Objective To optimize the extraction process of Xuetong capsules and establish its quality control method. Methods The extraction process was optimized by orthogonal experiment using ethanol reflux method to investigate the effects of different factors on diphenylstilbene, aloin and extraction yield. The content of 5 anthraquinone compounds in Xuetong capsule was determined by HPLC. Results The optimal extraction process was to add 10 times ethanol, with an ethanol concentration of 70%, and extract 3 times, each time for 1 h; 5 components had a good linear relationship with peak area within a certain concentration range, r>0.999 7; The range of sample recovery rate was 93.66%-96.85%, RSD range of 1.48%-1.66%. The content determination results of the 5 components in three batches of Xuetong capsules were （0.632-0.641）, （0.660-0.681）, （1.968-1.991）, （2.547-2.580）, and （1.076-1.101） mg/g. Conclusion The method was accurate, reproducible, and highly feasible, which could be references for producing and improving the quality control standards of Xuetong capsules.

OBJECTIVE To observe the characteristics of outer membrane vesicles(OMVs)of 1 strain of hypervir-ulent Klebsiella pneumoniae(hvKp)11492.METHODS The hvKp11492,one of major clones of hvKp that were i-solated from patients with bloodstream infection in the First Medical Center of Chinese PLA General Hospital,was chosen as the research subject.The strain was identified by means of matrix-assisted laser desorption/ioniza-tion time of flight mass spectrometry and whole genome sequencing.The hvKp1 1492-OMVs were separated and purified by high speed centrifugation in combination with polymer precipitation,the morphology and particle size of the hvKp11492-OMVs were identified by transmission electron microscopy(TEM)and nanoparticle tracking a-nalysis(NTA),the proteomic characteristics were analyzed through bicinchoninic acid assay(BCA)and liquid chromatography-mass spectrometry(LC-MS)series technique,and the virulence genes were detected by PCR.RESULTS The hvKp11492-OMVs were displayed differently in size,round or oval,and complete double-layer membrane vesicles on TEM.The result of NTA showed that the average particle size of the hvKp1 1492-OMVs was 270 nm.The protein content of hvKp11492-OMVs was(2.448±0.975)μg/μl.The result of subcellular lo-calization indicated that the protein included plasmosin,intracellular membrane protein,periplasmic protein,ex-tracellular membrane protein and nuclear region protein,which were found,by the annotation of GO database,to participate in the biological processes such as oxidation-reduction,interpretation,and metabolism.The hvKp11492-OMVs contained various proteins such as GDP-L-fucose synthetase,iron ion transporter protein and ferritin that were associated with pathogenicity.In addition,the hvKp1 1492-OMVs carried with iutA,iroN,iucA and rmpA virulence genes.CONCLUSIONS The morphologic characteristics,size and proteomic characteris-tics of the hvKp11492-OMVs are identified in the study.It is concluded that the hvKp11492-OMVs carry with va-rious proteins and genes that are association with the virulence and pathogenicity.

OBJECTIVE To observe the characteristics of outer membrane vesicles(OMVs)of 1 strain of hypervir-ulent Klebsiella pneumoniae(hvKp)11492.METHODS The hvKp11492,one of major clones of hvKp that were i-solated from patients with bloodstream infection in the First Medical Center of Chinese PLA General Hospital,was chosen as the research subject.The strain was identified by means of matrix-assisted laser desorption/ioniza-tion time of flight mass spectrometry and whole genome sequencing.The hvKp1 1492-OMVs were separated and purified by high speed centrifugation in combination with polymer precipitation,the morphology and particle size of the hvKp11492-OMVs were identified by transmission electron microscopy(TEM)and nanoparticle tracking a-nalysis(NTA),the proteomic characteristics were analyzed through bicinchoninic acid assay(BCA)and liquid chromatography-mass spectrometry(LC-MS)series technique,and the virulence genes were detected by PCR.RESULTS The hvKp11492-OMVs were displayed differently in size,round or oval,and complete double-layer membrane vesicles on TEM.The result of NTA showed that the average particle size of the hvKp1 1492-OMVs was 270 nm.The protein content of hvKp11492-OMVs was(2.448±0.975)μg/μl.The result of subcellular lo-calization indicated that the protein included plasmosin,intracellular membrane protein,periplasmic protein,ex-tracellular membrane protein and nuclear region protein,which were found,by the annotation of GO database,to participate in the biological processes such as oxidation-reduction,interpretation,and metabolism.The hvKp11492-OMVs contained various proteins such as GDP-L-fucose synthetase,iron ion transporter protein and ferritin that were associated with pathogenicity.In addition,the hvKp1 1492-OMVs carried with iutA,iroN,iucA and rmpA virulence genes.CONCLUSIONS The morphologic characteristics,size and proteomic characteris-tics of the hvKp11492-OMVs are identified in the study.It is concluded that the hvKp11492-OMVs carry with va-rious proteins and genes that are association with the virulence and pathogenicity.

OBJECTIVE To evaluate the performance of commonly used molecular biological identification methods in differential identification of Citrobacter freundii complex(Cfc)and find out the differences in the mass spectra among the prevalent species of Cfc so as to facilitate the rapid and accurate identification.METHODS Totally 150 clinical strains were isolated from The First Medical Center of Chinese PLA General Hospital from 2015 to 2020 and were identified as Cfc by means of matrix assisted laser desorption ionization-time-of-flight mass spectrometry(MALDI-TOF MS),the genome data were obtained by the next generation sequencing and were compared with reference genome of Citrobacter species from National Center of Biotechnology Information(NCBI)based on aver-age nucleotide identity(ANI),phylogenetic analyses of rec N gene,concatenated alignments of four housekeeping genes(fusA,pyrG,leuS and rpoB as well as gyrB,rplB,fusA and leuS),core genome single nucleotide poly-morphism(coreSNP)and whole genome(WG),digital DNA-DNA hybridization(dDDH)analysis,ribosomal multilocus sequence typing(rMLST)and typical(strain)genome service(TYGS)analysis.ANI was set as the'gold standard' for the identification of the Cfc strains,and the capabilities of the various methods were evaluated.A certain number of strains of Citrobacter freundii(Cfr),Citrobacter braakii(Cbr)and Citrobacter portucalen-sis(Cpo)were randomly selected to obtain and compare the mass spectra so as to find out the differences in the mass spectra.RESULTS The identification capability of the Cfc strains is limited based on the available database for MALDI-TOF MS,Cpo was prone to be misidentified as Cfr or Cbr.The mass spectrum between Cpo and Cfr ranged between 2600 and 2660 m/z,the mass spectrum between Cpo and Cbr ranged between 5200 and 5300 m/z,showing significant difference.The concatenated alignments of four housekeeping genes gyrB,rplB,fusA and leuS,CoreSNP and WG phylogenetic analysis had high accurate rate of identification.CONCLUSIONS The meth-ods have certain limitations in identification of Cfc.The joint application of ANI with the methods such as WG phylogenetic analysis,update of mass spectrum database and timely establishment of database can achieve the ac-curate identification of the species of Cfc.

OBJECTIVE To explore the distribution of pathogens isolated from the thoracoscopic lobectomy patients with postoperative pulmonary infections and observe the expressions of maternal expression gene 3(MEG3)/microribonucleic acid-223(miR-223)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)axis in peripheral blood.METHODS Totally 48 lung cancer patients who underwent thoracoscopic lobec-tomy and had postoperative pulmonary infections in the First Affiliated Hospital of Guangzhou University of Chi-nese Medicine from Jun.2021 to Jun.2024 were assigned as the infection group,meanwhile,31 lung cancer pa-tients who underwent thoracoscopic lobectomy but did not have postoperative infections were chosen as the no in-fection group.The sputum specimens were collected from the infection group,the distribution of isolated patho-gens was analyzed.The clinical data and the expression MEG3,miR-223 and NLRP3 were observed and compared between the two groups.The values of MEG3,miR-223 and NLRP3 in prediction of the postoperative infections were analyzed by receiver operating characteristic(ROC)curves.RESULTS Totally 63 strains of pathogens were i-solated from the 48 thoracoscopic lobectomy patients with postoperative pulmonary infections,33(52.38％)of which were gram-negative bacteria,25(39.68％)were gram-positive bacteria,and 5(7.94％)were fungi.There were significant differences in age,diabetes mellitus,chronic obstructive pulmonary disease(COPD),operation duration,and expression levels of MEG3,miR-223 and NLRP3 between the infection group and the no infection group(P＜0.05).ROC curve analysis showed that the areas under the curves of MEG3,miR-223 and NLRP3 and the joint detection of the three indexes were 0.861,0.760,0.912 and 0.968,respectively,in prediction of the post-operative pulmonary infections,and the predictive efficiency of the joint detection was highest(P＜0.05).CONCLUSIONS The gram-negative bacteria are dominant among the pathogens isolated from the thoracoscopic lo-bectomy patients with postoperative pulmonary infections,the occurrence and progression of the infections may be closely associated with the activation of MEG3/miR-223/NLRP3 axis.The joint detection of the three indexes can effectively predict the postoperative pulmonary infections in the thoracoscopic lobectomy patients.

OBJECTIVE To evaluate the performance of commonly used molecular biological identification methods in differential identification of Citrobacter freundii complex(Cfc)and find out the differences in the mass spectra among the prevalent species of Cfc so as to facilitate the rapid and accurate identification.METHODS Totally 150 clinical strains were isolated from The First Medical Center of Chinese PLA General Hospital from 2015 to 2020 and were identified as Cfc by means of matrix assisted laser desorption ionization-time-of-flight mass spectrometry(MALDI-TOF MS),the genome data were obtained by the next generation sequencing and were compared with reference genome of Citrobacter species from National Center of Biotechnology Information(NCBI)based on aver-age nucleotide identity(ANI),phylogenetic analyses of rec N gene,concatenated alignments of four housekeeping genes(fusA,pyrG,leuS and rpoB as well as gyrB,rplB,fusA and leuS),core genome single nucleotide poly-morphism(coreSNP)and whole genome(WG),digital DNA-DNA hybridization(dDDH)analysis,ribosomal multilocus sequence typing(rMLST)and typical(strain)genome service(TYGS)analysis.ANI was set as the'gold standard' for the identification of the Cfc strains,and the capabilities of the various methods were evaluated.A certain number of strains of Citrobacter freundii(Cfr),Citrobacter braakii(Cbr)and Citrobacter portucalen-sis(Cpo)were randomly selected to obtain and compare the mass spectra so as to find out the differences in the mass spectra.RESULTS The identification capability of the Cfc strains is limited based on the available database for MALDI-TOF MS,Cpo was prone to be misidentified as Cfr or Cbr.The mass spectrum between Cpo and Cfr ranged between 2600 and 2660 m/z,the mass spectrum between Cpo and Cbr ranged between 5200 and 5300 m/z,showing significant difference.The concatenated alignments of four housekeeping genes gyrB,rplB,fusA and leuS,CoreSNP and WG phylogenetic analysis had high accurate rate of identification.CONCLUSIONS The meth-ods have certain limitations in identification of Cfc.The joint application of ANI with the methods such as WG phylogenetic analysis,update of mass spectrum database and timely establishment of database can achieve the ac-curate identification of the species of Cfc.

OBJECTIVE To explore the distribution of pathogens isolated from the thoracoscopic lobectomy patients with postoperative pulmonary infections and observe the expressions of maternal expression gene 3(MEG3)/microribonucleic acid-223(miR-223)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)axis in peripheral blood.METHODS Totally 48 lung cancer patients who underwent thoracoscopic lobec-tomy and had postoperative pulmonary infections in the First Affiliated Hospital of Guangzhou University of Chi-nese Medicine from Jun.2021 to Jun.2024 were assigned as the infection group,meanwhile,31 lung cancer pa-tients who underwent thoracoscopic lobectomy but did not have postoperative infections were chosen as the no in-fection group.The sputum specimens were collected from the infection group,the distribution of isolated patho-gens was analyzed.The clinical data and the expression MEG3,miR-223 and NLRP3 were observed and compared between the two groups.The values of MEG3,miR-223 and NLRP3 in prediction of the postoperative infections were analyzed by receiver operating characteristic(ROC)curves.RESULTS Totally 63 strains of pathogens were i-solated from the 48 thoracoscopic lobectomy patients with postoperative pulmonary infections,33(52.38％)of which were gram-negative bacteria,25(39.68％)were gram-positive bacteria,and 5(7.94％)were fungi.There were significant differences in age,diabetes mellitus,chronic obstructive pulmonary disease(COPD),operation duration,and expression levels of MEG3,miR-223 and NLRP3 between the infection group and the no infection group(P＜0.05).ROC curve analysis showed that the areas under the curves of MEG3,miR-223 and NLRP3 and the joint detection of the three indexes were 0.861,0.760,0.912 and 0.968,respectively,in prediction of the post-operative pulmonary infections,and the predictive efficiency of the joint detection was highest(P＜0.05).CONCLUSIONS The gram-negative bacteria are dominant among the pathogens isolated from the thoracoscopic lo-bectomy patients with postoperative pulmonary infections,the occurrence and progression of the infections may be closely associated with the activation of MEG3/miR-223/NLRP3 axis.The joint detection of the three indexes can effectively predict the postoperative pulmonary infections in the thoracoscopic lobectomy patients.

Objective To establish the method of simultaneous determination of four main components of Danggui Liuhuang Decoction,including phellodendrine,palmatine,calycosin,and ferulic acid and provide reference for the quality control of Danggui Liuhuang Decoction.Methods Based on the HPLC-MS/MS analysis method,the positive ion data acquisition mode were adopted for the mass spectrometry detection and the four main components were quantified with multiple reaction monitoring mode(MRM)by ESI source.The chromatographic column was Agilent Extend-C18(5 μm,4.6 mm×250 mm),and gradient elution was performed with methanol and 0.5%formic acid in water.Results The linear range of phellodendrine was from 2-200 nmol/ml,and the linear range of palmatine,calycosin and ferulic acid was from 20-2 000 nmol/ml.The contents of the four components in the seven batches of Danggui Liuhuang Decoction were relatively stable,among which ferulic acid was mainly found in Phellodendrine and Coptidis;Phellodendrine was only detected in cortex phellodendri;the content of calycosin in Scutellaria baicalensis and Astragalus was higher;palmatine was detected in both Phellodendron and Astragalus.Conclusion The method had high sensitivity,good specificity and sample stability,which could meet the requirements of quantitative analysis of Traditional Chinese Medicine compounds,and could provide reference for further pharmacokinetics study on the content changes of traditional Chinese medicine compounds in biological samples.

Purpose: The objective of this study was to propose a deep-learning model for the detection of the mandibular canal on dental panoramic radiographs. Materials and Methods: A total of 2,100 panoramic radiographs (PANs) were collected from 3 different machines: RAYSCAN Alpha (n=700, PAN A), OP-100 (n=700, PAN B), and CS8100 (n=700, PAN C). Initially, an oral and maxillofacial radiologist coarsely annotated the mandibular canals. For deep learning analysis, convolutional neural networks (CNNs) utilizing U-Net architecture were employed for automated canal segmentation. Seven independent networks were trained using training sets representing all possible combinations of the 3 groups. These networks were then assessed using a hold-out test dataset. Results: Among the 7 networks evaluated, the network trained with all 3 available groups achieved an average precision of 90.6%, a recall of 87.4%, and a Dice similarity coefficient (DSC) of 88.9%. The 3 networks trained using each of the 3 possible 2-group combinations also demonstrated reliable performance for mandibular canal segmentation, as follows: 1) PAN A and B exhibited a mean DSC of 87.9%, 2) PAN A and C displayed a mean DSC of 87.8%, and 3) PAN B and C demonstrated a mean DSC of 88.4%. Conclusion This multi-device study indicated that the examined CNN-based deep learning approach can achieve excellent canal segmentation performance, with a DSC exceeding 88%. Furthermore, the study highlighted the importance of considering the characteristics of panoramic radiographs when developing a robust deep-learning network, rather than depending solely on the size of the dataset.

In December 2020, a 39-year female was admitted in Guangzhou Heping Orthopaedic Hospital with severely and destructively mangled both forearms. There was a segmental destruction of left wrist and an oblique destruction of right palm together with the right thumb and index finger. In the emergency surgery, the left palm was split at the first web. Then the left thumb was transferred to the first metacarpal bone of right hand for reconstruction of right thumb. The 2nd to 4th metacarpal bones of the left palm were transferred to the distal carpal bones of right wrist for reconstruction of right hand. The superficial veins of forearm were freed and transferred to bridge the defects of arteries and veins in the right hand. A right anteriolateral thigh perforator flap (ALTPF) was taken to reconstruct the soft tissue defects of right hand. The ring and little fingers of right hand were amputated and transferred to the left distal ulna and distal radius of the stump of remaining left forearm, thereby a chimeric left "forearm-hand" was reconstructed. After 20 months of follow-up, the both reconstructed hands and the flaps survived well without obvious bloat, and the appearance of the digits was full. The two-point discrimination (TPD) of both reconstructed hands was restored to 10-12 mm, and a partial pinching and griping functions were restored. The reconstructed hands were able to fulfil the essential requirements of daily life. Only linear scars were left in the donor sites of right forearm and right thigh.