Objective: To develop a simple digital chylous plasma device and validate its ability to accurately, standardly, and non-destructively determine chylous plasma in blood banks and clinical transfusions in hospitals. Methods: A digital chylous plasma assessment device was designed and manufactured. This device was used to measure the chylous degrees of chylous plasma samples before freezing, after freeze-thawing, before viral inactivation, and after viral inactivation. The measured chylosity index values were categorized according to the requirements specified in Appendix A of the Chinese national standard GB 18469-2001 "Quality Requirements for Whole Blood and Blood Components". This process established a digital standard for chylous plasma, enabling the identification of severe, moderate and mild chylous plasma, and non-chylous plasma. Results: The initial simple product of the digital chylous assessment device was successfully designed and manufactured. There was no significant difference in the degree of chylous plasma between pre-freezing 468.11±217.73 lux and post-thawing 538.91±273.39 lux of chylous plasma (P>0.05), or between pre-viral inactivation 858.33±387.79 lux and post-viral inactivation 928.33±166.51 lux of chylous plasma (P>0.05). The median of chylous degree values for plasma chylous index grades 0 to 6 were 45 lux, 250 lux, 620 lux, 835 lux, 1 130 lux, 1 390 lux, and 1 700 lux, respectively. The defined cutoff values/ranges for the chylous degree values corresponding to plasma chylous index grade 0 to 6 were ≤125 lux, 126-465 lux, 466-740 lux, 741-1 000 lux, 1 001-1 233 lux, 1 234-1 560 lux, and ≥1 561 lux. Conclusion: This study successfully developed the initial product of the digital chylous device and established digital standards for classifying chylous plasma. The device demonstrates the potential to meet the needs for assessment of chylous plasma in both blood banks and clinical transfusions in hospitals, thereby promoting the development and application of standardized, non-destructive chylous plasma assessment technology.

BACKGROUND:As the end bearing joint of the human body,the ankle joint bears the top-down pressure of the body,which leads to the ankle joint is easy to be damaged in the movement,can induce functional ankle instability,which negatively affects daily life.The study of lower extremity biomechanics in patients with functional ankle instability during counter movement jump is of great significance for scientific training,prevention of ankle injury,and clinical rehabilitation after injury. OBJECTIVE:To investigate the kinetics and kinematics of lower limbs in the longitudinal jumping of functional ankle instability population. METHODS:From March to September 2023,15 male patients with functional ankle instability and 15 healthy people,aged 22-28 years old,were recruited in Soochow University.All subjects completed counter movement jump experiment.Vicon infrared high-speed motion capture system and Kistler three-dimensional force measuring table were used to simultaneously collect the lower limb kinematics and kinetics indexes of the two groups of subjects at the take-off stage of counter movement jump,the instant off the ground,the initial landing moment and the peak moment of vertical ground reaction force. RESULTS AND CONCLUSION:(1)At the instant off the ground,the affected side of the functional ankle instability group showed smaller knee internal rotation moment(P=0.020)and smaller ankle internal rotation moment(P=0.009)compared with the affected side of the healthy control group.(2)At the moment of landing,the affected side of the functional ankle instability group showed a smaller hip flexion angle than the affected side of the healthy control group(P=0.039).Compared with the healthy control group,functional ankle instability group showed smaller hip abduction angle(P=0.022),smaller knee varus angle(P=0.010),larger knee external rotation angle(P=0.021),smaller ankle varus angle(P=0.004),and smaller external ankle rotation angle(P=0.008).(3)At the peak of vertical ground reaction force,functional ankle instability group showed a smaller ankle varus angle than healthy control group(P=0.044).(4)The results showed that the lower limb biomechanical characteristics of the patients with functional ankle instability were abnormal compared with the healthy people during counter movement jump,which mainly showed the changes of the kinematics and kinetics indexes of the lower limb joints in the sagittal plane and the frontal plane at the moment of lift-off and landing.These changes reflect that people with functional ankle instability adopt rigid take-off and landing patterns when performing counter movement jump,tend to transfer the load of the affected ankle joint to other joints of the lower limb,and show compensatory phenomenon of the healthy lower limb.Therefore,detection and correction of abnormal biomechanical features should be a part of rehabilitation training for those with functional ankle instability.

Objective:To understand the etiological surveillance and drug resistance characteristics of Legionella pneumophila (LP) from the aqueous environment of public places in Shanghai, from 2011 to 2020, and provide evidence for surveillance of the disease. Methods:Environmental water samples were systematically collected from public venues in urban and suburban districts of Shanghai for LP surveillance. All the identified LP isolates underwent a series of testings including serotyping, pulsed field gel electrophoresis (PFGE), sequence-based typing, and antimicrobial susceptibility testing. χ2 test or Cochran-Armitage trend tests were used for statistical analysis and for temporal resistance patterns. Results:Among 6 263 water samples, the LP-positive rate was 20.93% (1 311/6 263). The positivity rate decreased from 24.98% (287/1 149) in 2011-2012 to 20.02% (1 024/5 114) in 2013-2020 ( χ2=13.92, P<0.001), with the highest monthly positivity observed from June to August (23.79%, 745/3 132). A total of 1 365 LP strains were isolated, of which 912 were further characterized, including 10 serotypes, 149 PFGE patterns, and 33 sequence types (ST). The predominant serotype was Lp1 (86.84%, 792/912), and the dominant ST was ST752 (29.50%, 269/912). ST clustering revealed two major clonal groups CG1 and CG2, accounting for 91.12% (831/912) of the isolates. The 190 LPs involved in the drug sensitivity test showed three resistance profiles: azithromycin resistance (31.05%, 59/190), ciprofloxacin resistance (0.53%, 1/190) and azithromycin+ciprofloxacin resistance (0.53%, 1/190). Azithromycin-resistant strains were predominantly ST1 (64.41%, 38/59). The antimicrobial resistance rate showed a significant decline, from 48.65% (18/37) in 2011-2012 to 28.10% (43/153) in 2013-2020 ( χ2=9.38, P=0.002). Conclusions:Compared to from 2011 to 2012, both the positivity rate and antimicrobial resistance prevalence of LP in public aqueous environments of Shanghai exhibited an overall decline from 2013 to 2020. The predominant types of LP were serotype Lp1 and sequence type ST752, with notable high-level resistance to azithromycin. Measures as enhancing the enforcement of water safety regulations and prioritizing surveillance of azithromycin resistance in LP were recommended to mitigate public health risks.

OBJECTIVE To observe the effect of electroacupuncture(EA)preconditioning at Tianshu(ST25)and Neiguan(PC6)on cardiac function and UCP1/BMP3b signaling pathway in brown adipose tissue(BAT)in mice with acute myocardial infarc-tion(AMI),so as to explore the potential mechanism of EA at different acupoints in improving myocardial infarction.METHODS Healthy adult mice and BAT excision mice were selected as the research objects,and they were divided into sham operation group,model group,PC6 group,and ST25 group.After one week of adaptive feeding,the mice in the intervention group were pretreated with bilateral EA at PC6 or ST25 for 20 minutes,respectively,and the AMI model was established by ligating the left anterior descending branch of the coronary artery.In the BAT resection group,BAT resection of the scapular region was performed before EA,and the rest of the intervention remained the same as before.Echocardiography was used to detect the changes in cardiac function.TTC staining was used to observe the myocardial infarct size.ELISA was used to detect the serum levels of cTnT and BMP3b in each group.The qPCR was used to detect the relative expression of β3-AR,UCP1 and BMP3b mRNA in mouse BAT.The protein expression of BMP3b in BAT and p-Smad1/5 in the heart were detected by Western blot.RESULTS Compared with the sham operation group,the left ven-tricular EF and FS of the model group mice were decreased(P<0.001),the white infarct area was increased(P<0.001),the cTnT level in serum was increased(P<0.001),the mRNA expression levels of β3-AR,UCP1,and BMP3b in BAT were increased(P<0.05,P<0.01),and the protein expression of BMP3b was increased(P<0.01).The BMP3b content in serum was increased(P<0.001),and the protein expression of p-Smad1/5 in the heart was increased(P<0.01).Compared with the model group,the left ventricular EF and FS of mice in the ST25 and PC6 group were increased(P<0.001),the area of white infarction was reduced(P<0.001),and the cTnT level in serum was decreased(P<0.05,P<0.01).The mRNA contents of β3-AR,UCP1,and BMP3b in BAT of the ST25 group were significantly increased(P<0.05,P<0.01,P<0.001),the protein expression of BMP3b was increased(P<0.01),and the protein expression of p-Smad1/5 in the heart was increased(P<0.01),while there was no significant change in the PC6 group.After BAT resection,compared with the model group,the left ventricular EF and FS of the mice in the PC6 group were increased(P<0.001),the area of white infarction was reduced(P<0.001),and the cTnT in serum was decreased(P<0.001),while there was no significant change in the ST25 group;there was no significant change in the protein expression of p-Smad1/5 in the heart and the BMP3b content in the serum of the PC6 and ST25 groups.CONCLUSION EA pretreatment at either ST25 or PC6 acupoints can produce myocardial protective effects.The protective effect of ST25 may be through influencing the UCP1/BMP3b signaling path-way in BAT,while PC6 does not depend on this pathway.

Objective To investigate the distribution characteristics of Klebsiella pneumoniae(KP),hyperviru-lent Klebsiella pneumoniae(hvKP),carbapenem-resistant Klebsiella pneumoniae(CRKP),and hypervirulent car-bapenem-resistant Klebsiella pneumoniae(hv-CRKP/CR-hvKP)in the environment of general intensive care unit(ICU)at medical institutions,and provide reference for environment assessment as well as healthcare-associated in-fection(HAI)prevention and control in ICU.Methods A total of 3 336 environmental specimens were collected from general ICUs of medical institutions in Shanghai in 2019 and 2023.After strain isolation,antimicrobial suscep-tibility testing and whole genome sequencing were conducted.Results The detection rate of KP was 1.59％(n=53),among which hvKP,CRKP,and hv-CRKP/CR-hvKP accounted for 37.74％(20/53),52.83％(28/53),and 24.53％(13/53)of the total detected strains,respectively.The main types of hvKP were ST11-KL64 and ST11-KL25,CRKP were ST15-KL19 and ST11-KL25,hv-CRKP/CR-hvKP were ST11-KL25 and ST11-KL64.The main carried resistance genes included fosA,oqx AB,tet(A),blaTEM-1B,blaKPC-2,qnrS11,etc.All strains carried viru-lence genes fimH,iutA,ent A,entB,entC,entD,entE,and entF,with only one strain carrying rmp A gene.Conclusion KP contamination is widespread in general ICU environment of medical institutions,predominantly ST11 and ST15,presenting a polymorphic distribution.CRKP and hvKP account for a relatively high proportion,and multidrug resistance is serious.Co-evolution of drug resistance and virulence presents in KP,and poses signifi-cant infection and pathogenic risks to patients,necessitating enhanced clinical vigilance and preparedness for poten-tial outbreaks.

Objective:To understand the etiological surveillance and drug resistance characteristics of Legionella pneumophila (LP) from the aqueous environment of public places in Shanghai, from 2011 to 2020, and provide evidence for surveillance of the disease. Methods:Environmental water samples were systematically collected from public venues in urban and suburban districts of Shanghai for LP surveillance. All the identified LP isolates underwent a series of testings including serotyping, pulsed field gel electrophoresis (PFGE), sequence-based typing, and antimicrobial susceptibility testing. χ2 test or Cochran-Armitage trend tests were used for statistical analysis and for temporal resistance patterns. Results:Among 6 263 water samples, the LP-positive rate was 20.93% (1 311/6 263). The positivity rate decreased from 24.98% (287/1 149) in 2011-2012 to 20.02% (1 024/5 114) in 2013-2020 ( χ2=13.92, P<0.001), with the highest monthly positivity observed from June to August (23.79%, 745/3 132). A total of 1 365 LP strains were isolated, of which 912 were further characterized, including 10 serotypes, 149 PFGE patterns, and 33 sequence types (ST). The predominant serotype was Lp1 (86.84%, 792/912), and the dominant ST was ST752 (29.50%, 269/912). ST clustering revealed two major clonal groups CG1 and CG2, accounting for 91.12% (831/912) of the isolates. The 190 LPs involved in the drug sensitivity test showed three resistance profiles: azithromycin resistance (31.05%, 59/190), ciprofloxacin resistance (0.53%, 1/190) and azithromycin+ciprofloxacin resistance (0.53%, 1/190). Azithromycin-resistant strains were predominantly ST1 (64.41%, 38/59). The antimicrobial resistance rate showed a significant decline, from 48.65% (18/37) in 2011-2012 to 28.10% (43/153) in 2013-2020 ( χ2=9.38, P=0.002). Conclusions:Compared to from 2011 to 2012, both the positivity rate and antimicrobial resistance prevalence of LP in public aqueous environments of Shanghai exhibited an overall decline from 2013 to 2020. The predominant types of LP were serotype Lp1 and sequence type ST752, with notable high-level resistance to azithromycin. Measures as enhancing the enforcement of water safety regulations and prioritizing surveillance of azithromycin resistance in LP were recommended to mitigate public health risks.

OBJECTIVE To observe the effect of electroacupuncture(EA)preconditioning at Tianshu(ST25)and Neiguan(PC6)on cardiac function and UCP1/BMP3b signaling pathway in brown adipose tissue(BAT)in mice with acute myocardial infarc-tion(AMI),so as to explore the potential mechanism of EA at different acupoints in improving myocardial infarction.METHODS Healthy adult mice and BAT excision mice were selected as the research objects,and they were divided into sham operation group,model group,PC6 group,and ST25 group.After one week of adaptive feeding,the mice in the intervention group were pretreated with bilateral EA at PC6 or ST25 for 20 minutes,respectively,and the AMI model was established by ligating the left anterior descending branch of the coronary artery.In the BAT resection group,BAT resection of the scapular region was performed before EA,and the rest of the intervention remained the same as before.Echocardiography was used to detect the changes in cardiac function.TTC staining was used to observe the myocardial infarct size.ELISA was used to detect the serum levels of cTnT and BMP3b in each group.The qPCR was used to detect the relative expression of β3-AR,UCP1 and BMP3b mRNA in mouse BAT.The protein expression of BMP3b in BAT and p-Smad1/5 in the heart were detected by Western blot.RESULTS Compared with the sham operation group,the left ven-tricular EF and FS of the model group mice were decreased(P<0.001),the white infarct area was increased(P<0.001),the cTnT level in serum was increased(P<0.001),the mRNA expression levels of β3-AR,UCP1,and BMP3b in BAT were increased(P<0.05,P<0.01),and the protein expression of BMP3b was increased(P<0.01).The BMP3b content in serum was increased(P<0.001),and the protein expression of p-Smad1/5 in the heart was increased(P<0.01).Compared with the model group,the left ventricular EF and FS of mice in the ST25 and PC6 group were increased(P<0.001),the area of white infarction was reduced(P<0.001),and the cTnT level in serum was decreased(P<0.05,P<0.01).The mRNA contents of β3-AR,UCP1,and BMP3b in BAT of the ST25 group were significantly increased(P<0.05,P<0.01,P<0.001),the protein expression of BMP3b was increased(P<0.01),and the protein expression of p-Smad1/5 in the heart was increased(P<0.01),while there was no significant change in the PC6 group.After BAT resection,compared with the model group,the left ventricular EF and FS of the mice in the PC6 group were increased(P<0.001),the area of white infarction was reduced(P<0.001),and the cTnT in serum was decreased(P<0.001),while there was no significant change in the ST25 group;there was no significant change in the protein expression of p-Smad1/5 in the heart and the BMP3b content in the serum of the PC6 and ST25 groups.CONCLUSION EA pretreatment at either ST25 or PC6 acupoints can produce myocardial protective effects.The protective effect of ST25 may be through influencing the UCP1/BMP3b signaling path-way in BAT,while PC6 does not depend on this pathway.

Objective To investigate the distribution characteristics of Klebsiella pneumoniae(KP),hyperviru-lent Klebsiella pneumoniae(hvKP),carbapenem-resistant Klebsiella pneumoniae(CRKP),and hypervirulent car-bapenem-resistant Klebsiella pneumoniae(hv-CRKP/CR-hvKP)in the environment of general intensive care unit(ICU)at medical institutions,and provide reference for environment assessment as well as healthcare-associated in-fection(HAI)prevention and control in ICU.Methods A total of 3 336 environmental specimens were collected from general ICUs of medical institutions in Shanghai in 2019 and 2023.After strain isolation,antimicrobial suscep-tibility testing and whole genome sequencing were conducted.Results The detection rate of KP was 1.59％(n=53),among which hvKP,CRKP,and hv-CRKP/CR-hvKP accounted for 37.74％(20/53),52.83％(28/53),and 24.53％(13/53)of the total detected strains,respectively.The main types of hvKP were ST11-KL64 and ST11-KL25,CRKP were ST15-KL19 and ST11-KL25,hv-CRKP/CR-hvKP were ST11-KL25 and ST11-KL64.The main carried resistance genes included fosA,oqx AB,tet(A),blaTEM-1B,blaKPC-2,qnrS11,etc.All strains carried viru-lence genes fimH,iutA,ent A,entB,entC,entD,entE,and entF,with only one strain carrying rmp A gene.Conclusion KP contamination is widespread in general ICU environment of medical institutions,predominantly ST11 and ST15,presenting a polymorphic distribution.CRKP and hvKP account for a relatively high proportion,and multidrug resistance is serious.Co-evolution of drug resistance and virulence presents in KP,and poses signifi-cant infection and pathogenic risks to patients,necessitating enhanced clinical vigilance and preparedness for poten-tial outbreaks.

DNMT3A encodes a DNA methyltransferase involved in development, cell differentiation, and gene transcription, which is mutated and aberrant-expressed in cancers. Here, we revealed that loss of DNMT3A promotes malignant phenotypes in lung cancer. Based on the epigenetic inhibitor library synthetic lethal screening, we found that small-molecule HDAC6 inhibitors selectively killed DNMT3A-defective NSCLC cells. Knockdown of HDAC6 by siRNAs reduced cell growth and induced apoptosis in DNMT3A-defective NSCLC cells. However, sensitive cells became resistant when DNMT3A was rescued. Furthermore, the selectivity to HDAC6 inhibition was recapitulated in mice, where an HDAC6 inhibitor retarded tumor growth established from DNMT3A-defective but not DNMT3A parental NSCLC cells. Mechanistically, DNMT3A loss resulted in the upregulation of HDAC6 through decreasing its promoter CpG methylation and enhancing transcription factor RUNX1 binding. Notably, our results indicated that HIF-1 pathway was activated in DNMT3A-defective cells whereas inactivated by HDAC6 inhibition. Knockout of HIF-1 contributed to the elimination of synthetic lethality between DNMT3A and HDAC6. Interestingly, HIF-1 pathway inhibitors could mimic the selective efficacy of HDAC6 inhibition in DNMT3A-defective cells. These results demonstrated HDAC6 as a HIF-1-dependent vulnerability of DNMT3A-defective cancers. Together, our findings identify HDAC6 as a potential HIF-1-dependent therapeutic target for the treatment of DNMT3A-defective cancers like NSCLC.

BACKGROUND:The clinical application of Mongolian Medicine Erdun-uril in the treatment of osteoarthritis is effective,but its therapeutic effect and related mechanism are still unclear. OBJECTIVE:To observe the repairing process of osteoarthritis in rats by Mongolian Medicine Erdun-uril and analyze its mechanism. METHODS:Thirty male Sprague-Dawley rats(8 weeks of age)were selected to establsih an osteoarthritis model using intraarticular injection of sodium iodoacetate solution into the unilateral knee joint.Two weeks after modeling,the model rats were randomly divided into three groups:control group(n=6)was given normal saline by gavage,and low and high dose groups(n=12 per group)were given Erdun-uril 1.4 and 2 g,respectively,3 times per day.After 2 and 4 weeks of continuous administration,blood samples,articular cartilage and periarticular bone tissue were taken from rats to detect changes in cartilage metabolic markers(cartilage oligomeric matrix protein,proteoglycan),bone metabolic markers(bone alkaline phosphatase,keratan sulfate),inflammatory markers(interleukin 6,tumor necrosis factor α),lipid metabolic markers(total cholesterol,triacylglycerol).Histopathological changes were also observed. RESULTS AND CONCLUSION:qRT-PCR results showed that after 2 and 4 weeks of administration,the mRNA expression of cartilage oligomeric matrix protein,proteoglycan and bone alkaline phosphatase and keratan sulfate in knee cartilage of rats were significantly reduced in the low and high dose groups compared with the control group(P<0.05,P<0.01,P<0.001),with the reduction being more significant in the high dose group.After 2 and 4 weeks of administration,compared with the control group,serum tumor necrosis factor α and interleukin 6 levels were decreased(P<0.01,P<0.001)and total cholesterol and triacylglycerol levels were increased(P<0.05,P<0.01,P<0.001)in the low and high dose groups,with the improvement being more significant in the high dose group.Hematoxylin-eosin staining results showed that in the control group,there were large defective areas on the surface of the knee cartilage with unknown structure and severe inflammatory cell infiltration;in the low dose group,the surface of the knee cartilage was rough with disordered cell arrangement and the cartilage structure did not improve significantly after 2 weeks of administration,while the number of inflammatory cells in the knee cartilage was reduced and the cartilage structure was restored to some extent after 4 weeks of administration;in the high dose group,the structure of the knee cartilage become clear after 2 weeks of administration and even clearer after 4 weeks of administration,and the inflammation was significantly reduced after 2 weeks of administration.To conclude,Mongolian Medicine Erdun-uril with better anti-inflammatory properties can improve cartilage metabolism and obviously alleviate osteoarthritis in rats.