scholarly journals Molecular Mechanism of Yisui Shengxue Granule, a Complex Chinese Medicine, on Thalassemia Patients Suffering from Hemolysis and Anemia of Erythrocytes

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Na-Li Chu ◽  
Zhi-kui Wu ◽  
Xin-Hua Zhang ◽  
Su-Ping Fang ◽  
Wen-Juan Wang ◽  
...  
Keyword(s):  
Chinese Medicine ◽  
Mrna Expression ◽  
Expression Level ◽  
Blood Parameter ◽  
Rt Pcr ◽  
Relative Expression Level ◽  
Positive Clinical Effect ◽  
Increasing Trend ◽  
Yisui Shengxue Granule ◽  
Relative Mrna Expression

The objective of this study was to investigate the therapeutic biological mechanism of Yisui Shengxue Granule (YSSXG), a complex Chinese medicine, on the hemolysis and anemia of erythrocytes from patient with thalassemia disease. Sixteen patients with thalassemia (8 cases ofα-thalassemia and 8 cases ofβ-thalassemia) disease were collected and treated with YSSXG for 3 months. The improvements of blood parameter demonstrated that YSSXG had a positive clinical effect on patients with thalassemia disease. For patients withα-thalassemia disease, RT-PCR showed that YSSXG upregulated the relative mRNA expression level ofα-globin toβ-globin and downregulated DNMT1, DNMT3a, and DNMT3b mRNA compared with pretreatment. Western blotting showed that YSSXG downregulated the expression of DNMT1 and DNMT3a. For patients withβ-thalassemia disease, the relative expression level ofAγ-globin toα-globin had an increasing trend and the level of BCL11A mRNA expression obviously increased. For all patients, RT-PCR showed that YSSXG upregulated mRNA expression of SPTA1 and SPTB. Activities of SOD and GSH-Px significantly increased and MDA obviously reduced on erythrocyte and blood serum after YSSXG treatment. TEM showed that YSSXG decreased the content of inclusion bodies. Activities of Na+K+-ATPtase and T-ATPtase of erythrocyte increased significantly after YSSXG treatment. This study provides the basis for mechanisms of YSSXG on thalassemia suffering with hemolysis and anemia of erythrocytes from patient.

scholarly journals Cytokine and Chemokine mRNA Expressions after Mycobacterium tuberculosis-Specific Antigen Stimulation in Whole Blood from Hemodialysis Patients with Latent Tuberculosis Infection

Diagnostics ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 595
Author(s):  
Ji Young Park ◽  
Sung-Bae Park ◽  
Heechul Park ◽  
Jungho Kim ◽  
Ye Na Kim ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mrna Expression ◽  
Whole Blood ◽  
Latent Tuberculosis Infection ◽  
Latent Tuberculosis ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Healthy Individuals ◽  
Relative Mrna Expression ◽  
Mrna Expression Levels

There have been few reports on the kinetics of hemodialyzed (HD) patients’ immune responses in latent tuberculosis infection (LTBI). Therefore, in the present study, messenger ribonucleic acid (mRNA) expression levels of nine immune markers were analyzed to discriminate between HD patients with LTBI and healthy individuals. Nine cytokines and chemokines were screened through relative mRNA expression levels in whole blood samples after stimulation with Mycobacterium tuberculosis (MTB)-specific antigens from HD patients with LTBI (HD/LTBI), HD patients without LTBI, and healthy individuals, and results were compared with the QuantiFERON-TB Gold In-Tube (QFT-GIT) test. We confirmed that the C-C motif chemokine 11 (CCL11) mRNA expression level of the HD/LTBI group was significantly higher than the other two groups. Especially, the CCL11 mRNA expression level of the >0.7 IU/mL group in the QFT-GIT test was significantly higher than the <0.2 IU/mL group in the QFT-GIT test and the 0.2–0.7 IU/mL group in the QFT-GIT test (p = 0.0043). The present study reveals that the relative mRNA expression of CCL11 was statistically different in LTBI based on the current cut-off value (i.e., ≥0.35 IU/mL) and in the >0.7 IU/mL group. These results suggest that CCL11 mRNA expression might be an alternative biomarker for LTBI diagnosis in HD patients.

Gene expressions of TS and MDR-1 are predictive factors for chemosensitivity and survival in esophageal cancer with fluoropyrimidine-based chemotherapy

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 14120-14120
Author(s):  
K. Uchida ◽  
K. Hayashi ◽  
K. Kudo ◽  
H. Kuramochi ◽  
M. Yamamoto
Keyword(s):  
Esophageal Cancer ◽  
Mrna Expression ◽  
Overall Response Rate ◽  
Response Rate ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Gene Expressions ◽  
Polymerase Chain ◽  
Relative Mrna Expression ◽  
Mdr 1

14120 Background: Fluoropyrimidines are widely used in chemotherapy regimens for esophageal cancer. To test the hypotheses of whether the relative mRNA expression of the enzyme TS, DPD and MDR-1 were associated with response to in esophageal cancer. We investigated the associations reported between intratumoral TS, DPD and MDR-1 gene expressions and the response with 5-FU based chemotherapy for patients with esophageal cancer. Methods: Twenty six pts with esophageal cancer were treated with 5-FU based chemotherapy (14 pts were treated with chemotherapy only, and 12 pts were chemoradiotherapy). mRNA was isolated from frozen tumor specimens, and relative expression levels of each gene/β-actin were measured using a quantitative reverse transcription polymerase chain reaction (RT-PCR) (Taqman®) system. Results: The overall response rate was 50.0%. Each mRNA expression was detectable in 26 patients. TS expressions were significantly lower in the responding tumors compared to the non-responders respectively (P=0.009). There were no significant associations between MDR-1 expression and the response. By setting up a cut-off level for TS and MDR-1 of Median, combining the two gene expression two-dimensionally revealed a relationship with the response (P=0.037). Additionally, MDR-1 expressions were statistically significant predictors of prolonged survival (P=0.0084). Conclusions: These results suggest that intratumoral TS and MDR-1 expressions are prognostic factors for survival after 5-FU based chemotherapy in esophageal cancer. No significant financial relationships to disclose.

Novel alternative splice variant of IGF-1R and its mRNA expression patterns in BaMa and Landrace pigs

2019 ◽  
Author(s):  
Rui Yang ◽  
Lijie Dong ◽  
Songcai Liu ◽  
Yunyun Cheng ◽  
Wenzhen Wei ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Splice Variant ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Expression Level ◽  
Alternative Splice Variant ◽  
Nucleotide Polymorphism ◽  
Rt Pcr ◽  
Single Nucleotide ◽  
Muscle Tissues

The transcript variants of Insulin-like growth factor 1 receptor (IGF-1R) and their expression profiles had never been illuminated in pigs until now. Herein, we identified IGF-1R AS02 as a novel splice variant of IGF-1R gene by RT-PCR and analyzed its mRNA expression level by qRT-PCR in liver, cartilage and muscle tissues, while also detecting the single-nucleotide polymorphism (SNP) site near the splice site of the IGF-1R gene (in intron 19) of BaMa and Landrace pigs. Results demonstrated that the IGF-1R AS02 variant showed a significantly (P less than 0.05) higher expression level in cartilage than in muscle and liver across two pig breeds respectively. The expression level of the normal transcript (IGF-1R ISO01) of IGF-1R in cartilage was markedly lower than that in the other two tissues (P less than 0.05). In cartilage, IGF-1R ISO01 expression was higher in BaMa than in Landrace (P less than 0.05), while the expression level of IGF-1R AS02 was lower in BaMa than in Landrace (P less than 0.05). The SNP was detected in intron 19 of the IGF-1R gene of BaMa and Landrace pigs. These results contributed to facilitating a better understanding of IGF-1R gene in pigs.

The Feature of CD3-zeta Chain Gene Expression in Mononuclear Cells without and with Stimulation by Different Factors from Umbilical Cord Blood.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3870-3870
Author(s):  
Shaohua Chen ◽  
Yangqiu Li ◽  
Lijian Yang ◽  
Si Chen
Keyword(s):  
Gene Expression ◽  
T Cells ◽  
Cord Blood ◽  
Mrna Expression ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Healthy Adults ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Relative Mrna Expression

Abstract CD3 zeta gene is required for efficient TCR expression and plays a central role in the signal-transducing events leading to T and NK-cell activation and proliferation. Umbilical cord blood (UCB) has been used successfully as a source of allogeneic transplantation and UCB T cells have showed capacity for production the specific CTL by tumor associated antigen ex vivo. In order to investigate the feature of CD3 zeta gene expression pattern in cord blood, UCB T cell without or with stimulation by different stimulators, including PHA, IL-2, CD3 monoclonal antibody (McAb), CD28 McAb + IL-2 and PML-RARα peptide) were used to analyze. By using Real-Time PCR with SYBR Green I technique, the expression level of CD3 zeta gene was analyzed in T-cells from 60 cases of UCB before and after T-cells culture at different time points (5–20days), β2-microglobulin gene was used as an endogenous reference. The relative mRNA expression level of CD3 zeta gene was used by the 2- ΔCt method. According to melting curve, polymorphism of nucleotide sequence was determined by PCR products direct sequencing. 60 cases healthy adults served as controls. The results showed that CD3 zeta gene was expressed in all cases from both UCB and healthy adults. The mean value 6.7%±5.56% of relative mRNA expression level of CD3 zeta gene was found in 60 UCB cases, the expression level under 1.0% was detected in 4 cases, over 10% in 14 cases and a high expression of 25.53% only in one case. In contrast, 3.1%±2.23% of relative mRNA expression level of CD3 zeta gene was detected in 60 cases healthy adults. The expression of the CD3 ζ gene from the healthy adults is more concentrated and the highest expression is only 9.34%. Compare with the healthy adults, a significant higher expression of CD3 zeta gene was found from UCB (P=0.000). Polymorphism and mutation of CD3 zeta gene were not identified by sequence analysis in both UCB and healthy samples. For the culture cells, CD3 zeta gene expression level in initial culture (5–10days) was increased after different stimulation. A higher expression lever was found in combined CD3 MCAb + CD28 McAb with or without PML-RARα peptide than in stimulation with PHA or IL-2 alone. The CD3 zeta gene expression lever in UCB T cells induced by combined PML-RARα peptide was 6.37 times as much as unstimulated cells at 10 days, whereas the CD3 zeta gene expression lever in UCB T cells stimulated by IL-2, CD3 McAb plus CD28 McAb was 5.83 times higher than that from un-stimulated cells. However, when the duration of T cells culture was prolonged, the expression of CD3 zeta was gradually reduced in all groups after 10 days. In conclusion, this is to our knowledge, the first description of CD3 zeta gene expression in UCB. Our data suggest that a higher expression of CD3 zeta gene could be found in UCB than in adult peripheral blood, and up-regulation of CD3 zeta gene can also be achieved after stimulation with PHA, IL-2, CD3 McAb + CD28 McAb + IL-2 with or without PML-RARα peptide, respectively. In addition, combined CD3 McAb + CD28 McAb with/without PML-RARα peptide showed a significantly higher capacity for promoting proliferation. The down-expression of CD3 zeta in cultured T cells after 10 days remains an open question.

scholarly journals Role of Annexin A2 isoform 2 on the aggregative growth of dermal papillae cells

2018 ◽  
Vol 38 (6) ◽  
Author(s):  
Jing Gu ◽  
Yinni Ma ◽  
Lijia Yang ◽  
Feng Wang ◽  
Cao Lei ◽  
...  
Keyword(s):  
Western Blot ◽  
Mrna Expression ◽  
Expression Vector ◽  
Dermal Papilla ◽  
Annexin A2 ◽  
Expression Level ◽  
Rt Pcr ◽  
Aggregative Growth ◽  
Dermal Papillae

The dermal papilla is a major component of hair, which signals the follicular epithelial cells to prolong the hair growth process. Human Annexin A2 was preliminarily identified by two-dimensional gel electrophoresis (2-DE), MALDI-TOF-MS and database searching. The aim of the present study was to explore the role of Annexin A2 in the aggregative growth of dermal papillae cells (DPC). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were adopted to detect the expression of Annexin A2. And siRNA technique was used to suppress the expression of Annexin A2. Construction of over-expression vector was used to up-regulate the expression of Annexin A2. Cell Counting Kit 8 (CCK-8) and proliferating cell nuclear antigen (PCNA) were taken to detect the proliferation of DPC. The expression of Annexin A2 mRNA was up-regulated in passage 3 DPC compared with passage 10 DPC by RT-PCR. In line with the results at the mRNA level, Western blot analysis revealed that Annexin A2 isoform 2 was up-regulated significantly in passage 3 DPC compared with passage 10 DPC. The Annexin A2 isoform 2 siRNA was synthesized and transfected into passage 3 DPC. RT-PCR data showed the mRNA expression of Annexin A2 isoform 2 was suppressed in passage 3 DPC. Western blot results showed the expression level of Annexin A2 isoform 2 and PCNA were suppressed in passage 3 DPC. CCK-8 results showed that the proliferation of passage 3 DPC was suppressed (P < 0.05). Recombinant plasmid PLJM-Annexin A2 isoform 2-expression vector were constructed and were transfected into passage 10 DPC. RT-PCR data showed the mRNA expression of Annexin A2 isoform 2 was up-regulated in passage 10 DPC. Western blot results showed the expression level of annexin A2 isoform 2 and PCNA were up-regulated in passage 10 DPC. CCK-8 assay showed the proliferation of DPC was stimulated compared with control group (*P < 0.05). Our study proved that Annexin A2 isoform 2 may participate in regulating the proliferation of DPC and may be related to aggregative growth of dermal papilla cells. Therefore, our study suggests that Annexin A2 may be linked to hair follicle growth cycle.

Synergic Effects of Immunosuppressive Agents on T Lymphocyte Function and Cytokine Expression Pattern.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5213-5213
Author(s):  
Mao Fang Lin ◽  
Yan Ling Jiang
Keyword(s):  
Protein Expression ◽  
Mrna Expression ◽  
T Lymphocyte ◽  
Immunosuppressive Agents ◽  
Reaction System ◽  
Expression Level ◽  
Lymphocyte Function ◽  
Relative Expression ◽  
Relative Expression Level ◽  
Ifn Γ

Abstract The regimen including mycophenolate mofetil (MMF) and cyclosporine A (CsA) has been successfully used as an effective method in the prophylaxis of acute graft versus host disease. Herewith we investigated the effect of CsA, mycophenolic acid (MPA) alone or in combination with anti B7–1 mAb on proliferation of T lymphocytes and their possible mechanisms. The T lymphocyte reaction system was established in vitro and was detected by 5-bromo-2-deoxyuridine incorporation method; the expression of IL-2, IFN-γ and IL-10 on mRNA and protein level were detected by RT-PCR and ELISA respectively. Experimental concentration of MPA (50μmol/L) and CsA(0.33μmol/L) inhibited the T lymphocyte proliferation significantly. CsA in combination with anti B7–1 mAb(10mg/L) had a much stronger effect on the inhibition(p<0.01). There was no difference between groups treated either with MPA or with the combination of MPA and anti B7–1 mAb. Both of CsA and MPA could inhibit the protein expression of IL-2 and IFN-γ. The protein expression of IL-2 were 99.70±9.15 pg/ml, 48.19±8.67pg/ml and 42.73±14.64 pg/ml in groups of control, treated with CsA or MPA respectively. The protein expression of IFN-γ in the group treated with CsA or MPA decreased significantly than the control (6.81±5.24 pg/ml, 7.87±4.22 pg/ml vs 82.42±25.55 pg/ml, p<0.05). Compared with that treated with CsA alone, The protein expression of IFN-γ in the group treated with CsA in combination with anti B7–1 mAb decreased significantly (0.30±0.52 pg/ml vs 6.81±5.24 pg/ml, p<0.05). Both of CsA and MPA increased the protein expression of IL-10. Compared with MPA alone, MPA in combination with anti B7–1 mAb increased significantly the expression of IL-10 (941.90±56.61 pg/ml vs 770.95±126.85 pg/ml, p<0.05). The relative expression level of IL-2 mRNA (IL-2/GAPDH) and IFN-γ mRNA (IFN-γ /GAPDH) in the CsA or MPA group was significantly lower than the control (p<0.01). The relative expression level of IFN-γ mRNA in combination group of CsA with anti B7–1 mAb decreased even more significantly (0.38±0.05 vs 0.59±0.02, p<0.01). On the other hand, this combination could up-regulate IL-10 mRNA expression (1.80±0.13 vs 1.38±0.06, p<0.01). Combination of MPA with anti-B7–1 mAb showed a statistically significant increase in IL-10 mRNA expression (1.28±0.06 vs 0.84±0.09, p<0.01) as compared with MPA alone. In conclusion, MPA and CsA induced the changes of cytokine expressive spectrum and the Th1 and Th2 shift might be involved in the immunosuppressive effect. The combination of immunosuppressive agents, especially with anti B7–1 mAb had a synergic effect.

scholarly journals Evaluation of the Correlation between the mRNA Expression Levels of ystA and ymoA Genes in Y. enterocolitica Strains with Different Enterotoxic Properties

Pathogens ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1136
Author(s):  
Agata Bancerz-Kisiel ◽  
Karolina Lipczyńska-Ilczuk
Keyword(s):  
Mrna Expression ◽  
Clinical Symptoms ◽  
Expression Level ◽  
Common Source ◽  
Expression Levels ◽  
Relative Expression ◽  
Source Of Infection ◽  
Relative Expression Level ◽  
Causative Agents ◽  
Mrna Expression Levels

Yersinia enterocolitica is one of the main causative agents of human diarrhea. Pigs are a reservoir and the most common source of infection for humans. The aim of this study was to analyze the expression of ystA and ymoA genes in Y. enterocolitica strains with different enterotoxic properties, isolated from humans and pigs. The experiment involved two groups of Y. enterocolitica strains producing and not producing enterotoxin YstA, which were isolated from humans and pigs. All strains were ystA- and ymoA-positive. The expression of ystA and ymoA genes was analyzed by quantitative real-time PCR (qPCR). The relative expression level of the ystA gene was significantly higher than the expression level of the ymoA gene in Y. enterocolitica strains isolated from humans with clinical symptoms of yersiniosis. In other strains, a significant decrease in ystA gene transcription was observed, and the relative expression level of the ymoA gene was significantly higher than the expression level of the ystA gene. Statistically significant differences were not observed in either group of strains isolated from pigs. The results of our study revealed a correlation between mRNA expression levels of ystA and ymoA genes in Y. enterocolitica strains isolated from humans.

5′-Nucleosidase and Deoxycytidine Kinase Gene Expressions in High-Risk Myelodysplastic Syndrome (MDS).

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3434-3434
Author(s):  
Keijiro Suzuki ◽  
Takeshi Sugawara ◽  
Yoji Ishida
Keyword(s):  
Overall Survival ◽  
High Risk ◽  
Clinical Outcome ◽  
Mrna Expression ◽  
Expression Level ◽  
Deoxycytidine Kinase ◽  
Rt Pcr ◽  
Significant Difference ◽  
The Mean ◽  
Classification Quantitative

Abstract Various chemotherapeutic regimens including cytarabine (ara-C) for MDS patients have been less effective than those for AML patients. Althogh alterations in the expression of genes involved in ara-C metabolism have been reported to be related to the clinical outcome in AML patients, there is no report about MDS. Deoxycytidine kinase (dCK) is the rate-limiting enzyme in this activation process being the first phosphorylation of ara-C. Cytoplasmic 5′-nucleotidase (5′-NT) dephosphorylates ara-CMP. The concentration of the intracellular ara-CTP, active form of ara-C, depends on the activities of two enzymes. To determine whether the level of expression of 5′-NT and dCK is implicated in clinical outcome in patients with high-risk MDS (RAEB/RAEB-t), we analyzed the mRNA expression of these at diagnosis in bone marrow (BM) cells of patients with RAEB/RAEB-t using real-time PCR (rt-PCR). Materials and Methods: BM cells obtained from 22 patients (male: female=16:6) with untreated high-risk MDS. The mean age of all patients was 66.4±7.9 y. 7 patients (31.8%) had RAEB-t subtype and 15 (68.2%) patients RAEB subtype of the FAB classification. Quantitative rt-PCR of 5′-NT or dCK mRNA of BM cells in patients and 12 healthy volunteers was performed by LightCycler using β-actin as housekeeping control. For each sample, a ratio of 5′-NT or dCK value/β-actin value was calculated and considered as the level of 5′-NT or dCK mRNA expression. Results: 15 of patients received ara-C containing regimen (7 patients had supported care only). Complete remission was obtained in 7 patients. Other 8 patients were chemoresistant. The median overall survival (OS) of all patients was 22.9 months (range 2.0–91.0), and that of patients with chemotherapy was 20.0 (7–91). The median post-chemotherapy survival (PCS) was 15.0 months (5.0–43.0). At diagnosis, the mean and median expression level of 5′-NT mRNA in all patients was 1.56 (SD 1.61) and 1.36 (range 0.77–6.77), respectively (control: mean 0.23, SD 0.06 and median 0.67, range: 0.17–0.34, p&lt;0.01). The expression of dCK mRNA did not show any significant difference between patients and control. Patients with chemotherapy whose BM cells were higher level of 5′-NT expression (greater than mean value) had shorter OS (14.0 months vs. 26.0 months, p&lt;0.01) and shorter PCS (10.0 months vs. 17.0 months, p=0.011). Figure Figure Conclusion: These data suggest that the expression level of 5′-NT mRNA might be a candidate for prognostic factor in RAEB/RAEB-t. Patients with an increase of 5′-NT mRNA at diagnosis show shorter overall survival and chemoresistance.

scholarly journals Cytokine and Chemokine mRNA Expressions After Mycobacterium Tuberculosis-Specific Antigen Stimulation in Whole Blood From Hemodialysis Patients with Latent Tuberculosis Infection

2020 ◽  
Author(s):  
Ji Young Park ◽  
Sung-Bae Park ◽  
Heechul Park ◽  
Jungho Kim ◽  
Ye-Na Kim ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mrna Expression ◽  
Whole Blood ◽  
Latent Tuberculosis Infection ◽  
Specific Antigen ◽  
Latent Tuberculosis ◽  
Tuberculosis Infection ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Relative Mrna Expression

Abstract There have been few reports on the kinetics of hemodialyzed (HD) patients’ immune responses in latent tuberculosis infection (LTBI). Nine cytokines and chemokines were screened through relative mRNA expression levels in whole blood samples after stimulation with Mycobacterium tuberculosis (MTB)-specific antigens from HD patients with LTBI (HD/LTBI), HD patients without LTBI, and healthy individuals and results were compared with QuantiFERON-TB Gold In-Tube (QFT-GIT) test.Only the CCL11 mRNA expression level of the HD/LTBI group was significantly higher than the other two groups (P = 0.028). CCL11 mRNA expression level of the > 0.7 IU/mL group in QFT-GIT test was significantly higher than the < 0.2 IU/mL group in QFT-GIT test and the 0.2-0.7 IU/mL group in QFT-GIT test (P = 0.0043). The present study reveals that the relative mRNA expression of CCL11 was statistically different in LTBI based on the current cut-off value (i.e., ≥ 0.35 IU/mL) and in the > 0.7 IU/mL group. These results suggest that CCL11 might be an alternative biomarker for LTBI diagnosis in HD patients.

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