Effectiveness of introducing a 20-gauge core biopsy needle with a core trap in EUS-FNA/B for diagnosing pancreatic cancer.

BACKGROUND: Endoscopic ultrasound-guided fine-needle aspiration/biopsy (EUS-FNA/B) is a standard method for pathological diagnosis of pancreatic solid lesions. The EchoTip ProCore 20G® (PC20), a 20-gauge biopsy needle with a forward-bevel core trap, has been available in Japan since 2015. METHODS: We compared the efficacy of the PC20 with that of the EchoTip ProCore 22G® (PC22) and Acquire 22G® (AC22) in EUS-FNA/B for diagnosing pancreatic cancer. This retrospective study included 191 patients with pancreatic cancer who underwent EUS-FNA/B using the PC20, PC22, or AC22 at our facility from April 2013 to October 2019. We investigated the patients' clinical characteristics and the diagnostic accuracy and safety of each needle. RESULTS: A sufficient stroke length of puncture was secured in all patients. The maximum length under EUS was shorter with the AC22 (22.1 ± 2.2 mm) than PC20 (30.6 ± 0.7 mm, p < 0.01) and PC22 (30.3 ± 0.8 mm, p < 0.01). The histological accuracy was 96.4% with the PC20 but only 58.8% with the PC22 (adjusted p (p-adj) < 0.0001) and 75.0% with the AC22 (p-adj = 0.06). The diagnostic accuracy of the combination of histology and cytology was 96.4% with the PC20, while it was 72.1% with the PC22 (p-adj < 0.0001) and 91.7% with the AC22 (p-adj > 0.99). One patient (0.9%) in the PC20 group developed mild pancreatitis, but no adverse events occurred with the other needles. CONCLUSIONS: The PC20 showed better diagnostic capability than the PC22. The diagnostic efficacy was similar between the PC20 and AC22. The high histological accuracy of the PC20 could be advantageous for lesions in which histological assessment is critical.

Bone regeneration is enhanced by the combined use of acid-electrolyzed functional water with hydroxyapatite/collagen composite.

Hydroxyapatite/collagen (HAP/Col) composite has a nanostructure and composition similar to that of natural bone. Herein, we have evaluated the beneficial effects of acid-electrolyzed functional water (FW) in combination with HAP/Col composite as an irrigation material in a rat calvarium defect model. The rats were divided into four groups: control, PBS irrigation; FW, FW irrigation; HAP/Col, filled with HAP/Col; FW + HAP/Col, FW irrigation prior to HAP/Col filling. Bone volume (BV) and bone mineral density (BMD) of the newly formed bone were analyzed by microcomputed tomography. The results indicated that the combined use of FW and HAP/Col significantly augmented both BV (12.25 ± 1.93 mm3 , control: 3.22 ± 0.55 mm3 , 6 weeks) and BMD (120.09 ± 14.76 cm3 /mg vs. control: 54.67 ± 7.20 cm3 /mg, 6 weeks) in a time-dependent manner, which might be attributed to the soluble factor-inducing ability of FW. Based on this assumption, bFGF concentration in peripheral blood was measured. bFGF concentration was significantly increased in the FW + HAP/Col group (68.25 ± 9.2 pg/ml vs. control: 21.70 ± 8.18 pg/ml, 6 hr). Real-time PCR demonstrated significant augmentation of MCSF (2.82 ± 0.59-fold), RANKL (2.51 ± 0.33-fold) and BMP7 (1.66 ± 0.25-fold) (bone regeneration-related genes) and PDGF (1.31 ± 0.15-fold), VEGF (3.27 ± 0.42-ld) and IL-8 (6.77 ± 2.02-fold) (angiogenic genes) mRNAs in the FW + HAP/Col group. Taken together, these results suggest that the combined use of FW and HAP/Col induces bone regeneration, presumably by inducing the factors contributing to bone regeneration and angiogenesis.

Pharmacokinetic-pharmacodynamic comparison of ceftriaxone regimens in acute cholangitis.

The most important factors determining the prognosis of patients with acute cholangitis (AC) are prompt biliary drainage and appropriate choice of antibiotics. This study was performed to evaluate whether dividing the number of doses based on the PK-PD theory contributes to better clinical outcome in the management of acute cholangitis. We measured ceftriaxone levels in blood and bile in 21 cases diagnosed with moderate-to-severe AC. Eleven cases were administered 2 g of ceftriaxone once-daily (group A) and 10 cases were given 1 g of ceftriaxone twice-daily (group B). The theoretical effect of ceftriaxone was evaluated by pharmacokinetic-pharmacodynamic (PK-PD) parameters. Clinical efficacy was evaluated by body temperature, white blood cell count and serum levels of C-reactive protein. Minimum level of ceftriaxone in serum (in mg/L) in groups A and B at 24 h after the first dose was 9.1 and 9.2, whereas that in bile was 2.9 and 2.5, respectively. The minimum inhibitory concentration (MIC) of ceftriaxone for all isolated bacteria was below the minimum serum and biliary concentration of ceftriaxone 24 h after the first administration (except for Enterococcus species). The MIC for isolated bacterial strains was <16 mg/L, which is the PK-PD breakpoint for ceftriaxone at 2 g/day. Both regimens showed clinical efficacy and did not contradict the effect predicted based on PK-PD.

Nickel chloride administration prevents the growth of oral squamous cell carcinoma.

The effect of NiCl2 on oral squamous cell carcinoma-derived cell line HSC3 was examined. Incubation with 1 mM NiCl2 significantly reduced the expression of MMPs at mRNA and protein levels. The in vivo orthotopic implantation model was established by injecting highly metastatic subcell line HSC3-M3 to nude mouse tongue. After 1 week of injection, mice were fed with or without 1 mM NiCl2-containing water for two to three weeks. Immunohistochamical examination revealed that MMP9 expression was drastically reduced in NiCl2-fed mice. By CT images, cancer mass was observed as a translucent area in control mice. In NiCl2-fed mice, much highly translucent area was observed within the translucent area. Histologically, this area corresponded to the necrotic area in the tumor mass. Real-time PCR analysis revealed the reduced expression of angiogenic factors such as IL-8 and VEGF mRNA in NiCl2-fed mice. To further examine the effect of NiCl2 on metastasis, human ß-globin gene expression in regional lymphnodes was compared. The ß-globin gene was totaly absent in NiCl2-fed mice. Moreover, various cancer metastasis-related genes were inhibited in NiCl2-fed mice by PCR array analysis. The results indicated that NiCl2 might be a promising new anti-cancer therapeutics for the oral cancer treatment.

Functional expression of TLR5 in murine salivary gland epithelial cells.

Toll-like receptors (TLR) recognize microbe-associated molecular patterns and induce the innate immune response. Among them, TLR5 recognizes the Gram-negative bacterial component flagellin. The aim of this study was to examine the expression of TLR5 in mouse salivary gland (SG). The SG was excised from 8- to 10-week-old female C57BL/6 mice. Salivary gland epithelial cells (SGECs) were purified and subjected to reverse transcription polymerase chain reaction (RT-PCR). Western blotting was performed to detect TLR5 expression at the protein level in several organs. The localization of TLR5 in SG was examined using immunohistochemical staining. The responsiveness of SGECs to flagellin was further examined by evaluating the induction of CXCL1 by real-time PCR and immunoprecipitation followed by Western blotting. TLR5 expression in SG was confirmed at the gene and protein levels. Immunohistochemical staining detected TLR5 in both acinic and ductal cells of the sublingual gland, but not in serous acinic cells of the submandibular gland. Although TLR5 was detected throughout the cytoplasm in ductal cells, positive staining was observed on the basal side of the mucous acinic cells. The purified SGECs responded to flagellin and induced the production of CXCL1. These findings suggest that TLR5 is functionally expressed in the SG and responds to its cognate ligand flagellin. (J Oral Sci 58, 317-323, 2016).

Electrolytically generated acid functional water inhibits NF-κB activity by attenuating nuclear-cytoplasmic shuttling of p65 and p50 subunits.

BACKGROUND: Human ß-defensin 2 (hBD2) gene expression is dependent on nuclear factor kappa B (NF-κB) activity. We have previously demonstrated that electrolytically generated acid functional water (FW) induces the expression of hBD2 in the human oral squamous cell carcinoma (OSCC) cell line Ca9-22. However, the induction was not dependent on NF-κB activity; in fact, FW inhibited NF-κB activity. Therefore, we hypothesized that FW might reduce spontaneous interleukin 8 (IL-8) secretion by Ca9-22 cells, which is heavily dependent on NF-κB activity. This study aimed at demonstrating the inhibitory effect of FW on NF-κB activity. METHODS: Ca9-22 cells were incubated with FW, and spontaneous IL-8 secretion was observed by enzyme-linked immunosorbent assay. Luciferase assay was performed using the 5'-untranslated region of the IL-8 gene. The steps of NF-κB activation blocked by FW were evaluated by localization of the NF-κB subunits p65 and p50 by immunofluorescence staining. Western blotting was further performed to confirm the changes in NF-κB subunit localization. RESULTS: The Ca9-22 cells spontaneously secreted IL-8, which was rapidly and drastically inhibited by FW treatment. The luciferase assay demonstrated the inhibitory action of FW, which was diminished by deletion of the NF-κB binding site from this construct. FW treatment altered the distribution of both the p65 and p50 subunits. P65, which was localized in the nucleus during the resting state, moved to the cytoplasm after FW treatment, whereas, p50, localized in the cytoplasm during the resting state, moved to the nucleus subsequent to FW treatment. CONCLUSIONS: The results from this study indicate that FW might inhibit spontaneous IL-8 secretion by redistribution of the NF-κB subunits within the cells.

Successful pregnancies treated with pioglitazone in infertile patients with polycystic ovary syndrome.

OBJECTIVE: To investigate the efficacy of pioglitazone on fecundity in infertile patients with polycystic ovary syndrome (PCOS) who are resistant to conventional ovulation induction such as clomiphene, dexamethasone, or metformin. DESIGN: A retrospective pilot study. SETTING: Department of Obstetrics and Gynecology in a general hospital. PATIENT(S): Nine infertile women with PCOS. INTERVENTION(S): Pioglitazone, 15-30 mg/day, up to 32 weeks. MAIN OUTCOME MEASURE(S): Pregnancy rate. RESULT(S): Seven of nine women became pregnant at an average of 11.3 weeks of initiation of pioglitazone. Four of seven pregnant cases conceived after the first successful induction of ovulation with the initiation of pioglitazone, and two cases conceived after the second cycle. Three women have already delivered, and there is one ongoing pregnancy. The other three cases ended in miscarriage during the sixth or eighth week of pregnancy. CONCLUSION(S): Pioglitazone may be effective in infertile patients with resistant PCOS.

Adding HPV16 testing to abnormal cervical smear detection is useful for predicting CIN3: a prospective study.

BACKGROUND: The aim of this prospective study was to estimate whether adding human papillomavirus 16 (HPV16) testing to abnormal cervical smears is useful in the prediction of cervical intraepithelial neoplasia 3 (CIN3). METHODS: Between October 1994 and May 1996, a total of 207 patients at the Akita University Hospital had abnormal smears. Of these patients, 153 patients with CIN1,2 or atypical squamous cells of undetermined significance (ASCUS) were enrolled in this study and followed until June 2001. At the initial visit, a cervical swab was collected for cytology and for HPV16 testing using nested polymerase chain reaction (PCR). When the HPV16 test was positive, HPV16 testing was performed every 3 to 6 months. We compared the prevalence of progression to CIN3 between the HPV16-positive group (n = 16) and the HPV16-negative group (n = 137). We also investigated the risk of progression to CIN3 associated with persistent HPV16 infection. RESULTS: At the end of the study period, four patients (25%) in the HPV16-positive group developed CIN3, and all of these patients were found to have persistent HPV16 infection during this period. Only three patients (2.2%) in the HPV16-negative group developed CIN3. CONCLUSIONS: The prevalence of progression to CIN3 in the HPV16-positive group was significantly higher than that in the negative group (p = 0.0023). The odds ratio of progression to CIN3 was 14.9 [95% confidence interval (CI) 2.98-74.4]. In particular, the risk of progression to CIN3 increased with persistent HPV16 infection. Adding HPV16 testing when abnormal cervical smears are detected promises to be useful for predicting CIN3.

Stromal vascularization in the endometrium during adenomyosis.

In adenomyosis, the two major frequent symptoms are hypermenorrhea and dysmenorrhea. The incidence of the former is said to be high, ranging from 36 to 70%. It has been found that vascular distribution of the endometrium in adenomyosis is diversely different from fertile patients without adenomyosis. For example, hysteroscopy revealed that approximately half of the patients have abnormal vascularization. In these patients, vascular distribution was generally irregular, and vessels were thick, dilated, and/or reticular in some patients. Moreover, morphometric analysis of the endometrium revealed that in the fertile women, the mean surface area, total surface area, and total number of capillaries, all increased significantly in the secretory phase compared to the proliferative phase. In contrast, the above parameters increased in the adenomyosis group in both the proliferative phase and secretory phase compared to the fertile women. In particular, the total surface area of capillaries per mm(2) markedly rose, by 11.6 times, compared to that of the proliferative phase in the fertile women. These findings suggest that regulatory factors involved in the vascular proliferation are diversely exaggerated, and that the abnormal vascularization of the endometrium is closely related with hypermenorrhea.

Involvement of catalase in the endometrium of patients with endometriosis and adenomyosis.

OBJECTIVE: To determine the distribution of catalase in eutopic and ectopic endometria in patients with endometriosis or adenomyosis. DESIGN: Retrospective randomized study. SETTING: Department of obstetrics and gynecology in a university hospital. PATIENT(S): Thirty-three patients with endometriosis, 36 with adenomyosis, and 47 fertile controls (total, 116 women). MAIN OUTCOME MEASURE(S): Semiquantitative immunostaining of endometrial cells obtained by biopsy sampling, followed by calculation of an evaluation nomogram score. RESULT(S): The score of catalase in the glandular epithelium of controls group fluctuated during the menstrual cycle; it was lowest in the early proliferative phase and peaked in the late secretory phase. In patients with endometriosis, catalase scores did not fluctuate during the cycle, and scores were high compared with controls throughout the menstrual cycle. Catalase scores did not vary in patients with adenomyosis, and scores in this group were consistently higher than those in patients with endometriosis throughout the cycle. CONCLUSION(S): Abnormal expression of catalase in the eutopic and ectopic endometrium strongly suggests pathologic involvement of free radicals in endometriosis and adenomyosis.

Suppression of natural killer cell activity by splenocyte transplantation in a rat model of endometriosis.

BACKGROUND: One immune characteristic of endometriosis is a decrease in natural killer (NK) cell activity. This study was performed to determine whether an abnormal immune reaction in an endometriosis animal model could be transferred to an animal of the same species. METHODS: An endometriosis model was prepared using 8 week old female rats by grafting a small section of one uterine horn onto the mesentery, followed 4 weeks later by removal of the spleen and remaining uterine horn. Splenocytes, that had been depleted of macrophages were injected via the tail vein, and NK cell activity of splenocytes was determined 4 days later. The uterus was simultaneously investigated immunohistochemically for immune cells. There was a control group (untreated; group 1), a control-splenocyte injection group (group 2), an experimental endometriosis model group (group 3) and an endometriosis model splenocyte injection group (group 4). RESULTS: Splenocyte NK cell activity was decreased in group 3 to 42.0% of that of group 1 and in group 4 to 38.9%. Immunohistologically, the number of NK cells in groups 3 and 4 markedly decreased to 62.0 and 55.1% of group 1 respectively. CONCLUSION: It was demonstrated that abnormal immunity caused by allograft of immune cells could recur in an endometriosis rat model.