Evaluation of two deep learning-based approaches for detecting weeds growing in cabbage.

BACKGROUND: Machine vision-based precision weed management is a promising solution to substantially reduce herbicide input and weed control cost. The objective of this research was to compare two different deep learning-based approaches for detecting weeds in cabbage: (1) detecting weeds directly, and (2) detecting crops by generating the bounding boxes covering the crops and any green pixels outside the bounding boxes were deemed as weeds. RESULTS: The precision, recall, F1-score, mAP0.5, mAP0.5:0.95 of You Only Look Once (YOLO) v5 for detecting cabbage were 0.986, 0.979, 0.982, 0.995, and 0.851, respectively, while these metrics were 0.973, 0.985, 0.979, 0.993, and 0.906 for YOLOv8, respectively. However, none of these metrics exceeded 0.891 when detecting weeds. The reduced performances for directly detecting weeds could be attributed to the diverse weed species at varying densities and growth stages with different plant morphologies. A segmentation procedure demonstrated its effectiveness for extracting weeds outside the bounding boxes covering the crops, and thereby realizing effective indirect weed detection. CONCLUSION: The indirect weed detection approach demands less manpower as the need for constructing a large training dataset containing a variety of weed species is unnecessary. However, in a certain case, weeds are likely to remain undetected due to their growth in close proximity with crops and being situated within the predicted bounding boxes that encompass the crops. The models generated in this research can be used in conjunction with the machine vision subsystem of a smart sprayer or mechanical weeder. © 2024 Society of Chemical Industry.

Dopaminergic System in Promoting Recovery from General Anesthesia.

Dopamine is an important neurotransmitter that plays a biological role by binding to dopamine receptors. The dopaminergic system regulates neural activities, such as reward and punishment, memory, motor control, emotion, and sleep-wake. Numerous studies have confirmed that the dopaminergic system has the function of maintaining wakefulness in the body. In recent years, there has been increasing evidence that the sleep-wake cycle in the brain has similar neurobrain network mechanisms to those associated with the loss and recovery of consciousness induced by general anesthesia. With the continuous development and innovation of neurobiological techniques, the dopaminergic system has now been proved to be involved in the emergence from general anesthesia through the modulation of neuronal activity. This article is an overview of the dopaminergic system and the research progress into its role in wakefulness and general anesthesia recovery. It provides a theoretical basis for interpreting the mechanisms regulating consciousness during general anesthesia.

Dexmedetomidine mitigates neuroinflammation and improves early postoperative neurocognitive dysfunction in rats by enhancing autophagy.

Postoperative neurocognitive dysfunction (PND) is a common postoperative complication. Autophagy is correlated with the pathogenesis of PND. This study investigated the potential role of autophagy in the neuroprotection of dexmedetomidine (Dex) pretreatment in PND. The PND rat model was established by abdominal surgery. The cognitive function of rats was evaluated by Y-maze 3 days after surgery. Nissl staining assessed postoperative hippocampal damage. Immunofluorescence detected the expression of microglial activation (Iba-1) and autophagy-related protein (LC3B) in hippocampal tissues. Western blot detected the autophagy-related protein expression (Beclin 1, LC3B, and p62), proinflammatory cytokines, and the protein activation of the autophagy-related LKB1/AMPK/ULK-1 signaling pathway. RT-PCR quantified the expression of IL-1ß, TNF-α, and IL6. In this study, we found that Dex pretreatment improved spatial memory function impairment and reduced abdominal surgery-induced hippocampal tissue damage. Dex pretreatment significantly increased the expression of Beclin 1 and LC3 II/I and decreased the expression of p62 in the hippocampus after surgery. Furthermore, Dex effectively inhibited microglial activation and proinflammatory cytokines by enhancing autophagy in the hippocampus. Pretreatment with 3-MA, an autophagy inhibitor, significantly weakened the inhibitory effect of Dex on postoperative neuroinflammation. We further demonstrated that Dex suppressed surgery-induced neuroinflammation by activating the LKB1/AMPK/ULK-1 signaling pathway. In conclusion, our study indicated that Dex inhibited hippocampal neuroinflammation and ameliorated PND by enhancing autophagy after surgery in rats, which was related to the LKB1/AMPK/ULK-1 signaling pathway. These findings provide a potential therapeutic prospect for PND.NEW & NOTEWORTHY Dex inhibits hippocampal neuroinflammation and attenuates early cognitive impairment by enhancing autophagy following surgery in rats. Dex may protect postoperative cognitive function by activating the LKB1/AMPK/ULK-1 signaling pathway.

Isolation and characterization of antioxidant peptides from oyster (Crassostrea rivularis) protein enzymatic hydrolysates.

Peptides from oysters have several bioactive functions. In this study, we identified antioxidant peptides from oysters (Crassostrea rivularis) and investigated their structure-function relationship. We used an 8 kDa molecular-weight (MW) cut-off membrane and semiprep reversed-phase liquid chromatography to collect five peptides (F1-F5) and identified the highest-abundance ion-peak sequences AWVDY (F1), MSFRFY(F2), EPLRY(F3), RKPPWPP(F4), and YAKRCFR(F5) having MWs of 652, 850, 676, 877, and 943 Da, respectively, using ultra-performance liquid chromatography-quadrupole/time-of-flight tandem mass spectrometry. These peptides exhibited high antioxidant activities, similar to butylated hydroxytoluene, reduced glutathione, and ascorbic acid. F5 demonstrated the highest scavenging activity for DPPH radicals (IC50 = 21.75 µg/ml), hydroxyl radicals (IC50 = 18.75 µg/ml), and superoxide radicals (IC50 = 11.00 µg/ml), while F3 demonstrated the highest reducing power. Furthermore, F5 significantly protected Caco-2 cells from H2O2-induced oxidative damage. These results suggest that the antioxidant peptide F5 is a promising food additive that protects against oxidative damage.

Analgesic Efficacy of Quadratus Lumborum Block in Patients Undergoing Nephrectomy: A Systematic Review and Meta-Analysis.

OBJECTIVE: To evaluate the analgesic efficacy of quadratus lumborum block (QLB) in adults undergoing nephrectomy. DESIGN: Systematic review and meta-analysis. PATIENTS: Adult patients (≥18 years of age) received nephrectomy under general anesthesia. METHODS: We searched PubMed, Embase, the Cochrane Library, and Web of Science on January 10, 2022, including randomized controlled trials that evaluated the analgesic efficacy of QLB for patients undergoing nephrectomy. RESULTS: A total of 12 randomized controlled trials (N = 821 patients) were included in the study. Compared with the non-block, single-shot QLB reduced postoperative opioid consumption (mean difference [MD], -8.37 mg intravenous morphine equivalent; 95% confidence interval [CI], -12.19 to -4.54 mg) and pain scores at 2 hours, 6 hours, 12 hours, and 24 hours at rest and during movement after nephrectomy. Single-shot QLB also prolonged the time to first analgesic request (MD, 6.44 hours; 95% CI, 2.23 to 10.65 hours), shortened the length of hospital stay (MD, -0.32 day; 95% CI, -0.55 to -0.09 day), and decreased the incidence of postoperative nausea and vomiting (risk ratio, 0.48; 95% CI, 0.36 to 0.65). Compared with continuous epidural anesthesia, repeated QLB could provide comparable postoperative analgesic benefits. CONCLUSIONS: Single-shot QLB provided a statistically significant but clinically small improvement in postoperative analgesia and recovery for patients undergoing nephrectomy. The QLB would be beneficial as part of multimodal analgesia. Future research might need to determine which approach of QLB is superior for postoperative analgesia after nephrectomy.

Electroacupuncture Alleviates Neuroinflammation by Inhibiting the HMGB1 Signaling Pathway in Rats with Sepsis-Associated Encephalopathy.

Sepsis-Associated Encephalopathy (SAE) is common in sepsis patients, with high mortality rates. It is believed that neuroinflammation is an important mechanism involved in SAE. High mobility group box 1 protein (HMGB1), as a late pro-inflammatory factor, is significantly increased during sepsis in different brain regions, including the hippocampus. HMGB1 causes neuroinflammation and cognitive impairment through direct binding to advanced glycation end products (RAGE) and Toll-like receptor 4 (TLR4). Electroacupuncture (EA) at Baihui (GV20) and Zusanli (ST36) is beneficial for neurological diseases and experimental sepsis. Our study used EA to treat SAE induced by lipopolysaccharide (LPS) in male Sprague-Dawley rats. The Y maze test was performed to assess working memory. Immunofluorescence (IF) and Western blotting (WB) were used to determine neuroinflammation and the HMGB1 signaling pathway. Results showed that EA could improve working memory impairment in rats with SAE. EA alleviated neuroinflammation by downregulating the hippocampus's HMGB1/TLR4 and HMGB1/RAGE signaling, reducing the levels of pro-inflammatory factors, and relieving microglial and astrocyte activation. However, EA did not affect the tight junctions' expression of the blood-brain barrier (BBB) in the hippocampus.

Sedative effect of remimazolam combined with alfentanil in colonoscopic polypectomy: a prospective, randomized, controlled clinical trial.

BACKGROUND: Remimazolam is a newer benzodiazepine with properties of rapid onset, short duration of action, and fast recovery. Our study was to evaluate the effects of different doses of remimazolam combined with alfentanil in colonoscopic polypectomy. METHODS: One hundred twenty patients were randomly divided into four groups: alfentanil and propofol (AP) group, alfentanil and remimazolam 0.1 mg/kg (AR1 group), 0.15 mg/kg (AR2 group), or 0.2 mg/kg (AR3 group). Patients in the four groups received alfentanil 10 µg/kg, followed by propofol 2 mg/kg and three dosages of remimazolam. Modified Observer's Assessment of Alertness and Sedation (MOAA/S) scale, heart rate (HR), oxygen saturation (SpO2), respiratory rate (RR), bispectral index (BIS) values and mean arterial pressure (MAP) were collected at intervals of 5 min and analyzed at different time points: before anesthesia (T0), 5 min (T1), 10 min (T2), 15 min after anesthesia (T3) and at the end of surgery (T4). The average MAP was calculated utilizing the average of all MAP values. The primary outcome was the success rate of sedation. Secondary outcomes included time to full alert and adverse events. RESULTS: The success rate of sedation was 100% among the four groups. The incidence of hypotension was significantly decreased (all P < 0.05) and the average MAP was higher in AR1-AR3 groups than AP group (all P < 0.001). None of the patients developed bradycardia or hypertension during surgery in all study groups. BIS values were higher (all P < 0.001) and the time to full alert was statistically shorter in AR1-AR3 groups (all P < 0.05) compared with the AP group. The MOAA/S score in AR1 was higher than AR2 (P < 0.05) and the AR3 group (P < 0.05) at T1 and BIS values in the AR1 group were significantly higher than AR3 group (P < 0.05) at T4. CONCLUSIONS: Remimazolam combined with alfentanil have a non-inferior sedative effect than propofol during the colonoscopic polypectomy. Moreover, this combination of two short-acting drugs might be a safer alternative. TRIAL REGISTRATION: The clinical trial was registered on (16/05/2021, ChiCTR2100046492).

Evaluation of prediction effect of perfusion index for supraclavicular brachial plexus block in children: protocol for a randomized trial.

BACKGROUND: Pulse perfusion index (PI) reflects blood perfusion. It has been reported that PI can be used to evaluate the effect of nerve block, but currently, it is mainly focused on awake adults. In pediatric general anesthesia, it has been reported that PI can evaluate the effect of the sacral block. Still, there is a lack of relevant research on the impact of brachial plexus blocks. Our objective is to assess the prediction effects of PI on the success of supraclavicular brachial plexus block in pediatric patients under sevoflurane or propofol general anesthesia. METHODS/DESIGN: This is a mono-center, parallel, 2-arm randomized superiority trial. One hundred four children aged 1 month to 12 years who undergo upper limb surgery will be enrolled in this study. According to anesthesia induction and maintenance medication, they will be divided into sevoflurane and propofol groups. The PI values of the index and little finger will be recorded on the blocked and non-blocked sides of supraclavicular brachial plexus block (SCB) in all children. The primary outcome is to assess the effects of PI on the success of supraclavicular brachial plexus block in pediatric patients under sevoflurane or propofol general anesthesia. The secondary outcome includes mean arterial blood pressure (MAP), heart rate (HR), and correlation between baseline PI and 10 min after SCB (PI ratio). DISCUSSION: This trial will provide evidence on the changes in PI after SCB in sevoflurane or propofol anesthesia in children. SCB may lead to changes in PI values under sevoflurane or propofol anesthesia. After the children wake up at the end of the surgery, the changes in PI values on the block side and non-block side may be helpful to judge the effect of nerve block when excluding the influence of anesthetics. TRIAL REGISTRATION: ClinicalTrials.gov NCT04216823 . Registered on 15 July 2020.

Blocking the Aryl Hydrocarbon Receptor Alleviates Myocardial Ischemia/Reperfusion Injury in Rats.

OBJECTIVE: Restoring the blood perfusion of ischemic heart tissues is the main treatment for myocardial ischemia. However, the accompanying myocardial ischemia reperfusion injury (IRI) would aggravate myocardial damage. Previous studies have confirmed that aryl hydrocarbon receptor (AhR) is closely correlated to kidney and intestinal IRI. The present study aimed to explore the relationship between AhR and myocardial IRI. METHODS: An oxygen glucose deprivation/reoxygenation (OGD/R) model of H9c2 cells and an ischemia/reperfusion (I/R) model of Sprague-Dawley rat myocardium were established. OGD/R cells and myocardial IRI rats were treated with different concentrations of the AhR antagonist CH-223191 or agonist 6-formylindolo[3,2-b] carbazole (FICZ). Under the conditions of normoxia and hypoxia/reoxygenation, the activity of cardiomyocytes, lactate dehydrogenase (LDH) and cell reactive oxygen species (ROS) were detected. In rats, myocardial pathological damage and markers of myocardial injury were detected. RESULTS: According to the results of the cell viability, LDH and ROS tests in vitro, both CH-223191 and FICZ showed no myocardial protection under OGD/R conditions. However, the histological staining and analysis of myocardial injury marker LDH in vitro revealed that CH-223191 could significantly reduce the myocardial IRI. CONCLUSION: AhR exhibited a different effect on myocardial IRI in vitro and in vivo. In vivo, CH-223191 could significantly alleviate the myocardial IRI, suggesting that inhibition of AhR may play a role in myocardial protection, and AhR may serve as a potential treatment target for myocardial IRI.

Dexmedetomidine attenuates perioperative neurocognitive disorders by suppressing hippocampal neuroinflammation and HMGB1/RAGE/NF-κB signaling pathway.

Surgical trauma can induce an inflammatory response in the central nervous system. Neuroinflammation is a crucial pathological mechanism of perioperative neurocognitive disorders (PND). Dexmedetomidine (Dex) is an alpha (α)-2 adrenoceptor agonist that is widely used in the perioperative period. Previous studies have shown that Dex has neuroprotection in various nerve injury models, but its role in PND remains unclear. Our study aimed to observe the neuroprotective effect of Dex pretreatment on postoperative cognitive change and explore the effects of hippocampal neuroinflammation, microglial polarization and HMGB1/RAGE/NF-κB signaling pathway involved in Dex on PND in rats. Rats were pretreated with Dex alone or in combination with yohimbine (α-2 adrenoceptor antagonist) before surgery. Behavioral tests results showed that Dex ameliorated surgery-induced cognitive impairment in rats. Nissl, immunohistochemistry and TUNEL-NeuN staining results indicated that Dex reduced hippocampus damage and neuronal apoptosis caused by surgery. Dex preconditioning reduced the expression of the proinflammatory cytokines IL-1ß, TNF-α and IL-6 in hippocampus. Immunohistochemical and immunofluorescence results showed that Dex preconditioning inhibited the activation of glial cells induced by surgery. Western blot analysis showed that Dex preconditioning downregulated the expression of M1 phenotype markers (CD86 and iNOS), HMGB1, RAGE and nuclear NF-κB and upregulated the expression of M2 phenotype markers (Arginase 1 and CD206) and cytoplasmic NF-κB. Yohimbine could inhibit the neuroprotective effect of Dex. These results indicated that Dex pretreatment could improve postoperative short-term cognitive impairment, and the neuroprotective mechanism may involve the suppression of hippocampal neuroinflammation, regulation of M1/M2 polarization, and inhibition of HMGB1/RAGE/NF-κB signal transduction.

Electroacupuncture ameliorates neuroinflammation in animal models.

BACKGROUND: Neuroinflammation refers to a wide range of immune responses occurring in the brain or spinal cord. It is closely related to a variety of neurodegenerative diseases, for which it potentially represents a new direction for treatment. Electroacupuncture (EA) is one method of acupuncture treatment, which can be used as an adjuvant therapy for many diseases. This review focuses on molecular mechanisms of EA in the reduction of neuroinflammation, summarizes relevant basic research and outlines future directions for investigation. FINDINGS: A growing body of basic research has shown that EA can ameliorate neuroinflammation centrally (in animal models of ischemic stroke, Alzheimer's disease, traumatic brain injury, spinal cord injury, Parkinson's disease and vascular dementia) and peripherally (e.g. after a surgical insult or injection of lipopolysaccharide) and that its effects involve different molecular mechanisms, including activation of the α7 nicotinic acetylcholine receptor signaling pathway and P2 type purinergic receptors, inhibition of nuclear factor κB, and mitigation of damage secondary to oxidative stress and NOD-like receptor protein 3 inflammasome activation. CONCLUSIONS: EA is capable of regulating multiple cell signal transduction pathways to alleviate neuroinflammation in animal models. Although the findings of animal studies are encouraging, further prospective clinical trials are needed to verify the efficacy of EA for the treatment of neuroinflammation.

Application of UHPLC-Q/TOF-MS-based metabolomics in the evaluation of metabolites and taste quality of Chinese fish sauce (Yu-lu) during fermentation.

Spontaneous fermentation is a critical step in the processing of high-quality fish sauce. In this study, a comparative UHPLC-Q/TOF-MS-based metabolomics approach combining equivalent-quantification and the taste activity value (TAV) was used, for the first time, to evaluate the taste qualities and characterize metabolite profiles in Chinese fish sauce during fermentation. A total of 22,816 metabolite ion features were extracted from fish sauce samples. Forty-six metabolites, including amino acids, small peptides, organic acids, amines, carbohydrates, and nucleic acids, were identified as key chemical components of fish sauce. In addition, absolute quantification and TAV showed that aspartic acid and glutamic acid exert an important influence on the umami taste of fish sauce. Specific metabolites were primarily associated with amino acid metabolism, particularly alterations in arginine and proline metabolism. This study identifies chemical components and provides novel insights into the taste quality of fish sauce due to fermentation.

Effect of Ultrastructure on Changes of Textural Characteristics between Crisp Grass Carp (Ctenopharyngodon Idellus C.Et V) and Grass Carp (Ctenopharyngodon Idellus) Inducing Heating Treatment.

The research studies the ultrastructure effect on texture of crisp grass carp (CGC) and grass carp (GC) fillets inducing heating for 15, 25, and 40 min with boiling water. After heating, the hardness, fracturability, springiness, chewiness, resilience, and cohesiveness of CGC were higher than that of raw CGC, whereas the all textural characteristics of heating GC were lower obviously than that of raw GC. The hardness, fracturability, springiness, chewiness, resilience, and cohesiveness of CGC for heating 15 min were higher by 6.3%, 9.0%, 27.0%, 71.8%, 9.4%, and 23.9%, respectively, than that of raw CGC (RCGC). The hardness increasing of CGC flesh with the extension of heating time related closely to more coagulating connective tissue in interstitial spaces, especially relating to smaller muscle fiber diameter and denser muscle fiber density. The more and larger spaces between fiber and fiber with the extension of heating time results in the decrease of cohesiveness and resilience of CGC flesh. For chewiness, the stronger chewiness of cooked CGC associated with more detachment of myofiber-myocommata and fiber-fiber. Overall, the results show that the changes of texture characteristics of CGC fillet with extension of heating time correlates positively with the ultrastructure.

Bioactive components on immuno-enhancement effects in the traditional Chinese medicine Shenqi Fuzheng Injection based on relevance analysis between chemical HPLC fingerprints and in vivo biological effects.

ETHNOPHARMACOLOGICAL RELEVANCE: Shenqi Fuzheng Injection (SFI) is an injectable traditional Chinese herbal formula comprised of two Chinese herbs, Radix codonopsis and Radix astragali, which were commonly used to improve immune functions against chronic diseases in an integrative and holistic way in China and other East Asian countries for thousands of years. MATERIALS AND METHODS: This present study was designed to explore the bioactive components on immuno-enhancement effects in SFI using the relevance analysis between chemical fingerprints and biological effects in vivo. According to a four-factor, nine-level uniform design, SFI samples were prepared with different proportions of the four portions separated from SFI via high speed counter current chromatography (HSCCC). SFI samples were assessed with high performance liquid chromatography (HPLC) for 23 identified components. For the immunosuppressed murine experiments, biological effects in vivo were evaluated on spleen index (E1), peripheral white blood cell counts (E2), bone marrow cell counts (E3), splenic lymphocyte proliferation (E4), splenic natural killer cell activity (E5), peritoneal macrophage phagocytosis (E6) and the amount of interleukin-2 (E7). Based on the hypothesis that biological effects in vivo varied with differences in components, multivariate relevance analysis, including gray relational analysis (GRA), multi-linear regression analysis (MLRA) and principal component analysis (PCA), were performed to evaluate the contribution of each identified component. RESULTS: The results indicated that the bioactive components of SFI on immuno-enhancement activities were calycosin-7-O-ß-d-glucopyranoside (P9), isomucronulatol-7,2'-di-O-glucoside (P11), biochanin-7-glucoside (P12), 9,10-dimethoxypterocarpan-3-O-xylosylglucoside (P15) and astragaloside IV (P20), which might have positive effects on spleen index (E1), splenic lymphocyte proliferation (E4), splenic natural killer cell activity (E5), peritoneal macrophage phagocytosis (E6) and the amount of interleukin-2 (E7), while 5-hydroxymethyl-furaldehyde (P5) and lobetyolin (P13) might have negative effects on E1, E4, E5, E6 and E7. Finally, the bioactive HPLC fingerprint of SFI based on its bioactive components on immuno-enhancement effects was established for quality control of SFI. CONCLUSIONS: In summary, this study provided a perspective to explore the bioactive components in a traditional Chinese herbal formula with a series of HPLC and animal experiments, which would be helpful to improve quality control and inspire further clinical studies of traditional Chinese medicines.

Network pharmacology analyses of the antithrombotic pharmacological mechanism of Fufang Xueshuantong Capsule with experimental support using disseminated intravascular coagulation rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Fufang Xueshuantong (FXST) Capsule is developed on a traditional Chinese medicine remedy, with a four-herb formula of Panax notoginseng, Radix astragali, Salvia miltiorrhizae and Radix scrophulariaceae. It has been used for treatment of the clinic cardiovascular disease for many years. MATERIALS AND METHODS: Due to its complexity of compositions and polypharmacological effects, it often complicates understanding of the mechanisms of action. In the present work, we have constructed an integrated model of system pharmacology to investigate the polypharmacological mechanisms of FXST formulation for treatment of thrombosis disease. RESULTS: The predicted results showed that 22 ingredients in FXST were closely associated with 41 protein targets related to blood coagulation, fibrinolysis and platelet aggregation. Through analysis of the compound-protein target association, significant cross-targets between each herb indicated the multiple active chemical ingredients might interact with the same target simultaneously and thus explained the synergistic mechanisms of the principle of Traditional Chinese medicines (TCMs) as ''Jun (emperor) - Chen (minister) - Zuo (adjuvant) - Shi (courier)''. To validate the polypharmacological effects predicted by our network pharmacology (NetPharm) analysis, we have carried out experimental investigation the effects of FXST on the disorders of the blood coagulation system in a lipopolysaccharide-induced disseminated intravascular coagulation (DIC) rat model. The results showed that FXST could significantly ameliorate the activation of coagulation system, which is congruent with the cross-target prediction by NetPharm approach. CONCLUSIONS: The combined investigations provide more insight into better understanding of the pharmacological mechanisms of FXST, and may also offer an alternative avenue to further explore the chemical and pharmacological basis of TCMs.

Rapid separation and identification of multiple constituents in traditional Chinese medicine formula Shenqi Fuzheng Injection by ultra-fast liquid chromatography combined with quadrupole-time-of-flight mass spectrometry.

Shenqi Fuzheng Injection (SFI) a well-known traditional Chinese medicine (TCM) formula, has been extensively used as an adjuvant to chemotherapy for cancer treatment in clinic. However, the chemical constituents in SFI, especially water-soluble ingredients, had not been investigated so far. In this study, an ultra-fast liquid chromatography (UFLC) coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS/MS) method was established for rapid separation and structural identification of the constituents in SFI. Separation was performed on a C18 reversed-phase column (2.1 mm × 100 mm, 1.8 µm) by gradient elution mode, using methanol-water containing 0.1% formic acid as mobile phase at the flow-rate of 0.2 mL/min. Accurate mass measurement for molecular ions and characteristic fragment ions could represent reliable identification criteria for these compounds. As a result, eighty-one major constituents including organic acids, amino acids, oligosaccharides, alkaloids, nucleosides, phenylpropanoids, polyacetylenes, flavonoids, isoflavonoids and saponins were identified or tentatively characterized by comparing their retention times and MS spectra with those of authentic standards or literature data. All compounds were further assigned in the individual raw material. In conclusion, the UFLC-Q-TOF-MS/MS is a highly efficient technique to separate and identify constituents in complex matrices of traditional Chinese medicines. These results obtained in this research will provide a basis for quality control and further study in vivo of SFI.

Immuno-enhancement effects of Shenqi Fuzheng Injection on cyclophosphamide-induced immunosuppression in Balb/c mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Radix Codonopsis and Radix Astragali, of which Shenqi Fuzheng Injection (SFI) is composed, are commonly used in traditional Chinese medicine to improve immune function against chronic diseases. AIM OF THE STUDY: The present study was thus designed to systematically elucidate the in vivo immuno-enhancement effects of SFI in immunosuppressed mice induced by cyclophosphamide (Cy) treatment. MATERIALS AND METHODS: Balb/c mice were injected intraperitoneally (i.p.) once daily with low-dose (2.5 g raw materials/kg), intermediate-dose (5 g raw materials/kg), high-dose (10 g raw materials/kg) of SFI for 10 consecutive days, respectively, accompanied by i.p. injection of Cy (80 mg/kg) on Days 4-6. RESULTS: Compared with vehicle group, low-, intermediate- and high-dose SFI treatment accelerated recovery dose-dependently of spleen index, peripheral white blood cell and bone marrow cell counts, enhanced T cell and B cell proliferation responses, as well as splenic nature killer cell activity and peritoneal macrophage phagocytosis, and restored the level of interleukin-2 in the serum. Furthermore, SFI treatment promoted recovery of the amount of peripheral white blood cells on Day 6, rather than recombinant human Granulocyte Colony-Stimulating Factor (rhG-CSF) did. CONCLUSIONS: These results demonstrate, for the first time, that chronic treatment with SFI results in accelerating recovery of immunosuppression in Cy-treated mice, which is competent in taking into consideration for both precautions and remedy. Our findings provide experimental evidences for further researches and clinical application in cancer patients undergoing chemotherapy.

Lipid composition and grafted PEG affect in vivo activity of liposomal mitoxantrone.

Mitoxantrone was encapsulated into pegylated SUVs using ammonium sulfate gradient method. Four formulations (LM-s, LM-p, LM-m and LM-m-L) were prepared, which were made from different PCs and exhibited different PEG grafting density. In vitro release studies revealed that drug release rate increased with decreased T(m) of PCs, and reduced PEG polymer coverage. In circulation, the trend towards increased circulation time as T(m) of PCs and PEG lipid content are elevated is observed. However, it was found that the order of toxicity in balb/c mice was Lm-s

Encapsulation of mitoxantrone into pegylated SUVs enhances its antineoplastic efficacy.

Mitoxantrone (MIT) was encapsulated into 60, 80 and 100nm pegylated hydrogenated soy phosphatidylcholine/cholesterol (HSPC/chol) vesicles using a transmembrane (NH(4))(2)SO(4) gradient. In-vitro release studies revealed that small-sized formulation had fast drug-release rate. Acute toxicity studies performed in c57 mice proved that all pegylated liposomal MIT (plm) formulations could be well-tolerated at a dose of 9mg/kg, significantly compared to severe toxicity induced by free mitoxantrone (f-M). In KM mice, plm60 was at least 2- to 3-fold less toxic than f-M. After intravenous injection, plm60 was slowly eliminated from plasma relative to f-M, resulting in about 6459-fold increase in AUC and its plasma kinetics exhibited dose dependence. In S-180 bearing KM mice, plm60 preferentially accumulated into tumor zone, with a approximately 12-fold increase in AUC and approximately 10-fold increase in C(max) Furthermore, the accumulation of plm60 in almost all normal tissues markedly decreased. The antitumor efficacy of plm60 was also considerably enhanced. In L1210 ascitic tumor model, plm60 was the most efficacious which led to a approximately 70% long-term survival, significantly compared to 16-33% survival rate in plm80, plm100 and f-M groups at the same dose level (4mg/kg). The antitumor efficacy of plm60 was more encouraging in L1210 liver metastasis model. At a dose of 6mg/kg, approximately 90% animals receiving plm60 treatment could survive 60 days; however, in f-M group at the same dose, all the mice died at approximately 14 days post inoculation. Similarly, plm60 could effectively inhibit the growth of RM-1 tumor in BDF1 mice, resulting in marked increase in tumor doubling time at different dose levels relative to f-M. The improved antineoplastic effects could be ascribed to its small vesicle size, which allowed more drug release after the accumulation into tumor zone. Theoretical considerations revealed that the reduction of vesicle size could increase the specific area of MIT/sulfate precipitate inside the vesicle and the release constant K, which is inversely proportional to vesicle volume (K=pA(m)k(2)k(2)(')/([H(+)](i)(2)V(i))).

Copper ion-mediated liposomal encapsulation of mitoxantrone: the role of anions in drug loading, retention and release.

Besides pH gradient, other transmembrane gradients such as metal ion gradient could be also employed to load drugs into liposomes. In pH gradient method, anions have an important role since they could form specific aggregates with drugs, and then affect drug release kinetics from vesicles. To explore the role of anions in metal ion gradient method, copper ion-mediated mitoxantrone (MIT) loading was investigated systematically. When empty liposomes exhibiting a transmembrane copper ion gradient (300 mM) were mixed with MIT in a molar ratio of 0.2:1, after 5 min incubation at 60 degrees C, >95% MIT could be loaded into vesicles and the encapsulation was stable, regardless of the kinds of anions and initial intraliposomal pH values. The encapsulation ratio decreased with increased MIT/lipid molar ratio. But even when the molar ratio increased to 0.4, >90% encapsulation could still be achieved. In the presence of nigericin and ammonium, the drug loading profiles were affected to different degree with respect to both drug loading rate and encapsulation ratio. Relative to CuSO(4)-containing systems, CuCl(2) mediated MIT loading was unstable. Both nigericin and ammonium could alter the absorption spectra of liposomal MITs loaded with CuSO(4) gradient. In vitro release studies were performed in glucose/histidine buffer and in 50% human plasma using a dialysis method. In both of release media, CuCl(2)-containing vesicles displayed rapid release kinetics in comparison with CuSO(4) systems; and during the experiment period, MIT was lost from the vesicles continuously. When the formulations were injected into BDF1 mice at a dose of 4 mg/kg, all the liposomal formulations exhibited enhanced blood circulation time, with half-life values of 6.8-7.2h, significantly compared to the rapid clearance of free-MIT. In L1210 ascitic model, CuCl(2) formulation was more therapeutically active than CuSO(4) formulation. At a dose of 6 mg/kg, the treatment with CuCl(2) formulation resulted in a median survival time of 21 days, considerably larger than that of CuSO(4) groups (15 days). Based on these data, it was concluded that during the drug loading process, a dynamic transmembrane pH gradient is generated and intraliposomal pH might affect the complexation manner in which Cu(2+) binds MIT. Owing to the presence of pH gradient, after the accumulation within vesicles, a part of MIT will be protonated and precipitated by sulfate. Accordingly, the aggregation status of MIT inside CuSO(4) system was more complicated than that in CuCl(2) vesicles. The difference in physical status of MIT aggregates affects not only the drug release rate, but also their therapeutic effects.