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The purpose of this study is to find out whether electro-acupuncture (EA) at acupoint LI.10 (Shou San Li), which has not been considered as an immune-related point, has a similar immune effect as LI.11 (Qu Chi). We recruited 24 male medical students for this study. They were randomized into 3 groups: control group (without acupuncture during the study period, n=7), LI.11 group (EA at LI.11 acupoint, n=9) and LI.10 group (EA at LI.10 acupoint, n=8). At the first day (Day 1), blood was taken from all subjects. But EA was applied to the LI.11 and LI.10 group immediately after blood drawing. On the second day (Day 2), EA was applied to the LI.11 and LI.10 group right before blood sample was taken. At the fourteenth day (Day 14), blood sample was taken for all subjects. The parameters of EA were 4 sec alternating intervals of 2 and 15 HZ (dense and disperse) and 1-4 mA in intensity for 20 min. The waveform of electrical impulse was bi-directional and pulse duration of 50 microsec. Blood analysis included flow cytometry for lymphocyte subpopulations (CD4 and CD8 positive cell percentages) and ELISA for cytokines [soluble interleukin-2 receptor (sIL-2R), interferon (INF)-gamma, IL-4, IL-6, and IL-10]. We treated the result of Day 1 as a baseline value. After EA at acupoint LI.11, CD4 positive cell percentage at Day 14 showed significant decrease compared to the baseline value (p = 0.011). CD8 positive cell percentage also decreased at Day 2 (p = 0.038) and Day 14 (p= 0.015). In the LI.10 group, CD4 and CD8 levels at Day 14 were decreased significantly (CD4, p = 0.012; CD8, p = 0.017). Results of all cytokines concentrations of the LI.11 group did not change during the study period. But the serum level of sIL-2R of the LI.10 group at Day 14 was higher than the baseline value (p = 0.018). The serum level of IL-6 at Day 14 was lower than the baseline value (p = 0.028). The serum level of IL-4 at Day 2 was lower than the baseline value (p = 0.025). Our results showed that LI.11 and LI.10 acupoints cause a similar decrease of CD4 and CD8 populations. Increase of serum sIL-2R and decrease of serum IL-4 and IL-6 were observed when EA was applied at LI.10 acupoint, but not at LI.11 acupoint. In this study, we can understand that EA may modulate the peripheral blood lymphocyte subpopulations and serum cytokine levels of the immune system.

Objective: To investigate the expression of human coagulation factor IX (hFIX) gene in human umbilical cord blood (CB) CD34+ cells which was transfected with recombinant adeno-associated virus 2 (rAAV-2)., Method: The CD34+ cells were transfected with rAAV-2/hFIX and cultured for 21 days for inducing differentiation into myeloid, erythroid and megakaryocytes, respectively. The expression of hFIX was studied at mRNA, protein and biological activity levels. The cytotoxicity of rAAV-2 to CD34+ cells was evaluated by cell proliferation, cell vitality, CD antigen expressions and CFU yields., Results: The hFIX mRNA was expressed in the cultured cells which was verified by RT-PCR and DNA sequencing. An elevated level of hFIX expression and biological activities were detected with a maximum amount of 14.10 ng/10(6) cell x 24 h. During the period of 21 day culture, the cell vitality, cell proliferation, CD antigen expression and CFU yields between the transfected and un-transfected groups had no difference(P > 0.05)., Conclusion: The human CB CD34+ cells are able to produce functional hFIX after transduction by rAAV-2/hFIX. The cell proliferation and differentiation capacities of the host CD34+ cells were not affected by rAAV-2.

Objective: To explore whether normal platelet contains tissue factor (TF), and the significance of platelet-associated TF (PATF)., Methods: Platelets were isolated by Sepharose 2B gel column. ELISA was used to detect the TF content in the lysates of washed platelets. Procoagulant activity of PATF was measured by one stage clotting time assay. The mRNA of TF was detected by reverse transcription polymerase chain reaction (RT-PCR)., Results: A certain amount of TF antigen (16.37 +/- 6.39) ng/L was detected in the washed-platelet lysates. Upon activation by collagen, platelets released TF and caused a marked increase in TF level in plasma (P <0.05). Resting platelets had no TF procoagulant activity, while procoagulant activity of platelets activated by collagen increased significantly, which could be blocked by TF McAb and poor VII plasma. TF mRNA could not be detected in washed platelets. TF content in platelets from patients with coronary heart disease was significantly higher than that from normal controls (P < 0.05). Resting platelets from the patients showed a higher procoagulant activity, which could be inhibited by TF McAb., Conclusion: Platelets contain TF and the latter released by activated platelet was functionally active. Platelet itself might not synthesize TF. Protein content and procoagulant activity of PATF in patients with coronary heart disease were higher than that in controls. All these indicate that platelet may be involved in coagulation and thrombosis by releasing TF.

Objective: To explore the change in the protein content and procoagulant activity of platelet-associated tissue factor (TF) in thrombotic diseases., Methods: Platelets were isolated with Sepharose 2B gel column. ELISA was employed to detect the TF protein content in the lysates of washed platelets, procoagulant activity of platelet-associated TF was measured with one stage clotting time assay., Results: A certain amount of TF antigen (16.37 +/- 6.39) ng/L was detected in resting platelets in normal controls. The procoagulant activity of resting platelets and platelets activated by collagen were (0.89 +/- 0.26), (2.27 +/- 0.24) U/ml respectively. While specific TF McAb was added, the procoagulant activity of activated platelets can be markedly inhibited (1.39 +/- 0.28) U/ml (P < 0.001). It is thus proved that the procoagulant activity of activated platelets mainly comes from TF. TF protein content of the lysates of washed platelets in coronary heart disease group, brain infarction group and diabetes mellitus group was (20.71 +/- 8.78), (23.83 +/- 8.03), (22.12 +/- 8.24) ng/L respectively. The procoagulant activity of platelets without collagen treatment in these thrombotic diseases was (1.71 +/- 0.24), (1.69 +/- 0.25), (1.75 +/- 0.31) U/ml respectively. When specific TF McAb was added specific TF McAb, the procoagulant activity was (1.43 +/- 0.25), (1.39 +/- 0.25), (1.40 +/- 0.29) U/ml respectively. The TF protein content of lysates of washed platelets and the procoagulant activity of platelets without collagen treatment in patients with thrombotic diseases were significantly higher than those in normal controls. This procoagulant activity can be inhibited by TF McAb (P < 0.01)., Conclusion: The change in platelet-associated TF in thrombotic diseases may contribute to the maintenance of hypercoagulability.

Vasovagal syncope is an uncommon complication during acupuncture. However, convulsive syncope during acupuncture treatment in older individuals is seldom reported in the literature. Two older patients who experienced convulsive syncope during acupuncture treatment at Taipei Veterans Hospital, Taipei, Taiwan, from January 2000 to December 2002 are reported. These cases are instructive to acupuncturists. Although acupuncture treatment is generally safe in most situations, one needs to be cautious in delivering acupuncture to older and debilitated individuals.

The nuclear factor kappa B(NF-kappaB) plays a crucial role in inflammatory, immune response, embryo development, cell proliferation and apoptosis, cell cycle control as well as tumorgenesis. In recent years, a variety of investigations have demonstrated that NF-kappaB was closely associated with the pathogenesis of hematological malignancies such as leukemia, lymphoma and multiple myeloma. Nowadays, increasingly attention has been paid to the studies on the activation and its mechanism of NF-kappaB in the hematogenic malignancies. So that, in this article, progress on these aspects is reviewed.

Background: The purpose of this study was to examine the effects of duration of hormone therapy (HT) and treatment regimens on mammographic density., Methods: A retrospective study was carried out of of 467 post-menopausal women who received estrogen or estrogen-progestin and had regular mammographic density determination by the Breast Imaging Reporting and Data System between 1994 and 2001., Results: The fraction of women using HT who had an increase in mammographic density became more important over time. Further analysis of the effects of regimens after 4 years of HT shows that the increase in mean density was much greater in women receiving combined HT than in those receiving estrogen alone. The incidence of increased mammographic density showed significantly progressive increases over the duration of combined HT from 7.5 to 22.4%., Conclusions: Although most women using HT maintained breast density at pre-treatment levels, there is a note of caution for women using long-term HT, especially those using combined estrogen-progestin.

To explore the effect of glycosyl-phosphatidyl inositol-specific phospholipase D (GPI-PLD) on the adhesion function of bone marrow mononuclear cell from patients with myeloid leukemia and analyze its mechanism, the activity of GPI-PLD in bone marrow mononuclear cell from the patients were measured by using GPI-anchored placental alkaline phosphatase (PLAP) as substrate and Triton-X114 partitioning; the adhesion rate and CD24 expression of these cells were measured by MTT and immunohistochemical method respectively, when these cells were or were not treated by 1 mmol/L 1,10-phenanthroline for 5 hours. The results showed that the GPI-PLD activity of bone marrow mononuclear cells from the patients was significantly inhibited after being treated by 1 mmol/L 1, 10-phenanthroline for 5 hours [(42.08 +/- 7.21)% vs (5.4 +/- 2.96)%], while the adhesion rate and the expression of CD24 of these cells were increased [(49.78 +/- 26.73)% vs (61.19 +/- 29.14)%, (16.02 +/- 9.68)% vs (18.5 +/- 11.14)%, respectively)]. It is concluded that depression of GPI-PLD activity can increase the adhesion rate of bone marrow mononuclear cells from the patients while the CD24 expression is enhanced.

To investigate the effect of GFP fused to C terminal of integrin alpha(IIb) on the biosynthesis and expression of alpha(IIb) beta(3) compound, the alpha(IIb) GFP expression plamid, named palpha(IIb) GFP, the cDNA of alpha(IIb) was constructed from p3.1-2b and fused to pEGFP-N1 in frame. When the sequence of palpha(IIb) GFP was confirmed by sequencing it was transferred to Chinese Hamster Ovary (CHO) cells with or without p3.1-3a expressing integrin beta(3). Then the expression of alpha(IIb) GFP fusion protein was confirmed by Western blot and then its subcellular localization was determined with laser confocal scanning microscopy. The results showed that the target gene was cloned into recombinant vector by restriction analysis and sequencing. Overexpression of the fusion protein in the transfected CHO cells was identified with Western blot. Subcellular localization analysis confirmed that alpha(IIb) GFP was expressed in CHO cells and could be transferred from endoplasmic reticulum to Golgi apparatus. It is concluded that the eukaryotic expression plasmid containing alpha(IIb) GFP fusion gene is successfully constructed. GFP fused to the cytoplasmic tail of integrin alpha(IIb) allows the normal expression of alpha(IIb) beta(3) in CHO cells.

Objective: To study the effect of glycoprotein (GP) alpha II bA2334C mutation on the biosynthesis and expression of alpha II bbeta3 complex., Methods: The GP alpha II bA2334C eukaryotic expression plasmid pc3.1-2334M2b was constructed. Chinese hamster ovary (CHO) cells were transfected with the plasmid with or without integrin beta3 expression plasmid pc3.1-3a. The whole expression of alpha II bA2334C was confirmed by Western blot and the membrane expression was analyzed by flow cytometry. A newly constructed alpha II bA2334C GFP fusion protein expressing plasmid was used to determine its subcellular localization by laser confocal scanning microscopy., Results: Expression of the mutant protein, alpha II bA2334C, in the transfected CHO cells was confirmed by Western blot with a lower rate of the mature type than the wild type control. The expression on membrane was only 25% of the normal. Subcellular localization analysis showed that alpha II bA2334C GFP was able to be expressed in CHO cells and could be transported from endoplasmic reticulum to Golgi apparatus., Conclusions: The mutant alpha II bA2334C can be synthesized in CHO cells and form alpha II bbeta3 complex. However, only a small fraction of the premature alpha II bA2334C can be transported to Golgi apparatus and transformed to mature alpha II b. The possible pathogenesis of this type II thrombasthenia may be that the misfolded alpha II bA2334C is partially degraded in the endoplasmic reticulum causing lower expression of alpha II bbeta3 complex on the membrane and resulting in impared function of platelets than normal alpha II b.

The current therapeutic approaches for pulmonary fibrosis, which is characterized by fibroblast proliferation and extracellular matrix remodeling, are unsatisfactory. Feitai, consisting of several herbs, is a folk formula for pulmonary tuberculosis therapy in China. To investigate the effects of Feitai on pulmonary fibrosis, Feitai was administered orally to bleomycin (BLM)-treated rats, and the lung toxicity effects were evaluated according to inflammatory cell count, protein concentration, and lactate dehydrogenase (LDH) activity in the bronchoalveolar lavage fluid (BALF), malondialdehyde level and hydroxyproline content in lung tissue 28 days post-BLM. Serial sections of the lung were stained with hematoxylin and eosin (HE) and Masson trichrome, respectively. The degree of fibrosis was assessed quantitatively using LEICA QWin image analyzer. Results showed that Feitai inhibited BLM-induced lung fibrotic lesions in a dose-dependent manner as reflected by decreased the lung hydroxyproline content and lung fibrosis fraction 28 days after BLM instillation. Treatment with Feitai also significantly ameliorated the BLM-induced lung toxicity effects detected in BALF and lung tissue. The effects in vitro on WI-38 human lung fibroblast cell line showed that Feitai significantly reduced the cell proliferation and transforming growth factor (TGF)-beta stimulated type I collagen synthesis. These results strongly demonstrate that Feitai may be useful in the treatment of pulmonary fibrosis.

Background: Palivizumab (PZ) is the only monoclonal antibody in use against a human infectious disease. PZ is given as prophylaxis against infection with respiratory syncytial virus (RSV). An RSV escape mutant, MP4, has been shown to resist PZ prophylaxis in cotton rats., Methods: To further define the potential of RSV to resist prophylaxis, additional PZ-resistant viruses were selected in cell culture and were tested for susceptibility to PZ in cotton rats., Results: Mutant MS412 had an A-->C mutation at nucleotide position 827 in the F gene, resulting in an amino acid change from Lys to Gln at position 272. Mutant F212 had an A-->T mutation at position 816, leading to an amino acid change from Asn to Ile at position 268. In vitro, F212 had impaired growth kinetics. In cell culture, F212 was partially and MS412 was completely resistant to PZ neutralization. A single prophylactic dose of 15 mg/kg PZ protected cotton rats from infection with F212 but not with MS412., Conclusion: Both in vitro and in vivo, individual RSV PZ escape mutants varied in their susceptibility to PZ. Mutations associated with resistance to PZ did not always result in failure of PZ prophylaxis.

This study examines family members' caregiving gains as a result of caring for their relatives with schizophrenic spectrum disorders, and the influence of formal and informal social support on these positive experiences. The results from interviews with 560 family members support that the experience of caregiving gains is prevalent. Moreover, formal support from mental health professionals through information sharing and collaborative interactions with family members, and informal support from other family members, support group participation, and contributions from the relative with mental illness all have significant, positive associations with family members' experiences of caregiving gains.

Objective: To prepare the rAAV2/hFIX and evaluate the efficiency of the preparation on gene therapy of hemophilia B model mice., Methods: The rAAV-2/hFIX was prepared by "one helper virus-one vector cell line" strategy and transfected both BHK-21 and C2C12 cells in vitro. The hFIX antigen level in cell culture supernatant was assayed. The rAAV-2/hFIX was injected into muscles of hemophilia B model mice and assayed the serum hFIX levels, hFIX clotting activity, bleeding time, 5 min bleeding volume., Results: The hFIX antigen could be detected from 24 h till 120 h after BHK-21 and C2C12 cells were transfected with highest levels at 24 h reaching (51.0 +/- 6.5) ng/10(5) cells and (68.0 +/- 7.2) ng/10(5) cells, respectively. The rAAV2/hFIX injected mice could efficiently express hFIX and peaked at three weeks after injection, then slowly decreased but low level hFIX antigen was still detectable till 10 weeks after injection. There were significant differences between the high, middle and low dose groups of rAAV2/hFIX and the control group (P < 0.01), the plasma FIX clotting activities in the model mice were improved remarkably, bleeding time was greatly shortened and bleeding in 5 min was decreased. The hFIX expression level and FIX clotting activity of the high dose of rAAV2/hFIX group (1.6 x 10(13) v.g./kg) reached about (387.0 +/- 12.5) ng/ml plasma in contrast with the normal levels of (30.0 +/- 5.5)% at the third week after injection. No rAAV2 vector DNA was detected in the organs except for injected muscle tissue., Conclusion: The rAAV2/hFIX transfected BHK-21 and C2C12 cells could efficiently express hFIX antigen and was of therapeutic effects for the hemophilia B model mice by intramuscularly injection.The results provide the basis for clinical trial of rAAV2 gene therapy for hemophilia B.

Objective: A ruptured cornual pregnancy is a rare and challenging problem. We present two cases of cornual pregnancies after in vitro fertilization and embryo transfer (IVF-ET) treated by cornual resection, with an excellent perinatal outcome for the intrauterine pregnancy in both cases. A literature review of cornual pregnancy after IVF-ET is also included., Case Reports: Two women had undergone IVF-ET because of tubal problems. Emergent laparotomy was performed because of internal bleeding at 12 weeks of gestation in one case and 17 weeks in the other, and in both cases, ruptured cornual pregnancies were found. Cornual resection and primary repair were performed. The women were discharged on the 6th and 7th postoperative day, respectively, and they underwent an elective cesarean delivery at 37 weeks of pregnancy. They were delivered of healthy babies, one weighing 2700 g and the other 2310 g., Conclusion: These cases illustrate that good perinatal outcomes can be achieved by surgical intervention in heterotopic pregnancies, even in the event of a ruptured cornu., (Copyright 2004 International Federation of Gynecology and Obstetrics)

The aim of this study was to apply a new type piezo-electric immunosensor in immunology classification for acute leukemia. Based on the plasma-polymerized film and the nanogold particles self-assembly technology, a new type piezo-electric immunosensor was firstly developed to absorb and fix the monoclonal antibodies for detecting leukemia cell antigens in patient blood samples. The results showed that the positive rate of detection with this method was quite close to that with the fluoroimmunoassay which are commonly used in clinic work. There was no significant difference between the positive rates (chi(2) = 3.4, P > 0.05). It is concluded that this piezo-electric immunosensor detection has many advantages, such as convenient operation, real time operation and control, qualitative detection, and less cost. This method can be used for acute leukemia immunology classification.

Pulmonary fibrosis is a common consequence of numerous pulmonary diseases. The current therapeutic approaches for this condition are unsatisfactory. Feitai, a composite formula consisting of several herbs, is used in China as a folk remedy for treating patients with pulmonary tuberculosis. In this study, we extensively investigate the effects and mechanisms of Feitai on bleomycin (BLM)-induced pulmonary fibrosis in rats. One hundred and twenty male Sprague-Dawley rats were randomly divided into four groups, referred to as the saline-water, saline-Feitai, BLM-water, and BLM-Feitai groups. Following a single instillation of BLM (5 mg/kg) or saline, rats were orally administered Feitai at a dose of 3 g/kg body weight or sterilized distilled water once daily. Rats were killed at 7, 14, or 28 d post-BLM. Inflammatory cell count, protein concentration, and lactate dehydrogenase activity in bronchoalveolar lavage fluid were measured, and myeloperoxidase activity and lipid peroxide content in lung homogenates were analyzed. Treatment with Feitai inhibited lung fibrotic progression induced by BLM, as indicated by the decrease in lung hydroproline content and lung fibrosis score at 28 d post-BLM. This was accompanied by significant amelioration of BLM-induced body weight loss, lung edema, and inflammatory response during the development of lung injury in the acute phase. The results strongly indicate the beneficial effects of Feitai in protecting against BLM-induced pulmonary fibrosis. Furthermore, the inflammatory response and lipid peroxidation were inhibited by Feitai, suggesting that the effect of this formula on BLM-induced lung injury and fibrosis is associated with antiinflammatory and antioxidant properties.

Objective: To explore the dynamic change of Phlegm-stasis in the rat atherosclerotic model as the time goes on., Methods: Adopting high fat forage fed to develop the atherosclerotic model in rats, and the changes of blood lipid, hemorrheology, blood glucose, insulin and vascular smooth muscle cell (VSMC) actin expression were detected by biochemical and immunohistochemical assay at various time points after modeling., Results: The expression of VSMC actin gradually increased along with the change of model rats' Syndrome from Phlegm to stasis, i.e., the change of parameters, including blood lipid, hemorrheologic parameters, blood glucose, insulin and insulin sensitive indexes along with the aggravation of disease., Conclusion: The expression of VSMC actin could be the molecular mechanism for the Syndrome developing from Phlegm to stasis in atherosclerotic rats.

Background: Postmenopausal osteoporosis is associated with estrogen deficiency. Estrogens have effects on bone metabolism, which are mediated by estrogen receptors (ERs). If estrogen responsiveness is related to the ER expression level, ER expression in postmenopausal women should be different from previous studies using osteoblast lineage. We investigated the expression of variant isoforms of ER messenger ribonucleic acid (mRNA) in osteoblasts (OB) from postmenopausal women and a human osteosarcoma cell line, MG 63., Methods: Osteoblast cultures were prepared from the upper femur of postmenopausal patients or MG 63. For OB cultures at 5, 10, 15, 20, and 25 days, the expressions of ERalpha and beta mRNA were examined using reverse transcriptase-polymerase chain reaction., Results: In MG 63, ERbeta mRNA was constantly and highly expressed during the 25-day culture, whereas ERalpha mRNA was barely detected. In the primary OB cells, both ERalpha and beta mRNA were transcribed during the 25-day culture, but expression of ERalpha mRNA was much stronger than that of ERbeta mRNA. A splice variant form of ERbeta mRNA that was missing the entire exon 2 (ERbeta delta 2) was detected and heterogeneously expressed in OB cultures from 16 postmenopausal women., Conclusion: Differential expressions of these ER isoforms suggest that they may have different functions or that they interact with each other during bone metabolism. The different ratio of ERbeta to ERbeta delta 2 mRNA or ERalpha to ERbeta mRNA expressions in osteoblast cultures may be related to different bone conditions. Whether the presence of ERbeta delta 2 in postmenopausal women influences the biological properties of bone needs to be determined.

Palivizumab (PZ) is the only monoclonal antibody in human use against an infectious disease. PZ is a humanized monoclonal antibody that recognizes the fusion protein of respiratory syncytial virus (RSV). PZ prophylaxis reduces the likelihood of hospitalization for young children at risk for severe RSV infections. The quasispecies nature of RNA viruses allows rapid emergence of viruses with a selective advantage. A PZ resistant virus was selected by passage of RSV in the presence of PZ in cell culture. The cell culture-derived virus was completely resistant to PZ prophylaxis in cotton rats. The increasing use of PZ, and in particular, the use of PZ in immunosuppressed patients, provide opportunities for resistant viruses to emerge. Whether such viruses will appear and be of clinical significance for humans is unknown. Preclinical studies in cotton rats predicted the efficacy of PZ in humans; these results suggest that if PZ resistant viruses arise in humans, PZ prophylaxis may be ineffective.

To investigate the transduction efficiency of recombinant adeno-associated virus 2 (rAAV-2) in human umbilical cord blood CD34(+) hematopoietic stem/progenitor cells, the CD34(+) cells sorted by the method of magnetic cell sorting from human cord blood were infected with the rAAV-2 expressing the green fluorescent protein (GFP) gene. After transduction for 19 hours, the expression of GFP was detected under fluorescence microscope. The results showed that 43% CD34(+) cells expressed the GFP gene at a multiplicity of infection of 2 x 10(5). It is concluded that the rAAV-2 can transduce human cord blood CD34(+) hematopoietic stem/progenitor cells efficiently.

Objective: To establish the possible relationship between some coagulation factors and the onset of acute cerebral infarction (ACI)., Methods: The study population consisted of 71 patients with ACI confirmed by CT and 50 age-matched healthy volunteers. Blood samples were obtained during the onset period of ACI. Tissue factor (TF) and tissue factor pathway inhibitor (TFPI) activity in plasma were assayed with the chromogenic assay. Plasma TF and TFPI antigen were measured with enzyme linked immunoadsorbent assay (ELISA). Plasma F VII coagulation activity (F VII: C) and F VIII coagulation activity (F VIII: C) were developed in the one-stage system. Plasma prothrombin (FII) was determined with Ecarin assay. Plasma fibrinogen (Fbg) was measured with thrombin assay. Plasma antithrombin III activity (ATIII) was determined using heparin cofactor activity assay., Results: Compared with the control, plasma TF activity and antigen in patients with ACI were significantly higher (both P<0.05). But plasma TFPI activity and antigen were remarkably lower in the ACI group (both P<0.05). Plasma F VII: C was significantly higher (P<0.01), and F VIII: C was markedly lower (P<0.05). Plasma FII was remarkably higher (P<0.01). Similarly the Fbg was significantly higher in the ACI than that in the control group (P<0.01), whereas ATIII was significantly lower (P<0.01)., Conclusion: The initiation of TF pathway is contributed to the onset of ACI and the blood is in hypercoagulable state during the early period of ACI.

Objective: To elucidate the effect of angiotensin II (AngII) on the expression of tissue factor (TF) by monocytes and its mechanisms., Methods: Monocytes were isolated from healthy volunteers by Ficoll-Hypaque gradient and Percoll, and cultured in RPMI-1640. Procoagulant activity (PCA) was determined by one-stage clotting method, TF antigen by ELISA. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the TF gene mRNA. The levels of IkappaBalpha was detected by Western blot. Electrophoretic mobility shift assays (EMSA) were performed to evaluate the activity of NF-kappaB., Results: AngII (10(-9) - 10(-7) mol/L) significantly increased monocyte PCA, TF antigen and TF mRNA expression in a dose and time dependent manner. Losartan (10(-6) - 10(-5) mol/L) significantly inhibited the effects of AngII on TF activity, antigen and mRNA expression in a dose-dependent effects. Staurosporine (2.5 x 10(-7) mol/L) and genistein (4 x 10(-5) mol/L) lowered TF level of monocytes (P < 0.05). Western blot analysis revealed that after exposure to AngII (10(-7) mol/L), IkappaBalpha level decreased at 15 min, reached nadir at 60 min, and recovered at 180 min. EMSA showed NF-kappaB binding activity increased at 15 min, reached peak at 60 min, and recovered at 180 min. Pyrrolidine dithiocarbamate (PDTC, 10(-4) mol/L), an inhibitor of NF-kappaB, or AT1R antagonist losartan (10(-5)mol/L) inhibited AngII-induced NF-kappaB translocation., Conclusions: AngII could induce the expression of TF in human monocytes, and this effect was mediated by AT1R. The PKC pathway played the most important role in AngII-induced TF expression. The activation of NF-kappaB was involved in TF expression in monocytes.

Erythropoietic protoporphyria (EPP) is characterized by excess accumulation of protoporphyrin, which is due to deficient activity of the enzyme ferrochelatase (FECH). This results in photosensitivity and in some patients liver disease which may necessitate liver transplantation. The aim of this study was to delineate the abnormalities in the FECH gene which cause phenotypic expression in EPP. We identified 43 individuals from 25 North American families with EPP who were heterozygous for various FECH mutations, but the mutations did not adequately explain the variable phenotype. We also examined the presence of an intron polymorphism (IVS3-48c) in the FECH gene which was shown to cause the formation of aberrantly spliced FECH mRNA. FECH DNA analysis demonstrated that 94% of 31 symptomatic individuals with FECH mutations were heterozygous for IVS3-48c, whereas 12 asymptomatic individuals with FECH mutations were homozygous for IVS3-48t. Haplotype analysis in four families showed that symptomatic members had the IVS3-48c polymorphism in the non-mutant FECH allele. Sequencing of the proximal FECH gene promoter showed no additional changes which might affect gene expression. The levels of normal FECH mRNA, measured by relative quantitative RT-PCR, and FECH enzyme activity were correspondingly lower in the cultured lymphoblasts of family members with the IVS3-48c polymorphism. These results indicate that symptomatic disease in most North American patients with EPP is explained by the inheritance of a mutation in one FECH allele which causes a structural alteration in the protein, together with a low expressing non-mutant FECH allele which is caused by the IVS3-48c polymorphism.

To study the relationship between the level of the soluble L-selectin (sL-selectin) in plasma and surface L-selectin expression on leukemic cells and episode and state of illness in acute leukemia patients, the plasma level of sL-selectin was measured by a sandwich enzyme-linked immunosorbent assay, and the expressions of surface L-selectin and its gene (lyam-1) were detected by immunohistochemistry and RT-PCR. The results showed that the levels of sL-selectin were significantly higher in untreated and therapy-resistant acute leukemia patients, and expression of L-selectin mRNA and cell surface L-selectinin in untreated and NR patients were significantly lower than that in CR patients and control group (P < 0.05). The plasma levels of sL-selectin were significantly increased in patients with splenomegaly and hepatomegaly (extramedullary infiltration). The levels of sL-selectin were related to the clinical course of the acute leukemia patients. A significant correalation existed between expressions of L-selectin mRNA and surface L-selectin in acute leukemia patients (gamma = 0.782, P < 0.05). It is concluded that expression of L-selectin gene was down-regulated in level of mRNA and protein in acute leukemia patients and both changes were highly correlated. Monitoring of the plasma level of sL-selectin is possibly useful for early diagnosis of relapse and extramedullary infiltration in acute leukemia.

Objective: To study the signal transduction in the course of energy metabolism of platelets, and to provide a new theoretical basis for clinical application of the anti-activation of platelets by the monoantibody of integrin GPIIb/IIIa., Methods: With or without the block of integrin GPIIb/IIIa monoantibody 10E5 and/or integrin alpha v beta 3 monoantibody 609, the uptake rate of tritium-labeled 2-deoxyglucose([3H]2-DG) by platelets activated by thrombin was counted in a liquid scintillation counter. The relationship between the energy metabolism of platelets and platelet integrin was investigated by the course of glucose uptake of individual vivo platelets., Results: Compared with unactivated platelets, the glucose uptake of activated platelets increased, and it could be blocked by monoantibody 10E5 and partly blocked by 609., Conclusion: The glucose uptake of activated platelets can be suppressed by monoantibody 10E5, and the intergrin family is associated with the signal transduction of platelet energy metabolism. Integrin GPIIb/IIIa may play an important role in the procedure of platelet energy metabolism.

Objective: To compare the different effects of the programmable cell freezer and the -70 degrees C mechanical freezer in the preservation of the combilical cord blood and to study the effects of magnetic activated cell sorting on cell biologic characters., Methods: After separating the nucleated cells from the umbilical blood by the density gradient separation method, we used cryoprotectant to protect the nucleated cells and preserve them by two methods; one was using the programmable cell freezer to cool the cells and finally putting them to the liquid nitrogen, and the other one was directly preserving the cells in a -70 degrees C mechanical freezer. The cells were thawed after a month and tested for the viability and clonogenic ability. Cell recovery, clonogenic capacity and clonogenic recovery by the two cryopreservation methods were compared. Meanwhile, the CD34+ cells were sorted by the method of magnetic cell sorting from the umbilical cord blood, and the cell purity and recovery after sorting were analyzed by the method of fluorescence-activated cell sorting., Results: There were no significant differences between the two preserving methods in cell viability, cell recovery, clonogenic capacity, or colony recovery. The purity of the CD34+ cells was 88.3% and the recovery rate was 46.2% after the cell sorting. The cell viability and clonogenic capacity of the CD34+ cells were much higher than those of the CD34- cells., Conclusion: The umbilical cord blood can be efficiently preserved by the two methods and highly purified CD34+ cells could be obtained by using the method of magnetic cell sorting. The procedure of cell sorting doesn't affect the viability and clonogenic ability. The study provides some evidence for clinical application of preserving the umbilical cord blood and transplanting CD34+ cells.

Objective: To evaluate the efficacy of hematopoietic stem cell transplantation (HSCT) in hematological malignancies., Methods: Twenty-five patients with hematological malignancies were treated by autologous bone marrow transplantation (ABMT), autologous peripheral blood stem cell transplantation (APB-SCT) or Allo-PBSCT. Eight acute myeloid leuekmia (AML) patients who received autologous hematopoietic stem cell transplantation (AHSCT) were treated by chemotherapy again., Results: All the patients engrafted gained hematopoietic reconstitution; they were followed up for 6 to 91 months; five patients died; twenty patients were alive in a disease-free situation., Conclusion: After the patients with acute leukemia receive AHSCT, the relapse incidences can be reduced obviously by regular chemotherapy. HSCT is one of the best methods for hematological malignancies.

Objective: To investigate the effect of the defect of integrin alpha II b beta 3 on the inside-out signal transduction in platelets., Methods: The transfected cDNA, its expression and the ability of cells binding to PAC-1 and fibrinogen were investigated by RT-PCR, DNA sequence analysis, flow cytometry and Western blotting., Results: The integrin alpha II b beta 3 level in the patients with Glanzmann's thrombasthenia was significantly lower than that of the normal subjects and the platelets of the patients failed to bind PAC-1 activated by ADP. There were both C3077G and G3078C mutations in exon 30 of mutant alpha II bR995A beta 3 cDNA, which resulted in an amino acid substitution arginine (R) 995 to alanine (A). CHO cells transfected with wild-type alpha II b beta 3 or mutant alpha II bR995A beta 3 cDNA respectively expressed normal alpha II b beta 3 and mutant alpha II bR995A beta 3. When cells were not activated, wild-type alpha II b beta 3 CHO cells failed to bind PAC-1, but could adhere to fibrinogen, but mutant chimera alpha II bR995A beta 3 CHO cells were able to bind PAC-1 and exhibited high affinity binding fibrinogen., Conclusion: The point mutation in integrin alpha II bR995A can induce the inside-out signal transduction in platelets, and have integrin alpha II bR995A beta 3 locked into an activation state; the defect of alpha II b beta 3 in patients with GT impairs the inside-out signal transduction mediated by alpha II b beta 3 in platelets and the adhesive functions of platelets.

Objective: To evaluate effects of 17beta-E(2) and 1alpha,25(OH)(2)-vitamin D(3) on human osteoblast-like (hOB) cells., Design: Controlled, experimental study., Setting: University hospital., Patient(s): hOB cell cultures were prepared from the upper femur of postmenopausal patients undergoing bipolar endoprosthesis arthroplasty for a fractured femoral neck., Intervention(s): hOB cells were subcultured with either 17beta-E(2) or 1alpha,25(OH)(2)-vitamin D(3), or both., Main Outcome Measure(s): Cell proliferation and activity of alkaline phosphatase, osteocalcin, and interleukin-6., Result(s): 17beta-E(2) significantly reduced interleukin-6 and osteocalcin to 34% and 60% of control value but induced alkaline phosphatase and cell proliferation to 183% and 150% of control value. 1alpha,25(OH)(2)-vitamin D(3) significantly decreased cell proliferation to 88% of that of control group, but 1alpha,25(OH)(2)-vitamin D(3) plus 17beta-E(2) showed no difference from the control group. Alkaline phosphatase and osteocalcin were significantly increased by 1alpha,25(OH)(2)-vitamin D(3) alone or combined with 17beta-E(2), to 169% and 198% and to 144% and 144% of control value, respectively. 1alpha,25(OH)(2)-vitamin D(3), with or without 17beta-E(2), decreased interleukin-6 levels to 27% and 38% of control group, respectively., Conclusion(s): 17beta-E(2) and 1alpha,25(OH)(2)-vitamin D(3) have effects on osteoblasts. The prevention of osteoporosis by estrogen may be related not only to direct effects on osteoblastic activity and proliferation but also to indirect effects on osteoclasts by the decrease of interleukin-6 secretion.

A deficiency of ferrochelatase (FECH) activity underlies the excess accumulation of protoporphyrin that occurs in erythropoietic protoporphyria (EPP). In some patients, protoporphyrin accumulation causes liver damage that necessitates liver transplantation. The purpose of this study was to determine if specific mutations in the FECH gene are present in patients who develop liver disease. FECH cDNA and all 11 exons and their flanking intron regions in the FECH gene were amplified and sequenced by specific polymerase chain reactions. Gene mutations were determined in 34 individuals from 24 families: 14 had liver disease, 10 necessitating liver transplantation. All individuals were heterozygous for mutations that altered the coding region of FECH mRNA. The mutations in patients with liver disease were heterogenous, but usually caused a major structural alterations in the FECH protein, most commonly as a result of exon skipping in FECH mRNA. However, the mutations could not account for the severe phenotype by themselves, since the same mutations were found in asymptomatic family members of patients with liver disease and in patients from families in which liver disease was not present. Other genetic factors, and possibly acquired factors, also must be critical to the development of this severe phenotype in EPP.

We report on the prenatal diagnosis of ring chromosome 15 in a fetus with increased nuchal fold and intrauterine growth restriction (IUGR). A 27-year-old woman gravida 2, para 1 had normal maternal serum screen tests in the early second trimester of the index pregnancy. Fetal nuchal fold thickening up to 8 mm was incidentally found during the routine obstetric ultrasound scan at 20 weeks' gestation. Amniocentesis was undertaken and the fetal karyotype was found to be 46,XY,r(15) on cytogenetic study. Fluorescence in situ hybridization (FISH) using a telomeric probe of chromosome 15 demonstrated a terminal deletion on the q arm of the ring-shaped chromosome 15. This is the first report of a prenatally diagnosed case of ring chromosome 15. Moreover, nuchal fold thickness in the second trimester may have a role in its prenatal diagnosis., (Copyright 2001 John Wiley & Sons, Ltd.)

Objective: To determine the occurrence of activated protein C resistance (APCR), to identify APCR is associated with thrombotic events (TEs), and acquired APCR is associated with the presence of antiphospholipid antibodies (APLAs) in 30 patients with systemic lupus erythematosus (SLE)., Methods: Laboratory tests included dilute Russell's viper venom time assay for LA (dRVVT-LA), ELISA assay for ACL, APC sensitivity ratio, and factor V Leiden were detected by PCR-Mnl/I digestion., Results: Acquired APCR was presented in 14(46.67%) of 30 patients. Factor V Leiden was not found in any patients. The incidence of TEs in the APCR-positive patients was significantly higher than that in the APCR-negative patients (42.85% vs 6.25%, P < 0.05). The incidence of TEs in the LA-positive patients was also significantly higher than that in the LA-negative patients (50% vs 11.1%, P < 0.05)., Conclusions: The presence of either APCR or LAs is associated with one of the risk factors of TEs (P < 0.05). There is not a significant interaction between APCR and LAs in the association with TEs. Acquired APCR may not reflect the interference of LAs with the protein C pathway which may represent a mechanism of LA-associated TEs.

Objective: To determine the effects of oral and transdermal hormone replacement therapy on lipid profile and hemostatic factors in postmenopausal women., Design: Twenty subjects were treated with oral E2 valerate (2 mg) combined with cyproterone acetate (1 mg) (group I) and 21 with transdermal E2 (1.5 mg) plus oral medroxyprogesterone acetate (5 mg) (group II). The effects on lipid profile and hemostatic parameters were evaluated at baseline and after 3, 6, and 12 months of treatment., Results: Group I showed a stronger increase of high-density lipoprotein (HDL) cholesterol levels (2-8%) and stronger reduction of atherogenic indices (total cholesterol/HDL cholesterol and low-density lipoprotein/HDL cholesterol) than group II. Group II showed a more pronounced reduction of triglyceride (21-31%) and factor VII (6-10%) levels than group I. Both groups showed reduced concentrations of total cholesterol, low-density lipoprotein cholesterol, tissue plasminogen activator, plasminogen activator inhibitor-1, antithrombin III, and protein S, whereas protein C was increased after 12 months of treatment., Conclusions: The cardioprotective effects of hormone replacement therapy are demonstrated by favorable effects on lipid profile and fibrinolytic activity. Oral hormone replacement therapy showed a more prominent effect on lipoprotein metabolism than did transdermal administration, but transdermal medication had a stronger effect on triglyceride and coagulation factors. However, it needs to be considered that there is an increased risk of venous thrombotic events in the first year of treatment.

We performed a positron emission tomography study, using regional cerebral blood flow as the index of brain activity, to address the specificity of brain activation pattern by acupuncture stimulation of short duration at the classical analgesic point. Needling manipulation at 2 Hz was performed at a classical point of prominent analgesic efficacy (Li 4, Heku) and a near-by non-classical/non-analgesic point, respectively, in normal subjects. Regions activated by acupuncture stimulation at Li 4 included the hypothalamus with an extension to midbrain, the insula, the anterior cingulate cortex, and the cerebellum. Of note, it was only the stimulation at Li 4 that activated the hypothalamus under the similar psychophysical ratings of acupuncture sensation (deqi) as elicited by the stimulation at the two points, respectively. The data suggested that the hypothalamus might characterize the central expression of acupuncture stimulation at the classical analgesic point and serve as one key element in mediating analgesic efficacy of acupuncture stimulation.

Background: The beneficial effect of hormone replacement therapy (HRT) on cardiovascular disease has been documented in postmenopausal women, but has a significant time trend. Thus, it is worthwhile to further study whether there are different effects on cardiovascular factors between short- and long-term use of HRT., Method: Prospective study of the changes on lipoprotein profile, hemostatic factors, and platelet aggregation was evaluated in 21 postmenopausal women receiving oral E2 valerate (2 mg/d) combined with medroxyprogesterone acetate (10 mg/d) during the last 10 days of each 21-day cycle. The treatment period was 24 months., Results: During the 24 months of treatment, total cholesterol, low-density lipoprotein cholesterol (LDL-C), and atherogenic indices- total cholesterol-to-high-density lipoprotein cholesterol (HDL-C) and LDL-C-to-HDL-C, were significantly reduced. The concentrations of tissue plasminogen activator and plasminogen activator inhibitor-1 were significantly reduced after 12 months of HRT. In addition, the concentrations of antithrombin III were significantly increased, but protein S was statistically decreased during the 18 months of HRT. The maximum aggregation and slope of platelet aggregation were significantly reduced only during the first 12 months of HRT., Conclusion: This study demonstrates that there were some differences in cardiovascular risk factors between short- and long-term HRT, especially in changes in platelet aggregation and hemostatic factors. However, the long-term favorable effect on lipoprotein metabolism and fibrinolytic activity among hormone users may explain, in part, the inverse association between HRT and cardiovascular disease.

Myofascial pain syndrome (MPS) in the cervical and upper back regions is a common medical problem. The involved muscles include trapezius, multifidi, splenius cervicis, levator scapulae, supraspinatus or infraspinatus. Acupuncture is a well-known method for relief of chronic pain. In this study, we evaluated the effect of acupuncture in MPS and its durability by using the concept of meridians. Twenty-nine patients with chronic MPS in the upper back and cervical regions received acupuncture 2 times per week for 3 weeks. According to the pathways of the meridians, we chose acupuncture points by the pain regions located in which parts of meridian passing (i.e. "Where the meridian passes, where to treat the disease."). We evaluated the intensity of pain by visual analog scales (VAS) and active range of motion (ROM) of neck before and after therapy. After receiving acupuncture for 3 weeks, VAS of pain fell significantly from 9.0+/-1.0 to 4.9+/-2.5 and active ROM of neck significantly increased from 35.8+/-10.2 degrees to 61.3+/-9.4 degrees (p<0.05). The onset of symptom relief was gradual and duration of symptom relief after a course of acupuncture was average 5.3+/-1.2 days. In conclusion, acupuncture is a somewhat effective method for pain relief of patients with chronic MPS in the cervical and upper back regions. However, the effect of acupuncture with the concept of meridians on MPS is insidious and the duration of the relief is not long enough.

Unlabelled: RT-PCR method was used to detect the expression level of bcl-2 gene in 38 patients of acute leukemia., Results: Expression level of bcl-2 in the patients with clinical refractoriness increased more obviously than previously untreated and CR patients, and the expression level of bcl-2 was obviously correlated with clinical refractoriness(r = 0.83, P < 0.01), which indicated that high expression of bcl-2 was one of the factors in clinical refractoriness. It was also a sensitive method for predicting the effect of anti-tumor agents and prognosis in clinic work.

A 77-year-old woman was referred for Ga-67 scan to evaluate intermittent fever and chills that had lasted more than 20 days. The Ga-67 whole-body scan revealed a doughnut-shaped Ga-67 accumulation in the lower abdominal region. Combined Ga-67 and Tc-99m MDP bone scan confirmed that this activity was in the uterus, because the shape of the urinary bladder on bone scan was different from that of the Ga-67-avid lesion. Pyometra was proved during operation, and pus culture was performed.

A series of 2,3-disubstituted-4-phenylquinolines were prepared and were found to inhibit the apical sodium co-dependent bile acid transporter (ASBT). Alkyl and ester substitution at the 3-position showed comparable activities while substitution at the 2-position was much more sensitive to the nature of the substituent. The synthesis and in vitro potency data are presented for this novel class of compounds.

Presacral neurectomy is an effective treatment for chronic pelvic pain and dysmenorrhea. The theory behind presacral neurectomy for pain relief is based upon the anatomy of the sensory pathways from the pelvic viscera through the inferior and superior hypogastric plexus, located in the presacral area, to the spinal columns. The excision of the presacral nerve trunk can obstruct pain sensory pathways. Unfortunately 20-25% of patients treated medically for severe, disabling pelvic pain fail to show improvement. As a result, presacral neurectomy has become an alternative management for those with chronic pelvic pain and dysmenorrhea, especially for those who have failed to respond to medical treatment. Operative endoscopy further facilitates the adaptation of the laparoscopic approach to this pelvic denervation. The efficacy of pain relief by laparoscopic presacral neurectomy in chronic pelvic pain and dysmenorrhea is documented at about 75-80%. Major complications are rare with a rate of less than 1% reported. Laparoscopic presacral neurectomy therefore offers an effective alternative treatment and should also be considered as the initial surgical intervention for chronic pelvic pain.

Modulation of voltage-dependent transient K(+) currents (A type K(+) or K(A) current) by Zn(2+) was studied in rat hippocampal neurons by the whole-cell patch-clamp technique. It is found that Zn(2+) selectively binds to the resting (deactivated or closed) K(A) channels with a dissociation constant (K(d)) of approximately 3 microM, whereas the affinity between Zn(2+) and the inactivated K(A) channels is 1000-fold lower. Zn(2+) therefore produces a concentration-dependent shift of the K(A) channel inactivation curve and enhances the K(A) current elicited from relatively positive holding potentials. It is also found that the kinetics of Zn(2+) action are fast enough to compete with the transition rates between different gating states of the channel. The rapid and selective binding of Zn(2+) to the closed K(A) channels keeps the channel in the closed state and explains the ion's concentration-dependent slowing effect on the activation of K(A) current. This in turn accounts for the inhibitory effect of Zn(2+) on the K(A) current elicited from hyperpolarized holding potentials. Because the molecular mechanisms underlying these gating changes are kinetic interactions between the binding-unbinding of Zn(2+) and the intrinsic gating processes of the channel, the shift of the inactivation curve and slowing of K(A) channel activation are quantitatively correlated with ambient Zn(2+) over a wide concentration range without "saturation"; i.e., The effects are already manifest in micromolar Zn(2+), yet are not saturated even in millimolar Zn(2+). Because the physiological concentration of Zn(2+) could vary over a similarly wide range according to neural activities, Zn(2+) may be a faithful physiological "fine tuner," controlling and controlled by neural activities through its effect on the K(A) current.

Acardiac acephalus is a rare occurrence resulting from extensive anastomoses between the vessels of the monochorionic twins which invariably leads to death of the "perfused" twin and an estimated 50% perinatal mortality rate in the "pump" twin, due mainly to premature delivery or congestive heart failure. Once diagnosis has been documented, blocking the vascular flows between the twins has been the principle goal of treatment with the assumption that a more favorable chance of survival in the pump twin can be achieved. However, hasty surgical intervention without a well-designed preoperative evaluation may result in unfavorable outcomes. A 21-year-old nullipara was found to carry an acardiac parabiotic twin in a monochorionic twin pregnancy at 28 weeks of gestation and was referred to us from a local clinic. Expectant management based on the close monitoring of the well-being of the healthy co-twin is presented in this article along with review of published works on the management of this rare condition.

Laparoscopic presacral neurectomy is a safe, effective, and well-established surgical procedure to relieve intractable dysmenorrhea and chronic pelvic pain. In one woman, substantial lymphatic leakage occurred due to damaged lymphatic vessels. Adequate exposure and coagulation of the presacral lymphatic zone through the laparoscope resolved the problem, and a substantial amount of chylous ascites was removed. The patient fully recovered after the intervention.