In the present study, the specifically knockdown models of P-gp or MRP2 were constructed by using a series of chemically synthesized small interfering RNA (siRNA) in vitro. The expression of P-gp and MRP2 was measured by real-time PCR and Western blot, and the function was evaluated by applying P-gp and MRP2 substrate, rhodamine and methotrexate. The results showed that MRP2 siRNA-3 or P-gp siRNA-2 significantly decreased the mRNA expression of MRP2 or P-gp, the inhibition ratio was 68% or 84%; MRP2 siRNA-3 or P-gp siRNA-2 at a dose of 80 nmol x L(-1) significantly reduced the protein expression of MRP2 or P-gp at 48 h after treatment, the inhibition ratio was 62% or 70%. Meanwhile, other transporters were not influenced by siRNA. When pretreatment with MRP2 siRNA-3 or P-gp siRNA-2, the efflux of methotrexate or rhodamine decreased significantly and the intra-cellular concentration increased. The results suggested that chemically synthesized siRNA could significantly inhibit the expression and function of MRP2 and P-gp, and the model of RNAi in vitro could be used to evaluate the role of efflux transporters in transportation of drugs.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Gene Knockdown Techniques ; Multidrug Resistance-Associated Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction

OBJECTIVETo compare the effect of laparoscopic radical operation and open operation on systemic immunity in patients with colorectal cancer.

METHODSSixty patients with colorectal cancer were randomly divided into laparoscopic and open operation groups from March 2004 to December 2004, each group had 30 cases. CRP, IgA, IgM, IgG, CD3 (+) cells, CD4 (+) cells, CD8 (+) cells, NK cells, CD4 (+) CD5RA (+) cells, CD4 (+) CD45RO (+) cells and lymphocytes in peripheral blood were counted and compared on the 1st day before operation, 3rd and 7th day after operation.

RESULTSThe two groups were comparable as for age, tumor location and stages. In open operation group, lymphocyte counts were (1.09+/- 0.29) x 10(9)/L, CD4 (+) cell (0.54 +/- 0.14) x 10(9)/L, CD8 (+) cell (0.31 +/- 0.08) x 10(9)/L, CD4 (+) CD45RO (+) (61.1+/- 8.9)%, and IgM level (136.9+/- 52.8) IU/ml, IgG (115.2 +/- 45.7) IU/ml on the 3rd day after operation, CD8 (+) cell counts were (0.32 +/- 0.09) x 10 (9)/L, CD4 (+) CD45RO (+) cell (63.2 +/- 9.1)% on the 7th day after operation, were all significantly lower than those on the 1st day before operation respectively(P< 0.05, P< 0.01). In laparoscopic operation group, the decreases of such parameters except CD4 (+) CD45RO (+) cell (62.7 +/- 12.5)% were not obvious on the 3rd day after operation. There were significant difference in lymphocyte counts (1.29 +/- 0.37) x 10( 9 )/L, IgM (164.5 +/- 48.2) IU/ml and CD8 (+) cell counts (0.38 +/- 0.09) x 10 (9) /L on the 3rd day after operation between two groups (P< 0.05).

CONCLUSIONCompared with open radical operation, laparoscopic radical operation has predominance in protecting systemic immunity to treat colorectal carcinoma.

Aged ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; immunology ; Colorectal Neoplasms ; immunology ; surgery ; Female ; Humans ; Killer Cells, Natural ; immunology ; Laparoscopy ; Lymphocyte Count ; Male ; Middle Aged

Acute Disease ; Biomarkers, Tumor ; analysis ; Child ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; Histocytochemistry ; Humans ; Leukemia ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis

OBJECTIVETo investigate the prevalence of aminoglycoside resistance and genotyping of acetyltransferase in Escherichia coli.

METHODSResistance phenotypes to 12 antibiotics of 44 Escherichia coli isolates were analyzed using agar dilution method and 3 aminoglycoside resistance genes aac(3)-I, II and aac(6')-I were determined by PCR method.

RESULTSIn 44 clinical isolates, the occurrence of ESBLs was 45.45%, resistance rates were discrepant for amikacin (18.18%), gentamicin (56.82%) and tobramycin (61.36%), the prevalence of phenotype TG (tobramycin and gentamicin) indicative of aac(3)-II production and TGA (tobramycin, gentamicin and amikacin) indicative of aac(6')-I production were 36.36% and 18.18%, respectively. The most common aminoglycoside resistance genotype of acetyltransferase was aac(3)-II (52.27%) and aac(6')-I was lower (29.55%), but no aac(3)-I was detected.

CONCLUSIONAt least 2 acetyltransferase genes exist in this area i.e. aac(3)-II and aac(6')-I.

Acyltransferases ; genetics ; Amikacin ; pharmacology ; Aminoglycosides ; pharmacology ; Drug Resistance, Bacterial ; genetics ; Escherichia coli ; enzymology ; genetics ; Genotype ; Gentamicins ; pharmacology ; Phenotype ; Tobramycin ; pharmacology

OBJECTIVETo objectively assess the clinical efficacy and safety of a new Pulian Ointment (, NPLO) in treating psoriasis of blood-heat syndrome of Chinese medicine.

METHODSA total of 108 patients with psoriasis of blood-heat syndrome were equally assigned, using a randomizing digital table, to the test group treated externally with NPLO and the control group treated with placebo; the medication was done using a singleblinded method twice a day. Meanwhile, all patients received by oral intake a conventional Chinese decoction for clearing heat and cooling blood; the therapeutic course was 4 weeks for both groups. The therapeutic efficacy, changes in the Psoriasis Area and Severity Index (PASI) score and various aspects of the lesion, including scaly eruption, erythema, infiltration, size, score of itching as well as adverse reactions were observed.

RESULTSThe trial was completed in 100 patients, 51 in the test group and 49 in the control group. The remarkably effective rate was 45.10% and the total effective rate was 84.31% in the test group, which were significantly higher than those in the control group, 12.24% and 51.02%, respectively, showing a significant difference between groups (P<0.01). The test group also showed better effects in the improvement of the PASI score of the lesions and scores on erythema, infiltration, size of lesion as well as itching. No adverse event was found in either group.

CONCLUSIONNPLO is a Chinese remedy for the external treatment of psoriasis of the blood-heat syndrome with a reliable therapeutic efficacy and good safety.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Ointments ; Psoriasis ; drug therapy ; Syndrome

Administration, Cutaneous ; Adult ; Analgesics, Opioid ; administration & dosage ; Blood Pressure ; drug effects ; Female ; Fentanyl ; administration & dosage ; Gynecologic Surgical Procedures ; Heart Rate ; drug effects ; Humans ; Middle Aged ; Pain, Postoperative ; drug therapy ; Respiration ; drug effects

Adolescent ; Adult ; Aged ; Ankle Joint ; pathology ; physiopathology ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Tuberculosis, Osteoarticular ; drug therapy ; pathology ; physiopathology ; surgery ; Young Adult

OBJECTIVETo develop a fast, high-throughput screening method with suspension array technique for simultaneous detection of biothreat bacteria.

METHODS16 S rDNA universal primers for Bacillus anthracis, Francisella tularensis, Yersinia pestis, Brucella spp.and Burkholderia pseudomallei were selected to amplify corresponding regions and the genus-specific or species-specific probes were designed. After amplification of chromosomal DNA by 16 S rDNA primers 341A and 519B, the PCR products were detected by suspension array technique. The sensitivity, specificity, reproducibility and detection power were also analyzed.

RESULTSAfter PCR amplification by 16 S rDNA primers and specific probe hybridization, the target microorganisms could be identified at genus level, cross reaction was recognized in the same genus. The detection sensitivity of the assay was 1.5 pg/microl (Burkholderia pseudomallei), 20 pg/microl (Brucella spp.), 7 pg/microl (Bacillus anthracis), 0.1 pg/microl (Francisella tularensis), and 1.1 pg/microl (Yersinia pestis), respectively. The coefficient of variation for 15 test of different probes was ranged from 5.18% to 17.88%, it showed good reproducibility. The assay could correctly identify Bacillus anthracis and Yersinia pestis strains in simulated white powder samples.

CONCLUSIONThe suspension array technique could be served as an opening screening method for biothreat bacteria rapid detection.

Bacillus anthracis ; isolation & purification ; Bioterrorism ; prevention & control ; DNA Primers ; DNA, Bacterial ; analysis ; Francisella tularensis ; isolation & purification ; Oligonucleotide Array Sequence Analysis ; methods ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; genetics ; Yersinia pestis ; isolation & purification

Cellular excitability is an important physiological factor in maintaining normal cardiac activity. The present study was designed to investigate the ionic mechanism underlying different excitability in atrial and ventricular myocytes of guinea pig heart using a whole-cell patch configuration. We found that excitability is lower in ventricular myocytes than that in atrial myocytes. Although the density of voltage-gated fast Na(+) current (INa) was lower in ventricular myocytes, it would not correlate to the lower excitability since its availability was greater than that in atrial myocytes around threshold potential. Classical inward rectifier K(+) current (IK1) was greater in ventricular myocytes than that in atrial myocytes, which might contribute in part to the lower excitability. In addition, the transient outward K(+) current with inward rectification (Itoir) elicited by depolarization was greater in ventricular myocytes than that in atrial myocytes and might contribute to the lower excitability. In ventricular myocytes, Ba(2+) at 5 µmol/L significantly inhibited Itoir, enhanced excitability, and shifted the threshold potential of INa activation to more negative, and the effect was independent of affecting INa. Our results demonstrate the novel information that in addition to classical IK1, Itoir plays a major role in determining the distinctive excitability in guinea pig atrial and ventricular myocytes and maintaining cardiac excitability. More effort is required to investigate whether increase of Itoir would be protective via reducing excitability.
Animals ; Atrial Function ; Guinea Pigs ; Heart Atria ; cytology ; Heart Ventricles ; cytology ; Myocytes, Cardiac ; physiology ; Potassium Channels, Inwardly Rectifying ; physiology ; Ventricular Function ; Voltage-Gated Sodium Channels ; physiology

For thousands of years, moxibustion has been used for various diseases in China and other Asian countries. Despite the recent surge in Chinese herbal studies, few randomized controlled trials have been conducted on this modality, possibly due to the lacking of suitable double blinding methodology. This is a review of extant sham moxa devices and an introduction to a recently developed device that needs further validation.
Double-Blind Method ; Humans ; Moxibustion ; Randomized Controlled Trials as Topic