[Show abstract][Hide abstract] ABSTRACT: To investigate the prevalence and diagnosis rate of intra-abdominal hypertension (IAH) in a mixed-population intensive care unit (ICU), and to investigate the knowledge of ICU staff regarding the guidelines published by the World Society of Abdominal Compartment Syndrome (WSACS) in 2013.
Preview · Article · Dec 2015 · Chinese Journal of Traumatology (English Edition)
[Show abstract][Hide abstract] ABSTRACT: To compare the predictive value of anatomic scoring system, physiological scoring system, and the combination of two systems in death prediction of patients with severe trauma in intensive care unit ( ICU ).
A retrospective analysis of patients with severe trauma admitted to department of critical care medicine of Daping Hospital, the Third Military Medical University, and Zunyi Medical University from January 2011 to December 2014 was conducted. The patients meeting the following criteria were enrolled: over 16 years old, admitted to hospital shorter than 24 hours after trauma, length of ICU stay≥48 hours, and injury severity score ( ISS ) ≥16. Patients were divided into two groups: survivors and non-survivors. The data of anatomic scoring system, including ISS and new injury severity score ( NISS ), and physiological scoring system, including acute physiology and chronic health evaluation II ( APACHEII) score were collected. The predictive power for death of the scoring system alone or combination in patients with severe trauma was evaluated.
A total of 614 patients with severe trauma were enrolled, and there were 153 deaths with a mortality rate of 24.9%. ISS, NISS, APACHEII, ISS + APACHEII, NISS + APACHEII of non-survivors were significantly higher than those of survivors ( ISS: 29.15±7.75 vs. 24.31±6.50, NISS: 41.96±12.01 vs. 29.64±8.19, APACHEII: 23.71±6.58 vs. 17.02±5.49, ISS + APACHEII: 52.86±10.00 vs. 41.33±8.70, NISS + APACHEII: 65.67±13.46 vs. 46.66±10.43, all P < 0.01 ). The area under receiver operating characteristic curve ( AUC ) of ISS, NISS, APACHEII, ISS + APACHEII, NISS + APACHEII was 0.687, 0.792, 0.782, 0.809, and 0.860, respectively. Both of ISS + APACHEII and NISS + APACHEII had higher AUC than that of ISS, NISS or APACHEII alone; and the AUC of NISS + APACHEII was significantly larger than that of ISS + APACHEII ( all P < 0.05 ). NISS + APACHEII showed the largest AUC in death prediction of severe trauma patients. The cut-off value, sensitivity, specificity, positive predict value ( +PV ), negative predict value ( -PV ), positive likelihood ratio ( +LR ), negative likelihood ratio ( -LR ), and Youden index of NISS + APACHEII, which had the greatest AUC, were 56, 75.2%, 82.0%, 58.1%, 90.9%, 4.17, 0.30, and 0.572, respectively.
The combination of anatomic scoring system and physiological scoring system is better than single scoring system for death prediction in patients with severe trauma in ICU, and it may be considered to be a new method for early identification of death risk in patients with severe trauma.
No preview · Article · Apr 2015 · Zhonghua wei zhong bing ji jiu yi xue
[Show abstract][Hide abstract] ABSTRACT: Table S1. Primers of the four variants of the RAGE gene and their PCR conditions. The PCR primers, sequencing primers and the annealing temperatures of the four variants of the RAGE gene were shown in Table S1.
[Show abstract][Hide abstract] ABSTRACT: The methods of plasmid construction. The possible effect of -429T/C on the promoter activity was investigated using a reporter gene assay system. Two plasmids which contained the -429T promoter and -429C promoter were constructed.
[Show abstract][Hide abstract] ABSTRACT: Introduction
The receptor for advanced glycation end products (RAGE) has been considered as one of the major pattern recognition receptors and plays an important role in the development of sepsis and multiple organ dysfunction in critical illnesses. Although genetic variants of the RAGE gene have been shown to be well associated with susceptibility to some inflammatory diseases, little is known about their clinical relevance in the development of sepsis in critical ill patients.
Four genetic variants were selected from the entire RAGE gene and genotyped using pyrosequencing and polymerase chain reaction-length polymorphism methods. Association studies were performed in two independent Chinese Han populations.
Among the four genetic variants, only the rs1800625 polymorphism was significantly associated with sepsis morbidity rate and multiple organ dysfunction (MOD) scores in patients with major trauma both in Chongqing (n = 496) and Zhejiang (n = 232) districts, respectively. Results from ex vivo responsiveness of peripheral blood leukocytes indicated that the rs1800625 polymorphism was well associated with decreased production of TNFα. In addition, the rs1800625 polymorphism could significantly inhibit the promoter activities of the RAGE gene.
The rs1800625 polymorphism is a functional variant, which might be used as a relevant risk estimate for the development of sepsis and multiple organ dysfunction syndrome in patients with major trauma.
[Show abstract][Hide abstract] ABSTRACT: Myeloid differentiation 2 (MD-2) plays a critical role in orchestrating the innate immune response and the development of sepsis and subsequent organ dysfunction after trauma. The objectives of this prospective study were to identify haplotype tag single-nucleotide polymorphisms (htSNPs) within the entire MD-2 gene and to investigate their clinical relevance in patients with major trauma. A total of 726 patients with major trauma were prospectively recruited and composed of two different geographic populations (Chongqing in southwestern China and Zhejiang in eastern China). The htSNPs of the MD-2 gene were determined using HapMap database and linkage disequilibrium analysis. The htSNPs were genotyped using pyrosequencing method. The whole peripheral blood samples obtained immediately after admission were stimulated with bacterial lipopolysaccharide and then determined for production of tumor necrosis factor α. Sepsis morbidity rate and multiple organ dysfunction (MOD) scores were accessed. Three SNPs (rs7843858, rs11465996, and rs2114169) were identified as htSNPs for the MD-2 gene. All of them were shown to be high-frequent SNPs in this study cohort. However, only the rs11465996 polymorphism was shown to be significantly associated with higher sepsis morbidity rate and MOD scores in patients with major trauma in both Chongqing and Zhejiang districts. In addition, the rs11465996 polymorphism was significantly associated with tumor necrosis factor α production by peripheral blood leukocytes in response to bacterial lipoprotein stimulation. Among the three htSNPs of the entire MD-2 gene, only the rs11465996 might be used as relevant risk estimate for the development of sepsis and MOD syndrome in patients with major trauma.
No preview · Article · Jan 2012 · Shock (Augusta, Ga.)
[Show abstract][Hide abstract] ABSTRACT: The nucleotide-binding oligomerization domain-like receptor (NLR) family has been recognized as comprising intracellular pattern recognition receptors in which NLRP3 (NLR family, pyrin domain containing 3) plays an important role in the initiation of host immune inflammatory responses. The genetic variants have been recognized to be critical determinants of interindividual differences in both inflammatory responses and clinical outcomes in critical illness. However, little is known about the clinical relevance of NLRP3 gene polymorphisms in critical illness.
A total of 718 patients with major blunt trauma were included in this study. Six tag SNPs (tSNPs) were selected from the entire NLRP3 gene through construction of haplotype bins, and they were genotyped using a pyrosequencing method. They were analyzed in relation to sepsis morbidity rate, multiple organ dysfunction (MOD) scores and IL-1β production. Moreover, the functionality of the rs2027432 polymorphism was assessed by the observation of its effect on transcriptional activities.
Among the six tSNPs genotyped in this study, two of them (rs2027432 and rs12048215) were significantly associated with sepsis morbidity rate and MOD scores. A significant association was also observed between these two polymorphisms and IL-1β production by peripheral leukocytes in response to ex vivo lipopolysaccharide stimulation. However, no combined effects were found between these two polymorphisms. In addition, the rs2027432 polymorphism could significantly enhance the promoter activities of the NLRP3 gene.
rs2027432 and rs12048215 polymorphisms might be used as relevant risk estimates for the development of sepsis and MOD syndrome in patients with major trauma, in which rs2027432 might be a functional SNP.
Full-text · Article · Nov 2011 · Critical care (London, England)
[Show abstract][Hide abstract] ABSTRACT: Tumor necrosis factor (TNF) is a proinflammatory cytokine that plays a major role in the sepsis and multiple organ dysfunction secondary to major trauma. The purpose of this article was to research the clinical relevance of the TNF gene polymorphism in patients with major trauma.
Three hundred six patients with major trauma were prospectively recruited. The TNF gene polymorphisms were genotyped using restriction fragment length polymorphism analysis. Plasma TNF-α levels were determined with enzyme-linked immunosorbent assay. Sepsis morbidity rate and multiple organ dysfunction scores were accessed.
The TNF-α/-308 polymorphism was shown to be well associated with increased capacity of peripheral leukocytes to produce TNF-α in response to ex vivo lipopolysaccharide stimulation in trauma patients at admission. Results from association study indicated that trauma patients carrying the TNF-α/-308/A allele were more likely complicated with sepsis.
The TNF-α/-308 polymorphism might be used as a biomarker for the assessment of outcome of trauma patients, but the TNF-β/252 gene polymorphism might not influence the development of complications in patients with major trauma.
No preview · Article · Apr 2011 · The Journal of trauma
[Show abstract][Hide abstract] ABSTRACT: To investigate the surgical treatment for patients with multiple injuries in ICU.
Clinical data of 163 multiple injury patients admitted to ICU of our hospital from January 2006 to January 2009 were retrospectively studied, including 118 males and 45 females, with the mean age of 36.2 years (range, 5-67 years). The injury regions included head and neck (29 cases), face (32 cases), chest (89 cases), abdomen (77 cases), pelvis and limbs (91 cases) and body surface (83 cases). There were 57 cases combined with shock. ISS values varied from 10 to 54, 18.42 on average. Patients received surgical treatments in ICU within respectively 24 hours (10 cases), 24-48 hours (8 cases), 3-7 days (7 cases) and 8-14 days (23 cases).
For the 163 patients, the duration of ICU stay ranged from 2 to 29 days, with the average value of 7.56 days. Among them, 143 were cured (87.73%), 11 died in the hospital (6.75%) due to severe hemorrhagic shock (6 cases), craniocerebral injury (3 cases) and multiple organ failure (2 cases), and 9 died after voluntarily discharging from hospital (5.52%). The total mortality rate was 12.27%.
The damage control principle should be followed when multiple injury patients are resuscitated in ICU. Surgical treatment strategies include actively controlling hemorrhage, treating the previously missed injuries and related wounds or surgical complications and performing planned staging operations.
Full-text · Article · Feb 2011 · Chinese Journal of Traumatology (English Edition)
[Show abstract][Hide abstract] ABSTRACT: Toll-like receptor 2 (TLR2) signaling plays a critical role in orchestrating the innate immune response and the development of sepsis and subsequent organ dysfunction after trauma. The objectives of this prospective study were to identify haplotype tag single-nucleotide polymorphisms (htSNPs) within the entire TLR2 gene and to investigate their clinical relevance in patients with major trauma. A total of 410 patients with major trauma were prospectively recruited. The htSNPs of the TLR2 gene was determined using HapMap database and linkage disequilibrium analysis. The htSNPs were genotyped using polymerase chain reaction-restriction fragment length polymorphism method. The whole peripheral blood samples obtained immediately after admission were stimulated with bacterial lipoprotein and then determined for production of tumor necrosis factor-α, interleukin 8, and interleukin 10. Sepsis morbidity rate and multiple organ dysfunction (MOD) scores were accessed. Three SNPs (rs1898830, rs3804099, and rs7656411) were identified as htSNPs for the TLR2 gene. All of them were shown to be high-frequency SNPs in this study cohort. Two of them (rs1898830 and rs3804099) and the haplotype ATT were significantly associated with cytokine production by peripheral blood leukocytes in response to bacterial lipoprotein stimulation. Only rs3804099, however, was significantly associated with higher sepsis morbidity rate and MOD scores in patients with major trauma. In addition, the patients with the haplotype ATT had lower sepsis morbidity rate than those without the haplotype ATT. Therefore, three SNPs might act as htSNPs for the entire TLR2 gene in the Chinese population. The rs3804099 and the haplotype ATT might be used as relevant risk estimates for the development of sepsis and MOD in patients with major trauma.
No preview · Article · Jan 2011 · Shock (Augusta, Ga.)
[Show abstract][Hide abstract] ABSTRACT: To investigate the possibility that the level of Tie2 mRNA in peripheral blood could reflect the severity of sepsis.
Trauma patients in intensive care unit (ICU) were recruited, and they were divided into sepsis group (n=13) and non-sepsis group (n=19). The severity of disease was evaluated with acute physiology and chronic health evaluation II (APACHEII) score on the day of ICU admission. Blood of patients was sampled on day 1, 3, 7 after ICU admission to determination of the white blood cell (WBC) count, contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN) and creatinine (Cr), mRNA levels of Tie2 in blood measured by quantitative real-time polymerase chain reaction (PCR), and the contents of plasma von Willebrand factor (vWF) with enzyme linked immunosorbent assay (ELISA).
No significant difference in contents of ALT, AST and plasma vWF was found between sepsis group and non-sepsis group [ALT (U/L): 53.30 (199.58) vs. 80.65 (202.62), AST (U/L): 316.53 (49.90) vs. 66.10 (285.03), vWF: (272.47+/-114.61)% vs. (246.66+/-128.77)%, all P>0.05]. The number of WBC [x10(9)/L, 18.26 (21.82) vs. 10.11 (4.72)], the contents of BUN [mmol/L, 20.70 (11.20) vs. 7.70 (5.45)] and Cr [micromol/L: 252.00 (364.55) vs. 68.00(23.20)], and the circulating mRNA levels of Tie2 (1.86+/-0.67 vs. 0.91+/-0.42) in sepsis group were higher than those in the non-sepsis group (all P<0.01). The Tie2 mRNA level in peripheral blood of each patient was positively correlated with APACHEII score (r=0.532, P<0.01). The linear regression equation was Y=12.66+4.922 X (R2=0.283). Besides, there was a significant correlation between the amount of Tie2 mRNA and plasma levels of vWF (r=0.334, P<0.05). The linear regression equation was Y=180.932+57.93 X (R2=0.112).
The level of Tie2 mRNA in peripheral blood could reflect the damage of endothelial cell and severity of sepsis.
No preview · Article · Jun 2010 · Zhongguo wei zhong bing ji jiu yi xue = Chinese critical care medicine = Zhongguo weizhongbing jijiuyixue
[Show abstract][Hide abstract] ABSTRACT: The goal of this study was to establish a convenient and effective approach to anti-inflammation treatment by rebalancing the sympathetic-vagal system via vagal nerve stimulation (VNS). We established an endotoxemia model in Sprague-Dawley rats using lipopolysaccharide (LPS) injection. Electrical discharges in the vagal system, including the nucleus tractus solitarii (NTS) and afferent and efferent cervical vagal nerves, were detected. The condition of sympathetic-vagal balance, presented as heart rate variability (HRV) and hepatic norepinephrine/acetylcholine (NE/ACh), was measured following endotoxemia with and without VNS. Discharges in afferent and efferent vagal nerves increased significantly following LPS injection compared with the basis level and corresponding time points in the control group. Discharges in the NTS also increased significantly following LPS injection. The HRV components, including normalized high frequency (HFnm), normalized low frequency (LFnm), LF/HF, and very low frequency (VLF), increased significantly following LPS injection. HFnm values in the LPS + VNS group increased significantly compared with the LPS group. Conversely, LFnm, LF/HF, and VLF in the LPS + VNS group decreased significantly compared with the LPS group. Hepatic NE and ACh significantly decreased within 6 h after LPS injection compared with the basal level and the control groups (P < 0.05). VNS did not significantly improve hepatic NE, but the ACh levels in the LPS + VNS group were higher than those in other groups. Sympathetic and vagal nervous systems are enhanced following endotoxemia. The overexcitation of the sympathetic system leads to sympathetic-vagal disequilibrium. The rebalance of the sympathetic and vagal system is crucial for critically ill patients.
No preview · Article · Jun 2010 · Journal of Neural Transmission
[Show abstract][Hide abstract] ABSTRACT: To determine the clinical relevance of 13 reported common single-nucleotide polymorphisms (SNPs) of cytokine genes in patients with major trauma.
Thirteen SNPs in nine key cytokine genes were selected for association study in 308 patients with major trauma on the basis of previous functional or association data. An allele-specific oligonucleotide array was developed and used to genotype 308 patients with major trauma. The clinical relevance of the 13 SNPs was assessed by observation of cytokine production and outcome of trauma patients.
Results from the allele-specific oligonucleotide array indicated that 8 [interleukin-1beta (IL-1beta)/-1470, IL-1beta/-511, IL-1beta/-31, IL-4/-589, IL-6/-572, IL-8/-251, IL-10/-819, and tumor necrosis factor alpha (TNFalpha)/-308] out of the 13 SNPs were associated with respective ex vivo cytokine production by peripheral leukocytes in response to lipopolysaccharide (LPS) stimulation at admission, or risk of development of sepsis or organ dysfunction in major trauma patients. Patients with more than four risk alleles of the eight SNPs had more than 50% sepsis morbidity and more severe organ dysfunction.
Polymorphisms of IL-1beta/-1470, IL-1beta/-511, IL-1beta/-31, IL-4/-589, IL-6/-572, IL-8/-251, IL-10/-819, and TNFalpha/-308 are susceptibility loci for the development of sepsis and organ dysfunction in major trauma patients.
No preview · Article · Mar 2010 · Intensive Care Medicine
[Show abstract][Hide abstract] ABSTRACT: Several polymorphisms in the CD14 promoter have been reported to be associated with various inflammatory diseases. However, conflicting results have been shown in association studies in different populations. This study aimed to investigate the possible functional significance of both the G-1145A and T-159C polymorphisms in the CD14 promoter and their association with organ dysfunction and sepsis in adult trauma patients.
Genetic, functional, and association studies.
National Key Laboratory of Trauma and Departments of Traumatic Surgery in two teaching hospitals.
Three hundred twenty-five healthy volunteers and 105 patients with major trauma.
Among the five single nucleotide polymorphisms identified within CD14 promoter in a Chinese Han population, two single nucleotide polymorphisms (G-1145A and T-159C) were selected according to bioinformatics analysis. Promoter activity of polymorphisms was determined using the reporter gene assay. Plasma sCD14 and tumor necrosis factor-alpha levels were measured by enzyme-linked immunosorbent assay. Both single nucleotide polymorphisms significantly reduced transcriptional activity of the promoter, and were significantly associated with a decrease of inducible sCD14 and tumor necrosis factor-alpha production in an allele-dose effect. Moreover, trauma patients carrying the -1145 A or -159 C allele appeared to have a decreased risk of multiple organ dysfunction and sepsis. In addition, both polymorphisms had a marked synergistic effect.
The CD14/-1145 and -159 polymorphisms are functional variants, which may function in a synergistic fashion, and could be used as biological risk predictors of multiorgan dysfunction and sepsis in trauma patients.
No preview · Article · Aug 2008 · Critical care medicine
[Show abstract][Hide abstract] ABSTRACT: To observe the effects of stimulation of intact vagus nerve (IVNS) on systemic inflammatory response in rats.
The model of systemic inflammatory response was reproduced by lipopolysaccharide (LPS). One hundred Sprague Dawley rats were randomly divided into A group (with intravenous injection of LPS), B group (with stimulation of efferent nerve trunk of vagus nerve after intravenous injection of lipopolysaccharide and vagotomy), C group (with stimulation of intact vagus nerve after intravenous injection of LPS), D group (with vagus nerve stimulation after injection of equivalent isotonic saline), E group (with intravenous injection of equivalent isotonic saline), with 20 rats in each group. Five rats of each group were used to determine mean aortic pressure (MAP) before injection and l0 min, 0.5 h, 1 h, 2 h, 4 h and 6 h after injection. The serum levels of tumor necrosis factor (TNF) alpha and interleukin (IL) 10 were determined with enzyme linked immunosorbent assay before injection and 2, 4 and 6 hour after injection.
The level of MAP rose in A, B, C groups at 10 min after injection, especially in A group (134.40 +/- 7.3 mm Hg, l mm Hg = 0.033 kPa, P < 0.05), but it dropped in above groups at 30 min after injection. The level of MAP in A group was obviously lower than that in B, C groups during 10 min -6 h after injection. The serum level of TNF-alpha in A group was significantly higher than that in B, C groups at 2, 4, 6 hours after injection (P < 0.05). Compared with that in C group, the serum level of IL-10 in A, B groups lowered markedly at 4, 6 hours after injection (P < 0.05).
IVNS can stabilize hemodynamics and exert have anti-inflammatory effects at early stage of systemic inflammatory response.
No preview · Article · May 2008 · Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = Chinese journal of burns
[Show abstract][Hide abstract] ABSTRACT: To investigate the functional significance of the reported single nucleotide polymorphisms (SNPs) in the promoter of the myeloid differentiation-2 (MD-2) gene.
Functional gene polymorphisms of innate immune receptors have been shown to be critical determinants of the immune inflammatory response. MD-2 is an important signaling enhancer protein in the endotoxin (LPS) receptor complex. Although a total of 156 SNPs have been identified within the whole MD-2 gene, little is known about the functional significance of these SNPs.
: The possible biosignificance of 8 reported SNPs was analyzed using on-line software tools. The selected SNPs were then genotyped using a restriction fragment length polymorphism method applied to 711 healthy Chinese volunteers. Their functional effects were assessed by the observation of transcription activity, MD-2 mRNA expression, and leukocyte response to ex vitro LPS stimulation. Moreover, the clinical relevance of these SNPs was investigated in 105 patients with major trauma.
Three SNPs (C-1625G, A-1064G, and A-475T) in the MD-2 promoter were selected based on bio-informatic analysis. Both -1625 and -1064 SNPs, rather than -475, were seen in the Chinese population, with frequencies of 19.8% (-1625G) and 34.7% (-1064G). But only the -1625 polymorphism was found to affect MD-2 promoter activity. Moreover, the expression of MD-2 mRNA and the production of TNF-alpha in whole blood leukocytes, in response to LPS stimulation, were significantly increased in subjects with the -1625 G allele. Patients who possessed the -1625 G allele were more likely to experience complications with organ dysfunction and sepsis after major trauma. All these associations were in allele-dose dependent effect.
The MD-2/-1625 polymorphism is an important functional variant.
Full-text · Article · Aug 2007 · Annals of Surgery
[Show abstract][Hide abstract] ABSTRACT: To investigate the polymorphisms of myeloid differentiation-2 (MD-2) gene promoters, and to explore whether such polymorphisms are associated with the susceptibility to multiple organ dysfunction syndrome (MODS) and sepsis in Chinese Han population.
Using polymerase chain reaction-restriction fragment length polymorphism method, the authors detected the single nucleotide polymorphisms of the promoter region of MD-2 gene at position - 1625C/G in 105 severe trauma patients (42 with sepsis). The organ function was scored.
The frequency of CC genotype in MD-2 gene promoter region at position - 1625 was 0.5 (21/42) in septic patients and 0.7 (44/63) in non-septic patients. The frequency of CG genotype was 0.38 (16/42) in septic patients and 0.27 (17/63) in non-septic patients. The frequency of GG genotype was 0.12 (5/42) in septic patients and 0.03 (2/63) in non-septic patients. The MODS scores in trauma patients carrying G allele at position - 1625 were significantly higher than those carrying C allele (P<0.001 for dominant effect, and P>0.05 for recessive effect). Moreover, trauma patients carrying G allele appeared to have higher risk of sepsis comparing to those carrying C allele (OR 0.477, 95% CI 0.266-0.855, P<0.05). Sepsis morbidity was significantly different between subjects with C and G alleles (P<0.05 for dominant effect, P>0.05 for recessive effect).
The polymorphisms of the promoter region of MD-2 gene at position - 1625 C/G is correlated with MODS and sepsis after severe trauma in Chinese Han population. The people with - 1625 G allele in the promoter region of MD-2 gene may be a risk factor of severe complications.
No preview · Article · Aug 2007 · Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae
[Show abstract][Hide abstract] ABSTRACT: Objective
To construct and identify the recombinant plasmids PPARγ-pSUPER-EGFP for RNA interference.Methods
The pSUPER-EGFP vectors were used to transcribe functional short interfering RNA (siRNA). Four pairs of 64 nt PPARγ siRNA encoding sequences were inserted into the downstream of the H1 promoter. The recombinant plasmids were confirmed by double digestion with the enzymes and sequencing. Western blotting was used to examine the silencing effect of PPARγ gene in RAW264. 7 cells. Following procedures were used to optimize the experiments: the oligonucleotides were incubated 5 min at 95 C and cooled automatically in boiled water bath to anneal, and then phosphorylated oligonucleotides, pSUPER-EGFP plasmids was digested with Bgl II and Hind II, and the product was ligated into digested pSUPER-EGFP plasmids, and transforming the ligation products followed by screening and identifying positive clones.ResultsFour kinds of positive clones producing 285 bp fragments were selected. Sequencing further proved their correctness. Four recombinant plasmids containing corresponding PPARγ gene-specific target sequences induced the silencing of its target gene more or less.Conclusion
The optimizing method in constructing these recombinant plasmids serves other plasmid-based RNA interference research. The final plasmids PPARγ-pSUPER-EGFP established the basis for research on the function of PPARγ gene.
No preview · Article · Feb 2007 · Journal of Medical Colleges of PLA