[Robot Assisted Para-Aortic Lymphadenectomy in Gastric Cancer Surgery].

Para-aortic lymphadenectomy in gastric cancer surgery is a highly difficult surgical technique. In our hospital, we introduced robotic surgery in anticipation of the minimal invasiveness and advanced operability. We use a tunneling approach that progresses from the Treitz ligament to the peri-aorta. The transverse mesocolon is expanded with a tissue grasping clip, and the retroperitoneum is incised from the side of the Treitz ligament to approach the abdominal aorta and inferior vena cava. The No.16b1 and No.16a2 latero lymph nodes can be dissected with a good visual field. When it is judged that the visual field development of the No.16a2 inter-lymph nodes is poor, Kocher's operation is added. Since 2016, 18 patients have undergone para-aortic lymphadenectomy, 3 of whom underwent robotic surgery in our hospital. R0 resection was performed in all the cases, and 22.5 lymph nodes were dissected as No.16 lymph nodes(20.0 in all the cases included laparotomy). Although only a small number of patients were examined, robot-assisted para-aortic lymphadenectomy was considered safe.

[A Case of Combined Use of Mohs Paste and Radiation Therapy for Advanced Breast Cancer with Bleeding].

A 75-year-old female, at her initial presentation, the tumor occupied her entire right breast, with a foul-smelling exudate. A biopsy revealed ER-positive, HER2-negative breast cancer, and CT revealed multiple lung metastases. Paclitaxel and fulvestrant were administered sequentially, the bleeding from the right breast mass stopped and the mass flattened. But, as the tumor progressed, the right breast mass re-enlarged and began to re-bleed. Therefore, hemostatic treatment with Mohs paste was performed in parallel with tamoxifen. Hemostatic effect was observed for a while, but she gradually became refractory to Mohs paste, necessitating frequent blood transfusions. It was decided to discontinue systemic drug therapy and consider palliative treatment, and to perform radiation therapy in parallel with Mohs paste treatment for the purpose of local control. After radiation therapy, the bleeding has completely stopped and blood transfusion has not to be required for 6 months. Although systemic drug therapy has been discontinued at the patient's request, she is still alive. While systemic drug therapy was discontinued, we were able to confirm the pure local control effect of combination of radiation therapy and Mohs paste.

[Clinicopathological Characteristics of Early-Onset Colorectal Cancer(EOCRC)Cases in Our Hospital].

BACKGROUND: In recent years, the number of colorectal cancer in Europe and the U. S. has been decreasing, but there are increasing reports on the trend of early-onset colorectal cancer(EOCRC), which is a rare population with no established knowledge on its characteristics. SUBJECTS AND METHODS: Of 3,501 colorectal cancer cases treated at our hospital between April 2011 and December 2021, those aged 39 years and younger were included. RESULTS: There were 32 EOCRC cases, 11 males/21 females. The histological type was tub in 31 cases and por in 1 case. Postoperative adjuvant chemotherapy was administered in 14 patients, and 12 completed the scheduled course. Twenty nine patients underwent R0 resection, of which 6 patients had recurrence and 5 patients died of primary disease. In summary, although EOCRC patients were in good general condition and had a high completion rate of adjuvant chemotherapy, the relapse rate was high, suggesting the need for aggressive adjuvant chemotherapy and careful postoperative surveillance.

Detection of anti-SFTSV nuclear protein antibody in the acute phase sera of patients using double-antigen ELISA and immunochromatography.

Severe fever with thrombocytopenia syndrome (SFTS) is a life-threatening febrile illness that is caused by the SFTS virus (SFTSV). The diagnosis of SFTS is usually performed by detecting viral RNA. However, it has been reported that viral RNA is no longer detectable at 6-12 days after the onset of disease. In the current study, we have constructed a plasmid to express the recombinant nuclear protein (NP) based on the Japanese strain of SFTSV (J1). We developed a double-antigen enzyme-linked immunosorbent assay (ELISA) and immunochromatography (IC) assay using recombinant NP to detect antibody against SFTSV-NP. When we tested time-sequential samples from four patients with SFTS, antibody to SFTSV-NP were detectable not only during the recovery phase (days 10-622) but also during the acute phase (days 4-7) of the disease using both of a double-antigen ELISA and IC assay. SFTSV-RNA was detected until 8-11 days after onset, thus suggesting the coexistence of the virus and antibody during the acute phase of SFTS. These data suggest that assays for detecting antibody against SFTS-NP described in the current study may be applicable not only for the epidemiological studies but also for the diagnosis of SFTS.

Neutrophil-derived lactoferrin induces the inflammatory responses of rheumatoid arthritis synovial fibroblasts via Toll-like receptor 4.

OBJECTIVES: Damage-associated molecular patterns (DAMPs) are proposed to drive aberrant stimulation of Toll-like receptors (TLRs) in rheumatoid arthritis (RA) inflamed joints. In the current study we investigated the role of the neutrophil-derived lactoferrin (LTF), as an endogenous ligand for TLR4 in the inflammatory response of RA synovial fibroblasts (RASFs). METHODS: RASFs were stimulated with LTF, and the expressions of inflammatory cytokines in RASFs were measured. To clarify the TLR4 signalling pathway associated with LTF stimulation, a small molecular inhibitor of TLR4 (TAK242) and NF-κB inhibitor were used. The role of nuclear factor of activated T cells 5 (NFAT5) was identified using small interfering RNA. To reveal the interaction between NF-κB and NFAT5, cerulenin, which disrupts their interaction, was used. RESULTS: Stimulation of RASFs with LTF significantly increased the expressions of inflammatory cytokines and chemokines, such as IL-6, CCL20 and IL-8, in RASFs. LTF enhanced the mRNA expressions of these cytokines in RASFs stimulated by TNF-α. TAK242 almost completely inhibited the expressions of inflammatory cytokines and chemokines in RASFs stimulated by LTF. The NF-κB inhibitor partially repressed the expressions of IL-6 and IL-8 mRNAs induced by LTF, but not CCL20 mRNA expression. On the other hand, NFAT5 silencing decreased the expressions of CCL20 and IL-8 mRNAs induced by LTF, but not IL-6 mRNA expression. Cerulenin repressed the expressions of IL-6, CCL20 and IL-8 in RASFs stimulated by LTF. CONCLUSIONS: Neutrophil-derived LTF may play a role as an endogenous ligand for TLR4 expressed on RASFs. NFAT5-NF-κB enhanceosome might regulate the expressions of LTF-TLR4-responsive genes in RASFs.

The time-sequential changes of risk factors for adult T-cell leukemia development in human T-cell leukemia virus-positive patients with rheumatoid arthritis: a retrospective cohort study.

Objective: This study aimed to investigate the time-sequential changes of risk factors for adult T-cell leukemia (ATL) development in human T-cell leukemia virus type 1 (HTLV-1)-positive rheumatoid arthritis (RA) patients. Methods: HTLV-1 infection was screened using particle agglutination assay and confirmed via western blotting in 365 RA patients. Twenty-three HTLV-1-positive RA patients were included in the study cohort. Blood samples were obtained from these patients at each observation time point. The values of HTLV-1 proviral load (PVL) and serum soluble IL-2 receptor (sIL2-R), which are risk factors for ATL development, were measured using real-time PCR and enzyme immunoassay, respectively. Results: The study cohort comprised 79 person-years. The median HTLV-1 PVL and sIL2-R values of the HTLV-1-positive RA patients were 0.44 copies per 100 white blood cells (WBCs) and 406 U/mL, respectively. Three HTLV-1-positive RA patients showed a high PVL value. No remarkable changes were observed in the PVL and sIL2-R values during the observation period. However, one elderly HTLV-1-positive RA patient who had a high PVL value developed ATL during treatment with methotrexate and infliximab. Conclusion: A thorough clinical assessment of the risk factors for ATL development may be necessary in daily clinical practice for RA patients in HTLV-1-endemic areas in Japan.

Diversity of cell phenotypes among MT-2 cell lines affects the growth of U937 cells and cytokine production.

We previously reported the diversity of structure and integration sites of human T-cell leukemia virus type 1 (HTLV-1) provirus among different MT-2 cell lines. This raised the question as to whether cell phenotypes also differed among MT-2 cell lines. The influence of two different MT-2 cell lines (MT-2J and MT-2B) on the growth of the promonocytic leukemia cell line, U937, was investigated. Protein levels and mRNA expression of cytokines were also investigated. In addition, Western blot analysis of HTLV-1 regulatory proteins, Tax and HBZ, was also performed. Culture supernatant from MT-2B, but not MT-2J, cells showed marked suppressive effects on U937 cell growth. MT-2B showed high tumor necrosis factor (TNF)-α, TNF-ß, and interferon (IFN)-γ both in protein levels of the culture supernatant and mRNA levels of the cells. Analysis using recombinant cytokines indicated that the suppressive effects of MT-2B were due, at least in part, to high levels of TNF-ß and its synergic effects with IFN-γ in the culture supernatant. Protein levels of HTLV-1 Tax and HBZ were higher in MT-2B than those in MT-2J cells. These molecules have been reported to affect the cytokine production of HTLV-1 infected cells; therefore, the difference in these molecules may have accounted for the differences in cytokine production between MT-2J and MT-2B cells. Furthermore, because MT-2 cells showed a large variation of integrated HTLV-1 proviruses as well as cell phenotypes, it is important to exercise caution in the assessment and interpretation of experimental data from MT-2 cells.

Detection of microbial genes in a single leukocyte by polymerase chain reaction following laser capture microdissection.

Although isolation and identification of bacteria in a clinical specimen constitute essential steps for the diagnosis of bacterial infection, positive results of the bacterial culture are not always attained, despite observing the bacteria by Gram staining. As bacteria phagocytosed by the leukocytes are considered as the causative agents of infectious diseases, this study aims to introduce a new approach for the collection of only bacteria phagocytosed by the neutrophils in an animal model using laser capture microdissection (LCM) followed by the DNA identification using polymerase chain reaction (PCR). We inoculated representative bacteria (Escherichia coli and Staphylococcus aureus) into the abdominal cavities of specific pathogen-free C57BL/6 J mice. After 6 h inoculation, we collected the fluid samples from the peritoneal cavities of mice and demonstrated peritonitis by the increase of neutrophils. Then, we smeared the neutrophils on the membrane slides and collected single-cell phagocytosing bacteria by LCM. The supernatant of the cell lysate was supplied for the PCR reaction to amplify the 16S rRNA gene, and we validated the DNA sequences specific for the inoculated bacteria. In addition, PCR using specific primers for E. coli and S. aureus identified each species of bacteria. Hence, this study suggests that the combination of LCM and PCR could be a novel approach to determine bacteria in infectious diseases. Nevertheless, further investigation is warranted to test various additional bacterial taxa to demonstrate the general applicability of this method to clinical samples.

[A Case of Long-Term Survival of a Patient with Gastric Cancer with Synchronous Liver Metastasis and Portal Vein Thrombus after Multidisciplinary Treatment].

A 77-year-old man was diagnosed with gastric cancer with synchronous single liver metastasis and portal vein thrombus. His HER2 immunohistochemistry tumor score was 3+; therefore, we administered trastuzumab plus capecitabine plus cisplatin. After 2 courses of chemotherapy, we observed disappearance of the portal vein thrombus and tumor reduction as a partial response, according to the RECIST guidelines. We performed distal gastrectomy and right lobectomy; the therapeutic grades of the primary and metastatic tumors were 1a and 2, respectively. We administered postoperative chemotherapy, and no recurrent lesions have appeared 2 years after surgery. Multidisciplinary treatment for gastric cancer with liver metastasis might be a feasible and useful strategy.

Infection of defective human T-lymphotropic virus type 1.

In a previous study, we reported that an identical defective provirus had integrated into multiple sites of the genome of a representative human T-lymphotropic virus type 1 (HTLV-1) cell line, MT-2. A possible explanation for this may be the repeated infection of this defective provirus to a cell. Therefore, we attempted to determine whether a defective provirus could transmit during the co-culture of HTLV-1 uninfected human T-cell line, Jurkat, with MT-2 cells treated with mitomycin C. As a result, we established not only a cell line with the integration of one complete provirus, but also a cell line with the integration of one defective provirus. The rearrangement of the T-cell receptor -γ gene of these cell lines showed them to be derived from Jurkat cells. Both HTLV-1 Tax/Rex and HBZ RNA were detected in the cell line, which harbors a complete provirus. On the other hand, HBZ RNA and transcriptional product specific for the defective provirus were detected in the cell line, which harbors a defective HTLV-1 provirus only. These results suggested that a defective HTLV-1 provirus with large depletion of internal sequence could transmit to other cells. Moreover, the defective provirus can be transcriptionally active. This suggested the possibility that the defective HTLV-1 provirus found in the lymphocytes of HTLV-1 carriers and patients with adult T-cell leukemia may transmit to other T-cells in vivo. The results also suggested that defective provirus in HTLV-1 carriers could be functional and may play a role in leukemogenesis.

The diversity of the structure and genomic integration sites of HTLV-1 provirus in MT-2 cell lines.

A human T-lymphotropic virus Type 1 (HTLV-1) positive cell line, MT-2, derived from human cord leukocytes co-culturing with adult T cell leukemia/lymphoma (ATL) cells is commonly used in HTLV-1 research; however, the details of provirus integrated in MT-2 genome have not yet been characterized. In this study, five types of HTLV-1 proviral sequences were detected in 11 different sites of the genome in a reference MT-2 cell line. The five types of HTLV-1 proviral sequences were one complete proviral genome, two types of proviruses with deletion of large internal viral sequences (5.3 and 3.9 kB), one provirus with a large deletion (6.2 kB) from 5'LTR to position 6257, and one provirus of LTR only. The provirus with identical deletion of large internal viral sequence (5.3 kB) was found to be integrated into six different sites (chromosomes). A complete provirus and three of four types of defective provirus were consistently detected in two other MT-2 cell lines cultured in different laboratories. Not only Tax/Rex RNA and HBZ RNA, but also the transcriptional product for a specific defective provirus, were detectable in all three MT-2 cell lines. Because it has been reported that defective provirus is frequently detected in ATL cells, these results may be important in understanding the mechanism of HTLV-1 proviral polymorphism, which may be related to leukemogenesis. In addition, the large variation in integrated HTLV-1 proviruses makes it important for researchers to exercise caution in their assessment and interpretation of results using MT-2 cell lines.

Setting the stomach transection line based on anatomical landmarks in laparoscopic distal gastrectomy.

Laparoscopic distal gastrectomy has become widespread as a treatment for early gastric cancer in eastern Asia, but a standard method for setting the stomach transection line has not been established. Here we report a novel method of setting this line based on anatomical landmarks. At the start of the operation, two anatomical landmarks along the greater curvature of the stomach were marked with ink: the proximal landmark at the avascular area between the last branch of the short gastric artery and the first branch of the left gastroepiploic artery, and the distal landmark at the point of communication between the right and left gastroepiploic arteries. Just before specimen retrieval, the stomach was transected from the center of these two landmarks toward the lesser curvature. Then, about two-third of the stomach was reproducibly resected, and gastroduodenostomy was successfully performed in 26 consecutive cases. This novel method could be used as a standard technique for setting the transection line in laparoscopic distal gastrectomy.

Defective human T-lymphotropic virus type 1 provirus in asymptomatic carriers.

Few studies have specifically examined defective provirus in asymptomatic human T-lymphotropic virus Type 1 (HTLV-1) carriers and its relation to proviral DNA loads (PVLs). To assess the significance of defective provirus in asymptomatic carriers, we examined PVLs in peripheral blood mononuclear cells of 208 asymptomatic HTLV-1 carriers. The mean PVLs determined using primers for the pol region were less than that for the pX region in these carriers. Analysis of seven carriers with high PVLs for the pX region but lower PVLs for the pol region showed that four had single nucleotide polymorphisms of proviral genomes for the pol region and three had HTLV-1-infected cells with defective provirus. Three carriers with defective provirus showed high PVLs at their initial screens, and PVLs increased after a 10- to 12-year interval in two carriers. Southern blot assay showed clonal expansion of HTLV-1-infected cells, and the predominant clones changed during the observation period. These data suggest that although HTLV-1-infected cells with defective provirus may have a growth advantage, the predominant clones of HTLV-1-infected cells do not always survive for many years in asymptomatic carriers.

Evaluation of tracheal intubation in several positions by the Pentax-AWS Airway Scope: a manikin study.

PURPOSE: In emergency situations, rescuers must occasionally secure the airway while the patient is in a restricted position rather than in the ideal supine position. We hypothesized that the Pentax-AWS Airway Scope (AWS) may be useful for emergent tracheal intubation in such positions. METHODS: Thirteen non-anesthesia residents performed tracheal intubation on a simulated manikin in the supine (Supine), left-lateral decubitus (Left-LT), right-lateral decubitus (Right-LT), prone (Prone), and sitting (Sitting) position, respectively, to assess AWS performance. RESULTS: Intubations were successful in all five positions. The time needed to secure the airway did not differ significantly between the Left-LT and Supine positions. Doctors required significantly more time to secure the airway in the Prone, Sitting, and Right-LT positions than in the Supine position. Visual analog scale (VAS) scoring of the subjective difficulty of laryngoscopy was lower in the Supine position rather than in the Right-LT, Prone, and Sitting positions. The VAS score of subjective difficulty of tracheal tube passage through the glottis was significantly higher in the Sitting position than in the other four positions. CONCLUSION: Although tracheal intubations with AWS in all five positions tested were successful, intubation with the patient in the Sitting, Right-LT, and Prone positions was more difficult and required more time than that in the Supine position.

Multiple integrations of human T-lymphotropic virus type 1 proviruses in the engrafted cells from the asymptomatic carriers in NOD/SCID/gammacnull mice.

OBJECTIVES: Successful engraftment of human T-lymphotropic virus type 1 (HTLV-1)-infected cells and a marked increase of proviral DNA loads (PVLs) in non-obese diabetic/severe combined immunodeficient (NOD/SCID)/gammac(null) (NOG) mice have been reported. Whether the increased PVL in transplanted mice is due to the new infection of HTLV-1 was examined. METHODS: Mononuclear cells from 3 NOG mice with primary engraftment from asymptomatic HTLV-1 carriers were transplanted into a second group of NOG mice. HTLV-1 PVL, proviral integration by fluorescence in situ hybridization assay, expression of viral antigen, and T-cell clonality were analyzed. RESULTS: The PVLs in the secondarily transplanted NOG mice were significantly higher than those of primarily transplanted NOG mice. Multiple signals of HTLV-1 proviruses in the nucleus of the infected cells were revealed by fluorescence in situ hybridization analysis. Expression of HTLV-1 tax/rex mRNA and antigen was observed. The variety of T-cell clones was limited in the transplanted NOG mice. CONCLUSIONS: Multiple proviral integrations were considered to be due to the new infection from HTLV-1-infected cells to the other cells. Only a certain fraction of T cells seemed to have selectively survived in NOG mice after engraftment.

Comparison of tracheal intubation by the Macintosh laryngoscope and Pentax-AWS (Airway Scope) during chest compression: a manikin study.

To compare tracheal intubation with the Pentax Airway Scope (AWS) and the Macintosh laryngoscope (McL) during chest compression, 25 anesthesiologists (including 12 specialists having >5 years of experience and 13 trainees having <2 years of experience) performed tracheal intubation using either the McL or the AWS, with or without chest compression, on a manikin. Using the McL, both specialists and trainees took a significantly longer time (P < 0.01) to secure the airway with chest compression (17.3 +/- 3.7 and 22.5 +/- 8.0, respectively) and than without chest compression (11.3 +/- 2.9 and 13.9 +/- 4.4 s, respectively). No significant difference was observed in time needed to secure the airway using the AWS with or without chest compression in both groups. From the standpoint of experience, time to complete intubation for specialists using the McL during chest compression was significantly shorter than that for trainees. In contrast, the difference in time to complete intubation with the AWS during chest compression was not significantly different between the two groups. Based on these results, we conclude that the use of the AWS may reduce the time needed to secure the airway during chest compression.

[Anesthetic management for pneumonectomy in a patient with cardiac amyloidosis].

Cardiac amyloidosis may cause restrictive cardiomyopathy associated with heart failure, conduction disorder and ischemic heart disease. Therefore, patients with amyloidosis require careful hemodynamic monitoring in perioperative period. A 63-year-old man with cardiac amyloidosis was scheduled for pneumonectomy. His transthoracic echocardiography assessment showed a hypertrophic interventricular septum and slight decreased ejection fraction of 55%, but left ventricular (LV) diastolic function was decreased. Pulse Doppler for mitral valve inflow showed that the early peak velocity/atrial peak velocity (E/A) ratio was 0.9, the deceleration time (DT) was 163 msec and the early diastolic mitral annular tissue velocity (E') was 4 cm x sec(-1). These data suggested a pseudonormalization state. We performed careful monitoring using arterial pressure-based cardiac output (APCO), central venous oxygen saturation (ScvO2) and transesophageal echocardiography. There were no severe complications such as circulatory collapse and arrhythmia in the perioperative period.

Coexistent anaplastic and differentiated thyroid carcinoma: an immunohistochemical study.

The aim of the present study was to clarify the underlying molecules that might contribute to the highly aggressive behavior of anaplastic thyroid carcinoma. We selected 5 cases of anaplastic thyroid carcinoma that had a differentiated area to determine differences in the molecules of undifferentiated and differentiated cancer cells. We immunohistochemically examined the localization of nuclear antigen (Ki-67), proliferating cell nuclear antigen (PCNA), p53, apoptotic protease-activating factor-1 (Apaf-1), CD26, galectin-3, E-cadherin, and CD147. We found increased Ki-67, PCNA, and p53 labeling indices; decreased levels of Apaf-1, CD26, galectin-3, and E-cadherin; and overexpression of CD147 in the undifferentiated area compared with the differentiated area. These findings indicate high proliferative properties, suppression of apoptosis, disruption of cell-cell interaction, and induction of matrix metalloproteinases in the undifferentiated areas. Thus the molecules examined might be useful for evaluating the aggressive nature of this tumor and the prognosis.

Coexistent Anaplastic and Differentiated Thyroid Carcinoma.

The aim of the present study was to clarify the underlying molecules that might contribute to the highly aggressive behavior of anaplastic thyroid carcinoma. We selected 5 cases of anaplastic thyroid carcinoma that had a differentiated area to determine differences in the molecules of undifferentiated and differentiated cancer cells. We immunohistochemically examined the localization of nuclear antigen (Ki-67), proliferating cell nuclear antigen (PCNA), p53, apoptotic protease-activating factor-1 (Apaf-1), CD26, galectin-3, E-cadherin, and CD147. We found an increased Ki-67, PCNA, and p53 labeling indices; decreased levels of Apaf-1, CD26, galectin-3, and E-cadherin; and overexpression of CD147 in the undifferentiated area compared with the differentiated area. These findings indicate high proliferative properties, suppression of apoptosis, disruption of cell-cell interaction, and induction of matrix metalloproteinases in the undifferentiated areas. Thus the molecules examined might be useful for evaluating the aggressive nature of this tumor and the prognosis.

Mutation spectrum induced by dihydropyrazines in Escherichia coli.

Dihydropyrazine (DHP), which induces mutagenesis in E. coli, was investigated. From analyzing mutations in the chromosomal rpoB gene, the mutation spectrum in uvrB strain revealed the different behavior on exposure to two DHP derivatives 3-hydro-2,2,5,6-tetramethylpyrazine (HTMP), and 2,3-dihydro-5,6-dimethylpyrazine (DHDMP). A higher level of DHP-induced mutation was observed, with base substitutions at G : C pairs predominant. HTMP and DHDMP increased the frequency of G : C to T : A transversions. HTMP increased the frequency of G : C to A : T transitions, than did DHDMP. These findings suggest that DHPs prefer to attack the G : C pair and that different DHP derivatives may prefer distinct mutagenic base pairs; and further, that nucleotide excision repair may be involved in the repair of DHP-induced mutations.