AIM:This study aims to explore the impact of Escherichia coli(E.coli)high-pathogenicity island(HPI)on pyroptosis and intestinal inflammation.METHODS:Kunming mice and IPEC-J2 cells(porcine small intesti-nal epithelial cells)were treated with HPI-containing E.coli strain(HPI+),HPI-deleting E.coli strain(ΔHPI),or lipo-polysaccharide(LPS).The intestinal lactate dehydrogenase(LDH)activity,superoxide dismutase(SOD)activity,IgA expression and secretory IgA(SIgA)content were assessed in mice.The expression of key regulatory factors in the nucleo-tide-binding oligomerization domain-like receptor protein 3(NLRP3)/caspase-1 signaling pathway-related proteins in mouse intestinal tract and IPEC-J2 cells was analyzed by RT-qPCR,immunohistochemical staining and Western blot.The levels of interleukin-1β(IL-1β)and IL-18 in mouse serum and IPEC-J2 cell culture supernatants were measured by ELISA.The pivotal role of NLRP3 in HPI+infection was confirmed by silencing NLRP3 in IPEC-J2 cells using siRNA.RE-SULTS:The HPI+infection markedly decreased SOD activity,increased IgA+B cell count,and induced the LDH release and SIgA secretion in the mouse intestine compared with ΔHPI infection.The results of ELISA,HE staining and TUNEL staining indicated that E.coli HPI triggered DNA damage,tissue injury and inflammation in mouse intestinal epithelial cells.Western blot revealed an increase in intestinal gasdermin D N-terminal fragment(GSDMD-N)protein level with HPI+infection compared with ΔHPI infection.E.coli HPI significantly up-regulated mRNA and protein expression of NLRP3,apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),caspase-1,GSDMD,IL-1β and IL-18 in mouse intestinal tissues and IPEC-J2 cells,accompanied by the elevated secretion of IL-1β and IL-18.The confo-cal microscopy demonstrated an enhanced assembly of the NLRP3 inflammasome with HPI+infection compared with ΔHPI infection,leading to colocalization of NLRP3 and caspase-1.Furthermore,NLRP3 silencing in IPEC-J2 cells attenuated E.coli HPI-induced cell inflammation,damage,and NLRP3/caspase-1 signaling pathway activation.CONCLUSION:The presence of HPI enhances the virulence of E.coli and exacerbates intestinal inflammation.Moreover,pyroptosis,regu-lated by the NLRP3/caspase-1 signaling pathway,plays a pivotal role in the intestinal injury induced by E.coli HPI.

Background/Aims: Enteroenteric intussusception in Peutz-Jeghers syndrome (EI-PJS) is traditionally treated by surgery. However, enteroscopic treatment is a minimally invasive approach worth attempting. We aimed to develop a risk scoring system to facilitate decision-making in the treatment of EI-PJS. Methods: This was a single-center case-control study, including 80 patients diagnosed with PJS and coexisting intussusception between January 2015 and January 2021 in Air Force Medical Center. We performed logistic regression analysis to identify independent risk factors and allocated different points to each subcategory of risk factors; the total score of individuals ranged from 0 to 9 points. Then, we constructed a risk stratification system based on the possibility of requiring surgery: 0–3 points for “low-risk,” 4–6 points for “moderate-risk,” and 7–9 points for “high-risk.” Results: Sixty-one patients (76.25%) were successfully treated with enteroscopy. Sixteen patients (20.0%) failed enteroscopic treatment and subsequently underwent surgery, and three patients (3.75%) received surgery directly. Abdominal pain, the diameter of the responsible polyp, and the length of intussusception were independent risk factors for predicting the possibility of requiring surgery. According to the risk scoring system, the incidence rates of surgery were 4.44% in the low-risk tier, 30.43% in the moderate-risk tier, and 83.33% in the high-risk tier. From low- to high-risk tiers, the trend of increasing risk was significant (p<0.001). Conclusions We developed a risk scoring system based on abdominal pain, diameter of the responsible polyps, and length of intussusception. It can preoperatively stratify patients according to the risk of requiring surgery for EI-PJS to facilitate treatment decision-making.

Objective:To design and construct a bone nonunion organoid on chip and explore the mechanism of aseptic bone nonunion.Methods:First a semi-open microfluidic chip was designed, on which human bone marrow mesenchymal stromal cells (BMSC), human fetal lung fibroblast 1, (HFL1) and human umbilical vein endothelial cells (HUVEC) were co-cultured, and a three-dimensional organ on chip system was established. Different proportions of HFL1 and HUVEC were co-cultured with BMSC, which were divided into the control group (HFL1∶HUVEC=1∶1), the fibrosis group (HFL1∶HUVEC=3∶1) and the vascularization group (HFL1∶HUVEC=1∶3). The osteogenic differentiation of BMSC was observed by alkaline phosphatase (ALP) and Alizarin red staining. The transcription level of osteogenic marker genes SP7, RUNX2, ALPL, and BGLAP, and vascularization related genes KDR and VWF were analyzed by qPCR. The expression levels of RUNX2 and ALP were determined by Western Blot. Results:In the co-culture system of BMSCs, HFL1, and HUVECs, BMSCs exhibited normal growth and apparent biomineralization behavior. Endothelial cells were capable of forming structured vascular networks, confirming the successful establishment of the system. Compared to the baseline group, the fibrotic group showed no significant decrease in BMSC osteogenic differentiation. The relative expression levels of the mineralization marker genes ALPL and BGLAP were 0.55±0.19 ( P<0.001) and 0.42±0.27 ( P<0.001), respectively. Vascularization genes KDR and VWF were downregulated, with relative expression levels of 0.49±0.17 ( P<0.001) and 0.49±0.21 ( P<0.001). In contrast, in the vascularized group, BMSC osteogenic differentiation genes SP7, RUNX2, ALPL, and BGLAP were upregulated, with relative expression levels of 2.91±0.52 ( P<0.001), 3.83±1.87 ( P<0.001), 3.22±1.29 ( P<0.001), and 5.21±1.46 ( P<0.001), respectively. Vascularization genes KDR and VWF were also upregulated, with relative expressions of 8.24±2.84 ( P<0.001) and 5.32±1.67 ( P<0.001). Western blot results indicated increased expression of RUNX2 and ALP in the vascularized group and decreased expression in the fibrotic group. Conclusion:The bone nonunion organoid on chip could partially simulate the local microenvironment of bone nonunion. Fibrosis may lead to a significant decrease in bone formation ability and vascularization level, which might be an important reason for the occurrence of aseptic bone nonunion.

Objective:To evaluate the effectiveness and safety of ultrasound-guided hydrostatic reduction for pediatric acute intussusception.Methods:One thousand eight hundred and thirty patients with acute intussusception diagnosed by ultrasound in Shenzhen Children′s Hospital from September 2017 to July 2020 were treated with ultrasound-guided hydrostatic reduction method. The therapeutic effects, complications and ultrasonic features were observed.Results:Among 1 830 cases, 1 791 cases were diagnosed as primary intussusception, and 39 cases were secondary intussusception. The overall rate of successful ultrasound enema reduction were 1 780/1 830(93.7%) patients. All 50/1 830(2.7%) patients underwent surgery after unsuccessful enema reduction, including 42 cases of primary intussusception, and 8 cases of secondary intussusception. The complication of intestinal perforation occurred in 3 cases (0.16%), and there were no deaths.Conclusions:Ultrasound-guided enema reduction for pediatric acute intussusception is an effective and safe method without radiation exposure, and can be used as the preferred method for non-operative treatment of intussusception.

Objective:To evaluate the effect of general anesthesia on microelectrode recording (MER) during deep brain stimulation (DBS) of subthalamic nucleus (STN) in the patients with primary Parkinson′s disease (PD).Methods:Forty-four patients of both sexes with primary PD (duration of disease ≥ 5 yr and/or obvious symptom fluctuation), undergoing bilateral STN DBS from March 2008 to March 2018, aged<80 yr, were selected and divided into 2 groups by a random number table method: awake group ( n=26) and general anesthesia group ( n=18). In awake group, 0.5% ropivacaine was used for incision infiltration at skin incision.Patients in GA group received propofol and remifentanil by target-controlled infusion with Narcotrend to monitor the depth of anesthesia, and 0.5% ropivacaine was used for incision infiltration at skin incision.The total number of trajectories and length of STN were recorded during MER.Movement disorders were evaluated at 1 week before surgery and 6 months after surgery, and the improvement rate of dyskinesia was calculated.The postoperative anesthesia-, hardware- and stimulation-related complications were recorded. Results:There were no significant differences between the two groups in the total number of trajectories, length of STN and improvement rate of postoperative movement disorders ( P>0.05). Conclusion:General anesthesia does not affect the MER during STN DBS in the patients with primary PD.

Objective: To investigate the current status of the occupational hazards of welding fume, manganese, and manganese compounds in the welding environment of a large container manufacturing enterprise, as well as the status of occupational health examination of workers, and to provide a basis for improving the welding environment of this enterprise. Methods: In August 2016, July 2017, and August 2018, convenience sampling was used to perform an on-site occupational hygiene survey of the welding workshop for three consecutive years, and welding fume, manganese and, manganese compounds (counted as manganese dioxide) were measured for their workplace exposure concentrations and exposure levels in workers. A comprehensive analysis was performed for the results of occupational health examination. Results: Welding fume, manganese, and manganese compounds in the welding environment gradually increased from 2016 to 2018 (χ²_trend=5.14 and 5.54, P<0.05). The maximum over-standard rate of concentration-short term exposure limit was 43.3% (13/30) for welding fume and 40.0% (12/30) for manganese and its compounds, and the maximum over-standard rate of time-weighted average concentration was 26.7% (8/30) for welding fume and 23.3% (7/30) for manganese and its compounds. Abnormalities were observed in the occupational health examination of welding workers in 2016-2018, among which respiratory system abnormalities (cough, expectoration, and wheezing), nervous system abnormalities (dizziness, fatigue, sleep disorders, amnesia, hyperhidrosis, and palpitations), and electrocardiogram abnormalities (bundle conduction block) had an incidence rate of above 10.0%, and the incidence rate of abnormalities on posterior-anterior X-ray high-kV chest radiograph was close to 8.9% (30/336) . Conclusion There is severe exposure to welding fume, manganese, and manganese compounds among workers in this enterprise, which cause great hazards to the health of workers. It is necessary to strengthen occupational health management, take measures to improve the welding environment, and enhance occupational disease prevention.

Objective: To put forward the suggestion of the occupational contact limit of tributyl phosphate in the air of the workplace. Methods: Data of production and usage, workers' basic information, occupational history, and physical examinations were collected, and the environmental and individual levels of exposure were monitored using fixed-point and individual sampling. The results of the questionnaire and health examination were statistically analyzed using exact probability method of Fisher in the workers exposed to tributyl phosphate and the control group. Results: The results showed that tributyl phosphate was widely distributed in the workplace of production and using enterprises, and the concentration of tributyl phosphate in packaging area was highest at 2.47 mg/m³, and in feeder nose was highest at 2.13 mg/m³. The discomfort symptoms were classified and results showed that tributyl phosphate exposure group of 136 people, all symptoms of 128 people, accounting for 94.44% of the total, the remaining 5.56% of the staff report had psychiatric symptoms or lethargy and irritability skin itching, the control group had no symptoms. There is or not discomfort symptoms in the tributyl phosphate exposure group and the control group was compared with the exact probability of Fisher, and the difference was statistically significant (P<0.05) . The results of healthy physical examination of workers exposed to tributyl phosphate and control group were statistically analyzed by the exact probability method of Fisher. The results showed that there was no significant difference in the results of routine internal medical examination, nervous system examination, skin examination, five senses examination, blood routine, urine routine, lung ventilation, and X ray chest fluoroscopy between the tributyl phosphate exposure group and the control group (P<0.05) . Conclusion The workplace permissible time-weighted tributyl phosphate and short-term exposure limit concentrations in China were set at 2.5 mg/m³ and 5 mg/m³, respectively.

AIM To develop a UPLC-MS/MS method for the simultaneous content determination of adenosine,salidroside,chlorogenic acid,calycosin-7-O-β-D-glycoside,specnuezhenide,calycosin,formononetin,quercetin,apigenin and astragaloside Ⅳ in Zhenqi Fuzheng Capsules (Ligustri lucidi Fructus and Astragali Radix).METHODS The analysis of methanol extract of this drug was performed on a 40 ℃ thermostatic Inertsutain C18 column (75 mm × 3.0 mm,2 μm),with the mobile phase comprising of methanol-acetonitrile-4 mmol/L ammonium acetate flowing at 0.5 mL/min in a gradient elution manner.RESULTS Ten constituents showed good linear relationships within their own ranges (r≥0.996 0),whose average recoveries were 95.1％-104.3％ with the RSDs of less than 4.20％.CONCLUSION This sensitive and specific method can be used for the rapid quality control of Zhenqi Fuzheng Capsules.

60 curved root canals of permanent teeth with angles of curvature ranging from 15°to 40°(Schneider's methodology)were prepared using the instruments of Hyflex CM(HC) and ProTaper(PT) Universal respectively(n=30).Using standardized pre-and post-instrumentation paralleling periapical radiographs,canal curvature was determined by image analysis software and the clinical shaping effect of Hyflex CM and ProTaper rotary NiTi files were compared.The canal curvature in group HC and PT decreased by 4.54°±3.25° and 5.63°±3.84° respectively(between pre-and post-treatment in both groups,P<0.05;between groups,P>0.05).Hyflex CM can meet the clinical necessity for the instrumentation of curved root canals.

Objective To evaluate the value of lung ultrasound score (LUS) in the quantitative assessment of the severity of neonatal respiratory distress syndrome (NRDS) and the value of clinical diagnosis and treatment.Methods Totally 74 NRDS cases and 30 normal neonates were studied.LUS was compared with X-ray examination,clinical data,ventilator assisted ventilation and ventilator parameters.ROC curve was used to calculate sensitivity and specificity of LUS to predict the severity of NRDS and application of invasive ventilator treatment.Results The main findings of the lung ultrasound in NRDS included diffuse distribution of dense B line,disappeared A line,pleural line abnormalities,decreased pulmonary slip sign and pulmonary consolidation.LUS in patients with NRDS was significantly correlated with X ray grades,clinical grades,assisted ventilation mode grades,number of days on ventilator and ventilator parameters (all P＜0.05).LUS value to predict mild,moderate and severe NRDS were 13.0,22.5,and 29.5,respectively.The best cutoff point for LUS to predict the adoptation of invasive assisted ventilation was 22.5,which had sensitivity of 86.0 ％ and specificity of 64.5 ％.Conclusion LUS can be used to diagnose and evaluate the severity of the desease,and to guide the clinical diagnosis and treatment.