Subserosal Injection Research Articles

Subserosal uterine injection of blue dye for the identification of the sentinel node in patients with endometrial cancer: a feasibility study.

To define the detection rate, sensitivity, and negative predictive value (NPV) of the sentinel node technique in patients with endometrial cancer. Patients with endometrial cancer after informed consent underwent subserosal injection of blue dye during hysterectomy in a tertiary gynae/oncology department between 2010 and 2014. The procedure was performed in all cases by the same team including two gynae/oncologist consultants and one trainee. All relevant perioperative clinicopathological characteristics of the population were recorded prospectively. The identified sentinel nodes were removed separately and a completion bilateral pelvic lymphadenectomy followed in all cases. Simple statistics were used to calculate the sensitivity and NPV of the method on per patient basis. Fifty-four patients were included in this study. At least one sentinel node was mapped in 46 patients yielding a detection rate of 85.2%. Bilateral detection of sentinel nodes was accomplished in only 31 patients (57.4%). The mean number of sentinel nodes was 2.6 per patient and the commonest site of identification was the external iliac artery and vein area (66%). Six patients (11%) had a positive lymph node, and in five of them, this was the sentinel one yielding a sensitivity of 83.3% and an NPV of 97.5%. The overall detection rate improved significantly after the first 15 cases; however, this was not the case for the bilateral detection rate. Our study is in accordance with previous studies of sentinel node in endometrial cancer and further demonstrates and enhances the confidence in the technique. In the current era of an ongoing debate on whether a systematic lymphadenectomy in patients with endometrial cancer is still necessary, we believe that the sentinel node is an acceptable alternative and should be applied routinely in tertiary centres following a strict algorithm.

Comparative analysis of survival in colon cancer undergoing sentinel lymph node mapping vs conventional surgery based on number of positive lymph nodes.

e15154 Background: Unlike in breast cancer or melanoma, resection during sentinel lymph node mapping (SLNM) in colon cancer (CCa) includes regional lymphadenectomy including SLNs and non-SLNs. However, SLNM often identifies micrometastases that can be missed by conventional (Conv) surgery and pathologic examination. It is unknown whether this impacts survival or recurrence. Hence, a retrospective analysis was undertaken to study overall (OS) and disease -specific (DSS) survival between patients (pts) undergoing SLNM vs Conv surgery based on the number of +ve LNs. Methods: SLNM was done by subserosal injection with blue dye followed by segmental resection including regional lymphadenectomy. All SLNs were ultrastaged and other nodes were examined by conv. methods with H&E. Results: There were 309 pts in SLNM (GpA) vs 499 pts in Conv surgery (GpB); with average no. of lymph nodes (LNs) and +ve LNs 17.3/1.6 vs 14.4/2.49 respectively. For GpA, success rate was 99.6% and the average no of SLN was 3. Of the pts in GpA vs GpB, 1+ve LN were found in 38% vs 27%, 2+ve LNs in 10% vs 16%, and > 2 LNs in 53% vs 57%, respectively. Comparing 5 years OS between GpA vs GpB, for 1+ve LN was 62.8% vs 52.38%, for 2 +ve LNs 72.7% vs 48.65% and for > 2 +ve LNs 35% vs 33.33%, respectively. Similarly, DSS for 1 +veLN was 54.4% vs 47.6%, 2+ve LNs 40% vs 40.54% and > 2+ve LNs, 30.4% vs 25.76%, respectively (Table). Conclusions: Compared to Conv surgery, SLNM identified higher no. of LNs per pt with high success rate. Five-year OS and DSS also are better in SLNM vs Conv surgery for all +ve LN gps. Hence, SLNM in CCa may have prognostic value. A larger multicenter trial needs to be done to validate such data. [Table: see text]

Comparative analysis of survival in colon cancer undergoing sentinel lymph node mapping versus conventional surgery based on number of positive lymph nodes.

734 Background: Unlike in breast cancer or melanoma, resection during sentinel lymph node mapping (SLNM) in colon cancer (CCa) includes regional lymphadenectomy including SLNs and non SLNs. However, SLNM often identifies micrometastases which can be missed by conventional (Conv) surgery and pathologic examination. It is unknown whether this impacts survival or recurrence. Hence, a retrospective analysis was undertaken to study overall (OS) and disease -specific (DSS) survival between patients (pts) undergoing SLNM vs Conv surgery based on the number of +veLNs. Methods: SLNM was done by subserosal injection with blue dye followed by segmental resection including regional lymphadenectomy. All SLNs were ultrastaged and other nodes were examined by conv. methods with H&E. Results: There are 309 pts in SLNM (GpA) vs 499 pts in Conv surgery (GpB); with average no. of lymph nodes (LNs) and +ve LNs 17.3/1.6 vs 14.4/2.49 respectively. For GpA, success rate was 99.6% and the average no of SLN was 3. Of the pts in GpA vs GpB, 1+ve LN were found in 38% vs 27%, 2+ve LNs in 10% vs 16%, and > 2 LNs in 53% vs 57%, respectively. Comparing 5 years OS between GpA vs GpB, for 1+ve LN was 62.8% vs 52.38%, for 2 +ve LNs 72.7% vs 48.65% and for > 2 +ve LNs 35% vs 33.33%, respectively. Similarly, DSS for 1 +veLN was 54.4% vs 47.6%, 2+ve LNs 40% vs 40.54% and > 2+ve LNs, 30.4% vs 25.76%, respectively(Table1.). Conclusions: Compared to Conv surgery, SLNM identified higher no. of LNs per pt with high success rate. Five-year OS and DSS also are better in SLNM vs Conv surgery for all +ve LN gps. Hence, SLNM in CCa may have prognostic value. A larger multicenter trial needs to be done to validate such data. [Table: see text]

Application of lymph node labeling with carbon nanoparticles by preoperative endoscopic subserosal injection in laparoscopic radical gastrectomy

Objective: To evaluate the application value of carbon lymph node tracing technique by preoperative endoscopic subserosal injection in laparoscopic radical gastrectomy. Methods: From June 2013 to February 2015, seventy eight patients with gastric cancer were enrolled and randomly divided into trial group and control group. Subserosal injection of carbon nanoparticles around the tumor was performed by preoperative endoscopic subserosal injection one day before the operation in trial group, while the patients routinely underwent laparoscopic gastrectomy in control group. Results of harvested lymph nodes, postoperative complications were compared between the two groups. Carbon nanoparticle-related side effect was also evaluated. Results: The average number of harvested lymph node in trial group was significantly higher than that in control group (35.5±8.5 vs 29.5±6.5, P<0.05). The rate of overall black-dyed harvested lymph node was 74.7% (1 035/1 386) in trial group, the black-dyed lymph node rate in D1 lymph node was 80.1%, which was significantly higher than that in D2 lymph node (69.8%, χ2=19.38, P<0.01). When comparing the lymph node with and without black-dyed in trial group, the rate of metastasis lymph node was significantly higher in lymph node with black-dyed (17.3% vs 4.0%, χ2=38.67, P<0.01). There was no significant difference in postoperative complications rate between two group (trial group 10.2%; control group 12.8%, χ2=0.00, P>0.05), and no carbon nanoparticle-related side effect was observed. Conclusion: Given a higher harvested lymph node number and a similar rate of complications, preoperative endoscopic subserosal injection of carbon nanoparticles was safe and feasible.

Enhancement of Gastric Ulcer Healing and Angiogenesis by Hepatocyte Growth Factor Gene Mediated by AttenuatedSalmonellain Rats

The present study developed an oral hepatocyte growth factor (HGF) gene therapy strategy for gastric ulcers treatment. An attenuated Salmonella typhimurium that stably expressed high HGF (named as TPH) was constructed, and the antiulcerogenic effect of TPH was evaluated in a rat model of gastric ulcers that created by acetic acid subserosal injection. From day 5 after injection, TPH (1 × 109 cfu), vehicle (TP, 1 × 109 cfu), or sodium bicarbonate (model control) was administered orally every alternate day for three times. Then ulcer size was measured at day 21 after ulcer induction. The ulcer area in TPH-treated group was 10.56 ± 3.30 mm2, which was smaller when compared with those in the TP-treated and model control groups (43.47 ± 4.18 and 56.25 ± 6.38 mm2, respectively). A higher level of reepithelialization was found in TPH-treated group and the crawling length of gastric epithelial cells was significantly longer than in the other two groups (P < 0.05). The microvessel density in the ulcer granulation tissues of the TPH-treated rats was 39.9 vessels/mm2, which was greater than in the TP-treated and model control rats, with a significant statistical difference. These results suggest that TPH treatment significantly accelerates the healing of gastric ulcers via stimulating proliferation of gastric epithelial cells and enhancing angiogenesis on gastric ulcer site.

Abstract 835: Comparison of direct transplantation of colon cancer cells vs. tumor xenografts using orthotopic mouse model for colon cancer progression and metastasis

Abstract Orthotopic transplantation is the process of implantation of cancer cells or tumor xenograft tissues into the organ of origin of the cancer. This procedure enables cells to grow and proliferate in their natural microenvironment. The orthotopic transplantation model has been observed to be highly amenable to cancer cells and recapitulates the human disease with high fidelity. Orthotopic transplantation is very common in clinical cancer research using athymic nude mouse as a model system. A major advantage of the orthotopic transplantation model is its ability to study the development and progression of cancer metastasis. The identification and therapeutic validation of anti-metastatic agents is of urgent need since till date there are no effective anti-metastatic therapies available for colon cancer. The procedure of xenograft transplant is time consuming and costly because of the involvement of more number of mice. Popular method for orthotopic transplant is to grow a subcutaneous tumor injecting human cancer cells and then transplant a small piece of xenograft to the immune deficient mouse for clinical study. In this study, we directly injected the tumor cells below the sub-serosal layer of the caecum to develop colorectal cancer bypassing the formation of xenograft and its subsequent transplantation. We compared the effectiveness in tumor progression and metastasis between the cell transplant and xenograft implant to the caecum. GFP labelled HCT116 cells were used for this study to develop colorectal cancer. First, we injected 8 million cells subcutaneously to the athymic nude mice to generate xenograft for further transplant to the caecum. On another set of experimental animals, a total of 2 million cells were directly injected below the sub-serosal membrane to develop primary tumor on the colon. Whole body GFP imaging revealed that within a week of transplantation of cancer cells or xenografts, primary tumor starts to develop to the colon and metastasized within 4-5 weeks after implantation. In both the cases, we record a 100% of primary tumor formation with 50-60% liver and 75-80% lung metastasis. To confirm the metastasis in liver and lung we studied GAPDH expression on the tissues and conduct H&amp;E /immunofluorescence staining. Whole body or organ GFP imaging also reveals that pancreas, kidney and associated intestinal lymph glands are severely affected by both the orthotopic transplantation methods. In conclusion, direct sub-serosal injection of cells to develop colorectal cancer primary and metastatic deposits is cost effective and time saving and thus seems to potentially have an advantage over xenograft transplant method. Citation Format: Anathbandhu Chaudhuri, Liying Geng, Jing Wang. Comparison of direct transplantation of colon cancer cells vs. tumor xenografts using orthotopic mouse model for colon cancer progression and metastasis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 835.

A Technique for Vascular Control During Robotic-assisted Laparoscopic Myomectomy.

To show and describe a unique method for improved vascular control when performing a robotic myomectomy. A video showing skeletonization of the uterine vessels and uterine artery occlusion to maximize vascular control and minimize blood loss during robotic-assisted myomectomy (Canadian Task Force classification III). Myomectomy is a procedure often reserved for the treatment of symptomatic leiomyomas in women who desire fertility preservation. Although traditionally performed via laparotomy, laparoscopic and robotic-assisted approaches are increasing in popularity. One of the limitations of myomectomy is the potential for increased blood loss and surgical morbidity. Although studies suggest a decrease in blood loss with a robotic approach, the risk of requiring a blood transfusion remains. In this video, we show a technique for maximizing vascular control during myomectomy in a Jehovah's Witness. In addition to subserosal injection with a dilute solution of vasopressin, we used vascular clips and bulldog clamps to temporarily occlude the uterine arteries. The uterine vessels were skeletonized at the level of the cervix to provide access for placing the vascular clips. Additionally, a peritoneal window was created in the mesosalpinx, and bulldog clamps were available to transiently occlude the utero-ovarian vessels in case of increased blood loss. Temporary occlusion of the uterine vessels can be used during robotic-assisted myomectomy to provide improved vascular control. We were able to use this technique during robotic-assisted myomectomy in a Jehovah's Witness patient. This approach can be considered for selected patients by experienced laparoscopic surgeons to lower blood loss and potentially decrease the need for transfusion.

Application of subserosal injection of carbon nanoparticles via infusion needle to label lymph nodes in laparoscopic radical gastrectomy

To explore the feasibility of subserosal injection of carbon nanoparticle via venous infusion needle to label lymph node and its application value in laparoscopic radical gastrectomy. Forty patients with gastric cancer were randomly divided into two groups (carbon nanoparticle group and control group). Subserosal injection of carbon nanoparticle around the tumor was performed via venous infusion needle laparoscopically at the beginning of surgery in carbon nanoparticles group, while the patients routinely underwent laparoscopic radical gastrectomy in control group. Results of harvested lymph nodes were compared between the two groups. The perioperative complications and the side effect of carbon nanoparticle were also evaluated. The average number of harvested lymph node in carbon nanoparticle group (31.7±7.6) was significantly higher than that in control group (19.8±6.1, P<0.05). The proportion of harvested small node (< 5 mm) in carbon nanoparticles group(61.0%) was higher than that in control group(43.3%, P<0.01). The mean harvest time in carbon nanoparticle group [(23.5±4.8) min] was shorter than that in control group [(32.6±5.5) min, P<0.05]. The rate of black-dyed harvested lymph node was 61.9% and the metastasis rate of black-dyed lymph node was 23.0% in carbon nanoparticle group, which were significantly higher than those without black-dyed(6.2%, P<0.05) and those in control group (15.7%, P<0.05). The operative time and perioperative complications were not significantly different between the two groups, and no serious side effect caused by carbon nanoparticle was observed. Subserosal injection of carbon nanoparticle via venous infusion needle to label lymph nodes during laparoscopic radical gastrectomy is safe and feasible. It can increase the number of harvested lymph node, especially the small node.

Comparison of sizes of sentinel and non-sentinel lymph nodes in colorectal cancers (CRCa).

557 Background: Sentinel lymph node (SLN) mapping (M) in CRCa with ultrastaging of SLNs find more nodes per patient (Pt) and more metastasis in the SLNs than non-SLNs (NSLNs) compared to conventional methods. We hypothesized that SLNs may represent a subset of nodes smaller than those detected by routine pathological exam, and hence missed by conventional methods. Hence, we compared the sizes of SLNs and NSLNs in CRCa pts undergoing SLNM. Methods: 365 pts underwent subserosal peritumoral injections of blue dye for SLNM. Blue stained nodes were tagged as SLNs. SLNs were sectioned at 5 levels, 4 stained with H &amp; E, one with cytokeratin (CK-IHC). The non-SLNs were sectioned at one level and stained with H &amp; E. Greatest diameter of nodes for sizes were measured on glass slides or from gross examination. Results: Of the total of 5,601 LNs (Average 15.3/pt) included, 912 were SLNs and 4689 non-SLNs. Of the total lymph nodes removed, 78% were 5mm or less; 69% of positive LNs and 97% of negative LNs were less than 9mm (Table). On average, the size of SLN was bigger (5.7mm) than NSLN (4.0mm). Both positive and negative SLNs were also bigger than positive and negative NSLNs (Table). Conclusions: Our study confirms that on average, SLNs are bigger than NSLNs whether positive or negative or as a whole. This also highlights diligent efforts needed by the pathologists, as 77% of all lymph nodes were less than 5mm. [Table: see text]

Robotic Flourescent Sentinel Node Identification in Endometrial Cancer

The role of lymphatic mapping and sentinel lymph node identification in endometrial cancer is evolving. It has been shown that finding sentinel nodal metastases can help guide the need for adjuvant therapy without the associated morbidity of comprehensive pelvic and para-aortic lymphadenectomy. A number of different modalities of sentinel node identification have been used and, to date, there is no standard approach. Among these modalities are subserosal injection, hysteroscopic-guided injection, and more recently direct cervical injection.

Laparoscopic Sentinel Lymph Node Identification in Patients with Colon Carcinoma Using a Near-Infrared Dye: Description of a New Technique and Feasibility Study

After promising results were obtained from studies in large animals, a technique using indocyanine green (ICG) is being introduced for sentinel lymph node (SLN) biopsy in colon cancer patients. Colon cancer patients without clinical signs of metastatic disease, presenting at the VU University Medical Center (Amsterdam, The Netherlands) or Kennemer Gasthuis (Haarlem, The Netherlands), were asked to participate in the study. During laparoscopy, a subserosal injection of 2.5 mg of ICG diluted in 1 mL of 0.9% NaCl plus 2% human albumin was performed using a percutaneously inserted long rigid or flexible needle. After injection, a near-infrared laparoscope (Olympus Corp., Tokyo, Japan) was used for lymph flow and SLN visualization. The SLNs were laparoscopically harvested and analyzed by a senior pathologist using multisectioning and immunohistochemistry. Fourteen patients were included (six women, eight men), with a median age of 75.5 (interquartile range [IQR], 67.8-81.0) years and a median body mass index of 25.1 (IQR, 22.7-26.0) kg/m(2). Median tumor diameter was 4.5 (IQR, 3.4-7.0) cm. At least one SLN was identified in all patients, with a median number of 2.0 (IQR, 2.0-3.3) SLNs. The median time between injection and identification of the SLN was 15.0 (IQR, 13.3-29.3) minutes. Positioning of the needle tip into the subserosal layer was found to be more effective using the flexible needle. When this flexible needle was used, less spill of dye was observed. All SLNs were negative. We observed four false-negative nodes, all after using a rigid needle. None of the patients showed an adverse reaction to the ICG injection. Preliminary results of laparoscopic sentinel node identification using a near-infrared dye show this procedure is safe and feasible. It was possible to detect lymph nodes in all patients. Large tumor size, drainage to adjacent lymphatic vessels, and the use of a rigid needle might contribute to false-negative nodes.

Early Gastric Cancer: Feasibility of CT Lymphography with Ethiodized Oil for Sentinel Node Mapping

To evaluate the feasibility of CT lymphography with ethiodized oil for sentinel node mapping in porcine stomachs and in patients with early gastric cancer. Approval for the animal study was obtained from the authors' institutional animal use and care administrative advisory committee, the clinical study was approved by the institutional review board, and informed consent was obtained from each participant. Five pigs underwent CT lymphography 1 hour after gastric subserosal injection of 1 mL ethiodized oil and sentinel node mapping with injection of 1 mL methylene blue. Ethiodized oil retention nodes were identified on the radiographic images of the gastric specimen and removed for histopathologic examination. In addition, 10 patients with early gastric cancer underwent CT lymphography with peritumoral injection of 1 mL ethiodized oil, followed by sentinel basin extirpation with CT and routine nodal dissection. The removed sentinel basins were examined by radiography. Histopathologic examination was performed for dissected nodes, including sentinel nodes. In each of the five pigs, CT showed one perigastric ethiodized oil retention node. After harvesting the ethiodized oil retention node, blue-stained areas were identified in the five removed nodes and intranodal ethiodized oil was detected on histopathologic examination. In all 10 patients, CT lymphography with ethiodized oil successfully defined the sentinel basin with ethiodized oil retention nodes. CT lymphography revealed 20 ethiodized oil retention nodes. After basin extirpation, 28 and 46 nodes were detected on radiographic and histopathologic examinations. Histopathologic examination revealed that one patient had micrometastases at two sentinel nodes and another patient had isolated tumor cells at one sentinel node. No patient had metastasis in nonsentinel nodes. CT lymphography with ethiodized oil may be a feasible method for sentinel node mapping in patients with early gastric cancer.

Animal model for training in sentinel lymph node biopsy of the stomach through combined methods

PURPOSE: Create and validate a proposed animal model for training in sentinel lymph node biopsy of the stomach. METHODS: In thirty-two rabbits, through a laparotomy, they received a subserosal injection of 0.1 ml of phytate labeled with technetium-99m (0.2 mCi) in the anterior wall of the gastric corpus, followed by 0.2 ml of Blue Patent V® 2.5%, through the same puncture site. Suspicious lymph nodes were searched in vivo at five, ten and 20 minutes, both visually (Blue Patent stained lymph nodes) and with a manual gamma radiation detector (to detect suspected radioactive lymph nodes). After 20 minutes, was performed resection of these for further evaluation of radioactivity (ex vivo) and histological study. RESULTS: Lymph nodes were identified in 30 rabbits (Average of 2.2 lymph nodes per animal). Of the 90 suspected lymph nodes that occurred in the study, 70 cases (77.8%) were histologically confirmed for lymphoid tissue. Of these, the majority were located in the periesophageal region of the gastric fundus. The sample presented a mortality rate of 6.25% and nine complications related to the method, which interfered in the identification of the lymph nodes. CONCLUSION: The animal model for sentinel node biopsy in rabbit stomachs proved to be feasible, with low complexity and reproduced the difficulties encountered for gastric lymph node biopsy in humans, being adequate for surgical training.

Nanoparticle Migration and Delivery of Paclitaxel to Regional Lymph Nodes in a Large Animal Model

The aim of this study was to demonstrate feasibility of migration and in situ chemotherapy delivery to regional lymph nodes (LN) in a large animal model using an expansile polymer nanoparticle (eNP) delivery system. Dual-labeled 50-nm and 100-nm eNP were prepared by encapsulating an IR-813 near-infrared (NIR) fluorescent dye within coumarin-conjugated expansile polymer nanoparticles (NIR-C-eNP). NIR imaging and fluorescent microscopy were used to identify intralymphatic migration of NIR-nanoparticles to draining inguinal or mesenteric LN after injection in swine hind legs or intestine. Nanoparticle-mediated intranodal delivery of chemotherapy was subsequently assessed with Oregon Green paclitaxel-loaded NIR-eNP (NIR-OGpax-eNP). NIR imaging demonstrated direct lymphatic migration of 50-nm, but not 100-nm, NIR-C-eNP and NIR-OGpax-eNP to the draining regional LNs after intradermal injection in the hind leg or subserosal injection in intestine. Fluorescent microscopy demonstrated that IR-813 used for NIR real-time trafficking colocalized with both the coumarin-labeled polymer and paclitaxel chemotherapy and was identified within the subcapsular spaces of the draining LNs. These studies verify nodal migration of both nanoparticle and encapsulated payload, and confirm the feasibility of focusing chemotherapy delivery directly to regional nodes. Regionally-targeted intranodal chemotherapy can be delivered to draining LNs for both skin and solid organs using 50-nm paclitaxel-loaded eNP.

Colorectal cancer lymph node staining by activated carbon nanoparticles suspensionin vivoor methylene bluein vitro

To investigate whether activated carbon nanoparticles suspension (ACNS) or methylene blue (MB) can increase the detected number of lymph nodes in colorectal cancer. Sixty-seven of 72 colorectal cancer patients treated at our hospital fulfilled the inclusion criteria of the study which was conducted from December 2010 to February 2012. Seven patients refused to participate. Eventually, 60 patients were included, and randomly assigned to three groups (20 in each group): ACNS group (group A), MB group (group B) and non-stained conventional surgical group (group C). In group A, patients received subserosal injection of 1 mL ACNS in a 4-quadrant region around the mass. In group B, the main artery of specimen was identified and isolated after the specimen was removed, and 2 mL MB was slowly injected into the isolated, stretched and fixed vessel. In group C, no ACNS and MB were injected. All the mesentery lymph nodes were isolated and removed systematically by visually inspecting and palpating the adipose tissue. No difference was observed among the three groups in age, gender, tumor location, tumor diameter, T-stage, degree of differentiation, postoperative complications and peritoneal drainage retention time. The total number of detected lymph nodes was 535, 476 and 223 in the three groups, respectively. The mean number of detected lymph nodes per patient was significantly higher in group A than in group C (26.8 ± 8.4 vs 12.2 ± 3.2, P < 0.001). Similarly, there were significantly more lymph nodes detected in group B than in group C (23.8 ± 6.9 vs 12.2 ± 3.2, P < 0.001). However, there was no significant difference between group A and group B. There were 50, 46 and 32 metastatic lymph nodes dissected in 13 patients of group A, 10 patients of group B and 11 patients of group C, without significant differences among the three groups. Eleven of the 60 patients had insufficient number of detected lymph nodes (< 12). Only one patient with T(4a) rectal cancer had 10 lymph nodes detected in group B, the other 10 patients were all from group C. Based on the different diameter categories, the number of detected lymph nodes in groups A and B was significantly higher than in group C. However, there was no statistically significant difference between group A and group B. The metastatic lymph nodes were not significant different among the three groups. Similarly, tumor location, T stage and tumor differentiation did not affect the staining results. Body mass index was a minor influencing factor in the two different staining methods. The stained lymph nodes can easily be identified from the mesenteric adipose tissues, and the staining time for lymph nodes was not significantly different compared with unstained group. None of the patients in groups A and B had drug-related complications. Both activated carbon nanoparticles suspension in vivo and methylene blue in vitro can be used as tracers to increase the detected number of lymph nodes in colorectal cancer.

Sentinel lymph node biopsy in endometrial cancer: Meta-analysis of 26 studies

Objective The validity of the sentinel lymph node (SLN) procedure for the assessment of nodal status in patients with endometrial cancer is unclear. We aimed to assess the diagnostic performance of this procedure. Methods We searched the PubMed and Embase databases for studies published before June 1, 2011. Eligible studies had a sample size of at least 10 patients, and reported the detection rate and/or sensitivity of the SLN biopsy. Results We identified 26 eligible studies, which included 1101 SLN procedures. The overall weighted-mean number of harvested SLNs was 2.6. The detection rate and the sensitivity were 78% (95% confidence interval [CI] = 73%–84%) and 93% (95% CI = 87%–100%), respectively. Significant between-study heterogeneity was observed in the analysis of the detection rate ( I-squared statistic, 80%). The use of pericervical injection was correlated with the increase of the detection rate ( P = 0.031). The hysteroscopic injection technique was associated with the decrease of the detection rate ( P = 0.045) and the subserosal injection technique was associated with the decrease of the sensitivity ( P = 0.049), if they were not combined with other injection techniques. For the detection rate, significant small-study effects were noted ( P < 0.001). Conclusions Although SLN biopsy has shown good diagnostic performance in endometrial cancer, such performance should be interpreted with caution because of significant small study effects. Current evidence is not yet sufficient to establish the true performance of SLN biopsy in endometrial cancer.

Value of sentinel lymph node mapping using a blue dye-only method in gastric cancer: a single-center experience from North-East Hungary

Forty percent of patients with gastric cancer have unnecessarily extended lymph node dissections with higher rates of morbidity and mortality than those in non-extended procedures. Successful sentinel lymph node (SLN) mapping may help to reduce the number of extended lymphadenectomies. SLN mapping was investigated by a blue dye-only method in patients with gastric cancer. The first cohort of patients (n=16) were marked submucosally by an endoscopist and in the second cohort of patients (n=23) a subserosal injection was performed by the surgeon. Thirty-nine patients, all Caucasians, underwent gastric resection or total gastrectomy with SLN biopsy using patent blue-dye mapping and modified D2 lymphadenectomy. The mapping procedure and the lymphadenectomy were supervised by the same surgeon. A total of 770 lymph nodes were removed and examined. The mean number of blue nodes was 4.3 per patient. In 22/23 cases at least one SLN showed tumor involvement. The sensitivity of SLN mapping was 95.7%, the false-negative rate was 4.3%, and the specificity was 100%. The negative predictive value was 93.8% and the positive predictive value was 100%. In cases of T1 and T2 tumors the sensitivity was 100%. We found the two marking methods (submucosal vs. subserosal) to be equivalent and there was no side-effect of the blue-dye mapping. Our results suggest that SLN mapping with blue dye alone represents a safe procedure that seems to be adaptable for non-obese patients undergoing open surgery for gastric cancer in the Eastern European region. The procedure has high sensitivity and specificity, especially in cases of T1 and T2 tumors.

A New Concept for the Role of Ex vivo Sentinel Lymph Nodes in Node-Negative Colorectal Cancer

We recently introduced ex vivo, intra-arterial methylene blue injection as a simple method to improve the lymph node (LN) harvest in gastrointestinal cancer. We now combined it with a novel ex vivo sentinel lymph node (evSLN) mapping technique. evSLN mapping was performed by subserosal (n=20) or submucosal (n=30) India ink injection. Subsequently, methylene blue was injected intra-arterially to enhance visibility of all LNs to improve the overall LN harvest. Manual LN dissection was carried out after fixing overnight. evSLNs nodes were identified by detecting carbon particles during histological examination. In primary node-negative cases, all detected LNs were step sectioned and immunohistochemically stained for pan-cytokeratin. India ink injection was easy to perform. Methylene blue injection failed in 1 case. The mean lymph node harvest was 42±18 LNs, and the SLN detection rate was 78%. The sensitivity for detecting metastases was 75%. The mean SLN number was 3±1. LN metastases were found in 20 of 47 malignant cases (43%). Skip metastases occurred in 4 cases. Of these cases, 3 showed involvement of at least 1 entire LN. True upstaging (N0→N1mi) was found in 1 of 23 cases (4%) within a SLN after advanced evaluation. Combination of methylene blue technique and ex vivo sentinel mapping is feasible, easy to perform, and cost effective. It guarantees an optimal LN harvest and has the potential to heighten the sensitivity of metastasis detection.

Enhancement of Gastric Ulcer Healing and Angiogenesis by Cochinchina Momordica Seed Extract in Rats

Cochinchina momordica seed is the dried ripe seed of Momordica cochinchinensis, a perennial vine. The antiulcer effect of an extract from cochinchina momordica seeds (SK-MS10) was evaluated in a rat model of acetic acid-induced gastric ulcers. Gastric ulcers were produced by subserosal injection of acetic acid. SK-MS10 (200 mg/kg) or vehicle was administered orally once per day for 14 days after the acetic acid injection. The stomach was removed and the ulcer size measured at day 7 and 14 of the treatment. Expression of vascular endothelial growth factor (VEGF) was assessed by real-time RT-PCR and Western blot analysis. In addition, the microvasculature density (MVD) adjacent to the ulcer margin was examined by immunohistochemistry. The treatment with SK-MS10 for 7 and 14 days significantly accelerated ulcer healing and increased the expression of mRNA (at day 7) as well as VEGF protein (at day 14) compared to the vehicle-treated rats. The MVD for factor VIII was also higher in the SK-MS10 treatment group compared to the vehicle-treated rats; however, these differences were not statistically significant. These results suggest that SK-MS10 treatment accelerates the healing of gastric ulcers via upregulation of VEGF and angiogenesis in an acetic acid rat model.

Neurochemical and morphological phenotypes of vagal afferent neurons innervating the adult mouse jejunum

Whilst much is known about the function and influence of vagal afferents on the mammalian upper gastrointestinal tract, the phenotypes of the different types of vagal afferent neurons innervating the jejunum is unknown. We have previously shown that spinal afferents supplying the jejunum are predominantly medium-sized sensory neurons that express specific combinations of transient receptor potential vanilloid type 1 (TRPV1), neuronal nitric oxide synthase (NOS) and calcitonin-gene related peptide (CGRP) and that they lack binding for isolectin B4 (IB4). This study aimed to identify the chemical phenotypes and somal sizes of jejunal afferent neurons in the mouse vagal ganglion. Jejunal vagal afferents were identified by retrograde labelling with sub-serosal injections of cholera toxin B (CTB) into the jejunal wall and assessed for IB4-binding, TRPV1-, NOS- and CGRP-immunoreactivities using fluorescent microscopy. Almost all (99%) of CTB-labelled vagal afferent neurons were small- and medium-sized sensory cells. Most (81%) jejunal vagal afferents bound IB4 but fewer (32%) expressed TRPV1. A quarter (25%) of those that bound IB4 co-expressed TRPV1-immunoreactivity whilst 77% of TRPV1-expressing jejunal vagal afferent neurons bound IB4. NOS (0%) and CGRP (0%) expression was absent from all CTB-labelled cells examined. In conclusion, vagal afferents innervating the jejunum differ in their expression of IB4, TRPV1, CGRP and NOS from their spinal counterparts, suggesting that the peripheral endings for extrinsic sensory neurons terminating within the enteric nervous system can be identified selectively.