Studies on Ectopic ACTH-Producing Tumors: III. An Ectopic Melanocyte-Stimulating Hormone: Gel Chromatographic Finding and Time-Lapse Color Change of Frog Skin

On isolation of the main melanocyte-stimulating factor fromthe metastatic liver tumor of islet cell carcinoma of the pancreas,it was found to be a peptide resembling human ß-MSHin the time-lapse skin darkening response of Xenopus frogs,but different in the volume of elution on Sephadex gel chromatography.     

Extraperitoneal laparoscopic retroperitoneal lymph node dissection in supine position after chemotherapy for advanced testicular carcinoma

AIM: To evaluate the feasibility and usefulness of extraperitoneal laparoscopic retroperitoneal lymph node dissection (RPLND) in the supine position after chemotherapy for advanced testicular carcinoma. METHODS: Three patients with advanced testicular cancer underwent chemotherapy. Although serum markers were decreased compared with the normal range, residual masses requiring surgical resection were recognized by computed tomography scanning. We applied extraperitoneal laparoscopic RPLND. The patients were placed in the supine position and the first trocar was inserted two finger widths medial to the anterior iliac spine. The retroperitoneal space was dilated using a preperitoneal distention balloon. Two more ports were inserted into the retroperitoneal space and surgery proceeded thereafter. RESULTS: The residual tumors were completely resected by laparoscopy. The procedure required 250-310 min and the bleeding volume was below 50 mL. Although the histopathological findings consisted only of necrosis in all of the patients, one patient recurred at the same place. CONCLUSIONS: Extraperitoneal laparoscopic RPLND in the supine position for residual tumors after chemotherapy is technically feasible and useful in terms of postoperative recovery. With regard to cancer control, further evaluation should be necessary.     

多壁碳纳米管致大鼠肺损伤效应的研究

[目的]探讨多壁碳纳米管（multi-wall carbon nanotubes,MWCNTs）对大鼠的肺损伤效应。[方法]Wistar大鼠160只,随机分成5组：生理盐水对照组、脱氧核苷酸钠盐（deoxyribonucleic acid sodium salt,DNA钠盐）组、DNA钠盐-MWCNTs 2.0、10.0、20.0mg/mL3组（低、中、高染毒组）,并用DNA钠盐提高MWCNTs的分散度。采用气管滴注的方法染毒,一次滴注量为每kg体重1.5mL,每天1次,连续滴注3d,分别于滴注结束后24h、7d、28d、90d时间段,每实验组各处死8只动物,测定肺灌洗液中总蛋白（TP）、乳酸脱氢酶（LDH）、谷胱甘肽（GSH）和超氧化物歧化酶（SOD）等细胞毒性及细胞氧化损伤指标及大鼠体重,并观察动物肺组织病理学改变情况。[结果]MWCNTs染毒结束后24h,各染毒组大鼠进食及饮水量均有所下降。染毒结束后7、28、90d时,大鼠体重与滴注前体重相比有不同程度的升高,但染毒剂量越高,体重增加幅度越小。中、高剂量染毒组灌洗液中TP和LDH含量随着染毒浓度的增加而升高;灌洗液中GSH和SOD有不同程度的下降,但随时间的延长有不同程度的升高。电镜下,MWCNTs染毒后7d可观察到大鼠肺组织细胞核膜发生肿胀,核固缩,单位膜结构不清楚;内质网扩张,线粒体发生肿胀,嵴断裂等病理改变。[结论]MWCNTs可引起肺组织损伤和氧化损伤,对大鼠具有肺毒性作用。     

Specific IgE and IgG1 Responses to Subtilisin Carlsberg (Alcalase) in Mice: Development of an Intratracheal Exposure Model

The primary objective of this study was to examine the feasibilityof using a mouse model to evaluate the immunogenicity of proteinsas a potential method to determine occupational exposure guidelines.Mice were intratracheally administered a benchmark protein allergen,subtilisin Carlsberg (Alcalase) in detergent matrix once a weekfor 4 to 6 weeks and specific IgE and IgG1 levels were determined.In all experiments, specific IgE levels were determined by usinga rat basophilic leukemia cell (RBL) release assay, while specificIgG1 was measured by an ELISA. A good correlation was observedbetween IgE titers determined by the RBL assay and rat passivecutaneous anaphylaxis assay. Intratracheal administration ofprotease with detergent matrix was found to result in significantIgE and IgG1 responses that were dose related. Detergent matrixwas found to enhance the Alcalase-specific IgE and IgG1 responsewhen administered by the intratracheal route. The IgG1 responsewas much more robust, easier to measure, and found to followthe IgE response. These results suggest that a mouse intratrachealmodel is a feasible approach to examining the immunogenic potencyof enzymes using specific IgE or IgG1 as the end points. Additionaldevelopment and validation of the mouse model with other typesof proteins will be pursued.     

Specific Antibody Responses to Subtilisin Carlsberg (Alcalase) in Mice: Development of an Intranasal Exposure Model

An intranasal (i.n.) dosing model was developed in mice as apotential alternative to more difficult, time-consuming, andcostly guinea pig intratracheal (GPIT) or mouse intratrachealmodels for assessment of the respiratory immunogenicity of detergentenzymes. Using a benchmark enzyme, Alcalase (protease subtilisinCarlsberg), studies were conducted to standardize the modelin terms of mouse strain, dosing and serum harvest regimen,and the primary immunoglobulin endpoint to use. The primaryassay endpoint selected was the enzyme-specific IgG1 titer determinedby an Alcalase-specific ELISA. This is not the primary allergenicantibody in mice (IgE is); however, IgG1 is coregulated withIgE via the IL-4/TH2 pathway and may have a role in mediatingallergic-type responses. BDF1 mice (C57B1/6 x DBA/2) were selectedas representative of high responder strains, with high responseassociated with the H-2^b (C57B1/6) parent. The dosing regimenused for most studies incorporated three i.n. exposures (Days1, 3, and 10) and bleeding of the animals on Day 15. The animalswere anesthetized and then immunized by allowing them to inhale5-µl aliquots of dosing solution into each nostril ateach immunization. Positioning of the animals with their headsdown (vs up) may have allowed more of the dosing solution toremain in the nasal region for a slightly longer period of time,but did not change the eventual GI tract migration and excretionof each dose. The presence of a detergent matrix in the enzymedosing solution enhanced the IgG1 response. Immunizing withenzyme plus detergent gave highly consistent dose-response curvesfor Alcalase when evaluated over many studies. An enzyme-specificallergic antibody (IgE) response was weak and inconsistent underthe dosing regimen used to generate the IgG1 response, but wasstronger with longer-term dosing, consistent with the delayin IgE vs IgG1 responses seen in some other studies. Using IgG1as a surrogate for allergic sensitization, we have preliminarydata showing similar differential potencies between Alcalaseand other test enzymes as detected in previous GPIT tests. Onthe basis of these data, we believe the i.n. immunization/IgG1response model is a robust technique that may be useful in determiningthe relative immunogenicities of detergent enzymes and otherproteins.     

Frameless Stereotactic Biopsy with Intraoperative Computed Tomography “Assessment of Efficacy and Real Target Registration Error”

Frameless stereotactic brain biopsy (FSB) with navigation system has been widely used. We reported preliminary experience of FSB with intraoperative computed tomography (iCT) and examined the usefulness of this novel adjuvant technique and real target registration error (rTRE) of FSB. The FSB with 5-aminolevulinic acid (5-ALA) and iCT was performed on 10 patients. The gadolinium-enhanced lesions on magnetic resonance image were defined as the biopsy target. In the procedure, iCTs were scanned twice, for autoregistration of the navigation system and for confirmation of the position of the actual inserted biopsy needle. The red fluorescence of the samples was observed under excitation with violet-blue light through a low-cut filter of neurosurgical microscope. The distance between the planned target and the tip of the biopsy needle in the image of iCT was calculated in a workstation for the assessment of rTRE. The median volume of the target was 12.13 mL (0.06-39.15 mL). We performed the surgical procedure in a prone position in four patients. None to faint 5-ALA-induced fluorescence was observed in six samples. There existed no sampling errors. The mean target distance between the planned and real targets of the mean rTRE of FSB was 2.7 ± 0.56 mm. The real TRE of FSB was first reported and was larger than the reported rTRE exactly calculated from the fiducial registration error. iCT guarantees accurate tumor sampling with autoregistration regardless of the surgical position and prevents inaccurate biopsy to occur even with ALA fluorescence assistance.