OBJECTIVE To establish a method for simultaneous determination of venetoclax， busulfan and voriconazole in plasma of patients with acute myeloid leukemia （AML）， and apply it clinically. METHODS Plasma samples were subjected to protein precipitation using acetonitrile and subsequently analyzed by liquid chromatography-tandem mass spectrometry （LC-MS/MS） using venetoclax-D 8 ， busulfan-D 8 and posaconazole as internal standards. The separation was performed on a Phenomenex Kinetex ® C 18 column with a mobile phase composed of 0.1% formic acid solution （2 mmol/L ammonium acetate）-0.1% formic acid in acetonitrile （gradient elution） at a flow rate of 0.8 mL/min. The column temperature was set at 40 ℃， the sample size was 5 μL， and the total run time was 3.10 min. An electrospray ionization source was employed， and positive ion scanning was conducted using multiple reaction monitoring mode. The ion pairs used for quantitative analysis included m/z 868.4→636.3 （venetoclax）， m/z 264.1→151.1 （busulfan）， and m/z 350.1→224.0 （voriconazole）. The above LC-MS/MS method was adopted to determine plasma concentrations of venetoclax and voriconazole in 10 AML patients， as well as plasma concentration of busulfan in 5 patients undergoing conditioning treatment for allogeneic hematopoietic stem cell transplantation. RESULTS The linear ranges of venetoclax， busulfan and voriconazole were 50-10 000， 15-3 000 and 50-10 000 ng/mL， respectively （ R 2 ≥0.999 0）， with lower limits of quantification of 50， 15 and 50 ng/mL， respectively. The RSDs of intra-day and inter-day precision tests for all three analytes were all less than 10%， with accuracy （relative errors） ranging from －10.00% to 12.96%. The average extraction recovery ranged from 92.54% to 100.95%， and the average matrix effect was 89.98%-101.49%. Dilution reliability covered all dilution factors used in the test samples， and the absolute values of relative errors in stability tests were all≤16.25%. The plasma concentrations of venetoclax， busulfan and voriconazole in enrolled patients were 496.20-4 250.45， 233.48-2 002.28 and 475.51-5 710.18 ng/mL， respectively. CONCLUSIONS The LC-MS/MS method established in this study is rapid， sensitive and easy to operate， and can be used for the therapeutic drug monitoring of venetoclax， busulfan and voriconazole.

Objective: To investigate the mechanism by which serum amyloid P component (SAP) alleviates periodontitis in mice, providing an experimental basis to establish SAP as a novel therapeutic agent for periodontitis. Methods: Ethical approval was obtained from the Institutional Animal Ethics Committee. Periodontitis models were established in wild-type (WT) mice and SAP-transgenic (SAP-Tg) mice, divided into four groups: WT control (WT group), WT periodontitis (WT+P group), SAP-Tg control (Tg group), and SAP-Tg periodontitis (Tg+P group). On day 7, the mice were euthanized, and periodontal tissues, teeth, and alveolar bone were collected. SAP protein expression was detected by enzyme-linked immunosorbent assay (ELISA). Micro-CT and HE staining were used to measure alveolar bone resorption (distance from the cementoenamel junction to the alveolar bone crest). Tartrate-resistant acid phosphatase (TRAP) staining was performed to assess osteoclast number, and immunohistochemistry (IHC) was employed to evaluate macrophage infiltration. The expression levels of inflammatory cytokines including interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were measured by qRT-PCR. Oral microorganism composition was analyzed using 16S ribosomal RNA (16S rRNA) gene sequencing. Additionally, macrophages from WT and SAP-Tg mice were isolated to establish an in vitro inflammation model, divided into WT+LPS and Tg+LPS groups. The expression of macrophage polarization-related genes including inducible nitric oxide synthase (iNOS), CD86, CD163, and CD206) were assessed by qRT-PCR. After the induction of osteoclast differentiation, TRAP staining was performed. Results: ELISA results demonstrated that periodontal tissues from Tg+P group mice exhibited higher levels of SAP expression compared to the WT+P group. Micro-CT and HE staining analyses revealed that the Tg+P group showed reduced alveolar bone resorption, indicated by a shorter distance between the cementoenamel junction and alveolar bone crest, compared to the WT+P group. Furthermore, TRAP staining results indicated a decrease in osteoclast numbers in the Tg+P group compared to the WT+P group. IHC and qRT-PCR results indicated reduced macrophage infiltration and decreased expression of IL-1β, IL-6, and TNF-α in the Tg+P group. Oral microorganism sequencing showed no significant difference in periodontitis-associated pathogenic bacteria between WT+P and Tg+P groups. In vitro experiments demonstrated that compared to the WT+LPS group, the Tg+LPS group exhibited downregulated M1 macrophage markers (iNOS and CD86) and upregulated M2 macrophage markers (CD163 and CD206). TRAP staining confirmed fewer osteoclasts in the Tg+LPS group. Conclusion SAP overexpression effectively alleviates periodontitis severity in mice by inhibiting M1 macrophage polarization, reducing pro-inflammatory cytokine expression, and suppressing osteoclast differentiation, thereby attenuating alveolar bone resorption.

This paper analyzed the traditional Chinese medicine （TCM） and western medical understanding of coronary microvascular disease （CMVD） from the three dimensions of "disease-syndrome-symptom". In western medicine， by summarizing the suspected diagnosis and understanding of CMVD， it is believed that inflammatory responses and vascular endothelial damage are the key mechanisms of the pathogenesis. From the perspective of TCM， the disease location is at blood， vessels and heart， and the fundamental cause is spleen and kidney depletion， closely realted to phlegm， stasis， toxin， wind and qi. Integrating the understanding of both TCM and western medicine， clinical treatment advocates taking the CMVD pathology as the base， and the TCM understanding of pathogenesis as the main focus. The properties of Chinese herbal medicinals is used as the guidance for medication， and the pharmacological understanding as the assisstance of treatment， with the medical history and the severity of the condition are additionally considered. It is finally proposed that during the acute phase， the methods of nourishing yin and resolving toxins， softening hardness and dissipating masses， dispelling wind and unblocking collaterals should be applied to alleviate the emergency. In the subacute phase， the focus should be on raising and lifting qi promote its movement， with flexible use of medicinals that can unblock yang. In the remission phase， the method of tonifying spleen and fortifying kidney should be used to maintain the stability of the condition.

Objective:To investigate the application value of self-pulling latter transection(SPLT)in esophagojeju-nal anastomosis during total laparoscopic total gastrectomy.Methods:From January 2021 to December 2023,80 pa-tients who underwent total laparoscopic total gastrectomy at the Department of General Surgery,Yancheng Third People's Hospital,were selected.Patients were grouped based on the intraoperative esophagojejunal anastomosis method:the SPLT group(35 cases,using SPLT esophagojejunostomy)and the Overlap group(45 cases,using esoph-agojejunal overlapping side-to-side anastomosis).Surgical related indicators,complication incidence,nutritional status,and quality of life assessment were compared between the two groups.Results:The time to first postoperative anal exhaust,the number of lymph nodes dissected,and the postoperative hospital stay were compared between the two groups(P＞0.05).The SPLT group had shorter operation time and esophagojejunal anastomosis time,and less intraopera-tive blood loss compared to the control group(P＜0.05).The incidence of postoperative complications in the SPLT group was 8.57%,and in the Overlap group was 11.11%,with no significant difference between the two groups(P＞0.05).The nutritional status at 6 months postoperatively in the SPLT group[Hb(116.97±10.94)g/L、ALB(39.74±1.29)g/L、PA(0.30±0.45)g/L]and Overlap group[Hb(119.78±12.84)g/L、ALB(39.64±1.42)g/L、PA(0.30±0.36)g/L]were compared(P＞0.05).The quality of life assessment(overall health status,functional domains,symptom domains)at 6 months postoperatively between the SPLT group and the Overlap group was compared(P＞0.05).Conclusion:The use of SPLT esophagojeju-nal anastomosis in total laparoscopic total gastrectomy can improve the surgical efficiency,achieve high anastomosis,and its medium and short-term safety is comparable to that of esophagojejunal overlap side-to-side anastomosis.

Objective:To observe the effect of combining low-frequency transcranial magnetic stimulation (rTMS) with melodic intonation therapy (MIT) in the treatment of aphasia soon after a stroke.Methods:Sixty survivors of a recent stroke with aphasia were randomly divided into a control group, an MIT group and an observation group, each of 20. All received conventional rehabilitation therapy, but for 4 weeks the MIT group was additionally given MIT while the observation group received MIT plus 1Hz rTMS applied over the right inferior frontal gyrus and the posterior superior temporal gyrus. The intensity was 80% of the resting motor threshold. Before and after the treatment, language function and communication ability were evaluated using the Chinese version of the Western Aphasia Battery (WAB) and Communicative Activities of Daily Living (CADL) scale.Results:Significant improvements in the WAB and CADL scores and aphasia quotient (AQ) were observed in both the MIT group and the observation group after the treatment compared to their pre-treatment levels, but the observation group demonstrated significantly higher scores than the other two groups on all three assessments. The MIT group′s average scores were superior to those of the control group.Conclusions:Applying rTMS and MIT in combination significantly improves the speech and the daily communication abilities of aphasics early post-stroke, reducing the severity of their aphasia. The therapy exhibits considerable promise for clinical implementation.

Aim To explore the mechanism of baicalin combined with heat stimulation in treating acute lym-phoblastic leukemia(ALL)based on network pharma-cology and in vitro experiments.Methods The CCK-8 assay was used to screen the suitable conditions for heat stimulation to interfere ALL cell lines Jurkat,CCRF-CEM,Hut-78 and a normal lymphocyte HMy2.CIR,and the effects of baicalin combined with heat stimulation on the proliferation of three ALL cell lines and a normal lymphocyte were tested.The key targets of baicalin combined with fever stimulation for the treatment of ALL were obtained based on network phar-macological analysis,and the potential mechanisms were predicted by gene ontology(GO)annotation and kyoto encyclopedia of genes and genomes(KEGG)en-richment.The expression levels of TNF-α,AKT1,TYMS and CASP3 mRNA in ALL cell lines Jurkat and CCRF-CEM were examined by RT-qPCR with baicalin alone and baicalin combined with heat stimulation.Results The optimal conditions for heat stimulation to intervene ALL cells were 41 ℃ for 24 h,and heat stimulation combined with baicalin synergistically inhibited the growth of ALL cell lines and effectively reduced the cy-totoxicity of baicalin.Based on the network pharmaco-logical analysis,55 intersecting targets of baicalin with ALL diseases and 77 intersecting targets of baicalin with fever were obtained.The results of GO annotation and KEGG enrichment suggested that baicalin com-bined with fever stimulation to intervene ALL might be associated with influencing intracellular reactive oxygen species metabolism,DNA transcription and apoptotic processes involved in cysteine enzymes.Apoptosis,TNF and IL-17 signaling pathways were the key pathways for baicalin combined with heat stimulation in treating ALL.Under heat stimulation at 41 ℃ using SDHA gene as housekeeping gene,in vitro experiments showed that baicalin significantly up-regulated the expression of TNF-α and CASP3,and down-regulated the expression of TYMS in ALL cells.Conclusions Based on net-work pharmacologic analyses and in vitro experiments,baicalin combined with heat stimulation can regulate TNF-α and CASP3 gene levels in ALL cells and de-stroy cellular structure to promote cell apoptosis,thus synergistically treating ALL.

Objective:To investigate the changes in thyroid hormone levels and inflammatory markers in critically ill children，analyze their correlation with disease severity，and explore their potential impact on prognosis,providing references for clinical management and prognosis assessment in critical illness.Methods:A retrospective cohort study was conducted involving 394 pediatric patients admitted to the ICU of the Capital Pediatric Institute Affiliated Children's Hospital from 2019 to 2023.Based on the pediatric critical illness score,patients were divided into three groups:the extremely critical group (score ≤ 70, n=81),the critical group (score 71–80, n=150),and the non-critical group (score>80, n=163).Data collected included thyroid function indicators，inflammatory markers[C-reactive protein（CRP)，procalcitonin(PCT),tumor necrosis factor(TNF)-α，interleukin (IL)，etc.]，clinical information，and outcomes.The correlation between thyroid function indicators and inflammatory markers were analyzed.The predictive value of thyroid function indicators and inflammatory markers for prognosis in critically ill pediatric patients was assessed. Results:Of the 394 children,non-thyroidal disease syndrome occurred in 321 cases,with an overall incidence of 81.5%,which increased with disease severity.Thyroid hormone [total triiodothyronine (TT3),free triiodothyronine (FT3),and total tetraiodothyronine (TT4)] levels were significantly lower in the extremely critical group than in the other groups ( P<0.05).Inflammatory markers such as CRP,PCT,TNF-α,IL-6,IL-8,and IL-10 were significantly higher in the extremely critical group than in the other groups ( P<0.05).Thyroid hormones were negatively correlated with inflammatory markers,and the receivor operating characteristic curves analysis indicated that TT3,FT3,IL-6 and IL-8 levels,could effectively differentiate disease prognosis.Univariate regression model showed significant associations between TT3,FT3,TT4,PCT,IL-8,and IL-10 and disease prognosis.The multivariate Logistic regression model showed IL-6 and IL-8 were independent predictors of disease prognosis. Conclusion:Significant reductions in thyroid hormone levels are closely related to disease severity and poor prognosis.Changes in inflammatory markers reflect the inflammatory state and severity of the disease and impact prognosis.Monitoring thyroid function and inflammatory status is important in clinical management，which provids new insights into prognosis assessment and treatment strategies for critically ill children.

Objective To investigate the antimicrobial resistance of clinical isolates from elderly patients(≥65 years)in major medical institutions across China.Methods Bacterial strains were isolated from elderly patients in 52 hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program during the period from 2015 to 2021.Antimicrobial susceptibility test was carried out by disk diffusion method and automated systems according to the same CHINET protocol.The data were interpreted in accordance with the breakpoints recommended by the Clinical and Laboratory Standards Institute(CLSI)in 2021.Results A total of 514 715 nonduplicate clinical isolates were collected from elderly patients in 52 hospitals from January 1,2015 to December 31,2021.The number of isolates accounted for 34.3％of the total number of clinical isolates from all patients.Overall,21.8％of the 514 715 strains were gram-positive bacteria,and 78.2％were gram-negative bacteria.Majority(90.9％)of the strains were isolated from inpatients.About 42.9％of the strains were isolated from respiratory specimens,and 22.9％were isolated from urine.More than half(60.7％)of the strains were isolated from male patients,and 39.3％isolated from females.About 51.1％of the strains were isolated from patients aged 65-＜75 years.The prevalence of methicillin-resistant strains(MRSA)was 38.8％in 32 190 strains of Staphylococcus aureus.No vancomycin-or linezolid-resistant strains were found.The resistance rate of E.faecalis to most antibiotics was significantly lower than that of Enterococcus faecium,but a few vancomycin-resistant strains(0.2％,1.5％)and linezolid-resistant strains(3.4％,0.3％)were found in E.faecalis and E.faecium.The prevalence of penicillin-susceptible S.pneumoniae(PSSP),penicillin-intermediate S.pneumoniae(PISP),and penicillin-resistant S.pneumoniae(PRSP)was 94.3％,4.0％,and 1.7％in nonmeningitis S.pneumoniae isolates.The resistance rates of Klebsiella spp.(Klebsiella pneumoniae 93.2％)to imipenem and meropenem were 20.9％and 22.3％,respectively.Other Enterobacterales species were highly sensitive to carbapenem antibiotics.Only 1.7％-7.8％of other Enterobacterales strains were resistant to carbapenems.The resistance rates of Acinetobacter spp.(Acinetobacter baumannii 90.6％)to imipenem and meropenem were 68.4％and 70.6％respectively,while 28.5％and 24.3％of P.aeruginosa strains were resistant to imipenem and meropenem,respectively.Conclusions The number of clinical isolates from elderly patients is increasing year by year,especially in the 65-＜75 age group.Respiratory tract isolates were more prevalent in male elderly patients,and urinary tract isolates were more prevalent in female elderly patients.Klebsiella isolates were increasingly resistant to multiple antimicrobial agents,especially carbapenems.Antimicrobial resistance surveillance is helpful for accurate empirical antimicrobial therapy in elderly patients.

BACKGROUND:Hemivertebra deformity should be treated surgically at an early age,but the risk factors for progression of deformity after hemivertebral resection have not been established.OBJECTIVE:To investigate the curative effect of one-stage posterior hemivertebrae resection and pedicle screw fixation in the treatment of congenital scoliosis,and to further explore the risk factors causing the progression of postoperative deformity.METHODS:The medical records of patients who underwent pedicle screw-rod fixation for unilateral hemivertebral deformity from January 2012 to February 2020 and were followed up for at least 2 years were retrospectively analyzed,and a total of 116 patients met the inclusion criteria.All patients were treated with standing anterior and lateral spinal radiographs taken before,after and at each follow-up time point.The segment Cobb angle,the total scoliosis Cobb angle,the proximal complementary Cobb angle,the distal complementary Cobb angle,and the coronal balance distance,apical vertebra distance,upper instrumented vertebra tilt,upper instrumented vertebra disc angle,lower instrumented vertebra tilt,lower instrumented vertebra disc angle,segmental kyphosis/lordosis,thoracic kyphosis,lumbar lordosis and sagittal vertical axis were measured.The progression of deformity and complications were also recorded.RESULTS AND CONCLUSION:(1)Segment Cobb,total scoliosis Cobb,segmental kyphosis,proximal complementary Cobb,and distal complementary Cobb were significantly corrected after operation(P＜0.05),and remained corrected at the last follow-up.Thoracic kyphosis,lumbar lordosis,coronal balance distance,and sagittal vertical axis were all in the normal range pre-operation,after operation and at the last follow-up.(2)During follow-up,10 patients developed coronary decompensation,which was characterized by abnormal progression.(3)Independent sample t-test showed that preoperative total scoliosis Cobb,preoperative apical vertebra distance,age,Risser sign,postoperative upper instrument vertebra tilt and postoperative lowest instrumented vertebra tilt were correlated with postoperative malformation progression(P＜0.05).(4)Multivariate Logistic regression analysis showed that postoperative lowest instrumented vertebra tilt was an independent risk factor for postoperative malformation progression(P=0.002,OR=1.526).(5)Receiver operating characteristic curve analysis showed that a postoperative lowest instrumented vertebra tilt of 8.14° was the optimal threshold for deformity progression after hemivertebrae resection and pedicle rod fixation(sensitivity 0.900,specificity 0.906,area under curve:0.926).(6)It is indicated that the treatment of congenital scoliosis with one-stage posterior hemivertebrae resection and pedicle nail fixation can achieve satisfactory orthopedic effect.Postoperative lowest instrumented vertebra tilt greater than 8.14° was an independent risk factor for postoperative coronal decompensation.

Sodium-glucose cotransporter 2 inhibitor was initially developed and marketed as a novel oral hypoglycemic agent.Currently,it has been recommended as one of the main therapeu-tic agents for chronic heart failure by national and international guidelines,and it has a significant cardioprotective effect on heart failure with reduced ejection fraction,heart failure with im-proved ejection fraction,heart failure with mildly reduced ejec-tion fraction,and heart failure with preserved ejection fraction.However,cardiac tissues do not express sodium-glucose cotrans-porter 2,and the underlying mechanisms of clinical cardiovascu-lar benefits of sodium-glucose cotransporter 2 inhibitors remain unclear.This review intends to summarize the cardiovascular benefits of sodium-glucose cotransporter 2 inhibitors in clinical trials and the recommendations suggested by national and inter-national heart failure guidelines and to elucidate further the po-tential mechanisms of sodium-glucose cotransporter 2 inhibitor-mediated cardioprotective effects.