BACKGROUND: Endoscopic sphincterotomy (EST) is widely used to treat common bile duct stones (CBDS); however, long-term studies have revealed the increasing incidence of recurrent CBDS after EST. Loss of sphincter of Oddi function after EST was the main cause of recurrent CBDS. Reparation of the sphincter of Oddi is therefore crucial. This study aims to investigate the effectiveness and safety of endoclip papilloplasty (ECPP) for repairing the sphincter of Oddi and elucidate its mechanism. METHODS: Eight healthy Bama minipigs were randomly divided into the EST group and the ECPP group at a 1:1 ratio, and bile samples were collected before endoscopy and 6 months later. All minipigs underwent transabdominal biliary ultrasonography for the diagnosis of cholelithiasis 6 months after endoscopy. The biliary microbiota composition and alpha and beta diversity were analyzed by 16S ribosomal RNA gene sequencing. Differential metabolites were analyzed by bile acid metabolomics to explore the predictive indicators of cholelithiasis. RESULTS: Three minipigs were diagnosed with cholelithiasis in the EST group, while none in the ECPP group showed cholelithiasis. The biliary Firmicutes/Bacteroidota (F/B) ratio was increased after EST and decreased after ECPP. The Chao1 and observed species index significantly decreased 6 months after EST ( P  = 0.017 and 0.018, respectively); however, the biliary α-diversity was similar before and 6 months after ECPP. The β-diversity significantly differed in the EST group before and 6 months after EST, as well as in the ECPP group before and 6 months after ECPP (analysis of similarities [ANOSIM]: R  = 0.917, P  = 0.040; R  = 0.740, P  = 0.035; respectively). Glycolithocholic acid (GLCA) and taurolithocholic acid (TLCA) accumulated in bile 6 months after EST. CONCLUSIONS ECPP has less impact on the biliary microenvironment than EST and prevents duodenobiliary reflux by repairing the sphincter of Oddi. The bile levels of GLCA and TLCA may be used to predict the risk of cholelithiasis.
Animals ; Swine, Miniature ; Swine ; Cholelithiasis/prevention & control* ; Sphincterotomy, Endoscopic/methods* ; Sphincter of Oddi/surgery* ; Female ; Male

Objective:To prepare seven standard strains of 2019 novel coronavirus (2019-nCoV), including wild type (WT) strain, Beta variant, Delta variant, Omicron variants (BA.2, BA.5, BQ.1, XBB.1 branches), which could be used to apply for national standard strains.Methods:According to cytopathic effect (CPE), virus titer, whole-virus-genome-sequencing and detection of mycoplasma, the basic biological characteristics of clonal isolation were determined through plaque purification technology.Results:The CPE was mainly characterized by cell shrinkage and exfoliation in Vero cells after infection with seven 2019-nCoV clonal isolations (WT, Beta, Delta, Omicron, BA.2, BA.5, BQ.1 and XBB.1 branches). The result of mycoplasma detection were negative and titers of the clonal isolation from 2nd to 5th generations were stable at 10 5-10 8 TCID 50/ml; the electron microscope showed that the virions were all round or elliptical, with a diameter between 60 nm and 140 nm. The subtypes of 7 strains were identified by whole-virus-genome-sequencing and phylogenetic analysis, with genomic stability after the fifth successive generations of clonal isolation. Conclusions:The series clonal isolation of 2019-nCoV with typical CPE of coronavirus, clear morphological structure, good viral activity and stable genetic characteristics were prepared, and they could be used to apply for national standard strains.

Objective:To recheck the Histoplasma capsulatum and Coccidioides sp. strains of the past by molecular identification and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS). Methods:The phylogenetic relationships among the ten Histoplasma capsulatum isolates and reference strains were analyzed by multilocus sequence typing (MLST). Based on the Coi region, Coccidioides posadasii was distinguished from Coccidioides immitis accurately. MALDI-TOF MS was used to set up the MALDI-TOF MS database of Histoplasma capsulatum and Coccidioides sp. for rapid identification. In addition, hierarchical clustering of spectra was compared with MLST. Results:An unrooted dendrogram constructed with MLST showed that ten individuals of Histoplasma capsulatum were divided into three clades: Eurasia clade, Australia clade and North American class 2 clade, in agreement with the establishment by MALDI-TOF MS cluster analysis. All individuals of Coccidioides sp. were identified as Coccidioides posadasii with Coi region primers. The in-house MALDI-TOF MS database of Histoplasma capsulatum and Coccidioides posadasii was expanded and reached an identified accuracy of 100%. Conclusions:We improve the recognition of Histoplasma capsulatum and Coccidioides posadasii by molecular pathways which shows the major species or clades in Chinese mainland. The in-house MALDI-TOF MS database of Histoplasma capsulatum and Coccidioides posadasii can provide a new efficient way to identify those pathogens rapidly.

Objective:To understand the clinical prevalence and drug resistance of Raoultella ornithinolytica, and to analyze the drug-resistant characteristics of carbapenem-resistant strains. Methods:A total of 83 clinical isolates of R.ornithinolytica, identified using matrix-assisted laser desorption ionization time-of-flight mass spectrometry between May 2015 and December 2020, were collected. Clinical information, drug-resistant phenotype, and carbapenemase gene of the carbapenem resistant strains were analyzed. Homology of the resistant strains was examined by pulse-field gel electrophoresis (PFGE). The molecular characteristics of plasmids, drug resistance genes and virulence genes were determined by whole genome sequencing (WGS), and a phylogenetic tree was constructed based on core single-nucleotide polymorphisms(core-SNPs) for evolutionary analysis of the carbapenem-resistant strains. Results:All 83 clinical isolates were confirmed as R.ornithinolytica, of which 11 strains were resistant to carbapenem antibiotics. The resistant strains were mainly sourced from the drainage fluid and wound secretions of patients in surgery, intensive care unit and oncology department. In addition to carbapenems resistances, the 11 strains were also resistant to cephalosporins, cephamycins, and aztreonam, while maintaining good in vitro activity to levofloxacin, tigecycline and colistin. Carbapenemase gene detection showed that 8 strains carried the bla IMP-4, 3 carried bla KPC-2, of which 1 strain carried both bla IMP-4 and bla KPC-2, and 2 strains carried both bla IMP-4 and bla NDM-1. PFGE results showed 7 banding types and 4 epidemic strains. WGS identified plasmid replicon types carried by the 11 strains of R. ornithinolyticus including IncFIA, IncFIB, IncHI1B, IncU, repB and Col, along with 10 additional types of drug resistance genes. Iron carrier related virulence genes entB, fyu, ybt and irp were detected in all carbapenem-resistant strains. Phylogenetic analysis showed that Ro6348 and Ro6050, Ro7401 and Ro7242, Ro8647 and Ro5832, and Ro9907 and Ro8856 were grouped together, while Ro8454, Ro7349 and Ro7326 were located on different branches. Conclusions:The overall drug resistance of R. ornithinolytica closely resembles Klebsiella pneumoniae and carries a variety of drug-resistant genes, but differs from domestic K.pneumoniae in that its carbapenemase is mainly IMP-4. PFGE and phylogenetic analysis results suggest that the strain has a tendency of mutual transmission in hospital, which deserves more attention.

Objective:To investigate the variation of reference ranges of hemodynamic parameters in normal pregnancy and their relation to maternal basic characteristics.Methods:A total of 598 healthy pregnant women who underwent regular prenatal examination at the Third Affiliated Hospital of Guangzhou Medical University from January to December 2023 were prospectively enrolled, and noninvasive hemodynamic monitors were used to detect changes in hemodynamic parameters of the pregnant women with the week of gestation, including cardiac output (CO), stroke volume (SV), thoracic fluid content (TFC), systemic vascular resistance (SVR), mean arterial pressure (MAP), and heart rate (HR). Relationships between hemodynamic parameters and maternal basic characteristics, including age, height, and weight, were analyzed using restricted cubic spline.Results:(1) CO ( r=0.155, P<0.001), TFC ( r=0.338, P<0.001), MAP ( r=0.204, P<0.001), and HR ( r=0.352, P<0.001) were positively correlated with the week of gestation, and SV was negatively correlated with the week of gestation ( r=-0.158, P<0.001). There was no significant correlation between SVR and gestational age ( r=-0.051, P=0.258). (2) CO exhibited a positive correlation with maternal height and weight (all P<0.001). The taller and heavier of pregnant women, the higher their CO. A linear relationship was observed between maternal weight and SV, MAP and HR (all P<0.01). As maternal weight increased, SV, MAP and HR showed an upward trend. Furthermore, there was an inverse association between maternal age and SVR ( P<0.001). (3) There was a significant nonlinear association observed between TFC and body mass index during pregnancy ( P<0.05). Additionally, a nonlinear relationship was found between SVR and MAP in relation to maternal age (all P<0.05). Notably, when the age exceeded 31 years old, there was an evident upward trend observed in both SVR and MAP. Conclusions:The hemodynamic parameters of normal pregnant women are influenced by their height, body weight, and age. It is advisable to maintain a reasonable weight during pregnancy and give birth at an appropriate age.

Objective To construct a quality control information system for the entire process of neonatal breastfeeding and evaluate its application effect.Methods Based on the"evidence-based guidelines for breastfeeding of inpatient newborns"and combined with clinical practice,hazard analysis and critical control point methods were applied to construct a quality control information system for the entire breastfeeding process,which includes medical order reminder module,knowledge assessment module,breast milk outside the hospital management module,breast milk inventory reminder module,and adverse feeding record module.The pure breastfeeding rate,qualified rate of breast milk quality,and incidence of adverse events of hospitalized newborns before the application of the system(June and August 2021)and after the application of the system(June and August 2022)were compared.Results After the implementation of the system,the pure breastfeeding rate of hospitalized newboms increased from 15.12％to 28.05％,with a statistically significant difference(x2=4.170,P=0.041).The qualified rate of breast milk quality increased from 68.44％to 96.79％,with a statistically significant difference(x2=531.853,P＜0.001).The incidence of adverse events decreased from 5.81％to 0,with a statistically significant difference(P=0.026).Conclusion The quality control information system for the entire process of breastfeeding accurately identified risks and warning signs against them,standardized the management of breast milk outside the hospital,improved the qualification rate of breast milk quality and pure breastfeeding rate,reduced adverse events of breastfeeding,and ensured the effectiveness and safety of breastfeeding.

Objectives:To report a novel mutation site in the pathogenic gene TTR of transthyretin cardiac amyloidosis(ATTR-CA),and to identify family members at risk,and provide suitable clinical diagnosis and treatment. Methods:A retrospective analysis was conducted on the clinical data of the proband with ATTR-CA who visited the Department of Cardiology,the First Affiliated Hospital of Zhengzhou University in March 2021.The proband underwent whole exome sequencing using high-throughput methods to detect mutation genes.Sanger sequencing was used to test candidate pathogenic loci in suspected family members,and relevant literature was reviewed. Results:Among 51 individuals spanning 5 generations in the pedigree,10 family members(including the proband)carried the heterozygous TTR gene c.163A>G mutation,resulting in the amino acid residue at position 55 changing from lysine(Lys)to glutamic acid(Glu).This mutation follows an autosomal dominant inheritance pattern,with early onset in adulthood,rapid progression,and presenting as a mixed-type ATTR-CA.Five mutation carriers had different clinical manifestations,while the remaining 5 mutation carriers,who are at younger age,have not yet shown symptoms.Within the pedigree,7 individuals died(the proband's uncle[Ⅱ-1]who died from stroke at 65 years old,the rest 6 family members died from heart disease before the age of 50). Conclusions:According to the American College of Medical Genetics and Genomics guidelines,TTR gene Lys55Glu mutation is classified as likely pathogenic,this mutation site has not been reported in the literature before.Present study adds clinical evidence that might broaden the spectrum of TTR mutations.

OBJECTIVE To explore the effect of changing lifestyle diet in the treatment of laryngeal reflux disease(LRPD).METHODS A total of 92 patients with LRPD who received outpatient treatment in the Second Affiliated Hospital of Fujian Medical University from January 2022 to June 2023 were selected as the study control,divided into control group(46 cases)and treatment group(46 cases).The control group was treated with conventional acid suppression and gastric motogenic therapy.The treatment group was treated on the basis of the control group with giving guidance on lifestyle and diet.The changes of reflux symptom index scale(RSI)and reflux sign score scale(RFS)and the clinical efficacy of the two groups were compared 8 weeks and 12 weeks after treatment.RESULTS RSI total score was improved 8 weeks after treatment compared with before treatment in both RSI control group and treatment group(P＜0.05).The total RSI score of 12 weeks after treatment was improved compared with that of 8 weeks after treatment in both control and treatment groups(P＜0.05).The total RFS score at 8 weeks after treatment was improved in both control group and treatment group compared with before treatment(P＜0.05).Compared with 8 weeks after treatment,the total RFS score in both control group and treatment group was improved(P＜0.05).After 8 weeks of treatment,the total effective rate of the control group(60.9%)was compared with that of the treatment group(71.7%),and there was no significant difference between the two groups(χ2=1.335,P=0.513).After 12 weeks of treatment,the total effective rate of the control group(73.9%)was compared with the total effective rate of the treatment group(91.3%),and the difference between the two groups was statistically significant(χ2=6.226,P=0.044).CONCLUSION The change of lifestyle and diet should become an important part of the treatment of LRPD.By adjusting the unhealthy lifestyle and diet,the symptoms of patients can be significantly reduced and better clinical efficacy can be obtained.

Objective:To study the current status of longitudinal extrauterine growth restriction (EUGR) in extremely preterm infants (EPIs) and to develop a prediction model based on clinical data from multiple NICUs.Methods:From January 2017 to December 2018, EPIs admitted to 32 NICUs in North China were retrospectively studied. Their general conditions, nutritional support, complications during hospitalization and weight changes were reviewed. Weight loss between birth and discharge > 1SD was defined as longitudinal EUGR. The EPIs were assigned into longitudinal EUGR group and non-EUGR group and their nutritional support and weight changes were compared. The EPIs were randomly assigned into the training dataset and the validation dataset with a ratio of 7∶3. Univariate Cox regression analysis and multiple regression analysis were used in the training dataset to select the independent predictive factors. The best-fitting Nomogram model predicting longitudinal EUGR was established based on Akaike Information Criterion. The model was evaluated for discrimination efficacy, calibration and clinical decision curve analysis.Results:A total of 436 EPIs were included in this study, with a mean gestational age of (26.9±0.9) weeks and a birth weight of (989±171) g. The incidence of longitudinal EUGR was 82.3%(359/436). Seven variables (birth weight Z-score, weight loss, weight growth velocity, the proportion of breast milk ≥75% within 3 d before discharge, invasive mechanical ventilation ≥7 d, maternal antenatal corticosteroids use and bronchopulmonary dysplasia) were selected to establish the prediction model. The area under the receiver operating characteristic curve of the training dataset and the validation dataset were 0.870 (95% CI 0.820-0.920) and 0.879 (95% CI 0.815-0.942), suggesting good discrimination efficacy. The calibration curve indicated a good fit of the model ( P>0.05). The decision curve analysis showed positive net benefits at all thresholds. Conclusions:Currently, EPIs have a high incidence of longitudinal EUGR. The prediction model is helpful for early identification and intervention for EPIs with higher risks of longitudinal EUGR. It is necessary to expand the sample size and conduct prospective studies to optimize and validate the prediction model in the future.

Non-peptide gonadotropin-releasing hormone antagonist(GnRH-antagonist)is a highly efficient and selective antagonist of GnRH receptor.The oral preparations(Elagolix,Relugolix and Linza-golix)had been approved for listing abroad.It shows efficacy in reducing the size of uterine fi-broid and controlling heavy bleeding.Appropriate combination of'add-back'therapy could improve the safety in medical application and patient com-pliance.Compared with other hormonal pharma-ceuticals,oral GnRH-antagonists showed promising prospects in the treatment of uterine fibroid.