1.IL-10 enhances promoter activity of ILT4 gene and up-regulates its expression in THP-1 cells.
Xiaoli, XU ; Ping, ZOU ; Lijuan, CHEN ; Guannan, JIN ; Hao, ZHOU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(5):594-8
This study examined the effect of IL-10 on immunoglobulin-like transcript (ILT4) expression of human monocytic leukemic cell line THP-1, especially the role of the ILT4 promoter activity. ILT4 promoter area was amplified by PCR, and was cloned into the eukaryotic expressing vector pGL3-Basic. The pGL3-ILTP obtained was tested by double endonuclease digestion and sequencing. Then, the recombinant plasmid was transfected into THP-1 cells by using lipofectamine. After culture with IL-10 for 12 h, the mRNA extracted from THP-1 cells was detected by RT-PCR and the protein was detected by FACS. The dual-luciferase reporter assay system was employed to detect the activity of ILT4 promoter with or without IL-10. The results showed that the activity of pGL3-ILTP was significantly increased and was more than ten times that of pGL3-Basic cells. After culture with IL-10 for 12 h, the expression of ILT4 protein and its mean fluorescence intensity (MFI) were increased. Moreover, the mRNA was remarkably higher than that of the control group. Dual-luciferase reporter assay revealed that ILT4 promoter was much more activated after being treated with IL-10. We were led to conclude that pGL3-ILTP containing ILT4 promoter was constructed successfully. The expression of ILT4 could be up-regulated by IL-10 both at the transcriptional and translational level. Furthermore, ILT4 promoter could be much more active after addition of IL-10. This study suggests that IL-10 up-regulates ILT4 expression on monocytes via increasing ILT4 gene promoter activity, which may have implication for inducing transplantation tolerance in clinical practice.
2.The impact of melatonin on inflammatory response after acute spinal cord injury
Yusheng XU ; Xingchen LI ; Weilin JIN ; Peisong WANG ; Guannan ZENG ; Song ZHANG ; Hao CUI
Chinese Journal of Physical Medicine and Rehabilitation 2014;36(4):260-265
Objective To explore the effects of melatonin (MT) on the expression of interleukin (IL)-10,interleukin-6 and interleukin-8 as well as the inflammatory reaction and nerve repair after acute spinal cord injury (SCI).Methods One hundred and eight Sprague-Dawley rats were randomly divided into a spinal cord injury group (group A),an MT treatment group (group B) and a sham operation group (group C),each with 36 rats.SCI models were established in the rats of groups A and B using a version of Allen's weight drop method (50gcf at the T12 level).Group C had removal of the lamina only.Ten minutes later,group A was injected with 5% ethanol in saline (the MT solvent) and group B with 100 mg/kg of melatonin preparation.At 6,12,18 and 24 hours,IL-6,IL-8 and IL-10 levels in serum were detected in 6 rats of each group.At 18 hours post-surgery,spinal cord specimens were taken from 6 rats of each group for hematoxylin eosin staining,morphological examination and immunohistochemical SP detection of IL-10 expression.Results The specimens of group A showed inflammatory reaction and ulceration at 48 h; groups B and C had no ulcers.Group B showed the highest levels of IL-10 in serum and IL-10 mRNA in the spinal cord,while group C showed the lowest level.The differences were statistically significant.Group A had the highest levels of IL-6 and IL-8 and group C had the lowest.The difference between group B and groups A and C was significant.The morpho-logical observation showed that after melatonin treatment the IL-10 levels in the spinal cord's central canal and around the gray matter improved.Conclusions Melatonin can improve nerve lipid peroxidation and inflammatory reaction in the treatment of spinal cord injury by increasing IL-10 expression and inhibiting IL-6 and IL-8 expression.
3.IL-10 Enhances Promoter Activity of ILT4 Gene and Up-regulates Its Expression in THP-1 Cells
XU XIAOLI ; ZOU PING ; CHEN LIJUAN ; JIN GUANNAN ; ZHOU HAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(5):594-598
This study examined the effect of IL-10 on immunoglobulin-like transcript (ILT4) expression of human monocytic leukemic cell line THP-1, especially the role of the ILT4 promoter activity. ILT4 promoter area was amplified by PCR, and was cloned into the eukaryotic expressing vector pGL3-Basic. The pGL3-ILTP obtained was tested by double endonuclease digestion and sequencing. Then, the recombinant plasmid was transfected into THP-1 cells by using lipofectamine. After culture with IL-10 for 12 h, the mRNA extracted from THP-1 cells was detected by RT-PCR and the protein was detected by FACS. The dual-luciferase reporter assay system was employed to detect the activity of ILT4 promoter with or without IL-10. The results showed that the activity of pGL3-ILTP was significantly increased and was more than ten times that ofpGL3-Basic cells. After culture with IL-10 for 12 h, the expression of ILT4 protein and its mean fluorescence intensity (MFI) were increased. Moreover, the mRNA was remarkably higher than that of the control group. Dual-luciferase reporter assay revealed that ILT4 promoter was much more activated after being treated with IL-10.We were led to conclude that pGL3-ILTP containing ILT4 promoter was constructed successfully.The expression of ILT4 could be up-regulated by IL-10 both at the transcriptional and translational level. Furthermore, ILT4 promoter could be much more active after addition of IL-10. This study suggests that IL-10 up-regulates ILT4 expression on monocytes via increasing ILT4 gene promoter activity, which may have implication for inducing transplantation tolerance in clinical practice.
4.Acupuncture with Manipulation for Lumbar Disc Herniation in Remission of 50 Cases:A Randomized Controlled Trial
Chun CHEN ; Jiao JIN ; Jingxuan MO ; Hai LIN ; Fudong SHI ; Guojun WANG ; Guannan WU ; Shimin ZHANG
Journal of Traditional Chinese Medicine 2024;65(10):1026-1032
ObjectiveTo evaluate the clinical effectiveness and safety of acupuncture with manipulation for lumbar disc herniation in remission period. MethodsOne hundred and four patients with lumbar disc herniation in remission were randomly divided into a treatment group and a control group, with 52 cases in each group. Treatment group applied acupuncture with manipulation of pointing, pulling, and shaking. Acupoints were selected as lumbar Jiaji (EX-B2, bilateral), Ashi point, Shenshu (BL 23, bilateral), Huantiao (GB 30, bilateral), Weizhong (BL 40, opposite side of the affected area), Chengshan (BL 57, opposite side of the affected area). The control group applied lumbar traction plus acupoint ultrasonic pulse penetration therapy (acupoints selection same as the treatment group); 20 minutes each time, 3 times a week, a total of 3 weeks for both groups. The primary outcome was the improvement rate of lumbar disc herniation symptoms and signs, which was calculated at 1 week of treatment, 3 weeks of treatment, 1 month follow-up, and 3 months follow-up, respectively; the secondary outcome were the Japanese Orthopaedic Association (JOA) scores, Visual Analogue Scale (VAS) scores, and Oswestry Disability Index (ODI) scores (including ODI total scores, sitting scores and standing scores), which were evaluated before treatment, 1 week of treatment, 3 weeks of treatment, 1 month follow-up, and 3 months follow-up; clinical effectiveness was assessed at 3 months follow-up; and the occurrence of adverse events in the participants, as well as blood routine, urine routine, stool routine, and electrocardiograms before and after the treatment were recorded to evaluate safety. ResultsTwo patients from each group fell out, and 50 patients of each group were included in the outcome analysis ultimately. The scores of lumbar disc herniation symptoms and signs improved more in the treatment group than in the control group at 1 week of treatment, 3 weeks of treatment, 1 month follow-up and 3 months follow-up (P<0.01). The JOA scores of participants in both groups at 1 week of treatment, 3 weeks of treatment, 1 month follow-up and 3 months follow-up were higher than those before treatment in the same group, and the VAS scores, ODI total scores, ODI sitting scores and standing scores were significantly lower than those before treatment in the same group (P<0.05), and the JOA scores of patients in the treatment group were higher than those of the control group at all time points, and the VAS scores, ODI total scores, ODI sitting scores and standing score were lower than those of the control group (P<0.05). At the 3 months follow-up, the excellent rate of the treatment group was 70.00% (35/50) better than that of the control group, which was 50.00% (25/50) (P<0.05). There were no abnormalities in blood, urine, stool routines and electrocardiograms before and after treatment in both groups, and no adverse events occurred. ConclusionAcupuncture with manipulation of pointing, pulling, and shaking for treating patients with lumbar disc herniation in remission has a better safety on pain relief and improving quality of life, and the effectiveness is better than lumbar traction plus acupoint ultrasonic pulse penetration therapy.
5.Clinical characteristics and risk factors for mortality in cancer patients with COVID-19.
Junnan LIANG ; Guannan JIN ; Tongtong LIU ; Jingyuan WEN ; Ganxun LI ; Lin CHEN ; Wei WANG ; Yuwei WANG ; Wei LIAO ; Jia SONG ; Zeyang DING ; Xiao-Ping CHEN ; Bixiang ZHANG
Frontiers of Medicine 2021;15(2):264-274
Patients with cancer are at increased risk of severe infections. From a cohort including 3060 patients with confirmed COVID-19, 109 (3.4%) cancer patients were included in this study. Among them, 23 (21.1%) patients died in the hospital. Cancer patients, especially those with hematological malignancies (41.6%), urinary carcinoma (35.7%), malignancies of the digestive system (33.3%), gynecological malignancies (20%), and lung cancer (14.3%), had a much higher mortality than patients without cancer. A total of 19 (17.4%) cancer patients were infected in the hospital. The clinical characteristics of deceased cancer patients were compared with those of recovered cancer patients. Multivariate Cox regression analysis indicated that a Nutritional Risk Screening (NRS2002) score ⩾ 3 (adjusted hazard ratio (HR) 11.00; 95% confidence interval (CI) 4.60-26.32; P < 0.001), high-risk type (adjusted HR 18.81; 95% CI 4.21-83.93; P < 0.001), tumor stage IV (adjusted HR 4.26; 95% CI 2.34-7.75; P < 0.001), and recent adjuvant therapy (< 1 month) (adjusted HR 3.16; 95% CI 1.75-5.70; P < 0.01) were independent risk factors for in-hospital death after adjusting for age, comorbidities, D-dimer, and lymphocyte count. In conclusion, cancer patients showed a higher risk of COVID-19 infection with a poorer prognosis than patients without cancer. Cancer patients with high-risk tumor, NRS2002 score ⩾ 3, advanced tumor stage, and recent adjuvant therapy (< 1 month) may have high risk of mortality.
COVID-19
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Hospital Mortality
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Neoplasms
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Prognosis
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Retrospective Studies
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Risk Factors
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SARS-CoV-2