Objective To observe the effects ofTanyu Tongzhi Formula on proliferation and migration of human umbilical vein endothelial cells (HUVECs);To explore its activity of pro-angiogenesis.Methods HUVECs were cultured in vitro. Rats were treated with gavage containing different doses ofTanyu Tongzhi Formula (78.0, 39.0, 19.5 g/kg) to prepare serum with different doses of medicine. The activity of HUVECs was measured by MTT method. The effects on migration of HUVECs were inspected by using scratch wound assay.Results Compared with the normal control group, serum containing high-dose and medium-doseTanyu Tongzhi Formula could significantly promote the proliferation of the HUVECs (P<0.01). The rates were 49.78% and 33.92% respectively. And the cell migration rate was higher in the two groups than in the normal control group (P<0.01). The rates were 11.36% and 11.62% respectively.ConclusionTanyu Tongzhi Formula can promote proliferation and migration in HUVEC, which may be active to pro-angiogenesis.

Objective To explore the MRI appearances of heterotopic gray matter. Methods The appearances of MRI of heterotopic gray matter(n=12) confirmed were analyzed retrospectively. Results In the 12 cases,3 lesions were nodular,9 lesions were lamellar. MRI can clearly show the lesions. On T1-WIs and T2-WIs,these lesions appeared isointense to the normal gray matter. 2 lesions associated with arachnoid cyst,2 lesions with schizencephaly and 1 lesion with dysgenesis of the corpus callosum and lipoma of midline. During contrast on MRI(n=5) ,all lesions show unenhancement. Conclusion The MRI have characteristics in diagnosis of heterotopic gray matter and is the best method.

This study was aimed to observe the effect ofRong-Shuan (RS) capsule on rodent tolerance against cerebral ischemia, hypoxia and cerebral reserve capacity, which was related to promote blood circulation and remove blood stasis. Acute cerebral ischemia and anoxia models were established by permanent left carotid artery ligation on C57 BL/6 mice and hypoxia inhalation (O2?N2 = 8?92) for 15 min. Duodenal administration of RS capsule at different doses (100, 200 or 400 mg·kg-1) or saline were given 10 min after ischemia onset. The local brain blood circulation changes and neurobehavioral function were evaluated 24 h after ischemia onset. SD rats were given RS capsule at different doses (75, 150, 300 mg·kg-1) or saline. The effect of RS capsule on improvement of microcirculation disturbance induced by high molecular dextran was observed. The results showed that compared with the sham operation group, the brain blood circulation in the model group was significantly decreased; the cerebral infarction area increased; and the behavioral score after cerebral hypoxia was significantly increased (P < 0.05, orP < 0.01). Meanwhile, after the injection of high molecular dextran among rats in the model group, the cerebral leptomeninx microcirculation was obviously slowed down at 3 timepoints, which were 10, 20 and 30 min. Compared with the model group, RS capsule (400 mg·kg-1) can significantly increase the local blood circulation in the brain of mice, improve behavioral disturbance, reduce cerebral ischemia area (P< 0.05, orP < 0.01). RS capsule can also improve blood flow velocity and flow pattern in cerebral leptomeninx microcirculation disturbance induced by high molecular dextran at different timepoints (P < 0.05, orP < 0.01). It was concluded that RS capsule can increase the tolerance against cerebral ischemia, hypoxia and cerebral reserve capacity in order to protect the neural tissues to promote neuronal recovery.

Objective:To observe the impact of the medicated serum of Shumai Capsule and its decomposing on vascular smooth muscle cells(VSMC) proliferation induced by angiotensinⅡ(AngⅡ) and level of NO,SOD,MDA in cell culture supernatant.Methods:Tissue explant method was used for cultivating vascular smooth muscle cells,serum pharmacology was used,AngⅡ10-7mol/L as a stimulating factor,medicared serum divided into Shumai Capsule group,Huoxue Huayu decomposing group(group 1) and Bupi Yishen decomposing group(group 2),MTT colorimetric was used to test OD values,biochemical detection kit was used to test NO,SOD,MDA levels.Results:The OD value of Shumai Capsule and group1 had significant difference from AngⅡ group and group2(P

Objective To synthesize novel gold nanoparticles of GAL-PEG-GNPs,study its radiation effect on hepatocellular carcinoma cells HepG2 cells in vitro,and investigate the underlying mechanisms.Methods GAL-PEG-GNPs were synthesized and characterized successfully.HepG2 cells were divided into three groups of control,GNPs and GAL-PEG-GNPs.The cytotoxicities of these compounds were tested by the CCK-8 assay and their IC50 values of HepG2 cells were calculated.Cell uptake of nanoparticles was detected by TEM and ICP-MS.The radiosensitization effect of nanoparticles was tested by the colony formation assay.Cell cycle distribution was detected by FCM.The expressions of CAT,SOD,and total GSH were detected with a microplate reader,and the expressions of apoptosis-related proteins were tested by Western blot.Results The GNPs and GAL-PEG-GNPs had absorption peaks at 520 and 530 nm,respectively,and their diameters were (22.6-±2.12) and (32.0 ± 1.41) nm detected by ICP-MS.The GAL-PEG-GNPs and GNPs had similar cytotoxicity profiles (P ＞ 0.05),while GAL-PEG-GNPs could be more effectively uptaken by HepG2 cells than GNPs.The sensitive enhancement ratio (SER) of GNPs and GAL-PEG-GNPs to HepG2 cells were 1.46 和 1.95,respectively.The percentage of cells at phase of G2/M in HepG2 population treated with GNP was higher than that of untreated cells (t =14.20,P ＜0.05).The protein expressions of Cytochrome C,Bax,Caspase-3,and Caspase-9 were upregulated while Bcl-2 expression was down-regulated in the cells treated with GNPs/radiation or GAL-PEGGNPs/radiation.The expressions of CAT,SOD and total GSH in the GNP treated groups were significantly decreased compared with the control group(t =12.34,29.39,12.85,P ＜ 0.05).Conclusions GALPEG-GNPs has obvious radiosensitization effect on HepG2 cells,which is related to the induction of cell apoptosis and the generation of free radicals.

OBJECTIVETo observe the protective effect of serum containing Tanyu Tongzhi formula (TYTZF) on ox-LDL-induced human umbilical vein endothelial cells (HUVECs) injury and detect its mechanism on anti-artherosclerosis.

METHODHUVECs were pretreated with serum containing TYTZF and simvastatin respectively for 2 hours and then added with 100 mg x L(-1) of ox-LDL and laid aside for 24 hours. The activity of HUVECs were measured by the methyl thiazolyl tetrazolium (MTF) method and the NO content in cell culture supernatants were examined by the Griess method. The mRNA levels of Cav-1 and eNOS were measured by the Real-time PCR method. The protein expression of Cav-1 and eNOS were detected by Western blotting.

RESULTThe activity of HUVECs was significantly decreased after ox-LDL treatment (P < 0.01) and this decrease was significantly inhibited by serum containing TYTZF and simvastatin of different doses (P < 0.05). They could enhance the NO content in cell culture supernatants, down-regulate the expression of Cav-1 and up-regulate the expression of eNOS at mRNA and protein levels, which was especially notable after treatment with serum containing TYTZF and simvastatin in large doses.

CONCLUSIONTYTZF has the protective effect on HUVECs by increasing the production of NO and up-regulating Cav-1 expression and down-regulating eNOS expression.

Animals ; Caveolin 1 ; metabolism ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Lipoproteins, LDL ; pharmacology ; Nitric Oxide Synthase Type III ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serum ; chemistry

OBJECTIVETo observe the effect of fluoroacetamide on cardiomyocytes of rat and the antidotal effect of acetamide.

METHODS4 groups of SD rats were treated with various dosages of fluoroacetamid(p.o.) and 2 groups of them were treated with acetamide(i.p.). The changes of cardiomyocytes and serum AST, LDH, CK, CK-MB and HBDH were measured at different intervals after poisoning.

RESULTSIn the group treated with fluoroacetamid 8 mg/kg. bw, serum AST[(589.58 +/- 821.72) U/L], CK[(916.78 +/- 343.55) U/L], HBDH[(504.47 +/- 148.88) U/L] raised obviously compared with control[(187.70 +/- 46.87), (755.65 +/- 498.90), (347.25 +/- 228.40) U/L respectively] (P < 0.01), and the pathological findings such as degeneration, liquefactive necrosis and filtration of inflammatory cells in cardiac muscles were observed 24 hours later, while all the male dead within 3 days. In the group treated with fluoroacetamid 4 mg/kg. bw, serum LDH and HBDH rose significantly compared with control(P < 0.01) 5 day later. On the day of 10, myocardial enzymes restored in all experiment groups with some interstitial fibroblastic proliferation. The pathological changes were reduced in the group treated with acetamide synchronously (100 mg/kg. bw).

CONCLUSIONAcute intoxication of fluoroacetamide could damage cardiomyocytes while acetamide could reduce the injury of them, but the injury was reversible. The levels of serum myocardial enzymes could be a usable index for early diagnosis.

Acetamides ; pharmacology ; Alanine Transaminase ; blood ; Animals ; Antidotes ; pharmacology ; Creatine Kinase, MB Form ; blood ; Fluoroacetates ; toxicity ; L-Lactate Dehydrogenase ; blood ; Myocytes, Cardiac ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVEA sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method has been developed and validated for the determination of brucine and strychnine in rat plasma.

METHODSamples were extracted by ethyl acetate-n-butanol (7: 3). Chromatographic separation was operated on ZORBAX XDB-C18 column with gradient elution of acetonitrile-methanol-water (0.05% acetic acid and 10 nmol x L(-1) ammonium formate contained), followed by LC-MS/MS in positive electrospray ionization. Quantification was carried out on multiple reaction monitoring (MRM) of the transition m/z 395.2/324.2, m/z 335.2/184.2 and m/z 199.1/171.1 for brucine, strychnine and tacrine (internal standard), respectively.

RESULTThe method was linear in the range of 0.195-100 and 0.07840 microg x L(-1) for brucine and strychnine, with coefficient correlation 0.994 and 0.996 respectively. The recoveries of extraction were 78.9% - 102.4% for brucine and 95.2% - 106.1% for strychnine. Precision, accuracy, stability and matrix effect of the analytes met the requirement. The method was applied to a pharmacokinetic study of brucine and strychnine after cutaneous administration of Semen Strychni niosome gel. The C(max) were (26.20 +/- 5.81) and (12.50 +/- 3.00) microg x L(-1) while the AUC(0-infinity), were (193.75 +/- 39.43) and (98.25 +/- 28.54) microg x h x L(-1) of the two components.

CONCLUSIONWe conclude that the niosomes may reduce the systemic exposures and prolong the local release of brucine and strychnine.

Administration, Cutaneous ; Analgesics ; analysis ; pharmacokinetics ; Animals ; Chromatography, Liquid ; Convulsants ; analysis ; pharmacokinetics ; Female ; Gels ; chemistry ; Liposomes ; chemistry ; Male ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Seeds ; chemistry ; Semen ; chemistry ; Specific Pathogen-Free Organisms ; Strychnine ; analogs & derivatives ; analysis ; pharmacokinetics ; Strychnos nux-vomica ; chemistry ; Tandem Mass Spectrometry

Objective: To investigate the effects and mechanisms of Xuebijing injection (XBJ) on Chimeric antigen receptor-T (CAR-T) cell function and its therapeutic potential against CAR-T therapy-associated cytokine storms (CRS). Methods: Anti-CD19 CAR-T cells were established based on FMC63 antibodies. Different doses of XBJ (1 and 10 mg/mL) were added to the culture system. Untreated anti-CD19 CAR-T cells served as negative controls. After 48-h co-culture, the effects of XBJ on CAR-T cell function were assessed. Carboxyfluorescein diacetate succinimidyl ester staining was used to assess the effect of XBJ on CAR-T cell proliferation. Flow cytometry, luciferase reporter gene assays, and real time cellular analysis were employed to evaluate the effects of XBJ on CAR-T cell cytotoxicity in vitro. RNA-sequencing was performed to analyze the effects of XBJ on CAR-T cell gene expression. Network pharmacology predicted potential XBJ therapeutic targets for CRS, which were verified in a THP-1 macrophage inflammation model. Results: XBJ enhanced both the proliferation and tumor killing capacities of CAR-T cells. Transcriptome analysis showed that XBJ treatment affects multiple genes and pathways in CAR-T cells, with differential gene enrichment in multiple cell proliferation and growth factor pathways. Potential targets for CRS control by XBJ were predicted using network pharmacology, and the inhibitory effect of XBJ on the expression of relevant genes was verified using a macrophage model. Conclusion The results of this study indicate that XBJ can enhance the killing effect of CAR-T cells on tumor cells and that the mechanism is related to the regulation of T cell proliferation and activation. Moreover, XBJ inhibited excessive inflammation associated with CAR-T therapy. However, the current findings remain to be further validated through in vivo experiments.

As an essential trace element in living organisms, copper is actively involved in normal physiological processes in various systems and is maintained at low level to achieve copper homeostasis. Copper homeostasis, once being disrupted, would induce cell death, and this new form of cell death is known as copper death. In recent years, copper death has been increasingly recognized as an important factor mediating the onset and progression of central nervous system (CNS) diseases. Therefore, we review the pathogenic mechanism of copper death in CNS diseases, as well as its therapeutic strategies so as to deepen the understanding of researchers.