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Purpose: To investigate the effect of combinational delivery of urokinase-type plasminogen activator (uPA) and hepatocyte growth factor (HGF) genes on hepatic fibrosis., Methods: Replication-deficient adenoviral vectors expressing either human HGF (AdHGF) or uPA (AduPA) were generated. HGF gene was transferred into primary cultured hepatocytes and uPA gene to hepatic stellate cell (HSC) to investigate the effect on the biological character of cells. Combinational adenoviruses were infused into hepatic fibrosis rats. Serum markers as well as histological and immunohistochemical examination were carried out to test the reversal of hepatic fibrosis., Results: Transfection of exogenous HGF gene induced expression of c-met/HGF receptor and stimulated hepatocyte proliferation. uPA gene delivered into HSC decreased the amount of collagen types I and III accompanied with the increased expression of matrix metalloproteinase-2. In vivo, the area of extracellular matrix in the fibrotic liver decreased to 72% in AdHGF-treated rats (P<0.01), 64% in the AduPA-treated group (P<0.01), and 51% in bi-genes transfection (P<0.01), compared with that of the controls. Moreover, immunohistochemical staining of collagen types I and III revealed that combinational genes delivery exerted more effect on reversal of hepatic fibrosis than mono-gene transfection., Conclusions: Our study indicated that simultaneous delivery of two antifibrotic genes could confer synergistic effect on hepatic fibrosis.

An efficient biologically enhanced sampling geometric docking method is presented based on the FTDock algorithm to predict the protein-protein binding modes. The active site data from different sources, such as biochemical and biophysical experiments or theoretical analyses of sequence data, can be incorporated in the rotation-translation scan. When discretizing a protein onto a 3-dimensional (3D) grid, a zero value is given to grid points outside a sphere centered on the geometric center of specified residues. In this way, docking solutions are biased toward modes where the interface region is inside the sphere. We also adopt a multiconformational superposition scheme to represent backbone flexibility in the proteins. When these procedures were applied to the targets of CAPRI, a larger number of hits and smaller ligand root-mean-square deviations (RMSDs) were obtained at the conformational search stage in all cases, and especially Target 19. With Target 18, only 1 near-native structure was retained by the biologically enhanced sampling geometric docking method, but this number increased to 53 and the least ligand RMSD decreased from 8.1 A to 2.9 A after performing multiconformational superposition. These results were obtained after the CAPRI prediction deadlines.

Purpose: To investigate the safety, efficacy and feasibility of using high-intensity focused ultrasound (HIFU) as a non-invasive treatment for patients with breast cancer., Patients and Methods: Twenty-two patients with breast cancer were enrolled into this non-randomized prospective trial. Disease TNM stage was classified as stage I in 4 patients, stage II(A) in 9 patients, stage II(B) in 8 patients, and stage IV in 1 patient. Tumor size ranged from 2 to 4.8 cm in diameter (mean 3.4 cm). All patients received chemotherapy, radiation and tamoxifen, following HIFU for the primary lesions. Outcome measures included radiological and pathologic assessment of the treated tumor, cosmesis, and local recurrence. A cumulative survival rate is calculated by using the Kaplan-Meier method., Results: No severe complications were encountered after HIFU. Post-operative imaging demonstrated positive response and regression of all treated lesions. Follow-up biopsy revealed coagulation necrosis of target tumor and subsequent replacement by fibroblastic tissue. After a median follow-up of 54.8 months, 1 patient died, 1 was lost to follow-up, and 20 were still alive. Two of 22 patients developed local recurrence. Five-year disease-free survival and recurrence-free survival were 95% and 89%, respectively. Cosmetic result was judged as good to excellent in 94% of patients., Conclusions: HIFU treatment is safe, effective, and feasible for patients with breast cancer. But, large-scale, multiple-center clinical trials will be needed to determine the future role of this novel modality.

Objective: To investigate the effects of low-intensity pulsed ultrasound in repairing injured articular cartilage., Methods: Ten adult New Zealand rabbits with bilateral full-thickness osteochondral defects on the cartilage surface of intercondylar fossas were used in this study. The wounds in the left knees were treated with low-intensity pulsed ultrasound as the experimental group. The right knees received no treatment as the control group. All the animals were killed at 8 weeks after injury and the tissues in the wounds were collected for gross appearance grading, histological grading and proteoglycan quantity., Results: The scores of the gross appearance grades, histological grades and the optical density of toluidine blue of the tissues in the experimental group were significantly higher than those of the controls at 8 weeks after injury (P < 0.05)., Conclusions: Low-intensity pulsed ultrasound can accelerate the repair of injured articular cartilage.

The (bis)dansylated sulfonamide 1,2-C6H4(NHSO2C10H6-5-N(CH3)2)2 (1) extracted Pb(II) selectively from water into 1,2-dichloroethane via an ion-exchange mechanism and showed fluorescence quenching upon Pb(II) extraction. The distribution ratios for metal extraction (determined by ICP-MS) for Pb(II) were 133-1410 times higher than those for other metal cations [Co(II), Ni(II), Cu(II), Zn(II), and Cd(II)] under identical conditions. Fluorescence quenching was observed upon Pb(II) extraction, which was dependent on Pb(II) concentration. The monodansylated control, C6H5NHSO2C10H6-5-N(CH3)2 (2), showed neither extraction nor quenching, indicating that the fluorescence effects are a direct result of Pb coordination to 1. The observed selectivity for Pb(II) is ascribed to the formation of a low-coordinate binary Pb(II)-Sulfonamido complex in the organic phase.

Purpose: To evaluate ultrasonographically (US)-guided high-intensity focused ultrasound ablation combined with transcatheter arterial chemoembolization (TACE) in the treatment of stage IVA hepatocellular carcinoma (HCC)., Materials and Methods: Institutional review board approval and informed consent were obtained. From November 1998 to May 2000, 50 consecutive patients with stage IVA HCC (TNM classification, T4N0-1M0) were alternately enrolled in one of two treatment groups: group 1 (n = 26), in which TACE was performed alone, and group 2 (n = 24), in which transcutaneous ablation of HCC with high-intensity focused ultrasound was performed 2-4 weeks after TACE. The tumors were 4-14 cm in diameter (mean, 10.5 cm). Immediate therapeutic effects were assessed at follow-up with Doppler US and computed tomography or magnetic resonance imaging. All patients were followed up for 3-24 months (mean, 8 months) to observe long-term therapeutic effects and complications in both groups. Tumor reduction rates, median survival time, and cumulative survival rates in both groups were calculated by using the unpaired Student t test and Kaplan-Meier method., Results: No severe complication was observed after focused ultrasound ablation, and no unexpected side effects were noted after TACE. Follow-up images showed absence or reduction of blood supply in the lesions after focused ultrasound ablation when compared with blood supply after TACE alone. The median survival time was 11.3 months in group 2 and 4.0 months in group 1 (P = .004). The 6-month survival rate was 80.4%-85.4% in group 2 and 13.2% in group 1 (P = .002), and the 1-year survival rate was 42.9% and 0%, respectively. Median reductions in tumor size as a percentage of initial tumor volume at 1, 3, 6, and 12 months after treatment, respectively, were 28.6%, 35.0%, 50.0%, and 50.0% in group 2 and 4.8%, 7.7%, 10.0%, and 0% in group 1 (P < .01)., Conclusion: The combination of high-intensity focused ultrasound ablation and TACE is a promising approach in patients with advanced-stage HCC, but large-scale randomized clinical trials are necessary for confirmation., ((c) RSNA, 2005.)

Background & Objective: Recently, multi-needles radiofrequency ablation and brachyradiotherapy have been used to treat some solid tumors, such as liver cancer and prostate cancer. This study was to explore the feasibility and efficacy of radiofrequency ablation or ablation combined with 125I seed implantation on treating renal carcinoma., Methods: From Jun. 2000 to Feb. 2004, 11 patients with a total of 13 renal carcinoma lesions were treated with ablation or ablation combined with (125)I seed implantation, including 6 cases of renal carcinoma of solitary kidney, 2 cases of renal carcinoma combined with contralateral renal atrophy resulted from ureteric stone, 1 case of bilateral tumor, and 2 cases with general conditions contraindicated to surgery. Of the 13 lesions, 1 was resected by open operation, 6 were treated with multi-needles radiofrequency ablation, and 6 were treated with ablation and simultaneous (125)I seed implantation around tumor margin., Results: The patients were followed-up for 6-46 months (mean 27 months). One patient died of cardiac muscle infarction 4 months after operation. Of the 10 alive patients, 2 with recurrent tumors 7 months and 13 months after operation were treated with ablation again, 1 with postoperative uremia was treated with intermittent peritoneal dialysis, the rest 7 had normal renal function., Conclusion: Multi-needles radiofrequency ablation or ablation combined with (125)I seed implantation would be an option in the treatment for patients with renal carcinoma of solitary kidney or bilateral kidney lesions, or for patients can't suffer from surgery.

HIV-1 integrase mediates a processed viral DNA into the host genome DNA. The binding modes between HIV-1 integrase and viral/host DNA have not been clarified until now. The C-terminal domain of integrase has been found to be a DNA-binding domain. In this work, we have explored the possible DNA binding sites of dimeric C-terminal domain by docking dinucleotides to HIV-1 integrase. The docking results suggest that two symmetrical DNA-binding sites are likely located on the outside surface of the dimeric C-terminal domain and not located in the groove. Those sites are in agreement with the experimental data.

Objective: A study on the value of antimitochondrial antibody (AMA) and its subtypes anti-M2, anti-M4, and anti-M9 in diagnosing primary biliary cirrhosis (PBC)., Methods: Antimitochondrial antibody was detected by indirect immunofluorescence and anti-M2, anti-M4 and anti-M9 by Western blotting. AMA and anti-M2 of 78 PBC patients, of 35 non-PBC hepato-biliary disease patients and 20 healthy controls were studied and anti-M2, anti-M4 and anti-M9 were studied in 30 of the 78 PBC patients., Results: 96.2% (75/78) of PBC patients were AMA positive and 94.9% (74/78) of PBC patients were anti-M2 positive. Only three among the 35 non-PBC patients were positive for AMA (one with very low titre). None of the 35 non-PBC patients was anti-M2 positive. AMA and anti-M2 were negative in all the healthy controls. Among the 30 anti-M2 positive patients, 16 patients were anti-M4 positive (16/30, 53.3%) and 4 patients were anti-M9 positive (4/30, 13.3%)., Conclusion: AMA and its subtypes (special anti-M2) are important sero-immunological markers for the diagnosis of PBC.

Background: High intensity focused ultrasound (HIFU) is a noninvasive treatment modality that induces complete coagulative necrosis of a deep tumor through the intact skin. The current study was conducted to determine the safety, efficacy, and feasibility of extracorporeal HIFU in the treatment of patients with hepatocellular carcinoma (HCC)., Methods: A total of 55 patients with HCC with cirrhosis were enrolled in this prospective, nonrandomized clinical trial. Among them, 51 patients had unresectable HCC. Tumor size ranged from 4 to 14 cm in diameter with mean diameter of 8.14 cm. According to tumor, node, metastasis (TNM) classification, 15 patients corresponded to stage II, 16 to stage IIIA, and 24 to IIIC. All patients had HIFU, and the median number of HIFU session was 1.69. Safety and efficacy of HIFU were assessed in this trial., Results: No severe side effect was observed in the patients treated with HIFU. Follow-up imaging showed an absence of tumor vascular supply and the shrinkage of treated lesions. Serum alpha-fetoprotein returned to normal level in 34% of patients. The overall survival rates at 6, 12, and 18 months were 86.1%, 61.5%, and 35.3%, respectively. The survival rates were significantly higher in patients in stage II than those in stage IIIA (P = .0132) and in stage IIIC (P = .0265)., Conclusion: As a noninvasive therapy, HIFU appears to be effective, safe, and feasible in the treatment of patients with HCC. It may play an important role in the ablation of large tumors.

Objective: To explore the effects of ureteral stent placement on ureteral peristalsis and the time course of renal pelvic pressure changes., Method: Forty rabbits were randomized equally into 8 groups, including a control group without placement of ureteral stents. The other 7 groups received ureteral stent placement, which were retained for 0, 1, 2, 3, 4, 7, and 12 weeks respectively. When the rabbits were bred for the specified time periods, ureteral peristalsis was observed and renal pelvic pressure measured., Result: The renal pelvic pressure was initially increased after the stent placement, but then followed by gradual decreases. Ureteral peristalsis was not observed after the stent placement., Conclusion: Ureteral stent is a factor for causing obstruction in normal ureter.

T cell-mediated immune responses represent the main cellular antitumor immunity in cancer patients. Recent studies have shown that that both surgical procedure and radiation therapy could cause the functional suppression of lymphocyte-mediated cellular immunity. The purpose of current study is to evaluate whether high intensity focused ultrasound (HIFU) might change a systemic antitumor immunity, particularly T lymphocyte-mediated immunity in cancer patients. A total of 16 patients with solid malignancies were treated with HIFU. Among them, six patients had osteosarcoma (Enneking stage, II(B)4, III(B) 2), five had hepatocellular carcinoma (TNM stage, III 3, IV 2), and five had renal cell carcinoma (TNM stage, III 2, IV 3). Using flow cytometry technique, T lymphocyte and subset, B lymphocyte and natural killer cell (NK) in the peripheral blood were measured in these patients on the day before HIFU and 7 to 10 d after HIFU. The statistical significance of any observed difference is evaluated by Student's t-test. The results showed a significance increase in the population of CD4(+) lymphocytes (p < 0.01) and the ratio of CD4(+) /CD8(+) (p < 0.05) in the circulation of cancer patients after HIFU treatment. The abnormal levels of CD3(+) lymphocytes returned toward the normal range in two patients, CD4(+)/CD8(+) ratio in 3, CD19(+) lymphocytes in one and cytotoxic NK in one, respectively, in comparison to control values. It is concluded that HIFU could enhance a systemic antitumor cellular immunity in addition to local tumor destruction in patients with solid malignancies.

Background & Objective: Protein kinase CK2 is a ubiquitous and pleiotropic Ser/Thr protein kinase in eukaryotic cells. CK2 activity has been shown to be markedly elevated in solid tumors and leukemia cells. Its alpha or alpha' gene is a protooncogene. CK2 is attracting increasing attention as a potential target for anti-neoplastic. This study was to search specific CK2 inhibitors in tumor cells through observation of the inhibitory effects of baicalein on recombinant human protein kinase CK2 holoenzyme and its kinetics in vitro., Methods: Recombinant human protein kinase CK2 alpha' and beta subunits were cloned and expressed by gene engineering, and purified to homogeneous. These 2 subunits were mixed at equal molar ratio to reconstitute CK2 holoenzyme. The CK2 activity was evaluated by detecting radioactivity of 32P of [gamma-32P]ATP which was incorporated into the substrate in various conditions., Results: Baicalein was shown to strongly inhibit the holoenzyme activity of CK2 with IC50 of 2.54 micromol/L. Kinetic studies of baicalein on CK2 showed that baicalein acted as an inhibitor of noncompetitive with ATP(KI=7.73 micromol/L) and mixed types with casein(KI=3.07 micromol/L)., Conclusion: Baicalein is an effective inhibitor of protein kinase CK2 in vitro.

Objective: To investigate the role of bcl-2 and bax expression in apoptosis of hypoxic pulmonary artery smooth muscle cells (PASMCs) of rats induced by Na+/H+ exchanger isoform-1 (NHE-1) inhibition in vitro., Methods: Cultured rat PASMCs transfected with NHE-1 ribozyme gene (PRZ cell), cultured rat PASMCs transfected only with pLXSN vector (PX cell) and non-transfected rat PASMCs (PA cell) were obtained in our previous studies. The cells were cultured under low oxygen tension (<1% O2) for 0, 2, 6, 12, 24 and 48 hours respectively. Then cell apoptosis was examined with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). Intracellular Ca2+([Ca2+]i) of cells was measured using fluorescence dye Fura-2/AM. The changes in mRNA expression of bcl-2 and bax in cells were assessed by semi quantitative reverse transcription-polymerase chain reaction (sqRT-PCR). The expression of Bcl-2 protein and Bax protein of cells was detected immunohistochemically., Results: In contrast to normal PASMCs and cells transfected with pLXSN, the apoptosis rates in recombinant vectors transfected cells increased significantly and continuously. [Ca2+]i values were also increased in these cells, but the increase extent was remarkably lower than in the normal PASMCs (all P<0.01). The expression of bax mRNA and protein of cells transfected with recombinant vectors was increased significantly compared with cells of other two groups. Meanwhile, the expression of bcl-2 mRNA and protein of these cells were decreased significantly., Conclusion: The NHE-1 specific hammerhead ribozymes induce apoptosis of PASMCs under low oxygen tension by inhibiting the increase in intracellular Ca2+ concentration, increasing bax expression and decreasing bcl-2 expression.

Aim: To understand pharmacophore properties of styrylquinoline derivatives and to design inhibitors of HIV-1 integrase., Methods: Comparative molecular field analysis (CoMFA) was performed to analyze three-dimensional quantitative structure-activity relationship (3D-QSAR) of styrylquinoline derivatives. Thirty-eight compounds were randomly divided into a training set of 28 compounds and a test set of 10 compounds. The stability of 3D-QSAR models was proved by the analysis of cross-validated and non-cross-validated methods. Moreover, the binding mode of these compounds and integrase was constructed by AutoDock program., Results: The CoMFA model of the training compounds was reasonably predicted with cross-validated coefficient (q2) and conventional (r2) values (up to 0.696 and 0.754). Then the model was validated by the test set. The resulting CoMFA maps visualized structural requirements for the biological activity of these inhibitors. Docking results showed that a carboxyl group at C-7 and a hydroxyl group at C-8 in the quinoline subunit, bound closely to the divalent metal cofactor (Mg2+) around the integrase catalytic site. Moreover, there is a linear correlation between the binding energy of the inhibitors with integrase and their inhibitory effect., Conclusions: The present study indicated that the CoMFA model together with docking results could give us helpful hints for drug design as well as interpretation of the binding affinity between these inhibitors and integrase.

Objective: To explore the feasibility and efficacy of focused ultrasound for the treatment of vulvar dystrophy, including squamous hyperplasia (SH) and lichen sclerosus (LS)., Methods: A total of 76 eligible patients with vulvar dystrophy (45 SH and 31 LS) were treated with focused ultrasound between 1999 and 2002. Before and after ultrasound therapy, both ultrasonography and biopsies of the lesions were performed to monitor and evaluate the changes of the lesion being treated. The positive expressions of CD34 (cluster of differentiation of endothelial cells), a marker of the epithelial cells of blood vessels and myelin basic protein (MBP), a marker of the oligodendrocytes and Schwann cells were tested using the streptavidin-peroxidase (SP) immunohistochemistry method before and after the ultrasound procedure to evaluate the effects of ultrasound treatment., Results: In two years, follow-up, 49 of 76 cases (32 SH and 17 LS) were cured, 23 (11 SH and 12 LS) improved, and 4 (2 SH and 2 LS) persisted. The response rate was 94.7% (72/76). The positive expression of CD34 and MBP significantly increased at the treated region (P < 0.05). Grey-scale ultrasound imaging showed a localized hypoechoic region after the treatment, which recovered to normal appearance within 7 - 10 days., Conclusion: Vulvar dystrophy can be treated with focused ultrasound effectively and safely. This approach appears to be a new promising treatment method, although further studies are still needed.

We report here the preparation of nickel tetrasulfo-phthalocyanine (NiTSPc)-doped silica xerogel matrix obtained by sol-gel technique. The evolution of silica matrix's texture and the rate during the sol-gel process, as well as silica xerogel's pore structure were studied. Dimerization behavior of NiTSPc molecules was characterized by electronic absorption spectra, and changes in the absorption spectra were correlated with various stages of the sol-gel process. The results show that in the composites NiTSPc concurs in the forms of monomer and dimer, and dimerization is stronger during the latter stages. The reason is given from the change in the micro-environment in which NiTSPc existed during the sol-gel process.

The ideal treatment of localized cancer should directly cause an irreversible and complete death of tumor cells without damage to surrounding normal tissue. High intensity focused ultrasound (HIFU) is such a potential treatment, which induces a complete coagulative necrosis of a tumor at depth through the intact skin. The idea that using an extracorporeal source of therapeutic ultrasound was introduced more than 50 years ago [J. Gen. Physiol. 26 (1942) 179]. However, up to now, most of the studies on HIFU have been dealing with animal experiments because this extracorporeal technique is very complicated in clinical applications. The purpose of this study is to introduce Chinese clinical experience of using extracorporeal HIFU for the treatment of patients with various kinds of solid tumor. From December 1997 to October 2001, a total of 1038 patients with solid tumors underwent HIFU ablation in China. Among them, 313 patients were treated at the Chongqing University of Medical Sciences, China. Pathological examination showed that the target region presented clear evidence of cellular destruction. Small blood vessels less than 2 mm in diameter were severely damaged. Follow-up diagnostic imaging revealed that there was no, or reduced, blood supply, and no uptake of radioisotope in the treated tumor after HIFU, both indicating a positive therapeutic response and an absence of viable tumor. Imaging at 6-12 months showed obvious regression of the lesion. Four-year follow-up data were significantly observed in patients with hepatocellular carcinoma, osteosarcoma, and breast cancer. An extremely low major complication rate was noted. It is concluded that HIFU ablation is a safe, effective, and feasible modality for the ablation of carcinomas.

The theoretical possibility that exposure of a solid malignancy to high-intensity focused ultrasound (US), or HIFU, could lead to an increased rate of metastasis still remains. Using reverse transcriptase polymerase chain reaction (RT-PCR), the potential risk of hematogenous dissemination was assessed in HIFU-treated patients with solid malignancy. RT-PCR can demonstrate the presence or absence of specific RNA fragments. On the day before HIFU ablation, 5-mL peripheral blood samples were collected, and again 5 to 7 days after HIFU, from 26 enrolled patients (hepatocellular carcinoma, HCC: 10; osteosarcoma: 16). Total RNA was isolated and RT-PCR was performed to analyze the mRNA expression of (alpha-fetoprotein (AFP) and bone-specific alkaline phosphatase (BALP) genes. Positive AFP mRNA expression was preoperatively detected in 8 of 10 patients with HCC. In the postoperative specimens, positive expression was also detected in 8 of 10 patients. In 2 patients, circulating tumor cells were found preoperatively, but not postoperatively. Conversely, 2 patients with no circulating tumor cells preoperatively were found to have circulating tumor cells after HIFU. Of 16 osteosarcoma patients, 12 patients had circulating tumor cells and 4 had none. After HIFU treatment, 2 of the 12 patients had converted from presence to absence of circulating cells and the remaining 4 patients remained negative. It is concluded that patients undergoing complete HIFU ablation may demonstrate conversion from presence to absence of circulating tumor-specific marker mRNA, and that HIFU would not enhance the potential risk of metastasis in patients with malignant diseases.

Objective: Using a genotying method which is accurate and rapid to study the molecular epidemiology of EsBLs-producing Escherichia coli in nosocomial infection., Methods: After an analysis of clinical information to ascertain nosocomial infection, EsBLs were detected by phenotype confirmation test and pulsed-field gel electrophoresis (PFGE)., Results: Three EsBLs-producing strains in ICU were verified as epidemiological-related because their dice coefficient index was higher than 95%. The others were verified as non-epidemiological-related since their different lanes were more than 6., Conclusion: PFGE is a perfect molecular typing method to study epidemiolgy and analyze homology.

Objective: To investigate the effect of fragile histidine triad (FHIT) gene on the apoptosis of gastric cancer cells., Methods: FHIT gene was transfected into gastric cancer cell line MGC-803 by liposome. Western blot and immunohistochemical assay were employed to detect the expression of FHIT. Apoptosis and cell cycle were analyzed by flow cytometry (FCM). Morphological changes were observed by light and electron microscope., Results: Stable expression of FHIT protein in transfected MGC-803 cells was obtained. The rates of G0/G1 phase and apoptosis cells were significantly higher in FHIT-transfected MGC-803 cells than those in control cells (64.4%compared with 49.5%and 32.0%compared with 12.2%, respectively)., Conclusion: The results indicate that FHIT gene might be involved in the apoptosis of gastric cancer cells.

The objective of this article is to introduce the early Chinese clinical experience of using extracorporeal focused ultrasound (US) surgery (FUS) for the treatment of solid tumors. From December 1997 to October 2001, a total of 1038 patients with solid tumors underwent FUS ablation in 10 Chinese hospitals. The tumors included primary and metastatic liver cancer, malignant bone tumors, breast cancer, soft tissue sarcomas, kidney cancer, pancreatic cancer, abdominal and pelvic malignant tumors, uterine myoma, benign breast tumors, hepatic hemangioma and other solid tumors. In this article, pathologic changes in tumors treated with FUS, real-time diagnostic imaging for targeting, monitoring and assessment of results by follow-up images are presented. Early clinical results and complications of the technique are also reported.

The complex structures of Human Immunodeficiency Virus Type 1 (HIV-1) integrase binding two highly potent and nontoxic inhibitors, lithospermic acid (M/sub 5/22) and lithospermic acid B (M/sub 5/32), were obtained using docking calculations. Docking results provided detailed information of their binding modes. The binding sites of M/sub 5/22 and M/sub 5/32 were similar to the inhibitor 5-CITEP. The lowest docking energies for HIV-1 integrase binding M/sub 5/22 and M/sub 5/32 are in agreement with their corresponding lower IC/sub 50/ values. Our results on the chemical structure difference between M/sub 5/22 and M/sub 5/32 show that the carboxyl and hydroxyl groups on the side-chain of M/sub 5/32 are important chemical groups which could help to increase the effect against HIV-1 IN replication.

Bioreactor landfills allow a more active landfill management that recognizes the biological, chemical and physical processes involved in a landfill environment. The laboratory-scale simulators of landfill reactors treating municipal solid wastes were studied, the effect of solid waste size, leachate recirculation, nutrient balance, pH value, moisture content and temperature on the rate of municipal solid waste (MSW) biodegradation were determined, and it indicated the optimum pH value, moisture content and temperature decomposing MSW. The results of waste biodegradation were compared with that of the leachate-recirculated landfill simulator and conservative sanitary landfill simulator. In the control experiment the antitheses of a decreasing trend of the organic load, measured as biological oxygen demand and chemical oxygen demand, was shown. An obvious enhancement of effective disposal from conservative sanitary landfill (CSL) simulator, to the leachate-recirculated landfill (LRL) simulator and to the conditioned bioreactor landfill (CBL) simulator would be noted, through displaying the compared results of solid waste settlement, heavy metal concentration in leachate, methane production rate, biogas composition, BOD and COD as well as their ratio.

Background & Objective: It is difficult to stent the obstructed ureters cystoscopically in patients with renal failure caused by radiotherapy for abdominal or pelvic tumors. Commonly draining the kidney with nephrostomy is the only method for the patients. However, external nephrostomy impacts on life quality of the patients. We reported our experience in stenting the obstructed ureters using minimally invasive percutaneous nephrostomy combined with ureteroscopy, and investigated viability of endoscopic treatment for bilateral ureteral strictures following radiotherapy., Methods: Indwelling double pigtail stents in ureter by minimally invasive percutaneous nephrostomy combined with ureteroscopy was carried out in 19 patients with bilateral ureteral strictures following radiotherapy. The clinical outcomes including ureteral drainage and renal function were reviewed., Results: Bilateral retrograde placement of double pigtail stent was performed in 2 cases. Indwelling different size double pigtail stents in unilateral ureters were carried out in 17 cases. The patients were followed-up for 1-24 months. Renal function was improved excellently in 17 cases, with serum creatinine level of 45 micromol/L to 113 micromol/L. Two patients had poor renal function improvement after ureteral stenting and were converted to percutaneous nephrostomy for draining the kidney., Conclusion: Indwelling different size double pigtail stents simultaneously in unilateral ureter by minimally invasive percutaneous nephrostomy is safe, simple, and effective in treating bilateral ureteral obstruction following radiotherapy.

Metathesis reactions between Ru(2)(DMBA)(4)Cl(2) (DMBA = N,N'-dimethylbenzamidinate) and MX (M = Na and K) yielded bis-adduct derivatives Ru(2)(DMBA)(4)X(2) (X = CN (1), N(3) (2), N(CN)(2) (3)). Metathesis reactions between Ru(2)(DMBA)(4)(NO(3))(2) and KI resulted in Ru(2)(DMBA)(4)I(2) (4). Compound 1 is diamagnetic, while compounds 2-4 are paramagnetic (S = 1). Both compounds 1 and 2 undergo two reversible one-electron processes, an oxidation and a reduction, while compound 3 features a quasireversible reduction. Single-crystal X-ray diffraction studies revealed that the Ru-Ru bond lengths are 2.4508(9), 2.3166(7), 2.304[1], and 2.328(1) A for compounds 1-4, respectively. Structural and electrochemical data clearly indicate that the axial ligands impart a significant influence on the electronic structures of diruthenium species.

High-intensity focused ultrasound (HIFU) is a noninvasive treatment that induces complete coagulative necrosis of a tumour at depth through the intact skin. This study was to explore the possibility of using HIFU for the treatment of patients with localised breast cancer in a controlled clinical trial. A total of 48 women with biopsy-proven breast cancer (T(1-2), N(0-2), M0) were randomised to the control group in which modified radical mastectomy was performed, and the HIFU group in which an extracorporeal HIFU ablation of breast cancer was followed by modified radical mastectomy. Short-term follow-up, pathologic and immunohistochemical stains were performed to assess the therapeutic effects on tumour and complications of HIFU. The results showed that no severe side effect was observed in the HIFU-treated patients. Pathologic findings revealed that HIFU-treated tumour cells underwent complete coagulative necrosis, and tumour vascular vessels were severely damaged. Immunohistochemical staining showed that no expression of PCNA, MMP-9, and CD44v6 was detected within the treated tumour cells in the HIFU group, indicating that the treated tumour cells lost the abilities of proliferation, invasion, and metastasis. It is concluded that, as a noninvasive therapy, HIFU could be effective, safe, and feasible in the extracorporeal treatment of localised breast cancer.

Purpose: We present the preliminary results of patients with advanced stage renal malignancy treated with high intensity focused ultrasound (HIFU), and investigate the safety and feasibility of using HIFU in the treatment of selected patients with renal tumors., Materials and Methods: HIFU treatment was performed in 12 patients with advanced stage renal cell carcinoma and 1 patient with colon cancer metastasized to kidney. Patients were followed after treatment to observe complications and long-term therapeutic efficacy. Complications and changes in symptoms seen at presentation were recorded. Mid stream urine specimens were sent for microscopy and serum creatinine was measured postoperatively. Followup radiological examinations were performed to detect tumor response to the ablation., Results: A total of 13 patients received HIFU treatment safely, including 10 who had partial ablation and 3 who had complete tumor ablation. After HIFU hematuria disappeared in 7 of 8 patients and flank pain of presumed malignant origin disappeared in 9 of 10 patients. Postoperative images showed decrease in or absence of tumor blood supply in the treated region and significant shrinkage of the ablated tumor. Of the 13 patients 7 died (median survival 14.1 months, range 2 to 27) and 6 were still alive with median followup of 18.5 months (range 10 to 27)., Conclusions: This preliminary experience suggests that HIFU could be safe and feasible in the treatment of patients with advanced renal malignancy.

Objective: To analyze the relationship of the height difference and bone density (BD) of premenopausal and postmenopausal women., Methods: The height values of 191 premenopausal and postmenopausal women were recorded, and the BD values of lumbar vertebrae and hip were detected by double energy X-ray BD detector., Results: The lower the height of the postmenopausal women, the less the BD value. The BD of lumbar vertebrae dropped 0.025 5 g/cm(2) with each 2 cm of the shortened height, and the BD of hip joint dropped 0.029 2 g/cm(2). The shortened value in postmenopausal women with osteoporosis was statistically greater than that in postmenopausal women without osteoporosis., Conclusion: The BD of the postmenopausal women can be estimated by the calculation of their shortened height value.

Objective: To investigate the expressions of interleukin 6 (IL-6) and Janus kinases (JAKs) in rats with hypoxia induced pulmonary hypertension (HPH)., Methods: Sixty male Wistar rats were randomized into 5 groups with 12 animals in each: a one-week hypoxic group (H(1) group), a two-week hypoxic group (H(2) group), a three-week hypoxic group (H(3) group), a four-week hypoxic group (H(4) group), a normal oxygen group (N group). The rat model of HPH was replicated in normal baric hypoxic cabin. The levels of IL-6 and JAKs mRNA were measured by using reverse transcription-polymerase chain reaction (RT-PCR); The expression of JAKs protein and cell morphologic changes were observed by immunohistochemistry., Results: (1) The levels of IL-6 mRNA in H(1) (1.67 +/- 0.09), H(2) (2.26 +/- 0.12) and H(3) (1.55 +/- 0.11) groups were significantly higher than that in N group (1.20 +/- 0.11, all P < 0.01). The levels of JAK1 mRNA in H(1) (2.11 +/- 0.09), H(2) (2.85 +/- 0.12) and H(3) (2.36 +/- 0.13) groups were significantly higher than that in N group (1.62 +/- 0.10, all P < 0.01); The levels of JAK2 mRNA in H(1) (1.41 +/- 0.07), H(2) (2.02 +/- 0.13) and H(3) (1.36 +/- 0.09) groups were significantly higher than that in N group (1.01 +/- 0.09, all P < 0.01); The levels of JAK3 mRNA in H(1) (0.86 +/- 0.11), H(2) (1.45 +/- 0.10) and H(3) (0.91 +/- 0.13) groups were significantly higher than that in N group (0.55 +/- 0.08, all P < 0.01). The levels of TYK2 mRNA in H(1) (1.36 +/- 0.10), H(2) (1.76 +/- 0.11) groups were significantly higher than that in N group (0.57 +/- 0.07, all P < 0.01). The levels of IL-6 and JAKs mRNA in H(2) group were significantly higher than those in H(1) and H(3) groups (all P < 0.01). (2) Histochemical staining of JAK1 and JAK3 in H(3) group showed yellow cytoplasm of alveolar wall cells, bronchial wall cells and small blood wall cells; The active protein contents of JAK1 (5.36 +/- 0.32) and JAK3 (4.88 +/- 0.29) were markedly increased as compared with N group (1.52 +/- 0.18, 1.22 +/- 0.09, respectively; all P < 0.01) by TIGER image analysis method., Conclusions: The levels of IL-6, JAKs mRNA and protein can increase in pulmonary tissues of rats with HPH. It suggests that IL-6/JAKs pathway may take part in the pathogenesis of HPH.

The products of the reaction between CN(-) and four different diruthenium complexes of the type Ru(2)(L)(4)Cl where L = 2-CH(3)ap (2-(2-methylanilino)pyridinate anion), ap (2-anilinopyridinate anion), 2-Fap (2-(2-fluoroanilino)pyridinate anion), or 2,4,6-F(3)ap (2-(2,4,6-trifluoroanilino)pyridinate anion) are reported. Mono- and/or dicyano adducts of the type Ru(2)(L)(4)(CN) and Ru(2)(L)(4)(CN)(2) are found exclusively as reaction products when either the 2-CH(3)ap or the ap derivative is reacted with CN(-), but diruthenium complexes with formulations of the type Ru(2)(F(x)ap)(3)[mu-(o-NC)F(x-1)ap](mu-CN) or Ru(2)(F(x)ap)(4)(mu-CN)(2) (x = 1 or 3) are also generated when Ru(2)(Fap)(4)Cl or Ru(2)(F(3)ap)(4)Cl is reacted with CN(-). More specifically, four products formulated as Ru(2)(Fap)(4)(CN), Ru(2)(Fap)(4)(CN)(2), Ru(2)(Fap)(3)[mu-(o-NC)ap](mu-CN), and Ru(2)(Fap)(4)(mu-CN)(2) can be isolated from a reaction of CN(-) with the Fap derivative, but the exact type and yield of these compounds depend on the temperature at which the experiment is carried out. In the case of the F(3)ap derivative, the only diruthenium complex isolated from the reaction mixture has the formulation Ru(2)(F(3)ap)(3)[mu-(o-NC)F(2)ap](mu-CN) and this compound has structural, electrochemical, and spectroscopic properties quite similar to that of previously characterized Ru(2)(F(5)ap)[mu-(o-NC)F(4)ap](mu-CN). Both the mono- and dicyano derivatives synthesized in this study possess the isomer type of their parent chloro complexes. The Ru-Ru bond lengths of Ru(2)(ap)(4)(CN) and Ru(2)(2-CH(3)ap)(4)(CN) are longer than those of Ru(2)(ap)(4)Cl and Ru(2)(CH(3)ap)(4)Cl, respectively, and this is accounted for by the strong sigma-donor properties of the CN(-) ligand as compared to Cl(-). The Ru-C bonds in Ru(2)(ap)(4)(CN)(2) are significantly shorter than those in Ru(2)(ap)(4)(CN), thus revealing a greatly enhanced Ru-CN interaction in the dicyano adduct, a result which is also indicated by the fact that nu(CN) in Ru(2)(ap)(4)(CN)(2) is 50 cm(-1) higher than nu(CN) in Ru(2)(ap)(4)(CN). Although both (4,0) Ru(2)(ap)(4)(CN)(2) and (3,1) Ru(2)(Fap)(4)(CN)(2) possess the same formulation, there are clear structural differences between the two complexes and this can be explained by the fact that the two cyano derivatives possess a different binding symmetry of the bridging ligands. Each mono- and dicyano adduct was electrochemically investigated in CH(2)Cl(2) containing TBAP as supporting electrolyte. Ru(2)(ap)(4)(CN), Ru(2)(CH(3)ap)(4)(CN), and Ru(2)(Fap)(4)(CN) undergo one reduction and two oxidations. The two dicyano adducts of the ap and Fap derivatives are characterized by two reductions and one oxidation. The potentials of these processes are all negatively shifted in potential by 400-720 mV with respect to half-wave potentials for the same redox couples of the monocyano derivatives, with the exact value depending upon the specific redox reaction.

Proliferation, invasion, immortalization and metastasis are the main malignant characteristics of cancer. Previous studies have shown that high-intensity focused ultrasound (US), or HIFU, can induce irreversible damage both to breast cancer cells and to tumor blood vessels. However, light microscopy alone may not always show this clearly. In this study, molecular biologic techniques were used to examine any changes in molecular markers associated with malignant behavior after exposure to HIFU. A total of 48 women with breast cancer were randomized to a control group (mastectomy) and a HIFU group (HIFU followed by mastectomy). Immunohistochemical staining, messenger RNA (mRNA) in situ hybridization and telomere-repeat amplification protocol-enzyme-linked immunosorbent assay (TRAP-ELISA) techniques were used to detect tumor expression of proliferating cell nuclear antigen (PCNA), cell adhesion molecule CD44v6, matrix metalloproteinase-9 (MMP-9), erbB2 mRNA, and to measure telomerase activity in both groups. The results demonstrated that there were significant alterations in expression of PCNA, CD44v6, MMP-9, erbB2 mRNA, and a dramatic decrease in telomerase activity in the HIFU group. It is concluded that malignant tumor characteristics are arrested by HIFU, and that biologic factors are potential markers for assessing HIFU efficacy.

An effective and fast minimization approach is proposed for the prediction of protein folding, in which the 'relative entropy' is used as a minimization function and the off-lattice model is used. In this approach, we only use the information of distances between the consecutive Calpha atoms along the peptide chain and a generalized form of the contact potential for 20 types of amino acids. Tests of the algorithm are performed on the real proteins. The root mean square deviations of the structures of eight folded target proteins versus the native structures are in a reasonable range. In principle, this method is an improvement on the energy minimization approach.

An efficient soft docking algorithm is described for predicting the mode of binding between an antibody and its antigen based on the three-dimensional structures of the molecules. The basic tools are the "simplified protein" model and the docking algorithm of Wodak and Janin. The side-chain flexibility of Arg, Lys, Asp, Glu, and Met residues on the protein surface is taken into account. A combined filtering technique is used to select candidate binding modes. After energy minimization, we calculate a scoring function, which includes electrostatic and desolvation energy terms. This procedure was applied to targets 04, 05, and 06 of CAPRI, which are complexes of three different camelid antibody VHH variable domains with pig alpha-amylase. For target 06, two native-like structures with a root-mean-square deviation < 4.0 A relative to the X-ray structure were found within the five top ranking structures. For targets 04 and 05, our procedure produced models where more than half of the antigen residues forming the epitope were correctly predicted, albeit with a wrong VHH domain orientation. Thus, our soft docking algorithm is a promising tool for predicting antibody-antigen recognition., (Copyright 2003 Wiley-Liss, Inc.)

Objective: To study high intensity focused ultrasound (HIFU) therapy combined with transcatheter arterial chemoembolization (TACE) in the treatment of patients with advanced hepatocellular carcinoma (HCC)., Methods: Fifty patients with unresectable HCC (TNM stage IV) were randomized into a TACE (T) group and a TACE plus HIFU (T + H) group. Twenty-six patients underwent TACE alone, and 24 HIFU ablation 2 - 3 weeks after TACE. The mean follow-up time for all patients was 8.16 +/- 2.79 months (range 3 to 24 months). The median survival, 6-month-, 1-year survival rates and average survival of patients who died were calculated by Kaplan-Meier method and Fisher exact test., Results: The median survival time, 6-month and 1-year survival rates were 11.3 months, 80.4 - 85.4% and 42.9% in T + H group, in contrast to 4 months, 13.2% and 0% in T group with significant differences (P < 0.01). The average survival time of patients who died was 10.21 +/- 4.12 months in T + H group, as compared with 4.35 +/- 2.39 months in T group also with significant differences (P < 0.01)., Conclusion: High intensity focused ultrasound therapy (HIFU) combined with transcatheter arterial chemoembolization (TACE), being better than TACE alone, may become one of the most effective treatments for patients with unresectable HCC.

Objective: To explore the proliferation of primary cultured rats hepatocytes after delivery of exogenous hepatocyte growth factor (HGF) gene which was inserted into the genome of replication-deficient recombinant adenovirus vector., Methods: The recombinant adenovirus-AdHGF which could express HGF was generated by homologous recombination. After the HGF gene was delivered into the hepatocytes, the expression of both HGF and c-met/HGF receptor mRNA in the cells was detected by RT-PCR and the level of HGF in the culture supernatant was also assayed by ELISA. On the other hand, cell proliferation was compared between before and after delivery of the HGF gene by MTS assay and the percentages of cell cycles were analyzed by flow cytometry. In addition, the expression of proliferating cell nuclear antigen (PCNA) was determined by immunocytofluorescent stain., Results: 4 x 10(10) efu/ml titer of AdHGF was obtained after recombination, RT-PCR indicated that the expression of HGF mRNA in hepatocytes increased on the third day after infected by the viruses and c-met/HGF receptor mRNA was also up-regulated. The HGF level in the culture supernatant assayed by ELISA was (5,939.0+/-414.39) pg/ml, which was much higher than that in the control (208.1pg/ml+/-37.20pg/ml, F=13.661, P<0.01). In addition, the proliferation of hepatocytes infected with AdHGF increased significantly according to MTS method (F>or=15.158, P<0.01) and more hepatocytes in G0/G1 stages changed into S stage (chi2=41.616, P<0.01), accordingly, PCNA index increased from 6.42+/- 1.88 to 14.56+/-2.85 (F=42.122, P<0.01)., The Results: show that HGF gene delivered into hepatocytes by AdHGF can be expressed with high efficiency in the cells, which can stimulate hepatocytes proliferation. It may be an effective tool for hepatocyte transplantation by gene modified donor hepatocytes.

Objective: To evaluate the potential preventive effect of Sophoricoside on bone loss in ovariectomized rats., Methods: Female SD rats (n=50, 6 months old) were either sham-operated (SHAM group, n=10) or ovariectomized (n=40). Three days after operation, ovariectomized rats were randomly assigned to groups as follows: 10 received Sophoricoside 4 mg.kg(-1).d(-1) (SL group), 10 received Sophoricoside 8 mg.kg(-1).d(-1) (SM group), 10 received Sophoricoside 16 mg.kg(-1).d(-1) (SH group) and 10 were untreated (OVX group). After Sophoricoside were given orally for one month, the histomorphometric parameters in the secondary spongiosa of proximal tibia and lumbar vertebrae were examined., Results: Compared with OVX group, SM and SH two kinds of treatment caused 15.28% and 22.81% increment in percent trabecular area (BV/TV, %) in the secondary spongiosa of proximal tibia, 14.23% and 21.2% increment in lumbar vertebrae. Accordingly in these two groups, there was a significant decrease in trabecular separation (FLAW, microm), but increment in trabecular width (Tb. Th, microm) and conjunction points (JOINT). But in SL group, the preventive effect was not observed., Conclusion: Sophoricoside can be efficient in preventing ovariectomy-induced bone loss in rats.

An efficient 'soft docking' algorithm is described to assist the prediction of protein-protein association using three-dimensional structures of molecules. The basic tools are the 'simplified protein' model and the docking algorithm of Wodak and Janin. The side chain flexibility of Arg, Lys, Asp, Glu and Met residues at the protein surface is taken into account. The complex type-dependent filtering technique on the basis of the geometric matching, hydrophobicity and electrostatic complementarity is used to select candidate binding modes. Subsequently, we calculate a scoring function which includes electrostatic and desolvation energy terms. In the 44 complexes tested including enzyme-inhibitor, antibody-antigen and other complexes, native-like structures were all found, of which 30 were ranked in the top 20. Thus, our soft docking algorithm has the potential to predict protein-protein recognition.

An efficient soft docking algorithm is described to predict the mode of binding between two proteins based on the three-dimensional structures of molecules. The molecular model used in this work was grounded on the "simplified protein" model used in Janin's docking algorithm. The side chain flexibility of the amino acid residues Arg, Lys, Asp, Glu and Met at the protein surface was considered through softening the molecular surface. A double filtering technique was used to eliminate most of the unlike binding modes. The energy minimization was performed on the retained structures, and then these structures were evaluated with the scoring function which included electrostatic, desolvation and van der Waals energy terms. The 26 complexes were used to test this docking algorithm and good results were obtained. The native-like conformations of all the complexes were all found, of which 20 were ranked in the top 10.

The electrostatic force including the intramolecular Coulombic interactions and the electrostatic contribution of solvation effect were entirely calculated by using the finite difference Poisson-Boltzmann method (FDPB), which was incorporated into the GROMOS96 force field to complete a new finite difference stochastic dynamics procedure (FDSD). Simulations were performed on an insulin dimer. Different relative dielectric constants were successively assigned to the protein interior; a value of 17 was selected as optimal for our system. The simulation data were analyzed and compared with those obtained from 500-ps molecular dynamics (MD) simulation with explicit water and a 500-ps conventional stochastic dynamics (SD) simulation without the mean solvent force. The results indicate that the FDSD method with GROMOS96 force field is suitable to study the dynamics and structure of proteins in solution if used with the optimal protein dielectric constant., (Copyright 2002 Wiley-Liss, Inc.)

Three useful variables from the interfaces of 20 protein-protein complexes were investigated. These variables are the side-chain accessible number (N(b)), the number of hydrophilic pairs (N(pair)) and buried a polar solvent accessible surface areas (DeltaDeltaASA(apol)). An empirical model based on the three variables was developed to describe the free energy of protein associations. As the results show, the side-chain accessible numbers characterize the loss of side-chain conformational entropy of protein interactions and the effective empirical function presented here has great capability for estimating the binding free energy. It was found that the variables of interface information capture most of the significant features of protein-protein association. Also, we applied the model based on the variables as a rescoring function to docking simulations and found that it has the potential to distinguish the 'true' binding mode. It is clear that the simple and empirical scale developed here is an attractive target function for calculating binding free energy for various biological processes to rational protein design.

The purpose of this study was to explore the sequential imaging and histologic alterations of tumor blood vessels in the patient with solid malignancies after extracorporeal treatment of high-intensity focused ultrasound (HIFU). A total of 164 patients underwent extracorporeal HIFU ablation of malignant solid tumors. After HIFU treatment, enhanced magnetic resonance imaging (MRI), color Doppler ultrasound (US) imaging, dynamic radionuclide scanning, digital subtraction angiography, and histologic study were performed to monitor the response of tumor vessels to HIFU ablation. Compared with tumor images in the patients before HIFU, clinical images showed an abrupt interruption, followed by the cessation of blood flow within the tumor vessels after HIFU treatment. The histologic examination indicated that not only the treated tumor cells showed coagulative necrosis, but also small tumor vessels were severely damaged by the HIFU treatment. The results strongly imply that the damaged tumor vessels might play a critical role in secondary tumor cell death, and then indirectly strengthen the destructive force of focused US beams on tumor tissue. It is concluded that tumor vessel damage can be induced by HIFU, which may be a promising strategy in the treatment of patients with solid malignancies.

Aim: To observe the killing effects of ganciclovir (GCV) on the human pulmonary adenocarcinoma cell A549 transduced with Herpes simplex virus I type thymine kinase (HSV1-TK) gene in vitro and in vivo., Methods: A retroviral vector containing the TK gene was constructed and transduced into a pulmonary carcinoma cell A549 by electroporation, to observe the sensitivity of the transfected cell to GCV in vitro and the bystander effect (MTT assay). Tumor cell apoptosis caused by the TK/GCV system was observed with a flow cell meter (FCM) and a scan electronic microscope (SEM). Recombination and expression of the TK gene were examined with DNA PCR and in situ hybridization, respectively. The therapeutic effect of GCV on subcutaneous tumor growth between transfected and parental cells was also compared., Results: The sensitivity of the transfected cell to GCV was 46 times higher than that of the parental cell, and the bystander effect was stronger in high cell density than in low cell density. The subG0G1 peak was shown on the DNA histogram after A549-Tk cell was treated with 50 micromol/L GCV for 3 days by FCM, but not in the A549 cell. A cell cycle analysis showed that the apoptotic cell in the A549-TK and A549 cells were (12.2+/-1.7) % and (1.3 +/- 0.3) %, respectively (P < 0.01). The cell apoptosis features of nuclear condensation, apoptotic vesicle, and nuclear showing semimoon feature were found in the A549-TK cell by SEM, but not in the A549 cell. Recombination and expression of the TK gene were positive in the transfected cell. In vivo, the growth of tumors formed by the transfected cell was apparently inhibited by GCV, but not in the control group., Conclusion: The transfected cell obtained sensitivity to GCV and the bystander effect was closely related to intercellular touch. The TK/GCV system killing tumor cell was related to cell apoptosis. GCV inhibited the growth of tumors which were inoculated by A549-TK cell in vivo.

The purpose of this study was to investigate the pathologic changes of extracorporeal ablation of human malignant tumors with high-intensity focused ultrasound (HIFU). HIFU treatment was performed in the 164 patients with liver cancer, breast cancer, malignant bone tumor, soft tissue sarcoma and other malignant tumors at focal peak intensities from 5000 W x cm(-2) to 20,000 W x cm(-2), with operating frequencies of 0.8 to 3.2 MHz. To explore the pathologic impact of extracorporeal HIFU, 30 patients with malignant carcinoma underwent surgical removal after HIFU treatment. Pathologic findings showed that the treated tissues demonstrated homogeneous coagulative necrosis with an irreversible tumor cell death and severe damage to tumor blood vessels at the level of microsvasculature within the HIFU-targeted region. Thermolesions to intervening tissue were never observed. The treated region had a sharp border comprising only several cell layers between the treated and untreated areas. The repair of lesions had the processes of necrotic tissue absorption and granulation tissue replacement. It is concluded that extracorporeal treatment of human solid malignancies with HIFU could be safe, effective and feasible. As a noninvasive therapy, HIFU would be used clinically to treat patients with solid malignancies.

We present the design and fabrication of a phenanthrenequinone-doped poly(methyl methacrylate) photopolymer material. Large blocks of samples were made, and the material showed negligible shrinkage after optical exposures. We recorded and reconstructed 250 holograms at a single spot, using a 1-cm(3) block.

To investigate the process of ion permeation in an ion channel systematically, we performed molecular dynamics (MD) simulations on a gramicidin A (GA)-phospholipid model system with an ion in the channel pore region. Each of the three types of ions (Ca2+, Na+ Cl-) was placed at five different positions along the channel axis by replacing a water molecule. MD simulations were performed on each system at constant pressure and constant temperature. The MD trajectories showed that the Ca2+ and Na+ ions could stably fluctuate in the pore region, but the Cl- ion was pushed out because of the unfavorable interaction with the channel. This result is consistent with experimental data. It was also found that the conformation of the GA channel underwent a significant change due to the presence of the ion, and the two ends of the GA monomer were more flexible than its middle region. In particular, the dramatic change of local pore radius near the ion indicated this kind of deformation. The strong interaction between the ion and carbonyl oxygen atoms of GA was the major contributor to this change. Furthermore, it was found that the ethanolamine group of the GA molecule was the most flexible group in the GA channel and often observed to block the entrance of GA. These results imply that the deformation of channel structure plays a very important factor in ion permeation, and the ethanolamine group may play a key role in regulating ion entry into the pore. In conclusion, our results indicate that the ion has a dominant influence on the structure of the GA channel and that the flexibility of the ion channel is a crucial factor in the ion permeation process.

As a continuing study of chemical characterization of crude drug processing, we have been analyzing the chemical constituents in licorice roots before and after processing. At first, we analyzed chemical constituents in licorice roots of various origins. Next, we have developed the HPLC analytical method, by which saponins and flavonoids, major constituents in various licorice roots, were determined simultaneously in a quantitative manner. In this paper, by means of the HPLC analytical method, chemical constituents in licorice roots, processed and unprocessed, were determined. It was found that nonglycosidic flavonoid constituents were mostly lost while root bark removing, whereas, in roasted licorice roots, sugar chains in the saponin and glycosidic flavonoid constituents were hydrolyzed stepwise during roasting through hydrothermolysis.

Excessive production of nitric oxide (NO) may have cytotoxic effects through the formation of peroxynitrite with superoxide. The extract of Ginkgo biloba leaves (EGb) has been demonstrated to be a potent scavenger of free radicals. Although EGb has been shown recently to inhibit NO production in macrophages, its effect on NO production in endothelial cells is largely unknown. The objective of this study was to elucidate the mechanism by which EGb affects NO production in a human endothelial cell line (ECV304). After cells were incubated with EGb (10-100 microg/mL) for 2 or 4 hr, the amounts of NO metabolites released by the cells were quantitated, and cellular NOS activities were determined following the conversion of [3H]arginine to [3H]citrulline. NOS protein expression was determined by western immunoblotting analysis. mRNA levels were examined by reverse transcription-polymerase chain reaction (RT-PCR) analysis. EGb (50 microg/mL) caused a 30% reduction of NO metabolites released by endothelial cells. Following EGb treatment, cellular inducible NO synthase (iNOS) activity was reduced by 28% with a concomitant reduction in the levels of iNOS protein mass and mRNA. There was no change in the activity or protein mass of constitutive NO synthase in these cells. EGb inhibited NO production by attenuating the level of iNOS mRNA in ECV304 cells. Selective inhibition of iNOS by EGb may be therapeutically relevant in modulating NO production in endothelial cells.

A method for constructing the suitable initial configuration of the membrane-protein system for molecular dynamics (MD) simulations is presented. This method could provide some hydrated initial configurations and help us to determine the best surface area of the system by contracting the surface area and comparing the optimized lowest energy of the system by energy minimization. The gramicidin A (GA) channel in;the fully hydrated dimyristoylphosphatidylcholine (DMPC) bilayer was used as our model. Three configurations with different surface areas were selected and applied for one 400 ps and two 300 ps MD simulations at constant pressure and temperature. All simulations were fairly stable without any constraints. Through analysis of the MD trajectories we found that the system with the best surface area was more stable than the other two systems, whose sizes were changed in the simulations. Further analysis of the bilayer normal length and the order parameters of the lipid alkyl tails indicates that the system with the best surface area shows some characteristics of the L(alpha) phase, while both the smaller and the larger size systems have distinct deviations from the L(alpha) phase that we expect. This illustrates that the correct surface area and the suitable initial configuration have an important influence on the phase of the membrane in the MD simulation. In addition, by comparing the root mean square differences of GA relative to the initial structure and interaction energy between different components of the system for all three systems, we find that the state of the DMPC bilayer has exerted a significant influence on the structure of GA. All these results demonstrate the validity of our method for constructing the initial configuration of the membrane-protein system for MD simulations.