Anti-Inflammatory Effects of a Novel Nuclear Factor-κB Inhibitory Derivative Derived from Pyrazolo[3,4-d]Pyrimidine in Three Inflammation Models.

Nuclear factor-κB (NF-κB) plays a central role in inflammatory responses, and its physiologic functions are essential for cell survival and proliferation. Currently, drugs targeting NF-κB inhibition have not yet been applied in clinical practice. We investigated the physiologic effect of a novel NF-κB inhibitory compound, 1H-pyrazolo[3,4-d]pyrimidin-4-amine derivative (INH #1), on three inflammatory animal models. The pharmacokinetics were measured by liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. Acute hepatitis was induced by administrating lipopolysaccharide (LPS) and D-(+)-galactosamine hydrochloride followed by the analysis of survival time and inflammatory mediators. Collagen-induced arthritis (CIA) was induced by immunization with type II collagen (CII), and serum-transfer arthritis (STA) was caused by injecting K/BxN mice serum. Clinical and histologic scores were evaluated in both arthritis models. Immune cell subset analysis, CII-induced interferon-gamma (IFN-γ) production and proliferation, and measurement of anti-CII IgG antibodies were performed in the CIA model. In the acute hepatitis model, INH #1 suppressed tumor necrosis factor-α (TNF-α) production and prevented early death in a dose-dependent manner. INH #1 significantly attenuated arthritis scores and joint inflammation in both arthritis models. Additionally, in the CIA model, dendritic cells (DCs) in the regional lymph nodes were decreased in the treated mice and antigen-induced IFN-γ production and cell proliferation in splenocytes were inhibited, whereas the titers of anti-CII IgG antibodies were comparable regardless of the treatment. Here we revealed that INH #1 exerted anti-inflammatory effects in vivo via inhibition of inflammatory mediators and suppression of cellular immune responses. This compound could be a novel candidate for inhibition of NF-κB in certain inflammatory diseases. SIGNIFICANCE STATEMENT: A novel nuclear factor-κB (NF-κB) inhibitory compound, 1H-pyrazolo[3,4-d]pyrimidin-4-amine derivative (INH #1), which retains physiologically essential NF-κB bioactivity, suppressed inflammation in three different mouse models: the acute hepatitis model, the collagen-induced arthritis model, and the K/BxN serum-transfer arthritis model. These results suggest that this compound could be a novel and potent anti-inflammatory agent.

Chondroprotective effects of CDK4/6 inhibition via enhanced ubiquitin-dependent degradation of JUN in synovial fibroblasts.

OBJECTIVE: Targeting synovial fibroblasts (SF) using a cyclin-dependent kinase (CDK) 4/6 inhibitor (CDKI) could be a potent therapy for RA via inhibition of proliferation and MMP-3 production. This study was designed to elucidate the mechanism of chondroprotective effects on SFs by CDK 4/6 inhibition. METHODS: CDK4/6 activity was inhibited using CDKI treatment or enhanced by adenoviral gene transduction. Chondroprotective effects were evaluated using a collagen-induced arthritis model (CIA). Gene and protein expression were evaluated with quantitative PCR, ELISA and Western blotting. The binding of nuclear extracts to DNA was assessed with an electrophoresis mobility shift assay. RNA-Seq was performed to identify gene sets affected by CDKI treatment. RESULTS: CDKI attenuated cartilage destruction and MMP-3 production in CIA. In RASFs, CDKI impaired the binding of AP-1 components to DNA and inhibited the production of MMP-1 and MMP-3, which contain the AP-1 binding sequence in their promoter. CDK4/6 protected JUN from proteasome-dependent degradation by inhibiting ubiquitination. The RNA-Seq analysis identified CDKI-sensitive inflammatory genes, which were associated with the pathway of RA-associated genes, cytokine-cytokine receptor interaction and IL-17 signalling. Notably, the AP-1 motif was enriched in these genes. CONCLUSION: The mechanism of chondroprotective effects by CDK4/6 inhibition was achieved by the attenuation of AP-1 transcriptional activity via the impaired stability of JUN. Because the pharmacologic inhibition of CDK4/6 has been established as tolerable in cancer treatment, it could also be beneficial in patients with RA due to its chondroprotective and anti-inflammatory effects.

Effectiveness and safety of treat-to-target strategy in elderly-onset rheumatoid arthritis: a 3-year prospective observational study.

OBJECTIVES: To evaluate 3-year outcomes of following a treat-to-target (T2T) strategy targeting low disease activity for patients with elderly-onset RA (EORA) and to confirm safety profile of T2T. METHODS: Treatment was adjusted to target low disease activity with conventional synthetic DMARDs, followed by biologic DMARDs (bDMARDs) in 197 MTX-naïve EORA patients (mean age 74.9 years) with moderate-to-high disease activity. Non-implementation of T2T was evaluated at week 12, 24, 36, 52, 76, 104 and 128. To evaluate risks of using MTX, bDMARDs and glucocorticoids, 2122 periods of 3 months each were analysed using Bayesian hierarchical logistic regression models. RESULTS: Of the patients, 84.7% received methotrexate, 34.0% glucocorticoids with DMARDs and 41.6% bDMARDs during the observation period. Sixty-nine of the 197 patients failed to adhere to T2T because of comorbidities or the patient's own decision: 33 failed once, 19 twice, 10 three times and 6 four times or more. Simplified disease activity index (SDAI) remission and HAQ Disability Index (HAQ-DI) ≤0.5 at 3 years were achieved in 57.8% and 70.3% of the 128 patients adhering to T2T, and 34.8% and 43.5% of the 69 patients who did not adhere to T2T, respectively, and these were significantly different. Eighty-nine serious adverse events (SAEs) of any type were reported in 61 patients. MTX, bDMARDs and glucocorticoid were not associated with SAEs when adjusted for mean SDAI during the observation period and comorbidities at baseline. CONCLUSION: T2T strategy for EORA by using MTX and bDMARDs was effective with an acceptable safety profile. Adhering to T2T led to better outcomes.

[Laparoscopic Gastrectomy for Gastric Cancer with Adachi Type â ¥ Vascular Anomaly-A Case Report].

We report a case of laparoscopic gastrectomy for gastric cancer with an anomalous celiac trunk categorized as Type Ⅵ- Group 24 in the Adachi classification. Upper gastrointestinal endoscopy in an 81-year-old male revealed a shallow depressed lesion in the middle of the gastric body. Close examination led to diagnosis of cT1bN0M0, cStage Ⅰ gastric cancer, and laparoscopic distal gastrectomy was planned. Contrast-enhanced CT revealed no anomalous bifurcation of the hepatic artery, but the common hepatic artery ran on the dorsal side of the portal vein, branching from the superior mesenteric artery. Therefore, an Adachi Type Ⅵ-Group 24 celiac trunk anomaly was diagnosed. During surgery, the common hepatic artery could not be confirmed in guiding suprapancreatic lymph node dissection, and the portal vein was exposed. Anterior to the portal vein, nerves that are usually around the common hepatic artery continuously ran toward the hepatoduodenal ligament instead. Suprapancreatic lymph nodes were dissected, with the portal vein considered as the common hepatic artery. Adachi Type Ⅵ is a rare anomaly with an incidence of about 2%. Preoperative diagnosis enables safe and appropriate lymph node dissection.

The effects of bolus supplementation of branched-chain amino acids on skeletal muscle mass, strength, and function in patients with rheumatic disorders during glucocorticoid treatment.

OBJECTIVES: To test the effects of bolus supplementation of branched-chain amino acids (BCAA) on skeletal muscle mass, strength, and function in patients with rheumatic disorders taking glucocorticoid (GC). METHODS: Patients with rheumatic disorders treated with prednisolone (≥10 mg/day) were randomized to ingest additional daily 12 g of BCAA (n = 9) or not (n = 9) for 12 weeks. At baseline, and 4, 8, and 12 weeks, they underwent bioelectrical impedance analysis, muscle strength and functional tests, and computed tomography analysis for cross-sectional area of mid-thigh muscle. RESULTS: Disease activities of the patients were well controlled and daily GC dose was similarly reduced in both groups. Limb muscle mass was recovered in both groups. Whole-body muscle mass and muscle strength and functional mobility were increased only in BCAA (+) group. The effects of BCAA supplementation on recovering skeletal muscle mass were prominent in particular muscles including biceps femoris muscle. CONCLUSIONS: This trial is the first-in-man clinical trial to demonstrate that BCAA supplementation might be safe and, at least in part, improve skeletal muscle mass, strength, and function in patients with rheumatic disorders treated with GC.

[Two Cases of HER2-Positive Gastric Cancer with Multiple Liver Metastases Leading to Conversion Therapy with Chemotherapy].

Case 1: A 69-year-old man underwent chemotherapy with capecitabine plus cisplatin plus trastuzumab to treat advanced gastric cancer that was diagnosed as cStage IV adenocarcinoma(T3N2M1[P0, CYX, H1]). After 12 courses, liver metastases were absent on computed tomography images. The patient underwent total gastrectomy with D2 lymphadenectomy. It has been 22 months since the patient had gastrectomy without recurrence of the cancer. Case 2: A 70-year-old man underwent chemotherapy with capecitabine plus cisplatin plus trastuzumab for treatment of advanced gastric cancer that was diagnosed as cStage IV adenocarcinoma(T4aN1M1[P0, CY0, H1]). After 12 courses, regrowth of multiple liver metastases led to a treatment with weekly paclitaxel plus trastuzumab as a second-line chemotherapy. After 9 courses of second-line chemotherapy, multiple liver metastases were absent in computed tomography images. The patient underwent total gastrectomy with D2 lymphadenectomy. A small white nodule on the surface of S2 and S3 of the liver led to the patient receiving a partial liver resection. The pathological finding of the resected liver specimen was a metastasis of an adenocarcinoma. During continuous chemotherapy with weekly paclitaxel plus trastuzumab after gastrectomy, multiple liver metastases were revealed. The patient died 19 months after gastrectomy.

[Minilaparotomy in Pancreas-Preserving Total Duodenectomy for Familial Adenomatous Polyposis Associated with Spigelman Stage â £ Duodenal Polyposis].

INTRODUCTION: Reports on pancreas-preserving total duodenectomy (PPTD) for duodenal polyposis (DP) in familial adenomatous polyposis (FAP) patients are rare. We herein report a case of PPTD performed by minilaparotomy for DP in an FAP patient. CASE: A 27-year-old female FAP patient was diagnosed with Spigelman classification (SC) stage Ⅳ DP on gastroduodenoscopy. She underwent PPTD through a 7 cm upper abdominal incision. All polyps were confirmed as adenomas histopathologically. After 6 months of follow-up, complications related to total duodenectomy, such as weight loss, diabetes mellitus, and fatty liver have not been observed. DISCUSSION: SC stage Ⅳ refers to non-advanced cancers, and PPTD is basically prophylactic surgery. Therefore, pancreatoduodenectomy seems too aggressive for SC stage Ⅳ patients. PPTD by minilaparotomy is suitable for young female patients for its curability, esthetic outcome, and safety.

[A Case of NET G1 of Vater's Papilla in Familial Adenomatous Polyposis-Associated Duodenal Adenomatous Polyposis].

INTRODUCTION: Familial adenomatous polyposis (FAP) is characterized by the development of duodenal polyposis (DP), which later develops into colonic adenomatous polyps and, eventually, colorectal cancer. Neuroendocrine tumors (NET) are rare in FAP and reports of pancreas-preserving total duodenectomy (PPTD) to treat NET are limited. CASE: A 62-year-old. woman was previously diagnosed with FAP and she underwent a total colectomy and ileorectal anastomosis. Surveillance by upper gastrointestinal endoscopy revealed duodenal polyposis and a 35 mm flat, elevated tumor near the ampulla of Vater. She was diagnosed as having Spigelman stage Ⅳ DP and she underwent PPTD. Histopathology revealed a 7 mm NET G1 in the ampulla of Vater and multiple adenomas. DISCUSSION: Little is known about duodenal NET G1in FAP patients who undergo PPTD. Close follow-up is necessary.

[A Case of Pelvic Schwannoma, Mimicking Metastasis of Rectal Carcinoma].

INTRODUCTION: Schwannoma in the lateral lymph node region is extremely rare; however, this tumor has been reported to have relatively high SUV on PET-CT, suggestive of malignancy. CASE: A 67-year-old man with advanced lower rectal cancer had a small nodule with FDG accumulation (SUVmax 2.6) near the left internal iliac artery. His preoperative diagnosis was rectal cancer with lateral lymph node metastasis. He underwent super-low anterior resection with lateral lymph node dissection. Histopathological examination was conclusive for pT3 (A), with an Rt263D lymph node metastasis. Interestingly, a schwannoma was detected among the harvested lymph nodes. CONCLUSION: Although rectal cancer is known to involve pelvic lymph nodes, the role of preoperative diagnosis with FDG-PET is unclear. We should consider that schwannoma is associated with slight elevation of SUVmax and it may mimic lymph node metastasis.

[A Case of Carcinoma of the Uterine Body, Right Ovary, and Duodenum in a Patient with Familial Adenomatous Polyposis].

We report a case of 4 carcinomas of the uterine body, right ovary, and duodenum in a patient with familial adenomatous polyposis (FAP). Her mother's family line carries FAP. She underwent proctocolectomy with ileoanal anastomosis for FAP when she was 20 years old. She was diagnosed with carcinoma of the uterine body and right ovary, and underwent abdominal total hysterectomy, bilateral salpingo-oophorectomy, and omentectomy at 48 years of age. The pathological examination revealed endometrioid adenocarcinoma of the uterine body (Stage ⅠB) and endometrioid adenocarcinoma of the right ovary (Stage ⅠA). Her diagnosis was Stage Ⅳ according to the Spigelman classification of duodenal polyposis, and she underwent pancreas-preserving total duodenectomy at 50 years of age. The pathological examination was conclusive for 2 carcinomas in the adenoma, which were 20 mm and 25 mm in diameter, respectively. She has been well without any evidence of cancer recurrence 20 months after the pancreas-preserving total duodenectomy.

Angiogenesis in superficial esophageal squamous cell carcinoma: assessment of microvessel density based on immunostaining for CD34 and CD105.

OBJECTIVE: The esophagus is the only organ where changes in the superficial microvasculature from normal squamous epithelium to invasive cancer are evident by magnifying endoscopy. We investigated in detail the features of angiogenesis in early-stage esophageal cancer using CD34 and CD105 immunostaining, and also the correlation between angiogenesis and mononuclear cell infiltration. MATERIALS AND METHODS: Using 10 samples of normal squamous epithelium, 7 samples of low-grade intraepithelial neoplasia, and 45 samples of superficial esophageal cancer, we determined the microvessel density at hot spots showing positive staining for CD34 and CD105. We observed the histological features of CD34- and CD105-positive microvessels that corresponded to observations made by magnifying endoscopy. We then investigated the correlation between microvessel density and each histological situation or the grade of mononuclear cell infiltration. RESULTS: The histological features of CD34- and CD105-positive microvessels were able to explain the morphological changes in the microvasculature during cancer progression observed by magnifying endoscopy. The microvessel density for CD34 or CD105 was significantly correlated with each of the histological types (P < 0.001, rS = 0.51 and 0.76, respectively). Mononuclear cell infiltration at CD105 hot spots was most frequent in M1 and M2 cancer (94.7%). The correlation between the degree of mononuclear cell infiltration and microvessel density for CD105 staining was also significant (P < 0.001, rS = 0.49). CONCLUSIONS: The microvessel density based on CD34 and CD105 immunostaining can be used to corroborate observations of superficial esophageal squamous cell carcinoma made by magnifying endoscopy. Mononuclear cell infiltration may play an important role in angiogenesis at the early stage of cancer progression.

[Evaluation of immunity in elderly patients with unresectable gastric cancer receiving S-1/Lentinan combination chemotherapy].

PURPOSE: This retrospective study evaluated immunity in elderly patients with unresectable gastric cancer receiving S-1/ Lentinan combination chemotherapy. PATIENTS AND METHODS: This study included 10 patients aged≥70 years with unresectable gastric cancer who received S-1/Lentinan combination chemotherapy between October 2008 and December 2012. All patients gave written informed consent. Immune parameters for regulatory T cell(Treg)ratio, prostaglandin E2(PGE2), C3, CH50, and granulocyte/lymphocyte ratio were measured before chemotherapy initiation and at 7 weeks after it. Clinicopathological or immune parameters affecting overall survival(OS)were consequently evaluated. RESULTS: A high Treg ratio(p=0.02) and low PGE2(p=0.05)levels at 7 weeks after chemotherapy and a decrease in the Treg ratio(p=0.02)were found to be significant favorable factors affecting OS. CONCLUSION: The outcome of elderly patients with unresectable gastric cancer receiving S-1/Lentinan combination chemotherapy seemed to be correlated with the change in immunity.

[The efficacy and safety of FOLFIRI or combined FOLFIRI and bevacizumab treatment as second-line chemotherapy for metastatic colorectal cancer patients aged 75 years and older].

The safety and efficacy of FOLFIRI as second-line chemotherapy for metastatic colorectal cancer patients ≥ 75 years was retrospectively evaluated. We analyzed 106 patients, who received FOLFIRI or a combination of FOLFIRI and bevacizumab following oxaliplatin-based first-line chemotherapy. The clinical characteristics and outcome in elderly patients ≥75 years(elderly[EP]group; n=18)were compared with those in patients aged<75 years(control group; n=88). The number of patients treated by a combination of FOLFIRI and bevacizumab in the EP group was lower than that in the control group (27.8% vs 55.7%; p=0.03). The comparison revealed no significant differences in response rate, progression-free survival, overall survival, and the frequency of overall adverse events after the start of second-line chemotherapy, although the frequency of anemia(Bgrade 3, p=0.07)and alopecia(grade 1/2, p=0.054)tended to be higher in the EP group than in the control group. Although this study retrospectively analyzed a limited number of patients, our results indicate that the safety and efficacy of FOLFIRI as second-line chemotherapy for metastatic colorectal cancer are almost equal in patients ≥ 75 years and those aged<75 years.

[Detection of hepatic lymph node involvement by PET/CT in colorectal cancer patients with liver metastasis].

We investigated the clinical significance of hepatic lymph node (HN) metastasis in 50 patients with colorectal cancer metastases to the liver. Patients underwent preoperative positron emission tomography/computed tomography (PET/CT), and subsequently, resection of colorectal liver metastases with HN dissection from 2007 to 2013. In the present study, we evaluated whether PET/CT screening would be beneficial in diagnosing the HN metastases. Our results indicate that 2 histo- logically confirmed HN metastases were detected during preoperative PET/CT screening for 50 patients, of which only 1 patient received a positive diagnosis prior to surgery. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of predicting metastasis were 50%, 100%, 100%, 98%, and 98%, respectively. We conclude that PET/ CT screening is not feasible for detecting HN metastasis.

[Is PCI effective in predicting prognosis in the era of oxaliplatin-based chemotherapy in stage IV colorectal cancer with peritoneal dissemination ?].

PURPOSE: Peritoneal dissemination is expected to be present in P1, P2, and P3 according to the 8th edition of colon cancer handling convention. The peritoneal carcinomatosis index (PCI) plays a central role in classification of peritoneal metastases. PATIENTS AND METHODS: Peritoneal dissemination was examined during surgery in patients with colon cancer and peritoneal metastasis undergoing treatment with oxaliplatin-based chemotherapy. RESULTS: Correlation between PCI and P1, P2, and P3 was found. Moreover, using a PCI cut-off value of 6 for peritoneal dissemination, there was a significant difference in survival. CONCLUSION: Positive correlation was found between PCI and P classification of colon cancer. Regardless of the presence metastases to other organs, there is a possibility that PCI is effective in predicting prognosis of patients with peritoneal dissemination- positive colon cancer.

[Significance of the granulocyte-to-lymphocyte ratio as a prognostic predictor in patients with stage IV colorectal cancer undergoing chemotherapy].

The aim of this study was to evaluate the clinical significance of the granulocyte-to-lymphocyte(G/L)ratio as a prognostic predictor in patients with Stage IV colorectal cancer. A total of 83 patients who underwent oxaliplatin-based chemotherapy for Stage IV colorectal cancer were enrolled in the study. Univariate analysis indicated that the G/L ratio; number of involved organs(more than one organ); performance status ≥1; noncurability; and levels of hemoglobin, C-reactive protein, albumin, alkaline phosphatase, carbohydrate antigen 19-9, and lactate dehydrogenase before chemotherapy were significant prognostic factors. Noncurability was identified to be an independent, unfavorable factor for survival on multivariate analysis. When patients were divided into 2 groups according to the G/L ratio(the median was considered the cut-off value), the median survival time of patients with a high G/L ratio(≥3.0)was significantly worse than that of patients with a low G/L ratio(<3.0; 16.1 months vs 25.4 months, p=0.03). Further studies with more patients are required to examine whether the G/L ratio is a convenient biomarker affecting survival in patients with Stage IV colorectal cancer.

[Significant prognostic factors in stage IV gastric cancer].

The aim of this study was to evaluate clinicopathological variables, including the granulocyte-to-lymphocyte ratio(G/L ratio), as prognostic factors for Stage IV gastric cancer patients. A total of 70 patients treated for Stage IV gastric cancer were enrolled in this study. Univariate analysis indicated that age ≥70 years, performance status >2, resection not being performed, chemotherapy not being administered, high C-reactive protein(CRP)levels, and carbohydrate antigen 19-9 levels were significantly associated with poor survival. Multivariate analysis of these factors identified resection not being performed, chemotherapy not being administered, and high CRP levels as independent unfavorable factors of survival. Although the G/L ratio was not a prognostic factor for Stage IV gastric cancer patients in this study, further studies with greater number of patients are required to determine whether the G/L ratio is a significant biomarker associated with survival.

[Significance of preoperative radiotherapy and lateral lymph node dissection for locally advanced rectal cancer - a comparative study involving different hospitals].

The aim of this study was to compare the results between lateral lymph node dissection (LLND) alone, and preoperative radiotherapy (preRT) followed by LLND, for the treatment of locally advanced rectal cancer. This study assessed 44 consecutive patients with lower rectal cancer (clinical Stages II and III) without lateral lymph node metastasis by preoperative imaging at two hospitals. Twenty-five patients at one hospital received preoperative short-course radiation therapy (total 25 Gy) followed by a curative LLND operation (preRT group), and 19 patients at another hospital underwent a curative operation by LLND alone (non-preRT group). The 5-year locoregional relapse-free survival, disease-free survival, and overall survival rates were not different between the preRT and non-preRT groups. Although three patients in each group had lateral lymph node metastases, none developed pelvic sidewall recurrence. This study suggests that preRT followed by curative surgery with LLND for lower rectal cancers without lateral lymph node metastasis before surgery would not improve survival and local control rates.

[Efficacy of oxaliplatin-based adjuvant chemotherapy in stage II and III colorectal cancer with perforation].

This retrospective study evaluated the efficiency of oxaliplatin-based (Oxa-based) adjuvant chemotherapy for colorectal cancer with perforation. The study included 38 patients who underwent surgery for perforation associated with StageII and III colorectal cancer between April 1998 and March 2013. Patients with operative mortality were excluded from the study. The numbers of patients with StageII and III disease were 26 and 12, respectively. Adjuvant chemotherapy was received by 16 patients (62%) in the Stage II group. Seven patients (54%) in the Stage III group received adjuvant chemotherapy, of which 6 received Oxa-based adjuvant chemotherapy. The median relative dose intensity(range)of oxaliplatin was 83.3 (28.8- 100)%. Grade 3 adverse events were reported in 2 cases. The 3-year overall survival rates in the StageII and III groups were 60% and 79%, respectively. There was no significant difference in overall survival between the groups (p=0.28). Patients who received adjuvant chemotherapy had a 2.3-fold greater 3-year overall survival rate than those who did not receive adjuvant chemotherapy (80% vs 35%, p=0.01). The 3-year overall survival rates were 100% in patients who received Oxabased adjuvant chemotherapy (n=6), and 76%in patients who received adjuvant chemotherapy without oxaliplatin (n=17), with no significant difference between the groups (p=0.24). The Oxa-based adjuvant chemotherapy was tolerated equally well in Stage II and III colorectal cancer patients with or without perforation. Further studies are needed to compare the effect of oxaliplatin with other adjuvant chemotherapies.