[Joint administration of acupuncture, western and herbal medicines and bamboo-jar-cupping for stroke patients of wind-phlegm obstructing collateral type in acute stage].

OBJECTIVE: To observe the curative effect of joint administration of acupuncture, western and herbal medicines and bamboo-jar-cupping in the treatment of locomotor dysfunction in patients with apoplexy (acute phase) of wind-phlegm blocking meridian-collateral type in acute stroke patients, and its influence on some relevant laboratory indexes. METHODS: A total of 100 cases of acute stroke patients of wind-phlegm blocking meridian-collateral type were recruited, and equally and randomly divided into control group and treatment group according to the random number table. The patients of both groups received treatment of conventional western medicines (for anti-platelet aggregation, blood-lipid regulation, arterial plaque-stabilization, cerebral cell protection and blood pressure-lowering), Chinese herbal medicines (for promoting blood circulation to dredge the meridian-collaterals), and acupuncture of Neiguan (PC6), Chize (LU5), Zusanli (ST36), Binao (LI14) and Sanyinjiao (SP6); and in addition, the patients of the treatment group also treated by cupping with bamboo-jar (kept for 10 min). The treatment was conducted once a day for 2 weeks. After the treatment, the National Institute of Health Stroke Scale (NIHSS), Fugl-Meyer assessment (FMA), Barthel Index (BI), and traditional Chinese medicine (TCM) syndrome score were used to assess the state of neurofunction, locomotor function, daily living ability, and TCM symptoms. The contents of serum C-reactive protein, D-dimer and blood homocysteine were detected using radical immunodiffusion, immunoturbidimetry, and enzymic methods, respectively. RESULTS: After the treatment, of the 50 and 50 cases in the control and treatment groups, 5 and 6 were cured, 7 and 18 experienced marked improvement, 23 and 20 were effective, and 15 and 6 ineffective, with the effective rate being significantly higher in the treatment group (88.0%) than in the control group (70.0%, P<0.05). Self-comparison showed that the FMA and BI scores were significantly increased (P<0.01), and the NIHSS score and TCM syndrome score notably decreased in both groups ( P<0.01) in comparison with their own pre-treatment. Comparison between the two groups showed that the FMA and BI scores were obviously higher in the treatment group than in the control group (P<0.05), whereas the NIHSS score and TCM syndrome score as well as the C-reaction protein content evidently lower in the treatment group than in the control group (P<0.05, P<0.01). CONCLUSION: Joint administration of acupuncture, western and Chinese herbal medicines and cupping can promote the recovery of nerve function, improve locomotor function, activities of daily living and quality of life, and reduce inflammatory state in acute stroke patients with wind-phlegm blocking collaterals.

[Proteomic analysis of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells].

OBJECTIVE: To investigate the mechanism of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells. METHODS: In this study, the proteins extracted from paclitaxel-induced apoptotic MCF-7 cells were analyzed by 2-dimentional gel electrophoresis (2-DE), and compared with those from untreated MCF-7 cells. The differential proteins were identified by mass spectrometry. RESULTS: At 24 hour after paclitaxel (100 nmol/L) treatment, MCF-7 cells were collected and extracted the whole proteins. Seventeen up-regulated or down-regulated proteins were found by analysis of the differential proteomic 2-DE map. Six of them were identified by mass spectrometry. They were enolase 1, chloride intracellular channel 1, keratin 8, ribosomal protein S12, galectin-1 and histidine triad nucleotide binding protein, respectively. CONCLUSION: We effectively found the changed proteins in the process of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells by proteomic techniques. These up-regulated or down-regulated proteins are important molecules for our further research about the mechanism of paclitaxel-induced apoptosis.

Self-assembly of rod-coil-rod ABA-type triblock copolymers.

Self-assembled behavior of symmetric ABA rod-coil-rod triblock copolymer melts is studied by applying self-consistent-field lattice techniques in three-dimensional space. The phase diagram is constructed to understand the effects of the chain architecture on the self-assembled behavior. Four stable structures are observed for the ABA rod-coil-rod triblock, i.e., spherelike, lamellar, gyroidlike, and cylindrical structures. Different from AB rod-coil diblock and BAB coil-rod-coil triblock copolymers, the lamellar structure observed in ABA rod-coil-rod triblock copolymer melts is not stable for high volume fraction of the rod component (f(rod)=0.8), which is attributed to the intramolecular interactions between the two rod blocks of the polymer chain. When 0.3

Study of self-assembly of symmetric coil-rod-coil ABA-type triblock copolymers by self-consistent field lattice method.

The self-assembly of symmetric coil-rod-coil ABA-type triblock copolymer melts is studied by applying self-consistent field lattice techniques in a three-dimensional space. The self-assembled ordered structures differ significantly with the variation of the volume fraction of the rod component, which include lamellar, wave lamellar, gyroid, perforated lamellar, cylindrical, and spherical-like phases. To understand the physical essence of these phases and the regimes of occurrence, we construct the phase diagram, which matches qualitatively with the existing experimental results. Compared with the coil-rod AB diblock copolymer, our results revealed that the interfacial grafting density of the separating rod and coil segments shows important influence on the self-assembly behaviors of symmetric coil-rod-coil ABA triblock copolymer melts. We found that the order-disorder transition point changes from f(rod)=0.5 for AB diblock copolymers to f(rod)=0.6 for ABA triblock copolymers. Our results also show that the spherical-like and cylindrical phases occupy most of the region in the phase diagram, and the lamellar phase is found stable only at the high volume fraction of the rod.

A novel self-consistent-field lattice model for block copolymers.

We develop a self-consistent-field lattice model for block copolymers and propose a novel and general method to solve the self-consistent-field equations. The approach involves describing the polymer chains in a lattice and employing a two-stage relaxation procedure to evolve a system as rapidly as possible to a free-energy minimum. In order to test the validity of this approach, we use the method to study the microphases of rod-coil diblock copolymers. In addition to the lamellar and cylindrical morphologies, micellar, perforated lamellar, gyroid, and zigzag structures have been identified without any prior assumption of the microphase symmetry. Furthermore, this approach can also give the possible orientation of the rods in different structures.

A putative NES mediates cytoplasmic localization of Apoptin in normal cells.

Apoptin, a protein expressed by chicken anemia virus, is found predominantly in the cytoplasm in normal cells, whereas it localizes in the nucleus in transformed and malignant cells. However, the mechanisms that regulate the different subcellular localization of Apoptin in normal and tumor cells have not been fully clarified. In this work, a putative nuclear export signal (NES) in Apoptin was predicted. It was testified that the putative NES (pNES) of Apoptin was not a functional NES, but actually acted as a cytoplasmic retention signal. Deletion of the pNES led to the nuclear accumulation of Apoptin in normal cells. In addition, when a strong nuclear localization signal was introduced into Apoptin, it exclusively translocated to the nucleus in normal cells. These observations indicated that the cytoplasmic localization of Apoptin in normal cells results from the balance between cytoplasmic retention and nuclear import. On the other hand, the pNES was also proved to be necessary for Apoptin multimerization. Mutants lacking the pNES did not form obviously visible globular aggregates in normal or tumor cells.

[Construction of phage display antibody library to MCF-7 cells and screening of single-chain antibodies against breast cancer cells].

The aim of this study is to construct a phage display single-chain variable fragment (scFv) library against breast cancer cells and screen the specific antibodies against MCF-7 cells from the library. The BALB/C mice were immunized with MCF-7 cells. Total RNA of spleens was isolated. The heavy-chain (VH) and light-chain variable region genes (VL) of the antibodies were amplified by RT-PCR and joined into a single chain by overlapping PCR with a linker DNA encoding the peptide (Gly4Ser)3. The assembled scFv fragments were cloned into the phagemids(pCANTAB5E) and the recombinant phagemids were used to transform competent E. coli TG1. The transformed TG1 cells were infected by helper phage M13KO7 and the recombinant phagemids were rescued. The scFv fusion proteins were displayed on the surfaces of the recombinant phages. A phage display antibody library of repertoire of 1.2 x 10(6) clones was constructed. The specific antibodies against MCF-7 cells were enriched by 75 times after five rounds of affinity selection. Ten recombinant phages clones that exhibited specific binding to MCF-7 cells were identified. The specificity of those phage clones was analyzed by reactivity against HepG2 cells and Hela cells by ELISA. One of the selected phage clones against MCF-7cells was used to infect E. coli TOP10 to produce the soluble scFv antibodies after induction with IPTG. The strategy of construction and screening of antibody library directed against the whole tumor cells described in this report should be generally applicable to generate tumor cell-specific antibodies.