ObjectiveTo investigate the mechanism of Forsythiae Fructus-Lonicerae Japonicae Flos（FF） in the treatment of acute lung injury（ALI） by investigating the effects of FF on serum metabolomics of rats with ALI. MethodsThirty male SD rats were acclimated for 1 week， and 6 rats were randomly selected as the blank group. The other 24 rats were injected with lipopolysaccharide（LPS） solution by tracheal drip to establish an ALI model. After successful model establishment， the rats were randomly divided into the model group， the FF low-dose group（3.0 g·kg^-1）， the FF high-dose group（6.0 g·kg^-1）， and the dexamethasone group（5 mg·kg^-1）， with six rats in each group. The FF low- and high-dose groups and the dexamethasone group were received daily oral administration of the corresponding drug solution， and the blank group and the model group were gavaged with an equal amount of saline， treatment was administered continuously for 3 d. The pathological conditions of rat lung tissues were evaluated by hematoxylin-eosin（HE） staining， wet/dry mass ratio（W/D） of the lung tissues， and protein concentration in rat bronchoalveolar lavage fluid（BALF）. Metabolomic analysis of rat serum was performed by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）， combined with multivariate statistical analysis， the potential biomarkers of FF in treating ALI were screened by variable importance in the projection（VIP） value>1， P<0.05 from t-test， and log₂fold change（FC）>1 or log₂FC<-1. Kyoto Encyclopedia of Genes and Genomes（KEGG） database combined with MetaboAnalyst were used for pathway analysis of the screened differential metabolites. The protein expression levels of sphingosine-1-phosphate（S1P）， phosphatidylinositol 3-kinase（PI3K）， protein kinase B1（Akt1）， and phosphorylated Akt1（p-Akt1） were examined by Western bolt. The expression levels of interleukin（IL）-6， IL-1β， and tumor necrosis factor（TNF）-α in BALF were detected by enzyme-linked immunosorbent assay（ELISA）. ResultsCompared with the blank group， rats in the model group showed ALI pathological features such as alveolar lumen dilatation， interstitial hemorrhage and massive inflammatory cell infiltration， and the protein concentration in BALF and W/D of the lung tissues were significantly elevated（P<0.01）. Compared with the model group， the low- and high-dose groups of FF as well as the dexamethasone group exhibited reduced pulmonary bronchial hemorrhage in rats， and the protein concentration in BALF and W/D were significantly decreased（P<0.05）， and the lung injury was significantly alleviated. Analysis of rat serum metabolomics revealed that FF downregulated 38 biomarkers. Pathway enrichment analysis showed that FF primarily exerted therapeutic effects through 7 key metabolic pathways， including arginine biosynthesis， sphingomyelin metabolism， alanine， aspartate and glutamate metabolism， taurine and hypotaurine metabolism， α-linolenic acid metabolism， niacin and nicotinamide metabolism， and retinol metabolism. The results of Western bolt and ELISA showed that， compared with the blank group， the model group exhibited significantly elevated expression levels of S1P， PI3K， Akt1 and p-Akt1 proteins in the lung tissues， as well as increased expression levels of IL-6， IL-1β and TNF-α in BALF（P<0.01）. Compared with the model group， the expression levels of the aforementioned indicators were significantly downregulated in the low- and high-dose FF groups as well as the dexamethasone group（P<0.05， P<0.01）. ConclusionFF may play a role in ALI by regulating amino acid metabolism and lipid metabolism， and its mechanism may be related to the inhibition of S1P/PI3K/Akt1 signaling pathway to attenuate the inflammatory response caused by ALI.

N⁶-methyladenosine (m⁶A) modification plays a critical role in cell cycle regulation, while the mechanism of m⁶A in regulating mitosis remains underexplored. Here, we found that the total m⁶A modification level in cells increased during mitosis by the liquid chromatography-mass spectrometry/mass spectrometry and m⁶A dot blot assays. Silencing methyltransferase-like 3 (METTL3) or METTL14 results in delayed mitosis, abnormal spindle assembly, and chromosome segregation defects by the immunofluorescence. By analyzing transcriptome-wide m⁶A targets in HeLa cells, we identified polo-like kinase 1 (PLK1) as a key gene modified by m⁶A in regulating mitosis. Specifically, through immunoblotting and RNA pulldown, m⁶A modification inhibits PLK1 translation via YTH N⁶-methyladenosine RNA binding protein 1, thus mediating cell cycle homeostasis. Demethylation of PLK1 mRNA leads to significant mitotic abnormalities. These findings highlight the critical role of m⁶A in regulating mitosis and the potential of m⁶A as a therapeutic target in proliferative diseases such as cancer.
Humans ; Polo-Like Kinase 1 ; Cell Cycle Proteins/metabolism* ; Proto-Oncogene Proteins/metabolism* ; Protein Serine-Threonine Kinases/metabolism* ; Mitosis/physiology* ; HeLa Cells ; Adenosine/genetics* ; Methyltransferases/metabolism* ; RNA, Messenger/metabolism* ; RNA-Binding Proteins/metabolism*

Objective:To analyze the risk factors for 6-month prognosis of traumatic spinal cord injury (TSCI).Methods:A retrospective analysis was conducted of the 133 patients with TSCI who had been admitted to Department of Orthopaedics, The First Hospital Affiliated to Jinan University from January 2017 to August 2021. The patients with TSCI were categorized into an improved group ( n=82) and a non-improved group ( n=51) according to the improvements in the American Spinal Injury Association (ASIA) grading between admission and 6 months after injury. To identify the risk factors that might affect the prognosis of TSCI patients at 6 months after injury, univariate and logistic regression analyses were conducted of indicators such as gender, age, length of MRI spinal cord signal change, maximum canal compromise (MCC), maximum spinal cord compression (MSCC), brain and spinal cord injury center(BASIC)score, neutrophil-to-lymphocyte ratio (NLR) within 3 days after injury, ASIA grading within 3 days after injury, mean arterial pressure (MAP) at 3 days after operation, and presence of complications. Results:The univariate analysis showed significant differences between the improved and non-improved groups in length of MRI spinal cord signal change, MCC, MSCC, BASIC, NLR within 3 days after injury, ASIA grading within 3 days after injury, MAP at 3 days after operation, and presence of complications (all P<0.05). The logistic regression analysis showed that NLR ( OR=0.463, 95% CI: 0.287 to 0.748, P=0.002) and ASIA grading ( OR=11.684, 95% CI: 1.684 to 81.086, P=0.013) within 3 days after injury, as well as MAP at 3 days after operation ( OR=2.224, 95% CI: 1.306 to 3.787, P=0.003), were risk factors affecting the 6-month prognosis in TSCI patients. Conclusion:The NLR and ASIA grading within 3 days after injury, and MAP at 3 days after operation are risk factors that may affect the prognosis of TSCI patients at 6 months after injury.

The maintenance of hematopoietic stem cells (HSCs) is a complex process involving numerous cell-extrinsic and -intrinsic regulators. The first member of the cyclin-dependent kinase family of inhibitors to be identified, p21, has been reported to perform a wide range of critical biological functions, including cell cycle regulation, transcription, differentiation, and so on. Given the previous inconsistent results regarding the functions of p21 in HSCs in a p21-knockout mouse model, we employed p21-tdTomato (tdT) mice to further elucidate its role in HSCs during homeostasis. The results showed that p21-tdT+ HSCs exhibited increased self-renewal capacity compared to p21-tdT- HSCs. Zbtb18, a transcriptional repressor, was upregulated in p21-tdT+ HSCs, and its knockdown significantly impaired the reconstitution capability of HSCs. Furthermore, p21 interacted with ZBTB18 to co-repress the expression of cKit in HSCs and thus regulated the self-renewal of HSCs. Our data provide novel insights into the physiological role and mechanisms of p21 in HSCs during homeostasis independent of its conventional role as a cell cycle inhibitor.
Animals ; Hematopoietic Stem Cells/cytology* ; Cyclin-Dependent Kinase Inhibitor p21/genetics* ; Mice ; Cell Self Renewal ; Repressor Proteins/genetics* ; Mice, Inbred C57BL ; Mice, Knockout ; Humans ; Gene Expression Regulation

Background Phenolic compounds may adversely affect human health, but the current relevant studies are mostly limited to the impact of single phenolic compound exposure on human health, and there is still a lack of studies on the population-based association between combined exposure to multiple common phenolic compounds and dyslipidemia. Objective To explore the association of phenolic compound combined exposure and dyslipidemia based on principal component analysis-random forest (PCA-RF) strategy. Methods The data were from the National Health and Nutrition Examination Survey (2013–2016). A total of 1301 adult residents aged ≥ 20 years with complete information on demographics and lifestyle, urine phenol concentrations (bisphenol A, bisphenol F, bisphenol S, triclocarban, benzophenone, and triclosan), and serum concentrations of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were included in this study. The concentrations of six urinary phenolic compounds were determined by solid phase extraction coupled with high performance liquid chromatography and tandem mass spectrometry, and the lipid indicators were determined by enzymatic methods. Principal component analysis combined with random forest model was used for model construction. First, principal component analysis was performed on 18 original variables including 6 phenolic compounds and 12 basic characteristic indicators, and then random forest model was established with dyslipidemia and its four evaluation indicators as dependent variables and the extracted principal components as independent variables, respectively. Results The PCA-RF analysis showed that bisphenol A, bisphenol F, and benzophenone may be important factors for dyslipidemia in the study subjects; bisphenol A, bisphenol F, and triclosan may be important factors for TC level in the study subjects; bisphenol A, bisphenol F, triclocarban, and benzophenone may be important factors for TG level in the study subjects; bisphenol A may be an important factor for LDL-C level in the study subjects; bisphenol F and benzophenone may be important factors for HDL-C level in the study subjects. Conclusion Phenolic compound exposure may be an important risk factor for the development of dyslipidemia. PCA-RF strategy can be effectively used to explore the association between phenolic compound exposure and dyslipidemia in the population.

Enterotoxigenic Escherichia coli（ETEC）infection can induce watery diarrhea，leading to dehydration，electrolyte disturbance，and even death in severe cases. Recombinant B subunit/inactivated whole-cell cholera（rBS/WC）vaccine is effective in preventing ETEC infectious diarrhea. On the basis of the latest evidence on etiology and epidemiology of ETEC，as well as the effectiveness，safety，and health economics of rBS/WC vaccine，National Clinical Research Center for Child Health（The Children’s Hospital，Zhejiang University School of Medicine）and Zhejiang Provincial Center for Disease Control and Prevention invited experts to develop expert consensus on rBS/WC vaccine in prevention of ETEC infectious diarrhea. It aims to provide the clinicians and vaccination professionals with guidelines on using rBS/WC vaccine to reduce the incidence of ETEC infectious diarrhea.

Objective:To study the risk factors of metabolic bone disease (MBD) associated fracture in very low birth weight premature infants.Methods:From January 2012 to December 2019, premature infants (gestational age <32 weeks, birth weight <1 500 g) were admitted to our hospital and followed-up regularly for 1.5 years (once every month within first 6 months, then once every 3 months). The infants were assigned into two groups according to X-ray diagnosis: the fracture group and the non-fracture group. The clinical data of the two groups were compared and the risk factors of fracture were analyzed.Results:A total of 62 preterm infants with MBD were included in this study, including 11 in the fracture group and 51 in the non-fracture group. The risk factors of MBD associated fracture included intrauterine growth restriction (IUGR), birth weight <1 000 g, gestational age, respiratory support duration and total parenteral nutrition (TPN) duration ( P<0.05). Logistic regression analysis showed that IUGR ( P<0.05, OR=2.159, 95% CI 1.536~2.759) and TPN duration ( P<0.05, OR=1.143, 95% CI 1.042~1.270) were independent risk factors for fracture. Serum alkaline phosphatase (ALP) in the fracture group was significantly higher than the non-fracture group and 25(OH)VitD was significantly lower than the non-fracture group ( P<0.05). Conclusions:IUGR and TPN duration are risk factors for MBD associated fracture in preterm infants. As biochemical markers of bone metabolism, ALP and 25(OH)VitD levels have clinical value predicting MBD associated fracture.

Objective:To compare the dosimetric difference between IMRT and VMAT plans for ovarian protection after cervical cancer ovarian transposition surgery.Methods:Thirty-one patients who had received both cervical cancer resection and ovarian transposition were selected for adjuvant radiotherapy. The 9-field evenly divided IMRT and the dual-arc VMAT technology were performed for the treatment planning. The difference of the ovarian mean dose between the two techniques was explored. The relationship between the position of the ovarian-target interval and the ovarian dose was also analyzed.Results:A total of 54 ovaries in 31 patients were effectively transposed and moved out of the target area. Among them, 9 ovaries were located above the upper boundary of the PTV. For these cases, the ovarian mean dose of IMRT and VMAT were (177.8±90.7) and (166.7±70.6) cGy, respectively, which was not statistically different( P>0.05).45 ovaries were located in the same level with PTV. For these cases, the ovarian mean dose of IMRT and VMAT were (459.1±239.9) and (428.3±238.2) cGy, respectively ( z=3.11, P=0.002). The ovarian mean dose has the highest correlation and negative correlation with the closest lateral distance from the ovarian volume center to the PTV surface (IMRT, r=-0.922, P=0.001; VMAT, r=-0.865, P=0.001). To reduce the ovarian mean dose to 500 cGy, the lateral closest distance between the ovarian volume center and the PTV surface should be 3.6 cm and 3.3 cm for IMRT and VMAT respectively. Conclusions:There is no difference between the two planned ovarian doses when the ovaries were located above the upper boundary of the PTV. When the ovaries were located in the same level with PTV, the VMAT plan is better than IMRT in both ovarian dose and treatment efficiency. The ovarian dose could be predicted by the lateral closest distance from the ovarian volume center to the PTV.

The motion of the tumor limits further improvement in the accuracy of radiotherapy. Real-time monitoring and tracking of tumor location is an emerging technology to improve the accuracy of tumor radiotherapy. According to the adopted methods, it can be broadly divided into non-radiation-based and radiation-based systems. The former system includes ultrasound guidance, nuclear magnetic resonance guidance, electromagnetic tracking, optical image guidance, artificial intelligence-based technologies, and the latter system consists of KV, MV-grade X-ray imaging system and CT-based guidance system. In this review, research progresses on real-time tumor monitoring and tracking technology in radiotherapy, respective advantages and disadvantages and current clinical application were summarized.

OBJECTIVE：To investigate the effects and its mechanism of pseudostrychnine on the apoptosis of human colon can - cer HT- 29 cells. METHODS ：Human colon cancer HT- 29 cells were randomly divided into blank group ，pseudostrychnine low-dose，medium-dose and high-dose groups （125，250，500 μmol/L）. They were cultured with culture medium without medicine or relevant concentration of pseudostrychnine for 48 h. The cell apoptosis and mitochondrial transmembrane potential were detected by flow cytometry. Western blotting assay was employed to detect the protein expression of P 53，Caspase-3，Caspase-9，DNA re - pair enzyme from rabbit （c-PARP）and Bcl- 2. RESULTS ：Compared with blank group ，apoptotic rate of cells was increased signifi - cantly in pseudostrychnine low-dose ，medium-dose and high-dose groups ，while mitochondrial transmembrane potential was de - creased significantly （P＜0.01），in concentration-dependent manner. The protein expression levels of P 53，Caspase-3，Caspase-9 and c-PARP were increased in pseudostrychnine medium-dose and high-dose groups ，compared with blank group ；while those of Bcl-2 were decreased significantly in pseudostrychnine low-dose ，medium-dose and high-dose groups （P＜0.05 or P＜0.01）. CON - CLUSIONS：Pseudostrychnine may change mitochondrial membrane potential by up-regulating the protein expression of P 53 and down-regulating the protein expression of Bcl- 2，and activate the expression of Caspase- 3，c-PARP and Caspase- 9，so as to acti - vate endogenous mitochondrial pathway and promote the apoptosis of HT- 29 cells.