Effects of vascular endothelial growth factor (VEGF) and chondroitin sulfate A on human monocytic THP-1 cell migration.

Angiogenesis serves as a crucial factor in disease development and progression, such as cancer metastasis, and monocyte migration is one of the key steps for angiogenesis. Therapeutic modulation of angiogenesis is a promising new therapeutic avenue under investigation. In this study, effects of vascular endothelial growth factor (VEGF) and chondroitin sulfate A on monocyte migration were investigated. Human monocytic THP-1 cells were from Riken Cell Bank (Tsukuba, Japan) and vascular endothelial cells (VECs) were obtained from swine thoracic aorta. The migration experimental system was adapted from Falcontrade mark Cell Culture Inserts with pore sizes of 3 and 8 microm cultured endothelial cells or not on the insert polyethylene terephthalate (PET) membranes. Four VEGF concentrations (0, 10, 50 and 100 ng/ml) and three concentrations of chondroitin sulfate A (0, 1.25 and 5.0 mg/ml) were used to investigate their effects on THP-1 cell migration ability through PET membranes and VECs monolayer. The THP-1 cell migration was evaluated by counting the number of migrated cells related to the total number of cells under a microscope. We counted the migration cells every 1 h on a Tatai-type hemocytometer using an inverted microscope for total 7 h. For inserts with pore sizes of 3 and 8 microm, the THP-1 cell migration increased with VEGF concentrations; however, cell migration decreased with the chondroitin sulfate A concentration. Our results demonstrated that VEGF accelerated monocyte migration through endothelial monolayer and chondroitin sulfate A is an effective inhibitor of monocyte migration for angiogenesis.

The role of STAT1 in activation of IL-3- and IL-5-induced eosinophils by interferon gamma.

Tyrosine phosphorylation of STAT1alpha in eosinophils after IFN-gamma stimulation has been shown, but the biological significance of eosinophil STAT1alpha activation in transmitting the signals through the IFN-gamma receptor remains unknown. The purpose of this study is to determine whether STAT1 is involved in the regulation of eosinophils by IFN-gamma-IFN-gamma receptor interaction. rhIL-3- and rhIL-5-induced eosinophils from CD34+ cells of cord blood on day 28 of culture were used. The cells were washed and further incubated in IL-3- and IL-5-free medium for 48 h. The induced eosinophils constitutively expressed CD69 and lost this expression after a further 48-hour incubation without the cytokines. IFN-gamma significantly upregulated CD69 expression on the 48-hour incubated cells. In inhibitory experiments on STAT1, a phosphorothioate oligo antisense DNA against STAT1alpha was added to IL-3- and IL-5-containing medium from day 15 to day 28 of culture. The oligo DNAs altered neither the expressions of myeloid cell marker CD9 and 13 nor the expression of IFN-gamma receptor on the cells. The added STAT1alpha antisense, but not sense, DNA significantly reduced STAT1alpha mRNA expression in the cells. The STAT1 antisense also significantly inhibited IFN-gamma-induced CD69 expression on the 48-hour incubated eosinophils. In conclusion, these results indicate that IFN-gamma induces CD69 expression in the induced eosinophils through STAT1alpha, suggesting that STAT1alpha may play a significant role in eosinophil regulation by IFN-gamma.

The role of the cytoskeleton in migration and proliferation of a cultured human gastric cancer cell line using a new metastasis model.

Using an in vitro model for investigating the mechanism of migration and proliferation of a cultured human gastric cancer cell line which we established recently, we studied the suppressive effect of inhibitors of the cytoskeleton proteins, actin and myosin, on the migration and proliferation of cancer cells. These inhibitors suppressed the capacity of cancer cells to migrate and proliferate dose-dependently. Thus the integrity of the cytoskeletal system may play an important role in the mechanism of metastasis of gastric cancer cells.

A new model to study repair of gastric mucosa using primary cultured rabbit gastric epithelial cells.

The process of wound repair was investigated using primary cultured rabbit gastric mucosal cells. A confluent monolayer gastric mucosal cell sheet consisting mainly of mucous cells was wounded to make a cell-free area of constant size. The changes in the cell-free area were analyzed quantitatively by image analysis. The wound recovered in 36-48 h in controls; wound repair was accelerated by the addition of fetal calf serum (FCS) and hepatocyte growth factor (HGF) to the medium and was retarded by inhibitors of cytoskeletal proteins. In the process of normal wound repair, 5-bromodeoxyuridine (BrdU)-positive cells appeared around the wound in 24-36 h but disappeared after complete repair. In the FCS- and HGF-treated group, BrdU-positive cells were mainly detected 12-24 h after wounding. In this model the wound was repaired in two steps: an initial cell migration stage and a later proliferation stage. In conclusion, FCS and some growth factors accelerate wound repair with the induction of both epithelial cell migration and proliferation. The cytoskeletal system plays an important role in normal gastric restoration.

[Anesthesia for left ventricular aneurysmectomy with concommittant coronary artery bypass grafting--a case of unstable angina due to infantile periarteritis nodosa].

Infantile periarteritis nodosa is a rare disease characterized by systemic obstructive arteritis involving coronary artery. This report is anesthetic experience of a 12-years-old boy with unstable angina secondary to infantile periarteritis nodosa, who underwent left ventricular aneurysmectomy and coronary artery bypass grafting. Anesthesia was induced with fentanyl, diazepam and enflurane, and transesophageal echocardiography (TEE) was used as a cardiac function monitor. The following two intraoperative crises occurred in succession: first, aggravation of myocardial ischemia followed by ventricular tachycardia, and second, sudden interruption of cardiopulmonary bypass due to blood coagulation in membrane oxygenator. In these situations, we could detect his critical situations promptly, because segmental wall-motion abnormality and global enlargement of both ventricles were observed with TEE before the onset of these crises. In these crises, we could treat him appropriately. Postoperative course was uneventful and he had no neurological deficit nor angina. We conclude that TEE is a powerful non-invasive monitor for early detection of cardiac dysfunction in heart surgery.

[Anesthetic management for cesarean section and clipping of aneurysm in a pregnant woman with ruptured cerebral aneurysm].

We report an anesthetic management for Cesarean section and clipping of aneurysm in a pregnant woman with a subarachnoidal hemorrhage secondary to a ruptured cerebral aneurysm. Anesthesia was induced with thiopental, vecuronium and sevoflurane, and maintained with sevoflurane (0.5-1%) before the delivery. Apgar score at 1 min was 1, but after 30 min her activity became almost normal. After the delivery, anesthesia was maintained with fentanyl for clipping surgery, because sevoflurane may inhibit uterine contraction and increase bleeding from the uterus after the delivery. We also infused prostaglandin E1 continuously at a rate of 50-70 ng.kg-1.min-1 to control arterial blood pressure and to maintain good contraction of the uterus during clipping surgery. We conclude that prostaglandin E1 is useful as a vasodilator after delivery because prostaglandin E1 contracts the uterus.

[YAG laser contact therapy of ovarian endometriosis under laparoscopy].

Thirty-six patients with ovarian endometriosis were treated with Nd:YAG laser contact irradiation under laparoscopic control. Indications for laparoscopy were infertility (n = 20) and dysmenorrhea (n = 16). The laparoscopic procedures so far undertaken in our clinic include: Aspiration of chocolate cyst, removal of ovarian endometriosis, adhesion-lysis, uterine nerve ablation, coagulation of peritoneal endometriosis and irrigation. No complications were seen. After undergoing this procedure, eight of 20 patients achieved pregnancy and 15 of 16 patients obtained pain relief. Serum CA125 levels were significantly decreased postoperatively. We confirmed that contact irradiation with a cone-shaped sapphire probe provides adequate incision and lysis at lower power levels, and that this method is an effective treatment for ovarian endometriosis.

Nd:YAG laser laparoscopy for ovarian endometriomas.

Thirty patients with ovarian endometriomas were treated with Nd:YAG laser contact irradiation via laparoscopy. The indications for laparoscopy were infertility (16) and dysmenorrhea (14). The laparoscopic procedures undertaken so far at our clinic were aspiration of chocolate cysts, removal of ovarian endometriomas, adnexal adhesiolysis, uterine nerve ablation, coagulation of peritoneal endometriosis and irrigation. No complications occurred. After undergoing this procedure, 6 of 16 patients achieved pregnancy, and 13 of 14 obtained pain relief. The serum CA-125 levels were significantly decreased postoperatively. We confirmed that contact irradiation with a cone-shaped sapphire probe provides an adequate incision and lysis at low power levels and that this method is an effective treatment for ovarian endometriomas.

Minimal and mild endometriosis. Nd:YAG laser treatment and changes in prostaglandin concentrations in peritoneal fluid.

Among many patients, infertility originates with minimal and mild endometriosis. Recently, peritoneal fluid factors have come to be associated with infertility in minimal and mild endometriosis patients, and prostaglandin (PG) is suspected of being one of those factors. We use the Nd:YAG laser for treatment of endometriosis; a total of 52 cases of minimal and mild endometriosis have been treated so far with the laser under laparoscopy in our clinic. At the same time we measured the peritoneal fluid volume and PG concentration in peritoneal fluid. Peritoneal fluid volume and PGE2 concentrations were found to be significantly higher than those in the control group (tubal obstruction patients). Seven of 52 patients received second-look laparoscopy three months after Nd:YAG laser treatment; the PG concentration showed a tendency to decrease in many patients, and the PGE2 concentration turned out to be insignificant. Within one year, 30 patients achieved pregnancy. In minimal and mild endometriosis patients Nd:YAG laser treatment may alter the PG concentration in peritoneal fluid, thereby increasing the change of pregnancy.

Ovarian wedge resection with contact Nd:YAG laser irradiation used laparoscopically.

Twelve women with polycystic ovary disease were treated with Nd:YAG laser contact irradiation using a cutting-type probe. Bilateral ovarian wedge resection was performed in every case. After undergoing the procedure, seven patients achieved pregnancy, and another three experienced the return of a normal menstrual cycle. Serum testosterone and androstenedione levels were decreased postoperatively, while luteinizing hormone (LH), follicle stimulating hormone (FSH) and estradiol were increased. It seems that the decrease in intraovarian androgens might have reduced the negative feedback system and thus might have increased the secretion of gonadotropin and estradiol, which in turn promoted ovulation induction and pregnancy.