Xiaojindan Extract Modulated Macrophage Polarization by Targeting PI3K/Akt Pathway / 中国实验方剂学杂志

ObjectiveTo explore the effect and mechanism of Xiaojindan extract （XJD） on macrophage polarization. MethodLipopolysaccharide （LPS） and interleukin-4 （IL-4） were used to induce M1 and M2 polarization of RAW264.7 cells. The influence of 10-80 mg·L-1 XJD on cell proliferation was detected by Cell Counting Kit-8 （CCK-8） assay. Nitric oxide （NO） and interleukin-6 （IL-6） release was explored by Griess assay and enzyme-linked immunosorbent assay （ELISA）， respectively. The mRNA expression of M1 and M2 macrophage markers was measured by real-time quantitative polymerase chain reaction （Real-time PCR）， and the CD206+ expression was determined by flow cytometry. The activation of phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） pathway was analyzed by western blot. Result10-80 mg·L-1 XJD showed no marked cytotoxicity in LPS （0.5 mg·L-1）- or IL-4 （20 μg·L-1）-induced RAW264.7 cells. Compared with the control group， LPS significantly promoted the expression of M1 macrophage markers （P<0.01）， including increased NO and IL-6 release （P<0.01） and upregulated mRNA expression of interleukin-1β （IL-1β）， inducible nitric oxide synthase （iNOS）， cyclooxygenase-2 （COX-2） and tumor necrosis factor-α （TNF-α） （P<0.01）. Compared with LPS-induced group， 20-80 mg·L-1 XJD decreased the release of NO and IL-6 in a dose-dependent manner （P<0.01）， and similarly 10-80 mg·L-1 XJD suppressed the mRNA expression of IL-1β， iNOS， COX-2 and TNF-α （P<0.01）. Compared with the control group， IL-4 obviously increased the expression of M2 macrophage markers （P<0.01）， including increased CD206+ cell population and upregulated mRNA expression of arginine-1 （Arg-1）， interleukin-10 （IL-10）， interleukin-13 （IL-13） and transforming growth factor-β1 （TGF-β1）. Compared with IL-4-induced group， 10-80 mg·L-1 XJD dose-dependently decreased CD206+ cell population （P<0.01） and inhibited the mRNA expression of Arg-1， IL-10， IL-13 and TGF-β1 （P<0.01）. Western blot showed that XJD significantly downregulated the activation of PI3K/Akt pathway as compared to LPS- and IL-4-induced groups （P<0.05， P<0.01）. ConclusionXJD significantly inhibited the macrophage polarization in the LPS- and IL-4-induced RAW264.7 cells by targeting PI3K/Akt pathway.

Inhibitory Effect and Mechanism of Jingulian Extract on LPS-induced RAW264.7 Cell Inflammatory Response Based on PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

Objective:To explore the inhibitory effect and mechanism of Jingulian extract （JGL） on inflammation. Method:The following groups were set up in this study： a control group （10% fetal bovine serum）， a lipopolysaccharide （LPS） model group （0.5 mg·L^-1）， and JGL groups （10， 20， 40， 60， 80， 120， 160， 200， 250， 300 mg·L^-1 + 0.5 mg·L^-1LPS）. The RAW264.7 cells were cultured for 24 hours. Cell proliferation was detected by cell counting kit-8 （CCK-8） assay. Nitric oxide （NO） release was detected by Griess assay. The release of cytokines interleukin （IL）-1<italic>β</italic>， IL-6， IL-10， and tumor necrosis factor （TNF）-<italic>α</italic> was determined by enzyme linked immunosorbent assay （ELISA）. The expression of inducible nitric oxide synthase （iNOS） and intraprostaglandin peroxidase synthase 2 （PTGS2）/cyclooxygenase-2 （COX-2） was measured by real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） and the activation of key proteins in the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway by Western blot. Result:Compared with the control group， LPS （0.5 mg·L^-1）could promote the proliferation of RAW264.7 cells after stimulation for 24 hours （<italic>P</italic><0.01）. Compared with the model group， JGL had no significant effect on cell proliferation. Compared with the control group， LPS （0.5 mg·L^-1）increased the release of NO， IL-1<italic>β</italic>， IL-6， IL-10， and TNF-<italic>α</italic> （<italic>P</italic><0.01）. Compared with the model group， JGL （20-300 mg·L^-1）inhibited the release of NO in a dose-dependent manner after stimulation for 24 hours （<italic>P</italic><0.05） and reduced IL-1<italic>β</italic>， IL-6， and IL-10 （<italic>P</italic><0.05， <italic>P</italic><0.01）， but no obvious inhibition on the release of TNF-<italic>α</italic> was observed. LPS （0.5 mg·L^-1） could induce the expression of iNOS and PTGS2/COX-2 genes as compared with the control group （<italic>P</italic><0.05， <italic>P</italic><0.01）. JGL could down-regulate the mRNA expression of iNOS and PTGS2/COX-2 genes as compared with the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. LPS （0.5 mg·L^-1） could activate the PI3K/Akt pathway （<italic>P</italic><0.01） as compared with the control group， while JGL （10， 20， 40， and 80 mg·L^-1） decreased the expression of PI3K-p110， p-p85， and p-Akt （<italic>P</italic><0.01）， and inhibited the activation of PI3K/Akt pathway. Conclusion:JGL extract could significantly inhibit the inflammatory response and activation of the PI3K/Akt pathway induced by LPS in RAW264.7 cells. The anti-inflammatory effect was related to the inhibition of the PI3K/Akt pathway.

Surgical treatment for type Stanford A aortic dissection with Kommerell's diverticulum / 中华外科杂志

To examine the outcomes of surgical treatment in patients of type Stanford A aortic dissection with Kommerell's diverticulum. From January 2009 to August 2017, patients of type Stanford A aortic dissection with Kommerell's diverticulum who underwent the Sun procedure were enrolled. Patient demographic, preoperative, intraoperative, early morbidity and mortality data were collected from medical and electronic patient records. Clinical follow-up data, including late morbidity and mortality, were obtained by telephone interview with the patient. A total of 13 patients (11 males and 2 females; mean age 47 years) were included. The mean maximum diameter of Kommerell's diverticulum was (21.8±7.7) mm. The Kommerell's diverticulum was repaired by direct suture of the orifice in 3 patients, ligation of the aberrant right subclavian artery in 9 patients, and suture and ligation in 1 patient, respectively. No perioperative death occurred. One patient underwent a reexploration for bleeding. There were 2 late deaths: unknown reason in 1 patient and septic shock secondary to renal abscess in 1 patient. Reintervention was performed in one patient for a persistent type Ⅱ endoleak. The Sun procedure with femoral artery cannulation for cardiopulmonary bypass, unilateral carotid artery cannulation for selective cerebral perfusion and ligation of aberrant right subclavian artery on the right side of the trachea is an appropriate therapeutic strategy for patients of type Stanford A aortic dissection with Kommerell's diverticulum.

Relationship between pre-clinical toxicity and efficacy of traditional Chinese medicine / 中国中药杂志

Based on the different category of syndromes between traditional Chinese medicine(TCM) and Western medicine, the relationship between the efficacy and non-clinical toxicity of the TCM was analyzed. If TCM preparations have the same pharmacological targets to treat disease with modern medicine or the Chinese herbal preparations treat the diseases with its toxicity, their toxicity often exhibits the amplification and extension of activity; on the other hand, if TCM preparations have overlapped pharmacological targets but not completely the same with modern medicine, or if they have totally different pharmacological targets, the toxicity of TCM could not be inferred by pharmacological activity. With the great progress in extraction and separation technique for active parts of TCM as well as the application of some novel technique and excipients, some toxicity may be from the reactions unrelated with the pharmacological activity. In conclusion, better design and quality control could be obtained by understanding the relationship between pharmacological and toxicological study for the investigation of new traditional medicine.

Study on main pharmacodynamics and underlying mechanisms of 999 Ganmaoling / 中国中药杂志

To observe synergistic effects of 999 Ganmaoling (GML) and its Chinese/Western materia medica (CMM and WMM) on pharmacodynamic action and to study underlying mechanisms, their anti-inflammatory, antipyretic effects were compared by assaying the increased capillary permeability induced by glacial acetic acid in mice, ear swelling induced by Xylene in mice, non-specific pleurisy induced by carrageenan in rats, and yeast induced fever in rats. Crystal violet (CV) and microbial activity (XTT) assay were used to evaluate the inhibition of GML and its CMM and WMM on KPN biofilm formation, and scanning electron microscopy (SEM) was applied for observing KPN biofilm morphology changes. The results showed that compared with control group, GML could reduce exudation amount of Evans-Blue and the degree of Ear swelling significantly, and CMM and WMM have no significant effects. The concentration of TNF-α and IL-1β of rat pleural effusion in GML, CMM and WMM group decreased significantly. The concentration of TNF-α, IL-1β and IL-8 in GML group, TNF-α, IL-8 in WMM group and IL-8 in CMM in rats serum decreased significantly. The body temperature in rats decreased significantly in GML and WMM group after 4-8 h of administration. CMM group showed no significant difference in rat body temperature compare with control. Compared with control group, GML (55-13.75 g•L⁻¹) could inhibit KPN biofilm formation and reduce number of viable cells in the KPN biofilm. CMM (45-22.5 g•L⁻¹) and WMM (10 g•L⁻¹) could also inhibit KPN biofilm formation and reduce number of viable cells (P<0.01). Result of SEM also showed that GML (55 g•L⁻¹) and its CMM (45 g•L⁻¹) and WMM (10 g•L⁻¹) could interfere the bacterial arrangement of KPN biofilm and extracellular matrix. GML and its CMM & WMM could inhibit the formation of KPN biofilm, CMM & WMM in GML showed synergism and complementation in inhibit KPN biofilm. Results showed that GML had obvious anti-inflammatory and antipyretic effects and could destruct KPN mature biofilm. WMM and CMM showed obvious synergistic effect against inflammation and inhibition of KPN biofilm formation and reduction of number of viable cells but no same effects against fever.

Study on characteristic spectrum of ingredients from different species Cordyceps and its anti-fibrotic activity on human embryonic fibroblasts / 中国中药杂志

In this study, 10 samples of parasites, cursive, and the whole from six different species of Cordyceps were determined and compared by HPLC and LC-MS methods. Uridine, adenosine, and cordycepin were selected as the main evaluation index. The anti-fibrotic activity of different species Cordyceps extracts was observed using in vitro TGF-β1-induced ECM accumulation in human embryonic fibroblasts CCC-ESF-1. The results demonstrated that the number of atoms and hyphae ingredients of different species showed little difference, however, the content distribution of each component has obvious significance. The in vitro anti-fibrotic activities of different species were as follow： Cordyceps flower > Cicada Cordyceps (Cicada flower)> Silkworm Cordyceps> Tussah Cordyceps>natural Cordyceps. Our preliminary data could serve as reference for the discovery of artificial alternatives of natural Cordyceps.

Pharmacokinetics of 8-O-acetylharpagide and harpagide after oral administration of Ajuga decumbens extract in beagle dog / 中国中药杂志

8-O-acetylharpagide and harpagide are two kinds of effective component of Ajuga decumbens extract. A sensitive LC-MS/MS method has been established for pharmacokinetics of 8-O-acetylharpagide and harpagide in beagle dog after oral administration of from A. decumbens extract. Female beagle dogs received orally 12.9, 25.7 mg x kg(-1) p. o. Concentrations of 8-O-acetylharpagide and harpagide in plasma were determined by LC-MS/MS method at different time points and all pharmacokinetic parameters were estimated by non-compartment analysis. The mobile phase consisted of 0.1% formic acid in water (A) and acetonitrile (B), which was run at a flow rate of 0.3 mL x min(-1). Chromatographic separation was achieved on an Agilent ZORBAX XDB-C18 column (2.1 mm x 50 mm, 3.5 microm) using a gradient elution of 5% B at 0-2 min, 95% B at 2. 1-5 min and 5% B at 5. 1-10 min. All analytes, including the IS, were monitored under positive ionization conditions and quantified in MRM mode with transitions of m/z 429.2-369.2 for 8-O-acetylharpagide, m/z 387.2-207.2 for harpagide, and m/z 149.2-103.1 for IS. High purity nitrogen was employed as both the nebulizing and drying gas. Other parameters of the mass spectrometer were optimized as follows: drying gas flow 10.0 L x min(-1); drying gas temperature 300 degrees C; capillary voltage 4 000 V. Results showed that 8-O-acetylharpagide and harpagide showed a dose-dependence profile. T(max) of 8-O-acetylharpagide is 1.7 h, and T(max) of harpagide is 1.57 h, which was higher than T(max) of 8-O-acetylharpagide and harpagide after oral administration of from A. decumbens extract in rats. The different pharmacokinetic parameters may be due to the species differences of rat and beagle dog.

Clinical and cardiac imaging characteristics of patients with left ventricular apical hypoplasia / 中华心血管病杂志

<p><b>OBJECTIVE</b>To analyze the clinical and cardiac imaging characteristics of patients with left ventricular apical hypoplasia (LVAH).</p><p><b>METHODS</b>From January 2008 to January 2012, seven patients [3 male/4 female, age: 6 - 44 (19.9 ± 14.2) years] with LVAH were included in this cohort. Transthoracic echocardiogram was performed in all patients, cardiovascular MRI was performed in 3 patients and cardiovascular CT in another 2 patients. In addition, one LVAH patient underwent cardiac catheterization and angiography examination.</p><p><b>RESULTS</b>Four out of 7 patients complained chest discomfort. Precordial murmur was heard in 3 patients. Atrial fibrillation was evidenced by electrocardiogram in 3 patients. Left ventricular end-diastolic diameter [(57.9 ± 11.6) mm] increased while left ventricule (LV) longitudinal diameter reduced in all patients. Left ventricular systolic function was reduced in 2 patients and mean LVEF was (47.6 ± 17.2)%. The interventricular septum bulged towards the right, and the ventricular septum thickness was (7.3 ± 1.2) mm. The papillary muscles were dominant on the flattened LV anteroapical region. The right ventricle elongated and wrapped around the hypoplastic left ventricular apex, and the dimension of right ventricle was (19.7 ± 7.6) mm. Focal fat replacement of the left ventricular apical wall was evidenced in 5 patients underwent cardiovascular MRI or CT examinations.</p><p><b>CONCLUSIONS</b>Clinical symptoms are non-specific in LVAH patients. Truncated and spherical LV, abnormal origin of papillary muscles in the flattened LV anterior apex and an elongated right ventricle wrapping around the LV apex as well as focal fat replacement of the left ventricular apical wall are typical imaging characteristics of LVAH.</p>

Fabrication and evaluation of the enzyme immunosensor for rapid detection of Vibrio parahaemolyticus based on chitosan-SiO2 hybrid membrane / 中华预防医学杂志

<p><b>OBJECTIVE</b>To improve the key technology of immunesensors in immobilizing bio-sensitive element and keeping its bioactivity, an enzyme immunosensor based on chitosan-SiO(2) (CS-Sio(2)) hybrid membrane was fabricated. To estimate the new immunosensor Vibrio parahaemolyticus which was the main pathogens of aquatic products.</p><p><b>METHODS</b>A CS-SiO(2) hybrid membrane was prepared using sol-gel method. The enzyme immunosensor was fabricated by coating the membrane and horseradish peroxidase labeled Vibrio parahaemolyticus antibody (HRP-anti-VP) on the surface of four-channel screen-printed carbon electrode. The immunosensor was characterized by cyclic voltammetry. Vibrio parahaemolyticus could be detected according to the decrease percentage (DP) of peak current before and after immune response, while cyclic voltammetry was used as an electrochemical mean to detect the products of the enzymatic reaction. Seven kinds of bacteria, like Vibrio alginolyticus, were selected for specific experiments.</p><p><b>RESULTS</b>By studying the infrared spectrum of three kinds of films, the CS-SiO(2) hybrid membrane was prepared and HRP-anti-VP was fixed in the hybrid membrane. Under the optimum conditions of immunoreaction and electrochemical detection, the DP of peak current before and after immune response showed a linear relation with lgC in the range of 10(4) - 10(9) cfu/ml, while the linear regression equation was: DP = 6.5 lgC-3.319, the correlation coefficient was 0.9958 and the detection limit was 6.9 x 10(3) cfu/ml (S/N = 3). The immunosensor possessed acceptable specificity, reproducibility (RSD < 6%), stability (the amperometric response was 95% of the initial response after a week) and accuracy (96.7% of the results obtained by the immunosensor were in agreement with those obtained by GB/T 4789.7-2003).</p><p><b>CONCLUSION</b>The enzyme immunosensor based on CS-SiO(2) hybrid membrane gave a good performance in rapid detection of Vibrio parahaemolyticus.</p>

Studies on protective effects of Niuhuang Shangqing pills on cerebral ischemia / 中国中药杂志

<p><b>OBJECTIVE</b>To discuss the protective effect of Niuhuang Shangqing pills (NSP) on the experimental cerebral ischemia and the influence of blood rheology in animals.</p><p><b>METHOD</b>Using the middle cerebral artery occlusion (MCAO), bilateral common carotid artery ligation and lipopolysaccharide (LPS) in combination with carrogeenin (Ca)-induced stagnation of blood model rats, we investigated the influence of NSP on the physical sign indexes, brain infarct size and the water content, the content of lactate(LD), glutathione peroxidase (GPx), catalase (CAT), malondialdehyde (MDA) and the blood rheology.</p><p><b>RESULT</b>The 3.00, 1.50 g x kg(-1) dosage groups of NSP decreased the neurosigns indexes, significantly reduced the brain infarct areas in MCAO rats, decreased the brain water content, increased the activities of GPx and CAT, decreased the content of LD and MDA in bilateral common carotid artery ligation rats. They improved the blood rheology in lipopolysaccharide (LPS) in combination with carrogeenin (Ca)-induced stagnation of blood model rats.</p><p><b>CONCLUSION</b>NSP has the function of protective effect on experimental experimental cerebral ischemia.</p>

Improvement effects of Jincao tablet on immune function of model of hysteromyoma in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the improvement effects of Jincao tablet on immune function of the model of hysteromyoma in rat and the relationship between the model and pathogenesis.</p><p><b>METHOD</b>Rats were randomly divided into 6 groups: normal group, model group, treatment groups including low,middle and high dosage groups of Jincao tablet and Guizhi Fuling pill. Rats were injected respectively with diethyl stilbestrol and progesterone. The immune apparatus of rats were measured. The levels of CD3, CD4 and CD4/ CD8 in serum were determined by flow cytometer. The estrogen and receptor were measured by radioligand binding assay and pathologic changes of womb tissue were observed microscopically.</p><p><b>RESULT</b>Compared with normal group, the weight of thymus, the levels of CD3, CD4 and CD4/CD8 of model group were significantly decreased, and the levels of estrogen, estrogen receptor and CD8 were obviously increased. Jincao tablet groups were significant difference compared with model group and could alleviate the pathological changes of womb tissue.</p><p><b>CONCLUSION</b>Jincao tablet could improve the levels of immune function of the model of hysteromyoma in rat, and it might play a role in the pathogenesis of leiomyoma.</p>

Determination of acetylharpagide in Ajuga decumbens by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a method of the quantitative determination of acetylharpagide in Ajuga decumbens.</p><p><b>METHOD</b>The chromatographic conditions were as follows: a Phenomenex Luna C18 column was used, the mobile phase was composed of acetontrile-water (15:85), the flow rate was 1.0 mL x min(-1) and the UV absorbance detection was set at 197 nm.</p><p><b>RESULT</b>Linearity of acetylharpagide was in the range of 0.6-3.6 microg (r = 0.9993), and the average recovery and RSD were 99.13% and 2.48%, respectively.</p><p><b>CONCLUSION</b>The contents of acetylharpagide ranged 0.40%-6.39% in A. decumbens. The method was simple, accurate and sensitive.</p>

Study on the metabolism of the precipitation of Xiexin decoction in rats I / 中国中药杂志

<p><b>OBJECTIVE</b>To study the process in vivo of the components in the precipitation of Xiexin decoction.</p><p><b>METHOD</b>The components in the rats' urine samples after oral administration of baicalin, single herb decoction of Radix Scutellariae or the precipitation of Xiexin decoction were analyzed by LC-MS-MS method; and the absorption of baicalin in different samples were calculated by intestinal in situ loop method.</p><p><b>RESULT</b>The urinary excretion amounts of baicalin in three samples were not obviously different, whereas the time reach elimination-peak of baicalin in them had significant difference. The absorption of baicalin in the precipitation was obviously greater than that in the single herb decoction and single baicalin. We found that wogonoside and 7-O-glucuronide chrysin were the metabolites presenting in the rat urine samples after oral administration of baicalin by LC-MS-MS, which had never been reported.</p><p><b>CONCLUSION</b>The resident time in the body of baicalin in the precipitation is prolonged, compared with that in the single herb decoction and single baicalin. The metabolites may be the potential biological components in vivo of baicalin in the precipitation.</p>

Expression of the infectious bursal disease virus polyprotein in Vero cells using attenuated Salmonella typhimurium as transgenic carrier / 生物工程学报

To examine if polyprotein gene (VP2/VP4/VP3) of Infectious Bursal Disease Virus (IBDV) could be delivered into mammalian cells and expressed using attenuated Salmonella typhimurium as vector. The IBDV polyprotein gene was amplified by RT-PCR and inserted in to pCI, an eukaryotic expression plasmid. The resulting recombinant pCI-VP2/VP4/VP3 was transformed by electroporation into attenuated Salmonella typhimurium strain ZJ111 (dam- and phoP-), which was then use to transfect the Vero cells. Gene specific RT-PCR revealed that VP2/VP4/VP3 was transcribed into mRNA in the Vero cells. Indirect immunofluorscence assay, SDS-PAGE and Western-blot analysis showed that VP2/VP4/VP3 was expressed and the product was immuno-reactive with anti-IBDV serum. This work provides essential precondition for developing a new oral DNA vaccine against IBDV.

Expression of polyprotein of infectious bursal disease virus in Bombyx mori / 生物工程学报

Segment A of the genome of infectious bursal disease virus(IBDV) encodes structure protein VP2 and VP3 and protease VP4. In this study a polyprotein gene of IBDV was inserted into a Bombyx mori baculovirus transfer vector pAcHLT--C and contransfected into BmN cells with linear genome DNA of virus Bm-BacPAK6. Dot hybridization suggested that the segment A of the virus genome was inserted in the genome of Bm-BacPAK6. The silkworm of fifth instars were infected by the recombinant virus and the immunogenicity of the infected larvae's blood were examined with ELISA, SDS-PAGE and Western blotting. It appears that the recombinant polyprotein has the property of immunoreactivity and the expression in larvae reached the pick 5-day post infection.