1.Exogenous administration of zinc chloride improves lung ischemia/reperfusion injury in rats.
Shu-Yuan WANG ; Jun-Peng XU ; Yuan CHENG ; Man HUANG ; Si-An CHEN ; Zhuo-Lun LI ; Qi-Hao ZHANG ; Yong-Yue DAI ; Li-Yi YOU ; Wan-Tie WANG
Acta Physiologica Sinica 2025;77(5):811-819
The aim of this study was to investigate the contribution of lung zinc ions to pathogenesis of lung ischemia/reperfusion (I/R) injury in rats. Male Sprague Dawley (SD) rats were randomly divided into control group, lung I/R group (I/R group), lung I/R + low-dose zinc chloride group (LZnCl2+I/R group), lung I/R + high-dose ZnCl2 group (HZnCl2+I/R group), lung I/R + medium-dose ZnCl2 group (MZnCl2+I/R group) and TPEN+MZnCl2+I/R group (n = 8 in each group). Inductively coupled plasma mass spectrometry (ICP-MS) was used to measure the concentration of zinc ions in lung tissue. The degree of lung tissue injury was analyzed by observing HE staining, alveolar damage index, lung wet/dry weight ratio and lung tissue gross changes. TUNEL staining was used to detect cellular apoptosis in lung tissue. Western blot and RT-qPCR were used to determine the protein expression levels of caspase-3 and ZIP8, as well as the mRNA expression levels of zinc transporters (ZIP, ZNT) in lung tissue. The mitochondrial membrane potential (MMP) of lung tissue was detected by JC-1 MMP detection kit. The results showed that, compared with the control group, the lung tissue damage, lung wet/dry weight ratio and alveolar damage index were significantly increased in the I/R group. And in the lung tissue, the concentration of Zn2+ was markedly decreased, while the cleaved caspase-3/caspase-3 ratio and apoptotic levels were significantly increased. The expression levels of ZIP8 mRNA and protein were down-regulated significantly, while the mRNA expression of other zinc transporters remained unchanged. There was also a significant decrease in MMP. Compared with the I/R group, both MZnCl2+I/R group and HZnCl2+I/R group exhibited significantly reduced lung tissue injury, lung wet/dry weight ratio and alveolar damage index, increased Zn2+ concentration, decreased ratio of cleaved caspase-3/caspase-3 and apoptosis, and up-regulated expression levels of ZIP8 mRNA and protein. In addition, the MMP was significantly increased in the lung tissue. Zn2+ chelating agent TPEN reversed the above-mentioned protective effects of medium-dose ZnCl2 on the lung tissue in the I/R group. The aforementioned results suggest that exogenous administration of ZnCl2 can improve lung I/R injury in rats.
Animals
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Reperfusion Injury/pathology*
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Male
;
Rats, Sprague-Dawley
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Rats
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Chlorides/administration & dosage*
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Lung/pathology*
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Zinc Compounds/administration & dosage*
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Apoptosis/drug effects*
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Caspase 3/metabolism*
;
Cation Transport Proteins/metabolism*
2.National bloodstream infection bacterial resistance surveillance report (2022) : Gram-negative bacteria
Zhiying LIU ; Yunbo CHEN ; Jinru JI ; Chaoqun YING ; Qing YANG ; Haishen KONG ; Haifeng MAO ; Hui DING ; Pengpeng TIAN ; Jiangqin SONG ; Yongyun LIU ; Jiliang WANG ; Yan JIN ; Yuanyuan DAI ; Yizheng ZHOU ; Yan GENG ; Fenghong CHEN ; Lu WANG ; Yanyan LI ; Dan LIU ; Peng ZHANG ; Junmin CAO ; Xiaoyan LI ; Dijing SONG ; Xinhua QIANG ; Yanhong LI ; Qiuying ZHANG ; Guolin LIAO ; Ying HUANG ; Baohua ZHANG ; Liang GUO ; Aiyun LI ; Haiquan KANG ; Donghong HUANG ; Sijin MAN ; Zhuo LI ; Youdong YIN ; Kunpeng LIANG ; Haixin DONG ; Donghua LIU ; Hongyun XU ; Yinqiao DONG ; Rong XU ; Lin ZHENG ; Shuyan HU ; Jian LI ; Qiang LIU ; Liang LUAN ; Jilu SHEN ; Lixia ZHANG ; Bo QUAN ; Xiaoping YAN ; Xiaoyan QI ; Dengyan QIAO ; Weiping LIU ; Xiusan XIA ; Ling MENG ; Jinhua LIANG ; Ping SHEN ; Yonghong XIAO
Chinese Journal of Clinical Infectious Diseases 2024;17(1):42-57
Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of national bloodstream infection Bacterial Resistant Investigation Collaborative System(BRICS)were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute(CLSI). WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:During the study period,9 035 strains of Gram-negative bacteria were collected from 51 hospitals,of which 7 895(87.4%)were Enterobacteriaceae and 1 140(12.6%)were non-fermenting bacteria. The top 5 bacterial species were Escherichia coli( n=4 510,49.9%), Klebsiella pneumoniae( n=2 340,25.9%), Pseudomonas aeruginosa( n=534,5.9%), Acinetobacter baumannii complex( n=405,4.5%)and Enterobacter cloacae( n=327,3.6%). The ESBLs-producing rates in Escherichia coli, Klebsiella pneumoniae and Proteus spp. were 47.1%(2 095/4 452),21.0%(427/2 033)and 41.1%(58/141),respectively. The prevalence of carbapenem-resistant Escherichia coli(CREC)and carbapenem-resistant Klebsiella pneumoniae(CRKP)were 1.3%(58/4 510)and 13.1%(307/2 340);62.1%(36/58)and 9.8%(30/307)of CREC and CRKP were resistant to ceftazidime/avibactam combination,respectively. The prevalence of carbapenem-resistant Acinetobacter baumannii(CRAB)complex was 59.5%(241/405),while less than 5% of Acinetobacter baumannii complex was resistant to tigecycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa(CRPA)was 18.4%(98/534). There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of main Gram-negative bacteria resistance among different regions,with statistically significant differences in the prevalence of CRKP and CRPA( χ2=20.489 and 20.252, P<0.001). The prevalence of CREC,CRKP,CRPA,CRAB,ESBLs-producing Escherichia coli and Klebsiella pneumoniae were higher in provinicial hospitals than those in municipal hospitals( χ2=11.953,81.183,10.404,5.915,12.415 and 6.459, P<0.01 or <0.05),while the prevalence of CRPA was higher in economically developed regions(per capita GDP ≥ 92 059 Yuan)than that in economically less-developed regions(per capita GDP <92 059 Yuan)( χ2=6.240, P=0.012). Conclusions:The proportion of Gram-negative bacteria in bloodstream infections shows an increasing trend,and Escherichia coli is ranked in the top,while the trend of CRKP decreases continuously with time. Decreasing trends are noted in ESBLs-producing Escherichia coli and Klebsiella pneumoniae. Low prevalence of carbapenem resistance in Escherichia coli and high prevalence in CRAB complex have been observed. The composition ratio and antibacterial spectrum of bloodstream infections in different regions of China are slightly different,and the proportion of main drug resistant bacteria in provincial hospitals is higher than those in municipal hospitals.
3.National bloodstream infection bacterial resistance surveillance report(2022): Gram-positive bacteria
Chaoqun YING ; Yunbo CHEN ; Jinru JI ; Zhiying LIU ; Qing YANG ; Haishen KONG ; Haifeng MAO ; Hui DING ; Pengpeng TIAN ; Jiangqin SONG ; Yongyun LIU ; Jiliang WANG ; Yan JIN ; Yuanyuan DAI ; Yizheng ZHOU ; Yan GENG ; Fenghong CHEN ; Lu WANG ; Yanyan LI ; Dan LIU ; Peng ZHANG ; Junmin CAO ; Xiaoyan LI ; Dijing SONG ; Xinhua QIANG ; Yanhong LI ; Qiuying ZHANG ; Guolin LIAO ; Ying HUANG ; Baohua ZHANG ; Liang GUO ; Aiyun LI ; Haiquan KANG ; Donghong HUANG ; Sijin MAN ; Zhuo LI ; Youdong YIN ; Kunpeng LIANG ; Haixin DONG ; Donghua LIU ; Hongyun XU ; Yinqiao DONG ; Rong XU ; Lin ZHENG ; Shuyan HU ; Jian LI ; Qiang LIU ; Liang LUAN ; Jilu SHEN ; Lixia ZHANG ; Bo QUAN ; Xiaoping YAN ; Xiaoyan QI ; Dengyan QIAO ; Weiping LIU ; Xiusan XIA ; Ling MENG ; Jinhua LIANG ; Ping SHEN ; Yonghong XIAO
Chinese Journal of Clinical Infectious Diseases 2024;17(2):99-112
Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-positive bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-positive bacteria from blood cultures in member hospitals of National Bloodstream Infection Bacterial Resistant Investigation Collaborative System(BRICS)were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute(CLSI). WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:A total of 3 163 strains of Gram-positive pathogens were collected from 51 member units,and the top five bacteria were Staphylococcus aureus( n=1 147,36.3%),coagulase-negative Staphylococci( n=928,29.3%), Enterococcus faecalis( n=369,11.7%), Enterococcus faecium( n=296,9.4%)and alpha-hemolyticus Streptococci( n=192,6.1%). The detection rates of methicillin-resistant Staphylococcus aureus(MRSA)and methicillin-resistant coagulase-negative Staphylococci(MRCNS)were 26.4%(303/1 147)and 66.7%(619/928),respectively. No glycopeptide and daptomycin-resistant Staphylococci were detected. The sensitivity rates of Staphylococcus aureus to cefpirome,rifampin,compound sulfamethoxazole,linezolid,minocycline and tigecycline were all >95.0%. Enterococcus faecium was more prevalent than Enterococcus faecalis. The resistance rates of Enterococcus faecium to vancomycin and teicoplanin were both 0.5%(2/369),and no vancomycin-resistant Enterococcus faecium was detected. The detection rate of MRSA in southern China was significantly lower than that in other regions( χ2=14.578, P=0.002),while the detection rate of MRCNS in northern China was significantly higher than that in other regions( χ2=15.195, P=0.002). The detection rates of MRSA and MRCNS in provincial hospitals were higher than those in municipal hospitals( χ2=13.519 and 12.136, P<0.001). The detection rates of MRSA and MRCNS in economically more advanced regions(per capita GDP≥92 059 Yuan in 2022)were higher than those in economically less advanced regions(per capita GDP<92 059 Yuan)( χ2=9.969 and 7.606, P=0.002和0.006). Conclusions:Among the Gram-positive pathogens causing bloodstream infections in China, Staphylococci is the most common while the MRSA incidence decreases continuously with time;the detection rate of Enterococcus faecium exceeds that of Enterococcus faecalis. The overall prevalence of vancomycin-resistant Enterococci is still at a low level. The composition ratio of Gram-positive pathogens and resistant profiles varies slightly across regions of China,with the prevalence of MRSA and MRCNS being more pronounced in provincial hospitals and areas with a per capita GDP≥92 059 yuan.
4.Preliminary practice in the postoperative cardiopulmonary exercise testing for children with congenital heart disease.
Xiao Man CAI ; Zhuo Man XU ; Yi Chao XU ; Lin CHEN ; Hao ZHANG
Chinese Journal of Cardiology 2022;50(5):471-479
Objective: To investigate the postoperative cardiopulmonary fitness of children with congenital heart diseases (CHD). Methods: This is a retrospective study. A total of 136 children after cardiac correction undergoing cardiopulmonary exercise test (CPET) in Shanghai Children's Medical Center from March 1 to June 30, 2021 were selected. According to the surgical procedure, the children were divided into two groups: the biventricular correction group (BV group) (n=75) and single ventricular correction group (SV group) (n=61). The BV group was divided into two subgroups: simple congenital heart disease (S-CHD) group (n=35) and complex congenital heart disease (C-CHD) group (n=40). CPET parameters, including VO2 max, VO2/kg max, VO2/kg@AT, O2/HR max, HRR, PetCO2 max, CI, HRR at 1 min, VE/VCO2 slope, OUES/kg and EOV, were analyzed. CPET parameters of patients underwent different procedure were compared. In the subgroup analysis, CPET parameters of patients in S-CHD group and C-CHD group were compared. The parameters of the BV group were compared with the normal value. Linear correlation analysis was used to identify the correlation between the CPET parameters. Results: A total of 136 children with CHD after surgery were enrolled. The age was (9.4±3.1) (ranged 6.2-16.0) years, and there was 84 (61.8%) male. All the children completed CPET examination safely without experiencing serious circulation abnormalities. Compared with the SV group, the VO2/kg max ((32.6±6.9) ml·kg-1·min-1 vs. (23.5±5.9) ml·kg-1·min-1, P<0.001), O2/HR max ((7.24±2.93) ml/beat vs. (6.35±2.17)ml/beat, P=0.030), HRR at 1 min ((32.5±13.9) beat/min vs. (26.3±12.5) beat/min, P=0.036), OUES/kg (36.9±8.8 vs. 29.7±11.8, P=0.001) were significantly higher, VE/VCO2 slope (29.1±5.20 vs. 35.1±8.0, P<0.001) and incidence of EOV (32.0%(24/75) vs. 57.4%(31/61), P=0.027) were significantly lower in BV group. Compared with the simple CHD subgroup, VE/VCO2 slope and the incidence of EOV were higher, VO2/kg max, O2/HR max, HRR at 1 min and OUES/kg were lower in the complex CHD subgroup (all P<0.05). Cardiopulmonary function parameters of the BV group were lower than the normal value. Linear correlation analysis showed that VO2/kg@AT was strongly correlated with VO2/kg max (r=0.86, P<0.001), VO2/kg max was strongly correlated with OUES/kg (r=0.63, P<0.001), HRR was strongly correlated with CI (r=0.91, P<0.001), and VO2/kg max was strongly correlated with OUES/kg (r=0.63, P<0.001). VE/VCO2 slope was strongly correlated with PetCO2 max (r=1.00, P<0.001). Conclusions: The exercise cardiopulmonary function of children after single ventricular correction is weaker than that of biventricular correction, and the exercise tolerance of children after biventricular correction is lower than that of normal children. Among the children after biventricular correction, the exercise tolerance of children with complex CHD is lower than that of children with simple CHD. Postoperative CPET is of important realistic significance for CHD children.
Adolescent
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Child
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China
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Exercise Test/methods*
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Female
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Heart Defects, Congenital/surgery*
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Humans
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Male
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Oxygen Consumption
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Retrospective Studies
5.BRICS report of 2018-2019: the distribution and antimicrobial resistance profile of clinical isolates from blood culture in China
Yunbo CHEN ; Jinru JI ; Chaoqun YING ; Peipei WANG ; Zhiying LIU ; Qing YANG ; Haishen KONG ; Hui DING ; Yongyun LIU ; Haifeng MAO ; Ying HUANG ; Zhenghai YANG ; Yuanyuan DAI ; Guolin LIAO ; Lisha ZHU ; Liping ZHANG ; Yanhong LI ; Hongyun XU ; Junmin CAO ; Baohua ZHANG ; Liang GUO ; Haixin DONG ; Shuyan HU ; Sijin MAN ; Lu WANG ; Zhixiang LIAO ; Rong XU ; Dan LIU ; Yan JIN ; Yizheng ZHOU ; Yiqun LIAO ; Fenghong CHEN ; Beiqing GU ; Jiliang WANG ; Jinhua LIANG ; Lin ZHENG ; Aiyun LI ; Jilu SHEN ; Yinqiao DONG ; Lixia ZHANG ; Hongxia HU ; Bo QUAN ; Wencheng ZHU ; Kunpeng LIANG ; Qiang LIU ; Shifu WANG ; Xiaoping YAN ; Jiangbang KANG ; Xiusan XIA ; Lan MA ; Li SUN ; Liang LUAN ; Jianzhong WANG ; Zhuo LI ; Dengyan QIAO ; Lin ZHANG ; Lanjuan LI ; Yonghong XIAO
Chinese Journal of Clinical Infectious Diseases 2021;14(1):32-45
Objective:To investigate the distribution and antimicrobial resistance profile of clinical bacteria isolated from blood culture in China.Methods:The clinical bacterial strains isolated from blood culture from member hospitals of Blood Bacterial Resistant Investigation Collaborative System (BRICS) were collected during January 2018 to December 2019. Antibiotic susceptibility tests were conducted with agar dilution or broth dilution methods recommended by US Clinical and Laboratory Standards Institute (CLSI). WHONET 5.6 was used to analyze data.Results:During the study period, 14 778 bacterial strains were collected from 50 hospitals, of which 4 117 (27.9%) were Gram-positive bacteria and 10 661(72.1%) were Gram-negative bacteria. The top 10 bacterial species were Escherichia coli (37.2%), Klebsiella pneumoniae (17.0%), Staphylococcus aureus (9.7%), coagulase-negative Staphylococci (8.7%), Pseudomonas aeruginosa (3.7%), Enterococcus faecium (3.4%), Acinetobacter baumannii(3.4%), Enterobacter cloacae (2.9%), Streptococci(2.8%) and Enterococcus faecalis (2.3%). The the prevalence of methicillin-resistant S. aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococcus were 27.4% (394/1 438) and 70.4% (905/1 285), respectively. No glycopeptide-resistant Staphylococcus was detected. More than 95% of S. aureus were sensitive to amikacin, rifampicin and SMZco. The resistance rate of E. faecium to vancomycin was 0.4% (2/504), and no vancomycin-resistant E. faecalis was detected. The ESBLs-producing rates in no carbapenem-resistance E. coli, carbapenem sensitive K. pneumoniae and Proteus were 50.4% (2 731/5 415), 24.6% (493/2001) and 35.2% (31/88), respectively. The prevalence of carbapenem-resistance in E. coli and K. pneumoniae were 1.5% (85/5 500), 20.6% (518/2 519), respectively. 8.3% (27/325) of carbapenem-resistance K. pneumoniae was resistant to ceftazidime/avibactam combination. The resistance rates of A. baumannii to polymyxin and tigecycline were 2.8% (14/501) and 3.4% (17/501) respectively, and that of P. aeruginosa to carbapenem were 18.9% (103/546). Conclusions:The surveillance results from 2018 to 2019 showed that the main pathogens of bloodstream infection in China were gram-negative bacteria, while E. coli was the most common pathogen, and ESBLs-producing strains were in majority; the MRSA incidence is getting lower in China; carbapenem-resistant E. coli keeps at a low level, while carbapenem-resistant K. pneumoniae is on the rise obviously.
6.BRICS report of 2020: The bacterial composition and antimicrobial resistance profile of clinical isolates from bloodstream infections in China
Yunbo CHEN ; Jinru JI ; Chaoqun YING ; Zhiying LIU ; Qing YANG ; Haishen KONG ; Yuanyuan DAI ; Jiliang WANG ; Haifeng MAO ; Hui DING ; Yongyun LIU ; Yizheng ZHOU ; Hong LU ; Youdong YIN ; Yan JIN ; Hongyun XU ; Lixia ZHANG ; Lu WANG ; Haixin DONG ; Zhenghai YANG ; Fenghong CHEN ; Donghong HUANG ; Guolin LIAO ; Pengpeng TIAN ; Dan LIU ; Yan GENG ; Sijin MAN ; Baohua ZHANG ; Ying HUANG ; Liang GUO ; Junmin CAO ; Beiqing GU ; Yanhong LI ; Hongxia HU ; Liang LUAN ; Shuyan HU ; Lin ZHENG ; Aiyun LI ; Rong XU ; Kunpeng LIANG ; Zhuo LI ; Donghua LIU ; Bo QUAN ; Qiang LIU ; Jilu SHEN ; Yiqun LIAO ; Hai CHEN ; Qingqing BAI ; Xiusan XIA ; Shifu WANG ; Jinhua LIANG ; Liping ZHANG ; Yinqiao DONG ; Xiaoyan QI ; Jianzhong WANG ; Xuefei HU ; Xiaoping YAN ; Dengyan QIAO ; Ling MENG ; Yonghong XIAO
Chinese Journal of Clinical Infectious Diseases 2021;14(6):413-426
Objective:To investigate the bacterial composition and antimicrobial resistance profile of clinical isolates from bloodstream infections in China.Methods:The clinical bacterial strains isolated from blood culture were collected during January 2020 to December 2020 in member hospitals of Blood Bacterial Resistant Investigation Collaborative System (BRICS). Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical Laboratory Standards Institute(CLSI, USA). WHONET 5.6 was used to analyze data.Results:During the study period, 10 043 bacterial strains were collected from 54 hospitals, of which 2 664 (26.5%) were Gram-positive bacteria and 7 379 (73.5%) were Gram-negative bacteria. The top 10 bacterial species were Escherichia coli (38.6%), Klebsiella pneumoniae (18.4%), Staphylococcus aureus (9.9%), coagulase-negative Staphylococci (7.5%), Pseudomonas aeruginosa (3.9%), Enterococcus faecium (3.3%), Enterobacter cloacae (2.8%), Enterococcus faecalis (2.6%), Acinetobacter baumannii (2.4%) and Klebsiella spp (1.8%). The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococcus aureus were 27.6% and 74.4%, respectively. No glycopeptide- and daptomycin-resistant Staphylococci were detected. More than 95% of Staphylococcus aureus were sensitive to rifampicin and SMZco. No vancomycin-resistant Enterococci strains were detected. Extended spectrum β-lactamase (ESBL) producing Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis were 48.4%, 23.6% and 36.1%, respectively. The prevalence rates of carbapenem-resistance in Escherichia coli and Klebsiella pneumoniae were 2.3% and 16.1%, respectively; 9.6% of carbapenem-resistant Klebsiella pneumoniae strains were resistant to ceftazidime/avibactam combination. The prevalence rate of carbapenem-resistance in Acinetobacter baumannii was 60.0%, while polymyxin and tigecycline showed good activity against Acinetobacter baumannii. The prevalence rate of carbapenem-resistance of Pseudomonas aeruginosa was 23.2%. Conclusions:The surveillance results in 2020 showed that the main pathogens of bloodstream infection in China were gram-negative bacteria, while Escherichia coli was the most common pathogen, and ESBL-producing strains declined while carbapenem-resistant Klebsiella pneumoniae kept on high level. The proportion and the prevalence of carbapenem-resistant Pseudomonas aeruginosa were on the rise slowly. On the other side, the MRSA incidence got lower in China, while the overall prevalence of vancomycin-resistant Enterococci was low.
7.Gradient pH co-precipitation to recover proteins from plasma discarded in artificial-liver treatment
Bing XU ; Chen PAN ; Xiao-Zhi JIANG ; Man-Zhi LIN ; Min CHEN ; Zhuo LIN ; Yi CHEN ; Hui-Cong CHEN ;
Chinese Journal of Infectious Diseases 2001;0(05):-
Objective To set up a system in vitro to rapidly recover plasma proteins lost during artificial-liver treatment.Methods The polyprotein precipitation was obtained by all proteins whose isoelectric point pH value were between 7.3 and 5.1,which collided with each other and aggregated using gradient pH co-precipitation(adding 1 mol/L citric acid slowly in the plasma solution to change the pH values gradually from 7.3 to 5.1 in 5 h)combined with salting out(degree of saturation of NaCl is 33%,reacted for 5.5 h at 4℃)or low-temperature ethanol precipitation(40% ethanol, reacted for 5.5 h at -7℃)so that to get rid of toxicants by discarding the supernatant.Results In the range of pH 5.1-7.3,50%(29g/57g)of the total plasma proteins had been recovered by the gradient pH salting out and 41%(25 g/61g)by the gradient pH low-temperature ethanol co-precipi- tation.The protein remained in the supernatant was mostly albumin and its combined bilirubin.The levels of total bilirubin decreased to 0.07% and 0.06% of the original levels by these two methods respectively and the serum HBV DNA level decreased to be undetected(quantitative PCR).Conclu- sions The proteins with close isoelectric point can co-precipitated with the presence of high concen- tration of NaCl or low-temperature ethanol and by changing the pH value gradually.The total protein in the discarded plasma during artificial-liver treatment can be recovered rapidly using the gradient pH coprecipitation.

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