1.Tremella Polysaccharides attenuated sepsis through inhibiting abnormal CD4+CD25 high regulatory T cells in mice
Zhenwei SHI ; Yan XU ; Xiaolu LI ; Qingyang LIU
Chinese Journal of Immunology 2016;32(3):313-317
Objective:To determine the effects of TPS on peripheral blood Tregs in sepsis mouse induced by burn plus P.aeruginosa infection.Methods: The experimental mice were separated into five groups randomly ,including sham burn group ,burn plus P.aeruginosa infection group ,burn plus P.aeruginosa infection with TPS (50,100,200 mg/kg) treatment group.Peripheral blood Tregs were isolated with Magnetic Microbeads and cultured in vitro from the day after burn (PBD0) to 4 days after burn(PBD4).IL-10, IFN-γ,IL-4 levels in Tregs culture supernatants were determined by sandwich enzyme-linked immunosorbent assays ( ELISA ) . Purification of CD4+CD25high Tregs and CD4+T cells in C57BL/6 mice were administrated by magnetic beads sorting .Tregs and CD4+T cells were cultured in vitro after joining TPS to without TPS cells as a control .The phenotypes of Tregs were analyzed by flow cytometry , and cytokines were measured by ELISA .Results:Vis-a-vis the results of the untreated group ,TPS could markedly decrease IL-4 and IL-10 secretion level and significantly increase the secretion of IFN-γ,and the secretion of IL-10 level and concentration of TPS dose effect.Vis-a-vis the results of the untreated group ,in vitro experiment ,without stimulation of TPS ,CD4+T cell proliferation and IFN-γwere significantly reduced ( P<0.05 ) and IL-4 levels increased significantly;CD4+T cell proliferation and IFN-γwere significantly increased and IL-4 levels were significantly reduced in the group of TPS with antibody-1;there was no significant difference in CD 4+T cell proliferation and the levels of IFN-γand IL-4 in the group of TPS with antibody-2.Conclusion:TPS could inhibit the abnormal ac-tivities of CD4+CD25highTregs in burn with P.aeruginosa infection mice,at least in part via inhibiting IL-10 secretion,and trigger a shift of Th2 to Th1 with activation of CD4+T cells in burn with P.aeruginosa infection mice.
2.Cloning and analysis of tissue expression of PGC-1αgene in Guangxi Bama mini-pigs
Xueyu YAN ; Qiuwei AO ; Xiaodong YAN ; Qingyang JIANG ; Yafen GUO ; Ganqiu LAN
Acta Laboratorium Animalis Scientia Sinica 2014;(5):27-31
Objective To clone the coding sequence of Guangxi Bama mini-pig PGC-1αgene, and to analyze the expression of PGC-1αgene in various tissues of mini-pigs using RT-PCR and QRT-PCR techniques.Methods The PGC-1αgene coding sequence ( CDS) was amplified by PCR from the cDNA of longissimus muscle of Guangxi Bama mini-pig. The PCR products were inserted into pEASY-T5 vector, transfected E.coli, identified and sequenced.The PGC-1αgene expression in different tissues of the Bama mini-pigs was detected by RT-PCR and QRT-PCR assays.Results The PGC-1αgene CDS of Guangxi Bama mini-pig was cloned.It was 2391 bp in length.It had 99.9%homology with the reference sequence, and had two synonymous mutations that were C-A1105 and G-A1524.The expression level of PGC-1αgene was higher in the heart and kidney, followed by liver, subcutaneous fat and longissimus muscle, but the expression was not de-tected in pancreas of Guangxi Bama mini-pig.Conclusions We have successfully cloned the PGC-1αgene of Guangxi Bama mini-pig, and detected this gene expression in six tissues.The results of this study will provide a basis for studying the effect of PGC-1αon type 2 diabetes mellitus (T2DM) in Bama mini-pigs.
3.Bcl-2 gene-modified neural stem cell transplantation for spinal cord injury in rats
Mei ZHANG ; Yuexin WANG ; Xiaohua HOU ; Jun HONG ; Shengchun YIN ; Yan LI ; Qingyang LIU
Chinese Journal of Comparative Medicine 2016;26(7):35-41
Objective To investigate the bcl?2 gene modification on neurological function recovery in rats with spinal cord injury in neural stem cell transplantation. Methods Cultured rat neural stem cells by Ad?EGFP as vector?mediated side B?cell lymphoma 2 gene ( bcl?2 ) gene transfection of neural stem cells were divided into 3 groups: control group, negative transfection group, bcl?2 transfection group. Use western?blot to detect the expression of bcl?2 protein in neural stem cells before and after transfection. 85 adult female SD rats, successful model 72, were randomly divided into control group, NSCs group, bcl?2?NSCs groups, 24/group, rat acute spinal cord injury model in accordance with a modified Allen’ s method. Assess the motor function by BBB rating and the swash plate test. 7 days after modeling by RT?PCR and Western blot detection of spinal cord injury around HSP27, c?fos gene expression, TUNEL assay apoptosis. Four weeks after model drawn line HE staining and fluorescence microscopy EGFP?labeled NSC survival and distribution of the rats neurophysiological recovery by SEP and MEP. Results bcl?2 gene transfection of rat neural stem cells, bcl?2 transfection group and control group, negative transfection group compared to bcl?2 mRNA and protein levels were expressed ( P < 0. 05 ); lower extremity motor function in rats evaluation of bcl?2?NSCs group than NSCs group, NSCs group than the control group. 72 hours after modeling, bcl?2?NSCs number of apoptotic cells were significantly lower than the control group and NSCs group (P < 0. 05). 7 days after modeling, compared with the control group and NSCs group, bcl?2?NSCs group HSP27 gene and protein expression was significantly higher than that (P < 0. 05), bcl?2?NSCs group c?fos mRNA and protein expression was significantly reduced compared (P < 0. 05). 4 weeks after modeling, HE staining control group showed spinal cord tissue loss and the formation of syringomyelia, no axonal through. NSCs group damage zone few of neuraxis?like structures, syringomyelia smaller, bcl?2?NSCs group showed more nerve axon?like structure, no syringomyelia. EGFP?positive cells labeled:bcl?2?NSCs group the most, NSCs group followed, no control group, and the difference between the groups was statistically significant (P < 0. 05). After the 4th week, SEP and MEP latency period:bcl?2?NSCs group
4.Clinical Study of Modified Shenqi Pill (肾气丸) Plus Tongdu Tiaoshen Acupuncture (通督调神针刺) for Neurogenic Bladder After Spinal Cord Injury of Kidney-Yang Deficiency Syndrome
Meiyan GUO ; Qian FAN ; Yan LI ; Bao YIN ; Sheng JIANG
Journal of Traditional Chinese Medicine 2024;65(2):192-197
ObjectiveTo observe the clinical efficacy of modified Shenqi Pill (肾气丸) plus Tongdu Tiaoshen Acupuncture (通督调神针刺) in the treatment of neurogenic bladder after spinal cord injury of kidney-yang deficiency syndrome. MethodsForty-six patients were randomly divided into 23 cases each in the control group and the treatment group. Both groups were given conventional treatment, i.e. oral methylcobalamin tablets (0.5 mg each time, 3 times a day) and paraplegic conventional acupuncture (once a day, 6 consecutive days a week). The control group was given simple bladder function rehabilitation training on the basis of the conventional treatment; and the treatment group was given modified Shenqi Pill orally (1 dose a day, 150 ml each time, taken warmly in morning and evening) and Tongdu Tiaoshen Acupuncture (once a day, 6 consecutive days per week) in addition to what were given to the control group. The treatment course lasted for 4 weeks. The 24 h urination frequency, 24 h urine leakage frequency, 24 h single urine volume, bladder residual urine volume, international lower urinary tract symptom (LUTS) score, traditional Chinese medicine (TCM) syndrome score were compared between the two groups, and clinical effectiveness and TCM syndrome effectiveness were compared between the two groups after treatment. ResultsTwenty patients in each group were finally analyzed in this study. The number of 24 h urination, the number of 24 h urine leakage, bladder residual urine volume, LUTS score, and the TCM syndrome scores decreased after treatment in both groups, and the 24 h single urine volume increased (P<0.01); and much more improvement was found of each index in the treatment group than in the control group (P<0.05 or P<0.01). The total clinical effectiveness and TCM syndrome effectiveness in the treatment group was 85.00% (17/20) respectively, which were statistically significantly higher than 45.00% (the total clinical effectiveness, 9/20) and 60.00% (TCM syndrome effectiveness, 12/20) in the control group (P<0.01). ConclusionModified Shenqi Pill plus Tongdu Tiaoshen Acupuncture can signi-ficantly improve the clinical symptoms of neurogenic bladder patients after spinal cord injury of kidney-yang deficiency syndrome, having better effectiveness than simple bladder function rehabilitation training, and its mechanism may be related to the improvement of the injured nerve function innervating the bladder.
5.Evaluation on the preliminary classification of carbapenemase in gram-negative rods by the inhibitor enhanced carbapenem inactivation method
Qingyang SUN ; Yan YANG ; Wenjun WEI ; Di LIN ; Jian CHEN ; Xianming ZENG ; Jun CHENG ; Changgui SUN
Chinese Journal of Clinical Laboratory Science 2018;36(1):14-18
Objective To evaluate the reliability of the inhibitor enhanced carbapenem inactivation method (ieCIM) in the detection and preliminary classification of carbapenemase in gram-negative rods.Methods The carbapenem inactivation method (CIM) was modified by adding tazobactam or ethylene diamine tetraacetic acid disodium salt as carbapenemase inhibitors into the reaction system.A total of 198 isolates of Enterobacteriaceae and 35 strains of nonfermenters were collected,and their preliminary classification of carbapenemase was performed by the ieCIM.Meanwhile,their carbapenemase genes were detected by polymerase chain reaction (PCR),and the results were compared with that of the ieCIM.Results Among 198 strains of Enterobacteriaceae,101 were positive for carbapenemase genes,while 99 were detected by the CIM.Among the other 97 strains with negative carbapenemase gene,the results of the ieCIM were also negative.Among 35 strains of nonfermenters,25 were positive for carbapenemase genes,while 24 were detected by the CIM.Among the other 10 strains with negative carbapenemase gene,the results of the CIM were also negative.Using the ieCIM,97.7% (85/87) of strains producing class A carbapenemase and 88.0% (22/25) of strains producing class B carbapenemase were detected.Twelve strains producing class D carbapenemase and 2 strains producing both class A and class B carbapenemase were detected by the ieCIM.The total detection sensitivity and specificity of the ieCIM were 96% and 100%,respectively.Conclusion The ieCIM has the consistent results with the detection method of carbapenemase genes,which may be used to detect and classify carbapenemase in clinical microbiology laboratories.
6.Application progress on thoracic paravertebral nerve block in pediatric perioperative analgesia
Yuying MA ; Yisa SHI ; Yaqin WANG ; Qingyang YAN ; Xuelei JIN
The Journal of Clinical Anesthesiology 2024;40(1):80-84
Thoracic paravertebral nerve block(TPVB)is a regional anesthesia technique that pro-vides ipsilateral somatosensory,motor and sympathetic nerves block segmentally by injecting local anesthetics in the paravertebral space.In recent years,there has been an increasing number of studies on the use of TPVB technique for anesthesia and analgesia in pediatric thoracic and upper abdominal surgery,showing good perioperative analgesic efficacy.This article intends to provide a review of the current applica-tion and progress of TPVB technique for pediatric perioperative analgesia in terms of medication regimens,drug diffusion routes,block methods,clinical application,and complications.
7.Refractory hyponatremia caused by acute intermittent porphyria
Qingyang LI ; Yi REN ; Jingtian HOU ; Yunfeng LIU ; Yan WANG ; Jing YANG
Chinese Journal of Endocrinology and Metabolism 2022;38(9):815-818
Hyponatremia is common in the attack of acute intermittent porphyria(AIP), which can cause epilepsy, coma and other adverse events and endanger the life of patients. Carbohydrate loading therapy is applied to control the attack of AIP in the clinic. But the application of glucose can exacerbate hyponatremia. It is difficult for clinicians to effectively correct hyponatremia while treating AIP with glucose. We reported a case of AIP whose refractory hyponatremia was corrected with short-term low-dose tolvaptan to improve knowledge in management.
8.Causal Association Between Immune Cells and Cervical Cancer: A Two-Sample Mendelian Randomization Study
Jingting LIU ; Yawei ZHOU ; Lingguo KONG ; Qiandan WANG ; Tianxiong SU ; Jianying PEI ; Yan LI
Cancer Research on Prevention and Treatment 2024;51(9):772-778
Objective To investigate potential causative associations between immunophenotype traits and cervical cancer by using two-sample Mendelian randomization (MR) analysis. Methods The genetic instrumental variables (IVs) of 731 immunophenotypes of peripheral blood were obtained from the GWAS Catalog database. The GWAS summary data of cervical cancer were obtained from FinnGen database. The inverse-variance weighted (IVW), weighted mode, weighted median, and MR Egger methods were used for evaluations. The sensitivity analysis and reverse Mendelian randomization analysis were conducted to eliminate bias and reverse causality. The MR Steiger directionality test was further used to ascertain the reverse causal relationship between immune cells and cervical cancer. Results A total of 71 immune cell subtypes associated with cervical cancer were identified, of which 31 had a strong association. The majority of the B cell panel was protective factors for cervical cancer. B-cell activating factor receptor (BAFF-R) was the most frequently expressed molecule in this analysis. It is expressed on several B cell subtypes. The CD20 on IgD+ CD38+ B cell (OR=1.887, 95%CI: 1.078-3.306, P=0.026) is the risk factor for cervical cancer. In cDC panels, the CD123 expression on plasmacytoid dendritic cell (OR=2.48, 95%CI: 1.229-5.003, P=0.011), CD123 expression on CD62L+ plasmacytoid dendritic cell (OR=2.5, 95%CI: 1.231-5.077, P=0.011), CD80 expression on plasmacytoid dendritic cell (OR=2.62,95%CI:1.244-5.515, P=0.011), and CD80 expression on CD62L+ plasmacytoid dendritic cell (OR=2.641, 95%CI: 1.246-5.596, P=0.011) were positively associated with the incidence of cervical cancer. All gynecological cancers in this study have no statistically significant effect on immune cells, according to reverse MR analysis. Conclusion This study emphasized the genetically predicted causality between immune cells and cervical cancer. In clinical practice, it is important to pay attention to the screening of peripheral blood immune cells for patients with cervical cancer.
9.Overview and Analysis of Rare Disease Drugs Approved for Market in China, the United States, the European Union, and Japan in 2023
Shaohong WANG ; Xin LIU ; Qingyang LIU ; Yan TANG ; Bo ZHANG
JOURNAL OF RARE DISEASES 2024;3(2):202-213
To summarize and analyze what the listing in market of orphan drugs in China, the United States, the European Union, and Japan in 2023, and to provide empirical reference to Chinese pharmaceutical enterprises, pharmaceutical management, and new drug research and development departments. Collect the 2023 drug market data released by official drug regulatory agencies in four countries and international organization-China, the United States, the European Union, and Japan categorize the market of rare disease drugs and and their indications, approval dates, R&D status in China, component therapeutic areas, and special review and approval pathways were analyzed. Findings in 2023, 28 orphan drugs were listed in the United States, mostly anti-tumor related, accounting for 32.1% (9 out of 28); 17 orphan drugs were listed in the European Union, anti-tumor related accounting for 47.0% (8 of 17); 22 listed in Japan; and 45 listed in China. In 2023, over 70% orphan drugs listed in the European Union and Japan entered the phase of clinical trials/marketing and applications/approved clinical applications in China (86.4% from Japan and 70.6% from European Union). Currently, there is no qualification accreditation mechanism for orphan drugs in China. However, in 2023, the number of rare disease drugs listed in China was the greatest among the four countries/international organization. It reflects that China′s Catalog for Rare Diseases played a guiding role in drug research and development. Meanwhile, the reform of drug approval and review in China and the issue of rare disease catalogs took place in a short period of time, so the review and approval, as well as research and development of rare disease drugs are still in the catching up stage.
10.Efficacy and safety of Choudongning capsule (CDN)in children with Tourette's syndrome of spleen deficiency and phlegm accumulation.
Na YANG ; Rong MA ; Si-Yuan HU ; Hong LIU ; Hui-Min YAN ; Xi-Xiong XIANG ; Xue-Feng WANG ; Yin-Bo CHEN ; Yu-Yan CHEN ; Fu-Yong JIAO ; Ying DING ; Hao HUANG ; Ling-di WANG
China Journal of Chinese Materia Medica 2016;41(16):3100-3106
To evaluate the efficacy and safety of Choudongning (CDN)capsule in children with Tourette's syndrome of spleen deficiency and phlegm accumulation through a randomized double-blind three-arm controlled phase Ⅲ study in 588 patients from 8 hospitals. The included patients were randomly divided into test group, positive control group and placebo group at the ratio of 3∶1∶1. Patients in the test group orally took CDN capsules and simulated Tiapridal tablets; the patients in positive control group took Tiapridal tablets and simulated CDN capsules; whereas the patients in placebo group orally took the simulated agents of the above two drugs. The treatment course was 6 weeks for three groups. The global grading rates, YGTSS scores and its factor scores, the degree of social function damage, as well as traditional Chinese medicine syndrome efficacy were evaluated as the outcome measures on efficacy. The AEs/ADRs, vital signs and laboratory testing were observed as outcome measures on safety. The total effective rate of YGTSS was 75.92% in the test group, 72.65% in the positive control group, and 37.29% in the placebo group. Non inferiority test stands between the test group and the positive control group, and they were superior to placebo group in efficacy with statistical difference. Significant difference had also been found among the 3 groups in YGTSS tics score, motor tics score, vocal tics, degree of social function damage and traditional Chinese medicine syndrome efficacy. During the study, there were 5 (1.42%)ADRs in the test group, 10 (8.55%)in the positive control group and 3 (2.54%)in the placebo group. The incidence of ADRs in the test group was lower than that in the positive control group, with statistical difference. It is clear to say that CDN capsule can effectively treat the Tourette's syndrome of spleen deficiency and phlegm accumulation. Its efficacy is not inferior to the commonly used Tiapridal tablets, with even less adverse reactions, so it has clinical application value.