1.Proteomic analysis of the serum from patients with acute rejection after liver transplantation
Qi JIANG ; Yawei RU ; Keqiu LI ; Guang LI
Tianjin Medical Journal 2015;43(5):449-452,577
Objective To investigate the protein markers that specifically expressed in patients with acute rejection (ACR) after liver transplantation, and to explore preliminarily the mechanisms. Methods Serum samples from three patients with pathologically confirmed ACR after liver transplantation in Tianjin First Central Hospital were collected as ACR group. Three serum samples from patients with normal liver function indicators after liver transplantation were collected as No-ACR group. And six serum samples from healthy examination were mixed with equal amount as healthy control group. Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) was employed to separate, screen and identify the differentially expressed proteins between three groups. KEGG and STRING software were applied to deeply analyze the data of three groups. Results A total of 88 differentially expressed proteins were found between ACR group and healthy control group. There were 39 differentially expressed proteins between No-ACR group and healthy control group. Ten differentially expressed proteins were acquired between ACR group and No-ACR group. Comparing 88 and 10 differentially expressed proteins, 9 proteins were the same. Among 88 differentially expressed proteins, 30 of them showed a direct interaction, and can be positioned in 13 signaling pathways based on KEGG and STRING software. Fourteen (46.67%) of the 30 proteins were located in the complement and coagulation cascade pathway. Among 39 differentially expressed proteins, which were detected between No-ACR group and control group, 10 proteins showed a direct interaction including 9 proteins concentrated in the complement and coagulation cascade pathway. Conclusion By proteomic analysis, nine differentially expressed proteins are obtained, which may be regarded as the candidate bio-markers for ACR early diagnosis after liver transplantation. The complement and coagulation cascades system is significantly adjusted after liver transplantation, indicating this pathway plays an important role in the occurrence of ACR.
2.Clinical application of high resolution melting analysis in mutation detection of phenylulanine hydroxylase gene
Chao HE ; Ru LI ; Jianhui JIANG ; Can LIAO
Chinese Journal of Laboratory Medicine 2011;34(9):836-841
Objective To detect PAH gene mutations in classical PKU patients by HRM analysis. MethodsMutation scanning of PAH gene were performed in 17 classical PKU patients by HRM analysis ( LightScanner), covering the 13 exons and exon-intron boundaries. The HRM results were further confirmed by DNA sequencing, and the sensitivity and specificity of HRM method in PKU diagnosis were also evaluated. In addition, prenatal diagnosis was performed in two fetuses at risk for classical PKU. Results In the 17 patients, two mutations were identified in 16 patients, three mutations were identified in 1 patient.In this subject, a total of 22 different pathogenic mutations : 194V( c. 280A > G), IVS4nt-1 G > A( c. 442-1G > A), R158Q( c. 4736 > A), Q160X( c. 478C > T), W187X( c. 561G > A), E6nt-96A > G( c. 611A >G), G239D( c. 716G > A), R241 C( c. 721C > T), R243Q( c. 728G > A), G247R (c. 739G > C), G247V (c. 740G>T), R261X(c. 781C >T), PR261Q(c. 782G > A), H264R (c. 791A > G), F302fsX39 (c. 904delT), E305K( c. 913G > A), G312V( c. 935G > T), Y356X( c. 1068C > A ), V399V ( c. 1197A >T), R408Q(c. 1223G > A), T418P(c. 1252A > C) , A434D(c. 1301C > A), 3 silent mutations Q232Q (c. 696G > A), V245V(c. 735G > A), L385L(c. 1155C > G), and one single nucleotide polymorphism rs2280615 ( c. 402A > C) were identified, of which 194V ( c. 280A > G), Q160X ( c. 478C > T), H264R (c. 791A > G), G312V( c. 935G > T) and E305K ( c. 913G > A) were novel mutations identified in PAH gene. The prenatal diagnosis results of the two fetuses : one was diagnosed as normal, the other was diagnosed as a carrier. In this study, the sensitivity and specificity for mutation detection by HRM were 100%, and the HRM results were consistent with DNA sequencing results. Conclusions HRM analysis is a simple,accurate, rapid, high-throughput and low-cost genetic analysis approach. It could be applied to mutation scanning of classical PKU of PAH gene and rapid prenatal diagnosis in parents with known mutations.
3.Changes of Amplitude of Ion Channels Currents in Developing Hippocampal Neurons
zhen-biao, LI ; xi-ru, WU ; yu-wu, JIANG
Journal of Applied Clinical Pediatrics 2004;0(11):-
Objective To study the changes of the amplitude of ion channels currents in developing hippocampal neurons.Methods Using whole-cell patch-clamp techniques in cultured hippocampal neurons whose cultured day were 6 d and 16 d,respectively,changes of the amplitude of ion channels currents in developing hippocampal neurons were explored.Results Compared with the hippocampal neuron whose cultured day was 6 d,there were no statistical differences of the amplitude of voltage dependent sodium currents of hippocampal neuron whose cultured day was 16 d.The amplitude of voltage dependent potassium currents of hippocampal neuron whose cultured day was 16 d were significantly increased(P
4.Changes of N-Methyl-D-Aspartate-Receptor-Channels Current in Developing Hippocampal Neurons after Hypoxia and Effect of Adenosine Intervention
zhen-biao, LI ; xi-ru, WU ; yu-wu, JIANG
Journal of Applied Clinical Pediatrics 2004;0(12):-
Objective To study the changes of N-methyl-D-aspartate(NMDA)-receptor-channels current in developing hippocampal neurones during hypoxia and effect of adenosine intervention.Methods Whole-cell patch-clamp techniques cultured hippocampal neurons whose cultured day were 6 days and 16 days respectively,the amplitude of the NMDA-receptor-channels currents of hippocampal neuron were determined.And the effect of hypoxia on the NMDA-receptor-channels current,and adenosine regulatory mechanisms in cultured hippocampal neurons were explored.Results During hypoxia,compared with control group,the amplitude of the NMDA-receptorchannels currents of hippocampal neuron whose cultured day was 6 days were significantly increased(P
5.Asymmetric Morita-Baylis-Hillman Reaction of Conjugated Nitroalkene with Activated Aldehyde Catalyzed by Cinchona Alkaloids
Lingyu SUN ; Da LI ; Zhongjing HE ; Wei YAO ; Ru JIANG
China Pharmacist 2017;20(2):216-220
Objective:To develop a catalytic system for the asymmetric Morita-Baylis-Hillman ( MBH) reaction of conjugated ni-troalkene with activated aldehyde, and screen out the chiral catalysts with high activity and enantioselectivity. Methods: Totally 21 chiral organocatalysts were applied in the asymmetric MBH reaction ofβ-nitrostyrene with ethyl glyoxylate, and the ee value was deter-mined by chiral HPLC. The effects of temperature, solvent and substrate ratio on the catalytic reaction were investigated. Results: In the presence of cinchona alkaloid catalyst (DHQ)2AQN, β-nitrostyrene reacted with ethyl glyoxylate in toluene at 0℃ affording the MBH adduct in 60% yield with good enantioselectivity (up to 56.9% ee). Conclusion: The bis-cinchona alkaloids with aromatic bridging group are the efficient catalysts for the asymmetric MBH reaction ofβ-nitrostyrene with ethyl glyoxylate, and moderate isolated yield and enantioselectivity are obtained.
6.Study of the Growth Rate and Comparative Analysis of Esterase Isoenzyme from Two Isolates of Polyporus umbellatus
Guang-Bo XU ; Yan-Ru LI ; Tai-Yuan LI ; Yun-Jiang LIANG ; Wei-Jie FU ;
Microbiology 1992;0(03):-
Comparative analysis of characteristic of species and esterase-isoenzyme of isolates of Polyporus umbel-latus from different regions were processed. The results indicated that isolates of Jizhaoling ( Z) and Zhushiling (ZJ) have significant differences in characteristic, and enzymatic band types of the two species also have significant differences. The homology at genetics between the two isolates is 0% , and consanguinity between the two i-solates is the farthest.
7.Gene detection of vancomycin-resistant Enterococcus isolated from a teaching hospital in Shanghai
Ru LI ; Xiaogang XU ; Min LI ; Xiaofei JIANG ; Quhao WEI ; Yuan Lü
Chinese Journal of Laboratory Medicine 2010;33(5):430-434
Objective To investigate the prevalence of VRE in Huashan hospital of Shanghai from 2007 to 2009, and to examine the molecular characteristics of the VRE isolates.Methods A total of 890 non-repetive clinical isolates of Enterococcus were screened by the agar screening method ( ADSP method).Broth dilution susceptibility test was performed to determine the antimicrobial susceptibility of Enterococcus isolates to vancomycin and teicoplanin.The resistant genes and virulent genes of VRE isolates were investigated by PCR and sequencing methods.VRE isolates were classified by MLST and six isolates of VRE from 2007 to 2008 were analyzed by PFGE.Results Thirteen VRE isolates were identified by ADSP method and broth dilution susceptibility test. Six of them were resistant to vancomycin but sensitive to teicoplanin ( vancomycin MICs were from 64 to 256 μg/ml).The sequencing data of PCR products indicated these isolates might harbor a potential novel vancomycin resistant gene, which was different from the one reported in previous studies. The rest 7 isolates harbored vanA gene. The MICs of these isolates to vancomycin and teicoplanin were 32 - 64 μg/ml and 16 - 32 μg/ml, respectively.MLST results revealed 4 STs were identified in 13 VRE isolates.Eleven isolates belonged to clonal complexes(CC) 17.The positive rates of esp gene and hyl gene were 69.2% and 30.8%, respectively.Conclusions This study suggests that the most common VRE clone in Huashan Hospital was CC17.A potentially novel vancomycin resistance gene was identified, and further work needs to be done to investgate the function and the location of this novel gene.
8.Botulinum toxin type A and ethyl alcohol for treating lower extremity spasticity after stroke
Jiang LI ; Ruyi LI ; Chenhan WANG ; Ru ZHANG ; Yongxiang ZHANG ; Sishan GAO ; Qiang WANG
Chinese Journal of Physical Medicine and Rehabilitation 2016;38(7):504-508
Objective To compare the effectiveness of botulinum toxin type A ( BTXA) and ethyl alcohol ( EA) in treating lower extremity spasticity after stroke. Methods This was a randomized, case-control study. A to-tal of 92 eligible stroke survivors completed the study. They were randomly divided into a BTXA group of 48 and an EA group of 44 according to a random number table. The gastrocnemius, soleus and posterior tibial muscles of the af-fected limb were chosen as injection sites. The BTXA group was injected with 50 to 200 IU of BTXA ( at 50 U/ml) at one to four sites in each muscle, with a total injection dose of less than 600 U. The EA group was injected with less than 10 ml of 50% EA (0.1 to 0.5 ml at each site). Before and 2, 4 and 12 weeks after the injection, both groups were evaluated using the modified Ashworth scale (MAS), a 3 m timed up and go test (TUG), a timed 10 meter walk ( 10m-WT) and each was asked to assess their pain level using a visual analogue scale ( VAS) . Any adverse re-actions were also observed. Results Two weeks after the injection, the average MAS score of both groups had im-proved significantly compared to that before the injection. The average improvement in the BTXA group was signifi-cantly less than in the EA group. No significant differences were found in other measurements. After four weeks the average MAS score of the BTXA group was still significantly different from that before injection or from 2 weeks previ-ously, but the EA group now showed no significant difference from before the injection. The average TUG, 10m-WT and VAS scores of both groups had improved significantly compared to those of the earlier time points. Twelve weeks after the injection, the average MAS, TUG, 10m-WT and VAS scores of the BTXA were still significantly improved compared to before the injection, but in the EA group only the average score VAS reading was significantly improved. There were then significant differences between the two groups in all of the measurements. Conclusions Both BTXA and EA can relieve muscle spasticity. Both take effect within 2 weeks, but the former has fewer side effects than the latter and a longer duration of therapeutic effect.
9.Indomethacin for prevention of post-ERCP pancreatitis and hyperamylasemia
Tao LIN ; Yi ZHOU ; Jun WANG ; Ru YAO ; Li JIANG ; Rutang FANG ; Suli WANG ; Yan XIE
Chinese Journal of Digestive Endoscopy 2012;29(4):185-187
Objective To investigate the preventive effect of Indomethacin for post-ERCP pancreatitis and hyperamylasemia.Methods A total of 600 patients,who were undergoing ERCP,were randomly divided into 3 groups to receive anal Indomethacin (n=200),intravenous octreotide (n=200) or no special medication (n=200) before ERCP.The level of serum amylase before and 24h after ERCP were measured,and the rate of acute pancreatitis and hyperamylasemia after ERCP were assessed.Results Serum amylase levels before ERCP of all groups were normal.The mean serum amylase level of Indomethacin group (101.3±77.7 U/L) after ERCP was significantly lower than those of octreotide group ( 176.6±138.3 U/L,P =0.040 ]and control group (227.2±264.9 U/L,P=0.048),while there was no difference between octreotide group and control group ( P>0.05 ).The incidence of post-ERCP pancreatitis in Indomethacin group (2.5%) was significantly lower than that of control group (9.5%,P=0.003),while there was no difference between octreotide group (4.5%) and control group ( P=0.05 ).The incidence of hyperamylasemia after ERCP in Indomethacin group (5.5%) was significantly lower than that of control group ( 13.5%,P=0.006 ),while there was no difference between octreotide group (10.0%) and control group ( P>0.05 ).Conctusion Anal administration of Indomethacin before ERCP can effectively reduce the incidence of acute pancreatitis and hyperamylasemia after ERCP.
10.VOCC and BKCa mRNA expression in kidney tissues of IgA nephropathy patients
ming, LI ; geng-ru, JIANG ; min-yi, CHEN ; xiu-ying, LIU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(10):-
0.05). ConclusionThe expression of VOCC mRNA and BKCa mRNA in kidney tissues of IgA nephropathy patients are abnormal.There is positive correlation between the abnormal expression of VOCC mRNA and BKCa mRNA and total glomerular pathological lesions integrals.The expression of VOCC mRNA and BKCa mRNA in kidney tissues of IgA nephropathy may serve as the indictor for the disease progression.