Salidroside Regulates Mitochondrial Homeostasis After Polarization of RAW264.7 Macrophages.

ABSTRACT: Salidroside has anti-inflammatory and antiatherosclerotic effects, and mitochondrial homeostasis imbalance is closely related to cardiovascular disease. The aim of this study was to investigate the effect of salidroside on mitochondrial homeostasis after macrophage polarization and elucidate its possible mechanism against atherosclerosis. RAW264.7 cells were stimulated with 1 µg·mL -1 Lipopolysaccharide and 50 ng·mL -1 IFN-γ establish M1 polarization and were also pretreated with 400 µM salidroside. The relative expression of proinflammatory genes was detected by RT-PCR whereas that of mitochondrial homeostasis-related proteins and nuclear factor kappa-B (NF-κB) was detected by WB. Levels of intracellular reactive oxygen species (ROS), mitochondrial membrane potential, and mass were measured by chemifluorescence whereas that of NF-κB nuclear translocation was detected by immunofluorescence. Compared with the Mφ group, the M1 group demonstrated increased mRNA expression of interleukin-1ß , inductible nitric oxide synthase (iNOS), and tumor necrosis factor-α ; increased protein expression of iNOS, NOD-like receptor protein 3, putative kinase 1 , and NF-κB p65 but decreased protein expression of MFN2, Tom20, and PGC-1α; decreased mitochondrial membrane potential and mass; and increased ROS levels and NF-κB p65 nuclear translocation. Salidroside intervention decreased mRNA expression of interleukin-1ß and tumor necrosis factor-α compared with the M1 group but did not affect that of iNOS. Furthermore, salidroside intervention prevented the changes in protein expression, mitochondrial membrane potential and mass, ROS levels, and NF-κB p65 nuclear translocation observed in the M1 group. In summary, salidroside ultimately inhibits M1 macrophage polarization and maintains mitochondrial homeostasis after macrophage polarization by increasing mitochondrial membrane potential, decreasing ROS levels, inhibiting NF-κB activation, and in turn regulating the expression of proinflammatory factors and mitochondrial homeostasis-associated proteins.

Quantification of Chemical Groups and Quantitative HPLC Fingerprint of Poria cocos (Schw.) Wolf.

(1)Objective: In this study, a quantitative analysis of chemical groups (the triterpenoids, water-soluble polysaccharides, and acidic polysaccharides) and quantitative high liquid performance chromatography (HPLC) fingerprint of Poria cocos (Schw.) Wolf (PC) for quality control was developed. (2) Methodology: First, three main chemical groups, including triterpenoids, water-soluble polysaccharides, and acidic polysaccharides, in 16 batches of PC were evaluated by ultraviolet spectrophotometry. Afterward, the quantitative fingerprint of PC was established, and the alcohol extract of PC was further evaluated. The method involves establishing 16 batches of PC fingerprints by HPLC, evaluating the similarity of different batches of PC, and identifying eight bioactive components, including poricoic acid B (PAB), dehydrotumulosic acid (DTA), poricoic acid A (PAA), polyporenic acid C (PAC), 3-epidehydrotumulosic acid (EA), dehydropachymic acid (DPA), dehydrotrametenolic acid (DTA-1), and dehydroeburicoic acid (DEA), in PC by comparison with the reference substance. Combined with the quantitative analysis of multi-components by a single marker (QAMS), six bioactive ingredients, including PAB, DTA, PAC, EA, DPA, and DEA, in PC from different places were established. In addition, the multivariate statistical analyses, such as principal component analysis and heatmap hierarchical clustering analysis are more intuitive, and the visual analysis strategy was used to evaluate the content of bioactive components in 16 batches of PC. Finally, the analysis strategy of three main chemical groups in PC was combined with the quantitative fingerprint strategy, which reduced the error caused by the single method. (3) Results: The establishment of a method for the quantification of chemical groups and quantitative HPLC fingerprint of PC was achieved as demonstrated through the quantification of six triterpenes in PC by a single marker. (4) Conclusions: Through qualitative and quantitative chemical characterization, a multi-directional, simple and efficient routine evaluation method of PC quality was established. The results reveal that this strategy can provide an analytical method for the quality evaluation of PC and other Chinese medicinal materials.

[Molecular Identification on Three Medicinal Phlomis Species Produced in Yunnan Province Based on ITS and matK].

Objective: To analyze ITS region and matK gene sequences of three medicinal Phlomis plants,in order to provide molecular basis for identifying and protecting their wild resources. Methods: PCR and sequencing were conducted on Phlomis likiangensis,Phlomis melanantha and Phlomis betonicoides wild populations by primers pairs ITS4 / ITS5 and matKXF / matK5 R. Results: The smallest inter-K2 P genetic distance was further than the largest intra-K2 P genetic distance in Phlomis likiangensis, Phlomis melanantha and Phlomis betonicoides. Different samples of three medicinal Phlomis plants were gathered together and could be distinguished from other exogenous species by Neighbor-Joining( NJ) tree. Phlomis likiangensis, Phlomis melanantha and Phlomis betonicoides had three, three and one sites on ITS2 for their effective identification, and had three,three and three sites on ITS1 for their effective identification respectively. Phlomis betonicoides had three sites on matK for its effective identification, while Phlomis likiangensis or Phlomis melanantha needed multiple sites for their effective identification. Conclusion: The study implies that ITS1,ITS2 and matK fragments could be used for molecular identification of Phlomis likiangensis, Phlomis melanantha and Phlomis betonicoides.

Analysis of Factors Influencing Prognosis and Assessment of 60 Cases of Decompensated Cirrhotic Patients with Portal Hypertension.

Objective: To investigate the risk factors for the development of portal hypertension in patients with decompensated cirrhosis and analyze their prognosis. Methods: Patients with decompensated cirrhosis who were admitted to our hospital and Qu fu People's Hospital from June 2022 to June 2023 were included in this study. Among them, there were 45 male and 15 female patients, with a median age of 56 (range: 35-77) years. A comparative analysis was performed between Group A (hepatic venous pressure gradient, HVPG <16 mmHg) and Group B (HVPG ≥16 mmHg) patients, along with various clinical outcomes. Multivariate analysis was conducted to explore the risk factors influencing the occurrence of portal hypertension and adverse prognosis in patients with cirrhosis. Results: In Group A patients with portal hypertension, we observed lower levels of aspartate aminotransferase, laminin, serum hyaluronic acid, type III procollagen N-terminal peptide, total bile acids, and cholylglycine acid compared to Group B. On the other hand, levels of alanine aminotransferase, white blood cells, and serum albumin were higher in Group A than in Group B. These differences between the groups were statistically significant (P < 0.05). Multivariate analysis of the aforementioned risk factors indicated that low white blood cell count, high cholylglycine acid levels, and high serum hyaluronic acid levels were identified as independent risk factors for the occurrence of difficult-to-control complications in decompensated portal hypertension among patients with liver cirrhosis (P < 0.05). Conclusion: Liver cirrhosis patients with portal hypertension and multiple risk factors like low white blood cell count and high liver transaminase levels should be cautious regarding the progression of portal hypertension when combined with splenomegaly, liver fibrosis, and bile stasis, as it often indicates a poor prognosis.

Connexin43 and angiotensin II alterations in hearts of rats having undergone an acute exposure to alcohol.

INTRODUCTION: Alcohol-induced heart damage is associated with enzyme and protein alterations. The purpose of this study was to investigate alcohol-induced alterations in cardiac connexin 43 (Cx43) and angiotensin II (Ang II) after acute alcohol administration. METHOD: Male Wistar rats were randomly divided into 2 groups: a control group and an ethanol group. The ethanol group intraperitoneally received 3.8 g/kg ethanol; the controls were given the same amount of saline via the same route. After the righting reflex disappeared, midsternotomy was performed in all animals. Immunohistochemical analysis was performed to evaluate protein expression of Cx43 and Ang II. Sections were analyzed by digital image analysis. RESULT: The expression of Cx43 was significantly reduced after acute ethanol treatment, with the integrated optical density lower when compared with control (P < 0.05). The expression of Ang II was significantly increased after acute ethanol treatment, supported by integrated optical density when compared with control (P < 0.05). CONCLUSIONS: In summary, cardiac protein expression of Cx43 and Ang II were found to be significantly altered after acute ethanol treatment, suggesting that these 2 proteins may be important underlying mechanisms of vulnerability to oxidative injury in the heart during acute ethanol. The present study indicated that acute ethanol toxicity caused different alterations in heart proteins that would be related to oxidative stress.

The human and ecological risks of neonicotinoid insecticides in soils of an agricultural zone within the Pearl River Delta, South China.

Neonicotinoid insecticides (NIIs) are extensively used worldwide and frequently detected in the environment. The human and ecological risks associated with the occurrence of NIIs in agricultural zones are of high importance. The present study highlights the regional occurrence and human exposure risks of NIIs in agricultural soil within the Pearl River Delta (PRD), South China. Six neonicotinoids, i.e., imidacloprid, clothianidin, acetamiprid, imidaclothiz, dinotefuran, and flonicamid, were measured in 351 soil samples from Zengcheng, a typical agricultural zone. The soil samples were categorized into three groups based on cultivated plants: vegetables, rice, and fruits. At least one of these neonicotinoid insecticides was detected in 95% of the soil samples. The levels of ∑6NII (range (median)) were 0.26-390 (23), 0.26-280 (6.1), and 0.26-120 (5.0) ng g-1 dry weight in soil samples from vegetable farms, rice paddies, and fruit farms, respectively. Neonicotinoids were detected more frequently and at statistically higher concentrations in vegetable farms than in both rice paddies and fruit farms. This is likely ascribed to higher application frequencies of NIIs in vegetable farms due to higher planting frequencies. The hazard index values for human exposure to NIIs in the agricultural soils were all below 1, suggesting negligible non-cancer risks. The current residual levels of NIIs in the soils could however pose sub-lethal or acute effects to non-target terrestrial organisms such as earthworms. The present study suggests that more information is needed regarding NIIs contamination in soils from agricultural regions of South China to ensure that human and ecological risk from exposure to these compounds can be fully addressed.

[Pharmacognostical studies on root of Geranium strictipes].

OBJECTIVE: To establish pharmacognostical methods on Root of Geranium strictipes. METHODS: To study by the original plant identification, character identification. The organizational structure of root and powder features of this drug were observed and compared. TLC of the drug was also undertaken. RESULTS: It showed that Geranium strictipes powder microstructure contained many calcium oxalate cluster crystals and starch grains. TLC had good reappearance. CONCLUSION: This study provides reference information for further development and identification of this crude drug.

Alterations in ACE and ABCG2 expression levels in the testes of rats subjected to atropine-induced toxicity.

Atropine-induced damage is associated with enzyme and protein alterations. The aim of the present study was to investigate atropine­induced alterations in testicular expression levels of angiotensin­converting enzyme (ACE) and adenosine 5'-triphosphate binding cassette sub­family G member 2 (ABCG2) following atropine treatment. Male Wistar rats received 15 mg/kg/day atropine for 7 days; control rats received an identical volume of saline, Following treatment, the testes were harvested for immunohistochemistry and in situ hybridization to examine the protein and gene expression levels of ACE and ABCG2 by digital image analysis. ACE gene and protein expression levels were significantly reduced in the testes of atropine­treated rats, compared with control rats (P=0.0001 and P<0.001, respectively). In addition, ABCG2 gene and protein expression levels were significantly increased in the testes of atropine­treated rats, compared with control rats (P=0.0017 and P<0.001, respectively). Thus, the results of the present study demonstrate that testicular protein and gene expression levels of ACE and ABCG2 were altered as a result of atropine­induced toxicity in the rats. These alterations may result in abnormal testicular function, and the proteins and genes identified in the present study may be useful to elucidate the mechanisms underlying atropine­induced toxicity and provide a direction for further studies.

Changes in FABP1 and gastrin receptor expression in the testes of rats that have undergone electrical injury.

Testicular trauma may occur due to accidental electrical injury. The aim of this study was to investigate alterations in the levels of fatty acid-binding protein 1 (FABP1) and gastrin receptor (gastrin R) in the testes following electrical injury. Sprague-Dawley rats were divided into control, fatal electrocution (220 V, 50 Hz, 60 sec) and electrical injury (220 V, 50 Hz, 60 sec) groups (n=8 per group). The animals in the fatal electrocution and electrical injury groups were deeply anesthetized with sodium pentobarbital prior to each treatment, in which the current was delivered via an anode connected to the left foreleg and a cathode to the right hindleg. The rats that survived were subsequently sacrificed by cervical dislocation. Control animals received cervical dislocation alone. Immunohistochemical analysis was performed to evaluate the protein expression of FABP1 and gastrin R in the testes. Sections were evaluated by digital image analysis. The expression levels of FABP1 and gastrin R were significantly increased following electrical injury, supported by an increase in the integrated optical density (IOD) when compared with that in the control group (P<0.05). However, no significant difference was found in FABP1 and gastrin R expression levels between the fatal electrocution and control groups. In summary, the protein expression levels of FABP1 and gastrin R were found to be significantly altered by electrical injury, suggesting that these two proteins may be important in underlying mechanisms of testicular injury during electrical injury. The findings indicate that such alterations would be reflected in abnormal testicular function.

[Effects of N and S application on grain filling characteristics and yield of winter wheat].

A field experiment was conducted on the Guanzhong Plain, China, to evaluate the effects of N and S application on characteristics of winter wheat (cv. Xiaoyan 22) grain filling and yield, using a central composite rotatable design with two factors nitrogen (N) and sulfur (S). The results showed that, with the combined application of N and S, grain filling followed a "S" curve which increased slowly at first, then quickly, and then slowly again. With 108 and 267 kg N x hm(-2), the grain fill duration, theoretical maximum grain yield, and average filling rate decreased as the S application rate increased. With 97.5 and 202.5 kg S x hm(-2), N application improved grain filling parameter values. With 187.5 kg N x hm(-2) or 150 kg S x hm(-2), the parameters firstly increased, reached a maximum, and then decreased as the S or N input increased. Grain filling rate increased for 25 days following anthesis, and then declined at a rate that varied among treatments. When the N input was > 187.5 kg x hm(-2) or the S input was > 150 kg x hm(-2), the grain filling rate decreased with increasing the S or N input. The results also indicated that combined application of N and S fertilizers at appropriate rates could increase the head density and grain yield. Using the regression equations, highest grain yields estimates (> or = 3753 kg x hm(-2)) were achieved with the combination of a high N rate (178.31-256.36 kg x hm(-2)) and a moderate S rate (131.95-167.65 kg x hm(-2)).

α-Enolase and caveolin alterations in the heart of rats having undergone gentamicin-induced toxicity.

Gentamicin (GM)-induced heart damage is associated with alterations in expression levels of various enzymes and proteins. The aim of the present study was to investigate GM-induced alterations in cardiac α-enolase and caveolin after GM administration. Male Wistar rats were randomly divided into two groups: a control group and a GM group. The GM group intraperitoneally received GM at a single dose of 7 mg/kg for 8 days, while the controls were given the same amount of saline via the same route. On Day 9, the rats were anesthetized and a thoracotomy was performed in all animals. Immunohistochemical analysis was performed to evaluate protein expression of α-enolase and caveolin. Sections were analyzed by digital image analysis. Our results revealed that cardiac protein expression of α-enolase and caveolin was altered after GM-induced toxicity in the rat. The expression of α-enolase and caveolin was significantly increased after GM-induced toxicity, as determined by integrated optical density analysis, when compared with the control (P<0.05). The current findings indicate that such changes in protein expression may be reflected in abnormal cardiac function, and the proteins identified in this study may be useful for elucidating the mechanisms underlying GM-induced toxicity and may also provide various clues for further investigations.

E-cadherin and caveolin-1 alterations in the heart of rats having undergone chlorpromazine-induced toxicity.

Heart damage induced by chlorpromazine (CPZ) toxicity is associated with changes in the expression of various enzymes and proteins. This study aimed to investigate CPZ­induced alterations in cardiac E-cadherin and caveolin-1 (cav-1) after CPZ administration. Male Wistar rats were randomly divided into two groups: a control group and a CPZ group. The CPZ group was administered CPZ intraperitoneally at a single dose of 10 mg/kg for 21 days; the controls were given the same amount of saline via the same route. On Day 22, the rats were anesthetized, and a thoracotomy was performed in all animals. Immunohistochemical analysis was performed to evaluate protein expression of E-cadherin and cav-1. Sections were analyzed by digital image analysis. Results of the present study revealed that cardiac protein expression of E-cadherin and cav-1 was altered after CPZ-induced toxicity in the rat. The expression of E-cadherin was significantly reduced, while expression of cav-1 was significantly increased after CPZ treatment, as supported by integrated optical density analysis, compared with the control (P<0.05). The current findings indicate that such changes in the expression of E-cadherin and cav-1 may be reflected in abnormal cardiac function, and these proteins may be useful in revealing the mechanisms underlying CPZ-induced toxicity and may also provide additional insight for further research.

E-cadherin and 5-HT alterations in the heart of rats having undergone atropine-induced toxicity.

Atropine-induced heart damage is associated with changes in the expression of various enzymes and proteins. The purpose of this study was to investigate atropine-induced alterations in cardiac E-cadherin and 5-hydroxytryptamine (5-HT) after atropine administration. Male Wistar rats were randomly divided into two groups: a control group and an atropine group. The atropine group intraperitoneally received atropine at a single dose of 15 mg/kg for 7 days; the controls received the same amount of saline via the same route. On Day 8, the rats were anesthetized, and a thoracotomy was performed in all animals. Immunohistochemical analysis was performed to evaluate protein expression of E-cadherin and 5-HT. Sections were analyzed by digital image analysis. Cardiac protein expression of E-cadherin and 5-HT was altered after atropine­induced toxicity in the rat. The expression levels of E-cadherin and 5-HT were significantly decreased after atropine treatment, supported by IOD analysis, when compared with the control (P<0.05). The current findings indicate that such changes would be reflected in abnormal cardiac function, and these proteins may be useful for revealing the mechanisms underlying atropine-induced toxicity and may also provide various clues for further research.

Endothelin-1 and type III collagen alterations in the heart of rats following cisplatin-induced toxicity.

Cisplatin-induced heart damage is associated with enzymes and protein alterations. The purpose of this study was to investigate cisplatin-induced alterations in cardiac endothelin (ET)-1 and type III collagen following administration of cisplatin. Male Wistar rats were randomly divided into two groups: a control and a cisplatin group. The cisplatin group received cisplatin intraperitoneally (i.p.) at a single dose of 7 mg/kg on day 6, while the controls were given the same amount of saline via the same route. On day 11, the rats were anesthetized and a thoracotomy was performed on all animals. Immunohistochemical analysis was performed to evaluate the protein expression of ET-1 and type III collagen. Sections were analyzed by digital image analysis. Results showed that the cardiac protein expression of ET-1 and type III collagen was altered following cisplatin-induced toxicity in rats. The expression of ET-1 and type III collagen was significantly increased after cisplatin treatment, supported by integrated optical density, when compared to the control group (P<0.05). The present findings indicate that such alterations may be reflected in abnormal cardiac function. Additionally, the proteins identified in this study may benefit investigations into the mechanisms underlying cisplatin-induced toxicity, thereby providing the necessary evidence for further research.

Elevated enolase and caveolin-1 in the heart of rats following dexamethasone-induced toxicity.

Dexamethasone (DEX)-induced heart damage is associated with enzyme and protein alterations. The purpose of this study was to investigate DEX-induced alterations in cardiac enolase and caveolin-1 (cav-1) following DEX administration. Male Wistar rats were randomly divided into two groups: a control and a DEX. The DEX group intraperitoneally received DEX at the single dose of 10 mg/kg for 7 consecutive days, and the control was given the same amount of saline via the same route. On day 8, the rats were anesthetized, and a thoracotomy was performed in all animals. Immunohistochemical analysis was performed to evaluate protein expression of enolase and cav-1. Sections were analyzed by digital image analysis. Our results demonstrated that cardiac protein expression of enolase and cav-1 was altered following DEX-induced toxicity in the rat. The expression of enolase and cav-1 was significantly increased after DEX treatment, supported by integrated optical density compared with the control (P<0.05). In conclusion, following DEX-induced toxicity, protein expression of enolase and cav-1 was significantly elevated. The current findings indicate that such alterations would be reflected in abnormal cardiac function, and the proteins identified in this study may be useful in revealing the mechanisms underlying DEX-induced toxicity and also in providing various clues for further research.