1.The transcription factor Egr-1 and the lung diseases
Ling CHU ; Qing-Fu ZENG ;
Chinese Journal of Pathophysiology 1986;0(02):-
Egr-1 is an important transcription factor, which regulates at least 30 kinds of gene expression. Egr-1 couples extracellular signals to long-term responses by altering expression of Egr-1 target genes. So egr-1 can directly or indirectly affect cell differentiation,apoptosis,immune response,injury and repair. This article reviewed the progress in Egr-1 and the lung disease.
2.Construction of Human Papillomavirus Type 18 E6E7 Antisense RNA Expressing Recombinants
Qing GUO ; Fanqin ZENG ; Ling LU
Chinese Journal of Dermatology 1995;0(03):-
Objective In order to study the pathogenesis of human papillomavirus(HPV) and seek for a therapeutic approach of the diseases caused by HPV, the construction of HPV18 E6E7 antisense RNA expressing recombinants was studied. Methods We amplified the HPV18 E6E7 816bp by PCR with HPV18 plasmid DNA as the template. pLNSX retroviruses were used as vectors,the HPV18 E6E7 retrovirus recombinants were constructed. And then the recombinants were cleaved with restriction endonuclease and hybridized with Southern blot for identifying the inserting direction and special check respectively. Results and conclusion The HPV18 E6E7 antisense RNA retrovirus expressing recombinants were screened and obtained,which had laid the foundation of studying the function of E6E7 genes further and explore whether the antisense technique can adjust and control the expression of E6E7 genes.
3.Small hairpin loop RNA targeting HIF-1alpha down-regulates VEGF and up-regulates PEDF in human retinal pigment epithelial cells under hypoxic condition.
Qing, XIAO ; Shuiqing, ZENG ; Mingliang, LV ; Shiqi, LING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2008;28(4):460-4
The aim of this study was to explore the effect of small hairpin loop RNA (shRNA) silencing hypoxia-induced factor 1alpha (HIF-1alpha) gene on the expression of vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) in human retinal pigment epithelium (RPE) cells under hypoxic condition. Two target sites of HIF-1alpha mRNA were chosen and two kinds of shRNA were designed and synthesized against the target sites. Then the two kinds of shRNA were transfected into human RPE cells in vitro, respectively. These cells were cultured under hypoxic condition that was simulated by using 150 mumol/L CoCl(2). The mRNA expressions of HIF-1alpha, VEGF and PEDF were tested by semi-quantitative reverse transcription PCR (RT-PCR). The protein levels of HIF-1alpha, VEGF and PEDF were analyzed by Western blotting. After the two kinds of HIF-1alpha-specific shRNA were transfected into RPE cells respectively, the expression of HIF-1alpha mRNA and the levels of HIF-1alpha protein were decreased significantly in RPE cells under hypoxic condition. The expression of VEGF mRNA and the levels of protein significantly were also decreased. However, the levels of PEDF protein was significantly increased, but the expression of PEDF mRNA showed no significant changes. In conclusion, HIF-1alpha-specific shRNA can effectively silence the HIF-1alpha gene, and consequently down-regulate VEGF and up-regulate PEDF expression against hypoxia. These results reveal that HIF-1 is associated with posttranslational mechanism for down-regulating PEDF under hypoxia and provide an explanation for hypoxia-provoked increases in VEGF/PEDF ratios. These results also suggest that HIF-1 is one of the key cytokines to retinal neovascularization.
4.Up-regulation of HIF-1alpha and VEGF expression by elevated glucose concentration and hypoxia in cultured human retinal pigment epithelial cells.
Qing, XIAO ; Shuiqing, ZENG ; Shiqi, LING ; Mingliang, LV
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(4):463-5
In order to explore the effect of high glucose concentration and high glucose concentration with hypoxia on the production of hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF), human RPE cells were cultured in 5.56 mmol/L glucose (control group), 5.56 mmol/L glucose with 150 micro mol/L CoCl2 (hypoxic group), 25 mmol/L glucose (high glucose group) and 25 mmol/L glucose with 150 micro mol/L CoCl2 (combination group). RT-PCR was used to detect the expression of HIF-1alpha and VEGF mRNAs. Western blot analysis was used to measure the levels of HIF-1alpha and VEGF proteins. Although the small amount of HIF-1alpha protein was able to be detected in high glucose group but not in control group, there was no significant difference between the expression of HIF-1alpha mRNA of RPE cells in high glucose group and that of RPE cells in control group. As compared with RPE cells in control group, the mRNA expression and the protein synthesis of VEGF in high glucose group were up-regulated. As compared with RPE cells in hypoxic group, the expression of HIF-1alpha mRNA of RPE cells in combination group was not different, but the protein synthesis of HIF-1alpha, the mRNA expression and the protein synthesis of VEGF were more obviously up-regulated. In conclusion, high concentration glucose mainly influence the protein synthesis of HIF-1alpha of RPE cell, and HIF-1alpha protein is able to be accumulated in high concentration glucose. Under hypoxia, the HIF-1alpha protein induced by high concentration glucose is more stable, and the expression of VEGF is obviously increased. It is suggested that high concentration glucose may play a role in retinal neovascularization, especially at ischemia stage of diabetic retinopathy.
5.Selecting the tag SNPs of TLR2 gene with bioinformatics techniques
Kehong CHEN ; Ling ZENG ; Wei GU ; Shuna HUANG ; Qing LIU ; Jianxin JIANG
Basic & Clinical Medicine 2010;30(3):242-245
Objective To select the tag SNPs of TLR2 gene in the Chinese population with bioinformatics techniques.Methods We ascertained the assayed scope of the TLR2 gene with the aid of NCBI database and downloaded SNP genotype data of TLR2 gene in the Chinese population from Hapmap database.Then Haploview (version 4.0) was used to calculate linkage disequilibrium (LD) statistics.Haplotype blocks were constructed throughout the TLR2 gene according to the upper and lower 95% confidence bound of the D'value.Meanwhile,we selected the tag SNPs based on r~2 values and the LOD value between SNPs and picked up the representative haplotypes in accordance with the proportion of each haplotype in the haplotype blocks,respectively.Results We constructed 2 haplotype blocks within the TLR2 gene and selected 3 tag SNPs containing 3013 A/G,19216 T/C and 22215 G/T in the Chinese population.Meanwhile,we identified the representative haplotypes of which the tag SNP would be on behalf of every haplotype block.Conclusion The SNPs of 3013 A/G,19216 T/C and 22215 G/T,the most representative SNPs in the whole TLR2 gene in the Chinese population,could be selected as tag SNPs to guide their association studies between the TLR2 gene and sepsis.
6.Emergency reparation and reconstruction for complicated tissue defects in hand
Yi-Min CHAI ; Bing-Fang ZENG ; Qing-Ling KANG ; Jian-Feng XUE ; Ji SHEN ; Jia JIANG ;
Chinese Journal of Microsurgery 2006;0(05):-
Objective To explore the operative methods and clinical outcomes in emergency or sube- mergency repair for the complicated tissue defects in hand in first stage applying microsurgical technique. Methods From Jan.,2000 to Aug.,2005,49 emergency cases of complicated hand tissue defects were re- paired in the first stage with replantation,reconstruction,free flaps,combined finger reconstruction and flap transplantation,including 21 cases in mini tissue mass replantation or reconstruction,15 cases in replantation combined with free flap transplantation,8 cases in replantation and reconstruction combined with free flap transplantation,5 cases in combined multiple digits reconstruction with free flap transplantation.The free flap transplantation included the anterolateral femoral flap,the latissimus dorsi myocutaneous flap,the dorsalis pe- dis flap,the media pedis flap and the instep flap.Results All the flaps,the replanted and reconstruceted finger survived uneventfully except for one replanted finger necrosis.45 cases healed in the first stage and the other 4 cases healed in the second stage.During a follow-up from 6 months to 3 years postoperatively,a satis- factory appearance and function of the reconstructed hand was achieved.The excellent and good rate was 85.7% assessed with provisional functional assessment criterion for upper limbs issued by Chinese Society of Hand Surgery.Conclusion The emergency or subemergency repair for the complicated tissue defects in hand has the advantage of short-term treatment and desirable functional outcome.The emergency replantation and reconstruction combined with various flaps or tissue mass can be applied to repair tissue defect in hand in the first stage according to the position and area of the defect along with the technique level of the surgeon, having been proved to achieve desirable clinical outcomes.And the key points leading to a successful operation is the correct treatment for the raw surface of the defects,suitable choice for various flaps,logical design of combination pattern and prevention and timely treatment for vessel crisis.
7.Influence of intravitreal avastin on the expression of cell factors in retinal proliferative membrane in proliferative diabetic retinopathy eye
Bo-jie, HU ; Qing, ZENG ; Xin-ling, LIU ; Xiao-rong, LI ; Wen-jing, SONG
Chinese Journal of Experimental Ophthalmology 2013;(1):55-59
Background A main cause of visual impairment in proliferative diabetic retinopathy (PDR) is vitreous hemorrhage and retinal detachment due to contraction of fibrovascular membrane.To explore the pathogenic mechanism of fibrovascular membrane is a new target for the prevention and management of PDR.Objective This study was to determine the change in expression of vascular endothelial growth factor (VEGF),connective tissue growth factor(CTGF) and pigment epithelium derived factor(PEDF) in the proliferative membranes of patients with PDR after intravitreal injection of avastin,an anti-VEGF agent.Methods This study was approved by the Medical Ethic Committee of Tianjin Medical College,and written informed consent was obtained from each patient before enrollment.A prospective randomized-controlled study was designed.Twenty-six eyes of 24 patients with PDR scheduled for surgery were enrolled from January to June,2008 in Tianjin Medical College Eye Hospital.The patients were randomized into the simple vitrectomy group and avastin injection combined with vitrectomy group,with matched gender,age and disease duration.1.25 mg (0.05 ml) of avastin was intravitreally injected prior to surgery,and vitrectomy was performed 10 days after injection in the avastin injection combined with vitrectomy group,and only vitrectomy was given in the simple vitrectomy group.Preretinal membrane was collected during the surgery.Expression of VEGF,CTGF and PEDF in the preretinal membranes was assayed by immunochemistry.Results VEGF,CTGF and PEDF were expressed in the cytoplasm.The rate of VEGF expression in the preretinal membranes was 30.77% in the avastin injection combined with vitrectomy group,showing a significant reduction in comparison with the simple vitrectomy group(100.00%)(U =4.000,P<0.01).The rate of expression CTGF was remarkable elevated in the avastin injection combined with vitrectomy group compared with the simple vitrectomy group (92.31% vs.62.54%)(U=7.500,P=0.048).However,no significant difference was found in the expression rate of PEDF between the two groups(100.00% vs.92.31%) (U =65.500,P =0.299).Conclusions The results suggest that intravitreal injection of anti-VEGF drugs resulted in the decrease of VEGF expression and increased CTGF expression in proliferative membranes from patients with PDR.
8.MK-801 attenuates NOS expression and NO content in the spinal cord of rats with inflammatory pain
Jing-Bo ZENG ; Wen-Bin LI ; Qing-Jun LI ; Xiao-Ling CHEN ; Ai-Min ZHOU ; Yi-Ling LING
Acta Physiologica Sinica 2001;53(1):55-60
Using NADPH-d histochemistry, the effect of NMDA receptor antagonist MK-801 on the expression of nitric oxide synthase (NOS) in the dorsal horn of the spinal cord was investigated during inflammatory pain and hyperalgesia induced by injection of formalin into the right hind paw. The course of the change in the nitric oxide (NO) content of the lumber intumescence was observed by measuring the ratio of nitrate/nitrite (NO3/NO2) and also the end product of NO. The results showed that the NOS expression and NO contents significantly increased 24 h after formalin injection, which were substantially inhibited when MK-801 was intrathecally injected 15 min prior to formalin injection or 12 h after formalin injection. The results suggest that the increases in the expression of NOS and NO contents in the dorsal horn of the spinal cord are mediated by activation of NMDA receptors during pain and hyperalgesia after formalin injection.
9.The characteristics of the genes mutations in rifampin and isoniazid resistant Mycobacterium tuberculosis clinical isolates from Baise district, Guangxi autonomous region
Hongyu WEI ; Xinying LONG ; Jun LING ; Zhenfeng XIE ; Huaying TANG ; Xiaofeng HUANG ; Liandeng WEI ; Yanchun QING ; Yi ZENG
The Journal of Practical Medicine 2015;(5):731-734
Objective To analyze the characteristics of the rpoB, KatG and inhA genes mutations in rifampin and isoniazid resistant Mycobacterium tuberculosis (MTB) clinical isolates in Baise district, Guangxi autonomous region. Methods 128 MTB clinical strains were collected and isolated for drug susceptibility testing, and drug resistant strain DNA was subtracted for rpoB, KatG and inhA genes mutation analysis. Results 75%(27/36)isolates carried mutations in the rpoB gene,and 59.3%(16/27)isolates carried mutations in 531 sites. 44.1%(15/34) isolates carried mutations in KatG or inhA, and 66.7%(10/15) isolates appeared in KatG 315 site, with two new mutations found in KatG 279 and 427 site. In these mutation isolates, 13.3%(2/15) mutations appeared in inhA 5, 6.7%(1/15) in inhA 16, and 20%(3/15) in both katG and inhA. Conclusions The mutation of rpoB, katG and inhA genes in TB is highly correlated with its resistance to rifampin and isoniazid in Baise district, Guangxi autonomous region. The study will provide a basis for further understanding the anti-bacterium mechanism and quick diagnostic methods for drug-resistant tuberculosis.
10.A sequence investigation on Wallerian degeneration after pontine infarction with diffusion tensor imaging
Zhi-Jian LIANG ; Si-Run LIU ; Jin-Sheng ZENG ; An-Ding XU ; Xue-Ying LING ; Fang WANG ; Li LING ; Qing-Hua HOU ; Shi-Hui XING ;
Chinese Journal of Neurology 2005;0(11):-
Objective To investigate the Wallerian degeneration of neural fiber tract in medulla and bilateral middle cerebellar peduncle following pontine infarction and to explore its impacts on neurological recovery.Methods Fourteen patients with a recent unilateral pontine infarct underwent the diffusion tensor imaging(DTI)and evaluations with the NIH stroke scale(NIHSS),the Fugl-Meyer motor scale(FM), ataxia rating scale(ARS)and the Barthel index(BI)at the first week(W1),the fourth(W4)and twelfth week(W12)respectively.Mean diffusivity(MD)and fractional anisotropy(FA)were measured at pons, medulla and middle cerebellar peduncle.Fourteenth age and gender matched volunteers underwent a DTI were studied as controls.Results Compared with the matched regions in controls,the FA values of infarct side medulla and bilateral middle cerebellar peduncle in patients significantly decreased at W1,W4 and W12(on the infarct side of medulla:W1:0.43?0.01;W4:0.37?0.02;W12:0.30?0.02;on the infarct side of middle cerebellar peduncle:W1:0.50?0.01;W4:0.43?0.02;W12:0.35?0.04;on the opposite side of middle cerebellar peduncle infarction:W1:0.54?0.02;W4:0.52?0.03;W12:0.47?0.04,t values are 1.92 to 28.56,P0.05 respectively).The absolute value of percent reduction of FA in infarct side medulla and bilateral middle cerebellar peduncle was correlated negatively to the absolute value of percent change of NIHSS and BI score(P