Transcriptome-based Screening and Validation of Key Enzyme Genes for Polygonatum Polysaccharide Metabolism / 中国实验方剂学杂志

ObjectiveTo screen and validate key enzyme genes affecting the polysaccharide content in different Polygonatum species and perform in-depth amino acid sequence analysis by transcriptomic analysis of P. zanlanscianense， P. kingianum， and P. cyrtonema rhizomes to enrich the transcriptome data of Polygonatum plants and provide references for polysaccharide biosynthesis mechanism and genetic improvement. MethodThe Polygonatum transcriptome was sequenced and analyzed using the Illumina NovaSeq high-throughput sequencing platform， and the differences in the transcriptomes of the three Polygonatum species were compared and according to the annotations of Nr， Gene Ontology （GO）， and Kyoto Encyclopedia of Genes and Genomes （KEGG） databases. The key enzymes in the polysaccharide metabolism pathway were screened， and the expression of key enzyme genes was clustered and correlated with the polysaccharide content. Finally， Real-time polymerase chain reaction （Real-time PCR） was performed to validate the eight key enzyme genes， and the key genes of polysaccharide biosynthesis were further screened for homologous gene sequence analysis in combination with sequencing results， followed by constructing phylogenetic trees， predicting motifs， conserved structural domains， protein sequence isoelectric points， and molecular weights， and constructing 3D protein structures by using homology modeling method. ResultThe annotation of the Nr database revealed that three Polygonatum species had the highest gene homology with Asparagus officinalis. GO database annotation results showed that three Polygonatum species differed significantly in binding， catalytic activity， metabolic processes， and cellular components， while the KEGG pathway annotation results indicated that three Polygonatum species differed significantly in the starch and sucrose metabolic pathway and galactose metabolic pathway. According to clustering analysis， correlation analysis， Real-time PCR， expression profiles， and structural and functional predictions of amino acid sequences， the key enzyme significantly affecting the polysaccharide content in different Polygonatum species was inferred to be β-fructofuranosidase （sacA）. ConclusionSacA may be the main influencing factor for the difference in polysaccharide content of Polygonatum， and is also an important reason why Polygonatum polysaccharides are mainly fructans.

Pharmacokinetics and tissue distribution of four alkaloids in Ermiao Pills and Sanmiao Pills in normal and arthritic model rats / 中国中药杂志

This work aimed to investigate the differences of pharmacokinetics and tissue distribution of four alkaloids in Ermiao Pills and Sanmiao Pills in normal and arthritic model rats. The rat model of arthritis was established by injecting Freund's complete adjuvant, and ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) in the positive ion multiple reaction monitoring(MRM) mode was used for the determination of four alkaloids in plasma and tissues of normal and arthritic rats after administration of Ermiao Pills and Sanmiao Pills, respectively. The differences in pharmacokinetics and tissue distribution of the four active components were compared, and the effect of Achyranthis Bidentatae Radix on the major components of Sanmiao Pills was explored. This study established an UPLC-MS/MS for simultaneous determination of four alkaloids, and the specificity, linearity, accuracy, precision, and stability of this method all met the requirements. Pharmacokinetics study found that as compared with normal rats, the AUC and C_(max) of phellodendrine, magnoflorine, berberine and palmatine in model rats were significantly decreased after administration of Ermiao Pills, the clearance rate CL/F was significantly increased, and the distribution and tissue/plasma concentration ratio of the four alkaloids in the liver, kidney, and joint were significantly reduced. Achyranthis Bidentatae Radix increased the AUC of phellodendrine, berberine, and palmatine, reduced the clearance rate, and significantly increased the distribution of the four alkaloids in the liver, kidney, and joints in arthritic rats. However, it had no significant effect on the pharmacokinetics and tissue distribution of the four alkaloids in normal rats. These results suggest that Achyranthis Bidentatae Radix may play a guiding role in meridian through increasing the tissue distribution of effective components in Sanmiao Pills under arthritis states.

Effects and molecular mechanism of exogenous L-carnitine on excessive endoplasmic reticulum stress-mediated hepatic pyroptosis in severely scald rats / 中华烧伤杂志

Objective: To investigate the effects and molecular mechanism of exogenous L-carnitine on hepatic pyroptosis mediated by excessive endoplasmic reticulum stress in severely scald rats. Methods: The experimental research method was adopted. According to the random number table (the same group method below), fifteen female Sprague Dawley rats aged 6-8 weeks were divided into sham-injury group, scald alone group, and scald+carnitine group (with 5 rats in each group), and full-thickness scald of 30% total body surface area were made on the back of rats in scald alone group and scald+carnitine group, and rats in scald+carnitine group were additionally given intraperitoneal injection of L-carnitine. At post injury hour (PIH) 72, The levels of aspartate aminotransferase (AST) and alanine dehydrogenase (ALT) of biochemical indicators of liver injury were detected by automatic biochemical analyzer with the sample number of 5. At PIH 72, liver tissue damage was detected by hematoxylin-eosin staining. At PIH 72, The mRNA levels of nucleotide-binding oligomerization domain-containing protein-like receptor family pyrin domain containing 3 (NLRP3), cysteine aspartic acid specific protease 1 (caspase-1), gasderminD (GSDMD), and interleukin 1β(IL-1β) in liver tissue as pyroptosis-related markers and glucose regulatory protein 78 (GRP78) and CCAAT/enhancer binding protein homologous protein (CHOP) in liver tissue as endoplasmic reticulum stress-related markers were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR). Protein expression levels of GRP78, CHOP, NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1β in liver tissue were detected by Western blotting, and the sample numbers were all 5. HepG2 cells as human liver cancer cells were divided into dimethyl sulfoxide (DMSO) group, 0.1 μmol/L tunicamycin (TM) group, 0.2 μmol/L TM group, 0.4 μmol/L TM group, and 0.8 μmol/L TM group and were treated accordingly. After 24 h of culture, cell viability was detected by cell counting kit 8, and the intervention concentration of TM was screened, and the sample number was 5. HepG2 cells were divided into DMSO group, TM alone group, and TM+carnitine group, and treated accordingly. After 24 h of culture, the protein expression levels of GRP78, CHOP, NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1β in cells were detected by Western blotting, and the sample numbers were all 3. Data were statistically analyzed with one-way analysis of variance and least significant difference-t test. Results: At PIH 72, the AST and ALT levels of serum in scald alone group were (640±22) and (157±8) U/L, which were significantly higher than (106±13) and (42±6) U/L in sham-injury group, respectively, with t values of -46.78 and -25.98, respectively, P<0.01. The AST and ALT levels of serum in scald+carnitine group were (519±50) and (121±10) U/L, which were significantly lower than those in scald alone group, respectively, with t values of 4.93 and 6.06, respectively, P<0.01. At PIH 72, the morphology of liver tissue of rats in sham-injury group were basically normal with no obvious inflammatory cell infiltration; compared with those in sham-injury group, the liver tissue of rats in scald alone group showed a large number of inflammatory cell infiltration and disturbed cell arrangement; compared with that in scald alone group, the liver tissue of rats in scald+carnitine group showed a small amount of inflammatory cell infiltration. At PIH 72, the mRNA expression on levels of NLRP3, caspase-1, GSDMD, and IL-1β in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 34.42, 41.93, 30.17, and 15.68, respectively, P<0.01); the mRNA levels of NLRP3, caspase-1, GSDMD, and IL-1β in liver tissue of rats in scald+carnitine group were significantly lower than those in scald alone group (with t values of 34.40, 37.20, 19.95, and 7.88, respectively, P<0.01). At PIH 72, the protein expression levels of NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1β in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 12.28, 26.92, 5.20, 10.02, and 24.78, respectively, P<0.01); compared with those in scald alone group, the protein expression levels of NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1β in liver tissue of rats in scald+carnitine group were significantly decreased (with t values of 10.99, 27.96, 12.69, 8.96, and 12.27, respectively, P<0.01). At PIH 72, the mRNA levels of GRP78 and CHOP in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 21.00 and 16.52, respectively, P<0.01), and the mRNA levels of GRP78 and CHOP in liver tissue of rats in scald+carnitine group were significantly lower than those in scald alone group (with t values of 8.92 and 8.21, respectively, P<0.01); the protein expression levels of GRP78 and CHOP in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 22.50 and 14.29, respectively, P<0.01), and the protein expression levels of GRP78 and CHOP in liver tissue of rats in scald+carnitine group were significantly lower than those in scald alone group (with t values of 14.29 and 5.33 respectively, P<0.01). After 24 h of culture, the cell survival rates of 0.1 μmol/L TM group, 0.2 μmol/L TM group, 0.4 μmol/L TM group, and 0.8 μmol/L TM group were significantly decreased than that in DMSO group (with t values of 4.90, 9.35, 18.64, and 25.09, respectively, P<0.01). Then 0.8 μmol/L was selected as the intervention concentration of TM. After 24 h of culture, compared with that in DMSO group, the protein expression levels of GRP78 and CHOP in cells in TM alone group were significantly increased (with t values of 10.48 and 17.67, respectively, P<0.01), and the protein expression levels of GRP78 and CHOP in TM+carnitine group were significantly lower than those in TM alone group (with t values of 8.08 and 13.23, respectively, P<0.05 or P<0.01). After 24 h of culture, compared with those in DMSO group, the protein expression levels of NLRP3 and GSDMD-N in cells in TM alone group were significantly increased (with t values of 13.44 and 27.51, respectively, P<0.01), but the protein expression levels of caspase-1, caspase-1/p20, and cleaved IL-1β in cells were not significantly changed (P>0.05); compared with that in TM alone group, the protein expression levels of NLRP3 and GSDMD-N in cells in TM+carnitine group were significantly decreased (with t values of 20.49 and 21.95, respectively, P<0.01), but the protein expression levels of caspase-1, caspase-1/p20, and cleaved IL-1β in cells were not significantly changed (P>0.05). Conclusions: In severely scald rats, exogenous L-carnitine may play a protective role against liver injury by inhibiting the pathways related to excessive endoplasmic reticulum stress-mediated pyroptosis.

Clinical Characteristics and Prognostic Factors of Primary FL Patients with Grade 3 or Large B Cell Transformation / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical characteristics and prognostic factors of primary follicular lymphoma (FL) patients with grade 3 or large B cell transformation, so as to provide more reference for the subsequent clinical diagnosis and treatment.@*METHODS@#Forty-seven primary FL patients with grade 3 or large B cell transformation from March 2010 to March 2018 were selected, the clinical characteristics and survival of patients were analyzed. Cox regression model were used to evaluate the related prognostic factors.@*RESULTS@#The cumulative progression-free survival rate and cumulative overall survival rate of 47 patients in 3-year follow-up reached to 55.32% (26/47) and 80.85% (38/47) respectively. There were significant differences in cumulative progression-free survival rate and cumulative overall survival rate among different subgroups of IPI, FLIPI-1 and FLIPI-2 in 3-year follow-up (P3 cm lymph node-involved site number≥3, extranodal lesion site number≥2, IPI score=2-3, FLIPI-1 score and FLIPI-2 score≥3 were the risk factors for progression-free survival (P<0.05); LDH≥240 U/ml, IPI score=2-3 and FLIPI-2 score≥3 were risk factors for overall survival (P<0.05). Cox regression model multivariate analysis showed that IPI score=2-3 was the independent risk factor for progression-free survival and overall survival (P<0.05). FLIPI-2 score≥3 was the independent risk factor for overall survival (P<0.05).@*CONCLUSION@#Primary FL patients with grade 3 or large B cell transformation by using the existing treatment regimen might be possibly curable, and the current treatment strategies and IPI score can be used to predict the clinical prognosis of patients.

Predictive Value of Pancreatic Steatosis for Severity of Coronary Atherosclerosis in Patients with Type 2 Diabetes Mellitus / 中国医学科学院学报

To investigate the association of pancreatic steatosis with coronary atherosclerosis in patients with type 2 diabetes mellitus (T2DM). Patients with T2DM who underwent coronary computed tomography angiography(CCTA)in our center due to chest pain were enrolled from January 2016 to February 2019. According to the CCTA findings,patients were divided into normal group,mild-to-moderate coronary atherosclerosis group and severe coronary atherosclerosis group. CT attenuation of pancreas and spleen was measured on abdominal non-enhanced CT,and the CT attenuation indexes including the difference between pancreatic and splenic attenuation (P-S) and the ratio of pancreas-to-spleen attenuation (P/S) were calculated. Analysis of variance or Kruskal-Wallis rank test were used to assess differences among each group. Logistic regression analysis was used to analyze the risk factors of severe coronary stenosis. The accuracy of P/S in predicting severe coronary artery stenosis was assessed by receiver operator characteristic (ROC) curve analysis. A total of 173 consecutive T2DM patients were enrolled. These patients included 27 patients with normal coronary artery (15.6%),124 patients with mild to moderate stenosis (71.7%),and 22 patients with severe stenosis (12.7%). There were significant differences in CT attenuation of pancreas (=11.543,=0.003),P-S (=11.152,=0.004) and P/S (=11.327,=0.004) among normal coronary artery group,mild and moderate stenosis group,and severe stenosis group. The CT attenuation of pancreatic head,body,and tail significantly differed in patients with coronary artery stenosis (=14.737,=0.001). After adjusting for confounding factors,multiple Logistic regression showed that P/S (=0.062,95%=0.008-0.487,=0.008) was still significantly associated with the severe coronary artery stenosis. The area under the ROC curve of P/S for the diagnosis of severe coronary artery stenosis was 0.701,and the optimal cutoff point was 0.660. CT attenuation of pancreas and CT attenuation indexes are associated with the severity of coronary stenosis in T2DM patients,suggesting that pancreatic steatosis may be used as one of the indicators for predicting severe coronary artery stenosis.

Effect of the activation of P2X7 receptor on D-serine release in ventrolateral periaqueductal gray in rats with bone cancer pain / 医学研究生学报

Objective　The activation of P2X7 receptor in ventrolateral periaqueductal gray (vlPAG) is involved in the formation and maintenance of bone cancer pain (BCP). This study will establish a rat model of BCP and observe the effect of the activation of P2X7 receptor in vlPAG on D-serine level through brain microdialysis combined with ELISA.　Methods　Forty-two female SD rats were divided into four groups by random number table: normal control group (n=12), sham group (n=12), BCP group (n=12) and P2X7 receptor antagonist group (n= 6). The model of metastatic BCP in the tibias of the rats was established in the BCP group, and 20μL of RPMI-1640 medium cell suspension containing SHZ-88 breast cancer cells was injected (1×107 cancer cells/0.5 mL). The sham group was injected with treated cancer cells of the same volume (SHZ-88 breast cancer cells were kept in boiling water at 90 ℃ for 20 min), and the rest of the operation was the same as the BCP group. The normal control group received no treatment. The P2X7 receptor antagonist group was treated the same as the BCP group, except that the P2X7 receptor-specific antagonist A-438079 was added to the perfusion solution. The thermal pain threshold and mechanical pain threshold were detected at the same time in the normal control group, the sham group and the BCP group. The positive expression of P2X7 receptor in vlPAG of rats was detected by immunohistochemistry in each group in 21 days. The changes of D-serine in vlPAG dialysate were detected by ELISA in each group.　Results　The mechanical pain threshold and thermal pain threshold of the rats in BCP group on Day 5, 7, 10, 14, 18 and 21 were lower than those of the normal control group and sham group (P<0.01). The positive expression of P2X7 was scattered in vlPAG in normal control group and sham group. The number of P2X7 receptor positive cells in the BCP group was significantly higher than that in the control group and sham group (P<0.01). The content of D-serine in vlPAG of the rats in BCP group [(220.28±63.38)ng/mL] was significantly higher than that in the control group [(148.09±46.89)ng/mL] and the sham group [(147.32±51.44)ng/mL] (P<0.05). The content of D-serine in vlPAG [(134.20±41.77)ng/mL] in P2X7 receptor antagonist group was significantly lower than that in BCP group (P<0.05).　Conclusion　The activation of the P2X7 receptor in ventrolateral periaqueductal gray promotes D-serine release and participates in the mechanisms of BCP in rats .

Distribution characteristics of asthma in preschool children and the protective effect of breastfeeding / 中国学校卫生

Objective@#To clarify the distribution of asthma in preschool children aged 3-6 years old and the protective effect of breastfeeding on asthma in three provinces in the middle and lower reaches of the Yangtze river in China.@*Methods@#The parents of 27 200 preschoolers chosen from kindergartens in 11 cities of Hubei, Jiangsu and Anhui Province were investigated with children’s asthma and breastfeeding during October to November in 2017.@*Results@#The asthma detection rate for preschoolers was 4.0%,5.0% for boys and 2.9% for girls. The asthma detection rate for children aged 3,4,5,and 6 years old was 3.2%,3.7%,4.4%,and 4.9%. 3 448 (12.7%) and 7 630 (28.1%) preschoolers were never breastfed or mainly breastfed. The rate of exclusive breastfeeding was 50.0% for 1 to 5 months and only 10.2% over 6 months. Meanwhile,there was a significant protective effect on asthma (OR=0.88,95%CI=0.77-0.89) by exclusive breastfeeding for 1-5 months,which was still statistically significant after adjusting for confounding factors. After adjusting for confounding factors,exclusive breastfeeding over 6 month have significant effected on preschool children aged 4 years old without allergic conjunctivitis history (OR=0.63,95%CI=0.40-0.99). Furthermore,exclusive breastfeeding over 6 month had important effect on preschool children aged 4 years old with eczema history (OR=0.52,95%CI=0.27-0.99).@*Conclusion@#Exclusive breastfeeding has protective effect on asthma in preschool children. Exclusive breastfeeding over 6 months even has better protective effect on asthma for the 4-years-old children without the history of allergic conjunctivitis and eczema.

Effect of MiRNA-145 on Apoptosis of Leukemia HuT 78 Cells and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect and mechanism of miRNA-145 on leukemic cell apoptosis.@*METHODS@#After transfection of miRNA-145 mimic and negative control mimic in leukemia cells by Lipofectamine 2000 liposome, the MTT assay was used to detect the effect of miRNA-145 on cell proliferation. Flow cytometry was used to detect the effect of miRNA-145 on cell cycle and apoptosis. Western blotting assay was used to detect the expression levels of BCL-2, CDK6, Cyclin D1, BAX, PI3K p-PI3K, p-AKT and AKT.@*RESULTS@#The relative level of microRNA in HuT 78 cells transfected with miRNA-145 was 2.3±02, which was significantly higher than that in blank control group and miRNA-NC group (P＜0.05). MTT assay showed that the proliferation level of HuT 78 cells transfected with miRNA-145 mimic was significantly lower than that of blank control and miRNA-NC group (P＜0.05). Flow cytometry showed that the cells at G/G, S and G2/M phase of HuT 78 cells were significantly decreased after transfection with miRNA-145 mimic (P＜0.05). Annexin V/PI double staining assay showed that the apoptosis rate of HuT 78 cells was 17.6%±3.4%，which was significantly higher than that in blank control group and miRNA-NC group (P＜0.05). Western blot showed that the expression levels of BCL-2, CDK6 and Cyclin D1 in HuT 78 cells were significantly lower than those in blank control and miRNA-NC group (P＜0.05), and BAX expression in HuT 78 cells was significantly higher than that in blank control and miRNA-NC group (P＜0.05). Western blot showed that expression of PI3K, p-PI3K, AKT and p-AKT in HuT 78 cells transfected with miRNA-145 mimic were significantly lower than that in blank control and miRNA-NC group (P＜0.05).@*CONCLUSION@#Upregulation of miRNA-145 may inhibit the proliferation of leukemia cells and promote the apoptosis, which may be related with the inhibition of PI3K/AKT signaling pathway.

Aislamiento y caracterización de una bacteria marina degradadora de testosterona: Vibrio sp. N3 / Isolation and characterization of a marine testosterone-degrading bacterium: Vibrio sp. N3

Steroids, including testosterone, estrone, 17β-estradiol, estriol and 17β-ethinyl estradiol, are harmful not only to the population dynamics of aquatic life forms but also to public health. In this study, a marine testosterone-degrading bacterium (strain N3) was isolated from Nanao Island in the South China Sea. In addition, the strain could also use 17β-estradiol (E2), 17β-ethinyl estradiol (EE2), estriol (E3) or cholesterol as a sole carbon source. According to the 16S rRNA gene sequence analysis, strain N3 was identified as Vibrio sp. Further characterization showed that the strain is aerobic, gram-negative, and mobile and exhibits resistance to ampicillin, carbenicillin, penicillin and spectinomycin. For enhancing its capacity of testosterone degradation, the Plackett-Burman factorial design and the central composite design were used to optimize the culture condition. Under optimal conditions, 92% of testosterone was degraded by Vibrio sp. N3 in 48 h.

Los esferoides-que incluyen la testosterona, la estrona, el 17 β-estradiol, el estriol y el 17 p-etinilestradiol-son nocivos no solo para la población dinámica de las formas de vida acuática, sino también para la salud pública. En este estudio se aisló una bacteria marina degradadora de testosterona de la isla de Nanao, en el Mar del Sur de China, a la que se denominó cepa N3. Se determinó que esta cepa también podría usar 17 β-estradiol (E2), 17 p-etinilestradiol (EE2), estriol (E3) o colesterol como únicas fuentes de carbono. De acuerdo con el análisis de la secuencia del gen 16S rRNA, la cepa N3 se identificó como Vibrio sp. La caracterización adicional mostró que dicha bacteria es un organismo aerobio, gram negativo y móvil, y que presenta resistencia a ampicilina, carbenicilina, penicilina y espectinomicina. Para optimizar la condición de cultivo en relación con su capacidad de degradar la testosterona, se utilizaron el diseño factorial Plackett-Burman y el diseno compuesto central. En condiciones óptimas, el 92% de la testosterona fue degradada por Vibrio sp. N3 en 48 h.

The effect of activation of P2X7 receptor on Ca2+ level in lateral midbrain periaqueductal gray neuron / 医学研究生学报

Objective The main intracellular signal of P2X7 receptor activation is the increasing of Ca2+, which then presents the diversity of its physiological and pathological functions through multiple intracellular signal transduction. To observe the effect of activation of P2X7 receptor on intracellular calcium（Ca2+）level in lateral midbrain periaqueductal gray (lPAG) neurons of primary cultured rat. Methods The primary cultured lPAG neurons were randomly divided into 4 groups: control group(no drug added), only for control; BzATP group(100 μmol/L); A-740003+ BzATP group(incubate with 100 nmol/L A-740003 for 10 min, then add 10 μmol/L ofBzATP); BzATP control group(add in Ca2+-free solution for 20 min, then add BzATP). The incubation solution of control group, BzATP group and A-740003+ BzATP group are DMEM/F12 medium, and the BzATP control group is Ca2+-free. The laser scanning confocal microscopy (LSCM) was used to detect : the changes of cultured neuron Ca2+ levels by different concentrations of BzATP; the effects of A-740003 and Ca2+-free medium preincubation on BzATP-induced Ca2+ level alterations in cultured neurons. Results BzATP dose-dependently increased the Ca2+ levels in cultured lPAG neurons; A-740003 and Ca2+-free medium inhibited the BzATP-induced increasing of Ca2+ level in cultured lPAG neurons. LSCM showed: The intracellular calcium fluorescence insensity(2.48±1.05) in the BzATP group was significantly higher than that in the blank control group, BzATP control group and A-740003+ BzATP group[(1.12±0.03), (1.09±0.03), (1.14±0.08)](P<0.01). Conclusion The activation of P2X7 receptor can increase the level of lPAG neurons Ca2+ , and is associated with the extracellular Ca2+ influx.

Pharmacokinetics of eight constituents in rat plasma after oral administration of Modified Xiaochaihu Granules for gastric ulcer based on UPLC-MS/MS / 中国中药杂志

An ultra-performance liquid chromatography-tandem mass spectrometric method (UPLC-MS/MS) was developed in this study to simultaneously determine the contents of eight effective constituents in rat plasma, including baicalin, wogonoside, baicalein, liquiritin, glycyrrhizic acid, berberine hydrochloride, saikosaponin a and saikosaponin d in plasma of gastric ulcer rats, and investigate the pharmacokinetics of Modified Xiaochaihu Granules. Chromatographic separation was conducted on Zorbax SB-C₁₈ column (2.1 mm×100 mm, 1.8 μm) with acetonitrile -0.1% formic acid aqueous solution as the mobile phase for gradient elution, at a flow rate of 0.4 mL·min⁻¹ and column temperature of 40 °C. Detection was performed in the multiple reaction monitoring (MRM) mode with ESI ion source. All calibration curves showed good linearity (>0.996) over a wide concentration range for all constituents. RSDs of intra-day and inter-day precision were all within 15% and the extraction recoveries of all the constituents were in the range of 81.92% to 104.8%. The time to peak (tmax) of these eight constituents was (2.69±2.02), (5.17±2.04), (0.25±0), (0.83±0.26), (0.92±0.20), (0.92±0.20), (0.58±0.20), and (0.083±0) h, respectively; the half-life (t1/2) of them was (7.85±0.34), (10.16±2.21), (6.79±0.21), (8.32±0.48), (11.05±1.78), (11.56±3.46), (15.30±1.84), and (5.54±1.91) h, respectively; the peak concentration (Cmax) of them was (55.02±1.67), (213.66±4.62), (62.61±0.69), (68.43±1.42), (62.22±0.39), (30.17±1.89), (61.79±4.81), and (38.02±1.75) μg·L⁻¹, respectively. This established method is simple and accurate with good repeatability and strong specificity, which could provide modern experimental basis for modified Xiaochaihu granules in clinical treatment of gastric ulcer.

Studies on serum pharmacochemistry of effective parts of modified Xiaochaihu Tang for treatment of gastric ulcer / 中国中药杂志

To conduct the studies on serum pharmacochemistry of effective parts of modified Xiaochaihu Tang for treatment of gastric ulcer by using chromatography-mass spectrometry. Absolute ethyl alcohol induced-gastric ulcer model of mouse was used in this study to investigate the pharmacology of modified Xiaochaihu Tang and its effective parts. Both groups could significantly decrease the absolute ethyl alcohol induced-ulcer. Gasphase-mass spectrometry (GC-MS) was used to detect chemical compositions of volatile fractions and the drug components after gastric administration. A total of 63 compounds were identified in extracts, accounting for more than 93% of the all volatile oil, including 23.51% α-curcumene, 11.96% zingiberene, 9.56% curzerene, 6.54% β-sesquinene, 4.77% camphene, and 6 prototype components were also detected in serum for gastric ulcer model. In liquid chromatography-mass spectrometry (LC-MS), a total of 17 compounds were identified in extracts, 6 prototype components and 2 metabolites (3,5-O-feruloylquinic acid and palmatine) were obtained in serum. In a conclusion, this study provides an important scientific basis for identifying the active ingredients in modified Xiaochaihu Tang, and also helps to reveal the pharmacological effect of modified Xiaochaihu Tang for treatment of gastric ulcer.

Effects of Celastrol on Accelerated Healing of Skin Wounds of Diabetic Ulceration in Rats and Mechanism Revealed by Nuclear Magnetic Resonance-based Metabonomics / 分析化学

The experimental SD rats were randomly divided into normal control group (Con group),diabetic ulcer model group (DM group) and Celastrol group (Cel group).Except the control group,diabetic ulceration rat models were established by intraperitoneal injection of streptozotocin along with skin scald.And then,each group was treated by spraying the saline solution on the affected skin with (Cel group) or without (Con group and DM group) Cel (q.d.×14 d).Nuclear magnetic resonance (NMR)-based metabonomic analysis was applied to detect metabolic characteristics,accompanied by healing rate calculation and HE and Masson staining to study therapeutic effect of celastrol on accelerated healing of skin wounds of diabetic ulceration rats,which could be used to elucidate therapeutic effects of celastrol on the rat diabetic ulceration and its mechanism.The results showed that celastrol could induce epithelial regeneration of the rat ulcer wound,regulate the infiltration of inflammatory cells and the distribution of collagen fibers,and promote the healing of the ulcer wound.About 20 endogenous potential differential metabolites were screened and identified by partial least square analysis.Metabolic pathway analysis was carried out to show that celastrol can significantly recovery the level of the tricarboxylic acid cycle,promote its energy supply,accelerate the protein synthesis,improve mitochondrial dysfunction and oxidative stress,and accelerate the self-repair ability of skin tissue.Celastrol can promote the healing of ulcers skins of the diabetic rats,which contribute to experimental basis of the drugs for the treatment of diabetic ulcers.

Advances in studies on cancerous pain-related ion channels / 医学研究生学报

With the improvement of tumor diagnosis and treatment, the survival time in cancer patients has been gradually prolonged, but the complications such as severe pain are often found. Due to the unrevealed mechanisms of cancer pain, clinical analgesic treatments are unsatisfied needs , and the cancerous pain seriously affected the life quality of icancer patients. Ion channels are the transmembrane glycoproteins which produce and transmit electrical signals when activated, and involved in the regulation of a variety of physiological and pathological processes. Alterations in the ion channel function may play an important role in the formation and maintenance of cancer pain. In this review, the research developments of several major ion channels in the mechanisms of cancer pain in recent years are summarized, which can provide some useful references for the study of ion channel and related drug development of cancer pain.

Study on the Influence of Medical Insurance Differentiation Reimbursement Policy on Patients' Flow / 中国卫生经济

Objective:The elasticity and correlation analysis were applied to analyze the influence of medical insurance differentiation reimbursement policy to guide the flow of patients,as well as the implementation of the policy of hierarchical medical system to find the problem,and then make policy recommendations for hierarchical diagnosis and health care.Methods:Yanqing District(Beijing),Mishan and Hulin(Heilongjiang) and Longxi County(Gansu) were selected as four research sites to collect each site's New Rural Cooperative Medical System(NCMS) annual data for quantitative analysis.The sample areas' NCMS annual data (from 2013 to 2015)were collected for correlation analysis and elasticity calculation by SPSS 23.0.Results:(1) Each site patients' elasticity of income and medical service price was low,the elasticity of outpatient volume-hospital service price was high.(2) Patients' behavior of choosing medical institutions to seek medical advice had low correlation with their income level and medical service prices,which had formed stable medical habits.Conclusion:Medicare differentiated reimbursement policy had a certain effect on guiding the flow of patients,but the effect was not obvious in the county.The main obstacles to implement basic initial diagnosis were the weak capacity of grassroots medical institutions and policy support was not in place.It was suggested that the health insurance differentiation reimbursement policy should be combined with the disease restriction policy,strengthen the grassroots capacity building,establish and improve the supporting policy.

Application of highly selective renal artery occlusion in laparoscopic partial nephrectomy / 中国内镜杂志

Objective To compare the feasibility and safety of laparoscopic partial renal segmental artery occlusion and renal artery trunk interruption for partial nephrectomy. Methods We reviewed medical records of 65 patients with stage T1 renal tumor from October 2013 to February 2017. Among them, 29 cases underwent partial nephrectomy with high selective segmental renal artery occlusion, and 36 cases underwent partial nephrectomy with renal artery trunk occlusion. Then compare the preoperative and postoperative creatinine changes, intraoperative blood loss, warm ischemia time, operation time, hospitalization time and positive margins of the two methods. Results In all the patients, the procedures were done without conversion to open surgeries. The operation time of the renal artery branch block group is shorter than that of the main renal artery block, but the preoperative and postoperative creatinine changes are smaller than those of the main renal artery occlusion group. There was no significant difference between the two groups in intraoperative blood loss, warm ischemia time and positive margins (P > 0.05); There was no significant difference between the two groups in age, body mass index (BMI) and R.E.N.A.L score of renal tumor (P > 0.05). Conclusion For the stage T1 renal tumor, the selective renal artery branch interruption technique has a longer operation time, but the renal tissue has a small ischemic zone and has little effect on the kidney.

Effects of miR-34 on migration and invasion of prostate cancer cells by MSR1 protein / 中国免疫学杂志

Objective:To investigate the effect of miR-34a on the migration and invasion of prostatic cell line through MSR1.Methods:Western blot was used to detect the expression of MSR1 in prostate cancer cell lines.qPCR was used to detect the ex-pression of miR-34a in prostate cancer cell lines.The correlation between miR-34a and MSR1 was examined by double luciferase as-say.Transwell invasion assay was used to detect the effect of miR-34a and MSR1 on the invasion ability of prostate cancer cells.The effects of miR-34a and MSR1 on the fine migration ability of prostate cancer were examined by scratch healing.Results:The expression level of MSR1 in PC3 cells was relatively high by Western blot.The expression level of miR-34a in PC3 cells was relatively low by qPCR.Double luciferase assay showed a direct binding site between miR-34a and MSR1,MiR-34a could regulate the expression level of MSR1,Scaffold Healing Test and Transwell Invasion Test were used to detect the migration and invasion of PC3 cells after overexpressing miR-34a.Overexpression of MSR1 could reverse the inhibitory effect of miR-34a on the migration and invasion of PC3 cells.Conclusion:miR-34a can regulate the migration and invasion of prostate cancer cells through MSR1 protein.

Activation of spinal P2X7 receptor and analgesic effect of tramadol on neuropathic pain in rats / 医学研究生学报

Objective The mechanisms underlying neuropathic pain are complicated and the clinical effect of analgesia therap on this condition is not quite satisfactory.In this study, we observed the analgesic effects of the different doses of tramadol (T ) on neuropathic pain in rats and explored its action mechanisms . Mehtods The model of chronic sciatic nerve constriction injury (CC)I was established in male SD rats.The rats were randomly divided into five groups , sham operation ,CCI model control, low-dose T, mediumdose T, and high-dose T ,those in the latter three groups injected intraperitoneally with T at 5 , 15,and 25mg /kg qd ,respectively ,from the 7th to the 14th day after modeling.The mechanical and thermal pain thresholds of the nerve -injured hindleg were measured pre-operatively and at1 ,5 , 7, 10, 12and 14 days post-operatively.At 14 days after modeling, the expression of the P2X7receptor in the spinal dorsal horn was detected by immunohistochemistry and Western blot . Results At 5, 7,10 , 12 and 14 days after modeling,the mechanical pain threshold values were significantly decreased in the rats of the CCI model control group ([34.97±3.86 ],[34.06 ±3.79], [ 33.27±3.65], [29.03±3 . 54], and [17.90±2.34] g) and high-dose Tgroup([ 34.87±3.85], [33.47±3.66],[34 .50±3. 78 ], [29.43±3.64], and [18.63±2.42] g) as compared with the animals of the sham operation group ([39.73±5.55],[39.50±5.51], [40.97±5. 58], [41.87±5.60], and [42.97±6.75] g) (all P<0.01), and so were the thermal pain threshold valuesin the CCI model control group ([35 .21±3.94], [35.16±3.80], [29.74±2. 76], [ 20. 47±2.16], and[12.08±1.48] s) and high -dose T group ([35.76±3.76], [33.27±3.52], [31.22±3.05], [19.41±2.08], and [10.35±1.34] s) in comparison with the shamoperation group ([39.69±4.86], [39.21±4.82], [39.42±5.08], [41.17±4.88], and [42.53±5.12] s) (all P<0. 01 ).The number of the P2X7receptor positive cells and the ROD value of the P 2X7 receptor protein in the spinal dorsal horn were remarkably higher in the CCI model control than in the sham operation , low-dose T and medium-dose T groups (all P<0.01). Conclusion Intraperitoneal injection of low-dose tramadol has an analgesic effect on neuropathic pain in rats , which may be related to its decreasing effect on theexpression of the P2X7 receptor in the spinal dorsal horn.

Classification and treatment of syndesmotic injury / 中国骨伤

The distal tibiofibular syndesmosis is a critical structure in maintaining the ankle stability. Syndesmotic injuries are usually associated with ankle fractures and high fibula fractures. Non-isolated and partially isolated syndesmotic injuries are involved in unstable injuries, which need to operative treatment. Partially isolated syndesmotic injuries belong to stable injuries, which should be treated with non-operative management. It is becoming clear that early fixation and stabilization for unstable injuries are probably better than non-treatment or delayed treatment. It still remains without consensus of accurately defining stable from unstable injuries and sufficiently differentiating between acute and chronic injuries. Because of stability, fixation type, and duration, the clinical efficacy is different. Screw fixation is a gold standard treatment of syndesmotic injury. However, it remains controversial that whether removal of the syndesmotic screw is required and effect of the level of syndesmotic screw insertion, limited micro-movement is one of disadvantages of screw fixation. Micro-movement of the distal tibiofibular syndesmosis has been paid more and more attention. Dynamic fixation is a viable alternative to the static fixation device, with lower re-operation rates and less complications, which has obtained a great short-term clinical efficacy. However, further long-term studies should be carried out to confirm this clinical efficacy. Optimized treatment strategies considering stability of syndesmotic injury, duration, and fixation type can help to improve clinical efficacy.

Behavioral logic of community health service institutions in the background of the new healthcare reform based on the grounded theory / 中国卫生政策研究

Community health service institutions are the basic tentacles of medical services which directly indicates their relation to the quality and experience of health service.This paper focuses on the behavioral logic of community health service institutions under the background of new medical reform, and looks forward to a perspective on the grassroots practice caused by the reform of medical system through the behavioral analysis of the community health service institutions.The behavioral logic features of the community health service institutions were constructed based on the interview data Nanjing City using the grounded theory.It was found that the realization of public welfare orientation of the aforesaid institutions needs to be completed under the internal and external coordination.The degree of coordination among the interests, orientation, incentives and resource elements will determine the behavior orientation and basic operational practices in the community health service institutions.This study will provide a reference value for the cognition and optimization of healthcare reform policies.