Modified Erchen decoction ameliorates cognitive dysfunction in vascular dementia rats via inhibiting JAK2/STAT3 and JNK/BAX signaling pathways.

BACKGROUND: Vascular dementia (VaD) is one of the most common clinical syndromes of progressive neurocognitive dysfunction with uncertain mechanisms. Modified Erchen decoction (MECD), developed from "Erchen decoction (ECD)" recorded in "Taiping Huimin Heji Jufang", showed a good effect in the treatment of VaD. However, its therapeutic mechanism is still unclear. PURPOSE: This study aimed to elucidate the multi-target mechanisms of MECD against VaD in vivo and in vitro. METHODS: VaD model was established by two-vessel obstruction (2-VO) in Sprague-Dawley rats. Six groups, including the control, 2-VO operation, MECD treatment (2.5, 5.0 and 10.0 g kg-1 d-1), donepezil hydrochloride (positive control, 0.45 g kg-1 d-1) were designed in the whole experiment. After oral administration for 4 weeks, the effects of MECD were verified by behavioral experiments, histological observation, and biochemical index analysis. The chemical profiling of MECD was performed by UHPLC-Orbitrap Fusion-HRMS, and a "compound-target-pathway" multivariate network was constructed to validate and elucidate its pharmacological mechanisms. RESULTS: Compared with 2-VO group, MECD treatment significantly alleviated anxiety and improved spatial memory in VaD rats according to the open field test (OFT) and Y-maze test. A significant increase in neuron number was observed from hematoxylin and eosin (H&E) stained images in cornu ammonis 1 (CA1) of the hippocampal region after MECD treatment. On the one hand, MECD reduced the plasma levels of triglyceride (TG), low-density lipoprotein (LDL), malondialdehyde (MDA), and amyloid-beta 42 (Aß42), and inhibited mRNA expression of interleukin-1 beta (Il-1ß) and Il-6 in the hippocampus. On the other hand, superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) were significantly increased after treatment with MECD. Moreover, MECD reduced the mRNA expression and protein expression of janus kinase 2 (JAK2), signal transducer and activator of transcription 3 (STAT3), c-Jun N-terminal kinase (JNK), and BCL2-associated X (BAX) in the brain of 2-VO rats. Furthermore, 71 compounds were identified from the extract of MECD. Among them, liquiritin and isochlorogenic acid C gave inhibiting effects on the mRNA expression of Jnk. In addition, liquiritin and hesperetin were conformed with the inhibition of Jak2 transcription level in vitro experiments. CONCLUSION: MECD has demonstrated a significant amelioration effect on cognitive dysfunction in VaD rats via JAK2/STAT3 and JNK/BAX signaling pathways, which represents an innovative insight into the "activate blood and eliminate phlegm" theory.

[Effects of Biochar and Straw Returning on Soil Fungal Community Structure Diversity in Cotton Field with Long-term Brackish Water Irrigation].

Brackish water irrigation increases soil salinity and changes the soil environment, which affects the structure and diversity of soil fungi. In this study, the effects of biochar and straw (3.7 t·hm-2 and 6 t·hm-2, respectively) on soil physical and chemical properties and fungal community structure diversity were investigated on the basis of long-term brackish water irrigation. The results showed that compared to the absence of biochar and straw application (control), biochar application significantly increased pH and the contents of total carbon, available potassium, and available phosphorus in soil but significantly decreased the soil conductivity by 20.71%. Straw treatment significantly increased the content of available potassium and phosphorus but significantly decreased the soil bulk density and conductivity by 4.17% and 64.50%, respectively. The biochar and straw treatment showed an increasing trend in the Chao1 index and ACE index of the fungal community but a decreasing trend in the Shannon index and Simpson index. The dominant fungal phyla in the soil were Ascomycota, Mortierellomycota, Basidiomycota, Chytridiomycota, and Glomeromycota. The dominant fungal genera were Chaetomium, Gibberella, Fusarium, Idriella, and Mortierella. Biochar and straw were applied to increase the relative abundance of Ascomycota, Mortierellomycota, Basidiomycota, Glomeromycota, and Chaetomium. However, the relative abundance of Chytridomycota, Gibberella, and Idriella decreased. LEfSe analysis showed that biochar application and straw returning decreased the number of potential biomarkers in fungal communities. RDA results showed that soil fungal community structure was significantly correlated with EC1:5 and TN. Brackish irrigation had adverse effects on soil, in which EC1:5and TN were the main factors driving the change in soil fungal community structure. The soil fungal community adapted to a salt-stress environment through the improvement of soil by biochar and straw.

Effect of Jinzhen granule on two coronaviruses: The novel SARS-CoV-2 and the HCoV-229E and the evidences for their mechanisms of action.

BACKGROUND: Although severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and human coronavirus 229E (HCoV-229E) pose a huge threat to human public health, no specific treatment is available. Jinzhen granule (JZ) is a traditional eight ingredients-Chinese medicine with prominent efficacy for treating viral-induced diseases. However, little is known about the antiviral effect and mechanism of JZ against SARS-CoV-2 and HCoV-229E. PURPOSE: This study aimed to reveal the antiviral effects of JZ against SARS-CoV-2 and HCoV-229E, and to further explore the underlying mechanisms regulating the host immune response. METHODS: The chromatographic separation of JZ was performed using a Shimadzu analytical high-performance liquid chromatograph with UV detection and Alltech ELSD 2000ES. We conducted cytopathic effect (CPE) and plaque reduction assays to evaluate the antiviral effect of JZ. A lethal human angiotensin converting enzyme 2 (hACE2) transgenic mouse model of SARS-CoV-2 was established to determine the protective effect of JZ on mortality and lung virus titers. Real-time quantitative PCR assays were used to analyze the expression of proinflammatory cytokines in vitro and in vivo. Western blotting was further performed to determine the activities on regulating the nuclear factor kappa B (NF-κB)/MAPK pathway. Finally, mitochondrial membrane potential assays, flow cytometry analysis and western blotting were used to assess the anti-apoptotic potency toward HCoV-229E infection. RESULTS: The results showed that 13 chemical components were identified and five peaks were determined and quantitated (gallic acid 1.97 mg/g, baicalin 20.69 mg/g, glycyrrhizic acid 4.92 mg/g, hyodeoxycholic acid 4.86 mg/g, cholic acid 4.07 mg/g). We found that JZ exerted inhibitory potency against SARS-CoV-2 and HCoV-229E in vitro by using CPE and plaque reduction assays, and it was further found that JZ protected mice infected by SARS-CoV-2 from death and inhibited lung virus titers. JZ also significantly decreased the induction of inflammatory cytokines (IL-1α, IL-6, CCL-5 and MIP-1ß), similar to the observed in vitro effect. Moreover, JZ suppressed the release of inflammatory cytokines in vitro and it decreased the protein expression of p-p38 MAPK, p-JNK, p-NF-κB p65 and p-IκBα induced by HCoV-229E and increased the expression of IκBα. Notably, JZ significantly protected HCoV-229E-infected Huh-7 cells from mitochondrial damage and decreased apoptotic cells. The activation of the mitochondria-mediated apoptotic pathway was inhibited by JZ, as shown by the reduced expression of cleaved caspase-9, caspase-3 and p-PARP. CONCLUSIONS: In conclusion, JZ (gallic acid 1.97 mg/g, baicalin 20.69 mg/g, glycyrrhizic acid 4.92 mg/g, hyodeoxycholic acid 4.86 mg/g, cholic acid 4.07 mg/g) exhibited antiviral activities against SARS-CoV-2 and HCoV-229E by regulating the NF-κB/MAPK pathway and the mitochondria-mediated apoptotic pathway. These findings demonstrated the efficacy of JZ against CoVs and suggested JZ treatment as a novel clinical therapeutic strategy for COVID-19.

[Effects of electroacupuncture on insulin resistance in patients with knee arthroplasty].

OBJECTIVE: To explore the effect of electroacupuncture on the improvement of insulin resistance after knee joint replacement based on the combined spinal and epidural anesthesia and postoperative epidural analgesia. METHODS: Eighty patients with insulin resistance but normal blood glucose were randomly divided into a control group and an observation group, 40 cases in each group.Both groups of patients underwent combined spinal and epidural anesthesia and postoperative epidural analgesia for knee arthroplasty. On the basis of the treatment,electroacupuncture (EA) was applied during the operation and within 1 month after the operation in the observation group, and EA was used at Hegu (LI 4), Qihai (CV 6), Zhongwan (CV 12), Pishu (BL 20), Shenshu (BL 23), etc. once every other day. The control group was not treated with electroacupuncture. Fasting blood glucose and fasting insulin were recorded 30 min before surgery (T0), immediately (T1), 1 d (T2), 3 d (T3), 7 d (T4) and 1 month (T5) after surgery, and the insulin resistance (IR) index was calculated. RESULTS: Compared with those at T0 time point, the IR index of the control group at T1, T3, T4 and T5 time points and the observation group at T1, T4, T5 time points were lower (all P<0.05). Compared with the control group at the same time point, the IR index of the T1 and T5 time points in the observation group was significantly lower (both P<0.05), and it was lower than the insulin resistance standard. CONCLUSION: Combined spinal and epidural anesthesia and postoperative epidural analgesia can improve short-term insulin resistance. Combined with EA, the improvement of insulin resistance is more obviously and longer.

Elucidation of the Intestinal Absorption Mechanism of Loganin in the Human Intestinal Caco-2 Cell Model.

Loganin, iridoid glycosides, is the main bioactive ingredients in the plant Strychnos nux-vomica L. and demonstrates various pharmacological effects, though poor oral bioavailability in rats. In this study, the intestinal absorption mechanism of loganin was investigated using the human intestinal Caco-2 cell monolayer model in both the apical-to-basolateral (A-B) and the basolateral-to-apical (B-A) direction; additionally, transport characteristics were systematically investigated at different concentrations, pHs, temperatures, and potential transporters. The absorption permeability (PappAB) of loganin, which ranged from 12.17 to 14.78 × 10-6cm/s, was high at four tested concentrations (5, 20, 40, and 80µM), while the major permeation mechanism of loganin was found to be passive diffusion with active efflux mediated by multidrug resistance-associated protein (MRP) and breast cancer resistance protein (BCRP). In addition, it was found that loganin was not the substrate of efflux transporter P-glycoprotein (P-gp) since the selective inhibitor (verapamil) of the efflux transporter exhibited little effects on the transport of loganin in the human intestinal Caco-2 cells. Meanwhile, transport from the apical to the basolateral side increased 2.09-fold after addition of a MRP inhibitor and 2.32-fold after addition of a BCRP inhibitor. In summary, our results clearly demonstrate, for the first time, a good permeability of loganin in the human intestinal Caco-2 cell model and elucidate, in detail, the intestinal absorption mechanism and the effects of transporters on iridoid glycosides compounds.

[Effects of auricular point sticking on plasma motilin in patients after gynecological laparoscopic operation under general anesthesia].

OBJECTIVE: To observe the clinical efficacy of auricular point sticking on prevention and treatment of gastrointestinal complications after gynecological laparoscopic operation of general anesthesia, and to explore whether it is achieved by regulating the secretion of plasma motilin (MTL). METHODS: Sixty patients who received selective gynecological laparoscopy under general anesthesia were randomly assigned into an observation group and a control group, 30 patients in each one. The patients in the observation group were treated with auricular point sticking at each morning and night, 30 min before anesthesia, revival after surgery and 24 h after surgery. The adhesive fabric with vaccaria seeds was pressed at shenmen (TF4), wei (CO4), benmen (CO3), jiaogan (AH6a) and pizhixia (AT4) for 3 to 5 min until the sensation of sourness, distension and numb appeared. The treatment was given for one week. The patients in the control group were treated only with similar adhesive fabric at auricular points at identical time points; each auricular point was pressed for 3 to 5 min. The anus exhaust time, defecating time and borborygmus were recorded; the level of plasma MTL was tested 30 min before anesthesia, 24 h after o-peration and 48 h after operation; the occurrence of nausea and vomiting from the end of operation to the end of treatment were also recorded. RESULTS: Compared with the control group, the occurrence of nausea after operation was reduced in the observation group (P<0.05), and the anus exhaust time and defecating time were shortened (both P<0.05), and the recovery of borborygmus was improved (P<0.05). The levels of MTL 24 h and 48 h after surgery were higher than those before operation in the two groups (all P<0.05); The levels of MTL 24 h and 48 h after surgery in the observation group were significantly lower than those in the control group (both P<0.05). CONCLUSIONS: The assist of auricular point sticking could reduce the occurrence of nausea-vomiting and accelerate the recovery of gastrointestinal function in gynecological laparoscopic operation under general anesthesia, which is likely to be related with the inhibition on excess secretion of MTL.

Inhibition of hypoxia and serum deprivation-induced apoptosis by salvianolic acid in rat mesenchymal stem cells.

OBJECTIVE: To investigate the influence and mechanism of salvianolic acid B (SalB) on apoptosis inhibition in rat bone marrow-derived mesenchymal stem cells (BMSCs) induced by hypoxia and serum deprivation (hypoxia/SD). METHODS: SalB concentration of 0.1, 1, 10 or 100 mg/L (drug groups) were investigated for their ability to inhibit apoptosis in rat BMSCs. BMSCs in both the apoptosis model and drug groups were cultured under hypoxic conditions for 6 h, after which cell apoptosis and change in mitochondrial membrane potential (MMP) were detected using flow cytometry. Activation of caspase-3 was detected using western blot analysis. RESULTS: Hypoxia/SD induced apoptosis in rat BMSCs. The early apoptosis rate was lower in the drug groups compared to the apoptosis model group (P < 0.05). SalB was found to inhibit the reduction in MMP and decrease the activation of caspase-3. CONCLUSION: 0.1, 1 and 10 mg/L of SalB inhibits activation of caspase-3 and early apoptosis of rat BMSCs induced by hypoxia/SD and could therefore enhance the survival rate of grafted stem cells.

Current progress of research on pharmacologic actions of salvianolic acid B.

As one of the main water-soluble composites of Radix Salviae, salvianolic acid B is a phenolic acid ingredient of the Chinese drug, which is rich content in the herb and has strong pharmaceutical activity. It is used to treat cardiocerebral vascular diseases, antagonize hepatic/renal fibrosis, prevent cancer, and promote stem cell proliferation and differentiation. In the researches of its acting mechanisms, rather deepened studies have been carried out for its application on cardiocerebral vascular diseases, but that for others are rather fewer.