A novel strategy for active compound efficacy status identification in multi-tropism Chinese herbal medicine (Scutellaria baicalensis Georgi) based on multi-indexes spectrum-effect gray correlation analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Bioactive substance identification is always the focal point and the main challenge in Chinese herbal medicine (CHM). Most CHM present multiple efficacies and multiple tropisms, which has improved the application accuracy of CHM, and is worthy of further study. In this article, the concept of "multi-tropism efficacy of CHM" has been proposed for the first time. In addition, it is hypothesized that the different components in CHM can be classified based on their efficacy status. AIM OF THE STUDY: The spectrum-effect relationship between the fingerprint and efficacy was established to identify the efficacy status of components. This provided a practical, efficient and accurate way to identify the bioactive substances from a complex CHM system. MATERIALS AND METHODS: The network pharmacology approach was applied to preliminarily analyze the potential antibacterial compounds and mechanisms of HQ. Furthermore, its chemical fingerprint was established and the characteristic peaks were identified by LC-MS/MS. The antibacterial and anti-inflammatory bioactivities of HQ were determined to evaluate its pharmacological effect of heat-clearing and detoxification, and its anticoagulation activity was determined to evaluate its heat-clearing and tocolysis effects. The spectrum-effect relationships were assessed by gray correlation analysis to discriminate the status of active components in HQ with different efficacies. RESULTS: Network pharmacology analysis revealed apigenin, wogonin, baicalein, acacetin, ß-sitosterol, baicalin, eugenol, moslosooflavone, palmitic acid, oroxylin-A 7-O-glucuronide, and scutevulin as the potential active compounds responsible for the efficacy of HQ against both E. coli and S. aureus. The spectrum-effect relationship was utilized to reveal the orientation activities, with the results as follows: 1) The main basic-efficacy components in HQ with antibacterial, anti-inflammatory, and anticoagulant effects were P5, P8, P9, P15, P18, P19, P20; while the general basic-efficacy components were P2, P3, P6, P7, P11, P14, P21, P22, P28. 2) The main efficacy-oriented components in HQ with antibacterial effects on E. coli were P1, P12, P17, while the general efficacy-oriented compound was P10, P24, P25, P26, P27; the main efficacy-oriented in HQ with antibacterial effects on S. aureus were P14 and the general efficacy-oriented components were P1, P12, P26, P29, P30, respectively. 3) The main efficacy-oriented components with anti-inflammatory activity were P14, P24, P25, P27, and P30, while the general efficacy-oriented components were P13, P23, P26. 4) The main efficacy-oriented compounds in HQ with effects on anticoagulation were P6 and P22; these acted by prolonging APTT through the intrinsic coagulation pathway and PT through the extrinsic coagulation pathway, respectively. 5) The pharmacodynamic status classification of Scutellaria baicalensis ingredients were confirmed by nine reference compounds exemplarily. CONCLUSION: This work established a novel strategy for active compound efficacy status identification in multi-tropism Chinese herbal medicine (Scutellaria baicalensis Georgi) based on multi-indexes spectrum-effect gray correlation analysis, the method is scientific feasible and can be applied to the effective substances identification and quality control of other CHM.

Male minipuberty involves the gonad-independent activation of preoptic nNOS neurons.

BACKGROUND: The maturation of the hypothalamic-pituitary-gonadal (HPG) axis is crucial for the establishment of reproductive function. In female mice, neuronal nitric oxide synthase (nNOS) activity appears to be key for the first postnatal activation of the neural network promoting the release of gonadotropin-releasing hormone (GnRH), i.e. minipuberty. However, in males, the profile of minipuberty as well as the role of nNOS-expressing neurons remain unexplored. METHODS: nNOS-deficient and wild-type mice were studied during postnatal development. The expression of androgen (AR) and estrogen receptor alpha (ERα) as well as nNOS phosphorylation were evaluated by immunohistochemistry in nNOS neurons in the median preoptic nucleus (MePO), where most GnRH neuronal cell bodies reside, and the hormonal profile of nNOS-deficient male mice was assessed using previously established radioimmunoassay and ELISA methods. Gonadectomy and pharmacological manipulation of ERα were used to elucidate the mechanism of minipubertal nNOS activation and the maturation of the HPG axis. RESULTS: In male mice, minipubertal FSH release occurred at P23, preceding the LH surge at P30, when balanopreputial separation occurs. Progesterone and testosterone remained low during minipuberty, increasing around puberty, whereas estrogen levels were high throughout postnatal development. nNOS neurons showed a sharp increase in Ser1412 phosphorylation of nNOS at P23, a phenomenon that occurred even in the absence of the gonads. In male mice, nNOS neurons did not appear to express AR, but abundantly expressed ERα throughout postnatal development. Selective pharmacological blockade of ERα during the infantile period blunted Ser1412 phosphorylation of nNOS at P23. CONCLUSIONS: Our results show that the timing of minipuberty differs in male mice when compared to females, but as in the latter, nNOS activity in the preoptic region plays a role in this process. Additionally, akin to male non-human primates, the profile of minipuberty in male mice is shaped by sex-independent mechanisms, and possibly involves extragonadal estrogen sources.

Synthesis, and anti-inflammatory activities of gentiopicroside derivatives.

A series of 26 novel derivatives have been synthesized through structural modification of gentiopicroside, a lead COX-2 inhibitor. And their in vivo and in vitro anti-inflammatory activities have been investigated. The in vitro anti-inflammatory activities were evaluated against NO, PGE2, and IL-6 production in the mouse macrophage cell line RAW264.7 stimulated by LPS. Results showed that most compounds had good inhibitory activity. The in vivo inhibitory activities were further tested against xylene-induced mouse ear swelling. Results demonstrated that several compounds were more active than the parent compound gentiopicroside. The inhibition rate of the most active compound P23 (57.26%) was higher than positive control drug celecoxib (46.05%) at dose 0.28 mmol·kg-1. Molecular docking suggested that these compounds might bind to COX-2 and iNOS. Some of them, e.g P7, P14, P16, P21, P23, and P24, had high docking scores in accordance with their potency of the anti-inflammatory activitiy, that downregulation of the inflammatory factors, NO, PGE2, and IL-6, was possibly associated with the suppression of iNOS and COX-2. Therefore, these gentiopicroside derivatives may represent a novel class of COX-2 and iNOS inhibitors.

Antioxidant and Antimicrobial Potential, BSA and DNA Binding Properties of Some 3-Hydroxy-3-Pyrrolin-2-Ones Bearing Thenoyl Fragment.

BACKGROUND: It is known that pyrrolidinone derivates belong to a class of biologically active compounds with a broad spectrum of biological actions. Nowadays, many scientists are making an effort in the discovery of more effective ways to eliminate reactive oxygen species (ROS) that cause oxidative stress or to eliminate the harmful microorganisms from the organism in humans. Therefore, pyrrolidinones seem to be great candidates for the research of this field. METHODS: The antimicrobial activity of tested compounds was estimated by the determination of the minimal inhibitory concentration by the broth micro-dilution method against four species of bacteria and five species of fungi. The antioxidant activity was evaluated by free radical scavenging and reducing power. RESULTS: Among the tested compounds, P22 showed marked antibacterial activity on Staphylococcus aureus with a MIC value of 0.312 mg/mL. Maximum antifungal activity with MIC value 0.625 mg/mL was shown by P23 and P25 compounds against Trichophyton mentagrophytes. Tested samples showed a relatively strong scavenging activity on DPPH radical (IC50 ranged from 166.75- 727.17 µg/mL). The strongest DPPH radical scavenging activity was shown by P3 compound with an IC50 value of 166.75 µg/mL. Moreover, the tested compounds had effective reducing power. Compounds P3, P10, and P13 showed the highest reducing power than those from the other samples. Results of the interactions between DNA and P3 indicated that P3 had the affinity to displace EB from the EB-DNA complex through intercalation [Ksv = (1.4 ± 0.1) × 105 M-1], while Ka values obtained via titration of BSA with P23 or P25 [Ka = (6.2 ± 0.2) and (5.0 ± 0.2) × 105 M-1] indicate that the notable quantity of the drug can be transmitted to the cells. CONCLUSION: Achieved results indicate that our compounds are potential candidates for use as medicaments.

Correlation between sensitization to house dust mite major allergens, age, and symptoms in Japanese house dust mite allergic subjects.

Der p 23 has recently been recognized as a new house dust mite (HDM) major allergen that may be linked to the development of asthma in HDM allergic patients. This study aimed to investigate the frequency of sensitization to HDM major allergen components including Der p 23 and to examine the correlation between HDM-sensitization and AR symptom score in Japanese HDM allergic rhinitis (AR) patients without allergic asthma. Serum samples (n = 120) collected from Japanese HDM AR patients (12 to 64 years) without asthma were assessed for allergen-specific IgE (s-IgE) by ImmunoCAP (Dermatophagoides pteronyssinus (D. pteronyssinus; Der p) extract, Der p 23) or immunosolid-phase allergen chip (Der p 1, Der p 2). Japanese HDM AR patients without asthma showed a high prevalence of allergic sensitization to the HDM major allergens Der p 1 (94.2%), Der p 2 (97.5%) and Der p 23 (71.7%). No difference in the prevalence was detected for Der p 1 and Der p 2 s-IgE among three age groups. However, the prevalence of Der p 23 s-IgE was significantly higher in the younger group compared to the elderly group. No significant correlation was found between AR symptom scores and concentration of s-IgE towards Der p extract and any of the three HDM major allergens. Although the prevalence of sensitization towards D. pteronyssinus major allergens is high in Japanese AR patients without asthma, there was no correlation between allergen specific IgE including IgE towards Der p 23 and AR symptom in this population.

Ailanthus Altissima-derived Ailanthone enhances Gastric Cancer Cell Apoptosis by Inducing the Repression of Base Excision Repair by Downregulating p23 Expression.

Chemotherapy plays an irreplaceable role in the treatment of GC, but currently available chemotherapeutic drugs are not ideal. The application of medicinal plants is an important direction for new drug discovery. Through drug screening of GC organoids, we determined that ailanthone has an anticancer effect on GC cells in vitro and in vivo. We also found that AIL can induce DNA damage and apoptosis in GC cells. Further transcriptome sequencing of PDX tissue indicated that AIL inhibited the expression of XRCC1, which plays an important role in DNA damage repair, and the results were also confirmed by western blotting. In addition, we found that AIL inhibited the expression of P23 and that inhibition of P23 decreased the expression of XRCC1, indicating that AIL can regulate XRCC1 via P23. The results of coimmunoprecipitation showed that AIL can inhibit the binding of P23 and XRCC1 to HSP90. These findings indicate that AIL can induce DNA damage and apoptosis in GC cells. Meanwhile, AIL can decrease XRCC1 activity by downregulating P23 expression to inhibit DNA damage repair. The present study sheds light on the potential application of new drugs isolated from natural medicinal plants for GC therapy.

[The effect of ximedon on cholesterol metabolism and experimental atherosclerosis in rabbits]. / Vliianie ksimedona na metabolizm kholesterina i éksperimental'nyi ateroskleroz u krolikov.

The effects produced by the two pyrimidine derivatives pyridinol carbamate (parmidine) and xymedon on cholesterol metabolism and experimental atherosclerosis were comparatively studied in rabbits. The rabbits were fed either a chow containing cholesterol (200 mg/kg body weight) or the same diet also containing xymedon (30 mg/kg body weight) or pyridinol carbamate (30 mg/kg body weight). Total plasma cholesterol showed 5.5- and 4.7-fold increases in the rabbits receiving only cholesterol or cholesterol + pyridinol carbamate, respectively, as compared with that in the animals on a standard laboratory chow. In the rabbits given cholesterol+xymedon, cholesterol levels were 24% less than that in the animals taking cholesterol alone. In these animals, the aortic atherosclerotic damage index (ADI) was equal to 24.1%, which was 1.8-fold less than that in the cholesterol-fed rabbits. In the rabbits given cholesterol+pyridinol carbamate, ADI was decreased by 1.7 times, but it did not differ from that in the hypocholesterolemic rabbits. At the same time xymidone and pyridinol carbamate reduced the hepatic levels of total and esterified cholesterol. To elucidate the mechanism of action of xymedon, it was studied for effects on cholesterol metabolism in cultured rabbit hepatocytes and murine macrophage J774. Xymedon did not alter the esterification and other parameters of cholesterol metabolism in the cultured hepatocytes. It is suggested that the hypocholesterolemic effect was realized at the level of intestinal rather than hepatic cholesterol metabolic changes. The investigations made on the murine macrophage J744 showed that xymedone reduced cholesterol esterification in macrophages, evidently by inhibiting the activity of the enzyme acyl-CoA: cholesterol acyltransferase. The anti-atherosclerotic effect of xymedon seems to result from reductions in plasma cholesterol levels and cholesterol esterification in blood vascular cells.

[The comparative characteristics of the antiatherogenic effect of xymedon and parmidine]. / Sravnitel'naia kharakteristika antiaterogennogo éffekta ksimedona i parmidina.

We've studied xymedon (30 mg/kg, 12 m) and parmidin (pyridinolcarbamate 30 mg/kg, 12 m.) effects on the development of experimental cholesterol atherosclerosis on rabbits. It was demonstrated that after the use of xymedon aorta damage reduces twice. Xymedon also prevents accumulation of cholesterol and fatty degeneration of the liver. Antiatherogenic effect of xymedon resulted comparable with the effect of parmidin. As it was demonstrated in the present investigation and in the previous ones in contrast to parmidin, xymedon normalizes the lipoproteins balance owing to increase of HDL cholesterol. It also has regenerative activity on endotheliocytes and immunomodulator properties. That can be of great importance for the anti-atherosclerotic effects of the medicine.

Short Latency Somatosensory Evoked Potentials and Electroencephalography in Stroke

Median nerve somatosensory evoked potentials(SEP) and electroencephalography(E EG) were recorded in 85 patients with stroke(33 with thalamic hemorrhage, 20 with putaminal hemorrhage and 32 with cerebral infarction) to observe the origin of Nl9 and P23 wave responses in median SEP and the origin of slow waves in EEG as well as to evaluate the prognostic correlation between stroke patients and SEP and EEG findings. Nl9 and P23 were absent in 42 4% of patients with thalamic hemorrhage and 70% with putarninal hemorrhage. There was no case in which only P23 was absent in these two groups. In cerebral infarction, the most frequent finding was that both N19 and P23 were absent. P23 was absent with intact Nl9 in 2 cases with localized cortical infarction. Therefore we suggest that N19 develops in thalamus or thalamocatical pathway and P23 in the parietal cortex. There was no significant difference of EEG findings between thalamic hemorrhage and cerebral infarction. It was unlikely that slow waves on EEG is a specific finding in a localized brain lesion. The prognosis was poor in thalamic hemorrhage and cerebral infarction with loss of both Nl9 and P23 in SEP findings and in cerebral infarction with moderate to severe degree of background abnorrnalities in EEG findings. So that, SEP and EEG findings may be useful for prognostic aspect.

[The antioxidant properties of parmidine in a toxic lesion of the liver]. / Izuchenie antioksidantnykh svoistv parmidina v usloviiakh tokischeskogo porazheniia pecheni.

The experiments on rats with toxic lesions of the liver induced by tetrachlorinemethane showed that parmidine (pyridinolcarbamat) promotes the reduction of the activity of lipid peroxidation in the liver tissue homogenates and erythrocytes. The pronounced tendency towards normalization of free-radical oxidation correlated with the positive dynamics of the histological picture of the liver.

[The angioprotector and hypolipemic activity of leukomizin in experimental atherosclerosis]. / Angioprotektornaia i gipolipidemicheskaia aktivnost' leukomizina pri éksperimental'nom ateroskleroze.

A sesquiterpenic lactone leucomyzin possessing the anti-inflammatory activity significantly reduces the extensiveness of atherosclerotic lesions of the aortas of the rabbits receiving dietary cholesterol. The drug prevents an increase of permeability of the aorta and microvessels under the influence of cholesterol, bradykinin, ovalbumin and histamine. In addition to the angioprotective action, leucomyzin possesses the hypolipidemic activity in rats with experimental hyperlipidemias and (after prolonged administration) in rabbits with cholesterol atherosclerosis. By its antiatherosclerotic and angioprotective activity leucomyzin is superior to prodectine, by its hypolipidemic action it is comparable with miscleron.

[Evaluation of the therapeutic effect of hyperbaric oxygenation and anginin in peptic ulcer]. / Otsenka terapevticheskogo éffekta giperbaricheskoi oksigenatsii i anginina pri iazvennoi bolezni zheludka i dvenadtsatiperstnoi kishki.

Hyperbaric oxygenation (HBO) and pyridinolcarbamat were mainly applied in the treatment of elderly patients with peptic ulcer attended by coronary heart disease and atherosclerosis. To evaluate the condition of intracellular regulators after the treatment, a study was made of the time-course of changes in the content of cyclic nucleotides in blood plasma and gastric mucosa. The use of HBO in the treatment of peptic ulcer raised the efficacy of the multimodality therapy and accelerated the epithelization of erosive and ulcerative defects. The use of pyridinolcarbamat turned to produce a beneficial therapeutic effect and to be protective with respect to the gastric mucosa. Thus it was found desirable to apply the drug to the treatment of ulcers mainly occurring in the stomach. The changes in the content of cyclic nucleotides in blood plasma and gastric mucosa may play an important role in the realization of positive metabolic changes in gastric mucosa, induced by pyridinolcarbamat and HBO in peptic ulcer patients.

[Effect of kininotropic preparations on ulcer formation in rats]. / Vliianie kininotropnykh preparatov na iazvoobrazovanie krys.

The effects of an inhibitor of kininogenesis trasylol, its activator, cellulose sulfate, and inhibitors of kininases, captopril and D-penicillamine, as well as an antagonist of kinins, parmidine, on the development of the rat ulcers induced by histamine, absolute ethanol, reserpine and diclofenac sodium were studied. It was found that the drugs influencing metabolism and effects of kinins (except trasylol) exert differently pronounced antiulcer effects during modelling gastric ulcers by administering ethanol, reserpine and diclofenac sodium. At the same time they were inactive on the model of histamine-induced ulcers in rats.

[Growth and metastasis of Lewis' lung carcinoma in mice under the pharmacological action on metabolism and the effects of kinins]. / Rost i metastazirovanie kartsinomy legkikh L'iuis u myshei pri farmakologicheskikh vozdeistviiakh na metabolizm i éffekty kininov.

In experiments on C57 Bl mice the effects of growth and metastatic proliferation of Lewis lung carcinoma inoculated intramuscularly on metabolism and kinin activity have been studied during the application of pharmacologic agents (parmidin, cellulose sulphate, unithiol). Causing a weak inhibition of the primary tumour growth, parmidin if administered repeatedly in given doses, inhibits partially metastatic process, which is manifested in an essential decrease of the quantity, size and weight of lung metastases. Parmidin also inhibits an adverse effect of cellulose sulphate and unithiol on metastatic process. It is believed that the action of parmidin is of an indirect character and is mainly associated with the elimination of the kinin component of microcirculatory disorders in the lungs.

[Antagonism of parmidin to kinin effects]. / Izuchenie antagonizma parmidina k éffektam kininov.

Parmidin in 10(-9)-10(-5) g/ml concentrations displays specific and competitive antagonism to spasmogenic effects of bradykinin and methionyl-lysil-bradykinin on isolated segments of the guinea pig ileum, rat uteral cornua, cat jejunum and spiral strips of the rabbit aorta. Its antagonism to the spasmogenic effect of lysil-bradykinin is of both competitive and non-competitive character. In rats parmidin in 100 mg/kg dose decreased the paw edema, induced by kinins but it did not influence the similar effect of histamine and that of prostaglandin F2.

[Formalin-induced minor tremor response as an indicator of pain].

Formalin which was said to produce prolonged pain and inflammation was injected subcutaneously into the back of guinea pigs, and minor tremor pain response (MTP-response) was measured using the MT-pick up, integrator and digital volt meter. The MTP-response curve showed a biphasic pattern. Immediately after injection, the MTP-response curve showed a significant peak which lasted for about 2 min (the first phase) and subsequently dipped rapidly, and after 5 min, it began to rise slowly again and had a peak at 30 min (the second phase). Morphine (6 mg/kg, s.c.) inhibited completely the first and second phases. Levallorphan (1.2 mg/kg), however, reversed the inhibitory effect of morphine at the first phase, but not at the second phase. Aspirin (200 mg/kg, i.p.), aminopyrine (100 mg/kg, s.c.) and pentazocine (5 mg-10 mg/kg, s.c.) inhibited significantly the formalin-induced MTP-response at both phases. Pyridinol carbamate (200 mg/kg, i.p.) and hydrocortisone (25 mg/kg, i.p.) had no effect on the MTP-response at the first phase, but inhibited it at the second phase. There was a parallelism between the time course of the vascular permeability induced by formalin and that of the second phase of MTP-response. From these results, it is suggested that the first phase of MTP-response is derived from the direct effect of formalin on free nerve endings, while the second phase is derived from the inflammation. Since two kinds of pain features were differentiated in this method, the relationships with so-called "immediate pain" and "delayed pain" were discussed. Furthermore, this method can be utilized to assess pain and the action of analgesics objectively and quantitatively.

Mutagenicity of N-nitroso pyridinol carbamate with the Salmonella/mammalian microsome test.

Pyridinol carbamate was nitrosated to the N,N1-dinitroso derivative and the structure was proved by spectroscopic methods, including chemical ionization mass spectroscopy. By the Ames test, the dinitroso derivative showed a significant dose-dependent mutagenic response with Salmonella typhimurium strains TA 1535 and TA 100; the response became more pronounced in the presence of microsomes. As the dosage of N-nitroso pyridinol carbamate increased, the number of revertant colonies also increased. Pyridinol carbamate was not mutagenic.