ObjectiveTo investigate the effect and mechanism of Polygonatum neutral polysaccharides from sibiricum （PSP-NP） on colon injury in mice with chronic obstructive pulmonary disease （COPD）. MethodsMale C57BL/6J mice were randomly divided into a control group， a COPD model group， and a PSP-NP group. The COPD model was established using smoke exposure combined with intranasal LPS administration. The PSP-NP group was simultaneously treated daily with 200 mg/kg of PSP-NP via intragastric gavage， while the other groups received an equal volume of saline. HE staining was used to observe the pathological changes in the colon. ELISA was employed to detect the levels of LPS in serum and the expressions of ZO-1， Occludin， IL-6， and TNF-α in colon tissue. UPLC-MS was used to detect the types and contents of bile acids in colonic content， and to screen for differential bile acids. Differential microbial flora were identified using 16S rRNA gene sequencing， and correlation analysis was conducted with differential bile acids. PSP-NP was combined with the differential bile acids cholic acid （CA）， and deoxycholic acid （DCA） in vitro to analyze the binding capacity of PSP-NP for CA and DCA. PSP-NP was applied to NCM460 normal colonic epithelial cells cultured in CA and DCA. Cell migration ability was assessed using the scratch assay， and the mRNA expression levels of inflammatory cytokines TNF-α， IL-6， and NF-κB were measured by RT-qPCR. ResultsPSP-NP effectively improved colonic damage in COPD model mice， enhanced mechanical barrier function， alleviated inflammatory response， and regulated abnormal changes in colonic flora and bile acid metabolism. Correlation analysis further revealed that PSP-NP regulated colonic bile acid metabolism and reduced the redundancy of secondary bile acids by increasing the relative abundance of Bacteroidota， Verrucomicrobiota， Bacteroides， and Akkermansia， while decreasing the relative abundance of Lactobacillus and Bifidobacterium. Notably， in vitro binding assays demonstrated that PSP-NP bound to differential bile acids DCA and CA， with the strongest binding capacity for DCA at 58.2%. In cellular functional studies， DCA inhibited the migration ability of colonic epithelial cells NCM460 and significantly increased the relative mRNA expression levels of inflammatory factors TNF-α， IL-6， and NF-κB. Importantly， co-treatment with PSP-NP significantly ameliorated the impact of DCA on NCM460 cells. ConclusionsPSP-NP may significantly improve colonic damage in COPD model mice. The mechanism may involve the regulation of colonic bile acid metabolism and bile acid profiles through both microbial modulation and direct binding， thereby reducing the damage caused by secondary bile acids such as DCA to colonic epithelial cells.

ObjectiveTo investigate the pharmacodynamic effects of Houshihei San （HSHS） recorded with the effects of treating wind and limb heaviness on muscle tissue injury after middle cerebral artery occlusion （MCAO） in rats through the ferroptosis pathway. MethodsThirty SD male rats were selected and randomly grouped as follows： sham， MCAO， deferoxamine mesylate， high-dose HSHS （HSHS-H， 0.54 g·kg^-1）， and low-dose HSHS （HSHS-L， 0.27 g·kg^-1）， with 6 rats in each group. A laser scattering system was used to evaluate the stability of the MCAO model， and rats were administrated with corresponding agents by gavage for 7 days. During the administration period， behavioral， imaging and other methods were used to systematically evaluate the skeletal muscle tissue injury after MCAO and the therapeutic effect in each administration group. Hematoxylin-eosin staining was employed to evaluate the cross-section of muscle cells. Subsequently， immunohistochemistry was used to detect tumor suppressor p53 and glutathione peroxidase 4 （GPX4） in the soleus tissue. Western blot was employed to determine the protein levels of p53， GPX4， myogenic differentiation 1 （MyoD1）， nuclear factor E₂-related factor 2 （Nrf2）， Myostatin， solute carrier family 7 member 11 （SLC7A11）， muscle ring-finger protein-1 （MuRF1）， and muscle atrophy F-box protein （MAFbx） to verify the therapeutic effect in each group. ResultsCompared with the MCAO group， HSHS enhanced the locomotor ability and promoted muscle regeneration， which suggested that the pharmacological effects of HSHS were related to the inhibition of muscle tissue ferroptosis to reduce the expression of muscle atrophy factors. Behavioral and imaging results suggested that compared with the MCAO group， HSHS ameliorated neurological impairments in rats on day 7 （P<0.01）， enhanced 5-min locomotor distance and postural control （P<0.01）， strengthened grasping power and promoted muscle growth （P<0.01）， stabilized skeletal muscle length and weight （P<0.01）， and increased the cross-section of muscle cells （P<0.01）. Compared with the MCAO group， HSHS promoted the increases in glutathione and superoxide dismutase content and inhibited the increase in malondialdehyde content （P<0.05，P<0.01）. Ferroptosis pathway-related assays suggested that HSHS reduced the p53-positive cells and increased the GPX4-positive cells （P<0.01）. HSHS ameliorated muscle function decline after stroke by promoting the expression of GPX4， Nrf2， SLC7A11， and MyoD1 and inhibiting the expression of p53， Myostatin， MurRF1， and MAFbx to reduce ferroptosis in the muscle （P<0.01）. ConclusionHSHS， prepared with reference to the method in the Synopsis of Golden Chamber， can simultaneously reduce the myolysis and increase the protein synthesis in the skeletal muscle tissue after ischemic stroke by regulating the ferroptosis pathway.

ObjectiveThis study aims to investigate the inhibitory effect of Wutoutang on pannus formation in adjuvant-induced arthritis （AIA） rats with wind-cold-dampness Bi syndrome and its potential mechanism. MethodA total of 40 male SD specific pathogen-free （SPF） rats were selected and divided into blank group， wind-cold-dampness Bi syndrome group ［Complete Freund's Adjuvant （CFA）， 200 μg］， Wutoutang group （15 g·kg^-1·d^-1）， and indometacin group （10 mg·kg^-1） according to random number table method. Except for the blank group， the other groups were given wind-cold-dampness stimulation before the CFA injection. After the rats were administered for 30 days， the basic conditions， onset time， arthritis index score， and foot swelling volume of AIA rats with wind-cold-dampness Bi syndrome were observed. Finally， peripheral arterial blood， ankle joint， and synovial tissue were taken. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum hypoxia-inducible factor-1α （HIF-1α）， vascular endothelial growth factor A （VEGFA） protein content， and rheumatism， including anti-O （ASO）， C-reactive protein （CRP）， and rheumatoid factor （RF）. Hematoxylin-eosin （HE） staining revealed the changes in joint histomorphology. Immunohistochemistry was used to detect the expression of HIF-1α and VEGFA， two important proteins in the ankle pathway. Quantitative real-time polymerase chain reaction （Real-time PCR） was used to reveal mRNA levels of HIF-1α， VEGFA， angiopoietin-1 （Ang-1）， and angiopoietin-2 （Ang-2） in rat synovial tissue. ResultThe foot swelling volume and arthritis score of AIA rats with wind-cold-dampness Bi syndrome were substantially higher （P<0.01） compared with the blank group. Serum CRP， RF， and ASO levels were considerably elevated （P<0.01）. HE staining showed obvious hyperplasia of ankle synovium and synovial inflammation， angiogenesis and pannus formation， and aggravated bone destruction， indicating successful modeling. After the intervention of Wutoutang， the onset time was delayed （P<0.01）. Foot swelling volume and arthritis score were decreased （P<0.01）. Serum CRP， RF， and ASO levels were significantly decreased （P<0.01）. The inflammatory hyperplasia of synovial tissue， angiogenesis and pannus formation， and bone destruction were alleviated. The mRNA levels of HIF-1α， VEGFA， Ang-1， and Ang-2 in the synovial membrane were significantly decreased （P<0.05， P<0.01）. The expressions of HIF-1α and VEGFA in serum and ankle joints were decreased （P<0.01）. In the indomethacin group， the onset time of the disease was delayed （P<0.01）. Foot swelling volume and arthritis score were decreased （P<0.01）. Serum CRP， RF， and ASO levels were significantly decreased （P<0.01）. HIF-1α/VEGFA/Ang signaling pathway was activated， and pathological tissue injury was improved. ConclusionWutoutang can delay the onset time of AIA rats with wind-cold-dampness Bi syndrome， reduce foot swelling volume， arthritis score， rheumatic activity， and improve joint histopathology. It can inhibit pannus formation， and its mechanism may be related to down-regulating the expression of the HIF-1α/VEGFA/Ang pathway.

Objective To understand the viral spectrum of inpatients with acute respiratory infection in Pudong New Area, and to explore the composition of pathogens in hospitalized children and adults. Methods Samples of acute respiratory infection cases from 10 medical institutions were collected from 2011 to 2020 and tested for human influenza virus, human adenovirus, rhinovirus, human parainfluenza virus, respiratory syncytial virus, human coronavirus, human metapneumovirus and human boca virus. Results A total of 3 145 inpatients were monitored, with a median age of 61 years. The positive rate of any virus was 32.43% (1 020/3 145), and the single virus infection accounted for 85.98% (877/1 020). In single virus infection, the positive rate of human influenza virus was the highest (9.67%, 304/3 145), with influenza A (80.26%, 244/304) as the main virus. The second was rhinovirus (3.97%, 125/3 145). The positive rate of any virus in different age groups was statistically significant (χ²=103.38，P<0.001). The positive rate of respiratory syncytial virus was the highest in the ~5-year-old group, adenovirus was the highest in the 6-14-year-old group, and influenza virus was the highest in the 15-64-year-old group and the 65year-old group. There was a significant difference in the positive rate of any virus in each month (χ²=123.06，P<0.001). The human influenza virus was the dominant virus in winter (December to February) and summer (July to September), and rhinoviruses distributed sporadically in each month. The positive rate of any virus in different departments was significantly different (χ²=90.37，P<0.001). Conclusion The positive rate of virus in hospitalized patients with acute respiratory infection is relatively high in Pudong New Area, Shanghai, with human influenza virus being the main virus. The virus spectrum of hospitalized children and adults is inconsistent. In the future, in-depth research should be strengthened, focusing on the distribution of pathogens in different populations and seasonal prevention and treatment.

OBJECTIVE To explore the effects of alfentanil （ALF） on myocardial fibrosis in rats with acute myocardial infarction （AMI） by regulating sphingosine kinase 1 （SphK1）/sphingosine 1-phosphate （S1P） signaling pathway. METHODS Male SD rats were collected to construct AMI model by the ligation of anterior descending branch of left coronary artery. The successfully modeled rats were randomly divided into AMI model group （Model group）， ALF low-dose group （ALF-L group， 0.25 mg/kg ALF）， ALF high-dose group （ALF-H group， 0.5 mg/kg ALF）， high dose of ALF+SphK1 activator group （ALF-H+K6PC-5 group， 0.5 mg/kg ALF+1 μg/g K6PC-5）. At the same time， a sham operation group （Sham group） was set up to perform only chest opening/closing operations without ligating the anterior descending branch of left coronary artery， with 15 rats in each group. Rats in each drug group were intraperitoneally injected with the corresponding drug solution， once a day， for 4 consecutive weeks. Twelve hours after the last medication， cardiac function indicators [left ventricular systolic pressure （LVSP）， left ventricular ejection fraction （LVEF）， left ventricular systolic diameter （LVSD）， left ventricular fractional shortening （LVFS）] of rats were detected in each group； the condition of myocardial infarction， pathological changes in myocardial tissue， and degree of fibrosis were observed； serum levels of brain natriuretic peptide （BNP） and cardiac troponin Ⅰ （cTnⅠ） in rats were detected. The protein expressions of collagen Ⅰ ， collagen Ⅲ ， matrix metalloproteinase-2 （MMP-2）， SphK1 and S1P were alsodetected in the myocardial tissue of rats. RESULTS Compared with the Sham group， the arrangement of myocardial cells in the Model group was disordered， with a large number of inflammatory cells infiltrating. The levels of LVSP， LVFS and LVEF in the Model group were significantly reduced （P＜0.05）； LVSD level， myocardial infarction area， collagen volume fraction， serum levels of BNP and cTnⅠ， the protein expressions of collagen Ⅰ， collagen Ⅲ， MMP-2， SphK1 and S1P in myocardial tissue were significantly increased or enlarged （P＜0.05）. Compared with the Model group， the pathological changes and degree of fibrosis in the myocardial tissue of rats in each dose group of ALF were improved or relieved， while the quantitative indicators of rats in the ALF-H group were significantly improved and significantly better than those in ALF-L group （P＜0.05）. K6PC-5 could significantly reverse the improvement effect of high-dose ALF on the above quantitative indicators in rats （P＜0.05）. CONCLUSIONS ALF can reduce myocardial fibrosis and improve cardiac function in AMI rats， and the effect may be related to the inhibition of the SphK1/S1P signaling pathway.

Endo-beta-N-acetylglucosaminidase (ENGase) is widely distributed in various organisms. The first reported ENGase activity was detected in Diplococcus pneumoniae in 1971. The protein (Endo D) was purified and its peptide sequence was determined in 1974. Three ENGases (Endo F1-F3) were discovered in Flavobacterium meningosepticum from 1982 to 1993. After that, the activity was detected from different species of bacteria, yeast, fungal, plant, mice, human, etc. Multiple ENGases were detected in some species, such as Arabidopsis thaliana and Trichoderma atroviride. The first preliminary crystallographic analysis of ENGase was conducted in 1994. But to date, only a few ENGases structures have been obtained, and the structure of human ENGase is still missing. The currently identified ENGases were distributed in the GH18 or GH85 families in Carbohydrate-Active enZyme (CAZy) database. GH18 ENGase only has hydrolytic activity, but GH85 ENGase has both hydrolytic and transglycosylation activity. Although ENGases of the two families have similar (β/α)8-TIM barrel structures, the active sites are slightly different. ENGase is an effective tool for glycan detection andglycan editing. Biochemically, ENGase can specifically hydrolyze β‑1,4 glycosidic bond between the twoN-acetylglucosamines (GlcNAc) on core pentasaccharide presented on glycopeptides and/or glycoproteins. Different ENGases may have different substrate specificity. The hydrolysis products are oligosaccharide chains and a GlcNAc or glycopeptides or glycoproteins with a GlcNAc. Conditionally, it can use the two products to produce a new glycopeptides or glycoprotein. Although ENGase is a common presentation in cell, its biological function remains unclear. Accumulated evidences demonstrated that ENGase is a none essential gene for living and a key regulator for differentiation. No ENGase gene was detected in the genomes of Saccharomyces cerevisiae and three other yeast species. Its expression was extremely low in lung. As glycoproteins are not produced by prokaryotic cells, a role for nutrition and/or microbial-host interaction was predicted for bacterium produced enzymes. In the embryonic lethality phenotype of the Ngly1-deficient mice can be partially rescued by Engase knockout, suggesting down regulation of Engase might be a solution for stress induced adaptation. Potential impacts of ENGase regulation on health and disease were presented. Rabeprazole, a drug used for stomach pain as a proton inhibitor, was identified as an inhibitor for ENGase. ENGases have been applied in vitro to produce antibodies with a designated glycan. The two step reactions were achieved by a pair of ENGase dominated for hydrolysis of substrate glycoprotein and synthesis of new glycoprotein with a free glycan of designed structure, respectively. In addition, ENGase was also been used in cell surface glycan editing. New application scenarios and new detection methods for glycobiological engineering are quickly opened up by the two functions of ENGase, especially in antibody remodeling and antibody drug conjugates. The discovery, distribution, structure property, enzymatic characteristics and recent researches in topical model organisms of ENGase were reviewed in this paper. Possible biological functions and mechanisms of ENGase, including differentiation, digestion of glycoproteins for nutrition and stress responding were hypothesised. In addition, the role of ENGase in glycan editing and synthetic biology was discussed. We hope this paper may provide insights for ENGase research and lay a solid foundation for applied and translational glycomics.

Objective To systematically evaluate the efficacy and safety of the combination therapy of sodium valproate and edaravone in the treatment of post-stroke epilepsy(PSE).Methods The Cochrane Library,PubMed,Embase,Web of Science,CNKI,Wanfang database,and VIP databases were searched from the time of inception to May 2023.Randomized controlled trials of edaravone plus valproate versus valproate monotherapy for epilepsy were collected,the risk of bias of the included studies was assessed using the RoB 2 tool,and Meta-analyses were performed using RevMan 5.4 software.Results 13 studies with a total of 1 092 patients were included.The results showed that the response rate was significantly higher in the combination group than in the sodium valproate group[RR=0.18,95％CI(0.13 to 0.22),P＜0.01].The incidence of adverse reactions was lower[RR=0.73,95％CI(0.48 to 1.13),P=0.16].The seizure frequency[MD=-0.30,95％CI(-0.43,-0.11),P＜0.01]and duration of seizures[MD=-0.81,95％CI(-0.89,-0.72),P＜0.01]in the combined treatment group were significantly lower than those in the sodium valproate group.The inflammatory factors tumor necrosis factor-α[MD=-8.00,95％CI(-9.15,-6.84),P＜0.01],interleukin-2[MD=-10.19,95％CI(-14.61,-5.78),P＜0.01],interleukin-8[MD=-5.6,95％CI(-6.48,-4.73),P＜0.01]were significantly lower than those in the sodium valproate group.The neuron-specific enolase(NSE)level in patients with epilepsy in the combination treatment group was significantly better than that of the sodium valproate group at 1 month[MD=-4.73,95％CI(-4.99,-4.46),P＜0.01],3 months[MD=-2.10,95％CI(-3.26,-0.95),P＜0.01],and 6 months after treatment[MD=-1.31,95％CI(-2.35,-0.27),P＜0.01].There was no significant difference in NSE levels[MD=0.06,95％CI(-0.07,0.19),P=0.34]after 12 months.Conclusion Edaravone combined with sodium valproate in the treatment of post-stroke epilepsy can improve the epilepsy control rate of patients,shorten the frequency and duration of seizures,reduce the level of inflammatory factors in patients,promote the reduction of NSE levels,and improve their quality of life,which has a certain safety.

Objective: To understand current status and related factors of breakfast among primary and secondary school students in Zhejiang Province, so as to provide a scientific basis for improving breakfast habits of primary and secondary school students. Methods: During May to November of 2023, 33 326 students from grade four to six of primary schools and grade one to two of secondary schools were selected from 90 counties and cities in Zhejiang Province by using the stratified cluster random sampling method. General information and breakfast consumption were collected by questionnaire. Multivariate Logistic regression was used to analyze the related factors of breakfast. Results: About 81.29% of the primary and secondary school students reported regular breakfast consumption. The rate of regular breakfast consumption was higher on the school days (92.23%) than on the weekends (85.17%), and higher in primary school students (85.83%) compared to secondary school students (74.71%), with statistically significant differences ( χ 2=827.42, 655.03, P <0.01). About 49.19% of primary and secondary school students had their breakfast within 10 minutes or less, and 83.30% of primary and secondary school students had 3-5 food groups for breakfast. The proportions of students who consumed cereals and potatoes, milk, and eggs were respectively 18.76%, 28.85%, 14.63%. About 22.84%, 28.00 %, 32.60% and 32.23% of the students had no meat, soybeans, vegetables and fruits in their breakfast. The results of multivariate Logistic regression analysis showed that girls, rural area, secondary school, place of living (dormitory, others), migrant parent (one or both outside the hometown), late bedtime (22:00-22:59, 23:00 and later) and late wake up time (9:00 and later) on the weekends were positively correlated with no having breakfast every day ( OR=1.22, 1.40, 1.46, 1.20, 1.20, 1.34, 1.36, 1.41 , 3.51, 2.32, P <0.05). The time of physical activity per day (30-<60, 60-<90, 90-120, >120 min), bedtime (21:00-21:59, 22:00-22:59) and wake up time (6:00-6:59, 7:00-7:59) on school days were negatively correlated with no having breakfast every day ( OR=0.75, 0.64, 0.67, 0.64, 0.77, 0.82, 0.75, 0.67, P <0.05). Conclusions There is a considerable number of primary and secondary school students with irregular breakfast consumption, which are related to multiple factors. Therefore, it is necessary to strengthen nutrition education and improve the behavior of breakfast for primary and secondary school students.

ObjectiveTo understand the epidemiological characteristics of recovered COVID-19 cases with re-positive nucleic acid in Pudong New Area of Shanghai, and to provide a reference for the prevention and control of COVID-19. MethodsA three-month health follow-up and nucleic acid testing were conducted on 339 COVID-19 cases cured and discharged between May 20 and June 20, 2022, in Pudong New Area, Shanghai, to analyze their epidemiological characteristics. ResultsAmong the 339 follow-up cases, 75 cases experienced re-positive nucleic acid results, with a recurrence rate of 22.12%. Factors such as gender, age group, occupation, presence of heart disease, hypertension, and full vaccination status had no effect on the re-positive results. Being diagnosed as a confirmed case during the first presence of infection, having diabetes, and a hospitalization period of ≤7 days were related factors for recurrence. The median interval between discharge and re-positive nucleic acid results was 26 days. The close contacts of the re-positive cases did not contract COVID-19 after the isolation and observation period. ConclusionThere is a possibility of re-positive nucleic acid results after COVID-19 recovery and discharge. Cases initially diagnosed as confirmed cases and those with a hospitalization period of no more than 7 days have a high rate of re-positivity. No secondary transmission is observed from the re-positive cases.

AIM:To investigate the expression of centromere protein-H(CENP-H)in adrenocortical carcino-ma(ACC)and its relationship with disease progression and prognosis,and to explore the impact of CENP-H gene knock-down on the viability and migration of ACC cells.METHODS:The mRNA expression level of CENP-H in 76 ACC pa-tients and 128 healthy controls,and its correlations with tumor stages and prognosis were analyzed by GEPIA2 database.The mRNA expression of CENP-H in different stages of ACC and its correlation with disease prognosis were further ana-lyzed by ULCAN database.The protein expression of CENP-H was examined by immunohistochemical staining of paraffin-embedded ACC and normal adrenal gland specimens.Knockdown of CENP-H by siRNA(siCENP-H)was performed in human ACC cell line H295R.The viabilty of H295R cells transfected with siCENP-H or siNC was measured by CCK-8 as-say,the cell migration was detected by wound-healing assay,and the protein levels of CENP-H,p-ERK1/2,t-ERK1/2,p-P38,t-P38,p-JNK1/2 and t-JNK1/2 were detected by Western blot.RESULTS:The mRNA level of CENP-H was signifi-cantly higher in ACC than that in normal controls,and was correlated with tumor stages and prognosis.The protein level of CENP-H was significantly higher in ACC specimens than that in normal adrenal gland.Knockdown of CENP-H in H295R cells resulted in decreased cell viability and migration.The protein levels of p-P38 and p-JNK1/2 were decreased in si-CENP-H group.CONCLUSION:CENP-H is highly expressed in ACC,and is correlated with tumor stages and poor prognosis.Knockdown of CENP-H can inhibit the viability and migration of ACC cells,and its mechanism may related to inactivation of P38 and JNK signaling pathways.