Endothelin-1 production by human synoviocytes.

Immunoreactive (ir)-endothelin (ET)-1 concentrations in serum samples and synovial fluids from patients with rheumatoid arthritis were higher than concentrations in sera obtained from healthy volunteers. No significant difference in ir-ET-1 concentrations in synovial fluid was observed between rheumatoid arthritis patients and osteoarthritis patients. Cultured fluids of synovial cells collected from synovial tissues and leucocytes from synovial fluids of rheumatoid arthritis patients were studied to determine the origin of ir-ET-1 in synovial fluids. Ir-ET-1 was detected in the cultured fluids of synovial macrophage-like type A cells, but not in those of fibroblast-like type B cells from the synovial tissues or leucocytes from the synovial fluids. Longitudinal studies showed that the ir-ET-1 concentration in the cultured fluid reached a peak around 24 h after starting the culture. ET-1 secreted from macrophage-like synoviocytes may be involved in the pathogenesis of inflammatory arthritis.

Thrombin in the synovial fluid of patients with rheumatoid arthritis mediates proliferation of synovial fibroblast-like cells by induction of platelet derived growth factor.

OBJECTIVE: To investigate the clotting and fibrinolytic activities in synovial fluid (SF) from patients with rheumatoid arthritis (RA) and to examine the role of thrombin in synovial hyperplasia. METHODS: We measured the amounts of thrombin-antithrombin-III complex (TAT), antithrombin-III (AT-III), thrombin, plasminogen, alpha 2-plasmin inhibitor (alpha 2-PI), and plasmin-alpha 2-antiplasmin complex (PAP) in SF of 20 patients with RA and 16 patients with osteoarthritis (OA). The proliferative response of synovial fibroblast-like cells to thrombin was measured using [3H] thymidine incorporation. Expression of platelet derived growth factors (PDGF) in conditioned medium was analyzed using a Western blot method, and expression of the mRNA of PDGF and their receptors was analyzed by reverse transcription polymerase chain reaction. RESULTS: The amounts of clotting factors (TAT, AT-III) and fibrinolytic factors (plasminogen, alpha 2-PI, and PAP) were significantly higher in the patients with RA than in patients with OA (p < 0.01). Moreover, SF thrombin concentrations of patients with RA correlated significantly with erythrocyte sedimentation rates (rs = 0.751, p < 0.01) and serum C-reactive protein concentrations (rs = 0.531, p < 0.05). Thrombin exhibits mitogenic activity toward synovial fibroblast-like cells in vitro, and this mitogenic activity is associated with an increase in the expression of mRNA of both PDGF-alpha receptor and PDGF-beta-receptor. CONCLUSION: The high levels of thrombin activity in the SF of patients with RA and strong mitogenic activity of thrombin toward the synovial fibroblast-like cells suggest that thrombin plays an important role in the pathogenesis of RA.

Activation but not inactivation of the MTS1 gene is associated with primary colorectal carcinomas.

Several investigators have reported that alterations of the multiple tumor suppressor 1 (MTS1) gene frequently occurred in certain types of tumors. We examined the expression levels and somatic mutations of the MTS1 gene in primary colorectal carcinomas. The MTS1 transcript was quantitatively detected in 14 of 17 tumors, but in only one case of adjacent normal mucosa. Direct sequencing of the amplified MTS1 gene showed no somatic mutation in 17 primary colorectal carcinomas. These results indicate that inactivation of the MTS1 gene is not involved in pathogenesis of primary colorectal carcinomas, and that its enhanced expression is, instead, associated with primary colorectal carcinomas, especially markedly in the early stage of tumor progression.

Antimetastatic effect of PSK, a protein-bound polysaccharide, against the B16-BL6 mouse melanoma.

We examined the effect of PSK, a protein-bound polysaccharide, upon in vivo metastasis and in vitro invasion of the B16-BL6 mouse melanoma cells. (1) PSK suppressed in vivo artificial and spontaneous lung metastases of B16-BL6 in C57BL/6 mice. (2) PSK in a dose-dependent fashion suppressed in vitro invasion and chemotaxis of the tumor cells using filters coated with a reconstituted basement membrane. (3) PSK had little effect on DNA synthesis in tumor cells in vitro, but suppressed tumor cell adhesion to, degradation of, and haptotaxis to components of the basement membrane. (4) PSK suppressed the binding of tumor cells to components of the basement membrane. These findings suggest that PSK may suppress metastasis through inhibition of tumor cell invasion and that this effect is the result of interactions between PSK and components of the basement membrane.

[Mucosal carcinoma of the esophagus--the pathological point of view by subclassification of depth of invasion].

We studied 58 cases of mucosal carcinoma of the esophagus and 21 cases of submucosal carcinoma to evaluate the macroscopic features and histological risk factors (lymphatic and venous permeations and lymph node metastasis) by subclassification of depth of invasion (m1, m2, m3, sm1, sm2, sm3). Carcinoma of m1 and m2 invasion revealed neither venous permeation nor lymph node metastasis. One case of m2 carcinoma (5%) showed lymphatic permeation, 2 cases of m3 carcinomas (20%) showed lymphatic permeation and 3 cases (27%) had lymph node metastasis. Otherwise "sm" carcinoma revealed a high percentage of lymphatic permeation and lymph node metastasis. Also venous permeation were seen in some cases of "sm" carcinoma. The macroscopic appearance of all mucosal carcinomas was 0-II type and most of the 0-iib type carcinomas were "m1" carcinoma. Clinicopathologically it is useful to subclassify mucosal carcinoma as m1, m2, m3, and "m1" and "m2" carcinomas are indications for endoscopic mucosal resection.

Direct PCR of whole blood and hair shafts by microwave treatment.

We report a simple and rapid method for DNA preparation suitable for PCR by microwave irradiation. When mouse whole blood and hair shafts were directly irradiated and subjected to PCR, a unique gene such as the retinoblastoma susceptibility gene was reproducibly amplified. This method was useful for screening a transgene sequence in transgenic mice.

Antitumor effect of biological response modifier, PSK, on C57BL/6 mice with syngeneic melanoma B16 and its mode of action.

The effects of PSK, a protein-bound polysaccharide, on the survival period and effector cell activity were examined using C57BL/6 mice with melanoma B16. PSK prolonged the survival period of the mice with tumors in a schedule- and dose-dependent manner. However, no life-prolonging effect was observed when carrageenan-treated mice or congenitally athymic mice were used as hosts. PSK enhanced the cytostatic activity and interleukin-1-producing capacity of peritoneal exudate plastic-adherent cells in C57BL/6 mice with tumors. These findings suggested that PSK prolongs the survival period of mice with B16 tumors through T-cell- and macrophage-dependent mechanisms.

[Identification of two different genetic mutation associated with silent phenotypes for human serum cholinesterase in Japanese].

Two different gene mutations associated with the silent phenotype for human serum cholinesterase were demonstrated. DNA from five individuals with silent gene phenotype of three unrelated Japanese families was amplified by the polymerase chain reaction (PCR) and analyzed by direct sequencing. The first instance demonstrated a G----C transversion at codon 365 from GGA (Gly) to CGA (Arg), which was seen in three individuals of the two families. This mutation was resulted to create a new Taq 1 restriction site (TCGA). The second mutation was shown by a double heterozygous condition with two different silent gene mutations in two members of remaining one family. These mutations were as follows: 1) one type was a frameshift mutation, in which an extra A was inserted in codon 315 (ACC----AACC) to create a new stop codon at position 322 and 2) the other was the same point mutation at codon 365 as seen in the first instance. These results indicated that many silent variants can be distinguished by direct sequence analyses of genomic DNA.

[Endoscopic needle bladder neck suspension for female stress incontinence].

Thirteen operations of endoscopic bladder neck suspension (Raz method) were performed for 12 patients with stress incontinence between October, 1986 and August, 1988. In all patients, except for one with small bladder capacity, the operations were successful. Disappearance of incontinence was maintained in 10 patients 5-27 months (mean 15.3 months) after the operation, and the success rate of this operation was 77% (10/13) at this follow up point. The operative technique was easy except for the adjustment of pulling up strength of the thread for suspension. We believe that the endoscopic needle bladder neck suspension will be performed more widely because this operation can be done safely in a short time without major complications and can produce very good results.

[Treatment of advanced cancer of the urinary tract with interleukin-2 (IL-2)].

Eight patients with advanced cancer of the urinary tract were treated with recombinant interleukin-2(IL-2) at our department between December, 1985 and September, 1987. Patients were given 5 x 10(5)-4 x 10(6) units of IL-2 by intravenous drip infusion daily. One of five patients who received IL-2 for over 28 days showed 62% regression in size of hepatic metastasis of bladder cancer on computed tomography and another showed marked improvement of Performance Status. The upward tendency of natural killer and lymphokine-activated killer activity of peripheral blood was observed during treatment. Fever, mental disturbance, hypotension and eosinophilia and others were recognized during administration of IL-2, and cardiac failure and disturbance of renal function were also recognized as severe side effect.

Fate and distribution of an antitumor protein-bound polysaccharide PSK (Krestin).

The fate of 14C-labelled PSK in the body was investigated. Although only substances with low mol. wt were observed in blood shortly after the administration, with time, substances with high mol. wt appeared, suggesting absorption of PSK in its original form from the digestive tract. 14C-labelled PSK was distributed in bone marrow, salivary gland, brain liver, spleen, pancreas and tumor. Approximately 70% of 14C-labelled PSK was excreted by expiratory air after 24 h and approximately 15-20% in urine after 72 h. Only a small amount of 14C-labelled PSK was transferred into lymph and bile. The present study on the in vivo behavior of PSK provides an important basis for further analysis of its pharmacological actions.

[Combined therapy of bromocriptine, clomiphene citrate and HMG in normoprolactinemic amenorrhea patients who fail to respond to bromocriptine-clomiphene citrate therapy].

The purpose of this study is to investigate the ovulatory effect of the following three methods on 40 patients with normoprolactinemic amenorrhea patients who failed to respond to clomiphene citrate (Cl) alone. Method I is the combined therapy with bromocriptine and Cl (Brc/Cl). Method II is the combined therapy with Brc/Cl and an additional small amount of HMG (less than or equal to 300 IU). Method III is the combined therapy with Brc/Cl and an increased amount of HMG (less than or equal to 1,200 IU) and HCG. The ovulation rate of method I was 57.5% (23/40) in 40 cases, and 55.6% (99/178) in 178 cycles. The pregnancy rate for this method was 23.8% (5/21). Out of the 12 nonrespondent method I cases, 5 additional cases were ovulated by method II and 3 cases were ovulated by method III. The total pregnancy rate was 42.9% (9/21) with these three methods. Seven cases were normal pregnancies, one case was a twin pregnancy and 1 case was an abortion. No other side effects were found in this study. From these results, the combined therapy with Brc/Cl or Brc/Cl + HMG had almost the same ovulatory rate as the HMG-HCG therapy, and fewer side effects.

[The combined effect of bromocriptine and clomiphene citrate on normoprolactinemic anovulation; the lack of effectiveness of clomiphene citrate treatment alone].

The purpose of this study is to investigate the combined effect of bromocriptine and clomiphene citrate (Cl) treatment on 35 patients with normoprolactinemic amenorrhea who failed to respond to Cl alone. Ovulation was restored by this treatment in 20 of these patients (57.1%), and the pregnancy rate was 26.7%. This treatment was effective in 11 of 15 women with polycystic ovary like syndrome (73.3%). Resting levels of prolactin, estradiol and testosterone in the effective group were significantly higher than those in the non-effective group. As a result of the treatment, the prolactin and LH levels were significantly decreased, and estradiol and progesterone levels were significantly increased in the effective group. Before the treatment, the responsiveness of LH to LHRH in the effective group was significantly higher, but the responsiveness of prolactin to TRH in this group was not significantly higher than that in the non-effective group. After the treatment, the LH-releasing response following a premarin injection in the effective group was significantly higher than that in the non-effective group. The present results indicate that the therapeutic effect of this treatment may be primarily due to the restoration and improvement of the impaired hypothalamo-pituitary system without the suppression of pituitary prolactin secretion.

[Severe cyclophosphamide hemorrhagic cystitis controlled with transurethral electrocoagulation: a case report].

A case of massive cyclophosphamide hemorrhagic cystitis is presented. A 47-year-old woman with Evans syndrome was treated with a total dose of 56.6 g of cyclophosphamide for 2 years. Six months after the cessation of administration, massive hemorrhagic cystitis suddenly occurred following a common cold. Embolization of bilateral internal iliac arteries was ineffective. Therefore we performed transurethral electrocoagulation broadly for bleeding mucosa. The day after operation, bleeding from the bladder remarkably decreased and further transfusions were hardly required.

[Treatment of advanced prostatic carcinoma with cis-diamminedichloroplatinum].

Chemotherapeutic effects of CDDP used as the main drug were studied in 20 patients with progressive prostatic cancer in stage C or D. On the average 208 mg of CDDP was given to the patients receiving chemotherapy without antiandrogen therapy (13 patients who showed resistance to hormone and an untreated new patient) and both ADM and IFM were also given to 3 of them. According to the criterion proposed by Shida and his coworkers, the chemotherapy without antiandrogen therapy was effective in 2 cases, relatively effective in 7 cases, and ineffective in 5 cases. The chemotherapy was effective for metastatic tumors of the lung in 2 out of 2 cases, but had no effect on tumors of the lymph node (1 case) and primary lesion of the tumors (14 cases). The chemotherapy improved acid phosphatase values in 5 out of 10 cases, alkaline phosphatase values in 3 out of 10 cases, dysuria in 4 out of 8 cases, nocturia in 1 out of 12 cases, residual urine in 5 out of 6 cases, lumbago in 6 out of 8 cases, and constitutional symptom in 6 out of 12 cases. The effect of the chemotherapy in combination with antiandrogen therapy was excellent in 4 and good in 2 of the 6 patients treated with castration + diethylstilbestrol diphosphate + CDDP + ADM +/- IFM. The chemotherapy with antiandrogen therapy had no effect on metastatic tumors of the bone (2 cases), but decreased the hardness and size of primary lesion in 6 out of 6 cases. Urethrography showed better changes in 6 out of 6 cases.(ABSTRACT TRUNCATED AT 250 WORDS)

[Chemotherapy of metastatic renal cell carcinoma with cis-diamminedichloroplatinum and other drugs].

Seven patients were entered in this study between November 1978 and February 1983. All patients had histologically proven renal cell carcinoma with widespread metastases. Patients ranged in age from 27 to 69 years, with an average of 54 years. All those studied had measurable radiological lesions of malignant disease. One patient was treated with only CDDP and the other 6 patients were treated with CDDP and other anti-cancer drugs (vinblastine, ifosfamide, bleomycin and/or adriamycin). Total doses of CDDP ranged from 20 to 250 mg, with an average of 99 mg. Four cases showed no change and 3 patients had progressive disease. Symptomatic improvement was obtained in only 2 patients. Management of metastatic renal cell carcinoma is discussed briefly.

[Chemotherapy of disseminated germ cell testicular tumors with cis-diamminedichloroplatinum and other drugs].

The effect of single CDDP therapy and PVB therapy was examined in 7 cases of stage III germ cell testicular tumors with measurable metastases. The mean age of the patients was 30.6 years old, and their histological types of primary sites were seminoma in 3 cases, embryonal carcinoma in 2, immature teratoma in 1 case and embryonal carcinoma + teratoma in 1 case. In 1 case of seminoma, 375 mg of CDDP was administered. In 1 case of embryonal carcinoma + teratoma, 100 mg of CDDP and then 2 courses of PVB therapy were performed, and 3 courses of PVB therapy were given in all other cases. Three cases showed complete response, 2 cases partial response and 2 cases no change. Pulmonary metastatic nodules were extirpated after the PVB therapy in 1 of the cases showing no change, and the histological examination of these nodules was found to be mature teratoma. As a result, the effectiveness of the chemotherapy alone was 71.4%, and that of chemotherapy + surgical operation was 85.7%. Significance of intensive chemotherapy and necessity of extirpation of residual metastatic nodules after intensive chemotherapy in the management of advanced germ cell testicular tumors are stressed.

[Case of immature teratoma of the testicle metastasizing as completely mature teratoma].

A case of immature teratoma of the testicle metastasizing as completely mature teratoma is presented. A 23-year-old man underwent right inguinal orchiectomy for an immature teratoma of the testicle on September 14, 1982. At retroperitoneal lymph node dissection, 12 lymph nodes were removed, all of which were negative for cancer. He was well until March 1983, when bloody sputum and left chest pain occurred. Since full lung tomography revealed two pulmonary nodules, he was treated with a course of VP-16 and three courses of vinblastine, bleomycin and CDDP. In September 1983, after completion of the combination chemo-therapy, the two pulmonary nodules were noticed to be slightly enlarging. A thoracotomy was scheduled to remove these nodules, since they were believed to be his only remaining tumors. Pathologic examination of the extirpated nodules disclosed completely benign teratoma. Mediastinal lymph nodes had no metastatic involvement histologically. It is our intention in this paper to emphasize, by means of several case reports, the frequently benign nature of these residual lesions and also to emphasize a recently recognized phenomenon--the apparent induction of benign teratomas by this intensive chemotherapy.

[Clinical evaluation of transcutaneous oxygen tension measurement for lower limb ischemia].

Transcutaneous oxygen tension (tcPO2) of superficial layer of the foot at 44 degrees C was studied preoperatively in 33 patients (38 legs) with arteriosclerosis (ASO) or thromboangiitis obliterans (TAO). The cases were divided into two groups. The first group consisted of 3 legs, amputated. The second group consisted of 35 legs, not amputated but receiving such treatments as arterial bypass, lumbar sympathectomy or conservative therapy. The tcPO2 index was calculated by dividing the tcPO2 of the chest wall by the tcPO2 of the foot. The value of the tcPO2 of the chest decreased in proportion to the advance of age. There was no significant differences in the tcPO2 indices between ASO and TAO, nor among different sites of peripheral vascular occlusion. On the other hand, as the grade of Fontaine classification increased, the tcPO2 index reduced, with significant difference between grade II and IV. The tcPO2 index of the amputated group was 0.08 +/- 0.04. In contrast, the tcPO2 indices of the lumbar sympathectomy, arterial bypass and conservative therapy groups were 0.74 +/- 0.18, 0.84 +/- 0.12 and 0.74 +/- 0.13, respectively. There was significant difference between the amputated and non-amputated groups. From our results, it is suggested that oxygen tension measured transcutaneously is a useful indicator for the decision of operative indication and surgical procedure.