The study on the immunological effect of sizofilan combined with radiotherapy in patients with uterine cervical cancer

To investigate the immunological effect of Sizofilan (SPG) combined with radiotherapy, we evaluated the immunological parameters in 22 patients with uterine cervical cancers. Twelve cases were treated with SPG combined with radiotherapy (SPG group), and the other ten cases, with radiotherapy only (control group). As a result, 1) During radiotherapy, the numbers of lymphocyte and CD2 positive cell decreased in SPG and control groups. After radiotherapy, however, its numbers in SPG group became significantly higher than in control group (p less than 0.05). The number of CD3 positive cell also presented a tendency to increase after radiotherapy in SPG group. As for CD20 positive cell, its numbers were kept unchanged after radiotherapy in both two groups, and no significant difference was observed between them. 2) NK cell activity decreased during radiotherapy in both two groups. After radiotherapy, its activity in SPG group recovered to its pre-value and became significantly higher than that in control group (p less than 0.05). 3) SPG did not any prominent effect on CD4/CD8 ratio. 4) The adverse effect of SPG to liver or kidney function were not observed in our patients. The SCC level in SPG group decreased rapidly by radiotherapy as well as that in control group, and no significant difference was observed in SCC levels between them. So it was suggested that SPG did not suppress the cytocidal effect of radiation to cancer cells. Based on these findings, it was concluded that SPG prompted the recovery of not only lymphocyte, especially T cell, but also NK cell activity. These immunological findings presented a usefulness of clinical application of SPG to radiotherapy in patients with uterine cervical cancers.     

Behavioral and neuroanatomical analyses in a genetic mouse model of 2q13 duplication

Duplications of human chromosome 2q13 have been reported in patients with neurodevelopmental disorder including autism spectrum disorder. Nephronophthisis‐1 (NPHP1) was identified as a causative gene in the minimal deletion on chromosome 2q13 for familial juvenile type 1 nephronophthisis and Joubert syndrome, an autosomal recessive neurodevelopmental disorder characterized by a cerebellar and brain stem malformation, hypotonia, developmental delay, ataxia, and sometimes associated with cognitive impairment. NPHP1 encodes a ciliary protein, nephrocystin‐1, which is expressed in the brain, yet its function in the brain remains largely unknown. In this study, we generated bacterial artificial chromosome‐based transgenic mice, called 2q13 dup, that recapitulate human chromosome 2q13 duplication and contain one extra copy of the Nphp1 transgene. To analyze any behavioral alterations in 2q13 dup mice, we conducted a battery of behavioral tests. Although 2q13 dup mice show no significant differences in social behavior, they show deficits in spontaneous alternation behavior and fear memory. We also carried out magnetic resonance imaging to confirm whether copy number gain in this locus affects the neuroanatomy. There was a trend toward a decrease in the cerebellar paraflocculus of 2q13 dup mice. This is the first report of a genetic mouse model for human 2q13 duplication.     

A longitudinal study on the prognosis of ovulatory disturbance in teenage patients with high LH and normal FSH serum levels

To study the prognosis of adolescent ovulatory disturbance in patients with persistently elevated LH levels (greater than or equal to 25 mIU/ml), normal FSH levels and high LH/FSH (greater than 2.0), 17 patients aged 12-19 years were studied longitudinally for 4-9 years. These 17 patients consisted of 7 patients suffering from amenorrhea with estrogenic effect, 5 patients with functional bleeding, 3 patients with delayed menarche and 2 patients with oligomenorrhea. All of the patients showed exaggerated LH responses to 100 micrograms of LHRH administration while the FSH responses were not different from those obtained from normal women. Out of the 17 patients, 10 (58.8%) patients showed the values of testosterone and 7 (41.2%) androstenedione which were above the mean + 2SD of normal women. Consequently, the mean serum testosterone and androstenedione levels were significantly higher than those in normal women. The mean LH (36.6 +/- 8.3 mIU/ml), FSH (11.2 +/- 1.5 mIU/ml) and LH/FSH (3.3 +/- 0.8) at the age of 21.4 +/- 2.5 years were not different from the mean LH (39.9 +/- 13.3 mIU/ml), FSH (10.8 +/- 1.8 mIU/ml) and LH/FSH (3.8 +/- 1.5) at the age of 16.1 +/- 1.8 years, respectively. None of the 17 patients showed amelioration or deterioration of ovulatory disturbance during long-term observation. To further investigate the central dopamine activity, 10 mg of metoclopramide (MCP) was administered intravenously in these 17 patients. The LH and PRL responses to MCP were evaluated, and the results were compared to those obtained from 17 patients aged over 20 with PCO and from 17 normal women. The LH responses to MCP were positive in this juvenile patient group and the patients aged over 20 PCO group. However, the LH responses to MCP were negative in normal women in both the follicular and luteal phases. In contrast, the PRL responses to MCP were significantly attenuated in juvenile patients and in patients aged over 20 with PCO compared to those in normal women. Since the hormonal profiles in these 17 patients with anovulation or oligo-ovulation were very similar to those in the group aged over 20 with established PCO, it may be suggested that 1) at least part of the adult patients with PCO may have had PCO from late adolescence; 2) the majority of the patients with high LH and normal FSH levels in adolescence will suffer from ovulatory disturbance continuously; 3) in these patients, an aberration of central dopamine in control of LH and PRL may exist.     

Intermittent chemotherapy can retain the therapeutic potential of anti‐CD137 antibody during the late tumor‐bearing state

Immunomodulating monoclonal antibodies (mAb) can evoke antitumor T‐cell responses, which are attenuated by regulatory T cells (Treg) and myeloid‐derived suppressor cells (MDSC). Treatment with cyclophosphamide (CP) and gemcitabine (GEM) can mitigate the immunosuppression by Treg and MDSC, respectively. In the current study, we examined the antitumor effects of a combination of local injection with anti‐CD137 mAb and intermittent low‐dose chemotherapy using CP and GEM in subcutaneously established CT26 colon carcinoma. Although a significant antitumor effect was observed when local anti‐CD137 mAb therapy (5 μg) was started early in the tumor‐bearing stage (day 10), no therapeutic efficacy was observed when the mAb therapy was started at a later tumor‐bearing stage (day 17). Analyses of the tumor‐infiltrating immune cells revealed that the number of Gr‐1^high/low CD11b⁺ MDSC started to increase 13 days after tumor inoculation, whereas injection with low‐dose (50 mg/kg) CP and GEM mitigated this increase. In addition, although intermittent injections with low‐dose CP and GEM on days 10 and 18 suppressed tumor growth significantly, additional local injections of anti‐CD137 mAb on days 19, 21, and 23 further augmented the therapeutic efficacy. Cytotoxic T lymphocytes reactive to CT26 and a tumor antigen peptide were induced successfully from the spleen cells of tumor‐cured or tumor‐stable mice. In a bilateral tumor inoculation model, this combination therapy achieved systemic therapeutic effects and suppressed the growth of mAb‐untreated tumors. These results suggest that intermittent immunochemotherapy using CP and GEM could retain the therapeutic potential of anti‐CD137 mAb that is normally impaired during the late tumor‐bearing stage.     

Immune checkpoint blockade opens an avenue of cancer immunotherapy with a potent clinical efficacy

Recent progress in tumor immunology has revealed that tumors generate immunologically restrained milieu during the process of their growth, which facilitates the escape of tumors from host immune systems. Immune checkpoint molecules, which transduce co‐inhibitory signals to immuno‐competent cells, are one of the most important components conferring the immunosuppressive capacity in the tumor microenvironment. Cytotoxic T‐lymphocyte‐associated protein 4 (CTLA‐4) and programmed cell death‐1 (PD‐1) are typical immune checkpoint molecules intimately involved in the suppression of anti‐tumor immunity. Antibodies against those molecules have been developed, such as ipilimumab (anti‐CTLA‐4 antibody), nivolumab and pembrolizumab (anti‐PD‐1 antibody), and have been approved by regulatory agencies and used in some countries. Treatment with these antibodies demonstrates previously unobserved clinical efficacies superior to the conventional therapies. In this review, we first discuss the escape mechanisms of cancer from host immune systems, and then focus on the recent advances in immune checkpoint blockade therapy and on the new findings of related immune reactions, aiming to provide a better understanding of the novel cancer immunotherapies.     

Simultaneous changes in high-fat and high-cholesterol diet-induced steatohepatitis and severe fibrosis and those underlying molecular mechanisms in novel SHRSP5/Dmcr rat

Objectives

The aim of this study was to identify the molecular mechanisms underlying high-fat and high-cholesterol (HFC) diet-induced steatohepatitis and associated liver fibrosis progression in a novel stroke-prone, spontaneously hypertensive 5/Dmcr (SHRSP5/Dmcr) rat model.

Methods

SHRSP5/Dmcr rats were given the control or HFC-diet for 2, 8, and 16 weeks. Plasma and hepatic gene expression of key molecules involved in fatty acid oxidation, inflammation, oxidative stress, and fibrosis were subsequently analyzed.

Results

Rats fed the HFC-diet showed increased plasma tumor necrosis factor-α (TNF-α) and hepatic p50/p65 signals, but reduced hepatic Cu²⁺/Zn²⁺-superoxide dismutase across the treatment period and reduced plasma total adiponectin at 8 weeks. In HFC-diet-fed rats, transforming growth factor-β1 (TGF-β1) was elevated prior to the appearance of obvious liver fibrosis pathology at 2 weeks, followed by elevations in platelet-derived growth factor-B (PDGF-B) and α-smooth muscle actin (α-SMA), corresponding to evident liver fibrosis, at 8 weeks and by α₁ type I collagen production at 16 weeks. The HFC-diet increased hepatic total cholesterol accumulation, although hepatic triglyceride declined by 0.3-fold from 2 to 16 weeks due to reduced hepatic triglyceride synthesis, as suggested by the diacylglycerol acyltransferase 1 and 2 measurements.

Conclusions

TNF-α and p50/p65 molecular signals appeared to be major factors for HFC-diet-induced hepatic inflammation and oxidative stress facilitating liver disease progression. While the up-regulation of TGF-β1 prior to the appearance of any evident liver fibrosis could be an early signal for progressive liver fibrosis, elevated PDGF-B and α-SMA levels signified evident liver fibrosis at 8 weeks, and subsequent increased α₁ type I collagen production and reduced triglyceride synthesis indicated extensive liver fibrosis at 16 weeks in this novel SHRSP5/Dmcr model.

Electronic supplementary material

The online version of this article (doi:10.1007/s12199-012-0273-y) contains supplementary material, which is available to authorized users.     

Experimental gastric carcinoma induced by N-methyl-N′-nitro-N-nitrosoguanidine

Summary Serial studies by endoscopy and biopsy were made in a Beagle dog during and after oral administration of N-methyl-N-nitro-N-nitrosoguanidine (MNNG). Between the 23rd and the 45th week of observation erosions and ulcers appeared at the angulus of the stomach and turned into ulcer scar. A depression with atypical glands was seen in the ulcer scar of the posterior wall of the angulus at the 94th week. It developed elevated margins at the 102nd week, when a well differentiated adenocarcinoma was found histopathologically. Ulceration and reepithelialization were observed in the early carcinoma. The carcinoma progressed into a larger one of Borrmann's type 2 at the 115th week and further into its type 3 at the 181st week. A second carcinoma with signet ring cell carcinoma developed in the anterior wall of the angulus. The two carcinomas fused and formed a single lesion. At autopsy in the 216th week the carcinoma invaded the serosa, and metastasis to regional lymph nodes was observed.Supported by grants from the Ministry of Education, Science, and Culture, the Ministry of Health and Welfare, and Nakayama Foundation in Japan     

Changes in polyamine metabolism of rat liver after administration of D-galactosamine. Favorable effects of putrescine administration on galactosamine-induced hepatic injury.

There are many reports showing a close relation between polyamine metabolism and tissue growth or recovery of damaged tissues, such as regenerating liver. Thus, changes in polyamine metabolism in the livers from rats treated with D-galactosamine, an inducer of experimental hepatitis, were studied. The activity of ornithine decarboxylase started to increase 14 hr after administration of galactosamine and reached 30 times the normal activity at about 25 hr, the time of maximum severity of hepatitis. The content of putrescine increased to about 10 times the control value. After increases in the putrescine content and ornithine decarboxylase activity, the hepatitis started to diminish. Increases in the activity of S-adenosylmethionine decarboxylase and the content of spermidine were observed 33-37 hr after administration of galactosamine. The maximum values of these parameters, which were significantly higher than the control values, were observed after the healing process had started.