OBJECTIVETo evaluate in vitro anti-tumor effect of chemotherapeutic drugs on human gastric cancer cells, and investigate the relationship with Bcl-2 expression.

METHODSSingle cell suspension was prepared from fresh gastric cancer tissue and exposed to taxol (Tax), 5-fluorouracil (5-FU), cisplatin (CDDP), adriamycin (ADM), mitomycin (MMC) respectively for 48 hours. Metabolic activity and inhibitory rate of cells were detected by MTT assay. Expression of Bcl-2 was examined with immunohistochemistry.

RESULTSThe inhibitory rates of cancer cells exposed to chemotherapeutic drugs were different and Tax, 5-FU, CDDP had remarkably higher rates than ADM and MMC. The lower differentiated gastric cancer cells were more sensitive than the higher ones. Positive expression rate of Bcl-2 was 80% and the positive cells showed resistance to 5-FU, ADM and MMC.

CONCLUSIONSChemosensitive testing by MTT assay can constitute the prediction for the application of chemotherapeutic drugs individually. Overexpression of Bcl-2 may contribute to multiple drug-resistance of tumors.

Adult ; Aged ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Cell Survival ; Cisplatin ; pharmacology ; therapeutic use ; Doxorubicin ; pharmacology ; therapeutic use ; Drug Screening Assays, Antitumor ; Female ; Fluorouracil ; pharmacology ; therapeutic use ; Humans ; Male ; Middle Aged ; Mitomycin ; pharmacology ; therapeutic use ; Mitomycins ; pharmacology ; therapeutic use ; Paclitaxel ; pharmacology ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; Tumor Cells, Cultured

OBJECTIVETo investigate the reversing effects of curcumin on hepatocellular carcinoma drug resistance Bel7402/5-Fu cell line.

METHODSThrough the exposure to gradual increased concentrations of 5-fluorouracil (5-Fu), the cell line Bel7402 was induced to establish a multi-drug resistant sub-cell line Bel7402/5-Fu. The sensitivity to 6 chemotherapeutics of Bel7402 and Bel7402/5-Fu were detected using methyl thiazolyl tetrazolium (MTT) assay. The 50% inhibitory concentration (IC50) and resistant index (RI) were calculated. The differences of the inhibition ratio of Bel7402/5-Fu by curcumin, 5-Fu, curcumin combined with 5-Fu were detected using MTT assay. The effects of curcumin, 5-Fu, curcumin combined with 5-Fu on the Bel7402/5-Fu apoptosis were detected using flow cytometry.

RESULTSThe Bel7402/5-Fu cell line showed multi-drug resistance (MDR) to various chemotherapeutics, with the highest RI shown of 5-Fu (being 109.55 +/- 14.30 times). The inhibition ratio of 5, 10, and 20 microg/mL curcumin combined with 5-Fu (50% IC50) was respectively 21.47% +/- 1.49%, 27.10% +/- 2.32%, and 59.37% +/- 2.45%. The Bel7402/5-Fu apoptosis ratio of 5, 10, and 20 microg/mL curcumin combined with 5-Fu (50% IC50) was 30.92% +/- 2.10%, 44.87% +/- 2.24%, and 50.36% +/- 2.58%, respectively, which was obviously higher than that of the curcumin group and the 5-Fu group. Besides, the apoptosis rate increased along with increased curcumin concentration in the range of 0 -20 microg/mL.

CONCLUSIONCurcumin could induce the apoptosis of Bel7402/5-Fu. Meanwhile, it showed favorable reversing effects on MDR.

Cell Line, Tumor ; Curcumin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Fluorouracil ; pharmacology ; Humans

OBJECTIVETo study the role of cerebrovascular hemodynamic indexes (CVHI) changing in stroke and to provide reference for stroke prevention and risk factor study.

METHODSFrom 2003 to 2004, participants aged 40 years and above in two communities in Fengxian district were recruited by cluster sampling. Risk factors of stroke and CVHI were investigated and checked during baseline investigation. A total of 10 565 individuals completed the survey and met the inclusion criterion. After baseline investigation, the cohort was followed up for stroke occurrence. Relative risk (RR) of CVHI and common risk factors were estimated by cohort study design.

RESULTSAge of the cohort was (56.2 ± 11.4) years. 4444 (42.1%) were males and 6121 (57.9%) were females. Total follow-up duration was 67 885.7 person-years. A total of 195 stroke cases occurred and incidence density of stroke was 287.2 per 100 000 person-years. Stroke incidence in exposure groups of hypertension, heart disease and alcohol drinking was 3.47% (108/3118), 2.96% (21/710) and 2.50% (47/1882), respectively. The incidence in corresponding non-exposure group was 1.17% (87/7448), 1.77% (174/9855) and 1.70% (148/8683) respectively. There was significant difference between 2 groups (χ(2) value was 62.72, 4.56 and 4.94, respectively, P < 0.05). Stroke incidence in CVHI score < 25, 25 - 49, 50 - 74 and ≥ 75 groups was 9.12% (59/647), 5.68% (44/775), 2.52% (39/1545) and 0.72% (53/7403)(χ(2)trend = 273.57, P < 0.05), respectively. Incidence of stroke in 40 - 49, 50 - 59, 60 - 69, ≥ 70 years age group was 0.22% (8/3565), 1.28% (43/3357), 2.71% (50/1848) and 5.88% (94/1600) (χ(2)trend = 181.48, P < 0.05), respectively. Multiple Cox regression analysis indicated that RR (95%CI) value of hypertension and cigarette smoking was 1.40(1.02 - 1.92) and 1.59(1.19 - 2.12), respectively when comparing with non-exposure group. RR (95%CI) value in CVHI score < 25, 25 - 49 and 50 - 74 points group were 6.15 (4.08 - 9.26), 4.55 (2.98 - 6.96) and 2.68 (1.75 - 4.09), respectively when comparing with the score ≥ 75 points group. RR (95%CI) value in age 50 - 59, 60 - 69 and ≥ 70 years group was 4.61 (2.16 - 9.82), 7.81 (3.67 - 16.60) and 13.49(6.44 - 28.24), respectively when comparing with below 40 years group.

CONCLUSIONCVHI score is the strong independent predictive factor and hypertension, cigarette smoking and age are the independent risk factors of stroke.

Aged ; Brain ; physiopathology ; Cohort Studies ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Risk Factors ; Stroke ; epidemiology ; etiology ; physiopathology

OBJECTIVEIn this study we investigated the functional restoration of nonsense mutations in the SCN5A gene.

METHODSThe readthrough-enhancing reagents were introduced to HEK293 cells to suppress one nonsense mutation W822X in the SCN5A gene. Patch-clamp was used to record the whole-cell current and dynamics. Western blot and immunofluorescence staining were used to certify the expression and the location of the sodium channel.

RESULTSIn transfected HEK293 cells, the nonsense mutation in SCN5A inhibited the expression level of full-length protein, and the sodium currents from the mutant channels were less than 3% of the wild-type level. Readthrough enhancement by decreasing translation termination efficiency with a siRNA targeting eukaryotic release factor eRF3a (a GTPase that binds eRF1), the sodium current from the mutant cDNAs was restored to as much as 30% of the wild-type. After the treatment by the readthrough-enhancing reagents, the channels from cDNA carrying W822X remained the features of wild-type phenotype, and Western blot and immunochemical staining also showed the expression of full-length channel proteins.

CONCLUSIONReadthrough-enhancing reagents could effectively suppress nonsense mutations in SCN5A and partially restore the function of sodium channel and the expression of full-length channels.

Codon, Nonsense ; HEK293 Cells ; Humans ; NAV1.5 Voltage-Gated Sodium Channel ; Patch-Clamp Techniques ; Plasmids ; RNA, Small Interfering ; Sodium Channels ; genetics ; metabolism ; Transfection

OBJECTIVETo search for a non-medicine therapy for infant emaciation.

METHODSThe observation group of 55 cases were treated with auricular point sticking combined with chiropractics. For the auricular point sticking therapy, vaccaria seeds were stuck at 9 points such as small intestine, Shenmen, spleen, endocrine, and others, and replaced once every 5 days, 10 days constituting one course. Chiropractics: knead or massage the muscles from lumbosacral area upward to Dazhui (GV 14). The control group were treated with oral administration of pepsin mixture, 10 mL after meals, thrice each day. They were treated for 5 courses.

RESULTSOf the 55 cases in the observation group, 7 were cured, 20 markedly effective, 24 effective and 4 ineffective, with a total effective rate of 92.7%, and among the 23 cases in the control group, 2 cases were cured, 7 markedly effective, 8 effective and 6 ineffective, with a total effective rate of 73.9%.

CONCLUSIONAuricular point sticking combined with chiropractics is a better non-medicine therapy for emaciation in children.

Acupuncture, Ear ; Child ; Child, Preschool ; Combined Modality Therapy ; Emaciation ; therapy ; Female ; Humans ; Male ; Manipulation, Chiropractic

JAK-3, a member of the Janus kinase family, is a protein tyrosine kinase, which plays an important role in the JAK-STAT signaling pathway. Previous studies showed that regulation of JAK-3's activity plays a crucial role in the treatment of diseases such as rheumatoid arthritis. Many reports have been published with a focus on selective JAK-3 inhibitors, some of which showed excellent JAK-3 selectivity and inhibitory activities. Among the JAK-3 inhibitors reported, tofacitinib has satisfactory therapeutic benefits in the clinical trials, and has been approved for treatment of patients with rheumatoid arthritis. However, some JAK-3 inhibitors exhibited moderate to severe side effects, which need to be controlled by drug improvement. In order to pave the way for improvement of current JAK-3 inhibitors and development of new JAK-3 inhibitors, we provide an outline of the structure of JAK-3 and strategies in development of its inhibitors.

OBJECTIVETo evaluate in vitro antitumor effects of chemotherapeutic drugs, and investgate the relationship with expression of hTERT mRNA in human gastric cancer tissues.

METHODSFresh samples of gastric cancer obtained from operation room were prepared to single-cell suspension (3 x 10(5) to 5 x 10(5) cells ml(-1)) and were separately exposed to taxol (TAX), cisplatin (CDDP), 5-fluorouracil (5-Fu), adriamycin (ADM), mitomycin (MMC) for 48 hours. Metabolic activity and inhibitory rate of the cells were determined by trypan blue exclusion and MTT assay. Expression of hTERT mRNA was detected by in situ hybridization (ISH).

RESULTSThe inhibition rate of cancer cells exposed to chemotherapeutic drugs was different, and that of TAX, CDDP, 5-Fu was significantly higher than that of ADM and MMC. The positive rate of hTERT mRNA expression was 90.0% (54/60) and positive cells showed resistance to 5-Fu and ADM.

CONCLUSIONOverexpression of hTERT mRNA may contribute to primary drug-resistance of tumors. Chemosensitivity tests by MTT assay may contribute to prediction of effectness in applying chemotherapeutic drugs and identify drug-resistant cases.

Adenocarcinoma, Mucinous ; metabolism ; pathology ; Adenocarcinoma, Papillary ; metabolism ; pathology ; Adult ; Aged ; Antibiotics, Antineoplastic ; pharmacology ; Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Carcinoma, Signet Ring Cell ; metabolism ; pathology ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Female ; Fluorouracil ; pharmacology ; Humans ; Male ; Middle Aged ; Mitomycin ; pharmacology ; Paclitaxel ; pharmacology ; RNA, Messenger ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Telomerase ; genetics ; metabolism

OBJECTIVETo investigate the clinical usefulness of noninvasive diagnostic methods in evaluating liver fibrosis in hepatitis B virus (HBV) patients.

METHODS102 patients with chronic hepatitis B (CHB) were enrolled from Beijing Friendship Hospital Affiliated to Capital University of Medical Sciences. Noninvasive diagnostic methods including ultrasonography, CT, serum markers of liver function and fibrosis, and HBV DNA were performed and compared with histological fibrotic changes in order to establish a noninvasive method for detecting the degree of liver fibrosis.

RESULTSThe total score of liver surface, edge, parenchyma echogenicity, intrahepatic vessels, and the size of spleen had a coefficient of 0.822 with fibrotic stage. By receiver operating curve (ROC) analysis, the sensitivity to distinguish cirrhosis from CHB was 86.1% and the specificity was 95.5% if the total ultrasonic score was more than 10. The CT imaging diagnosed liver cirrhosis with a specificity of 100% and a sensitivity of 48.5%. The change of CT values in cirrhotic patients was lower than that in controls and no cirrhotic patients (F=5.805, P<0.01), when the voltage was increased from 100 KV to 140 KV. Except normal controls and S1 group, S2 and S3 group, the level of HA and collagen IV between the other groups were statistically different. The cut-off value of HA to diagnose cirrhosis was 108 (microg/L) with a sensitivity of 72.2% and a specificity of 80.3%. The cut-off value of collagen IV to diagnose cirrhosis was 188 (microg/L) with a sensitivity of 72.2% and a specificity of 78.8%. When ultrasonography was combined with serum markers, the sensitivity was 72.2% and the specificity was 80.3%.

CONCLUSIONBoth ultrasonography and serum markers are useful to diagnose cirrhosis. The combination of the two examinations is more valuable than any one alone. The characteristic CT imaging has high specificity but low sensitivity in diagnosing early cirrhosis. HA and collagen IV are correlated more closely with the stage of fibrosis, and can reflect the severity of fibrosis.

Adolescent ; Adult ; Aged ; Biomarkers ; blood ; Collagen Type IV ; blood ; Female ; Hepatitis B, Chronic ; diagnostic imaging ; pathology ; Humans ; Hyaluronic Acid ; blood ; Liver Cirrhosis ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Sensitivity and Specificity ; Ultrasonography

OBJECTIVETo investigate the effect of gene Af116609 on gastric cancer multi-drug resistance (MDR) by introducing it into gastric cancer multi-drug resistant (MDR) cell line SGC7901/VCR.

METHODSGene Af116609 was cloned from SGC7901/VCR by RT-PCR and its differential expression between gastric cancer MDR cells and its parental cells was displayed by Northern blot. The gene was introduced to gastric cancer cells by transfection of recombinant eukaryotic expression vector by electroporation. MTT assay in vitro was applied to investigate its effect on multi-drug resistance phenotype of gastric cancer cells.

RESULTSThe full length CDS of gene Af116609, as long as 327 bp, was cloned from gastric cancer MDR cell line SGC7901/VCR and its sequence was coincident with the hypothetical gene Af116609 in GenBank. It was overexpressed in MDR cells than its parental cells at mRNA level. In the MTT assay in vitro, the drug sensitive cells transfected with sense eukaryotic expression vector showed upregulated targeted gene, with increased resistance to vincristine, 5-fliorouracil and arabinoside, and decreased resistance to adriamycin, but no influence on resistance to methotrexate. However, the drug resistant cells transfected with anti-sense eukaryotic expression vector, showed down regulated targeted gene, with less resistance to all the five anticancer drugs to different degrees.

CONCLUSIONGene Af116609 is related to MDR phenotype of gastric cancer cells and may become a candidate molecular target to reverse the MDR of gastric cancer.

Antineoplastic Agents, Phytogenic ; pharmacology ; Autoantigens ; genetics ; Cell Line, Tumor ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Humans ; RNA, Small Cytoplasmic ; genetics ; Ribonucleoproteins ; genetics ; Stomach Neoplasms ; genetics ; pathology ; Vascular Endothelial Growth Factor A ; biosynthesis ; Vincristine ; pharmacology

Adult ; Aged ; Aneurysm ; diagnosis ; pathology ; Computed Tomography Angiography ; methods ; Female ; Humans ; Male ; Middle Aged ; Splenic Artery ; pathology