2.The research on the expression and regulation of kallikrein 6 gene in human ovarian cancer cell
Fang ZHANG ; Ying-Jian CHEN ; Cheng-Jin HU ;
Chinese Journal of Laboratory Medicine 2001;0(04):-
Objective To investigate the effects of estrogen on the expression of KLK6 mRNA and protein in human ovarian cancer cell line HO8910.Methods HO8910 cells were incubated for 72 hours in two groups:the test group with 17-?E_2 at different concentration(10~(-10)、10~(-9)、10~(-8)、10~(-7)mol/L)and the control group with Ethanol.Real-time fluorensence quantitive RT-PCR and flow cytometry were used to measured the expression of KLK6 mRNA and protein in the two groups.The cell proliferation was measured by 3-(4,5-dimethylthiazol-z-yl)-2,5-dipheny tetrazolium blue(MTT)calorimetric assay,while the cell cycle was determined by flow cytometry.Results The expression ofKLK6 mRNA(3.83?0.41、4.14? 0.49、6.26?0.38、7.28?1.82)and protein(10.62?0.35,10.89?0.12、11.88?0.28、12.07?0.15) in the test group of H08910 cells(10~(-10)10~(-9)10~(-8)10~(-7)mol/L)was higher than that in the ethanol control group(P
3.Protein Kinase C Regulating Activation on Nuclear Factor-?B in T Lymphocyte in Children with Acute Idiopathic Thrombocytopenic Purpura
jin-cheng, XU ; hong, XIAO ; chang-lin, WU ; fang, LIU
Journal of Applied Clinical Pediatrics 2006;0(23):-
Objective To explore the role of protein kinase C(PKC) to regulate the activation of nuclear factor-?B(NF-?B)in T lymphocyte in children with acute idiopathic thrombocytopenic purpura(ITP).Methods Sterility peripheral blood was collected from acute ITP children(n=30)and healthy children(n=30).T lymphocytes were isolated and purified,and divided into 3 groups:control group,PMA group stimulated with PMA,PMA plus H-7 group stimulated with PMA and H-7.The expression of NF-?B and inhibitor protein-?B(I-?B)was detected by immunohistochemical staining and Western blot,respectively.Results The percentage of cells with active NF-?B was significantly higher and the expression level of I-?B was significantly lower in acute ITP PMA group than that in acute ITP control group and normal PMA group,respectively(all P
4.Tissue distribution of free anthraquinones in SD rats after orally administered extracts from raw and prepared rhubarbs.
Fang FANG ; Jiabo WANG ; Yanling ZHAO ; Cheng JIN ; Weijun KONG ; Haiping ZHAO ; Hongjuan WANG ; Xiaohe XIAO
Acta Pharmaceutica Sinica 2011;46(3):350-4
Rhubarb anthraquinone derivatives (AQs) have been documented to have both therapeutic and toxic effect on liver and kidney, leading to a complex puzzle to assess their benefits and risks. In this study, the tissue distributions of AQs in SD rats after orally administrated extracts of raw and prepared rhubarb were examined whether they undergo different uptake. The total rhubarb extract (14.49 g x kg(-1) of body weight per day od, counted on the quantity of crude material) was administrated orally for 12 weeks. The concentrations of the AQs in different tissues were quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The five major AQs, aloe-emodin, rhein, emodin, chrysophynol and physcion, could all be detected in the liver, kidney and spleen, while only rhein, aloe-emodin and emodin reached the quantitative limit. The tissue concentrations of AQs in raw rhubarb group were higher than that in steamed rhubarb group with rhein > emodin > aloe-emodin in the same tissue. On the whole, the tissue distribution of rhein was higher than that of emodin and aloe-emodin in liver, spleen and kidney. AQs could not be detected in those tissues after drug withdraw for 4 weeks, which suggested scarcely any accumulative toxicity of rhubarb. The result indicated that raw rhubarb had more tissue toxicity than steamed rhubarb and rhein may be one of the major poisonous ingredients. The results were concordant with the traditional Chinese medicine theory of toxicity-attenuating effect of processing.
5.The value of CT ribs unfolded algorithm in the diagnosis of rib fracture
Fang LU ; Xiaojun GE ; Cheng LI ; Liang JIN ; Xiu JIN ; Ming LI
Chinese Journal of Radiology 2017;51(9):695-698
Objective To evaluate the value of a CT ribs unfolding algorithm in the diagnosis of rib fracture. Methods Retrospective analysis of 180 patients who suffered chest trauma to do the chest or/and abdominal CT examination, and they also had the surgical or CT referral information. The images of these patients were postprocessed by software(Bone Reading software)and hand-drawn method(multi-point hand-painted CPR method). The rib fracture was observed and the time of reading was record. The diagnosis of fractures was confirmed by follow-up review or surgery. The fractures diagnosis sensitivity of the two post-treatment methods were measured, and the McNemar test was used to compare the difference between the software method and the hand-drawn method. Results Eight patients were excluded due to program failure, 172 cases were included in the study. Of the 172 patients, 63 patients suffered 259 fractures(178 ribs). The sensitivity of the software group was 91.7%(475/518), which was higher than that of the hand-painted group(86.3%, 447/518), the difference was statistically significant(P=0.005). The time of reading were (30.3 ± 3.3)and(173.2 ± 4.5)s, respectively, and the difference had statistically significant(P=0.001). Conclusion Compared to the traditional CPR method, the bone reading technique was used in patients with rib fractures during thoracic CT postprocessing can shorten the reading time and increase the sensitivity of the diagnosis.
6.The difference of T lymphocyte subsets between mild type and severe type in patients infected with the 2009 pandemic influenza A (H1N1) virus and its clinical significance
Jun WANG ; Jun JIN ; Fang HUANG ; Jianan HUANG ; Cheng JI ; Yueping SHEN ; Hua XU
Chinese Journal of Emergency Medicine 2011;20(2):193-197
Objective To study the changes of subgroups of peripheral blood T lymphocytes in the patients infected with the 2009 pandemic influenza A ( H1N1 ) virus of different severity type. Method A total of 66 patients infected by H1N1 evidenced by RT-PCR admitted from September 2009 to January 2010 were divided into three groups: mild type ( B group, n = 47 ), cured patients of severe and critical severe type ( C group, n = 14) and died patients ( D group, n =5), according to the severity and prognosis. A total of 20 healthy volunteers served as control group( A group). Peripheral blood lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count were detected by flow cytometry at the different time points. Fever duration and H1N1 virus negative time were compared. Statistical analysis were performed by using SAS version 9.13 software and the data were processed with ANOVA and SNK test. Results Lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count declined in the early period in all the groups, and there were significant differences compared with A group (P<0. 05), while rised with the clinical progression in group B and C,and those of C group were lower than B group ( P < 0.05 ), but those of D group were always low. Fever duration and H1N1 virus negative time were (4.4 ± 1.6) days vs. (4.4 ± 1. 4) days, ( 12.9 ± 3. 1 ) days vs.( 10.2 ± 2.6) days and ( 15.2 ± 7.3 ) days vs. ( 13.3 ± 2.9 ) days respectively, and there were significant differences among the three groups ( P < 0.05 ). Conclusions The cellular immune function was seriously damaged when patients were infected with H1N1. Further more, the changes of lymphocyte count, CD3+ , CD4+and CD8+ T lymphocyte count were tightly related with the degree of severity and prognosis. These findings can be used for clinical diagnosis and treatment.
7.Multispacer typing (MST) of spotted fever group rickettsiae isolated from humans and rats in Chengmai County, Hainan Province, China
Xueqin Cheng ; Yuming Jin ; Shijun Lao ; Changhe Huang ; Fang Huang ; Pengben Jia ; Lijuan Zhang
Tropical Medicine and Health 2014;():-
Spotted fever caused by spotted fever group rickettsiae (SFGR) is found throughout China. During 2007—2008, 28 human SFGR isolates and 34 rat SFGR isolates including 15 isolates from Rattus fulvescens, 5 isolates from R.edwardsi, 7 isolates from Callosciurus erythraeus roberti and 7 isolates from Dremomys rufigenis) were obtained from L929 cell culture. Previous research indicated that the 62 strains of SFGR mentioned above shared not only the same serophenotype but also 100% of identity sequences of 16S rRNA, gltA, ompA, groEL and 17KD, which enabled us to apply multispacer typing (MST) to the 62 SFGR isolates in the study. Six primer pairs, which were used for typing of Rickettsia rickettsii and Rickettsia conorii, were chosen, and the results exhibited greater nucleotide polymorphisms among the 62 isolates tested. A total of 48 distinct genotypes were identified. The dominant genotype, represented by h3 isolates, accounted for 21.7% (13/60) of the isolates tested, and the remaining 47 genotypes were all unique. Phylogenetic analysis showed that all the 48 genotypes could be classified in the same clade, while the genetically related strain, R.heilongjiangensis, was close but not the same as the cluster. We concluded that the genetically diverse of spotted fever group rickettsiae strains are endemic in Chengmai County, Hainan Province, China.
8.Multispacer Typing (MST) of Spotted Fever Group Rickettsiae Isolated from Humans and Rats in Chengmai County, Hainan Province, China
Xueqin Cheng ; Yuming Jin ; Shijun Lao ; Changhe Huang ; Fang Huang ; Pengben Jia ; Lijuan Zhang
Tropical Medicine and Health 2014;42(3):107-114
Spotted fever caused by spotted fever group rickettsiae (SFGR) is found throughout China. During 2007–2008, 28 human SFGR isolates and 34 rat SFGR isolates including 15 isolates from Rattus fulvescens, 5 isolates from R. edwardsi, 7 isolates from Callosciurus erythraeus roberti and 7 isolates from Dremomys rufigenis) were obtained from L929 cell culture. Previous research indicated that the 62 strains of SFGR mentioned above shared not only the same serophenotype but also 100% of identity sequences of 16S rRNA, gltA, ompA, groEL and 17KD, which enabled us to apply multispacer typing (MST) to the 62 SFGR isolates in the study. Six primer pairs, which were used for typing of Rickettsia rickettsii and Rickettsia conorii, were chosen, and the results exhibited greater nucleotide polymorphisms among the 62 isolates tested. A total of 48 distinct genotypes were identified. The dominant genotype, represented by h3 isolates, accounted for 21.7% (13/60) of the isolates tested, and the remaining 47 genotypes were all unique. Phylogenetic analysis showed that all the 48 genotypes could be classified in the same clade, while the genetically related strain, R. heilongjiangensis, was close but not the same as the cluster. We concluded that the genetically diverse of spotted fever group rickettsiae strains are endemic in Chengmai County, Hainan Province, China.
9.TRIB3 promotes lung cancer cell survival and inhibits apoptosis through NRF2 activation
Jiao-jiao YU ; Cheng ZHANG ; Yu-jin XIANG ; Zhuo-wei HU ; Bing CUI ; Fang HUA
Acta Pharmaceutica Sinica 2021;56(5):1352-1359
The nuclear transcription factor nuclear factor erythroid 2-related factor 2 (NRF2) plays a crucial role in maintaining cellular redox homeostasis. The aberrant NRF2 signaling confers enhanced antioxidant capacity, which is linked to tumor progression and therapeutic resistance. The current study investigates the biological effects and molecular mechanism of tribbles homolog 3 (TRIB3), a stress-induced protein, in regulating cell survival and apoptosis in lung cancer. This study first performed the RNA sequencing data analysis with 576 lung adenocarcinoma patients from the cancer genome atlas (TCGA) database. The NRF2- antioxidant response element (ARE) signature was enriched in patients with high TRIB3 expression. Dual-luciferase reporter assay and real-time quantitative polymerase chain reaction (PCR) were used to confirm the effect of TRIB3 on the kelch-like ECH-associated protein-1 (KEAP1)-NRF2 pathway. Abrogation of
10.Immunological effects of different adjuvants on HCV-DNA vaccine
Bo JIN ; Richard Yan-Hui Wang ; Liu-Fang CHENG ; Qi QIU ; Weiguo SHI ;
Chinese Journal of Immunology 1985;0(03):-
Objective:The immunological effects of HCV-DNA vaccine with different adjuvants were detected by ELISPOT in mice.Methods:Female BALB/c mice were primed with naked HCV-DNA, HCV-DNA encapsulated by liposome DDAB/EPC or DC-Chol/DOPE, HCV-DNA mixed with Montanide ISA 720 or aluminum hydroxide, respectively, and boosted twice accordingly in a four-week interval. Cytokine production by splenocytes was assessed by ELISPOT.Results:In most cases, splenocytes from mice vaccinated with DDAB/EPC liposome produced more IFN-?. These splenocytes also have significant higher IL-2 production compared with the other groups. In expansion with NS5b, splenocytes from alum group have significance in IL-4 production compared with other groups. The profile of cytokine production revealed that the INF-? overwhelmed IL-4 in naked DNA, DDAB/EPC, and DC-Chol/DOPE groups while IL-4 surmounted IFN-? in alum and Montanide groups.Conclusion:Encapsulation with liposome DDAB/EPC has the strongest adjuvant effect in inducing Th1 dominated immunity. Alum and Montanide can convert the Th1 nature of DNA vaccine to Th2-biased immunity.