Objective To explore the inhibitory effects of sulfated oligosaccharides PI-88 on the heparanase protein expression of human esophageal squamous cancer cell (ESCC) line TE-13, and to explore the effects of growth, angiogenesis and heparanase protein expression on ESCC xenografts of nude mice. Methods TE-13 cells were cultured and divided into three groups:group A (control group), group B (15 mg/L PI-88) and group C (30 mg/L PI-88). Heparanase protein expression of TE-13 cells was measured by Western blot assay after being cultured for 36 h. The ESCC suspension was injected subcutaneously in 10 BALB/c/nu mice to build up ESCC xenograft model. The model mice were divided randomly into observation group and control group (5 mice per group). The mice in observation group received 40 mg/(kg·d) PI-88. The mice in control group only received the same volume of saline at the same time. Both PI-88 and saline were daily administrated for 14 days. Every 2 days,the volume of xeongrafts were measured and the mice were executed at the 14th day. CD34 immunohistochemical staining was used to detect the micro vessel density (MVD) of xenografts. Western blot assay and immunohistochemical staining were used to detect the heparanase protein expression of xenografts. Results The heparanase protein expressions of TE-13 cells were significantly decreased in group B and group C than those of group A (P<0.001), with a kind of PI-88 dose-dependent manner. The volume, MVD and heparanase protein expression of xenografts were significantly lower in observation group than those of control group (P<0.05). Conclusion The heparanase protein expression in TE-13 cells can be inhibited by PI-88 in vitro and vivo. Furthermore, the growth and angiogenesis of ESCC xenografts were also inhibited by PI-88.

Objective To sum up the experience in the management of acute infrarenal abdominal aorta occlusion. Methods We retrospectively analyzed 34 cases of acute infrarenal abdominal aorta occlusion treated during a period of 18 years. Cell saver was used intraoperatively in five cases.Results Twelve cases died postoperatively in this group (35 3%), with acute post operative renal failure, metabolic acidosis and hyperkalemia being the main causes of death. No acute renal failure and metabolic acidosis occurred in all cases treated by cell saver during operation. Limbs were salvaged in 36 out of 44 extremities of patients surviving the surgical procedure. ConclusionsPrompt diagnosis, proper surgery and correct management of post operative complications are necessary to decrease mortality. Perioperative application of cell saver is recommended to eliminate metabolic wastes from the body.

Dopa-responsive dystonia (DRD) is a complex genetic disorder with either autosomal dominant or autosomal recessive inheritance, with autosomal dominant being more frequent. Autosomal dominant DRD is known to be caused by mutations in the GCH1 gene, with incomplete penetrance frequently reported, particularly in males. Here, we report a male patient with DRD caused by exon 1 deletion in the GCH1 gene inherited from the asymptomatic mother. The patient had an atypical presentation, notably with no dystonia, and underwent extensive workup for a myriad of neuromuscular disorders before a low-dose L-dopa trial and confirmatory genetic testing were performed. Our experience with this family highlights an atypical presentation of DRD and prompts us to consider the genetic complexity of DRD.

Objective To investigate the current status of hand hygiene(HH) among health care workers(HCWs) in clinical laboratories in medical institutions in Xi'an City.Methods HH status of HCWs in clinical laboratories in medical institutions in Xi'an was performed random on-the-spot sampling and monitoring.Results A total of 240 HH specimens of HCWs in clinical laboratories in 80 medical institutions in Xi'an City were collected, 127 detected results were qualified, the total qualified rate was 52.92%.The qualified rates of medical institutions were as follows: municipal hospitals 62.67%,workers' hospitals 55.95%,private hospitals 40.74%;comprehensive medical institutions 67.68%,specialized medical institutions 42.55%;tertiary medical institutions 79.63%(n=43),secondary and below medical institutions 45.16%(n=84),there were significant differences in HH qualified rate among HCWs in different types of medical institutions(all P<0.01).Of different HH detection items, detection rates of Escherichia coli and Staphylococcus aureus were 0.83% and 8.33% respectively.There were significant differences in HH compliance rates among HCWs of all age groups(χ2=9.103,P<0.05), HCWs aged≥50 years had the highest qualified rate of HH(71.43%), followed by those aged<30 years (67.82%),HCWs in 40~ year age group had the lowest HH qualified rate (39.66%).Conclusion The qualified rate of HH of HCWs in clinical laboratory of medical institutions in Xi'an City is low, it is necessary to enhance the procaution awareness of HCWs in clinical laboratories, strengthen quality control of HH, strictly implement standard hand-washing procedures to reduce occurrence of HAI.

To use the designed restriction enzyme assisted mutagenesis technique to perform rapid site-directed mutagenesis on double-stranded plasmid DNA. The target amino acid sequence was reversely translated into DNA sequences with degenerate codons, resulting in large amount of silently mutated sequences containing various restriction endonucleases (REs). Certain mutated sequence with an appropriate RE was selected as the target DNA sequence for designing mutation primers. The full-length plasmid DNA was amplified with high-fidelity Phusion DNA polymerase and the amplified product was 5' phosphorylated by T4 polynucleotide kinase and then self-ligated. After transformation into an E. coli host the transformants were rapidly screened by cutting with the designed RE. With this strategy we successfully performed the site-directed mutagenesis on an 8 kb plasmid pcDNA3.1-pIgR and recovered the wild-type amino acid sequence of human polymeric immunoglobulin receptor (pIgR). A novel site-directed mutagenesis strategy based on DREAM was developed which exploited RE as a rapid screening measure. The highly efficient, high-fidelity Phusion DNA polymerase was applied to ensure the efficient and faithful amplification of the full-length sequence of a plasmid of up to 8 kb. This rapid mutagenesis strategy avoids using any commercial site-directed mutagenesis kits, special host strains or isotopes.
Amino Acid Sequence ; Base Sequence ; DNA ; genetics ; DNA Restriction Enzymes ; genetics ; DNA-Directed DNA Polymerase ; genetics ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; methods ; Plasmids ; Receptors, Polymeric Immunoglobulin ; genetics

ObjectivesTo analyse the causes of occlusion of prosthetic grafts and to explore the appropriate modality of reoperation.MethodThe clinical data of 47 patients with occlusive prosthetic grafts were analyzed retrospectively. The procedure of treatment for patients with different causes of occlusive graft included thrombectomy alone in 9 cases, thrombectomy and anastomotic plasty in 16 cases, new bypass using prosthetic graft and saphenous autograft in 22 cases. ResultsThe reoccurrence of graft occlusion in patients undergoing thrombectomy alone or thrombectomy and anastomotic plasty was 67% and 56% respectively, which was siginificantly higher than that of 9% in patients treated by a new arterial bypass with prosthetic grafts and saphenous autografts. ConclusionsThe result of new bypass for occlusive prosthetic graft is superior to thrombectomy alone or thrombectomy and anatomotic plasty.

Abstract: To express human-mouse chimeric IgA antibody directed against H5N1 virus, an anti-H5N1 chimeric IgA antibody gene was constructed by joining the light and heavy chain variable region genes and the corresponding signal peptide coding sequences of the anti-H5N1 mouse monoclonal antibody H5N1-HA with the coding sequences of the constant region of the human IgA2 heavy chain and Kappa chain respectively. Then the full-length chimeric light and heavy chain expressing plasmids pEF-IGHA9 and pEF-IGK9 were constructed and transfected into the CHO/dhfr cells. The chimeric IgA antibody expression was confirmed by ELISA, SDS-PAGE and Western blotting. The successful expression of this anti-H5N1 chimeric IgA may help to provide a stand for developing passive immunological agents for H5N1 virus infection prophylaxis.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibodies, Viral ; genetics ; CHO Cells ; Chimerism ; Cricetinae ; Cricetulus ; Humans ; Immunoglobulin A ; genetics ; immunology ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Mice ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology