increased, while frequency of abnormal sperm heads in T4 and gossypol-treated animals were similar to that of normal controls. When male mice were exposed to microwaves of 0.5 kW for 1-2 min, for five weeks abnormal shape of spermatozoa could be found.
4.Differentiation of small breast nodules using breast imaging reporting and data system-ultrasound
Shuai, ZHOU ; Min-xia, HU ; Qiang, ZHU ; Hui-lian, HUANG ; Xue-yu, RONG ; Han-xue, ZHAO ; Yu, CHEN
Chinese Journal of Medical Ultrasound (Electronic Edition) 2013;(8):662-668
Objective To study the interobserver variabilities and the differential diagnosis value of Breast Imaging Reporting and Data System-Ultrasound (BI-RADS-US) lexicon for small ( ≤ 2 cm) breast nodules. Methods Between January 2009 and December 2011, 289 patients with small (≤2 cm) breast nodules (n=317) were included. According to sizes, the lesions were divided into two groups, i.e., 0-1 cm (n=160) group and 1-2 cm (n=157)group. Each lesion was described independently by 3 radiologists using BI-RADS-US lexicon. Interobserver variabilities were assessed by Kappa test. Chi-square test was used to compare the frequency difference of the descriptors between malignant and benign lesions. Sensitivity, speciifcity, accuracy, positive predictive value and negtive predictive value were calculated. Results (1)Moderate agreements were obtained for lesion shape, orientation, margin, echo pattern, surrounding tissue and calciifcations (κ=0.44, 0.57, 0.48, 0.43, 0.51 and 0.57) in 0-1 cm group. Substantial agreements were obtained for lesion shape, orientation, margin and echo pattern (κ=0.65, 0.61, 0.64 and 0.63) in 1-2 cm group. (2)Irregular shape, non-parallel orientation, non-circumscribed margin, echogenic halo and microcalciifcations were more frequently found in malignant nodules than in benign nodules in 0-1 cm group [52.3% (34/65) vs 20.0% (19/95), 38.5%(25/65) vs 13.7%(13/95), 75.4%(49/65) vs 32.6%(31/95), 18.6%(12/65) vs 0 (0/95) and 10.8%(7/65) vs 2.1%(2/95);χ2=18.19, 13.08, 28.22, 16.39 and 3.95;P=0.000, 0.000, 0.000, 0.000 and 0.047]. Similarly, irregular shape, non-parallel orientation, non-circumscribed margin, echogenic halo, shadowing, changes of Cooper′s ligament and microcalciifcations were signiifcantly more frequent found in malignant nodules than in benign nodules in 1-2 cm group [74.2%(49/66) vs 12.1%(11/91), 36.3%(24/66) vs 5.5%(5/91), 93.9%(62/66) vs 22.0%(20/91), 37.9%(25/66) vs 3.3%(3/91), 30.3%(20/66) vs 7.7%(7/91), 15.2%(10/66) vs 0 (0/91) and 16.7%(11/66) vs 4.4%(4/91);χ2=62.59, 24.21, 79.40, 31.22, 13.73, 12.30 and 6.67;P=0.000, 0.000, 0.000, 0.000, 0.000, 0.000 and 0.010]. (3)In both groups, a good sensitivity was demonstrated (75.4%&93.9%) when using the non-circumscribed margin as a criterion for malignancy, and high speciifcity was achieved in two groups (80.0%-100%and 87.9%-100%) when other descriptors including irregular shape, non-parallel orientation, echogenic halo, shadowing, changes of Cooper′s ligament and microcalciifcations were used as differentiation criteria. Conclusions Good interobserver agreement can be achieved using the BI-RADS-US lexicon in the diagnosis of small breast nodules. Non-circumscribed margin are proved as the most valuable sign for screening malignant breast lesions ≤ 2 cm. High speciifcity was found for irregular shape, nonparallel orientation, echogenic halo, shadowing, Cooper′s ligament changes and microcalciifcations, which can help biopsy and preoperative diagnosis.
5.Myricetin attenuates renal fibrosis by activating Nrf2/HO-1 pathway to inhibit oxidative stress
Dong-xue LI ; Zhou HUANG ; Han-yu WANG ; Zhi-hao ZHANG ; Ning-hua TAN ; Xue-yang DENG
Acta Pharmaceutica Sinica 2024;59(2):359-367
This paper investigates the effect of myricetin (MYR) on renal fibrosis induced by unilateral ureteral obstruction (UUO) and common bile duct ligation (CBDL) in mice and its mechanism. The animal experiment has been approved by the Ethics Committee of China Pharmaceutical University (NO: 2022-10-020). Thirty-five ICR mice were divided into control, UUO, UUO+MYR, CBDL and CBDL+MYR groups. H&E and Masson staining were used to detect pathological changes in kidney tissues. Western blot (WB) was used to detect the expression of fibrosis-related proteins in renal tissue, and total superoxide dismutase (SOD) activity detection kit (WST-8) was used to detect the changes of total SOD in renal tissue of CBDL mice.
6.Lipopolysaccharide, TNFα, IL-6, dexamethasone, and insulin increase the expression of GPR54 in the MCF7 breasr cancer cell line.
Bing-kun HUANG ; Jiang-feng MAO ; Zhao SUN ; Qin HAN ; Min NIE ; Xue-yan WU
National Journal of Andrology 2015;21(7):587-592
OBJECTIVETo investigate the effects of different concentrations of lipopolysaccharide (LPS), tumor necrosis factor α (TNFα), interleukin-6 (IL-6), dexamethasone (Dex), and insulin on the mRNA and protein expressions of GPR54 in the MCF7 cell line in vitro.
METHODSMCF7 breasr cancer cells were cultured and treated with different concentrations of LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L). Those treated with culture fluid only served as controls. The mRNA and protein expressions of GPR54 were measured by real-time PCR and Western blot, respectively, after 6, 24, 48, and 72 hours of treatment.
RESULTSCompared with the blank con- trol, LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L) significantly increased the expressions of GPR54 mRNA (P < 0.05) and protein (P < 0.05).
CONCLUSIONLPS, TNFα, IL-6, Dex, and insulin evidently increase the expression of GPR54 in the MCF7 cell line, indicating their influence on the function of gonads by regulating the GPR54 level.
Blotting, Western ; Dexamethasone ; administration & dosage ; pharmacology ; Glucocorticoids ; administration & dosage ; pharmacology ; Gonads ; drug effects ; metabolism ; Humans ; Hypoglycemic Agents ; administration & dosage ; pharmacology ; Insulin ; administration & dosage ; pharmacology ; Interleukin-6 ; administration & dosage ; pharmacology ; Lipopolysaccharides ; administration & dosage ; pharmacology ; MCF-7 Cells ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, G-Protein-Coupled ; drug effects ; genetics ; metabolism ; Receptors, Kisspeptin-1 ; Time Factors ; Tumor Necrosis Factor-alpha ; administration & dosage ; pharmacology
7.Optimization of a Liquid Chip System for the Detection of Serum Biomarkers of Colorectal Cancer and Its Application
Hua-Ning ZHANG ; Xue-Qin GAO ; Jin-Xiang HAN ; Hai-Nan HUANG ;
China Biotechnology 2006;0(12):-
Liquid chip technology have been licensed to be used in clinic because of its advantage of high-throughput, high-sensitivity, good signal to noise ratio, reaction in liquid phase, convenient operation and short time consuming, etc. The optimization of a liquid chip system for the detection of serum biomarkers of colorectal tumour and initial application in the detection of CEA were studied. The optimized reaction conditions of liquid chip were determined through orthogonal design after it was prepared. The results showed that the consuming reaction time of the coated antibody and the antigen was 1hour. The microspheres, biotinylated detecion antibody and the consuming complexes and avidin-PE time of the microspheres and the biotinylated tested antibody was 1hour, 1hour and 15minutes respectively.the consuming time of the complexes and avidin-PE was fifteen minutes, The optimized dilution of the biotinylated tested detection antibody was 1∶300 and the optimized concentration of avidin-PE was 12?g/ml. Totally 55 clinical samples were detected by the liquid chip and by Enzyme-Linked Immunosorbent Assay (ELISA) simultaneously and the results of the two methods were compared. The results of the two methods showed good correlation between positive and negative samples but the detection limits and the dynamic ranges of the liquid chip method were more sensitive and wider than those of the ELISA. The multiple tumour biomarkers may be detected simultaneously and the time of clinical test and manpower requirements were reduced by the liquid chip method.
8.Analyses of four?-adrenergic receptor antagonists in the treatment of chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS)
Hong-Jun LI ; Han-Zhong LI ; Xue-Jun SHANG ; Yu-Feng HUANG ;
Chinese Journal of Urology 2001;0(06):-
Objective To investigate the efficacy and side effects of 4?-adrenergic receptor(?- AR)antagonists in the treatment of chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS).Meth- ods Totally,325 patients(mean age,33.2 year;disease history,2.4 years)with CP/CPPS were randomly divided into Prazosin(n=95),Terazosin(n=78),Phenoxybenzamine(n=27),and Doxazosin mesylate controlled release tablets(n=72)groups.Some antibiotics and medications were used for allopathy.In addi- tion,53 cases with no?-AR antagonist treatment served as control group.The efficacy and side effects in all the patients were observed and recorded.The chronic prostatitis symptom index(CPSI)was used to evaluate the efficacy.Results In control group,22(41.5%)patients responded well to the treatment( CPSI mark drop≥5)and 31(58.5%)failed to respond to the treatment(CPSI mark drop<5).In study group,199 (73.2%)patients responded well to the treatment,and 73(26.8%)failed.The difference in efficacy be- tween?-AR antagonists and placebo treatment was significant(P<0.01).In study group,specifically,the effective rate was 55.6% in Phenoxybenzamine,78.2% in Terazosin,76.4% in Doxazosin mesylate con- trolled release tablets,and 71.6% in Prazosin groups.The main side effects of?-AR antagonists were postur- al hypotension(a rate of 23.2% for Prazosin,17.9% for Terazosin,22.2% for Phenoxybenzamine,and 8.3% for Doxazosin mesylate controlled release tablets)and dysfunction of ejaculation(only in Phenoxy- benzamine group with a rate of 51.9%).The rates of withdrawing treatment due to side effects were in turn 18.5% of Phenoxybenzamine,7.4% of Prazosin,5.1% of Terazosin,and 0% of Doxazosin mesylate con- trolled release tablets.Conclusions As essential medications for the treatment of CP/CPPS,?-AR antag- onists can relieve the clinical symptoms(dropping NIH-CPSI mark significantly),but some side effects should be considered when some medications are selected.
9.Clinical significance of miR-143 and miR-145 expression in gastric gastrointestinal stromal tumor
Zhi LIU ; Shaoliang HAN ; Yingpeng HUANG ; Wenyi WU ; Junlin LI ; Xiangyang XUE ; Xian SHEN
Chinese Journal of General Surgery 2010;25(8):678-680
Objective To evaluate the role of miR-143, miR-145 in the development of gastric gastrointestinal stromal tumor. Methods The expression levels of miR-143 and miR-145 in 21 cases of gastric gastrointestinal stromal tumor and the matched non-tumor adjacent tissue specimens were examined by stem-loop real-time RT-PCR, and its correlation with clinicopathologic features of gastric gastrointestinal stromal tumor were analyzed. Results Expression level of miR-145 were significantly higher in tumor than adjacent normal tissues (P<0.01 ) and that with mitotic count ≥ 5/50HPF cases was significantly lower than that with mitotic count <5/50HPF cases (P=0.02). miR-145 expression in huge tumor (>10 cm)was significantly lower than that in the large tumor (5~10 cm) and small tumor (2~5 cm) (P=0.048).By Fletcher risk stratification system, miR-145 expression in high-risk cases was significantly lower than that in the intermediate-risk and low-risk cases (P=0.048). While the expression of miR-145 in low-risk group was significantly different compared to that in intermediate-risk group and high-risk group (P=0.01).There was no difference between the expressions of miR-143 in tumor and that in normal tissue(P=0.06).Conclusion In gastric gastrointestinal stromal tumor, MiR-145 expression is significantly higher in tumor than adjacent normal tissues. miR-145 is closely associated with tumor size. mitotic counts and Fletcher risk stratification system.
10.Relationship between genetic polymorphisms in nucleotide excision repair gene excision repair cross complement group 6, xeroderma pigmentosum group A and coal-burning-borne-arsenism
Xiaojing XU ; Aihua ZHANG ; Bing LIANG ; Xue HAN ; Xuexin DONG ; Xiaoxin HUANG
Chinese Journal of Endemiology 2014;33(1):15-20
Objective To investigate the relationship between genetic polymorphisms in nucleotide excision repair gene excision repair cross complementing group 6(ERCC6),xeroderma pigmentosum group A(XPA) and coal-burning-borne-arsenism in Guizhou Province.Method ERCC6 A3368G,ERCC6 C-6530G and XPA A23G gene polymorphisms were analyzed by polymerase chain reaction restriction fragment length polymorphism technique(PCR-RFLP) of 205 cases which were chosen as patients with arsenism and 187 residents as control group.Results The distributions of ERCC6 A3368G,ERCC6 C-6530G and XPA A23G in the case group were not statistically significant compared with those of the control group(x2 =3.209,2.963,3.335,all P > 0.05); individuals carrying G allelomorphic gene(AG + GG) had a lower risk than individuals carring A allelomorphic gene(ORadj =0.282,95%CI:0.126-0.628,P =0.002); relationship was not found between single genetic polymorphisms of ERCC6 C-6530G,XPA A23G and coal-burning-borne-arsenism; the risk of arsenism was decreased for individuals carrying the following five genotypes combination:ERCC6 A3368G(AG + GG) genotype and ERCC6 C-6530G CC genotype(ORadj =0.287,95%CI:0.087-0.946,P=0.040); ERCC6 A3368G(AG + GG) genotype and ERCC6 C-6530G(CG + GG) genotype (ORadj =0.226,95%CI:0.077-0.661,P =0.007); ERCC6 A3368G(AG + GG) genotype and XPA A23G AA genotype (ORadj =0.150,95%CI:0.038-0.596,P =0.007); ERCC6 A3368G (AG + GG) genotype and XPA A23G(AG + GG) genotype(ORadj =0.325,95%CI:0.118-0.897,P =0.030) ; ERCC6 C6530G (CG + GG) genotype and XPA A23G AA genotype (ORadj =0.397,95%CI:0.162-0.975,P=0.036).Conclusions Individuals carring ERCC6 A3368G (AG + GG) genotype have a low risk of arsenism.There are five genotypes combination of three gene polymorphisms in two genes,ERCC6 and XPA,which may reduce the risk of coal-burning-borne-arsenism.