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A k-tree is a tree with maximum degree at most k. In this paper, we give a sharp degree sum condition for a graph to have a spanning k-tree in which specified vertices have degree less than t, where 1 ≤ t ≤ k. We denote by σk(G) the minimum value of the degree sum of k independent vertices in a graph G. Let k ≥ 2, s ≥ 0 and 1 ≤ t ≤ k be integers, and suppose G is an (s + 1)-connected graph with σk(G) ≥∣G∣ + (k − t)s − 1. Then for any s specified vertices, G contains a spanning k-tree in which every specified vertex has degree at most t. This improves a result obtained by Matsuda and Matsumura.

The patient was a 62-year-old man diagnosed as having prostatic extra-gastrointestinal stromal tumor (EGIST) who was treated with imatinib. No recurrence or metastasis was found after a 6-month follow-up. We identified 14 cases of prostatic primary EGIST in PubMed and summarized these cases with our case. The patients’ ages ranged from 31 to 78 years (average: 53.6 years), and most patients’ prostate-specific antigen (PSA) concentrations were within normal limits (92.9%, 13/14). All patients underwent imaging examinations; prostatic masses measured 6 to 14.2 cm (mean: 9.43 cm), and imaging excluded secondary prostatic masses from the intestinal tract. By immunohistochemical staining, the tumors were positive for cluster of differentiation (CD)117 (71.4%, 10/14), DOG1 (100%, 7/7), and CD34 (100%, 14/14), and negative for smooth muscle actin (SMA) (71.4%, 10/14), desmin (100%, 11/11), and S100 (100%, 12/12). Treatment depended on the results of the gene mutation detection as well as the risk estimation according to tumor size and microscopic mitotic rates (>5 per 50 high-power fields: 60%, 6/10). Among the 12 patients with reported outcomes, nine achieved good results (no recurrence or metastasis), one achieved reduced mass volume, one experienced recurrence, and one died.

Nitrogen and phosphorus are the leading causes of water eutrophication, and it is challenging to remove nitrogen and phosphorus effectively through a single water remediation method. In this study, an aerobic denitrifying bacterium (AD-19) isolated from eutrophic water was used to construct an immobilized biofilm and combined with Phoslock

Imaging-guided percutaneous microwave ablation (MWA) with high thermal efficiency comprises rapid, successful management of small renal cell carcinomas (RCCs) in selected patients. Ultrasound Committee of Chinese Medical Association, Interventional Oncology Committee of Chinese Research Hospital Association developed evidence-based guidelines for MWA of RCCs after systematically reviewing the 1969–2019 literature. Systematic reviews, meta-analyses, randomized controlled trials, cohort, and case–control studies reporting MWA of RCCs were included and levels of evidence assessed. Altogether, 146 articles were identified, of which 35 reported percutaneous MWA for T1a RCCs and 5 articles for T1b RCCs. Guidelines were established based on indications, techniques, safety, and effectiveness of MWA for RCCs, with the goal of standardizing imaging-guided percutaneous MWA treatment of RCCs.Key pointsMicrowave ablation is recommended for managing small renal cell carcinoma in selected patients.Imaging protocols are tailored based on the procedural plan, guidance, and evaluation.Patient’s selection evaluation, updated technique information, clinical efficacy, and complications are recommended to standardize management.A joint task force (multidisciplinary team) summarized the key elements of the standardized report. Microwave ablation is recommended for managing small renal cell carcinoma in selected patients. Imaging protocols are tailored based on the procedural plan, guidance, and evaluation. Patient’s selection evaluation, updated technique information, clinical efficacy, and complications are recommended to standardize management. A joint task force (multidisciplinary team) summarized the key elements of the standardized report.

In the paper, we propose a surface wave suppression method in time-frequency domain based on the wavelet transform, considering the characteristic difference of polarization attributes, amplitude energy and apparent velocity between the effective signals and strong surface waves. First, we use the proposed method to obtain time–frequency spectra of seismic signals by using the wavelet transform and calculate the instantaneous polarizability at each point based on instantaneous polarization analysis. Then, we separate the surface wave area from the signal area based on the surface-wave apparent velocity and the average energy of the signal. Finally, we combine the polarizability, energy, and frequency characteristic to identify and suppress the signal noise. Model and field data are used to test the proposed filtering method.

To suppress the strong noise in seismic data with wide range of amplitudes, commonly used methods often yield unsatisfactory denoising results owing to inappropriate thresholds and require parametric testing as well as iterations to achieve the anticipated results. To overcome these problems, a data-driven strong amplitude suppression method based on the decibel criterion in the wavelet domain (ISANA) is proposed. The method determines the denoising threshold based on the decibel criterion and statistically analyzes the amplitude index rather than the abnormally high amplitudes. The method distinguishes the frequency band distributions of the valid signals in the time–frequency domain based on the wavelet transformation and then calculates thresholds in selected time windows, eventually achieving frequency-divided noise attenuation for better denoising. Simulations based on theoretical and real-world data verify the adaptability and low dependence of the method on the size of the time window. The method suppresses noise without energy loss in the signals.

For non-negative integers i, j and k, we denote the generalized net as N i,j,k , which is a triangle with disjoint paths of length i, j and k, attached to distinct vertices of the triangle. In this paper, we prove that every 3-connected {K 1,3,N 8−i,i,1}-free graph is hamiltonian, where 1 ≤ i ≤ 4.

The depolymerization behaviors of Xianfeng lignite (XL) in ionic liquid 1-butyl-3-methyl-imidazolium trifluoromethanesulfonate ([Bmim]OTf) was studied in this paper. Also, the thermal extracts and residues obtained from thermal depolymerization of XL at different extraction conditions were analyzed. It was found that [Bmim]OTf had good performance in the depolymerization of XL when the extraction was carried out at 280°C for 4 h with [Bmim]OTf/XL at 3.5. The extraction yield of XL with [Bmim]OTf defined as tetrahydrofuran solubles (THFS) reached to 20.1%. The depolymerization of XL with [Bmim]OTf was promoted markedly with the addition of tetralin (THN), H 2 and ZnCl 2 catalyst, and the yield of THFS reached to 30.0%, 36.9% and 46.8%, respectively. The results showed that the amount of hydrogen bonds in thermal extracts increased significantly with the addition of THN. The contents of aromatics and aliphatic hydrocarbon were enhanced greatly under the existence of H 2 and ZnCl 2 .

Background: This study was undertaken to investigate the expression of guanyl nucleotide-releasing protein for Ras 3 (RasGRP3) in the cell lines and tissues in BPH and prostate cancer (PCa), as well as its associations with cancer invasion and prognosis in prostate carcinomas. Methods: Expression analysis of RasGRP3 was accomplished using immunohistochemical staining of PCa and BPH tissues. Pearson's χ2 test was used to analyze the association between RasGRP3 expression and specific clinical parameters. Survival and PSA relapse curves were evaluated using the Kaplan–Meier curves and log-rank tests, and the differences were assessed using the Cox regression methods. In addition, human PCa cell lines PC-3, DU145, LNCaP, PC3M-1E8, PC3M-2B4 and BPH-1 were examined for expression of RasGRP3 using western blot and quantitative polymerase chain reaction (Q-PCR) analysis. After PC-3 cells were transfected by small interfering RNA targeting RasGRP3, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and migratory assays were employed to determine the vitality and aggressive capability of tumor cell in vitro. Results: Expression of RasGRP3 was significantly correlated (P=0.038 and P=0.021) with Gleason score (⩽6 versus ⩾7) and T stage (T1–T2 versus T3–T4), respectively. PCa with RasGRP3-positive expression may increase the risk of PSA recurrence and decrease cancer-specific survival (P=0.0291 and P=0.0044). The expression of RasGRP3 was also associated with PSA recurrence and cancer-specific survival in univariate (P

N-methyl pyrrolidonium hydrosulfate ([Hnmp]HSO4) was synthesized and characterized. N-butyl acetate was synthesized from acetic acid and n-butanol with [Hnmp]HSO4 as catalyst in a catalytic rectification column 30 mm in diameter and 2.5 m in height. Effects of catalyst dosage, bottom temperature, feeding ratio of acetic acid to n-butanol, feeding method and reflux ratio on esterification and rectification were investigated. Under optimal condition which taking both energy cost and product quality into account: catalyst dosage was 0.5% of feed weight, bottom temperature was 120 °C, mole ratio of acetic acid to butanol was 1:1.2, acetic acid and butanol were mixed and fed above second section of packing, feeding rate was 150 mL/h and reflux ratio was 4, mass fraction of n-butyl acetate in bottom could reach 92.49 %.

X. Deng et al. proved Chvatal’s conjecture on maximal stable sets and maximal cliques in graphs. G. Ding made a conjecture to generalize Chvatal’s conjecture. The purpose of this paper is to prove this conjecture in planar graphs and the complement of planar graphs.

Microcystins, one type of the cyanobacterial toxins, show a broad range of hazardous effects on other organisms. Most of the researches on the toxic effects of microcystins have involved in animals and higher plants. Little work, however, has been done on evaluating the mechanisms of microcystin toxicity on algae. In this study, the toxicological effects of microcystin-RR (MC-RR) on the cyanobacterium Synechococcus elongatus were investigated. For this purpose, six physio-biochemical parameters (cell optical density, reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GSH-Px) and glutathione S-transferase (GST)) were tested in algal cells when exposed to 100 μg−1 microcystin-RR. The results showed that the growth of Synechococcus elongatus (expressed as optical density) was significantly inhibited compared with the control. At the same time, the treated algae exhibited a pronounced increase in production of ROS and MDA after 6 days exposure to microcystin-RR. Significant changes in GSH levels and GSH-Px, GSH activities were also detected in algal cells, with higher values being observed in the toxin treated algae after 6 days exposure. GST activities in the treated algae exhibited a decline after exposure and rapid augmentation on day 3, thereafter, they kept at a high level when compared to the control group. GSH contents and GSH-Px activities were also significantly raised in the toxin-treated algae cells from day 3, but they showed a sharp decrease on day 4, which was the onward of cell proliferation. These results suggested that oxidative stress manifested by elevated ROS levels and MDA contents might be responsible for the toxicity of microcystin to Synechococcus elongatus and the algal cells could improve their antioxidant ability through the enhancement of enzymatic and non-enzymatic preventive substances.

A graph g is k-ordered Hamiltonian, 2 ≤ k ≤ n, if for every ordered sequence S of k distinct vertices of G, there exists a Hamiltonian cycle that encounters S in the given order. In this article, we prove that if G is a graph on n vertices with degree sum of nonadjacent vertices at least $$n+{{3k - 9} \over 2}$$ , then G is k-ordered Hamiltonian for $$ k = 3,4, \cdots ,{\left\lfloor {\frac{n} {{19}}} \right\rfloor } $$ . We also show that the degree sum bound can be reduced to $$ n + 2{\left\lfloor {\frac{k} {2}} \right\rfloor } - 2 $$ if $$ \kappa {\left( G \right)} \geqslant \frac{{3k - 1}} {2} $$ or δ(G) ≥ 5k − 4. Several known results are generalized.

To explore the relationship between occupational factors and the incidence of bladder cancer.The present research was based on a hospital-based case-control study. The cases were frequency matched. The non-conditional Logistic regression was used to calculate the odds ratio (OR) of each occupation.The OR of business and administration professionals, male electricians and electronic workers were 3.88 and 7.40; the OR of janitors and helpers was 0.21; the OR of handcrafted and printing clerks was 0.71, but there was no significant difference.Business and administration professionals, male electricians and electronic workers tend to have bladder cancer. The occupation of janitors and helpers has a protective effect on bladder cancer while the occupation of printing clerks shows no statistical significance on bladder cancer.

Bladder urothelial cancer has been diagnosed at an increasing rate among young adults in China while the clinical outcomes remain highly controversial. To optimize the management of young patients with bladder cancer, we examined whether bladder urothelial cancer in young patients behaved differently from that in the elder patients.From 1994 to 2008, a database of bladder urothelial cancer patients at a major tertiary medical center was retrospectively reviewed. The clinical and pathological parameters of patients who were less than 40 years of age and a series of patients older than 40 years of age as the control group during the same period were compared. A survival analysis was performed using the Kaplan-Meier method and log-rank test, and Cox regression was performed to identify clinical parameters that affected the clinic outcomes.Young bladder cancer patients had a lower male-to-female ratio and were less likely to have advanced stages and high-grade cancers at the initial diagnosis. Tumors in young bladder cancer patients tended to be less multifocal at diagnosis. In addition, young patients had a lower recurrence rate and longer recurrence interval than older patients. The Kaplan-Meier curve and Log-rank test showed that young patients had significantly better cancer specific survival than old patients. The univariate and multivariate Cox regression analysis revealed that tumor grade is the sole predictor for tumor recurrence in young patients.Young patients with bladder cancer have favorable pathological features and clinical outcomes than older patients. These findings argue for more conservative management approaches for young patients with bladder cancer.

To explore the clinical, pathological features and prognosis of patients with chromophobe renal cell carcinoma.From January 1998 to January 2008, clinical data of 29 patients with chromophobe renal cell carcinoma including clinical manifestations, imaging examinations, treatment models, pTNM stages and follow-up results, were summarized to investigate its features and prognosis.All cases had no obvious clinical and preoperative imaging presentation. There were 23 patients underwent radical nephrectomy, and 6 cases underwent nephron sparing surgery. Postoperative pathological findings confirmed the diagnosis of chromophobe renal cell carcinoma. Macroscopically, the cut surface of the tumors were generally beige in color. Histologically, it showed polygonal chromophobe cells and small round eosinophilic cells with eccentric hyaline degeneration. These tumor cells had a clear and sharp membrane, lightly stained abundant cytoplasm with a fine reticular translucent pattern and irregular nuclei. And a perinuclear halo was often seen in these cells. Histochemically, the tumor cells generally show a diffuse and strong reaction for CK-8 with a negative expression of Vimentin. The pTNM stages of the tumor were as follows, pT1N0M0 in 11 cases, pT2N0M0 in 8 cases, pT3aN0M0 in 5 cases, pT1N1M0 in 3 cases, pT2N1M0 in 2 cases. Twenty-six cases of patients were followed up (24 to 144 months, with an average of 90 months), 3 cases died of cardio-cerebrovascular disease, and local recurrence involved in 6 cases with reoperation in 4 cases, as well as distant metastasis in 1 case. Twenty-one cases survived with tumor-free. The statistical results indicated that the survival rates of the patients with chromophobe renal cell carcinoma in five years and ten years were 83.9%, 77.9%, respectively, compared with renal cell carcinoma of the same stage 63.8% and 49.9% at the same periods, and there is no difference in the survival rate of five years (P0.05) but significant difference in that of ten years (P0.01).Chromophobe renal cell carcinoma is a morphologically uncommon subtype of renal cell carcinoma with the good prognosis. Definite diagnosis depends on its typical pathological feature. Radical nephrectomy is the first choice for the treatment of chromophobe renal cell carcinoma.

One aspect of chemotherapy insensitivity and resistance results from induction of epidermal growth factor receptor (EGFR) internalization and initial DNA damage repair in response to DNA-damaging stimuli, such as cisplatin (CDDP). Previously, we found that leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1), as one of the natural ligands of EGFR, could combine with and down-regulate the expression of EGFR in bladder cancer cells. This finding interested us and we hypothesized that LRIG1 could be a novel candidate for facilitating cisplatin-induced bladder cancer cell lesions. To investigate this further, we overexpressed LRIG1 with an adenovirus vector in EJ/T24 bladder cancer cells and investigated total EGFR, nuclear expression of phosphorylated EGFR (pEGFR) and cell lesions with exposure to CDDP. CDDP-induced nuclear pEGFR levels accumulated with time and were decreased by LRIG1 overexpression. LRIG1-transduced cells treated with CDDP had more severe DNA damage, cellular apoptosis, growth inhibition and reversal of invasion. These preclinical studies indicate that LRIG1 may represent a new therapeutic approach to improve the response of bladder cancer to chemotherapy through a novel pathway.

Aims A study was conducted to compare the efficacy and complications of tension-free vaginal tape (TVT), transobturator vaginal tape inside-out TVT-0 and transobturator vaginal tape out-inside (TOT) procedures for the surgical treatment of female stress urinary incontinence SUI. Methods This study is a prospective randomized trial involving 187 women with primary SUI; 77 received TVT, 65 received TVT-O, and 45 patients were treated with TVT-0 between June 2002 and December 2009. Before the operation, a complete medical history was taken and a gynecologic examination was performed, including subjective symptoms, history and urodynamic studies. Postoperative data included mean operation time, days of hospitalization, postoperative complications and efficacy evaluation. Therapeutic effect was assessed by presence or absence of incontinence when abdominal pressure increased. Results The history, physical examination and urodynamic studies among the three groups have no significant difference (P>0.05). The total cure rate was 91.4%. The cure rate in TVT, TVT-O and TOT groups were 90.9%, 92.3% and 91.1%, respectively. There was no significant difference (P>0.05). Mean operative time showed no significant difference between TVT-O and TOT groups, but both were significantly shorter than TVT group (26.90±16.80, 20.00±13.50 vs 48.20±21.90). The mean postoperative hospital stay showed no significant difference between TVT and TVT-O groups, but both groups were significantly longer than TOT group. Mean postoperative hospital stay of TVT, TVT-O and TOT were 5.00±2.40 days, 4.00±2.20 days and 2.30±0.80 days, respectively. The complication rate in TVT, TVT-O and TOT groups was 15.60%, 9.20% and 8.90%, respectively. In TVT group, 4 patients experienced bladder perforation, postoperative dysuria or retention occurred in 7 cases and was cured by urethral dilation, hematomas of retropubic space in 1 patient. No bladder injury occurred in TVT-O and TOT group, 3 patients had postoperative dysuria or retention and 3 patients had transient dysfunction of both lower limbs postoperatively in TVT-O group, 2 patients had postoperative dysuria or retention and 2 patients had transient dysfunction of both lower limbs postoperatively in TOT group. Summary The three tension-free urethral suspension techniques have similar efficacy, all of them are safe and effective procedures for the treatment of female SUI. Compared with TVT, TVT-O and TOT are simpler, less invasive and have fewer complications.

To investigate the effect of B7-H1 blockade on proliferation, activation, and antitumor immunity of CD3AK cells.CD3AK cells were induced by stimulation of normal human peripheral blood lymphocytes with CD3 mAbs. Then the cells were cultured with anti-B7-H1 mAbs to block B7-H1 pathway. The proliferation efficiency of CD3AK cells was measured by 3H-thymidine incorporation assay and the concentrations of IFN-gamma, TNF-alpha and IL-10 were measured by ELISA method. Meanwhile the killing activity of CD3AK cells on bladder cancer cell line BIU-87 was measured by MTT method.Blockade of B7-H1 greatly promoted the proliferation of CD3AK cells and extended the survival time of CD3AK cells in vitro. It also enhanced IFN-gamma, TNF-alpha secretion but suppressed IL-10 secretion. And the cytotoxic effect of CD3AK cells on BIU-87 cells were significantly enhanced.Blockade of B7-H1 can promote and retain the proliferation and activation of CD3AK cells. It can also improve the antitumor immunity mediated by CD3AK cells. The manipulation of B7-H1 may become a beneficial target for immunotherapy in tumors.

To evaluate the efficacy and safety of gonadotropin-releasing hormone analogue (GnRHa) triptorelin 11.25 mg 3-month sustained release formulations in the treatment of metastatic prostate cancer.From January 2004 to March 2006, a randomized, parallel-controlled, multicenter clinical trial was conducted. One hundred and twenty-seven patients with documented metastatic prostate cancer were randomized to receive one injection of the 11.25 mg formulation triptorelin (n = 65) or three injections at 28-day intervals of the 3.75 mg formulation (n = 62). Changes from baseline of TPSA, prostate volume, testosterone, LH, FSH, PRL and estradiol were assessed over 3 months. Changes of the metastatic lesions were also observed and evaluated. The occurrences of adverse events were evaluated as well.After 3 months treatment, total PSA level decreased significantly from baseline both in 11.25 mg group and 3.75 mg group. At 30, 60 and 90 days, TPSA (median level) declined from 164.55 microg/L into 11.34, 4.12, 3.89 microg/L in 11.25 mg group, and from 101.38 microg/L into 6.88, 2.41, 2.57 microg/L in control group respectively. The patients ratio with over 90% decreasing from TPSA baseline were 78.6% and 75.5% respectively in two groups (P = 0.700). Prostate volume were also decreased significantly in both groups, median volume declined from 48.0 mm(3) into 21.5 mm(3) in 11.25 mg group and from 45.0 mm(3) into 21.0 mm(3) in 3.75 mg group. No significant differences were found between the two groups in changes of TPSA (P = 0.601) and prostate volume (P0.05). Both formulations were able to induce castration levels, 0.31 microg/L in 11.25 mg group and 0.26 microg/L in 3.75 mg group (P0.05). 13.8% and 17.7% of adverse events were recorded respectively in two groups, and no differences were found (P = 0.547).As a new long-acting sustained release formulation, triptorelin 11.25 mg is comparable to triptorelin 3.75 mg formulation in the aspect of efficacy and safety for the treatments of metastatic prostate cancer.

To study the inhibitory effect of matrine on the proliferation of the prostate cancer cell line LNCaP and the expression of the androgen receptor (AR).LNCaP cells were treated with matrine at the concentration of 0.5, 1.0, 1.5, 2.0 and 3.0 g/L for 12, 24 and 36 hours, the cell growth activity determined by MTT colorimetry and trypan blue staining at 36 hours, the cell cycle changes detected by flow cytometry and the expression of AR by Western blot at 24 hours.Matrine suppressed the in vitro growth of the androgen-sensitive prostate cancer cell line LNCaP in a time- and dose-dependent manner, blocked the cell cycles in the G2/M phase and decreased the expression of AR in the cell line in a dose-dependent manner (P0.01).Matrine can significantly inhibit the in vitro growth of NCaP cells by down-regulating the expression of AR and blocking cell cycles.

To study the effects of curcumin on the expression of the vascular endothelial growth factor (VEGF) and androgen-independent prostate cancer cell line PC-3, and to explore its anticarcinogenic mechanism.PC-3 cells were treated with curcumin at the concentration of 0, 6.25, 12.5, 25 and 50 micromol/L respectively. Then the cell activity was assayed by dyed rate of Typan blue and MTT at 12, 24, 36, 48, 72 and 96 hours, the cell cycle and morphological changes observed by flow cytometry (FCM) and electronic microscopy at 24 hours, the VEGF mRNA expression measured by semi-quantitative RT-PCR, and the secreting protein levels of VEGF in the supernatants determined by enzyme-linked immunosorbent assay (ELISA).The growth of PC-3 cells was suppressed obviously by curcumin in a dose- and time-dependent manner in vitro. There were significant differences in inhibition rate among different concentration and time groups (P0.01). Furthermore, curcumin arrested the cell cycle of PC-3 cells in the G2/M phase in a dose-dependent manner (P0.01). The percentages of apoptotic cells were significantly higher in different concentration groups than in the controls (P0.01). Apoptosis-associated morphological changes were observed in PC-3 cells at 24 hours, and a marked decline in the expression of VEGF was noted after the exposure to different concentrations of curcumin within 24 hours.Curcumin can suppress the growth of PC-3 cells, promote their apoptosis and arrest their cell cycle in the G2/M phase, and reduce the expression of VEGF mRNA and proteins, which may sever to explain its inhibitory effect on tumor and angiogenesis.

To evaluate the effects of antisense oligodeoxynucleotide (ASODN) of survivin gene on taxol-induced apoptosis in bladder cancer cells.A survivin ASODN eukaryotic vector pcDNA3-SVVas was transfected into human bladder cancer cells of the line BIU87 mediated by liposomal reagent (BIU87 SVVas cells). The mRNA expression of survivin was measured by RT-PCR. Blank plasmid pcDNA3 was transfected as control (BIU87 neo cells). The cell growth curve was drawn using trypan blue dye exclusion assay. MTT assay was used to investigate the sensibility of transfected cells to taxol. The BIU87 SVVas cells, BIU87 neo cells, and BIU87 cells were cultured in the culture fluid with taxol and then DNA gel electrophoresis, Hoechst nuclear staining and fluorescent microscopy, and annexin-propidium iodide (PI) staining were used to examine the cell apoptosis.Compared to BIU87 and BIU87 neo cells, the mRNA expression of survivin in the BIU87-SVVas cells was obviously reduced. The growth of the BIU87-SVVas cells was significantly reduced in comparison with the BIU87 (P0.05). The sensibility of BIU87 SVVas cells to taxol increased significantly shown by cell proliferation and MTT assays. Agarose gel electrophoresis of genomic DNA showed typical DNA ladder only in the BIU87 SVVas cells, but not in other cells. Hoechst nuclear staining and fluorescent microscopy showed that the nuclei of the BIU87 SVVas cells become condense. Annexin-PI test showed that the cell apoptosis rate of the BIU87-SVVas cells treated with taxol was significantly higher than those of the other groups.survivin antisense RNA enhances the taxol-induced apoptosis in the bladder cancer cells. This may lay an experimental foundation for further research of biotherapy in bladder cancer.

To prepare the tumor antigen peptide complex (HSP70-1d) of HSP70 and idiotype (Id) from SmIg ScFv fragment in patients with Chronic B cell leukemia (B-CLL), and to study the anti-tumor effect of cytotoxic T lymphocyte (CTL) induced by HSP70-Id complex-modified dendritic cell (DC) in vitro and explore their immune mechanism.Purified HSP70 was combined into peptide complex (HSP70-Id) with the prepared Id-ScFv from B-CLL cells in vitro by using biochemical technique. The plastic-adherent monocytes from human peripheral blood were cultured and induced into DC with rhGM-CSF and rhIL-4 using cell culture and separation technique. The cultured DC were harvested and pulsed by HSP70-Id complex. DC morphology was observed under converted phase microscope and its phenotype was characterized by FCM on 8th day as well as their secreting cytokines were measured. Host lymphocytes were stimulated by DC loaded with HSP70-Id complex and co-cultured in the medium containing IL-2. The activation and proliferation of lymphocytes were examined by MTr test, which was also used to assay cytotoxicity of CTL elicited by modified DC to Daudi, K562 and HepG2 tumor cells, and FCM analyzed the changes of T lymphocyte subsets.Mature DCs were obtained successfully, showing typical morphology and phenotypic properties, the expression ratio of cellular surface molecules, CD1a was 20% - 30%, CD83 was more than 72% , both CD86 and HLA-DR over-expressed obviously in the complex-loaded DC group secreting cytokines of Thl type, IL-12 and TNF-alpha. The culturing lymphocytes that were activated by modified DC could more effectively and specifically kill Daudi (71. 24%), but not K562 and HepG2 tumor cells. Results of FCM assay demonstrated that percentage of CD4+ and CD8+ T lymphocytes cocultured with complex-modified DC increased notably to 56.51% and 70.21%, respectively. CD4+ T/ CD8+ T proportion was changed from 1.49 to 0.81. The dose of peptide would be reduced to 1/50 if specific CTL induced by complex-modified DC instead of directly by peptide complex.DCs modified by HSP70-Id complex exhibit powerful biological activities, and could induce CTL to specific cytotoxicity against carcinoma cells. It might be produced by cooperation of CD4+ T, CD8+ T lymphocytes and DC. The results also suggested that DC modified by HSP70-Id complex can present antigen and induce CTL with high efficacy and specificity.

To evaluate the value of quantitative examination of MUC 1 in the urine of patients with bladder transitional cell carcinoma (BTCC).Urine samples were obtained from 31 patients with BTCC for quantification of MUC 1 content by immunoradiometric analysis. The urine samples were also examined in 10 patients with cystitis glandularis, 10 with benign urine disease and 10 healthy volunteers. The differences in urine MUC1 content were statistically measured between the groups, between cancer patients of different clinical stages and classes, between primary and recurrent cancer patients, and between measurements taken before and after operation.Urine MUC 1 was detected in all the patients. No significant differences were found between the groups, nor between patients with BTCC in all stages (P0.05), or between primary and recurrent cancer patients (P0.05). But MUC 1 contents showed significant difference before and after the operation in the cancer patients (P0.05).Urine MUC 1 can not serve as the marker to screen and diagnose BTCC, but it can be useful in therapeutic effect and prognostic evaluation. Specific oncogene markers are more significant than oncogene phenotype markers in clinical diagnosis and screen of BTCC.

Freshwater Microcystis may form dense blooms in eutrophic lakes. It is known to produce a family of related cyclic hepatopeptides (microcystins, MC) that constitute a threat to aquatic ecosystems. Most toxicological studies of microcystins have focused on aquatic animals and plants, with few examining the possible effects of microcystins on phytoplankton. In this study we chose the unicellular Synechococcus elongatus (one of the most studied and geographically most widely distributed cyanobacteria in the picoplankton) as the test material and investigated the biological parameters: growth, pigment (chlorophyll-a, phycocyanin), photosynthetic activity, nitrate reductase activity, and protein and carbohydrate content. The results revealed that microcystin-RR concentrations above 100 mug (.) L-1 significantly inhibited the growth of Synechococcus elongatus. In addition, a change in color of the toxin-treated algae (chlorosis) was observed in the experiments. Furthermore, MC-RR markedly inhibited the synthesis of the pigments chlorophyll-a and phycocyanin. A drastic reduction in photochemical efficiency of PSII (F-v/F-m) was found after a 96-h incubation. Changes in protein and carbohydrate concentrations and in nitrate reductase activity also were observed during the exposure period. This study aimed to evaluate the mechanisms of microcystin toxicity on a cyanobacterium, according to the physiological and biochemical responses of Synechococcus elongatus to different doses of microcystin-RR. The ecological role of microcystins as an allelopathic substance also is discussed in the article. (C) 2004 Wiley Periodicals, Inc.

Background The development of vectors for cell-specific gene delivery is a major goal of gene therapeutic strategies. Transferrin receptor (TfR) is an endocytic receptor and identified as tumor relative specific due to its overexpression on most tumor cells or tissues, and TfR binds and intakes of transferrin-iron complex. We have previously generated an anti-TfR single-chain variable fragments of immunoglobulin (scFv) which were cloned from hybridoma cell line producing antibody against TfR linked with a 20 aa-long linker sequence (G4S)4. In the present study, the anti-TfR single-chain antibody (TfRscFv) was fused to DNA-binding domain of the yeast transcription factor GAL4. The recombinant fusion protein, designated as TfRscFv-GAL4, is expected to mediate the entry of DNA-protein complex into targeted tumor cells. Results Fusion protein TfRscFv-GAL4 was expressed in an E. coli bacterial expression system and was recovered from inclusion bodies with subsequent purification by metal-chelate chromatography. The resulting proteins were predominantly monomeric and, upon refolding, became a soluble biologically active bifunctional protein. In biological assays, the antigen-binding activity of the re-natured protein, TfRscFv-GAL4, was confirmed by specific binding to different cancer cells and tumor tissues. The cell binding rates, as indicated by flow cytometry (FCM) analysis, ranged from 54.11% to 8.23% in seven different human carcinoma cell lines. It showed similar affinity and binding potency as those of parent full-length mouse anti-TfR antibody. The positive binding rates to tumor tissues by tissue microarrays (TMA) assays were 75.32% and 63.25%, but it showed weakly binding with hepatic tissue in 5 cases, and normal tissues such as heart, spleen, adrenal cortex blood vessel and stomach. In addition, the re-natured fusion protein TfRscFv-GAL4 was used in an ELISA with rabbit anti-GAL4 antibody. The GAL4-DNA functional assay through the GAL4 complementary conjugation with the GAL4rec-GFP-pGes plasmid to verify the GLA4 activity and GAL4rec-recognized specificity functions. It also shows the complex, TfRscFv-GAL4-GAL4rec-GFP-pGes, could be taken into endochylema to express the green fluorescent protein (GFP) with 8 to 10-fold transfection efficiency. Conclusions Results of our study demonstrated that the biofunctianality of genetically engineered fusion protein, TfRscFv-GAL4, was retained, as the fusion protein could both carry the plasmid of GAL4rec-pGes and bind TfR on tumour cells. This product was able to transfect target cells effectively in an immuno-specific manner, resulting in transient gene expression. This protein that can be applied as an effective therapeutic and diagnostic delivery to the tumor using endogenous membrane transport system with potential widespread utility.

A method is presented to get the dynamic characteristics of a multi-DOF rigid body structure from the measured vibration data. These characteristics include the mass, center of gravity, moment of inertia and the principal axes of the body, stiffness, damping, the center of stiffness in any direction, moment of stiffness, center of damping and moment of damping, and the principal axes of the supports. The method is capable of representing structures with various DOFs, from 2 up to 6. The mass M, stiffness K, and damping C (or M−1K and M−1C) matrices are obtained first based on measured vibration data by Data Dependent Systems (DDS) methodology which is accurate even for highly damped structures. Relationships between the physical characteristics and these matrices have been derived so that the physical characteristics can be obtained by solving simultaneous linear equations. It is also possible to determine the characteristics up to 5 individual springs and dashpots to provide a basis for improvement in the structure’s performance.