Objective To understand the membrane protein molecular epidemiology of carbapenem-resistant Pseudomonas aeruginosa (CRPA) in the region,and provide some evidence for rational drug use or application of efflux pump inhibitors. Methods Collected Pseudomonas aeruginosa isolated from four hospitals in the region from October 2022 to August 2023,and used SYBR-PCR method to quantitatively detect the relative mRNA expression (RE) levels of 10 membrane protein coding genes,including mexA,B,C,D,E,F,X,Y,and oprD,M. Then categorized the strains into five groups based on ceftazidime,cefepime,imipenem,and meropenem resistance phenotype combination,including the compassionate group (Group Ⅰ),Group Ⅱ with full resistance,IPM,MEM resistant,CAZ and CFP sensitive groups (Group Ⅲ),IPM resistance,MEM non-resistance (sensitive or intermediate) group (Group Ⅳ),IPM,MEM resistance,CAZ and CFP non-resistance groups (Group V).The median RE of each membrane protein-coding gene was analyzed. Results A total of 108 strains of Pseudomonas aeruginosa were collected,with 24 strains in Group Ⅰ as controls and 84 strains in the carbapenem resistant group,including 32 strains in Group Ⅱ,22 strains in Group Ⅲ,13 strains in Group Ⅳ,and 17 strains in Group Ⅴ. The expression of mexD,mexE,mexF,mexX and mexY in the drug-resistant group was higher than that in the control group,and the differences were statistically significant (U=409.5～661.0,all P<0.05). There was no statistically significant difference in mexA,mexB,mexC,oprD and oprM with the control group (U=767.0～1004.5,all P>0.05). There was no significant difference in the expression of RE genes encoding various membrane proteins among strains from different hospitals (H=0.914～7.407,all P>0.05). Among the four different phenotypes,there was no statistically significant difference in the irregular distribution of mexA and oprM RE between each group and the control group (UmexA=95.0～264.0,UoprM=143.0～331.0). The mexC RE in each group was lower than that in the control group,but the differences were not statistically significant (U=134.0～344.5,all P>0.05). MeixE and meixY RE were both higher than the control group,and the differences were statistically significant (UmexE=48.0～230.0,UmexY=83.0～184.0). MeixB was lower than the control group in group Ⅳ (U=72.0),and the differences were statistically significant (all P<0.05). MeixD and meixF showed consistent expression,with higher expression in groups Ⅲ,Ⅳ and Ⅴ compared to the control group (UmeixD=34.0～102.0,UmeixF=65.0～113.0). MeixX was expressed higher in groups Ⅱ,Ⅳ and Ⅴ compared to the control group (U=164.0,58.0,111.0),while oprD was only expressed lower in group Ⅲ than in the control group (U=140.0),with statistically significant differences (all P<0.05). Although the expression of oprD in groups Ⅱ,Ⅳ and Ⅴ was lower than that in the control group,the differences were not statistically significant (U=381.0,102.0,144.0,all P>0.05). Conclusion ExCD,mexEF and mexXY are the main membrane protein combinations of CRPA efflux pumps in Kunming area. Upregulation of mexD,E,F,X,and Y membrane protein expression enhanced efflux. The correlation between mexAB oprM efflux pump and carbapenem resistance in CRPA in this area was low. The low expression of oprD played a role in the efflux mechanism in strains that do not produce β-lactase,but there was no significant difference in low expression in enzyme producing strains.

Objective To determine the expression of type Ⅲ interferon(IFNL)and IFNL receptor 1(IFNLR1)in the genital tract in rhesus macaques to elucidate the biological characteristics of rhesus macaque IFNL system and promote the use of these monkeys in studies on the pathology,drugs and vaccines for human diseases.Methods IFNL1,IFNL3,and IFNLR1 mRNA levels in normal and simian-human immunodeficiency virus(SHIV)/simian immunodeficiency virus(SIV)-infected genital tract tissues of rhesus macaques were detected by gene-specific reverse transcription-polymerase chain reaction,and IFNLR1 immunoreactivity was examined by confocal microscopy.Results IFNL1 and IFNL3 mRNA levels in the uterus and vagina in normal rhesus macaques were below the measurement threshold,but mRNA levels of IFNLR1 and its variant lacking exon 6 could be readily detected.IFNLR1 immunoreactivity was primarily detected in the covering or glandular epithelia of the uterus and vagina.IFNL1 and IFNL3 mRNA levels were increased in some SHIV/SIV-infected macaques,while mRNA levels of IFNLR1 were unchanged and mRNA levels of IFN-stimulated genes(Mx1,Mx2,and OAS)were significantly increased.Conclusions These data indicate that the type Ⅲ IFN system is expressed in the reproductive tract of rhesus macaques,as in humans,thus providing a scientific basis for the use of rhesus macaques in studies of human infectious diseases,such as AIDS.

Objective To determine the expression of type Ⅲ interferon(IFNL)and IFNL receptor 1(IFNLR1)in the genital tract in rhesus macaques to elucidate the biological characteristics of rhesus macaque IFNL system and promote the use of these monkeys in studies on the pathology,drugs and vaccines for human diseases.Methods IFNL1,IFNL3,and IFNLR1 mRNA levels in normal and simian-human immunodeficiency virus(SHIV)/simian immunodeficiency virus(SIV)-infected genital tract tissues of rhesus macaques were detected by gene-specific reverse transcription-polymerase chain reaction,and IFNLR1 immunoreactivity was examined by confocal microscopy.Results IFNL1 and IFNL3 mRNA levels in the uterus and vagina in normal rhesus macaques were below the measurement threshold,but mRNA levels of IFNLR1 and its variant lacking exon 6 could be readily detected.IFNLR1 immunoreactivity was primarily detected in the covering or glandular epithelia of the uterus and vagina.IFNL1 and IFNL3 mRNA levels were increased in some SHIV/SIV-infected macaques,while mRNA levels of IFNLR1 were unchanged and mRNA levels of IFN-stimulated genes(Mx1,Mx2,and OAS)were significantly increased.Conclusions These data indicate that the type Ⅲ IFN system is expressed in the reproductive tract of rhesus macaques,as in humans,thus providing a scientific basis for the use of rhesus macaques in studies of human infectious diseases,such as AIDS.

Objective To understand the membrane protein molecular epidemiology of carbapenem-resistant Pseudomonas aeruginosa (CRPA) in the region,and provide some evidence for rational drug use or application of efflux pump inhibitors. Methods Collected Pseudomonas aeruginosa isolated from four hospitals in the region from October 2022 to August 2023,and used SYBR-PCR method to quantitatively detect the relative mRNA expression (RE) levels of 10 membrane protein coding genes,including mexA,B,C,D,E,F,X,Y,and oprD,M. Then categorized the strains into five groups based on ceftazidime,cefepime,imipenem,and meropenem resistance phenotype combination,including the compassionate group (Group Ⅰ),Group Ⅱ with full resistance,IPM,MEM resistant,CAZ and CFP sensitive groups (Group Ⅲ),IPM resistance,MEM non-resistance (sensitive or intermediate) group (Group Ⅳ),IPM,MEM resistance,CAZ and CFP non-resistance groups (Group V).The median RE of each membrane protein-coding gene was analyzed. Results A total of 108 strains of Pseudomonas aeruginosa were collected,with 24 strains in Group Ⅰ as controls and 84 strains in the carbapenem resistant group,including 32 strains in Group Ⅱ,22 strains in Group Ⅲ,13 strains in Group Ⅳ,and 17 strains in Group Ⅴ. The expression of mexD,mexE,mexF,mexX and mexY in the drug-resistant group was higher than that in the control group,and the differences were statistically significant (U=409.5～661.0,all P<0.05). There was no statistically significant difference in mexA,mexB,mexC,oprD and oprM with the control group (U=767.0～1004.5,all P>0.05). There was no significant difference in the expression of RE genes encoding various membrane proteins among strains from different hospitals (H=0.914～7.407,all P>0.05). Among the four different phenotypes,there was no statistically significant difference in the irregular distribution of mexA and oprM RE between each group and the control group (UmexA=95.0～264.0,UoprM=143.0～331.0). The mexC RE in each group was lower than that in the control group,but the differences were not statistically significant (U=134.0～344.5,all P>0.05). MeixE and meixY RE were both higher than the control group,and the differences were statistically significant (UmexE=48.0～230.0,UmexY=83.0～184.0). MeixB was lower than the control group in group Ⅳ (U=72.0),and the differences were statistically significant (all P<0.05). MeixD and meixF showed consistent expression,with higher expression in groups Ⅲ,Ⅳ and Ⅴ compared to the control group (UmeixD=34.0～102.0,UmeixF=65.0～113.0). MeixX was expressed higher in groups Ⅱ,Ⅳ and Ⅴ compared to the control group (U=164.0,58.0,111.0),while oprD was only expressed lower in group Ⅲ than in the control group (U=140.0),with statistically significant differences (all P<0.05). Although the expression of oprD in groups Ⅱ,Ⅳ and Ⅴ was lower than that in the control group,the differences were not statistically significant (U=381.0,102.0,144.0,all P>0.05). Conclusion ExCD,mexEF and mexXY are the main membrane protein combinations of CRPA efflux pumps in Kunming area. Upregulation of mexD,E,F,X,and Y membrane protein expression enhanced efflux. The correlation between mexAB oprM efflux pump and carbapenem resistance in CRPA in this area was low. The low expression of oprD played a role in the efflux mechanism in strains that do not produce β-lactase,but there was no significant difference in low expression in enzyme producing strains.

Objective:To examine the early outcome of valve sparing aortic root replacement with reimplantation technique (David procedure) with partial upper sternotomy.Methods:From April 2016 to April 2020, 31 patients underwent valve sparing aortic root replacement under partial upper sternotomy at Vascular Surgery Center, Fuwai Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College. There were 28 males and 3 females, aging (44±13) years (range: 11 to 65 years). Preoperative aortic regurgitation was found greater than moderate in 15 patients, moderate in 6 patients and less than moderate in 10 patients. The diameter of aortic annulus was (26±3) mm (range: 21 to 34 mm), the diameter of aortic sinus was (51±6) mm (range: 41 to 68 mm), the diameter of ascending aorta was (43±8) mm (range: 26 to 62 mm). The preoperative ejection fraction was (65±4) % (range: 59% to 72%) and left ventricular end-diastolic diameter was (55±6) mm (range: 42 to 68 mm). All cases were treated with David Ⅰ procedure, including simple David procedure in 26 patients, David+ascending aorta and partial aortic arch replacement in 3 patients, David+thoracic endovascular aortic repair in 1 patient, David+stent elephant trunk implantation in 1 patient.Results:The operation time, cardiopulmonary bypass time and aortic cross-clamping time were (330±58) minutes (range: 214 to 481 minutes), (138±23) minutes (range: 106 to 192 minutes) and (108±17) minutes (range: 82 to 154 minutes), respectively. There were no death and serious complications (stroke, myocardial infarction, renal insufficiency, severe infection, etc.). The postoperative drainage volume within 24 hours was (314±145) ml (range: 130 to 830 ml). The intubation time was (14±3) hours (range: 8 to 21 hours), and the ICU time was ( M( Q R)) 2.1(1.5) days (range: 1.0 to 5.0 days). Eight patients had no blood transfusion, the proportion of red blood cell use was 9.7% (3/31), plasma use was 22.6% (7/31), and platelet use was 71.0% (22/31). The postoperative left ventricular ejection fraction was (62±4)% (range: 54% to 69%), and left ventricular end-diastolic diameter was (48±4) mm (range: 39 to 56 mm). After operation, aortic regurgitation was significantly improved, with no more than moderate regurgitation, small to moderate regurgitation in 3 patients, minor regurgitation in 3 patients, micro regurgitation in 12 patients and no regurgitation in 13 patients. The follow-up period was 3.5(6.1) months (range: 2.0 to 39.0 months). Echocardiographic follow-up data were obtained in 26 cases, including moderate regurgitation in 1 patient, small to moderate regurgitation in 9 patients, minor regurgitation in 5 patients, micro regurgitation in 6 patients and no regurgitation in 5 patients. There were no major adverse cardiovascular events and aortic events during the follow-up period. No patient was reoperated for aortic regurgitation. Conclusion:Valve sparing aortic root replacement under partial upper sternotomy is safe and feasible, and the early result is satisfactory.

Objective:To examine the early outcome of valve sparing aortic root replacement with reimplantation technique (David procedure) with partial upper sternotomy.Methods:From April 2016 to April 2020, 31 patients underwent valve sparing aortic root replacement under partial upper sternotomy at Vascular Surgery Center, Fuwai Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College. There were 28 males and 3 females, aging (44±13) years (range: 11 to 65 years). Preoperative aortic regurgitation was found greater than moderate in 15 patients, moderate in 6 patients and less than moderate in 10 patients. The diameter of aortic annulus was (26±3) mm (range: 21 to 34 mm), the diameter of aortic sinus was (51±6) mm (range: 41 to 68 mm), the diameter of ascending aorta was (43±8) mm (range: 26 to 62 mm). The preoperative ejection fraction was (65±4) % (range: 59% to 72%) and left ventricular end-diastolic diameter was (55±6) mm (range: 42 to 68 mm). All cases were treated with David Ⅰ procedure, including simple David procedure in 26 patients, David+ascending aorta and partial aortic arch replacement in 3 patients, David+thoracic endovascular aortic repair in 1 patient, David+stent elephant trunk implantation in 1 patient.Results:The operation time, cardiopulmonary bypass time and aortic cross-clamping time were (330±58) minutes (range: 214 to 481 minutes), (138±23) minutes (range: 106 to 192 minutes) and (108±17) minutes (range: 82 to 154 minutes), respectively. There were no death and serious complications (stroke, myocardial infarction, renal insufficiency, severe infection, etc.). The postoperative drainage volume within 24 hours was (314±145) ml (range: 130 to 830 ml). The intubation time was (14±3) hours (range: 8 to 21 hours), and the ICU time was ( M( Q R)) 2.1(1.5) days (range: 1.0 to 5.0 days). Eight patients had no blood transfusion, the proportion of red blood cell use was 9.7% (3/31), plasma use was 22.6% (7/31), and platelet use was 71.0% (22/31). The postoperative left ventricular ejection fraction was (62±4)% (range: 54% to 69%), and left ventricular end-diastolic diameter was (48±4) mm (range: 39 to 56 mm). After operation, aortic regurgitation was significantly improved, with no more than moderate regurgitation, small to moderate regurgitation in 3 patients, minor regurgitation in 3 patients, micro regurgitation in 12 patients and no regurgitation in 13 patients. The follow-up period was 3.5(6.1) months (range: 2.0 to 39.0 months). Echocardiographic follow-up data were obtained in 26 cases, including moderate regurgitation in 1 patient, small to moderate regurgitation in 9 patients, minor regurgitation in 5 patients, micro regurgitation in 6 patients and no regurgitation in 5 patients. There were no major adverse cardiovascular events and aortic events during the follow-up period. No patient was reoperated for aortic regurgitation. Conclusion:Valve sparing aortic root replacement under partial upper sternotomy is safe and feasible, and the early result is satisfactory.

OBJECTIVE:To study the effect and its possible mechanism of Xuezhiping capsule on blood lipid levels of high blood lipid model golden hamsters. METHODS:Golden hamsters were randomly divided into normal control group,model group, atorvastatin group (3 mg/kg),combination group (Xuezhiping capsule 1.3 g/kg+atorvastatin 1.5 mg/kg),Xuezhiping capsule low-dose,medium-dose,high-dose groups(Xuezhiping capsule 1.3,2.6,5.2 g/kg),10 in each group. Hamsters in normal control group received normal diet,the other groups were given high-fat diet to establish high blood lipid model. Then relevant drugs were intragastrically given 2 weeks later. Normal control group and model group were intragastrically given equal volume of distilled wa-ter,once a day,for 4 weeks. After last administration,blood lipid indexes(TG,TC,HDL-C,LDL-C,FFA),mRNA and protein expressions of lipid metabolism related genes (PPAR-α,CYP7A1,ACOX,SREBP-1c,ACC1) were detected. RESULTS:Com-pared with normal control group,serum contents of TC,TG,LDL-C,FFA in model group increased,HDL-C content decreased;PPAR-α,CYP7A1,ACOX mRNA and protein expressions were weakened,SREBP-1c,ACC1 mRNA and protein expressions were enhanced(P<0.05). Compared with model group,above-mentioned indexes were obviously improved in Xuezhiping capsule high-dose group,atorvastatin group and combination group (except for HDL-C in Xuezhiping capsule high-dose group)(P<0.05);TC,TG,FFA contents,and PPAR-α,CYP7A1,ACOX,ACC1 mRNA,and CYP7A1,SREBP-1c,ACC1 protein were obviously improved in Xuezhiping capsule medium-dose group(P<0.05);TC,TG contents,and PPAR-α mRNA,and CYP7A1, SREBP-1c protein were obviously improved in Xuezhiping capsule low-dose group (P<0.05). CONCLUSIONS:Xuezhiping cap-sule has a promising effect of lowering blood lipid,the mechanism may be related to activating PPAR-α and inhibiting SREBP-lc signaling pathway.

OBJECTIVETo study the expression of Toll-like receptors (TLRs) mRNA in human trophoblast HTR-8/SVneo cells and the changes in indoleamine 2,3-dioxygenase (IDO) mRNA expression in response to TLR ligand stimulation.

METHODSThe expressions of TLRs and IDO mRNA in human HTR-8/SVneo cells were tested by RT-PCR, and the changes in IDO mRNA levels after exposure to TLR3, TLR4, TLR7/8, and TLR9 ligands were quantitatively analyzed with real-time PCR.

RESULTSIDO and TLR1-10 mRNAs were expressed in HTR-8/SVneo cells. As the cell culture time extended, IDO mRNA expression level tended to increase within 48 h. After stimulation with the TLR ligands, the expression of TLR-3 mRNA was down-regulated while the expression of TLR-4, 7, 8, and 9 mRNA up-regulated. Stimulation of the cells with poly(I:C) lowered the expression of IDO mRNA while IFN-γ increased its expression.

CONCLUSIONSThe expression of IDO mRNA is associated with the nutrition of the maternal-fetal interface. Stimulation with the TLR ligands affects the expression of IDO and TLR mRNA expressions in the cells, which verifies the functional activity of TLRs and suggests a role of IDO in TLR pathway-dependent antiviral immunity.

Cell Line ; Female ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; genetics ; metabolism ; Interferon-gamma ; pharmacology ; Ligands ; Poly I-C ; pharmacology ; RNA, Messenger ; metabolism ; Toll-Like Receptors ; genetics ; metabolism ; Trophoblasts ; cytology ; metabolism

Objective To study the expression of Toll-like receptors (TLRs) mRNA in human trophoblast HTR-8/SVneo cells and the changes in indoleamine 2,3-dioxygenase (IDO) mRNA expression in response to TLR ligand stimulation. Methods The expressions of TLRs and IDO mRNA in human HTR-8/SVneo cells were tested by RT-PCR, and the changes in IDO mRNA levels after exposure to TLR3, TLR4, TLR7/8, and TLR9 ligands were quantitatively analyzed with real-time PCR. Results IDO and TLR1-10 mRNAs were expressed in HTR-8/SVneo cells. As the cell culture time extended, IDO mRNA expression level tended to increase within 48 h. After stimulation with the TLR ligands, the expression of TLR-3 mRNA was down-regulated while the expression of TLR-4, 7, 8, and 9 mRNA up-regulated. Stimulation of the cells with poly(I:C) lowered the expression of IDO mRNA while IFN-γincreased its expression. Conclusions The expression of IDO mRNA is associated with the nutrition of the maternal-fetal interface. Stimulation with the TLR ligands affects the expression of IDO and TLR mRNA expressions in the cells, which verifies the functional activity of TLRs and suggests a role of IDO in TLR pathway-dependent antiviral immunity.

Objective To study the expression of Toll-like receptors (TLRs) mRNA in human trophoblast HTR-8/SVneo cells and the changes in indoleamine 2,3-dioxygenase (IDO) mRNA expression in response to TLR ligand stimulation. Methods The expressions of TLRs and IDO mRNA in human HTR-8/SVneo cells were tested by RT-PCR, and the changes in IDO mRNA levels after exposure to TLR3, TLR4, TLR7/8, and TLR9 ligands were quantitatively analyzed with real-time PCR. Results IDO and TLR1-10 mRNAs were expressed in HTR-8/SVneo cells. As the cell culture time extended, IDO mRNA expression level tended to increase within 48 h. After stimulation with the TLR ligands, the expression of TLR-3 mRNA was down-regulated while the expression of TLR-4, 7, 8, and 9 mRNA up-regulated. Stimulation of the cells with poly(I:C) lowered the expression of IDO mRNA while IFN-γincreased its expression. Conclusions The expression of IDO mRNA is associated with the nutrition of the maternal-fetal interface. Stimulation with the TLR ligands affects the expression of IDO and TLR mRNA expressions in the cells, which verifies the functional activity of TLRs and suggests a role of IDO in TLR pathway-dependent antiviral immunity.