Objective: To establish a rapid GeXP-based multiplex reverse transcription-PCR assay (GeXP assay) for simultaneous detection of 5 subtypes of diarrheogenic Escherichia coli. Methods: Specific primers were designed according to reserved sequences of 12 virulence genes in enterotoxigenic E. coli (ETEC), enterinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC) and enterohemorrhagic E. coli (ETEC), and PCR amplification was performed with a single pair of primers to validate the specificity of PCR assay with a single template and a single pair of primers. The specificity of the GeXP assay was evaluated with the genomic DNA of 5 subtypes of diarrheogenic E. coli as the template in a mixture of 12 pairs of primers, and the sensitivity of the GeXP assay was evaluated with the mixed suspensions of 5 subtypes of diarrheogenic E. coli at concentrations of 106, 105, 104 and 103 CFU/mL as the template. Foods purchased from supermarkets and agricultural retail markets were prepared into 34 spiked samples, and 5 subtypes of diarrheogenic E. coli were detected using the GeXP assay and compared with the fluorescent real-time multiple PCR assay. Results: The sizes of sth, pic, bfpB, astA, lt, escV, aggR, stx1, uidA, invE, stx2 and stp genes amplification products were consistent with expected sizes using a single template and a single pair of primers, with a fluorescent signal intensity of more than 25 000 A.U. The sizes of the GeXP assay amplification products of 12 virulence genes in 5 subtypes of diarrheogenic E. coli were consistent with expected sizes, with a high specificity. If the concentration of the mixed suspensions of 5 subtypes of diarrheogenic E. coli was 103 CFU/mL, the GeXP assay was effective for simultaneous detection of 12 virulence genes, with a high fluorescent signal intensity, consistent repeated detection results and a less than 10% coefficient of variation. The GeXP assay detected 3 ETEC isolates, 12 EAEC isolates, one EIEC isolate, one EPEC isolate and one EHEC isolate among the 34 spiked samples, which was in agreement with the detection of 5 subtypes of diarrheogenic E. coli with commercial fluorescent real-time multiple PCR assay kits. Conclusions A GeXP assay has been successfully established for simultaneous detection of 12 virulence genes in diarrheogenic E. coli, which is effective for clinical differential diagnosis and epidemiological surveys of diarrheogenic E. coli.

Objective: To investigate the contamination status and assess the potential consumption risk of Listeria monocytogenes ( L. monocytogenes ) in cooked meat products in bulk in Zhejiang Province, so as to provide strategy for food safety supervision and management. Methods: A total of 2 320 cooked meat products were sampled from eleven cities in Zhejiang Province during 2018-2020. The detection of L. monocytogenes was carried out in accordance with the national standard GB/T 4789.30-2016. Risk Ranger software was used for the semi-quantitative risk assessment on the whole population and pregnant women. Results: The total detection rate of L. monocytogenes in cooked meat products in bulk in Zhejiang Province was 2.97% ( 69/2 320 ). The detection rates in stewed, smoked/roasted, fried, dried products and others were 3.85%, 1.81%, 0.59%, 0% and 0.94%, which were significantly different ( P<0.05 ). There were 28 positive samples in 1 069 samples collected in 2020, with the concentration ranging from 5 to 590 CFU/g and averaging 6.8 CFU/g. The estimated number of listeriosis cases each year caused by consumption of cooked meat products in bulk was 131 in the whole population with a risk score of 42, and 1.44 in pregnant women with a risk score of 54. The risk coefficient could reduce to approximate zero after sufficient heating before intake. Conclusion The prevalence of L. monocytogenes in cooked meat products in bulk in Zhejiang Province during 2018-2020 poses a potential risk in food safety. Pregnant women should avoid eating.

Objective: To establish real-time recombinase polymerase amplification（RPA）for the rapid detection of Vibrio parahaemolyticus（VP）. Methods: An exo probe and primers were designed according to the conserved sequence of thermolabile hemolysin（tlh）gene of VP and then RPA for detection of VP was established. The sensitivity of the assay was evaluated by detecting different concentration of VP;the specificity was evaluated by detecting different bacteria;the stability was evaluated by repeat trials;the application effect was evaluated by detecting food samples which were simultaneously tested with traditional culture method according to GB 4798.7-2013 Detection of VP. Results: A real-time RPA was established to complete VP amplification within 20 min at a constant temperature of 39 ℃. The analytical sensitivity of the assay was five pg per reaction and no cross-reactivity with other pathogenic bacteria observed. The RPA detection results with different concentration of VP and E. coli DNA templates at three time points were consistent. The detection results of 51 food samples by RPA were the same as those by traditional culture method. Conclusion The established real-time RPA can qualitatively detect VP,with simple operation and interpretation of results,which is suitable for rapid detection of VP in public health emergencies and food safety supervision.

OBJECTIVETo establish the differential templates of the skeletal profiles for Shanghainese with normal occlusion in early permanent dentition.

METHODS71 lateral cephalometric radiographs of subjects with normal occlusion in early permanent dentition, aged from 11 - 15 years (37 females and 34 males) were traced. 23 skeletal landmarks were identified on the tracings, which afterwards were scanned on the computer. An x-y coordinate system was established: the tracing superimposed on sella rotated 7 degrees down from the sella-nasion line as the x axis and the vertical line through sella perpendicular to the x axis as the y axis. A special length/depth ratio was designed. The length/depth ratio of each graph were calculated. Steiner analysis was applied to check the difference between genders.

RESULTSThere was no difference between genders. The tracings were divided into three categories: short face, normal face and long face according to the mean and standard deviation of the ratio. The three types of tracings were superimposed on the x axis with sella registered. Three templates were created.

CONCLUSIONSThe differences were apparent when the three templates were superimposed.

Adolescent ; Cephalometry ; methods ; Child ; Dental Occlusion ; Face ; anatomy & histology ; Facial Bones ; anatomy & histology ; Female ; Humans ; Male

Objective:To summarize our experience in prevention of COVID-19 infection in emergency and confined operations during the first 3 weeks after Spring Festival in 2020.Methods:From February 3rd to 23rd, 2020, 151 patients were admitted to Department of Orthopaedic Trauma, Beijing Jishuitan Hospital for emergency and confined operations. In this cohort, 125 patients were admitted to ordinary wards. They were 70 males and 55 females with an age of 51.1 years ± 14.9 years. Of them, 2 were subjected to emergency operation and 123 to confined operation. The mean time from injury to operation was 9.9 days ± 6.1 days. There were 26 cases in the senile wards, 7 males and 19 females with an age of 80.8 years ± 7.0 years all of whom underwent confined operations. The mean time from injury to operation was 8.4 days ± 6.3 days. The protocols for emergency diagnosis, admission, emergency and confined operations, postoperative rehabilitation and management of suspects with COVID-19 during the epidemic of COVID-19 were optimized according to Diagnosis and Treatment Protocols for Novel Coronavirus Pneumonia (Trial version 5), emergency responding pre-plans of our hospital, and our experience in Enhanced Recovery After Surgery (ERAS) as well.Results:The patients in the ordinary wards had a hospital stay of 6.8 days ± 4.6 days while those in the senile wards 5.1 days ± 2.0 days. Abnormal temperature (≥37.3 ℃) was observed perioperatively in 17 cases in the ordinary wards. It was absorption fever in all and appeared in 4 cases upon admission. Fever appeared in 11 patients in the senile wards and upon admission in 3 of them. One senile patient who had been diagnosed of normal pneumonia returned to normal temperature and remained stable conditions after antibiotic therapy. The other patients were free of complications related to COVID-19 during their hospital stay.Conclusion:The first-line medical staff working at emergency department, wards and surgical theaters must heighten their vigilance against COVID-19 infection and rigorously follow protocols for prevention of COVID-19 infection in their daily clinical practice.

Objective:To elucidate the recent therapeutic efficacy of the intelligent fracture reduction robotic system in managing pelvic fractures.Methods:A retrospective evaluation of 49 pelvic fracture patients treated using the intelligent fracture reduction robotic system at Beijing Jishuitan Hospital's trauma orthopedics department between March 2021 and December 2022 was conducted. The cohort included 30 males and 19 females, with a mean age of 51.51±18.71 years (20-92 years range). Fractures were classified according to the Tile system: B1 type in 2 cases, B2 in 7, B3 in 3, C1 in 30, and C2 in 3. The median interval between injury and surgery was 6 days, with a range of 2-22 days. The robotic system assisted in pelvic fracture reduction and stabilization surgeries. Preoperative and postoperative evaluations involved pelvic CT scans, anteroposterior, inlet, and outlet radiographic images. Fracture displacement and reduction outcomes were assessed via X-ray imagery. Data captured included intraoperative blood loss, duration of surgery, fracture stabilization techniques, and postoperative monitoring period. The Majeed scoring system gauged functional outcomes.Results:Of the patients, 48 underwent minimally invasive interventions with robotic assistance, while one case necessitated open reduction and internal fixation due to an unsuccessful reduction. The duration between injury and operation ranged from 2 to 22 days. Average surgical time stood at 206.5±7.1 minutes (105-440 min range), and median intraoperative blood loss was 100ml (10-600 ml range). Using the Matta reduction criteria, 30 postoperative cases exhibited excellent and 9 good outcomes for posterior pelvic ring displacement, translating to a 93% (38/41) positive rate. For anterior pelvic ring shifts, 45 showed excellent and 3 good outcomes, culminating in a 100% (48/48) success rate. Follow-up for the 48 cases lasted 11.0 months (3-23 months range), with the Majeed functional score averaging 81.9±17.0 points (42-100 point range). 27 cases scored excellent, and 11 good, yielding a combined positive outcome rate of 79.2% (38/48).Conclusion:Employing the intelligent fracture reduction robotic system in pelvic fracture treatments facilitates minimally invasive interventions and yields favorable short-term clinical results.

Objective: To explore age-related clinical characteristics, early responses and outcomes in non-senile adults with de novo acute myeloid leukemia (AML). Methods: Data of consecutive cases of 18-65 years adults with de novo AML (non-acute promyelocytic leukemia) were reviewed retrospectively. Clinical characteristics at diagnosis, early responses and outcomes across different age groups of patients were analyzed. Results: 1 097 patients were enrolled. 591 (53.9%) were male. Median age was 42 years. Increasing age was significantly associated with decreasing WBC count (P=0.003), increasing PLT count (P=0.034), lower blast proportions in bone marrow (P=0.021). The incidence of NPM1⁺/FLT3-ITD^- increased with age (P<0.001). Multivariate analyses showed that increasing age was associated with low probabilities of achieving morphologic leukemia free state (MLFS) (P=0.053) and complete remission (CR) (P=0.004) and poor overall survival (OS) (P=0.070) in the whole patients population. However, increasing age was not associated with low MLFS rate and poor OS, except low CR rate (P=0.075) in those receiving standard induction regimen instead of low-intensity regimen. Conclusions There were significant differences on clinical characteristics, cytogenetics and molecular genetics across different age groups in non-senile adults with de novo AML. In the patients receiving standard induction regimen, age was not associated with MLFS rate and OS.

Objective: To explore prognostic significance of early assessment of minimal residual leukemia (MRD) in adult patients with de novo acute myeloid leukemia (AML) with mutated NPM1. Methods: The response, NPM1 mutated transcript level after induction chemotherapy and the first 2 cycles of consolidation chemotherapy, disease-free survival (DFS) and overall survival (OS) in 137 patients with AML with NPM1 mutations of A, B and D were retrospectively analyzed. Results: Data of 137 patients were collected, 67 were male, the median age was 49 years (16-67 years) , 107 (78.1%) had normal karyotype, 57 (41.6%) had positive FLT3-ITD mutation, the median NPM1 mutated transcript level at diagnosis was 84.1%. Among the 134 evaluable patients, 115 (85.8%) achieved a complete remission (CR) . Multivariate analyses revealed that WBC<100×10⁹/L (OR=0.3, 95% CI 0.1-0.9, P=0.027) and first induction therapy with "IA10" protocol (OR=0.3, 95% CI 0.1-0.8, P=0.015) were factors associated with achieving a CR. With a median follow-up period of 24 months (range, 2 to 91 months) in 77 survived CR patients, the probabilities of DFS and OS at 3 years were 48.0% and 63.9%, respectively. Multivariate analyses showed that positive FLT3-ITD (HR=3.2, 95% CI 1.6-6.7, P=0.002) , high MRD level after 2 cycles of consolidation chemotherapy (NPM1 mutation transcript level <3-log reduction from the individual baseline, HR=23.2, 95% CI 7.0-76.6, P<0.001) and chemotherapy or autologous hematopoietic stem cell transplantation (auto-HSCT) rather than allogeneic HSCT (allo-HSCT) (HR=2.6, 95% CI 1.0-6.6, P=0.045) were the unfavorable factors affecting DFS, high MRD level at the time of achieving the first CR (NPM1 mutation transcript level <2-log reduction from the individual baseline, OR=2.5, 95% CI 1.0-6.1, P=0.040) and after 2 cycles of consolidation chemotherapy (HR=4.5, 95% CI 2.0-10.3, P<0.001) were the unfavorable factors affecting OS. Furthermore, DFS and OS rates at 3 years in those receiving chemotherapy or auto-HSCT were 39.7% and 59.1%, respectively; positive FLT3-ITD and high MRD level after 2 cycles of consolidation chemotherapy were independent factors associated with both shorter DFS (HR=3.5, 95% CI 1.6-7.6, P=0.002 and HR=8.9, 95% CI 3.8-20.7, P<0.001) and OS (HR=2.7, 95% CI 1.1-6.9, P=0.036 and HR=3.1, 95% CI 1.2-8.0, P=0.021) ; meanwhile, high MRD level at the time of achieving the first CR associated with shorter OS (HR=3.1, 95% CI 1.2-8.0, P=0.022) . Conclusion Positive FLT3-ITD mutation and high MRD level after induction or consolidation chemotherapy associated with poor outcomes in AML patients with mutated NPM1.

OBJECTIVE: To explore the clinical characteristics and outcomes of adult critically ill patients with COVID-19 and identify the risk factors correlated with in-hospital deaths. METHODS: This study was conducted among 20 confirmed adult cases of COVID-19 in the Intensive Care Unit (ICU) of Honghu People's Hospital in Jingzhou City, Hubei Province. According to the final outcome, the patients were divided into survivor group and death group with 10 patients each. The demographic data, clinical manifestations and signs, laboratory findings, treatment measures and clinical outcomes were obtained from electronic medical records to compare the clinical characteristics and outcomes between the two groups. Univariate logistic analysis was used to analyze the risk factors associated with in-hospital death. RESULTS: The mean age of patients with confirmed COVID-19 was 70 ± 12 years, and 40% of them were male. The patients were admitted to ICU 11 ± 9 days after symptom onset. The most common symptoms on admission were cough (19 cases), fatigue or myalgia (18 cases), fever (17 cases) and dyspnea (16 cases). Eleven (55%) of the patients had underlying diseases, among which hypertension was the most common (11 cases), followed by cardiovascular disease (4 cases) and diabetes (3 cases). Six (30%) of the patients received invasive mechanical ventilation and continued renal replacement therapy but eventually died. Acute cardiac injury was the most common complication (19 cases). Half of the patients died between the 2nd and 19th day after ICU admission. Compared with dead patients, the surviving patients had a lower average body weight (61.70±2.36 68.60±7.15 kg, =0.01) and a higher Glasgow Coma Index (14.69 ± 0.70 12.70 ± 2.45, =0.03), and were less likely to develop shock (2 10, =0.001) or acute respiratory distress syndrome (2 10, =0.001). CONCLUSIONS Critically ill patients with COVID-19 are generally older. A higher body weight and a lower lymphocyte count are potentially associated with a greater likeliness of fatality in ICU patients with COVID-19.
Aged ; Aged, 80 and over ; Betacoronavirus ; Coronavirus Infections ; Critical Illness ; Female ; Humans ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral ; Retrospective Studies

OBJECTIVETo compare the outcomes of patients with adult acute lymphoblastic leukemia (ALL) over the last 14 years by taking 2006 as the demarcation point.

METHODSFrom January 2000 to December 2013, 477 consecutive hospitalized patients with adult ALL were retrospectively analyzed, including 276 (57.9%) with Ph negative B-ALL (Ph⁻-B-ALL) B-ALL, 69 (14.5%) with T-ALL and 132 (27.7%) with Ph positive ALL (Ph⁺-ALL); 111 (23.3%) before 2006 and 366 (76.7%) after 2006. Among 435 patients who achieved complete remission (CR), 248 (57.0%) received allogeneic hematopoietic stem cell transplantation (allo-HSCT), and 187 remained on chemotherapy.

RESULTSWith a median follow-up period of 19 months in all patients and 35 months in living patients, overall CR rate was 92.0%. Of 435 CR patients, the cumulative incidences of 5-year relapse, disease-free survival( DFS) and overall survival (OS) rates were 42.5%, 46.2% and 47.6%, respectively. Compared with the patients hospitalized before 2006: ① the Ph+-ALL patients hospitalized after 2006 achieved a higher overall CR rate (P=0.036). There was no difference for CR rates of Ph--B-ALL and T-ALL patients between before and after 2006. ②The CR patients hospitalized after 2006 had higher 5-year DFS and OS rates (P=0.001; P < 0.001), including higher 5-year OS rate in Ph⁻-B-ALL patients (P=0.046), and higher 5-year DFS and OS rates in both T-ALL (P=0.013; P=0.036) and Ph⁺-ALL patients (P=0.003; P < 0.001) , especially in those Ph⁺-ALL patients who received imatinib from the beginning of the induction chemotherapy (P < 0.001; P < 0.001) ③The patients who received allo-HSCT after 2006 had higher 5-year DFS and OS rates (P=0.001; P < 0.001), but there was no difference for the outcomes in those who remained on chemotherapy before and after 2006. After 2006, Ph⁺-ALL patients who received imatinib from the beginning of the induction chemotherapy had the highest 5-year DFS and OS rates compared with Ph⁻-B-ALL and T-ALL patients (P=0.009; P=0.001) . Multivariate analysis showed that allo-HSCT and imatinib were two important factors affecting the outcomes of ALL patients.

CONCLUSIONThe outcomes of ALL patients improved significantly over the last 14 years, especially in Ph⁺-ALL ones.

Acute Disease ; Adult ; Disease-Free Survival ; Hematopoietic Stem Cell Transplantation ; Humans ; Imatinib Mesylate ; therapeutic use ; Induction Chemotherapy ; Multivariate Analysis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; therapy ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; therapy ; Recurrence ; Remission Induction ; Retrospective Studies ; Survival Rate