[Objective] To report a new technique that combines microfracture surgery under local anesthesia with injection of platelet-rich plasma (PRP) at the fracture site, so as to improve fracture healing rates. [Methods] Data from patients who visited our hospital from March 2020 to June 2023 and underwent the treatment for delayed union of limb fractures were retrospectively analyzed. Under local infiltrative anesthesia, with the assistance of a C-arm X-ray machine or ultrasound, percutaneous loosening was done at the fracture site and the medullary cavity, followed by cortical drilling around the fracture. The previously prepared PRP was then injected locally at the fracture site. Patients were followed up and their postoperative recovery was recorded. [Results] All patients were followed up, and the fracture healing rate was 94.12% (16/17), with an average healing duration of (5.88±2.50) months. None of the patients experienced any neural or vascular injuries, nor adverse events such as wound infections or osteomyelitis. Before the operation and at the last follow-up, the patients' pain visual analogue scores were (5.12±1.11) vs (0.71±1.21) respectively. The postoperative VAS scores showed a significant decrease compared to preoperative values (P<0.05). The excellent and good rate for limb function on the affected side was 88.24% (14/17) at the last follow-up, which was a significant increase from 0.00% before surgery (P<0.05). [Conclusion] The injection of PRP at the fracture site combined with microfracture surgery at the fracture site is minimally invasive, simple to perform, and well-accepted by patients. It has demonstrated some clinical efficacy in treating delayed fracture healing.

Objective To perform single-molecule, real-time sequencing of ceftazidime-avibactam (CAZ-AVI)-resistant Pseudomonas aeruginosa and to investigate the mechanism underlying ceftazidime-avibactam resistance in P. aeruginosa. Methods The susceptibility of 89 P. aeruginosa isolates randomly sampled from clinical specimens in Sanming First Hospital Affiliated to Fujian Medical University from November 2021 through July 2023 to common antimicrobial agents was tested, and the minimum inhibitory concentration (MIC) of CAZ-AVI was determined against P. aeruginosa with a broth microdilution assay, with CAZ-AVI MICs of 8 mg/L and lower defined as susceptible and 16 mg/L and higher as resistant. The expression of drug-resistant genes ampC, oxa-488, oprD, mexA, oxa-10, oxa-14, vim and tem was quantified in P. aeruginosa using a real-time quantitative reverse transcription PCR (qPCR) assay. CAZ-AVI-susceptible and -resistant P. aeruginosa isolates from the same case were selected for PacBio single-molecule, real-time sequencing, and sequencing results were subjected to genome structure and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotations. Results The 89 P. aeruginosa isolates showed a relatively high level of resistance to meropenem (75.28%) and imipenem (74.16%) and the highest susceptibility to amikacin (91.01%). There were 49 CAZ-AVI-resistant P. aeruginosa isolates and 40 susceptible isolates. qPCR assay detected lower oprD gene expression in CAZ-AVI-resistant P. aeruginosa isolates [0.104 (2.385)] than in susceptible isolates [0.551 (17.885)] (Z = -2.958, P < 0.01), and there were no significant differences between CAZ-AVI-susceptible and -resistant P. aeruginosa isolates in terms of ampC, oxa-488, mexA or tem gene expression (all P values > 0.05), while oxa-10, oxa-14 and vim gene was expressed in few P. aeruginosa isolates. There were 1 729, 3 936, 3 737 and 3 955 genes in CAZ-AVI-resistant P. aeruginosa isolates PA-762 and PA-M174 and susceptible isolates PA-885 and PA-808 that were annotated to GO terms, with the highest numbers of genes enriched in the molecular function of catalytic activity, high numbers of genes enriched in biological processes of metabolic process, single-organism process and cellular process, and high numbers of genes enriched in cellular components of cell and cell membranes. There were 1 803, 4 084, 3 915 and 4 066 genes in the PA-762, PA-M174, PA-885 and PA-808 isolates enriched in the KEGG signaling pathway, and the majority of genes were enriched in four primary signaling pathways of metabolism, genetic information processing, environmental information processing and cellular process, with the highest number of genes associated with metabolic pathways. Both CAZ-AVI-resistant P. aeruginosa isolates PA-762 and PA-M174 carried multiple efflux pumps systems, including MexAB-OprM, MexCD-OprJ, MexEF-OprN and MexXY-OprM. Single nucleotide substitution was found at position 169 in the DNA sequence of the PA-762 isolate, leading to substitution of serine for glycine at position 57 in the protein sequence, and there are deletions of two bases at positions 307 and 308 in the DNA sequence of the PA-M174 isolate, leading to substitution of threonine for arginine at position 103 in the protein sequence. Conclusion Mutation or downregulation of oprD gene may lead to CAZ-AVI resistance in P. aeruginosa.

Objective:To investigate whether biochanin A(BCA)has a protective effect against dextran sodium sulfate(DSS)-in-duced ulcerative colitis(UC)in mice.Methods:Thirty C57BL/6N mice were randomly divided into normal control group,DSS model group,sulfasalazine(SASP)-positive drug control group,and low/medium/high-dose(5 mg/kg,10 mg/kg,and 20 mg/kg)BCA groups.The mouse model of UC was induced by administering 2.5%DSS aqueous solution for 7 days.During the experimental period,both the normal control and model groups were given 0.5%carboxymethyl cellulose sodium solution daily by gavage.The positive control group was given 100 mg/kg SASP,while the BCA groups were given BCA suspensions at doses of 5 mg/kg,10 mg/kg,and 20 mg/kg.The ad-ministration lasted for 10 days.Body weight changes and fecal status of the mice were recorded every day;the colon was dissected,col-lected,and measured for its length.The colon was stained with Hematoxylin-Eosin for pathomorphological study.Quantitative poly-merase chain reaction was used to determine the levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-10(IL-10)in the colon.Immunofluorescence was used to determine the expression of tight junction proteins,zonula occluden-1(ZO-1)and occludin,in the colon.Results:It showed that 5 mg/kg and 10 mg/kg BCA significantly alleviated weight loss in mice with UC,while 5 mg/kg,10 mg/kg,and 20 mg/kg BCA reduced the disease activity index scores.Additionally,BCA showed similar effects to SASP in improving the structure and reducing the shortening of the colon in mice with UC.Compared with the model group,all BCA groups had significantly decreased TNF-α(P=0.024、P=0.060、P=0.003)and IL-6(P=0.002、P<0.001、P<0.001)and significantly increased IL-10(P=0.006、P=0.003、P<0.001),with varying degrees of up-regulated expression of tight junction proteins.Conclusion:BCA can effectively alleviate DSS-induced symptoms,reduce intes-tinal damage,and protect the intestinal barrier in mice with UC.

Objective:This study aimed to compare short-chain fatty acid (SCFA) levels in saliva between patients with laryngopharyngeal reflux disease (LPRD) and healthy controls, and to explore the relationship between these SCFAs and the salivary microbiota.Methods:A retrospective case-control study was conducted, enrolling 36 patients with laryngopharyngeal reflux disease (LPRD) who visited the Department of Otorhinolaryngology Head and Neck Surgery, the Eighth Medical Center, Chinese PLA General Hospital between February and November 2023. All patients were diagnosed via pharyngeal pH monitoring. The LPRD group included 30 males and 6 females, aged 20-53 years (30.61±7.83 years). In addition, 39 healthy volunteers were recruited as the control group, comprising 25 males and 14 females, aged 18–58 years (28.64±7.97 years). Unstimulated mixed saliva samples were collected from all participants. Concentrations of eight SCFAs (acetic acid, propionic acid, isobutyric acid, butyric acid, valeric acid, isovaleric acid, hexanoic acid, and heptanoic acid) in saliva were quantified using gas chromatography-mass spectrometry (GC-MS). Salivary DNA was extracted, followed by amplification and sequencing of the 16S rRNA gene to analyze the microbiota composition at the genus level. The SCFA concentrations and the differences in bacterial species between the LPRD and control groups were compared, and the correlation between SCFA concentrations and the relative abundance of different bacterial genera in the salivary microbiota was analyzed. All statistical analyses were performed using R version 3.6.1 and SPSS version 26.0, while, microbiome analyses were conducted using R language.Results:Salivary hexanoic acid concentration was significantly higher in the LPRD group than in the control group [(29.50±19.61) ng/ml vs. (10.15±3.65) ng/ml; t=-2.72, P<0.05]. Significant differences in the relative abundance of 17 bacterial genera were observed between the two groups ( P<0.05), including Prevotella, Butyrivibrio, Streptococcus, and Actinomyces. Correlation analysis revealed that hexanoic acid concentration was significantly positively correlated with the abundance of Butyrivibrio (γ=0.73, P<0.05) and Streptococcus (γ=0.78, P<0.05), while showing a significant negative correlation with Actinomyces (γ=-0.73, P<0.05). Conclusion:Elevated salivary hexanoic acid levels may be associated with the development of LPRD. Dysbiosis of the salivary microbiota might contribute to LPRD pathogenesis by altering the concentrations of SCFA, particularly hexanoic acid.

Objective To explore the value of amide proton transfer(APT)imaging combined with diffusion weighted imaging(DWI)in the assessment of neurovascular invasion(NVI)of stage T3/T4 rectal cancer.Methods The clinical and MR imaging data of 46 patients with rectal cancer were analyzed retrospectively and divided into NVI positive group and NVI negative group according to the pathological results of NVI.The differences of APT values[maximum APT(APTmax)value,minimum APT(APTmin)value,mean APT(APTmean)value]and apparent diffusion coefficient(ADC)value between the two groups were compared,and the diagnostic efficiency was analyzed by receiver operating characteristic(ROC)curve.Results The APTmax and APTmean values in the NVI positive group were significantly higher than those in the NVI negative group,while the ADC value in the NVI positive group was significantly lower than that in the NVI negative group(P＜0.05).The area under the curve(AUC)of APTmax,APTmean and ADC values for NVI identification were 0.717,0.722 and 0.751,respectively.The AUC of the three indexes combined to identify NVI was 0.858.The combined AUC of the three indexes was larger than that of APTmax,APTmean and ADC alone(P＜0.05).Conclusion APT imaging combined with DWI has a high value in the evaluation of NVI of stage T3/T4 rectal cancer,which can provide guidance for clinical treatment.

Objective To explore the value of amide proton transfer(APT)imaging combined with diffusion weighted imaging(DWI)in the assessment of neurovascular invasion(NVI)of stage T3/T4 rectal cancer.Methods The clinical and MR imaging data of 46 patients with rectal cancer were analyzed retrospectively and divided into NVI positive group and NVI negative group according to the pathological results of NVI.The differences of APT values[maximum APT(APTmax)value,minimum APT(APTmin)value,mean APT(APTmean)value]and apparent diffusion coefficient(ADC)value between the two groups were compared,and the diagnostic efficiency was analyzed by receiver operating characteristic(ROC)curve.Results The APTmax and APTmean values in the NVI positive group were significantly higher than those in the NVI negative group,while the ADC value in the NVI positive group was significantly lower than that in the NVI negative group(P＜0.05).The area under the curve(AUC)of APTmax,APTmean and ADC values for NVI identification were 0.717,0.722 and 0.751,respectively.The AUC of the three indexes combined to identify NVI was 0.858.The combined AUC of the three indexes was larger than that of APTmax,APTmean and ADC alone(P＜0.05).Conclusion APT imaging combined with DWI has a high value in the evaluation of NVI of stage T3/T4 rectal cancer,which can provide guidance for clinical treatment.

Objective:This study aimed to compare short-chain fatty acid (SCFA) levels in saliva between patients with laryngopharyngeal reflux disease (LPRD) and healthy controls, and to explore the relationship between these SCFAs and the salivary microbiota.Methods:A retrospective case-control study was conducted, enrolling 36 patients with laryngopharyngeal reflux disease (LPRD) who visited the Department of Otorhinolaryngology Head and Neck Surgery, the Eighth Medical Center, Chinese PLA General Hospital between February and November 2023. All patients were diagnosed via pharyngeal pH monitoring. The LPRD group included 30 males and 6 females, aged 20-53 years (30.61±7.83 years). In addition, 39 healthy volunteers were recruited as the control group, comprising 25 males and 14 females, aged 18–58 years (28.64±7.97 years). Unstimulated mixed saliva samples were collected from all participants. Concentrations of eight SCFAs (acetic acid, propionic acid, isobutyric acid, butyric acid, valeric acid, isovaleric acid, hexanoic acid, and heptanoic acid) in saliva were quantified using gas chromatography-mass spectrometry (GC-MS). Salivary DNA was extracted, followed by amplification and sequencing of the 16S rRNA gene to analyze the microbiota composition at the genus level. The SCFA concentrations and the differences in bacterial species between the LPRD and control groups were compared, and the correlation between SCFA concentrations and the relative abundance of different bacterial genera in the salivary microbiota was analyzed. All statistical analyses were performed using R version 3.6.1 and SPSS version 26.0, while, microbiome analyses were conducted using R language.Results:Salivary hexanoic acid concentration was significantly higher in the LPRD group than in the control group [(29.50±19.61) ng/ml vs. (10.15±3.65) ng/ml; t=-2.72, P<0.05]. Significant differences in the relative abundance of 17 bacterial genera were observed between the two groups ( P<0.05), including Prevotella, Butyrivibrio, Streptococcus, and Actinomyces. Correlation analysis revealed that hexanoic acid concentration was significantly positively correlated with the abundance of Butyrivibrio (γ=0.73, P<0.05) and Streptococcus (γ=0.78, P<0.05), while showing a significant negative correlation with Actinomyces (γ=-0.73, P<0.05). Conclusion:Elevated salivary hexanoic acid levels may be associated with the development of LPRD. Dysbiosis of the salivary microbiota might contribute to LPRD pathogenesis by altering the concentrations of SCFA, particularly hexanoic acid.

Objective To discuss the value of clinical radiomic nomogram(CRN)and deep convolutional neural network(DCNN)in distinguishing atypical pulmonary hamartoma(APH)from atypical lung adenocarcinoma(ALA).Methods A total of 307 patients were retrospectively recruited from two institutions.Patients in institu-tion 1 were randomly divided into the training(n=184:APH=97,ALA=87)and internal validation sets(n=79:APH=41,ALA=38)in a ratio of 7∶3,and patients in institution 2 were assigned as the external validation set(n=44:APH=23,ALA=21).A CRN model and a DCNN model were established,respectively,and the performances of two models were compared by delong test and receiver operating characteristic(ROC)curves.A human-machine competition was conducted to evaluate the value of AI in the Lung-RADS classification.Results The areas under the curve(AUCs)of DCNN model were higher than those of CRN model in the training,internal and external validation sets(0.983 vs 0.968,0.973 vs 0.953,and 0.942 vs 0.932,respectively),however,the differences were not statistically significant(p=0.23,0.31 and 0.34,respectively).With a radiologist-AI com-petition experiment,AI tended to downgrade more Lung-RADS categories in APH and affirm more Lung-RADS cat-egories in ALA than radiologists.Conclusion Both DCNN and CRN have higher value in distinguishing APH from ALA,with the former performing better.AI is superior to radiologists in evaluating the Lung-RADS classification of pulmonary nodules.

Objective To investigate the mechanism by which swertiamarin(STM)ameliorates CD-like colitis in mice.Methods A Caco-2 cell model of TNF-α-stimulated apoptosis was established and divided into three groups:Con,TNF-α and STM,and the effects of STM on apoptosis and barrier function were assessed by Tunel staining,western blotting,immunofluorescence,and transepithelial electric resistance(TEER).A mouse model of 2,4,6-trinitrobenzenesulfonic acid(TNBS)-induced CD-like colitis was established to assess the effects of STM on colitis,intestinal barrier function and epithelial cell apoptosis.The regulatory role of the PI3K/AKT pathway in STM-induced resistance to intestinal epithelial cell apoptosis was investigated in both the cell model and mouse models.Results TUNEL staining showed that in Caco-2 cells with TNF-α stimulation,STM treatment significantly reduced the percentage of TUNEL-stained cells(P<0.05).STM obviously reduced TNF-α-induced enhancement of cleaved-caspase 3 and Bax expressions(P<0.05),increased Bcl-2 expression(P<0.05),protected intestinal barrier integrity and function by restoring transepithelial electrical resistance(TEER)of the cells,promoted normal localization and expressions of the tight junction proteins(ZO1 and claudin 1)(P<0.05),and inhibited the expression of pro-inflammatory factors(IL-6 and CCL3)(P<0.05)in TNF-α-stimulated Caco-2 cells.In the mouse models,STM significantly alleviated TNBS-induced CD-like colitis and intestinal barrier dysfunction(P<0.05)as shown by improved weight loss,lowered Disease Activity Index(DAI)score and inflammation score,reduction of IL-6 and CCL3 release,and restoration of intestinal barrier permeability,colonic TEER,bacterial translocation,and localization and expressions of the tight junction proteins.Mechanistically,STM inhibited the expressions of p-PI3K and p-AKT in both the cell model and mouse model(P<0.05),and treatment with 740Y-P(a PI3K/AKT pathway activator)significantly attenuated the inhibitory effect of STM on TNF-α-induced apoptosis in Caco-2 cells(P<0.05).Conclusion STM inhibits intestinal epithelial cell apoptosis at least in part by suppressing activation of the PI3K/AKT pathway to ameliorate intestinal barrier dysfunction and colitis in mice.

Objective To investigate the mechanism by which swertiamarin(STM)ameliorates CD-like colitis in mice.Methods A Caco-2 cell model of TNF-α-stimulated apoptosis was established and divided into three groups:Con,TNF-α and STM,and the effects of STM on apoptosis and barrier function were assessed by Tunel staining,western blotting,immunofluorescence,and transepithelial electric resistance(TEER).A mouse model of 2,4,6-trinitrobenzenesulfonic acid(TNBS)-induced CD-like colitis was established to assess the effects of STM on colitis,intestinal barrier function and epithelial cell apoptosis.The regulatory role of the PI3K/AKT pathway in STM-induced resistance to intestinal epithelial cell apoptosis was investigated in both the cell model and mouse models.Results TUNEL staining showed that in Caco-2 cells with TNF-α stimulation,STM treatment significantly reduced the percentage of TUNEL-stained cells(P<0.05).STM obviously reduced TNF-α-induced enhancement of cleaved-caspase 3 and Bax expressions(P<0.05),increased Bcl-2 expression(P<0.05),protected intestinal barrier integrity and function by restoring transepithelial electrical resistance(TEER)of the cells,promoted normal localization and expressions of the tight junction proteins(ZO1 and claudin 1)(P<0.05),and inhibited the expression of pro-inflammatory factors(IL-6 and CCL3)(P<0.05)in TNF-α-stimulated Caco-2 cells.In the mouse models,STM significantly alleviated TNBS-induced CD-like colitis and intestinal barrier dysfunction(P<0.05)as shown by improved weight loss,lowered Disease Activity Index(DAI)score and inflammation score,reduction of IL-6 and CCL3 release,and restoration of intestinal barrier permeability,colonic TEER,bacterial translocation,and localization and expressions of the tight junction proteins.Mechanistically,STM inhibited the expressions of p-PI3K and p-AKT in both the cell model and mouse model(P<0.05),and treatment with 740Y-P(a PI3K/AKT pathway activator)significantly attenuated the inhibitory effect of STM on TNF-α-induced apoptosis in Caco-2 cells(P<0.05).Conclusion STM inhibits intestinal epithelial cell apoptosis at least in part by suppressing activation of the PI3K/AKT pathway to ameliorate intestinal barrier dysfunction and colitis in mice.