ObjectiveTo investigate the mechanism of action of Kaixinsan in the treatment of Alzheimer's disease （AD） based on network pharmacology， molecular docking， and animal experimental validation. MethodsThe Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） and the Encyclopedia of Traditional Chinese Medicine（ETCM） databases were used to obtain the active ingredients and targets of Kaixinsan. GeneCards， Online Mendelian Inheritance in Man（OMIM）， TTD， PharmGKB， and DrugBank databases were used to obtain the relevant targets of AD. The intersection （common targets） of the active ingredient targets of Kaixinsan and the relevant targets of AD was taken， and the network interaction analysis of the common targets was carried out in the STRING database to construct a protein-protein interaction（PPI） network. The CytoNCA plugin within Cytoscape was used to screen out the core targets， and the Metascape platform was used to perform gene ontology（GO） functional enrichment analysis and Kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment analysis. The “drug-active ingredient-target” interaction network was constructed with the help of Cytoscape 3.8.2， and AutoDock Vina was used for molecular docking. Scopolamine （SCOP） was utilized for modeling and injected intraperitoneally once daily. Thirty-two male C57/BL6 mice were randomly divided into blank control （CON） group （0.9% NaCl， n=8）， model （SCOP） group （3 mg·kg^-1·d^-1， n=8）， positive control group （3 mg·kg^-1·d^-1 of SCOP+3 mg·kg^-1·d^-1 of Donepezil， n=8）， and Kaixinsan group （3 mg·kg^-1·d^-1 of SCOP+6.5 g·kg^-1·d^-1 of Kaixinsan， n=8）. Mice in each group were administered with 0.9% NaCl， Kaixinsan， or Donepezil by gavage twice a day for 14 days. Morris water maze experiment was used to observe the learning memory ability of mice. Hematoxylin-eosin （HE） staining method was used to observe the pathological changes in the CA1 area of the mouse hippocampus. Enzyme linked immunosorbent assay（ELISA） was used to determine the serum acetylcholine （ACh） and acetylcholinesterase （AChE） contents of mice. Western blot method was used to detect the protein expression levels of signal transducer and activator of transcription 3（STAT3） and nuclear transcription factor（NF）-κB p65 in the hippocampus of mice. ResultsA total of 73 active ingredients of Kaixinsan were obtained， and 578 potential targets （common targets） of Kaixinsan for the treatment of AD were screened out. Key active ingredients included kaempferol， gijugliflozin， etc.. Potential core targets were STAT3， NF-κB p65， et al. GO functional enrichment analysis obtained 3 124 biological functions， 254 cellular building blocks， and 461 molecular functions. KEGG pathway enrichment obtained 248 pathways， mainly involving cancer-related pathways， TRP pathway， cyclic adenosine monophosphate（cAMP） pathway， and NF-κB pathway. Molecular docking showed that the binding of the key active ingredients to the target targets was more stable. Morris water maze experiment indicated that Kaixinsan could improve the learning memory ability of SCOP-induced mice. HE staining and ELISA results showed that Kaixinsan had an ameliorating effect on central nerve injury in mice. Western blot test indicated that Kaixinsan had a down-regulating effect on the levels of NF-κB p65 phosphorylation and STAT3 phosphorylation in the hippocampal tissue of mice in the SCOP model. ConclusionKaixinsan can improve the cognitive impairment function in SCOP model mice and may reduce hippocampal neuronal damage and thus play a therapeutic role in the treatment of AD by regulating NF-κB p65， STAT3， and other targets involved in the NF-κB signaling pathway.

The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas (CRISPR-associated proteins) is an adaptive immune system present in most bacteria and archaea, protecting them from infection by exogenous genetic elements. Due to its simplicity, cost-effectiveness, and precise gene editing capabilities, CRISPR/Cas technology has emerged as a promising tool for treating diseases. The continuous refinement of derivative systems has further broadened its scope in disease treatment. Nevertheless, the heterogeneous physiopathological nature of diseases and variations in disease onset sites pose significant challenges for in vivo applications of CRISPR systems. The efficiency of CRISPR systems in disease treatment is directly influenced by the performance of the delivery system. Additionally, concerns such as off-target effects present crucial hurdles in the clinical implementation of CRISPR systems. This review provides a comprehensive overview of the development of CRISPR systems, vector technologies, and their applications in disease treatment, while also addressing the challenges encountered in clinical settings. Furthermore, future research directions are outlined to pave the way for advancements in CRISPR-based therapies.

CRISPR/Cas9-based therapeutics face significant challenges in penetrating the dense microenvironment of solid tumors, resulting in insufficient gene editing and compromised treatment efficacy. Current nanostrategies, which mainly focus on the paracellular pathway attempted to improve gene editing performance, whereas their efficiency remains uneven in the heterogenous extracellular matrix. Here, the nanoCRISPR system is prepared with self-cascading mechanisms for gene editing-mediated robust apoptosis and transcellular penetration. NanoCRISPR unlocks its self-cascade capability within the matrix metallopeptidase 2-enriched tumor microenvironment, initiating the transcellular penetration. By facilitating cellular uptake, nanoCRISPR triggers robust apoptosis in edited malignancies, promoting further transcellular penetration and amplifying gene editing in neighboring tumor cells. Benefiting from self-cascade between robust apoptosis and transcellular penetration, nanoCRISPR demonstrates continuous gene transfection/tumor killing performance (transfection/apoptosis efficiency: 1st round: 85%/84.2%; 2nd round: 48%/27%) and homogeneous penetration. In xenograft tumor-bearing mice, nanoCRISPR treatment achieves remarkable anti-tumor efficacy (∼83%) and significant survival benefits with minimal toxicity. This strategy presents a promising paradigm emphasizing transcellular penetration to enhance the effectiveness of CRISPR-based antitumor therapeutics.

Background::Carotid intima-media thickness (IMT) and diameter, stiffness, and wave reflections, are independent and important clinical biomarkers and risk predictors for cardiovascular diseases. The purpose of the present study was to establish nationwide reference values of carotid properties for healthy Chinese adults and to explore potential clinical determinants.Methods::A total of 3053 healthy Han Chinese adults (1922 women) aged 18-79 years were enrolled at 28 collaborating tertiary centers throughout China between April 2021 and July 2022. The real-time tracking of common carotid artery walls was achieved by the radio frequency (RF) ultrasound system. The IMT, diameter, compliance coefficient, β stiffness, local pulse wave velocity (PWV), local systolic blood pressure, augmented pressure (AP), and augmentation index (AIx) were then automatically measured and reported. Data were stratified by age groups and sex. The relationships between age and carotid property parameters were analyzed by Jonckheere-Terpstra test and simple linear regressions. The major clinical determinants of carotid properties were identified by Pearson’s correlation, multiple linear regression, and analyses of covariance.Results::All the parameters of carotid properties demonstrated significantly age-related trajectories. Women showed thinner IMT, smaller carotid diameter, larger AP, and AIx than men. The β stiffness and PWV were significantly higher in men than women before forties, but the differences reversed after that. The increase rate of carotid IMT (5.5 μm/year in women and 5.8 μm/year in men) and diameter (0.03 mm/year in both men and women) were similar between men and women. For the stiffness and wave reflections, women showed significantly larger age-related variations than men as demonstrated by steeper regression slopes (all P for age by sex interaction <0.05). The blood pressures, body mass index (BMI), and triglyceride levels were identified as major clinical determinants of carotid properties with adjustment of age and sex. Conclusions::The age- and sex-specific reference values of carotid properties measured by RF ultrasound for healthy Chinese adults were established. The blood pressures, BMI, and triglyceride levels should be considered for clinical application of corresponding reference values.

Objective:To explore the relationship between preterm labor with different causes and cerebral perfusion in different regions of interest in very preterm infants.Methods:This was a prospective cohort study. A total of 145 preterm infants with gestational age of 28-31 +6 weeks who were hospitalized in the Neonatology Department of the Third Affiliated Hospital of Zhengzhou University within 24 h after birth from April 2022 to May 2023 were selected for the study, and were categorized into the iatrogenic preterm labor group ( n=55), spontaneous preterm labor with premature rupture of the membranes (PROM) group ( n=47), and spontaneous preterm labor with intact membranes group ( n=43) according to the cause of preterm labor. Cerebral blood flow (CBF) values in the cortex and deep gray matter of different regions of interest (frontal lobe, temporal lobe, parietal lobe, occipital lobe, thalamus, and basal ganglia) were measured using the arterial spin labeling technique in the very preterm infants in each group. One-way analysis of variance, Kruskal-Wallis H test and Bonferroni correction, Chi-square test or Fisher's exact probability method, analysis of covariance, and LSD test were used to compare the differences in CBF among the groups. Results:The differences in the incidence of complications such as cerebral white matter injury, Ⅰ-Ⅱ grade intracranial hemorrhage, and late-onset sepsis during hospitalization among the three groups of preterm infants were not statistically significant (all P>0.05). In the iatrogenic preterm labor group, compared with the spontaneous preterm labor with PROM group, CBF [in units of ml/ (100 g·min)] was higher in regions of interest such as the right temporal lobe [20.5 (16.1-24.6) vs. 17.1 (14.5-23.0)], bilateral parietal lobe [left side: 22.4 (17.1-25.3) vs. 16.9 (14.4-24.1); right side: 23.0 (18.2-27.4) vs. 17.0 (14.0-22.2)], right occipital lobe [22.1 (18.6-29.5) vs. 19.4 (13.7-24.5)], bilateral basal ganglia [left side: 33.0 (29.1-36.3) vs. 24.9 (22.9-33.1); right side: 32.8 (29.0-37.0) vs. 26.1 (22.3-35.0)], and bilateral thalamus [left side: 39.2 (36.0-45.0) vs. 32.6 (25.1-42.2); right side: 38.6 (34.6-44.1) vs. 32.0 (25.4-44.9)] (Bonferroni corrected, all P<0.017). Compared with the spontaneous preterm labor group with intact membranes, CBF in the iatrogenic preterm labor group was higher in the cortex and deep gray matter of regions of interest such as bilateral frontal lobe [left side: 21.4 (18.3-25.3) vs. 17.0 (12.0-22.2); right side: 22.1 (16.7-25.0) vs. 15.9 (12.0-23.3)], temporal lobe [left side: 21.4 (17.0-24.8) vs. 18.4 (14.0-22.0); right side: 20.5 (16.1-24.6) vs. 17.3 (13.3-22.3)], parietal lobe [left side: 22.4 (17.1-25.3) vs. 15.3 (10.4-20.8); right side: 23.0 (18.2-27.4) vs. 15.7 (11.1-23.6)], occipital lobe [left side: 22.7 (18.8-28.4) vs. 18.2 (11.4-23.4); right side: 22.1 (18.6-29.5) vs. 19.6 (14.0-25.8)], basal ganglia [left side: 33.0 (29.1-36.3) vs. 27.7 (19.1-32.4); right side: 32.8 (29.0-37.0) vs. 27.7 (21.5-33.0)] and thalamus [left side: 39.2 (36.0-45.0) vs. 33.9 (26.0-43.7); right side: 38.6 (34.6-44.1) vs. 33.3 (27.8-40.4)] (Bonferroni corrected, all P<0.017). Analysis of covariance revealed that the cause of preterm birth had a significant effect on CBF values in the cortex and deep gray matter of very preterm infants ( P=0.007), and that iatrogenic preterm birth elevated CBF perfusion in the localized cerebral cortex and deep gray matter of very preterm infants as compared to the spontaneous preterm births with PROM group and spontaneous preterm births with intact membranes group (LSD test, all P<0.05). Conclusion:Cerebral blood perfusion in very preterm infants is related to the causes leading to preterm birth, and local cortical and deep gray matter blood perfusion levels in the brain are increased in those with iatrogenic preterm birth compared to spontaneous preterm birth.

Objective:To reveal the diurnal variation characteristics of the exercise adaptability of cardiac function in healthy men of different ages by the analysis of echocardiographic parameter changes before and after exercise intervention using the left ventricular pressure-strain loop techniques.Methods:A prospective control design was used to collect 193 healthy male volunteers from October 2022 to July 2023 in Tangdu Hospital, 101 in the young group [(26±5) years old], and 92 in the middle-aged group [(50±4) years old]. The heart rate, blood pressure and routine echocardiographic parameters were collected at rest and after exercise stress (3-minute step test) at 7∶00 a.m.and 10∶00 p.m., respectively. The heart rate, blood pressure cardiac output, and echocardiographic parameters [anteroposterior diameter of left atrium (LAD), ratio of peak early diastolic flow velocity to late diastolic peak flow velocity of mitral valve (E/A), left ventricular ejection fraction (LVEF), left ventricular end diastolic diameter (LVEDD), left ventricular end systolic diameter (LVEDS)] were then determined. The parameters of myocardial work including left ventricular longitudinal strain (GLS), global work index (GWI), global work efficiency (GWE), global constructive work (GCW) and global wasted work (GWW) were obtained by left ventricular pressure-strain loop technique. The differences of the parameters above between before and after step test, morning and night, young group and middle-aged group were compared.Results:The heart rate, blood pressure and cardiac output of young and middle-aged healthy men increased significantly in the morning and night step test (all P<0.05), GLS, GWI, GCW and GWW were significantly higher than those in resting state (all P<0.05), and GWE was significantly lower than that in resting state ( P<0.05). The change rates of heart rate, cardiac output, GLS, GWI and GCW in the early morning were significantly higher than those at night (all P<0.05), while the change rates of blood pressure and GWE in each group were significantly lower than those at night ( P<0.05). The change rates of heart rate, systolic blood pressure, GLS and GWI in the middle-aged group were significantly higher than those in the young group in the morning and night (all P<0.05), while the change rate of diastolic blood pressure at night was significantly higher than that in the young group ( P<0.05), but there was no significant difference in the morning ( P>0.05). Conclusions:The cardiac adaptability of healthy men has obvious circadian rhythm, characterized by being of low in the morning and high in the evening. Age mainly affects the cardiac adaptability in the morning. The response to low-intensity load stimulation increases with aging, indicating the decrease of morning cardiac adaptability.

Objective:To evaluate the performance of the self-developed transcranial ultrasound patch probe as well as its stability and safety in clinical applications.Methods:This study was a cross-sectional study. Fifty healthy subjects in Tangdu Hospital from October to December 2023 were randomly and equally divided into two groups using a simple randomization method. The transcranial Doppler ultrasound (TCD) examinations were performed using the self-developed transcranial ultrasound patch probe and normal TCD probe alternatively in each group from squatting to sitting and then to standing posture. Two experienced sonographers (A, B) each completed one test for the same subject using the above two different probes. The differences of the ultrasound parameters of middle cerebral artery (MCA) blood flow measured by the self-developed transcranial ultrasound patch probe and the normal TCD probe in different postures (squatting, sitting, standing) were compared. At the same time, operator satisfaction with the use of the two probes was assessed in terms of time taken to obtain satisfactory images, operational comfort, and ease of handling. In addition, the subject's comfort during the examination was examined, and the signal acquisition stability of the two probes was comprehensively evaluated.Results:There was no significant difference in detection of various ultrasound parameters of MCA between using the transcranial patch probe and the normal TCD probe (all P>0.05). The overall satisfaction of transcranial ultrasound patch probe and normal TCD probe was quite similar. The transcranial ultrasound patch probe acquired favorable and stable signals. No subjects experienced adverse events/accidents during the examination of the subjects using the transcranial ultrasound patch probe as well as the normal TCD probe.Both probes had good and stable signal acquisition. Conclusions:TCD examinations could be well performed using the transcranial ultrasound patch probe, which might provide a potential new method for real-time monitoring and evaluating of cerebral blood flow of a moving subject.

Objective:To study the influence of periventricular-intraventricular hemorrhage (PVH-IVH) on cerebral blood flow (CBF) of preterm infants in the late postnatal period using arterial spin labeling (ASL) magnetic resonance imaging (MRI).Methods:From January 2023 to June 2023, 65 preterm infants (gestational age <32 weeks, birth weight <1 500 g) who were born in the Department of Obstetrics, Third Affiliated Hospital of Zhengzhou University and transferred to the Neonatal Intensive Care Unit were included in the prospective study.They were examined by the brain MRI and ASL at the corrected gestational age of 35-40 weeks.According to the results of the brain ultrasound within 1 week after birth, they were divided into the mild IVH group (25 cases) and the non-IVH group (40 cases). The CBF values in regions of interest (frontal lobe, temporal lobe, parietal lobe, occipital lobe, thalamus, and basal ganglia) on ASL images were compared.Multiple linear regression analysis was used to analyze the effect of PVH-IVH on CBF values in different ASL regions of interest, including frontal cortex, temporal cortex, parietal cortex, occipital cortex, thalamus, and basal ganglia.Results:Compared with those of non-IVH group, infants in the mild IVH group presented significantly older gestational age [29.0 (28.5, 30.4) weeks vs.28.2 (27.0, 31.0) weeks, Z=-2.398, P=0.016], higher hematocrit (HCT) in the latest examination prior to the brain MRI [29.6(26.4, 32.3)% vs.27.8 (25.6, 30.5)%, Z=-2.155, P=0.031], and larger body weight at the time of examination [2 015.0 (1 930.0, 2 127.5) g vs.1 950.0 (1 900.0, 1 997.5) g, Z=-3.314, P=0.001]. After adjustment for confounding factors of gestational age at birth, latest HCT and weight at the time of examination, the multivariable linear regression analysis showed that CBF values in the frontal lobe (95% CI: -8.367--4.042; P<0.001), temporal lobe (95% CI: -19.077--2.854; P=0.008), parietal lobe (95% CI: -8.344--3.502; P<0.001), occipital lobe (95% CI: -9.446--3.645; P<0.001), basal ganglia (95% CI: -7.543--1.963; P=0.001) and thalamus (95% CI: -8.051--2.372; P<0.001) were significantly lower in infants of the mild IVH group than those of non-IVH group. Conclusions:At the same corrected gestational age, mild IVH is correlated with low CBF values in local cerebral cortex and subcortical gray matter in premature infants.However, the predictive potential of CBF values in long-term neurological prognosis requires further explorations.

The aim of this paper was to observe the combination therapy with total triterpenoids of Chaenomeles speciosa and omeprazole on indomethacin-induced gastric ulcer in rats, and explore its possible mechanism. Rats were randomly divided into normal group, model group, omeprazole monotherapy(3.6 mg·kg~(-1)) group, total triterpenoids of C. speciosa monotherapy(100 mg·kg~(-1)) group, total triterpenoids of C. speciosa and omeprazole combination therapy(100 mg·kg~(-1)+3.6 mg·kg~(-1)) group. Except for the normal group, the other groups were given indomethacin(20 mg·kg~(-1)) by oral once a day for 7 consecutive days. Then the treated groups were given corresponding drugs by gavage, once a day for 14 consecutive days. The next day after the last administration, half of the rats in each group were measured the gastric mucosal blood flow, gastric juice volume and serum TNF-α, IL-1β, IL-6, IL-4 and IL-10. After the remaining rats in each group were underwent pyloric ligation 4 hours after the last administration, the gastric endocrine volume, pH value and total acidity of gastric secretion were measured, then histological analysis was performed, MPO activity, cAMP content and histomorphological analysis were conducted. Real-time PCR was applied to detect the mRNA expressions of gastric tissue TNF-α,IL-1β, IL-6, IL-4, IL-10, VEGFA, A_(2A)R; the protein expressions of VEGFA, A_(2A)R, PKA, p-PKA, CREB, p-CREB, EGF, EGFR, p-EGFR, MUC6, TFF2 in gastric tissue were detected by Western blot. The results indicated that total triterpenoids of C. speciosa and omeprazole combination therapy might significantly increase gastric mucosal blood flow, gastric mucus volume, reduce gastric endocrine volume, secretion acidity and mucosal damage, decrease the levels of TNF-α,IL-1β and IL-6, increase the levels of IL-4 and IL-10 in blood and gastric tissue, inhibit the activity of MPO, increase the content of cAMP in gastric tissue, up-regulate the mRNA expressions of VEGFA, A_(2A)R and protein expressions of VEGFA, A_(2A)R, PKA, p-PKA, CREB, p-CREB, EGF, EGFR, p-EGFR, MUC6, TFF2 in gastric tissue, elevate p-PKA/PKA, p-CREB/CREB and p-EFGR/EFGR. Moreover, the combination therapy with total triterpenoids of C. speciosa and omeprazole was more obvious than those of two monotherapies. These aforementioned findings suggested that the combination therapy with total triterpenoids of C. speciosa and omeprazole on indomethacin-induced gastric ulcer have significant therapeutic effect on indomethacin induced gastric ulcer in rats, its mechanism might be related to regulating A_(2A)R/AKT/CREB, A_(2A)R/VEGFA, EGF/EGFR and MUC6/TFF2 signaling pathways, inhibiting pro-inflammatory factors, increasing gastric mucosal blood flow, up-regulating mucosal cell proliferation factors and promoting mucosal protective factors.
Animals ; Cytokines ; Gastric Mucosa ; Indomethacin ; Omeprazole ; pharmacology ; Phytochemicals ; pharmacology ; Random Allocation ; Rats ; Rosaceae ; chemistry ; Stomach Ulcer ; chemically induced ; drug therapy ; Triterpenes ; pharmacology ; Tumor Necrosis Factor-alpha

Objective To observe the clinical effect of platelet-rich fibrin (PRF) on prevention of postoperative hemorrhage and facial traumatic swelling in patients with mandibular angle osteotomy.Methods Twenty-five patients with mandibular angle hypertrophy were included in this study from January 2014 to November 2015.Split face comparative study was carried out to use the left and right sides as the experimental side and the control side,respectively.The PRF in the experimental side was placed in the mandibular osteotomy,while the control side was placed in platelet-poor plasma (PPP).After operation,the drainage volume and facial swelling degree were measured.Results The drainage volume of the experimental group (PRF group) was (20.35 ±7.40) ml,the control group (PPP group) was (43.23±11.96) ml,and the difference was statistically significant (P＜0.05).There was no such case without swelling in postoperative third day.The facial swelling score on the experimental side was (1.19±0.40),the control side was (2.62±0.64),and two groups of postoperative facial swelling scores were significantly different (P＜0.05).Conclusions The PRF can reduce postoperative bleeding and facial swelling after mandibular angle osteotomy.