1.A mice model for continuous, dynamic and direct observation of skin wound infection
Youzhen SHI ; Ying WANG ; Chuan GU ; Zhigang JIA ; Xiujun FU ; Yong FANG ; Weirong YU ; Min YAO
Chinese Journal of Trauma 2012;28(9):854-858
Objective To establish a stable animal model for sequentially dynamic and direct monitoring of the skin wound infection. Methods The mice with full-thickness skin incisions were replicated. After immediate subcutaneous suture,the mice were randomly divided into four groups,ie,Group A was inoculated with 50 μl sterile PBS solution),Groups B,C and D were inoculated with 50 μl suspension containing 1 × 106,1 × 108 and 1 × 1010 colony forming unit (CFU)/ml bioluminescent methicillin-resistant staphylococcus aureus (MRSA) respectively.Then,the diet behavior of each group was observed and the mean weight and mortality of each group were also recorded at different time points.The bioluminescent intensity of fluoresce in the wounds was recorded at different time points by using the charge-coupled device (CCD) based imaging system.Local wound tissues were incised at 24 hours after inoculation for HE staining so as to observe wound inflammatory reaction in each group.Wound healing time of each group was also recorded. Results ( 1 ) Average weight:Groups A and B showed unobvious changes in weight; Group C lightened until day 3 after inoculation and then recovered gradually to the preinoculation level at day 14; Group D lightened gradually until death.(2)Mortality:Groups A and B had no death; Group C had 10% deaths at day 14; Group D had 100% deaths.(3) Bioluminescent intensity of wounds:Groups A and B showed a gradual weakened luminescence since the day of inoculation and had almost complete disappearance at days 5 and 7 respectively; there was no sign of obvious increase or decrease in Group C from the day of inoculation till day 14 ; Group D had a gradual increase since the day of inoculation and the luminous area expanded until the death.(4) HE staining at 24 hours after inoculation:all the four groups showed inflammatory cell infiltration,especially in Groups C and D.(5) Wound healing time:wound healed at days 5 and 7 after inoculation in Groups A and B; the wounds showed no healing even at day 14 in the Group C,but the wounds length and area did not show obvious enlargement or diminishment ; the wounds extended gradually until the death in the Group D,since the day of inoculation. Conclusions The inoculation of 50 μl suspension with 1 × 108 CFU/ml bioluminescent MRSA to full-thickness skin incision rats allows direct,real-time dynamic and continuous detection of the occurrence and development of the wound infections.The infection model is easy to make and has stability and high repeatability.
2.Repair of corneal perforation with sutureless photochemical tissue bonding
Ying WANG ; Yong FANG ; Weirong YU ; Youzhen SHI ; Chuan GU ; Min YAO
Chinese Journal of Trauma 2012;28(1):79-82
ObjectiveTo develop a new sutureless technique (photochemical tissue bonding,PTB ) for repair of corneal perforation. Methods A total of 60 rabbits were used for creating corneal perforation models.The corneal perforation on the left eye was repaired by sutures and the injury on the right eye was fixed with the use of amniotic membrane with PTB.The outcomes of the two mentioned repair methods were compared by observing the leakage of aqueous and the morphology of the anterior chamber at different instants,measuring the intraocular pressure (IOP) and observing the formation of neo-vessels and scars of cornea in the use of histological analysis. Results There was no leakage of aqueous and no difference for morphology evaluation in both treatments.PTB could adhere AM on the cornea to restore the corneal perforation.The peak IOP in the PTB treatment group at days 0,7 and 14 postoperative [ (531.2 ±49.5) mm Hg,(542.6 ±74.8) mm Hg and (603.9 ±69.1) mm Hg,respectively] was significantly higher than that in the suture group at the same instants [ (41.3 ±12.7) mm Hg,(142.6 ±25.4) mm Hg and (333.3 ± 66.7) mm Hg,respectively] (P <0.O1 ).Compared with suture repair,the treatment with PTB resulted in a better outcome of wound healing with less neo-vessels and less scars of cornea. Conclusion PTB treatment for repair of corneal perforation is superior to suture repair.
3.Cloning of human uracil N-glycosylase and its detection in cancer tissues by quantitative RT-PCR.
Hong-Bo BAO ; Chuan-Bao ZHANG ; Jin-Fang WANG ; Chuan-Nong ZHOU ; Fang LIU ; Xiao-Hang ZHAO ; Shi-Jun QIAN
Chinese Journal of Biotechnology 2003;19(5):561-565
The uracil in DNA comes from either the misincorporation of dUTP in place of dTTP or deamination of cytosine. In the latter case, it can result in a GC to AT transition mutation if the uracil is not removed before DNA replication. Base excision repair (BER) is a major pathway for removing DNA lesions arising from endogenous processes as well as those induced by exposure to exogenous chemicals or irradiation. BER is initiated by DNA glycosylases that excise aberrant bases from DNA by cleavage of the N-glycosidic bond linking to the base of its deoxyribose sugar. Uracil N-glycosylase (UNG) is the enzyme responsible for the first step in the BER pathway that specifically removes uracil from DNA. The UNG gene undergoes both temporal and spatial regulation mainly at the level of transcription. Normally cancer cells undergo over-proliferation and up-regulate their UNG during tumorigenesis. In this study we examine the correlation between UNG level and carcinogenesis, and explore the possibility of using UNG as a marker for cancer diagnosis. Human UNG gene was amplified from the total RNA of the human choriocarcinoma cell line, JEG-3, by RT-PCR. After purification, the 942bp full-length UNG cDNA coding sequence was digested with EcoR I and Sal I, and cloned into the digested pET-21 to construct a recombinant vector, pUNG. The UNG protein was expressed under the control of T7 promoter in E. coli BL21 (DE3) cells induced with IPTG. After ultrasonic treatment, the cell lysate and precipitate were analyzed by SDS-PAGE and a 39kD band was detected. The plasmid was serially diluted at appropriate concentrations and employed as standards in the subsequent quantification. Total RNAs were extracted from 18 pairs of clinical samples, each pair contains a sample of esophageal squamous cell carcinoma (ESCC) tissue and its surrounding normal esophageal epithelia. The copy numbers of UNG mRNA in these RNA samples were determined by real-time quantitative RT-PCR using a Lightcycler (Roche). UNG was present in 13 cases of ESCC (13/18, n = 18) but absent in all of the normal tissues. The results indicated that there was a correlation between high level of UNG expression and the carcinogenesis of ESCC.
Carcinoma, Squamous Cell
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genetics
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metabolism
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Cell Line, Tumor
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Cloning, Molecular
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Electrophoresis, Polyacrylamide Gel
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Esophageal Neoplasms
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genetics
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metabolism
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Humans
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In Vitro Techniques
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Reverse Transcriptase Polymerase Chain Reaction
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Uracil-DNA Glycosidase
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genetics
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metabolism
4.Influence of Wenxin particle on cardiac electrophysiology in depressed rats after myocardial infarction
Jinjun LIANG ; Shaobo SHI ; Jingjing SHEN ; Fang WANG ; Yingying SHEN ; Xiaoran YUAN ; Chuan QU ; He HUANG ; Bo YANG ; Congxin HUANG
Chinese Journal of cardiovascular Rehabilitation Medicine 2012;21(6):649-655
Objective: To investigate influence of Wenxin particle on cardiac electrophysiology in rats complicated with depression after myocardial infarction (MI). Methods: A total of 50 SD rats were randomly and equally divided into normal group, MI group, depression group, MI complicated with depression group (model group) and Wenxin particle group (Wenxin particle were given to model rats by gavage, 2 g/d, 28d). Model(MI complicated with depression)rats were made by acute ligation of left coronary artery and supply chronic unpredictable mild stress in order. The influences of Wenxin particle on cardiac electrophysiological indexes, such as monophasic action potential (MAP90), left ventricular effective refractory period (ERP) and ventricular fibrillation threshold (VFT) were evaluated in Wenxin particle group. Results: (1) Compared with normal group, there were significant decrease in behavior scores (P<0.05) in model group, after four-week treatment with Wenxin particle, their behavior scores significantly increased (P<0.01); (2) Compared with normal group, there were significant increase in MAPD90 and ERP, and significant decrease in VFT in model group (P<0.05); compared with model group, there were significant decrease in MAPD90 [(89.33±7.12) ms vs. (72.29±8.37) ms] and ERP [(84.00±6.57) ms vs. (68.00±7.43) ms], and significant increase in VFT [(7±3.11)V vs. (29±5.60)V] in Wenxin particle group, P<0.05. Conclusion: Wenxin particle can improve cardiac electrical remodeling in rats complicated with depression after myocardial infarction, including decrease monophasic action potential duration and effective refractory period, and raise ventricular fibrillation threshold.
5.Establishment and analysis of chronic periodontitis and atherosclerosis model in Wistar rat.
Liang-Jun ZHONG ; Jun XU ; Yuan-Ming ZHANG ; Jia NI ; Xiao-Huan ZHOU ; Fang-Chuan SHI
Chinese Journal of Stomatology 2009;44(8):464-468
OBJECTIVETo investigate the possible correlation between atherosclerosis and chronic periodontitis by establishing an animal model of chronic periodontitis and atherosclerosis in Wistar rat.
METHODSSixty male Wistar rats were divided into four groups: A (control group), B (chronic periodontitis group), C (atherosclerosis group), D (chronic periodontitis accompany with atherosclerosis group). Every group was accepted the corresponding treatment. Animals were sacrificed after 12 weeks. The periodontal index, levels of serum total cholesterol (TC) and low-density lipoprotein (LDL), the concentration of TNF-alpha and matrix metalloproteinase (MMP-3) were examined. The severity of chronic periodontitis and atherosclerosis was quantified by histopathology. The date were statistically analyzed.
RESULTSThrough detection of periodontal tissue of experimental teeth, serum and histopathology, animal models were successful. Histopathologic observation revealed:obvious inflammation of periodontal tissue was observed in group B and D. Attachment loss level in group B [(137.86 +/- 28.39) microm] and D [(162.36 +/- 22.69) microm] was higher than that in group A [(4.26 +/- 1.07) microm] and C [(68.07 +/- 18.25) microm] (P < 0.05), and that in group C was higher than group A (P < 0.05). Atherosclerotic lesions of abdominal aorta were formed in group C and D. The level of TC, LDL in group C and D was higher than that in group A and B (P < 0.05), and that in group D was higher than group C (P < 0.05). Animals in group B and D showed higher level of TNF-alpha, MMP-3 in serum than that in group A and C (P < 0.05). There was no correlation between the level of MMP-3 and TC (P = 0.971) or LDL (P = 0.604).
CONCLUSIONSChronic periodontitis may be a risk factor and contribute to the pathogenesis of atherosclerosis. MMP-3 may be an independent risk factor of atherosclerosis exclude TC and LDL.
Animals ; Aorta, Abdominal ; Aortic Diseases ; etiology ; pathology ; Atherosclerosis ; etiology ; pathology ; Cholesterol ; blood ; Chronic Periodontitis ; complications ; pathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; Lipoproteins, LDL ; blood ; Male ; Matrix Metalloproteinase 3 ; blood ; Periodontal Index ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood
6.Expression of tumor necrosis factor in placenta tissue of pregnant rats with chronic peridontitis.
Jing LIU ; Liang-jun ZHONG ; Fang-chuan SHI ; Lei ZHANG ; Qiong ZHANG ; Da-lei SUN
Chinese Journal of Stomatology 2011;46(5):276-279
OBJECTIVETo examine the expression of tumor necrosis factor in placenta of pregnant rats with chronic periodontitis.
METHODSTwenty Wistar female rats were randomly divided into two groups, control (n = 8) and experimental group (n = 12). The periodontitis model was established in the experimental group. The females and males in the two groups got together four weeks later. Nineteen days after pregnancy all rats were executed and placenta collected. The delivery time and neonatal birth weight were recorded and the pathological changes of periodontal tissue observed. Tumor necrosis factor (TNF) expression was examined in placenta by real-time quantitative polymerase chain reaction analysis.
RESULTSThe animal model of chronic periodontitis was successfully established. Experimental group delivered 30 offspring and the control group 56 offspring. The average number of pups born alive per litter in experimental group (4.1 ± 2.2) was significantly lower than that in control group (9.2 ± 2.2), P < 0.05. The birth weight of pups in experimental group [(5.01 ± 0.43) g] was significantly lower than that in the control group [(5.79 ± 0.53) g], P < 0.05. The relative quantitative expression of TNF was (1.807 ± 0.265) in experimental group the and (1.003 ± 0.021) in the control group (P = 0.001).
CONCLUSIONSChronic periodontitis may be related to preterm low birth weight.
Aggregatibacter actinomycetemcomitans ; Animals ; Animals, Newborn ; Birth Weight ; Chronic Periodontitis ; metabolism ; microbiology ; Disease Models, Animal ; Female ; Fusobacterium nucleatum ; Placenta ; metabolism ; Porphyromonas gingivalis ; Pregnancy ; Pregnancy Complications, Infectious ; metabolism ; microbiology ; Prevotella intermedia ; Random Allocation ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; metabolism
8.Research progress of inflammatory immunity and regulation mechanism of interleukin-38
Yu-Sen CHAI ; Chuan-Jiang WANG ; Shi-Hui LIN ; Fang XU
Chinese Journal of Immunology 2019;35(2):230-235
Interleukin ( IL) -38 is the most recently discovered cytokine, which is the tenth member of IL-1 cytokine family. IL-38 shares a high homology with IL-1 receptor antagonist ( IL-1 Ra) and IL-36 Ra. IL-36 R is the specific receptor of IL-38 which is the partial receptor antagonist of IL-36. Now, research has shown that IL-38 not only had relationship with intracellular infection and inflammation, but also with many clinical diseases such as autoimmune diseases, osteoarthritis, tumor and cardiopulmonary diseases. However, the molecular immune regulation mechanism of IL-38 is complicated, involving specific receptor recruitment, apoptosis mediated phagocyte regulation, regulation of Th17 cell differentiation and many other processes. It also provides several new clues to the treatment of diseases. IL-38 is a new cytokine research hot spot with extensive application prospects and need deep exploration.
9.The impact of childhood sexual abuse on the development of AIDS related high risk behaviors and psychological appearances among men who have sex with men
Zeng-Zhao YU ; Bei-Chuan ZHANG ; Xiu-Fang LI ; Ning WANG ; Tong-Xin SHI ; Quan-Sheng CHU
Chinese Journal of Epidemiology 2009;30(1):14-17
Objective To study the AIDS related high risk behaviors and psychological appearances among men who have sex with men (MSM) who ever experienced childhood sexual abuse (CSA).Methods Target sampling for a cross-sectional study was developed and valid anonymous questionnaires were adopted to compare the differences of high risk behaviors related to AIDS and psychological appearances betwen those with or without CSA experiences among 2147 MSM from nine cities.Results Compared to corresponding ones without CSA experience,CSA group had a significant larger numbers in the following events: total sexual partners,anal sex episodes with same sex,female sexual partners and anal sex in the previous six months,with the figures of median as 20.0,10.0,3.0,3.0 respectively.In the previous year,30.8% of them had ever participated in 'group sex',19.2% ever exchanged money for sex,36.7% bled while having sexual intercourse,37.3% had sex with male partners away from his own region.All the above said figures were higher than non-CSA group,with significant differences.It also appeared that CSA experience had an impact on significant lower rate of condom use (67.3%) in the last anal sex.Those with CSA experience had more psychological problems which appeared as: 75.6% considered they would suffer from serious discrimination if their sexual orientation ever disclosed,34.7% had a strong intention of suicide and 24.3% ever having had suicidal attempts.The differences of the two groups showed statistical significance.Condusion CSA experience not only increased the number of AIDS related high risk behaviors in adulthood,but also had negative impact on their psychological appearances.It is of urgent need to carry out psychological intervention approaches to target on MSM with CSA experiences while childhood sexual education and rights assurance towards juvenile population should also not be neglected.
10.Neurocognition and Function of Social Life Investigation among HIV/AIDS Subjects Infected by Plasma Donation
Chuan SHI ; Xin YU ; Ying LIANG ; Yu-Fang CHEN ; Zun-You WU ; Hua JIN ; Ofilio VIGIL ; Lucette A.CYSIQUE ; Donald R.FRANKLIN Jr ; Igor GRANT ; Robert K.HEATON
Chinese Mental Health Journal 2009;23(11):800-805
Objective:To investigate the neurocognition and function of social life among human immunodeficiency virus (HIV) positive subjects infected by plasma donation.Methods:Totally 203 HIV positive subjects infected by plasma donation were recruited.Neuropsychological (NP) battery tests including executive function,verbal fluency,learning,memory,fine motor skill,speed of information processing and working memory domains were performanced among these subjects,as well as the Activity of Daily Living Scale (ADLs).And 198 HIV negative plasma donors were matched in gender,age and years of schooling.The cut-off of Global Deficit Score (GDS) was 0.5.Results:The HIV positive subjects performanced worse in global cognitive function [(45.7±5.9 vs.(49.4±6.0),P<0.01]and 7 cognitive domains than those of HIV negative subjects.HIV and HCV infection interacted on aspect of fine motor skill (F=5.28,P<0.05).The impairment rate of the subjects in asymptomatic stage and AIDS were 29.2% and 43.0% respectively.The HIV positive subjects showed more decline of ADLs scores than that of HIV negative subjects [(0.49±1.32) vs.(0.14±0.75),P<0.01].Working months,individual and family's income of HIV positive subjects were also less than that of HIV negative subjects(P<0.01).The lowest CD4 count in NP-impaired group was lower than that in non-impaired group [(214.3±144.0) vs.(274.8±161.1),P=0.01].Subjects with NP impairment reported more decline of ADLs scores [(0.75±1.58) vs.(0.34±1.13),P<0.01] than those without NP impairment.The global cognitive function scores were correlated with decline of ADLs scores (r=-0.22,P<0.01).Conclusion:HIV positive subjects are impairment on aspect of 7 cognitive domains,and co-infection of HCV can lead to more fine motor skill impairment.The more progression of disease,the higher NP impairment ratio.HIV infected subjects show decline of ADL,poorer occupational function and economical status.The lowest CD4 count may be a predictor to NP impairment.NP impairment is a factor which influenced the activity of daily living and the income of individual and family.