ObjectiveTo reveal the active substances and mechanisms of modified Qing'e formula （MQEF） in delaying skin photoaging by ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry （UPLC-Q-TOF-MS），network pharmacology， and cell experiments. MethodsUPLC-Q-TOF-MS and a literature review were employed to analyze the transdermally absorbed components in mice after the topical application of MQEF. The potential targets of MQEF in treating skin photoaging were retrieved from databases.The compound-potential target network and protein-protein interaction network were constructed to screen the key components and core targets. A photoaging cell model was established by irradiating HaCaT cells with medium-wave ultraviolet B （UVB）. The safe doses of bakuchiol （BAK） and salvianolic acid B （SAB） for treating HaCaT cells and the effects of BAK and SAB on the viability of cells exposed to UVB irradiation were determined by the cell counting kit-8 （CCK-8） method.The reactive oxygen species （ROS） fluorescent probe was used to measure the ROS production in the cells treated with BAK and SAB.The expression levels of genes related to oxidative stress，inflammation，collagen metabolism，and circadian rhythm clock were measured by Real-time PCR. ResultsA total of 24 transdermally absorbed components of MQEF were identified，which acted on 367 potential targets，and 417 targets related to skin photoaging were screened out，among which 47 common targets were predicted as the targets of MQEF in treating skin photoaging. MQEF exerted the anti-photoaging effect via key components such as BAK and SAB，which acted on core proteins such as serine/threonine kinase 1 （Akt1） and mitogen-activated protein kinase 3 （MAPK3） and intervened in core pathways such as the tumor necrosis factor （TNF） and phosphatidylinositol-3-kinase （PI3K）/protein kinase B （Akt） signaling pathways.Compared with the model group，the administration of BAK and SAB increased the survival rate of HaCaT cells （P<0.01），down-regulated the mRNA levels of cyclooxygenase-2 （COX-2），interleukin-6 （IL-6），tumor necrosis factor-α （TNF-α），matrix metalloproteinase-1 （MMP-1），and matrix metalloproteinase-9 （MMP-9） （P<0.01），and up-regulated the mRNA levels of heme oxygenase-1 （HO-1） and NAD（P）H quinone dehydrogenase 1 （NQO-1） （P<0.05，P<0.01） in photoaged HaCaT cells.In addition，it eliminated excess ROS production induced by UVB and up-regulated the mRNA levels of brain and muscle ARNT-like 1 （BMAL1） and circadian locomotor output cycles kaput （CLOCK） associated with circadian clock （P<0.05，P<0.01）. ConclusionMQEF delays skin photoaging through the coordinated effects of various components，multiple targets，and diverse pathways.The key components BAK and SAB in MQEF exhibit anti-photoaging properties，which involve inhibiting oxidative stress，preventing collagen degradation，mitigating inflammation，and maintaining normal skin circadian rhythms by regulating clock gene expression.

BACKGROUND:Premature birth is a major global health problem associated with high mortality and morbidity.White matter injury is the most common brain injury in preterm infants.Salvia miltiorrhiza is a traditional herbal plant that is commonly used to treat cardiovascular and cerebrovascular diseases in Asian countries. OBJECTIVE:To investigate the therapeutic effect of Salvia miltiorrhiza on white matter injury in preterm infants. METHODS:Eighteen neonatal male Sprague-Dawley rats at 3-day gestational age were selected and randomized into normal group,white matter injury group,and Salvia miltiorrhiza group.Animal models of preterm white matter injury were established by permanent ligation of the right common carotid artery in the latter two groups.Rats in the Salvia miltiorrhiza group were given intraperitoneal injection of Salvia miltiorrhiza(5 mg/kg·d)for 7 consecutive days.Normal group and white matter injury group were given the same volume of PBS for intervention.On the 14th day after modeling,the rats were sacrificed.Brains were pathologically observed by hematoxylin-eosin staining under microscope,and the expression levels of myelin basic protein and CC1 in brain tissue were visualized using immunofluorescence.Furthermore,liquid chromatography-tandem mass spectrometry was used to analyze possible pathways for the action of Salvia miltiorrhiza. RESULTS AND CONCLUSION:In the white matter injury group,the structure of the corpus callosum was irregular and the cells appeared swollen and necrotic.In addition,induction of white matter injury resulted in significantly reduced myelin formation,with irregular and loosely arranged nerve fibers and significantly decreased myelin sheaths.Interestingly,white matter injury rats treated with Salvia miltiorrhiza had reduced cellular swelling,reduced lesions,and increased myelin sheaths.The expression of myelin basic protein was closely related to myelin formation,and CC1 was a marker of myelin oligodendrocytes.Salvia miltiorrhiza significantly up-regulated the expressions of myelin basic protein and CC1 in white matter injury rats(P＜0.000 1),indicating that Salvia miltiorrhiza alleviated white matter injury.Liquid chromatography-tandem mass spectrometry analysis showed that the therapeutic effect of Salvia miltiorrhiza in the rat model of white matter injury was closely related to the regulation of complement and coagulation cascades.To conclude,Salvia miltiorrhiza may be a potential therapeutic agent for treating preterm white matter injury.

Abstract: Objective　To compare and analyze the clinical characteristics, diagnosis, and treatment of different subtypes of severe influenza in neonates to provide a reference for the diagnosis and treatment of neonatal severe influenza. Methods A cohort of 40 neonates with severe influenza who were hospitalized in the neonatology ward of Children's Hospital Affiliated to Zhengzhou University between January 2019 to December 2023 were selected and divided into two groups based on the virus subtype: influenza A (n=23) and influenza B (n=17). A retrospective analysis was conducted to compare general information, clinical manifestations, auxiliary examinations, complications, and treatment outcomes of neonates with severe influenza A and B infection. Results The number of days of hospitalization was longer in cases of influenza A than that of influenza B. The proportion of neonates with severe influenza A who exhibited fever was higher than that for influenza B, and a higher percentage of those with fever had peak temperatures ranging from 38.1 ℃ to 39 ℃. Gastrointestinal symptoms, including vomiting and diarrhea leading to dehydration, were more evident in severe influenza B cases. The proportion of influenza A cases with abnormal creatine kinase-MB isoenzyme levels (>25 U/L) was higher than that of influenza B, and the differences were statistically significant (P<0.05). There were no significant differences between the two types of influenza in other clinical manifestations, the incidence of pneumonia/respiratory failure complications, peripheral blood leukocyte count and classifications, the proportion of abnormal aspartate aminotransferase (AST) (>40 U/L), alanine aminotransferase (ALT) (>40 U/L), and creatine kinase (CK) (>200 U/L), and lactate dehydrogenase (LDH) values (all P>0.05). In terms of treatment, neonates treated with Oseltamivir within 48 hours of onset mainly suffered from influenza A. Among those treated with Oseltamivir, the proportion of influenza A cases whose body temperature returned to normal within 24 hours was relatively higher, whereas, for those whose temperature returned to normal within 24-72 hours, the proportion was relatively higher in influenza B cases. These differences were statistically significant (all P<0.05). Conclusions Severe neonatal influenza usually occurs in winter and spring. After severe infection, fever is more obvious in neonates with influenza A, which is more likely to cause myocardial cell damage. Neonates with influenza A can be treated with Oseltamivir earlier and return to normal body temperature faster than those with influenza B after Oseltamivir treatment. Gastrointestinal symptoms are more common in neonates with severe influenza B.

Mycoplasma pneumoniae pneumonia is an important component of community-acquired pneumonia, which can cause severe extrapulmonary complications of digestive, cardiovascular, blood, urinary and other systems.Accurate and effective biomarkers are significant for the diagnosis and treatment of Mycoplasma pneumoniae pneumonia.Recent studies have shown that new biomarkers such as IL-35, IL-17, neutrophil to lymphocyte ratio(NLR) and S100 protein are involved in the development of Mycoplasma pneumoniae pneumonia.In order to provide reference for the clinical diagnosis and treatment of Mycoplasma pneumoniae pneumonia, this paper reviews the progress of new biomarkers in Mycoplasma pneumoniae pneumonia.

Objective:To investigate the effects of pregnancy and lactation nonylphenol (NP) exposure on the balance of Treg/Th17 cells in the brain of offspring mice and the related mechanisms.Methods:Thirty pregnant C57BL/6 mice were randomly divided into three groups: control group (drinking distilled water), and NP-treated groups (drinking 0.2 μg/ml or 2.0 μg/ml NP water solution). ELISA kit was used to analyze the levels of TNF-α, IFN-γ and IL-17, flow cytometry was used to analyze the frequency of Treg and Th17 cells in spleen, quantitative RT-PCR was used to analyze the RORγt, Foxp3 mRNA, Western blot was used to analyze the protein expression of RORγt, Foxp3 and PI3K/Akt/mTOR signal pathway, and immunofluorescence was used to analyze the expression of Iba1 in the brain tissue of offspring mice.Results:Compared with the control group, NP exposure increased the serum levels of IL-17 and TNF-α in male offspring mice ( P<0.05), and decreased the levels of IFN-γ( P<0.05). Flow cytometry analysis showed that the percentage of Th17 cells in the spleen of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) was significantly higher than that of the control group, while the percentage of Tregs cells was lower. Compared with the control group, the expression levels of Foxp3 proteins in the brain tissue of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) was significantly lower, accompanied by a dramatic increase in RORγt protein levels ( P<0.05). Similar mRNA expression was also observed in qRT-PCR analysis. The protein expression levels of mTOR (p-mTOR) and its upstream related regulators[PI3K, p-Akt (Ser473), p-Akt (Thr308)] in the brain of male offspring mice increased gradually during the period of exposure to NP( P<0.05). Immunofluorescence analysis showed that compared with the control group, the number of Iba1 positive cells in brain tissue of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) increased significantly ( P<0.05). Conclusions:Maternal exposure to NP during pregnancy and lactation may affect the development/function of neurons in offspring through neuroimmune axis, and increase the risk of neurodevelopmental disorders in offspring.

Objective:To explore the role and molecular mechanism of trophoblastic cell surface antigen 2 (Trop2) in the invasion and migration of ovarian cancer.Methods:Through the data mining of Cancer Cell Line Encyclopedia and TCGA database, the clinical significance of Trop2 expression was analyzed. Western blot was used to detect Trop2 protein expression in ovarian cancer cell lines including A3O, A1780 and SKOV3. SKOV3 cells were used to construct Trop2-short hairpin RNA (shRNA) cell model. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the SKOV3 mRNA expression in SKOV3-shRNA and SKOV3-NC cells. Cell counting kit-8 (CCK8) was used to detect the proliferation of SKOV3-shRNA cells and SKOV3-NC cells. Flow cytometry was used to detect cell cycle and apoptosis in two groups of cells. Transwell array was used to detecte the invasion and migration of SKOV3-shRNA cells and SKOV3-NC cells. Western blot was used to detect the protein expressions of AKT, p-AKT, β-catenin, caspase3, bcl-2, E-cadherin and vimentin.Results:Trop2 mRNA highly expressed in ovarian cancer, and was related to the tumor stage and patient survival. Compared with A3O cells, Trop2 overexpressed in A1780 and SKOV3 cells ( P<0.05). The relative expression levels of Trop2 mRNA in SKOV3-NC group and SKOV3-shRNA group were 1.18±0.24 and 0.42±0.08, with statistically significant difference ( P<0.05). The results of CCK-8 array showed that the cell viability of SKOV3-NC group was significantly higher than that of SKOV3-shRNA group ( P<0.05). The proportion of G 0/G 1 cells in SKOV3-NC and SKOV3-shRNA groups were (38.67±4.22)% and (60.24±8.17)%, respectively. G 0/G 1 arrest was observed in SKOV3-shRNA cells ( P<0.05). The apoptosis rate of SKOV3-shRNA group was (26.32±1.81)%, significantly higher than (6.54±1.32)% of SKOV3-NC group ( P<0.05). The number of migrating SKOV3 cells in the SKOV3-shRNA and SkOV3-NC groups were 1 255.83±108.44 and 1 679.71±213.92, while the number of invading cells were 242.49±52.09 and 473.54±73.11, respectively. Compared with the SKOV3-NC group, the number of migrating and invading SKOV3-shRNA group was significantly reduced (all P<0.05). The expressions of p-AKT2, Bcl-2, vimentin and β-catenin were down-regulated, and the expressions of caspase 3 and E-cadherin were up-regulated in SKOV3-shRNA cells. There was no significant change in the total protein level of AKT. Conclusions:Trop2 expression is related to ovarian cancer stage and postoperative survival. Trop2 can promote ovarian cancer cell proliferation and metastasis by activating the AKT/β-catenin signaling pathway and knockdown of Trop2 inhibits the progression of ovarian cancer.

Objective:To explore the role and molecular mechanism of trophoblastic cell surface antigen 2 (Trop2) in the invasion and migration of ovarian cancer.Methods:Through the data mining of Cancer Cell Line Encyclopedia and TCGA database, the clinical significance of Trop2 expression was analyzed. Western blot was used to detect Trop2 protein expression in ovarian cancer cell lines including A3O, A1780 and SKOV3. SKOV3 cells were used to construct Trop2-short hairpin RNA (shRNA) cell model. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the SKOV3 mRNA expression in SKOV3-shRNA and SKOV3-NC cells. Cell counting kit-8 (CCK8) was used to detect the proliferation of SKOV3-shRNA cells and SKOV3-NC cells. Flow cytometry was used to detect cell cycle and apoptosis in two groups of cells. Transwell array was used to detecte the invasion and migration of SKOV3-shRNA cells and SKOV3-NC cells. Western blot was used to detect the protein expressions of AKT, p-AKT, β-catenin, caspase3, bcl-2, E-cadherin and vimentin.Results:Trop2 mRNA highly expressed in ovarian cancer, and was related to the tumor stage and patient survival. Compared with A3O cells, Trop2 overexpressed in A1780 and SKOV3 cells ( P<0.05). The relative expression levels of Trop2 mRNA in SKOV3-NC group and SKOV3-shRNA group were 1.18±0.24 and 0.42±0.08, with statistically significant difference ( P<0.05). The results of CCK-8 array showed that the cell viability of SKOV3-NC group was significantly higher than that of SKOV3-shRNA group ( P<0.05). The proportion of G 0/G 1 cells in SKOV3-NC and SKOV3-shRNA groups were (38.67±4.22)% and (60.24±8.17)%, respectively. G 0/G 1 arrest was observed in SKOV3-shRNA cells ( P<0.05). The apoptosis rate of SKOV3-shRNA group was (26.32±1.81)%, significantly higher than (6.54±1.32)% of SKOV3-NC group ( P<0.05). The number of migrating SKOV3 cells in the SKOV3-shRNA and SkOV3-NC groups were 1 255.83±108.44 and 1 679.71±213.92, while the number of invading cells were 242.49±52.09 and 473.54±73.11, respectively. Compared with the SKOV3-NC group, the number of migrating and invading SKOV3-shRNA group was significantly reduced (all P<0.05). The expressions of p-AKT2, Bcl-2, vimentin and β-catenin were down-regulated, and the expressions of caspase 3 and E-cadherin were up-regulated in SKOV3-shRNA cells. There was no significant change in the total protein level of AKT. Conclusions:Trop2 expression is related to ovarian cancer stage and postoperative survival. Trop2 can promote ovarian cancer cell proliferation and metastasis by activating the AKT/β-catenin signaling pathway and knockdown of Trop2 inhibits the progression of ovarian cancer.

Objective:To explore the effects of acute paraquat poisoning on cognitive function of patients through neuropsychologic test.Methods:In June 2019, 36 patients with acute paraquat poisoning in the emergency department of a provincial hospital in Hebei Province were selected as the case group. 36 healthy individuals were selected as control group. The cognitive function and depressive state were assessed by mini mental state scale, auditory word learning test, digit span test, connection test, Boston Naming Test and geriatric depression scale.Results:The results of Mini-Mental State examination showed that the total score of the case group was lower than that of the control group, the difference was statistically significant ( P<0.05) . The results of the Auditory Vocabulary Learning test showed that the scores of delayed recall, clue recall, corrective ability and semantic learning strategies of the case group were significantly lower than those in the control group ( P<0.05) . There was no significant difference in the scores of immediate memory between the two groups ( P>0.05) . The scores of Digit Span test and Boston Naming test in the control group were higher than those in the case group, the Trail Making test time in the control group was shorter than that in the case group, and the differences were statistically significant ( P<0.05) . Conclusion:Acute paraquat poisoning can impair human cognitive ability to a certain extent.

Objective:To explore the effects of acute paraquat poisoning on cognitive function of patients through neuropsychologic test.Methods:In June 2019, 36 patients with acute paraquat poisoning in the emergency department of a provincial hospital in Hebei Province were selected as the case group. 36 healthy individuals were selected as control group. The cognitive function and depressive state were assessed by mini mental state scale, auditory word learning test, digit span test, connection test, Boston Naming Test and geriatric depression scale.Results:The results of Mini-Mental State examination showed that the total score of the case group was lower than that of the control group, the difference was statistically significant ( P<0.05) . The results of the Auditory Vocabulary Learning test showed that the scores of delayed recall, clue recall, corrective ability and semantic learning strategies of the case group were significantly lower than those in the control group ( P<0.05) . There was no significant difference in the scores of immediate memory between the two groups ( P>0.05) . The scores of Digit Span test and Boston Naming test in the control group were higher than those in the case group, the Trail Making test time in the control group was shorter than that in the case group, and the differences were statistically significant ( P<0.05) . Conclusion:Acute paraquat poisoning can impair human cognitive ability to a certain extent.

Objective To investigate the expression and role of regulatory plasma cells in gravidas with systemic lupus erythematosus ( SLE) . Methods Gravidas with SLE were enrolled in Henan Provincial People's Hospital from April 2013 to April 2018. They were divided into three groups including pregnancy control group, SLE stable group and SLE deterioration group. The ratio of CD3-LAG-3+CD138high regulatory plasma cells was detected by flow cytometry. The concentrations of soluble human leukocyte antigen-G ( sHLA-G) and anti-nuclear antibody Ig were detected by ELISA. Lymphocytes in peripheral blood of SLE deterioration group were isolated, and then cultured in RPMI1640 medium containing 10％ fetal bovine ser-um and stimulated with HLA-G. Results Flow cytometry showed that the proportion of regulatory plasma cells in SLE stable group was (2. 483±0. 1318)％ and that in SLE deteriorating group was (1. 662± 0. 1304)％. There was a significant difference between the two groups (t=4. 431, P=0. 0013). The con-centrations of sHLA-G in SLE stable group and SLE deteriorating group were (36. 50±3. 510) ng/ml and (16. 50±2. 405) ng/ml, and the difference between the two groups was statistically significant (t=4. 701, P=0. 0008). Correlation analysis showed that the concentration of sHLA-G was positively correlated with the proportion of regulatory plasma cells (r=0. 7471, P=0. 0009). The results of in vitro experiment showed that the proportions of B cells and regulatory plasma cells were ( 7. 573 ± 0. 6539 )％ and ( 1. 593 ± 0.1879)％ in SLE deterioration group and (3. 732±0. 7178)％ and (2. 503±0. 2921)％ in HLA-G group with statistical differences between the two groups (t=3. 957, P=0. 0027;t=2. 620, P=0. 0256). Conclusions The proportion of regulatory plasma cells and the concentration of sHLA-G were significantly decreased in pregnant patients with SLE, which was closely related to disease severity. HLA-G played an important role in promoting the proliferation of regulatory plasma cells.