1.Effect of tetrandine on gene expression of collagen type I, collagen type III and TGF-beta1 in scar tissue's of rabbits ear.
Xiao-Liang ZHOU ; De-Wu LIU ; Yuan-Gui MAO ; Jing LÜ
Chinese Journal of Plastic Surgery 2013;29(6):406-412
OBJECTIVETo observe the effect of tetrandine on gene expression of collagen type I, collagen type III, transformation growth factor-beta1 and to investigate the inhibitory effect of tetrandine on the scar tissue hyperplasia in rabbits' ears.
METHODSAfter the scar model was formed on the rabbits' ears, the rabbits were divided into 4 groups to receive intro-lesion injection with saline, or prednisolone (Pre) or tetrandrine in low concentration (L-Tet, 1.0 mg/ml) or tetrandrine in high concentration (H-Tet, 7.5 mg/ml). The morphological changes of scar tissue were observed. The changes of fibroblasts quantity and collagen expression were observed with HE and Masson staining. Immunohistochemical study was used to observe the expression level of collagen type I and collagen type III and TGF-beta1. Collagen type I and collagen type III and TGF-beta1, and signal factor Smad 3 mRNA were detected with RT-PCR.
RESULTS(1) 24 days after injury, all the wounds healed completely with formation of red, tough and hypertrophic scar. HE and Masson staining showed significant increase of fibroblasts and collagen density with irregularly arrangement. (2) Compared with that in saline group, the scar in other groups became softer, lighter and thinner, especially in H-Tet group. (3) HE and Masson staining shows the scar in Tet and Pre groups contained less fibroblasts and lower collagen dentsity with comparatively regular arrangement than that in saline group (P < 0.01), especially in H-Tet group. (4) According to the immunohistochemical study, the expression of collage type I and III and TGF-beta was positive in all the groups, but the positive rate and the ratio of collagen density I to III decreased in the order of saline, L-Tet, H-Tet and Pre groups (P < 0.01). (5) PT-PCR detection results showed that the amplification bands brightness of collagen type I and III and TGF-beta1 and signal molecular Smad 3 mRNA in scar tissue were obviously different. Compared with that in saline group, the expression of collagen type I and III and TGF-beta1 and Smad 3 mRNA decreased in Tet and Pre groups (P < 0.01). H-Tet group showed the most obvious reduce in the expression of type I collagen and TGF-beta1 and Smad 3 mRNA. Conclusions Tetrandine can significantly suppress the expression of collagen type I and collagen type III and TGF-beta1 on hypertrophic scar of rabbit ears, and reduce signal factor Smad 3 mRNA' s expression. It may be one of the important mechanism for its inhibitory effect on scar hyperplasia.
Animals ; Benzylisoquinolines ; pharmacology ; Cicatrix, Hypertrophic ; drug therapy ; genetics ; pathology ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Ear ; Fibroblasts ; Gene Expression ; Male ; RNA, Messenger ; metabolism ; Rabbits ; Smad3 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism
2.FGF-21 protects H9c2 cardiomyoblasts against hydrogen peroxide-induced oxidative stress injury.
Miao-Miao HAN ; Wen-Fei WANG ; Ming-Yao LIU ; De-Shan LI ; Bing ZHOU ; Yin-Hang YU ; Gui-Ping REN
Acta Pharmaceutica Sinica 2014;49(4):470-475
Fibroblast growth factor-21 (FGF-21) is an important metabolism regulator, however, whether FGF-21 has effects on cardiovascular remains unclear. In this study, H2O2-induced injury in H9c2 cells was used as a cell model, the anti-apoptosis potential and mechanism of FGF-21 against oxidative injury were evaluated by MTT assay, flow cytometry assay and real-time PCR. The results showed that FGF-21 could increase the cell survival of H2O2-induced injury in H9c2 cells and prevent H9c2 cells from oxidative stress-induced apoptosis. Furthermore, FGF-21 can elevate SOD activity and regulate Bcl-2/Bax expression in H9c2 cells. The results suggest that FGF-21 have protective effect against the H2O2-induced apoptosis in H9c2 cells.
Animals
;
Apoptosis
;
drug effects
;
Cell Proliferation
;
drug effects
;
Cells, Cultured
;
Fibroblast Growth Factors
;
pharmacology
;
Hydrogen Peroxide
;
toxicity
;
Malondialdehyde
;
metabolism
;
Myocytes, Cardiac
;
cytology
;
drug effects
;
metabolism
;
Oxidative Stress
;
drug effects
;
Protective Agents
;
pharmacology
;
Proto-Oncogene Proteins c-bcl-2
;
genetics
;
metabolism
;
RNA, Messenger
;
metabolism
;
Rats
;
Reactive Oxygen Species
;
metabolism
;
Superoxide Dismutase
;
metabolism
;
bcl-2-Associated X Protein
;
genetics
;
metabolism
3.Analysis of human brucellosis surveillance in Weinan city of Shaanxi province in 2009
Guang-de, LI ; Xing-you, HOU ; Hui-lan, QIAN ; Xin-gui, ZHANG ; Min, ZHOU ; Shi-hong, GUO
Chinese Journal of Endemiology 2011;30(2):205-207
Objective To survey and analyze characteristics of brucellosis epidemic in Weinan city of Shaanxi province for the purpose of setting up prevention and control measures for the disease. Methods According to "The Executing Plan for the Work of Surveying Brucellosis Disease in Shaanxi Province", 35 villages(towns) of designated monitoring locations and 24 villages (towns) of randomized monitoring locations in five countries of Weinan were chosen to survey brucellosis disease. The five countries were Chengcheng, Dali, Heyang, Tongguan and Hancheng. High risk populations with a history of contacting livestock and livestock products aged between 7 and 60 underwent clinical and serology examination[rose bengal plate agglutination test(RBPT) and standard tube agglutination test(SAT)]. All manipulation methods and judging standards were in accord with the "Diagnostic Standard for Brucellosis" (WS 269-2007). Results In the designated monitoring location, a total of 8664 people at high risk were investigated, among whom 1407 people were tested by RBPT test and 27 people were positive,the positive rate was 1.92%(27/1407); 27 people were tested by SAT test and 27 people were positive, the positive rate was 100% (27/27); 25 people were diagnosed and the diagnosis rate was 92.59%(25/27). In the randomized monitoring location, a total of 3464 people at high risk were investigated, among whom 411 people were tested by RBPT test and 3 people were positive, the positive rate was 0.73%(3/411 ), 3 people were tested by SAT test which were all positive and made a definite diagnosis. Twenty-eight new cases were made a definite diagnosis and its incidence was 2.06 in a hundred thousand(28/1 361 618). Conclusions The infection of human brucellosis in Weinan city stays at higher level. The governments should increase input for the monitoring,investigating and disinfecting to prevent the disease from increasing and outspreading.
4.Study on the mutations of ATP2C1 gene in patients with Harley-Harley disease
Xiao-Xiao YAN ; De-Ke JIANG ; Fu-Ren ZHANG ; Hong-Qing TIAN ; Gui-Zhi ZHOU ; Xian-Mei LU ; Long YU
Chinese Journal of Dermatology 1994;0(06):-
Objective To detect the mutations of ATP2C1 gene in patients with Hailey-Hailey dis- ease (HHD).Methods PCR and direct sequencing were performed in 17 patients and 120 healthy controls to screen the mutations in the exons of ATP2C1 gene.Results Eight mutations were identified in nine probands, including three deletion mutations (nt1464-1487 del/nt1462-1485del,1523delAT,2375delTTGT),three splice site mutations (360—2A→G,1415—2A→T,2243+2T→C) and two missence mutations (C920T and G1942T).None of the above mutations was found in the controls.Conclusion Eight specific novel mutations were identified in nine probands of HHD,which could be causative factors of the disease.
5.Phenylpropanoids and phenylethanol from flowers of Rosa rugosa.
Yue-de WANG ; Kun ZHOU ; Wei DONG ; Gui-you LIU ; Li-mei LI ; Jie LOU ; Qiu-fen HU ; Yan-qing YE ; Min ZHOU
China Journal of Chinese Materia Medica 2015;40(20):3998-4001
A new phenylpropanoid (1), together with seven known ones (2-8), has been isolated from the flowers of Rosa rugosa collected from Shanxi province by using various chromatographic techniques. Compound 1 is a new compound, and it displayed cytotoxicity against NB4, SH-SY5Y, PC3, A549 and MCF7 cell lines with IC₅₀ values of 8.2, 6.2, 4.3, 2.8, and 9.6 µmol · L⁻¹ respectively.
Cell Line
;
Cell Survival
;
drug effects
;
Drugs, Chinese Herbal
;
chemistry
;
isolation & purification
;
pharmacology
;
Flowers
;
chemistry
;
Humans
;
Molecular Structure
;
Phenylethyl Alcohol
;
chemistry
;
isolation & purification
;
pharmacology
;
Rosa
;
chemistry
;
Spectrometry, Mass, Electrospray Ionization
6.Study on age and education level and their relationship with fall-related injuries in Shanghai, China.
Yan Hong LI ; Gui Xiang SONG ; Yan YU ; De Ding ZHOU ; Hong Wei ZHANG
Biomedical and Environmental Sciences 2013;26(2):79-86
OBJECTIVETo study age and educational level and their relationship with fall-related injuries in Shanghai and to analyze the relevant costs.
METHODSMultistage cluster sampling was used for the selection of participants and standardized questionnaires were used for the information collection in 2006. Information on cases and deaths caused by fall-related injuries were obtained from 494 hospitals as well as from the mortality registry systems from 2001 till 2010.
RESULTSOf 45 857 participates, 674 suffered from fall-related injuries with the largest proportion among all injuries. The fall-related mortality increased from 10.63 per 100 000 in 2001 to 14.11 per 100 000 in 2010. The under-five mortality rate was the highest among children aged 0-14 years. Mortality increased dramatically among those aged 55 or above for the female and aged 60 or older for the male. Individuals with an educational level under the primary school were more likely to suffer fall-related injuries, accounting for 72.66% of all deaths and 49.24% of nonfatal cases respectively. The annual burden of fall-related injuries equated to 25.90% of the share of GDP for the healthcare, social security and welfare industries in 2006.
CONCLUSIONFall-related injuries were inversely related to victims' educational level. Children under the age of 5, women over 55 years old and men over 60 years old with an educational level lower than the primary school are the most risky groups of populations for intervention measures.
Accidental Falls ; mortality ; statistics & numerical data ; Adolescent ; Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; Educational Status ; Female ; Humans ; Male ; Middle Aged ; Sex Distribution ; Young Adult
7.The synergism and mechanism of action of rClone30-hDR5 in combination with TRAIL on HCC.
Tian SUN ; Ze-Shan NIU ; Xue-Ying LIU ; Gui-You TIAN ; Yin BAI ; Fu-Liang BAI ; Jie-Chao YIN ; Dan YU ; Yun-Zhou WU ; De-Shan LI ; Qing-Zhong YU ; Si-Ming LI ; Gui-Ping REN
Acta Pharmaceutica Sinica 2014;49(7):985-992
To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.
Apoptosis
;
Carcinoma, Hepatocellular
;
pathology
;
Caspase 3
;
metabolism
;
Caspase 8
;
metabolism
;
Drug Synergism
;
Hep G2 Cells
;
Humans
;
Liver Neoplasms
;
pathology
;
Real-Time Polymerase Chain Reaction
;
Receptors, TNF-Related Apoptosis-Inducing Ligand
;
pharmacology
;
TNF-Related Apoptosis-Inducing Ligand
;
pharmacology
;
Transfection
8.Antitumor efficacy of the recombinant Newcastle disease virus rNDV-IL15 on melanoma models.
Ze-Shan NIU ; Fu-Liang BAI ; Tian SUN ; Hui TIAN ; Jie-Chao YIN ; Hong-Wei CAO ; Dan YU ; Gui-You TIAN ; Yun-Zhou WU ; De-Shan LI ; Gui-Ping REN
Acta Pharmaceutica Sinica 2014;49(3):310-315
In order to enhance the antitumor efficacy of recombinant Newcastle disease virus, rNDV-IL15 was rescued in this study. Recombinant plasmid prNDV-IL15 was constructed, and BHK21 cells were transfected with the recombinant plasmid. Finally, the recombinant Newcastle disease virus rNDV-IL15 was successfully rescued. The growth curves of these two recombinant viruses were determined. Murine melanoma B16F10 cells were infected with rNDV-IL15 at MOI of 0.1, and the expression level of IL15 in the supernatant was detected by ELISA. The antitumor efficacy of rNDV-IL15 and rNDV was compared in vitro and in vivo. Results showed that prNDV-IL15 was constructed and recombinant virus rNDV-IL15 was successfully rescued. The growth curve of rNDV-IL15 showed that the growth of rNDV-IL15 had not been changed after insertion of IL15 gene. Results showed that there was high level of IL15 expression in the supernatant of rNDV-IL5-infected B16F10 cells (1 044.3 +/- 27.7 ng x mL(-1)). rNDV-IL15 and rNDV significantly inhibited the growth of B16F10 cells in vitro in a time-dependent manner. However, there was no significant difference between them. In animal experiments, rNDV-IL15 efficiently suppressed tumor growth in vivo when compared with rNDV, and the difference was statistically significant. The results suggested that rNDV-IL15 is a more effective antitumor agent.
Animals
;
Body Weight
;
Cell Line, Tumor
;
Cell Proliferation
;
Chick Embryo
;
Cytotoxicity, Immunologic
;
Female
;
Genetic Therapy
;
Interleukin-15
;
genetics
;
metabolism
;
Melanoma, Experimental
;
pathology
;
therapy
;
Mice
;
Neoplasm Transplantation
;
Newcastle disease virus
;
genetics
;
Plasmids
;
Recombinant Proteins
;
genetics
;
metabolism
;
Transfection
;
Tumor Burden
9.Preliminary biomarker related to nasopharyngeal carcinoma filtered from the whole genome expression profiling involved in microdissection nasopharyngeal tissues.
Zhong-qi LIU ; Yong-quan TIAN ; He HUANG ; Hou-de ZHOU ; Qiu-hong ZHANG ; Ming ZHOU ; Cong PENG ; Xiao-ling LI ; Gui-yuan LI
Journal of Central South University(Medical Sciences) 2005;30(1):1-6
OBJECTIVE:
To filter biomarkers of nasopharyngeal carcinoma (NPC) by constructing the homogenesis tissue gene expression profiling with the whole human genome GeneChip.
METHODS:
The epithelium cells of the homogenesis NPC and the pure nasopharyngeal normal tissues microdissected from nasopharyngeal biopsy which was preserved in the RNAlater were used to isolate RNA and then to harvest the aRNA through in vitro transcription, and aRNA prober was labled to hybridize to HG-U133. plus 2.0, so the expression profiling of each homogenesis tissue could be constructed.
RESULTS:
Some candidate biomarker genes related to the tumorigenesis of NPC had been filtered by comparing the expression profiling of NPC samples with the expression profiling of normal nasopharyngeal epithelia samples. Any genes regarding the metastasis of NPC might have been selected by comparing the expression profiling of no-metastasis samples with those of the metastasis samples.
CONCLUSION
Using the whole genome GeneChip to construct the expression profiling for the microdissected homogenesis tissue is effective to filter the candidate biomarker genes.
Adult
;
Aged
;
Biomarkers, Tumor
;
Female
;
Gene Expression Profiling
;
Gene Expression Regulation, Neoplastic
;
Humans
;
Male
;
Microdissection
;
Middle Aged
;
Nasopharyngeal Neoplasms
;
genetics
;
Nasopharynx
;
metabolism
;
Neoplasm Proteins
;
biosynthesis
;
genetics
;
Oligonucleotide Array Sequence Analysis
;
Tumor Suppressor Proteins
;
biosynthesis
;
genetics
10.Effect of growth inhibition of the secretory protein SPLUNC1 on Pseudomonas aeruginosa.
Hou-de ZHOU ; Ming-hua WU ; Lei SHI ; Ming ZHOU ; Yi-xin YANG ; Jin ZHAO ; Tan DENG ; Xiao-ling LI ; Shou-rong SHENG ; Gui-yuan LI
Journal of Central South University(Medical Sciences) 2006;31(4):464-469
OBJECTIVE:
To express the recombinant SPLUNC1 protein in HNE1 cells and to study its function of bactericidal and binding to lipopolysaccharide (LPS).
METHODS:
Full length of SPLUNC1 gene was cloned into pCMV-tag4A vector and stably transfected into HNE1 cell lines, the supernatant of cell cultures was collected. After being treated with the supernatant, the Pseudomonas aeruginosa was seeded to LB soft agar plate, and the bacteria clones were counted and analyzed. For in vitro LPS binding assay, LPS was coated to 96-well plates. We incubated in the plate with SPLUNC1 protein, and detected the binded SPLUNC1 protein by ELISA. Incubating the FITC-LPS with the SPLUNC1 stably transfected or control cells, the intracellular intensity of fluorescence was observed under the fluorescence microscope.
RESULTS:
SPLUNC1 inhibited the bacteria clone formation obviously. Although the binding efficiency of LPS and SPLUNC1 in vitro was very low, more FITC-LPS entered into the SPLUNC1 stably transfected cells.
CONCLUSION
SPLUNC1 can inhibit the growth of Pseudomonas aeruginosa and bind LPS, and play an important defensive role in innate immunity of the upper airway.
Cell Line, Tumor
;
Glycoproteins
;
isolation & purification
;
pharmacology
;
Humans
;
Membrane Proteins
;
chemistry
;
Nasopharyngeal Neoplasms
;
genetics
;
pathology
;
Phosphoproteins
;
isolation & purification
;
pharmacology
;
Pseudomonas aeruginosa
;
drug effects
;
Respiratory Mucosa
;
chemistry
;
immunology
;
Respiratory System
;
chemistry
;
immunology
;
Transfection