Introduction The etiology of varicose veins remains elusive. We hypothesized that abnormal cell cycle events in the vein wall may contribute to changes in the structural integrity, thus predisposing to the development of varicosities. The present study was designed to determine whether or not the same molecular apoptotic pathway exists between great saphenous and splenic veins.Methods Thirty-six samples of diseased splenic veins and varicose great saphenous veins were collected. Twenty-five samples of control splenic and great saphenous veins were also collected. The apoptotic cell proteins expression was immunohistochemically stained with antibodies (anti-Bax and anti-Bcl-xl). Apoptosis was evaluated by the terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) assay and immunofluorescence staining. The morphology of apoptotic cells was observed with an electron microscope.Results The apoptotic ratio in walls (intima and media) of diseased splenic vein and varicose great saphenous vein groups were significantly lower than the corresponding regions in the splenic vein and great saphenous vein groups (p<0.01), respectively. A significant difference was not noted in the ratio change of apoptotic cells between the diseased splenic vein and varicose great saphenous vein groups (p>0.05). The high positive expression of Bcl-xl proteins was detected in the diseased splenic vein and varicose great saphenous vein groups, respectively. While the high positive expression of Bax proteins was also observed in the splenic vein and great saphenous vein groups, respectively. Electron microscopic observations confirmed that endothelial and smooth muscle cells in diseased splenic vein, varicose great saphenous vein, splenic vein, and great saphenous vein walls exhibited apoptotic morphologic features, such as fuzzy mitochondrial cristae, medullary changes, and margination of the nuclear chromatin.Conclusions Our results showed the same dysregulation of apoptosis via the intrinsic pathway in diseased splenic veins and varicose great saphenous veins. This observational study suggests that apoptotic down-regulation in the veins wall is a cause of diseased splenic veins and varicose great saphenous veins, but does not exclude the possibility that other mechanisms are involved.

Background: The precise pathogenic mechanism leading to splenic enlargement in patients with portal hypertension is poorly characterized. The present study was designed to determine the expression of nerve fibers and different angiogenic factors in patients with splenomegaly and residual spleens after subtotal splenectomy. Methods: Thirteen cases of splenomegaly due to portal hypertension were examined post-splenomegaly and designated as the splenomegaly group. During the ensuing 8 years we obtained other specimens by puncture biopsy, which were designated as the residual spleen group. We designated 13 patients with traumatic spleen injuries as the control group. Immunohistochemistry and immunofluorescence were used to determine the expression of neuropeptide Y (NPY), neurofilament-200 (NF-200) vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor-2 (VEGFR-2), and CD34 in residual spleens under light microscopy. Results: A significantly higher density of NPY- and NF-positive nerve fibers were observed in the splenomegaly group compared with the residual spleen and control groups (all P<0.05). Moreover, there were no significant differences between the residual spleen and control groups (P>0.05). A significantly higher microvessel density and positive expression ratio of VEGF and VEGFR-2 were observed in the splenomegaly group compared with the residual spleen and control groups (all P<0.05). There were also significant differences between the residual spleen and control groups (P<0.05). Conclusion: The residual spleen can eliminate the high-pressure state after a subtotal splenectomy, and remodeling of splenic nerves and microvessels with other splenic tissues and cells, which contributes to the recovery of splenic function.

INTRODUCTION:Programmed cell death plays a critical role in various physiological processes. In the present study, we investigated its possible pathogenic role in primary varicose veins. We studied histological changes in surgical specimens from thrombophlebitic saphenous veins. In thrombophlebitic saphenous, varicose, and healthy veins, we also determined the number of apoptotic cells, and investigated apoptosis in the role of the pathogenesis of varicose veins. METHODS:Forty-four specimens of thrombophlebitic saphenous veins and simple varicose veins were collected. Thirteen samples of normal great saphenous veins were also collected (control group). Apoptosis of venous walls was determined by terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) and immunofluorescence methods. The corpuscular number per high-power field was counted under light microscopy. RESULTS:A significantly higher apoptotic ratio of the intima and media were observed in control veins as compared with thrombophlebitic saphenous veins and simple varicose veins (p < 0.01). A significant difference was not observed between thrombophlebitic saphenous veins and simple varicose veins (p > 0.05). A significant difference was not seen between the intima and media of the three groups (p > 0.05). CONCLUSION:In the walls of thrombophlebitic saphenous veins and varicose veins, the apoptotic indices were clearly decreased. The results suggest that the process of programmed cell death was inhibited in walls of thrombophlebitic saphenous veins and varicose veins.

Objective Disordered programmed cell death may play a role in the development of superficial venous incompetence. We have determined the number of cells undergoing apoptosis and the alterations in the apoptotic level in the wall of different segments of the great saphenous varicose vein.Methods Twenty-one varicose great saphenous veins (VGSVs) (varicose group) and 12 normal great saphenous veins (GSVs) (control group) were collected, and the apoptosis level in the upper, middle, and lower segments were immunohistochemically stained with antibodies (anti-Bax and anti-Bcl-xl). Apoptosis was evaluated by the TUNEL assay and immunofluorescence staining. The morphology of apoptotic cells was observed with an electron microscope.Results Quantitative analysis showed that the apoptotic ratios in venous walls (intima and media) of the varicose group were significantly lower than the corresponding regions in the control group (all P<0.05). A significantly higher apoptotic rates of the venous walls was observed in control group within the upper compared with the lower segment (P<0.05). Significantly higher positive proteins expression rates of Bcl-xl/Bax were also detected in the VGSVs compared with the GSVs within the three segments, respectively (P<0.01). Electron microscopic observations confirmed that endothelial and smooth muscle cells in varicose and normal vein walls exhibited apoptotic morphologic features, such as fuzzy mitochondrial cristae, medullary changes, and margination of the nuclear chromatin.Conclusion VGSV walls were found to have a significant decrease in apoptotic rate compared with that of GSVs. The rate of apoptosis in the intima and media within the upper segment was increased more than the middle and lower segments in the GSVs. Our findings confirm that programmed cell death is down-regulated in primary varicose veins.

BACKGROUND:Following splenomegaly due to portal hypertension, pathologic characteristics include passive congestion and lymphoplasia. High venous pressure and hemodynamics can result in vascular proliferation and lymphoplasia, and promote splenic microcirculation and functional changes. The aim of this study was to determine the changes in penicillar arterioles (PAs) of red pulp in residual splenic tissue after subtotal splenectomy due to splenomegaly in cirrhotic patients to provide anatomic and physiologic evidence for reserved splenic surgery.METHODS:Thirteen patients with splenomegaly due to portal hypertension, who were treated surgically, comprised the splenomegaly group. After 8 years, we obtained another specimen by puncture biopsy from the residual spleen group. We designated patients with splenic trauma as the control group. The morphology of PAs under light microscopy was facilitated by EVG staining and immunohistochemistry for CD34. Semi-thin sections were HE-stained. The ultrastructure of PA endothelial cells was observed under electron microscopy.RESULTS:In the residual spleen group, diffuse distribution, tenuous elastic intima in the arterial wall, and continuity in PA of red pulp were seen under light microscopy. A significantly lower density and average cross-sectional area of PAs were observed in the residual spleen group compared with the splenomegaly and control groups (P < 0.01). A uniform mitochondrial matrix and a decreased number of ruptured cristae in PA endothelial cells were observed under electron microscopy. While there were some beneficial changes (splenic artery flow volume, portal venous diameter, and portal venous flow volume), the platelet and leucocyte counts were markedly increased in residual spleen.CONCLUSION:Subtotal splenectomy can eliminate the factors which precipitate splenomegaly (portal hypertension), improve the reconstruction of splenic capillaries, correct hypersplenism, and restore normal splenic function.

Background: Portal hypertension (PHT) requires invasive measures to prevent rupture and bleeding of esophagogastric varices; however, the long-term results of subtotal splenectomy plus fixation of the retrosternal omentum majus (SSFROM) have not been reported. Specifically, the advantages and disadvantages of surgery that preserves the spleen and the long-term hematologic effects have not been described.Study design: Our studies relating to SSFROM commenced in February 1999. As of April 2014 we have performed 256 subtotal splenectomies The records of 65 patients with PHT who underwent SSFROM were reviewed retrospectively.Results: Four patients died within 4 years of surgery, with a 4-year survival rate of 94 %; the 11-year survival rate was 60 %. Eleven patients (17 %) had re-bleeding from esophagogastric varices. The white blood cell and platelet counts were higher 6 and 11 years post-operatively compared with pre-operative values (P < 0.01). Portal venous diameter, portal venous flow volume, splenic artery flow volume, as well as splenic length, thickness, and average cross-sectional areas were shown to be significantly constricted or decreased (P < 0.01). The proportion of serum CD3+ T cells, CD4+ T cells, and CD8+ T cells was increased (P < 0.01), while the serum levels of macrophage colony-stimulating factor and granulocyte-macrophage colony-stimulating factor were significantly decreased (P < 0.01). There was no significant change in the serum levels of IgA, IgM, IgG, and Tuftsin (P > 0.05). DSA demonstrated that 15 cases formed collateral circulations between the portal vein and superior vena cava.Conclusion: SSFROM provide long-term hemostasis for esophagogastric variceal bleeding in PHT and corrected hypersplenism. SSFROM is an effective treatment for patients with PHT in whom long-term survival is expected.

目的:观察门静脉高压巨脾大部切除后残脾神经纤维分布与密度变化,评估残脾保留的价值.方法:选取门静脉高压脾肿大行脾大部切除并残脾腹后固定术患者13例,收集患者术后切取的巨脾组织,以及术后8年穿刺获取的残脾组织,另取外伤性脾组织13例为正常对照.采用免疫组化法检测脾神经肽Y (NPY)和神经丝蛋白200 (NF200)阳性神经纤维分布及密度.结果:3组脾组织NPY和NF200阳性神经纤维的分布部位大致相同,但两者在巨脾组织中的密度明显较高.红髓部分的定量分析显示,巨脾组织NPY与NF200阳性神经纤维密度均明显高于残脾组织和正常脾组织(均P＜0.05),而两种阳性神经纤维密度在残脾组织与正常脾组织间差异无统计学意义(均P＞0.05).结论:巨脾大部切除术后残脾神经纤维分布及含量与正常脾大致相同,提示解除高压环境后,残脾神经功能能逐渐恢复正常.

Objective To investigate the changes of penicillar arteriole (PA) of red pulp in residual splenic tissue after subtotal splenectomy due to portal hypertension in order to provide evidence for beneficial outcomes of reserved splenic surgery.Methods We analyzed 13 samples of splenic tissue following surgery for splenomegaly due to portal hypertension as our splenomegaly group.We then took biopsies from the spleens of the same 13 individuals after 8 years to analyze as our residual spleen group.Our control group consist of splenic tissue from 13 traumatic spleen patients.All tissues were then analyzed using EVG staining and immunohistochemistry to observe morphology of penicillar arteriole under light microscope.Results A significantly lower density of PA was observed in residual spleen group compared with splenomegaly group (7.63 ± 0.13 pieces/0.15 mm2 versus 9.53 ± 0.15 pieces/0.15 mm2,P ＜ 0.01),but higher compared with the control group (4.64 ± 0.15 pieces/0.15 mm2,P ＜ 0.01).Significantly lower average cross-sectional areas of penicillar arteriole was observed in residual spleen group compared with both the splenomegaly group (144.32 ± 16.53 μm2 versus (323.52 ± 99.31) μm2,P ＜ 0.01) and control group (185.0123.65 μm2,P ＜ 0.01).Conclusion Subtotal splenectomy attenuates initiation factors of splenomegaly due to portal hypertension,improves reconstruction of splenic capillaries,and promotes retension of immune function of the residual spleen.

BACKGROUND:In recent years, the spleen has become to be recognized as the "control center" of the immune-metabolic-endocrine network. However, It is controversial that splenomegaly due to portal hypertension is treated by subtotal splenectomy. The aim of this study was to evaluate the distribution of fibrous tissue, morphology of cells as well as splenic size, hemodynamics, hematological and immunological indexes in the residual spleen after subtotal splenectomy. This information may help finding the basis for the operation of subtotal splenectomy.METHODS:Ten cases of splenomegaly due to portal hypertension were investigated. Two groups were created: Splenomegaly and Residual spleen. Control group was 10 cases of trauma-induced splenic rupture. Samples were sliced, and morphological changes were observed under light microscopy and electron microscopy. Indexes of splenic size, hemodynamics, hematology and immunology of the spleen were measured.RESULTS:Under light microscopy, the number of collagen fibers and elastic fibers was increased, and the number of reticular fibers was decreased in the residual spleen and splenomegaly groups. Under electron microscopy, the ultrastructure of endothelial cells, lymphocytes, macrophages, and reticular cells in the residual spleen group were noticeably improved more than in the splenomegaly group. Flow volume in the residual spleen and portal vein decreased obviously, with number of platelet rising to normal, and there was no significant difference in the indexes of immunology.CONCLUSION:After subtotal splenectomy, the residual spleen will not experience a high-pressure environment, and the fibrosis of splenic tissue and remodelling of corpuscular morphology will cease.

The spleen is thought to be central in regulating the immune system, a metabolic asset involved in endocrine function. Overwhelming postsplenectomy infection leads to a mortality rate of up to 50%. However, there is still controversy on performing subtotal splenectomy as treatment of splenomegaly due to portal hypertension in cirrhotic patients. In the present study, immunocytes and the indexes of splenic size, hemodynamics, hematology and immunology in the residual spleen were analyzed to support subtotal splenectomy due to splenomegaly.

Objective: To observe the apoptotic level alterations in the wall of different segments of great saphenous varicose vein. Methods: Twenty great saphenous varicose veins(observation group) and 12 normal great saphenous veins(control group) were collected,and the apoptosis in their upper,middle and lower segments were determined by TUNEL assay and immunofluorescence staining,respectively. Results: In observation group,only scattered apoptotic cells were found in the venous walls(tunica intima and tunica media),which were almost uniformly distributed at the three segments,while there were considerably more apoptotic cells in the venous walls(tunica intima and tunica media) in control group,and the apoptosis was more evident in the upper segments than those of the lower segments.The quantitative analysis showed that the apoptotic rates in either the tunica intima or tunica media in observation group were significantly lower than those in their corresponding regions in control group(all P0.05);statistical significances in apoptotic rate were found between some of the segments after intra-group comparison for each group(P0.01 or P0.05),but generally,the apoptotic rate was higher in the upper segment than that in the lower segment and higher in the tunica intima than that in the tunica media. Conclusion: In the wall of great saphenous varicose vein,the apoptotic level is decreased,and there was no apoptotic rate difference between segments in the normal great saphenous vein.

Objective To investigate the changes of collagen and elastic fibers in hypertensive splenic vein and varicose great saphenous vein wall media, in order to compare the degree of collagen and elastic fiber remodeling in the two kinds of veins. Methods Four groups were divided, including Splenic veins of patients with portal hypertension (n=20) and control group (n=20), varicose great saphenous veins (n=20) and control group (n=20) were collected and stained with elastica van Gieson (EVG), the distribution of collagen and elastic fibers in vein walls was observed, and the relative content of collagen (RCC). The relative content of elastic fibers (RCE) were measured with light microscope. Results RCC and RCE were (50.75±14.79)% and (11.36±4.03)% in splenic vein group, (31.22±5.71)% and (14.60±2.95)% in control group, respectively. For RCC and RCE, there were significant differences between the splenic vein group and control group (RCC: P < 0.01 , RCE: P < 0.05). RCC and RCE were (29.71±6.63)% and (7.50±2.49)% in varicose group, (23.52 ± 6.20)% and (9.71 ± 3.89)% in control group, respectively. The difference of RCC was statistically significant (P < 0.05). For change ratio of RCC, there were statistically significant differences between the splenic vein groups and varicose group (P < 0.01). Conclusion Collagen fiber content increased significantly and elastic fibers reduced in splenic vein and great saphenous vein wall media under the state of venous hypertension. And the ratio of increase in splenic veins is significantly more than that in great saphenous veins.

The purpose of this study is to describe the infiltration of mast cells as well as T and B cells in the walls of thrombotic varicose great saphenous veins. Sections were obtained from venous segments of patients with varicose veins and stained with toluidine blue for mast cells, while immunohistochemistry for T cells (using CD45RO antibody) and B cells (CD20) was analyzed using light microscopy after staining. The number of mast cells, T, and B cells observed in thrombotic varicose veins was 1.925 ± 1.203, 72.038 ± 34.707, and 19.519 ± 9.899, respectively. In varicose veins, the corresponding values were 0.265 ± 0.099, 0.600 ± 0.432, and 0.488 ± 0.400. Significantly higher number of mast cells, T cells, and B cells were observed in thrombotic varicose veins compared with control veins. A significant difference was not observed between the varicose group and control group. Thrombi in varicose veins can induce infiltration of mast cells, T cells, and B cells, which may be involved in the remodeling of venous walls.

有研究表明,下肢曲张静脉管壁的改变先于瓣膜功能不全之前[1].细胞凋亡参与曲张静脉管壁重塑过程,两者之间关系密切[2-3].我们通过原位细胞凋亡(TUNEL)法和免疫荧光法对3组下肢静脉管壁细胞凋亡进行定量分析. 一、资料与方法 1.一般资料:选取解放军第89医院普外科2009年10月至2011年10月行高位结扎剥脱加旋切治疗的大隐静脉曲张患者44例,并分为浅表血栓性静脉炎(血栓组)22例和单纯静脉曲张(曲张组)22例.对照组为创伤行截肢术而大隐静脉无损伤者13例.血栓组为慢性血栓性静脉炎(血栓形成时间＞30 d).曲张组按CEAP分类选取C2 ～ C3.

Objective To investigate the distribution and the content of macrophages (mΦ) in the residual splenic tissue after subtotal splenectomy.Methods Thirteen patients with splenomegaly due to portal hypertension were studied.The specimens were obtained by operation or puncture biopsy.There were 2 groups:the splenomegaly and the residual spleen groups.The control group consisted of 13 patients with trauma-induced splenic rupture.The samples were sliced,and the distribution and counts of mΦ were observed under light microscopy using immunohematological staining.Results The mΦ of the residual spleen were equally distributed in the splenic cord and they adhered to the wall of the splenic sinus in the red pulp.They had a high density and were deeply stained.In the white pulp,the mΦ were scattered in the splenic corpuscle and were significantly concentrated in the marginal zone (MZ),where larger mΦ were found.There was a significantly higher number of mΦ in the red pulp of the residual spleen group when compared with the splenomegaly and the control groups.There was a significantly higher number of mΦ in the red pulp of the control group (7.46±0.86) when com pared with the splenomegaly group (4.20±0.88) or with the residual spleen group (31.09±9.73).A significant difference was not observed among the three groups in the white pulp (splenic corpuscle and MZ).Conclusions After subtotal splenectomy for splenomegaly due to portal hypertension,the number of unit area of mΦ in the red pulp in the residual spleen increased significantly,while the number of mΦ in the white pulp did not change much in the three groups of patients.The results suggested that residual spleens still have good phagotrophic functions.

Objective To investigate the changes in collagen,elastic and reticular fibers in the splenic remnant after subtotal splenectomy for splenomegaly,and to study for the basis for spleenpreservation technique for this condition.Methods We studied 13 patients with splenomegaly due to portal hypertension.The specimens were obtained by operation and puncturing biopsy,and they were divided into the splenomegaly group and the splenic remnant group.The control group consisted of 13 patients with traumatic splenic rupture.Using Masson staining,EVG staining,and Silver staining,the samples were analyzed using light microscopy after staining.Results The relative contents of collagen fibers (RCC),elastic fibers (RCE),and reticular fibers (RCR) in the splenic remnant,the splenomegaly,and the normal spleen were (7.81 ±0.83)％,(7.72± 3.42)％,(4.67 ± 1.09)％;(7.76±0.81)％,(7.21±3.18)％,(4.79±1.20) ％ ; (3.21±0.51)％,(1.67±3.22)％,(10.92±0.86)％,respectively.There was no difference between the splenomegaly and the splenic remnant for the three kinds of fiber content (P＞0.05).Significantly higher numbers of collagen and elastic fibers were observed in the control group when compared with the splenic remnant and the splenomegaly groups (P＜0.05).Significantly lower numbers'of reticular fibers were observed in the splenic remnant and the splenomegaly groups when compared with the control group (P＜ 0.05).Conclusions As the remnant spleens were fixed in retropertoneum after subtotal splenectomy,there could not be excessive fibrosis in the splenic tissues.

【Objective】 To investigate the effects of the apoptosis of the walls of the great saphenous and splenic vein under high hemodynamic pressure. 【Methods】 36 samples of varicose great saphenous veins and diseased splenic veins were collected. 25 samples of normal great saphenous veins and splenic veins were al- so collected(control group). Apoptosis of venous walls were determined by the TUNEL and immunofluores- cence method. The number of cells per high-power field was counted under light microscopy. 【Results】 Sig- nificantly higher apoptotic ratio of the intima and media were observed in control veins as compared with varicose veins and diseased splenic veins(P <0.01). A significant difference was not observed between the in- tima and media(P >0.05). 【Conclusion】 In the walls of the great saphenous vein and splenic vein under high venous pressure, the apoptotic indexes were decreased obviously. The results suggest that can affect the apoptotic regulation of the wall by high hemodynamics.

Objective To investigate the infiltrative changes of T and B lymphocytes in residual splenic tissues after subtotal splenectomy for splenomegaly.Methods Thirteen patients with splenomegaly due to portal hypertension were studied.The specimens were obtained by operative and percutaneous puncture biopsy.There were 2 groups of specimens from this group of patients:the splenomegaly and residual spleen groups.The control group consisted of 13 patients with traumatic rupture splenic tissues.The samples were sliced,and the distribution and counts of T and B lymphocytes were observed under light microscopy using immunohematological staining.Results In the residual spleen group,the numbers of T and B cell were (59.769±9.429) per 10 5 pixel area,(10.822±1.938) per 10-5 pixel area,while in the splenomegaly group they were (34.715 ± 3.497) per 10-5 pixel area,(2.369±0.664) per 10-5 pixel area,and in the control group they were (48.229±14.869) per 10-5pixel area,(6.844±0.807) per 10-5 pixel area,respectively.A significantly higher number of T and B cells were observed in the residual spleen group when compared with the splenomegaly group and the control group (P＜0.05).Significantly lower numbers of T and B cells were observed in the splenomegaly group when compared with the control group (P＜0.05).Conclusions T and B lymphocytes increased significantly per unit area in the residual spleen after subtotal splenectomy.The results suggested that the immune function of the residual spleen can be improved.

Objective To investigate the changes of pathomorphological features and lumen diameter of varicose great saphenous veins wall with thrombus. Methods Eighteen cases of varicose great saphenous veins with thrombus(thrombus group), 18 cases of simple varicose veins(varicose group), and 12 cases of normal great saphenous veins (control group) were collected. Using hemotoxylin and eosin staining and Masson staining, the wall structure was observed, and the vein diameter and the thickness of the inner, middle and outer membranes were measured. Results In thrombus group, a large number of inflammatory cells were observed in full-thickness of venous wall, endothelial cells were lost on adhesion of thrombus, and partal thrombus organization was caused by a lot of collagen fibers extending from rupture of intima to introthrombus. In varicose group,inflammatory cells were observed occasionally in full-thickness of venous wall, intimal thickened, smooth muscle cells in mid layer were proliferated irregularly and disordered, and collagen fibers proliferated. In control group, intima of venous wall was thin, the medial smooth muscle bundles was regular on arrangement. The lumen diameter, the thickness of the inner,middle and outer membranes thrombus group were significantly higher than that of varicose group and control group (P<0.05), while the lumen diameter and intima thickness of varicose group were significantly higher than that of control group(P<0.05). Conclusion Lumen increasing and wall thickening are important features of path morphological changes.