ObjectiveTo investigate the regulatory effect of overexpressed protein tyrosine phosphatase non-receptor type 2 （Ptpn2） on the inflammatory response of mouse alveolar macrophages （MH-S） induced by SiO₂. MethodsCells with overexpressed Ptpn2 were constructed and induced by SiO₂. The experimental groups were divided into four groups： the negative control group with an empty vector （NC）， the overexpressed Ptpn2 group （P）， the negative control group with an empty vector + SiO₂ induction （NS）， and the overexpressed Ptpn2 + SiO₂ induction group （PS）. Isobaric tags for relative and absolute quantification （iTRAQ） combined with liquid chromatography-tandem mass spectrometry （LC-MS/MS） were used to screen differential proteins， followed by Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） database analyses. Immunofluorescence staining was used to detect the expressions of Tumor necrosis factor （TNF） α， Gasdermin D （GSDMD）， and Transforming growth factor （TGF）-β1. Western blot was used to detect the protein expression levels of PTPN2， Toll-like receptor 4 （TLR4）， tumor necrosis factor-α （TNF-α）， nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3）， and proteins related to the TGF-β1 signaling pathway in the cells of each group. ResultsiTRAQ results identified 144 differential proteins among the four groups. GO analysis showed that in biological processes （BP）， these differential proteins were mainly enriched in IκB kinase/nuclear factor-κB （NF-κB） signaling， cell activation and signal transduction involved in immune responses， and regulation of receptor signaling pathways by signal transducer and activator of transcription （STAT）， etc. KEGG analysis revealed that the differential proteins were mainly enriched in Toll-like receptor signaling pathway， NF-κB signaling pathway， NOD-like receptor signaling pathway， TGF-β signaling pathway， and TNF signaling pathway. The results of immunofluorescence staining showed that compared with the NC group， the expressions of TNF α， GSDMD， and TGF-β1 in the cells of the NS group increased （P < 0.05）； compared to the NS group， the expression of the aforementioned proteins in the PS group decreased in cellular proteins（P < 0.05）. The results of Western blot showed that compared with the NC group， the protein expression levels of PTPN2， p-NF-κB，MyD88，TLR4，NLRP3，GSDMD，Caspase-1，IL-1β， TGF-βR1， TGF-βR，p-Smad2/3 in the NS group were significantly upregulated （P < 0.05）； compared with the NS group， the expression levels of the aforementioned proteins in the PS group were significantly downregulated （P < 0.05）. ConclusionOverexpression of Ptpn2 can inhibit the protein expressions of TLR4-TNF-α signaling， NLRP3 signaling， and TGF-β1 signaling closely related to inflammatory response in SiO₂-mediated MH-S macrophages.

Objective:To observe the effects of peptidylarginine deiminase 2 (PAD2) inhibitor AFM-30a on silicotic mice and its possible mechanisms.Methods:In May 2022, 40 SPF male C57BL/6J mice were randomly divided into control group, AFM-30a group, silicosis model group and AFM-30a treatment group, with 10 mice in each group. Silicosis model group and AFM-30a treatment group were perfused with silicon dioxide (SiO 2) suspension (10 mg/piece, 50 μl), and the other groups were perfused with an equal amount of sodium chloride solution. After 2 weeks, AFM-30a group and AFM-30a treatment group were intraperitoneally injected AFM-30a (20 mg/kg, 100 μl) daily, and mice of other groups were injected with equal amounts of sodium chloride solution for 4 weeks. Mouse RAW264.7 monocytes/macrophages were cultured in vitro and divided into blank control group, AFM-30a group (5 μmol/L), SiO 2 group (200 μg/ml), and SiO 2+AFM-30a group (200 μg/ml SiO 2 induction for 12 h, followed by 5 μmol/L AFM-30a treatment for 12 h). As well as blank control group, vimentin (Vim) group (2 μg/ml), citrullinated vimentin (Cit-Vim) group (2 μg/ml), and Cit-Vim+TLR4-C34 group (10 μmol/L TLR4-C34 treatment for 1 h, followed by 2 μg/ml Cit-Vim induction for 24 h). Hematoxylin Eosin (HE) and Masson staining were used to observe the pathological morphology of lung. The lung fieldclarity and lung texture of each group was observed by micro-CT. The number of positive cells was detected by tartrate resistant acid phosphatase (TRAP) staining. The localization and expression levels of PAD2, Cit-Vim, toll-like receptor 4 (TLR4) signaling and receptor activator of nuclear factor-κB ligand (RANKL) signaling proteins were measured by Immunofluorescence staining and Western blotting in vitro and in vivo. The experimental data were all presented as Mean±SD. A completely random design of one-way analysis of variance was used among the groups. The pduo comparison was performed using LSD test for homogeneity of variance and Tamhane's test for inconsistency. Results:Compared with the control group, the silicosis model group showed the formation of silicon nodules accompanied by collagen deposition, the silicosis model group showed thickened, and several high-density shadows of varying sizes in the lung field, and the number of TRAP positive cells in silicosis model group were increased significantly, the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signal-related proteins were also significantly increased in silicosis groupmodel ( P<0.05). Compared with the silicosis model group, the AFM-30a treatment group reduced deposition of collagen in lung, and the number of TRAP positive cells was decreased in AFM-30a treatment group. The expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins were significantly decreased in AFM-30a treatment group ( P<0.05). In vitro, compared with the blank control group, the number of TRAP positive cells and the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins in the SiO 2 group were significantly increased ( P<0.05). Compared with the SiO 2 group, the number of TRAP positive cells and the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins in the SiO 2+AFM-30a group were significantly decreased ( P<0.05). Compared with the blank control group, the expression levels of TLR4 and RANKL signaling related proteins in the Cit-Vim group were significantly increased ( P<0.05). Compared with the Cit-Vim group, the expression levels of TLR4 and RANKL signaling related proteins in the Cit-Vim+TLR4-C34 group were significantly decreased ( P<0.05) . Conclusion:PAD2 inhibitor AFM-30a may play an antagonisticrole in silicotic fibrosis in mice by potentialregulating TLR4 and RANKL signaling pathways.

Objective:To observe the effects of peptidylarginine deiminase 2 (PAD2) inhibitor AFM-30a on silicotic mice and its possible mechanisms.Methods:In May 2022, 40 SPF male C57BL/6J mice were randomly divided into control group, AFM-30a group, silicosis model group and AFM-30a treatment group, with 10 mice in each group. Silicosis model group and AFM-30a treatment group were perfused with silicon dioxide (SiO 2) suspension (10 mg/piece, 50 μl), and the other groups were perfused with an equal amount of sodium chloride solution. After 2 weeks, AFM-30a group and AFM-30a treatment group were intraperitoneally injected AFM-30a (20 mg/kg, 100 μl) daily, and mice of other groups were injected with equal amounts of sodium chloride solution for 4 weeks. Mouse RAW264.7 monocytes/macrophages were cultured in vitro and divided into blank control group, AFM-30a group (5 μmol/L), SiO 2 group (200 μg/ml), and SiO 2+AFM-30a group (200 μg/ml SiO 2 induction for 12 h, followed by 5 μmol/L AFM-30a treatment for 12 h). As well as blank control group, vimentin (Vim) group (2 μg/ml), citrullinated vimentin (Cit-Vim) group (2 μg/ml), and Cit-Vim+TLR4-C34 group (10 μmol/L TLR4-C34 treatment for 1 h, followed by 2 μg/ml Cit-Vim induction for 24 h). Hematoxylin Eosin (HE) and Masson staining were used to observe the pathological morphology of lung. The lung fieldclarity and lung texture of each group was observed by micro-CT. The number of positive cells was detected by tartrate resistant acid phosphatase (TRAP) staining. The localization and expression levels of PAD2, Cit-Vim, toll-like receptor 4 (TLR4) signaling and receptor activator of nuclear factor-κB ligand (RANKL) signaling proteins were measured by Immunofluorescence staining and Western blotting in vitro and in vivo. The experimental data were all presented as Mean±SD. A completely random design of one-way analysis of variance was used among the groups. The pduo comparison was performed using LSD test for homogeneity of variance and Tamhane's test for inconsistency. Results:Compared with the control group, the silicosis model group showed the formation of silicon nodules accompanied by collagen deposition, the silicosis model group showed thickened, and several high-density shadows of varying sizes in the lung field, and the number of TRAP positive cells in silicosis model group were increased significantly, the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signal-related proteins were also significantly increased in silicosis groupmodel ( P<0.05). Compared with the silicosis model group, the AFM-30a treatment group reduced deposition of collagen in lung, and the number of TRAP positive cells was decreased in AFM-30a treatment group. The expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins were significantly decreased in AFM-30a treatment group ( P<0.05). In vitro, compared with the blank control group, the number of TRAP positive cells and the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins in the SiO 2 group were significantly increased ( P<0.05). Compared with the SiO 2 group, the number of TRAP positive cells and the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins in the SiO 2+AFM-30a group were significantly decreased ( P<0.05). Compared with the blank control group, the expression levels of TLR4 and RANKL signaling related proteins in the Cit-Vim group were significantly increased ( P<0.05). Compared with the Cit-Vim group, the expression levels of TLR4 and RANKL signaling related proteins in the Cit-Vim+TLR4-C34 group were significantly decreased ( P<0.05) . Conclusion:PAD2 inhibitor AFM-30a may play an antagonisticrole in silicotic fibrosis in mice by potentialregulating TLR4 and RANKL signaling pathways.

The COVID-19 epidemic has spread to the whole world for three years and has had a serious impact on human life, health and economic activities. China's epidemic prevention and control has gone through the following stages: emergency unconventional stage, emergency normalization stage, and the transitional stage from the emergency normalization to the "Category B infectious disease treated as Category B" normalization, and achieved a major and decisive victory. The designated hospitals for prevention and control of COVID-19 epidemic in Tianjin has successfully completed its tasks in all stages of epidemic prevention and control, and has accumulated valuable experience. This article summarizes the experience of constructing a hospital infection prevention and control system during the "Category B infectious disease treated as Category A" period in designated hospital. The experience is summarized as the "Cluster" hospital infection prevention and control system, namely "three rings" outside, middle and inside, "three districts" of green, orange and red, "three things" before, during and after the event, "two-day pre-purification" and "two-director system", and "one zone" management. In emergency situations, we adopt a simplified version of the cluster hospital infection prevention and control system. In emergency situations, a simplified version of the "Cluster" hospital infection prevention and control system can be adopted. This system has the following characteristics: firstly, the system emphasizes the characteristics of "cluster" and the overall management of key measures to avoid any shortcomings. The second, it emphasizes the transformation of infection control concepts to maximize the safety of medical services through infection control. The third, it emphasizes the optimization of the process. The prevention and control measures should be comprehensive and focused, while also preventing excessive use. The measures emphasize the use of the least resources to achieve the best infection control effect. The fourth, it emphasizes the quality control work of infection control, pays attention to the importance of the process, and advocates the concept of "system slimming, process fattening". Fifthly, it emphasizes that the future development depends on artificial intelligence, in order to improve the quality and efficiency of prevention and control to the greatest extent. Sixth, hospitals need to strengthen continuous training and retraining. We utilize diverse training methods, including artificial intelligence, to ensure that infection control policies and procedures are simple. We have established an evaluation and feedback mechanism to ensure that medical personnel are in an emergency state at all times.

Background The senescence of alveolar type II epithelial cells is an important driving factor for the progression of silicotic fibrosis, and the regulatory effects of oxamate on the senescence of alveolar type II epithelial cells is still unclear. Objective To explore whether lactate dehydrogenase inhibitor oxamate can alleviate silicotic fibrosis in mice by inhibiting senescence of alveolar type II epithelial cellsMethods This study was divided into two parts: in vivo experiments and in vitro experiments. In the first part, forty SPF C57BL/6J male mice were randomly divided into four groups with 10 in each group: control group, silicosis model group, low-dose oxamate treatment group, and high-dose oxamate treatment group. The silicotic mouse model was established by intratracheal instillation of 50 μL SiO₂ suspension (100 mg·mL⁻¹). The treatment models were prepared by intraperitoneal injection of 100 μL oxamate (225 mmol·L⁻¹ and 1125 mmol·L⁻¹). In the second part, induction of MLE-12 mouse alveolar type II epithelial cells was conducted with SiO₂. The in vitro experimental groups were ① SiO₂ induction groups: control group, 50 μg·mL⁻¹ SiO₂ group, 100 μg·mL⁻¹ SiO₂ group, and 200 μg·mL⁻¹ SiO₂ group, and ② oxamate treatment groups: control group, SiO₂ group (100 μg·mL⁻¹), low-dose oxamate (25 mmol·L⁻¹) treatment group, and high-dose oxamate (50 mmol·L⁻¹) treatment group. Pathological morphology of lung tissues was evaluated after hematoxylin-eosin (HE) staining; deposition of collagen in lung tissues was evaluated after sirius red staining; positive co-expression of prosurfactant protein C (Pro-SPC) and β-galactosidase was detected by immunofluorescence staining; positive expression of β-galactosidase in MLE-12 cells was detected by immunofluorescence staining. The protein expression levels of collagen type I (CoL I), fibronectin1 (FN1), hexokinase 2 (HK2), pyruvate kinase isozyme type M2 (PKM2), lactate dehydrogenase A (LDHA), p-ataxia telangiectasia and Rad3-related kinase (ATR), and cyclin-dependent kinase inhibitors p21, and p16 were detected by Western blotting. Results Compared with the control group, the protein expression levels of HK2, PKM2, LDHA, p-ATR, p21, and p16 were significantly upregulated in the silicosis model group and the SiO₂-induced MLE-12 cells (P＜0.05). The in vivo studies showed that, compared with the control group, the silicon nodule area, the collagen deposition area, the proportion of β-galactosidase positive cells, and the protein expression levels of CoL I, FN1, LDHA, p-ATR, p21, and p16 were significantly upregulated in the silicosis model group (P＜0.05). Compared with the silicosis model group, the oxamate treatment groups showed significant downregulation of the silicon nodule area, the collagen deposition area, the proportion of β-galactosidase positive cells, and the the CoL I, FN1, LDHA, p-ATR, p21, and p16 protein expression levels, and the high-dose oxamate treatment group showed a higher efficacy on these indicators than the low-dose oxamate treatment group (P＜0.05). The in vitro studies showed that, compared with the control group, the proportion of β-galactosidase positive cells and the protein expression levels of p-ATR, p21, and p16 were significantly upregulated in the SiO₂-induced group (P＜0.05). Compared with the SiO₂ group, the proportion of β-galactosidase positive cells and the LDHA, p-ATR, p21 and p16 protein expression levels were significantly downregulated in the oxamate treatment groups, and the high-dose oxamate treatment group showed a higher efficacy on these indicators than the low-dose oxamate treatment group (P＜0.05). Conclusion Lactate dehydrogenase inhibitor oxamate can alleviate silicotic fibrosis in mice by inhibiting the senescence of alveolar type II epithelial cells.

Objective: To investigate the status of hair loss and analyze the influencing factors among university students in Hangzhou City, so as to provide insights into the management of hair loss among university students. Methods: University students were recruited using a convenient sampling method from 4 universities in Hangzhou City in June 2021. The basic characteristics and life styles were collected using online questionnaire surveys. Self-reported hair loss was evaluated using the grading scales for loss of hair (Hamilton-Norwood scale for males and modified Ludwig scale for females), and factors affecting self-reported hair loss were identified among university students using the multivariable logistic regression model. Results: A total of 1 060 questionnaires were allocated, and 1 038 valid questionnaires were recovered, with an effective recovery rate of 97.92%. The respondents included 391 males ( 37.67% ) and 647 females ( 62.33% ), and 463 respondents ( 44.61% ) reported hair loss, including 431 students with mild hair loss ( 93.09% ). Multivariable logistic regression analysis showed that university students in their fourth or fifth years ( OR=1.721, 95%CI: 1.126-2.630 ), art specialty ( OR=0.411, 95%CI: 0.207-0.816 ), overweight or obesity (OR=1.685, 95%CI: 1.050-2.704), diet taste ( sweet: OR=2.131, 95%CI: 1.370-3.316; spicy: OR=1.510, 95%CI: 1.028-2.218; greasy: OR=3.023, 95%CI: 2.015-4.537 ), feeling nervous/anxious (occasionally: OR=1.891, 95%CI: 1.087-3.289; frequently: OR=2.487, 95%CI: 1.337-4.626 ), smoking ( occasionally: OR=1.906, 95%CI: 1.067-3.405; frequently: OR=1.983, 95%CI: 1.050-3.746), family history of hair loss ( OR=1.506, 95%CI: 1.075-2.110 ), perming/dyeing hair ( occasionally: OR=1.795, 95%CI: 1.280-2.517; frequently: OR=3.282, 95%CI: 1.736-6.204), self-perceived oily hair/scalp in the past three months (slightly increased: OR=1.980, 95%CI: 1.477-2.653; significantly increased: OR=5.347, 95%CI: 2.956-9.670) were factors affecting self-reported hair loss among university students. Conclusion The proportion of self-reported hair loss was 44.61% among university students in Hangzhou City, and hair loss was predominantly mild. A family history of hair loss, nervousness/anxiety, diet habits, smoking and frequency of perm/dyeing hair may affect hair loss among university students.

Objective:To establish autoverification rules for coagulation tests in multicenter cooperative units, in order to reduce workload for manual review of suspected results and shorten turnaround time (TAT) of test reports, while ensure the accuracy of results.Methods:A total of 14 394 blood samples were collected from fourteen hospitals during December 2019 to March 2020. These samples included: Rules Establishment Group 11 230 cases, including 1 182 cases for Delta check rules; Rules Validation Group 3 164 cases, including 487cases for Delta check; Clinical Application Trial Group 77 269 cases. Samples were analyzed for coagulation tests using Sysmex CS series automatic coagulation analyzers, and the clinical information, instrument parameters, test results, clinical diagnosis, medication history of anticoagulant and other relative results such as HCT, TG, TBIL, DBIL were summarized; on the basis of historical data, the 2.5 and 97.5 percentile of all data arranged from low to high were initially accumulated; on the basis of clinical suggestions, critical values and specific drug use as well as relative guidelines, autoverification rules and limits were established.The rules were then input into middleware, in which Stage I/Stage II validation was done. Positive coincidence, negative coincidence, false negative, false positive, autoverification pass rate, passing accuracy (coincidence of autoverification and manual verification) were calculated. Autoverification rules underwent trial application in coagulation results reports.Results:(1) The autoverification algorisms involve 33 rules regarding PT/INR, APTT, FBG, D-dimer, FDP,Delta check, reaction curve and sample abnormalities; (2)Autoverification Establishment Group showed autoverification pass rate was 68.42% (7 684/11 230), the false negative rate was 0%(0/11230), coincidence of autoverification and manual verification was 98.51%(11 063/11 230), in which positive coincidence and negative coincidence were respectively 30.09% (3 379/11 230) and 68.42%(7 684/11 230); Autoverification Validation Group showed autoverification pass rate was 60.37%(1 910/3 164), the false negative rate was 0%(0/11 230), coincidence of autoverification and manual verification was 97.79%(3 094/3 164), in which positive coincidence and negative coincidence were respectively 37.42%(1 184/3 164) and 60.37%(1 910/3 164); (3) Trialed implementation of these autoverification rules on 77 269 coagulation samples showed that the average TAT shortened by 8.5 min-83.1 min.Conclusions:This study established 33 autoverification rules in coagulation tests. Validation showedthese rules could ensure test quality while shortening TAT and lighten manual workload.

Objective: To explore the effects of ambient fine particulate matter（PM2.5）exposure on semen quality in healthy adult men . Methods: The semen quality data were collected from the pre-pregnancy check outpatient of Taizhou Central Hospital from August 2014 to December 2017,excluding the patients with any disease of reproductive system. The data of daily pollutant concentration of this area during the same period were also collected to estimate the exposures of PM2.5,PM10,SO2,NO2,O3 and CO during 90 days before each semen examination. Logistic regression model was applied to estimate the association between PM2.5 exposure and semen quality . Results: A total of 3 456 healthy adult men were included in this study. The median （interquartile range）of the total sperm count of the 3 456 subjects was 161.00（122.00）×106,and 94.40% of the subjects could reach the lower reference limit for sperm count. The median（interquartile range）of the sperm concentration was 22.20（18.80）×106/mL,and 69.50% of the subjects were qualified. The median（interquartile range）of the sperm vitality was 55.30%（23.37%）,and 44.10% of the subjects were qualified. The median（interquartile range）of daily average concentration of PM2.5 at 0-90 days before semen examination was 37.82（12.33）μg/m3. The results of multivariate logistic regression model showed that,with each 12.33 μg/m3 increase of PM2.5 exposures at 0-90 days before semen examination,the odds ratios of unqualified sperm concentration,sperm motility and sperm vitality were 1.236（95%CI：1.041-1.466）,1.493（95%CI：1.260-1.770）and 1.462（95%CI：1.246-1.715）,respectively . Conclusion The results suggested that high concentration of ambient PM2.5 exposure during the sperm development period negatively affects sperm concentration,sperm motility and sperm vitality.

Direct secretory expression of active microbial transglutaminase (MTG) using heterologous hosts is a promising strategy, although its production level still needs to be improved for industrial production. Pichia pastoris is one of the most efficient expression systems developed in recent years. In this study, secretory expression of active MTG was successfully achieved by co-expressing the pro sequence and mature MTG genes in P. pastoris. Furthermore, we optimized the copy number of pro/MTG expression cassettes and the fermentation conditions. MTG production level reached 7.3 U/mL in 1-liter fermentor through high density fermentation, providing the feasiblity for industrial scale preparation of MTG.
Fermentation ; Genetic Vectors ; genetics ; Pichia ; enzymology ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Streptomyces ; enzymology ; Transglutaminases ; biosynthesis ; genetics