Molecular targets and mechanisms involved in the action of Banxia Shumi decoction in insomnia treatment.

Insomnia is a common sleep-wake rhythm disorder, which is closely associated with the occurrence of many serious diseases. Recent researches suggest that circadian rhythms play an important role in regulating sleep duration and sleep quality. Banxia Shumi decoction (BSXM) is a well-known Chinese formula used to treat insomnia in China. However, the overall molecular mechanism behind this therapeutic effect has not yet been fully elucidated. This study aimed to identify the molecular targets and mechanisms involved in the action of BSXM during the treatment of insomnia. Using network pharmacology and molecular docking methods, we investigated the molecular targets and underlying mechanisms of action of BSXM in insomnia therapy. We identified 8 active compounds from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and the traditional Chinese medicine integrative database that corresponded to 26 target genes involved in insomnia treatment. The compound-differentially expressed genes of the BXSM network indicated that cavidine and gondoic acid could potentially become key components of drugs used for insomnia treatment. Further analysis revealed that GSK3B, MAPK14, IGF1R, CCL5, and BCL2L11 were core targets significantly associated with the circadian clock. Pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes revealed that epidermal growth factor receptor tyrosine kinase inhibitor resistance was the most prominently enriched pathway for BSXM in the insomnia treatment. The forkhead box O signaling pathway was also found to be significantly enriched. These targets were validated using the Gene Expression Omnibus dataset. Molecular docking studies were performed to confirm the binding of cavidine and gondoic acid to the identified core targets. To our knowledge, our study confirmed for the first time that the multi-component, multi-target, and multi-pathway characteristics of BXSM may be the potential mechanism for treating insomnia with respect to the circadian clock gene. The results of this study provided theoretical guidance for researchers to further explore its mechanism of action.

[Effect of electroacupuncture on cognitive functions, hippocampal HIF-1α and neurons apoptosis in postoperative cognitive dysfunction rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on learning and memory ability, hippocampal hypoxia inducible factor-1α(HIF-1α) and apoptosis in postoperative cognitive dysfunction(POCD) rats, and to investigate its mechanism underlying improvement of POCD. METHODS: A total of 90 aged male SD rats were randomized into a sham-operation group, a model group and an EA group, 30 rats in each group, which were further divided into 3 time-point subgroups (1, 3 and 7 days after intevention, 10 rats in each subgroup). In the model group and the EA group, left hepatectomy was adopted to establish the model of POCD. In the sham-operation group, the skin was sectioned and no hepatectomy was operated. In the EA group, EA was applied at "Siguan" ["Hegu" (LI 4) and "Taichong" (LR 3)] with dilatational wave, 2 Hz/100 Hz in frequency, 1 mA in intensity, 20 min each time, once a day. Morris water maze test was adopted to observe the cognitive functions. Real-time PCR and Western blot were used to measure the hippocampal level of HIF-1α. TUNEL method was used to evaluate the hippocampal level of neurons apoptosis. Double immunofluorescence labeling was used to detect the colocalization of HIF-1α and apoptosis in the EA group. RESULTS: Compared with the sham-operation group, the escape latency was prolonged and the frequency of platform leaping was reduced in the model group (P<0.05) after 1, 3, 7 days of intervention. Compared with the model group,the escape latency was shortened and the frequency of platform leaping was increased in the EA group (P<0.05) after 1, 3, 7 days of intervention. After 3 days of intervention, compared with the sham-operation group, the expressions of HIF-1α mRNA and protein, the level of apoptosis were increased in the model group (P<0.05); compared with the model group,the expressions of HIF-1α mRNA and protein, the level of apoptosis were decreased in the EA group (P<0.05). The colocalization of HIF-1α and apoptosis was observed in same cells in the EA group. CONCLUSION: Electroacupuncture improves cognitive functions in postoperative cognitive dysfunction rats, which may be related to its effect in down-regulating the expression of hippocampal HIF-1α and inhibiting the neurons apoptosis.

[Optimization of method for determination of Forsythia Fructus and discussion on the establishment of quality control standard for green (old) Forsythia Fructus].

In order to obtain the optimum method for content determination of Forsythia Fructus (FF), a variety methods for the sample preparation of FF were evaluated by the content determination methods of Chinese Pharmacopoeia. And an optimum method was screened and as follows: 30 times with 70% ethanol solution in ultrasonic extractor for half an hour. The method can achieve the best effect of simultaneously extracting forsythoside A and forsythin. Then, a HPLC method for simultaneous determination of forsythoside A and forsythin was established by methodology. The HPLC chromatographic conditions: the mobile phase consisted of acetonitrile (A)-0.4% acetic acid solution (B) with gradient elution [0-33 min，15%A，33-43 min，15%-25%A，43-60 min，25% A] was at the flow rate of 1 mL·min⁻¹, the column temperature was 25 °C, and the detection wavelength was 330 and 277 nm. Moreover, the contents of forsythoside A and forsythin for 10 Green Forsythia Fructus (GF) and 5 Old Forsythia Fructus (OF) were determined by this method and Chinese Pharmacopoeia. The result not only displayed that the established method is effective, rapid, and simple, but also showed that the contents of forsythoside A and forsythin for GF and OF were significantly different. Which implied that the forsythoside A and forsythin limit standard for GF and OF should be controled by different values. This studies provide an important basis for the establishment of the content determination of FF and the quality control standard for GF and OF.

[Role of P38-MAPK signal transduction pathway and effect of panax notoginoside in rats with hypoxic hypercapnia pulmonary hypertension].

OBJECTIVE: To investigate the role and significance of P38-MAPK in the pathological process of hypoxic hypercapnia pulmonary hypertension in rats, and the protection of panax notoginoside (PNS). METHODS: (1) To set up rat pathological model of hypoxic hypercapnia pulmonary hypertension: seventy two male SD rats (200 280 g) were randomly divided into six groups (n = 12), which were normal group (N group), hypoxic hypercapnia for 3-day group (H3d), hypoxic hypercapnia for 1-week group(H1w), hypoxic hypercapnia for 2-week group (H2w), hypoxic hypercapnia for 4-week group (H4w) and PNS-injected group (Hp). The rats of PNS -injected group were injected PNS before being placed in the chamber (50 mg/(kg x d), ip), and other groups were injected normal sodium (2 ml/kg, ip). (2) The shapes of pulmonary artery were detected by HE staining. (3) Western blot was used to study the protein expression of p38-MAPK. The expression of p38-MAPK in lung tissue and pulmonary blood vessel was investigated by immunohistochemistry. RESULTS: (1) The ratio of vessel wall area/total area (WA/ TA) in H1w, H2w, H4w and Hp group was higher than that of N group (P < 0.05), but that of H3d group did not change obviously (P > 0. 05 vs N group). The ratio of WA/TA in Hp group was obviously lower than that of H4w, group (P < 0.05). (2) The levels of P-p38 protein was markedly ascended in H3d group (0.225 +/- 0.071) compared with N group (0.012 +/- 0.006), and expression of P-p38 protein was significantly positive in H1w, H2w, H4w groups. (P < 0.05). (3) As P-p38 protein in pulmonary arterial tunica intima and tunica media, sterile expression in N group (0.099 +/- 0.015) and H3d group (0.107 +/- 0.013) contrasted to H4w group (0.124 +/- 0.025, P < 0.05), then tended to rise in H2w, H4w group (P < 0.05). (4) In pulmonary tissue, the levels of P-p38 protein in PNS-injected group were lower 53.02% (P < 0.05) than those in H4w group. In pulmonary arterial tunica intima and tunica media the levels of P-p38 protein in PNS-injected group were lower 87.33% (P < 0.05) than those in H4w group. CONCLUSION: p38-MAPK as a signal transduction may play an important role in the development of hypoxia induced pulmonary hypertension. The effect of PNS on reducing pulmonary hypertension and improving pulmonary vascular wall remodeling may be related to its inhibiting expression of p38 MAPK.

Anti-inflammatory activity of myricetin isolated from Myrica rubra Sieb. et Zucc. leaves.

MYRICA RUBRA Sieb. et Zucc. leaves are commonly used in folk medicine to treat inflammatory disorders in China. Present studies on the anti-inflammatory effect of myricetin from MYRICA RUBRA Sieb. et Zucc. leaves was evaluated with various IN VIVO models of both acute and chronic inflammations such as xylene-induced ear edema, acetic acid-induced vascular permeability, carrageenan-induced paw edema, leukocyte migration assay, and cotton pellet granuloma models. Myricetin showed a significant inhibition on ear edema and hind paw edema caused by xylene and carrageenan, respectively. Furthermore, it also inhibited the increase in capillary permeability induced by the production of acetic acid in the human body. Myricetin significantly decreased the serum levels of MDA and, in turn, increased the serum levels of SOD in the carrageenan-induced paw edema model. Concurrently, myricetin also significantly decreased leukocyte count. During chronic inflammation, myricetin inhibited the formation of granuloma tissue. These results, collectively, demonstrate that myricetin possesses a potent anti-inflammatory function on acute and chronic inflammation. Its anti-inflammatory mechanisms are probably associated with the inhibition of antioxidant activity. These results also support the claims of traditional Chinese medicine practitioners about the use of MYRICA RUBRA Sieb. et Zucc. leaves in the treatment of inflammatory diseases.

Analgesic activity of myricetin isolated from Myrica rubra Sieb. et Zucc. leaves.

Myrica rubra Sieb. et Zucc. leaves are commonly used as an astringent, antidiarrheic, and analgesics in folk medicine in China. In the present study, the analgesic activity of myricetin, a major compound in Myrica rubra Sieb. et Zucc. leaves was evaluated in vivo. The analgesic effect of myricetin was tested by a serial of models, such as acetic acid-induced writhing response, formalin-induced paw licking and hot plate test. The sedative activity was evaluated by pentobarbital-induced sleep time. Platelet aggregation induced by collagen and arachidonic acid was also performed in vitro. Myricetin showed a significant inhibition on chemical nociceptive models such as the acetic acid-induced writhing response and the licking time on the late phase in the formalin test in a dose-dependent manner, but did not manifest a signicant effect in hot plate test. Myricetin was also not able to increase the sleeping time induced by pentobarbital, which further indicated that the analgesic effect of myricetin was unrelated to sedation. In addition, myricetin inhibited the content of PGE2 in the peritoneal fluid and platelet aggregation induced by collagen and arachidonic acid in vitro. These results collectively demonstrated that myricetin possessed potent analgesic activity, which was related with peripheral analgesia, but, not with the opioid system. Myricetin may be a potent COX-1 inhibitor with anti-platelet activity.

[Influence of traditional Chinese compound recipes with different efficacy on body weight, tumor weight and immune function in H22 cancer-bearing mice].

OBJECTIVE: To compare the influence of traditional Chinese compound recipes (TCCRs) with different efficacy on body weight, tumor weight and immune function in H22 cancer-bearing mice. METHODS: H(22) cancer-bearing mice were chosen to observe the effects of TCCRs with different efficacy on tumor growth inhibition and detect the proliferation function of T lymphocytes, the activity of natural killer (NK) cells, the changes of T lymphocytes and the content of interferon-gamma (IFN-gamma)and interleukin-4 (IL-4). RESULTS: Tumor weight of H(22) cancer-bearing mice in Yidu Gongdu Recipe (YDGDR, a compound traditional Chinese herbal medicine using poison as an antidote for poison)-treated group was obviously lighter than that in the other TCCR-treated groups and the tumor inhibition rate in YDGDR-treated group was 65.76% (P<0.01). The tumor inhibition rates in other TCCR-treated groups were ranged from 10.1% to 17.1% . Body weight of mice in YDGDR-treated group was obviously decreased and depilation was observed at the same time. Pelage of mice in Fuzheng Peiben Recipe (FZPBR, a compound traditional Chinese herbal medicine for supporting the healthy energy)-treated group grew well, and behavior of the mice was active. Stimulation index (SI) of T lymphocyte transformation in YDGDR-treated group was obviously increased (SI=4.34, P<0.01), which showed the proliferation function of T lymphocyte was very strong. The SI of T lymphocyte transformation in the other groups was less than three, which showed the proliferation function of T lymphocytes was not significant. Compared with normal saline (NS)-treated group, percentages of NK cells in Qinre Jiedu Recipe (QRJDR, a compound traditional Chinese herbal medicine for clearing away heat and toxic substances)-treated, Huxue Huayu Recipe (HXHYR, a compound traditional Chinese herbal medicine for activating blood circulation to dissipate blood stasis)-treated and YDGDR-treated groups were obviously increased and 5.05, 4.07 and 5.17 times more than the NS-treated group, respectively (P<0.01). The activity of NK cells wasn't increased in the FZPBR-treated and HXHYR-treated groups. The production of IFN-gamma induced by T cells in YDGDR-treated group was obviously raised (P<0.05), and the production of IL-4 induced by T cells in QRJDR-treated, HXHYR-treated, Huatan Sanjie Recipe (a compound traditional Chinese herbal medicine for eliminating phlegm and resolving masses)-treated and YDGDR-treated groups was also raised obviously (P<0.01). CONCLUSION: YDGDR has a good effect of inhibiting tumor growth and can reinforce cellular and humoral immune function in tumor-bearing mice. FZPBR can strengthen the body.

Characteristics of and strategies for patients with severe burn-blast combined injury.

BACKGROUND: Severe burn-blast combined injury is a great challenge to medical teams for its high mortality. The aim of this study was to elucidate the clinical characteristics of the injury and to present our clinical experiences on the treatment of such cases. METHODS: Five patients with severe burn-blast combined injuries were admitted to our hospital 77 hours post-injury on June 7, 2005. The burn extent ranged from 80% to 97% (89.6% +/- 7.2%) of TBSA (full-thickness burns 75% - 92% (83.4% +/- 7.3%)). All the patients were diagnosed as having blast injury and moderate or severe inhalation injury. Functions of the heart, liver, kidney, lung, pancreas and coagulation were observed. Autopsy samples of the heart, liver, and lungs were taken from the deceased. Comprehensive measures were taken during the treatment, including protection of organ dys function, use of antibiotics, early anticoagulant treatment, early closure of burn wounds, etc. All the data were analyzed statistically with t test. RESULTS: One patient died of septic shock 23 hours after admission (four days after injury), the others survived. Dysfunction of the heart, liver, lungs, pancreas, and coagulation were found in all the patients on admission, and the functions were ameliorated after appropriate treatments. CONCLUSIONS: Burn-blast combined injury may cause multiple organ dysfunctions, especially coagulopathy. Proper judgment of patients' condition, energetic anticoagulant treatment, early closure of burn wounds, rational use of antibiotics, nutritional support, intensive insulin treatment, timely and effective support and protection of organ function are the most important contributory factors in successful treatment of burn-blast combined injuries.

[Progress of taxonomic study on Fritillaria (Liliaceae) medicinal plant].

In this review, we described the taxonomic study of the Fritillaria medicinal plant in the recent years. The taxonomic study of the Fritillaria medicinal plant was carried out from three main aspects: the traditional morphological character, the characteristic constituents of the plant and genotyping and species identification of Fritillaria by DNA chips. By comparison, we concluded that the DNA chip technology can provide a rapid, high throughput for genotyping and quality assurance of the plant species verification. It is the most prosperous method of species identification of the plant.

[Identification of Chinese medicine material of Fritillaria by themogravimetric/diffrential thermal analysis].

OBJECTIVE: To provide a quick and simple method to identify different Chinese medicine material of Fritillaria. METHOD: The thermograms and differential thermograms of nine Fritillaria powders were obtained by thermal analyzer. RESULT: By analyzing the thermograms of nine Fritillaria powders, we concluded that the thermal stability of nine Fritillaria powders was much different each other due to the different geography origin. The thermal stability of F. hupehensis was highest among nine Fritillaria, while F. ussurensis was the lowest. The different Fritillaria showed their own DTA spectra respectively. CONCLUSION: According to the differences in the thermal properties of nine Fritillaria powders, the origins of Fritillaria could be easily identfied.

[Tumor inhibiting and immunoloregulation effects of Mylabris Mixture on H22 cancer-bearing mice].

OBJECTIVE: To investigate the mechanisms of tumor inhibiting and immunoloregulation of Mylabris Mixture on H22 cancer-bearing mice. METHODS: H22 cancer-bearing mice were chosen to observe the effects of tumor inhibiting and detect the proliferation function of T lymphocytes, the toxicity function of NK cells, the changes of T lymphocytes and the contents of interferon-gamma and interleukin-4. RESULTS: Mylabris Mixture could obviously inhibit the growth of H22 cancer in mice, and the tumor inhibition rat was 65.76%. The stimulation index of T lymphocyte transformation and percentage of NK cells in Mylabris Mixture-treated group were obviously higher than those in the normal control group. The subpopulation proportion of T lymphocytes in Mylabris Mixture-treated group was changed more than the normal control group. The production of interferon-gamma and interleukin-4 by T lymphocytes obviously increased in Mylabris Mixture-treated group (P<0.05, P<0.001). CONCLUSION: Mylabris Mixture has the effect of inhibiting the growth of tumor constitution, and regulating immunological function on mice with tumor. Its mechanisms include the reinforcement of T lymphocyte immune function, NK cell killing function and humoral immune function.

[Identification of Fritillaria herbal drugs by X-ray diffraction of starch grains].

OBJECTIVE: To provide a simple method to identify different of herbal drugs of Fritillaria. METHOD: The X-ray diffraction spectra of five Fritillaria powders were obtained by X-ray diffractometry (XRD). RESULT: By analyzing the diffraction peaks of starch over the 2theta diffraction angle in the range of 10 degrees-30 degrees, we concluded that the crystal type of starch in F. thunbergii, F. ussurensis, F. pallidifloca and F. hupehensis was B-type. However, the crystal type of starch in F. cirrhosa was CB-type. The degree of crystallinity of starch was 42.1%, 43.6%, 37.7%, 35.9% and 30.8%, corresponding to F. thunbergii, F. ussurensis, F. pallidiflcoca,F. cirrhosa and F. hupehensis. CONCLUSION: Based on the differences of crystallinity of starch and X-ray diffraction spectra of Fritillaria powders, different species of Fritillaria can be identified.

[Determination of trace zinc, manganese, cadmium and lead in aloe by microwave-digestion atomic absorption spectrometry].

A microwave digestion procedure was developed for the determination of trace Zn, Mn, Cd and Pb in aloe-leaf cuticle and aloe-leaf gelatin, using the obturated vessel microwave digestion system with a pressure controlling part, and the amounts of these trace metallic elements were determined by atomic absorption spectrometry after microwave digestion. The effects of the composition of digestion solution, the ratio of the sample to digestion solution, and the digestion time were studied. It is satisfactory to apply the microwave digestion procedure to the determination of Zn, Mn, Cd and Pb under the optimized condition with the recovery of 95.0% to 110.0% and RSD of 0.3% to 6.2%. The results show that this method is rapid and simple with low environmental contamination and complete digestion of the sample.