The present study employed UPLC-MS/MS to analyze and identify compounds in the raw powder of Tongluo Shenggu capsules. An HPLC method for the determination of vitexin content was established. The analysis of this drug was performed on a 30 ℃ thermostatic Acquity UPLC® BEH C18 (2.1 mm×100 mm,1.7 μm) column, with the mobile phase comprising 0.2% formic acid-methanol flowing at 0.3 mL /min in a gradient elution manner. Mass spectrometry was detected by ESI sources in both positive and negative ion modes for qualitative identification of chemical constituents. 12 flavonoid and 3 stilbenes compounds in the raw powder of Tongluo Shenggu capsules were successfully identified. Additionally, an HPLC method for the determination of vitexin content was established using a XBridge C18 column (4.6 mm × 250 mm, 5 µm) with a mobile phase of 0.05% glacial acetic acid in methanol for gradient elution, at a column temperature of 30 °C, a flow rate of 1.0 mL/min, and an injection volume of 20 μL. The method demonstrated good linearity in the concentration range of 10 µg/mL to 40 µg/mL (R=1.000) with an average recovery rate of 96.7%. The establishment of these methods provides a scientific basis for the quality control and development of the raw powder of Tongluo Shenggu capsules.

In recent years, China has been facing the dual challenges of declining fertility rates and births, with male reproductive health issues, especially the decline in semen quality, identified as a pivotal contributor to this phenomenon. Meanwhile, accumulating evidence indicates that air pollutants, an increasingly severe environmental problem, can damage semen quality not only directly through their biological toxicity but also indirectly by disrupting the composition of microbial communities in the gut and semen, thereby dysregulating immune function, endocrine homeostasis, and oxidative stress responses. The gut microbiota and semen microbiota, as important components of the human microecosystem, play crucial roles in maintaining reproductive health. This article comprehensively reviewed the research progress on the potential effects of air pollutants (particulate matter and gaseous pollutants), gut microbiota, and semen microbiota on semen quality. Specifically, it elucidated the mechanisms of interaction between these factors and explored how they affect male fertility.

Normal heart function depends on complex regulation by the brain, and abnormalities in the brain‒heart axis affect various diseases, such as myocardial infarction and anxiety disorders. However, systematic tracking of the brain regions associated with the input and output of the heart is lacking. In this study, we injected retrograde transsynaptic pseudorabies virus (PRV) and anterograde transsynaptic herpes simplex virus (HSV) into the left ventricular wall of mice to identify the whole-brain regions associated with the input to and output from the heart. We successfully detected PRV and HSV expression in at least 170 brain subregions in both male and female mice. Sex differences were discovered mainly in the hypothalamus and medulla, with male mice exhibiting greater correlation and hierarchical clustering than female mice, indicating reduced similarity and increased modularity of virus expression patterns in male mice. Further graph theory and multiple linear regression analysis of different injection timelines revealed that hub regions of PRV had highly similar clusters, with different brain levels, suggesting a top-down, hierarchically transmitted neural control pattern of the heart. Hub regions of HSV had scattered clusters, with brain regions gathered in the cortex and brainstem, suggesting a bottom-up, leapfrog, multipoint neural sensing pattern of the heart. Both patterns contain many hub brain regions that have been previously overlooked in brain‒heart axis studies. These results provide brain targets for future research and will lead to deeper insight into the brain mechanisms involved in specific heart conditions.
Animals ; Male ; Female ; Heart/physiology* ; Mice ; Herpesvirus 1, Suid ; Brain/physiology* ; Mice, Inbred C57BL ; Brain Mapping ; Efferent Pathways/physiology* ; Afferent Pathways/physiology* ; Simplexvirus ; Sex Characteristics

Objective In this study,the gas exchange performance and impact patterns of microalgae culture based on hollow fiber membrane technology were investigated for the culture of space microalgae.Methods The effects of parameters and conditions such as gas flow rate,membrane area and liquid flow rate on the CO2 fixation efficiency and fixation rate of hollow fiber membrane modules were studied,and the gas exchange performance of different parameters were compared by fitting formula.Results The gas flow rate and membrane area have a significant effect on the gas exchange performance of the module.With the increase of gas flow rate,the fixation rate of CO2 increases at first and then stabilizes,and the fixation efficiency of CO2 shows a continuous downward trend;with the increase of membrane area,the fixation rate and fixation efficiency of CO2 increase significantly,while the liquid flow rate has no significant effect on the gas exchange performance of the module.The highest CO2 fixation rate was 168.24 mg/(L·h)when the membrane area was 0.3m2 and the gas flow rate was 2.0 L/min.Conclusion The use of hollow fiber membrane technology can solve the problem of two-phase flow management in the gas exchange between algae culture and atmosphere,which has a better effect on gas exchange,and can provide reference for the design of gas exchange module of space microalgae photobioreactor.

Objective:To investigate the effects of Porphyromonas gingivalis derived outer membrane vesicles (Pg OMV) on osteoclast differentiation of macrophages and its underlying mechanisms. Methods:The morphology and the size distribution of Pg OMV were analyzed by transmission electron microscopy and nanoparticle tracing analysis, respectively. The osteoclast precursors were treated with 1, 3 and 10 mg/L Pg OMV (1, 3 and 10 mg/L OMV treatment group) or phosphate buffer solution (PBS)(control group). The formation of osteoclasts was analyzed by tartrate-resistant acid phosphase (TRAP) staining and F-actin staining and real-time quantitative PCR (RT-qPCR) were used to detect the expression of Fos and matrix metallopeptidase 9 (MMP9). Polymyxin B (PMB) was used to block lipopolysaccharide (LPS) and then Pg OMV was used to treat osteoclast precursor (PMB-OMV treatment group), and OMV treatment group was used as control. TRAP and F-actin staining were used to observe the formation of osteoclasts and actin rings. The effect of Pg OMV on the expression of Toll-like receptor (TLR) 2 and TLR4 in preosteoclasts was detected by Western blotting. The osteoclast precursors were pretreated with 10, 50, 100 and 200 μmol/L C29, an inhibitor of TLR2, and then treated with Pg OMV(OMV+10, 50, 100 and 200 μmol/L C29 treatment group) and OMV treatment group without C29 pretreatment was control. TRAP and F-actin staining were used to observe the formation of osteoclasts and actin rings. The osteoclast precursor cells were treated with OMV (OMV treatment group) and OMV incubated with PMB (PMB-OMV treatment group) and the expression of TLR2 in osteoclast precursor was detected by Western blotting.Results:Pg OMV showed classical vesicular structures, and the average particle size of Pg OMV were 179.2 nm. A large number of actin rings were observed in the 3 and 10 mg/L OMV treatment groups. The percentages of TRAP-positive osteoclast area in 3 mg/L OMV treatment group [(22.6±2.1)%] and 10 mg/L OMV treatment group [(32.0±2.3)%] were significantly increased compared with control group [(4.9±0.5)%] ( P<0.001). Compared with the control group (1.000±0.029), the mRNA relative expression of Fos in 3 mg/L OMV treatment group (1.491±0.114) and 10 mg/L OMV treatment group (1.726±0.254) was significantly increased ( P=0.013, P=0.001). Compared with the control group (1.007±0.148), the mRNA relative expression of MMP9 in the group of 10 mg/L OMV (2.232±0.097) was significantly increased ( P<0.001). Actin ring formation was less in PMB-OMV treatment groups than in OMV treatment groups. The proportion of TRAP-positive osteoclasts area [(14.8±3.8)%] in PMB-OMV treatment group was significantly lower than OMV treatment group [(31.5±6.7) %] ( P=0.004). The relative expression of TLR2 in OMV treatment group (1.359±0.134) was significantly higher than that in the control group (1.000±0.000) ( t=4.62, P=0.044). Compared with the OMV treatment group [(29.4±1.7)%], 50, 100 and 200 μmol/L C29 significantly decreased the formation of osteoclasts [(24.0±1.7)%, (18.5±2.1)%, (9.1±1.3) %] ( P=0.026, P<0.001, P<0.001). TLR2 protein expression in PMB-OMV group (0.780±0.046) was significantly lower than that in OMV group (1.000±0.000)( t=8.32, P=0.001). Conclusions:Pg OMV can promote osteoclast differentiation by carrying LPS, TLR2 plays an important role in Pg OMV mediated osteoclast differentiation.

BACKGROUND:With computer-aided design,melt electrowriting technology can precisely construct 3D tissue engineering scaffolds with specific morphology,which has attracted increasing attention in tissue engineering. OBJECTIVE:To elaborate on the progress of melt electrowriting technology in tissue engineering in recent years. METHODS:PubMed and CNKI were used to retrieve articles about applications of melt electrowriting technology in tissue engineering.The search time was from March 2008 to February 2023.The search terms were"melt electrowriting,melt electrospinning,electrospinning,tissue engineering,scaffold,regeneration"in English and"melt electrowriting,electrospinning,tissue engineering"in Chinese.A preliminary screening of articles was performed by reading the titles and abstracts.Finally,69 articles were included for review. RESULTS AND CONCLUSION:(1)Melt electrowriting technology can achieve precise layer-by-layer deposition of fibers compared to traditional electrospinning technology,which better simulates the complex structure of natural tissues.Compared to other 3D printing technologies,smaller-diameter fibers can be prepared by melt electrowriting technology,resulting in highly ordered porous structures.(2)By combining with other scaffold preparation techniques or materials,such as fused deposition modeling,solution electrospinning technology,and hydrogel,melt electrowriting technology shows great potential in preparing complex tissue engineering scaffolds,which provides certain possibilities for achieving complex tissue regeneration.(3)The regeneration of complex tissues often involves blood vessels,nerves,and soft and hard tissues at the same time.The regeneration of blood vessels and nerves is of great significance to realize the physiological reconstruction of tissues.However,soft and hard tissues have certain difficulties to realize the coordinated regeneration of both due to their different biological and mechanical properties.Melt electrowriting technology has certain advantages in the field of bionic scaffolds due to its good biocompatibility,the ability to prepare multi-scale scaffolds and high porosity.

Objective : To investigate the impact of dexmedetomidine on the oncological behavior of hepatocellular carcinoma and explore the role of NF-E2-related factor 2 (Nrf2) at both in vitro and in vivo levels． Methods : In vivo experiment，Male C57BL/6J mice were randomly divided into a control group ( Ctrl group) ，a hepatocellular carcinoma group ( HCC group) ，and a hepatocellular carcinoma + dexmedetomidine group ( HCC + Dex group) . Hepatocellular carcinoma was induced in mice by combining N-Nitrosodiethylamine ( DEN) / carbon tetrachloride ( CCl4 ) ，followed by daily intraperitoneal injection of 10% dexmedetomidine for two weeks．After feeding the mice for one month，the mice were assessed for the quantity and size of liver tumors．The proliferation ability of liver cancer was evaluated using Ki67 immunohistochemistry．Additionally，the expression level of Nrf2 protein in tumor tissue was measured through immunofluorescence．In vitro experiment，Hepa1-6 cells were incubated with different concentrations of dexmedetomidine (0. 1，1，5 nmol /L) for 48 hours to examine their effects．The proliferation， migration and invasion abilities of Hepa1-6 cells were evaluated using the MTT and Transwell methods．The expres- sion level of Nrf2 protein in the Hepa1-6 cells was measured using Western blot and immunofluorescence．Addition- ally，the proliferation ，migration and invasion abilities of cells were assessed after Nrf2 knockdown via si-RNA transfection，in combination with incubation with 1 nmol /L dexmedetomidine for 48 hours. Results : ompared to the HCC group，the anatomical examination results revealed an increase in the number of liver tumors and the lon- gest diameter in the HCC + Dex group (P ＜0. 05) ． Ki67 immunohistochemistry results indicated the number of Ki67 positive cells in liver cancer tissue increased in the HCC + Dex group (P＜0. 01) ．The immunofluorescence assay demonstrated an upregulation of Nrf2 expression level in the HCC + Dex group (P ＜0. 05 ) ． MTT results showed that 1 nmol /L of dexmedetomidine increased the cell viability of Hepa1-6 cells (P＜0. 05) ．Transwell re- sults indicated that 0. 1 ，1 ，and 5 nmol /L of dexmedetomidine enhanced the invasive ability of Hepa1-6 cells， while 0. 1 and 1 nmol /L of dexmedetomidine enhanced the migration ability (P＜0. 05) ．Western blot and immu- nofluorescence results showed an upregulation of Nrf2 expression level in cells after treatment with 1 nmol /L dexme- detomidine (P＜0. 01) ．The Nrf2 expression level of cells was reduced using si-RNA，followed by treatment with 1 nmol /L dexmedetomidine．The results from MTT and Transwell assays revealed a decrease in the viability，invasion and migration ability of Hepa1-6 cells (P＜0. 01) ． Conclusion Dexmedetomidine may enhance the proliferation， invasion and migration capacity of hepatocellular carcinoma by upregulating the expression of Nrf2 .

Objective In order to use plants to absorb high concentration carbon dioxide in closed space,the gas exchange capacity and regulation measures of six horticultural plants were explored.Methods The gas exchange characteristics of Bamboo Palm,Chrysanthemum,Peace Lily,Anthurium,Aloe vera,and Nephrolepis exaltata were studied under three light gradients[170 μmol/(m2·s),270 μmol/(m2·s),370 μmol/(m2·s)]and four carbon dioxide concentration gradients(500 ppm,1 000 ppm,1 500 ppm,2 000 ppm)using a closed low-pressure device.Results The results indicate significant differences in the average photosynthetic rates among the six plants,with the ranking order as Chrysanthemum>Peace Lily>Bamboo Palm>Aloe vera>Anthurium>Nephrolepis exaltata.In contrast,the transpiration rate and respiration rate of Chrysanthemum,Peace Lily and Bamoo Palm were higher,aloe vera absorbed carbon dioxide under dark conditions.Under a light intensity of 370 μmol/(m2·s),the photosynthetic rate of Bamboo Palm was most enhanced,2.57 times that under the condition of 170 μmol/(m2·s),whereas high light intensity suppressed the photosynthetic rate of Nephrolepis exaltata,which was about 3%lower than that under low light intensity.Under a carbon dioxide concentration of 2 000 ppm,the photosynthetic rates of Chrysanthemum,Peace Lily,and Bamboo Palm was more than 9.13 μmol CO2/(m2·s),approximately three times the average rate under low carbon dioxide concentration.Conclusion Among the six horticultural plants,Chrysanthemum,Bamboo Palm,and Peace Lily exhibit superior gas exchange capabilities,and increasing the light intensity and carbon dioxide concentration in the cultivation environment significantly enhances the photosynthetic rate of these three plants.

Coding with high-frequency stimuli could alleviate the visual fatigue of users generated by the brain-computer interface (BCI) based on steady-state visual evoked potential (SSVEP). It would improve the comfort and safety of the system and has promising applications. However, most of the current advanced SSVEP decoding algorithms were compared and verified on low-frequency SSVEP datasets, and their recognition performance on high-frequency SSVEPs was still unknown. To address the aforementioned issue, electroencephalogram (EEG) data from 20 subjects were collected utilizing a high-frequency SSVEP paradigm. Then, the state-of-the-art SSVEP algorithms were compared, including 2 canonical correlation analysis algorithms, 3 task-related component analysis algorithms, and 1 task discriminant component analysis algorithm. The results indicated that they all could effectively decode high-frequency SSVEPs. Besides, there were differences in the classification performance and algorithms' speed under different conditions. This paper provides a basis for the selection of algorithms for high-frequency SSVEP-BCI, demonstrating its potential utility in developing user-friendly BCI.
Humans ; Brain-Computer Interfaces ; Evoked Potentials, Visual ; Algorithms ; Discriminant Analysis ; Electroencephalography

Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.