An assay was established for detection of toxigenic Clostridioides difficile by combining microfluidic chip analysis with immunochromatography,and its performance was evaluated and compared with those of the Xpert C.difficile/Epi and VIDAS CD AB tests.Primer pairs were designed according to the tcdB and tpi genes in C.difficile.The specificity,limit of detection,reproducibility,and stability were evaluated.A total of 215 stool samples from patients with diarrhea were collected and tested in parallel with the Xpert C.difficile/Epi,VIDAS CDAB,and our assay.C.difficile was isolated from samples,and the tcdB gene was identified when discrepant results were obtained from the three above assays.Our assay showed no cross-reaction with other diarrhea-associated pathogens.Its reproducibility was 100％in testing of two standard plasmids containing tcdB and tpi genes at two concentrations(105 and 102 copies/μL).Two standard plasmids were detected after the PCR and immunochromatography reagents had been stored for 3,6,9,and 12 months,and all the results were posi-tive.The limit of detection was 10 copies/μL for toxigenic C.difficile.Testing of 33 samples positive for C.difficile with our assay(33/215,15.3％)yielded findings statistically coherent with those of the Xpert C.difficile/Epi test(kappa value=0.965).The sensitivity,specificity,positive predictive value,and negative predictive value of our assay,with respect to Xpert C.difficile/Epi as the standard,were 94.3％,100.0％,100.0％,and 98.9％;these values were significantly higher than those of VIDAS CDAB(60.0％,98.9％,91.3％,and 92.7％)(Kappa=0.714,OR=157.50,95％CI:62.03-847.28,P=0.013).In conclusion,our newly developed assay is specific,stable,and reproducible,and may be used for rapid and accu-rate detection of toxigenic C.difficile.The assay could be used for C.difficile infection screening in outpatient and emergen-cy,community medical service center,and epidemiological settings.

Objective To explore the efficacy and safety of 0.02％atropine eye drops for the treatment of nearwork-induced transient myopia(NITM)in adolescents.Methods A total of 131 adolescents with NITM were randomly divided into experimental(receiving 0.02％atropine eye drops)and control(receiving placebo)groups.Changes in the initial NITM values before medication and at 14 and 30 days after medication were observed.Alterations in intraocular pressure and accommodation amplitude were monitored,and the occurrence of complications,such as photophobia and near vision impairment,were recorded.Results The baseline NITM did not differ significantly between the two groups.On day 14 and day 30,the NITM values in the experimental group were significantly reduced compared to the baseline,with differences of 0.31 D±0.20 D and 0.30 D±0.16 D,which were significantly greater than those in the control group(0.21 D±0.98 D and 0.20 D±0.18 D,P＜0.001).The efficacy rate of NITM treatment in the experimental group was 84.4％,which was sig-nificantly higher than that in the control group(29.9％).After 30 days of treatment,no severe systemic or ocular adverse reactions were observed in the experimental group.Mild photophobia was the main adverse reaction.Conclusion 0.02％atropine eye drops can effec-tively reduce the initial NITM value in adolescents within a month of its use,with no severe complications and good tolerance.A clinical trial of atropine eye drops at different concentrations to reduce NITM over a longer period is warranted.

Objective To explore the efficacy and safety of 0.02％atropine eye drops for the treatment of nearwork-induced transient myopia(NITM)in adolescents.Methods A total of 131 adolescents with NITM were randomly divided into experimental(receiving 0.02％atropine eye drops)and control(receiving placebo)groups.Changes in the initial NITM values before medication and at 14 and 30 days after medication were observed.Alterations in intraocular pressure and accommodation amplitude were monitored,and the occurrence of complications,such as photophobia and near vision impairment,were recorded.Results The baseline NITM did not differ significantly between the two groups.On day 14 and day 30,the NITM values in the experimental group were significantly reduced compared to the baseline,with differences of 0.31 D±0.20 D and 0.30 D±0.16 D,which were significantly greater than those in the control group(0.21 D±0.98 D and 0.20 D±0.18 D,P＜0.001).The efficacy rate of NITM treatment in the experimental group was 84.4％,which was sig-nificantly higher than that in the control group(29.9％).After 30 days of treatment,no severe systemic or ocular adverse reactions were observed in the experimental group.Mild photophobia was the main adverse reaction.Conclusion 0.02％atropine eye drops can effec-tively reduce the initial NITM value in adolescents within a month of its use,with no severe complications and good tolerance.A clinical trial of atropine eye drops at different concentrations to reduce NITM over a longer period is warranted.

An efficient analytical method was developed for simultaneous detection of alkylamines and alkylamides in food packaging plastics using liquid chromatography-high resolution mass spectrometry(LC-HRMS).Based on the physicochemical properties of alkylamines and alkylamides,as well as the complexity of plastic samples,sample pretreatment and chromatographic-mass spectrometric parameters were optimized.The samples were extracted by vortex-ultrasonic extraction with a methanol-acetonitrile mixture for 15 min,followed by nitrogen evaporation to concentrate the extract,reconstitution,and analysis.The chromatographic mobile phase consisted of 0.1%formic acid aqueous solution and acetonitrile,and a gradient elution was used.The electrospray ionization(ESI)source was operated in positive ion mode,and mass spectrometry data were collected in full scan and data-dependent acquisition modes.Quantification was performed using an isotope-labeled internal standard method.The results showed that within the quantification range of 1-1000 ng/mL,the calibration curves exhibited good linearity(R2>0.99).Some compounds interfered with the validation experiments at higher concentrations,so only 10 kinds of target analytes were validated.Using a mixed food packaging plastic matrix,the recoveries at spiking levels of 40,400,and 4000 ng/g were mostly between 66.0%and 117.1%,with relative standard deviations ranging from 0.6%to 10.6%.The method was applied to detect 14 food packaging plastic samples,and the results showed that the concentrations of alkylamines and alkylamides ranged from not detected to 8924 ng/g.This method offered high sensitivity and accuracy,and was suitable for the screening and quantitative determination of alkylamines and alkylamides in plastics.

Objective:To evaluate the short-term efficacy of endoscopic-laparoscopic regional gastric resection combined with sentinel lymph node basin dissection in patients with early-stage gastric cancer.Methods:This is a retrospective case series study. Data of 17 consecutive early gastric cancer patients from a prospective cohort at Beijing Friendship Hospital,Capital Medical University were analyzed between August 2023 and August 2024. Sixteen cases were from the department of general surgery and 1 from the department of gastroenterology. The cohort included 9 males and 8 females,with a mean age of 61.4 years (range: 46 to 79 years). Clinical data,including demographics,pathological features,surgical procedures,and follow-up outcomes,were collected through medical records and databases. All patients were followed for over 3 months,with follow-up ending on December 5,2024.Results:A total of 17 patients were involved. Among them, 5 patients underwent endoscopic submucosal dissection (ESD) combined with laparoscopic sentinel lymph node dissection (LSBD),and another 3 patients who underwent complete ESD resection received LSBD due to pathological stage meeting the expanded indications. 6 patients who underwent non-curative ESD resection received laparoscopic gastric regional resection (LRG) combined with LSBD,and another 3 patients directly received LRG combined with LSBD. The average number of sentinel lymph nodes dissected before surgery ( M(IQR)) was 8.9 (4.5) (range: 4 to 21),and the detection rate and accuracy rate were both 100%. Postoperative pathology confirmed that there was no metastasis in the sentinel lymph nodes of 5 patients who underwent ESD combined with LSBD and 3 patients who underwent LSBD after complete ESD resection. The vertical and horizontal margins of ESD were all negative. One patient was an absolute indication for ESD. For the 6 patients who underwent non-curative ESD resection combined with LRG and LSBD,the horizontal margins were all negative. Two patients showed 1 metastasis in each of the 21 and 9 sentinel lymph nodes during the operation,and additional distal gastrectomy was performed during the operation. Postoperatively,73 and 39 lymph nodes were retrieved respectively. The former had 1 additional metastasis,while the latter had no metastasis. Among the 3 patients who underwent direct LRG combined with LSBD,the horizontal margins were negative. One patient was confirmed as an absolute indication for ESD by postoperative pathology,and one patient had 1 metastasis in 8 sentinel lymph nodes during the operation,and additional distal gastrectomy was performed. Postoperatively,there was no metastasis in 54 lymph nodes. All patients had no complications such as infection,bleeding,perforation,or death after surgery. Among the 14 patients who did not receive additional radical surgery,they were able to pass gas and defecate within 3 days after surgery,with an average hospital stay of 6 days. The nutritional indicators and gastric radionuclide emptying imaging half-emptying time were similar to those before surgery at 3 months after surgery. Conclusions:Laparoscopic and endoscopic cooperative regional gastrectomy with sentinel lymph node basin dissection has the advantages of minimal invasiveness,preservation of gastric function,and precise treatment. It maybe suitable for patients with early-stage gastric cancer at high risk of lymph node metastasis and has good short-term efficacy.

An assay was established for detection of toxigenic Clostridioides difficile by combining microfluidic chip analysis with immunochromatography,and its performance was evaluated and compared with those of the Xpert C.difficile/Epi and VIDAS CD AB tests.Primer pairs were designed according to the tcdB and tpi genes in C.difficile.The specificity,limit of detection,reproducibility,and stability were evaluated.A total of 215 stool samples from patients with diarrhea were collected and tested in parallel with the Xpert C.difficile/Epi,VIDAS CDAB,and our assay.C.difficile was isolated from samples,and the tcdB gene was identified when discrepant results were obtained from the three above assays.Our assay showed no cross-reaction with other diarrhea-associated pathogens.Its reproducibility was 100％in testing of two standard plasmids containing tcdB and tpi genes at two concentrations(105 and 102 copies/μL).Two standard plasmids were detected after the PCR and immunochromatography reagents had been stored for 3,6,9,and 12 months,and all the results were posi-tive.The limit of detection was 10 copies/μL for toxigenic C.difficile.Testing of 33 samples positive for C.difficile with our assay(33/215,15.3％)yielded findings statistically coherent with those of the Xpert C.difficile/Epi test(kappa value=0.965).The sensitivity,specificity,positive predictive value,and negative predictive value of our assay,with respect to Xpert C.difficile/Epi as the standard,were 94.3％,100.0％,100.0％,and 98.9％;these values were significantly higher than those of VIDAS CDAB(60.0％,98.9％,91.3％,and 92.7％)(Kappa=0.714,OR=157.50,95％CI:62.03-847.28,P=0.013).In conclusion,our newly developed assay is specific,stable,and reproducible,and may be used for rapid and accu-rate detection of toxigenic C.difficile.The assay could be used for C.difficile infection screening in outpatient and emergen-cy,community medical service center,and epidemiological settings.

Objective:To evaluate the short-term efficacy of endoscopic-laparoscopic regional gastric resection combined with sentinel lymph node basin dissection in patients with early-stage gastric cancer.Methods:This is a retrospective case series study. Data of 17 consecutive early gastric cancer patients from a prospective cohort at Beijing Friendship Hospital,Capital Medical University were analyzed between August 2023 and August 2024. Sixteen cases were from the department of general surgery and 1 from the department of gastroenterology. The cohort included 9 males and 8 females,with a mean age of 61.4 years (range: 46 to 79 years). Clinical data,including demographics,pathological features,surgical procedures,and follow-up outcomes,were collected through medical records and databases. All patients were followed for over 3 months,with follow-up ending on December 5,2024.Results:A total of 17 patients were involved. Among them, 5 patients underwent endoscopic submucosal dissection (ESD) combined with laparoscopic sentinel lymph node dissection (LSBD),and another 3 patients who underwent complete ESD resection received LSBD due to pathological stage meeting the expanded indications. 6 patients who underwent non-curative ESD resection received laparoscopic gastric regional resection (LRG) combined with LSBD,and another 3 patients directly received LRG combined with LSBD. The average number of sentinel lymph nodes dissected before surgery ( M(IQR)) was 8.9 (4.5) (range: 4 to 21),and the detection rate and accuracy rate were both 100%. Postoperative pathology confirmed that there was no metastasis in the sentinel lymph nodes of 5 patients who underwent ESD combined with LSBD and 3 patients who underwent LSBD after complete ESD resection. The vertical and horizontal margins of ESD were all negative. One patient was an absolute indication for ESD. For the 6 patients who underwent non-curative ESD resection combined with LRG and LSBD,the horizontal margins were all negative. Two patients showed 1 metastasis in each of the 21 and 9 sentinel lymph nodes during the operation,and additional distal gastrectomy was performed during the operation. Postoperatively,73 and 39 lymph nodes were retrieved respectively. The former had 1 additional metastasis,while the latter had no metastasis. Among the 3 patients who underwent direct LRG combined with LSBD,the horizontal margins were negative. One patient was confirmed as an absolute indication for ESD by postoperative pathology,and one patient had 1 metastasis in 8 sentinel lymph nodes during the operation,and additional distal gastrectomy was performed. Postoperatively,there was no metastasis in 54 lymph nodes. All patients had no complications such as infection,bleeding,perforation,or death after surgery. Among the 14 patients who did not receive additional radical surgery,they were able to pass gas and defecate within 3 days after surgery,with an average hospital stay of 6 days. The nutritional indicators and gastric radionuclide emptying imaging half-emptying time were similar to those before surgery at 3 months after surgery. Conclusions:Laparoscopic and endoscopic cooperative regional gastrectomy with sentinel lymph node basin dissection has the advantages of minimal invasiveness,preservation of gastric function,and precise treatment. It maybe suitable for patients with early-stage gastric cancer at high risk of lymph node metastasis and has good short-term efficacy.

Objective To explore the clinical characteristics and treatment scheme of patients with spinal infection caused by Prevotella intermedia(P.intermedia).Methods Clinical diagnosis and treatment processes of a patient with spinal infection caused by P.intermedia admitted to the spinal surgery department of a hospital were summa-rized,and relevant literature was retrieved from database for reviewing.Results The patient,a 50 year old male,was admitted to the hospital due to"lumbago pain complicated with pain in double lower extremities for 2 months".The lesion tissue was taken for metagenomic next-generation sequencing(mNGS)detection,which detected P.in-termedia,and the patient was diagnosed with P.intermedia spondylitis.After treatments with open lesion clea-rance,tube rinsing+autologous bone transplantation fusion internal fixation,intravenous drip of ceftriaxone sodium and metronidazole,as well as metronidazole rinsing,infection was under control.A total of 16 available papers were retrieved,together with this case,a total of 17 patients were included,with 7 males and 10 females.The main risk factors were diabetes and history of corticosteroid use(35.3％).The most common invasion sites were lumbar ver-tebra(n=12)and thoracic vertebra(n=6).13 cases were positive for pathogen culture,3 cases were positive for molecular detection,and 1 case was positive for staining microscopy.17 patients received anti-anaerobic bacteria treatment,with 14 cases receiving combined surgical treatment.One case died,with a mortality of 5.9％;5 cases had partial neurological impairment,with a disability rate of 29.4％.The survival rate of patients who received treatment of anti-anaerobic bacteria combined with surgery was 92.8％,3 patients only with anti-anaerobic bacteria treatment but without surgery were all cured.Conclusion P.intermedia is an opportunistic pathogeanic bacteria which often causes infection in immunocomprised individuals and is prone to be misdiagnosed.It is recommended to perform mNGS detection to identify the pathogen as early as possible and seize the opportunity for treatment to reduce mortality.

Objective:To analyze the relationship between serum cystatin C(CysC),β2-microglobulin(β2-MG)and the efficacy of demethylation therapy in patients with acute myeloid leukemia(AML).Methods:A prospective cohort study was conducted on 98 AML patients admitted to the Affiliated Hospital of Inner Mongolia Medical University from February 2019 to January 2022.All patients were treated with decitabine(DAC)+HAG regimen,28 days as a course and treated for 3-4 courses.At the end of each course of treatment,the treatment effect of the patients was evaluated,and the patients who achieved complete remission(CR)transferred to consolidation therapy,while the patients who did not reach CR at the end of the course of treatment were considered as treatment failure.The examination items before treatment include routine blood parameters,serum CysC,and β2-MG,and general clinical data of the patients were collected.According to the statistical results,logistic regression model was used to analyze the relationship between serum CysC,β2-MG and the efficacy of demethylation therapy in AML patients.The ROC curves were drawn,and the predictive efficacy of serum CysC,β2-MG on demethylation therapy in AML patients was evaluated by the area under the curve(AUC).Results:Of the 98 AML patients enrolled in the study,5 cases were excluded during the treatment period,and 93 cases finally completed the chemotherapy courses.Among them,23 patients achieved CR after the initial induction chemotherapy(course 1-2),and 11 patients achieved CR after the re-induction chemotherapy(course 3-4).The success rate of demethylation therapy was 36.56％(34/93).Compared with the patients in treatment success group,patients in treatment failure group had a higher proportion of intermediate-and adverse-risk,lower levels of platelet(PLT)and hemoglobin(Hb),and higher expression levels of serum CysC and β2-MG,all of which were statistically significant(P＜0.05).Logistic regression analysis showed that high expression of serum CysC,β2-MG and adverse-risk were independent risk factors for failure of demethylation treatment in AML patients(OR＞1,P＜0.05).The ROC curves showed that the AUC values of serum CysC,β2-MG alone and combined in predicting the efficacy of demethylation therapy in AML patients were 0.788,0.785 and 0.834,respectively.Conclusion:The failure of demethylation therapy in AML patients is related to the high expression of serum CysC and β2-MG,and detection of serum CysC and β2-MG before treatment can predict the risk of demethylation therapy failure in AML patients.

Objective To explore the feasibility and rationality of advanced integrated two-stage laparoscopic simulation training course in standardized training of surgical residents.Methods From December 2019 to December 2021,the advanced integrated two-stage laparoscopic simulation training course was carried out among 2019-2020 surgical residents who received standardized training in our hospital.The course was divided into two stages.In the first stage,BEST(best essential surgical technology training)course,adopted Darwin? endoscopic training system,Tianyan? endoscopic training system,Microport? 3D laparoscopic training system and simple simulative models were used.The second stage,BEST PLUS course,same platform as that in BEST course and in vitro animal models were used.The questionnaire survey method(before and after class questionnaire)was adopted to evaluate the curriculum setting,such as curriculum form,simulators,teaching method,time arrangement,curriculum difficulty,training effect,curriculum satisfaction and so on.Results A total of 37 surgical residents completed the two-stage course training and the questionnaire survey.The overall satisfaction rate with the curriculum setting was 100%.There were 32 residents(86.5%)thought that first stage training course could significantly improve their clinical skills,35 residents(94.6%)thought that second stage training course could significantly improve their clinical skills,and 36 resident(97.3%)thought that the first stage curriculum could significantly help them improve performance in the second stage curriculum.Conclusions The trainees had a high degree of recognition and satisfaction for the advanced integrated two-stage laparoscopic simulation training course.The overall design of course was reasonable and feasible,and was attractive to trainees.