Correlation between serum uric acid levels and carotid plaque neovascularisation assessed by contrast-enhanced ultrasound.

AIM: To investigate the potential impact of serum uric acid (SUA) levels on the grade of intraplaque neovascularisation (IPN) at contrast-enhanced ultrasound (CEUS). MATERIAL AND METHODS: The study screened 212 patients with carotid atherosclerotic plaques using conventional ultrasound, and the patients then underwent CEUS. Based on the distribution of contrast medium microbubbles in the plaque, patients were split into three groups: 60 patients regarded as grade 0-1 (group A), 81 patients as grade 2 (group B), and 71 patients as grade 3 (group C), and SUA levels were measured on the second day after CEUS. RESULTS: The frequencies of stroke were statistically different between the three groups (p<0.05). In grades 0, 1, 2, and 3, SUA levels ranged from 236.92 ± 72.75, 276.46 ± 67.31, 283.93 ± 53.85, and 384.49 ± 79.80 µmol/l, respectively. Spearman's analysis showed that the visual grade of IPN at CEUS correlated linearly with the SUA level (r=0.551, p<0.01). The difference in SUA levels between different sexes was statistically significant (p<0.05), and the differences in plaque echogenicity and diastolic blood pressure (DBP) between the three groups were also statistically significant (all p<0.05). CONCLUSIONS: SUA levels correlate positively with the visual grade of IPN at CEUS, which may promote plaque vulnerability. The present results may further help to optimise therapy for vulnerable plaque and improve stroke risk stratification strategies.

Safety and effectiveness of combined scoring balloon and paclitaxel-coated balloon angioplasty for stenosis in the hemodialysis access circuit.

PURPOSE: To evaluate the safety and effectiveness of combined scoring balloon (SB) and paclitaxel-coated balloon (PCB) angioplasty for stenosis in the dysfunctional hemodialysis access circuit. MATERIAL AND METHODS: Patients were referred from outpatient dialysis centers by their nephrologists because of dysfunctional dialysis access circuit. Fistulogram/graftogram was performed by experienced interventional radiologists. Those with in-stent stenosis, stent edge stenosis or vessel diameter at the culprit segment larger than 6mm were excluded. Angioplasty of the stenotic segment was performed with SB and followed by PCB. All study outcomes were defined according to the Society of Interventional Radiology technology assessment committee reporting standards for percutaneous interventional procedures in dialysis access circuit. RESULTS: A total of 23 patients received combined SB/PCB angioplasty for stenosis of hemodialysis access circuit which included 15 fistulas and 8 grafts. There were 10 men and 13 women with a mean age of 63.3±2.7 (SD) years (range: 37-85years). The technical success and clinical success rates were both 100%. There were no complications during or after the procedures. The target lesion primary patency rates at 3, 6 and 12months were 91.3%, 69.6%, and 45.2%, respectively and the estimated median target lesion restenosis (TLR) free duration was 11.0months (95% confidence interval [CI]: 5.1-16.9months). In patients with a recurrent stenosis, the median TLR-free duration of combined angioplasty was significantly higher than that of prior angioplasty with plain balloon (10.2months [95% CI: 6.4-14.0months] vs. 4.2months [95%CI: 2.1-6.4months]) (P=0.047). The mean TLR-free duration was significantly higher in patients with a juxta-anastomotic stenosis than those with non-juxta-anastomotic lesion (21.3months [95% CI: 14.7-28.0months] vs. 8.2months [95% CI: 5.1-11.4months]) (P=0.004). CONCLUSION: Combined SB/PCB angioplasty is safe and effective for the hemodialysis access stenosis.

Endometrial carcinoma: diffusion-weighted imaging diagnostic accuracy and correlation with Ki-67 expression.

AIM: To evaluate the role of diffusion-weighted imaging (DWI) in the detection of endometrial carcinoma and to correlate the apparent diffusion coefficient (ADC) value with Ki-67 expression. MATERIALS AND METHODS: Fifty-two patients with invasive cancer who underwent pelvic MRI were prospectively evaluated using DWI with b-values of 0 and 1000 s/mm2.The ADC values from standard DWI were measured. The expression of Ki-67 in histological specimens was analysed using immunohistochemistry. The ADC values of endometrial carcinoma and normal endometrial parenchyma were compared. Relationships between ADC values and Ki-67 expression were determined using Wilcoxon's signed rank test and the Kruskal-Wallis test. RESULTS: Endometrial carcinoma was detected at DWI as a hyperintense area in 92.3% (48/52) of patients. There was a significant difference in the mean ADC values between endometrial carcinoma and normal endometrial parenchyma (1.39±0.27×10-3 versus 0.93±0.21×10-3 mm2/s, p<0.001). The mean ADC values of grade 1 patients were significantly higher than those of grade 3 patients (1.01±0.16×10-3 versus 0.83±0.21×10-3 mm2/s, p<0.05). The mean ADC values of stage IB patients were significantly lower than those of stage IA patients (0.86±0.16×10-3 versus 1.04±0.21×10-3 mm2/s, p<0.01). The mean ADC values of high Ki-67 expression patients were significantly lower than those of low Ki-67 expression patients (0.82±0.12×10-3 versus 1.16±0.12×10-3 mm2/s, p<0.001). There was a significant negative correlation between the mean ADC value and Ki-67 expression (r=-0.82, p<0.001). CONCLUSION: The ADC value was a helpful parameter for detecting the tumour grade, stage, and proliferation of endometrial carcinoma, and may further improve patient prognosis and contribute to the development of more effective treatment programmes.

Outcomes of the six-strand M-Tang repair for zone 2 primary flexor tendon repair in 54 fingers.

We repaired complete divisions of flexor tendons in zone 2 in 54 fingers using a six-strand core M-Tang repair method. Partial active digital motion started with early passive digital motion carried out first in the first 3-4 weeks after surgery and full range of active motion in later weeks. The patients were followed for 4-27 months. According to Strickland criteria or Tang criteria, 24 (83%) had excellent or good, four fair and one poor results in 28 fingers with follow-up of more than one year. In the other 25 fingers which were followed for less than 12 months, 19 (76%) had excellent and good, four fair and two poor results. There were no repair ruptures. We analysed outcomes against ages, gender, pulley integrity, accompanied injuries and follow-up times. The patients younger than 37 years old, male patients and with their A2 pulley(s) vented there were significantly better outcomes. The patients with longer than one year follow-up had significantly smaller extension deficits than those with less than one year follow-up. LEVEL OF EVIDENCE: IV.

Flow cytometry and K-mer analysis estimates of the genome sizes of Bemisia tabaci B and Q (Hemiptera: Aleyrodidae).

The genome sizes of the B- and Q-types of the whitefly Bemisia tabaci (Gennnadius) were estimated using flow cytometry (Drosophila melanogaster as the DNA reference standard and propidium iodide (PI) as the fluorochrome) and k-mer analysis. For flow cytometry, the mean nuclear DNA content was 0.686 pg for B-type males, 1.392 pg for B-type females, 0.680 pg for Q-type males, and 1.306 pg for Q-type females. Based on the relationship between DNA content and genome size (1 pg DNA = 980 Mbp), the haploid genome size of B. tabaci ranged from 640 to 682 Mbp. For k-mer analysis, genome size of B-type by two methods were consistent highly, but the k-mer depth distribution graph of Q-type was not enough perfect and the genome size was estimated about 60 M larger than its flow cytometry result. These results corroborate previous reports of genome size based on karyotype analysis and chromosome counting. However, these estimates differ from previous flow cytometry estimates, probably because of differences in the DNA reference standard and dyeing time, which were superior in the current study. For Q-type genome size difference by two method, some discussion were also stated, and all these results represent a useful foundation for B. tabaci genomics research.

Highly Selective and Potent Thiophenes as PI3K Inhibitors with Oral Antitumor Activity.

Highly selective PI3K inhibitors with subnanomolar PI3Kα potency and greater than 7000-fold selectivity against mTOR kinase were discovered through structure-based drug design (SBDD). These tetra-substituted thiophenes were also demonstrated to have good in vitro cellular potency and good in vivo oral antitumor activity in a mouse PI3K driven NCI-H1975 xenograft tumor model. Compounds with the desired human PK predictions and good in vitro ADMET properties were also identified. In this communication, we describe the rationale behind the installation of a critical triazole moiety to maintain the intricate H-bonding network within the PI3K receptor leading to both better potency and selectivity. Furthermore, optimization of the C-4 phenyl group was exploited to maximize the compounds mTOR selectivity.

Pathways contributing to development of spontaneous mammary tumors in BALB/c-Trp53+/- mice.

Mutation and loss of function in p53 are common features among human breast cancers. Here we use BALB/c-Trp53+/- mice as a model to examine the sequence of events leading to mammary tumors. Mammary gland proliferation rates were similar in both BALB/c-Trp53+/- mice and wild-type controls. In addition, sporadic mammary hyperplasias were rare in BALB/c-Trp53+/- mice and not detectably different from those of wild-type controls. Among the 28 mammary tumors collected from BALB/c-Trp53+/- mice, loss of heterozygosity for Trp53 was detected in more than 90% of invasive mammary tumors. Transplantation of Trp53+/- ductal hyperplasias also indicated an association between loss of the wild-type allele of Trp53 and progression to invasive carcinomas. Therefore, loss of p53 function seems to be a rate-limiting step in progression. Moreover, expression of biomarkers such as estrogen receptor alpha, progesterone receptor, Her2/Neu, and activated Notch1 varied among mammary tumors, suggesting that multiple oncogenic lesions collaborate with loss of p53 function. Expression of biomarkers was retained when tumor fragments were transplanted to syngeneic hosts. Tumors expressing solely luminal or basal keratins were also observed (27 and 11%, respectively), but the largest class of tumors expressed both luminal and basal keratins (62%). Overall, this panel of transplantable tumors provides a resource for detailed evaluation of the cell lineages undergoing transformation and preclinical testing of therapeutic agents targeting a variety of oncogenic pathways including cancer stem cells.

Effects of ischemic postconditioning on reperfusion injury in rat liver grafts after orthotopic liver transplantation.

AIM: The effects of ischemic postconditioning (IPostC) on ischemia reperfusion (IR) injury of liver grafts was examined in rats after orthotopic liver transplantation (OLT). METHODS: Male Wistar rats were used as donors and recipients to establish a liver transplantation model. The animals were randomly divided into four groups: sham-operated (SO, n = 6), IR (n = 6), IPostC1 (n = 6) and IPostC2 (n = 6). IPostC was achieved by several intermittent interruptions of blood flow in the early phase of reperfusion. Several parameters of hepatic damage, oxidative stress, neutrophil infiltration and the expression of TNF-alpha and MIP-2 were detected as well as microscopic examination. Nitric oxide release and liver NO synthases (endothelial NO synthase and inducible NO synthase) expression were also measured. RESULTS: We observed that a significant reduction in alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase values in two IPostC groups when compared with IR group. The increases in hepatic malondialdehyde, and decreases in superoxide dismutase and reduced glutathione levels after orthotopic liver transplantation were significantly inhibited by IPostC. IR induced increase in hepatic myeloperoxidase content, TNF-alpha and MIP-2 expression were also lowered by IPostC. The increases in NO content and NOS protein expression were much more prominent in IPostC treated groups. Animals treated with IPostC presented minimal hemorrhage and reduced signs of liver injury. There was no significant difference between two IPostC treated groups. CONCLUSIONS: IPostC provided significant protection against IR injury to liver grafts. The protective effect of IPostC is closely related to the NO production following the increase in endothelial and inducible NO synthases expression and the suppression of tumor necrosis factor-alpha and macrophage inflammatory protein-2 overproduction.

[Study on steroids of Cacalia tangutica].

Seven steroids and two cumarins were isolated from the petroleum ether extract of the specie Cacalia tangutica of the family Compositae which were collected in Minhe county, Qinhai province of China. The structures were identified as Stigmast4-en-3beta, 6beta-diol (1), 24-ethyl-5alpha-cholestane-3beta, 5, 6beta-triol (2), 7beta- methoxy-stigmast-5-en-3beta-ol (3), Schleicherastatin 1 (4), Stigmast-5-en-3beta, 7alpha-diol (5), umbelliferone (6) and hydrangetin (7) by the means of chemical and modern spectroscopic analysis (MS, 1H-NMR, 13C-NMR, DEPT). The compounds 1-5 were isolated from Cacalia tangutica for the first time.

Targeting FGF19 inhibits tumor growth in colon cancer xenograft and FGF19 transgenic hepatocellular carcinoma models.

Although fibroblast growth factor 19 (FGF19) can promote liver carcinogenesis in mice its involvement in human cancer is not well characterized. Here we report that FGF19 and its cognate receptor FGF receptor 4 (FGFR4) are coexpressed in primary human liver, lung and colon tumors and in a subset of human colon cancer cell lines. To test the importance of FGF19 for tumor growth, we developed an anti-FGF19 monoclonal antibody that selectively blocks the interaction of FGF19 with FGFR4. This antibody abolished FGF19-mediated activity in vitro and inhibited growth of colon tumor xenografts in vivo and effectively prevented hepatocellular carcinomas in FGF19 transgenic mice. The efficacy of the antibody in these models was linked to inhibition of FGF19-dependent activation of FGFR4, FRS2, ERK and beta-catenin. These findings suggest that the inactivation of FGF19 could be beneficial for the treatment of colon cancer, liver cancer and other malignancies involving interaction of FGF19 and FGFR4.

Study on steroids of Cacalia tangutica / 中国中药杂志

Seven steroids and two cumarins were isolated from the petroleum ether extract of the specie Cacalia tangutica of the family Compositae which were collected in Minhe county, Qinhai province of China. The structures were identified as Stigmast4-en-3beta, 6beta-diol (1), 24-ethyl-5alpha-cholestane-3beta, 5, 6beta-triol (2), 7beta- methoxy-stigmast-5-en-3beta-ol (3), Schleicherastatin 1 (4), Stigmast-5-en-3beta, 7alpha-diol (5), umbelliferone (6) and hydrangetin (7) by the means of chemical and modern spectroscopic analysis (MS, 1H-NMR, 13C-NMR, DEPT). The compounds 1-5 were isolated from Cacalia tangutica for the first time.

Genetic mapping in mice identifies DMBT1 as a candidate modifier of mammary tumors and breast cancer risk.

Low-penetrance breast cancer susceptibility alleles seem to play a significant role in breast cancer risk but are difficult to identify in human cohorts. A genetic screen of 176 N2 backcross progeny of two Trp53(+/-) strains, BALB/c and C57BL/6, which differ in their susceptibility to mammary tumors, identified a modifier of mammary tumor susceptibility in an approximately 25-Mb interval on mouse chromosome 7 (designated SuprMam1). Relative to heterozygotes, homozygosity for BALB/c alleles of SuprMam1 significantly decreased mammary tumor latency from 70.7 to 61.1 weeks and increased risk twofold (P = 0.002). Dmbt1 (deleted in malignant brain tumors 1) was identified as a candidate modifier gene within the SuprMam1 interval because it was differentially expressed in mammary tissues from BALB/c-Trp53(+/-) and C57BL/6-Trp53(+/-) mice. Dmbt1 mRNA and protein was reduced in mammary glands of the susceptible BALB/c mice. Immunohistochemical staining demonstrated that DMBT1 protein expression was also significantly reduced in normal breast tissue from women with breast cancer (staining score, 1.8; n = 46) compared with cancer-free controls (staining score, 3.9; n = 53; P < 0.0001). These experiments demonstrate the use of Trp53(+/-) mice as a sensitized background to screen for low-penetrance modifiers of cancer. The results identify a novel mammary tumor susceptibility locus in mice and support a role for DMBT1 in suppression of mammary tumors in both mice and women.

Structural and functional characterization of the transforming growth factor-beta -induced Smad3/c-Jun transcriptional cooperativity.

Smads are intracellular proteins that act as central effectors for transforming growth factor-beta (TGF-beta) and related proteins from the activated receptor into the nucleus, where they regulate ligand-induced gene expression. AP-1 binding sites have been functionally linked to the transcriptional activation of various genes in response to TGF-beta. Accordingly, we have previously shown that the heteromeric complex of Smad3 and Smad4 synergizes with c-Jun/c-Fos at the AP-1 binding site of the collagenase I promoter to induce transcriptional activation in response to TGF-beta. Using the collagenase I promoter as model system, we have now investigated the role of the c-Jun and Smad3 interactions with the promoter DNA and have further characterized the physical basis of the c-Jun/Smad3 interaction in the transcriptional response. Mutational analyses of the c-Jun protein and the AP-1 binding site in the promoter revealed that the interaction of c-Jun with DNA is necessary for transcriptional activation by TGF-beta and Smad3. Similar analyses of Smad3 and the Smad binding sites revealed that binding of Smad3 to DNA is also required, but that its DNA sequence-specific recognition is not essential. We also found that the basic leucine zipper domain of c-Jun and a short sequence close to the N terminus of Smad3 mediate their physical interaction, and that these regions are critical for their DNA-binding function. Our studies provide a basis for understanding the functional cooperativity of Smads with the diversity of transcription factors, which underlies the Smad-induced transcriptional activation in response to TGF-beta and related factors.

Multiple dose pharmacokinetics of artemether in Chinese patients with uncomplicated falciparum malaria.

Multiple dose pharmacokinetics of artemether and dihydroartemisinin were investigated in chinese patients treated for malaria. They received over 2 days either 4 x 80 mg artemether orally (n = 48) or 4 x 80-480 mg co-artemether (n = 40), a combination of artemether and lumefantrine (benflumetol). Lag time = 0.48 h (mean), Cmax after first dose = 157 ng/ml, t(max) = 1.73 h and elimination half-life = 1.16 h. The lag and absorption times were 0.5 h longer for co-artemether compared with artemether. Dihydroartemisinin paralleled artemether pharmacokinetics. Artemether Cmax after the last dose was one-third of the Cmax after the first dose while, inversely, dihydroartemisinin Cmax increased over time. We suggest that auto-induction of gut mucosa enzymes and/or liver enzymes causes a time-dependent increase in first-pass metabolisation of artemether.

Cloning and characterization of a novel gene encoding a putative transmembrane protein with altered expression in some human transformed and tumor-derived cell lines.

Identification and characterization of genes expressed in normal cells and decreased in their malignant counterparts is an important method for detecting candidate tumor suppressors. Using differential display of mRNAs from nontumorigenic infinite life span human fibroblast cell strain MSU-1.1 and an isogenic fibrosarcoma-derived cell line, 6A/SB1, which was derived from chemical carcinogen transformed MSU-1.1 cells, we identified a novel gene, ST7, showing sixfold lower expression in 6A/SB1 cells compared with parental MSU-1.1 cells. Molecular cloning of a near full-length cDNA revealed that the novel gene encodes a putative transmembrane protein composed of 859 amino acids: the 492 N-terminal amino acids including a fivefold cysteine-rich repeat of 40 amino acids homologous to the ligand binding repeat of the known low density lipoprotein receptor, a 24 hydrophobic amino acid stretch spanning the plasma membrane, and a C-terminal domain of 343 residues. ST7 is located on human chromosome 8, band q22.2-23.1, the same locus as the genes involved in acute myeloid leukemia and a locus of high polymorphism in cancer biopsies. The ST7 gene is widely expressed in normal human tissues and is particularly abundant in human heart and skeletal muscle. Northern analysis of 15 tumor cell lines derived from patients and 16 cell lines established from tumors formed in athymic mice by MSU-1.1 cells transformed in culture by various methods showed that 16 of the 31 cell lines have low or undetectable levels of ST7 mRNA. Furthermore, Western blotting analysis using a specific anti-peptide antibody demonstrated that the level of ST7 protein is high in normal fibroblasts and low in 12 sarcoma-derived cell lines tested. Altered expression of ST7 appears to occur at both the transcriptional and post-transcriptional levels. These studies are a first step in characterizing a novel putative receptor protein, whose expression is downregulated in some malignantly transformed cells, and which may play an important role in the transformation process of these cells.

Expression of rainbow trout growth hormone cDNA in Saccharomyces cerevisiae.

Rainbow trout growth hormone cDNA was modified by polymerase chain reaction (PCR). The modified cDNA was subcloned into the E. coli-yeast shuttle vector pMA91 under the yeast PGK promoter, and transformed into Saccharomyces cerevisiae Y33 to construct an expression strain Y33 (pMArGH16). The recombinant gene could express the growth hormone peptide (about 3% of the total yeast proteins) in Y33 (pMArGH16). The expression product was used as a supplement to feed Tilapia fingerlings. The result showed that the recombinant fish GH could significantly enhance the growth of Tilapia fingerlings.

Suppression of anchorage-independent growth and matrigel invasion and delayed tumor formation by elevated expression of fibulin-1D in human fibrosarcoma-derived cell lines.

Using differential display, we identified an mRNA that is markedly down-regulated in cell line 6A/SB1, derived from a fibrosarcoma formed in an athymic mouse following injection of carcinogen-transformed MSU-1.1 cells. The nontumorigenic parental cell strain, MSU-1.1, expresses high levels of this mRNA. Sequencing of the corresponding cDNA fragment revealed that it corresponded to an expressed sequence tag, which ultimately led to its identification as the fibulin-1D gene. Fibulin-1 is a cysteine-rich, calcium-binding extracellular matrix and plasma protein, which has four isoforms, A-D, derived from alternative splicing. Northern and Western blotting analysis of 16 cell lines established from tumors formed in athymic mice by MSU-1.1-derived cell strains independently transformed in culture showed that 44% exhibited low level or lack of expression of fibulin-1D mRNA and protein. In a similar analysis of 15 malignant cell lines derived from patients, 80% showed low level or no expression. To study the role of fibulin-1D in transformation, we transfected 6A/SB1 cells and a human fibrosarcoma-derived cell line (SHAC) with a fibulin-1D cDNA expression construct. Transfectants displaying high levels of fibulin-1D were isolated and characterized. Elevated expression of fibulin-1D led to reduced ability to form colonies in soft agar and reduced invasive potential as tested in a matrigel in vitro invasion assay. Furthermore, expression of fibulin-1D resulted in a markedly extended latency in tumor formation in athymic mice. These results indicate that low expression of fibulin-1D plays a role in tumor formation and invasion.

Successful replantation in 10-digit complete amputations.

We report a case of successful replantation of 10-digit complete amputations. The conditions of the wounds over the ends of the 10 amputated digits are bound to vary; some are too severe to be replanted, while others may be light and easy for replantation. We reported a case of 10-digit complete amputations, with severe avulsion injuries over four fingers. Appropriate measures were taken during surgery, and the replanted digits all survived. The replantation operation was done by a team of surgeons, although the surgeons were physically challenged by the long and difficult procedures. However, it was the great pleasure of the surgeons to work hard and whole-heartedly for the benefit of the patient. Postoperative physiotherapy and exercises were done. After 6 months, the functions of both hands had been largely recovered. Complete amputations of 10 digits as a rule result in multiple complicated wounds. The replantation procedure for these conditions is not an easy endeavor because there are numerous blood vessels that need to be anastomosed, the time of the operation will be dreadfully long, and also there is the danger of the occurrence of vascular crises during the postoperative period. It is difficult to achieve successful survival of all 10 amputated digits after replantation.