[Application of anticoagulating medicine after surgery of cavity of pelvis and abdomen].
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Abdomen↗
Biomedical subjects
Qing-hua Wu
Publications and source records attributed to Qing-hua Wu.
Long-term results of elective open repair for abdominal aortic aneurysm.
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Animals↗
[Preparation of small interfering RNA expression cassette based on PCR technique].
OBJECTIVE: To modify the current PCR-based method for rapid and efficient preparation of small interfering RNA (siRNA) expression cassette to improve the efficiency of RNA interference. METHODS: The U6 promoter sequence was amplified by PCR using the genomic DNA of K562 cells as the template, and cloned into pMD18-T vector which served as the template for further PCR amplification with the primers on the plasmid. The amplified product was directly used as the template for preparing siRNA expression cassette. The siRNA expression cassette targeting p53 gene was amplified, verified by sequencing, and transfected into SH-SY5Y cells. After a 48-hour transfection, the cells were harvested and the total RNA was for RT-PCR for evaluating the effect of RNA interference. RESULTS: The sequencing result confirmed the correct U6 promoter sequence cloned from K562 cells. After transfection of SH-SY5Y cells for 48 h with siRNA expression cassette, the p53 gene expression was inhibited at the mRNA level in comparison with the control cells as demonstrated by RT-PCR detection. CONCLUSION: The siRNA expression cassette prepared using the established method described hereby can be well applicable in RNA interference research.
Gene Silencing↗
[Experiences of retroperitoneal approach for aortoiliac reconstruction].
OBJECTIVE: To summarize experiences of aortoiliac reconstruction through retroperitoneal approach. METHODS: Twenty-eight patients underwent retroperitoneal aortoiliac reconstructions, including aortic aneurysmectomy with graft replacement, aortic endarterectomy with patch angioplasty, thoraco-abdominal aortic bypass, resection of retroperitoneal mass with ilio-femoral bypass, iliac aneurysmectomy with aorto-external iliac artery bypass, removal of aortoiliac foreign body, common iliac endarterectomy, aorto (ilio)-femeral bypass and common ilio-femo-popliteal bypass. Drainage tubes were placed retroperitoneally in 24 cases. RESULTS: All operations in this group were successful without perioperative death. The volume of intra-operative bleeding was 100-400 ml (mean 240 ml). Blood transfusion were employed in 2 cases. Retroperitoneal drainage was 50-170 ml (mean 85 ml). Naso-gastric tubes were removed 28 h on average after operation. All patients recovered uneventfully except that cardiac insufficiency, stress ulcer and retroperitoneal hematoma were present in 3 patients respectively. Twenty-two patients were followed up from 3 months to 2.5 years. One patient died of AMI 2 years after operation. One patient receiving ilio-femo-popliteal bypass was found to have occlusion of femo-popliteal segment of prosthetic graft. One patient developed brain hemorrhage 1.5 years postoperatively. All the other followed-up patients were living well. CONCLUSION: Retroperitoneal approach, not violating the peritoneal cavity, offers certain physiological advantages associated with minimal disturbance of gastrointestinal and respiratory function, thereby decreasing respiratory complications and postoperative ileus, avoiding intra-abdominal adhesions with their attendant risk of early and late small bowel obstruction. It proved to be a simple and safe alternative for surgical treatment of aortoiliac diseases.
Adult↗
[Comparison of two amine-modified chemical platforms for DNA microarray preparation].
OBJECTIVE: To study two amine modification procedures for DNA microarray preparation based on polymeric coatings. METHODS: One of the proposed approaches utilized poly-amine coating of silanized slides activated by 1,4-phenylene diisothiocyanate, and the other employed acrylic acid-co-acrylamide copolymer and 1-(3-dimethylamino propyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide as the coating agents and activator, respectively. The modified slides were used for preparing lambda phage DNA microarrays, whose properties were analyzed by hybridization. RESULTS: Formation of dendrimeric structure and polymer was observed on the surface of the slides. The signal spots in uniform, steady and regular shape, in comparison with the commercial CMT-GAPS slides, indicated successful manufacture of the microarrays. CONCLUSION: The two platforms are suitable for microarray preparation, and the method of acrylic acid-co-acrylamide copolymer modification is more preferred.
Acrylamides↗
[Amplification and cloning of the N gene of SARS-associated coronavirus].
OBJECTIVE: To amplify and clone the N gene of severe acute respiratory syndrome-associated coronavirus. METHOD: Using primer Premier 5.0 software, two pairs of nested PCR primers were designed to amplify the N gene. After purification, the amplified products were cloned into pMD18-T vectors, and the positive clones with the inserted fragments were identified by sequence analysis. RESULTS: The amplified products was about 1 375 bp in length, and sequence analysis demonstrated that the N gene fragments had been successfully inserted into pMD18-T vectors. CONCLUSION: The successful amplification and cloning of N gene facilitates further investigation of the expression of the N protein and study of its structure and functions.
Base Sequence↗
Design and preparation of oligonucleotide microarray for vaccinia virus detection.
OBJECTIVE: To study the preparation of oligonucleotide microarray for detecting vaccinia virus. METHODS: Oligonucleotide probes were designed and synthesized according to the specific genes of vaccinia virus. Sample DNA of the virus and the negative control sample were obtained and labeled by restriction display technique, followed by hybridization to the oligonucleotide microarray and scanned by Agilent scanner. RESULTS: Strong hybridization signals were detected from the viral DNA hybridized with the microarray, but were absent in the negative sample when positive probes were not used. CONCLUSION: Distinct differences in the hybridization signals between the virus sample and negative sample and between the samples obtained in different phase of infection demonstrate high specificity and sensitivity of the microarray for vaccinia virus detection.
Base Sequence↗
[Isolation of the target gene from cDNA restriction fragments using 70-mer oligo microarray].
OBJECTIVE: To study the method for using a 70-mer oligo microarray as the probe to isolate target genes from the cDNA restriction fragments. METHOD: Samples of Saccharomyces cerevisiae mRNA was extracted after heat shock culture and reversely transcribed into the double-stranded cDNAs, which were prepared into restriction cDNA fragments using restriction display (RD) method. The microarray was printed using a single 70-mer specific oligo designed to according to the SSA1 gene of yeast. The cDNA restriction fragments were labeled by PCR method with the Cy5 universal primer before hybridization with the microarray. The microarray was stripped after washing and scanning, and the strip solution was collected for another round of PCR amplification using the universal primer without fluorescence. The PCR product was then cloned into PUC18 T vector and transformed into to E.coli JM109 cells for amplification, and the plasmids were extracted and sequenced for identification. RESULTS: BLAST results showed that the target gene was cloned successfully. CONCLUSION: The target gene can be isolated directly using the 70-mer oligo microarray as the probe from the cDNA fragments prepared by RD method, without the necessity of building a cDNA library. This method can also be used in further research to acquire the differentially expressed genes after the oligo microarray hybridization.
Base Sequence↗
[Oligonucleotide microarray preparation using enhanced poly-L-lysine glass slides].
OBJECTIVE: To modify conventional poly-L-lysine coating for oligonucleotide microarray preparation so as to enhance the sensitivity of the microarray. METHOD: The proposed chemical approaches included silanizing the slides with 3-glycid-oxypropyltrimethoxysilane (GOPS) after cleaning, followed by slide coating with polymers (poly-L-lysine) that was covalently bound to the modified glass. Subsequent attachment of the oligonucleotide to the modified slide surface was achieved after 1,4-phenylene diisothiocyanate (PDITC) activation of the surface. Various experiments were carried out, such as the immobilization efficiency and hybridization assays to test the modified slides, which were then used tentatively in the preparation of microarrays for SARS coronavirus detection. RESULTS: The improved surface had high immobilization efficiency, good uniformity and satisfactory hybridization efficiency, better than those slides with conventional poly-L-lysine coating. In addition, such modified slides rendered the microarrays more resistant to consecutive probing/stripping cycles. CONCLUSION: The modified slide surface is satisfactory to immobilize unmodified oligonucleotide by covalent binding, which enhances not only the sensitivity of the prepared oligonucleotide microarray but also the binding of the oligonucleotide to the slide surface.
DNA, Viral↗
[Operative outcome and experience in arteriovenous shunt at different sites].
OBJECTIVE: To summarize and analyze the influencing factors of both success and half-year patency rate of radial artery-cephalic vein arteriovenous fistula. METHODS: A retrospective analysis was made on the clinical data of 102 patients with chronic renal failure undergoing radial artery-cephalic vein arteriovenous shunt for hemodialysis during 7 years and the influence of different factors: sex, age, primary disease, incision site, and situations of vessels on the operative success rate and the half-year patency rate of the fistula was compared. RESULTS: No significant difference in the initial success rate of operation was found between the operations performed at the two different incision sites: at the wrist and at anatomist's snuffbox. However, the operation at the wrist was better than that at anatomist's snuffbox with regard to the half-year patency rate. Both operative success rate and half-year patency rate were not influenced by patient's sex, age, and primary disease. Anastomotic hyperplasia, condition of blood vessel and anastomotic technique were important factors influencing the operative failure and fistula occlusion. CONCLUSION: Correct selection of incision site is of key importance for both operative success and maintenance of half-year patency of radial artery-cephalic vein arteriovenous fistula. In addition, anastomotic hyperplasia, diameter and intimal smoothness of selected blood vessel, condition of run-off vessel, operative technique, anastomotic angle, distance between artery and vein, and patient's self-protection are also considered important influencing factors.
Adult↗
[Application of percutaneous transluminal angioplasty and stent in the treatment of subclavian steal syndrome].
OBJECTIVE: To explore the clinical effect of percutaneous transluminal angioplasty (PTA) and stent on the subclavian steal syndrome. METHODS: The clinical results of PTA and stent in the treatment of subclavian steal syndrome in 78 patients were analysed. RESULTS: 100% interventional procedure success rate was achieved with a mean systolic arterial pressure difference of <or= 10 mm Hg between both upper limbs and normalization of radial pulses. No major complications occurred. During a period of 27-month followup 66 patients were free of adverse events except fail of followup in 12 patients. CONCLUSION: PTA with stent placement is a good method in the treatment of subclavian steal syndrome. It is a safe, easy to be performed, minimally invasive procedure with good clinical results.
Adolescent↗
[Analysis with DNA chips of the changes of gene expressions in K562 cells in response to As2O3 treatment].
OBJECTIVE: To investigate differential gene expression in apoptotic cells induced by As(2)O(3), and identify novel apoptosis-related genes. METHOD: Apoptosis of K562 cells cultured in RPMI 1640 medium supplemented with 10 % calf serum was induced by As(2)O(3). Total RNA of the apoptotic and normal cells were then extracted, purified and subject to reverse transcription into first-strand cDNA, labeled with Cy3/Cy5. Placenta DNA microarrays containing 348 DNA fragments were used to analyze the changes in gene expressions in the cells treated with As(2)O(3). RESULT: Eleven differentially expressed genes were identified in the apoptotic cells in comparison with the normal cells, 3 of which were associated with apoptosis, while the others were related to cell growth and proliferation. CONCLUSION: The placenta DNA microarrays we constructed may well apply to the analysis of the differentially expressed genes.
Antineoplastic Agents↗