Chemical composition and absorption characteristics of Angong Niuhuang Pill based on two-dimensional ultra-high-performance liquid chromatography-quadrupole Orbitrap high-resolution mass spectrometry.

Angong Niuhuang Pill (ANP) is a traditional Chinese medicine (TCM) formula with significant clinical efficacy in the treatment of stroke. Due to its complex composition, little attention has been directed toward the analysis of chemical composition and absorption characteristics of ANP. In this study, a reliable two-dimensional ultra-high-performance liquid chromatography (2D-UHPLC) coupled with quadrupole-Orbitrap high-resolution mass spectrometry (Q-Orbitrap HRMS) method was established to characterize the chemical constituents in ANP as well as the prototype components and metabolites absorbed in plasma, urine, feces, and brain tissues after oral administration. The prototype components were identified by a high mass accuracy (within 5 ppm) and MS/MS data based on online, local, and ANP self-built databases. The metabolites were predicted and identified using Compound Discoverer metabolic platform. A total of 154 compounds mainly including 37 flavonoids, 35 alkaioids, 19 organic acid, 19 bile acid, 32 terpenoids and 12 others were identified in this way. In addition, 60 prototype components mainly including flavonoids, alkaioids, organic acid, terpenoids and 164 metabolites were confirmed or preliminarily identified in rats. The metabolic pathways phase I reaction (hydration, reduction, oxidation, demethylation, and hydroxylation) and phase II reaction (acetylation, stearyl conjugation, and methylation) for the absorbed constituents were explored and summarized. This is the first systematic and comprehensive chemical characterization in ANP and its metabolism in vivo by 2D-UHPLC-Q-Orbitrap HRMS. This approach provides an effective strategy for the characterization of compounds and metabolites in traditional Chinese medicine formulas.

Effectiveness and safety of Angong Niuhuang pill in treatment of acute stroke: a systematic review and Meta-analysis.

OBJECTIVE: To systematically evaluate the efficacy and safety of Angong Niuhuang pill (, ANP) in the treatment of acute stroke. This can provide ideas and basis for the treatment of this disease with integrated Traditional Chinese and Western Medicine. METHODS: Randomized controlled trials (RCTs) of China National Knowledge Infrastructure Database, Wanfang Database, Chinese BioMedical Literature Database, PubMed, Embase, and the Cochrane Library were searched from the establishment to March 2022. Two researchers screened the literature and extracted the data according to inclusion and exclusion criteria. Meta-analysis was performed using RevMan 5.3 software. RESULTS: A total of 28 RCTs were included, including 2745 patients in the acute stage of stroke (1375 in the experimental group and 1370 in the control group). Meta-analysis showed that compared with conventional treatment, combined treatment with ANP could improve the effective rate of acute stroke patients [relative risk () = 1.26, 95% confidence interval () (1.21, 1.31)], Glasgow Coma Scale scores [mean difference () = 2.01, 95% (1.04, 2.98)], Mini-mental State Examination scores [ = 4.79, 95% (2.22, 7.37)], Activities of Daily Living scores [ = 15.70, 95% (14.05, 17.36)] and the Barthel index scores [ = 13.89, 95% (12.12, 15.65)], reduce National Institute of Health stroke scale scores [ = -3.90, 95% (-4.96, -2.84)] and serum brain natriuretic peptide [ = -38.50, 95% (-46.85, -30.15)]. In terms of safety, the incidence of adverse reactions showed no statistical differences between the two groups [ = 0.71, 95% (0.43, 1.15), = 0.16], and no serious adverse reactions/events were observed, indicating a good safety. CONCLUSIONS: Existing clinical research evidence shows that ANP has good efficacy and safety in the treatment of acute stroke, which can provide a basis for the treatment of integrated Traditional Chinese and Western Medicine. However, the quality of included research methodology needs to be improved, and the above conclusions need to be verified by more high-quality studies.

Characterization and quantification of chemical constituents in Angong Niuhuang Pill using ultra-high performance liquid chromatography tandem mass spectrometry.

Material basis researches and quality evaluation of traditional Chinese medicine preparations (TCMPs) face great challenges due to the complex composition and abundant unexpected interference. Angong Niuhuang Pill (ANP), one of the most famous TCMPs in China, containing 11 crude drugs has been commonly used for the treatment of febrile diseases. However, previous literatures of comprehensive chemical constituents in ANP were still limited. Herein, Ultra-high performance liquid chromatography-ion mobility-quadrupole time-of-flight mass spectrometry (UHPLC-IM-QTOF MS) method was established to effectively recognize the chemicals in Angong Niuhuang Pill (ANP). A total of 205 compounds, containing 72 confirmed with reference standards, were characterized from ANP with the application of a systematic strategy integrated polygonal mass defect filtering (MDF) and diagnostic fragment ion filtering (DFIF)/neutral loss filtering (NLF). Additionally, 68 major constituents in 8 batches of ANP samples were simultaneously determined by ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QQQ MS/MS) within 40 min. The quantitative method was validated regarding linearity, precision, repeatability, stability, and accuracy. These findings proved the established liquid chromatography-mass spectrometry (LC-MS) method was efficient and dependable for qualitative and quantitative chemical profiling of ANP. Besides, this research could provide the material basis for further pharmacological researches and quality control of ANP.

Angong Niuhuang Pill ameliorates cerebral ischemia/reperfusion injury in mice partly by restoring gut microbiota dysbiosis.

Angong Niuhuang Pill (ANP) is a famous traditional Chinese patent medicine and is used for treating ischemic or hemorrhagic stroke for centuries. However, the mechanism of action of ANP in stroke treatment has rarely been reported. With increasing evidence for a mechanistic link between acute ischemic stroke and gut microbiota alterations, this study aimed to determine the mechanism of action of ANP in treating acute ischemic stroke from the perspective of the gut microbiota. A mouse model of acute ischemic stroke by middle cerebral artery occlusion (MCAO) was established, and 16S ribosomal RNA (rRNA) gene sequencing and metabolomic analysis were performed on the cecal content samples collected from the sham, model, and ANP-treated MCAO mice. The results showed that ANP significantly ameliorated cerebral infarct volume, improved neurological deficits, and reduced histopathological injuries in the ipsilateral ischemic cortex, hippocampus, and striatum. The latter effects included inhibition of neuronal death, increased Nissl bodies, and decreased cell apoptosis. Moreover, ANP reversed gut microbiota dysbiosis by modulating the abundance of bacteria whose effects may mitigate MCAO damage, such as the phyla Bacteroidetes and Firmicutes, the families Lachnospiraceae and Prevotellaceae, and the genera Alloprevotella and Roseburia. Microbial metabolites related to inflammation and neuroprotection, such as prostaglandin I2 and uridine, were also regulated by ANP treatment. Uridine, guanosine, and inosine might be potential neuromodulators produced by the gut microbiota in the ANP-treated group. Spearman correlation analysis revealed that these metabolites were intimately related to certain genera, including Alloprevotella, Lachnoclostridium, Enterorhabdus, Roseburia, Lachnospiraceae_UCG-006, and Colidextribacter. Our results demonstrated that alleviating gut microbiota dysbiosis is one of the mechanisms by which ANP protects against ischemic stroke and suggest that targeting Alloprevotella, Lachnoclostridium, Enterorhabdus, Roseburia, Lachnospiraceae_UCG-006, and Colidextribacter might be a potential anti-stroke therapy.

Comprehensive chemical profiling of volatile constituents of Angong Niuhuang Pill in vitro and in vivo based on gas chromatography coupled with mass spectrometry.

BACKGROUND: Angong Niuhuang Pill (ANP), a renowned precious traditional Chinese medicine prescription, is extensively utilized for the clinical treatment of stroke, meningitis and encephalorrhagia in China. As a classic resuscitation-inducing aromatic prescription, ANP has been investigated for its pharmacological effects in recent years, while the volatile composition in ANP still lacks comprehensive elucidation. METHOD: To better explore the volatile constituents in ANP, a qualitative analysis method was developed based on gas chromatography coupled with mass spectrometry. Furthermore, a validated quantitative method was established to determine 21 main compounds in 8 batches of commercially available ANP samples by gas chromatography-tandem mass spectrometry. The quantitative data were successively subjected to Pearson correlation coefficient analysis. Additionally, the absorbed volatile constituents in rat plasma after single oral administration of ANP have also been characterized. RESULTS: A total of 93 volatile constituents including 29 sesquiterpenoids, 28 monoterpenoids, 13 fatty acids and their esters, 7 alkanes, 6 ketones, 3 phenols, 3 aldehydes, 2 benzoate esters, and 2 other types, were preliminarily characterized, which primarily originated from Borneolum, Moschus, Curcumae Radix, and Gardeniae Fructus. D-Borneol, isoborneol and muscone were the top three abundant ingredients (> 600 µg/g) in 8 batches of ANP samples. Subsequently, the average Pearson correlation coefficient of the contents of 21 analytes was 0.993, inferring the high batch-to-batch similarity among 8 batches. After oral administration of ANP, D-borneol, isoborneol, muscone and camphor were the main volatile constituents absorbed in the rat plasma. CONCLUSION: This research may be helpful for the comprehensive quality control study of ANP, and provide for guarantee the clinical efficacy of ANP.

Respiratory nursing care with Angong Niuhuang pill for patients with chronic obstructive pulmonary disease following cardiac surgery.

AIM: Angong Niuhuang pill (ANP) is a traditional Chinese medicine (TCM) drug widely used for treating stroke. This study aimed to investigate the effect of ANP on respiratory nursing outcomes in chronic obstructive pulmonary disease (COPD) patients following cardiac surgery. METHODS: A total of 80 COPD patients following cardiac surgery were enrolled and randomized into the control group receiving routine postoperative nursing and ANP group additionally receiving ANP treatment for 3 days (n = 40 for both group). The frequency of back percussion, time of back percussion, amount of expectoration, arterial blood gas levels were compared between groups. RESULTS: Compared to the control group, the ANP group had a significantly shorter daily mean time of back percussion at day 3 (p = .036) and day 7 (p = .014). The daily mean amount of expectoration was higher at day3 (p = .018) but lower at day 7 (p = .043) in the ANP group than in the control group. In addition, the ANP group had significantly higher hemoglobin saturation (SpO2 ) and partial pressure of oxygen (PaO2 ) but lower partial pressure of carbon dioxide (PaCO2 ) at both day 3 and day 7 than the control group (all p < .05). Furthermore, the time of postoperative aerosol inhalations (p = .041), pulmonary infection rate (p = .025) and postoperative hospital stay (p = .036) were significantly reduced in the ANP group. The ANP group had significantly lower TCM symptom scores at day 3 and day 7 after surgery. CONCLUSION: These results suggested that ANP treatment can effectively promote the postoperative recovery and respiratory nursing outcomes in COPD patients following cardiac surgery.

Adjuvant treatment with Angong Niuhuang pills in treating traumatic brain damage: a meta-analysis of randomized controlled trials.

BACKGROUND: The Angong Niuhuang pill (ANP) has been widely used in the adjuvant treatment of patients with traumatic brain injury (TBI). However, the efficacy and adverse reactions of this drug are controversial. In this study, it was aimed to evaluate the effectiveness and safety of ANP on patients with TBI by a systematic review and meta-analysis of randomized controlled trials (RCTs). METHODS: PubMed, Embase, Cochrane Library, Chinese Biomedicine Database (CBM), China National Knowledge Infrastructure (CNKI), Chinese Scientific Journals Database (VIP), and Wangfang databases were systematically searched from their establishment until June 2020. RCTs of ANP treating TBI were enrolled. Odds risk (OR) was used to assess the total effective rate and safety and mean difference (MD) and 95% confidence interval (CI) were used to assess the quantitative data. Tthe included literature's quality was evaluated by RevMan 5.3. The sensitivity and publication bias was evaluated by Stata 16.0. RESULTS: Twelve studies were identified in this systematic review, including 1,568 participants. The metaanalysis results suggested that ANP combined with routine treatment obviously improved the postoperative GCS [MD =1.97, 95% CI (1.22, 2.72), P<0.01] and GOS [OR =2.28, 95% CI (1.60, 3.22), P<0.01] of patients with TBI. ANP also increased Mg2+ concentration and decreased pulmonary infection. In addition, ANP significantly reduced NSE, gastrointestinal bleeding, and liver and kidney function damage. CONCLUSIONS: Based on limited evidence, ANP adjuvant therapy may have a clinical benefit in improving the prognosis of patients with TBI and reducing the associated complications. At the same time, more studies with larger sample sizes and high quality are required to determine the safety and effectiveness of ANP adjuvant therapy.

Protective effect and mechanism of Angong Niuhuang pill in sepsis-associated brain dysfunction of rats / 中华危重病急救医学

Objective:To observe the protective effect of Angong Niuhuang pill on brain function of rats with sepsis, explore its protective mechanism, and provide the experimental basis for clinical application of Angong Niuhuang pill in the treatment of sepsis-associated encephalopathy (SAE).Methods:Thirty male Sprague-Dawley (SD) rats were divided into sham operation group, sepsis model group and Angong Niuhuang pill group according to random number table method, with 10 rats in each group. The sepsis model was established by cecal ligation and puncture (CLP); rats in sham operation group received open and closed abdomen. The rats in the Angong Niuhuang pill group were given Angong Niuhuang pill (0.3 g/kg) by gastric irrigation daily for 3 days before CLP, and the drugs were administrated 12 hours after modeling again. After 24 hours of CLP, the neuroreflex scores were evaluated, white blood cell count (WBC), the levels of serum neuron-specific enolase (NSE) and S100β were detected. Then the brain tissue was harvested. After hematoxylin-eosin (HE) staining, the pathological changes of brain tissue were observed under the light microscope. The mRNA expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in brain tissue were detected by polymerase chain reaction.Results:Compared with the sham operation group, the total score of neuroreflex scores in the sepsis model group and the Angong Niuhuang pill group were significantly reduced (4.43±1.40, 6.57±1.90 vs. 9.40±0.84, both P < 0.05), WBC, serum NSE, S100β were significantly increased [WBC (×10 9/L): 8.07±1.32, 5.84±0.94 vs. 3.60±0.32; NSE (μg/L): 1.04±0.14, 0.61±0.07 vs. 0.16±0.04; S100β (ng/L): 255.624±30.25, 97.72±15.41 vs. 46.88±12.03, all P < 0.05], and the mRNA expressions of IL-6 and TNF-α in brain tissue were significantly increased [IL-6 mRNA (2 -ΔΔCt): 5.668±2.195, 3.605±1.014 vs. 0.997±0.329; TNF-α mRNA (2 -ΔΔCt): 18.996±0.913, 1.746±0.710 vs. 0.674±0.132, all P < 0.05]. Compared with the sepsis model group, the total score of neuroreflex scores in the Angong Niuhuang pill group was significantly increased (6.57±1.90 vs. 4.43±1.40, P < 0.05), WBC, serum NSE, S100β concentration, and the mRNA expressions of IL-6 and TNF-α in the brain were significantly reduced [WBC (×10 9/L): 5.84±0.94 vs. 8.07±1.32, NSE (μg/L): 0.61±0.07 vs. 1.04±0.14, S100β (ng/L): 97.72±15.41 vs. 255.62±30.25, IL-6 mRNA (2 -ΔΔCt): 3.605±1.014 vs. 5.668±2.195, TNF-α mRNA (2 -ΔΔCt): 1.746±0.710 vs. 18.996±0.913, all P < 0.05]. Brain histopathological observation showed that the hippocampal neurons in the sepsis model group were disordered arrangement, a large number of neuronal nuclei were contracted, and the tissue was loose with obvious edema. Compared with the sepsis model group, the Angong Niuhuang pill group had less nuclear shrinkage and tissue edema. Conclusions:The pretreatment of the Angong Niuhuang pill can improve the brain dysfunction of septic rats and reduce the expression of pro-inflammatory cytokines in the brain. It is speculated that the Angong Niuhuang pill can protect the brain function in sepsis by inhibiting the inflammatory reaction in the brain.

Angong Niuhuang Pill as adjuvant therapy for treating acute cerebral infarction and intracerebral hemorrhage: A meta-analysis of randomized controlled trials.

ETHNOPHARMACOLOGICAL RELEVANCE: Angong Niuhuang Pill (ANP) is a well-known traditional Chinese patent medicine. This meta-analysis aimed to evaluate the efficacy and safety of ANP as an adjuvant therapy in patients with acute cerebral infarction (ACI) and acute intracerebral hemorrhage (AIH). MATERIALS AND METHODS: We performed a literature search in Embase, Pubmed, Cochrane Library, CNKI, Wanfang, and VIP database from their inceptions to April 2018. Randomized controlled trials evaluating ANP as an adjuvant therapy for acute stroke were selected. Risk ratio (RR) or weighted mean difference (WMD) with their 95% confidence interval (CI) was calculated between with and without ANP therapy. RESULTS: Eighteen trials involving 1,601 patients were identified and analyzed. Meta-analysis showed that ANP plus usual treatment significantly improved the total response rate in patients with ACI (RR 1.27; 95% CI 1.14-1.41) and AIH (RR 1.26; 95% CI 1.14-1.38) compared with the usual treatment alone. Adjuvant treatment with ANP also significantly reduced the neurologic deficit score in patients with ACI (WMD -3.64; 95% CI -4.97 to - 2.31) and AIH (WMD -3.52; 95% CI -5.51 to -1.54). Moreover, ANP significantly improved the Glasgow Coma Scale in patients with ACI (WMD 1.18; 95% CI 0.79-1.56) and AIH (WMD 2.28; 95% CI 1.37-3.19). CONCLUSIONS: Adjuvant treatment with ANP appears to improve the total response rate and neurologic deficit score in patients with ACI and AIH. More well-designed trials are required due to the suboptimal methodological quality of the included trials.

Anti-Atherosclerosis Effect of Angong Niuhuang Pill via Regulating Th17/Treg Immune Balance and Inhibiting Chronic Inflammatory on ApoE-/- Mice Model of Early and Mid-Term Atherosclerosis.

Angong Niuhuang Pill (ANP) is a well-known patented Chinese medicine which is used for hundreds of years for treating the central nervous system diseases. Atherosclerosis is a poly-aetiological chronic inflammatory vascular disease. Preventing inflammation is fundamental for treating atherosclerosis in early stages. In this study, we investigated the protective effects and possible mechanisms of ANP action on a high-fat diet induced early and mid-term atherosclerosis ApoE-/- mice. The effects of ANP were compared with accepted drug simvastatin. Twelve male C57BL/6J mice were used as the control group, and 60 male ApoE-/- mice were randomly divided into five groups: Model group, Simvastatin group, Low-, Medium-, and High-dose ANP group these groups received, respectively, saline, simvastatin (3.0mg/kg), low-dose ANP (0.25 g/kg), medium-dose ANP (0.50 g/kg), and high-dose ANP (1.0 g/kg), once every other day for 10 weeks. After administration, serum biochemical indices were detected by the automatic biochemical analyzer, the concentrations of IL-6 and IL-10 in the serum were assayed by ELISA, expression levels of IL-1ß, TNF-α, MMP-2, MMP-9, CCL2, and its receptor CCR2 in the full-length aorta, and expression levels of transcription factors Foxp3, RORγt in the spleen were assayed via western blotting and RT-qPCR. Flow cytometry was used to analyze Th17 cells and Treg cells. Pathological and histological analysis was completed on aortic root. ANP decreased LDL/HDL ratio, concentrations of IL-6 while increased IL-10 in serum. Moreover, ANP down-regulated the expression levels of IL-1ß, TNF-α, MMP-2, MMP-9, CCL2, and CCR2 receptor in the full-length aorta. In addition, ANP decreased Th17 cells and expression levels of transcription factor RORγt, increased Treg cells and expression levels of transcription factor Foxp3. ANP decreased content of collagen fibers and infiltration of inflammatory cells in the aortic root. In conclusion, we demonstrated that ANP has anti-atherosclerosis effects on a high-fat diet induced ApoE-/- mice early and mid-term AS model via regulating Th17/Treg balance, inhibiting chronic inflammation, reducing plaque collagen fibers, and reducing inflammatory cells infiltration, to exert its multi-channel multi-target anti-early and mid-term AS effects.

Meta-analysis of Angong Niuhuang Pill Combined with Chemical Drug in the Treatment of Severe Craniocerebral Injury and Its Effect on Patient's Mg2+ Concentration in Peripheral Blood / 中国药房

OBJECTIVE:To evaluate clinical efficacy of Angong niuhuang pill combined with chemical drug in the treatment of severe craniocerebral injury and its effect on the concentration of Mg2+ in peripheral blood,and to provide evidence-based reference in clinic.METHODS:Retrieved from Chinese Journal Full-text Database,China Science and Technology Journal Database,China Bxdxiology Medicine disc,Wanfang Database,Chinese Clinical Trial Registry,PubMed,Excerpta Media Database,The Cochrane Library,Web of Science,Clinical Trials,and related literatures of intemet searched by Google Scholar,randomized controlled trials (RCT) about Angong niuhuang pill combined with chemical drug (trial group) vs.chemical drug (control group) in the treatment of severe craniocerebral injury and its effects on the concentration of Mg2+ in peripheral blood were collected.After literature screening,data extraction,quality evaluation with modified Jadad scale,meta-analysis of Glasgow Coma Scale (GCS) scores and Mg2+ concentration in peripheral blood were conducted by using Rev Man 5.3 statistical software after 7 d of treatment.RESULTS:A total of 6 RCTs were included,involving 773 patients.Results of meta-analysis showed that GCS [MD=2.87,95％CI (1.64,4.10),P＜0.01] and Mg2+ concentration in peripheral blood [MD=0.11,95％CI(0.06,0.16),P＜0.01] of trial group were significantly higher than those of control group,with statistical significance.CONCLUSIONS:Therapeutic efficacy of Angong niuhuang pill combined with chemical drug is better than that of chemical drug alone in the treatment of severe craniocerebral injury,can improve clinical symptom and prognosis.

Anti-atherosclerosis and cardio-protective effects of the Angong Niuhuang Pill on a high fat and vitamin D3 induced rodent model of atherosclerosis.

ETHNOPHARMACOLOGICAL RELEVANCE: The Angong Niuhuang Pill (ANP) is a well known Chinese traditional therapeutic for the treatment for diseases affecting the Central Nervous System (CNS). Components of the ANP formulation, including Bovis Calculus Sativus, Pulvis Bubali Comus Concentratus, Moschus, Margarita, Cinnabaris, Realgar, Coptidis Rhizoma, Scutellariae Radix, Gardeniae Fructus, Curcumae Radix, and Bomeolum Syntheticum, have been used for the treatment of stroke, encephalitis and emergency meningitis across Asia, especially in China for hundreds of years. OBJECTIVE: The goal of this study was to investigate the anti-atherosclerosis and cardio-protective effects of ANP administration using a rodent model of atherosclerosis induced by a high fat and vitamin D3. METHODS: Specific Pathogen-Free (SPF) 78 male SD rats were randomly divided into a control group and 5 atherosclerotic model groups. The atherosclerotic groups were divided to receive either Simvastatin (SVTT, 0.005g/kg), Low-dose ANP (0.125g/kg), Medium-dose ANP (0.25g/kg), and High-dose ANP (0.5g/kg). Following adaptive feeding for one week, atherosclerosis was induced and the atherosclerosis model was established. Experimental drugs (either simvastatin or ANP) or normal saline were administered intragastrically once daily for 9 weeks starting from the 8th week. A carotid artery ultrasound was performed at the 17th week to determine whether atherosclerosis had been induced. After the atherosclerosis model was successfully established, platelet aggregation rates, serum biochemical indices, apoptosis-related Bcl-2, Bax proteins levels in the heart were assayed. Pathological and histological analysis was completed using artery tissue from different experimental different groups to assess the effects of ANP. RESULTS: ANP significantly decreased aortic membrane thickness, the maximum platelet aggregation rates, and the ratio of low density lipoprotein cholesterol (LDL) to high density lipoprotein cholesterol (HDL). In addition, ANP significantly reduced serum contents of total cholesterol, low density lipoprotein, malondialdehyde, troponin I, high-sensitivity C-reactive protein, and lactate dehydrogenase. ANP markedly improved abnormal pathological conditions of the aorta and heart, and helped to prevent myocardial apoptosis. CONCLUSIONS: We have demonstrated that ANP has robust ant-atherosclerosis and cardio-protective effects on a high-fat and vitamin D3 - induced rodent model of atherosclerosis due to its antiplatelet aggregation, lipid regulatory, antioxidant, anti-inflammatory and anti-apoptotic properties.

An prospective observation on clinical therapeutic effect of Lund program combined with Angong Niuhuang pill for treatment of patients with severe traumatic craniocerebral injury / 中国中西医结合急救杂志

Objective To observe the therapeutic effect and prognosis of patients with severe traumatic craniocerebral injury treated by Lund program combined with Angong Niuhuang pill. Methods A prospective study was conducted. Forty-nine patients with severe traumatic craniocerebral injury were divided into two groups:combined treatment group (25 cases) and control group (24 cases). All the patients in the two groups received conventional therapy. Additionally, the combined treatment group was treated by the Lund program combined with Angong Niuhuang pill, the patients after admission into the intensive care unit (ICU), indwelling of gastric tube was immediately given (the tube inserted and retained through mouth in cases with cranial base bone fracture), one pill of Angong Niuhuang was fed nasally twice a day for consecutive 15 days. Before and after treatment, the intracranial pressure (ICP), cerebral perfusion pressure (CPP), Glasgow coma scale (GCS) score, the mean flow velocity of middle cerebral artery (MCA), and the incidence of complication, clinical efficacy and prognosis in the two groups were observed. Results There were no statistically significant differences in the levels of ICP, CPP, GCS score and the average flow velocity of MCA before treatment and the first day after treatment between the two groups. Beginning from 3 days after treatment, the degrees of improvement in the above indexes in the combination treatment group became much better than those in the control group [ICP (mmHg, 1 mmHg=0.133 kPa):16.78±4.50 vs. 18.80±5.67, CPP (mmHg):71.35±1.63 vs. 58.11±1.47, GCS score:9.53±0.38 vs. 8.23±0.44, the mean blood flow velocity (cm/s):152.23±3.33 vs. 178.86±5.23, all P0.05). The rate of good therapeutic effect in combination treatment group was obviously higher than that of the control group [72.0% (18/25) vs. 33.3% (8/24), P < 0.05], while the mortality was markedly lowered [8.0% (2/25) vs. 33.3% (8/24), P < 0.05]. Conclusions The combination of Lund program and Angong Niuhuang pills for treatment of patients with severe traumatic craniocerebral injury can reduce the degree of increase of ICP at early stage, ameliorate cerebral edema and its development, elevate CPP and improve cerebral ischemia and hypoxia. In the aspects of controlling hyperpyrexia, decreasing limbs twitches and the incidence of hernia, elevating the quality of patients' life, reducing mortality and improving prognosis, the therapy has relatively good therapeutic effects.

Type and quality study of Calculus Bovis in commercial Angong Niuhuang Pills / 中国药学杂志

OBJECTIVE: To study the type and quality of Calculus Bovis in commercial Angong Niuhuang Pills (traditional Chinese medicines) and provide technical support to fight against counterfeits. METHODS: Specific and effective components of Calculus Bovis were studied. The type of Calculus Bovis in commercial Angong Niuhuang Pills was identified according to the differences in components and content. RESULTS: Illegal use of artificial Calculus Bovis or bilirubin was found to be a serious problem. The type and quality of Calculus Bovis in Angong Niuhuang Pills differed greatly. CONCLUSION: Calculus Bovis adulteration in Angong Niuhuang Pills is a very serious problem which needs urgent supervision and regulation to assure safe medication.

Anti-proliferation of Angong Niuhuang pill on tumor cells via inducement of apoptosis and down-regulation of mitochondrial membrane potential / 中国药理学与毒理学杂志

OBJECTIVE To validate the anticancer effect of Angong Niuhuang pill (AGNH) and pinpoint associated molecular mechanisms using human cancer cells.METHODS Human MGC-803 gastric carcinoma and human BEL-7402 hepatocarcinoma cells were incubated with AGNH 9,30,90,300 and 900 mg·L-1 for 24,48and 72 h,respectively.Cell viability was detected with 3-(4,5-dimethylthiazol-2-yl) -5-( 3-carboxymethoxyphe-nyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and colony formation assay.Apoptosis was measured with flow cytometry and Hoechst 33258/PI staining.Change in mitochondrial membrane potential (△qψ) was detected by spectrofluorophotometer.RESULTS AGNH inhibited MGC-803 cell proliferation ( for 48 h,r =0.996,P =0.002; for72 h,r=0.756,P=0.024 ) and BEL-7402 cells (for 48 h,r =0.732,P=0.030; for72 h,r=0.702,P =0.037) in a concentration-dependent manner,as showed by MTS assay.AGNH inhibited colony formation on MGC-803 cells (r =0.914,P =0.011 ) and BEL-7402 cells (r =0.871,P =0.024) in a concentration-dependent manner for 24 h.Hoechst 33258/PI staining and flow cytometry assay showed that AGNH 900 mg·L-1 for 24 h induced apoptosis of MGC-803 and BEL-7402 cells,and the apoptosis rate was 27.2％ and 19.7％,respectively.Compared with normal control group,AGNH 900 mg·L-1 for 3 min decreased the mitochondrial membrane potential of MGC-803 and BEL-7402 cells to 15.9％ and 15.0％ of control group.CONCLUSION AGNH inhibits proliferation of human cancer cells.Apoptosis and depolarization of mitochondrial transmembrane potential are probablly its mechanism.

Effect of Angong Niuhuang Pill and the heavy metal constituents on EcoG of brain damage in rats caused by LPS / 中成药

AIM:To evaluate the actions of cinnabaris and realgar in arousing brain and opening the orifices of Angong Niuhuang Pill,and to provide experimental basis for the rationality of Angong Niuhuang Pill buildup. METHODS: The brain damage model was made by LPS injection through caudal vein.With quantitative electroencephalogram of medicine methods,effects of Angong Niuhuang Pill,Angong Niuhuang Pill without cinnabaris and realgar,cinnabaris,realgar on the power and relative power of delta and beta waves of electrocorticogram(ECoG) were observed in the brain damage rats caused by LPS. RESULTS: LPS could increase the power and the relative power of the delta waves(0.5-3 Hz),and reduce the power and the relative power of the beta waves(14-30 Hz).All medicines could reduce the power and the relative power of the delta;Angong Niuhuang Pill,cinnabaris and realgar,realgar could increase the power and the relative power of the beta,cinnabaris could increase the relative power of the beta(14-30 Hz),and Angong Niuhuang Pill without cinnabaris and realgar could not. CONCLUSION: Angong Niuhuang Pill has obvious activation on ECoG,and same as cinnabaris and realgar.The results of the present work allow us to put forward the hypothesis that cinnabaris and realgar are most likely one of the important material basis in arousing brain and opening the orifices of Angong Niuhuang Pill.