The efficient production of L-glutamate is dependent on the product's rapid efflux, hence researchers have recently concentrated on artificially modifying its transport system and cell membrane wall structure. Considering the unique composition and structure of the cell wall of Corynebacterium glutamicum, we investigated the effects of CmpLs on L-glutamate synthesis and transport in SCgGC7, a constitutive L-glutamate efflux strain. First, the knockout strains of CmpLs were constructed, and it was confirmed that the deletion of CmpL1 and CmpL4 significantly improved the performance of L-glutamate producers. Next, temperature-sensitive L-glutamate fermentation with the CmpL1 and CmpL4 knockout strains were carried out in 5 L bioreactors, where the knockout strains showcased temperature-sensitive characteristics and enhanced capacities for L-glutamate production under high temperatures. Notably, the CmpL1 knockout strain outperformed the control strain in terms of L-glutamate production, showing production and yield increases of 69.2% and 55.3%, respectively. Finally, the intracellular and extracellular metabolites collected at the end of the fermentation process were analyzed. The modification of CmpLs greatly improved the L-glutamate excretion and metabolic flux for both L-glutamate production and transport. Additionally, the CmpL1 knockout strain showed decreased accumulation of downstream metabolites of L-glutamate and intermediate metabolites of tricarboxylic acid (TCA) cycle, which were consistent with its high L-glutamate biosynthesis capacity. In addition to offering an ideal target for improving the stability and performance of the industrial strains for L-glutamate production, the functional complementarity and redundancy of CmpLs provide a novel target and method for improving the transport of other metabolites by modification of the cell membrane and cell wall structures in C. glutamicum.
Corynebacterium glutamicum/genetics* ; Glutamic Acid/biosynthesis* ; Fermentation ; Metabolic Engineering ; Bacterial Proteins/metabolism* ; Bioreactors/microbiology* ; Gene Knockout Techniques

Objective To investigate the causes for the continuous detection of Pseudomonas fluorescens(P.fluo-rescens)from bronchoalveolar lavage fluid(BALF)in pediatric department of a hospital,formulate intervention measures and evaluate its effectiveness,and provide basis for improving the whole process infection control of fiber bronchoscopy.Methods Epidemiological investigation was conducted on three children from whose BALF P.fluorescens were detected in May 3-6,2024.The comprehensive methods were adopted,including case revie-wing,on-site process tracking,environmental hygiene monitoring,laboratory testing on disinfectant sterilization effect,fiber bronchoscope structure maintenance and checking,etc.Risks were identified and targeted interventions were implemented.Results Among the 5 pediatric patients who underwent fiber bronchoscopy within 4 days,P.fluorescens was detected from BALF of 3 cases,with a detection rate of 60.0％.The children were 5-8 years old and were admitted to the hospital due to lobar pneumonia.They underwent fiber bronchoscopy from the day of admission to the second day,and bacterial strains were clinically determined to be contaminated strains.Environ-mental sampling showed that the detection rate of P.fluorescens at sampling points such as fiber bronchoscope and enzyme solution storage tank was 15.7％(8/51).After implementing intervention,no target bacteria were detected again,and the difference was statistically significant(P＜0.05).From January 1 to May 2,2024,71 BALF from pediatric department were not detected P.fluorescens;From May 3 to 6,among 5 detected BALF,3 were detected P.fluorescens;After intervention(May 16 to December 31),no specimen was detected P.fluorescens.Conclusion This event is a pseudo-outbreak caused by fiber bronchoscope damage as well as improper cleaning and disinfection procedures.Through collaborative investigation and timely intervention by multiple departments,the event was ef-fectively controlled.

Objective To investigate the causes for the continuous detection of Pseudomonas fluorescens(P.fluo-rescens)from bronchoalveolar lavage fluid(BALF)in pediatric department of a hospital,formulate intervention measures and evaluate its effectiveness,and provide basis for improving the whole process infection control of fiber bronchoscopy.Methods Epidemiological investigation was conducted on three children from whose BALF P.fluorescens were detected in May 3-6,2024.The comprehensive methods were adopted,including case revie-wing,on-site process tracking,environmental hygiene monitoring,laboratory testing on disinfectant sterilization effect,fiber bronchoscope structure maintenance and checking,etc.Risks were identified and targeted interventions were implemented.Results Among the 5 pediatric patients who underwent fiber bronchoscopy within 4 days,P.fluorescens was detected from BALF of 3 cases,with a detection rate of 60.0％.The children were 5-8 years old and were admitted to the hospital due to lobar pneumonia.They underwent fiber bronchoscopy from the day of admission to the second day,and bacterial strains were clinically determined to be contaminated strains.Environ-mental sampling showed that the detection rate of P.fluorescens at sampling points such as fiber bronchoscope and enzyme solution storage tank was 15.7％(8/51).After implementing intervention,no target bacteria were detected again,and the difference was statistically significant(P＜0.05).From January 1 to May 2,2024,71 BALF from pediatric department were not detected P.fluorescens;From May 3 to 6,among 5 detected BALF,3 were detected P.fluorescens;After intervention(May 16 to December 31),no specimen was detected P.fluorescens.Conclusion This event is a pseudo-outbreak caused by fiber bronchoscope damage as well as improper cleaning and disinfection procedures.Through collaborative investigation and timely intervention by multiple departments,the event was ef-fectively controlled.

Objective To investigate the serum levels of homocysteine(HCY)and cystatin C(Cys-C)in patients with antineutrophil cytoplasmic antibody(ANCA)-associated vasculitis(AAV)associated renal injury,and to explore the diagnostic value of combined detection of HCY and Cys-C in ANCA-associated renal injury.Methods We selected 70 ANCA-positive patients with renal injury admitted to the Department of Nephrology in our hospital from January 2016 to January 2019.Another 48 patients with ANCA-positive but not complicated with renal injury were in simple ANCA positive group while 50 patients with physical examination who came to the hospital during the same period were in control group.Healthy patients served as the control group.By comparing the differences in the test results of different groups,the area under the curve(AUC)of each test index for ANCA-positive renal injury was determined,and the correlation and diagnostic value of combined detection of HCY and Cys-C for ANCA-related renal injury were calculated.Results The levels of HCY and Cys-C in the ANCA positive combined with renal injury group,the pure ANCA positive group,and the control group differed significantly(P＜0.05).The positive rates of HCY and Cys-C in the two groups were significantly different(P＜0.05).The sensitivity and specificity of HCY and Cys-C in combination for the diagnosis of ANCA-positive related renal injury were significantly higher than those of the above indicators alone(P＜0.05).ROC curve results showed that HCY combined with Cys-C had a better diagnosis for patients with positive ANCA and associated renal injury and a higher correlation.Conclusion HCY combined with Cys-C is closely correlated to ANCA-related renal injury,and the combined detection has a positive effect on the diagnosis of patients with ANCA-positive related renal injury,and has guiding value for the treatment of patients.

With increasing age, more and more patients with posterior chamber intraocular lens (ICL) implantation are facing the threat of cataracts to their visual acuity.When examining the eyes of cataract patients after ICL surgery, attention should be paid to whether the density of corneal endothelial cells is greater than 2 000 cells/mm 2, the state of the anterior chamber angle, and whether there are fundus abnormalities such as retinal detachment and choroidal neovascularization.When conducting eye biometry measurement, attention should be paid to the measurement starting and ending lines of anterior chamber depth and lens thickness.If patients undergo ICL combined with corneal refractive surgery, they should be examined with two or more devices to obtain corneal refractive power according to the examination requirements after corneal laser vision correction.When selecting the type of intraocular lens, consideration should be given to the histological characteristics of high myopia.Compared to C- and L- loops, plate-haptic is relatively more stable in patients with high myopia accompanied by large capsules and larger diameters of continuous curvilinear capsulorhexis.Kane, Barrett Universal Ⅱ, Olsen, Hill-RBF formulas for calculating the refractive power of intraocular lenses are more accurate in people with long axial length.It is recommended to perform ICL removal simultaneously with phacoemulsification and intraocular lens implantation, preferably with a surgical incision greater than 2.6 mm.Femtosecond laser assisted cataract extraction surgery, although superior to traditional phacoemulsification in reducing corneal endothelial cell loss, reducing corneal edema, and high-quality capsulorhexis, can cause incomplete capsulorhexis and fragmentation due to the cavitation bubbles, manual adjustment of location, and the impact of lower vault.It is recommended to use it with caution.Ophthalmologists should fully understand and pay attention to the characteristics and difficulties of cataract surgery after ICL surgery, communicate fully with patients, and make personalized surgery to achieve better visual outcomes.

ObjectiveTo explore the effects of phillygenin （PHI） on the inflammation in L02 cells induced by lipopolysaccharide （LPS） and adenosine triphosphate （ATP） and the expression of purinergic 2X7 receptor （P2X7R）， NOD-like receptor family pyrin domain containing 3 （NLRP3）， and nuclear factor kappa B （NF-κB） expression. MethodIn this study， the inflammation model was induced in L02 cells by 100 μg·L^-1 LPS treatment for 24 h and 5 mmoL·L^-1 ATP treatment for 5 h. The cells in the PHI groups were cultured with PHI （100， 50， 25 mg·L^-1） for 6 h in the LPS treatment period， followed by LPS treatment for another 18 h. After ATP treatment for 5 h， the mRNA and protein expression of interleukin-1β （IL-1β）， interleukin-18 （IL-18）， P2X7R， NLRP3， Caspase-1 precursor （pro-Caspase-1）， cleaved Caspase-1， NF-κB， and NF-κB inhibitor protein α （IκBα） in L02 cells was detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. Molecular docking was used to predict whether P2X7R could bind to PHI， and DCFH-DA was employed to detect the accumulation of reactive oxygen species （ROS） in cells. P2X7R was silenced by small interfering ribonucleic acid （siRNA）， and then the mRNA expression of IL-1β， IL-18， P2X7R， NLRP3， Caspase-1， NF-κB， and IκBα was detected by Real-time PCR. ResultReal-time PCR and Western blot showed that compared with the normal group， the model group showed increased expression of IL-1β and IL-18 （P<0.05）， and compared with the model group， the PHI groups showed down-regulated IL-1β， IL-18 mRNA and protein expression （P<0.05）. Molecular docking suggested a good binding effect of PHI to P2X7R. Real-time PCR and Western blot analysis showed that the expression of P2X7R in the model group was significantly up-regulated compared with that in the normal group （P<0.05）， and compared with the model group， the PHI groups showed down-regulated mRNA and protein expression of P2X7R （P<0.05）. DCFH-DA results showed that compared with the normal group， the model group showed increased content of ROS （P<0.05）， and compared with the model group， the PHI groups decreased the accumulation of ROS （P<0.05）. As demonstrated by Real-time PCR and Western blot， compared with the normal group， the model group showed increased expression of NLRP3 inflammasome and NF-κB （P<0.05）， and compared with the model group， the PHI groups significantly decreased the mRNA and protein expression of NLRP3 and cleaved Caspase-1， and up-regulated the mRNA and protein expression of NF-κB and IκBα （P<0.05）. Real-time PCR analysis showed that compared with the results in the model group， after silencing P2X7R by siRNA， the mRNA expression of IL-1β， IL-18， P2X7R， NLRP3， Caspase-1， NF-κB， and IκBα was decreased （P<0.05）. PHI exerted the same effect， and the mRNA expression was further reduced after the combination of them. ConclusionPHI is presumed to suppress the expression of the NLRP3/NF-κB signaling pathway by down-regulating upstream P2X7R to alleviate the LPS/ATP-induced inflammation in L02 cells， suggesting that P2X7R may be the target of PHI against inflammation.

Objective: To observe the clinical efficacy of electroacupuncture (EA) in controlling myopia in children and its effect on retinal blood flow. Methods: Sixty-eight myopic children were randomly divided into an observation group and a control group, with 34 cases in each group. The control group was given auricular acupressure treatment alone, and the observation group was treated with EA once a week in addition to the treatment used in the control group. The spherical equivalent refraction (SER) and axial length (AL) were measured at baseline, and after 3 months and 6 months of treatment. Optical coherence tomography angiography was used to measure the vessel density (VD) and perfusion density (PD) in the surface layer of the retina. Results: After 3 months and 6 months of treatment, the changes in SER between the two groups were not statistically significant (P>0.05). After 3 months of treatment, the changes in AL between the two groups were not statistically significant (P>0.05); after 6 months of treatment, the change amount of AL in the observation group was smaller than that in the control group (P<0.05); after 3 months and 6 months of treatment, the changes in VD and PD in the surface layer of the retina in the observation group were significantly greater than those in the control group (P<0.01). Conclusion: EA treatment once a week for 6 months can delay the increase of AL and improve the retinal surface blood flow in myopic children.

In the study of oral orthodontics, the dental tissue models play an important role in finite element analysis results. Currently, the commonly used alveolar bone models mainly have two kinds: the uniform and the non-uniform models. The material of the uniform model was defined with the whole alveolar bone, and each mesh element has a uniform mechanical property. While the material of the elements in non-uniform model was differently determined by the Hounsfield unit (HU) value of computed tomography (CT) images where the element was located. To investigate the effects of different alveolar bone models on the biomechanical responses of periodontal ligament (PDL), a clinical patient was chosen as the research object, his mandibular canine, PDL and two kinds of alveolar bone models were constructed, and intrusive force of 1 N and moment of 2 Nmm were exerted on the canine along its root direction, respectively, which were used to analyze the hydrostatic stress and the maximal logarithmic principal strain of PDL under different loads. Research results indicated that the mechanical responses of PDL had been affected by alveolar bone models, no matter the canine translation or rotation. Compared to the uniform model, if the alveolar bone was defined as the non-uniform model, the maximal stress and strain of PDL were decreased by 13.13% and 35.57%, respectively, when the canine translation along its root direction; while the maximal stress and strain of PDL were decreased by 19.55% and 35.64%, respectively, when the canine rotation along its root direction. The uniform alveolar bone model will induce orthodontists to choose a smaller orthodontic force. The non-uniform alveolar bone model can better reflect the differences of bone characteristics in the real alveolar bone, and more conducive to obtain accurate analysis results.
Biomechanical Phenomena ; Computer Simulation ; Finite Element Analysis ; Humans ; Models, Biological ; Periodontal Ligament ; Stress, Mechanical ; Tooth Movement Techniques

Mandible is an important bone of the head and neck. Mandibular defects not only affect patient's face, but also impede patient's daily functions, such as chewing, speech, and so on. Fibular transplantation for mandibular reconstruction is the common method, which requests high accuracy of bone positioning and posture adjustment. Therefore, a robotic system for mandibular reconstruction surgery with fibula flaps was designed to assist surgeons to hold and locate bones, and the model comparison experiments were conducted. The results showed that the robotic system can assist surgeons for mandibular reconstruction to improve quality of surgery.
Bone Transplantation ; methods ; standards ; Fibula ; transplantation ; Humans ; Mandible ; surgery ; Mandibular Reconstruction ; methods ; Robotics ; standards ; Surgery, Computer-Assisted

Objective To integrate the best available evidence of preoperative care for ophthalmic patients undergoing day surgery into clinical practice,so as to improve the quality of nursing.Methods This project was performed in an ophthalmic day surgery ward,guided by the Joanna Briggs Institute(JBI)Practice Application of Clinical Evidence System (PACES) program which includes 3 procedures:baseline audit,evidence implementation,and re-audit.Eight audit criteria were made based on evidences from JBI.Twelve nurses and 267 patients were enrolled in baseline audit,and 12 nurses and 234 patients were recruited for re-audit after evidence implementation.Results Six of the audit criteria that nurses showed poor compliance in the baseline audit(0％～28.5％),such as providing preoperative telephone screening and reducing preoperative waiting time through better preparation and planning,were significantly improved in the re-audit(71.43％～100％).The preoperative waiting time was shortened from 109.70±18.34 minutes to 88.22±9.91 minutes;the correct rates of operation-related knowledge in patients increased from 63.0％ to 85.09％;the patients' satisfaction scored 84.23±17.01 in baseline audit and 95.02±7.32 in the re-audit(P＜0.001).Conclusion Application of evidence-based preoperative care of ophthalmic patients undergoing day surgery can improve nurse's behavior and compliance with the implementation of evidence into clinical practice,and increase patient satisfaction.Continuous audits are needed to continuously improve clinical nursing quality.