1.Expression of autophagy related proteins in neonatal rats after early infection induced celebral matter damage
Meng MENG ; Xingxing LU ; Yan XU ; Jun WANG
Chinese Journal of Immunology 2017;33(8):1152-1155,1160
Objective:To explore the effect of infection on autophagy-related proteins,Beclin-1 and LC3,expression in cerebral white matter in newborn rats.Methods: A total of 64 two-day-old Sprague-Dawley(SD) rats were randomly divided into control and experimental groups(n=32 each).At day 2 to 6 after birth,the rats in experimental group were intraperitoneally injected with 0.6 mg/kg of lipopolysaccharide(LPS) once a day to establish a white matter injury induced by infection in neonatal rats while the rats in control group were injected with equal amounts of normal saline.Rats were sacrificed to collect brain tissues at 12 hour,1,3,5 d after model establishment.HE staining was performed to observe the pathological changes.Changes in the expression of Beclin-1 and LC3 in rat white matter were examined by Western blot and RT-PCR.Results: Growth and development of rat in experimental group was slow,cerebral white matter lesions were obvious.Compared with the control group,the experimental group Beclin1 and LC3 protein and mRNA levels in the model after 12 h began to express,1 d reached the peak,and then decreased,each time points were higher than the control group(P<0.05).Conclusion: Early infection in neonatal rats can cause white matter damage;the expression of autophagy-related proteins,Beclin1 and LC3,showed that autophagy may be involved in the pathological process of white matter damage induced by infection.
3.Study on transplantation of induced bone marrow mesenchymal stem cells via various route for the treatment of chronic liver injury
Yan SUN ; Baorong CHI ; Lu CHEN ; Xiangwei MENG ; Dexia KONG
Chinese Journal of Digestion 2008;28(3):171-174
Objective To investigate the efficacy of transplantation of induced bone marrow mesenchymal stem cells(MSCs)via various route for treatment of chronic liver injury.Methods MSCs were isolated and expanded by density gradient centrifugation combined with adherent culture.MSCs were induced to differentiate into hepatocyte-like cells in vitro.The markers of hepatocyte lineage were examined by RT-PCR and immunocytochemistry.The changes of ultramicrostructure of MSCs were observed by transmission electron microscope.One hundred and fourteen Wistar rats were treated with mixture of 40%CCl4 and peanut oil to establish chronic liver injury model and another 6 rats were served as control.Twelve rats in model group were died within 8 weeks,and the rest rats were divided into portal vein,spleen and tail vein transplantation groups.The rats were transplanted with DAPI-labelled MSCs which were induced for 14 days.The distribution of the DAPI-labelled cells were observed by fluorescence microscopy.The liver function and pathology were examined.Results AFP mRNA expression were detected at 7th and 14th day after induction,expressions of ALB mRNA,CK18 and hepatocyte antigen were found at 14th and 2lth day.The ultrastructure examination revealed that MSCs were enlarged with a lot of endoplasmic reticulum,mitochondrion,lysosome and glycogen granule.DAPI labelled cells were found in the spleen and liver transplantion groups.The pathological changes of liver were alleviated in all treated groups,especially in intraspleenic transplantion group(P<0.05).Conclusions Induced bone marrow MSCs can ameliorate liver function of chronic hepatic injury after transplantation.The intraspleenic transplantation is better than others.
5.Application of the modulation transfer function to evaluate the early visual quality after full femtosecond laser refractive surgery
Hu, MENG ; Yan, LU ; Peili, HOU ; Zhenping, HUANG
Chinese Journal of Experimental Ophthalmology 2014;32(5):420-424
Background Full femtosecond laser refractive surgery has been generally applied,but the improvement of visual quality after surgery is a vital problem.It is well-known that modulation transfer function (MTF) is a major evaluation indicator.Objective To investigate the change of the MTF of operated eyes after the full femtosecond laser refractive surgery.Methods A retrospective series of cases study was adopted.Forty eyes of 23 patients with the spherical equivalent of-2.25 to-5.75 D who received full femtosecond laser refractive surgery were enrolled in Nanjing General Hospital of Nanjing Military Command from July to October,2012.The change of visual acuities was analyzed and compared among before operation and 1 day,1 week and 1 month after operation.MTF value in total aberration,low and high order aberrations under the 3.0 mm and 6.0 mm pupil size conditions were detected with i-Trace Visual Function Analyzer at above time points.The MTF values among various time points,between different pupil diameters and different spatial frequencies (5,10,15,20,25 c/d) were compared respectively.Results Compared with the preoperation,the visual acuity improved in postoperative 1 day,1 week and 1 month,showing a significant improvement among various time points (F =10.341,P =0.000).Corrected diopter was 3.795 D at average.MTF values under the 3 mm pupil diameter gradually elevated with the lapse of time after operation,without significant differences among different time points whatever total aberration,low and high order aberrations(all at P>0.05).The MTF values under 6 mm pupil diameter of total aberration and low order aberration were significantly higher in postoperation than those in preoperation(all at P<0.05),but no significant difference was seen in MTF of high order aberration among various time points (F =0.260,P =0.854).Compared with the MTF under the 3 mm pupil diameter,those of low and high order aberrations under the 6 mm pupil diameter considerably declined in preoperation and postoperative 1 month (preoperation:P =0.050,0.001 ; postoperation:P =0.012,0.001).MTF values under the 3 mm pupil diameter were not different at various spatial frequencies (all at P> 0.05).However,MTF values under the 6 mm pupil diameter were gradually increased from preoperation through postoperative 1 month at 10,15,20 and 25 c/d (P =0.044,0.043,0.024,0.014).Conclusions Full femtosecond laser refractive surgery can eliminate low aberration,increase the MTF and acuity,and therefore improve the visual quality.MTF is one of the key indicators assessing visual quality after full femtosecond laser refractive surgery.
6.Inhibition of invasive and chemotactic abilities of SKOV3 cells bv human epithelial growth receptor-2 small interfering RNA
Shu-Lan ZHANG ; Yan-Ming LU ; Li-Rong MENG ; Yan-Yan ZHAO ;
Chinese Journal of Obstetrics and Gynecology 2001;0(01):-
Objective To investigate the effects of RNA interference(RNAi)targeting human epithelial growth receptor-2(HER-2)gene on the invasive and chemotactic ability of SKOV3 cells. Methods Glyeeraldehyde-3-phosphate dehydrogenase(GAPDH)was used as the positive control. Lipofectamine 2000 mediated transient transfection was conducted to transmit the siRNA into SKOV3 cells. Three pairs of specifically targeted(HER-2 siRNA Ⅰ,HER-2 siRNA Ⅱ,HER-2 siRNA Ⅲ)sequence were selected in the coding region of HER-2 mRNA.Transfection of HER-2 siRNA was conducted with lipofeetamine 2000 in ovarian carcinoma cell line SKOV3.The HER-2 gene expression was assessed by real- time PCR and western blot assays.Changes of invasive and chemotaetie capacity of SKOV3 cells were measured by polycarbonates coated with or without matrigal.Results Western blot results showed that the expression of GAPDH protein was decreased in specifically transfected cells and with the increase of siRNA dose,the expression of GAPDH protein was decreased.GAPDH protein gray value in control group, different doses(0.5,1.0,1.5,2.0 ?g)GAPDH siRNA interference groups were 0.6855?0.0259,0.5698 ?0.0275,0.4542?0.0296,0.3341?0.0178 and 0.1816+0.0180,respectively.There was a significant difference in each group(F=198.126,P
7.The expression of V-ATPase and its significance in colon cancer
Senlin ZHAO ; Bin MENG ; Dongwang YAN ; Likang ZHANG ; Feifei CUI ; Huijun LU ; Jian CHEN ; Zhihai PENG
Journal of Regional Anatomy and Operative Surgery 2014;(5):486-488
Objective To explore the expression of V-ATPase in colon cancer and its clinical significance. Methods Detecting the expression of V-ATPase mRNA in 20 paired of colon tumor tissues and normal tissues by using reverse transcription-polymerase chain reaction ( RT-PCR) and real-time quantitative polymerase chain reaction( Real-time PCR) , and testing the expression of V-ATPase protein by immu-nohistochemistry of EnVinsion. Results The expression of V-ATPase mRNA in tumor tissues and its paired normal tissues were (5. 37 ± 0. 44) and (2. 03 ± 0. 35)(P<0. 01). The positive immunohistochemistry of V-ATPase in tumor tissues and its paired normal tissues were 69. 1%(47/68) and 5. 8%(4/68) respectively, and the positive expression were primarily in cytoplasm and cytomembrane. Overexpression of V-ATPase was associated with tumor stage (P<0. 05), lymph node metastasis (P=0. 044), distant metastasis (P=0. 049), vessel in-vasion (P=0. 044) and differentiation (P<0. 001). Conclusion Overexpression of V-ATPase plays a significant role in the carcinogene-sis and the progression of colon cancer, which might be an important postoperative therapeutic target.
8.Clinical Traits and Treatment of Urinary Tract Infection Based on TCM Symptom Differentiation among Elderly Patients
Min YU ; Jianjun LU ; Weihui LI ; Yan HE ; Meng LI ; Yaoxun SHI ; Anna WANG
Chinese Journal of Nosocomiology 2006;0(09):-
OBJECTIVE To discuss the clinical traits,pathogenesis and TCM stepwise treatments of urinary tract infection(UTI) based on symptom differentiation among elderly patients.METHODS Because of the whole and the partial immune defense function descending,the elderly patients easily suffered from UTI,which was characterized by non-typical symptom,complex and serious condition.In the aspects of the pathogenesis of UTI among elderly patients,the principal aspect of which was the deficiency of the kidney and spleen(weakened body resistance)and the secondary incidental was accumulated damp-heat in the lower warmer(affected pathogenic factor),viz weaken healthy qi and excessive pathogenic factor.Hence during the clinical treatment we should attach importance to regulate entire faculty condition.Considering the acute attack stage and non-acute phase,we will differently inflict therapies of clearing away the heat-evil,dissolving dampness,treating stranguria,hemostasis,and invigorating spleen and kidney assisted by treating stranguria and so on,according to differentiation of symptoms and treatments,in which way we could give attention to both the principal and the secondary aspects of a disease.RESULTS The TCM stepwise therapy of UTI among elderly patients had the characteristics of high efficacy,few side reactions and stable long-term curative effects.CONCLUSIONS The TCM therapy of UTI among elderly patients has more potentiality and predominance,which deserves further study.
9.Effects of Herb-partition Moxibustion at Navel on Protein and mRNA Expressions of Bcl-2 and Bax in Rats Model with Premature Ovarian Failure
Yan LU ; Meng TIAN ; Qingsi SHEN ; Jing ZHANG ; Zhengao SUN ; Yongchen ZHANG ; Hongling JIA
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(7):40-44
Objective To observe the effects of herb-partition moxibustion at navel on the expressions of Bcl-2 and Bax in rats with premature ovarian failure (POF); To explore its mechanism underlying improvement of POF. Methods POF model was established by intraperitoneal injection of cyclophosphamide. Forty-eight Female Wistar rats were randomly divided into normal group, model group, moxibustion group and Western medicine group, with 12 rats in each group. Rats of moxibustion group were treated with herb-partition moxibustion at navel (Shenque, CV8), 30 min/d, once every other day; rats of Western medicine group were given intragastric dose of estradiol valerate, once a day, for 14 d. The protein and mRNA expressions of Bax and Bcl-2 in ovarian granulosa cells were detected by immunohistochemal method, Western blot and qRT-PCR, respectively.Results The weights of ovaries in the model group rats were significantly lower than those in the normal group (P<0.05). The expressions of Bcl-2 and Bax protein in ovarian tissue of each group were mainly expressed brown coloration and in the cytoplasm of cells. The range of Bax protein in granulosa cells of moxibustion group and Western medicine group was significantly less than that in model group. The range of Bcl-2 in moxibustion group and Western medicine group was significantly larger than that in model group. Compared with the normal group, the expressions of Bcl-2 protein and mRNA in the model group rats significantly decreased (P<0.05,P<0.01), and the expression of Bax protein and mRNA significantly increased (P<0.05,P<0.01). The expression level of Bcl-2 protein and mRNA was significantly higher in the moxibustion group and in the western medicine group than that in the model group (P<0.05,P<0.01). The expression level of Bax protein and mRNA in the moxibustion group was significantly lower than that in the model group (P<0.05,P<0.01). ConclusionThe herb-partition moxibustion at navel can improve the function of ovaries by up regulating the expression of Bcl-2 and down regulating the expression of Bax in POF rats model.
10.In vitro anti-inflammatory and free radical scavenging activities of flavans from Ilex centrochinensis.
Lu-jun LI ; Li-juan YU ; Yan-ci LI ; Meng-yuan LIU ; Zheng-zhi WU
China Journal of Chinese Materia Medica 2015;40(8):1523-1528
This study was carried out to evaluate the anti-inflammatory and free radical scavenging activities of flavans from flex centrochinensis S. Y. Hu in vitro and their structure-activity relationship. LPS-stimulated RAW 264.7 macrophage was used as inflammatory model. MTT assay for cell availability, Griess reaction for nitric oxide (NO) production, the content of TNF-alpha, IL-1beta, IL-6 and PGE, were detected with ELISA kits; DPPH, superoxide anion and hydroxyl free radicals scavenging activities were also investigated. According to the result, all flavans tested exhibited anti-inflammatory effect in different levels. Among them, compounds 1, 3, 4 and 6 showed potent anti-inflammatory effect through the inhibition of NO, TNF-alpha, IL-lp and IL-6, of which 1 was the most effective inhibitor, however, 2 and 5 were relatively weak or inactive. The order of free radical scavenging activities was similar to that of anti-inflammatory activities. Therefore, these results suggest that 3, 4 and 6, especially of 1, were,in part responsible for the anti-inflammatory and free radical scavenging activity of Ilex centrochinensis. Hydroxyl group at 4'-position of B-ring plays an important role in the anti-inflammatory and free radical scavenging capacities.
Animals
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Anti-Inflammatory Agents, Non-Steroidal
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chemistry
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pharmacology
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Cell Line
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Cyclooxygenase 2
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immunology
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Drugs, Chinese Herbal
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chemistry
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pharmacology
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Flavanones
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chemistry
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pharmacology
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Free Radical Scavengers
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chemistry
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pharmacology
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Ilex
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chemistry
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Interleukin-6
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immunology
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Macrophages
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drug effects
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immunology
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Mice
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Nitric Oxide
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immunology
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Tumor Necrosis Factor-alpha
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immunology