Hydrogel-based co-delivery of CIK cells and oncolytic adenovirus armed with IL12 and IL15 for cancer immunotherapy.

Intratumoral injection of various effector cells combined with oncolytic adenovirus expressing antitumor cytokines exert an effective antitumor immune effect by oncolysis and altering the tumor microenvironment. However, this combination therapy had certain limitations. When used in high concentrations, effector cells and oncolytic viruses can spread rapidly to surrounding non-target tissues. And because both therapies used in combination are immunogenic and exhibit shorter biological activity, multiple injections were required to attain an adequate therapeutic index. To overcome these drawbacks, we encapsulated gelatin-based hydrogel capable of co-deliver oncolytic adenovirus armed with IL12 and IL15 (CRAd-IL12-IL15) and CIK cells for enhancing and prolonging the antitumor effects of both therapies after a single intratumoral injection. The injectable and biodegradable hydrogel reduced the dispersion of high-dose oncolytic adenovirus and CIK cells from the injection site to the liver and other non-target tissues. In this study, a novel oncolytic adenoviral vector CRAd-IL12-IL15 was constructed to verify the cytokine expression and oncolytic ability, which can upregulate the expression levels of Bcl-2, Cish and Gzmb in tumor cells. The CRAd-IL12-IL15 + CIKs/gelatin treatment maintained sustained release of CRAd-IL12-IL15 and active CIK cells over a longer period of time, attenuating the antiviral immune response against adenovirus. In conclusion, the results suggested that hydrogel-mediated co-delivery of CRAd-IL12-IL15 and CIK cells might be a an approach to overcome limitations. Both treatments could be effectively retained in tumor tissue and sustained to induce potent anti-tumor immune responses with a single administration.

Novel hGHRH homodimer promotes fertility of female infertile hamster by up-regulating ovarian GHRH receptor without triggering GH secretion.

Extra-hypothalamic growth hormone-releasing hormone (GHRH) plays an important role in infertility. The female infertility models were formed by intraperitoneally injecting cyclophosphamide in 5-week-old Chinese hamster once in a week for 5 weeks. All the models mated with healthy male hamster in the ratio of 1:1 in the experimental 6-8th week and the couples were separated to breed in the 9-10th week. 20 mg/kg of cyclophosphamide induced temporary interference of reproduction and did not cause significant difference in the weight of body, bilateral ovaries, or liver. By intramuscularly injecting twice in a week during the experimental 4-10th week, 2, 4, 8 mg/kg of Grin induced 30, 42.9, 60% of total pregnancy rates in a dose-dependent manner whereas 200 U/kg of hMG induced 50% of total pregnancy rates. The single cyclophosphamide dose caused strongly eosinophilic ovarian cells, scattered early follicles, many atretic follicles, and no corpora luteum was observed. The hMG group individually presents many follicles at all levels, especially secondary ones in the ovarian cortex and medulla. Much of loose connective tissue, vacuoles, and sparse interstitial cells distribute in the medulla. Grin induced many follicles at all dose levels and corpora lutea in the cortex, and the compactly aligned interstitial cells occurred in the whole ovarian tissue. The less TUNEL staining and higher expression of ki67 showed the proliferation and protection effect of Grin on ovarian cells. Grin obviously promotes fertility by up-regulating ovarian GHRH receptor and strengthening the development and maturation of follicles without triggering central and ovarian GH secretion.

Diterpenoid B derived from Plectranthus excisus inhibits the melanoma cell cycle in the B16 melanoma cell line.

Plectranthus excisus is widely distributed throughout northeast China. Its active ingredient, diterpenoids, exhibits significant antitumor effects. The present study examined the antitumor effects of diterpenoid B (DB), derived from Plectranthus excisus, and demonstrated that DB inhibited the proliferation of tumor cells by inhibiting the cell cycle. Reverse transcription­quantitative polymerase chain reaction and western blot analysis were used to determine mRNA and protein expression levels, respectively. The results revealed that exposure to DB increased the expression levels of the transformation associated, protein 53, and cyclin­dependent kinase inhibitor 1A, and decreased the expression of cyclin­dependent kinase 2. The results of the present study demonstrated that DB can inhibit cell cycle progression and, therefore, offers potential as a beneficial antitumor drug.

Exfoliated layered copper phosphonate showing enhanced adsorption capability towards Pb ions.

Copper 5-(2-bromothienyl)phosphonate (1) with a layered structure is obtained via solvothermal reaction. The layers can be successfully exfoliated using a "top-down" approach, resulting in 2D nanosheets. The exfoliated sample shows an enhanced adsorption capability to the Pb(II) ions in aqueous solution compared with that of the bulk material.

Enhanced magnetic hardness in a nanoscale metal-organic hybrid ferrimagnet.

A new layered metal-organic hybrid compound, namely, [Co(3)(µ(3 -OH)(2)(BTP)(2)] (1; BTP=4-(3-bromothienyl)phosphonate), is reported. The inorganic layer can be viewed as a pseudo-Kagomé lattice composed of corner-sharing irregular triangles of Co(3) (µ(3)-OH), with the cavities filled with the PO(3) groups. The interlayer space is occupied by the 3-bromothienyl groups of BTP(2-). The bulk sample of compound 1 experiences a long-range ferromagnetic ordering below 30.5 K, with a coercivity (H(c)) of 5.04 kOe at 5 K. A systematic study on the size-dependent magnetic coercivity of 1 reveals that the coercivity of 1 increases with reduced particle size from the micrometer to the nanometer scale. When the particle size is about 50-200 nm, the coercivity reaches 24.2 kOe at 5 K. The results demonstrate that compound 1 can vary from a soft magnet to one of the hardest molecule-based magnets, simply by reducing the particle size to nanoscale region.

Silver(I) pyrophosphonates: structural, photoluminescent and thermal expansion studies.

The solvothermal reactions of silver(I) salts with mono-organophosphonic acids, i.e. 3-thienylphosphonic acid (3-TPA), phenylphosphonic acid (PPA), α-naphthylphosphonic acid (α-NPA) and cyclohexylphosphonic acid (CPA), yield four new silver(I) pyrophosphonates, namely: [Ag(2)(ptp)] (1), [Ag(2)(ppp)] (2), [Ag(3)(CH(3)CN)(pnp)(pnpH)] (3), and [Ag(3)(pcp)(pcpH)] (4) [ptp(2-) = pyro-3-thienylphosphonate, ppp(2-) = pyrophenylphosphonate, pnp(2-) = pyro-α-naphthylphosphonate, pcp(2-) = pyrocyclohexylphosphoante]. In all cases, the pyrophosphonate ligands are generated in situ from their relative mono-organophosphonic acids, mediated by silver(I) ions. Single crystal structural determinations reveal that compounds 1 and 2 display two-dimensional layer architectures, while 3 and 4 show one-dimensional chain structures. Structure 1 can be best described as a layer made up of Ag(4)O(P)(6) clusters linked by O-P-O units and AgAg contacts, with the organic groups grafted on the two sides of the inorganic layer. A similar layer structure is found in 2 except that the AgAg interactions are absent. Compound 3 shows a chain structure where the silver ions are bridged by the phosphonate oxygen atoms forming an infinite Ag-O(P) chain which is decorated by the pyrophosphonate ligand and CH(3)CN. Compound 4 has another type of chain structure made up of Ag-O(P) with extensive Ag···Ag argentophilic interactions. Solid state photoluminescent properties and thermal expansion behaviors are also investigated.

Enhanced electrochemiluminescence efficiency of Ru(II) derivative covalently linked carbon nanotubes hybrid.

The synthesized derivative Ru(bpy)(3) covalently linked CNTs hybrid shows good electrochemical activity and ca. 17 times higher luminescence quantum efficiency than the adsorbed derivative Ru(bpy)(3). The Ru-CNTs based ECL sensor exhibits high stability toward determination of TPA with a detection limit as low as 8.75 pM.

Layered copper compounds based on 4-(3-bromothienyl)phosphonate (BTP): weak ferromagnetism observed in [Cu2(4,4'-bpy)0.5(BTP)2].H2O.

Two copper phosphonates based on 4-(3-bromothienyl)phosphonic acid (BTPH2) with formula Cu(H2O)(BTP) (1) and [Cu2(4,4'-bpy)0.5(BTP)2].H2O (2) are reported. Compound 1 has a layered structure in which dimers of {Cu2O2}, made up of edge-sharing {CuO6} octahedra, are bridged by both the {PO3C} tetrahedra through corner-sharing and the water molecules. Compound 2 exhibits a novel layered structure where the {Cu2(BTP)2} columns, containing {Cu4O6} tetrameric cores and O-P-O linkages, are connected by 4,4'-bipyridine. In both cases, the 3-bromothienyl groups are grafted on the two sides of the layers. Magnetic studies reveal that antiferromagnetic interactions are mediated between the Cu(II) centers in 1, while for 2, weak ferromagnetism resulted from spin canting is observed. Electrochemical properties of compounds 1-2 are also investigated.

Ag(I)-mediated formation of pyrophosphonate coupled with C-C bond cleavage of acetonitrile.

A general approach to obtain pyrophosphonates is achieved by reacting arylphosphonic acids with AgNO(3) in CH(3)CN under solvothermal conditions; the process is coupled with the C-C bond cleavage of acetonitrile and the formation of novel complexes [Ag(n)(RPO(2)(O)O(2)PR)(m)](CN); the mechanism of the process is proposed.

Direct electrochemistry and electrocatalysis of hemoglobin at three-dimensional gold film electrode modified with self-assembled monolayers of 3-mercaptopropylphosphonic acid.

Multilayered hemoglobin (Hb) molecules were successfully immobilized on three-dimensional gold film electrode modified with self-assembled monolayers (SAMs) of 3-mercaptopropylphosphonic acid. Direct electrochemistry of the immobilized multilayered Hb occurs with high thermal stability and electrochemical stability. In the multilayered Hb film, the most inner Hb molecules can directly transfer electron with the electrode, while the Hb protein beyond this layer communicates electron with the electrode via protein-protein electron exchange. In addition, the proposed functional interface can greatly enhance electron transfer rate of the immobilized Hb protein (k(s) = 15.8 +/- 2.0 s(-1)) due to the increase of roughness of the gold substrate. Under optimized experimental conditions, the multilayered Hb film displays good bioelectrocatalytic activity toward the reduction of hydrogen peroxide. This electrochemical sensor shows fast response (less than 1 s), wide linear range (7.8 x 10(-8) to 9.1 x 10(-5) M) and low detection limit (2.5 x 10(-8) M), which can be attributed to good mass transport, large Hb proteins loading per unit area and fast electron transfer rate of Hb protein.

3-Mercaptopropylphosphonic acid modified gold electrode for electrochemical detection of dopamine.

It is reported for the first time that the phosphonic acid terminated self-assembled layers assembled on a gold surface could be used as a functional interface to sensitively and selectively detect dopamine (DA) in the presence of highly concentrated ascorbic acid (AA). X-ray photoelectron spectroscopic (XPS) and reductive desorption measurements proved that the 3-mercaptopropylphosphonic acid [HS-(CH(2))(3)-PO(3)H(2), MPPA] was successfully immobilized on the gold surface via S-Au bond. Making use of the electrostatic interactions between the negatively charged phosphonic acid groups on the electrode surface and DA and AA, the MPPA modified electrode was successfully used to electrochemically discriminate between DA and AA. Cyclic voltammetry measurements shown that the MPPA modified electrodes almost completely inhibited the electrochemical reaction of AA. Therefore, DA can be selectively and sensitively determined by using the differential pulse voltammetry technique although high concentration of AA is present.

Direct electrochemistry of cytochrome c on a phosphonic acid terminated self-assembled monolayers.

The direct electrochemistry of cytochrome c (cyt c) on a gold electrode modified with 3-mercaptopropylphosphonic acid [HS-(CH(2))(3)-PO(3)H(2), MPPA] self-assembled monolayers (SAMs) was for the first time investigated. Electrochemical measurements and surface-enhanced infrared absorption spectroscopic reveal that the adsorption kinetics of cyt c on the MPPA-SAMs is very fast (saturation adsorption is completed within 5s) and the immobilized cyt c molecules retain their native secondary protein structure. The nature of interaction between cyt c and -PO(3)H(2) groups is mainly the electrostatic interaction. The direct electrochemistry of the immobilized cyt c on the -PO(3)H(2) terminated SAMs with short chain is nearly reversible. Its formal potential (E(0)'=18+/-3 mV vs. SCE) is very close to that of cyt c in an aqueous solution (E(0)'=18-22 mV vs. SCE). In addition, the electron transfer rate of cyt c immobilized on -PO(3)H(2) terminated SAMs is relatively slow as compared to -SO(3)H and -COOH terminated SAMs, indicating excess negative charge density on the SAMs surface will decrease the electron transfer rate of cyt c.

Diagnostic strategies and the incidence of prostate cancer: reasons for the low reported incidence of prostate cancer in China.

We have analysed the reasons for the low reported incidence of prostate cancer in China and argue for early diagnosis and treatment of this disease. According to the 2002 database of the International Agency for Research on Cancer (IARC), the age-standardized incidence of prostate cancer in China is 1.6/10(5) person years (PY), with a mortality rate of 1.0/10(5) PY and mortality-to-incidence rate ratio (MR/IR) = 0.63. The MR/IR ratio of prostate cancer in China was found to be higher than the average in Asia (MR/IR = 0.57) and much higher than that in North America (MR/IR = 0.13). These data indicate that in China most prostate cancers were in the advanced stages at the time of diagnosis, and that patients had a short survival time thereafter. In 2004, Stamey et al. reported a retrospective American study of prostate cancer for the years 1983-2003. It was shown that most cases of prostate cancer detected by prostate-specific antigen (PSA) screening were in the advanced stage at the start of this 20-year period. These early follow-up data are quite similar to the results obtained from mass PSA screening of elderly men in Changchun, China. However, after the American programmes for early diagnosis and treatment of prostate cancer were accepted, tumours were diagnosed at earlier stages. On the basis of these findings, mass screening should be performed in the whole of China using serum PSA to facilitate early diagnosis and treatment of prostate cancer.

Hemoglobin on phosphonic acid terminated self-assembled monolayers at a gold electrode: immobilization, direct electrochemistry, and electrocatalysis.

Phosphonic acid (--PO(3)H(2)) terminated self-assembled monolayers (SAMs) on a gold surface were used as a functional interface to immobilize hemoglobin (Hb). In situ surface-enhanced infrared absorption spectroscopy (SEIRAS) measurements show that Hb immobilization is a sluggish process due to formation of multilayer Hb structures on the PO(3)H(2)-terminated SAMs, as revealed by ellipsometry, atomic force microscopy (AFM), and cyclic voltammetry (CV). In the multilayered Hb film, the innermost Hb molecules can directly exchange electrons with the electrode, whereas Hb beyond this layer communicates electronically with the electrode via protein-protein electron exchange. In addition, electrochemical measurements indicate that immobilization of Hb on the PO(3)H(2)-terminated SAMs is not driven by the electrostatic interaction, but likely by hydrogen-bonding interaction. The immobilized Hb molecules show excellent bioelectrocatalytic activity towards hydrogen peroxide, that is, the PO(3)H(2)-terminated SAMs are promising for construction of third-generation biosensors.

[Influence of expression of six transmembrane epithelial antigen of the prostate-1 on intracellular reactive oxygen species level and cell growth: an in vitro experiment].

OBJECTIVE: To investigate the STEAP1 gene function of the newly discovered gene six transmembrAne epithelial antigen of the prostate-1 (STEAP1). METHODS: Total RNA was obtained from human prostate cancer tissue and underwent PCR amplification. The full length of STEAP1 gene thus obtained was cloned. Mammalian expression vector pcDNA3. 1-STEAP1 was constructed and stably transfected into the human thyroid epithelial cells of the line FRTAP2320. A growth curve of the transfected cells was drawn. The intracellular reactive oxygen species (ROS) level was determined by flow cytometry (FC) with dichlorodihydrofluorescein diacetate (DCFHDA). RESULTS: The growth curve showed that the STEAP1 transfected cells grew faster than the control cells. FC showed that the fluorescence intensity of he intracellular ROS of the STEAP1 transfected cells was 42.13 +/- 1.13, significantly higher than those of the cell transfected with blank plasmid (10.02 +/- 1.42) and un-transfected cells (13.02 +/- 2.42, both P < 0.01). CONCLUSION: STEAP1 promotes the cell growth through inducing the intracellular ROS level.