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The purpose of this review is to evaluate the promoting effects of amniotic membrane tissue on ulcer healing and investigate the underlying mechanism, providing new ideas for diabetic foot management. A computer-based online search of CNKI, PubMed and other databases to screen multi-center, randomized controlled trials published in high-impact journals using biological dressings, diabetic foot and other search terms. The retrieved data were analyzed and summarized. Amniotic membrane tissue and its derivatives can greatly shorten the time taken for healing of ulcer surface. This occurs possibly because they can promote angiogenesis and neural repair and prevent against infection. Moreover, the new derivatives provide great ease in use. Therefore, amniotic membrane and its derivatives provide a new tool for diabetic foot management.
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Objective To investigate the effects of sodium butyrate on ethanol-seeking behavior and H3K9 acetylation levels in NMDA receptor 2B subunit(NR2B) promoter region in the hippocampus of Wistar rats.To explore the epigenetic mechanism underlying ethanol-seeking behavior.Methods According to random number table,48 male Wistar rats were divided into saline group,sodium butyrate group,ethanol group and sodium butyrate + ethanol group,with 12 rats in each group and administered by intraperitioneal injection respectively.Conditioned place preference (CPP)was used to evaluate the ethanol-seeking behavior.Using Western-blot,real-time PCR and chromatin immunoprecipitation assays,the expression of NR2B protein,NR2BmRNA and the relative level acetylated H3K9 in NR2B promoter region in the hippocampus were determined respectively.Results The CPP test and the CPP score in each group were different (P＜ 0.05).Compared with the CPP test(261.1 ± 102.2) and the CPP score(48.5±94.6) of saline group,the CPP test ((406.8±109.2),(502.7±72.89)) and the CPP score((198.2± 119.4),(277.5±76.2)) of ethanol group and sodium butyrate + ethanol group were significantly higher(P＜0.05),the CPP test(193.4±93.8) and the CPP score (9.7±94.0)of sodium butyrate group were not significantly different(P＞0.05).Compared with the ethanol group,CPP test of sodium butyrate + ethanol group was significantly higher(P＜0.05).The expression of NR2B protein,NR2BmRNA and acetylated level H3K9 in NR2B promoter region in the hippocampus in each group were different (P＜ 0.05).Compared with the expression of NR2B protein (1.00 ± 0.28),NR2BmRNA(1.00±0.14) and H3K9 acetylation in NR2B promoter region(1.00±0.25)in the hippocampus of saline group the expression of NR2B protein((1.40±0.34),(1.79±0.30)),NR2BmRNA((1.26±0.16),(1.50±0.08)) and aeetylated level H3K9 in NR2B promoter region ((1.68±0.16),(2.35±0.45)) of ethanol group and sodium butyrate ± ethanol group were significantly higher(P＜0.05).The expression of NR2B protein(0.85±0.24),NR2BmRNA(1.05±0.13) and acetylated level H3K9 in NR2B promoter region(0.96±0.41) of sodium butyrate group were not significantly different(P＞0.05).Compared with the ethanol group,the expression of NR2B protein,NR2BmRNA and acetylated level H3K9 in NR2B promoter region in the hippocampus of ethanol group,these of sodium butyrate + ethanol group were significantly higher (P＜0.05).The CPP score were positively correlated with the expression of NR2B protein (r=0.474,P＜0.05).The expression of NR2B protein were positively correlated with the expression of NR2BmRNA (r=0.468,P＜0.05).The expression of NR2BmRNA were positively correlated with the expression of H3K9 acetylation in NR2B promoter region(r=0.596,P＜0.05),and the CPP score were positively correlated with the expression of H3K9 acetylation in NR2B promoter region (r=0.542,P＜0.05).Conclusion The increasing acetylation level of H3K9 in NR2B promoter region in the hippocampus may be one of the epigenetic mechanisms of promoting ethanolseeking behavior,and H3K9 deacetylation in NR2B promoter region in the hippocampus is likely to be a new target for controlling ethanol dependence.
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Objective To investigate the therapeutic effect of self made balloon with side hole on no reflow ( NR) after emergency percutaneous coronary intervention ( PCI ) .Methods 48 patients with NR after PCI in our hospital were randomized into two groups , which were group A ( n=24 , patients received self made perfusion balloons with holes ) and group B ( n=24 , patients using direct guiding catheter ) and through respective devices intravascular tirofiban and verapamil were given .TIMI flow grade, recovery of myocardial enzymes and ST-segment elevation , LVEF and the incidence of MACE were compared between the 2 groups.Results Among patients in group A , the percentage of immediate postoperative TIMI Ⅲflow (79.2%vs.45.8%,P=0.032), ST segment resolution of more than 50% (83.3% vs.54.2%,P =0.029 ) and LVEF after 1 months [ ( 54.92 ±12.32 )% vs. ( 47.67 ±12.15 )%, P =0.046 ] were significantly higher than patients in group B .The CK peak value of patients in group A [ ( 1018.62 ± 732.34)mmol/L vs.(1497.75 ±858.63)mmol/L, P =0.043], CK-MB peak values [(113.84 ± 76.53 ) mmol/L vs.( 172.74 ±93.56 ) mmol/L, P=0.021 ] and MACE rates ( 0 vs.16.7%, P=0.037 ) were lower than those of patients in group B .Conclusions The use of self-made perfusion balloon with side hole for the treatment of NR patients after emergency PCI is convenient , easy and effective.
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Objective To observe the changes of N-methyl-D-aspartate (NMDA) receptor 2B subunit (NR2B) expression in the striatum of chronic alcohol exposured rats at different withdrawal time.Methods 72 male Wistar rats were randomly divided into withdrawal 2h group,withdrawal 6h group,withdrawal 12h group,withdrawal 1d group,withdrawal 3d group and control group,and 12 rats in each group.In the 5 withdrawal groups,ethanol was administered in drinking water at the concentration of 6％ (V/V) for 16 weeks,and rats in control group were maintained with water.After 16 weeks ethanol was removed and ethanol withdrawal syndromes were evaluated.The expression of NR2B protein in the striatum was measured by immunofluorescence and western blot and the expression of NR2B mRNA in the striatum was measured by realtime PCR.Results Compared with withdrawal scores of control group((1.50±0.80)),scores of withdrawal 2h,6h,12h,1d,3d groups ((10.42±2.50),(15.42± 1.93),(9.25±2.01),(7.67± 1.92),(2.25±0.87) respectively) were higher,and the withdrawal scores of withdrawal 6h group were the highest.Compared with the expression of NR2B fluorescence intensity (2210.00± 178.20),the expression of NR2B protein(0.150±0.009) and the expression of NR2B mRNA(0.006±0.001) in the striatum of control group,the expression of NR2B fluorescence intensity (2710.56 ± 194.21),(5035.16 ± 234.41),(3326.23 ± 378.16),(2570.64 ±177.88),the expression of NR2B protein (0.192±0.008),(1.649±0.205),(0.783±0.109),(0.180±0.009) and the expression of NR2B mRNA (0.026±0.002),(0.351±0.034),(0.248± 0.023),(0.024±0.003) of withdrawal 2h,6h,12h,ld groups were significantly higher (P＜0.05),and with the extension of the withdrawal time,the expression was gradually increased.The expression of withdrawal 6h group was the highest,then began to decline,and returned to baseline levels at withdrawal 3 d(P＞0.05).Withdrawal scores were positively correlated with the expression of NR2B protein(r=0.719,P＜0.01),the expression of NR2B protein was positively correlated with the expression of NR2B mRNA(r=0.937,P＜0.01),and the expression of NR2B mRNA was positively correlated with withdrawal scores(r=0.673,P＜0.01).Conclusion The expression of NR2B was up-regulated in the striatum of chronic alcohol exposured rats at different withdrawl time.NR2B protein and NR2B mRNA expression is positively correlated with the withdrawal scores,suggesting that regulating the expression of NR2B may be a new target for the treatment of ethanol withdrawal symptoms.
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Objective To explore the effect of tumor necrosis factor-alpha(TNF-α) blockade therapy on circulating Th17 cell percentage and serum interleukin (IL)-17 level in patients with rheumatoid arthritis (RA) and ankylosing spondylitis (AS). Methods Twenty-seven RA and 22 AS patients were recruited, of which 14 cases from both diseases received 40 weeks TNF blockade therapy. Twenty-four healthy blood donors were used as controls. The frequencies of circulating Th17 cells were determined by flowcytometry, and serum IL-17 level were measured by enzyme linked immunosorbent assay(ELISA). Results Significantly higher baseline circulating Th17 cells were observed in active RA and AS patients compared with the healthy controls[RA 1.03%(0.66%,1.78%) vs controls 0.50%(0.43%,0.67%), Z=-3.236, P<0.01; AS(1.16±0.09)%vs controls (0.59 ±0.061)% , t =5.226, P <0.01]. Similarly, serum IL-17 level were significantly elevated in patients with both diseases compared with controls[RA(32.3±2.5) pg/ml vs controls(14.3±2.5) pg/ml, t=5.070, P<0.01; AS 28.98(23.84,36.14) pg/ml vs controls 11.84(5.33,22.12) pg/ml, Z=-4.103, P<0.01]. After TNF-α blockade therapy, serum IL-17 was significantly decreased in both diseases groups[RA △(-13.5± 5.0) pg/ml and AS △(-16.0±1.9) pg/ml]. In contrast, no significant differences were found in the frequencies of circulating Th17 cells[RA △(0.104 5±0.212 6)% and AS △(0.002 5±0.183 8)%]. Conclusion Th17 cells and IL-17 have been implicated in the pathogenesis of RA and AS. TNF-α blockade can partially inhibit the function of Th17 cells. However, it is unable to reduce the frequencies of these cells in the circulation after 40 weeks therapy, which may explain the reasons for the relapse.
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Objective To study the serum levels of low molecule weight IgM (LMW IgM) in ankylosing spondylitis (AS) patients and rheumatoid arthritis (RA) patients and to evaluate the relationship of LMW IgM levels with the disease activities. Methods The levels of LMW IgM and pentameric IgM in AS patients, RA patients and healthy controls were measured with ELISA after separated using ultrafiltration assay. Differences in the percentage of LMW IgM between subject groups were analysed using Mann-Whitney U test. Results The percentages of LMW IgM increased dramatically in AS patients and RA patients compared with healthy controls (0.194 ± 0123, 0.061 ±0.026, 0.028 ±0.165 separately). The LMW IgM percentages were not correlated with the disease activities. Conclusion The increase of LMW IgM indicates humoral immune function abnormality in AS patients and RA. However, the mechanism needs further study.
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Background and purpose: Infiltration and metastasis are characteristic of the biological behavior of cancer. Blood circulation and lymphatic spread are two important ways for neoplasm metastasis. The lymphatic vessel is one of the earliest routes for solid neoplasm metastasis. However, compared to tumor blood vessels, there were only a few studies on the research for lymphatic vessel spread. In recent years, with the identification of vascular endothelial growth factor-C (VEGF-C), VEGF-D and lymphatic endothelial markers including lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), vascular endothelial growth factor receptor-3 (VEGFR-3), glomerular podocyte membrance mucoprotein (podoplanin) and the homeobox transcription factor (Prox-1), lymphangiogenesis has become one of the important areas in the study of tumor metastasis. This paper was to study the expression of proprotein convertase (PC)5, PC7, VEGF-C, VEGF-D and their receptor VEGFR-3 in patients with non-small cell lung cancer (NSCLC) and their clinico-pathoiogical value. Methods: Twenty specimens of the NSCLC, peritumoral tissues as experimental group and nine pulmonary benign diseases as control group were studied. The expression of PC5, PC7, VEGF-C, VEGF-D and VEGFR-3 mRNA in specimens of those tissues were studied by real-time quantitative reverse transcriptase polymerase chain reaction (real-time quantitative RT-PCR). Results: ①The expressions of PC5, PC7, VEGF-C, VEGF-D, VEGFR-3 mRNA in specimens of NSCLC were significantly higher than those of the peritumoral and pulmonary benign diseases tissues (P<O.05).② The expressions of PC5, PC7, VEGF-C, VEGF-D and VEGFR-3 mRNA in NSCLC were not correlated with age, gender, site and dimension of lesion, types of histological and degree of differentiation, but correlated significantly with lymph node metastasis (P=0.000, P=0.000,P=0.012,P=O.000, P=0.004) and pTNM stage (P=0.011, P=0.012, P=0.O13, P=0.011, P=0.028).③Significant correlationsbetween PC5 and VEGF-C (r=0.461, P=0.041), PC5 and VEGF-D (r=0.793, P=0.000), PC5 and VEGFR-3 (r=0.498, P=0.026), PC7 and VEGF-C (r=0.450, P=0.047), PC7 and VEGF-D (r=0.699, P=0.001), PC7 and VEGFR-3 (r=0.616, P=0.004), VEGF-C and VEGF-D (r=0.532, P=0.016), VEGF-C and VEGFR-3 (r=0.607, P=0.001), VEGF-D and VEGFR-3 (r=0.451, P=0.048) mRNA were observed in NSCLC. Conclusion: Non-small cell lung cancer cells may secrete lymphangiogenetic growth factors like VEGF-C, VEGF-D and their receptor VEGFR-3 and may associate with lymphatic metastasis of NSCLC through their downstream pathways. The expression of VEGF-C, VEGF-D, VEGFR-3, PC5 and PC7 may serve as important markers for evaluation of lymphatic metastasis and prognosis in patients with NSCLC.
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Background and purpose:Inf iltration and metastasis are characteristic of the biological behavior of cancer.Blood circulation and lymphatic spread are two important ways for neoplasm metastasis.The lymphatic vessel is one of the earliest routes for solid neoplasm metastasis.However,compared to tumor blood vessels,there were only a few studies on the research for lymphatic vessel spread.In recent years,with the identifi cation of vascular endothelial growth factor-C(VEGF-C),VEGF-D and lymphatic endothelial markers including lymphatic vessel endothelial hyaluronan receptor-1(LYVE-1),vascular endothelial growth factor receptor-3(VEGFR-3),glomerular podocyte membrance mucoprotein(podoplanin)and the homeobox transcription factor(Prox-1),lymphangiogenesis has become one of the important areas in the study of tumor metastasis.This paper was to study the expression of proprotein convertase(PC)5,PC7,VEGF-C,VEGF-D and their receptor VEGFR-3 in patients with non-small cell lung cancer(NSCLC)and their clinico-pathological value.Methods:Twenty specimens of the NSCLC,peritumoral tissues as experimental group and nine pulmonary benign diseases as control group were studied.The expression of PC5,PC7,VEGF-C,VEGF-D and VEGFR-3 mRNA in specimens of those tissues were studied by real-time quantitative reverse transcriptase polymerase chain reaction(real-time quantitative RT-PCR).Results:①The expressions of PC5,PC7,VEGF-C,VEGF-D,VEGFR-3 mRNA in specimens of NSCLC were signifi cantly higher than those of the peritumoral and pulmonary benign diseases tissues(P
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Background and purpose:Conventional cytology is valuable in diagnosing the cancer cells in lymph nodes of patients with lung cancer. However , the diagnostic value of detecting lymph node and paracancer micrometastasis was limited. Our study was to investigate the mRNA expression level of lung specif ic X protein(LUNX) and cytokeratin 19(CK19) and its signifi cance in the carcinogenesis,metastasis and prognosis of NSCLC. Methods: Quantitative analysis with reverse transcriptase polymerase chain reaction(Real-Time RT-PCR) was used to detect the mRNA expression level of LUNX and CK19 in 20 tumor tissues and 42 regional lymph nodes from 20 patients with NSCLC, and 6 lymph nodes from 9 patients with pulmonary benign diseases as control. Meanwhile, all lymph nodes were also examined by conventional pathological method. Results:①the mRNA level of Lunxin in cancer Tissue was signifi cantly higher than that in the control group, but there was no association with Lymphatic Metastasis,Tumor Size and TNM grade.②the mRNA level of Lunx in the cancer Tissues was signif icantly higher than that in the control group, and was associated with Lymphatic Metastasis,Tumor Size and TNM grade.③the mRNA of CK19 in lymph nodes was almost the same as control, and not associated with tumor size,lymphatic metastasis and TNM grade.④the mRNA of LUNX in lymph nodes was almost the same as control, which has no acossiation with lymphatic metastasis,tumor size and TNM grade.⑤ mRNA of CK19 and LUNX was unrelated to age,sex and histological type .⑥there was signifi cant difference between using Real-Time RT-PCR methods and the routine pathological methods to detect lymph node metastasis in lung cancer. Conclusion:This result suggested that the detection of LUNX message RNA and CK19message RNA might be helpful to diagnose NSCLC micrometastasis in lymph nodes. LUNX was superior to CK19 both in sensitivity and specifi city. The establishment of this method may increase the positive rate of detecting metastatic regional lymph nodes in non small cell lung cancer.
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AIM: To explore the histochemical changes o f diabetic skin and the pathogenesis of impaired wound healing in diabetes. METHODS: 54 male Sprague-Dawley (SD) rats weighing 200-220 g wer e randomized into control and STZ-induced diabetic groups. The shaved skin speci mens from the back of rats were collected in 4, 8 and 12 weeks post STZ-inductio n, respectively. Hematoxylin-eosin dye was used for histological examination. Me a nwhile, the skin glucose contents were measured by Beckman's autoanalyzer. Skin AGEs concentrations were assessed by detecting total fluorescence in tissue coll agen and immunohistochemistry assay. RESULTS: The skin thickness in diabetic animals was decreased, w ith the features of multilayer epithelium structure disappeared in epidermis and collagen fibers atrophied, swollen and degenerated in dermis; The inflammatory responses in the dermis of diabetic animals were increased obviously. The result s also revealed that skin glucose contents in diabetic rats [(2.64?1.03)mg/ g skin] were 2-3 times higher than those in the controls [(0.74?0.33)mg/g s kin] (P
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To observe the effects of melatonin on postacyclin concentration, nitric oxidase activity(NOS) and inducible nitric oxidase(iNOS) mRNA expression in calf aortic endothelial cells in medium with high concentration of glucose, endothelial cells of calf thoracic arota were cultured and divided into five groups. Group A was a low-glucose control with 5.5mmol/L glucose in medium. Group B was a high-glucose control with 33mmol/L glucose in medium. Cells of group C was cultured with 33mmol/L glucose and 10 -13mol/L melatonin, those of group D was cultured with 33mmol/L glucose and 10 -9mol/L melatonin, and those of group E was cultured with 33mmol/L glucose and 10 -5mol/L melatonin. The concentration of 6-keto-PGF 1?, metabolizing product of PGI 2, in the culture medium was measured with radio-immunological assay. NOS activity was determined with colorimetry and iNOS mRNA was measured with semi-quantitative RT-PCR. 6-keto-PGF 1? concentration, NOS activity and iNOS mRNA expression in group A were significantly lower than those in group B(P0.05). 6-keto-PGF 1? concentration of group D and group E was significantly higher than that in group B(P
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To investigate the protective effect of melatonin on the kidneys of STZ-induced diabetic rats, experimental diabetes was induced by intraperitoneal injection of streptozotocin 50 mg/kg in male Sprague-Dawley rats (150～200g). Three days after streptozotocin injection, the diabetic rats were assigned to one of the following groups: (1) DM, untreated; (2) DM+Mel 1, melatonin supplement at 0.2mg?kg -1?d -1, by gavage; (3) DM+Mel 2, melatonin supplement at 5mg?kg -1?d -1, by gavage. The treatment continued for 8 weeks. Periodically, body weight, blood glucose, plasma triglyceride and cholesterol levels were measured as clinical and biochemical parameters; plasma creatinine, creatinine clearance( Ccr), urinary protein excretion were detected as renal function indexes; kidney weight, glomerular area, PAS positive area, and cell counts were evaluated as histological damage indexes. Melatonin treatment had no significant effects on blood glucose and plasma cholesteral, but lowered plasma triglyceride level in diabetic rats. Melatonin treatment reduced urinary protein excretion. Until the end of the experiment, there were no significant changes in plasma creatinine level, Ccr in diabetic rats. Melatonin treatment decreased kidney weight/body weight ratio, reduced PAS positive area, glomerular cellular number and glomerular area which were increased in DM rats. The protective effects of melatonin were more obviose in high dosage groups. Melatonin treatment can aggravate glomerular hypertrophy and mesangial cell proliferation, delay the progression of glomerulosclerosis, and improve renal function of diabetic rats.