Purpose To study changes of cell cycle of vascular endothelial cell in non-proliferative diabetic retinopathy. Methods Alloxan induced Wistar-rats were employed and immunohistochemistry,Western blotting methods were used. Results The vascular endothelial cells of retinas of 8～20 weeks diabetic rats were observe to be cyclinD1,cyclinD3,cyclinB1,p21 and p27 positive stained with light and electronmicroscopies.CyclinE immuno-stained vascular endothelial cells was observed occasionally.Meanwhile,the evidences of morphologic changes of the vascular en dothelial cells were proved:less plasma,thinner cell,more bubble organelles than those of controls.But,the ultra-structures of pericytes and other type of retinal cells did not change and they also immunostain negative.Komas blue and Western blotting methods also proved that the vascular endothelial cells of retina of 20th week diabetic rats expressed cyclinD1,cyclinB1,p21 and p27 protein. Conclusion Glucose induced retinal vascular endothelial cells of 8～20th weeks diabetic rats enter cell cycle and were arrested at G1/S restriction point.This study also suggested that retinal vascular endothelial cells may possess the ability to resist glucose damage and mechanism of selfstability during very early stage of diabetes. (Chin J Ocul Fundus Dis,2000,16:173-176)
目的:探讨胚胎发育不良性神经上皮肿瘤(DNT)的临床、影像及病理学特征、诊断及鉴别诊断.方法:回顾性分析8例胚胎发育不良性神经上皮肿瘤患者的临床和影像学资料,进行光镜和免疫组织化学染色观察,并获得6例的随访资料.结果:胚胎发育不良性神经上皮肿瘤男性7例,女性1例,年龄为5～19岁,平均年龄13岁,5例以癫痫小发作为主要临床表现,病变均位于幕上,以皮层为主,影像学检查均无明显的占位效应及瘤周水肿。肿瘤细胞主要由少突胶质样细胞(OLC)、神经元和星形细胞组成,4例伴有皮质发育不良。免疫组织化学结果为神经元及部分少突胶质样细胞呈嗜铬素A、突触素及S-100阳性表达;少突胶质样细胞呈胶质纤维酸性蛋白(GFAP)阴性表达,而星形细胞呈GFAP阳性表达;Ki-67抗原标记指数lt;1%。结论: 胚胎发育不良性神经上皮肿瘤为WHOⅠ级良性肿瘤,可结合临床、影像及病理学表现明确诊断,预后良好,无需放疗和化疗。
Objective To investigate the pathological changes in the neuromuscular junction during ischemiareperfusion(IR) in the skeletal muscle. Methods Forty-eight healthy adult Wistar rats (24 male, 24 female) were equally randomised into the following 6 groups: Group A (control group): no ischemiareperfusion; Group B: ischemia by clamping the blood vessels of the right hindlimb for 3 hours; Group C: ischemia by clamping for 4.5 hours;Group D: ischemia by the clamping for 4.5 hours followed by reperfusion for 1.5hours; Group E: ischemia for 4.5 hours followed by reperfusion for 24 hours; and Group F: ischemia for 4.5 hours followed by reperfusion for 2 weeks. Then, the medial head of the gastrocnemius muscle flap model was applied to the right hindlimb of each rat. The medial head of the gastrocnemius muscle was isolated completely,leaving only the major vascular pedicle, nerve and tendons intact.The proximal and distal ends (tendons) were ligated while the vessel pedicle was clamped. And then, Parameters of the muscle (performance,contraction index,colour,edema,bleeding) were observed. The muscle harvested was stained with gold chloride(AuCl3) and the enzymhistochemistry assay (succinate dehydrogenase combined with acetylcholine esterase) was performed. Morphology and configuration of the neuromuscular junction were observed during the ischemiareperfusion injury by means of the AuCl-3 staining. The result of the enzymhistochemical reactions was quantitatively analyzed with the computer imageanalysis system. And then, additional 5 rats were prepared for 3 different models identical with those in Groups A, C and E separately. The specimens were harvested from each rat and were stained with HE and AuCl-3, and they were examined under the light microscope. Results During the period of ischemia, the skeletal muscle of Group B showed the colour of purple and edema.The colour and edema became worse in Group ,while dysfunction of elasticity and contraction appeared obviously with plenty of dark red hemorrhagic effusion at the same time.After reperfusion,the color and edema of muscle in Group D became improved while the elasticity and function of contraction was not improved. Hemorrhagic effusion of Group D turned clearer and less than Group C.Group E was similar to Group D in these aspects of muscle except for much less hemorrhagic effusion. Skeletal muscle in Group F showed colour of red alternating with white, adhesion,contracture of muscle, exposure of necrotic yellow tissue and almost lost all its functions. The AuCl3 staining showed that during IR, necrosis of the myocytes was followed by degeneration of their neuromuscular junctions, and finally the nerve fibers attached to these neuromuscular junctions were disrupted like the withering of leaves. The enzymhistochemistry assay showed thatthere was no significant difference in the level of acetylcholine esterase between the ischemic group (Groups B and C) and the control group (Group A) (Pgt;0.05). However, the level of acetylcholine esterase in all the reperfused groups (Groups D, E and F) decreased significantly when compared with the control group（Group A）and the ischemic groups (Groups B and C) (Plt;0.01). Conclusion The distribution of the nerve fibers and the neuromuscular junctions in the mass of the muscles is almost like the shape of a tree. The neuromuscular junction seems to be more tolerant for ischemia than the myocyte. Survival ofthe neuromuscular junction depends on its myocytes alive. Therefore, an ischemiareperfusion injury will not be controlled unless an extensive debridement of the necrotic muscle is performed.
ObjectiveTo investigate the expression of caspase-3 and Toll-like receptor 4 (TLR4) in the incised rat skin healing process and its relationship with the wound time and to provide an experimental evidence for the prediction of injury time. MethodsAfter the rat incised wound model was established, hematoxylin-eosin dyeing technology and immunohistochemical staining technique were used to observe the expression of caspase-3 and TLR4. Then Image Pro Plus Image analysis software and SPSS statistical analysis software were used to deal with the experimental results. ResultsCaspase-3- and TLR4-positive cells were detected in epidermis, hair follicle and sebaceous gland cells in the control skin. The expression of caspase-3 and TLR4 of the ante mortem groups were significantly different compared with the control group except the 0 h group (P<0.05). Caspase-3- and TLR4-positive cells were detected in neutrophils around the hair follicle half an hour later. Caspase-3- and TLR4-positive cell rate increased with the infiltration of inflammatory cells. Caspase-3- and TLR4-positive cell rate reached the maximum on the 3 rd day, and then it began to decrease, and they were mainly expressed in fibroblasts and mononuclear macrophages. Caspase-3- and TLR4-positive cells were mainly expressed in fibroblasts on the 10th day. There was no significant differences between the postmortem injury groups and the normal control groups (P>0.05). ConclusionCaspase-3- and TLR4-positive cell rate is time dependent and stable in 25℃ temperature environment which makes it possible to determine the time of injury.
The effects of preoperative selective arterial perfusion chemotherapy (PSAPC) in the treatment of 20 cases of histologically confirmed gastric cancers is reported in which 12 patients were operated on, and a comparison with that of general chemotherapy in 10 cases of gastric cancers (as controls) was made. In the PSAPC group, besides the improvement of clinical symptoms and singns, the cancer cells of lesions in situ and metastatic lymph nodes have different degrees of degeneration and necrosis. This result show significant differences in two group comparison (Plt;O.01). We conclude that the PSAPC has good short-term effect and little side-effect. It can improve the resection rate and radical resection rate, and prevent the iatrogenic metastasis and implantation during operation, and decrease the postoperative recurrence of the patients with gastric cancer.
Objective To observe the configuration and viability of full thickness human fetal retina after short-, mid- and long-term preservation. Methods Twenty-two full thickness human fetal retinae of gestational age of 12-24 weeks were coated by glutin and cut into 88 pieces, and then preserved in Ames' solution, DX solution, -80℃ refrigerator or under cryopreservation condition. The cell viability of retinal neuroepithelial layer was determined by trypan blue staining, retinal configuration was determined by light microscope and electromicroscope. Results The viability of neuroepithelial layer was (94.79plusmn;2.85) % in fresh fetal retina, gt;80% in Ames' solution within 4 hours, and gt;77% in DX solution within 2 days. There was no significant difference between those solution-preservations and the fresh fetal. In -80℃ refrigerator, the viability was (65.83plusmn;5.06)% after 7 days, and then dropped to (57.54plusmn;16.18)% at the end of the first month. Under the cryopreservation condition, the viability was (69.46plusmn;9.31)% at the end of first month. Light and transmission electron microscopy had not deteced any abnormals in the full thickness human fetal retina preserved in Ames' solution within 2 hours, but showed clear retinal layers with bigger intercellular space after preserved in DX solution for 2 days, in -80℃ refrigerator for 7 days and under cryopreservation condition for 1 month. Conclusion Ames' solution and DX solution can preserve good viability and configuration of full thickness human fetal retina in a certain time period.
Objective To observe the classification,clinical and pathological features of ciliary body tumors. Methods The clinical and pathological data of 11 cases of primary ciliary body tumors were analysed retrospectively. Results By pathological examination,the tumors of these cases were devided into malignant melanoma (2 cases), benign melanocytoma (3 cases),leiomyoma (2 cases), and angio-leiomyoma, neurilemoma, non-special granuloma and medulloepithelioma (1 case respectively). Both of the benign and malignant tumors of ciliary body tended to grow and enlarge progressively. The cardinal clinical manifestations of this series of 11 cases were as follows: elevation of intraocular pressure in 7, local scleral vascular dilatation in 5, secondary exudative retinal detachment in 5,and the signs of anterior uveitis in the early stage of tumor growth in 4. Conclusion The histopathological types of cilliary tumors are manifold,and the tumors are prone to enlarge progressively in developement either in benign or malignant ones, so that the rates of clinical misdiagnosis are relatively high. (Chin J Ocul Fundus Dis, 2002, 18: 273-275)
To investigate the relationship between clinicopathological features and lymph node metastasis in the primary gastric cancer and affer the basis for deciding appropriate extent of lymph node dissection, a total of 192 patients who underwent curative gastrectomy and lymph node dissection for gastric cancer were analyzed retrospectively. Result: The total rate of lymph node metastasis was 60.4%, with 28.9% of the resected lymph nodes involved. The lymph node metastasis of C, M, A region and the whole stomach were 64.6%, 57.7%, 59.1% and 90.9% respectively. The rates of the lymph node metastasis increased successively in carcinoma of early, middle and late stages (Ｐ＜0.05), the rate of the infiltrative tumor (Borr Ⅲ，Ⅳ) being 76.5% which was significantly higher than that of the circumscribed tumor (Borr Ⅰ，Ⅱ) (43.2%)。 Relating with the tumor size ＜4cm in diameter showed lesser rate, while 4-8 cm and ＞8cm in diameter showed increasingly higher metastaticrate (Ｐ＜0.01). As a result, we should decide the appropriate extent of lymph node dissection during the operation on the basis of clinicopathological stages, type of Borrmann’s, site and maximum diameter of gastric cancer along with the state of lymph node metastasis in carcinoma of different region of the stomach.