Do biologics-naïve patients with rheumatoid arthritis respond better to tocilizumab than patients for whom anti-TNF agents have failed? A retrospective study.

OBJECTIVES: To determine responses to tocilizumab between patients with rheumatoid arthritis (RA) who switched to anti-TNF agents and those who are biologics-naïve. METHODS: This retrospective study investigated 107 patients with RA who were treated with tocilizumab. At baseline, 61 of them had already been treated with anti-TNF agents (switched group; 46 for inefficacy and 15 for adverse events), and 46 were biologics-naïve (naïve group). Treatment responses to tocilizumab at week 12 and 24 were compared between the switched and naïve groups using the disease activity score 28 (DAS28). RESULTS: Forty-two (91.3%) and 50 (82.0%) patients in the naïve and switched groups, respectively, completed 24 weeks of tocilizumab treatment. The DAS28-ESR and DAS28-CRP values (means±SD) at weeks 12 and 24 compared to baseline decreased significantly for the naïve and switched groups. The DAS28-ESR and DAS28-CRP values at weeks 12 and 24 were significantly decreased in the naïve group, compared to the switched group. Disease activity was improved in the naïve patients compared to the switched patients. CONCLUSIONS: Tocilizumab was safe, tolerable, and clinically effective for patients with inadequate responses to anti-TNF therapy and for those who were biologics-naïve, and it was more effective among the latter.

Suppression of allergic reactions by royal jelly in association with the restoration of macrophage function and the improvement of Th1/Th2 cell responses.

We studied the immunomodulatory effects of royal jelly (RJ), the principal food source of the queen honeybee. In this study, suppression of allergic reactions by RJ was investigated in DNP-KLH immunized mice (DNP-KLH mice). Oral administration of RJ (1 g/kg) to DNP-KLH mice significantly decreased the serum levels of antigen-specific Ig E and significantly inhibited DNP-KLH mediated-histamine release from mast cells, resulting in the suppression of immediate hypersensitivity reactions of ear skin. In DNP-KLH mice, IFN-gamma (Th1 cytokine) production from CD4+ T cells was suppressed and IL-4 (Th2 cytokine) production from CD4+ T cells was increased as compared to normal mice. On the other hand, RJ improved the balance of Th1/Th2 cell responses from Th2-dominant to Th1-dominant. RJ significantly increased GSH levels in macrophages from DNP-KLH mice. In addition, the administration of RJ to DNP-KLH mice increased IL-12 p40 mRNA expression and NO production, and decreased PG E2 production from macrophages as compared to untreated DNP-KLH mice. These results suggested that RJ suppressed antigen-specific Ig E production and histamine release from mast cells in association with the restoration of macrophage function and improvement of Th1/Th2 cell responses in DNP-KLH mice.

Protease inhibitor reduces loss of tensile strength in rat anastomosis with peritonitis.

BACKGROUND: The tensile strength in intestinal anastomoses decreases postoperatively in association with degradation of the extracellular matrix, and these changes would be expected to be more intense in the presence of peritonitis. MATERIALS AND METHODS: In this study, we investigated extracellular matrix degradation and tensile strength in a rat model of intestinal anastomosis with peritonitis. In the chemical peritonitis model, peritonitis was induced 24 h earlier with intraperitoneal HCl. A serine protease inhibitor, nafamostat mesilate (NM), was given intraperitoneally to some animals every 12 h from immediately after the operation for 3 days. Immunostaining was performed by the standard streptavidin-biotin-peroxidase method after fibronectin (Fn) and factor XIII antigen retrieval on paraformaldehyde-fixed, paraffin-embedded tissue sections. RESULTS: In comparison with controls, administration of NM reduced the loss of tensile strength on Day 3 in a dose-dependent manner, and high-dose NM (20/mg/kg) significantly prevented the loss of tensile strength on Day 3 (P < 0. 05). In the control group, degradation of the collagen layer in the anastomosis was associated with disappearance of Fn and factor XIII staining on Day 3. The administration of NM attenuated these changes with intense immunostaining for Fn and factor XIII seen particularly between collagen fibers on both sides of the anastomosis on Day 3. In the chemical peritonitis model, administration of NM also significantly prevented the loss of tensile strength on Day 3 without disappearance of collagen fibers. CONCLUSION: These findings suggest that NM may be clinically useful for preventing intestinal leakage, particularly when anastomoses are performed under protease-activating conditions, such as intestinal edema and inflammation.

Differential distribution of TRP Ca2+ channel isoforms in mouse brain.

Mammalian homologues of the Drosophila TRP proteins, which are essential for light-activated, phosphatidyl-inositide (PI)-dependent Ca2+ conductance in Drosophila photoreceptors, were molecularly identified, to investigate receptor-activated Ca2+ influx in the mammalian nervous system. Two cloned mouse TRP homologues, TRP3 and TRP4, structurally related to the voltage-dependent Na+ channel, were expressed predominantly in the brain, where a sharp contrast in the distribution of the RNA transcripts for TRP isoforms was demonstrated by in situ hybridization analysis. TRP3 mRNA was concentrated in cerebellar Purkinje cells and sparsely localized in the cerebellar granule layer, pontine nuclei, and thalamus, whereas TRP4 mRNA was abundantly expressed in hippocampal CA1 pyramidal neurons, dentate gyrus granule cells, and cerebral cortical neurons, and in the septal nuclei and the mitral layer of olfactory bulb. The distinct spatial patterns of TRP isoforms implicate that neurons are highly heterogeneous in receptor-activated Ca2+ influx responsible for the second phase of PI-mediated rise in intracellular Ca2+.

Methotrexate maintains bone mass by preventing both a decrease in bone formation and an increase in bone resorption in adjuvant-induced arthritic rats.

We examined the effects of low doses methotrexate (MTX) and indomethacin (IND) on bone mass and turnover in normal male Sprague-Dawley rats and those with adjuvant-induced arthritis. Normal and the adjuvant (heat-killed mycobacterium)-injected rats, 6 weeks of age, were given MTX at daily doses of 0.05, 0.1, or 0.2 mg/kg body weight (BW) or IND at a daily dose of 1.0 mg/kg BW. Rats were killed at the start, or at 14 and 28 days. In normal rats, the administration of these agents did not change the lumbar and femoral BMD values, nor did the serum osteocalcin or urinary deoxypyridinoline (D-Pyr) levels. Lumbar trabecular osteoclast number (Oc.N/BS) and osteoclast surface (Oc.S/BS) were decreased in the rats given IND. In the arthritic rats, the administration of MTX did not prevent an early increase of paw edema in the adjuvant-injected limb, but late inflammatory edema was alleviated in the non-injected limb. However, MTX administration at a dose of 0.1-0.2 mg/kg BW maintained an age-dependent increase in the lumbar and femoral BMD values. While serum osteocalcin levels were decreased and urinary D-Pyr values were increased in the arthritic control rats, these bone markers remained at the levels of the normal rats. Decreases in mineral apposition rate (MAR) and bone formation rate (BFR/BS) and increases in the trabecular Oc.N/BS and Oc.S/BS values were prevented by MTX. While IND almost completely prevented inflammatory paw edema, it did not improve the parameters of bone formation. An increase in osteoclasts was prevented and the osteopenia in the lumbar and the femoral bone was only partially prevented by IND. These data suggest that MTX improves bone mass and turnover in the arthritic rat, in which several cytokines that affect bone cells are involved. An increase in bone resorption may be due to prostaglandins, but bone formation defect was suggested to be due to other cytokines such as IL-1, IL-6, and TNF-alpha in this model.

Measurement of 5-fluorouracil in scalp hair: a possible index of patient compliance with oral adjuvant chemotherapy.

OBJECTIVE: Little is known about patient compliance with oral adjuvant chemotherapy. It is estimated to be poor especially in Japan, where it is still unusual for patients to be directly informed of their diagnosis of malignancy. 5-Fluorouracil (5-FU) was measured in hair samples to assess patient exposure to 5-FU, and its potential usefulness is discussed as an index of compliance with postoperative adjuvant chemotherapy. METHODS: Hair samples obtained from 55 patients, who had received oral 5-FU (total dose 27-41 g) as postoperative adjuvant chemotherapy over a 6-month period, were used for the analysis of 5-FU. The drug was extracted from the hair using ethyl acetate, and its fluorescence derivatization was employed for measurement with HPLC. The detection limit of 5-FU in hair was 0.01 ppm. RESULTS: In 22 out of 55 samples 5-FU content was under the detection limit, whereas in the remaining 33 samples the drug was detected in a range of 0.006-2.125 ng per hair strand; in addition, drug content showed a lognormal distribution. 5-FU was detected in the hair collected from those patients who were possibly compliant with the postoperative oral adjuvant chemotherapy. CONCLUSION: As many as 40% of the patients analysed were supposed to be much less compliant. Even in the possibly compliant patients, the degree of compliance with the therapy varied according to a log-normal distribution.

Prospective study of chemoprevention of hepatocellular carcinoma with Sho-saiko-to (TJ-9).

BACKGROUND: Most hepatocellular carcinomas (HCC) arise in patients with cirrhosis, in whom its incidence is high. The prevention of HCC in patients with cirrhosis is important. METHODS: A prospective, randomized, nonblind controlled study was performed to evaluate the preventive effect of Sho-saiko-to (TJ-9) on HCC development. TJ-9 is a Chinese herbal medicine that contains crude extracts of seven herbs; it has antitumor effects in experimental animals. Two hundred sixty patients with cirrhosis were randomly assigned to two groups, matched for age, sex, presence of hepatitis B surface antigen, and the severity of liver damage. The patients in the trial group were given TJ-9 at a daily oral dose of 7.5 g in addition to the conventional drugs given to the control patients. The patients were prospectively monitored for 60 months and the cumulative incidence of HCC and the survival rate in the two groups were calculated. RESULTS: The cumulative incidence curve for 5 years of the trial group was lower than that of the control group (P = 0.071). For the patients without HBs antigen, the difference was significant (P = 0.024). The survival curve for 5 years of the trial group was higher than that of the control group (P = 0.053). For the patients without HBs antigen, the difference was significant (P = 0.043). CONCLUSIONS: TJ-9 helped to prevent the development of HCC in patients with cirrhosis, particularly in patients without HBs antigen.

Characterization of mitogenic substances in the hot water extracts of bupleuri radix.

Bupleuri Radix is a commonly used medicinal plant in Kampo medicine, and its hot water extracts show mitogenic activity to murine lymphocytes. In this paper the mitogenic substances in the hot water extracts of Bupleuri Radix (Bup-HWE) were fractionated and characterized physicochemically and immunologically. Most of these substances were recovered from mol. wt of more than 200 kDA fraction (fr. C-13). Separation of fr. C-13 by phenol-water fractionation method gave water soluble and phenol soluble mitogenic substances. These substances showed the activity even in C3H/HeJ mice, and polymyxin B or lysozyme treatment did not abrogate the activity, suggesting that the active substances are not related to bacterial lipopolysaccharide. Treatment of the mitogenic substances recovered from the phenol layer with NaCLO2, a polyphenol degrading chemical, significantly reduced the activity, but pronase and pectinase treatments were not effective. The mitogenic substances in the water layer were active even after NaCLO2 treatment. These findings suggested that the mitogenic substances of Bup-HWE are large molecular weight polyphenolic compounds and polysaccharide. The mitogenic substances are suggested to be B cell mitogens.

[A case of adult onset hypophosphatemic vitamin D resistant osteomalacia].

A case of adult onset hypophosphatemic vitamin D resistant osteomalacia is described. A 40-year-old female who complained of thorax and lumbar pain and gait disturbance was admitted to our hospital on 7 November, 1988. The patient had hypophosphatemia with normal plasma calcium, parathyroid hormone and 25-hydroxy vitamin D3 concentrations, but had decreased tubular reabsorption of phosphate and decreased plasma 1, 25-dihydroxy vitamin D3 concentrations. The iliac crest bone biopsy showed osteomalacic changes. The 99mTc-MDP bone scintigram showed evidence of increased bone turnover with raised plasma alkaline phosphatase concentrations. After treatment with oral 1 alpha-hydroxy vitamin D3 (3-6 micrograms/day) and intravenous or oral phosphate supplement (0.47-1.74g/day), the subjective and clinical findings improved.

Morphological features of cat thalamo-parietal projection fibers investigated by PHA-L immunohistochemistry and electron microscopy.

Thalamo-parietal fibers originating from the ventroanterior-ventrolateral (VA-VL) complex in the cat were labeled with Phaseolus vulgaris leucoagglutinin (PHA-L) and examined by light and electron microscopy. PHA-L (2.5% aqueous solution) was injected iontophoretically through micropipets with anodal current pulses into the VA-VL complex. PHA-L-labeled terminals were distributed in the lateral and the suprasylvian gyri in the superficial and deep cortical layers. In layer I, horizontal varicose fibers and terminals were conspicuous in the upper one-third and were widely distributed. In the deeper cortical layers (layers III-V), varicose fibers and terminals were detected in moderate numbers. Electron microscopic examination revealed that the labeled terminals formed asymmetrical synapses on the dendritic spines of spiny neurons. These morphological findings appeared to be consistent with our previous intracellular recordings in this cortex.

Randomized, controlled study on adjuvant immunochemotherapy with PSK in curatively resected colorectal cancer. The Cooperative Study Group of Surgical Adjuvant Immunochemotherapy for Cancer of Colon and Rectum (Kanagawa).

A randomized, controlled trial of adjuvant immunochemotherapy with PSK (Kureha Chemical Industry Co., Tokyo, Japan) in curatively resected colorectal cancer was studied in 35 institutions in the Kanagawa prefecture. From March 1985 to February 1987, 462 patients were registered. Four hundred forty-eight of those patients (97.0 percent) satisfied the eligibility criteria. The control group received mitomycin C intravenously on the day of and the day after surgery, followed by oral 5-fluorouracil (5-FU) administration for over six months. The PSK group received PSK orally for over three years, in addition to mitomycin C and 5-FU as in the control group. At the end of February 1990, the median follow-up time for this study was four years (range, three to five years). The disease-free survival curve and the survival curve of the PSK group were better than those of the control group, and differences between the two groups were statistically significant (disease-free survival, P = 0.013; survival, P = 0.013). These results indicate that adjuvant immunochemotherapy with PSK was beneficial for curatively resected colorectal cancer.

Intracellular recordings from rat thalamic VL neurons: a study combined with intracellular staining.

Intracellular recordings from thalamic neurons receiving cerebellar inputs were performed under urethane anesthesia in the rat. A total of 64 neurons were recorded intracellularly with micropipettes filled with 4% biocytin solution (dissolved in 0.5 M K-acetate), and cerebellar-induced EPSPs (CN-EPSPs), the membrane resistance and firing properties were analyzed with intracellular current injections. The mean latency of CN-EPSPs was 1.9 +/- 0.8 ms and the mean input resistance measured in 10 neurons was 17.6 +/- 5.0 M omega. Thirty-two out of 35 stained neurons were analysed morphologically; 28 of these neurons were located in the VL, and 26 received CN-EPSPs. Their somata were round or polygonal in shape and the mean size was 22.5 x 15.2 microns. They had radially extending spinous dendrites, and the mean radii of the dendritic fields were 214.7 microns in the frontal and 171.4 microns in the sagittal planes. These morphological features were similar to those observed in the sensory relay nucleus of the thalamus.

Cortical laminar distribution of rat thalamic ventrolateral fibers demonstrated by the PHA-L anterograde labeling method.

The cortical innervation pattern of rat thalamic ventrolateral (VL) fibers was investigated with the Phaseolus vulgaris-leukoagglutinin (PHA-L) anterograde labeling method. The PHA-L-labeled terminals were distributed in the sensorimotor cortex where the cerebello-cerebral (C-C) evoked responses were recognized. The labeled terminals were detected in all layers except layer VI. In the upper part of layer I, the most densely packed terminals were formed and moderately labeled terminals were seen in layers IV and V. Such a laminar distribution of PHA-L-labeled VL terminals is consistent with the profiles of the C-C responses according to laminar field potential analysis.

Improved high-speed counter-current chromatograph with three multilayer coils connected in series. II. Separation of various biological samples with a semi-preparative column.

The semipreparative capability of the newly developed high-speed counter-current chromatograph equipped with a set of three multilayer coils has been demonstrated in separations of a variety of biological samples including triterpenoic acids, indole auxins, bacitracin, flavonoids and tetracycline derivatives, each with a suitable two-phase solvent system. The sample quantities ranging from 50 to 500 mg were efficiently separated within a few hours. The separation of tetracycline derivatives was remarkably improved by adding ammonium acetate to the solvent system.

[Randomized controlled study on adjuvant immunochemotherapy with PSK in curatively resected colorectal cancer. The Cooperative Study Group of Surgical Adjuvant Immunochemotherapy for Cancer of Colon and Rectum].

To evaluate of adjuvant immunochemotherapy with PSK in curatively resected colorectal cancer, randomized controlled study by 35 institutions in Kanagawa prefecture was conducted. From March 1985 till February 1987, 462 patients were assigned one of two different regimens. 448 patients (97.0%) of them satisfied the eligibility criteria. Control group received mitomycin C intravenously on the day and the day after the operations respectively followed by 5-FU orally over for 6 months. PSK group received in addition to mitomycin C and 5-FU as in control group, PSK orally for over 3 years. By February 1989, follow up studies of the patients after their operations had been carried out for two years to four years. The disease free curve and the survival curve of PSK group were higher than those of control group, differences between the two groups were statistically significant (Disease free curve: P = 0.0096, survival curve: p = 0.0391). From these results, adjuvant immunochemotherapy with PSK was considered beneficial for curatively resected colorectal cancer.

[Controlled prospective trial to evaluate Syosakiko-to in preventing hepatocellular carcinoma in patients with cirrhosis of the liver].

A controlled prospective study was conducted to evaluate the potential of Syo-saiko-to (Xiao-Chai-Hu-Tang) for the prevention of hepatocellular carcinoma (HCC). Pairs of patients were matched for age, sex, presence of HBs antigen and the scores of the severity of liver dysfunction from 260 cirrhotic subjects. We randomly assigned each patient to receive either a conventional medicine (control group), or 7.5 g/day of Syo-saiko-to (trial group). The patients were monitored during 34 months of treatment, and the incidence of HCC in the two groups were compared. Seventeen patients were found to have HCC in the control group, and nine were found to have HCC in the trial group. The incidence of HCC was significantly lower in the trial group. The results of this study suggested that Syo-saiko-to may prevent or delay the emergence of latent HCC in patients with cirrhosis of the liver.

Treatment of chronic liver injury in mice by oral administration of xiao-chai-hu-tang.

Oral administration of Xiao-Chai-Hu-Tang, the extract of a mixture of seven herbs, attenuated the hepatic fibrosis developed in mice after repeated doses of carbon tetrachloride. Its pre-administration reduced the derangement of liver function tests seen after a single dose of carbon tetrachloride as well as that seen after d-galactosamine intoxication. Xiao-Chai-Hu-Tang may be effective in the treatment of chronic liver injury through hepatocytoprotection.