Tumor-infiltrating B cells and T cells correlate with postoperative prognosis in triple-negative carcinoma of the breast.

BACKGROUND: In this study, we investigated CD20+ TILs in triple-negative breast cancer (TNBC) and their relationship with T lymphocyte subsets (CD4+, CD8+, CD25+, and FOXP3+), including their combined prognostic value using an immunohistochemical staining method. METHODS: We investigated 107 patients with TNBC for whom a full-face section stained by hematoxylin and eosin between 2006 and 2018 at Dokkyo Medical University Hospital was available. RESULTS: The strongest association of infiltrating CD20+ TILs was with CD4+ TILs. There was a significant relationship between CD20+ and CD4+ TILs (r = 0.177; p < 0.001), CD8+ TILs (r = 0.085; p = 0.002), and FOXP3+ TILs (r = 0.0043; p = 0.032). No significant relationships were observed between the CD20+ and CD25+ TILs (r = 0.012; p = 0.264). Multivariate analysis revealed that only the CD20+/FOXP3 ratio was an independent factor for relapse-free survival (p < 0.001) and overall survival (p < 0.001). Patients with tumors highly infiltrated by CD4+, CD8+, and CD20+ TILs had a good prognosis. In contrast, those with tumors weakly infiltrated by CD20+ TILs but highly infiltrated by CD25+ and FOXP3+ TILs had a poor prognosis. CONCLUSIONS: CD20+ TILs may support an increase in CD4+ and CD8+ TILs, which altered the anti-tumor response, resulting in a positive prognosis. CD20+ TILs correlated with FOXP3+ Treg lymphocytes, which were reported to be correlated with a poor prognosis. Our study suggested that TIL-B cells have dual and conflicting roles in TIL-T immune reactions in TNBC.

[Preoperative C-reactive protein predicts recurrence after curative resection in advanced gastric cancer patients receiving postoperative adjuvant chemotherapy using S-1].

New chemotherapeutic regimens such as S-1 plus docetaxel, S-1 plus oxaliplatin and capecitabine plus oxaliplatin are reported to be effective and safe as postoperative adjuvant chemotherapy (PAC) for advanced gastric cancer (GC) patients. Although the use of these PACs is increasing, it is still unclear how to choose the best regimen for advanced GC patients. Therefore, we aimed to investigate which clinical characteristics are associated with recurrence after curative surgery in patients receiving S-1 as PAC. Thirty-nine patients who received a PAC regimen with S-1 for more than 1 year after curative surgery for advanced GC were enrolled. Univariate and multivariate analyses using the Cox proportional hazard model were performed to detect clinical characteristics that correlated with recurrence. Patients were divided into two groups, recurrence, and non-recurrence, and receiver operating characteristic (ROC) curve analysis was used to identify the cut-off values. Kaplan-Meier analysis and the log-rank test were used for comparison of relapse-free survival (RFS). Fifteen patients had a recurrence after surgery (38.5%, 15/39). Multivariate analysis using clinical characteristics revealed that preoperative C-reactive protein (CRP) (>0.3/≤0.3, mg/dL) (HR 10.73;95% C.I., 1.824-63.14;P=0.009) was significantly associated with recurrence. Kaplan-Meier analysis and the log-rank test demonstrated that preoperative CRP (>0.3/≤0.3, mg/dL) was also significantly associated with RFS (P<0.001). Therefore, preoperative CRP is significantly associated with recurrence and RFS after curative surgery in advanced GC patients receiving S-1 as PAC.

[A Case of Locally Advanced Breast Cancer Successfully Treated with Local Control That Achieved a Pathological Complete Response after Bevacizumab and Paclitaxel Combination Chemotherapy].

A 68-year-old woman with a 5-year history of an untreated left breast tumor presented to our hospital. She was admitted for untreated diabetes and severe anemia. The cause of the anemia was bleeding from the tumor, and she was referred to our department. She was diagnosed with T4bN0M0, stage IIIb breast cancer. First, we initiated hormonal therapy. However, the tumor did not decrease in size. We then administered chemotherapy. The tumor markedly decreased in size, and mastectomy and axillary lymph node dissection were performed. The response was a pathological complete response. She is currently undergoing hormonal therapy at the time of this writing.

Combination of platelet count and neutrophil to lymphocyte ratio is a useful predictor of postoperative survival in patients undergoing surgery for gastric cancer.

OBJECTIVE: To investigate the clinical utility of the Combination of Platelet count and Neutrophil to Lymphocyte Ratio (COP-NLR) for predicting the postoperative survival of patients undergoing surgery for gastric cancer (GC). METHODS: The COP-NLR was calculated on the basis of data obtained on the day of admission as follows: patients with both an elevated platelet count (>300 × 10(3) /µl) and an elevated NLR (>3) were allocated a score of 2, and patients showing one or neither were allocated a score of 1 or 0, respectively. Five hundred forty-four patients for whom data were sufficient to allow analysis of the relationship between clinicolaboratory characteristics and postoperative survival were enrolled. RESULTS: Multivariate analysis using the 12 clinical characteristics selected by univariate analyses revealed that the COP-NLR was associated with OS (hazard ratio, 1.781; 95% C.I., 1.094-2.899; P = 0.020) along with age, tumor type, lymph node metastasis and albumin level. Kaplan-Meier analysis and log rank test demonstrated significant differences in both OS and cancer-specific survival among patients with COP-NLR 0, 1, and 2 (P < 0.001). CONCLUSION: The COP-NLR is able to predict postoperative survival of patients with GC and classify such patients into three independent groups before surgery.

A novel approach for sentinel lymph node identification using fluorescence imaging and image overlay navigation surgery in patients with breast cancer.

BACKGROUND: We reported a novel technique of sentinel lymph node (SLN) identification using fluorescence imaging of indocyanine green injection. Furthermore, to obtain safe and accurate identification of SLN during surgery, we introduce the image overlay navigation surgery and evaluate its efficacy. METHODS: This study enrolled 50 patients with a tumors <2 cm in diameter. Initially, we obtained three-dimensional (3-D) imaging from multidetector-row computed tomography (MD-CT) by volume rendering. It was projected on the patient's operative field with the clear visualization of lymph node (LN) through projector. Then, the dye of indocyanine green (ICG) was injected subdermally in the areola. Subcutaneous lymphatic channels draining from the areola to the axilla were visible by fluorescence imaging immediately. Lymphatic flow was reached after LN revealed on 3-D imaging. After incising the axillary skin on the point of LN mapping, SLN was then dissected under the guidance of fluorescence imaging with adequate adjustment of sensitivity and 3-D imaging. RESULTS: Lymphatic channels and SLN were successfully identified by Photodynamic eye (PDE) in all patients. And the sites of skin incision also were identical with the LN being demonstrated by 3-D imaging in all patients. The mean number of SLN was 3.7. The image overlay navigation surgery was visually easy to identify the location of SLN from the axillary skin. There were no intra- or postoperative complications associated with SLN identification. CONCLUSIONS: This combined navigations of fluorescence and 3-D imaging revealed more easy and effective to detect SLN intraoperatively than fluorescence imaging alone.

Prognostic value of tumor-infiltrating B lymphocytes and plasma cells in triple-negative breast cancer.

BACKGROUND: Recent investigations have demonstrated that the tumor microenvironment, including tumor-infiltrating lymphocytes (TILs), is an important factor in tumor growth and development. While the prognostic correlation of tumor-infiltrating T cells has been widely studied in breast cancer, that of tumor-infiltrating B cells and plasma cells has not received so much attention, especially in triple-negative breast cancer (TNBC). METHODS: We investigated 114 patients with TNBC who had surgery between 2006 and 2019 at Dokkyo Medical University Hospital. Intratumoral (i) TILs were considered to be lymphocytes within cancer cell nests and directly infiltrating tumor cells. Similarly, stromal (s) TILs were considered to be lymphocytes within the tumor stroma, but not directly infiltrating tumor cells. CD20 + , CD38 + and CD138 + staining was determined by estimating the number of positive B cells. RESULTS: sCD20 + TILs had prognostic significance for relapse-free survival (RFS) (p = 0.043) and overall survival (OS) (p = 0.027). The sCD38 + TILs were significantly related to favorable RFS (p = 0.042). iCD38, iCD138, and sCD138 was not significantly correlated with RFS (p = 0.065, p = 0.719, p = 0.074) or OS (p = 0.071, p = 0.689, p = 0.082). CONCLUSIONS: The present study demonstrated that a high density of sCD20 + TILs was significantly related to favorable prognosis in both RFS and OS. Increased sCD38 + TILs in TNBC were correlated with a significantly favorable prognosis in RFS. These results indicate that TILs-B may have a profound influence on the clinical outcome of TNBC.

CD68- and CD163-positive tumor-associated macrophages in triple negative cancer of the breast.

Tumor-associated macrophages (TAMs) have recently been reported as an important factor in tumor growth and the progression of cancer. The prognostic significance of localizations and densities of TAMs in triple negative cancer (TNC) of the breast is not well understood. The aim of this study was to assess the localizations and densities of the TAMs subtype in TNC and examine their clinicopathological features. The study was based on 107 TNC cases operated on at Dokkyo Medical University Hospital using the pan-macrophage marker CD68 and the M2 macrophage marker CD163 in the tumor stroma (TS) and tumor nest (TN), respectively, and examined the clinicopathological significance. Multivariate Cox regression analyses revealed that age and CD163+ TAMs in both the TS and TN were independent prognostic factors for relapse-free survival and overall survival. No correlation was found between the number of CD68+ cells or the CD163/CD68 ratio either in TS or TN, or clinicopathological features. Our study found that infiltration of CD163+ TAMs, rather than CD68+, in both TS and TN was associated with poor prognosis in TNC patients by multivariate analysis. This suggests that CD163+ TAMs may affect the prognosis of TNC by not only regulating the immune reaction by TAMs in TS, but also because of their direct influence on TN.

The clinical influence of the preoperative lymphocyte-to-monocyte ratio on the postoperative outcome of patients with early-stage gastrointestinal cancer.

AIM: The lymphocyte-to-monocyte ratio (LMR) is useful for predicting the prognosis of patients with gastric cancer (GC) and those with colorectal cancer (CRC) undergoing surgery. The relationship between the LMR and postoperative outcome of patients with early-stage gastrointestinal cancers such as stage I GC and CRC remains unclear. METHODS: We retrospectively evaluated 323 stage I GC and 152 stage I CRC patients undergoing surgery. Univariate and multivariate analyses using the Cox proportional hazards model were performed to identify the clinical characteristics associated with overall survival (OS), and the cut-off values of these variables were determined by receiver operating characteristic analysis. The Kaplan-Meier method and log-rank test were used for postoperative survival comparisons according to the LMR (GC: LMR < 4.2 vs ≥4.2; CRC: LMR < 3.0 vs ≥3.0). RESULTS: Univariate and multivariate analyses revealed that OS was significantly associated with the LMR (<4.2/≥4.2) (HR, 2.489; 95% CI, 1.317-4.702; P = 0.005), as well as age (>75/≤75 years) (HR, 3.511; 95% CI, 1.881-6.551; P < 0.001) and albumin level (≤3.5/>3.5 g/dL) (HR, 3.040; 95% CI, 1.575-5.869; P = 0.001), in stage I GC patients. Survival analysis demonstrated a significantly poorer OS in stage I GC patients with a LMR < 4.2 compared with ≥4.2 (P < 0.001). In stage I CRC patients, despite a significant difference in OS according to the LMR (<3.0 vs ≥3.0) (P = 0.040), univariate analysis revealed no significant association between the LMR and OS. CONCLUSION: LMR is a useful predictor of the postoperative outcome of stage I GC patients treated surgically.

Prognostic impact of a tumor-infiltrating lymphocyte subtype in triple negative cancer of the breast.

BACKGROUND: Tumor-infiltrating lymphocytes (TILs) have recently been reported as an important factor in the tumor microenvironment and influence the growth and progression of cancer. However, the relationship between immune cell subpopulations, such as CD4+, CD8+, and FOXP3+, in breast cancer, especially in triple negative carcinoma (TNC), remains unclear. METHODS: The subjects were 107 patients with TNC that were surgically resected at Dokkyo Medical University Hospital between 2006 and 2018. The expression of CD4+, CD8+, and FOXP3+ was evaluated in TILs and expressed as the numbers of positive cells. RESULTS: Univariate analysis revealed that the TILs were not prognostically significant. In multivariate analyses, increased infiltration of intratumoral (i) CD4+ TILs was found to have a good prognosis in relapse-free survival (RFS). In contrast, a high stromal CD8+ TILs level was found to be a favorable prognostic factor in RFS (p = 0.038) and overall survival (OS) (p = 0.046). A low sFOXP3 + TILs level was significantly associated with favorable RFS (p < 0.001) and OS (p = 0.029). CONCLUSIONS: The present study demonstrated no difference in TILs and survival in TNC. However, there was a significant correlation in prognosis with levels of iCD4+, sCD8+, and sFOXP3 + TILs in TNC. The difference in TNC clinical outcome may be due to the subtype of the infiltrating TILs.

High Enrichment of Nucleobase-modified Aptamers in Early Selection Rounds by Microbeads-assisted Capillary Electrophoresis SELEX.

Nucleobase-modified aptamers are attractive candidates for diagnostic and therapeutic agents due to the high affinity, stability and functionality. However, since even conventional SELEX requires many selection rounds, acquisition of modified aptamers is much more laborious. Herein, microbeads-assisted capillary electrophoresis (MACE)-SELEX was applied against thrombin using the indole-modified DNA library. After only three selection rounds, we successfully enriched the modified aptamers and they showed slower off-rate than reported aptamers, suggesting MACE-SELEX is a promising approach for rapid identification of modified aptamers.

Oestrogen receptor-negative/progesterone receptor-positive phenotype of invasive breast carcinoma in Japan: re-evaluated using immunohistochemical staining.

BACKGROUND: The existence of progesterone receptor (PgR) expression in oestrogen receptor (ER)-negative breast carcinoma is controversial. Here, we re-evaluated ER-negative/PgR-positive (ER-/PgR+) carcinoma cases by immunohistochemical staining (IHC). MATERIALS AND METHODS: We selected patients who underwent surgery for primary breast carcinoma from our databases at Dokkyo Medical University Hospital and Kameda General Hospital. Among the 9844 patients, the largest series in Japan, 27 (0.3%) were initially diagnosed as ER-/PgR+ breast carcinomas and we re-evaluated by IHC. RESULTS: The re-evaluated IHC showed that of the 27 patients with the initial results of ER-/PgR+, 12 were ER+/PgR+, 8 were ER-/PgR-, and 7 were ER-/PgR+. ER was negative in 12 of 27 patients (44.4%), and PgR was positive in 8 of 27 patients (29.6%). In our seven re-evaluated and confirmed as ER-/PgR+ cases, the staining proportions of tumor cells were 0% in ER and 1-69% (average 15.8%) in PgR. The average staining proportion of PgR in the re-evaluated ER-/PgR+ phenotype was lower than the initial diagnosis. Histological grading was as follows: grade I, one case; grade II, two cases; grade III, four cases. There were two lymph-node-positive cases. CONCLUSIONS: The ER-/PgR+ phenotype was confirmed after re-evaluation of ER and PgR assessment by a different pathologist. We recommend that pathologists discuss with clinicians, or re-test and re-evaluate ER/PgR expression, particularly in low-grade carcinoma and with a high staining proportion of PgR in the ER-/PgR+ phenotype.

Inhibitory effects of the cyclooxygenase-2 inhibitor, etodolac, on colitis-associated tumorigenesis in p53-deficient mice treated with dextran sulfate sodium.

Non-steroidal anti-inflammatory drugs (NSAIDs) and cyclooxygenase-2 (COX-2) inhibitors are representative agents for the chemoprevention of sporadic colorectal neoplasia. However, few reports have described the chemopreventive effects of such agents on colitis-associated tumorigenesis. To clarify whether treatment with the COX-2 inhibitor may reduce the risk of colitis-associated neoplasia, we investigated the effect of one such agent, etodolac, on tumorigenesis in the colitis-associated neoplasia model using p53-deficient mice treated with dextran sulfate sodium (DSS). The p53-/- mice were divided into four groups: i) treatment with DSS + etodolac, then after two cycles of DSS, the mice were given distilled water for 84 days. In addition, etodolac was administered three times a week at a dose of 10 mg/kg body weight throughout the experiment. ii) Treatment with two cycles of DSS only, followed by distilled water for 84 days. iii) Treatment with etodolac alone. iv) Distilled water alone was administered to the control group. The incidence of mice with neoplasia was 82.4% in the DSS + etodolac group and 100% in the DSS-alone group. No neoplasia was observed in the etodolac-alone and control groups. The mean (+/- SEM) number of total neoplasias per mouse was 1.29+/-0.2 in the DSS + etodolac group and 3.0+/-0.52 in the DSS-alone group, the inter-group difference being significant (p<0.01). There was no significant difference in the inflammation score between these two groups. These results showed that treatment with etodolac significantly reduced the occurrence of neoplasia, suggesting that this COX-2 inhibitor has chemopreventive activity against colitis-associated tumorigenesis.

Preoperative globulin-to-albumin ratio predicts outcome after curative resection in patients with gastric cancer.

AIM: The globulin-to-albumin ratio (GAR) is useful for prognostication of patients with various cancers. However, the significance of GAR in gastric cancer (GC) remains unclear. Our purpose was to investigate the relationship between the GAR and outcome after curative resection in GC patients. METHODS: Three-hundred and seventy-six patients who had undergone curative resection for GC were retrospectively reviewed. Univariate and multivariate analyses using the Cox proportional hazard model were performed to detect clinical characteristics that correlated with overall survival (OS), and their cut-off values were identified using receiver operating characteristic (ROC) curve analyses. Kaplan-Meier analysis and log-rank test were used for comparison of OS and relapse-free survival (RFS). RESULTS: Multivariate analysis using 17 clinical characteristics selected by univariate analyses revealed that GAR (>0.80/≤0.80) was significantly associated with OS (hazard ratio [HR], 2.305; 95% CI, 1.122-4.735; P = 0.023), as well as lymph node metastasis (presence/absence) (HR, 2.417; 95% CI, 1.077-5.426; P = 0.032), neutrophil-to-lymphocyte ratio (>2.7/≤2.7) (HR, 2.368; 95% CI, 1.138-4.930; P = 0.002), and serosal invasion (presence/absence) (HR, 3.443; 95% CI, 1.048-11.31; P = 0.042). Kaplan-Meier analysis and log-rank test demonstrated that the OS and RFS of patients with a high GAR (>0.80) were significantly poorer than those with low GAR (≤0.80). CONCLUSIONS: The GAR is a useful predictor of postoperative outcome among GC patients undergoing curative resection.

Rectal angiolipoma diagnosed after surgical resection: a case report.

Angiolipoma is a common benign tumor with a characteristic vascular component that usually occurs in subcutaneous tissue. Although lipomas are frequently encountered at colonoscopy as submucosal tumors, angiolipomas are rarely found in the gastrointestinal tract including the large intestine. Here we report a 77-year old Japanese man who underwent transanal resection of a tumor that was diagnosed tentatively as a leiomyoma. Histologically, the tumor consisted of mature fat cells and blood vessels. Immunohistochemically, the tumor cells were negative for c-kit and HMB-45, which are consistently expressed in tumors such as gastrointestinal stromal tumor and angiomyolipoma. The tumor was therefore diagnosed as an angiolipoma. This is thought to be the first case report of a surgically resected angiolipoma of the rectum.

Melting Temperature Mapping Method: A Novel Method for Rapid Identification of Unknown Pathogenic Microorganisms within Three Hours of Sample Collection.

Acquiring the earliest possible identification of pathogenic microorganisms is critical for selecting the appropriate antimicrobial therapy in infected patients. We herein report the novel "melting temperature (Tm) mapping method" for rapidly identifying the dominant bacteria in a clinical sample from sterile sites. Employing only seven primer sets, more than 100 bacterial species can be identified. In particular, using the Difference Value, it is possible to identify samples suitable for Tm mapping identification. Moreover, this method can be used to rapidly diagnose the absence of bacteria in clinical samples. We tested the Tm mapping method using 200 whole blood samples obtained from patients with suspected sepsis, 85% (171/200) of which matched the culture results based on the detection level. A total of 130 samples were negative according to the Tm mapping method, 98% (128/130) of which were also negative based on the culture method. Meanwhile, 70 samples were positive according to the Tm mapping method, and of the 59 suitable for identification, 100% (59/59) exhibited a "match" or "broad match" with the culture or sequencing results. These findings were obtained within three hours of whole blood collection. The Tm mapping method is therefore useful for identifying infectious diseases requiring prompt treatment.

Troglitazone suppresses cell growth of KU812 cells independently of PPARgamma.

We examined the effects of troglitazone, one of thiazolidinedione derivatives on human basophilic leukemia cell line KU812. Troglitazone caused the suppression of cell growth, which was suggested to result from the decrease in cyclin E and the hyperphosphorylated form of retinoblastoma tumor suppressor gene product (pRb). In addition, troglitazone caused a decrease in histamine secretion due to the reduced expression of histidine decarboxylase mRNA. Peroxisome proliferator-activated receptor (PPAR)-gamma mRNA was undetectable by reverse transcription-polymerase chain reaction (RT-PCR) in KU812 cells. These findings suggested that troglitazone suppressed cell growth and histamine synthesis independently of PPARgamma.

Prognostic nutritional index is associated with survival after total gastrectomy for patients with gastric cancer.

AIM: To investigate the influence of clinical characteristics including nutritional markers on postoperative survival in patients undergoing total gastrectomy (TG) for gastric cancer (GC). PATIENTS AND METHODS: One hundred fifty-four patients were enrolled. Uni- and multivariate analyses using the Cox proportional hazard model were performed to explore the most valuable clinical characteristic that was associated with postoperative survival. RESULTS: Multivariate analysis using twelve clinical characteristics selected from univariate analyses revealed that age (≤ 72/>72), carcinoembryonic antigen (≤ 20/>20) (ng/ml), white blood cell count (≤ 9.5/>9.5) (× 10(3)/mm(3)), prognostic nutritional index (PNI) (≤ 45/>45) and lymph node metastasis (negative/positive) were associated with postoperative survival. Kaplan-Meier analysis and log-rank test showed that patients with higher PNI (>45) had a higher postoperative survival rate than those with lower PNI (≤ 45) (p<0.001). CONCLUSION: PNI is associated with postoperative survival of patients undergoing TG for GC and is able to divide such patients into two independent groups before surgery.

Clinical significance of an inflammation-based prognostic system for gastric cancer patients with a preoperative normal serum level of carcinoembryonic antigen.

AIM: To investigate the significance of the Glasgow Prognostic Score (GPS) for predicting the postoperative survival of gastric cancer (GC) patients with a normal preoperative serum level of carcinoembryonic antigen (CEA). Because CEA is a useful marker for prognostication of several types of cancer, some patients with GC have a normal CEA level. On the other hand, the GPS has been established as a valuable inflammation-based prognostic system for cancer patients. PATIENTS AND METHODS: Among 650 patients who had undergone elective surgery for GC, 425 with a normal preoperative serum CEA level (≤5.0 ng/ml) were enrolled. Uni- and multivariate analyses were performed to evaluate the relationship of the GPS to overall survival. The Kaplan-Meier analysis and log rank test were used to compare the survival curves among patients with GPS 0, 1 and 2. RESULTS: Multivariate analysis using clinical characteristics selected from univariate analyses revealed that the GPS (0, 1/2) was associated with overall survival (hazard ratio=2.048; 95% C.I. (confidence interval)=1.002-4.185; p=0.049) along with age (≤70/>70) (years), sex, tumor type (3, 4, 5/0, 1, 2), lymph node metastasis (presence/absence) and platelet count (≤35/>35) (×10(4)/mm(3)). The Kaplan-Meier analysis and log rank test demonstrated that there were significant differences in overall survival among patients with GPS 0, 1 and 2 (p<0.001). CONCLUSION: Even if GC patients have a normal serum level of CEA, the GPS is able to predict their postoperative survival and classify such patients into three independent groups before surgery.

Needlescopic surgery for liver, gallbladder and spleen diseases.

BACKGROUND: We propose that needlescopic surgery (NS) should be considered as a way of improving the esthetic result and post-operative quality of life of patients and of reducing costs and stress on surgeons, and we have evaluated the results of NS. METHODS: We used NS in 157 patients between May 1998 and December 2010: cholecystectomy in 150 patients, marsupialization of splenic and hepatic cysts in 4 and splenectomy in 3, respectively. Under general anesthesia, one 12-mm and two or three 2- or 3-mm ports were introduced into the operative field. The specimen was retrieved from the 12-mm wound using a plastic bag. RESULTS: The procedures were successfully completed in all patients without conversion to an open procedure. In eight (5.3%) of 150 cholecystectomies a change to 5-mm instruments was required. The mean operation times and postoperative hospital stays for cholecystectomy, splenectomy, and marsupialization of splenic and hepatic cysts were 80.2 min and 3.2 days, 167 min and 5.6 days, 170 min and 7 days, and 120 min and 7 days, respectively. There were a few perioperative complications. The most important factor for reducing operation time and achieving a low conversion rate is the use of at least one 3- or 5-mm port for the grasping instruments in cholecystectomy. We recognized a residual cyst requiring splenectomy 62 months after marsupialization in one case. Technical points for performing safe procedures on solid organs were: no direct organ mobilization to avoid organ injuries, the rotation of the operating table and the utilization of organ gravity to create a better operative field, the minimum use of the needlescope to perform a safe maneuver and the improvement of bi-manual technique. CONCLUSIONS: NS is a safe and feasible procedure for achieving minimal invasive surgery. We should consider NS as a first choice to treat operable diseases in this laparoscopic era.

Intraoperative exploration of biliary anatomy using fluorescence imaging of indocyanine green in experimental and clinical cholecystectomies.

BACKGROUND/PURPOSE: We evaluated the usefulness of intraoperative exploration of the biliary anatomy using fluorescence imaging with indocyanine green (ICG) in experimental and clinical cholecystectomies. METHODS: The experimental study was done using two 40-kg pigs and the clinical study was done in 12 patients for whom cholecystectomy was planned from January 2009 to June 2009. Initially we used a laparoscopic approach for the evaluation of fluorescence imaging of the biliary system in the two pigs. Then the clinical study was started on the basis of these experimental results. ICG (1.0 ml/body of 2.5 mg/ml ICG) was infused 1-2 h before surgery. With the subjects under general anesthesia we observed in real time the condition of the biliary tract under the guidance of fluorescence imaging employing an infrared camera or a prototype laparoscope. ICG was added intravenously to observe the location or flow condition of the cystic artery. RESULTS: We obtained a clear view of the biliary tract and the location of the cystic duct in the two pigs. Local compression with a transparent hemispherical plastic device was effective for offering a clearer view. The biliary tract, except for the gallbladder, was clearly recognized in all clinical subjects. Local compression with a transparent hemispherical plastic device for open cholecystectomy and a flat plastic device for laparoscopy provided clearer visualization of the confluence between the cystic duct and common bile duct or common hepatic duct. The location of the cystic artery was revealed after division of the connective tissues, and the flow condition of the cystic artery was confirmed 7-10 s after intravenous re-infusion of ICG. There were no adverse events related to the intraoperative procedure or the ICG itself. CONCLUSIONS: This method is safe and easy for the identification of the biliary anatomy, without requiring cannulation into the cystic duct, X-ray equipment, or the use of radioactive materials. Although fluorescence imaging is still at an early stage of application in comparison with ordinary intraoperative cholangiography, we expect that this method will become routine, offering a lower degree of invasiveness that will help avoid bile duct injury.