1.Effects of Lu Jiao Fang on Myocardial Fibrosis and Plasma Aldosterone Content in the Rat with Increase of PressureLoading
Hui CAI ; Wanying HU ; Yaorong DONG
Journal of Traditional Chinese Medicine 1993;0(06):-
Effects of Lu Jiao Fang on myocardial fibrosis and plasma aldosterone (ALD) content in the rat with increase of pressure - loading were investigated for approach to the machanism of anti - myocardial fibrosis. Left ventricular mass index(LVMI), myocardial collagen Ⅰ, Ⅱ Ⅲ and plasma ALD content were observed. Results showed that LVMI in the model group was significantly higher than that in the sham operation group(P
3.Expression,Purification,Preparation of Polyclonal Antibody and Subcelluar Localization of the NS2 Protein of Periplaneta fuliginosa Densovirus
Bo YANG ; Peiran YU ; Dawei CAI ; Xiaomin DONG ; Zhigang LIU ; Zheng HU ; Jiamin ZHANG ; Yuanyang HU
Microbiology 2008;0(09):-
NS2 is a nonstructural protein of Periplaneta fuliginosa densovirus(PfDNV) with a molecular mass of 30 kD,whose function is not yet clearly understood. In order to study the expression,subcellular distribution and the function of NS2 protein,the coding region of NS2 was amplified from the hindgut tissue of cockroaches infected with PfDNV by RT-PCR and then the recombinant prokaryotic expression vector pET28a-NS2 was constructed. The recombinant plasmid was transformed into E. coli BL21(DE3) to express the 6?His fusion protein in the bacteria. After purification,the fusion protein was injected into New Zealand rabbits to prepare polyclonal antibody. The specificity of the anti-NS2 antibody was successfullyproved by western blotting on the eukaryotic expressed products of NS2 protein.Meanwhile,the full sequence of ns2 gene was also cloned into the eukaryotic expression vector pAC. The recombinant plasmid pAC-NS2 was then transfected into Schneider line 2(S2) cells to express NS2 protein in the insect cells. The subcellular localization of NS2 in the insect cells was then investigated by indirect immunofluorescence technique using the anti-NS2 polyclonal antiserum. The confocal laser scanning microscope observation showed that NS2 protein was located primarily in the cytoplasm with some punctate nuclear staining.
4.Morphology and mechanical properties of normal lymphocyte and Jurkat revealed by atomic force microscopy.
Xiaofang CAI ; Jiye CAI ; Shisong DONG ; Hua DENG ; Mingqian HU
Chinese Journal of Biotechnology 2009;25(7):1107-1112
Alternations of lymphocyte in biophysical properties (e.g., morphology and viscoelasticity) are related to the human health, disease diagnosis and treatment. Here, we used atomic force microscopy (AFM) to characterize the morphology and mechanical properties of normal lymphocyte and Jurkat. The AFM images revealed that their cell shapes appeared similar. The mechanical properties of the two groups were tracked with AFM-based force spectroscopy. The normal lymphocyte cells had a high adhesion force distribution in (796.7 +/- 248.5) pN, whereas the Jurkat cells had a low force distribution in (158.5 +/- 37.5) pN. The adhesion force revealed that the Young's modulus of normal lymphocyte cells (0.471 kPa +/- 0.081 kPa) was nearly four times higher than that of Jurkat cells (0.0964 kPa +/- 0.0229 kPa) at the same loading rate. The stiffness of normal lymphocyte cells was (2.278 +/- 0.488) mN/m and that of Jurkat cells was (4.322 +/- 0.382) mN/m. The differences in mechanical properties of normal and cancerous cells were obvious that healthy and diseased states could be clearly distinguished. These results may be applied to the clinic disease diagnosis for distinguishing the normal cells from the cancer ones even when they show similar shapes.
Biomechanical Phenomena
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Humans
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Jurkat Cells
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physiology
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ultrastructure
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Lymphocytes
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physiology
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ultrastructure
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Microscopy, Atomic Force
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methods
5.Influence of platelet endothelial cell adhesion molecule-1 on diabetic retinopathy in patients with type 2 diabetes
Jing-dong, ZHANG ; Cheng-ping, CAI ; Ying-feng, WEI ; Nan, HU
Chinese Journal of Experimental Ophthalmology 2012;(12):1127-1130
Background To study diabetic retinopathy (DR) related risk factors is very important in the prevention of DR.Platelet endothelial cell adhesion molecule-1 (PECAM-1) is an important mediator that mediates high blood glucose-induced vascular diseases in diabetic patients.However,its mechanism is still below understood.Objective This clinical study was to investigate the effect of serum level changes of PECAM-1 on DR in type 2 diabetic patients.Methods Fifty-four patients with type 2 diabetes were enrolled from the endocrinology department of the Third People' s Hospital of Nantong City.Fundus examination was performed using the ophthalmoscope and fundus fluorescence angiography (FFA) on all the patients,and these patients were grouped as the non-DR (NDR)group (18 cases),non-proliferative DR(NPDR) group (20 cases) and proliferative DR group (PDR) (16 cases) based on the DR staging criterion of the Chinese Medical Association (1987 version).Eighteen age-and gender-matched normal subjects served as the normal control group.Peripheral blood was collected,and serum PECAM-1 levels were assayed using ELISA.Serum HbA1c levels were detected using the high performance liquid colorimetric(HPLC) method.The correlation of serum PECAM-1 level with serum HbA1c level was analyzed.All results were compared among the groups.Results The serum PECAM-1 levels were (10.907 ± 2.792),(7.024 ±2.377),(5.231 ± 1.816) and (3.817 ± 1.045) μg/L,respectively,in the PDR group,NPDR group,NDR group and normal control group,showing a significant difference among the 4 groups (F =12.630,P =0.02).Serum PECAM-1 content was significantly higher in the PDR group when compared with the NPDR group,NDR group and normal control group (P<0.05).The serum HbA1c levels were (12.596±3.148)%,(9.118±3.356)%,(5.491±1.017)% and (4.992 ± 0.725)% in the PDR group,NPDR group,NDR group and normal control group,respectively,with a significant difference among these 4 groups (F =7.130,P =0.015),and those in the PDR group and NPDR group were significantly elevated in comparison with the NDR group and normal control group (P<0.05).Significantly positive correlations were seen between serum PECAM-1 level and HbA1 c level in the PDR group,NPDR group and NDR group (r=0.799,P<0.01 ;r =0.647,P<0.01 ;r =0.685,P<0.01).Significantly more patients with a disease course of ≥ 10 years were in the NPDR group in comparison with the PDR group (P =0.023).Conclusions Increase of serum PECAM-1 level is closely associated with blood glucose level,and it is an important factor in the pathogenesis and development of DR.These results imply that control of blood glucose is crucial for the prevention of DR in patient with type 2 diabetes.
6.Investigation and analysis of clinical obstetrics and gynecology clinicians' mastery of basic skills and its influencing factors
Chengguang SUN ; Xiaoling GAN ; Shuaibin LIU ; Lina HU ; Xiaojing DONG ; Yiling CAI ; Li LUO
Chinese Journal of Medical Education Research 2015;14(10):1057-1061
Objective To investigate and analyze clinical obstetrics and gynecology clinicians' mastery of basic skills, and then find the way to help clinicians master basic clinical skills. Methods A self-designed questionnaire survey was conducted among 310 clinicians working in 1-3 grade hospital from Chongqing, Sichuan, Guizhou, Yunnan provinces anonymously by spot field investigation and letter-investigations from February 1, 2015 and to March 31, 2015. The data were analyzed by descriptive analysis, using SPSS 19.0 software package for rank sum test analysis. Result Among 310 valid questionnaires, 31.6% (98) clinicians considered having inadequate clinical skills, 26% (81) average and 42.3%(131) expert. All the basic skills obtained had a relationship with the number of operations, working years and professional title. Most of the basic clinical skills were from working in hospitals 85% (270), journal 82% (254), academic conference 58% (174); 94.2% (293) willing to participate in training classes like clinical teaching and training in moulds. Conclusion The gynecology and obstet-rics basic clinical skills have a great relationship with basic skills training. Providing opportunities for the basic clinical skill trainings, academic conferences, in-service education and establishing regional medical source sharing platform can enhance their clinical skills.
7.Curative effect analysis of mild hypothermia in treatment of neonatal hypoxic-ischemic encephalopathy and follow-up study of 36 children aged 18 months
Cheng CAI ; Xiaohui GONG ; Gang QIU ; Dong WEI ; Yong HU ; Chongbing YAN ; Jingjing SUN
Chinese Journal of Applied Clinical Pediatrics 2014;29(24):1858-1861
Objective To explore the efficacy and safety of mild hypothermia (MH) in treating the infants with moderate-to-severe neonatal hypoxic-ischemic encephalopathy(HIE),and to make a follow-up of the nerve motor development of the infants at 18 months old after discharge.Methods Totally 61 neonates with moderate-to-severe HIE in Neonatal Intensive Care Unit (NICU) from Jan.2007 to Dec.2013 were retrospectively analyzed.According to before and after MH therapeutic apparatus was used by NICU of Shanghai Children's Hospital,61 neonates of HIE were divided into 2 groups,the conventional treatment group(25 cases) and MH treatment group(36 cases).The patients in both groups were measured respectively by using the amplitude integrated electroencephalography (aEEG) before MH treatment and at 72 hours after M H treatment,by neonatal behavioral neurological assessment(NBNA) on the 28th day after birth,and by adopting Bayley Scales of Infant Development at 18 months old.The adverse reactions,serious disability cases and deaths of MH treatment were recorded.Results Compared with the conventional treatment group,aEEG recording before treatment showed no statistically significant differences in MH treatment group [maximum voltage:(22.4 ±3.1) μV vs(18.6 ±2.5) μV,maximum voltage:(8.2 ±2.6)μV vs(6.5 ±1.9) μV,t =1.264,0.852,all P > 0.05].However,aEEG recording at 72 h after treatment showed statistically significant differences in MH treatment group [maximum voltage:(24.1 ± 3.2) μV vs (30.6 ± 2.8) μV,maximum voltage:(9.7 ± 3.4) μV vs (13.3 ± 2.2) μV,t =6.376,4.257,all P < 0.05].Severe disability cases [24.0% (6/25 cases) vs 5.6% (2/36 cases),x2 =4.405,P < 0.05] and deaths [16.0% (4/25 cases) vs 0 (0/36 case),x2 =6.1 64,P < 0.05] in MH treatment group were significantly decreased,and there was significantly difference in NBNA on the 28th day after birth[(35.9 ± 2.1) vs(39.1-± 1.6),t =3.361,P < 0.05],and scales of neurobehavioral evaluation through follow-up of 18 months old [mental development index (MDI):(85.2 ± 10.7) vs (96.5-± 13.1),t =7.839,P < 0.05].Very few neonates had apnea,coagulation dysfunction,arrhythmia and other adverse reactions in MH treatment course.Conclusions MH treating moderate-to-severe HIE is safe and effective.MH is effective in reducing death and major disabilities in neonates with moderate-to-severe HIE and without significant side effects.MH can obviously improve the development of nervous system disorders in 0-18 months infants,and can significantly improve these infants' Bayley developmental scale neurobehavioral scores.
8.Intervention effect of erythromycin on tumor necrosis factor-α and interleukin-8 in hyperoxia-induced lung tissue of premature newborn rats
Yihuan CHEN ; Cheng CAI ; Xiaohui GONG ; Gang QIU ; Yong HU ; Dong WEI ; Wenyan HUANG
Chinese Pediatric Emergency Medicine 2015;22(1):18-21
Objective To explore the effect of macrolide antibiotics(erythromycin) on tumor necrosis factor(TNF)-α and interleukin(IL)-8 in hyperoxia-induced lung tissue of premature newborn rats,and to study the intervention effect of erythromycin on hyperoxia-induced lung injury.Methods One-day old preterm Sprague Dawley rats were randomly divided into four groups by random number table method:air + sodium chloride group,air + erythromycin group,hyperoxia + sodium chloride group,hyperoxia + erythromycin group.Hyperoxia groups were continuously exposed to oxygen (oxygen > 0.85) and air group in room air.After 1,7,14 days of exposure,the preterm rats of four groups were sacrificed,whole lung of these rats were isolated,the lung histological changes were observed by hematoxylin-eosin staining,TNF-α and IL-8 in pulmonary tissue homogenate were detected by ELISA.Results The results showed that:(1) Compared with air + sodium chloride group,TNF-α and IL-8 expression in hyperoxia + sodium chloride group were significantly increased(P < 0.05) after 1,7 days of exposure [1 d:TNF-α:(16.163 ± 0.574) ng/ml vs.(21.923 ±2.066) ng/ml,IL-8:(18.214 ±3.649) ng/ml vs.(23.546 ± 5.240) ng/ml ;7 d:TNF-α:(15.940 ±0.821) ng/ml vs.(19.688 ±0.764) ng/ml,IL-8:(18.541 ± 4.114) ng/ml vs.(24.255 ±4.692) ng/ml],in particular,TNF-α expression appeared to increase earlier,their expression became significantly weak in 14 days (P < 0.05).(2) Compared with hyperoxia + sodium chloride group,TNF-α and IL-8 expression in hyperoxia +erythromycin group became significantly weak after 1,7,14 days of exposure(P <0.05) after the intervention of erythromycin [1 d:TNF-α:(21.923 ± 2.066) ng/ml vs.(18.903 ± 1.851) ng/ml,7 d:IL-8:(24.255 ±4.692) ng/ml vs.(23.508 ±3.543) ng/ml,14 d:TNF-α:(16.443 ±5.466) ng/ml vs.(14.453 ±0.963)ng/ml],but their expression became weaker in 14 days than that in 1,7 days.Conclusion The release of inflammatory mediators TNF-α and IL-8 induced by oxidation outbreak participates in the development of hyperoxia induced lung injury,erythromycin may regulate immune function,inhibits the levels of oxidant-mediated TNF-α and IL-8 induced by oxidation outbreak,and alleviate hyperoxia lung injury in premature rats.
9.Effects of erythromycin on glutathione hormone and γ-glutamyl cysteine synthetase in premature newborn rats’ hyperoxia-induced lung injury
Cheng CAI ; Gang QIU ; Xiaohui GONG ; Dong WEI ; Yong HU ; Huanhu ZHAO
Journal of Clinical Pediatrics 2014;(5):476-479
Objectives To explore the effect of erythromycin on glutathione hormone (GSH) and γ-glutamyl cysteine synthetase (γ-GCS) in premature newborn rats exposed to hyperoxia, to study the intervention effect of erythromycin on hype-roxia-induced lung injury. Methods One-day old preterm SD rats were randomly divided into four groups:control group, eryth-romycin group, hyperoxia group, erythromycin+hyperoxia group. Hyperoxia group and hyperoxia+erythromycin group were continuously exposed to oxygen (oxygen concentration>0.85), control group and erythromycin group were in room air. Via cau-dal vein, the preterm rats was injected with erythromycin in erythromycin group and hyperoxia+erythromycin group, sodium chloride in control group and hyperoxia group daily. After 1,7,14 day(s) of hyperoxia (or air ) exposure, the preterm SD rats of four groups were killed, whole lung of these rats were isolated and histological changes were observed by hematoxylin-eosin (HE) staining, GSH andγ-GCS of pulmonary tissue homogenate were detected by double antibody sandwich enzyme linked im-munosorbent assay. Total lung RNA was extracted andγ-GCS mRNA was detected by reverse transcription polymerase chain re-action. Results The results showed that:After 1 and 7 day(s) of exposure, the expression of GSH、γ-GCS andγ-GCS mRNA in four groups showed significant differences(P<0.05). Among them, GSH expression in erythromycin + hyperoxia group was higher than that in the other three groups in 1,7,14 day(s) of exposure with significant differences (P<0.05);GSH expression in erythromycin+hyperoxia group and hyperoxia group reached the peak after 7 days of exposure. The expression ofγ-GCS andγ-GCS mRNA in erythromycin+hyperoxia group and hyperoxia group were higher than the other two groups after 1and 7 day(s) of exposure, the expression ofγ-GCS mRNA in erythromycin+hyperoxia group were higher than that of hyperoxia group with significant differences (P<0.05). Conclusions The expressions of GSH andγ-GCS in the lung of premature SD rats were abnor-mal by oxidation outbreak. Erythromycin may increase the activity ofγ-GCS, improve the anti-oxidation ability of GSH, and al-leviate hyperoxia mediated lung injury in premature rats.
10.Clinical and genetic analysis of a family with low alkaline phosphatase
Weicheng LU ; Congcong SHI ; Dong CAI ; Xu ZHENG ; Hu HAO ; Xin XIAO
Journal of Clinical Pediatrics 2017;35(9):682-686
Objective To investigate the role of TNSALP gene detection in prenatal diagnosis of HPP. Method The clinical data and the results of complete exon sequencing of TNSALP gene in one neonate with low alkaline phosphatase (HPP) were analyzed retrospectively. Peripheral bloods from his family members were collected. The amniotic fluid cell in fetuses at 17 weeks was tested for candidate gene mutations by Sanger sequencing. Results Mainly manifestations in 6-day-old baby were multiple fractures, limb shortening and bending and dyspnea. He died of respiratory failure 9 days after birth. The serum alkaline phosphatase was decreased and serum calcium was decreased slightly; serum phosphorus, serum 25 hydroxyvitamin-D and parathyroid hormone were normal. X-ray showed that the whole body bone was very poorly mineralized, and the long diaphysis was enlarged with shape of a cup at the end and multiple fractures existed. Gene sequencing revealed a complex heterozygous missense mutation in the TNSALP gene, including the heterozygous missense mutation c.542C>T in exon sixth causing 181st amino acids changed from serine to leucine (p.S181L), and tenth exon heterozygous missense mutation in c.1016G>A causing 339th amino acid changed from glycine to glutamic acid (p.G339E). The parental phenotypes were normal. The c.542C>T mutation is inherited from his father and the c.1016G>A mutation is inherited from his mother. These two mutations were not detected in the fetus. Conclusion TNSALP gene analysis can be applied to the diagnosis and prenatal diagnosis of HPP.