Effects of aging on laboratory animals: I. Light and scanning electron microscopy of dog gallbladder

This study was begun in order to illustrate the changes that dog gallbladder undergo with age. Young (<8 weeks) and old (> 12 years) dogs were anesthetized, perfusion fixed and prepared for light and scanning electron microscopy. Observations with the light microscope suggested that gallbladder mucosa of young dogs was more folded and had few Rokitansky-Aschoff sinuses while the aging canine gallbladder mucosa appeared flattened and cystic. In addition, the gall-bladder lamina propria of old dogs exhibited increased numbers of lymphocytes and plasma cells, a less conspicuous vascular network, and lymph nodules. Few differences were noted in the muscular layer, the perimuscular connective tissue layer, and in the serosal layer between the young and old dogs. Scanning electron microscope observations of gallbladder mucosa tended to support the light microscopic data. The puppy gallbladder mucosa showed more folds and raised cell surfaces with the typical hobnail appearance while the aging canine gallbladder mucosa had few folds and considerable cellular flattening. These morphological findings suggest a reduced surface for absorption in the aging gallbladder.     

Lymphangiogenic and angiogenic microvessel density in human primary sporadic colorectal carcinoma

AIM： To investigate the distribution pattern of lymphatic vessels and microvessels in sporadic colorectal carcinoma （SCRC） and their relationship to metastasis and prognosis. METHODS： The lymphatic vessel density （LVD） and microvessel density （MVD） in tumor tissue obtained from 132 patients with primary SCRC, including 74 with metastases and 58 without metastases, were evaluated by immunohistochemistry using antibodies directed against D2-40 and yon Willebrand factor （vWF）. RESULTS： （1） The lymphatic vessels and microvessels at central portions of SCRC often had a reticular architecture with numerous tiny and ill-defined lumina, while those at tumor borders had large and open lumina. The LVD and MVD were both obviously higher in colorectal cancer patients with metastases than in those without （P 〈 0.001）. （2） For each one lymphatic vessel increased, there was a 1.45-fold increase in the risk of metastasis in SCRC. The specificity and sensitivity of LVD in predicting metastasis or non-metastasis in SCRC were 71.62% and 56.90%, respectively, and the corresponding LVD was 5. For each one microvessel increased, there was a 1.11-fold increase in the risk of metastasis in SCRC. The specificity and sensitivity of MVD were 66.22% and 51.72%, respectively. （3） Double labeling immunohistochemistry showed D2-40 immunoreactivity to be specific for lymphatic vessels. （4） Univariate analysis indicated that high LVD, high MVD, as well as co-accounting of high LVD and high MVD were associated with patient＇s poor disease-free survival （Puni 〈 0.05）; multivariate analysis indicated that co-accounting of LVD and MVD was an independent prognostic factor of colorectal cancer, CONCLUSION： D2-40 is a new specific antibody for lymphatic endothelial cells. Lymphogenesis and angiogenesis are commonly seen in SCRC, especially at tumor borders. The detection of LVD and MVD at tumor borders may be useful in predicting metastasis and prognosis in patients with SCRC, and, in particular, coa     

Expression profiling suggests a regulatory role of gallbladder in lipid homeostasis

AIM:To examine expression profile of gallbladder using microarray and to investigate the role of gallbladder in lipid homeostasis. METHODS: 33P-labelled cDNA derived from total RNA of gallbladder tissue was hybridized to a cDNA array representing 17 000 cDNA clusters. Genes with intensities ≥2 and variation <0.33 between two samples were considered as positive signals with subtraction of background chosen from an area where no cDNA was spotted. The average gray level of two gallbladders was adopted to analyze its bioinformatics. Identified target genes were confirmed by touch-down polymerase chain reaction and sequencing. RESULTS: A total of 11 047 genes expressed in normal gallbladder, which was more than that predicted by another author, and the first 10 genes highly expressed (high gray level in hybridization image), e.g. ARPC5 (2 225.88±90.46), LOC55972 (2 220.32±446.51) and SLC20A2 (1 865.21±98.02), were related to the function of smooth muscle contraction and material transport. Meanwhile, 149 lipid-related genes were expressed in the gallbladder, 89 of which were first identified (with gray level in hybridization image), e.g. FASN (11.42±2.62), APOD (92.61±8.90) and CYP21A2 (246.11±42.36), and they were involved in each step of lipid metabolism pathway. In addition, 19 of those 149 genes were gallstone candidate susceptibility genes (with gray level in hybridization image), e.g. HMGCR (10.98±0.31), NPC1 (34.88±12.12) and NR1H4 (16.8±0.65), which were previously thought to be expressed in the liver and/or intestine tissue only. CONCLUSION: Gallbladder expresses 11 047 genes and takes part in lipid homeostasis.     

Effect of rhubarb on contractile response of gallbladder smooth muscle strips isolated from guinea pigs

AIM: To investigate the effect of rhubarb on contractile response of isolated gallbladder muscle strips from guinea pigs and its mechanism. METHODS: Guinea pigs were killed to remove the whole gallbladder. Two or three smooth muscle strips (8 mm×3 mm) were cut along the longitudinal direction. The mucosa on each strip was carefully removed. Each longitudinal muscle strip was suspended in a tissue chamber containing 5 ml Krebs solution (37℃), bubbled continuously with 950 mL/L O2and 50 mL/L CO2. The resting tension (g), mean contractile amplitude (mm), and contractile frequency (waves/min) were simultaneously recorded on recorders. After 2-h equilibration, rhubarb (10, 20, 70, 200, 700, 1 000 g/L) was added cumulatively to the tissue chamber in turns every 2 min to observe their effects on gallbladder. Antagonists were given 3 min before administration of rhubarb to investigate the possible mechanism. RESULTS: Rhubarb increased the resting tension (from 0 to 0.40±0.02, P<0.001), and decreased the mean contractile amplitude (from 5.22±0.71 to 2.73±0.41, P<0.001). It also increased the contractile frequency of the gallbladder muscle strips in guinea pigs (from 4.09±0.46 to 6.08±0.35, P<0.001). The stimulation of rhubarb on the resting tension decreased from 3.98±0.22 to 1.58±0.12 by atropine (P<0.001), from3.98±0.22 to 2.09±0.19 by verapamil (P<0.001) and from 3.98±0.22 to 2.67±0.43 by phentolamine (P<0.005). But the effect was not inhibited by hexamethonium (P>0.05). In addition, the action of mean amplitude and frequency was not inhibited by the above antagonists. CONCLUSION: Rhubarb can stimulate the motility of isolated gallbladder muscle strips from guinea pigs. The stimulation of rhubarb might be relevant with M receptor, Ca2+ channel and α receptor partly.     

The role of prostaglandins E and F in acalculous gallbladder disease

Prostaglandins have been postulated to be involved in the formation of gallstones and the pain and inflammation of calculous gallbladder disease. This report evaluated prostaglandin E and F levels in patients with acalculous gallbladder disease. Control gallbladders were obtained from patients undergoing cholecystectomy during insertion of hepatic artery catheters for regional, hepatic chemotherapy. Patients without gallstones and with long-standing post-prandial biliary colic with abnormal cholecystokinin administration underwent cholecystectomy for chronic acalculous cholecystitis. A third group of patients underwent cholecystectomy for acute acalculous cholecystitis. Gallbladder mucosa and muscle were separated, and prostaglandin E and F concentrations in mucosal and muscle or mucosa were identified in gallbladders from patients with chronic acalculous cholecystitis compared to gallbladders from patients without biliary tract symptoms. In gallbladders from patients with acute acalculous cholecystitis a seven-fold increase in PGE production by muscle tissue and mucosal cells was found. The more histologically inflamed gallbladders had higher mucosal and muscle prostaglandin E concentrations than were found in less inflamed gallbladders. Prostaglandin F levels were not significantly changed or were decreased, resulting in a significant increase in the ratio of PGE/PGF in acutely diseased gallbladders when compared to normal gallbladders. Prostaglandin E may be a manipulatable intermediary in the sequence of events that results in the development of acute acalculous cholecystitis.     

Effect of Fructus Psoraleae on motility of gallbladder isolated smooth muscle strips from guinea pigs

AIM: To observe the effect of Fructus Psoraleae on motility of isolated gallbladder muscle strips of guinea pigs and its mechanism. METHODS: Guinea pigs were hit to lose consciousness and the whole gallbladder was removed quickly. Two or three smooth muscle strips (8 mm×3 mm) were cut along a longitudinal direction. The mucosa was gently removed. Every longitudinal muscle strip was suspended in a tissue chamber which was continuously perfused with 5 mL Krebs solution (37℃), pH 7.4, and aerated with 950 mL/L O2 and 50 mL /L CO2. The isometric response was recorded with an ink-writing recorder. After 2 h equilibration under 1 g-load, 50μL Fructus Psoraleae (10, 20, 70, 200, 700, 1000 g/L) was added cumulatively into the tissue chamber in turn every 2 min to observe their effects on gallbladder muscle strips (cumulating final concentration of Fructus Psoraleae was 0.1, 0.3, 1.0, 3.0, 10.0, 20.0 g/L). The antagonists, including 4-DAMP, benzhydramine, hexamethonium, phentolamine, verapamil and idomethine were given 2 min before Fructus Psoraleae respectively to investigate the mechanisms involved. RESULTS: Fructus Psoraleae dose-dependently increased the resting tension (r=0.992, P<0.001), decreased the mean contractile amplitude (r=0.970, P<0.001) and meanwhile increased the contractile frequency of the gallbladder muscle strip in vitro (r=0.965, P<0.001). The exciting action of Fructus Psoraleae on the resting tension could be partially blocked by 4-DAMP (the resting tension decreased from 1.37±0.41 to 0.70±0.35, P<0.001), benzhydramine (from 1.37±0.41 to 0.45±0.38, P<0.001), hexamethonium (from 1.37±0.41 to 0.94±0.23, P<0.05), phentolamine (from 1.37±0.41 to 0.89±0.22, P<0.01) and verapamil (from 1.37±0.41 to 0.94±0.26, P<0.05). But the above antagonists had no significant effect on the action of Fructus Psoraleae-induced mean contractile amplitude (P>0.05). Moreover, the increase of the contractile frequency due to Fructus Psoraleae was inhibited by 4-DAMP (decreased from 8.3±1.2 to 6.8±0.5, P<0.01) and hexamethonium (from 8.3±1.2 to 7.0±0.9, P<0.05).Idomethine had no significant effect on the Fructus Psoraleaeinduced responses (P>0.05). CONCLUSION: Fructus Psoraleae enhances the motility of isolated gallbladder muscle strips from guinea pigs, in a dose-dependent manner. The effect of Fructus Psoraleae is partly related to M3, N receptor, a receptor, H1 receptor, Ca2 channel, but not related to prostaglandin.     

H pylori exist in the gallbladder mucosa of patients with chronic cholecystitis

AIM:To study whether H pylori locate in the gallbladder mucosa of patients with chronic cholecystitis.METHODS:Using Warthy-Starry(W-S)silver stain and immunohistochemistry stain with anti-H pylori antibodies,we screened paraffin specimens in 524 cases of cholecystitis.H pylori urease gene A(HPUA)and H pylori urease gene B(HPUB)were analyzed by polymerase chain reaction(PCR)in the fresh tissue specimens from 81 cases of cholecystitis.RESULTS:H pylori-like bacteria were found in 13.55% of the gallbladders of the cholecystitis patients using W-S stain.Meanwhile,bacteria positive for H pylori antibodies were also found in 7.1% of the gallbladders of patients with cholecystitis by immunohistochemistry.Of 81 gallbladders,11 were positive for both HPUA and HPUB,4 were positive for HPUA only and 7 were positive for HPUB only.CONCLUSION:H pylori exist in the gallbladders of patients with chronic cholecystitis.     

Human greater and canine lateral saphenous veins. A morphologic and pharmacologic study.

Canine lateral saphenous vein and human greater saphenous vein were studied together to detmine similarities and differences. Helical strips of both vessels contracted to norepinephrine with similar sensitivity. Canine veins developed more isometric tension than human veins treated similarly. Histological studies show canine veins to have little connective tissue and a single circular layer of smooth muscle. Human veins have much connective tissue and three layers of smooth muscle, an inner circular layer, middle longitudinal layer and outer circular layer.     

How lymph is drained away from the human papillary muscle: anatomical conditions.

The lymphatic and venous bed was evaluated in 42 human heart papillary muscles. The lymphatics form networks in the superficial and deep layers of the subendocardium and in the myocardium of the papillary muscles. In the apical area, the subendocardial lymphatics gradually merge with the fine myocardial, developing arcades at the origins of chordae tendineae. Thicker myocardial lymphatics leave the area passing along the connective tissue septa, often in close vicinity to the blood vessels. The injection method failed to reveal deep lymphatics in the connective tissue of chordae tendineae. Lymphovenous anastomoses were disclosed in the papillary muscle. The possible morphological pathways for lymphatic drainage of the papillary muscle are as follows: (1) the lymphatic bed subendocardial and myocardial, joining on the ventricular wall bed at the muscle base, and (2) lymphovenous anastomoses, passing along from the myocardial and subendocardial lymphatic bed into the ventricular wall veins and the sinus coronarius system, or into the thebesian veins and further directly into the heart ventricles.     

Morphological changes of the hamster gallbladder in the contracted and distended states

The hamster gallbladders were investigated to clarify the morphological changes in the contracted and distended states. 1) The mucosal surface showed the highly convoluted folds in the contracted state, and the flat structure in distended state. 2) The height and width of mucosal epithelial cells were changed in correspondence with the contracted and distended state of the gallbladder. 3) A subepithelial blood capillary formed a well developed network, and possessed many pores opening mainly toward the mucosal epithelial cell. Three-dimensional structure of blood capillary was closely related to the morphological changes of the mucosal fold. 4) A contraction of distention of muscle layer of the gallbladder produced concomitant widening or narrowing of the lymphatic vessel lumen. It is suggested that muscular contraction worked as a "pump" for the lymphatic flow. 5) Smooth muscle cells in the muscle layer had many cell-to-cell junctions, but few neuromuscular junctions. It seems that there is a complicated interaction between nervous and humoral control mechanisms.     

Quantitative evaluation and qualitative results of surgical lymphovenous anastomosis in lower limb lymphedema

PURPOSE: Treatment of lymphedema (LE) includes complex decongestive physiotherapy (manual lymphatic drainage, bandaging, exercises, skin care, elastic stockings). Surgical therapy is rarely useful. However, lymphovenous anastomosis (LVA) is the most used surgery in LE. We have assessed LVA in lower limb LE. METHODS: Thirteen patients (5 women, 8 men) with primary (n=10) or secondary LE (n=3) were included. Primary LE started at a mean age (+/- SD) of 28.9 +/- 14.5 years. LE was located in left lower limb (n=7), right (n=4) or both (n=2). LVA was performed 7.1 +/- 4.9 years after the onset of LE by the same surgeon. Two to five lymphatic vessels were used for LVA. Assessment of LVA was based upon objective criteria (volumetry, erysipelas) and subjective criteria (global discomfort, heaviness, cutaneous tenderness, difficulties for doing significant effort or walking more than 1 km). Global assessement of LVA was collected for each patient. RESULTS: Before LVA, excess of volume (+/- SD) of LE was 1906 +/- 1277 ml or 28.5 +/- 18% in comparison with the controlateral limb. After LVA, excess of volume (+/- SD) remained stable with 1863 +/- 1468 ml or 24.4 +/- 18.9%. Volumetry was appreciated with a mean (+/- SD) follow-up of 52 +/- 3 months. Frequency of erysipelas was unchanged for the 6 patients with recurrent episodes. Only heaviness and cutaneous tenderness were significantly reduced after LVA. But global discomfort (+/- SD) decreased from 6.7 +/- 2.7 to 5 +/- 3.2 on visual analogic scale (NS). No differences were observed for significant effort or walking more than 1 km. Global assessment of LVA by the patient was very good (n=3), good (n=2), intermediate (n=5) and bad (n=3). CONCLUSION: LVA failed to improve the volume of lower limb LE and reduce the frequency of erysipelas. LVA improves few subjectives criteria but not global discomfort. Further studies are needed to evaluate LVA and to select patients to obtain best results.     

Vasa vasorum of subcutaneous veins and lymph vessels

After intraarterial ink-injection the vasa vasorum of veins and lymphatic vessels in adults, dead born children and fetuses were demonstrated micropreparatorily. In their walls the larger veins of the saphena system possess a capillary network corresponding to the connective tissue and muscle structures/patterns in the vessel wall. No vasa vasorum could be found in the subcutaneous veins less than 1 mm in diameter. The lymphatic vessels of the inner prefascial bundle show a one-dimensional capillary network in the outer layers of the wall. Like the vasa vasorum of the saphena system it develops in the second half of fetal life.     

乙型肝炎患者胆囊黏膜中病毒抗原表达分析

目的通过对慢性乙型肝炎患者的胆囊组织进行HBsAg、HBcAg的检测,了解乙型肝炎病毒在胆囊黏膜组织中的定位及分布状况以及对胆囊功能的影响,探讨慢性乙型肝炎患者胆囊病变与乙肝病毒（HBV）感染的关系.方法以鼠抗-HBs单克隆抗体、兔抗-HBc多克隆抗体,采用免疫组化S-P（SP）法,检测胆囊黏膜组织中的HBsAg和HBcAg.血清乙型肝炎病毒感染标志物阳性患者石蜡包埋胆囊标本29例为研究对象,血清乙型肝炎病毒感染标志物检测阴性患者的胆囊标本12例为对照组.乙肝病毒血清学检查用ELISA法检测.结果（1）乙型肝炎患者胆囊黏膜中存在HBsAg和HBcAg,29例患者胆囊黏膜组织中HBsAg的检出率为37.9%（11/29）,HBcAg的检出率为20.68%（6/29）.对照组HBsAg和HBcAg各有1例阳性,阳性率为8.3%（1/12）,HBsAg主要呈弥漫胞浆型分布,光镜下呈棕黄色颗粒沉积,主要分布在胆囊黏膜上皮细胞中,HBcAg的分布呈胞浆型和核型两种形态,见于腺上皮细胞和成纤维细胞中;（2）HBV病毒抗原阳性组和阴性组均呈慢性炎症性改变,胆囊黏膜组织的病理改变无明显差异;（3）胆囊息肉与HBV病毒感染的关系密切,肝癌患者的癌旁组织和胆囊黏膜中病毒抗原表达明显.结论乙型肝炎病毒感染者的胆囊黏膜中存在HBV病毒抗原,提示乙型肝炎病毒在肝外组织胆囊黏膜中也存在感染.     

Calcitonin gene-related peptide: an inhibitor of bullfrog (Rana catesbeiana) gallbladder contraction in vitro.

The presence of a calcitonin gene-related peptide (CGRP)-like material was demonstrated in the gallbladder of the bullfrog, Rana catesbeiana, using immunocytochemistry and confirmed by radioimmunoassay. An intense immunocytochemical reaction was observed in nerves located in the smooth muscle layers and associated with blood vessels. No immunoreactive nerve fibers were associated with ganglia, nor were immunoreactive cell bodies observed. Radioimmunoassay showed that 25.03 +/- 2.5 pmol/g tissue of CGRP-like material was present. In vitro tension studies using gallbladder strips showed that CGRP exerted an inhibitory effect on both acetylcholine- and cholecystokinin octapeptide-induced tension but had no effect on KCl-, norepinephrine-, or cerulein-induced tension. CGRP may act directly on the gallbladder smooth muscle to inhibit contraction.     

缬草提取物对胆囊结石治疗作用与肝、胆囊组织内TNF及IL—1水平的关系

目的：探索缬草提取物对胆囊结石的治疗作用与肝、肝囊组织内肿瘤坏死因子（TNF）及白细胞介素－1（IL－1）水平的关系。方法：大耳白种家兔，手术植入胆固醇结石。实验组喂缬草提取物，对照组灌喂等容、等渗蒸馏水溶液。8周后，手术取出胆囊结石称重，测量肝脏、胆囊组织中TNF、IL－1水平，胆囊壁做组织学。结果：实验组结石平均重量较对照组明显为轻（P＜0．01）；结石消失率达75％。实验组肝组织内TNF、IL－1水平与对照组比较无明显差异（P＞0．05）。胆囊组织内TNF、IL－1水平明显低于对照组（P＜0．01）。对照组的胆囊组织呈明显炎性改变，实验组的胆囊组织则基本正常。结论：缬草提取物对胆囊胆固醇结石有明显的治疗作用，其作用与囊组织内TNF及IL－1水平有关。     

食管鳞癌组织中VEGF-D表达及其与淋巴管密度和淋巴转移的关系

目的 研究食管鳞状细胞癌(ESCC)组织中血管内皮细胞生长因子D(VEGF-D)的表达情况及其与淋巴管密度(LVD)的关系,并探讨其与食管癌淋巴结转移的关系.方法 免疫组化SABC法检测55例手术切除的食管鳞状细胞癌组织、癌旁组织、正常组织中淋巴管密度和VEGF-D的表达水平,对结果进行统计分析.结果 (1)食管癌组织...     

Morphological features of lymphatic and mesothelial communications in the broad ligament of the pig

The broad ligament containing uterine, paraovarian, and oviduct lymphatics was examined in the pig in various phases of the estrous cycle using light, scanning and transmission electron microscopy. The architecture of these regions differed and was independent of the lymphangions of the precollector and collector lymphatic vessels. Lymphangions were separated from mesothelium by connective tissue and/or muscle layers; however, in the vicinity of the thin walled paraovarian sac, large lymphangions were often compressed between two epithelial layers. Numerous lymphatic lacunae were in direct contact with the peritoneal and paraovarian sac cavities. The mesothelial lining of the broad ligament and the external and internal epithelium of the pig paraovarian sac displayed two distinct cell types. Only smaller cuboidal cells with prominent microvilli extended above the lymphatic endothelium. The surfaces of these cells were discontinuous and showed: 1) lymphatic stomata, 2) small pores or fenestrae, 3) a superficial network of epithelial-free communications with underlying connective tissue to the paraovarian sac in the postovulatory period independent of the lymphatic vasculature, and 4) endothelial (instead of epithelial) cells with crevice-like discontinuities in large portions of the internal sac surface during the follicular phase of estrus. Numerous lymphatic stomata had orifices composed of flattened cuboidal cells while lymphatic endothelial cells were characterized by macula or zonula adherent connections formed within rims of various sizes (up to 50 microns in diameter). During estrus, there were circular (0.5-2.0 microns) and irregular (to 10 microns) interendothelial openings in stomatal orifices with migrating cells. These morphologic findings suggest that absorption and passage of fluid, particles and cells between cavities and the lymphatic lumen in areas of the paraovarian lymphatic plexus in the pig is feasible.     

Immunocytochemical Localization of Lymphatic and Venous Vessels in Colonic Polyps and Adenomas

Histopathological localization of lymphatic vessels has been hindered because of a lack of suitable immunocytochemical markers for lymphatic vessels. Using lymphatic vessels endothelial hyaluronan receptor-1 (LYVE-1) immunocytochemical staining, hyperplastic polyps, tubular adenomas to villous adenomas, were investigated for lymphatic vessels compared with immunostained blood vessels using factor-8. Four cases each of hyperplastic polyps, tubular adenomas to villous adenomas, were routinely fixed in formalin and embedded in paraffin and were immunostained using goat anti-LYVE-1 for lymphatic vessels and rabbit anti-factor-8 for blood vessels. In normal colon and hyperplastic polyps, slender lymphatic vessels were noted in muscularis mucosa, which spread into the base of colonic crypt, whereas round venous vessels, they extend into lamina propria. In tubular adenomas, small lymphatic and venous vessels were noted in broad fibrous stalks. In villous adenomas, smaller lymphatic and venous vessels were noted in fine intervillous stroma. In normal colon and hyperplastic polyps, slender, irregularly shaped lymphatic vessels were present in muscularis mucosa, spreading into the base of the colonic crypt. In tubular adenomas, small lymphatic and venous vessels were noted in fibrous stalks. In villous adenomas, smaller lymphatic and venous vessels were noted in intervillous stroma. There are no increased lymphatic and venous vessels in intermucosal stroma and stalks of adenomas compared with normal colon.     

Matrix Composition in Opossum Esophagus

The esophagus of mammalian species is organized into mucosa, connective tissue, and muscle, but little is known about the matrix of these layers. We studied by immunohistochemistry the distribution of collagens, fibronectin, versican, and elastin in the smooth muscle segment of the American opossum. Cryosections were exposed to specific antibodies and fluorescent-stained using conjugates of rhodamine or isothiocyanate. Staining was scored by two observers. We found that collagen I was prominent in the submucosa and in the muscular septa; collagen III formed fibrillar meshes in the lamina propria and the submucosa but was virtually absent from the epithelial and muscular layers; collagen IV was restricted to the base of the epithelium; collagen V, in contrast to collagen III, was prominent in epithelium and muscularis mucosae and sparse in muscular septa and submucosa. Fibronectin distribution followed collagen III; it formed layers in lamina propria and submucosa and strands in muscle septa and between individual muscle cells. Versican distribution followed collagen V; it was prominent in large muscle septa and formed thick sheets at the boundaries of submucosa/circular muscle and of circular/longitudinal muscle. We also determined the tissue contents of protein, hexuronic acid, and fibronectin. The mucosal layers exceeded the muscular layers in their content of hexuronic acid and fibronectin but not protein. We conclude that individual layers of the smooth muscle esophagus each have their own characteristic matrix. Lamina propria and submucosa are similar with regard to fiber orientation but lamina propria contains relatively more collagen III (small fibril) and submucosa comparatively more collagen I (large fibril). Nonfibrillar collagen V and versican are particularly prominent specifically on the boundaries between contracting muscle tissue and connective tissue framework.     

Dissection of the gallbladder from the liver bed during laparoscopic cholecystectomy for acute or subacute cholecystitis

Current literature suggests that early laparoscopic cholecystectomy (LC) for acute cholecystitis (AC) can be safely performed within 72 h of symptom onset. However, for various reasons, in clinical practice, fibrosed gallbladders are frequently encountered during early LC for AC. The subserosal layer of the gallbladder wall can be divided into an inner and an outer layer. The inner layer has an abundant vasculature and some fibrous tissue; it abuts the muscularis propria. The outer layer consists of abundant fat tissue; it abuts the serosa or the liver parenchyma. In both patients with AC and those without cholecystitis, dissection of the gallbladder in the outer layer facilitates removal of the gallbladder without tearing the gallbladder or injuring the liver parenchyma behind the liver bed. However, in patients with AC lasting 72 h or more, the subserosal layer becomes solid and thick due to inflammatory cell infiltration and fibrotic change. Thus, adequate dissection of the outer layer becomes difficult. However, there is a layer between the inner and outer layers that can be dissected bluntly and easily. Thus, we could dissect the gallbladder from its liver bed safely, surely, and quickly by using our original technique which was validated by histological examination.