1.The relevance among serum CRP level,CRP gene C+1444T polymorphism and the risk with acute myocardial infarction *
Haitao YANG ; Guoping HE ; Chuanping QI ; Lianhong XU ; Yichao QIAN
Chongqing Medicine 2013;(27):3222-3224
Objective To investigate the relationship between serum C-reactive protein(CRP) level ,CRP gene C+1444T poly-morphism and the risk with Acute myocardial infarction (AMI) in Chinese Han population in Sunan region .Methods The CRP gene C+1444T polymorphism was genotyped by Polymerase reaction restriction-fragment length polymorphism (PCR-RFLP) and the serum CRP level was measured by enzyme linked immunosorbent assay (ELISA) between 227 patients with AMI(AMI group) and 161 control subjects .Results No differences were found in genotype distribution between AMI group and controls (CC 82 .38% ,CT 17 .62% ,TT 0% vs 86 .96% ,13 .04% ,0% )(P>0 .05) .The serum CRP level in AMI group was significantly higher than controls(P< 0 .01) .There was no differences in the serum level between any genotypes of the CRP gene C + 1444T (P<0 .05) .Conclusion The CRP gene C+1444T polymorphism is not associate with increased risk of AMI ,and it have no effect with the serum level in Chinese Han population in Sunan region .
2.Protective effect of aqueous extract from dioscorea nipponica Makino against carbon tetrachloride-induced acute liver injury in mice via TLR4/MyD88 signal pathway
Liping LIU ; Lianhong YIN ; Xufeng TAO ; Lina XU
International Journal of Traditional Chinese Medicine 2017;39(9):815-819
Objective To investigate the effect of the aqueous extracts from Dioscorea nipponica Makino (AEDN) against the carbon tetrachloride (CCl4)-induced mice acute liver injury by regulating TLR4/MyD88 signal pathway.Methods 60 mice were randomly divided into control group, model group, low, medium and high dose AEDN groups according to radom number table with 10 mice in each group. Mice in low, medium and high dose AEDN groups were adiminstrated with 50, 100 and 200 mg/kg AEDN, in control and model groups were adiminstrated with solvent once a day for 7 consecutive days. Two hours after the last administration, mice were intraperitoneal injected with with 0.3% CCl4 olive oil solution to induce acute liver injury model, except for the mice in control group. Twenty-four hours after injection, the expressions of TLR4, MyD88 and NF-κB in liver tissue were evaluated by Western blot, mRNA levels were evaluated by PCR, and the AST and ALT levels in serum were also detected.Results Compared with model group, the serum AST (98.00 ± 17.75 U/L, 57.49 ± 9.66 U/L, 39.60 ± 9.49 U/Lvs. 113.40 ± 9.71 U/L) and ALT levels (76.00 ± 14.73 U/L, 50.70 ± 9.35 U/L, 35.25 ± 9.93 U/Lvs. 95.42 ± 11.64 U/L) were significantly decreased in low, medium and high dose AEDN groups (P<0.01); MyD88 (0.67 ± 0.21vs. 1.74 ± 0.42), NF-κB p65 (0.51 ± 0.09vs. 1.76 ± 0.31) and TLR4 (0.97 ± 0.25vs. 2.99 ± 0.72) levels were down-regulated in high dose AEDN group (P<0.01); the mRNA levels of IL-6 (2.22 ± 0.25, 1.76 ± 0.31vs. 5.20 ± 0.60), IL-1β (1.96 ± 0.35, 1.47 ± 0.23vs. 7.37 ± 0.99)、TNF-α (2.06 ± 0.25, 1.34 ± 0.33vs. 2.98 ± 0.50) in medium and high dose AEDN groups significantly decresed (P<0.01).Conclusions The AEDN has protective effect against CCl4-induced acute liver injury in mice via adjusting TLR4/MyD88 signal pathway.
3.Establishment and application of dual real-time fluorescent RT-PCR method for detection of Enterovirus
Lianhong XU ; Yulin YUE ; Yongfang WANG ; Ying CHU ; Lixin JIANG
Chongqing Medicine 2016;45(33):4688-4690,4741
Objective To develop a dual real-time fluorescent RT-PCR method for rapid detection of enterovirus(EV)and en terovirus type 71(EV71).Methods Specific primers and probes were designed and the dual real-time fluorescent RT-PCR reaction system was established.The quantitative standard curve was drawn;its sensitivity and precision were evaluated.Feces and throat swab specimens of 109 clinical patients with hand foot and mouth disease were collected and tested by using this method.Then the obtained results were compared with those detected by commercial EV71 PCR kit.Results The relative coefficient(2)of EV and EV71 standard curve established by the dual real-time fluorescent RT-PCR method were both 0.998.Its sensitivity reached 0.5 TCID50/mL for detecting EV and 0.05 TCID50/mL for detecting EV71.The within-run precision for detecting EV and EV71 was <3% and total precision≤4%.The results showed good specificity for the detection of enterovirus and non-enterovirus.In 109 detected clinical samples,84 cases of EV positive samples were detected,in which 56 cases were EV71 positive with the total positive rate of 51.4 %,which was consistent with the result of simple fluorescent RT-PCR commercialization kit(P=1.000).Conclusion The established dual real-time fluorescent RT-PCR method has high sensitivity and good stability,which has an important significance for early high throughput rapid diagnosis of hand foot and mouth disease.
4.Relationship of the phosphodiesterase 4D gene rs153031 polymorphism and the susceptibility of unstable angina pectoris
Hao XU ; Guoping HE ; Chuanping QI ; Lianhong XU ; Haitao YANG ; Yichao QIAN
Chongqing Medicine 2014;(5):551-552,555
Objective To investigate the association between the phosphodiesterase 4D(PDE4D) gene rs153031 polymorphism and the susceptibility of unstable angina pectoris(UAP) in Chinese Han population of Changwu region .Methods The PDE4D gene rs153031 polymorphism was genotyped by Taqman probe in 172 UAP patients(UAP group) ,as well as in gender-and-age-matched 220 subjects without coronary heart disease(CHD)(control group) .Results In this crowd ,there was PDE4D gene rs153031 poly-morphism in patients with UAP and in subjects without CHD .Compared with control group ,frequencies of GG ,GA ,AA genotypes and G allele of rs153031 in UAP group showed no statistical differences (P> 0 .05) .Conclusion In Chinese Han population of Changwu region ,PDE4D gene rs153031 polymorphism shows no association with the susceptibility of UAP .
5.Study of Plasma amino acid related metabolites of septic rats using gas chromatography-mass spectrometry
Jin SU ; Yimin ZHU ; Yu JIANG ; Lianhong ZOU ; Xiaoliang LIU ; Yixiao XU
Chinese Critical Care Medicine 2017;29(4):332-336
Objective To reveal the pathophysiological changes of sepsis, the plasma metabonomics of septic rats was determined and differential metabolites were determined by gas chromatography-mass spectrometry (GC-MS). Methods Male Spraguee-Dawley (SD) rats about 8 weeks were randomly divided into sham group (n = 18) and sepsis group (n = 24). Cecal ligation and puncture (CLP) was used to build sepsis model, while cecum was kept intact only in the sham group. 6 hours after the operation, rats were anesthetized, and blood was harvested through heart thoracotomy. Then the plasma metabonomics was detected by GC-MS and metabolic profile analysis was performed to find the relative differential metabolites.Results Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) showed that the metabolic profiling of the sepsis group was significantly different from the sham group. 259 kinds of metabolites were got by GC-MS, and 69 kinds of differential metabolites were found between sham group and sepsis group, in which 23 differential metabolites were related to amino acid metabolism. Compared with sham group, the contents of putrescine, N-glutamic acid, hydroxynorvaline, 3-cyanuric acid, D-alanyl-D-alanine and urea in the plasma of septic rats increased significantly, which ratios of sepsis/sham group were 10.876, 6.394, 2.800, 2.226, 1.323, 1.203, respectively (allP < 0.05). On the other hand, the contents of oxygen generation of proline, citrulline, glutamine, su-beta-hydroxy aspartic acid, citric acid, N-methyl-DL-alanine, serine, lysine, threonine, N-formyl-L-methionine, methionine, alanine, nicotinuric acid, N-methyl-L-glutamic acid, trans-4-hydroxy-L-proline, proline, L-glutamic acid in the plasma of septic rats decreased significantly, which ratios of sepsis/sham group were 0.858, 0.853, 0.834, 0.816, 0.816, 0.814, 0.813, 0.801, 0.793, 0.792, 0.774, 0.766, 0.748, 0.727, 0.716, 0.674, 0.603, respectively (allP < 0.05).Conclusions Through the GC-MS analysis of plasma metabonomics of septic rats, we found abnormal energy metabolism changes. The content of amino acid in plasma might be a method to evaluate the energy metabolism status of sepsis.
6.Analysis of screening and therapeutic effect of congenital hypothyroidism in Zhongshan
Lianhong HUANG ; Yumei SHANGGUAN ; Simao FU ; Yuling LIU ; Junbin OU ; Kang XU ; Cuimei ZHANG
Journal of Clinical Pediatrics 2014;(9):876-880
Objectives To summarize screening and therapeutic effects of congenital hypothyroidism (CH) in Zhongshan. Methods The thyroid stimulating hormone (TSH) concentration in dried heel blood samples on iflter paper was detected using time-resolved fluorescence immunoassay. The cases of positive screening tests were called back for further examination of venous blood TSH concentration using chemiluminescence method. Fifty-four children with permanent CH treated routinely for 2 years (CH group) and 120 age-gender matched health children (control group) were recruited. The physical development (height, body weight) was monitored. The neurodevelopment and temperament type were tested using Pediatric Nneuropsychological Development Assessment and Children's Temperament Scale respectively at 6 and 24 months after birth. Results Two hundred eight-five thousand two hundred forty-two neonates were screened. One hundred and forty cases were confirmed and the incidence rate was 1/2037. There was no statistical difference in length-for-age z score (LAZ) and weight-for-age z score (WAZ) between CH and control group (P>0.05). The neurodevelopment in CH group was normal, but gross motor development was worse than that in control group (P<0.05). The temperament type and distribution had statistical difference between CH and control group (P<0.05). The percentage of the dififcult type and central dififcult type was increased in CH group as compared to control group, especially in the activity, adaptability, reaction intensity and perseverance (P<0.05). Conclusions The physical and neurodevelopment are nearly normal in patients with CH after early supplementation, but the psychological behavior problems need to be focused on in the process of intervention.
7.Efficacy of pregabalin in patients with painful radiation-induced brachial plexus injury
Jianhua YANG ; Lianhong YANG ; Rong WU ; Yongteng XU ; Jinping CHENG ; Yamei TANG
Chinese Journal of Radiological Medicine and Protection 2015;35(12):925-928
Objective To evaluate the efficacy of pregabalin on painful radiation-induced brachial plexus injury.Methods A total of forty-five patients with painful radiation-induced brachial plexus injury were assigned to treatment group with pregabalin 150-600 mg/d combined with conventional therapy, and to control group with conventional treatment for 12 weeks.The visual analogue scale (VAS) and neuropathic pain symptom inventory (NPSI) were evaluated.The response rate (proportion of subjects with at least a 30% to 50% reduction in visual analogue scale) was evaluated as the primary endpoint.Results The response rate of the pregabalin group(36.4%) was significantly higher than that of the control group (9.1%) (x2 =4.98, P < 0.05).Although hospital anxiety and depression level, clinical global impression of change, and clinical global impression of change (CGIC) did not differ significantly between pregabalin group and control group (P > 0.05), improvements in neuropathic pain symptom inventory (NPSI), patient global impression of change (PGIC) and the sleep interference score(t =-6.62,-5.65,-6.16, P < 0.05)suggested some utility in the management of radiation-induced brachial plexus.Conclusions Pregabalin treatment combined with conventional treatment exerts efficacy on patients with painful radiation-induced brachial plexus and patients are well tolerated.
8.Influence of simulated microgravity on rat islets morphology in culture
Chun SONG ; Xiuqing DUAN ; Yeqing SUN ; Xi LI ; Liou HAN ; Ping XU ; Chunfang SONG ; Lianhong JIN
Chinese Journal of General Surgery 2001;0(07):-
Objective To observe the influence of simulated microgravity on rat islet. Methods Isolated islet were assigned to flask-culture or bioreactor-culture. Gross structure and ultrastructure of islet were observed by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Results Islets ultrastructure on 7th day in bioreactor closely resembled fresh islets,with well-formed secretory granules and abundant mitochondria. SEM showed under microgravity islets communicating each other with cavity-like areas. Conclusions The ultrastructure of islets cultured under microgravity closely resembled fresh islets.
9.Transgenic mouse fetus generated from embryonic stem cells by tetraploid embryo complementation
Yanning XU ; Na GUAN ; Zhiyan SHAN ; Jingling SHEN ; Lianhong JIN ; Lei LEI
Acta Anatomica Sinica 2009;40(6):943-947
Objective To use tetraploid embryo complementation combined with gene transfer to produce genetically modified embryonic stem cells (EsCs) clones. Methods In this study, EGFP was introduced into ESCs by electroporation, and transfected positive cells were selected by G418 resistance. The tetraploid embryos were obtained from diploid blastomere electrofusion which preformed at 2-cell stage. Afterwards, 19-21 EGFP-ESCs were inserted into each tetraploid blastocyst cavity by piezo drilled microinjection,then the injected blastocysts were transferred into the uterus of pseudo-pregnancy at 2.5-day or the oviduct of 0.5-day female mice. Results The transfected ESCs maintained normal karyotype even after long-term passage (2n=40). The rate of fusion was 95.07%, and the developmental rate of tetraploid blastocyst was 95%.Totally 410 injected blastocysts were obtained. Unfortunately, we have not got any vital offsprings, except 151 implantation sites (pseudo-pregnancy 2.5 days:29.41%;the oviduct of half one day:64.37%). Furthermore, scattered EGFP expressions in transgenic fetus were observed under invert fluorescent microscope. Conclusion The transfected ESCs were observed in transgenic fetus, and the implantation rate in oviduct was higher than that in uterine.
10.Correlation between polymorphism of monocyte chemoatt-ractant protein-1 gene A-2518G single nucleotide and acute coronary syndrome
Ganwei SHI ; Guoping HE ; Gaojun CAI ; Chuanping QI ; Lei GAO ; Meng QI ; Dandan SHEN ; Zhihong QIAN ; Lianhong XU
Chinese Journal of Emergency Medicine 2012;21(7):687-693
Objective To investigate the possible correlation between the monocyte chemoattractant protein-1 ( MCP-1 ) gene A-2518G single nucleotide polymorphism (SNPs) in the promoter region and acute coronary syndrome (ACS) in Chinese Han ethnic population of Sunan region,Methods This study was conducted with a case-control design in 484 ACS patients including 290 acute myocardial infarction (AMI)patients and 194 patients with unstable angina pectoris (UAP) and 346 control subjects ruled out coronary disease by coronary angiography (control group),including 166 patients with coronary atherosclerosis and 180 subjects without coronary stenosis.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for the detection of the A-2518G polymorphism in MCP-1 gene,and then thefrequency of genetype was statistically analyzed.Results There were AA,AG and GG genotypes of MCP-1 gene A-2518G polymorphism in the ACS group and control group.The two groups could be considered as a genetic equilibrium representative by Hardy-Weinberg equilibrium ( P > 0.05 ).Compared with the control group,the frequencies of AA genotype ( 15.32% vs.16.12% ),AG genotype (53.47% vs.51.86% ),GG genotype (31.21% vs.32.02% ) and G allele genotype (57.95% vs.57.95% ) in ACS group were not significantly different ( P was 0.083,0.673,0.821 and 1.00,respectively).Multivariate logistic regression analysis indicated that there was no significant correlation between MCP-1 gene A-2518G polymorphism and ACS regardless of differences in gender,age,smoking,diabetes,TG and LDL-C ( P >0.05 ).There was no significant difference in gender and age of ACS onset between two groups ( P > 0.05).There were no significant differences in the frequencies of AA,AG and GG genotypes and G allele genotype among AMI group,UAP group and normal coronary group ( P > 0.05).Conclusions The data shows that MCP-1 gene A-2518G polymorphism is not associated with the risk of ACS in the Chinese Han ethnic population living in Sunan region.