1.Effect of homocysteine on the intestinal permeability by regulating MEK-ERK-MLCK signal transduction in experimental colitis rats
Shaozhen DING ; Hao DING ; Qiao MEI ; Xiaochang LIU ; Jing HU ; Yongmei HU ; Jianming XU
Chinese Pharmacological Bulletin 2016;32(4):498-502
Aim To investigate whether Hcy influenced the intestinal mucosal permeability by regulating MEK-ERK-MLCK pathway. Methods SD rats were divided into 4 groups:normal group, normal+Hcy group, TN-BS/ethanol group, TNBS/ethanol+Hcy group. Experi-mental colitis model with hyperhomocystinemia was es-tablished in rats with intracolonic administration of TN-BS and subcutaneous injection of Hcy. The colonic mucosal tissue was collected for histopathological exam-ination and activity of myeloperoxidase ( MPO ) . The protein expression of MLCK, p-MLCK, MEK, ERK and p-ERK in intestinal mucosal tissues was examined by Western blot method. The mRNA expression of ML-CK was examined by RT-qPCR method. Result Com-pared with the normal group and TNBS group, the DAI and HI scores and the MPO activity were increased in TNBS/ethanol+Hcy group ( P <0. 01 ) . Western blot and RT-qPCR showed that expression of MLCK, p-ML-CK, MEK, ERK and p-ERK increased in small intes-tine in TNBS/ethanol+Hcy group. Conclusion Hcy can increase intestinal permeability in TNBS-induced colitis rats by regulating the expression of MEK-ERK-MLCK signal pathway.
2.Adjuvant effect of CTA1-DD/IgG on immune responses to mode antigen ovalbu-min
Wenjiong XU ; Yan WANG ; Xuan WANG ; Mengkai QIAO ; Min DONG ; Min HE ; Limin SHI ; Xiaoxiao DONG ; Shifeng YUN ; Jie DING
Chinese Journal of Immunology 2014;(10):1349-1352
Objective:To determine the immunological effects of a new type adjuvant CTA 1-DD/IgG.Methods:Intranasal im-munization of BALB/c mice with OVA in combination with adjuvant CTA 1-DD or CTA1-DD/IgG.Blood samples were subjected to OVA-specific IgG Abs detection and IgG subclass profiles assessment using ELISA.Splenic plasma cells secreting anti-OVA Abs were detected by ELISPOT.Results:CTA1-DD/IgG showed stronger activity in inducing OVA-specific IgG Abs and splenic Abs-secreting cells compared with CTA1-DD.The enhancement of IgG1,IgG2a and IgG2b Abs were observed,and the ratio IgG2a/IgG1 were >1 in both groups.Conclusion: CTA1-DD/IgG can exert enhanced adjuvant effects compared with CTA 1-DD.Mixed IgG subclasses are induced,indicating a more balanced Th1/Th2 response.
3.Quantitative identification of TCM Q-markers based on analytical hierarchy and the entropy weight comprehensive method by taking Shaoyao Gancao decoction as an example
Li-mei FENG ; Yan-yan CHEN ; Shi-jun YUE ; Ding-qiao XU ; Rui-jia FU ; Jie YANG ; Yu-ping TANG
Acta Pharmaceutica Sinica 2021;56(1):296-305
The quality markers (Q-markers) of traditional Chinese medicine (TCM) have become a topic of interest in TCM research in recent years. Nonetheless, there is still no consensus on how to scientifically characterize TCM Q-markers. Our study establishes an identification method for TCM Q-markers based on the analytical hierarchy process (AHP) and the entropy weight comprehensive method. By constructing an evaluation system encompassing the target layer, the factor layer and the control layer, AHP can be used to analyze the weight of three core TCM quality attributes, including effectiveness, testability and specificity. Following that, the entropy weight method is employed to analyze the specific indicators for each attribute based on the literature and experimental data. Finally, the comprehensive weight of each index is obtained by combining the two weights, and the comprehensive weight and the specific value of each component is multiplied and summed to obtain the integrated score ranking, and thereby identify the TCM Q-markers. Taking Shaoyao Gancao decoction as an example, the analysis revealed that the top 8 components are as follows: paeoniflorin > quercetin > albiflorin > glycyrrhizic acid > naringenin > liquiritin > oxypaeoniflorin > benzoylpaeoniflorin, and can be identified as Q-markers of Shaoyao Gancao decoction. This study not only provides support for the establishment of quality standards and process quality control of TCM formulae, but also provides innovative ideas and methods for quantitative evaluation and accurate identification of TCM Q-markers.
4.Expression of a novel human retroviral NP9 gene and potential roles of its protein in systemic lupus erythematosus patients.
Yue-zhong REN ; Qiao-ding DAI ; Rong-zhen XU
Chinese Journal of Medical Genetics 2005;22(3):248-250
OBJECTIVETo investigate the expression of a novel retroviral (NP9) gene transcripts and the possible role of its protein in systemic lupus erythematosus (SLE) patients.
METHODSThe retroviral NP9 gene in SLE patients was isolated and cloned using RT-PCR and TA cloning techniques, and it was analyzed by sequencing. The expression of the NP9 genes in 40 patients with SLE and 48 normal controls using RT-PCR was detected. NCBI BLAST and DNASIS 3.1 software were used to analyze the features of protein of NP9 gene.
RESULTSThe positive ratio (77.5%) of the mRNA expression of the retroviral NP9 gene in SLE patients is significantly higher than that (8.3%) in normal subjects (P<0.01). The recombinant NP9 protein comprises 74 AA with pI 9.59. Amino acid sequence analysis indicates that the retroviral NP9 protein shares higher homologies with several human proteins with important biological functions.
CONCLUSIONSLE patients possess specific novel retroviral NP9 transcripts. The expression of the retroviral NP9 gene may involve in the genesis or development of SLE.
Amino Acid Sequence ; Computational Biology ; Endogenous Retroviruses ; genetics ; metabolism ; Humans ; Lupus Erythematosus, Systemic ; genetics ; physiopathology ; virology ; Molecular Sequence Data ; Retroviridae Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction
5. The function of NLRP1 in noninfectious pulmonary injury following allogeneic hematopoietic stem cell transplantation
Mingfeng LI ; Wen LI ; Lan DING ; Yulu WU ; Lu LIU ; Wen JU ; Jianlin QIAO ; Kailin XU ; Lingyu ZENG
Chinese Journal of Hematology 2017;38(7):607-611
Objective:
To explore the function of NLRP1 in noninfectious pulmonary injury (nonIPI) after allogeneic stem cell transplantation (allo-HSCT) .
Methods:
In this study, we established the model of allo-HSCT with C57BL/6 and NLRP-/- mouse as recipients. Chimera rate was measured by flow cytometry. The HE staining was used to observe the pathology changes in the lungs. NLRP1 and relevant inflammatory proteins were measured by Western Blot.
Results:
On the day 14 after allo-HSCT, the chimera rate was more than 96%, HSCs of donors had been successfully transplanted into recipients. HE staining showed that nonIPI occurred after allo-HSCT. The degrees of injuries reached the peak on day 21. In addition, the expressions of MPO, NLRP1, p20, Mature-IL-1β and Mature-IL-18 had same tends with the degrees of nonIPI. When we knocked out NLRP1 gene of recipients, the degrees of nonIPI reduced and the expressions of MPO, p20, Mature-IL-1β and Mature-IL-18 were less than in non-knockout group.
Conclusion
allo-HSCT could cause nonIPI and high expressions of MPO, p20, IL-1β, IL-18, NLRP1. Knocking out NLRP1 gene could alleviate the degrees of nonIPI and reduce the expressions of relevant inflammatory proteins, indicating that NLRP1 might be one of factors contributed to nonIPI after allo-HSCT.
6.Molecular cloning of a novel retroviral NP9 gene of human endogenous retrovirus and its viral protein expression.
Yue-zhong REN ; Qiao-ding DAI ; Rong-zhen XU
Journal of Zhejiang University. Medical sciences 2005;34(4):361-371
OBJECTIVETo investigate the relationship between retroviruses and autoimmune diseases, to clone the novel retroviral NP9 gene from human endogenous retrovirus (HERV), and to construct its expression vector.
METHODSThe viral NP9 gene was amplified and cloned by RT-PCR and T-A clone techniques, and its sequence was determined with Perkin-Elmer 377 DNA Sequencer. The amplified viral NP9 gene was subcloned into the prokaryotic express vector pQE30. The recombinant plasmids were identified by restriction endonuclease digestion and sequencing. The recombinant pQE30-NP9 protein was expressed in M15 host cells under the IPTG induction and showed with SDS-PAGE,and the corresponding NP9 viral protein was identified with Western blot analysis.
RESULTA specific band of 250 bp was amplified using RT-PCR from total RNA of peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE) and confirmed as the NP9 gene via T-A clone and DNA sequencing analyses. SDS-PAGE profile showed a clear protein band with a relative molecular weight 9 kD in the IPTG-induced samples, which was confirmed as viral NP9 protein by Western blot analysis.
CONCLUSIONThe NP9 gene has been successfully isolated and cloned from PBMCs of SLE patients and the corresponding NP9 viral protein expressed in prokaryotic expression vector.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Gene Products, env ; biosynthesis ; genetics ; Genetic Vectors ; Humans ; Lupus Erythematosus, Systemic ; genetics ; virology ; Molecular Sequence Data ; Retroviridae ; genetics ; metabolism ; Retroviridae Proteins ; biosynthesis ; genetics
7.Case-control study on needle-knife to cut off the medial branch of the lumbar posterior ramus under C-arm guiding for the treatment of low back pain caused by lumbar facet osteoarthritis.
Di LU ; Wei-xing XU ; Wei-Guo DING ; Qiao-Feng GUO ; Gou-ping MA ; Wei-min ZHU
China Journal of Orthopaedics and Traumatology 2013;26(3):214-217
OBJECTIVETo study the clinical efficacy of needle-knife to cut off the medial branch of the lumbar posterior ramus under C-arm guiding to treat low back pain caused by lumbar facet osteoarthritis.
METHODSFrom July 2009 to June 2011, 60 patients with low back pain caused by lumbar facet osteoarthritis were reviewed,including 34 males and 26 females, ranging in age from 39 to 73 years old,averaged 61.9 years old; the duration of the disease ranged from 6 to 120 months, with a mean of 18.9 months. All the patients were divided into two groups, 30 patients (18 males and 12 females, ranging in age from 39 to 71 years old, needle-knife group) were treated with needle-knife to cut off medial branch of the lumbar posterior ramus under C -arm guiding and the other 30 patients(16 males and 14 females, ranging in age from 41 to 73 years old, hormone injection group) were treated with hormone injection in lumbar facet joint under C-arm guiding. The preoperative JOA scores and the scores at the 1st, 12th and 26th weeks after treatment were analyzed.
RESULTSBefore treatment,the JOA scores between the two groups had no significant difference (P= 0.479); after 1 week of treatment, the JOA scores between the two groups had significant difference (P= 0.040), the improvement rate of hormone injection group was superior than that of the needle-knife group,which were (58.73+/-18.20)% in needle-knife group and (71.10+/-22.19)% in hormone injection group; after 12 weeks of treatment, the JOA scores between the two groups had no significant difference(P=0.569), and the improvement rate between the two groups had no significant difference,which were (50.09+/-19.33)% in the needle-knife group and (48.70+/-18.36)%) in the hormone injection group; after 26 weeks of treatment,the JOA scores between the two groups had significant difference (P=0.000), the improvement rate of hormone injection group was superior than that of the needle-knife group,which were (48.56+/-28.24)% in needle-knife group and (15.62+/-11.23 )% in hormone injection group.
CONCLUSIONUsing needle-knife to cut off the medial branch of the lumbar posterior ramus could get longer efficacy than hormone injection in the treatment of lumbar facet osteoarthritis.
Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Low Back Pain ; etiology ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Osteoarthritis, Spine ; complications ; surgery ; Spinal Nerves ; surgery
8.Phylogenetic Analyses and Characterization of the VP7 Genes of Serotype G9 Rotaviruses Circulating in Nanjing (China), 2012-2013.
Xuan WANG ; Limin SHI ; Mengkai QIAO ; Yan WANG ; Min HE ; Wei YONG ; Xuefei DU ; Baofu GUO ; Guoxiang XIE ; Ziqian XU ; Jie DING
Chinese Journal of Virology 2015;31(4):425-432
Group-A rotaviruses are recognized as the most common cause of acute diarrhea. Phylogenetic analyses of the VP7 genes of rotaviruses circulating in Nanjing (China) could aid in the development of rotavirus vaccines. A total of 908 stool specimens were collected from patients suffering from acute diarrhea in Nanjing between October 2012 and December 2013, and were tested further for rotaviruses. Fifty rotavirus isolates selected randomly were typed by reverse transcription-polymerase chain reaction using serotype-specific primers for G genotyping. VP7 genes of 19 G9 strains were sequenced for further genetic characterization. Among the 908 stool specimens examined during the surveillance period, 103 (11.34%) were rotavirus-positive. G9 was the most predominant genotype (78.0%), followed by G2, G1 and G3. Sequence and phylogenetic analyses of the VP7 genes of serotype G9 rotaviruses revealed these strains to comprise two lineages (G9-VI, G9-III) and to be dominated by the G9-VI lineage (which belonged to a unique subcluster of Japanese and Chinese G9 strains). Amino-acid sequences of the four antigenic regions (A, B, C or F) were variant among a portion of strains, which may have contributed to the prevalence of G9 rotaviruses in this area.
Adult
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Amino Acid Sequence
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Antigens, Viral
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chemistry
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genetics
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Capsid Proteins
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chemistry
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genetics
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China
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Evolution, Molecular
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Humans
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Infant
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Molecular Sequence Data
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Mutation
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Phylogeny
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Rotavirus
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genetics
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immunology
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physiology
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Serogroup
9.Curative effect of human umbilical cord mesenchymal stem cells for treatment of acute graft-versus-host disease of children after allo-HSCT.
Shu-Min QIAO ; Guang-Hua CHEN ; Yi WANG ; Shao-Yan HU ; Xu-Ding SUN ; De-Pei WU
Journal of Experimental Hematology 2013;21(3):716-720
This study was aimed to investigate the curative effect and safety of human umbilical cord mesenchymal stem cells (hUCMSC) to treat acute graft-versus-host disease (aGVHD) of children after hematopoietic stem cell transplantation (HSCT). HUCMSC were isolated and cultured by collagenase digestion and passage culture. The 3rd to the 5th passage of hUCMSC were used for clinical treatment. Five cases of children acute leukemia achieved complete remission after chemotherapy. Two cases received HLA 3/6 loci matched haploidentical bone marrow HSCT. One case received HLA-matched sibling bone marrow and peripheral blood HSCT. One case received unrelated HLA 4/6 loci matched umbilical cord blood HSCT. One case received unrelated HLA 5/6 loci matched umbilical cord blood HSCT. The children received immunosuppressive therapy after III-IV aGVHD occurring. They received 0.5×10(6)/kg hUCMSC infusion when conventional therapy was ineffective. The results showed that 5 cases of children acute leukemia achieved hematopoietic reconstitution and developed the III-IV grade aGVHD. The five cases of children were infused with hUCMSC. The rash subsided, the liver function was normalized and the gastrointestinal symptoms were improved. The infusion-related adverse reaction did not happen. At present, the 5 children are in remission. It is concluded that allogeneic HSCT is an effective therapeutic method for children with acute leukemia. HUCMSC infusion can be safely and effectively used for the treatment of refractory aGVHD.
Child
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Child, Preschool
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Cord Blood Stem Cell Transplantation
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Female
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Graft vs Host Disease
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therapy
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Hematopoietic Stem Cell Transplantation
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adverse effects
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Humans
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Male
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Mesenchymal Stem Cell Transplantation
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Treatment Outcome
10.Review of subjective well-being scales for elderly
Beibei DAI ; Chunlu BO ; Jianhong QIAO ; Kaiwen DING ; Wei SU ; Cuiping XU
Chinese Journal of Practical Nursing 2017;33(32):2557-2560
Subjective well-being,a comprehensive psychological index for evaluating the quality of life,is closely related to the physical and mental health of the elderly.At present,the mainly Methods for measuring subjective well-being in the elderly is scale. Subjective well-being assessment scales include generic scales and specific scales.Domestic elderly subjective well-being scales commonly imported from abroad.Specific scales for elderly are less in mainland China.This article will review the current research status for subjective well-being assessment scales in the elderly, in order to strengthen the localization research of specific scales and further provides reference for the clinical research of this topic.