1.Effects of moxibustion pretreatment on extracellular signal-regulated kinase signaling transduction pathway in the gastric tissues of rats with gastric mucosal damage
Wenna SHU ; Qing YANG ; Huan ZHONG ; Mi LIU ; Sian PAN ; Lina GUO ; Zhao ZHAO ; Chao WANG ; Xiaorong CHANG
Journal of Acupuncture and Tuina Science 2016;14(3):149-155
Objective: To observe the effects of moxibustion pretreatment on the protein expressions of epidermal growth factor receptor (EGFR), phosphorylation extracellular signal-regulated kinase 1/2 (p-ERK1/2) and activated protein-1 (AP-1), the key factors of extracellular signal-regulated kinase signaling transduction pathway in gastric tissue of rats with stress-induced gastric mucosal damage, and to discuss the mechanisms of moxibustion therapy in promoting the restoration of damaged gastric mucosa.
Methods: Thirty Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, and a moxibustion group using the random digits table, 10 in each group. Except the rats in the normal group, rats in the other two groups were used to make stress-induced gastric mucosal damage model using restraint and cold stress. Before modeling, rats in the moxibustion group were alternately treated with moxibustion at Zusanli (ST 36) and Zhongwan (CV 12), or Pishu (BL 20) and Weishu (BL 21), once a day, for a total of 8 d. Histological changes of gastric mucosa were observed under the light microscopy, the expression of gastric tissue p-ERK1/2 was detected by immunohistochemistry assay, and the protein levels of EGFR and AP-1 were measured by Western blots.
Results: Compared with rats in the normal group, gastric mucosal damage was more serious, and protein expressions of gastric tissue EGFR, p-ERK1/2 and AP-1 increased in the model group (P<0.01,P<0.05,P<0.05). Compared with rats in the model group, gastric mucosal damage was milder, and protein expressions of gastric tissue EGFR, p-ERK1/2 and AP-1 increased in the moxibustion group (allP<0.01).
Conclusion:Moxibustion at Zusanli (ST 36), Zhongwan (CV 12), Pishu (BL 20) and Weishu (BL 21) couldincrease EGFR, p-ERK1/2 and AP-1 expression levels in gastric tissue of stress-induced gastric mucosal damage rats, maintain the information transfer function of ERK signaling transduction pathway, and promote restoration of gastric mucosal damage.
3.The development and clinical application of papillomavirus genotyping by DNA chip.
Guang YANG ; Cai-hong LIANG ; Jin-huan CUI ; Shu CHEN
Chinese Journal of Epidemiology 2006;27(1):47-49
OBJECTIVETo develop a new platform for genotyping human papillomavirus (HPV) and to investigate its effect in clinical application.
METHODSA pair of common primers of 18 HPV subtypes for PCR, was designed in HPV conservative L1 region. Genotyping probes for detecting 15 high-risk HPV subtypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66 and 68, together with 3 low-risk HPV 6, 11 and 42 were selected respectively from Genbank and fixed on membrane to make DNA chip. PCR amplification and DNA chip technology were optimized. 100 clinical samples were used to investigate the effect of HPV genotyping DNA chip. Veracity of the genotyping results was verified by sequencing.
RESULTSFrom the 100 clinical samples, 30 were found to be HPV positive, including high-risk HPV subtypes 16, 18, 33, 45, 51, 58, and 66, and low-risk HPV 6, 11 and 42. The sensitivity tested by standard samples was up to 10 copies of HPV DNA.
CONCLUSIONThe HPV genotyping system developed here with DNA chip showed high sensitivity and specificity, suitable to be applied in clinical practice for HPV diagnosis and investigation on the prevalence of HPV sub-types.
DNA Probes, HPV ; genetics ; DNA, Viral ; genetics ; Female ; Genotype ; Humans ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; methods ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; virology ; Polymerase Chain Reaction ; Sensitivity and Specificity ; Uterine Cervical Neoplasms ; virology
4.Clinical diagnosis,treatment and outcome of cerebral vein thrombosis:analysis of 11 cases
Juan BAO ; Xiao-Shu YANG ; Xiao-Yan LONG ; Jian XIA ; Huan YANG ; Jing LI ; Qi-Dong YANG
Chinese Journal of Neuromedicine 2009;8(9):923-925
Objective To analyze the clinical manifestations,radiographic characteristics and treatment of cerebral vein thrombosis.Methods The clinical manifestations,results of laboratory examinations,characteristic radiographic findings,treatment protocols and outcomes were analyzed retrospectively in 11 patients with cerebral vein thrombosis admitted between 2002 and 2007.Results In 10 of the patients,nonspecifie headache was the most frequent symptoms,followed by vomiting,hemiplegia,meningeal irritation and hyperspasmia.Two patients received treatments for suspected cerebral hemorrhage and subarachnoid hemorrhage before a definite diagnosis was established.All thel 1 patients received CT and/or magnetic resonance imaging(MR/)examinations,and 8 also underwent magnetic resonance venography(MRV)and 1 underwent digital subtraction angiography(DSA),and a definite diagnosis of cerebral vein thrombosis was established in 10 eases.Treatment to control the intracmnial pressure was administered in all the patients,among whom 10 were given anti-coagulation or anti-platelet treatments.Nine patients showed improvements after the treatments;1 had deteriorated condition and 1 died.Conclusion In the absence of specific clinical manifestations,cerebral vein thrombosis gives rise to high rate of misdiagnosis,and a definite diagnosis relies on the findings by radiographic modalities.Early anti-coagulation treatment may prove safe and effective in these cases.
5.Comparison of data population-based and from hospital-based injuries.
Ling-ni ZHOU ; Jie-min MA ; Zhong-jie LI ; Shu-yang CHEN ; Gong-huan YANG
Chinese Journal of Epidemiology 2004;25(11):967-969
OBJECTIVETo compare data from an epidemiological survey on injuries with a survey conducted in hospitals on injuries in the same areas and to find out the differences and shortcomings of hospital data in describing the feature of injuries in an area.
METHODSComparing the causes and age distributions of injuries from the two surveys.
RESULTSThe first 4 leading causes of injuries from the population-based survey were mechanical injuries, falls, burns/scalds and traffic accidents while the first 4 leading causes of hospital-based survey were traffic accidents, assault, mechanical injuries and burns/scalds. The differences of the age distributions of these leading causes between the two surveys were significant except mechanical injuries.
CONCLUSIONDifferences were noticed between population-based survey and hospital-based survey. It should be cautions when using hospital data to describe the features of injuries in a certain area.
Accidental Falls ; statistics & numerical data ; Accidents, Traffic ; statistics & numerical data ; Adolescent ; Adult ; Burns ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Hospitalization ; Humans ; Incidence ; Male ; Middle Aged ; Rural Health ; Wounds and Injuries ; epidemiology
6.Development and application of a new hepatitis C virus genotyping method with polymerase chain reaction-reverse blot dot technique.
Guang YANG ; Shu CHEN ; Jin-huan CUI ; Jian-hua SI ; Jia-ju TAN
Chinese Journal of Epidemiology 2005;26(6):440-443
OBJECTIVEUsing polymerase chain reaction-reverse blot dot (PCR-RDB) technique to establish a new method for hepatitis C virus (HCV) genotyping and to study the distribution of HCV genotypes in Foshan area.
METHODSHCV primers and probes were designed in 5'-untranslated region (nt-1-nt-299) of HCV. HCV RNA in serum was isolated and purified, and its cDNA was obtained by reversed transcription. Nested PCR using biotin-labelled primers, was done. PCR products were hybridized with immobilized specific probes (genotype 1a to 3b) on Biodyne C membrane to genotype HCV by color development while adding POD and TMB. A certain judgment could be made according to the position of color reaction. The reliability of this new method was verified by sequencing. HCV RNA levels in serum were determined by real time fluorescent quantitative (FQ)-PCR. 60 FQ-PCR-positive HCV sera from Foshan area were genotyped using this assay.
RESULTSAll 60 sera could be successfully genotyped by PCR-RBD. 50 (83.3%) cases were found to be genotype 1b, 2 (3.3%) as genotype 1a and 2 (3.3%) as genotype 2a while 5 (8.0%) to be mixture of genotype 1a and 1b, and 1 (1.7%) to be mixture of genotypes 1b and 2a. No genotypes 2b, 3a and 3b were found. The results of PCR-RDB genotyping methods coincided with sequence analysis.
CONCLUSIONNewly established HCV genotyping system was proved to be sensitive, specific, precise and economic, thus suitable for clinical and epidemiologic studies. The results of HCV genotyping showed that genotype 1b was the predominant genotype in Foshan area.
Genotype ; Hepacivirus ; classification ; genetics ; Hepatitis C ; virology ; Humans ; Immunoblotting ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity
7.Quantification of Tannerella forsythensis in chronic periodontitis patients.
He YANG ; Shu MENG ; Lei ZHAO ; Yi XU ; Ya-fei WU ; Huan ZHAO
West China Journal of Stomatology 2007;25(5):454-457
OBJECTIVETo compare the number of the Tannerella forsythensis (T. forsythensis), total bacteria, and proportion of T. forsythensis in subgingival specimens in diseased sites of chronic periodontitis patients and in healthy sites of periodontally healthy subjects, and clarify the relationship between bacterial load and periodontal status.
METHODSSubgingival plaque samples from 61 chronic periodontitis patients and 12 healthy controls (positive for T. forsythensis by conventional PCR) were analyzed with TaqMan real-time polymerase chain reaction assay for T. forsythensis and total bacteria. Quantification was performed with species-specific primer/probe, universal primer/ probe and serial dilution of plasmid standards.
RESULTSNumbers of T. forsythensis and total bacteria(P<0.001) , the proportion of T. forsythensis in subgingival specimens (P<0.05) were significant higher in diseased sites of chronic periodontitis patients than in healthy sites of healthy subjects. In addition, a significant correlation was found between the number of bacteria and various probing depth (P<0.001). There was no significantly difference between the proportion of T. forsythensis in subgingival plaque and probing depth.
CONCLUSIONNumber of T. forsythensis are closely associated with periodontal status, and demonstrate the broad potential of real-time polymerase chain reaction application on periodontology.
Adult ; Bacteroides ; Chronic Periodontitis ; Dental Plaque ; Female ; Humans ; Male ; Middle Aged ; Periodontitis ; Polymerase Chain Reaction ; Porphyromonas gingivalis
8.Adenoviruses mediated BCL-X1 overexpression protects mice from fulminant hepatic failure.
Xiao-an YANG ; Ka ZHANG ; Xin SHU ; Hong CAO ; Qi-huan XU
Chinese Journal of Experimental and Clinical Virology 2011;25(2):109-111
OBJECTIVETo indentify the relation between hepatic cells apoptosis and the lesion of liver tissue in acute toxic lethal hepatitis.
METHODS60 Wistar mice were randomly divided into normal control, model group and treatment group. Normal control and model group were pretreated by portal vein injection of normal saline, the treatment group was pretreated by portal vein injection of BCL-X1 adenoviruses. The mice of model group and treatment group were received an injection of D-galn and LPS to establish fulminant hepatic failure models 7 days after pretrement. To observe BCL-X1 expression, serum ALT, AST, hepatocyte apoptosis rate, and mortality rate of the three groups.
RESULTSThe BCL-X1 expression was higher in treatment group than in model group; 6 hours after fulminant hepatic failure models were established,the serum ALT, AST level of treatment group was lower than model group;The hepatocyte apoptosis rate of treatment group was lower than model group. The death rate of treatment group was lower than model group.
CONCLUSIONIn fulminant mice hepatic failure models, the hepatocyte apoptosis rate has a positive correlation with death rate, the overexpression of BCL-X1 can decrease the hepatocyte apoptosis rate and the death rate.
Adenoviridae ; genetics ; metabolism ; Animals ; Apoptosis ; Disease Models, Animal ; Female ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; genetics ; metabolism ; Humans ; Liver ; cytology ; metabolism ; Liver Failure, Acute ; genetics ; physiopathology ; prevention & control ; therapy ; Rats ; Rats, Wistar ; bcl-X Protein ; genetics ; metabolism ; therapeutic use
9.Study of 161 chronic idiopathic myelofibrosis patients for clinicopathological staging.
Ai-ning LIU ; Hui-shu CHEN ; En-bin LIU ; Li-huan FANG ; Qing-ying YANG ; Lin-sheng QIAN
Chinese Journal of Hematology 2006;27(3):178-182
OBJECTIVETo investigate the significance of clinicopathological stage of chronic idiopathic myelofibrosis (CIMF) in WHO classification of 2001.
METHODSHistopathological analysis of bone marrow biopsy plastic-embedded sections stained with H-G-E and Gomori's stains and clinical features of 113 cases previously diagnosed as primary myelofibrosis (PMF) and 48 cases MPD-U (total of 161 cases which including male 79 and female 82) were studied retrospectively.
RESULTSThere was no significant differences on the clinical features among the cellular phase, collagen fiber phase, sclerotic phase and osteomyelosclerosis of 113 previously diagnosed patients. According to WHO classification 2001 of CIMF, previously diagnosis in 48 cases with MPD-U was WHO pre-CIMF, and in 113 cases with PMF was WHO CIMF-Fs. There were significant differences between of WHO pre-CIMF and WHO CIMF-Fs about clinicopathological features except age. The percentage of immature granulocytes, normoblasts, lymphocytes in peripheral blood, the size of hepatosplenomegaly, and the percent age of tear drop-like red blood cells in pre-CIMF were significantly lower than those in CIMF-Fs (P < 0.05). However, the number of hemoglobin and platelets in patients with pre-CIMF were significantly higher than that with CIMF-Fs (P < 0.01).
CONCLUSIONpre-CIMF and CIMF-Fs in clinical and histopathological features were different development stage of CIMF, while osteomyelosclerosis is a variant of CIMF, but not an independent disease.
Adult ; Aged ; Biopsy ; Bone Marrow ; pathology ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Primary Myelofibrosis ; classification ; pathology ; Thrombopoiesis
10.Inhibition of osteosarcoma cell proliferation by a short hairpin RNA targeting proliferation cell nuclear antigen.
Qi-liang ZHANG ; Shu-hua YANG ; Hong-yun LIU ; Cui-huan WU
Chinese Journal of Pathology 2005;34(3):167-170
OBJECTIVETo construct a plasmid vector expressing the short hairpin RNA (shRNA) targeting proliferation cell nuclear antigen (PCNA), and to investigate its effect in vitro on the expression of PCNA, proliferation and apoptosis of human osteosarcoma cells.
METHODSA plasmid vector expressing the short hairpin RNA targeting at PCNA was constructed and transfected into human osteosarcoma cell line MG-63 by dosper liposomal method. PCNA mRNA and protein expressions were examined using RT-PCR and immunohistochemical staining, respectively. Inhibition of the cell proliferation was studied by MTT method and colony forming assay. DNA synthesis was analyzed by (3)H-TdR incorporation and the cell cycle was determined by flow cytometry. The apoptotic cells were stained with acridine orange.
RESULTSExpression of PCNA mRNA after the transfection was markedly inhibited by 80.51% with a PI value of 25.68% of that of the control group. PCNA shRNA inhibited MG-63 cell growth detected by using MTT method, with an inhibition rate of 61.78% at 48 h. DNA synthesis rate also decreased in the (3)H-TdR incorporation test. Flow cytometry analysis showed an increase of the percentage of G(0)/G(1) phase cells, along with a decrease of cell population in the S phase. The apoptosis rate of cells transfected with the plasmid vector was 16.54%.
CONCLUSIONSPCNA shRNA significantly suppresses the expression of PCNA at both mRNA and protein levels, corresponding to an inhibition of the proliferation of MG-63 cell and an increase of the cellular apoptosis.
Apoptosis ; Bone Neoplasms ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Osteosarcoma ; metabolism ; pathology ; Plasmids ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; biosynthesis ; genetics ; Transfection