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.Effect of MKK34 on airway β-catenin in a HDM-induced mouse asthma
Yahui HU ; Hangming DONG ; Laiyu LIU ; Chaowen HUANG ; Lishan LUO ; Xuan WAN ; Haijin ZHAO ; Shaoxi CAI
The Journal of Practical Medicine 2017;33(4):543-547
Objective To explore the role of MKK34 (a peptide spanning a C-terminal α-helical region in TSLP) on airway inflammation and β-catenin of airway epithelium in a HDM-induced mouse asthma.Methods 32 male BALB/c mice were randomly divided into control,MKK34,asthma and MKK34 + HDM groups.The mice in the asthma group were exposed to HDM for five consecutive days and the MKK34 + HDM group was pretreated with MKK34 1 h prior to the HDM intranasally treated.After 8 weeks' treatment,animal lung function test and pathological staining were performed to evaluate the asthma situation,IL-4,IFN-γin bronchoalveolar lavage fluid and IgE in the serum were detected,immunohistochemistry and western blot were used to assess β-catenin and p-ERK,t-ERK levels.Results Airway reactivity,IL-4 and IgE in the asthma group were significantly higher than that in the control group.Treatment with MKK34 significantly decreased airway hyperresponsiveness,IL-4 and IgE.HE staining demonstrated the chronic bronchitic inflammation in the lungs of asthma group.β-catenin in the control group was distributed evenly at the cytomembrane of epithelial cells.In the asthma group,β-catenin was disordered in epithelial cells and its expression was decreased.Treatment with MKK34 ameliorated the damage of β-catenin and chronic bronchitic inflammation.The protein levels of p-ERK1/2 increased obviously in the asthma group.The pretreated group significantly decreased the expression of p-ERK1/2.Conclusions MKK34 can ameliorate the airway inflammation and the destruction of β-catenin of airway epithelium in a HDM-induced mouse asthma.The ERK pathway may play a role in this process.
6.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.
7.Clinical research of free vascularized fibular grafting for osteonecrosis of femoral head
Shuai DONG ; Xiyu CAI ; Zhengwei ZHU ; Jie LI ; Shuichang ZHENG ; Dajiao HU ; Yi LI ; Qingju WU
Chinese Journal of Microsurgery 2017;40(3):237-240
Objective To evaluate the clinical results of free vascularized fibular grafting (FVFG) for the treatment of osteonecrosis of the femoral head (ONFH).Methods From July,2009 to January,2013,85 cases (120 hips) of ONFH were treated with free vascularized fibular grafting.These cases included 61 males (87 hips) and 24 females (33 hips) with an average age of 36.5 years (22-51 years);7 hips (Ⅰ A 2 hips,Ⅰ B 3 hips,Ⅰ C 2 hips) were at stage Ⅰ,98 hips (Ⅱ A 24 hips,ⅡB 39 hips,ⅡC 35 hips) at stage Ⅱ and 15 hips (ⅢA 9 hips,ⅢB 4 hips,Ⅱ C 2 hips) at stage]Ⅲ according to the classification system of Association Research Circulation Osseous (ARCO).The mean preoperative Harris hip score was (60.21±6.85) points (42-71 points),The follow-up items included the X-ray examination,the Harris scores of the hip,and the evaluation of the complications.Results Eighty-three cases (117 hips) were followed up.The average duration of follow-up was 25 months (range from 12 months to 42 months).The mean postoperative Harris hip score was increased to (81.26±5.84) points (67-91 points) by the end of the follow-up,compared with the preoperation,the score improved significantly,the difference was statistically significant (P<0.05).Comparing with postoperative X-ray,101 hips (86.3%) were improved,12 hips (10.3%) had no significant changes and deterioration occurred in 4 hips (3.4%).Conclusion The free vascularized fibular grafting is an effective method for treating osteonecrosis of and preventing the collapse of the femoral head.
9.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.
10.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.