Characterization of potential bitterness critical quality attributes in Xiaoer Ganmao Oral Liquid based on UHPLC-LTQ-Orbitrap MSn / 药学学报

To identify the bitter compounds of real-world Xiaoer Ganmao Oral Liquid sugar-free intermediates, an integrated strategy has been developed by using ultra-high performance liquid chromatography with linear ion trap-Orbitrap mass spectrometry (UHPLC-LTQ-Orbitrap MSn) method and BitterX database prediction. The chromatographic operating conditions were as follows, chromatographic column: Acquity UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm), mobile phase: 0.1% formic acid-water solution (A)-acetonitrile (B) with gradient elution. The data were collected in positive and negative ion modes, respectively. The accurate molecular mass and structural information of the target compounds were obtained based on quasi-molecular ions and fragmentation ions provided by high-resolution mass spectrometry. The compounds were identified by combining retention time, reference substances, reports, and other relevant data, and a total of 57 constituents including flavonoids, alkaloids, and phenylpropanoids were finally identified. Further, the BitterX database was used to predict binding probability of compounds to bitter receptors and identify potential bitter critical quality attributes, finally 33 potential bitter compounds, including kukoamine A and linarin, were predicted. This study comprehensively characterized the material basis of Xiaoer Ganmao Oral Liquid sugar-free intermediates, it provides an effective method for bitter compound screening and a reference for further improving the undesirable taste of Xiaoer Ganmao Oral Liquid.

Blood matching incompatibility caused by cold autoantibodies and IgG anti S antibody: a case report / 中国输血杂志

【Objective】 To analyze the serological characteristics of an anemia patient with cold autoantibodies and IgG anti S antibody. 【Methods】 Unexpected antibody screening was performed on screening cells, autologous cells and panel cells with patient serum in saline and anti human globulin card media at room temperature and 37 ℃, respectively. At the same time, the serum that destroying IgM antibodies and acid elution solution were respectively screened for unexpected antibodies in anti human globulin card media. The absorption-elution test was used to treat patients′ red blood cells and serum, and the unexpected antibody identification and cross matching were performed after excluding the interference of cold autoantibodies. 【Results】 Cold autoantibodies and IgG anti S antibody were found in the serum of this patient. 【Conclusion】 When a patient is found to have cold autoantibodies, it is necessary to screen the alloantibody, in order to avoid post transfusion immune reactions due to missed detection of alloantibody.

Investigation on therapeutic potential and molecular mechanisms of Kunxian capsule against diabetic kidney disease via reversing the imbalance of "immune-inflammation" network / 药学学报

Drug repositioning provides new clinical indications for existing drugs. The imbalance between body's "immune-inflammation" regulation is one of the important factors in the occurrence and development of diabetic nephropathy (DN). Chinese patent medicine Kunxian capsule is clinically used for treating rheumatoid arthritis with satisfying immune-modulatory and anti-inflammatory actions. Notably, accumulating clinical evidence based on small cohorts had shown that Kunxian capsule may be used to treat DN. But the underlying pharmacological mechanisms remain unclear. Therefore, this study integrated "drug target-disease gene-biological pathway-function module" multi-level associated network analysis, and in vivo and in vitro experiments, to verify the pharmacological effects of Kunxian capsules in DN and to elucidate its molecular mechanisms. The experimental protocol was reviewed by the Laboratory Animal Welfare and Ethics Committee of China Academy of Chinese Medical Sciences, and it complies with the relevant regulations on laboratory animal welfare and ethics. As a result, the network analysis showed that the candidate targets of Kunxian capsule against DN were significantly involved into various functional modules which were related to modulation of immune-inflammation system, basement membrane lesion, abnormal hemorheology, energy metabolism and hormone metabolism, and the number of targets enriched by PI3K/AKT/NF-κB pathway is the largest. In addition, both in vivo and in vitro experiments demonstrated that Kunxian capsule by gavage effectively reduced blood glucose, improved insulin resistance, reduced blood lipid, inhibited renal extracellular matrix protein production and renal inflammation, improved renal function and pathological damages, and inhibited the activity of PI3K/AKT/NF-κB/TNF-α/IL-1β pathway in diabetic nephropathy rats. Collectively, these findings suggest the therapeutic potentials of Kunxian capsule to alleviate DN by regulating the imbalance of immune-inflammation system.

Proteomic characterization of four subtypes of M2 macrophages derived from human THP-1 cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Macrophages are widely distributed immune cells that contribute to tissue homeostasis. Human THP-1 cells have been widely used in various macrophage-associated studies, especially those involving pro-inflammatory M1 and anti-inflammatory M2 phenotypes. However, the molecular characterization of four M2 subtypes (M2a, M2b, M2c, and M2d) derived from THP-1 has not been fully investigated. In this study, we systematically analyzed the protein expression profiles of human THP-1-derived macrophages (M0, M1, M2a, M2b, M2c, and M2d) using quantitative proteomics approaches. The commonly and specially regulated proteins of the four M2 subtypes and their potential biological functions were further investigated. The results showed that M2a and M2b, and M2c and M2d have very similar protein expression profiles. These data could serve as an important resource for studies of macrophages using THP-1 cells, and provide a reference to distinguish different M2 subtypes in macrophage-associated diseases for subsequent clinical research.

The health benefits of the active ingredients and whole food components of natural plants through regulating the flora / 生物工程学报

With the cooperation of bacteria and the human body, the nutrients in food are deeply digested, utilized, and shared. In addition, symbiosis is formed between microorganisms and hosts. Such a delicate combination makes the microorganisms form the inherent flora in the human body. They obtain the biological basis for survival, and provide the necessary regulation and support for the host in terms of immunity and nutrition, through their functional metabolism and population signals. At present, most of the researches focus on the isolation and evaluation of the functional components of plants, such as plant polysaccharides, polyphenols, flavonoids, and other active functional components. However, in traditional Chinese medicine, plants are often used with whole food components. To date, studies have found that the dynamics of flora affecting human health are not fixed, nor dependent on the change of a single strain. The ecological competition and metabolic regulation between microorganisms are usually coevolved with the host. The regulatory effect of natural plants for both medicine and food mainly depends on their whole food components. This provides evidence to support the role of whole food components played in promoting the efficacy of traditional Chinese medicine from the perspective of microenvironment. Therefore, the development and utilization of medicinal and edible natural plant activities should be fully understood and evaluated with flora regulation.

Parotid CT imaging reporting and data system: A preliminary study / 北京大学学报(医学版)

OBJECTIVE@#To establish a Parotid Imaging Reporting and Data System (PI-RADS) for CT diagnosis of the parotid gland neoplasms and to investigate the clinical applicable value and feasibility of PI-RADS.@*METHODS@#Patients who had been diagnosed with primary parotid gland neoplasms and had received surgical treatments in Peking University School and Hospital of Stomatology during the period of January 2013 to December 2016 were included in this study. The diagnoses were confirmed by the postoperative pathological examinations in all the patients. The CT imaging data of all patients were retrospectively reviewed and analyzed by two readers in consensus. Imaging characteristics related to the parotid neoplasms were extracted and quantified. Based on comprehensive analysis of the imaging characteristics, the probabilities of the benign and malignant neoplasms were evaluated and classified into six grades, PI-RADS 1-6 (PI-RADS 1: normal parotid gland; PI-RADS 2: confidently benign lesions; PI-RADS 3: probably benign lesions without confirmed evidence of malignancy; PI-RADS 4: suspected malignancy without sufficient evidence of malignancy; PI-RADS 5: confidently malignant lesions; PI-RADS 6: lesions with confirmed pathological evidence of malignancy).@*RESULTS@#A total of 897 patients with 1 003 parotid lesions were included. The lesions included 905 benign and 98 malignant lesions. The proportions of the malignancies in PI-RADS 2, PI-RADS 3, PI-RADS 4 and PI-RADS 5 according to the two readers in consensus were 0.4%, 5.7%, 35.5% and 96.7% respectively. The overall Cohen's Kappa test showed medium consistency between the two independent researchers (κ=0.614, P<0.001, 95%CI: 0.569-0.695). Pearson Chi-square test showed that the proportions of malignancies increased with the diagnostic PI-RADS grades (Cochran-Armitage trend test, Z=-15.579, P<0.001). The results of Pearson Chi-square tests showed significant differences between the grades [PI-RADS 2 and 3 (χ²=12.048, P=0.001); PI-RADS 3 and 4 (χ²=75.231, P<0.001); PI-RADS 4 and 5 (χ²=32.266, P<0.001)].@*CONCLUSION@#PI-RADS can be used to evaluate the risk of malignancy and will be helpful to improve the imaging diagnosis and clinical treatment of parotid gland neoplasms.

Quantification of aristolochic acids and their DNA adducts in mice kidney and liver by HPLC-MS/MS / 中国中药杂志

This study aims to establish a quantitative method of 4 aristolochic acids-DNA adducts in mice kidney and liver based on high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) for monitoring the content changes of aristolochic acids-DNA adducts. A Shiseido Capcellpak AQ C_(18) column(3 mm×100 mm, 3 μm) was used, with a mixture of 0.2% acetic acid-5 mmol·L~(-1) ammonium acetate as the aqueous phase and methanol as the organic phase for gradient elution. The multiple reaction monitoring(MRM) scanning method under positive mode by electrospray ionization(ESI) was performed for the detection of the aristolochic acids-DNA adducts which formed by combining aristolochic acid Ⅰ/Ⅱ with deoxyadenosine, deoxyguanosine, and deoxycytidine, respectively. Balb/c mice were given Guanmutong extract by gavage, and the relative content of aristolochic acids-DNA adducts in liver and kidney samples were analyzed within 60 days. It was found that the concentration of 4 aristolochic acids-DNA adducts in the kidney was significantly higher than that in the liver, and there were about 15.87 adducts in per 1×10~6 normal deoxynucleosides, which was 4.5-7.5 times than that of the liver. What's more, some adducts can still be detected on the 30 th day after administration. The concentration of the adducts in the liver was highest on the first day after administration, and a second peak appeared during the 7 th to 14 th days. The results indicated that aristolochic acids-DNA adducts are difficult to eliminate in vivo, and it is of great significance to study the mechanism of liver and kidney injury of aristolochic acid.

Variation analysis of genomic methylation induced by high temperature stress in Pinellia ternata / 中国中药杂志

Pinellia ternata belongs to the Araceae family and is a medicinal herb. The tuber is the medicinal organ with antitussive, antiemetic and anti-tumor activities. It is easy to encounter high temperature environment during the growth periods, leading to decrease of tuber production. At present, the mechanism of response to high temperature stress in P. ternata is still unknown. DNA methylation plays a vital role in plant protection against adversity stress as a way of epigenetic regulation. In this study, P. ternata was used as material for treatment of high temperature stress at 0 h, 6 h and 80 h, and methylation sensitive amplification polymorphism(MSAP) analysis was conducted on the changes of DNA methylation in its genome. The results showed that 20 pairs of MSAP primers were selected from 100 MSAP primers with multiple clear and uniform bands, and 353, 355 and 342 loci were amplified from materials of P. ternata treated in the high temperature stress 0 h, 6 h and 80 h, respectively. Cytosine methylation levels of CCGG context in the above materials were characterized as 60.91%, 44.79% and 44.74%, respectively. And the full methylation ratios were 16.71%, 22.25% and 29.24, respectively. It demonstrated that high temperature stress significantly induced the down-regulation of DNA methylation level and up-regulation of the full methylation rate in P. ternata genome. This study provides a preliminary theoretical reference for analyzing the mechanism of P. ternata responding to high temperature stress from the epigenetic perspective.

Computed tomographic features of desmoplastic ameloblastoma of the jaw / 北京大学学报(医学版)

OBJECTIVE@#To retrospectively analyze the clinical and CT imaging features of desmoplastic ameloblastomas (DA) in comparison with other types of ameloblastomas.@*METHODS@#Patients diagnosed with ameloblastoma histopathologically in Peking University School and Hospital of Stomatology from July 2000 to August 2017 were reviewed to analyze the constituent ratios and characteristics of DA. CT imaging features of DA (28 cases) were investigated in comparison with consecutive cases of solid/multicystic and unicystic ameloblastomas. The following imaging perspectives were analyzed: the border, internal structure, three-dimensional shape, tooth, the periodontium, the cortex and the expansion of the tumors. CT values were measured in the region of interest for quantitative analysis.@*RESULTS@#Among the 1 269 cases of ameloblastomas, 50 (3.9%) were DA, which showed predilections for males (74.0%). The average age of DA was about 43.9 years old, which was higher than the other two types. The anterior incisorcanine region (62.0%) and premolar region (30.0%) were most frequently affected. The incidence rate of DA in mandibule was 56.0% (28/50), which was slightly higher than that of maxilla (44.0%). The DA characteristically showed scallop border and honeycomblike or soapbubble internal structure with bone formation on CT. The mean ratios of height to mesiodistal and buccal-lingual to mesio-distal distances were 0.76 and 0.63, which were higher than the other two types. According to the degree of internal bone formation, three subtypes of DA could be observed: densely ossifying type (I), honeycomb/soap bubble type (II) and sparsely ossifying type (III). The means and standard deviations of CT values of DA were significantly higher than those of the other two types, which were (488.8±164.0) HU (type I), (171.7±102.8) HU (type II), (42.1±8.8) HU (type III).@*CONCLUSION@#CT is helpful for diagnosis of DA, which shows as solid tumor with varying degrees of internal ossification.

Determination of 25 characteristic components in Danhe Granules by LC-MS and consistency analysis of preparations / 中草药

Objective: A high performance liquid chromatography coupled to triple quadrupole mass spectrometry method (HPLC- MS/MS) was established to simultaneously determine the content of 25 characteristic components (gallic acid, tanshinol, catechin, chlorogenic acid, caffeic acid, epicatechin, rutin, polydatin, hyperin, astragalin, naringin, hesperidin, rosmarinic acid, resveratrol, salvianolic acid B, quercetin, emodin-8-O-β-D-glucoside, isorhamnetin, emodin, coclaurine, nuciferine, cryptotanshinone, tanshinone I, dehydronuciferine, tanshinone IIA) in Danhe Granules (DG), and the consistency between different batches was investigated. Methods The analysis was conducted on Agilent Rapid Resolution HD C18 column (50 mm × 2.1 mm, 1.8 μm) with the mobile phase of 0.1% formic acid water-0.1% formic acid acetonitrile. The dynamic multi-response detection (dMRM) scanning mode was used in the mass spectrometry. Results: Based on the established HPLC-MS/MS method, the simultaneous quantitative analysis of 25 characteristic components could be completed within 10 min, with the quantitative limits of isorhamnetin, coclaurine, and dehydronuciferine of 0.025 ng/mL; emodin and tanshinone I of 0.050 ng/mL; emodin-8-O-β-D-glucoside of 0.200 ng/ml; gallic acid, chlorogenic acid, caffeic acid, epicatechin, rutin, hyperin, astragalin, resveratrol, quercetin, nuciferine, tanshinone IIA of 0.250 ng/ml; catechin, rosmarinic acid, and cryptotanshinone of 0.500 ng/mL; tanshinol, hesperidin, and salvianolic acid B of 1.000 ng/mL; polydatin of 2.000 ng/mL; naringin of 5.000 ng/mL, respectively. The linear relationships of the 25 constituents within their respective mass concentrations were good, with the average recovery of 85.16%-113.46% and the RSD of 2.01%-8.80%. Furthermore, this method also included the main components named monarch, minister, assistant and guide of herbs in a relatively comprehensive way. The total content of the 25 components was 31.49 mg/g, among which the content of salvianolic acid B (9.44 mg/g) and hesperidin (7.60 mg/g) was the highest, and the content of isorhamnetin (0.79 μg/g) was the lowest. According to boxplot analysis, the content of 25 components in 10 different batches of DG fluctuated (P value) within 75% < P < 125%; and the RSD value of 25 components ranged from 2.58% to 13.10% by statistical analysis. The above results showed that the consistency of the component content among 10 batches of DG was acceptable. Conclusion: The analytical method established in this study is fast and sensitive. Furthermore, the results of this study are reliable and can provide scientific methods and basis for quality control and consistency analysis of DG.

Positive Effects of Qing'e Pill () on Trabecular Microarchitecture and its Mechanical Properties in Osteopenic Ovariectomised Mice / 中国结合医学杂志

OBJECTIVE@#To investigate the impact of Qing'e Pill (, QEP) on the cancellous bone microstructure and its effect on the level of β-catenin in a mouse model of postmenopausal osteoporosis.@*METHODS@#Ninety-six 8-week-old specific pathogen free C57BL/6 mice were randomly divided into 4 groups (24/group): sham, ovariectomised osteoporosis model, oestradiol-treated, and QEP-treated groups. Three months after surgery, the third lumbar vertebra and left femur of the animals were dissected and scanned using micro-computed tomography (micro-CT) to acquire three-dimensional (3D) parameters of their cancellous bone microstructure. The impact of ovariectomy, the effect of oestradiol and QEP intervention on cancellous bone microstructure, and the expression of β-catenin were evaluated.@*RESULTS@#The oestradioland the QEP-treated groups exhibited a significant increase in the bone volume fraction, trabecular number, trabecular thickneßs, bone surface to bone volume ratio (BS/BV), and β-catenin expression compared with those of the model group (P <0.05). In contrast, the structure model index, trabecular separation, and BS/BV were significantly decreased compared with those of the ovariectomised osteoporosis model group (P <0.05). No differences were observed in the above parameters between animals of the QEP- and oestradiol-treated groups.@*CONCLUSIONS@#The increased β-catenin expression may be the mechanism underlying QEP's improvement of the cancellous bone microstructure in ovariectomised mice. Our findings provide a scientific rationale for using QEP as a dietary supplement to prevent bone loss in postmenopausal women.

Association between ADAMTS14 gene polymorphism and the temporomandibular joint osteoarthritis in Chinese Han females / 北京大学学报(医学版)

OBJECTIVE@#To investigate the association between single nucleotide polymorphisms (SNP) of ADAMTS14 gene rs4747096 and osteoarthritis of the temporomandibular joint in Chinese Han females.@*METHODS@#As a case-control study, a total of 213 Chinese Han females were involved in the present study, which contained 103 temporomandibular joint osteoarthritis patients and 110 healthy people who had no symptoms or signs of temporomandibular joint osteoarthritis as control. Peripheral blood samples were collected from each participant. Genomic DNAs of temporomandibular joint osteoarthritis patients and healthy control were extracted from peripheral venous blood, which were stored in -80 °C refrigerator by using DNA extraction kits. The designed primers were used for polymerase chain reaction (PCR) amplification of specific DNA fragments. Genotype was determined by sequencing the PCR products. The software Chromas 2.22 was used to analyze the genotype. The genotype distributions, allele frequencies and genetic models between the patients and controls were compared. The age distribution was checked by t-test. Genotype and allele frequency were detected by Chi-square test.@*RESULTS@#In the present study, there were no significant differences between the osteoarthritis patients and healthy controls in terms of age. The genotype distribution was in accordance with Hardy-Weinberg equilibrium in the two groups. The genotype frequency of the ADAMTS14 (rs4747096) in the experimental group was 38.8% (AA), 55.4% (AG), and 5.8% (GG), respectively. The genotype frequency in the control group was 40.9% (AA), 43.6% (AG), and 15.5% (GG), respectively. The difference of genotype frequency of the ADAMTS14 (rs4747096) was significant between the experimental group and the control group (P=0.047). There was no significant difference in allele frequency between the two groups (P=0.415). AA and AG genotypes significantly increased the risk of the disease compared with GG in dominant model (OR=1.114, 95% CI: 1.015-1.223, P=0.028).@*CONCLUSION@#A significant correlationship was found between the ADAMTS14 (rs4747096) SNP and the temporomandibular joint osteoarthritis in Chinese Han females. The distribution of rs4747096 may be different between temporomandibular joint osteoarthritis and healthy population.

Protective effect and mechanism of Lactobacillus on cerebral ischemia reperfusion injury in rats

The present study was designed to investigate the protective effects and mechanism of inactivated lactobacillus (ILA) on cerebral ischemia reperfusion injury (CIRI) in rats. In this experiment, 30 male Sprague Dawley rats were randomly divided into control group, IRI groups, and ILA group. A middle cerebral artery occlusion and reperfusion model was prepared. The rats were killed after 24 hours of recovery of blood flow of cerebral ischemia resulting from 60-min occlusion. The cerebral infarction volume and neurological scores were assayed by staining and behavioral observation. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels were assayed by biochemical kits. Cell apoptosis was assayed by Tunnel and the Toll-like receptor (TLR)-4, IkB, and A20 were assayed by western blot. The neurobehavioral scores in IRI rats were significantly lower compared to the control group while ILA improved the neurobehavioral scores of the ILA groups. The cerebral infarction volume and neural cell apoptosis of rats in the ILA groups decreased significantly compared with those in the IRI group. In addition, MDA level in the ILA groups decreased whereas SOD activity increased compared to the IRI group. Moreover, ILA also inhibited the expression of TLR-4 and promoted the expression of IkB and A20. ILA inhibited the apoptosis of neural cells, decreased cerebral infarction volume, and reduced oxidative stress through inhibition of TLR-4/NF-kappa B signaling, improving neurobehavioral scores. Thus from the present study it was concluded that ILA has protective effect on CIRI.

Molecular and Epidemiological Characterization of Infant Botulism in Beijing, China / 生物医学与环境科学（英文）

Laboratory-based pathogen isolation, identification, and toxicity determination were performed on samples from a suspected case of infant botulism. Mice injected with cultures generated from the enema sample and ingested Powered infant formula (PIF) presented typical signs of botulism. Antitoxins to polyvalent botulinum neurotoxins (BoNTs) and monovalent BoNT type B antitoxin had protective effects. Clostridium botulinum isolated from the enema and residual PIF samples were positive for type B toxin. Pulsed-field gel electrophoresis (PFGE) revealed that the two strains of C. botulinum isolated from the two samples produced indistinguishable pulsotypes. These findings confirmed this case of type B infant botulism associated with the ingestion of PIF contaminated by type B C. botulinum spores.

Effects of Modified Qing'e Pill () on expression of adiponectin, bone morphogenetic protein 2 and coagulation-related factors in patients with nontraumatic osteonecrosis of femoral head / 中国结合医学杂志

<p><b>OBJECTIVES</b>To observe the regulation of Chinese herbal medicine, Modifified Qing'e Pill (, MQEP), on the expression of adiponectin, bone morphogenetic protein 2 (BMP2), osteoprotegerin (OPG) and other potentially relevant risk factors in patients with nontraumatic osteonecrosis of the femoral head (ONFH).</p><p><b>METHODS</b>A total of 96 patients with nontraumatic ONFH were unequal randomly divided into treatment group (60 cases) and control group (36 cases). The treatment group were treated with MQEP while the control group were treated with simulated pills. Both groups were given caltrate D. Six months were taken as a treatment course. Patients were followed up every 2 months. The levels of plasma adiponectin, BMP2, OPG, von Willebrand factor (vWF), von Willebrand factor cleaving protease (vWF-cp), plasminogen activator inhibitor 1 (PAI-1), tissue plasminogen activator (tPA), C-reactive protein (CRP), blood rheology, bone mineral density (BMD) of the femoral head and Harris Hip Score were measured before and after treatment.</p><p><b>RESULTS</b>After 6 months of treatment, compared with the control group, patients in the treatment group had signifificantly higher adiponectin and BMP2 levels (P<0.01 and P=0.013, respectively), lower vWF, PAI-1 and CRP levels (P=0.019, P<0.01 and P<0.01, respectively), and lower blood rheology parameters. BMD of the femoral neck, triangle area and Harris Hip Score in the treatment group were signifificantly higher than those in the control group. Moreover, plasma adiponectin showed a positive association with BMP2 (r=0.231, P=0.003) and a negative association with PAI-1 (r=-0.159, P<0.05).</p><p><b>CONCLUSION</b>MQEP may play a protective role against nontraumatic ONFH by increasing the expression of adiponectin, regulating bone metabolism and improving the hypercoagulation state, which may provide an experimental base for its clinical effects.</p>

Effect of TPh on cell HepG-2 cell cycle, apoptosis and related gene expression of hepatocellular carcinoma / 中国生化药物杂志

Objective To investigate the effect of different concentrations of TPh on hepatoma cells , cell viability and and its possible mechanisms of anti-tumor.Methods The inhibitory rate of hepatoma cells and cell viability on different concentrations of TPh and time were measured by MTT assay;The morphological changes of apoptosis were observed by Hoechst 33258 straining; Cell cycle distribution was evaluated by flow cytometry (FCM).The protein expression of Bcl-2 and Bax were detected by Western blot analysis.Results MTT assay showed that TPh inhibited the proliferation of HepG-2 cells in a dose-dependent manner, and the inhibitory rate increased with the increase of concentration.The inhibitory rate was 50.9% (P<0.01).The results of Hoechst 33258 staining showed that the fluorescence intensity of hepatocellular carcinoma cells was light blue in fluorescence microscopy and bright blue fluorescence in apoptotic hepatocarcinoma cells, and the apoptosis of hepatocarcinoma cells increased with the increase of drug concentration.The percentage of cells in G0/G1 phase increased with the increase of cell cycle, and the ratio of cells in S phase was decreased in G2/M phase compared with blank control group (P<0.05);Western blot results showed that compared with the blank control group, TPh inhibited the proliferation of Bcl-2 cells in a concentration-dependent manner ( P <0.05 ) , and the number of apoptotic cells increased with the increase of drug concentration (P<0.05), and the ratio of Bcl-2/Bax in the TPh group decreased significantly (P<0.05), and the expression of Bax gene increased. Conclusion TPh inhibits cell proliferation, promotes apoptosis and induces HepG-2 to block G0/G1 phase.Its mechanism may increase the expression of Bax and decrease Bcl-2 protein expression.

Effects of B7-H3 on T lymphocytes in different activation states / 中华微生物学和免疫学杂志

Objective To study the impacts of B7-H3 molecule on the proliferation of T lymphocytes in different activation conditions and on the secretion of relevant cytokines.Methods Peripheral blood samples were collected from healthy subjects to separate peripheral blood mononuclear cells (PBMC) by Ficoll density gradient centrifugation.T lymphocytes were isolated from some of the PBMCs and purified with T Cell Enrichment Kit.PBMC and purified T lymphocytes were activated by anti-CD3/CD28 monoclonal antibodies (McAb) in vitro.Flow cytometry analysis was used to detect the expression of CD28 and CTLA-4 on T lymphocytes at different time points for further analyzing the activation states of T lymphocytes.On this basis, human B7-H3-Fc fusion protein was added into the mixed co-cultivation system on days 0, 1, 2, 3 and 4, and Hu-Fc fusion protein was used as isotype control.CCK-8 method was performed to detect the proliferation of T lymphocytes in each group.ELISA method was used to detect the secretion of cytokines (IL-2, IL-10 and IFN-γ) and to analyze the immune responses induced by stimulating T lymphocytes at different states of activation with B7-H3.Results B7-H3 molecule significantly inhibited the quiescent T lymphocytes from secreting IL-2 and IL-10, but had no significant impact on IFN-γ secretion.Moreover, it significantly promoted the activated T lymphocytes to secret IL-2 and IFN-γ, but had no obvious impact on IL-10 secretion.Results of the cell proliferation assay showed that B7-H3 molecule inhibited the in vitro proliferation of T lymphocytes in the PBMC, but had no obvious impact on purified T lymphocytes.ConclusionThe regulatory effects of B7-H3 molecule on the immune functions of T lymphocytes vary with the activation states of T lymphocytes.

Chemical constituents of Euodia rutaecarpa var. officinalis and their antifungal activities / 中草药

Objective: To study the chemical constituents from the fruits of Evodia rutaecarpa var. officinalis. Methods: The constituents were isolated and purified by chromatographic methods. The structures were elucidated by physicochemical properties and spectroscopic methods. The antifungal activities were tested with mycelium growth method. Results: Fifteen compounds were obtained from the 70% ethanol extract of E. rutaecarpa var. officinalis, and were identified as evodiamine (1), rutaecarpine (2), dehydroevodiamine (3), dihydroevocarpine (4), evocarpine (5), 1-methyl-2-undecyl-4 (1H)-quinolone (6), 1-methyl-2-[(4Z,7Z)- 4,7-tridecadienyl]-4 (1H)-quinolone (7), 1-methyl-2-[(6Z,9Z)-6,9-pentadecadienyl-4 (1H)-quinolone (8), limonin (9), shihulimonin A (10), isolimonexic acid (11), wuzhuyurutine B (12), stigmasterol (13), β-sitosterol (14), and β-daucosterin. (15). Conclusion: Compounds 5-8 and 11-13 are isolated from E. rutaecarpa var. officinalis for the first time. Different types of compounds show diverse antimicrobial activities against plant-pathogenic fungi.

The impact of separating drug sales from medical services reform on mechanism of controlling outpatient and emergency expenses in Beijing / 中国卫生政策研究

Objective:To evaluate the impact and analyze the mechanism of separating drug sales from medical services reform on the total medical expenditure containment of outpatient services in Beijing. Methods:The monitoring data and historical data from 89 third-level,78 second-level public hospitals,and 206 primary health institutions and community health service agencies,which participated in the reform,were analysed to estimate the impacts on outpa-tient service utilization and service price. The analysis was based on the data of the first and second quarters of 2016 outpatient and emergency department fee changes,and analysis of service utilization and service price factors on the im-pact of changes in outpatient and emergency services mechanism. Results:The total outpatients'expenditure in Beijing in the second quarter of 2017 was 17.38 billion yuan,representing a slight decrease of 1.14% than the expenditure in 2016. The growth rate reduced by 2.46%. Expenditure in third-level and second-level hospitals decreased by 2.82% and 3.02%,respectively,with reductions in growth rates of 4.06% and 3.74% respectively. Expenditure in primary institutions increased by 17.09%. The increase rate in the second quarter of 2017 was 26.28%,representing a remark-able increase compared to the rate in 2016(15.84%). The contribution from the medical institution service prices and service utilization to the change of outpatient and emergency medical expenditure containment was 6.98% and -7. 65%,respectively. For third-level hospitals,the contribution was 10.37% and -12.00%,respectively;second-level hospitals were 0.72% and -3.35%,respectively;and the primary health institutions was 6.08% and 9.57%. Con-clusions:Beijing's comprehensive reform of separating drug sales from medical services reform has effectively controlled the rising medical expenditure of outpatient services,mainly by adjusting outpatients service utilization between differ-ent level healthcare institutions.

Application of germ-free animals in the study of pathogenesis of cow mastitis / 中国实验动物学报

Mastitis is one of the high incidence diseases in dairy farm, and the traditional therapies include using of antibiotics which usually have a high recurrent rate. The disease recurrence will reduce the milk production and cause worse result such as breast necrosis. So many cows are obsoleted because of mastitis and that cause a great economic loss to the dairy industry. Nursing mothers also suffering from mastitis and they cannot be treated with antibiotics because the med-icine has a bad influence on the baby' s nervous system. With all of the negative effects, antibiotics cannot be used in the mastitis treatment. Therefore it is extremely urgent to develop a new therapy for mastitis. Probiotics is a viable alternative which can be used in the prevention and treatment of mastitis because of many successful applications. Many previous ex?amples in a variety of disease prevention and treatment using probiotics make this research very interesting. Digestive tract is the main niche where probiotics can play a crucial role in maintaining the host' s health. It has been observed that diges?tive tract is one of the important microbial niche, which have a robust relationship with mastitis. It is the major issue of this research to use probiotic instead of antibiotics in the therapy of mastitis. The healthy intestinal microflora structure is crucial for the cows' health. Probiotics has a potential ability to cure mastitis in dairy cows. In addition it plays an essential role in maintaining the healthy state and mastitis prevention.