1.Packaging and identification of miR-137 overexpression lentivirus
Dong LYU ; Chunmei LIANG ; Mingying LI ; Jingwen YIN ; Xudong LUO ; Juda LIN ; Guoda MA
Journal of Jilin University(Medicine Edition) 2017;43(4):694-697,前插1
Objective:To construct lentiviral vector which can overexpression miR-137 and produce lentivirus by lentivirus packaging system, and to explore its infection efficiency and expression in HEK293T cells.Methods: miR-137 sequence was chemically synthesized and cloned into lentiviral vector GV209, and the recombinant plasmid containing human miR-137 was obtained and identified.Then miR-137 recombinant plasmid together with two helper plasmids were transfected into HEK293T cells using Lipofectamine 2000.After the HEK293T cells were infected in multiplicity of infection(MOI) 40 for 48 h, the expression of green fluorescent protein (GFP) was observed by fluorescence microscope and the expression level of miR-137 was detected by fluorescence quantitative PCR.Results:The sequencing results showed that the inserted gene sequence was completely consistent with the published human miR-137 gene sequence in GenBank.The GFP was observed in the HEK293T cells infected with miR-137 overexpression lentivirus under fluorescence microscope.The fluorescence quantitative PCR results showed that the expression level of miR-137 in the cells infected with overexpression lentivirus was 12.74 times higher than that in the control cells.Conclusion:The lentivirus containing miR-137 gene is successful packaged, and it could efficiently infect the HEK293T cells.
2.Characteristics of liquid food distribution in the stomach of 52 patients with postprandial distress syndrome-predominated functional dyspepsia
Lin LYU ; Jing WANG ; Xudong TANG ; Suiping HUANG ; Fengyun WANG ; Nan KANG ; Ting CHEN ; Enlin ZHU ; Xiaoge WANG
Chinese Journal of Digestion 2015;(7):455-459
Objective To explore the distribution of the liquid food in the stomach of patients with postprandial distress syndrome (PDS)-predominated functional dyspepsia (FD)and the mechanism of its gastric dynamic dysfunction.Methods Fifty-two patients with PDS and 18 healthy volunteers underwent gastric emptying examination with ultrasound scanner provided.The proximal and distal gastric area and volume,area and volume ratio of proximal and distal gastric as well as emptying rate of proximal and distal gastric were calculated at fasting,maximum satiety,30 min,60 min,90 min and 120 min after meal. Student′s t test andχ2 test were performed for statistical analysis.Results Proximal gastric volume of PDS group at the time of fasting and 120 min after meal were (9.06±7.88)and (24.72±24.02)cm3 ,which were larger than those of healthy control group ((5 .96 ± 2.13 )cm3 ,(19.48 ± 12.32 )cm3 ),and the differences were statistically significant (t=-1 .637 and -0.875 ,both P <0.05).Ratio of proximal and distal gastric area at maximum satiety in PDS group was 1 .63±0.42 and that in healthy control group was 2.33±0.63,and the difference was statistically significant (t =5 .044,P <0.05 ).Ratio of proximal and
distal gastric volume in PDS group and healthy group at fasting was 0.60 ± 0.38 and 0.46 ± 0.19, respectively,that at maximum satiety was in PDS group and healthy group 0.91 ±0.27,1 .30 ±0.48, respectively,and the differences were statistically significant (t = - 1 .505 and 3.970,both P <0.05). The emptying rates of proximal stomach of PDS group at 30 min and 90 min postprandial were (28.10 ± 25 .03)% and (68.80±16.14)%,respectively,those of healthy control group were (34.97 ±14.41 )%and (75 .86 ±9.86)%,and the differences were statistically significant (t =1 .087 and 1 .718,both P <0.05).There was no statistically significant difference in emptying rate of distal stomach between two groups (all P > 0.05 ).Conclusions The volume of distal stomach was larger than that of proximal stomach in PDS patients at maximum satiety in liquid food distribution,which may be the cause of decreasing emptying rate of proximal stomach.And proximal stomach dominated gastric motility in PDS patients.
3.Polydatin activates MEK/ERK signaling pathway to improve ocular surface dysfunction and pathology in rats with scopolamine-induced dry eye disease
Jin CAO ; Lanlan GONG ; Xudong LYU
Chinese Journal of Immunology 2024;40(5):1089-1095
Objective:To explore effect of polydatin on ocular surface dysfunction and pathology in dry eye disease(DED)rats and its protective mechanism.Methods:Eighteen from 90 SD rats were randomly chosen as control group,and other rats were injected subcutaneously with scopolamine hydrobromide to establish DED model.After modeling,rats were separated into model group,0.05%polydatin group,0.5%polydatin group,0.5%polydatin+U0126[mitogen-activated protein kinase kinase(MEK)inhibi-tor]group,with 18 rats in each group;each administration group was given corresponding doses of drugs for intervention,and rats in control group and model group were given same amount of normal saline.On the 7th,14th,21st,and 28th day of intervention,Schirmer test was performed to measure tear secretion amount of rats in each group,fluorescein staining was performed to measure tear film breakup time(BUT)and corneal damage of rats in each group;RT-qPCR was performed to measure mRNA expression of pro-inflammatory factors TNF-α,IFN-γ and IL-1β in cornea and conjunctival tissues;HE staining was performed to observe histopatho-logical changes of cornea;PAS staining was performed to evaluate number of conjunctival goblet cells;Western blot was performed to detect expressions of MEK/extracellular regulatory protein kinase(ERK)pathway related proteins.Results:Compared with control group,tear secretion amount BUT and number of conjunctival goblet cells in model group were obviously reduced(P<0.05),corneal injury score,TNF-α,IFN-γ and IL-1β mRNA levels were obviously increased(P<0.05),corneal epithelial layer was thinned,a large number of inflammatory cell infiltration and new capillaries could be seen;compared with model group,tear secretion amount,BUT,number of conjunctival goblet cells,p-MEK1/2/MEK1/2 and p-ERK1/2/ERK1/2 in 0.05%polydatin group and 0.5%polydatin group were obviously increased(P<0.05),corneal injury score,TNF-α,IFN-γ and IL-1β mRNA levels were obviously reduced(P<0.05),pathological damage of corneal tissue was obviously relieved;U0126 could significantly attenuate protective effects of polydatin on ocular surface dysfunction and pathological changes in DED rats.Conclusion:Polydatin may improve ocular surface dysfunction and pathology of DED rats induced by scopolamine by activating MEK/ERK pathway.
4.Biomechanical study of a self-designed anatomic plate for posterolateral tibial plateau
Xudong CHU ; Bin XU ; Huajun QIAN ; Chunxiao QIAN ; Deping ZHAN ; Jiangshan ZHOU ; Lyu PAN
Chinese Journal of Orthopaedic Trauma 2020;22(11):978-982
Objective:To evaluate the biomechanical performance of our self-designed anatomical plate for posterolateral tibial plateau in comparison with conventional plates for treatment of posterolateral tibial plateau fractures.Methods:A novel anatomic plate for posterolateral tibial plateau was designed according to the data measured in the superior fibular capitulum and 3D CT segmentation. Twenty-four knee joints were obtained from 12 freshly frozen adult cadavers to make models of posterolateral tibial plateau fracture. The models were divided into 3 groups( n=8). In group A, fixation was simulated via the supra-fibular-head approach after autogenous iliac bone-graft by our self-designed anatomic plate for posterolateral tibial plateau; in group B, fixation was simulated via the posterior tibial approach after autogenous iliac bone-graft by a small T-plate; in group C, fixation was simulated via the supra-fibular-head approach after autogenous iliac bone-graft by a normal L-plate. Biomechanical tests were carried out in the 3 groups to measure the vertical displacements of split bone fragment under the vertical compression loads of 500 N, 1,000 N and 1,500 N and the maximum compression upon failure of internal fixation (compressed displacemen t=3 mm). Results:At the vertical compression loads of 500 N, 1,000 N and 1,500 N, the vertical displacements of split bone fragment showed significant differences among the 3 groups ( P<0.05); there was a significant difference between group C and groups A and B, respectively ( P<0.05), but an insignificant difference between group A and group B ( P>0.05) though group A performed slightly better. In terms of the maximum compression upon failure of internal fixation, significant differences existed among the 3 groups ( P<0.05); there was a significant difference between group C and groups A and B, respectively ( P< 0.05), but an insignificant difference between group A and group B ( P>0.05). Conclusions:Our self-designed anatomic plate for posterolateral tibial plateau can firmly fixate the fracture fragments of posterolateral condyle.
5.Pathogenic bacteria and drug sensitivity analysis of chronic suppurative otitis media and cholesteatoma in plateau area
Yun LYU ; Jian HE ; Xudong WEI ; Shaojie ZHANG ; Yurong YAO
Chinese Archives of Otolaryngology-Head and Neck Surgery 2019;26(1):21-24
OBJECTIVE To invest igate the distribution of pathogenic bacteria and the antibiotic susceptibility of otitis media in plateau areaandto guide clinical drug application rationally. METHODS Middle ear secretions were collected from 218 inpatients and outpatients(220 ears) with otitis media in our department from December 2016 to January 2018 and were performed by isolation and identification of pathogenic bacteriaand drug sensitivity test. RESULTS 1. 152 strains of microbes were isolatedincluding 125 casesof bacterial infection and 8 cases of fungal infection. 2. The gram-positive bacteria in middle ear effusions of chronic suppurative otitis media was higher than those of cholesteatoma, of which Staphylococcus aureuswas the most frequently isolated pathogen. While Pseudomonas aeruginosa was the highest in cholesteatoma. 3. The antibiotic sensitivity of pathogenic bacteria varies from strain to strain. CONCLUSION Staphylococcus aureusand Pseudomonas aeruginosa were the main pathogenic bacteria. Common pathogenic bacteria were resistance to penicillin and levofloxacin, which were commonly used in clinic. Therefore, bacterial culture should be carried out and rational drug use should be guided.
6.Meta-analysis of the efficacy of proton pump inhibitors for the laryngopharyngeal reflux disease
Yun LYU ; Jian HE ; Xudong WEI ; Shoukai ZHANG ; Yuan FANG ; Xiaolong YANG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2018;25(3):150-156
OBJECTIVE To explore validity and reliability of proton pump inhibitors (PPI) in adult patients with laryngopharyngeal reflux disease(LPRD). METHODS Randomized and placebo-controlled clinical trials regarding the effectiveness of PPIs on LPRD patients were systematically extracted from PubMed, Cochrane Library, and EMBASE, Web of Science, CNKI, CBM, VIP and WANFANG from the beginning of indexing to August 2017. After selection and quality assessment, data were extracted from eligible studies meeting the exclusion and inclusion criteria. The Review Manager software(version 5.2, The Cochrane Collaboration) was performed for all of the statistical analysis in this meta-analysis. The level of evidence was assessed by the GRADE (Grades of Recommendation, Assessment, Development and Evaluation). RESULTS The sample comprised 14 RCTs including a pooled total of 821 LPRD patients were eligible for this meta-analysis, of which 447 and 374 patients took PPIs and placebo, respectively. Pooled results demonstrated that total RSI significantly improved for patients who received PPI therapy by comparing with those receiving placebo [SDM=2.12, 95%CI(0.80; 3.45), P<0.05]. PPI treatment group showed effective in response rate [OR=1.62, 95%CI (0.89, 2.95), P>0.05], but the differences between PPIs and placebo groups was not statistically significant. CONCLUSION PPI treatment could significantly improve reflux symptoms in LPRD patients, therefore, should be taken into consideration for LPRD management with other strategies, such as lifestyle modification.
7.The effect of taking medication at different times on Mongolian and Han patients with non-dipping hypertension
Jinyu LI ; Guanhua HUANG ; Xudong LYU
China Modern Doctor 2024;62(21):90-94
Objective To investigate the effects of amlodipine benzenesulfonate tablets administered at different time points on blood pressure in Mongolian and Han patients with non-dipping hypertension.Methods A total of 68 cases of Mongolian with non-dipping hypertension who had settled in Inner Mongolia and had not intermarried with other nationalities within three generations and 70 cases of Han patients with non-dipping hypertension were consecutively selected as research object,they all received diagnosis at the Second Affiliated Hospital of Baotou Medical College,Inner Mongolia University of Science and Technology from September 2022 to September 2023,and were divided into two groups on average according to the simple randomization method,the group of medication after dinner(group A)and the group of medication before bedtime(group B),and both groups took irbesartan 75mg in the morning,once a day.After that amlodipine benzenesulfonate tablets 5mg,once a day,were added after dinner in group A and before bedtime in group B.Treatment was carried out for 6 months.The ambulatory blood pressure parameters of total patients,Han patients and Mongolian patients before and after treatment were compared between two groups,as well as the proportion of nonpareil blood pressure and the blood pressure compliance rate of total patients after treatment between two groups.Results ①Dynamic blood pressure indicators:After treatment,the dynamic blood pressure levels in both groups of patients decreased,and the decrease in indicators in group A was more significant than that in group B(P<0.05);Han and Mongolian patients were compared within their respective populations,and the results were consistent with the inter group comparison of the total patients(P<0.05).②Correction of non dipper blood pressure:After treatment,the effective rates of dipper blood pressure in group A were higher than those in group B,and the difference was statistically significant(P<0.05).③Blood pressure compliance rate:The blood pressure compliance rate of group A patients after treatment was higher than that of group B,and the difference was statistically significant(P<0.05).Conclusion Amlodipine benzenesulfonate taken at different times has certain therapeutic effects on patients with non-dipping hypertension,and therapeutic effects are better compared with the effect of patients taking the drug after dinner,the degree of improvement of ambulatory blood pressure indexes,correction rate of non-dipping hypertension and blood pressure compliance rate are better than those of patients who take the drug before bedtime,and the effect of taking the drug after dinner is better than that take the drug before bedtime for both Mongolian and Han patients with non-dipping hypertension.
8.Value of serological marker detection in hierarchical management of Crohn's disease
Jiajia SHEN ; Yihong FAN ; Bin LYU ; Xudong ZHANG
Chinese Journal of Digestion 2018;38(1):14-19
Objective To explore the values of serological markers in the phenotype diagnosis of Crohn's disease (CD).Methods From January 2015 to December 2016,100 patients diagnosed as CD were enrolled.The levels of human anti-Saccharomyces cerevisiae antibody (ASCA) IgG,ASCA IgA,anti-mannobioside carbohydrate antibody (AMCA),anti-chitobioside carbohydrate antibody (ACCA),antibody against outer membrane porin C of Escherichia coli (anti-OmpC),antibody against pseudomonas fluorescens (anti-I2),flagella protein antibody,perinuclear anti-neutrophil cytoplasmic antibodies (pANCA) and antibodies to proteinase-3 (anti-PR3) were detected by indirect enzyme-linked immunosorbent assay (ELISA).Binary Logistic regression analysis was used to analyze the correlation between the serological markers and the phenotype of CD.The diagnosis values were evaluated by receiver operating characteristic (ROC) curve.Results The level of anti-I2 was significantly correlated with the narrow type of patients with CD (odd ratios (OR) =34.304,P=0.011);ACCA and flagella protein antibody were significantly correlated with the penetrating type of patients with CD (OR =1.024,P=0.027;OR=2.702,P=0.021).ASCA IgA and AMCA were significantly correlated with the lesion involved gastric,duodenum and small intestine in CD patients (OR =1.146,P=0.044;OR=1.035,P=0.013).In the diagnosis of narrow type CD,the diagnostic accuracy of anti-I2 was the highest (area under curve (AUC) =0.631,P=0.043),and according to this result,its best cutoff value was 0.650,and then the sensitivity increased to 28.6% (8/28).In the diagnosis of penetrating type CD,the diagnostic accuracy of flagella protein antibody was the highest (AUC=0.725,P<0.01).Anti-OmpC (AUC =0.677,P=0.006) and ACCA (AUC=0.644,P=0.025) also had higher diagnostic accuracy.According to these results,the best cutoff values of flagella protein antibody,anti-OmpC and ACCA were 0.395,0.225 and 59.500,respectively,and then their sensitivity improved after correcting the cutoff values.The AUC of any one item positive in the combination of flagella protein antibody,anti-OmpC and ACCA in the diagnosis of penetrating type CD was 0.761 (P<0.01),which was significantly higher than the diagnostic accuracy of single antibody detection.In the diagnosis of the lesions involved gastric,duodenum and small intestine,the diagnostic accuracy of AMCA was the highest,and its AUC was 0.817 (P<0.01).ASCA IgA (AUC=0.772,P=0.001) and ACCA (AUC=0.752,P=0.001) also had higher diagnostic accuracy.The AUC of any one item positive of ASCA IgA and AMCA in the diagnosis of CD lesions involved gastric,duodenum and small intestine was 0.905 (P<0.01),which was significantly higher than the diagnostic accuracy of single antibody detection.The best cutoff values of ASCA IgA,AMCA and ACCA were 21.500,55.000 and 29.500,respectively.And their sensitivity improved after correcting the cutoff values.The sensitivity of any one item positive in the three items also improved.Conclusion The detection of serological markers has great clinical values in the hierarchical management of phenotypes in Chinese CD population.
9.Comparison of Blood Routine Changes in Treatment of Aplastic Anemia by Kidney-tonifying and Blood-generating Method and Qi-promoting and Blood-nourishing Method and Mode of Treatment by Stages
Yan LYU ; Rui LI ; Xudong TANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(14):73-79
ObjectiveTo explore the clinical effect of kidney-tonifying and blood-generating method and qi-promoting and blood-nourishing method combined with western medicine on the treatment of aplastic anemia and the characteristics of blood routine recovery, and to explore a new phased treatment model for aplastic anemia. MethodThis study was based on a prospective, multicenter, double-blind, and randomized controlled clinical trial. Patients with aplastic anemia from 19 centers were analyzed and divided into a kidney-tonifying and blood-generating group and a Qi-promoting and blood-nourishing group, which were treated with traditional Chinese medicine (TCM) combined with western medicine. The clinical effect and the changes in blood routine in each group during treatment were evaluated. ResultDuring the observation period, 375 cases of aplastic anemia were included in two groups, and TCM syndrome differentiation conformed these cases as Qi-deficiency type and both Qi and blood-deficiency type. These cases were randomly divided into two groups, including 184 in the kidney-tonifying and blood-generating group and 191 in the Qi-promoting and blood-nourishing group, being treated by kidney-tonifying and blood-generating granules and Qi-promoting and blood-nourishing granules, respectively, and combined oral androgen and ciclosporin soft capsules. The treatment lasted for six months and was divided into three stages. Visits were conducted from the beginning of the treatment to the end of the first, fourth, and sixth months. The curative effect was evaluated six months later. The total effective rate of the kidney-tonifying and blood-generating group was 86.4% (159/184), which was significantly better than that of the Qi-promoting and blood-nourishing group [68.6% (131/191), P<0.01)]. The results of the percentage quartile of blood cell growth in each stage of the 2 groups were analyzed. The hemoglobin concentration and platelet count of the patients in the kidney-invigorating blood group continued to increase after treatment, and significantly increased in the second and third stages compared with the first stage (P<0.05). The increase of reticulocyte count was most significant in the first stage of treatment (P<0.05). The reticulocyte count in supplementing Qi and nourishing blood group increased significantly in the first and second stages of treatment (P<0.05). The other observation indicators increased at each stage, but there was no statistical difference in the growth rate. The effects of the two groups were compared by stages. In the second stage of treatment, the increase of hemoglobin concentration in the kidney-invigorating blood group was better than that in the supplementing Qi-nourishing blood group (P<0.05). The increase of platelet count and red blood cell count in supplementing Qi and nourishing blood group was greater (P<0.05). In the third stage of treatment, the increase of hemoglobin concentration in the bushen Shengxue group was more significant (P<0.05). ConclusionThe overall effective rate of the kidney-tonifying and blood-generating method in the treatment of aplastic anemia is better than that of the Qi-promoting and blood-nourishing method, with significant effects and safety. This study has proposed a three-stage early treatment mode for aplastic anemia. The first and third stages (0-1, 5-6 months) were mainly treated by invigorating kidney and generating blood. In the second stage of treatment (2-4 months), invigorating kidney and generating blood combined with invigorating Qi and nourishing blood were adopted. It may be closer to the actual clinical treatment response and objective rule changes of aplastic anemia.
10.Comparison of survival in elderly patients with acute myeloid leukemia treated with oral arsenic-containing Qinghuang Powder and low intensive chemotherapy
Yan LYU ; Weiyi LIU ; Richeng QUAN ; Haiyan XIAO ; Xudong TANG ; Chi LIU ; Liu LI ; Hongzhi WANG ; Yonggang XU ; Xiaoqing GUO ; Teng FAN ; Xiaomei HU
Journal of Leukemia & Lymphoma 2018;27(7):396-399
Objective To investigate the survival of oral arsenic-containing Qinghuang Powder (QHP) and low intensive chemotherapy (LIC) in the treatment of elderly patients with acute myeloid leukemia (AML).Methods Forty-two AML patients older than 60 years in Xiyuan Hospital from January 2015 to December 2017 were retrospectively analyzed.Of them,20 cases were treated with QHP (QHP group),22 cases were treated with LIC (LIC group).The survivals of the two groups were compared.Results There was no significant difference of median survival time (13 months vs.13.5 months,x2 =0.096,P =0.757),1-year survival rates (59.1% vs.70.0 %,x2 =0.543,P =0.461),2-year survival rates (13.6 % vs.15.0 %,x2 =0.016,P > 0.05),and 3-year survival rates (4.6 % vs.5.0 %,x2 =0.005,P > 0.05) between LIC and QHP groups.There was no significant difference of median survival time in age ≥75 year (12 months vs.12.5 months,x2 =1.317,P =0.251),performance status scores > 2 (12 months vs.12 months,x2 =0.834,P =0.361),hematopoietic stem cell transplantation with combined disease index > 2 (12 months vs.13 months,x2 =1.726,P =0.189),secondary AML (10 months vs.14 months,x2 =1.552,P =0.213),and poor cytogenetics (12 months vs.8 months,x2 =0.479,P =0.489) between LIC and QHP group.Conclusion The survival of elderly AML patients is considerable in patients treated with oral QHP and LIC,which suggests that oral QHP may be an equivalent alternative treatment since elderly AML (especially more than 75 years) patients refused to LIC therapy.