Objective To investigate an automated pretreatment technology for detecting levels of free silica in workplace dust. Methods An fully automated pretreatment workstation for detecting free silica levels in workplace dust was developed by integrating graphite-controlled digestion temperature, online-controlled dilution of digestion solutions, and filtration endpoint recognition based on monitoring technology, combined with multi-channel synchronous measurements. Results The fully automatic pretreatment workstation was used to digest and filter 14 standard samples of free silica produced by three institutions, and then detected by pyrophosphate method. The result range of high-, medium-, and low-level free silica standard samples detection was 66.5%-84.8%, 40.0%-44.5%, and 2.1%-24.8%, respectively. The mean relative standard deviations were 3.9%, 1.4% and 1.5%. Conclusion The fully automated pretreatment workstation produced results that met relevant requirements. It can effectively replace the manual digestion and filtration steps of the pyrophosphate method to measure free silica levels in workplace dust and enable rapid detection of free silica in dust samples.

Objective:To explore the clinical efficacy of periosteal induction technique combined with transfer of sural neurovascular flap in treatment of post-traumatic osteomyelitis of calcaneus with soft tissue defect.Methods:Clinical data, from January 2017 to December 2022, of 17 patients in the Army Institute for Traumatic Orthopaedics, the 920th Hospital of Joint Service Force of the Chinese People’s Liberation Amy with post-traumatic calcaneal osteomyelitis combined with soft tissue defect were retrospectively studied. The patients were 11 males and 6 females, with 46.5 (17-68) years in average. All patients received surgical treatment with periosteal induction technique in 2 phased surgies. Thorough debridement, antibiotics blended bone cement filling and wound coverage with sural neurovascular flap were carried out in phase-I surgery; The phase-II surgery were performed at 6-8 weeks after infection control to remove bone cement and then to transfer bone grafts for periosteal induction. After surgery, flap healing and infection control were observed. The infection control, pain improvement, recovery of ankle function and improvement of quality of life were evaluated by comparison of following parameters before and after surgery per phase: infection indicators [white blood cell count (WBC), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP)], Visual Analogue Scale (VAS) score, American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score, and MOS 36-item Short form Health Survey (SF-36, Boston Institute of Health, USA).Results:All 17 patients completed the two-phased surgical treatment, with an average interval of 9.4 (8-16) weeks between phase-I and phase-II surgery. All patients were included in the postoperative follow-up of 25.8 (13-40) months. After debridement in phase-I surgery, the sizes of soft tissue defect were found at 3.0 cm×2.0 cm-6.0 cm×8.0 cm. All flaps survived from the reconstructive surgery of sural neurovascular flap. Postoperative distal flap necroses occurred to 4 patients but all healed after further debridement. Recurrence of postoperative infection occurred to 2 patients and the infection control was achieved after the phase-I rescue surgery. Good outcomes without recurrence of infection were achieved after phase-II surgery. The postoperative follow-up at 1 year after phase-II surgery showed a statistically significant improvement of infection in blood indicators and reductions in VAS score, AOFAS ankle-hindfoot score and SF-36 score in comparison with those before surgery ( P<0.05). In addition to WBC, there were also significant differences in pairwise comparisons between each group at different time points ( P<0.05). Conclusion:In the treatment of post-traumatic calcaneal osteomyelitis with soft tissue defect, a combination of periosteal induction technique and sural neurovascular flap is beneficial to infection control, bone defect reconstruction, recovery of ankle function and improvement of quality of life.

Objective:To explore the clinical efficacy of periosteal induction technique combined with transfer of sural neurovascular flap in treatment of post-traumatic osteomyelitis of calcaneus with soft tissue defect.Methods:Clinical data, from January 2017 to December 2022, of 17 patients in the Army Institute for Traumatic Orthopaedics, the 920th Hospital of Joint Service Force of the Chinese People’s Liberation Amy with post-traumatic calcaneal osteomyelitis combined with soft tissue defect were retrospectively studied. The patients were 11 males and 6 females, with 46.5 (17-68) years in average. All patients received surgical treatment with periosteal induction technique in 2 phased surgies. Thorough debridement, antibiotics blended bone cement filling and wound coverage with sural neurovascular flap were carried out in phase-I surgery; The phase-II surgery were performed at 6-8 weeks after infection control to remove bone cement and then to transfer bone grafts for periosteal induction. After surgery, flap healing and infection control were observed. The infection control, pain improvement, recovery of ankle function and improvement of quality of life were evaluated by comparison of following parameters before and after surgery per phase: infection indicators [white blood cell count (WBC), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP)], Visual Analogue Scale (VAS) score, American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score, and MOS 36-item Short form Health Survey (SF-36, Boston Institute of Health, USA).Results:All 17 patients completed the two-phased surgical treatment, with an average interval of 9.4 (8-16) weeks between phase-I and phase-II surgery. All patients were included in the postoperative follow-up of 25.8 (13-40) months. After debridement in phase-I surgery, the sizes of soft tissue defect were found at 3.0 cm×2.0 cm-6.0 cm×8.0 cm. All flaps survived from the reconstructive surgery of sural neurovascular flap. Postoperative distal flap necroses occurred to 4 patients but all healed after further debridement. Recurrence of postoperative infection occurred to 2 patients and the infection control was achieved after the phase-I rescue surgery. Good outcomes without recurrence of infection were achieved after phase-II surgery. The postoperative follow-up at 1 year after phase-II surgery showed a statistically significant improvement of infection in blood indicators and reductions in VAS score, AOFAS ankle-hindfoot score and SF-36 score in comparison with those before surgery ( P<0.05). In addition to WBC, there were also significant differences in pairwise comparisons between each group at different time points ( P<0.05). Conclusion:In the treatment of post-traumatic calcaneal osteomyelitis with soft tissue defect, a combination of periosteal induction technique and sural neurovascular flap is beneficial to infection control, bone defect reconstruction, recovery of ankle function and improvement of quality of life.

Objective To explore the current research status and frontier hotspot of task-state functional magnetic resonance imaging(fMRI)based on language task in the field of neuropsychiatric diseases. Methods Literature about the application of task-state fMRI based on language task in neuropsychiatric disorders was re-trieved from CNKI,VIP and Web of Science Core Collection databases up to November 12nd,2023.The includ-ed literature was analyzed using CiteSpace and VOSviewer. Results A total of 91 literatures were included.The researches about application of task-state fMRI based on language task in the field of neuropsychiatric disorders were reported since 2002,and developed uniformly;particularly for the diagnosis and treatment of stroke,brain tumors,epilepsy,schizophrenia and other neuropsychiatric disorders,as well as in studies of brain functional connectivity.Research hotspots included mechanisms of aphasia recov-ery,brain functional changes,functional connectivity and language laterality.Journals of psychology,education and sociology played the core role,disciplines such as neurology also contributed to the advancement of this tech-nology. Conclusion Task-state fMRI based on language task demonstrates significant potential in elucidating the mechanisms of how the brain processes language in neuropsychiatric disorders.

ObjectiveTo investigate the infestation and disposal of bedbugs in Shanghai， and provide scientific evidence for comprehensive prevention and control of bedbugs. MethodsA questionnaire survey was conducted in the pest control operations （PCOs） of Shanghai Association for Health Promotion to investigate the infestation and disposal of bedbugs. The questionnaire included basic information of the unit， bedbug disposal experience， type of environment of bedbug infestation， main disposal site， main treatment method， main drug and formulation， and evaluation of the disposal. Chi-square test was used for comparison between groups. ResultsOf the 116 PCOs， 78 （67.24%） had conducted bedbug disposal， and 29.49% had conducted the disposal no less than 5 times in the past three years. The main types of environment of bedbug infestation were dormitories （46.96%）， hotels （18.78%）， and households （20.44%）. Additionally， bedbug infestation was found in nursing homes， prisons and detention centers， and transport vehicles. Bed frame/bed board/mattress （32.42%） was the main site of bedbug infestation， followed by mat （19.63%）. Chemical spraying was the most common method for bedbug disposal （98.72%）， followed by high-temperature steam （11.54%）. The main drug used in chemical spraying were pyrethroids （69.23%）， nicotinoids （60.26%） and organophosphorus （42.31%）， for which the main formulation were suspended insecticides （55.13%）， followed by wettable powders （43.59%） and emulsifiers （43.59%）. Furthermore， 89.74% of the PCOs performed the assessment after disposal， in which the control effect was generally satisfactory. ConclusionBedbug infestation is relatively common in Shanghai. Dormitories， hotels and households are places where bedbug infestation is more likely to occur. Bed frame， bed board and mattress are the most vulnerable sites to bedbug infestation. Combination of physical and chemical methods should be used for disposal of bedbug infestation.

Objective To construct microscale rectangular hydrogel grooves and to investigate the morphology and alignment of human umbilical vein endothelial cells(HUVECs)under spatial constraints.Vascular endothelial cell morphology and alignment are important factors in vascular development and the maintenance of homeostasis.Methods A 4-arm polyethylene glycol-acrylate(PEG-acrylate)hydrogel was used to fabricate rectangular microgrooves of the widths of 60 μm,100 μm,and 140 μm.The sizes and the fibronectin(FN)adhesion of these hydrogel microgrooves were measured.HUVECs were seeded onto the FN-coated microgrooves,while the flat surface without micropatterns was used as the control.After 48 hours of incubation,the morphology and orientation of the cells were examined.The cytoskeleton was labelled with phalloidine and the orientation of the cytoskeleton in the hydrogel microgrooves was observed by laser confocal microscopy.Results The hydrogel microgrooves constructed exhibited uniform and well-defined morphology,a complete structure,and clear edges,with the width deviation being less than 3.5%.The depth differences between the hydrogel microgrooves of different widths were small and the FN adhesion is uniform,providing a micro-patterned growth interface for cells.In the control group,the cells were arranged haphazardly in random orientations and the cell orientation angle was(46.9±1.8)°.In contrast,the cell orientation angle in the hydrogel microgrooves was significantly reduced(P<0.001).However,the cell orientation angles increased with the increase in hydrogel microgroove width.For the 60 μm,100 μm,and 140 μm hydrogel microgrooves,the cell orientation angles were(16.4±2.8)°,(24.5±3.2)°,and(30.3±3.5)°,respectively.Compared to that of the control group(35.7%),the number of cells with orientation angles<30° increased significantly in the hydrogel microgrooves of different widths(P<0.001).However,as the width of the hydrogel microgrooves increased,the number of cells with orientation angles<30° gradually decreased(79.9%,62.3%,54.7%,respectively),while the number of cells with orientation angles between 60°-90° increased(P<0.001).The cell bodies in the microgrooves were smaller and more rounded in shape.The cells were aligned along the direction of the microgrooves and corresponding changes occurred in the arrangement of the cell cytoskeleton.In the control group,cytoskeletal filaments were aligned in random directions,presenting an orientation angle of(45.5±3.7)°.Cytoskeletal filaments were distributed evenly within various orientation angles.However,in the 60 μm,100 μm,and 140 μm hydrogel microgrooves,the orientation angles of the cytoskeletal filaments were significantly decreased,measuring(14.4±3.1)°,(24.7±3.5)°,and(31.9±3.3)°,respectively.The number of cytoskeletal filaments with orientation angles<30° significantly increased in hydrogel microgrooves of different widths(P<0.001).However,as the width of the hydrogel microgrooves increased,the number of cytoskeletal filaments with orientation angles<30° gradually decreased,while the number of cytoskeletal filaments with orientation angles between 60°-90° gradually increased(P<0.001).Conclusion Hydrogel microgrooves can regulate the morphology and orientation of HUVECs and mimic to a certain extent the in vivo microenvironment of vascular endothelial cells,providing an experimental model that bears better resemblance to human physiology for the study of the unique physiological functions of vascular endothelial cells.Nonetheless,the molecular mechanism of spatial constraints on the morphology and the assembly of vascular endothelial cell needs to be further investigated.

Flagella are the main motility structure of Clostridioides difficile that affects the adhesion, colonization, and virulence of C. difficile in the human gastrointestinal tract. The FliL protein is a single transmembrane protein bound to the flagellar matrix. This study aimed to investigate the effect of the FliL encoding gene flagellar basal body-associated FliL family protein (fliL) on the phenotype of C. difficile. The fliL gene deletion mutant (ΔfliL) and its corresponding complementary strains (: : fliL) were constructed using allele-coupled exchange (ACE) and the standard molecular clone method. The differences in physiological properties such as growth profile, antibiotic sensitivity, pH resistance, motility, and spore production ability between the mutant and wild-type strains (CD630) were investigated. The ΔfliL mutant and the : : fliL complementary strain were successfully constructed. After comparing the phenotypes of strains CD630, ΔfliL, and : : fliL, the results showed that the growth rate and maximum biomass of ΔfliL mutant decreased than that of CD630. The ΔfliL mutant showed increased sensitivity to amoxicillin, ampicillin, and norfloxacin. Its sensitivity to kanamycin and tetracycline antibiotics decreased, and the antibiotic sensitivity partially returned to the level of CD630 strain in the : : fliL strain. Moreover, the motility was significantly reduced in the ΔfliL mutant. Interestingly, the motility of the : : fliL strain significantly increased even when compared to that of the CD630 strain. Furthermore, the pH tolerance of the ΔfliL mutant significantly increased or decreased at pH 5 or 9, respectively. Finally, the sporulation ability of ΔfliL mutant reduced considerably compared to the CD630 strain and recovered in the : : fliL strain. We conclude that the deletion of the fliL gene significantly reduced the swimming motility of C. difficile, suggesting that the fliL gene is essential for the motility of C. difficile. The fliL gene deletion significantly reduced spore production, cell growth rate, tolerance to different antibiotics, acidity, and alkalinity environments of C. difficile. These physiological characteristics are closely related to the survival advantage in the host intestine, which is correlated with its pathogenicity. Thus, we suggested that the function of the fliL gene is closely related to its motility, colonization, environmental tolerance, and spore production ability, which consequently affects the pathogenicity of C. difficile.
Humans ; Clostridioides/metabolism* ; Clostridioides difficile/metabolism* ; Bacterial Proteins/metabolism* ; Virulence ; Anti-Bacterial Agents/metabolism*

Organoids are three-dimensional structures formed by self-organizing growth of cells in vitro, which own many structures and functions similar with those of corresponding in vivo organs. Although the organoid culture technologies are rapidly developed and the original cells are abundant, the organoid cultured by current technologies are rather different with the real organs, which limits their application. The major challenges of organoid cultures are the immature tissue structure and restricted growth, both of which are caused by poor functional vasculature. Therefore, how to develop the vascularization of organoids has become an urgent problem. We presently reviewed the progresses on the original cells of organoids and the current methods to develop organoids vascularization, which provide clues to solve the above-mentioned problems.
Humans ; Organoids ; Neovascularization, Pathologic ; Technology

Objective To study the relationship between plasma neurodegenerative protein level and non-motor symptoms(NMS)in PD patients.Methods Eighty-four PD patients served as a PD group and 54age-matched persons undergoing physical examination served as a control group. The NMS of PD patients were assessed according to the HAMD scale.The plasma levels of tau,p-tau181,Aβ-42andα-syn were measured by ELISA and analyzed by Spearman correlation analysis and binary logistic regression analysis respectively.Results The FSS score and plasmaα-syn level were significantly higher while the plasma Aβ-42level was significantly lower in PD group than in control group(3.22±1.68 vs 1.89±1.16,P=0.000;320.00±64.91ng/L vs 277.78±52.75ng/L,P=0.000;267.61±77.75ng/L vs 321.80±49.41ng/L,P=0.001).No significant difference was detected in plasma tau and p-tau181levels between the two groups(P＞0.05).The plasmaα-syn level was positively related with the FSS score(r=0.237,P=0.030)and was an influencing factor of FSS(OR=1.019,95％CI:1.006-1.032,P=0.004).Conclusion Plasma neurodegenerative protein level is related with NMS and plasmaα-syn level is a peripheral biomarker for fatigue in PD patients.

Objective To detect the expression of costimulatory molecules B7-H4 and programmed death ligand 1 (PD-L1) in bladder cancer,and to explore the correlation between them and clinicopathological features of bladder cancer.Methods Immunohistochemical staining was used to detect the expression of B7-H4 and PD-L1 in 98 cases of bladder urothelial carcinoma,which were confirmed by pathology from August 2014 to December 2015 in our hospital.There were 23 females,aged 45-82 years,with an average age of 67.8 years.Among them,42 cases of adjacent normal tissues were used as controls.The clinical stage,histological grade and recurrence of bladder cancer were collected,and the correlation between them was analyzed.Results The positive rates of B7-H4 and PD-L1 in bladder urothelial carcinoma were 54.1％ (53/98) and 59.2％ (58/98),respectively,and there was no expression in normal bladder tissues (P ＜ 0.05).The positive expression rate of B7-H4 in muscle invasive bladder cancer (MIBC) patients was higher than that in non-muscle invasive bladder cancer (NMIBC) [73.5％ (25/34) vs.43.8％ (28/64),P =0.005].The positive expression rate of B7-H4 in high-grade patients was higher than that of low-grade [70.0％ (21/30) vs.47.1％ (32/68),P =0.036].The expression rate of B7-H4 in high-risk group was higher than that of low-intermediate risk group [57.1％ (20/35) vs.27.6％ (8/29),P =0.018].The positive expression rate of PD-L1 in patients with MIBC was higher than that in NMIBC [79.4％ (27/34) vs.48.4％ (31/64),P =0.003].The PD-L1 expression rate of histological high-level group was higher than that of low-level group [73.3％ (22/30) vs.52.9％ (36/68)],but the difference was not statistically significant (P =0.058).The PD-L1 expression rate in high-risk group was 68.6％ (24/35),and also higher than low-middle group 24.1％ (7/29) (P ＜ 0.05).There was a positive correlation between the expression of B7-H4 and PD-L1 in bladder urothelial carcinoma (r =0.318,P=0.002).The combined recurrence rate of the two groups was significantly higher than that of the negative expression of the two groups [66.7％ (14/21) vs.30.8％ (8/26),P=0.014].Conclusions The expression of B7-H4 and PD-L1 is up-regulated in bladder urothelial carcinoma,which is closely related to the clinical stage,histological grade,risk classification and recurrence of NMIBC.