Background::Total human immunodeficiency virus (HIV) DNA and integrated HIV DNA are widely used markers of HIV persistence. Droplet digital polymerase chain reaction (ddPCR) can be used for absolute quantification without needing a standard curve. Here, we developed duplex ddPCR assays to detect and quantify total HIV DNA and integrated HIV DNA.Methods::The limit of detection, dynamic ranges, sensitivity, and reproducibility were evaluated by plasmid constructs containing both the HIV long terminal repeat (LTR) and human CD3 gene (for total HIV DNA) and ACH-2 cells (for integrated HIV DNA). Forty-two cases on stable suppressive antiretroviral therapy (ART) were assayed in total HIV DNA and integrated HIV DNA. Correlation coefficient analysis was performed on the data related to DNA copies and cluster of differentiation 4 positive (CD4 +) T-cell counts, CD8 + T-cell counts and CD4/CD8 T-cell ratio, respectively. The assay linear dynamic range and lower limit of detection (LLOD) were also assessed. Results::The assay could detect the presence of HIV-1 copies 100% at concentrations of 6.3 copies/reaction, and the estimated LLOD of the ddPCR assay was 4.4 HIV DNA copies/reaction (95% confidence intervals [CI]: 3.6-6.5 copies/reaction) with linearity over a 5-log 10-unit range in total HIV DNA assay. For the integrated HIV DNA assay, the LLOD was 8.0 copies/reaction (95% CI: 5.8-16.6 copies/reaction) with linearity over a 3-log 10-unit range. Total HIV DNA in CD4 + T cells was positively associated with integrated HIV DNA ( r = 0.76, P <0.0001). Meanwhile, both total HIV DNA and integrated HIV DNA in CD4 + T cells were inversely correlated with the ratio of CD4/CD8 but positively correlated with the CD8 + T-cell counts. Conclusions::This ddPCR assay can quantify total HIV DNA and integrated HIV DNA efficiently with robustness and sensitivity. It can be readily adapted for measuring HIV DNA with non-B clades, and it could be beneficial for testing in clinical trials.

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.

Objective To investigate the effect and mechanism of Tiaobu Xinshen Prescription(Codonopsis Radix,Polygoni Multiflori Radix Praeparata,Lycii Fructus,Astragali Radix,etc.)on synaptic plasticity in 5xFAD transgenic mice with Alzheimer's disease(AD).Methods Eighteen 5-month-old male wild-type(WT)mice and 18 5xFAD transgenic mice were randomly divided into control group(0.9％NaCl),Tiaobu Xinshen Prescription group(granules,4.18 g·kg-1)and Aricept group(Donepezil Hydrochloride,0.625 mg·kg-1),with six mice in each group.According to the above groups,the rats were given intragastric administration once a day for 60 days.The ultrastructure of hippocampus in mice was observed by transmission electron microscopy.Western Blot was used to detect the protein expression levels of Synaptophsin,PSD-95,p-NMDAR1,NMDAR1,p-CaMKⅡa,CaMKⅡa,PI3K,p-Akt,Akt,p-mTOR and mTOR in mouse cortical tissue.Results Compared with the WT mice control group,the ultrastructure of synapses in the hippocampal CA1 region of the 5xFAD control group was irregular,the mitochondria was atrophied and reduced,the mitochondrial cristae was broken and disappeared,the synaptic membrane was irregularly and curved,the synaptic vesicles were reduced,and the postsynaptic density(PDS)was thinned or even broken.The protein expressions of Synaptophsin,PSD-95,p-NMDAR1/NMDAR1,p-CaMKⅡa/CaMKⅡa,PI3K,p-Akt/Akt and p-mTOR/mTOR in the cortex were significantly down-regulated(P<0.05).Compared with the mice in the 5xFAD control group,the ultrastructure of synapses in the hippocampal CA1 region of the mice in the Tiaobu Xinshen Prescription 5xFAD group was significantly changed,the number of mitochondria was increased,the number of synaptic vesicles was increased,the synaptic membrane was intact,and the postsynaptic density was thickened.The protein expressions of Synaptophsin,PSD-95,p-NMDAR1/NMDAR1,p-CaMKⅡa/CaMKⅡa,PI3K,p-Akt/Akt and p-mTOR/mTOR in the cortex were significantly up-regulated(P<0.05).Conclusion Tiaobu Xinshen Prescription may promote the synthesis of synaptic plasticity-related proteins by activating the PI3K/Akt/mTOR pathway,thereby improving the cognitive dysfunction of AD.

Purpose/Significance To expound the development of the application of new generation intelligent technologies in the medi-cal field in China,and to provide ideas for the future development of intelligent medical technologies in China.Method/Process Literature related to the application of medical intelligence technology from 2000 to 2023 is searched and screened on CNKI,and the knowledge graph is drawn by descriptive statistical analysis and CiteSpace software for visualization analysis.Result/Conclusion The number of papers pub-lished in the application research field of domestic medical intelligence technology shows a trend of steady growth and then leveling off.The hot keywords mainly include smart medical care,health care,health management,etc.,forming 8 main cluster sets.Future research should strengthen cooperation and exchanges between researchers and organizations,and pay more attention to the challenges posed by the applica-tion of technology in terms of security and privacy,technology supervision,talent team construction and technological energy consumption.

Objective:To analyze and summarize the clinical characteristics of mpox patients, thereby to rise clinicians′ awareness of severe mpox and provide a foundation for clinical diagnosis and treatment.Methods:The clinical data were retrospectively analyzed in 41 mpox patients treated at the Beijing You′an Hospital, Capital Medical University, from June to November 2023. Patients were categorized into mild and severe cases based on clinical manifestations, and laboratory results of the two groups were compared. Statistical analysis was performed using the Mann-Whitney U test. Results:The clinical manifestations of 41 mpox patients mainly included fever, rash and lymphadenopathy. Five patients with severe mpox might develop serious complications, including bacterial pneumonia, type Ⅰ respiratory failure, fungal infections, penile or perianal dry gangrene, penile soft tissue edema, intestinal obstruction, septic shock, perianal abscess, and necrotizing fasciitis. Patients with severe mpox had significantly higher white blood cell count (WBC), neutrophil count and C-reactive protein (CRP) level compare to those with mild cases (14.60(9.92, 24.08)×10 9/L vs 6.41(5.64, 8.37)×10 9/L, 12.43(7.02, 21.15)×10 9/L vs 3.35(2.46, 5.03)×10 9/L, 108.20(56.20, 124.10) mg/L vs 16.6(6.25, 49.98) mg/L), while the albumin level and CD4 + T lymphocyte count in the severe group were significantly lower compared to the mild cases (31.80(22.90, 35.15) g/L vs 44.70(42.90, 47.40) g/L, 24.00(12.00, 81.50)/μL vs 606.00(414.50, 767.50)/μL)). All these differences were statistically significant ( U=2.81, 3.02, 2.98, 3.56 and 3.26, respectively, all P<0.01). Conclusions:In clinical practice, clinicians should be vigilant for the possibility of severe mpox if patients exhibit a significant increase of WBC and CRP, a significant decrease in CD4 + T lymphocyte count, or if they develop severe complications.

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.

[Objective]To investigate the impact of varying concentrations of baicalein on the proliferation and biological responses of MC3T3-E1 cells,as well as the antibacterial efficacy of baicalein against prevalent oral bacteria,and to elucidate the underlying mechanisms.[Methods]MC3T3-E1 cells were exposed to different concentrations of baicalein(0,6,12,18,and 24 μmol/L)and cell viability was determined by using the CCK-8 assay.Alkaline phosphatase(ALP)activity of MC3T3-E1 cells following osteogenic induction was assessed.RT-PCR was used to examine the expression of RunX2,BMP2,and Osterix.After 24 hours of treatment,the antibacterial potential of baicalein against Escherichia coli,Staphylococcus Aureus and Streptococcus Sanguis was evaluated by using the K-B paper disk method.[Results]Baicalein exhibited a modest reduction in proliferation of MC3T3-E1 cells but without affecting their sustained proliferation.Baicalein at a concentration of 18 μmol/L enhanced ALP activity of MC3T3-E1 cells,upregulated BMP2 and Osterix expression,downregulated RunX2 expression,significantly inhibited the proliferation of Staphylococcus Aureus and Streptococcus Sanguis(P<0.05).[Conclusions]Baicalein at an optimal concentration(18 μmol/L)demonstrated a promotional effect on the osteogenic differentiation of MC3T3-E1 cells and effectively suppressed the proliferation of common oral bacteria,including Staphylococcus Aureus and Streptococcus Sanguis.

Pulmonary hypertension has a high mortality rate,and although targeted therapy is available,it is still incurable,and the long-term prognosis for patients is poor.As a tyrosine kinase inhibitor,imatinib was approved for marketing in China in 2002 for the treatment of chronic myelogenous leukemia and other tumor diseases.In addition to the antitumor effects,imatinib was found to improve hemodynamics and exercise tolerance in patients with severe pulmonary arterial hypertension,but the safety was suboptimal.With the emergence of new formulations of imatinib targeted at the lungs,it is expected to become a new targeted drug for pulmonary arterial hypertension.

Background::Cluster of differentiation 8 (CD8 T) cells play critical roles in eradicating human immunodeficiency virus (HIV)-1 infection, but little is known about the effects of T cells expressing CD8 at low levels (CD8 low) or high levels (CD8 high) on HIV-1 replication inhibition after HIV-1 invasion into individual. Methods::Nineteen patients who had been acutely infected with HIV-1 (AHI) and 20 patients with chronic infection (CHI) for ≥2 years were enrolled in this study to investigate the dynamics of the quantity, activation, and immune responses of CD3 +CD8 low T cells and their counterpart CD3 +CD8 high T cells at different stages of HIV-1 infection. Results::Compared with healthy donors, CD3 +CD8 low T cells expanded in HIV-1-infected individuals at different stages of infection. As HIV-1 infection progressed, CD3 +CD8 low T cells gradually decreased. Simultaneously, CD3 +CD8 high T cells was significantly reduced in the first month of AHI and then increased gradually as HIV-1 infection progressed. The classical activation of CD3 +CD8 low T cells was highest in the first month of AHI and then reduced as HIV-1 infection progressed and entered the chronic stage. Meanwhile, activated CD38 -HLA-DR +CD8 low T cells did not increase in the first month of AHI, and the number of these cells was inversely associated with viral load ( r = -0.664, P = 0.004) but positively associated with the CD4 T-cell count ( r = 0.586, P = 0.014). Increased programmed cell death protein 1 (PD-1) abundance on CD3 +CD8 low T cells was observed from the 1st month of AHI but did not continue to be enhanced, while a significant T cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibition motif (ITIM) domains (TIGIT) abundance increase was observed in the 12th month of infection. Furthermore, increased PD-1 and TIGIT abundance on CD3 +CD8 low T cells was associated with a low CD4 T-cell count (PD-1: r = -0.456, P = 0.043; TIGIT: r = -0.488, P = 0.029) in CHI. Nonetheless, the nonincrease in PD-1 expression on classically activated CD3 +CD8 low T cells was inversely associated with HIV-1 viremia in the first month of AHI ( r = -0.578, P = 0.015). Notably, in the first month of AHI, few CD3 +CD8 low T cells, but comparable amounts of CD3 +CD8 high T cells, responded to Gag peptides. Then, weaker HIV-1-specific T-cell responses were induced in CD3 +CD8 low T cells than CD3 +CD8 high T cells at the 3rd and 12th months of AHI and in CHI. Conclusions::Our findings suggest that CD3 +CD8 low T cells play an anti-HIV role in the first month of infection due to their abundance but induce a weak HIV-1-specific immune response. Subsequently, CD3 +CD8 low T-cell number decreased gradually as infection persisted, and their anti-HIV functions were inferior to those of CD3 +CD8 high T cells.

Objective:To construct an "Internet + transitional care" scheme for patients with atrial fibrillation after radiofrequency ablation based on ABC overall pathway management, so as to provide reference for the transitional care of patients with atrial fibrillation after surgery.Methods:Through literature review, the first draft of "Internet + transitional care" scheme for patients with atrial fibrillation after radiofrequency ablation based on ABC overall path management was developed. The convenient sampling method was adopted, 16 experts were selected for two rounds of Delphi expert letter consultations and program items were adjusted. Finally, the final draft of "Internet + transitional care" scheme for patients with atrial fibrillation after radiofrequency ablation based on ABC overall pathway management was formed.Results:The effective recovery rates of the two rounds of expert letter consultation were 100.00% (16/16) and 93.75% (15/16) , and the authority coefficients were 0.87 and 0.88, respectively. The Kendall's concordance coefficients of the two rounds of expert consultation were statistically significant ( P<0.05 ) . Finally, a scheme of 70 items was formed, including the day of discharge, 1 week after surgery and 1, 3 and 6 months after surgery. Conclusions:The scheme constructed in this study is scientific and operable and comprehensively reflects the comprehensive management strategy from three aspects, which is worthy of clinical application.