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.

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

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 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

OBJECTIVETo investigate if an adenovirus vector carrying antisense multidrug resistance gene 1 (MDR1) could reverse multidrug resistance (MDR) of HepG2/ adriamycin (ADM) cells in tumors transplanted in athymic mice.

METHODSAn adenovirus vector carrying AFP promoter and antisense MDR1 was constructed. HepG2 MDR cells (HepG2/ADM) were induced by graded resistance to ADM and were subcutaneously inoculated into athymic mice to construct the transplanted tumor. After adeno-asmdr1 was injected, the volume of the transplanted tumor and the apoptotic body in the xenograft tumor cells were observed and reverse transcriptase polymerase chain reaction was employed to investigate the expression of the mdr1-mRNA from the mouse transplanted tumor cells.

RESULTSFollowing injection with adeno-asmdr1, the tumor volumes in this mice group did not increase. However the tumor volume in the PBS plus ADM group did increase significantly (P less than 0.05). In the tumor xenograft cells, mdr1 mRNA in the xenografts was assessed by RT-PCR and found to be reduced at week 1, and at week 4 in the ADM+asmdr1 group, but it was stable in the ADM group. It was only 20% in the ADM+asmdr1 group compared to the ADM group at the 4th week. Evidence of apoptosis was observed in the tumor xenograft cells treated with adeno-asmdr1, but there was rarely any apoptosis in the group treated with ADM and PBS.

CONCLUSIONAdenovirus carrying antisense mdr1 RNA can partially reverse the MDR of HepG2/ADM cells and inhibit tumor growth by down-regulating mdr1 mRNA resulting in tumor cell apoptosis.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Adenoviridae ; genetics ; Animals ; Carcinoma, Hepatocellular ; drug therapy ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Genetic Vectors ; Hep G2 Cells ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; RNA, Antisense ; genetics

OBJECTIVEAdenovirus vector-mediated herpes simplex virus thymidine kinase gene (ADV-TK) transfer in combination with ganiciclovir (GCV) is one of the major gene therapy strategies to eradicate tumor cells. This study was aimed at determining the in vivo anti-tumor efficacy of ADV-TK in combination with ganiciclovir (GCV).

METHODSA murine xenotransplant model of human small-cell lung cancer was established. ADV-TK was administrated by intra-tumoral injection followed by intraperitoneal administration of GCV. The anti-tumor efficacy was evaluated using index of tumor volume, relative tumor volume, tumor weight, relative tumor proliferative rate, and tumor growth curve.

RESULTSIn the presence of GCV, ADV-TK effectively inhibited growth of human small-cell lung cancer in a dose-dependent fashion. An inhibition plateau was not observed within the current dosage range. ADV-TK at a dose of 6.0 x 10(9) viral particles/kg in the presence of GCV lead to 64.6% and 81.7% inhibition of tumor growth respectively in two independent experiments. ADV-TK or GCV alone caused slight inhibition of tumor growth, which was not statistically significant as compared to the negative control group (P > 0.05).

CONCLUSIONADV-TK followed by GCV is highly efficacious to inhibit the growth of human small-cell lung cancer in a murine xenotransplant model. The results presented here are encouraging to warrant a further clinical evaluation of the potential therapeutic benefits of this strategy.

Adenoviridae ; genetics ; Animals ; Carcinoma, Small Cell ; therapy ; Female ; Ganciclovir ; therapeutic use ; Genetic Therapy ; Humans ; Lung Neoplasms ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; Transplantation, Heterologous

OBJECTIVETo discuss the best surgical approach to the skull base neoplasms.

METHODSRetrospective analysis the 79 skull base neoplasms cases treated with surgical resection in Qilu hospital of Shandong university from 1992 to 2002. Eleven surgical approaches including midfacial degloving, frontal coronal discission, nasal eversion, maxillary swing, partial maxillary resection, total resection of orbit, mandibular swing, combination of front, temple, preauricular, post aureum, neck, and transoral approaches were used to resect the tumor which involved fossae pterygopalatine, paranasal sinuses, nasopharynx, antero, meso and posterobasilar region, lobi frontalis and lobi temporalis of cerebrum.

RESULTSSeventy-nine skull base neoplasms were totally removed and no one died from the operation. Although 5 cases complicated with cerebrospinal fluid leak and all recovered within 1 week, no serious cranium-cerebrum complication occurred. In 29 patients with benign tumor including 11 cases of meningioma, 3 cases of chondroma, 1 case of hemangio-meningioma, 1 case of cavernous hemangioma, 2 cases of osteodysplasia fibromas, 9 cases of neurofibroma, 1 case of glomus jugular tumor, 1 case of neurilemmoma, 19 have survived over 5 years and the longest one has survived over 8 years. For 50 patients with malignant tumor including 3 cases of well-differentiated squamous cell carcinoma, 17 cases of moderately differentiated squamous cell carcinoma, 11 cases of poorly differentiated squamous cell carcinoma, 1 case of undifferentiated carcinoma, 2 cases of chondrosarcoma, 5 cases of canceration of papilloma, 2 cases of adenocarcinoma, 1 case of esthesioneuroblastoma, 2 cases of malignant fibrohistiocytoma, 1 case of fibrosarcoma, 2 cases of malignant mixed tumour, 3 cases of sarcoma survival rates of 3 and 5 years were 59.2% (29/49), 38.5% (10/26) respectively.

CONCLUSIONIn order to resect the tumor completely and reduce the complication and malformation as far as possible, different surgical approaches must be designed according to the pathological changes characters and involved area,and the surgeon should select the shortest approach, avoid to damage the important neurovascular structure, and resect the tumor through the natural anatomy space by the shelter incision.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; methods ; Retrospective Studies ; Skull Base Neoplasms ; surgery