ABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of LPS on the expression of CD14 and the activation of Kupffer cells (KCs).</p><p><b>METHODS</b>Rat KCs were isolated and cultured with LPS. Immunohistochemistry and RT-PCR methods were employed to determine the changes in the CD14 expression and the concentration of TNFalpha, IL-6 and NO in the supernatant of the cultured KCs with LPS.</p><p><b>RESULTS</b>(1) The expression of CD14mRNA and the synthesis of CD14 protein in the KCs increased evidently when stimulated by various concentrations of LPS, and the CD14mRNA expression was correlated in dose-dependent manner with LPS levels. (2) The expression of CD14mRNA and the synthesis of CD14 protein in KCs induced by LPS (10 micro g/ml) increased significantly and peaked at 3 approximately 6 hours. (3) The expression of CD14mRNA and the synthesis of CD14 protein in freshly cultured KCs were obviously up-regulated by the active mediators produced by KCs after being stimulated by LPS. (4) The release of TNFalpha, IL-6 and NO from cultured KCs was evidently down-regulated by the addition of anti-CD14McAb in the presence of serum or by the addition of LPS in the absence of serum, but up-regulated by the concomitant addition of LPS and LBP.</p><p><b>CONCLUSION</b>(1) The CD14mRNA expression and the protein synthesis in cultured KCs were closely related to LPS and the active mediators produced from the KCs.The increased CD14 expression was possibly caused by LPS, and the further increase of the expression might be closely correlated to the cytokines released from the KCs. (2) The KC activation by low concentration of LPS was CD14 dependent.</p>
Subject(s)
Animals , Rats , Cells, Cultured , Dose-Response Relationship, Drug , Gene Expression Regulation , Interleukin-6 , Bodily Secretions , Kupffer Cells , Cell Biology , Metabolism , Lipopolysaccharide Receptors , Genetics , Metabolism , Lipopolysaccharides , Pharmacology , Nitric Oxide , Bodily Secretions , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Tumor Necrosis Factor-alpha , Bodily SecretionsABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of postburn fluid resuscitation on the pathohistological and ultrastructural changes of multiple organs with dysfunction in severely burned dogs.</p><p><b>METHODS</b>Forty - four mongrel dogs were randomly divided into four groups: (1) immediate infusion (II, n = 8), (2) delayed infusion (DI, n = 15), (3) no infusion (NI, n = 14), (4) normal control (NC, n = 7). The dogs were inflicted with 50% TBSA III degree flame burn produced by napalm in concentration of 30g/L burning for 30 seconds on the back. Small pieces of tissue samples of heart, lungs, liver, kidneys and gastrointestinal tract were taken from injured dogs at 72 postburn hours (PBHs) or moribund stage for the examination with light microscope (LM) and transmission electron microscope (TEM).</p><p><b>RESULTS</b>Different degrees of blood circulation disturbance and degenerative changes were found in all above internal organs. These changes were more evident in DI than in II and NI groups.</p><p><b>CONCLUSION</b>Delayed postburn fluid resuscitation could induce multiple organ dysfunction in early postburn stage.</p>
Subject(s)
Animals , Dogs , Burns , Therapeutics , Digestive System , Pathology , Fluid Therapy , Kidney , Pathology , Liver , Pathology , Lung , Pathology , Multiple Organ Failure , Pathology , Myocardium , Pathology , Time FactorsABSTRACT
Objective To observe the effects of advanced glycated bovine serum albumin(AGE-BSA) on the tubule like structure(TLS) formation of endothelial cells.Methods There were three groups:AGE-BSA(25,50 and 100 ?g/ml) treated groups,BSA(25,50 and 100 ?g/ml) treated groups as BSA control groups,endothelial complete medium without AGE-BSA or BSA as blank control groups.After incubation of human umbilical vein endothelial cells(HUVECs) for 24,48 and 72 h,TLS formation assay and ELISA were used to detect the length of TLS formation on the surface of collagen Ⅰ and the vascular endothelial growth factor(VEGF) autocrine of HUVECs in presence of AGE-BSA.Results In 24 to 72 h,AGE-BSA(25,50,100 ?g/ml) increased the average length of TLS formation on the surface of collagen Ⅰ and the concentration of VEGF autocrine in supernatant of HUVECs(P
ABSTRACT
Objective To investigate the effects of the new drug Nordy on malignant glioma cells induced tubule-like structure formation of endothelial cells in a three dimensional culture system with collagen coating.Methods The endothelial cells were divided into three groups.Group A were cultured on the DMEM media coated with collagen;group B were cultured in 2:1(V/V) DMEM to the supernatant of malignant glioma cell culture;group C,based on the group B,were added with Nordy of different concentrations.The cell cycle and apoptosis of ECV304 cells were determined and the process of tubular formation were observed.Moreover,the parameters such as the forming curves,outer and inner diameter,wall thickness and length of the tubules were measured and compared between groups B and C.Results The cell cycle of endothelial cells were restrained,and some cells were induced to apoptosis.The tubule-like structure formation was inhibited,and the tubules in group B appeared earlier,larger in diameter and longer in length than group C.Conclusion The tubular formation induced by malignant glioma cells could be restrained by the new drug Nordy.
ABSTRACT
Objective To observe the effects and significance of Nordy on the expression of vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS) in the retinas of diabetic rats. Methods Diabetic rats by streptozotocin were randomly divided into diabetes group (5 rats?2) and Nordy treatment group (5 rats?2). Another 10 normal rats were recruited as normal control group. The Nordy treatment group was injected 0.5% Nordy (27 mg/kg) while diabetes groups and normal control group were injected saline solution into peritoneal cavity once every other day. One month or 3 months later, 5 rats in each group were killed and the expression of VEGF and iNOS in the retinas were detected by immunohisochemistry. The average positive areas were measured and analyzed by computer aided video system. Results The expression of VEGF and iNOS in control group were extremely low. In 1 month, the expression of VEGF and iNOS in diabetic group increased and the average positive areas of VEGF and iNOS were significantly more than that of Nordy treatment group (P
ABSTRACT
Objective To investigate roles of cyclin-dependent kinase 4 (CDK4) in nordihydroguaiaretic acid (NDGA)-induced inhibitory effect on proliferation of human malignant glioma cells. Methods The techniques of cell culture, cell counts, flow cytometry, immunoprecipitation, immunohistochemistry and image analysis were employed in this study. Results ①A concentration-dependent inhibition of proliferation was demonstrated in the SHG-44 cells incubated for 24 hours in the presence of NDGA, and cell proliferation was blocked in the G1→S phase. ②The activity of CDK4 was decreased apparently in the SHG-44 cells treated for 24 hours with 10 to 200 μmol/L NDGA in a concentration-dependent way. ③The expression of CDK4 gene was downregulated in the cells after NDGA treatment. Conclusion CDK4 plays an important role in NDGA-induced inhibition of glioma cell proliferation.
ABSTRACT
Objective To investigate the changes and their significance of bcl-2 and c-myc in nordihydroguaiaretic acid (NDGA)-induced apoptosis of human malignant glioma cell line SHG-44. Methods The apoptosis of SHG-44 cells was observed with light and electron microscopy and TUNEL method. The expression of bcl-2 and c-myc gene was measured with immunohistochemistry, in situ hybridization and image analysis. Results ① The SHG-44 cell apoptosis was induced by NDGA at a concentration lower than 200 μmol/L in a time-dependent manner. ② The expressions of bcl-2 and c-myc gene in SHG-44 cells were decreased after the treatment of 100 μmol/L NDGA with the elapse of time, indicating a close association with cell apoptosis. ③ The expressions of bcl-2 and c-myc mRNA in SHG-44 cells were decreased after the treatment with 100μmol/L NDGA, which was apparently consistent with the immunohistochemical results. Conclusion The NDGA-induced apoptosis in human malignant glioma cells might be related with the down-regulated expressions of bcl-2 and c-myc gene. The exact mechanism needs further research.
ABSTRACT
Objective To explore the effect of nordihydroguaiaretic acid (NDGA) on the expressions of vascular endothelial growth factor (VEGF) and its receptor, kinase-inserted domain containing receptor(KDR) and the possible mechanism. Methods The expression of VEGF in human malignant glioma cell line SHG-44 and that of KDR in human umbilical vein endothelial cell (HUVEC) line ECV-304 were observed 1～3 d after NDGA treatment with immunohistochemistry, in situ hybridization and image analysis. Results The expression of VEGF was declined at protein or mRNA levels in SHG-44 cells after treated with 100 μmol/L NDGA for 1 to 3 d. The expression of KDR in endothelial cells with 100 μmol/L NDGA treatment for 1 to 3 d was decreased too, in a more obvious way compared with the decline of VEGF expression in SHG-44 cells. Conclusion The results suggest that NDGA inhibits the expression of VEGF in glioma cells as well as that of VEGF receptor KDR in endothelial cells, which may be the important molecular mechanism of anti-angiogenesis of NDGA.
ABSTRACT
Objective To investigate the effects of TNP-470 on the growth of a human malignant glioma cell line SHG-44 in vivo and in vitro. Methods The colorimetric MTT assay, soft agar culture, flow cytometry，light and electron microscopy were used to determine the proliferation, the cloning efficiency, cell cycle and the morphological changes of SHG-44 cells as well as the growth of its xenografted tumor. Results TNP-470 (20～2 000 ng/ml) significantly inhibited the proliferation of SHG-44 cells in vitro (the 50% inhibitory concentration was 200 ng/ml). Cloning efficiency reduced obviously. The number of cells in G0/G1 phase increased, while that in S, G2/M phases decreased significantly. Weight and volume of xenografted tumors treated with TNP-470 (30 mg/kg, injected subcutaneously every other day) reduced notably. Furthermore, there were necrotic area and apoptosis in the tumor. No severe side effect of TNP-470 was found in this study. Conclusion The inhibitory effect of TNP-470 on the growth of SHG-44 cells correlates with its functions of regulating cell cycle and inducing apoptosis, which suggests that the angiogenesis inhibitor TNP-470 has strong inhibitory effect on human malignant gliomas.
ABSTRACT
Objective To investigate morphological changes of endothelial cells after nordihydroguaiaretic acid (NDGA) treatment in vitro. Methods The morphological changes of human umbilical vein endothelial cells (HUVEC) cell line ECV-304 and the cell apoptosis rate in sub-G0 phase were observed with invert, light and electron microscope and flow cytometry after NDGA treatment at different concentrations or with PBS (0.01 mol/L) as control. Results ①After the treatment of NDGA at 50～200 μmol/L for 1～3 d or up to 8 d at 100 μmol/L, ECV-304 cells tended to elongate into a shuttle-like sparse appearance and those in mitosis were decreased, indicating the suppression of cell proliferation. All these alteration was in a time-and dose-dependent manner. ②NDGA-treated ECV-304 cells displayed morphological features of apoptosis, especially at the 48th h after the treatment. With flow cytometry, the cells in sub-G0 phase were significantly increased, and reached its peak at hour 12 (20.42%) after NDGA treatment. In addition, the degeneration and necrosis of ECV-304 cells were related to the concentrations of NDGA. Conclusion NDGA can inhibit the proliferation and growth of endothhelial cells, and induce apopotosis, which might also inhibit angiogenesis.
ABSTRACT
Objective To explore the relationship between morphologic evolution and proliferative activity during hepatocarcinogenesis in rats. Methods Imaging analysis technique was used to detect the morphologic parameters of cells in hepatic lesions in both Solt-Farber model and diethylnitrosamine (DEN)-induced liver cancer model. Immunohistochemistry was employed to detect proliferating cell nuclear antigen (PCNA) and bromodeoxyuridine (BrdU). Results The oval cells were identified as irregular small proliferating cells in size of one-eighth of and with a nucleus/cytoplasm ratio of 6 times of the normal hepatocyte by image analysis. The morphometric parameters of basophil hepatocyte in precancerous foci and nodule were similar to those of the liver cancer cell. PCNA and BrdU positive cells were mainly localized within the proliferative foci and hepatocellular carcinoma (HCC) tissues. There was a better consistency between the development of hepatic lesions and cellular proliferative activity. Conclusion The morphologic evolution is closely related to proliferative activity during hepatocarcinogenesis in rats.
ABSTRACT
Objective To study the expressions of oncogenes c-Ha-ras, c-ki-ras, pan-ras and c-myc and point mutation of c-Ha-ras1 during hepatocarcinogenesis in rats. Methods Immunohistochemistry, in situ hybridization and microdissection of tissue (MDT)-PCR-SSCP were used to detect the oncogene expressions and point mutation of c-Ha-ras1 in both Solt-Farber model and DEN-induced liver cancer model. Results The overexpression of c-Ha-ras was closely associated with the formation and proliferation of the precancerous basophilic hepatocyte foci, while that of c-myc with the growth of the oval cell foci. The abnormalities of IGF-Ⅱ played an important role in the evolution of precancerous foci/nodules towards hepatocellular carcinoma (HCC). The overexpression of fms was only associated with HCC of some rats. Conclusion Hepatocarcinogenesis in rats was related with the overexpression of c-Ha-ras, c-myc, IGF-Ⅱand fms and the point mutation of c-Ha-ras1, and overexpression of these oncogenes was associated with morphological evolution.
ABSTRACT
AIM: To study the changes in myocardial blood flow (MBF), capillarization and cardiac function in the rat during acclimation to hypoxia. METHODS: Myocardial capillary density (CD) and capillary/myocyte ratio (C/M) was assayed by alkaline phosphatase histochemistry. Biomicrosphere method was used to determine MBF in the rat after 5, 15 or 30 days hypobaric hypoxic exposure (5 000 m). RESULTS: In the course of hypoxia, MBF and cardiac function increased in the right ventricle. However, in the left ventricle, acute hypoxia caused an increase in MBF and a decrease in cardiac function. Both returned to the control level on continued hypoxic exposure. Neovascularization occurred after 15 day or 30 day of hypoxic exposure in both ventricles, judged from the significant increment of C/M ratio albeit the CD remained unchanged in the right ventricle. CONCLUSION: Our findings indicate that adaptive changes in rat heart during acclimation to hypoxia include: ① persistent increase in MBF, hypertrophy associated with increase in capillarity and enhanced cardiac function of the right ventricle; ② increase in MBF and depression of cardiac function at first, then followed by recovery of MBF and increase in capillarity accompanied with recovery of left ventricular function.
ABSTRACT
AIM: To compare the effects of hypoxia on expression of vascular endothelial growth factor (VEGF), iNOS and eNOS mRNA in cultured umbilical vein endothelial cells (UVECs) obtained from Tibetan and Han. METHODS: UVECs were obtained from native Tibetan and immigrant Han, respectively and cultured under hypoxia conditions (0.5% oxygen) for 2 h, 4 h, 12 h, and 24 h and normoxic conditions. VEGF, iNOS and eNOS mRNAs were detected with methods of RT-PCT. RESULTS: VEGF and iNOS mRNAs were up-regulated while eNOS mRNA depressed by hypoxia similarly in Tibetan and Han UVECs. CONCLUSION: Our results suggest that the changes of VEGF, iNOS and eNOS mRNA expression are common pathways in the mechanisms of hypoxic responses.
ABSTRACT
Thirty-seven cases of brain glioma,confirmed by light microscopy and immuno-'histochemistry were studied with transmission electron microscopy.It was found that there were certain ultrastructural features for each type and each grade of glioma.In astrocytoma,there was certain amount of glial filaments in the tumor cells; Rosenthal fibers consisted of irregular osmiophilic masses surrounded with glial filaments,occasionally cytoplasmic annulate lamellae and intranuclear filaments could be seen,and the interstitial capillaries were characteristic.In typical oligodendroglioma,astrocytic processes containing glial filaments were quite present in different amounts.These findings suggest that observation on the ultrastructure of gliomas is of significance to establish the diagnosis,to assess the degree of differentiation,and to identify some rare structures which can reveal the essence of the tumor.In addition,ultrastructural observation is helpful for prognosis.
ABSTRACT
This paper is to report the pathological changes in canine renal tissues after severe steam inhalation injuries. The specimens from 84 male mongrel dogs are studied with a light microscope, and 30 tissue samples of the 84 with an electron microscope. The morphological changes of glomeruli are characterized by, hypertrophy and/or hyperplasia of the glomerular cells, the former manifested as cell enlargement, increased amount of cytoplasm and rich in organelles; retrogressive changes of the glomerular cells in varying degrees as evidenced by intra-cellular edema. The renal tubules show varying degrees of degeneration, necrosis, and casts formation. Tubular necrosis affects more frequently the proximal convoluted tubules. The etiological factors of it are briefly discussed.
ABSTRACT
AIM: To explore the expression of CD14 in rat Kupffer cells (KCs). METHODS: In rat KCs induced by LPS or the mediators from KCs induced by LPS,the changes of CD14 expression were measured by RT-PCR and immunohistochemistry.The expressions of TNF? mRNA?IL-6 mRNA or the concentrations of TNF??IL-6 were estimated by in situ hybridization and radioimmunoassay,respectively. RESULTS: LPS increased the expression of CD14 in KCs in a dose-dependent fashion (LPS,1 ?g/L-10 mg/L) and in a time-dependent fashion(0.5 h-24 h,peaked at 3-6 hours). While the expression of CD14 in KCs stimulated by the active mediators from KCs which had been exposed to LPS 1 hour were obviously increased. CONCLUSIONS: There was a close relationship between LPS or the active mediators from KCs induced by LPS and the expressions of CD14. It is implied that the increase in CD14 expression may be induced by LPS and the cytokines produced by KCs,it also reveals that there is a auto-regulated loop in CD14 expression.
ABSTRACT
Objective To analyze the genetic quality and to determine the strain of laboratory mice. Methods Ten strains of mice were analyzed by using AFLP method. Results Polymorphism was detected in 10 strains of mice by 17 single enzyme primers and 20 pairs of double enzyme primers amplification. A total of 251 bands were shown by single enzyme AFLP in agarose gel with the size of the bands ranging from 100 bp to 2 000 bp and 89 polymorphic loci were detected. A total of 1507 clear bands between 50 bp and 600 bp were shown by double enzyme AFLP and 378 polymorphic loci were detected. Through statistical analysis, we calculated the similar index and genetic distance index. Our results showed BALB/c and BALB/c nu had the closest relationship and KM had a closer relationship with TA2, BALB/c and BALB/c nu, while DBA/2 showed a distant relationship with T739, 615 and C57BL/6J, coinciding with the origins of breeds. Conclusion Each strain could be distinguished from others by using the AFLP polymorphic primers, which provides reference data for genetic quality analysis and strain determination of mice.
ABSTRACT
Thirty-six cases of astrocytoma (10 cases each of gradeⅠ and grade Ⅱ,and eight cases each of grade Ⅲ and grade Ⅳ) with definite followup data were studied with microspec-trophotometry on Feulgen stained slides to determine the nuclear area and the DNA content.10 specimens of normal brain tissue or brain tissue with gliosis were observed with the same technique.It was found that there was no significant difference of the nuclear size between the cells of grade Ⅰ astrocytoma and those with gliosis,but the DNA content was significantly higher in the former than in the latter.And there was also defference of the histograms between the 2.The higher the grading of astrocytoma,the larger the nuclear area,and the higher the DNA content.The peak values of the histograms were shifted rightward with a scattering of the values.In addition,along with the increase of DNA index,there was a decline of the survival curve of the patients with an obvious shortening of the survival time.The findings suggest that nuclear DNA quantitation can serve as an auxiliary tool to differentiate grade.I astrocytoma from gliosis,to grade astrocytomas and to predict the prognosis.
ABSTRACT
Sixty-six rabbits were divided into 2 groups, the control group and the experimental group. The latter was subdivided into 10 groups according to the time of observation after burn injury including 2nd-hour group to 30th-day group. Each group consisted of 6 animals. Specimens from the trachea and the lungs were examined with optical microscopy, scanning electron microscopy and transmission electron microscopy.No obvious lesion was seen in the specimens from the control. In the experimental group, various pathological changes began to appear from the 6th hour after injury. In the trachea and bronchi, congestion of varying degrees, edema, leucocytic infiltration, lodging, adhesion, breaking or separation of cilia, and increase of goblet cells and Clara cells in number weie found. In. the lungs, interstitial edema of varying degrees, accumulation and infiltration of neutro-phils in capillaries, pulmonary interstitium and alveolar spaces, decrease in num ber of type II pneumocytes and their lamellar bodies, vacuolization of lamellar bodies, and phagocytosis of lamellar bodies by macrophages were seen. Most prominent changes were shown on the 3rd day postburn, and they began to alleviate on the 7th day. The number of type II pneumocytes and their lamellar bodies gradually increased number. Some lesions still existed on the 30th day postburn but no significant fibrosis could be found. The occurrence and development of the main lesions and their significance were discussed.