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.

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.

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.

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.

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.

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.

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.

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.

To establish a platform for high throughput screening and in vitro evaluating novel metabolic enzyme-targeted inhibitors towards anti-malarial drugs, a lactate dehydrogenase gene of Plasmodium falciparum (PfLDH) was amplified from the Hainan isolate FCC1/HN. The fusion expression vectors, pGEX-2TK and pET-29a( + ), were utilized to introduce the PfLDH gene into strains of Escherichia coli, BL21 and BL21 (DE3), for over-expression. Consequently, the enzymatic activity of PfLDH was successfully detected in the suspension of lytic bacteria. The PfLDH gene cloned in pGEX-2TK was mainly expressed as inclusion bodies, while the same gene cloned in pET-29a( + ) was nearly expressed in a soluble form of PfLDH, demonstrating the latter vehicle might be more suitable for the large-scale preparation of recombinant PfLDH. Furthermore, according to the electrophoregram of SDS-PAGE and the sequencing data, a series of truncated PfLDH sequences generated randomly from gene amplification were screened and cloned, from which four pre-matured genes with a terminator mutation, PfLDH-delta271, -delta236, -delta167 and -delta53 coding for 45, 80, 149 and 263 amino acid residues, were individually recovered. Through the gene expression and enzymatic activity measurement, the effect of pre-matured terminator mutation on the activity of PfLDH was evaluated, which should pave the way for probing the relationship between structure and function of PfLDH.
Animals ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Inclusion Bodies ; genetics ; metabolism ; L-Lactate Dehydrogenase ; genetics ; metabolism ; Plasmodium falciparum ; enzymology ; genetics ; Protozoan Proteins ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Solubility

Objective :To study and observe application value of 256 slice spiral CT coronary imaging (MSCT) for di‐agnosing coronary artery stenosis in aged patients .Methods :A total of 95 aged patients suspecting coronary artery stenosis treated in our hospital were selected .They received MSCT and coronary angiography (CAG) respectively . Inspecting outcomes of above two diagnostic methods were comprehensively compared .Results :(1) CAG identified 91 positive cases and four negative cases in diagnosing coronary artery stenosis ,while it＇s 89 positive cases and six negative cases for MSCT .The diagnostic outcomes of two methods were highly consistent (Kappa＝ 0.789 , P＝0.001 ) ,suggesting MSCT can achieve the diagnostic effect of CAG ; (2) Sensitivity ,specificity ,positive predictive value ,negative predictive value and accuracy of MSCT diagnosing coronary artery stenosis was 94. 51%,25.00%, 96. 63%,16.67% and 91.58% respectively ,suggesting diagnostic outcomes of MSCT possessed high accuracy ; (3) There was no significant difference in judgment of coronary artery stenosis degree between MSCT and CAG , P＝0.524. Conclusion :The diagnostic accuracy of 256 slice MSCT is high in aged patients with coronary artery stenosis , which is almost consistent with that of the gold standard‐CAG .The conduction is simple and it＇s noninvasive ,which can be extended in clinic ;but it′s specificity is compare less ,must pay suitable intension