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To explore the expression and clinical significance of thymidine phosphorylase/platelet-derived endothelial cell growth factor (PD-ECGF) in renal cell carcinoma.The expression of PD-ECGF in 76 specimens of renal cell carcinoma from June 2002 to December 2006 was examined by EnVision immunohistochemistry. The correlation of the expression of PD-ECGF with the clinicopathological characteristics and prognosis in renal cell carcinoma patients were analyzed.Among 76 renal cell carcinoma patients, 41 patients had a high-level expression of PD-ECGF while another 35 patients a low-level expression. The expression of PD-ECGF was positively correlated with tumor stage and histological grade. The 5-year overall survival rate was significant lower in the patients with a high-level expression of PD-ECGF (60.98% vs 94.29%, P = 0.000). Univariate analysis showed that the characteristics with the prognostic significance for survival were tumor stage, lymph node metastasis, distant metastasis, histological grade and the expression of PD-ECGF. Multivariate analysis showed that tumor stage, histological grade and the expression of PD-ECGF were independent prognostic factors (OR: 17.971, 9.702, 6.790, 95%CI: 3.010 - 107.290, 1.835 - 51.305, 1.156 - 39.894).The expression of PD-ECGF may play an important role in the development and invasion of renal cell carcinoma. An elevated level of PD-ECGF is probably correlated with the prognosis of renal cell carcinoma.

In recent years, researchers have identified a large number of complex diseases/traits-associated genetic variants by performing genome-wide association studies (GWAS), which may provide important clues on understanding the mechanisms of related diseases. However, GWAS has its own limitations in terms of being false positive, false negative results, very few SNPs located in the functional areas and insensitive to detect rare and structural variations, which results in the application limitation of this method. With the development of the next-generation sequencing technology, whole genome and exome sequencing developed rapidly and provide an opportunity for us to deal with the problem caused by GWAS. This high-throughput sequencing technology is applied for sequencing the exome (1% of genome) to discover most of the diseases-related variations in exons. Furthermore, it is highly effective to detect common and rare variations. Due to these advantages, exome sequencing has become a powerful and efficient strategy for identifying the genes responsible for mendelian disorders and complex diseases, which will be very helpful for the diseases clinical diagnosis.

Background/Aims: Preventing internal hemorrhage extends the lifespan of rats with chronic bile duct ligation (CBDL), a common animal model of portal hypertension. We investigated hemodynamics during the early and late stages of cirrhosis caused by CBDL. We also evaluated the hemodynamic influence of NO, which is the chief vasodilator in hyperdynamic syndrome, by administration of an NO synthase inhibitor (NG-nitro-L-arginine methyl ester: L-NAME; 10 mg/kg). Animals/Methods: In 24 Sprague-Dawley rats (9 sham rats and 15 CBDL rats), hemodynamics were assessed under conscious and unrestrained conditions 4 and 8 weeks after surgery. Before and 30 minutes after L-NAME administration, the cardiac index (CI) and regional blood flow were measured with the reference sample method using 141Ce- and 113Sn-microspheres (15 μm in diameter). Results: A hyperdynamic systemic circulation and splanchnic hyperemia were observed after CBDL, and these changes increased with the progression of cirrhosis. L-NAME significantly diminished the hyperdynamic circulation and also reduced splanchnic hyperemia. In 4-week CBDL rats, a low hemoglobin concentration made an important contribution to the hyperdynamic circulation, and the portal collateral system collapsed when inflow to the portal territory was reduced by L-NAME treatment. In 8-week CBDL rats, systemic hemodynamics were closely linked to both the splanchnic circulation and the renal circulation before and after L-NAME administration, apart from hepatic artery blood flow. Conclusion: The distinctive hemodynamic changes of portal hypertension were found in 8-week CBDL rats. Thus, 8-week CBDL rats may be a better animal model of human portal hypertension than 4-week CBDL rats.

In past five years, genome-wide association study (GWAS) has proven to be a powerful and efficient study design in identifying genetic variants associated with complex diseases and traits. Many studies have been performed to explore the genetic basis of various complex diseases and traits. Genetic breakthroughs have been achieved in complex diseases and traits, such as oncology, diabetes, cardiology, psychiatric disorders, autoimmune and immune related diseases, height, weight, blood lipid, and pigmentation. Up to September 11th 2010, nearly 200 complex diseases and traits have been investigated by this approach, and more than 3000 diseases and/or traits related variants have been identified. In this review, we present an overview of GWAS in complex diseases and traits.

Host immune responses are critical steps for carcinogenesis. Single nucleotide polymorphisms (SNPs) in immunoregulatory genes may influence gastric cancer risk. We performed a genotyping analysis for immunoregulatory genes in 311 gastric cancer cases and 425 controls from a Chinese population. We found that there were significant differences of E-selectin variant rs5361 (AC) and FCGR2A variant rs1801274 (TC) between cases and controls (P = 0.022 and P = 0.0001, respectively). Logistic regression analysis indicated that genotype of E-selectin rs5361AC increased the risk of gastric cancer significantly (P = 0.026, adjusted Odds ratio (OR) = 2.84, 95% confidence interval (CI) = 1.13-7.12). C allele of E-selectin rs5361 showed a significant increased frequency in cases (P = 0.023). However, the E-selectin variant did not affect the protein expression. E-selectin protein was observed not only in tumor interstitial vascular endothelial cells, but also in gastric cancer cells at primary and metastatic sites. The protein was associated with clinicopathological characteristics of gastric cancer, such as age (P = 0.008), tumor size (P = 0.027), differentiation (P = 0.000), and tumor-node-metastasis (TNM) stage (P = 0.006). CT and CC + CT genotypes of FCGR2A variant rs1801274 increased gastric cancer risk (P = 0.000, adjusted OR = 1.92, 95%CI = 1.36-2.72; P = 0.003, adjusted OR = 1.68, 95%CI = 1.20-2.35, respectively). Interleukin-4 receptor (IL-4R) variant rs2107356 presented negative correlations to E-selectin variant rs5361 and FCGR2A variant rs1801274 (P = 0.035 and P = 0.023) in conferring susceptibility to gastric cancer. We concluded E-selectin variant rs5361 and FCGR2A variant rs1801274 were significantly associated with gastric cancer risk. Expression of E-selectin protein would promote progression of gastric cancer.

Leber's hereditary optic neuropathy (LHON) is a maternally transmitted disease. Clinically, no efficient assay protocols have been available. In this study, we aimed to develop an oligonucleotide biochip specialized for detection of known base substitution mutations in mitochondrial DNA causing LHON and to investigate frequencies of LHON relevant variants in Anhui region of China. Thirty-two pairs of oligonucleotide probes matched with the mutations potentially linked to LHON were covalently immobilized. Cy5-lablled targets were amplified from blood DNA samples by a multiplex PCR method. Two kinds of primary mutations 11778 G > A and 14484 T > C from six confirmed LHON patients were interrogated to validate this biochip format. Further, fourteen Chinese LHON pedigrees and twenty-five unrelated healthy individuals were investigated by the LHON biochip, direct sequencing and pyrosequencing, respectively. The biochip was found to be able efficiently to discriminate homoplasmic and heteroplasmic mtDNA mutations in LHON. Biochip analysis revealed that twelve of eighteen LHON symptomatic cases from the 14 Chinese pedigree harbored the mutations either 11778G > A, 14484T > C or 3460G > A, respectively, accounting for 66.7%. The mutation 11778G > A in these patients was homoplasmic and prevalent (55.5%, 10 of 18 cases). The mutations 3460G > A and 3394T > C were found to co-exist in one LHON case. The mutation 13708G > A appeared in one LHON pedigree. Smaller amount of sampling and reaction volume, easier target preparation, fast and high-throughput were the main advantages of the biochip over direct DNA sequencing and pyrosequencing. Our findings suggested that primary mutations of 11778G > A, 14484T > C or 3460G > A are main variants of mtDNA gene leading to LHON in China. The biochip would easily be implemented in clinical diagnosis.

The propensity of an amino acid to form an alpha helix in a protein was determined by multiple amino substitutions at positions 44 and 131 in T4 lysozyme. These positions are solvent-exposed sites within the alpha helices that comprise, respectively, residues 39 to 50 and 126 to 134. Except for two acidic substitutions that may be involved in salt bridges, the changes in stability at the two sites agree well. The stability values also agree with those observed for corresponding amino acid substitutions in some model peptides. Thus, helix propensity values derived from model peptides can be applicable to proteins. Among the 20 naturally occurring amino acids, proline, glycine, and alanine each have a structurally unique feature that helps to explain their low or high helix propensities. For the remaining 17 amino acids, it appears that the side chain hydrophobic surface buried against the side of the helix contributes substantially to alpha helix propensity.

To investigate the diagnosis, treatment and outcomes of bone lymphangioma associated with spinal deformity in children.Eight cases of children with bone lymphangioma associated with spinal deformity at our hospital from 1983 to 2010 were reviewed and analyzed with the following criteria: clinical manifestations, imaging features, histopathological characteristics and therapeutic options. The outcomes of early intervention treatment were assessed. The key diagnostic points and therapeutic principles were summarized on the basis of our own as well as international and domestic experiences.Paraplegia developed in 2 cases because of delayed treatment. The remaining 6 cases were followed up for 10 months to 2 years with satisfactory effects. By curettage lesions, bone grafts, intralesional injection of bleomycin and pedicle screw-stick system maintaining the spinal stability, bone lymphangioma became smaller or disappeared and spinal deformity was effectively controlled. The mean scoliosis and kyphosis correction rates were 57.0% and 58.4% respectively.Bone lymphangioma in children is rare. If costa and vertebrae are involved, spinal deformity will progressively develop and lead eventually to paraplegia. With an early diagnosis and the administration of proper drugs, bone lymphangioma can be effectively controlled. Surgery for the stability of spinal deformity will effectively prevent the occurrence of paraplegia.

To compare the postoperative depths of the coagulation zones and pathological changes between bipolar transurethral resection of the prostate with plasmakinetic energy (PKRP) and monopolar transurethral prostatectomy (TURP) in canines.Twenty-five male dogs were randomly divided into a PKRP group (n = 12), a TURP group (n = 12) and a sham-operation control group (n = 1). The dogs were sacrificed, their prostates harvested at 0 week (immediately after surgery), 1 week, 2 weeks and 8 weeks postoperatively and sectioned for pathologic analysis and measurement of the coagulation zones.At 0, 1 and 2 weeks after the operation, the coagulation depths were (237.73 +/- 20.12) microm, (113.03 +/- 16.65) microm and (106.01 +/- 16.36) microm in the PKRP group, and (200.75 +/-19.34) microm, (129.46 +/- 17.81) microm and (116.04 +/- 25.67) microm in the TURP group (P0.01). At 8 weeks, the coagulation zones completely peeled off and the wounds were covered by regenerated urothelial in both of the groups. At 0, 1, 2 and 8 weeks, different inflammatory reactions were observed in the prostates of the PKRP and TURP groups, with some glandular lumens beneath the coagulation zones expanded and epithelia damaged. However, none of these phenomena occurred in the sham-operation control group.Pathologically, PKRP and TURP inflicted basically similar effects on the prostate of the canine. However, the coagulation zone was deeper intraoperatively and became thinner postoperatively with the former than with the latter, which suggests that PKRP causes less bleeding and less penetrative thermal damage than TURP.

The multifunctional protein p100 is a vital transcriptional regulator that increases gene transcription by forming a physical bridge between promoter-specific transcription factors and the basal transcription machinery. To identify potential signal transduction pathways in which human p100 acts as a coregulator and to find target promoter regions that may interact with p100, we performed a promoter microarray assay called chromatin immunoprecipitation-guided ligation and selection (ChIP-GLAS). From this assay, we determined that a set of promoter fragments, including several factors in the transforming growth factor beta (TGF-β) signaling pathway, exhibited interaction with p100. The ChIP-GLAS data were validated by RT-PCR assessing the mRNA expression of various factors in the TGF-β signaling pathway in cell lines.

BACKGROUND To examine the relation between demographic factors and symptom type in the use of gynecological health services in rural China. METHODS Married women aged 19 to 45 years from three rural communities in Anhui province, central China, were invited to participate in a structured interview in the summer of 2006. They provided information on gynecological symptoms, healthcare-seeking behaviour and socio-demographic characteristics. Risk factors were analyzed using logistic regression. RESULTS 860/1221(70.4%) reported at least one gynecological symptom during the previous year, with 485 (39.7%) reporting three or more. Of the women with symptoms, 36.7% sought treatment during the previous year. Younger women and those with multiple symptoms were more likely than others to seek treatment. Women with abnormal vaginal bleeding or discharge were more likely to delay seeking treatment. Years of education were strongly related to seeking treatment. More highly educated women and women with a higher household income were more likely than others to seek treatment at the highest level (county or city hospital) of the tertiary healthcare system rather than at a village clinic or township hospital. Women who did not seek treatment were more likely to report that they saw no need than to say that they could not afford care. CONCLUSION There may be a misperception of the need for, and utility of, treatment for gynecological symptoms, particularly in more disadvantaged women. Interventions should both address women's negative perceptions and reinforce the capacity of the local health facilities to ensure effective care.

Interleukin 13 (IL-13) is a key cytokine involved in the regulation of inflammatory, immune responses, and cell differentiation. The present study was to investigate the effect of IL-13 on the expression of glycoprotein IIb (GPIIb), a megakaryocytic gene, in Dami cells (human megakaryoblastic leukemia cell line) and HEL cells (human erythroleukemic cell line, which has both erythroid and megakaryocytic markers). Furthermore, it addresses the mechanisms governing the regulation of GPIIb expression by IL-13. The molecular responses of Dami cells and HEL cells to IL-13 treatment were analyzed by RT-PCR, Western blot, chromatin immunoprecipitation (ChIP) and flow cytometry analysis. We show that IL-13Rα1 and IL-4Rα are expressed in Dami cells and HEL cells. The expression of GPIIb was significantly upregulated at the mRNA and protein levels by treatment with IL-13. Moreover, IL-13 induced phosphorylation of signal transducer and activator of transcription 6(STAT6). By using a STAT6-specific antibody and PCR primers designed to yield a product, which encompasses the STAT6 binding site of the GPIIb promoter, we have shown the binding of the IL-13-mediated activation of STAT6 to the promoter of GPIIb gene. These results broaden the involvement of IL-13 into megakaryocyte differentiation by STAT6 pathway.

To investigate the relationship between condyle movement and temporomandibular disorders (TMD) in patients with Class II division 1 malocclusion.Twenty patients (from 11 to 12 years old) with Class II division 1 malocclusion before treatment were collected. Computer aided diagnosis axiograph (CADIAX) and magnetic resonance images (MRI) were used to analyze the condyle movement between disc displacement and normal groups.The sensitive values were found in open/close process in patients with disc displacement: Y [Left: (0.32 +/- 0.10) mm, Right: (-0.91 +/- 0.49) mm ], Z [Left: (4.20 +/- 0.70) mm, Right: (3.44 +/- 0.21) mm], sagittal condylar inclination (SCI) [Left: (32.48 +/- 7.70) degrees , Right: (33.47 +/- 12.60) degrees ] and horizontal condylar inclination (TCI) [Left: (-2.60 +/- 2.02) degrees , R: (-9.23 +/- 5.58) degrees ], and those items showed significant difference between two groups.The side shift of condyle movement in maximum open/close process might be the inducement of disc displacement. It was revealed that the changes in condyle movement could give useful information in early stage of functional treatment.

The present study was purposed to investigate the inhibition effect of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) on growth of RPMI8226 cells and adhesion between RPMI8226 cells and bone marrow stroma cells (BMSC), and to explore its mechanism as well. The inhibition effects of TRAIL on cells growth and adhesion were assayed by MTT; cell apoptosis was detected by Annexin V and PI; expression of genes bax, bcl-2, mcl-1, CARP1, CARP2, XIAP and cFLIP were determined by semi-quantitative RT-PCR; apoptosis-related protein expression was detected by Western blot. The results showed that TRAIL inhibited the proliferation of RPMI8226 cells in dose- and time-dependent manners. TRAIL induced apoptosis in RPMI8226 cells, the expression level of genes bcl-2, mcl-1, CARP1, CARP2, XIAP and cFLIP decreased, while the expression level of Bax increased, but the expression level of caspase-3 and NF-kappaB P65(RelA) proteins decreased. Moreover, TRAIL up-regulated the expression level of adherent molecule CXCR4 in RPMI8226 cells significantly. It is concluded that TRAIL up-regulated the expression level of adherent molecule CXCR4 in RPMI8226 cells significantly, and induced the apoptosis of RPMI8226 cells. Growth inhibition effect of TRAIL on RPMI8226 cells is in dose- and time-dependent manners.

We performed a genome-wide association study of esophageal squamous cell carcinoma (ESCC) by genotyping 1,077 individuals with ESCC and 1,733 control subjects of Chinese Han descent. We selected 18 promising SNPs for replication in an additional 7,673 cases of ESCC and 11,013 control subjects of Chinese Han descent and 303 cases of ESCC and 537 control subjects of Chinese Uygur-Kazakh descent. We identified two previously unknown susceptibility loci for ESCC: PLCE1 at 10q23 (P(Han combined for ESCC) = 7.46 x 10(-56), odds ratio (OR) = 1.43; P(Uygur-Kazakh for ESCC) = 5.70 x 10(-4), OR = 1.53) and C20orf54 at 20p13 (P(Han combined for ESCC) = 1.21 x 10(-11), OR = 0.86; P(Uygur-Kazakh for ESCC) = 7.88 x 10(-3), OR = 0.66). We also confirmed association in 2,766 cases of gastric cardia adenocarcinoma cases and the same 11,013 control subjects (PLCE1, P(Han for GCA) = 1.74 x 10(-39), OR = 1.55 and C20orf54, P(Han for GCA) = 3.02 x 10(-3), OR = 0.91). PLCE1 and C20orf54 have important biological implications for both ESCC and GCA. PLCE1 might regulate cell growth, differentiation, apoptosis and angiogenesis. C20orf54 is responsible for transporting riboflavin, and deficiency of riboflavin has been documented as a risk factor for ESCC and GCA.

The idiotypic determinant of surface immunoglobulin of B-cell lymphoma, as a tumor-specific antigen, has proved to be able to induce immune responses. To analyze whether an idiotypic vaccine fused with cytokine can elicit more effectively protective antitumor immunity, an eukaryotic expression plasmid was constructed, which encoded the fusion gene of single-chain variable fragment as a tumor specific antigen against B-cell lymphoma with monocyte chemotactic protein-3 (MCP3) as immunogen to elicit T-cell-dependent protective antitumor immunity, and EGFP (Enhanced Green Fluorescent Protein) gene as a marker to trace the survival, growth, differentiation and expression of the former exogenetic genes. The cDNAs for immunoglobulin (Ig) VH and IgVL were amplified by RT-PCR and assembled into the single-chain variable fragment (scFv) connected with a (Gly(4)Ser)(3) linker by recombinant PCR method. Then, the fragments of scFv and MCP3 were ligated with a NDAQAPKS spacer by the same method. The results showed that the fusion genes of scFv and MCP3-scFv were inserted into an eukaryotic expression vector pTARGET, and EGFP was cloned into the downstream of scFv and MCP3-scFv respectively. Finally the constructed plasmids were confirmed by sequencing and restriction analysis. In conclusion, a tumor-derived idiotypic DNA vaccine, encoding the fusion gene of single-chain variable fragment and monocyte chemotactic protein-3 (MCP3) to elicit a T-cell dependent, antitumor immunity, and the EGFP gene was inserted correctly. The DNA vaccine could be used for further study of DNA vaccine against B cell lymphoma in vivo.

Psoriasis is a common autoimmune and hyper proliferative skin disease, characterized by thick, silvery scale patches. Numerous family studies have provided compelling evidence of a genetic predisposition to psoriasis, although the inheritance pattern is unclear. However, few of these studies have achieved consistent results, except for the MHC locus, a problem frequently encountered in the investigation of complex disease. Using high-throughput techniques to genotype hundreds of thousands of single nucleotide polymorphisms explore their relationship with phenotypes, genome-wide association studies (GWAS) are now proven to be a powerful approach for screening the susceptibility genes (loci) of complex disease. Recently, three GWAS on psoriasis published in Nature Genetics have provided us with many novel clues concerning disease pathogenesis, in both immune and non-immune pathways. The MHC locus (HLA-Cw6 and other MHC variance), the major locus involved in the immune reactions of human immune disease, has consistently been shown to be associated with psoriasis, both in previous linkage and present GWAS. IL-12B and IL23R, which are the two non-MHC genes with highly associated evidence with psoriasis in multiple studies performed so far and potent cytokines with complex biological activities, should be of great importance in the pathogenesis of psoriasis. Recent clinical trials, in which anti-IL-12p40 antibodies were used for the treatment of psoriasis, have provided further evidence of the role of IL-12/23 in the pathophysiology of psoriasis,and highlighted a new road of treatment for psoriasis. In 2008,we performed the first large GWAS in the Chinese population and identified a novel susceptibility locus within the late cornified envelope (LCE) gene cluster: LCE3A and LCE3D on chromosome 1q21, with conclusive evidence (rs4085613, p(combined)=6.69*10(-30); odds ratio=0.76). Meanwhile, another group also identified a deletion comprising and LCE gene cluster of LCE3B and LCE3C, which is significantly associated with a risk of psoriasis in Spain, Netherland, Italy and USA. Both of these independent studies provided substantial association evidence for the LCE genes involved in the pathogenesis of psoriasis. The LCE genes encode the stratum-corneum proteins of the cornified envelope, which plays an important role in epidermal terminal differentiation. As we know, psoriasis is a disease of interfollicular epidermis and rapid keratinocyte proliferation may cause the production of parakeratotic keratinocytes in psoriatic skin and, thus, the formation of poorly adherent stratum corneum, which in turn results in the characteristic scale or flakes of psoriasis lesions. Although some of the highlighted genes are already targeted by effective psoriasis therapies, others could become future targets for treatments,especially for the LCE genes, which will be very useful for unlocking new drug targets and tailored treatments for this painful, disfiguring skin disease. Meanwhile larger samples and improved strategy for identification of other susceptibility variants to psoriasis and downstream functional study to elucidate the underlying mechanisms of diseases are also needed. Taken together, unremitting efforts of the basic research on psoriasis will lead us to achieve a better treatment and diagnosis for psoriasis in the near future.

Aspherical optical elements are widely used in modern optic system design. The tool model and workpiece deformation are established, and the tool radius error compensation is discussed. The aspherical optical elements are processed on a precision milling machine, and tested on a measuring instrument.

To investigate the mechanisms of Alternanthera philoxeroides (Mardus) Grisebach in inhibiting Oncomelania snails' locomotivity and killing effect.Uninfected snails were divided into four groups and exposed to an aqueous extract of A. philoxeroides and dechlorinated water (as control) for 12 h or 20 h, respectively. The activities of the Mg2+-adenosine triphosphatase (Mg2+-ATPase), cholinesterase (ChE), lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) in the head-foot muscles, centric ganglions, gills and liver of Oncomelania hupensis were analyzed using enzyme histochemistry technology and changes were observed under a light microscope. Statistical quantitative analysis of data of grey values was conducted on the computer-assisted image analyzing system (HPIAS-1000).The color of stained ChE in the head-foot muscles, centric ganglions and gills of the snail lightened evidently, showing a decrease of ChE activity after snails were immersed in the extract of A. philoxeroides for 12 h or 20 h. Results of grey values at different stained parts of snail, measured by the computer-assisted image analyzing system, indicated that there was a significant difference (P0.01) between the activities of ChE in the head-foot muscles (130.95 +/- 8.08, 129.91 +/- 7.05), centric ganglions (127.43 +/- 7.27, 126.78 +/- 7.38) and gills (121.38 +/- 7.31, 126.41 +/- 8.28) from snails exposed to aqueous extract of A. philoxeroides for 12 h and 20 h and the activities of the enzyme in counter-parts (64.65 +/- 8.54, 65.18 +/- 7.96, 57.86 +/- 6.57, 50.71 +/- 6.15, 88.96 +/- 6.78 and 89.86 +/- 7.01, respectively) from control group. The activities of Mg2+-ATPase also showed a significant difference (P0.05) in the head-foot muscles (89.91 +/- 5.08), centric ganglions (71.15 +/- 5.43) and liver (112.40 +/- 7.81) of the snail after 20 h of exposure against those in counter-parts (78.81 +/- 8.10, 60.09 +/- 6.05 and 95.50 +/- 8.35, respectively) from the control, but no significant difference (P0.05) was shown on the activities of Mg2+-ATPase between the snails exposed for 12 h in extract of A. philoxeroides and those of control. No statistical difference (P0.05) was found between the dechlorinated water group and the extract of A. philoxeroides group in the activities of LDH and SDH after 12 h or 20 h of exposure.The extract of A. philoxeroides rapidly inhibits ChE, and then the activity of Mg2+-ATPase, which may suppress the release and utilization of ATP in the Oncomelania snails and finally causes death of snails.

Keloids are benign fibroproliferative dermal tumors of unknown etiology. Some studies have suggested that human HLA status might potentiate development of keloids phenotype. No report has been published about HLA class I alleles associated with keloids in Chinese Han individuals. To investigate the etiology of keloids, the polymerase chain reaction-sequence-specific primer (PCR-SSP) method was used to analyze the distribution of HLA class I alleles in 192 patients with keloids and 252 healthy controls in a Chinese Han population. The frequencies of HLA-A*03 (6.77% vs 0%, p(c)10(-7)), A*25 (10.16% vs 4.56%, p(c) = 0.0111), B*07 (7.81% vs 2.58%, p(c) = 0.0080), and Cw*0802 (19.79% vs 10.32%, p(c) = 0.0004) were significantly increased in keloid patients, whereas the frequency of HLA-A*01 (18.75% vs 38.10%, p(c)10(-7)) was highly decreased, compared with that in healthy controls. The A*03-B*07, A*25-B*07, A*03-Cw*0802, A*25-Cw*0802, and B*07-Cw*0802 were found as high-risk haplotypes in developing keloids in this study. No extended haplotype was found to be significantly related to keloids. Through stratified analysis, the association of subgroups (single site/multiple site, severity, and family history) of keloid patients with specific HLA alleles was identified. Our data suggest these alleles may be keloids susceptibility genes or may be in close linkage with the susceptibility genes.

Three clinical strains of Escherichia coli (p168, p517 and p667) were collected in 2006 from three hospitals in Anhui Province (China). PCR and DNA sequencing revealed that E. coli p168 carried a novel extended-spectrum β-lactamase (ESBL), which was designated CTX-M-87. The extended-spectrum β-lactamase which was carried by E. coli p517 and E. coli p667 was previously named CTX-M-65. The deduced amino acid sequence of CTX-M-87, with pI 9.1, differed from that of CTX-M-14 by the substitutions Ala77→Val and Pro167→Leu. Like CTX-M-14, CTX-M-87 had a more potent hydrolytic activity against cefotaxime than against ceftazidime and had high affinity for cefuroxime and cefotaxime. These data show that mutations at position 167 in CTX-M do not always affect catalytic activity and substrate preference.

In Computer Controlled Optical Surfacing (CCOS), polishing tool-path is the base of solving other control parameters such as dwell time. In order to improve the fabrication results of polishing off-axis aspheric, a novel method to optimize the tool-path is discussed in this paper. The optimizing method named weighted-iterative algorithm is according to the balance principle of the particle system. The power factor of each dwell point represents the requirement of dwell density. Considering the factors which influence the polishing result, the power factors cosist of three elements include constant, error distribution and dwell distance of workpiece edge. The tool-path is solved by numerical iterative method. In the end, an error data is simulated with actual parameters using the matrix-based algorithm with two different tool-paths. The one is X-Y uniform spacing model and the other one is to optimize it based on the first. The comparison shows that the results of the optimized one are much better than traditional one, especially the rms convergence rate. Theory of the algorithm is simple and exercisable, and it satisfies practical requirement as well.

SiC is a preferential material for space large aperture mirror at present. Aperture of 1 meter sic mirror can meet most requirements of space optical systems. For larger aperture sic mirror, it is not economical and safe to make the monolithic body directly. The SiC mirror brazed assembly with several segments can solve these problems. In this paper, one Φ600 mm sic plane mirror as demonstration welded with 3 radial segments is roughly grinded to the required figure at first, then the mirror is finely grinded and polished by CCOS technology, the RMS of the figure error is less than 1/20λ (λ = 632.8nm) finally. The problems of the preferential removal near braze joint and material removing rate influenced by deformation of mirror surface are solved. Methods of reducing the residual stress of mirror are discussed. At last, some measurements and experiments are carried on, and the results conclude that the brazed SiC mirror can meet the requirement of large aperture optical systems.

A brand-new technology for manufacturing SiC reflecting mirror that is different from the traditional method is adopted, which is called as fixed abrasive surfacing technology. Not only the new method achieves better mirror quality with bigger diamond in diameter quickly , but also because of the fixed motion between the abrasive and workpiece and it will be good for surface finishing. During experiment, material removal characteristic which is on SiC under certain rotation speed and pressure by W7,W5,W3.5,W1.5 pellets has been tested. Through those removal curves, we come to a conclusion that the technology not only has a higher removal rate, but also has much more stability. In addition, the surface roughness experiment is mentioned. In the first stage, we achieved a mirror with surface roughness 42.758nm rms with W7 pellets. The surface roughness is descending as we change pellets with smaller diamond in diameter . At the end of experiment, a smooth surface with roughness 1.591nm rms has been achieved after using W1.5 pellets. Experiment results indicate that the technology which manufactures SiC reflecting mirror with fixed abrasive is able to replace the traditional slurry abrasive completely in certain finishing phase and also has a great foreground in application.

The aim of this study was to investigate the regulation of 5-aza-CdR on transcription of SHP-1 gene and effects on the proliferation and apoptosis of K562 cells. Methylation-specific PCR (MSP) was used to detect CpG island methylation in SHP-1 promoter. MTT and flow cytometry were used to detect the growth and apoptosis of K562 cells after treatment with different concentration of 5-aza-CdR. The expressions of SHP-1 mRNA and protein were determined by FQ-PCR and Western blot. The expression of p-JAK2 was assayed by Western blot. The result showed that methylation of SHP-1 gene promoter was detected in K562 cells, and the SHP-1 mRNA and protein were expressed again in K562 cells after treatment with 5-aza-CdR, meanwhile the expression of phosphorylated P-JAK2 was down-regulated; 5-aza-CdR significantly inhibited the cell growth in dose and time dependent manners. AG490 inhibited the cell proliferation. 5-aza-CdR increased the apoptosis rate of K562 cells also in dose- and time-dependent manners. The apoptosis rates of K562 cells treated with 5-aza-CdR for 1, 3 and 5 days were 9.3%, 24.2% and 37.7% respectively. After treatment with 2 micromol/L 5-aza-CdR for 24 hours, cells in G(0)/G(1) phase increased gradually, cells in G(2)/M phase decreased gradually, cells were arrested in G(0)/G(1) phase. The cell ratios in G(2)/M phase at 1, 3 and 5 days after treatment with 5-aza-CdR were 30.7%, 23.45 and 19.3% respectively. It is concluded that the 5-aza-CdR, inhibitor of specific methylation transferase, can re-express the silent SHP-1 gene in K562 cells, inhibits the proliferation of leukemia cells and induces cell apoptosis by activating JAK/STAT pathway.

To observe effects of different interference orders of electroacupuncture and exercise therapy on the therapeutic effect of hemiplegia after stroke.The patients of hemiplegia due to stroke were randomly divided into a group A and a group B. The group A were treated by exercise therapy after electroacupuncture (EA) and the group B by EA after exercise therapy. Fugl-Meyer evaluation (FME), modified Barthel index (MBI) and amplitude of somatosensory evoked potential (SEP) on the affected side before treatment and after one therapeutic course (2 weeks) were recorded and compared in the two groups.Before and after treatment, FME were 31.760 +/- 4.438 and 76.640 +/- 3.621, and MBI were 26.520 +/- 2.413 and 62.360 +/- 3.700 in the group A; and FME were 32.480 +/- 5.903 and 65.640 +/- 5.212, and the MBI were 28.000 +/- 3.383 and 54.480 +/- 5.205 in the group B, respectively, with very significant differences in FME and MBI in the two groups (all P0.01); and the different values before and after treatment in the two indexes in the group A were better than those in the group B (all P0.05). There was a very significant difference in the amplitude of SEP before and after treatment in the group A (P0.01) and no significant difference in the group B, and there was no significant difference between the two groups in the different value of the amplitude of SEP.Combination of any orders of electroacupuncture and exercise therapy can improve limb function of the patient with hemiplegia after stroke, but the therapeutic effect of exercise therapy after EA is better.

This study was aimed to investigate the effect of recombinant mutant human TNF-related apoptosis-inducing ligand (rmhTRAIL) combined with As(2)O(3) on inducing apoptosis of adriamycin-resistant leukemia cell line K562/A02 (mdr-1(+)). The morphologic changes of cells treated with rmhTRAIL were observed by inverted microscope, taking adriamycin-sensitive cell line K562 (mdr-1(-)) as control; the inhibitory rate of cell proliferation after being treated with rmhTRAIL, As(2)O(3) alone or combined was assayed by MTT method; the apoptosis peaks of K562/AO2 and K562 were quantitatively detected by flow cytometry with PI staining after being treated with rmhTRAIL, As(2)O(3) alone or in combination. The results indicated that the inhibition effect of rmhTRAIL and As(2)O(3) in combination on K562/AO2 and K562 cells was higher than that of riTRAIL and As(2)O(3) alone (p0.01), rmhTRAIL combined with As(2)O(3) had synergistic effect in killing K562/AO2 and K562 cells by king's formula. The apoptosis rates of K562/AO2 and K562 cells were 34.93 +/- 0.10% and 10.53 +/- 0.16% (p0.01), as well as 5.95 +/- 0.07%, and 3.50 +/- 0.01% (p0.05), 50.95 +/- 0.91% and 20.75 +/- 0.95% (p0.05) respectively when their cells were treated by rmhTRAIL and As(2)O(3) alone. The apoptosis rate in K562/AO2 group was higher than that in K562 group. It is concluded that rmhTRAIL can induce K562/A02 and K562 cell apoptosis; rmhTRAIL combined As(2)O(3) had synergistic effects; the efficacy of on rmhTRAIL or As(2)O(3) inducing K562/AO2 cell apoptosis is higher than that on their parental cell line K562.

We developed an oligonucleotide biochip for synchronous multiplex detection of 31 known mitochondrial DNA mutations associated with MELAS (Mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes) and MERRF (Myoclonic epilepsy with ragged red fibers). Allele-specific oligonucleotide probes were covalently immobilized on aldehyde modified glass slides, and then hybridized with Cy5-labled DNA fragments amplified from sample DNAs by a multiplex asymmetric PCR (MAP) method. Five patients with MELAS, 5 patients with MERRF and 20 healthy controls were investigated using the oligonucleotide biochip. The results showed that all the cases with MELAS had an A3243G mutation in the MT-TL1 gene. In the MERRF group, 4 cases were found to be an A8344G mutation and 1 case was a T8356C mutation, and both mutations were in the MT-TK gene. In the healthy controls, none of the 31 related mutations was found. The results of the DNA biochip were consistent with those by DNA sequencing. Clearly, the DNA biochip combined with MAP method would become a valuable tool in multiplex detecting of the point mutations in mtDNA leading to MELAS and/or MERRF syndrome. Moreover, this biochip format could be modified to extend to the screening scope of SNPs for any other human mitochondrial diseases.

To investigate the expression and mutation of Mad1, Mxi1 and Rox genes in leukemia cells.Expression and mutation of Mad1, Mxi1 and Rox genes in bone marrow mononuclear cells (BMMNC) from 26 de novo acute leukemia (AL) patients, and in peripheral blood mononuclear cells (PBMNC) from 30 healthy volunteers, as well as in 7 human leukemic cell lines were analyzed by reverse transcription-polymerase chain reaction (RT-PCR), single strand conformational polymorphism (SSCP) and DNA sequencing.RT-PCR showed that all the above cells expressed Mad1, Mxi1 and Rox mRNA. SSCP revealed four polymorphisms: two in Mad1, one each in Mxi1 and Rox. DNA sequencing detected nine missense mutations: two in Mad1 in AL patients, four in Mxi1 (three in AL patients and one in KG-1 cell line), and three in Rox in AL patients. The mutations of Mad1, Mxi1 and Rox mRNA were detected in 2, 3 and 3 patients, respectively.It is for the first time to demonstrate the mutations of Mad1, Mxi1 and Rox genes in AL patients suggesting these mutated genes involve in the pathogenesis of leukemia.

The aim of this work was to study the phenotypic and molecular characterization of a novel plasmid-mediated AmpC β-lactamase from Escherichia coli E384. Conjugation experiments, isoelectric focusing, pulsed-field gel electrophoresis, plasmid profiling, and Southern blot as well as PCR, sequencing techniques, and susceptibility testing were carried out to investigate the underlying mechanism of resistance. The kinetic parameters were determined to characterize the novel enzyme. MIR-4 β-lactamase, pI 8.2, is a novel variant with four substitutions of amino acids compared with the sequence of MIR-1. E. coli E384 displays resistance to eight β-lactam antimicrobial agents and three fluoroquinolones. The minimal inhibitory concentrations of β-lactam in combination with β-lactamase inhibitors show no significant synergy. Kinetic parameters suggest that the novel enzyme effectively hydrolyzes broad-spectrum β-lactams. The same hybridization signal was detectable only in the 54-kb plasmid band that hybridized with the bla CTX-M- and bla ampC-specific probes. This is the first description of a plasmid-mediated MIR-4 enzyme in China. This study illustrates the importance of molecular surveillance in tracking AmpC-producing strains at general hospitals and emphasizes the need for epidemiological monitoring.

Two clinical strains of Klebsiella pneumoniae (K. pneumoniae) and one isolate of Escherichia coli (E. coli) were collected from two large general hospitals in China. Conjugation experiment, susceptibility testing, isoelectric focusing, PCR, and sequencing techniques as well as clone, expression, purification and kinetics were carried out to describe the characterization of the novel SHV-tpye enzyme. The analysis of plasmid profiling and pulsed-field gel electrophoresis of the novel enzyme were performed to investigate epidemiology. These isolates had CTX-M-14 and SHV-89 beta-lactamases. SHV-89 beta-lactamase of pI 7.6 is a novel variant with two substitutions compared with the sequence of SHV-1: Leu35Gln and Met129Val. Its gene also had two silent mutations at positions 369 and 774, respectively. The results of substrate profiles and MIC determinations showed the activity of the novel enzyme was insufficient for the enzyme to count as an extended-spectrum beta-lactamase (ESBL). The substrates of the enzyme were also characterized. Furthermore, the three novel SHV enzyme-producing strains were epidemiologically unrelated. The emergence of a novel SHV-type beta-lactamase is rarely described in other areas. This study illustrates the importance of molecular survelliance in tracking SHV-producing strains in large teaching hospitals and emphasizes the need for epidemiological monitoring.

Trichloroethylene (TCE), a colorless and volatile organic solvent, has long been a major chemical hazard during occupational exposure because of its extensive use in industry. Exposure to TCE can cause significant skin lesions, but the effect of TCE on skin irritation has received little attention. We therefore investigated the effect of TCE on skin irritation and oxidative stress using hairless mice. BALB/c hairless mice were subjected to acute and cumulative topical TCE treatment. Skin reactions were evaluated by visual inspection, histopathology examined by microscopy and oxidative stress assessed by measurement of malondialdehyde (MDA) levels, superoxide dismutase (SOD) activities and nitric oxide (NO) production. Under acute and cumulative TCE irritation, the skin developed erythema and edema, and the predominant histopathological features were hyperkeratosis, spongiosis and inflammatory cell infiltrates. In parallel to these morphological changes, acute TCE irritation also concentration-dependently increased MDA levels and inhibited SOD activities of the skin. However, in cumulative irritation, the MDA levels and SOD activities were initially elevated with increased TCE concentrations, but thereafter reduced with further concentration increments; the linear dose-response relationship was not preserved. TCE also concentration-dependently increased NO production both in acute and cumulative irritation. These results suggest that TCE is capable of producing skin irritation effect in vivo, with histopathological changes characterized by hyperkeratosis, spongiosis and inflammatory cell infiltrates. Moreover, oxidative stress may be associated with the clinical manifestations and histopathological abnormalities in TCE-induced skin irritation.

Hydrothermal synthesis of borosilicate zeolites with BEA structure and the modification of HB-Beta zeolites with orthophosphoric acid were performed. The samples were characterized by the X-ray diffraction, FT-IR, nitrogen adsorption, chemical analysis, 11 B and 31 P MAS NMR technique. The effects of the phosphorus-modification on the structure, crystallinity, acidity, framework vibrations and textural properties of the HB-Beta were investigated systematically. The coordination environments of boron and phosphorus in the framework of P-HB-Beta zeolites were revealed. The formation of P(BO) 4-n (SiO) n -like species is observed.

The WBN/Kob- Lepr fa rat is a new congenic strain for the fa allele of the leptin receptor gene ( Lepr ). Homozygous ( fa / fa ) WBN/Kob- Lepr fa rats provide a model of non-insulin-dependent diabetes with obesity. Here, we describe the characteristics of this new animal model in detail. At 7 weeks of age, both male and female obese WBN/Kob rats showed inflammatory cell infiltration of the pancreas that suggested pan-pancreatitis and an abnormal OGTT. At 3 months of age, both male and female obese WBN/Kob rats developed overt diabetes mellitus associated with severe chronic pancreatitis. In contrast, lean female WBN/Kob rats do not develop pancreatitis or diabetes. In WBN/Kob rats, this mutation might promote the onset of severe pancreatitis, leading to the rapid development of diabetes mellitus.

To record and analyze the characteristics of condyle movement before and after functional treatment in Angle class II division 1 malocclusion (II(1)) patients.Twenty Angle II(1) patients were included in the study and 20 Angle I patients without temporomandibular disorders symptoms served as controls. Computer aided diagnosis axiograph (CADIAX) was used to record the condyle movement in Angle II(1) patients and that caused by functional appliance treatment during mouth opening and closing.The tracing were vertically displayed. Value Z [L: 2.21 (1.01) mm, R: 1.12 (1.02) mm] and sagittal condylar inclination [L: 12.89 (9.12) degrees, R: 12.12 (8.89) degrees] were increased significantly in Angle II(1) patients compared with those in control group (P0.05). After functional treatment, the condyle movement in Angle II(1) patients was close to normal (P0.05).Typical characteristics of condyle movement were found in Angle II(1) patients and it was stable and repeatable. Functional therapy had a positive effect on temporomandibular joint function in Angle II(1) patients.

Angioimmunoblastic T-cell lymphoma (AILT) is a peripheral T-cell lymphoma often complicated autoimmune phenomena such as autoimmune cytopenia, and is a truly rare type of NHL. In order to investigate the clinical features, pathological manifestation of this lymphoma, and to explore its therapy protocol, a 37-years old patient with AILT was investigated. The routine blood examination, bone marrow smear, lymphonodus biopsy, Coombs test, flow cytometry for bone marrow mononuclear cells, serological test, immunochemistry method etc were performed for this patient. The results showed that the systemic lymphadenectasis and hepatosplenomegaly were seen in patient, the cervical lymphonode biopsy revealed AITL. The hematoglobin level and number of reticulocytes were very low. Coombs test was positive. Simultaneously, the bone marrow aspirate revealed erythroid aplasia. The warm type autoimmune hemolytic anemia (AIHA) and pure red cell aplasia (PRCA) were co-existed. After one course of chemotherapy with CHOP-E, infiltration sign of AITL patient with AIHA and PRCA disappeared. In conclusion, the AITL patient complicated with AIHA and PRCA was successfully diagnosed, the lymphonode biopsy and bone marrow smear showed more significant, the chemotherapy protocol of CHOP-E can give some effect to cure such angioimmunoblastic T cell lymphoma.

To improve the recognition of immunoglobulin D multiple myeloma and explore its clinical feature and laboratory examination characteristics, so as to reduce the the missed diagnosis and misdiagnosis, a case of IgD multiple myeloma (MM) with myelofibrosis and bone marrow necrosis is reported. The clinical feature, treatment and prognosis of IgDlambda MM were discussed. Immunoglobulin D multiple myeloma is a rare disease and predominantly occurs in young male patients, which shows an aggressive clinical course with poor response to conventional treatment and unfavorable prognosis. Immunoglobulin D multiple myeloma was usually misdiagnosed as a light chain type multiple myeloma by using routine laboratory examination. Immunoglobulin D monoclonal protein is not easy to be detected owing to its low protein level, resulting in missed diagnosis. Immunofixation electrophoresis is highly sensitive and specific for diagnosis of IgD MM, can enhance accuracy of diagnosis for this rare disease.

Estrogens have been associated with risk for epithelial ovarian cancer (OVCA). Both IL-6 and IL-8 are also likely involved in the progression of OVCA. In order to discover the underline molecular mechanism, we investigated the modulation of estrogen and two cytokines in the growth and progression of epithelial OVCA. In these studies, the effect of 17beta-estradiol (E(2)) on the expression levels of IL-6, IL-8 and their receptors was investigated. The effect of IL-6 and IL-8 on activation of estrogen-responsive promoter as well as estrogen receptor (ER)alpha and ER beta expression was also analyzed. Gene expression profile analysis revealed that CAOV-3 and OVCAR-3 cells, which express ER, IL-6 and IL-8 receptors, are suitable model for this study. We found that E(2) not only enhanced IL-6 and IL-8 production via NF-kappaB signaling pathway, but also modulated their respective receptor expression. Tamoxifen (Txf), an ER antagonist, completely abolished E(2)-stimulated cell growth and the expression of IL-6 and IL-8. IL-6/IL-8-induced cell proliferation was completely blocked by their specific neutralizing antibodies, which partially inhibited E(2)-induced cell growth. In the absence of estrogen, both cytokines activated estrogen-responsive promoter, which was completely blocked by Txf, and caused a dose-dependent ER alpha increase and ER beta decrease. Pretreatment of OVCAR-3 with p38 MAPK, MEK1/2 or ErbB2 MAPK inhibitors, respectively, blocked IL-6-mediated induction of estrogen-responsive promoter while Src inhibitor blocked IL-8-induced activation of estrogen-responsive promoter. These results provide a novel mechanism that estrogens, IL-6 and IL-8 may form a common amplifying signaling cascade to modulate OVCA growth and progression. Estrogen-induced OVCA proliferation is partially occurring via enhanced IL-6 and IL-8 production and modulated their receptors, and IL-6/IL-8 could also promote OVCA growth through an ER alpha pathway.

To analyze the mutation of TSC gene in two sporadic patients with tuberous sclerosis complex (TSC).All the coding exons of TSC1 and TSC2 genes of these two patients, unaffected member in the two families, and 100 unrelated population-matched controls were amplified by polymerase chain reaction. The products were analyzed by direct sequencing.Two TSC2 gene mutations (c. 268CT, c. 5 227CT) were identified in two patients, but not in their family members and in 100 unrelated population-matched controls.These two mutations are the cause of the clinical phenotypes of these two sporadic patients with TSC.

To study a Chinese pedigree with Hailey-Hailey disease (HHD) and examine the ATP2C1 gene mutation in this family.All exons of ATP2C1 gene were analyzed with polymerase chain reaction and DNA sequencing in all patients of this family and 100 unrelated population-match controls.We identified a novel heterozygous nucleotide A --G transition at position 235 - 2 in intron 3 of ATP2C1 gene. This splice site mutation was not found in the healthy members of this pedigree and in the controls.The splicing mutation can affect the result of transcription and translation, and it is a specific novel mutation of ATP2C1 gene.

To identify the mutations of ED1 gene in a family with X-linked hypohidrotic ectodermal dysplasiaEight coding exons of ED1 gene of two patients with clinically confirmed X-linked hypohidrotic ectodermal dysplasia, their parents, and 100 unrelated population-matched control were amplified by polymerase chain reaction. The products were further analyzed by direct sequencing.Two patients with X-linked hypohidrotic ectodermal dysplasia in this pedigree showed a point mutation at nucleotide 1 045 ( AG) . Meanwhile, heterozygous double peaks of nucleotide G and A at the same position were found in their mother, but not in their father and 100 unrelated population-matched controls.The c. 1 045AG mutation of ED1 gene may be the pathologic cause of this Chinese family with X-linked hypohidrotic ectodermal dysplasia.

Dermatovenereology have developed rapidly in fields including genodermatology, dermatological epidemiology, skin immunology, infectious skin diseases, clinical diagnosis, drug treatment, light therapy, and sexually transmitted diseases in recent years. Meanwhile, many skin diseases and a few venereal diseases still lack effective ways of treatment and control. More efforts should be made in both basic and clinical researches in the related fields.

The aim was to construct a prokaryotic expression plasmid encoding the fusion gene of single-chain variable fragment and monocyte chemotactic protein-3 (MCP-3). The cDNAs of immunoglobulin (Ig) VH and Ig VL were amplified by RT-PCR and assembled into the single-chain variable fragment (scFv) by recombinant PCR method. The cDNAs of Ig VH and Ig VL were connected by a (Gly4Ser)3 linker. Then, the fragments of scFv and MCP-3 were connected with a NDAQAPKS spacer, using recombinant PCR method again. The results indicated that the fusion gene of scFv-MCP-3 were constructed correctly and cloned into the prokaryotic expression plasmid successfully identified by sequencing and restriction endonucleases examination. Finally, the fusion protein was expressed in E coli DH5alpha under induction by arabinose. And the fusion protein was 65 kD and account for 30% of the total protein of the bacteria. In conclusion, a prokaryotic plasmid, encoding the fusion gene of single-chain variable fragment with MCP-3 and expressing idiotype protein vaccination against B cell lymphoma, was constructed correctly.

The aim of study was to investigate the combined effect of recombinant mutant human TRAIL (rmhTRAIL) with daunorubicin (DNR) or alone on K562 and U937 leukemia cell lines and its mechanism. The fibroblasts (MRC-5) of normal-human embryonic lung were used as control cells. After being treated with rmhTRAIL and DNR or only with rmTRAIL, the cytotoxic effect and the apoptosis rate in K562, U937 cells were measured by MTT assay. The expression levels of TRAIL death receptor and TRAIL decoy receptor mRNA in these three cell lines were assayed by semiquantitive RT-PCR before and after treatment with DNR. The results indicated that K562 and U937 were sensitive to rmhTRIAL. DNR had synergistic inhibitory effect with rmhTRAIL on the growth of K562 and U937 cell lines (P0.05). The expression level of DR4 and DR5 mRNA was significantly higher in K562 and U937 with combined treatment of rmhTRAIL and DNR than that in those alone, while the expressions of DcR1 and DcR2 mRNA were not influenced. It is concluded that in vitro, rmhTRAIL alone or in combination with DNR can obviously inhibit the growth of leukemia cell lines and induce cell apoptosis, DNR and rmhTRAIL have a synergistic inhibitory effect on growth of K562 and U937. The mechanism may correlate with the up-regulation of DR4 and DR5 of K562 and U937.

To study the effects of IL-13 on the differentiation and expression of transcription factor c-fos of human erythroleukemia cell line (HEL) cells.Reverse transcription polymerase chain reaction (RT-PCR) was used to observe the mRNA expression of IL-13 receptor a 1, GP i b, vWF and c-fos, and Western blot and cytometry were used to analyse their protein expression.IL-13 receptor a 1 was expressed on HEL cells. IL-13 (100 ng/ml ) up-regulated the mRNA expression of GP II b and vWF. The ratio of luminous absorption (LA) of GP I b to p-actin bands ( AB) was 1. 303 in control group, whereas was 2. 912 in experiment group; being 2. 23-fold higher than that in control group (P0. 05). The ratio of LA to AB for vWF was 0.217 in control group, and 0. 506 in experiment group; indicating a 2. 33-fold increase in experiment group (P0. 05). The protein expression of GP I b and vWF was significantly increased in experiment group, compared with that in control group. IL-13 inducing the increased expression of c-fos mRNA and protein of HEL cells peaked at 30 min and 60 min, respectively. The ratio of LA to AB for c-fos was also increased at 30 min and 60 min (P0. 05).IL-13 prompts the differentiation of HEL cells and up-regulates the expression of c-fos.