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F-actin rearrangement is an early event in burn-induced endothelial barrier dysfunction. HSP27, a target of p38 MAPK/MK2 pathway, plays an important role in actin dynamics through phosphorylation. The question of whether HSP27 participates in burn-related endothelial barrier dysfunction has not been identified yet. Here, we showed that burn serum induced a temporal appearance of central F-actin stress fibers followed by a formation of irregular dense peripheral F-actin in pulmonary endothelial monolayer, concomitant with a transient increase of HSP27 phosphorylation that conflicted with the persistent activation of p38 MAPK/MK2 unexpectedly. The appearance of F-actin stress fibers and transient increase of HSP27 phosphorylation occurred prior to the burn serum-induced endothelial hyperpermeability. Overexpressing phospho-mimicking HSP27 (HSP27(Asp)) reversed the burn serum-induced peripheral F-actin rearrangement with the augmentation of central F-actin stress fibers, and more importantly, attenuated the burn serum-induced endothelial hyperpermeability; such effects were not observed by HSP27(Ala), a non-phosphorylated mutant of HSP27. HSP27(Asp) overexpression also rendered the monolayer more resistant to barrier disruption caused by Cytochalasin D, a chemical reagent that depolymerizes F-actin specifically. Further study showed that phosphatases and sumoylation-inhibited MK2 activity contributed to the blunting of HSP27 phosphorylation during the burn serum-induced endothelial hyperpermeability. Our study identifies HSP27 phosphorylation as a protective response against burn serum-induced endothelial barrier dysfunction, and suggests that targeting HSP27 wound be a promising therapeutic strategy in ameliorating burn-induced lung edema and shock development., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Magnetic Fe3O4@chitosan nanoparticles were prepared by a simple in situ co-precipitation method and characterized by transmission electron microscope (TEM) and Fourier transform infrared spectroscopy (FTIR). The prepared Fe3O4@chitosan nanoparticles were used for covalent immobilization of lipase from Thermomyces lanuginosus by chemical conjugation after electrostatic entrapment (CCEE). The optimal immobilization conditions were obtained as follows: enzyme/support 19.8 mg/g, pH 5.0, time 4h and temperature 30 °C. Under these conditions, a high immobilization efficiency of 75% and a protein loading of 16.8 mg/g-support were obtained. Broad pH tolerance and high thermostability could be achieved by immobilization. The immobilized lipase retained 70% initial activity after ten cycles. Kinetic parameters Vmax and Km of free and immobilized lipase were determined as 5.72 mM/min, 2.26 mM/min and 21.25 mM, 28.73 mM, respectively. Ascorbyl palmitate synthesis with immobilized lipase was carried out in tert-butanol at 50 °C, and the conversion of ascorbic acid was obtained higher than 50%. These results showed that the immobilization of lipase onto magnetic chitosan nanoparticles by the method of CCEE is an efficient and simple way for preparation of stable lipase., (Copyright © 2015 Elsevier B.V. All rights reserved.)

High-frequency focused intravascular ultrasonic probes were fabricated in this study using dimple technique based on PMN-PT single crystal and lead-free KNN-KBT-Mn ceramic. The center frequency, bandwidth, and insertion loss of the PMN-PT transducer were 34 MHz, 75%, and 22.9 dB, respectively. For the lead-free probe, the center frequency, bandwidth, and insertion loss were found to be 40 MHz, 72%, and 28.8 dB, respectively. The ultrasonic images of wire phantom and vessels with good resolution were obtained to evaluate the transducer performance. The -6 dB axial and lateral resolutions of the PMN-PT probe were determined to be 58 μm and 131 μm, respectively. For the lead-free probe, the axial and lateral resolutions were found to be 44 μm and 125 μm, respectively. These results suggest that the mechanical dimpling technique has good potential in preparing focused transducers for intravascular ultrasound applications., (Copyright © 2014 Elsevier B.V. All rights reserved.)

In the traditional view, muscle atrophy and interstitial fibrosis were regarded as the basic pathological features of congenital muscular torticollis (CMT). But in the ultrastructure study, the mesenchyme-like cells, myoblasts, myofibroblasts, and fibroblasts were found in the proliferation of interstitium of CMT. To investigate the characteristics of pathological features and the mechanisms of muscle atrophy in CMT, we retrospectively reviewed the medical records of 185 CMT patients from July 2009 to July 2011 in Shenzhen Children's Hospital in China and performed pathological studies. According to age, the 185 CMT patients were divided into 4 groups. All resected surgical specimens were processed for hematoxylin and eosin staining and Masson trichromic staining. Sudan III staining was used for frozen sections, whereas immunohistochemical staining for S-100, calpain-1, ubiquitin, and 20S proteasome was carried out on 40 CMT specimens. Eight adductor muscle specimens from 8 patients with development dysplasia of the hip were taken as control group in the immunohistochemical staining. By Masson trichromic staining, the differences in the percent area of fibrous tissue in each CMT groups were significant. In Sudan III staining and immunostaining for S-100, adipocyte hyperplasia was the pathological feature of CMT. Moreover, compared with controls, most atrophic muscle fibers in CMT specimens were found to show strong immunoreactivity for calpain-1, ubiquitin, and 20S proteasome. With increasing age, fibrosis peaked at both sides and it was low in middle age group. Adipocytes increased with age. The characteristics of pathological features in CMT are changeable with age. The calpain and the ubiquitin-proteasome system may play a role in muscle atrophy of CMT. In the CMT, adipogenesis, fibrogenesis, and myogenesis may be the results of mesenchyme-like cells in SCM (sternocleidomastoid muscle). In conclusion, the present study furthermore supports maldevelopment of the fetal SCM theory for etiology of CMT.

Objective: To evaluate the efficacy and safety of Yindan Xinnaotong Soft Capsule (YXSC) on cerebral infarction (CI) reconvalescents of static blood blocking collaterals syndrome (SBBCS)., Methods: Totally 118 CI reconvalescents of SBBCS were randomly assigned to the test group (treated by YXSC) and the control group [treated by Naoxintong Capsule (NC)], 59 in each group. The therapeutic course for all was 12 weeks. Changes of National Institute of Health Stroke Scale (NIHSS), Barthel Index (BI), modified Rankin Scale (mRS), Chinese medical syndrome scores, and serum lipid indices were observed in the two groups., Results: Compared with the control group, the patient proportion of improving activities of daily life by more than or equal to 75 score was elevated (80.7% vs 62.5%; P < 0.05). Compared with before treatment in the same group, the NIHSS score decreased at post-treatment 4, 8, and 12 weeks in the two groups (P < 0.05). The patient proportion of dropped NIHSS score by more than or equal to 5 score was lowered (80.7% vs 57.14%), and the total effective rate of improving Chinese medical syndromes was superior in the test group after 12-week treatment (89.47% vs 71.43%, all P < 0.05). After 12-week treatment there was no statistical difference in the patient proportion of lowering mRS lower than or equal to 2 or blood lipids between the two groups (P > 0.05)., Conclusion: YXSC showed certain effect in improving activities of daily life, attenuating the neurological impairment, and elevating the total effective rate of improving Chinese medical syndromes in CI patients in the recovery stage.

Purpura fulminans (PF) is a life-threatening hemorrhagic condition. Because of the rarity and randomness of the disease, no improvement in treatment has been made for a long time. In this study, we assessed the serum proteome response to PF by comparing serum proteins between healthy controls and PF patient. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) approach was used after depleting 6 abundant proteins of serum. In total, 262 proteins were confidently identified with 2 unique peptides, and 38 proteins were identified significantly up- (≥ 2) or downregulated (≤ 0.5) based on spectral counting ratios (SpCPF/N). In the 38 proteins with significant abundance changes, 11 proteins were previously known to be associated with burn or sepsis response, but 27 potentially novel proteins may be specifically associated with PF process. Two differentially expressed proteins, alpha-1-antitrypsin (SERPINA1) and alpha-2 antiplasmin (SERPINF2), were validated by Western blot. This is the first study where PF patient and healthy controls are compared in a proteomic study to elucidate proteins involved in the response to PF. This study provides an initial basis for future studies of PF, and the differentially expressed proteins might provide new therapeutic targets to decrease the mortality of PF.

Problem: To investigate whether thymic stromal lymphopoietin (TSLP) regulates the apoptosis of decidual γδ T cells and to elucidate the mechanism., Method of Study: Primary human decidual γδ T cells were treated with TSLP only or TSLP combined with different signaling inhibitors (STAT3, STAT5, AKT, and ERK). The levels of signal transduction and activation of transcription 3 (STAT3) tyrosine phosphorylation and caspase3 expression were determined using Western blot analysis, and the apoptosis of decidual γδ T cells was analyzed by flow cytometry., Results: The proportions of γδ T cells in the peripheral circulation and in decidual CD3(+) cell population in women with normal pregnancy were higher than the proportions of γδ T cells in either non-pregnant control or miscarriage. Decidual γδ T cells co-expressed the TSLP receptors (TSLPR) and IL-7Rα, and the expression of TSLPR in decidual γδ T cells was higher than that in decidual CD8(+) and CD4(+) T cells. Treatment with TSLP significantly suppressed the apoptosis of decidual γδ T cells and enhanced STAT3 phosphorylation. Moreover, STAT3, and not other inhibitors, completely abrogated the anti-apoptotic effect and expression of caspase3 in decidual γδ T cells induced by recombinant human TSLP., Conclusion: These results suggest that TSLP may down-regulate caspase3 expression through activation of the STAT3 pathway, thereby suppressing the apoptosis of decidual γδ T cells., (© 2013 John Wiley & Sons Ltd.)

Ten single nucleotide polymorphisms were used for genotyping of 176 Tongshan Black-boned goats, which are Chinese indigenous goat colony for meat production. The average individual heterozygosity was 0.292. To assess the correlations between individual heterozygosity and growth in Tongshan Black-boned goat individuals, and the potential of using individual heterozygosity as an indicator of growth, the data of growth traits, including body weight, height at withers, body length, chest girth and cannon circumference, were collected. Significant correlations were observed between individual heterozygosity and body weight, height at withers, body length, heart girth, cannon circumference (P < 0.05). All the significant regression showed positive slope with R square values ranged from 0.0251 to 0.0368. These data suggests that individual heterozygosity is positively correlated with growth traits in Tongshan Black-boned goat individuals and associative overdominance may affect Tongshan Black-boned goat growth significantly. Therefore it is possible to use individual heterozygosity as an indicator of growth. Our results also provide a strong support to the overdominance hypothesis.

Tetraspanin CD9 has been implicated in various cellular and physiological processes, including cell migration. In our previous study, we found that wound repair is delayed in CD9-null mice, suggesting that CD9 is critical for cutaneous wound healing. However, many cell types, including immune cells, endothelial cells, keratinocytes and fibroblasts undergo marked changes in gene expression and phenotype, leading to cell proliferation, migration and differentiation during wound repair, whether CD9 regulates kerationcytes migration directly remains unclear. In this study, we showed that the expression of CD9 was downregulated in migrating keratinocytes during wound repair in vivo and in vitro. Recombinant adenovirus vector for CD9 silencing or overexpressing was constructed and used to infect HaCaT cells. Using cell scratch wound assay and cell migration assay, we have also demonstrated that downregulation of CD9 promoted keratinocyte migration in vitro, whereas CD9 overexpression inhibited cell migration. Moreover, CD9 inversely regulated the activity and expression of MMP-9 in keratinocytes, which was involved in CD9-regulated keratinocyte migration. Importantly, CD9 silencing-activated JNK signaling was accompanied by the upregulation of MMP-9 activity and expression. Coincidentally, we found that SP600125, a JNK pathway inhibitor, decreased the activity and expression of MMP-9 of CD9-silenced HaCaT cells. Thus, our results suggest that CD9 is downregulated in migrating keratinocytes in vivo and in vitro, and a low level of CD9 promotes keratinocyte migration in vitro, in which the regulation of MMP-9 through the JNK pathway plays an important role.

Hypoxia-induced microtubule disruption and mitochondrial permeability transition (mPT) are crucial events leading to fatal cell damage and recent studies showed that microtubules (MTs) are involved in the modulation of mitochondrial function. Dynein light chain Tctex-type 1 (DYNLT1) is thought to be associated with MTs and mitochondria. Previously we demonstrated that DYNLT1 knockdown aggravates hypoxia-induced mitochondrial permeabilization, which indicates a role of DYNLT1 in hypoxic cytoprotection. But the underlying regulatory mechanism of DYNLT1 remains illusive. Here we aimed to investigate the phosphorylation alteration of DYNLT1 at serine 82 (S82) in hypoxia (1% O2). We therefore constructed recombinant adenoviruses to generate S82E and S82A mutants, used to transfect H9c2 and HeLa cell lines. Development of hypoxia-induced mPT (MMP examining, Cyt c release and mPT pore opening assay), hypoxic energy metabolism (cellular viability and ATP quantification), and stability of MTs were examined. Our results showed that phosph-S82 (S82-P) expression was increased in early hypoxia; S82E mutation (phosphomimic) aggravated mitochondrial damage, elevated the free tubulin in cytoplasm and decreased the cellular viability; S82A mutation (dephosphomimic) seemed to diminish the hypoxia-induced injury. These data suggest that DYNLT1 phosphorylation at S82 is involved in MTs and mitochondria regulation, and their interaction and cooperation contribute to the cellular hypoxic tolerance. Thus, we provide new insights into a DYNLT1 mechanism in stabilizing MTs and mitochondria, and propose a potential therapeutic target for hypoxia cytoprotective studies.

Mitochondrial dysfunction of cancer cells includes increased aerobic glycolysis, elevated levels of ROS, decreased apoptosis, and resistance to chemotherapeutic agents. We hypothesized that the introduction of normal mitochondria into cancer cells might restore mitochondrial function and inhibit cancer cell growth, and reverse chemoresistance. First, in the present study, we tested if mitochondria of immortalized, untransformed mammary epithelial MCF-12A cells could enter into human cancer cell lines. Second, if introducing normal mitochondria into cancer cells would inhibit proliferation. And third, would the addition of normal mitochondria increase the sensitivity of human breast cancer MCF-7 cells to chemotherapy. We found that JC-1-stained mitochondria of immortalized, untransformed mammary epithelial MCF-12A cells can enter into the cancer cell lines MCF-7, MDA-MB-231, and NCI/ADR-Res, but cannot enter immortalized, untransformed MCF-12A cells. The normal mitochondria from immortalized, untransformed MCF-12A cells suppressed the proliferation of MCF-7 and NCI/ADR-Res cells in a dose-dependent pattern, but did not affect the proliferation of immortalized, untransformed MCF-12A cells. The normal mitochondria from immortalized, untransformed MCF-12A cells increased the sensitivity of human breast cancer MCF-7 cells to doxorubicin, Abraxane, and carboplatin. In conclusion, the introduction of normal mammary mitochondria into human breast cancer cells inhibits cancer cell proliferation and increases the sensitivity of the MCF-7 human breast cancer cell line to doxorubicin, Abraxane, and carboplatin. These results support the role of mitochondrial dysfunction in cancer and suggest the possible use of targeted mitochondria for cancer therapeutics.

Melatonin and its receptors have been detected in the ovary of many species, and mediate ovarian functions. The present study was designed to investigate the expression and subcellar location of melatonin receptors in bovine granulosa cells (GCs), using reverse transcription (RT) polymerase chain reaction, Western blot, and immunofluorescence analyses. Furthermore, expression level of melatonin receptors mRNA (real-time polymerase chain reaction) after treatment with various concentrations of melatonin, as well as its effects on cell apoptosis, proliferation, and steroidogenesis (by flow cytometry and RIA), were determined. In bovine GCs, melatonin receptors MT1 and MT2 were differentially located at the cell membrane, the cytoplasm, and nuclear membranes. The expression of MT1 and MT2 mRNA was regulated differently by melatonin in time- and dose-dependent manners. Exogenous melatonin suppressed cell apoptosis (P < 0.05) but not proliferation (P > 0.05). After 72 h, the apoptotic rate was significantly inhibited in all treatment groups. Meanwhile, melatonin supplementation stimulated progesterone production, but inhibited estradiol biosynthesis, in a time-dependent manner. Progesterone production was highest (P < 0.05) at 72 h. Estradiol concentrations were almost unaffected (P > 0.05) at 24 h, but were decreased (P < 0.05) at 48 h. In conclusion, exogenous melatonin acts via receptors and has important roles in regulation of development and function of bovine GCs., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Objective: To investigate the relationship of erectile dysfunction (ED) with blood vessel-, nerve- and androgen-related factors in young and middle-aged men with type 2 diabetes mellitus (T2DM) in order to provide some clinical evidence for early prevention and treatment of ED., Methods: We divided 53 male T2DM patients under 50 years into an ED group (IIEF-5 score < or = 21, n = 28) and a non-ED (NED) group (IIEF-5 score > or = 22, n = 25). We detected the levels of blood lipid, glucose, total testosterone (TT), sex hormone-binding globulin (SHBG), sulfate dehydroepiandrosterone (DHEA-S), calculated free testosterone (cFT), and examined the complications of macroangiopathy (MA), diabetic retinopathy (DR) and diabetic peripheral neuropathy (DPN), and compared the above indicators between the two groups., Results: There were no significant differences between the two groups in age, diabetes duration, body mass index, blood pressure, and blood lipid and glucose levels (P > 0.05). The incidence rate of DR was significantly higher in the ED than in the NED group (39.3% vs 4.0%, P < 0.05), but no statistically significant differences were found in the levels of TT, cFT, SHBG and DHEA-S and the incidence rates of MA and DPN between the two groups (P > 0.05)., Conclusion: The incidence of ED is closely related to DR in young and middle-aged men with T2DM. Therefore particular attention should be paid to the erectile function of T2DM patients with DR as early as possible.

Objective: To investigate mutations of anaplastic lymphoma kinase (ALK) in Chinese children with neuroblastoma (NB)., Methods: Genomic DNA was extracted from 22 cases of paraffin-embedding NB tumor tissues. Gene mutations in the exons 20-26 which were mutational hotspots of ALK were analyzed by PCR-DNA direct sequencing., Results: A novel synonymous mutation C3586T (Leu1196Leu) and a known synonymous mutation C3375A (Gly1125Gly) were found and located at exon 23 and exon 21 of ALK respectively. There were 10 cases (46%) of known synonymous mutation C3375A in 22 cases of NB. The C3375A allelic frequency was 27%. No statistically significant correlation was found between mutation C3375A and clinical parameters of NB such as age, sex, metastasis and tumor differentiation. Mutation was not found in the other 5 exons., Conclusions: A novel ALK gene synonymous mutation C3586T was identified using PCR-DNA sequencing. A known mutation C3375A in ALK was successfully identified in children, and its incidence is not influenced by the clinical features of childhood NB.

The asymmetric unit of the title compound, C(15)H(17)NO(4)·H(2)O, contains two organic mol-ecules with marginal differences between them and two water molecules. The chromine rings in both mol-ecules are essentially planar, with maximum deviations of 0.012 (2) and 0.060 (2) Å. The five-membered cyclo-pentane rings adopt envelope conformations in both mol-ecules. In the crystal, the components are linked by N-H⋯O, O-H⋯O and C-H⋯O hydrogen bonds, resulting in a three-dimensional network.

Chymosin can specifically break down the Phe105-Met106 peptide bond of milk κ-casein to form insoluble para-κ-casein, resulting in milk coagulation, a process that is used in making cheese. In this study, in order to obtain an alternative milk coagulant which is safe and efficient, and simultaneously can produce cheese with a good taste, bovine prochymosin B was chosen and constitutively expressed to a high level in Pichia pastoris. The recombinant chymosin was expressed mainly as a secretory form, and it exhibited milk-clotting activity. It was purified by ammonium sulfate fractionation, anion exchange, followed by cation exchange chromatography. A final yield of 24.2% was obtained for the purified enzyme, which appeared as a single band in SDS-PAGE having a molecular mass of approximate 36 kDa. Proteolysis assay showed that it specifically hydrolyzed κ-casein. It was stable at 25-50°C and had optimal activity at 37°C and pH 4.0. The activity of the recombinant chymosin was activated by cations such as Mn(2+), Fe(3+), Mg(2+) and Na(+), but inhibited by K(+), Co(2+), Zn(2+), Ni(2+), and to a lesser extent by Cu(2+). These results suggested that recombinant bovine chymosin is an acid milk coagulant, and it could be considered as a safe and efficient enzyme suitable for use in cheese production.

Background/aims: Nicotinamide plays a protective role in hypoxia-induced cardiomyocyte dysfunction. However, the underlying molecular mechanisms remain poorly understood. The purpose of this study was to investigate these and the effect of nicotinamide pretreatment on hypoxic cardiomyocytes., Methods: Cultured rat cardiomyocytes were pretreated with nicotinamide, subjected to hypoxia for 6 h, and then cell necrosis and apoptosis were examined. The effects of nicotinamide pretreatment on hypoxia-induced reactive oxygen species (ROS) formation, antioxidant enzyme expression, nicotinamide adenine dinucleotide (NAD(+)) and nicotinamide adenine dinucleotide phosphate (NADP(+)) levels, adenosine triphosphate (ATP) production and mitochondrial membrane potential were tested to elucidate the underlying mechanisms., Results: Based on the findings that nicotinamide treatment decreased protein expression of receptor-interacting protein (RIP; a marker for cell necrosis) and cleaved caspase-3 (CC3; a marker for cell apoptosis) in normoxic cardiomyocytes, we found that it dramatically reduced hypoxia-induced necrosis and apoptosis in cardiomyocytes. The underlying mechanisms of these effects are associated with the fact that it increased protein expression of superoxide dismutase and catalase, increased intracellular levels of NAD(+) and ATP concentration, decreased mitochondrial ROS generation and prevented the loss of mitochondrial membrane potential., Conclusion: All of these results indicate that nicotinamide pretreatment protects cardiomyocytes by improving mitochondrial stress. Our study provides a new clue for the utilization of nicotinamide in therapies for ischemic heart disease., (Copyright © 2012 S. Karger AG, Basel.)

Background: Our previous research found that structural changes of the microtubule network influence glycolysis in cardiomyocytes by regulating the hypoxia-inducible factor (HIF)-1α during the early stages of hypoxia. However, little is known about the underlying regulatory mechanism of the changes of HIF-1α caused by microtubule network alternation. The von Hippel-Lindau tumor suppressor protein (pVHL), as a ubiquitin ligase, is best understood as a negative regulator of HIF-1α., Methodology/principal Findings: In primary rat cardiomyocytes and H9c2 cardiac cells, microtubule-stabilization was achieved by pretreating with paclitaxel or transfection of microtubule-associated protein 4 (MAP4) overexpression plasmids and microtubule-depolymerization was achieved by pretreating with colchicine or transfection of MAP4 siRNA before hypoxia treatment. Recombinant adenovirus vectors for overexpressing pVHL or silencing of pVHL expression were constructed and transfected in primary rat cardiomyocytes and H9c2 cells. With different microtubule-stabilizing and -depolymerizing treaments, we demonstrated that the protein levels of HIF-1α were down-regulated through overexpression of pVHL and were up-regulated through knockdown of pVHL in hypoxic cardiomyocytes. Importantly, microtubular structure breakdown activated p38/MAPK pathway, accompanied with the upregulation of pVHL. In coincidence, we found that SB203580, a p38/MAPK inhibitor decreased pVHL while MKK6 (Glu) overexpression increased pVHL in the microtubule network altered-hypoxic cardiomyocytes and H9c2 cells., Conclusions/significance: This study suggests that pVHL plays an important role in the regulation of HIF-1α caused by the changes of microtubular structure and the p38/MAPK pathway participates in the process of pVHL change following microtubule network alteration in hypoxic cardiomyocytes.

Interleukin-6 (IL-6) plays an important role in the neoplastic process through its action on cancer cell adhesion, motility, proliferation, tumor-specific antigen expression, and thrombopoiesis. IL-6 exerts its activity by binding to a high affinity receptor complex consisting of two membrane glycoproteins: the 80 kDa IL-6 a-receptor subunit (IL-6R) and the 130 kDa signal-transducing protein (GP130). In the present study, MCF-7 breast cancer cells were cultured with human IL-6 and IL-6 soluble receptor (sIL-6R). MCF-7 cells were also treated with either antibodies specific to human IL-6 and IL-6R, or synthetic antisense oligodeoxynucleotides (ODNs) targeted to IL-6 and IL-6R genes. Cell growth was measured, and it was found that human IL-6 and sIL-6R did not significantly increase the proliferation of MCF-7 cells. When IL-6 produced by the MCF-7 cells was bound by rabbit anti-human IL-6 antibody, there was a significant dose-dependent inhibition of cell proliferation. IL-6 and IL-6R antisense ODNs caused a marked and specific decrease in IL-6 and IL-6R mRNA and proteins, respectively. Both IL-6 and IL-6R antisense ODNs significantly inhibited the proliferation of MCF-7 cells, but the inhibitory effect of IL-6R antisense ODN was greater than that of IL-6 antisense ODN (IC₅₀: IL-6R: 1 μM; IL-6: 5 μM, 72-hour incubation). Addition of exogenous IL-6 partially reversed the growth inhibition caused by IL-6 antisense ODN but not the growth inhibition caused by IL-6R antisense ODN. In conclusion, IL-6 plays an important role in maintaining the growth of MCF-7 breast cancer cells. These results suggest careful modulation of IL-6 and IL-6R expression of cells as a potential approach for breast cancer therapy.

Human leukocyte antigen G (HLA-G) is an immunotolerant nonclassical major histocompatibility complex Class Ib molecule. It is expressed by trophoblastic placental cells during pregnancy to protect the fetus from maternal alloreactivity. HLA-G is overexpressed in tumors and involved in cancer immune evasion. Reverse transcription-polymerase chain reaction and immunohistochemistry (IHC) were used to examine HLA-G expression in normal mammary and breast cancer cell lines and normal and human breast cancer tissues. Reverse transcription-polymerase chain reaction confirmed that normal epithelial MCF-12A cells had no HLA-G mRNA expression, whereas cancer cell lines MCF-7, T47D, and MDA-MB-231 and NCI/Adr-Res had various levels of HLA-G mRNA expression. Twelve (12) normal and 38 breast cancer tissues were examined by IHC. Fifty-eight (58) percent (22/38) of cancers had medium to strong staining to HLA-G, whereas only 8% (1/12) of normal breast tissues had medium to strong staining, and the difference was significant (p < 0.05). HLA-G staining was found in the membranes and cytoplasm of cancer cells. In conclusion, breast cancer cells overexpress HLA-G mRNA and protein, and this probably contributes to immune evasion.

Respectively using pressed method and diffuse reflectance method, the infrared absorption spectrua of sorts of kaolin in various areas were obtained. The relationship between the structure characteristics of kaolin and its absorption peaks was analyzed. The results showed that compared with pressed method, the use of diffuse reflectance Fourier transform infrared spectroscopy, combined with the K-M function, results in corrected infrared spectra with high sensitivity and is more accurate and easier to analyze. Furthermore, based on a high frequency wave of 3 700-3 600 cm(-1) sections of kaolinite-OH characteristic absorption peaks, the crystallinity of all kinds of kaolnite can be known quickly. The result is consistent with the Hinckley index surveyed by X-ray diffraction technique.

The five-membered ring of the title compound, C(11)H(9)ClF(3)N(3), is almost planar (r.m.s. deviation = 0.002 Å) and the phenyl-ene ring is aligned at 44.8 (1)°. The N atom of the amino substituent shows a pyramidal geometry and is a hydrogen-bond donor to a Cl atom and to a ring N atom, which together generate a layer motif.

Lactoferrin (LF) is a member of the transferrin family that is abundantly expressed and secreted by glandular epithelial cells. The biological functions of LF involve in iron homeostasis regulation of the body and antibacterial activity. Previous studies demonstrated that it had a high cationic N-terminal domain that could interact with glycosaminoglycans, lipopolysaccharides and the bacterial virulence protein. Two anti-microbial peptides, lactoferricin (LFcin) and lactoferrampin (LFampin), were also isolated and identified in N-terminal of LF. Although the antibacterial mechanism was carefully studied, little was known about the molecular evolution of LF. In this study, we estimated the nonsynonymous-to-synonymous substitution ratios ( ) per site using maximum likelihood method to analyze the LF evolution. The results of omega > 1 and five identified positive selection sites of amino acid suggested that the evolution of LF gene was characterized by positive selection. Further study found that the positive selection sites were either located in the LF-bacteria binding region or the peptides of LFcin and LFampin, indicating that the selection pressure was related to LF-bacteria interaction. The identification of these sites may contribute to the mechanism of bacteria-LF interaction.

Since malignant cells often have a high demand for iron, we hypothesize that breast cancer cells may alter the expression of iron transporter genes including iron importers [transferrin receptor (TFRC) and solute carrier family 11 (proton-coupled divalent metal ion transporters), member 2 (SLC11A2)] and the iron exporter SLC40A1 (ferroportin), and additionally that the growth of breast cancer can be inhibited by manipulating iron transporter gene expression. To test our hypothesis, reverse transcription polymerase chain reaction (RT-PCR) was used to determine mRNA expression of iron transporter genes in normal human mammary epithelial MCF-12A cells and human breast cancer MCF-7 cells. Antisense oligonucleotides were employed to suppress the expression of TFRC gene in the 4T1 mammary adenocarcinoma in both cell culture and a mouse tumor model. We found the following: i) the MCF-7 cells have higher expression of TFRC and SLC11A2 compared with MCF-12A epithelia; ii) SLC40A1 was only expressed in MCF-12A epithelia but not in MCF-7 cells; iii) iron increased mRNA levels of the SLC11A2 gene in both MCF-12A and MCF-7 cells; iv) TFRC antisense oligonucleotides reduced TFRC mRNA levels and intracellular total iron, and inhibited the proliferation of the 4T1 cells in cell culture; v) TFRC antisense oligonucleotide inhibited tumor growth and lung metastases in the 4T1 mammary adenocarcinoma mouse model. In conclusion, breast cancer cells up-regulate the expression of iron importer genes and down-regulate the expression of iron exporter SLC40A1 to satisfy their increased demand for iron. Suppression of transferrin receptor by antisense results in inhibition of tumor growth and lung metastasis in the 4T1 mammary adenocarcinoma mouse model.

Objective: To observe the effect of acupoint catgut embedding on height of adolescents with spleen qi deficiency., Methods: Three hundred voluntary adolescents with spleen qi deficiency syndrome who want to increase height were stratified according to sex first, and then randomly divided into an observation group, a control group and a placebo group, 100 cases in each group. The observation group was treated with acupoint catgut embedding at Zusanli (ST 36), Pishu (BL 20), Weishu (BL 21) etc. The control group was treated with oral administration of Lysine Dicalcium Phosphate particles, and the placebo group was treated with oral administration of flour capsule. The height was measured and the therapeutic effects were detected after treatment of 3 months and 6 months respectively., Results: The total effective rate of 95.0% (95/100) in the observation group was superior to that of 49.0% (49/100) in the control group and 0 (0/100) in the placebo group (both P < 0.05). The height increased more obviously in the observation group than that of the control group and the placebo group, there were significant differences among three groups (all P < 0.01)., Conclusion: Acupoint catgut embedding can increase the height and improve the symptoms of the adolescents with slow growth caused by spleen qi deficiency.

Objective: To evaluate the effects of mechanical ventilation on pulmonary function during short duration of general anesthesia with tracheal intubation, and assess the safety of controlled spontaneous respiration during general anesthesia., Methods: Fifty-three adult patients (aged 18-55 years, ASA physical status I-II) scheduled for elective unilateral tympanoplasty were randomly assigned into mechanical ventilation group (group M, n=28) and spontaneous respiration group (group S, n=25). Anesthesia induction was performed in group M with intravenous propofol (2 mg/kg), fentanyl (3 microg0.05). The pH and SpO(2) [ (97.9-/+1.00)% at the lowest] and PaO(2) in group S were significantly lower and the PaCO(2) was higher than those in group M (P<0.05). In group S, the pH values were 7.274-/+0.025 and 7.331-/+0.039, PaCO(2) values were 60-/+6 and 53-/+5 mmHg, and PETCO(2) values were 53-/+ 6 and 48-/+7 mmHg, and the PaO(2) values were 143-/+37 and 165-/+49 mmHg immediately and 150 min after the intubation, respectively. These values were considered safe under the concept of permissive hypercapnia. No significant differences were found in the P(A-a)DO(2), RI, VD/VT and TFC between or within the two groups (P>0.05), nor were moving, bucking, swallowing and awareness recorded during the surgical procedures., Conclusion: In essentially normal lungs, short-term mechanical ventilation during general anesthesia with tracheal intubation does not damage the lung functions, and spontaneous respiration can offer sufficient oxygen supply without causing harmful carbon dioxide retention.

1. The objective of this study was to determine hens' sperm storage potential. 2. Efficient duration (De, number of days between insemination and the first clear egg), maximum duration (Dm1, number of days from the day after insemination to the last fertile egg before two consecutive infertile eggs) and maximum duration (Dm2, number of days from the day after insemination to the last fertile egg) of fertility, fertile egg number, egg production, laying rate and fertility during the 24 d following the latter of two inseminations (with 1 x 10(8) spermatozoa) on two consecutive days were measured in a total of 150 dual-purpose hens at 30 weeks of age. 3. De, Dm1 and Dm2 were 12.06, 14.44 and 16.17 d, respectively, and the three definitions of fertility duration (DF) differed greatly. 4. Significant correlation coefficients between De and Dm1, Dm1 and Dm2, and De and Dm2 were 0.51, 0.57 and 0.23, respectively. 5. We suggest that De and fertile egg number should be used to assess the ability of storing spermatozoa in female fowl.

A general and versatile method for the divergent and diastereoselective synthesis of polyhydroxylated indolizidines has been established. The annulation reactions of a readily available enantiopure dihydroxylated cyclic secondary enamine with alpha,beta-unsaturated carboxylates including methyl acrylate, methyl crotonate, methyl 2-hexenoate, allenoate, and dimethyl acetylenedicarboxylate and with malonyl chloride produced hexahydro- or tetrahydro-5-indolizinone-8-carboxylates in high yields. The resulting 5-indolizinone derivatives were converted into diverse polyhydroxylated indolizidines in good yields through practical hydrogenation and reduction reactions.

In this article, a multilayer piezoelectric transformer based on lead-free Mn-doped 0.94(Bi(12)Na(12))TiO(3)-0.06BaTiO(3) ceramics is presented. This piezoelectric transformer, with a multilayered construction in the thickness direction, is 8.3 mm long, 8.3 mm wide, and 2.3 mm thick. It operates in the second thickness extensional vibration mode. For a temperature rise of 20 degrees C, the transformer has an output power of >0.3 W. With a matching load resistance of 10 Omega, its maximum efficiency approaches 81.5%, and the maximum voltage gain is 0.14. It has potential to be used in low voltage power supply units such as low power adapter and other electronic circuits.

A total of 139 hybrid finishing pigs from Large White x Meishan were slaughtered at about 88kg body weight. Fourteen meat quality traits and 8 carcass traits were assayed for each pig. FUT1 gene was scored by PCR-RFLP. The values of meat pH and meat color for AA genotype pig were higher than those of AG genotype pig (P<0.05). Water holding capacity (WHC) of AA genotype pig was higher than that of AG genotype pig(91.02% VS 86.70%, P<0.05). Backfat at 6-7 vertebra (BV) and backfat at vertebra-lumbar (BVL) of AA genotype pig were lower than those of AG genotype pig 4.26 mm and 3.96 mm respectively (P<0.05). Meat factor of AA genotype pig was higher than that of AG genotype pig 3.31% (53.46% VS 50.15%, P<0.05). These results indicated that FUT1 gene had good genetic effects on pig meat quality and carcass traits.

We report the anesthetic management of a case of separation of craniopagus twins with unbalanced cross circulation and one twin with renal dysfunction. After intravenous induction, anesthesia was maintained with isoflurane inhalation and propofol infusion. Twin A survived but Twin B died after the surgery. The anesthetic problems during the operation are discussed.

382 yak cows were examined for milk yield, fat, protein and lactose contents. Six polymorphic loci, alphas1-CN, kappa-CN, beta-CN, beta-Lg, alpha-La and MUC-1, were scored by PAGE electrophoresis for each individual. The values of milk yield, fat, protein and lactose content were 247.13 kg, 5.81%, 5.18% and 4.93%, respectively. Based on the 6 polymorphism loci, the average heterozygosity of the yak population was 0.1794. Calculated by the marker-based method, heritability estimates for milk yield, fat, protein and lactose contents were 0.353 +/- 0.093, 0.316 +/- 0.101, 0.415 +/- 0.098 and 0.481 +/- 0.035, respectively. The relatively high or medium heritability of these traits indicate that it is feasible to rely directly on them in breeding for the improvement in a relatively short period. The significant linear regression between heterozygosity and fat percentage with a positive slope (R = 0.0420) indicated that inbreeding affected milk fat content in this population.

Genotypes of 24 microsatellite loci were assayed in total of 139 F2 finishing pigs from resource population(Large White x Meishan) in the present study. There were not significant correlation in sex, family and microsatellite loci effects on birth weight (BW), average daily gain (ADG), adjusted weight of 180 d (LWT180), relative growth rate (RGR) and kleiber ratio (KR). Individual gene heterozygosity (Hi) was estimated by using 24 microsatellites. The 139 individuals were classified into five clusters based on Hi by stepwise 0.05 and the traits mentioned above were compared between Hi levels. Birth weight decreased significantly from gene heterozygosity level 1(0.5101 Hi and 1.5185 kg) to level 2(0.5796 Hi and 1.3063 kg, P < 0.05), the difference was 0.2123 kg equal to 14.66% of population average birth weight (1.4479 kg). And then the birth weight increased significantly with the gene heterozygosity increasing (P < 0.05). There were nonlinear relationships between LWT180, ADG, RGR, KR and individual gene heterozygosity, and mostly like the pattern of sinusoidal curve. The peaks were located at about heterozygosity level 2(averagely Hi 0.5796). ADG, RGR and KR at heterozygosity level 2 were significantly higher than level 4(28.7 g/d, 0.0375 and 0.0003 respectively, P < 0.05). The differences were equal 7.38%, 3.99% and 2.20% to respective average population values.

Objective: To investigate serum levels of endogenous oxidation agents, anti-oxidation agents and clinical significance in the patients with cerebral vascular disease (CVD)., Methods: Using biochemical methods, the levels of serum nitric oxide (NO), nitric oxide synthase (NOS), malondialdehyde (MDA) and anti-oxidants vitamin E (VitE), vitamin C (VitC), superoxide dismutase (SOD) in 49 patients with cerebral hemorrhage (CH), 65 patients with cerebral infarction(CI) and 35 patients with other nervous system diseases and 34 healthy controls were determined., Results: In CH and CI groups, the levels of serum NO and MDA and the activity of serum NOS were significantly higher than that of the two other groups (P<0.05 or P<0.01). On the other hand, the patients with CH and CI had lower VitE, VitC levels and SOD activity than that of the two control groups (P<0.05 or P<0.01)., Conclusion: These findings suggest NO and NOS plays an important role in pathogenesis of cerebral damage after CH and CI. Determination of the concentrations of NO, VitE, VitC, MDA level, and NOS and SOD activity in serum can also help judge the seriousness and the course of the disease.

An intercross between Meishan and Large White pig population was used to map quantitative trait loci(QTL) for protein percentage in muscle. In the present study, all 135 F2 animals, their parents and their grandparents were genotyped for 24 microsatellites, which were in 1, 2 and 6 chromosome. The marker genotypes were used to calculate the additive and dominant coefficients at fixed position in the genome of each F2 animal, and the protein percentage were regressed on to the coefficients in intervals of 1 cM. Three significant QTLs effects were found for protein percentage in muscle. They were located between markers Sw1332 and Sw2130 at chromosome 1, between Sw2516 and Sw1201 at chromosome 2 and between Sw322 and Sw607 at chromosome 6(P < 0.05), and the explained residual variance were 0.91%, 12.76% and 4.60%, respectively. The results show that QTLs, which affect protein percentage, may be located in different chromosomes.

A high performance liquid chromatographic method was established for the determination of cinnabar in Zijinhong capsules using diethyldithiocarbamate (DEDTC) chelate. A Waters X-Terra C18 column was used with a mixture of methanol-0.01 mol/L Na2HPO4 (pH 7.5) (containing 0.01% DEDTC) (73:27, V/V) as the mobile phase and a detector set at 270 nm. The calibration curve was linear (r = 0. 999 6) in the range of 10.1 mg/L - 100.9 mg/L of Hg2+. The recoveries were 97.0% - 100.8% with RSDs of 1.8% - 2.3%.

It is known that the interruption of normal iron metabolism with chelators of iron, toxic metals, toxic metals bound to transferrin, or anti-transferrin receptor antibodies leads to significant inhibition of tumor cell growth in cell culture systems and animal models. In the present study, we found that iron depletion was produced by the iron chelator deferoxamine mesylate, the free toxic metals gallium or indium, and the toxic metals gallium or indium bound to transferrin in the MCF-7 human breast cancer cell line, and this induced the condensation and fragmentation of chromatin, and the formation of DNA fragments characteristic of apoptosis. The induction of apoptosis was quantitated with acridine orange and ethidium bromide staining of apoptotic cells, separation of fragmented DNA from radiolabeled cells, and in situ terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assays. The apoptosis, caused by deferoxamine mesylate, and gallium or indium bound to transferrin in the MCF-7 cells, can be completely inhibited by excess ferric chloride or equimolar iron-loaded transferrin. Gallium-transferrin and indium-transferrin complexes induced more apoptosis than their respective salts in the MCF-7 cells. Deferoxamine mesylate induced a small increase in the endogenous expression of both the bcl-2 and bax genes in the MCF-7 cells and this can be prevented by ferric chloride. In the 13762NF rat mammary adenocarcinoma model, in situ TUNEL assays showed that the iron-deficiency following a low iron diet or intravenous injection of deferoxamine mesylate produced 5.32 +/- 3.90% and 6.46 +/- 3.58% of apoptotic cells, respectively, compared to 2.01 +/- 1.20% of apoptotic cells in the control rats maintained on a normal diet (p < 0.05 and p < 0.01, respectively, Student's t-test). This is the first report of iron depletion caused by a low iron diet or deferoxamine mesylate treatment inducing apoptosis in rats bearing the 13762NF marnmary adenocarcinoma.

NMDA glutamate receptor antagonists are used in clinical anesthesia, and are being developed as therapeutic agents for preventing neurodegeneration in stroke, epilepsy, and brain trauma. However, the ability of these agents to produce neurotoxicity in adult rats and psychosis in adult humans compromises their clinical usefulness. In addition, an NMDA receptor hypofunction (NRHypo) state might play a role in neurodegenerative and psychotic disorders, like Alzheimer's disease and schizophrenia. Thus, understanding the mechanism underlying NRHypo-induced neurotoxicity and psychosis could have significant clinically relevant benefits. NRHypo neurotoxicity can be prevented by several classes of agents (e.g. antimuscarinics, non-NMDA glutamate antagonists, and alpha(2) adrenergic agonists) suggesting that the mechanism of neurotoxicity is complex. In the present study a series of experiments was undertaken to more definitively define the receptors and complex neural circuitry underlying NRHypo neurotoxicity. Injection of either the muscarinic antagonist scopolamine or the non-NMDA antagonist NBQX directly into the cortex prevented NRHypo neurotoxicity. Clonidine, an alpha(2) adrenergic agonist, protected against the neurotoxicity when injected into the basal forebrain. The combined injection of muscarinic and non-NMDA Glu agonists reproduced the neurotoxic reaction. Based on these and other results, we conclude that the mechanism is indirect, and involves a complex network disturbance, whereby blockade of NMDA receptors on inhibitory neurons in multiple subcortical brain regions, disinhibits glutamatergic and cholinergic projections to the cerebral cortex. Simultaneous excitotoxic stimulation of muscarinic (m(3)) and glutamate (AMPA/kainate) receptors on cerebrocortical neurons appears to be the proximal mechanism by which the neurotoxic and psychotomimetic effects of NRHypo are mediated.

N-methyl-D-aspartate (NMDA) glutamate receptor antagonists are used in clinical anesthesia and are being developed as therapeutic agents for preventing neurodegeneration in stroke, epilepsy, and brain trauma. However, the ability of these agents to produce neurotoxicity in adult rats and psychosis in adult humans compromises their clinical usefulness. In addition, an NMDA receptor hypofunction (NRHypo) state might play a role in neurodegenerative and psychotic disorders, like Alzheimer's disease, bipolar disorder and schizophrenia. Thus, developing pharmacological means of preventing these NRHypo-induced effects could have significant clinically relevant benefits. NRHypo neurotoxicity appears to be mediated by a complex disinhibition mechanism that results in the excessive stimulation of certain vulnerable neurons. Here we report our findings that five agents (phenytoin, carbamazepine, valproic acid, lamotrigine, and riluzole), thought to possess anticonvulsant activity because they inhibit voltage-gated sodium channels, prevent NRHypo neurotoxicity. The ability of tetrodotoxin, a highly selective inhibitor of voltage-gated sodium channels, to prevent the same neurotoxicity suggests that inhibition of this ion channel is the likely mechanism of action of these five agents. We also found that three other anticonvulsants (felbamate, gabapentin and ethosuximide), whose mechanism is less clear, also prevent NRHypo neurotoxicity, suggesting that inhibition of voltage-gated sodium channels is not the only mechanism via which anticonvulsants can act to prevent NRHypo neurotoxicity. Several of these agents have been found to be of clinical use in bipolar disorder. It would be of interest to determine whether these agents might have therapeutic benefits for conditions in which a NRHypo state may exist.

Transferrin receptor expression is controlled by the amount of iron required by the cell to maintain its metabolism and therefore tumor cells in a highly proliferative state have a high density of transferrin receptors. In this study, phosphorothioated antisense TfR oligonucleotides (TfR-ODna) targeted to the sequences of TfR mRNA including the AUG initiation codon and the control sense chain (TfR-ODns) were synthesized. The rate of cellular DNA synthesis was determined by [3H]-thymidine incorporation. Administering TfR-ODna to three morphologically distinct breast cancer cell lines (MCF-7, T47D, and MDA-MB-231) and a normal breast cell line (MCF-12A) caused specific inhibition of tumor cell growth. The IC50 (50% inhibition of DNA synthesis) of the TfR-ODna for the MCF-7, T47D and MDA-MB-231 cells were 0.5, 0.5, and 1.0 microM, respectively, whereas the MCF-12A normal breast cells were about 30 times (IC50 of 30 microM) less sensitive to TfR-ODna than the breast cancer cells. The cytotoxicity of the antisense TfR-ODna was 10 to 60 times greater than that of the sense chain (TfR-ODns). TfR mRNA and protein synthesis were demonstrated by RT-PCR and immunohistochemical staining, respectively. Approximately 50% inhibition of the expression of TfR mRNA was observed when breast cancer cells were treated with 1 microM antisense TfR ODNa for 72 hrs but 1 microM antisense only caused 14% inhibition in normal breast cells. The decreased cytotoxicity and inhibition of TfR gene expression when the tumor cells were treated with the same concentration (1 microM) of TfR-ODns demonstrated the specificity of the TfR-ODna for blocking the target TfR gene. The combined cytotoxicities to human breast tumor MCF-7 cells of the antisense TfR-ODna and the iron chelator deferoxamine (DFO) or the ribonucleotide reductase inhibitor hydroxyurea were observed in this study. IC50s (50% inhibition of DNA synthesis) for DFO and hydroxyurea individually were 0.3 microM and 250 microM, respectively. The CalcuSyn program was used to determine the combined effects among the agents and synergism (Combined Indexes (CI) < 1) were found with the following two combinations: TfR-ODna (0.007 microM to 0.15 microM) with DFO (0.15 microM to 5 microM) and TfR-ODna (0.007 microM to 0.15 microM) with hydroxyurea (50 microM to 800 microM). However, inhibition by TfR-ODns was not synergistic with either DFO or hydroxyurea. The synergistic effects on inhibition of DNA synthesis between TfR-ODna and DFO or hydroxyurea suggest that inhibition of breast cancer cell growth by TfR-ODna is produced by depletion of iron pools that are required for DNA synthesis in tumor cells. The fact that TfR-ODna specifically decreases cell viability and proliferation, and reduces TfR mRNA and protein expression in human breast carcinoma cells without affecting normal breast cells, suggests that the antisense oligonucleotide to the transferrin receptor may be a novel therapeutic approach in breast cancer.

In breast cancer there is often overexpression of the breast cancer antigen CA15-3, the carcinoembryonic antigen (CEA) and the ovarian cancer antigen CA125, which makes them potential target antigens for immunotherapy. In this study, we used a multi-antigen vaccine, which included the following antigens: autologous breast cancer cells (AUTOC), allogeneic breast cancer MCF-7 cells (ALLOC), and the tumor associated antigens CA15-3, CEA and CA125, plus low doses of granulocyte/macrophage-colony-stimulating factor (GM-CSF) and interleukin 2 (IL-2). Forty-two breast cancer patients received weekly subcutaneous vaccination at the 1st, 2nd, 3rd, 7th, 11th and 15th weeks. Their lymphocyte proliferative responses to AUTOC, ALLOC, CA15-3, CEA and CA125 were tested in lymphocyte blastogenesis assays (LBA) before and after vaccination. The disease stage and serum CA15-3, CEA and CA125 concentrations were also determined pre- and post-vaccination. We found that the vaccine was safe, and the only major side effects were swelling at the site of injection, muscle pain, and weakness or fatigue. The vaccine induced a significant increase in post-vaccination lymphocyte proliferative responses to AUTOC, CA15-3, CEA and CA125 but not ALLOC, compared to pre-vaccination (p < 0.05, p < 0.01, p < 0.05, p < 0.01 and p > 0.05, respectively, a paired t Test). Computed tomography (CT), ultrasound or bone scan showed evidence of disease improvement in 2 (12%) patients after vaccination. Hepatic metastases were reduced in size and number and some actually disappeared one patient. Metastatic disease in the L5 vertebra and the skull decreased in size and some osteolytic sites completely healed in a second patient. In addition, 7 patients (44%) had stable disease and 7 patients (44%) had disease progression. We did not find vaccination significantly reduced serum tumor markers CA15-3, CEA and CA125 of these breast cancer patients. These results suggest that the vaccine mixture of autologous and allogeneic breast cancer cells and tumor associated antigens plus GM-CSF and IL-2 can be administered safely to breast cancer patients and there is evidence for improved immunity and clinical efficacy.

Elevated serum IL-6 concentrations have been associated with poor prognosis in a variety of cancers, and decreases in serum IL-6 concentrations have been reported after chemotherapy. We have demonstrated that serum IL-6 concentrations are elevated in breast cancer patients [normal women 0.7 +/- 2.5 pg/ml (n=36), breast cancer patients 38.3 +/- 138.7 pg/ml (n = 111)]. After vaccination of breast cancer patients with a combination of tumour-associated antigens and biological adjuvants (IL-2 and GM-CSF), the concentration of IL-6 decreased significantly (P<0.05) to 8.1 +/- 14.6 pg/ml (n=85). Other studies have shown that oestrogen suppresses IL-6 production in oestrogen receptor positive breast cancer cells. We have demonstrated that the decrease in IL-6 associated with vaccination is related to the oestrogen receptor status of the tumours from breast cancer patients, as a decrease in IL-6 from 124.0 +/- 267.5 pg/ml (n=26) to 6.2 +/- 11.0 pg/ml (n=34) only occurs in patients with oestrogen receptor negative tumours. The IL-6 concentration in breast cancer patients with oestrogen receptor positive tumours remained unchanged (9.5 pg/ml before vaccination, and 9.3 pg/ml after vaccination). These results suggest that postmenopausal women with oestrogen receptor negative breast cancers, who do not respond well to either hormonal therapy with tamoxifen or adjuvant chemotherapy, may have a significant response to vaccination with autologous tumour-associated antigens.

Objective: The purpose of this study was to study the effect of splenopneumopexy for patients with portal hypertension in children., Methods: From March 1993 to April 1998, splenopneumopexy was performed on six children with portal hypertension. Doppler ultrasound and radionuclide were used to demonstrate the portopulmonary shunt after operation., Results: The bleeding from the esophageal varices was controlled and the esophageal varices were eliminated gradually. The symptoms pertaining to hypertension were disappeared. The patency of the shunt was maintained without the formation of thrombosis. No pulmonary complication was observed., Conclusion: The results indicated that splenopneumopexy was a safe and effective procedure for patients with portal hypertension in children.

Levels of expression of the nm23 gene inversely correlated with metastatic potential in several rodent tumor model systems and human breast carcinoma. In the present study, we examined nm23 mRNA levels in two murine ascites hepatoma models (H22-16A3-F and H22-A2-P) with different metastatic potentials. Metastatic H22-16A3-F (80% metastatic rate) and non-metastatic H22-A2-P clones were both derived from murine ascites hepatoma (H22). We found that a 0.8-kb nm23 transcript was expressed in both cell clones. The nm23 gene was expressed at a higher level in non-metastatic H22-A2-P: approximately 8.6-fold higher than in metastatic H22-16A3-F. The present data suggest that the expression of nm23 mRNA might be associated with metastasis of murine ascites hepatoma (H22), though heterogeneity of nm23 steady-state expression levels among the H22 clones remains to be investigated.

Nm23 is a kind of an effective tumor metastasis suppressor gene which included two types in human: nm23-H1 and nm23-H2. Amino acid identity between nm23-H1 and nm23-H2 was 88%. In this study, we used a pair of primers to flank the part of coding sequence of nm23. The 5'-translated sequence was amplified by PCR from human normal liver genomic DNA. A 375bp clone was characterized to designate pnm 23-H3b. The nm23-H3b nucleotide sequence between 40bp and 70bp was different from nm23-H1 and nm23-H2, and other sequences had 86% and 90% identical to nm23-H1 and nm23-H2, respectively. Southern blot containing Bg1II-digested human liver genomic DNA hybridized to the entire nm23-H3b DNA and showed three bands at 10.5, 7.9 and 4.0 kb. These data demonstrate that nm23-H3b is a new type of gene, which has high homology with human nm23-H1 and nm23-H2. Nm23 is possibly considered a family of closely related genes.

A new approach to perfuse arterial blood through venous channels for revascularization of severely ischemic limbs is reported. The procedure studied and used consists of creating an arteriovenous fistula between the normal arterial trunk proximal to the occlusion and the deep venous trunk of the diseased limb, constricting the venous trunk proximal to the anastomosis to one third of its lumen diameter, ligating the communicating and small tributary veins distal to the constriction in the operative field. The results of the experimental and clinical studies have shown that the treated ischemic limb was quickly revascularized without undesirable influence on cardiac function. This new approach has been used in the treatment of severe ischemia involving total 212 limbs in 156 patients, and the results appeared more satisfactory than those treated with staged arteriovenous reversal.