1. Comprehensive Evaluation and Application of Experimental Sources of Variation in Gut Microbiome Sequencing Studies
Ke-Lin XU ; Yue ZHUANG ; Si-Bo ZHU ; Jiang-Li XUE ; Yan-Feng JIANG ; Zi-Yu YUAN ; Chen SUO ; Tie-Jun ZHANG ; Ming LV ; Xing-Dong CHEN ; Si-Bo ZHU ; Yan-Feng JIANG ; Jiu-Cun WANG ; Xing-Dong CHEN ; Si-Bo ZHU ; Yan-Feng JIANG ; Xing-Dong CHEN ; Chen SUO ; Tie-Jun ZHANG ; Ming LV
Chinese Journal of Biochemistry and Molecular Biology 2022;38(7):959-970
		                        		
		                        			
		                        			 Gut microbiome sequencing studies have great potential to translate microbial analysis outcomes into human health research. Sequencing strategies of 16S amplicon and whole-metagenome shotgun (WMS) are two main methods in microbiome research with respective advantages. However, how sample heterogeneity, sequencers and library preparation protocols affect the sequencing reproducibility of gut microbiome needs further investigation. This study aims to provide a reference for the selection of sequencing technologies by comparing differences in microbial composition from different sampling sites. The results of three widely adopted sequencers showed that the technical repetition correlation (r= 0. 94) was high in WMS method, while the biological repetition correlation (r = 0. 69) was low. Bray-Curtis distance identified that dissimilarity from biological replicates was larger than that of technical replicates (P<0. 001). In addition, dissimilarity and specific taxonomic profiles were observed between 16S and WMS datasets. Our results imply that homogenization is a necessary step before sample DNA extraction. The sequencers contributed less to taxonomic variation than the library preparation protocols. We developed an empirical Bayes approach that " borrowed information" in calculations and analyzed batch effect parameters using standardized data and prior distributions of (non-) parameters, which may improve population comparability between 16S and WMS and provide a basis for further application to fusion analysis of published 16S and microbial datasets. 
		                        		
		                        		
		                        		
		                        	
2.Studies on wnt5a inducing differentiation of cardiac progenitor cells to myocardial cells
Liang-Yu WU ; Ping XIANG ; Ling-Juan LIU ; Tong-Chuan HE ; Song-Hua FANG ; Tie-Wei LV ; Jie TIAN ; Mi LI
Medical Journal of Chinese People's Liberation Army 2018;43(4):316-321
		                        		
		                        			
		                        			Objective To investigate the effect of Wnt5a on inducing differentiation of Cp15-5a cell to myocardial cell.Methods Recombinant adenovirus wnt5a (Ad-wnt5a) and Ad-GFP was amplified with human embryo kidney 293 cells (HEK293 cells),and then transfected into CP15-5a cells and 3 experiment groups were set up:wnt5a group,GFP group and blank control group.Flow cytometry was used to detect the transfection efficiency of Ad-wnt5a and Ad-GFP.One week after transfection,the expressions of genes GATA binding protein 4 (GATA4) and myocardial enhancement factor 2C (MEF2C) were analyzed by realtime quantitative PCR (qRT-PCR).Two weeks after transfection,the expressions of cardiac-specific connexin 43 (Cx43) and cardiac troponin T (cTnT) in Ad-wnt5a-induced CP15-5a cells were detected by Western blotting and immunofluorescence techniques.The sodium current expression (INa) was detected by whole cell patch clamp techniques.Results The transfection efficiency of Ad-wnt5a and Ad-GFP was 42.8% and 44.3%,respectively.One week after transduction,the expressions of GATA4 and MEF2C were significantly higher in wnt5a group (1.717 ± 0.220 and 1.847 ± 0.190) than in GFP group (1.003 ± 0.087 and 0.456 ± 0.042,P<0.05) and blank control group (0.961 ± 0.063 and 0.500 ± 0.095,P<0.05),while no significant difference existed between GFP group and blank control group.Two weeks after transduction,the expressions of CX43 and cTnT were significantly higher in wnt5a group (1.597 ± 0.267 and 0.727 ± 0.100) than in GFP group (0.723 ± 0.047 and 0.217 ± 0.021,P<0.05) and blank control group (0.783 ± 0.1333 and 0.253 ± 0.102,P<0.01),while no significant difference existed between GFP group and blank control group.INa was detected in the wnt5a group compared with GFP group and blank control group.Conclusion wnt5a may induce differentiation of cp 15-5a cell into myocardial cell.
		                        		
		                        		
		                        		
		                        	
3.Impact of premature rupture of membranes on neonatal complications in preterm infants with gestational age <37 weeks.
Shun-Yan DUAN ; Xiang-Yong KONG ; Feng-Dan XU ; Hong-Yan LV ; Rong JU ; Zhan-Kui LI ; Shu-Juan ZENG ; Hui WU ; Xue-Feng ZHANG ; Wei-Peng LIU ; Fang LIU ; Hong-Bin CHENG ; Yan-Jie DING ; Tie-Qiang CHEN ; Ping XU ; Li-Hong YANG ; Su-Jing WU ; Jin WANG ; Li PENG ; Xiao-Lin ZHAO ; Hui-Xian QIU ; Wei-Xi WEN ; Ying LI ; Lan LI ; Zheng WEN ; Guo GUO ; Feng WANG ; Gai-Mei LI ; Wei LI ; Xiao-Ying ZHAO ; Yun-Bo XU ; Wen-Chao CHEN ; Huan YIN ; Xiao-Liang WANG ; Rui-Yan SHAN ; Mei-Ying HAN ; Chun-Yan YANG ; Zhi-Chun FENG
Journal of Southern Medical University 2016;36(7):887-891
OBJECTIVETo investigate the effect of premature rupture of the membrane (PROM) on neonatal complications in premature infants.
METHODSThe registration information of 7684 preterm infants with gestational age <37 weeks were collected from the cooperative units in the task group between January 1, 2014 to December 31, 2014. Specially trained personnel from each cooperative units filled in the unified form in a standardized format to record the gender, gestational age, birth weight, PROM, placental abruption, antenatal corticosteroid, Apgar score, amniotic fluid pollution, and complications of the infants. The data were analyzed comparatively between the cases with PROM and those without (control).
RESULTSThe preterm mortality rate was significantly lower but the incidences of ICH, NEC, ROP and BPD were significantly higher in PROM group than in the control group (P<0.05). The 95% confidence interval of the OR value was <1 for mortality, and was >1 for ICH, NEC, ROP and BPD. After adjustment for gestational age, birth weight, gender, mode of delivery, placental abruption, placenta previa, prenatal hormones, gestational diabetes mellitus (GDM), gestational period hypertension and 5-min Apgar score <7, the incidences of NEC, ROP and BPD were significantly different between the two groups (P<0.05) with 95% confidence interval of OR value >1, but the mortality rate and incidence of ICH were not significantly different between the two groups (P>0.05).
CONCLUSIONPROM is a risk factor for NEC, ROP and BPD in preterm infants, and adequate intervention of PROM can reduce the incidences of such complications as NEC, ROP and BPD in the infants.
Apgar Score ; Birth Weight ; Female ; Fetal Membranes, Premature Rupture ; pathology ; Gestational Age ; Humans ; Incidence ; Infant, Newborn ; Infant, Newborn, Diseases ; etiology ; Infant, Premature ; Pregnancy ; Risk Factors
4.Protective effect of ginseng co-enzyme Q10 suncream on the ultraviolet radiation-induced skin damage in mice
Haiyou WU ; Chuqun QIU ; Meiting LIANG ; Jingbin ZHENG ; Ziwei HU ; Simin LV ; Tie WU
Acta Laboratorium Animalis Scientia Sinica 2016;24(6):601-606
		                        		
		                        			
		                        			Objective To explore the effect of ginseng co?enzyme Q10 suncream on the skin damage caused by ul?traviolet ( UV) radiation in mice. Methods 36 mice were randomly assigned to four groups. The mice were shaved on the back and the left untreated side was taken as control group, or was treated with UV as model group. Before treated with UV, the mice were painted with suncream containing ginseng co?enzyme Q10 , or octyl methoxycinnamate as positive con?trols. The mice were treated for 8 weeks. At the end of the experiment, blood samples of all mice were collected from the eyes, then subjected to cell counting or biochemical measurements, and skin samples were cut for pathological examina?tion. Results Compared with the control group, there was a significant increase in white blood cell counts ( P<0?05 ) and MDA content ( P<0?05 ) , and declined serum levels of SOD ( P <0?05 ) and GSH?Px ( P <0?05 ) in the model group, and the skin was rough and wrinkled with stratum corneum exfoliation. Compared with the model group, the mice of ginseng co?enzyme Q10 suncream group had significantly lower white blood cell count ( P<0?05 ) and MDA content ( P<0?05), and increased serum levels of T?SOD(P<0?05) and red blood cell counts (P<0?05). The skin had no rough? ness and wrinkles and without stratum corneum exfoliation. Compared with the model group, the positive control group showed significantly decreased white blood cell count (P<0?05) and MDA content (P<0?05), and increased serum lev?els of GSH?Px(P<0?05). The skin had no roughness and wrinkles and no stratum corneum exfoliation. However, there was no significant difference between the ginseng co?enzyme Q10 suncream group and positive control group. Conclusions Ginseng co?enzyme Q10 suncream shows satisfactory preventive effects on the UV radiation?induced skin damage in mice, similar to the preventive effects of the octyl methoxycinnamate?containing sunsream.
		                        		
		                        		
		                        		
		                        	
5.Expression and clinical significance of IL-23 and IL-17 in patients with colorectal cancer
Jiansheng WANG ; Yuanpeng LV ; Juntao MENG ; Jinxiao SONG ; Lizhi ZHANG ; Yanqing TIE
Chinese Journal of Clinical Oncology 2014;(9):580-584
		                        		
		                        			
		                        			Objective:This study was conducted to investigate the levels of IL-23 and IL-17 in sera and tumor tissues and to ana-lyze their association with clinico-pathological parameters in patients with colorectal cancer (CRC). Methods:A total of 24 healthy sub-jects and 56 patients with CRC were recruited in this study. Enzyme-linked immunosorbent assay was performed to measure the concen-trations of IL-23 and IL-17 in the sera of CRC patients and healthy controls and in cultured normal tissues and tumor tissues from CRC patients. Results:The levels of serum IL-23 and IL-17 were significantly higher in CRC patients than in healthy controls (P<0.01). The supernatant levels of IL-23 and IL-17 in tumor tissues were significantly higher than in normal tissues (P<0.05). Serum IL-17 level was correlated with tumor location, lymph node metastasis, and TNM stage (P<0.05), whereas serum IL-23 level showed no correlation with all clinico-pathological parameters (P>0.05). Tumor supernatant IL-17 level was correlated with tumor differentiation, lymph node metastasis, and TNM stage (P<0.05), whereas tumor supernatant IL-23 level was correlated only with TNM stage (P<0.05). Conclu-sion:IL-23 and IL-17 may be involved in the immuno-pathogenesis of CRC progression.
		                        		
		                        		
		                        		
		                        	
6.The observation and evaluation of bone histomorphometry in three osteoporosis models of mice
Yongli SITU ; Xing KE ; Yi LIU ; Simin LV ; Yuechun ZHONG ; Liao CUI ; Tie WU
Chinese Pharmacological Bulletin 2014;(9):1281-1286,1287
		                        		
		                        			
		                        			Aim To investigate whether D-galactose cause osteoporosis and the difference compared with the osteoporosis induced by ovariectomy, and to deter-mine whether ovariectomy coupled with D-galactose ac-celerated the progress of osteoporosis and whether es-trogen had a preventive effect on these osteoporosis models. Methods Sixty SPF mice were randomly divided into six groups , namely sham-operated group, D-galactose group, OVX group, OVX + D-galactose group, OVX + D-galactose + diethylstilbestrol group and D-galactose + diethylstilbestrol group. Seventy days later, the right tibia was processed with undecal-cified sections for bone histomorphometric analysis. Results Compared with the sham-operated group, %Tb. Ar, Tb. Th and Tb. N decrease by 50. 4%, 25. 4%, 50. 9% ( P <0.01 ) respectively, Tb. Sp in-creased by 169. 4% (P <0.05), Oc. pm, Oc. No. ,%Oc. S, Oc. N/mm which reflected bone absorption significantly increased ( P < 0.01 ) , and % L. Pm, MAR, BFR/TV, BFR/BV, BFR/BS which reflected bone formation significantly decreased ( P <0.01 ) in OVX group. %Tb. Ar decreased by 30. 4% in D-ga-lactose group, but there was no statistically significant difference. However, the four parameters reflected the bone absorption in D-galactose group increased signifi-cantly ( P<0.05 ) , while the four parameters reflected bone formation decreased significantly ( P < 0.05 ) . OVX+D-galactose group has obvious performance of osteoporosis, but there was no significant difference compared to OVX group, nor to D-galactose group. Estrogen had significant preventive effect on related pa-rameters of osteoporosis induced by D-galactose and o-variectomy coupled with D-galactose. ConclusionsOsteoporosis model of mice can be established by OVX, D-galactose and OVX +D-galactose. Estrogen can effectively prevent bone loss induced by D-galac-tose and OVX+ D-galactose.
		                        		
		                        		
		                        		
		                        	
7.Comparison on throat swabs and respiratory tract aspirates for the detection of respiratory viruses from patients with pneumonia.
Ai-hua LI ; Weng-feng SHI ; Tie-gang ZHANG ; Min LV ; Meng CHEN ; Fang HUANG ; Jiang WU
Chinese Journal of Epidemiology 2013;34(10):1047-1047
		                        		
		                        		
		                        		
		                        			Bodily Secretions
		                        			;
		                        		
		                        			virology
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		                        			Humans
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		                        			Pharynx
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		                        			virology
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		                        			Pneumonia
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		                        			diagnosis
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		                        			virology
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		                        			Respiratory System
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		                        			virology
		                        			
		                        		
		                        	
8.The effects of inhaled budesonide on interleukin-8, C-reactive protein and procalcitonin in bronchoalveolar lavage fluid
Lijun YING ; Tie LV ; Di LU ; Yaosheng MAO
Chinese Journal of Emergency Medicine 2011;20(9):976-980
		                        		
		                        			
		                        			ObjectiveTo investigate the effects of budesonide inhaled on the levels of interleukin-8 (IL-8), C-reactive protein (CRP) and procalcitonin (PCT) in bronchoalveolar lavage fluid (BALF) of patients with AECOPD treated by using inhalation of budesonide in different doses or by using injection of dexamethasone. MethodsNinety AECOPD patients with mechanical ventilation in ICU ward were enrolled from Jan. 2008 through Sep. 2010. All patients were selected in this study as per the criteria of AECOPD set by the Chinese Medical Association. Ninety AECOPD patients treated with routine therapy were randomly (random number) divided into 3 groups. The patients of group A were given budesonide 2 mg inhaled 1 time/d. The patients of group B was given budesonide 4 mg inhaled 1 time/d. The patients of group C had dexamethasone 2. 5 mg injected 1 time/12h. The changes of IL-8, PCT and CRP in both BALF and serum were respectively detected in three groups at the beginning of treatment, and 3 days and 7days after treatment. The detected data of variables were analyzed by SPSS 13.0 package. ResultsThe level of IL-8 in BALF declined in 3days and 7days after treatment, and the magnitude of decrease in the following order:Group B ( budesonide 4mg/d) > Group A ( budesonide 2 mg/d) > Group C ( intravenous dexamethasone)with significant differences among them ( P < 0. 05 ). The decrease in IL-8 in serum showed the similar trend, but there were no statistical differences among them. The CRP and PCT in both BALF and serum had no significant changes. There was a correlation between the concentration of IL-8 in BALF and the duration of mechanical ventilation.Conclusions The treatment with the inhalation of budesonide could more significantly reduced the IL-8 level in BALF compared with intravenous dexamethasone, whereas the CRP and PCT were not changed. The dynamic changes of IL-8 in BALF might be used as an indicator of prognosis in AECOPD patients with mechanical ventilation.
		                        		
		                        		
		                        		
		                        	
9.Effects of Xuebijing injection on serum protein level in early phase of septic rats.
Xuedong SUN ; Di LU ; Tie LV ; Yaosheng MAO
China Journal of Chinese Materia Medica 2010;35(2):223-225
OBJECTIVETo observe the effect of Xuebijing injection on serum protein level in the early phase of septic rats and explain the mechanism from the perspective of molecular biology.
METHODFifty-four healthy wistar rats were randomly divided into control group, sepsis group and Xuebijing treatment group. The rat model of sepsis was established with injecting lipopolysaccharide (LPS) through caudal vein. Serum total protein (TP) and albumin (ALB) were measured at the point of 6, 12 and 24 h with the established model. The expression of AMPK, eEF2 protein in liver in the three groups were detected by Western blot analysis.
RESULTCompared with control gruop, the concentrations of TP and ALB of sepsis group and Xuebijing group were no significant difference with 6, 12 h treatment TP and ALB in sepsis group was lower (P<0.01) than control group after 24 h, and the expression of phos-AMPK, pho-eEF2 protein in livers was increased (P<0.01) simultaneously. All measured indexes in Xuebijing group has no difference with control group. Compared with sepsis group, TP and ALB of Xuebijing group was significantly higher (P<0.05), but litle lower than control group, the expression of phos-AMPK, pho-eEF2 protein in livers was decreased (P<0.05) simultaneously.
CONCLUSIONThese data suggest that Xuebijing injection prohibits catabolising serum albumin and inhibit liver protein catabolism by method of AMPK way.
Animals ; Blood Proteins ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Injections ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Sepsis ; blood ; drug therapy ; pathology
10.An investigation of eperythrozoon infection in a village Liaoning province.
Ying ZHANG ; Zhi- lun ZHANG ; Jie-ying YIN ; Jie LV ; Xiao-chun DONG ; Tie- feng SHEN ; Dun LI
Chinese Journal of Epidemiology 2010;31(7):831-833
		                        		
		                        		
		                        		
		                        			Adolescent
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		                        			Adult
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		                        			Aged
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		                        			Animals
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		                        			Case-Control Studies
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		                        			Child
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		                        			Child, Preschool
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		                        			China
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		                        			epidemiology
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		                        			Ecosystem
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		                        			Female
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		                        			Humans
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		                        			Male
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		                        			Middle Aged
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		                        			Mycoplasma
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		                        			isolation & purification
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		                        			pathogenicity
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		                        			Mycoplasma Infections
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		                        			epidemiology
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		                        			Rural Population
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		                        			Young Adult
		                        			
		                        		
		                        	
            
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