IL-12p40 gene expression in lung and hilar lymph nodes of MPS-resistant pigs.

Mycoplasma pneumonia of swine (MPS) is caused by Mycoplasma hyopneumoniae (M.hp) and is a common chronic respiratory disease of pigs. Recently, a genetically selected variant of the Landrace pig (Miyagino L2) has a lower incidence of pulmonary MPS lesions. We investigated the pathological and immunological characteristics of MPS resistance in these pigs (n = 24) by comparing with the normal landrace pig (control: n = 24). The pathological MPS lung lesion score in MPS-selected landrace pigs was significantly lower than in the control. The gene expression of interleukin (IL)-12p40, which acts as a chemoattractant and a component of the bioactive cytokines IL-12 and IL-23, was significantly higher at the hilar lymph nodes, lung, and spleen in MPS-selected landrace pigs than in control landrace pigs, and these were negatively correlated with the macroscopic MPS lung lesion score. In summary, we demonstrate that resistance against MPS in Miyagino L2 pigs is associated with IL-12p40 up-regulation, in comparison with normal landrace pigs without the MPS vaccine. In addition, a comparative study of macroscopic MPS lung lesions and IL-12p40 gene expression in lung and hilar lymph nodes may lead to beneficial selection traits for the genetic selection for MPS resistance in pigs.

[Endoscopic Submucosal Dissection(ESD)as a "Loco-Regional Cancer Therapy" for Colorectal Tumors].

INTRODUCTION: Endoscopic submucosal dissection(ESD)for colorectal tumors has been covered by the national health insurance system in Japan since April 2012, and is widely used. We analyzed colorectal ESD cases we performed. PATIENTS AND METHODS: We investigated 515 patients with colorectal lesions(580 lesions)for whom ESD was performed between November 2005 and April 2017. Clinicopathological data, technical methods, complications, and outcomes were analyzed. RESULTS: Most tumors were found in the transverse colon(134 lesions). The average diameter was about 26 mm. The largest lesion was 120 mm. The en bloc resection rate was high(96.2%). The average operative time was 51 minutes. Among complications, the number of delayed major bleeding cases was 7(1.2%). Minor perforations occurred in 3 cases(0.5%). The perforation could be closed with endoscopic clips. About 70% of the cases were adenomas, and the remainder were carcinomas. One patient with carcinoma in situ showed a mucosal recurrence 4 months later and received repeat endoscopic treatment. The cure rate was 99.8%. Among 29 deep submucosal invasion cases, additional colectomy was performed in 21; 3 patients had persistent carcinoma in the colonic wall and another patient had lymph node metastasis. CONCLUSIONS: Colorectal ESD can be performed for all sites in the large intestine, and en bloc resection was possible for a large lesion. A good outcome was observed for "Loco-Regional Cancer Therapy" in early colorectal carcinoma.

Changes in hepato-renal gene expression in microminipigs following a single exposure to a mixture of perfluoroalkyl acids.

It is evident that some perfluoroalkyl acids (PFAAs), a group of globally dispersed pollutants, have long biological half-lives in humans and farm animals. However, the effects of PFAAs in domestic animals have not been fully elucidated. The present study investigated how exposure to a single dose of a mixture of 10 PFAAs influenced hepatic and renal gene expression and histopathology, as well as plasma clinical biochemistry, in microminipigs (MMPigs) over 21 days. In animals treated with PFAAs, the mRNA expression of twelve genes related to fatty acid metabolism was upregulated in the kidney, while only few of these genes were induced in the liver. The expression of several kidney injury-associated genes such as, IGFBP1, IGFBP6, GCLC X2, GCLC X3, MSGT1, OLR1 was upregulated in the kidney. Interestingly, the expression of IGFBP-genes was differentially altered in the liver and kidney. Our findings thus identified hepato-renal gene expression changes in MMPigs that were associated with various molecular pathways including peroxisome proliferation, lipid metabolism, kidney injury, and apoptosis. Furthermore, serum HDL levels were significantly decreased following exposure to PFAAs, whereas no significant histopathological changes were detected, as compared to the vehicle group. Taken together, the present study provided the first indication that a single exposure to a mixture of PFAAs can produce changes in MMPig renal gene expression that were observed three weeks post exposure, suggesting that more attention should be paid to the kidney as a primary target organ of PFAAs.

In vivo effect of a TLR5 SNP (C1205T) on Salmonella enterica serovar Typhimurium infection in weaned, specific pathogen-free Landrace piglets.

Toll-like receptor 5 is a pattern-recognition receptor for bacterial flagellin. We previously reported that a single nucleotide polymorphism (SNP) of swine TLR5, C1205T, impairs recognition of Salmonella typhimurium (ST) flagellin and ethanol-killed Salmonella Choleraesuis (SC). In the present study, weaned, specific pathogen-free (SPF) Landrace piglets with CC, CT or TT genotypes were orally infected with ST (L-3569 strain) to determine the effect of this specific SNP on ST infection in vivo. Eighteen ST-infected piglets (six each with CC, CT, or TT) exhibited fever and diarrhea for 1 week after infection. TT piglets had the longest duration of fever. TT piglets had the greatest mean diarrhea score during the experimental period, followed by CT and CC piglets. Fecal ST shedding was greater in CT and TT pigs than CC pigs from 2 days after infection. Serum haptoglobin concentration increased in ST-infected piglets and to greater extents in CT and TT pigs than CC pigs. Daily weight gain was lower in infected pigs, particularly TT piglets, than control pigs. To the best of our knowledge, this study is the first to demonstrate that impairment of TLR recognition affects pig susceptibility to disease in vivo. Thus, piglets with the T allele of swine TLR5 (C1205T) exhibit impaired resistance to ST infection. Furthermore, elimination of the T allele of this SNP from Landrace pigs would lead to enhancement of their resistance to ST infection.

[Relationship between Surgical Site Infection(SSI)Incidence and Prognosis in Colorectal Cancer Surgery].

We examined the relationship between risk factors for surgical site infections(SSIs)and prognosis in 440 patients who underwent colon cancer surgery. We evaluated factors related to SSIs(GradeB II)and compared survival rates by stage. SSIs were observed in 36 patients. An increased SSI incidence was associated with pulmonary dysfunction, mGPS=2, CONUT≥2, PNI≤40, NLR>4.3, location(rectum), depth of tumor invasion(Bsubserosa, SS), lymph node metastasis, laparotomy, resection of other organs, colostomy, blood loss(large), and operative time(long). Survival rates were lower in Stage II/III patients with SSIs. Preoperative risk judgment is important in colorectal cancer surgery.

[A Case of Recurrence in the Posterior Wall of the Virginal after Radical Resection for Rectal Cancer Well Responded in a Long Period by Chemo-Radiotherapy].

We report a case of an advanced rectal cancer recurrence that responded completely to chemo-radiotherapy. The patient was an 80-year-old woman. Low anterior resection with D2(prxD3)lymph node dissection was performed. Sixteen months after operation, CEA level elevated but no recurrence foci were found in any image tests. Administration of TS-1 was initiated since recurrence was highly suspected. Twenty seven months after operation, PET-CT detected local recurrence in the posterior wall of the vagina. After construction colostomy, chemo-radiotherapy(60 Gy+oral UFT)was performed and CEA level dropped promptly to the normal value. No relapse was pointed out in CT scans or MRI tests. There were not any signs of recurrence through 112 months after chemo-radiotherapy.

Effects of mycoplasmal pneumonia of swine (MPS) lung lesion-selected Landrace pigs on MPS resistance and immune competence in three-way crossbred pigs.

To clarify the genetic influence of mycoplasmal pneumonia of swine (MPS) lesion-selected Landrace (La) on MPS resistance and immune characteristics in three-way crossbred pigs (LaWaDa), the LaWaDa pigs were compared with the non-selected crossbred (LbWbDb) and purebred (La) pigs. The MPS lesion score in the three lines was as follows: La line < LaWaDa line < LbWbDb line, with significant differences among the lines. The proportions of myeloid cells and T cells were lower and higher, respectively, in the LaWaDa pigs compared with those in the other two lines. Messenger RNA (mRNA) expression of interleukin (IL)-6, IL-10, transforming growth factor-ß, and interferon-γ in peripheral blood was significantly increased after vaccination in the La and LaWaDa lines. IL-4 mRNA expression in the LaWaDa line was intermediate to the La and LbWbDb lines. Furthermore, principal component analysis for immune traits and MPS lesions was executed to clarify the characteristics of each pig line. These findings suggest that the immune responses in the three pig lines are genetically distinct and that MPS resistance and some immunity characteristics from the La line were transmitted to the three-way crossbred pigs.

[Prognostic Factors in Elderly Patients with Stage II/III Colorectal Carcinomas with Invasion of pT3 or Deeper].

Three hundred and fourteen patients with pT3 or deeper, Stage II/IIIcolorectal carcinoma who underwent curative operation were divided into two groups: age 75 years and more(elderly)and 74 years and less(non-elderly), to identify prognostic factors and risk factors for postoperative complications. Higher comorbidity rates and poorer nutritional status where found in the elderly group, which led to a shorter operation time and less lymph node dissection. There were no significant differences in 5-year OS and 3-year RFS between the two groups. These results suggest that elderly patients with colorectal carcinoma can achieve outcomes equal to the non-elderly group by curative resection even if it is not standard radical resection.

[A Case of Early Anal Canal Cancer with Pagetoid Spread with Different Antitumor Effects of Chemotherapy on Different Metastatic Sites].

A 78-year-old man visited our hospital with a prolapsed hemorrhoid. He was referred to the dermatology unit due to the thickness and redness of the perianal skin. He was diagnosed as having extra mammary Paget's disease by skin biopsy. After a biopsy of the anal polyp was performed to investigate the primary site, he was diagnosed with early anal canal cancer with Pagetoid spread and underwent a radical operation. Abdominoperineal resection with skin(D2 prx D3 lymphadenectomy) was performed with perineal reconstruction using a gracilis muscle graft. Postoperative surveillance without adjuvant therapy was performed because the pathological stage was stage I. Two years and 2 months after surgery, multiple liver metastases were found, and the patient was diagnosed with multiple liver, bone, and lymph node metastases(K-ras and UGT1A1 wild type)on PET. XELOX plus bevacizumab was used as first-line treatment and the liver metastases showed remarkable shrinkage; however, disease progression occurred in the bone. IRIS plus bevacizumab was started as second-line therapy but grade 3 hematotoxicity was observed during the first course. After 4 courses, it was difficult to maintain the therapy due to toxicity and cancer-related pain. The liver metastases had almost disappeared but the patient died 11 months after the initiation of chemotherapy.

[Clinical Significance of Serum p53 Antibody Measurement in Colorectal Cancer Patients].

We investigated the significance of serum p53 antibody in colorectal cancer. The subjects included 239 patients with colorectal cancerwithout any history of priortr eatment. Patients with multiple cancerordouble cancerwer e excluded. Serum p53 antibodies and relationships with clinicopathological factors and prognosis were examined. The preoperative positive rate of serum p53 antibody was 28.9%. There was no relationship between p53 antibody and clinicopathological factors or degree of progression. Examination of a possible relationship with prognosis revealed that p53 antibody was not related to whether recurrence occurred, and there was no difference observed for survival rate. Examination of the cases in which serum p53 antibody was measured continuously after curative resection revealed that there was a tendency to find a low incidence of cancer recurrence in cases in which serum p53 antibody became negative postoperatively. Furthermore, cancer recurred at a low incidence in the cases in which p53 antibody was on the decrease postoperatively(p=0.0008). Thus, ourr esults show that serum p53 antibody in colorectal cancer cases is not related to clinicopathological factors or prognosis. However, in cases positive for preoperative serum p53 antibody, changes in postoperative levels of the titer could be a marker for recurrence.

[The Significance and Limitations of Curative Resection for Peritoneal Metastases of Colorectal Cancer].

The objectives of the present study were to investigate the treatment outcomes of curative resection in patients with peritoneal metastases from colorectal cancer and to clarify the significance and limitations thereof. The study included 38 patients with colorectal cancer who underwent curative resection of peritoneal metastases between 1996 and 2014. Peritoneal metastases were classified as follows: metachronous(n=9)and synchronous(n=29); P1(n=13)and P2(n=25); and ovarian(n=5). Thirty patients received postoperative chemotherapy, includingoxaliplatin -based regimens(n=14)and other regimens, such as 5-FU/Leucovorin(n=16). The 3-year survival rate amongall patients was 59.9%. There were no differences in survival rates accordingto gender, serum CEA levels, location, differentiation, depth of invasion of tumor, number of organs with distant metastases, severity of peritoneal metastasis, and types of postoperative chemotherapy. However, poor outcomes were observed in patients with more advanced lymph node disease and in patients with metachronous metastases compared to those with synchronous metastases. Although curative resection can contribute to long-term survival in colorectal cancer patients with peritoneal metastases, the therapeutic effect may be limited in patients with lymph node metastases and those with metachronous metastases.

[Preoperative Prognostic Nutritional Index in Elderly Patients Over 80 Years Old Who Underwent Curative Resection for Colorectal Cancer].

PATIENTS AND METHODS: From 2001, cases of 76 patients who underwent Cur A resection for colorectal cancer and preoperative prognostic nutritional index (PNI) assessment were analyzed in terms of clinicopathological factors, surgical outcomes, and prognosis. RESULTS: Median age of the 36 men and 40 women included in the study was 83 years. Median PNI was 42.1. Twenty-four patients had a PNI below 40. There were 36 patients with pulmonary dysfunction, and 60 patients with D2 or wider LN dissection. Median operating time was 177 minutes. Postoperative complications were reported in 48 patients and SSI in 15 patients. Postoperative hospital stay was 16 days. During a median 30 month follow-up period, there were 16 deaths, including 8 from cancer-related causes and 8 from other disease-related causes. Patients with a PNI below 40 often had fewerD2 LN dissections(p=0.082)and significantly shorter operating times(p=0.015). First gait and first flatus were observed significantly later(p=0.0051, 0.0307). There were no significant differences in postoperative complication rates, SSI, postoperative hospital stay, cause of death, or survival. CONCLUSIONS: In the elderly aged over 80-years-old, because those with a PNI below 40 underwent resection with fewer LN dissections and shorter operating time, there were no significant differences in postoperative complications, cause of death, or prognosis. These results suggest that PNI could be a selection factor for surgical procedure in elderly patients aged over 80 years.

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions in Large White pigs selected for higher peripheral blood immune capacity.

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions were compared between the immunity-selected Large White line and the non-selected Large White line. The selected Large White line showed a higher level of pulmonary MPS lesions compared with the non-selected Large White line. Subsequent to vaccination, the percentage of natural killer cells and T cells (CD3(+) CD4(+) CD8(-) and CD3(+) CD4(-) CD8(+) T cells) were significantly increased in the non-selected line but remained unchanged in the immunity-selected Large White line. Secretion of Mycoplasma hyopneumoniae vaccine-specific immunoblogulin G and phagocyte activity in peripheral blood were significantly higher in the immunity-selected Large White line than in the non-selected line. Expression of interleukin (IL)-4 and IL-6 messenger RNA in hilar lymph nodes was significantly lower in the immunity-selected Large White line than in the non-selected line. However, expression of IL-10 in all immune tissues was significantly higher in the immunity-selected Large White line. These results suggest that the selection for high immunity was not effective in increasing resistance to MPS lung lesions.

Immunogenic properties of Landrace pigs selected for resistance to mycoplasma pneumonia of swine.

Mycoplasma pneumonia of swine (MPS) lung lesions and immunogenic properties were compared between a Landrace line that was genetically selected for reduced incidence of pulmonary MPS lesions, and a non-selected Landrace line. The MPS-selected Landrace line showed significantly lower degrees of pulmonary MPS lesions compared with the non-selected Landrace line. When changes in immunity before and after vaccination were compared, the percentage of B cells in the peripheral blood of the MPS-selected Landrace line was significantly lower than that of the non-selected line. Furthermore, the concentration of growth hormone and the mitogen activity of peripheral blood mononuclear cells in the MPS-selected Landrace line showed significantly (P < 0.05) lower increases after vaccination than the non-selected line. Conversely, the concentration of peripheral blood interferon (IFN)-γ and salivary immunoglobulin A (IgA) after Mycoplasma hyopneumoniae vaccination was significantly higher in the MPS-selected Landrace line than in the non-selected line. Gene expression of toll-like receptor (TLR)2 and TLR4 was significantly higher in the MPS-selected Landrace line in immune tissues, with the exception of the hilar lymph nodes. The present results suggest that peripheral blood IFN-γ, salivary IgA TLR2, and TLR4 are important immunological factors influencing the development of MPS lesions.

Mycoplasma pneumonia of swine (MPS) resistance and immune characteristics of pig lines generated by crossing an MPS pulmonary lesion selected Landrace line and a highly immune capacity selected Large White line.

To understand the influence of crossbreeding on Mycoplasma pneumonia of swine (MPS) resistance and immune characteristics, two crossbred lines were characterized. One crossbred line, LaWa, was generated by crossing the MPS pulmonary lesion selected Landrace line (La) and the highly immune-selected Large White line (Wa). The second crossbred line, LaWb, was generated by crossing the La line and the nonselected Large White line (Wb). The crossbred LbWb line (nonselected Landrace line × nonselected Large White line) and the La line were used as controls. The LaWa and LaWb lines had an intermediate level of MPS lung lesions between La and LbWb lines, although the difference was not statistically significant. After stimulation with sheep red blood cells (SRBCs), the LaWb and LaWa lines showed immune characteristics similar to that of the La line; the number of monocytes in peripheral blood increased, while B cells, T cells, secretion of SRBC-specific immunoglobulin G, and interleukin (IL)-13 decreased. Additionally, the number of natural killer (NK) cells and the expression of IL-4 and IL-17 were significantly higher in the LaWb and LaWa lines, respectively. These data suggested that crossbreeding of La and Wa lines resulted in the inheritance of some of the selected immune responses.

[Immuno-Nutritional Factors Affecting the Incidence of Surgical Site Infection（SSI）after Rectal Cancer Surgery].

BACKGROUND: We analyzed immune nutritional factors that affected the incidence of SSI in rectal cancer surgery. METHOD: A total of 103 patients who underwent rectal cancer resection were enrolled in this retrospective study. The risk factors (DM, BMI<18.5, ≥25.0, PNI≤40, G/L>2, CONUT≥2, mGPS D) for SSI (Grade≥Ⅱ) were analyzed. RESULT: The factors that significantly affected SSI (in 13 cases) was PNI≤40 on univariate analysis. In the analysis adjusted by age and sex, mGPS D and PNI≤40 were significant factors. In the stepwise selection method, PNI≤40 was selected as an independent factor. CONCLUSION: As a risk factor for SSI after rectal cancer surgery, PNI≤40 and mGPS were risk factors.

[Short-Term Outcome of TAS-102 for Refractory Metastatic Colorectal Cancer].

We retrospectively analyzed 7 patients with refractory colorectal cancer treated with TAS-102 as salvage therapy. Subjects were 3 men and 4 women. The median age at initiation of TAS-102 was 71 years (range, 41-82 years). The number of target organs was 1 in 5 patients, 2 in 1 patient, and 3 in 1 patient. The median treatment courses were 2 courses (range, 1-6 courses). The reason for discontinuation was hematological toxicity in 1 patient, patients' wish in 3 patients, disease progression in 2 patients, and worsening of general condition in 1 patient. The median survival time since the first administration of TAS-102 was 9 months.

[Investigation of Intravenous Azithromycin Treatment Safety When Reducing Solvent for Intensive Care Unit Patients].

Intravenous azithromycin (AZM) was approved for use in December 2011 in Japan. In general, intravenous AZM injections are diluted to 1 mg/mL, with a total infusion volume of 500 mL to avoid phlebitis. Patients in intensive care units (ICUs) require small infusion volumes. We retrospectively evaluated the total AZM infusion volume in 65 ICU patients receiving AZM treatment from December 2011 to August 2014. Thirteen patients (20.0%) received a reduced volume [100 mL (5 mg/mL) or 250 mL (2 mg/mL)] using an infusion pump over 2 h. No peripheral phlebitis was observed in any patient. Based on this result, it is assumed that AZM can be safely administered to ICU patients even though the volume of solvent is reduced. AZM is widely recommended for the treatment of community-acquired respiratory infections and is used in patients with severe infections. Further investigation is required in additional patients to understand the effects of AZM volume reduction in greater detail.

[The Significance of Primary Tumor Resection in Unresectable Stage â £ Colorectal Cancer].

The significance of primary tumor resection for unresectable Stage Ⅳcolorectal cancer is controversial. In the present study, we examined cases of unresectable Stage Ⅳ colorectal cancer treated in our department. The subjects were 78 patients with unresectable Stage Ⅳ colorectal cancer who received either resection of the primary tumor, intensive chemotherapy, or both, between 2006 and 2012. The patients were divided into 2 groups: the group that received primary tumor resection (67 patients) and the non-resection group (11 patients). No differences were noted between a history of primary tumor resection and various clinicopathological factors, but the prognoses in the primary tumor resection group were favorable. The subjects were divided into 3 groups based on the selection of primary tumor resection and chemotherapy. The median survival time was 21.6 months, 11.8 months, and 8.1 months for patients who underwent chemotherapy after primary tumor resection (52 patients), patients who received primary tumor resection only (15 patients), and patients who received only chemotherapy (11 patients), respectively. The prognoses of patients who received primary tumor resection were favorable in comparison with those who received only chemotherapy. The results of the present study suggest the possibility that primary tumor resection can improve the prognoses of patients who have unresectable Stage Ⅳ colorectal cancer.

[A Case of Peritoneal Metastasis in Which Colostomy Was Useful for Restenosis after Stenting].

We report a case of restenosis after performing stenting twice for ileus caused by peritoneal dissemination that occurred after surgery for sigmoid colon cancer, in which colostomy was performed to improve the patient's QOL. The patient was a 58-year-old woman who underwent sigmoidectomy for sigmoid colon cancer. She presented with a peritoneal recurrence 3 times, and the third surgery was a non-curative resection. Chemotherapy was administered but was discontinued because of severe adverse events, and the patient was followed up with the best supportive care. An anastomotic stricture occurred 4 years after the initial surgery, and despite performing stenting twice, stenosis occurred 3 times within a few months. The third stenosis occurred shortly after the second episode, and colostomy was therefore performed. The patient died from cancer 4 months after colostomy without having another episode of stenosis. Although stenting is effective for patients with malignant colon stenosis, colostomy appears to be more effective for repeated post-stenting stenosis, when the patient is in an eligible general condition.