2.Roles of ROS and TGF-?1 in aldosterone-induced production of PAI-1
Jun YUAN ; Ru-Han JIA ; Yan BAO ; Guo-Hua DING ;
Chinese Journal of Nephrology 2005;0(12):-
Objective To explore the roles of reactive oxygen species(ROS) and TGF-?1 in aldosterone-induced PAI-1 production.Methods Quiescent rat mesangial cells (MCs) were treated by aldosterone.The level of ROS in MCs induced by aldosterone was measured by confecal laser scanning microscopy and the TGF-?1 activity in the supematant of culture was measured by mink lung epithelial cell (Mvllu) proliferation inhibition MTT assay.Then,before the addition of aldosterone,MCs were pretreated with NAC or TGF-?1 neutralizing antibody to decrease cellular ROS or inhibit activity of TGF-?1 induced by aldosterone respectively.PAI-1 mRNA was examined by semi-quantification RT-PCR and PAI-1 protein by Western blotting.Results The intracellular ROS induced by aldosterone increased by 5-fold compared to that of control group,and the activity of TGF-?1 stimulated by aldosterone increased markedly.TGF-?1 neutralizing antibody and NAC effectively decreased aldosterone-induced PAI-1 mRNA expression by 30% and 32%,and PAI-1 protein expression by 21% and 11%,respectively.However,neither TGF-?1 neutralizing antibody nor NAC alone could regulate aldosterone-induced PAI-1 mRNA and protein expression to normal level in 24 hours.Conclusions ROS and TGF-?1 play important roles in up-regulation of aldosterone- induced PAI-1 in MCs.ROS and TGF-?1 are not the exclusive pathway of PAI-1 expression induced by aldosterone in MCs.
3.Screening of differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism
Ru-jia, XIE ; Bing, HAN ; Xin-hua, LUO ; Jun, WU ; Ming-liang, CHENG ; Qin, YANG
Chinese Journal of Endemiology 2010;29(6):604-607
Objective To identify the differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism. Methods Six serum samples were collected from patients with liver injury resulting from coal-burning type of arsenism and healthy subjects(control gruop) in endemic arsenism area. Twodimensional gel electrophoresis(2-DE) was performed to separate serum proteins, after silver staining, the differential expression of proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE map of serum protein patterns of patients and normal control were established successfully. The results showed that there were an average of (824 ± 31 ) spots and (782 ± 42) spots on 2-DE matching of the patients and control groups and the matching rate was 94.9%(782/824). From these two groups 49 differential protein spots were identified, of which over 3 times the difference in the expression of 30 protein spots were singled out and MALDI-TOF-MS analysis was carried out. Ten proteins were identified. Upregulated expression was observed in alpha-2-macroglobulin, B-cell receptor-associated protein, keratin 1,apolipoprotein A-I, and down-regulated expression was observed in haptoglobin, α2-heremans-schimid-glycoprotein,mitogen-activated protein kinase 4, zinc finger protein 323, ZAP-70 and SP40 in the patient group. Conclusions The well-resolved and reproducible 2-DE serum patterns of patients are established and some differentially expressed proteins are characterized. Whether these proteins of differential expression are serum markers for liver injury resulting from coal-burning type of arsenism need to be further verified.
4.Bladder saline perfusion before catheter removal in TURP patients
Ru CHENG ; Hui WANG ; Li LI ; Lei HEI ; Shuqian JIA ; Ling HAN ; Yong XU
Chinese Journal of Urology 2012;33(6):448-450
Objective To evaluate the clinical significance of bladder saline perfusion before catheter removal in TURP patients. Methods From 2009 to 2011,140 patients received TURP were enrolled in this study.Patients were divided into perfusion group (70 cases with bladder saline perfusion before catheter removal) and control group (70 cases without perfusion). Results Comparing with the control group (33.1 ± 5.4) min,the time waiting for urination was shorter in perfusion group ( 3.7 ± 0.2 ) min ( P <0.05 ).The recovering time to normal urination was shorter in perfusion group (7.7 ± 1.2 ) d than in control group (11.7 ± 1.3) d (P < 0.05 ) as well.In the first urine after catheter removal and first urine on the next day morning,white blood cell count of 2 groups (4.5 ± 0.1 ) vs ( 6.9 ± 3.5 ) ; ( 3.7 ± 0.2 ) vs (4.3 ±0.5) had significant differences ( P < 0.05 ). Conclusion Bladder saline perfusion before catheter removal in TURP patients is simple and effective for the restoration of normal voiding.
5.Impact of Antimicrobial Agents on Transcription of Gene hld in Forming Process of Staphylococcus epidermidis Biofilm
Qian-Guo HAN ; Xue-Zhong LEI ; Lei ZHANG ; Xiao-Ju LV ; Ru-Jia YU ;
Chinese Journal of Nosocomiology 2009;0(16):-
OBJECTIVE To investigate the transcription level of gene hld of Staphylococcus epidermidis in the biofilm forming and detaching under MIC antibiotic and explore the relationship between biofilm-related drug resistance and persistant infection.METHODS The transcription level of gene hld of S.epidermidis under MIC concentration of 4 antibiotics was compared with those of the control group without antibiotics by SYBR real-time fluorescent quantitative RT-PCR at the different time point of biofilm formation and detachment.RESULTS The transcription of gene hld decreased rapidly from initial adherence,and droped continuously for few hours.There was an increase from 24 hours to 72 hours in groups without antibiotics but not in antibiotics groups,the differenet was significant.CONCLUSIONS Antibiotics improve adherence at first and then prevent matrix decomposition water-conducting tube and detachment of cells by impact of transcription of gene hld,it can protect cells from killing by inhibiting the penetration of biotics and prevent them become planktonic cells after detachment from biofilm.
6.Extraction of sperm DNA from mixed stain by the modified differential lysis method combined with silicon bead method.
Hai-Jun HAN ; Yu-Hong ZHANG ; Min YANG ; Hai YI ; Geng-Ye YANG ; Dong-Tao JIA ; Da-Ru LU
Journal of Forensic Medicine 2014;30(1):50-54
OBJECTIVE:
To extract sperm DNA from mixed stain by the modified differential lysis method combined with silicon bead method and to evaluate its application value.
METHODS:
Fifty-two mixed stains containing female STR genotypes detected by differential lysis method were collected. The sperm DNA was extracted by the modified method combined with silicon bead method, then genotyped with the Identifiler Kit, and compared with the results of genotyping by the conventional differential lysis method as control.
RESULTS:
Of the 52 samples, 38 samples with sole male STR genotypes in all loci were detected. The detection rate of male STR genotypes was 98.08% through the modified method combined with silicon bead method.
CONCLUSION
The modified differential lysis method combined with silicon bead method can be used in extraction of sperm DNA from mixed stain.
DNA/isolation & purification*
;
DNA Fingerprinting
;
Female
;
Genotype
;
Humans
;
Male
;
Polymerase Chain Reaction
;
Silicon
;
Spermatozoa
7.Progress on Correlation between the Expression of CDK5 and Brain Injury Time.
Shi-yu MA ; Ru-bo LI ; LUO YU-JIA ; Meng-yan LÜ ; Han-zhi WANG ; Zheng-yin WANG
Journal of Forensic Medicine 2016;32(1):58-60
Cyclin-dependent kinase 5 (CDK5) is a member of cyclin-dependent kinase family, which does not directly regulate cell cycle. Through phosphorylation of target protein, CDK5 plays an irreplaceable role in the development, reparation and degeneration of neurons. Brain injury refers to the organic injury of brain tissue caused by external force hit on the head. Owing to the stress and repair system activated by our body itself after injury, various proteins and enzymes of the brain tissues are changed quantitatively, which can be used as indicators for estimating the time of injury. This review summarizes the progress on the distribution, the activity mechanism and the physiological effects of CDK5 after brain injury and its corresponding potential served as a marker for brain injury determination.
Brain/physiopathology*
;
Brain Injuries/physiopathology*
;
Cyclin-Dependent Kinase 5/metabolism*
;
Nerve Tissue Proteins/metabolism*
;
Neurons
;
Neuroprotective Agents/pharmacology*
;
Phosphorylation/drug effects*
;
Time Factors
8.Changes of bone morphogenetic protein-7 and inhibitory Smad expression in streptozotocin-induced diabetic nephropathy rat kidney.
Qin YANG ; Bing HAN ; Ru-Jia XIE ; Ming-Liang CHENG
Acta Physiologica Sinica 2007;59(2):190-196
The present study was designed to observe the expressions of bone morphogenetic protein-7 (BMP-7) and inhibitory Smads in kidney of rats with diabetic nephropathy (DN), and explore the possible mechanism of DN. Male Wistar rats weighing 180-220 g were single injected with streptozocin (STZ, 55 mg/kg body weight) for 2, 4, 8 and 16 weeks to induce DN. Blood glucose, kidney weight/body weight and 24-hour urine protein in the control and DN rats were examined; the expressions of BMP-7, Smad6 and Smad7 were detected by using immunohistochemical techniques, Western blot and real-time PCR. The results showed that blood glucose and 24-hour urine protein in DN rats were higher than that in the control rats and kidney weight/body weight was also elevated in DN rats, especially in 16-week STZ-induced rats. The expressions of BMP-7 and Smad6 proteins in DN rats were elevated, while BMP-7 mRNA expression was increased 2 weeks after STZ injection and decreased 16 weeks after STZ injection. The expressions of Smad7 protein and mRNA were elevated in DN rats 2 weeks after STZ injection and decreased 16 weeks after STZ injection. In addition, the expressions of transforming growth factor-beta1 (TGF-beta1) and collagen type I (COL-I) mRNA were increased in DN rats. These results suggest in the early stage of DN, increase in BMP-7 and inhibitory Smad expression may play a role in the feedback regulation and restrain the development of DN.
Animals
;
Bone Morphogenetic Protein 7
;
genetics
;
metabolism
;
Collagen Type I
;
metabolism
;
Diabetes Mellitus, Experimental
;
metabolism
;
Diabetic Nephropathies
;
metabolism
;
Kidney
;
metabolism
;
pathology
;
Male
;
RNA, Messenger
;
genetics
;
metabolism
;
Random Allocation
;
Rats
;
Rats, Wistar
;
Smad6 Protein
;
genetics
;
metabolism
;
Smad7 Protein
;
genetics
;
metabolism
;
Transforming Growth Factor beta1
;
metabolism
9.Advances in targets of pharmacotherapy and discovery of new drugs for Alzheimer disease:do we have any novel strategies?
wei Wei HU ; qin Xue HOU ; jia Ru LIAO ; ting Han ZHANG
Chinese Journal of Pharmacology and Toxicology 2017;31(7):701-706
Alzheimer disease (AD), the most common dementia, is a chronic, progressive and neuro-degenerative disorder. With an increasing prevalence, AD has been the third cause of death after cardio-vascular diseases and cancer in the elderly population. However, the pathogenesis of AD remains unclear, which has led to a fairly slow development of drugs for AD and a dim view of future treatments of AD. It has been a hot spot and a big challenge to develop effective, therapeutic drugs for AD. Recently, this topic was discussed via WeChat by experts from the Neuropsychiatric WeChat Group, which consists of 300 Chinese-origin neuroscientists and neuropsychiatrists in China or overseas. The experts pointed out the problems that might have misled researches on drug discovery, such as the misleading but dominating AD pathology hypotheses and problems with the platforms for drug screening. Therefore, it is important to review the pathology of AD and the treatment strategies from big data and the overall view of the disease, which may shed new light on AD therapy to develop drugs for multiple targets, leading to omni-direc-tional, comprehensive treatments of AD. The development of AD can be further classified into different stages based on the upstream factors of AD pathology. Interestingly, it has been found that the AD brain has mitochondria damage and dysfunction; long-term exposure to low doses of ionizing radiation can also cause AD-like pathological changes. These provide novel views and ideas in terms of the path-ological process and preventive and therapeutic strategies for AD.
10.Dysbiosis of lung commensal bacteria in the process of lung epithelial-mesenchymal transition in mice with silicosis
China Occupational Medicine 2022;49(05):514-
Objective -
To investigate the effect of lung flora dysbiosis on the process of pulmonary fibrosis and lung epithelial
( ) Methods -
mesenchymal transition EMT in mice with silicosis. Male C57BL/6 mice of specific pathogen free grade were
, , , ( )
randomly divided into the blank control group silicosis model group solvent control group vancomycin VM + ampicillin
( ) , ( ) ( ) ,
AMP group metronidazole MNZ + neomycin NEO group and mixed treatment group 12 mice in each group. Except for
, ,
the blank control group which was given 20.0 µL of 0.9% NaCl solution the other five groups of mice were dosed with 20.0 µL
of silica dust suspension at a mass concentration of 250.0 g/L using a single tracheal drip to establish the silicosis mouse model.
:
The intranasal drip method was used to treat silicosis mice in each group as following mice in the solvent control group were
- ; ;
given double distilled water mice in the VM+AMP group were given VM at a mass concentration of 0.5 g/L and AMP at 1.0 g/L
;
mice in the MNZ+NEO group were given MNZ at a mass concentration of 1.0 g/L and NEO at 1.0 g/L mice in the mixed
,
treatment group were given the same doses of the four antibiotics mentioned above all in a drip volume of 50.0 µL. Silicosis
, ,
mice were treated seven days and half an hour before silica dusting and 7 14 and 21 days after silica dusting. Mouse lungtissue was collected aseptically 28 days after silica dusting. Hematoxylin eosin and Masson trichrome staining methods were
-
used to observe the pathological changes. Western blotting was used to detect the relative protein expression of α smooth muscle
( - ), - ( - ) ( )
actin α SMA E cadherin E CAD and vimentin VIM . Immunohistochemistry was used to detect the relative expression of
- -
E CAD and VIM. Real time fluorescence quantitative polymerase chain reaction was used to detect the expression levels of
(Col1a2) Results
collagen type Ⅰ alpha 2 mRNA in lung tissues. The histopathological results showed that the alveoli of the
,
blank control group were thin and structurally intact with few surrounding infiltrating inflammatory cells and no abnormal
,
distribution of collagen fibers. The alveoli of the silicosis model group were structurally disorganized with a large number of
, ,
infiltrating inflammatory cells thickened alveolar walls and cellular fibrous nodules with abundant blue collagen deposit. In the
, ,
VM+AMP group MNZ+NEO group and the mixed treatment group the inflammation and fibrosis were reduced with diferent
degrees in the lung tissues compared to the silicosis model group and the solvent control group. The relative expression levels of
- , Col1a2
α SMA VIM protein and mRNA in lung tissues of mice in the silicosis model group were higher than those in the blank
( P ), -CAD
control group all <0.05 and the relative expression levels of E protein were lower than those in the blank control
(P ) - , Col1a2
group <0.05 . The relative expression levels of α SMA VIM protein and mRNA in lung tissues of mice in the MNZ+
( P ), -CAD
NEO group and the mixed treatment group were lower all <0.05 and the relative expression levels of E protein were
(P ), Conclusion
higher <0.05 when compared with the silicosis model group and the solvent control group. Pulmonary fibrosis
, -
was reduced in silicosis mice with interventions in lung flora where anaerobic and gram negative bacteria affected pulmonary
fibrosis and dysbiosis of the lung flora affected pulmonary EMT.