Objective To establish the method of fingerprint analysis for volatile oil from Cinnamomum cassia Presl and to determine the main characteristic components.Methods The main components of the volatile oil of Cinnamomum cassia Presl from different habitats of Guangdong and Guangxi provinces and from GAP base were compared by GC fingerprinting,and 11 common components were determined.The chromatogram conditions were as follows: the GC system consisted of Flame Ionization Detector(FID) and HP6890 gas chromatograph with a HP-5 column(Crosslinked Mehyl siloxame,30 m? 0.321 mm? 0.25 ? m),the temperatures of sample vent and FID were 240 ℃ and 300 ℃ respectively and the column programmed temperature was elevated from 100 ℃ to 140 ℃ at the rate of 20 ℃ ? min-1 and then from 140 ℃ to 200 ℃ at the rate of 2 ℃ ? min-1,the carrier gas was N2 and its flow rate was 0.4 mL? min-1,and the split ratio was 50 ∶ 1.Results With 11 components as indexes,the RSD of precision,reproducibility and stability of GC fingerprinting method is in the range of 5 %.Conclusion A good fingerprint of Cinnamomum cassia Presl has been established.The method is reliable,accurate and can be applied for the quality control of Cinnamomum cassia Presl.

This study was aimed to establish a high performance liquid chromatography (HPLC) method coupled with Evaporative Light-scattering Detector (ELSD) in order to develop the determination of fingerprint of terpene lactones in Ginkgo biloba tablets. An Agilent Extend-C18 (4.6 mm í 250 mm, 5 μm) was employed as the analysis column and the normal propyl alcohol-tetrahydrofuran-water (1:15:84) as mobile phase. The column temperature was 30℃. And the flow rate was 1.0 mL·min-1. HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS) was used to identify the common peaks. The fingerprint was further evaluated by chemometrics methods including principal component analysis (PCA), similarity analysis (SA) and hierarchical clus-tering analysis (HCA). The results showed that the precision, stability and repeatability of this method were favorable. Five common peaks were identified by LC/Q-TOF MS as ginkgolide J ( M ) , C , A , B and bilobalide , respectively . Fourteen batches of Ginkgo biloba tablets were determined. With the aid of PCA, SA and HCA, the common pattern of the fingerprint of terpene lactones was established. Samples were divided into 4 clusters by their quality differ-ence. It was concluded that the method established in this paper can be used for quality evaluation of terpene lac-tones in G ink go b ilob a tablets.

The effects of Guizhi Fuling capsule and its active ingredient combination within different concentration on SPL proliferate were observed by MTT method. The ratio of CD80/86, CD3CD25 and CD3CD69 was used to evaluate cell activation effects of Guizhi Fuling capsule and its active ingredient combination by FCM. Guizhi Fuling capsule with concentration of 400 mg · L(-1)can promote spleen lymphocyte proliferation, as well as the active ingredient combination, which showed the obvious dose-effect relationship. Compared with control group, the difference has statistical significance (P≤0.01). The result of FCM showed that Guizhi Fuling capsule and its active ingredient combination can promote CD80 and CD86 expression on spleen lymphocyte, and also can increase CD25 and CD69 ratio between spleen CD3+ cells. Compared with control group, the difference has statistical significance (P≤0.01). Thus, Guizhi Fuling capsule and its active ingredient combination may have immune-modulate effects, and the mechanism may have a close relationship with the lymphocyte activation.
Animals ; Capsules ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Immunologic Factors ; pharmacology ; Lymphocyte Activation ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; drug effects ; immunology

Opioids are the main treatment drugs for the patients with moderate and severe cancer pain,and methadone is one of opioid drugs. Compared with the other opioids, methadone has the advantages of low cost, high oral bioavailability and better effect on neuropathic pain. However, methadone also shows such disadvantages as prolonged QT interval and large individual variance in pharmacokinetics. Although the safety of methadone has no difference from the other opioids in the treatment of cancer pain,the lack of knowledge and experience still limits its clinical application. This article aimed to summarize the clinical application of methadone in cancer patients,so as to help clinicians understand and apply methadone better.

OBJECTIVEAcute intracranial hypertension/cerebral edema (ICH/CE) is an increase in brain volume caused by an absolute increase in cerebral tissue water content. Severe ICH/CE is often associated with a higher mortality and higher neurological consequence rate in intensive care unit. However, little relevant information is available on critical condition of central nervous system in children. The aim of this survey was to study the causes, clinical epidemiology and risk factors of critical illness with ICH/CE in pediatric intensive care unit (PICU).

METHODSCase records of critically ill patients with ICH/CE admitted to PICU in Children's Hospital Affiliated to Shanghai Jiaotong University during the period from January, 1999 to December, 2003 were reviewed for causes, case fatality rate, prognosis and relationship with multiple organ dysfunction syndrome (MODS). Univariate analyses were performed to identify risk factors associated with ICH/CE.

RESULTSDuring the 5 years, 1446 cases with critical illnesses were admitted and ICH/CE developed in 216 patients. The leading causes of ICH/CE were central nervous system infection (27.8%), accidental injuries (22.4%), and sepsis (10.2%). The overall mortality of the patients with ICH/CE was 29.2%. The mortality showed no significant change during the years from 1999 to 2003 (chi(2) = 0.371, P = 0.985). There was no significant difference in mortality of patients with ICH/CE between those with and without neurological diseases (chi(2) = 0.546, P = 0.460). Univariate analyses involving 12 factors indicated the following risk factors: younger age, number of failed organ, lower pediatric critical illness score, underlying diseases, abnormal respiration and change in size of pupil (P < 0.05 or < 0.001). The following factors were not associated with higher risk of death from ICH/CE: sex, organ of primary disease, Glasgow coma score ( 7) on admission, elevated blood pressure and anterior fontanelle change (P > 0.05).

CONCLUSIONSThe mortality of ICH/CE remains high since 1999. Central nervous system infection, accidental injuries, and sepsis were leading causes of ICH/CE in PICU of the hospital. Children who had ICH/CE due to younger age, lower pediatric critical illness score, and complicated with MODS had a higher mortality rate.

Acute Disease ; Brain Edema ; mortality ; Child ; China ; epidemiology ; Critical Illness ; Hospitals, University ; Humans ; Intensive Care Units, Pediatric ; statistics & numerical data ; Intracranial Hypertension ; mortality ; Prognosis ; Retrospective Studies ; Risk Factors

OBJECTIVETo investigate the therapeutic efficacy of standard antiviral therapy applied after interferon (IFN) treatment failure in patients with chronic hepatitis C (CHC).

METHODSCHC patients who completed a 48-week course of IFN therapy (pegylated (Peg)-IFNa-2a at 180 mug, qw, ih with or without ribavirin (RBV) at 15 mg/kg/w) in our hospital between January 2009 and June 2012 but who showed no response (at week 48) or who relapsed (at week 72) were enrolled in the study. Prior to initiating the 48-week course of retreatment therapy (Peg-IFNa-2a plus RBV as above), the hepatitis C virus (HCV) genotype was detected and the viral load measured (baseline) by PCR of HCV RNA. Each patient's response to therapy was classified as follows: baseline vs. week 4 (rapid virological response, RVR), vs. weeks 12 and 24 (early virological response, EVR), vs. week 48 (end of treatment virological response, ETVR) and vs. week 72 (sustained virological response, SVR).

RESULTSOf the total 235 cases administered retreatment therapy, 60.0% (n = 140) achieved RVR, 77.4% (n = 182) achieved EVR, 83.8% (n = 197) achieved ETVR, 68.0% (n = 68%) achieved SVR, and 15.7% (n = 37) relapsed. Stratification analysis of recurrence (n = 158) and non-responsive (n = 77) sub-groups showed that the recurrence group experienced significantly higher rates of RVR, EVR, ETVR and SVR, but a significantly lower rate of relapse. Stratification analysis of genotype 1b carrier (n = 206) and non-1b carrier (n = 29) sub-groups showed that the 1b carriers had significantly lower rates of RVR, EVR, ETVR and SVR, but a significantly higher rate of relapse. Finally, the patients who achieved RVR (vs. non RVR, n = 95) and EVR (vs. non-EVR, n = 53) showed higher rates of SVR and ETVR.

CONCLUSIONCHC patients who fail to respond to the initial course of standard IFN-based therapy may achieve SVR upon retreatment, especially those infected with the HCV genotype 1b.

Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Genotype ; Hepacivirus ; genetics ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferon-alpha ; administration & dosage ; therapeutic use ; Interferons ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; administration & dosage ; therapeutic use ; Recombinant Proteins ; administration & dosage ; therapeutic use ; Retreatment ; Ribavirin ; administration & dosage ; therapeutic use ; Treatment Failure

The aim of the present study was to gain an insight into the thiopurine methytransferase (TPMT) genotyping assay, which was based on polymerase chain reaction (PCR), allele-specific PCR, restriction digestion of PCR products, denaturing high-performance liquid chromatography (DHPLC) and SNaPshot sequencing and in combination with direct DNA sequencing. Among the f our methods to test TPMT genetic SNPs based on PCR, allele specific PCR was not able to differentiate wild type from varied type. BsiYI, MwoI and AccI to digest PCR products were used so that SNP in TPMT exon 5, 7 and 10 tested. It showed that there were no differences between the results of digestion of PCR products and those of DNA sequence analysis. Therefore, this method was reliable. But some other methods were still needed to look for a compensation, because no restriction map changing resulted from the 2 SNPs in TPMT promotor was found. As to the results of DHPLC, those for the screening of TPMT exon-5 and -10 for SNPs were the same as restriction analysis of PCR products and direct DNA sequencing. But the variation of the heterozygotes in exon-7 was high, which was different from the results of direct DNA sequencing. After changing the Tm of DNA step by step, It was found that all the samples showed single peak when the temperature was 54 degrees C. But this result was unbelievable because a heterozygote in exon 7 as positive control could not be found. Therefore, it was necessary to test the sensitivity and accuracy of DHPLC, though DHPLC could be used as an effective method of SNPs screening. The results of the SNaPshot sequencing were also same as those of restriction analysis of PCR products and direct DNA sequencing. And the results showed that the bases of TPMT promoter -91 and -168 were G, instead of A and T. The results of the four methods to detect TPMT genetic SNPs based on PCR showed that SNPs analysis technique should be a combination of the techniques above-mentioned. One technique alone could not satisfy the need in clinics and research. The compensation of each other was very important.
Chromatography, High Pressure Liquid ; Exons ; Humans ; Methyltransferases ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide

OBJECTIVETo investigate the renal protective activity of Hsian-tsao Mesona procumbens Hemsl. water extracts in diabetic rats.

METHODSThirty Sprague-dawley female rats were randomly divided into three groups (n = 10 each), "control group" with intraperitoneal saline injection, "diabetic group" with 60 mg of intraperitoneal streptozotocin injection per kg of body weight and "Hsian-tsao group" with intragastric administration of Hsian-tsao extraction everyday for 4 weeks after intraperitoneal streptozotocin injection. The body weight and blood sugar were measured before and after model induction in the three groups. Thrombospondin-1 (TSP-1) expressions in the kidney were monitored by immunohistochemistry. Kidney ultrastructural changes were also analyzed by using transmission electron microscopy.

RESULTSBefore diabetic model induction, there were no significant differences among the three groups in body weight and blood sugar. Four weeks after the induction of diabetes, the differences became statistically significant. Electron microscopy also revealed disruption of the foot processes of the podocytes and other damages in diabetic group. These damages were significantly less severe in Hsian-tsao group when compared with the diabetic group. TSP-1 expressions in the kidney were significantly increased in both the diabetic group and Hsian-tsao group, but it was relatively lower in Hsian-tsao group than in diabetic group.

CONCLUSIONOur results showed that Hsian-tsao treatment in the diabetic rats effectively prevented the pathological alterations in the kidney and decreased the TSP-1 expression. It was suggested that Hsian-tsao had protective effect on the kidneys of the diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; pathology ; Diabetic Nephropathies ; metabolism ; pathology ; prevention & control ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Kidney ; drug effects ; metabolism ; pathology ; ultrastructure ; Lamiaceae ; chemistry ; Rats ; Rats, Sprague-Dawley ; Thrombospondin 1 ; metabolism

OBJECTIVETo study the changes of cardiac function following treatment with granulocyte colony stimulating factor (G-CSF) in patients with heart failure after myocardial infarction.

METHODSThirty-eight patients with heart failure after myocardial infarction were randomized into G-CSF treatment group and control group. All the patients received conventional treatment (medication and interventional therapy), and the patients in treatment group were given additional G-CSF (600 µg/day) for 7 consecutive days. The plasma level of brain-type natriuretic peptide (BNP) and the number of endothelial progenitor cells (EPCs) in the peripheral blood were detected before and at 7 days and 4 months after the treatment. The cardiac functions (LVSD, EDV, and LVEF) were evaluated by ultrasonic imaging before and at 2 weeks and 4 months after the treatment.

RESULTSThe number of EPCs was significantly higher in the treatment group than in the control group after the treatment especially at 7 days (P<0.01). In both groups, BNP level was lowered significantly after the treatment to recover the normal level (P<0.01). The cardiac functions were improved in all the patients at 7 days and 4 months after the treatment, and the improvement was more obvious in the treatment group (P<0.05), especially in terms of LVEF at 4 months after the treatment (P<0.01).

CONCLUSIONEPC mobilization by G-CSF can effectively improve the cardiac functions and lessen ventricular remodeling in patients with heart failure after myocardial infarction.

Aged ; Endothelial Cells ; cytology ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Heart Failure ; etiology ; physiopathology ; therapy ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; Myeloid Progenitor Cells ; cytology ; Myocardial Infarction ; complications ; physiopathology ; therapy ; Natriuretic Peptide, Brain ; metabolism ; Treatment Outcome ; Ventricular Remodeling

OBJECTIVETo investigate the effect of gastrin on invasiveness of human colon cancer cells and the role of gastrin receptor-focal adhesion kinase (FAK) signal transduction pathway in this proess.

METHODSpCR3.1/GR vector expressing gastrin receptor was transfected into a colorectal cancer cell line Colo320 with lipofectamine 2000, and screened by G418. The expression levels of gastrin receptor of the parental cell line Colo320 and the transfected cell line Colo320/GR were assayed by RT-PCR. On the other hand, antisense oligonucleotide of FAK was used to block its expression. The mock transfected Colo320 and sense oligonucleotide Colo320 cells were used as controls. Colo320 and Colo320/GR cells were treated with increasing doses (0 approximately 100 nmol/L) of gastrin. Invasiveness of Colo320 and Colo320/GR cells was determined by Boyden chamber. Phosphorylation of focal adhesion kinase (FAK) tyr-397 was examined by immunoprecipitation and Western-blot.

RESULTSRT-PCR results showed that the Colo320/GR cells had an mRNA level four times as high as that of Colo320 cells. Western blot showed that FAK tyr397 phosphorylation of Colo320 cells was apparently decreased. Colo320 and Colo320/GR cells showed a dose-dependent response to gastrin on invasiveness and phosphorylation of FAK tyr-397. Invasiveness of Colo320 cells reached its climax when concentration of gastrin was 100 nmol/L, and FAK tyr-397 phosphorylation was marked when concentration of gastrin was 10 nmol/L, but the latter decreased when gastrin concentration was increased to 100 nmol/L. Colo320/GR cells had the same tendency as Colo320 cells, but showed an even stronger invasiveness and a higher level of FAK tyr-397 phosphorylation than Colo320 cells. Before gastrin stimulation, the invasiveness of Colo320 cells transfected with antisense oligonucleotides and the controls showed no difference. After gastrin stimulation, the increase in invasiveness was much less than that in the controls.

CONCLUSIONGastrin can evidently promote invasiveness of Colo320 cells via gastrin-gastrin receptor-FAK signal transduction pathway.

Cell Line, Tumor ; Colorectal Neoplasms ; pathology ; Focal Adhesion Protein-Tyrosine Kinases ; metabolism ; Gastrins ; pharmacology ; Humans ; Neoplasm Invasiveness ; Signal Transduction