OBJECTIVETo explore the current application and features of Aconite prescriptions with incompatible herbs in grade A class three hospitals in east China and central China through a clinical study and comparative analysis.

METHODClinical prescriptions containing Aconite with incompatible herbs were collected. Association rules were utilized to analyze the compatible features of these herbs.

RESULTThis analysis found that the frequently used incompatible herba; pairs are Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma, with the support rate of 44.45%, occupying nearly half of the surveyed prescriptions; Pinelliae Rhizoma is the most frequently used herb in the two areas, with support rate up to 76.24%. Among the top 10 herbal pairs in the support rate, except for Aconiti Lateralis Radix Praeparata and Pinelliae Rhizoma, the top 10 herbs in Central China were mostly for warming the middle jiao and tonifying qi, such as Zingiberis Rhizoma, Atractylodis Macrocephalae Rhizoma and Codonopsis Radix; Whereas those in east China were mostly for activating and nourishing blood, such as Angelicae Sinensis Radix, Chuanxiong Rhizoma, and Salviae Miltiorrhizae Radix et Rhizoma. Among the top 10 herbal pairs in the support rate, except for Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma, the core herbal pairs applied in central China were mainly for resolving phlegm and warming the middle jiao, such as Pinelliae Rhizoma-Glycyrrhizae Radix et Rhizoma, Pinelliae Rhizoma-Zingiberis Rhizoma; Whereas those in east China were principally for activating blood and tonifying qi, like Atractylodis Macrocephalae Rhizoma and Pinelliae Rhizoma, Angelicae Sinensis Radix and Pinelliae Rhizoma. Among the core herbal groups in the two areas, the most frequently used herbal groups in the two areas are Aconiti Lateralis Radix Praeparata, Glycyrrhizae Radix et Rhizoma and Pinelliae Rhizoma with the support rate of 59.73%, accounting for the highest proportion among all of herbal groups.

CONCLUSIONThere are the combined clinical application of Aconite with incompatible herbs, mostly with Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma, but with differences in the combined application in east China and central China.

Aconitum ; chemistry ; Drug Incompatibility ; Drug Prescriptions ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Pinellia ; chemistry ; Sophora ; chemistry

Objective To investigate the predictive value of preoperative serum human epididymis protein 4 (HE4) and CA125 in patients with high risk of stage Ⅰ endometrial cancer (EC)and to identify the optimal cutoff values. Methods Clinical and pathological data of 231 patients with stage Ⅰ EC were included in this study. Patients were divided into high risk group (n=96) and low risk group (n=135). The preoperative serum levels of HE4 and CA125 were measured, and their correlations with clinical pathological features were analyzed. The ROC curves were generated to determine optimal cutoff values of HE4 and CA125 levels with the maximum Youden index for prediction of high risk EC. Results There were significant differences in serum levels of HE4 and CA125 between patients with different depths of myometrial invasion, with or without vascular invasion, with or without lower uterine segment involvement, with different diameters of tumor and different risk classifications of stage ⅠEC (P<0.05). There were significant differences in serum levels of HE4 between patients with different menopausal status, hypertension, pathological types, histological grading and the involvement of cervical endometrial glands (P<0.05). The preoperative serum levels of HE4 and CA125 were positive correlated (r=0.262, P<0.05). The AUC value of HE4 for diagnosing stageⅠEC was 0.794(95%CI:0.734-0.854),the cutoff value was 74 pmol/L, the sensitivity was 75.0%, specificity was 83.0%, positive predictive value was 75.8%, negative predictive value was 82.4% and the accuracy rate was 79.7%. The AUC value of CA125 for diagnosing stage Ⅰ EC was 0.696 (95%CI: 0.624-0.767), the cutoff value was 17 kU/L, the sensitivity was 56.3%, specificity was 85.9%, positive predictive value was 74.0%, negative predictive value was 73.4%and the accuracy rate was 73.6%, respectively. The AUC value of combination of both markers was 0.847 (95%CI: 0.796-0.899), the corresponding values were 95.8%, 77.0%, 74.8%, 96.3%and 84.8%, respectively. Conclusion The best cutoff values of HE4 and CA125 are 74 pmol/L and 17 kU/L for detecting high risk stageⅠEC. The combined detection is better than that of the single detection in sensitivity, negative predictive value and accuracy rate.

OBJECTIVE: To establish 2-dimensional electrophoresis (2-DE) maps of Helicobac-ter pylori in human gastritis, and gastric cancer, to identify the differentially expressed proteins,and to discuss the role of bacterial factor in pathogenesis. METHODS: The total proteins of Helicobacter pylori in human gastritis and gastric cancer were separated by immobilized pH gradient-based 2-DE. The differentially expressed proteins were screened by PDQuest analysis software and identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of flight mass spectrometry, and searched on database. RESULTS: A well-resolved and reproducible 2-DE pattern of Helicobacter pylori was obtained from patients with human gastritis and gastric cancer. Fourteen differentially expressed proteins were identified, including proteins related to anti-oxidation,molecular chape-rones and detoxification, enzymes related to metabolism,proteins related to cytoarchitecture,and proteins related to signal conduction. CONCLUSION A well-resolved and reproducible 2-DE pattern of Helicobacter pylori in human gastritis and gastric cancer is established and differentially expressed proteins from these 2 diseases are identified. The differentiation of protein expression may play an important role in the pathogenesis of gastric cancer.
Bacterial Proteins ; analysis ; Electrophoresis, Gel, Two-Dimensional ; Female ; Gastritis ; microbiology ; Helicobacter Infections ; microbiology ; Helicobacter pylori ; Humans ; Male ; Proteome ; analysis ; Proteomics ; methods ; Stomach Neoplasms ; microbiology

OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

Objective:To analyze the clinical prognostic factors of liposarcoma on the extremities and trunk, as well as to retrospectively analyze the effect of adjuvant radiotherapy and chemotherapy on liposarcoma of the extremities. Methods:Patients with liposarcoma of the extremities treated in our hospital from July 1, 2007 to December 31, 2012 were followed up. The relationship of clinical prognostic factors with gender, age, location, depth, and size of the tumors, as well as the histological grade and admission status, were statistically analyzed. The effects of adjuvant radiotherapy or chemotherapy on overall survival (OS) and disease free survival (DFS) were evaluated. Results:A total of 82 patients with extremity liposarcoma received surgery-based comprehensive treatment in our hospital. Of the total patients, 73 received a 24-month to 88-month satisfied follow-up;the median follow-up time was 47 months. The OS rate was 83.6%(61/73), and the DFS rate was 68.5%(50/73). Multivariate Cox regression analysis showed that the tumor location, histological grade, and admission status were the independent correlative factors influencing DFS, and the age and pathologic grading were the independent correlative factors influencing the OS. Kaplan-Meier survival analysis showed that radiation therapy can significantly improve the DFS and OS of the G2 and G3-grade liposarcoma (DFS:59.1 months vs. 28.4 months, P<0.01;OS:70.8 months vs. 55.1 months, P<0.05). Significant difference was not found in the effect of chemotherapy on OS and DFS. Conclusion:The prognosis of liposarcoma was significantly associated with the pathologic grades and subtypes. Auxiliary radiotherapy could improve the survival and prognosis of G2 and G3 liposarcoma of the extremities, but the role of chemotherapy in treating liposarcoma remained unclear.

OBJECTIVETo investigate the effects of the combination of musk and olibanum on the tight junction protein expressions in prostatic epithelial cells of normal and chronic prostatitis (CP) rats.

METHODSEighty male SD rats were randomly divided into 8 groups of equal number: normal control, normal musk, normal olibanum, normal musk + olibanum, CP model control, CP model musk, CP model olibanum, and CP model musk + olibanum. At 60 days after modeling, the rats in the control, musk, olibanum, and musk + olibanum groups were treated intragastrically with normal saline, musk (0.021 g per kg body weight per day), olibanum (1.05 g per kg body weight per day), or musk + olibanum respectively, all for 3 days. Then, all the rats were sacrificed and their prostate tissues harvested for detection of the expressions of the tight junction proteins Claudin-1, Claudin-3, Occludin, and ZO-1 in the prostatic epithelial cells by immunohistochemical staining.

RESULTSIn the CP models, only the expression of Claudin-1 was significantly increased. In the normal rats, the expression of Claudin-1 was remarkably upregulated after treated with musk (824.6 ± 393.3, P < 0.05), olibanum (982.0 ± 334.0, P < 0.05), and musk + olibanum (1088.1 ± 640.2, P < 0.01); that of Claudin-3 was elevated markedly by olibanum (1 009.5 ± 243.6, P < 0.05) and insignificantly by musk (597.5 ± 80.7), but the increasing effect of olibanum was reduced by musk + olibanum (678.4 ± 255.1). No statistically significant differences were found in the expression of Occludin among the rats treated with musk (693.0 ± 424.8), olibanum (732.1 ± 302.0), and musk + olibanum (560.2 ± 202.3), or in that of ZO-1 in the animals treated with musk (290.0 ± 166.8) and olibanum (419.7 ± 108.1), but the latter was markedly decreased in the musk + olibanum group (197.7 ± 98.2, P < 0.05). In the CP rat models, both the expressions of Claudin-1 (823.0 ± 100.1, P < 0.01) and Occludin (1160.0 ± 32.2, P < 0.05) were significantly increased. The expression of Claudin-1 was remarkably down-regulated by musk (764.9 ± 179.0), olibanum (468.4 ± 220.4), and musk + olibanum (335.1 ± 204.0) (all P < 0.05), but that of Claudin-3 up-regulated by musk (744.6 ± 94.5) and olibanum (700.1 ± 223.7) (both P < 0.05). The expression of Occludin was reduced by musk (615.0 ± 221.0), olibanum (749.6 ± 321.7), and musk + olibanum (505.8 ± 523.7), while that of ZO-1 increased by olibaum (443.2 ± 44.9) and decreased by musk + olibanum (213.5 ± 24.9, P < 0.05).

CONCLUSIONIn physiological and pathological conditions, the combination of musk and olibanum acts on the expressions of tight junction proteins in prostate epithelial cells in a selective and dual-targeting manner, promoting their permeability by down-regulating the expression of ZO-1 and maintaining their structural stability by regulating the expressions of Claudin-1, Claudin-3, and Occludin.

Animals ; Claudins ; metabolism ; Down-Regulation ; Epithelial Cells ; drug effects ; Fatty Acids, Monounsaturated ; chemistry ; Frankincense ; chemistry ; Male ; Occludin ; metabolism ; Prostate ; cytology ; Prostatitis ; Rats ; Rats, Sprague-Dawley ; Tight Junction Proteins ; metabolism ; Up-Regulation

OBJECTIVETo observe whether H. pylori inoculated by oral route could arrive in livers and cause liver inflammation as an independent etiological factor.

METHODSC57BL/6 mice were orally inoculated with H. pylori SS1 strains and fed for 8 months. H. pylori colonization and pathologic consequences were studied in the liver and gallbladder tissues of the mice; the blood, liver tissue and gastric mucosa were obtained and cultured for H. pylori growth; The bacterial DNA extracted from the liver, bile and blood was examined by nested PCR for H. pylori genes. 16S rRNA PCR amplicons were sequenced and compared with the sequencing results of 16S rRNA PCR amplicons of the bacteria cultured from gastric mucosa and the inoculated H. pylori SS1.

RESULTSThe bacterial DNA extracted from the liver, bile and blood of the infected mice was detected for H. pylori genes by nested PCR. Six of the 15 samples were positive (40%) in the liver, 6 of 10 samples in the bile (60%), and 2 of 10 samples in the blood (20%). Sequencing results of 16S rRNA PCR products of the livers showed 100% homogeneity when compared with the cultured H. pylori from gastric mucosa and inoculated H. pylori SS1. H. pylori was found in 4 liver tissues of the 15 infected mice (26.7%) and 6 in the gallbladders (40%). Infiltrations of lymphocyte cells along hepatic sinusoids and a lower degree infiltration around interlobular arteries and veins were observed; ballooning degeneration was also observed in some hepatocytes.

CONCLUSIONH. pylori inoculated by oral route could arrive in the liver and cause inflammation as an independent etiological factor. The routes which the microorganisms took to reach the livers may involve hematogenous and/or biliary system dissemination.

Animals ; Helicobacter Infections ; Helicobacter pylori ; pathogenicity ; Liver ; microbiology ; Male ; Mice ; Mice, Inbred C57BL ; Rats

OBJECTIVETo observe whether H. pylori administered orally in mice could arrive in their livers after a long-term infection, leading to active inflammation and even causing HCC as an independent etiological factor.

METHODSTwenty C57BL/6 mice were orally administered H. pylori SS1 and kept for 24 months (experimental group) along with 13 mice which served as blank controls (control group). H. pylori colonization and pathologic consequences were studied in the livers and gastric tissues of the mice. The bacterial DNA extracted from liver tissues was examined by nested PCR for H. pylori 16S rRNA genes. 16S rRNA PCR amplicons were sequenced and compared with sequencing results of 16S rRNA PCR amplicons of the bacteria cultured from gastric mucosa and compared with that of the inoculated H. pylori SS1.

RESULTSOf the 20 mice in the experimental group, H. pylori was found in the gastric mucosa of 12, and in 11 of them pathological gastric lesions were found, including one with gastric lymphoma. H. pylori were found in the livers of 7 mice. Liver lesions, one with mild inflammation, 3 with inflammation and fibrosis, 2 with inflammation, fibrosis and hepatocyte hyperplasia with atypia were found in 6 of them. No liver lesions were found in the mice of the control group. In the mice of the experimental group no liver lesions were found in those mice with no H. pylori in their gastric mucosae. Sequencing results of 16S rRNA PCR products of the liver showed 100% homogeneity with the cultured H. pylori from gastric mucosa and the administered H. pylori SS1.

CONCLUSIONTwo years after oral administration of H. pylori to C57BL/6 mice, gastric mucosal lesions and liver lesions, including inflammation, cirrhosis and hepatocyte hyperplasia with atypia were found in those animals.

Animals ; Disease Models, Animal ; Helicobacter Infections ; pathology ; Helicobacter pylori ; Liver ; microbiology ; pathology ; Male ; Mice ; Mice, Inbred C57BL

AIM To prepare cucurbitacin B nanosuspensions,and to investigate their in vivo pharmacokinetics.METHODS The nanosuspensions were prepared by high-pressure homogenization method.With stabilizer type,stabilizer-drug ratio and homogeneous frequency as influencing factors,particle size and PDI as evaluation indices,the formulation was optimized by single factor test,after which the solubility and stability were determined,and crystalline form analysis was performed.Eighteen rats were randomly assigned into three groups and given intragastric administration of the 0.5%CMC-Na suspensions of cucurbitacin B,physical mixture and cucurbitacin B nanosuspensions(10 mg/kg),respectively,after which blood collection was made at 0.5,1,2,3,4,8,10,12 h,UPLC-MS/MS was adopted in the plasma concentration determination of cucurbitacin B,and main pharmacokinetic parameters were calculated.RESULTS The optimal formulation was hydroxypropyl cellulose+sodium dodecyl sulfate(1 ∶ 1)as stabilizer,3 ∶ 1 for stabilizer-drug ratio,80 MPa for homogeneous pressure,and 12 times for homogeneous frequency,the average particle size,PDI and Zeta potential were 200 nm,0.140 and-32 mV,respectively.The nanosuspensions demonstrated obviously higher solubility than that of raw medicine and physical mixture,along with good stability within 6 months.Cucurbitacin B existed in the nanosuspensions in an amorphous state.Compared with raw medicine and physical mixture,the nanosuspensions displayed shortened tmax(P<0.01),prolonged t1/2(P<0.05,P<0.01),and increased Cmax,AUC0-t,AUC0-∞(P<0.01),whose relative bioavailability was enhanced to 4.32 times as compared with that of raw medicine.CONCLUSION Nanosuspensions can improve the dissolution rate and oral bioavailability of cucurbitacin B.

OBJECTIVETo study the effects of drug plasma of musk and olibanum (DP-M&O) on the release of inflammatory cytokines from monocytes and the expressions of the proteins associated with inflammation of prostatic or endothelial cells induced by prostate antigen (PAg) stimulation.

METHODSWe prepared DP-M&O using SD rats and monocytes and PAgs using BALB/c mice. We pre-treated the monocytes with DP-M&O at the gradient concentrations of 0, 2.5, 5, 10, and 20% for 1 hour, activated them with PAgs, and then cultured them for 96 hours, followed by detection of the release of inflammatory cytokines. We co-cultured the prostate RWPE-1 cells with the endothelial EA. hy926 cells, pre-treated them with the same gradient concentrations of DP-M&O as above for 1 hour, activated with PAgs, and cultured for 96 hours. Then we determined the expression levels of the proteins associated with inflammation of RWPE-1 and EA. hy926 cells by Western blot.

RESULTSDP-M&O decreased the levels of TNF-alpha, IL-1beta, IL-6, and IL-8 and increased that of IL-10 in a concentration-dependent manner. Significant differences were found between the 20% P-M&O and PAg groups in the release of the inflammatory cytokines TNF-alpha (70.8 +/- 22.3 vs. 277.1 +/- 65.5, P < 0.01) , IL-113 (277.5 +/- 22.6 vs. 630.4 +/- 89.7, P <0.01), IL-6 (232.7 +/- 62.7 vs. 994.2 vs. 182.3, P < 0.01), IL-8 (227.3 +/- 79.2 vs. 769.3 +/- 284.1, P < 0.01), and IL-10 (640.2 +/- 201.2 vs. 271.1 +/- 55.8, P < 0.01). Compared with the PAg group, the 10 and 20% P-M&O groups showed remarkable decreases in the protein expression of MCP-1/CCL2 in the RWPE-1 cells (1.12 +/- 0.34 vs. 0.56 +/- 0.11 and 0.34 +/- 0.08) and that of VCAM-1 in the EA. hy926 cells (0.94 +/- 0.22 vs. 0.52 +/- 0.17 and 0.38 +/- 0.12) (P < 0.05 or 0.01).

CONCLUSIONThe compatibility of musk and olibanum can decrease the expression of MCP-1/CCL2 in prostate cells and VCAM-1 in vascular endothelial cells, blocking the adhesion of leucocytes and suppressing inflammatory response.

Animals ; Blotting, Western ; Cytokines ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Fatty Acids, Monounsaturated ; pharmacology ; Frankincense ; pharmacology ; Inflammation ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; Male ; Mice ; Mice, Inbred BALB C ; Monocytes ; drug effects ; metabolism ; Prostate ; cytology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism