Tissue blotting, based on Enzyme-Linked Immunosorbent Assay (ELISA), is a technique for detection of plant viruses. This technique is not only high sensitivity and specificity, but also simpler and more rapid for detection. Samples that are blotted on membrane can be kept over three months. The results can directly display the section of virus infected. It is especially suitable for detection of plant viruses on a large scale.

Adult ; Arsenic Poisoning ; pathology ; Chronic Disease ; Hemosiderin ; metabolism ; Hemosiderosis ; metabolism ; pathology ; Humans ; Hypertension, Portal ; chemically induced ; metabolism ; pathology ; Liver Cirrhosis ; chemically induced ; metabolism ; pathology ; Male ; Pancytopenia ; chemically induced ; metabolism ; pathology ; Splenomegaly ; chemically induced ; metabolism ; pathology

Objective To investigate the suppression effect of expressing parvovirus H-1 nonstructural protein 1 (NS1) gene on human gastric cancer cells and the possible mechanisms.Methods A recombinant enhanced green fluorescent protein (eGFP) labeled NS1 of parvovirus H-1 plasmid was constructed.Human gastric cancer cell line SGC7901 was transfected with recombinant plasmid (experiment group) or blank vector (negative control group) and blank control group was treated with equal amount of phosphate buffered saline (blank control group).After transfection,the distribution of fluorescent signal was observed under fluorescent microscope.The expression of NS1 at gene and protein level was measured.Cell growth curve of each group was drawn.The expression of cell senescence-associated β-galactosidase (SA-β-Gal) was tested.The changes of cell cycle were investigated by flowcytometry.Two groups' comparision was performed by t-test.Results After transfection,NS1 was expressed in SGC7901 cells at gene and protein level.Compared with negative control group,the fluorescent signal accumulated in cell nucleus in experiment group.The percentage of SA-β-Gal positive cell in experiment group ((30.5 ± 1.4) ％) was higher than that of negative control group ((4.4± 1.1) ％) and the difference was statistically significant (t =-12.931,P ＜ 0.01).The growth inhibition rate of SGC7901 cells from the first day to the fourth day was 45％,62％,73％ and 77％,respectively.The cell cycle of eGFP-NS1 expressed SGC7901 cells was arrested at G0/G1 phase.Conclusion Parvovirus H-1 NS1 play the role in cell nucleus of gastric cancer cell line SGC7901 and could make cell cycle arrested at G0/G1 phase,which effectively inhibited the proliferation SGC7901 cell.

Aged ; Humans ; Male ; Pharyngeal Neoplasms ; Polyps ; Pyriform Sinus ; pathology

Objective:To observe the effects of soybean isoflavones(SI) on uteri and uterine Er?,ER? expression in perimenopausal rats. Method:Female SD rats in age of 7-month-old were used as control and 11-month-old SD rats were divided into 5 groups,model group,diethylstilbestrol group and 3 test groups exposed to three doses of SI(ig:30,12,4.6 mg/kg?d) respectively for 35 d. Serum concentrations of estradiol(E2) ,testosterone(T) ,follicle-stimulating hormone(FSH) and luteinizing hormone(LH) were determined. Ultrastructure of uterine luminal surface was observed by scanning electron microscopy and hematoxylin-eosin staining. Expression of ER? and ER? in the uteri were determined by immunohistochemistry. Results:Low dose SI significantly increased serum E2(P

OBJECTIVETo develop an HILIC method for determination of dencichine in Sanqi tablet and evaluate the quality of Sanqi tablet of different hatches from various manufactures in the market.

METHODThe chromatographic separation was conducted on a Thermo HILIC column (4.6 mm x 250 mm, 5 microm) kept at 25 degrees C with acetonitrile and 0.1% H3PO4 (60:40) as the mobile phase. The flow rate was set at 1 mL x min(-1) and the detection wavelength was set at 213 nm.

RESULTThe contents of dencichine in Sanqi tablet ranged from 1.60 to 4.31 mg x g(-1).

CONCLUSIONThe well established method was successfully applied to determine dencichine in Sanqi tablet. The results demonstrated that this method was simple, accurate and could be applied for quality control of Sanqi as well as its associated preparations.

To study the relationship between changes of microbial ATP in four kinds of murine tissues and the postmortem interval (PMI), healthy SD rats were sacrificed and their muscles, livers, spleens and kidneys were sampled at different postmortem intervals. The concentration of microbial ATP was detected using bioluminescent assay and the data was statistically analyzed. The concentration of microbial ATP in muscle increased with PMI time. The peak appeared at the 7th day after death, and at the 10th day, microbial ATP in muscle tissue increased again. In internal organs, the peaks of microbial ATP were observed at the 8th day after death and the level decreased during 8-10 d. The differences in microbial ATP concentration in liver, spleen and kidney were not statistically significant. During day 0 to day 9 after death, the correlation was best between PMI and microbial ATP in muscle. With PMI as the independent variable, the cubic polynomial regression equation was Y=0.02X(3)-0.166X(2)-0.666X+13.412 (R (2)=0.989, P<0.01). In internal organs, the best correlation was found between PMI and microbial ATP during day 0 to day 10. With PMI as the independent variable, the cubic polynomial regression equation was Y=0.016X(3)-0.127X(2)-0.809X+13.324 (R (2)=0.986, P<0.01). There existed high correlations between PMI and microbial ATP concentration in rat tissues. Since only a small amount of tissue was needed for the detection and the sample was not affected by self-decomposition, the method may extend the time range of PMI estimation.

Objective To evaluate the effect of CYP3A5 genetic polymorphisms on tacrolimus (Tac) concentration/dosing and other clinical outcomes in a pilot cohort of 113 Chinese HTx recipients.Methods Association between CYP3A5 genetic variants and blood dose-adjusted trough concentrations (C0/D) of Tac at 1st month at the beginning of the immunosuppressive therapy was evaluated in cohorts of 113 patients,then at 1st,3rd,6th,and 12th months after transplantation in 41 patients who received Tac-based immunosuppressive therapy and never changed within one year after transplantation,respectively.In addition,we also evaluated the association between CYP3A5 genetic variants and other clinical outcomes,such as the classifications of endomyocardial biopsy and longterm prognosis.Results The CYP3A5 wild homozygote (* 1/* 1),mutant homozygote (* 3/* 3),and mutant heterozygote (* 1/* 3) occurred in 5,34 and 74 recipients respectively.The gene mutation rate of CYP3A5 in this cohort of Chinese HTx recipients was 80.5 ％ and the homozygous proportion was 65.5％.Compared with CYP3A5 expressors (* 1/* 1 or * 1/* 3),CYP3A5 nonexpressors (* 3/* 3) had a higher Tac C0/D at 1st month (47.34 ± 11.40 vs.116.11 ± 42.40 vs.293.70 ± 171.20,P =0.000),as well as other studied time points (3rd month:98.32 ± 39.43 vs.292.07 ± 141.08,P=0.003;6th month:90.00 ± 21.31 vs.341.68 ± 165.02,P =0.002;and 12th month:96.02 ± 29.33 vs.339.23 ± 162.30,P =0.018);and might have a lower classification of endomyocardial biopsy at 1st month (1.43 ± 0.73 vs.1.50 ± 0.58,P =0.867),3rd month (1.55 ±1.00 vs.2.00 ± 1.73,P =0.512),and 6th month (1.36 ± 0.84 vs.2.33 ± 1.53,P =0.132);as well as a higher mortality due to acute organ rejection (10％ vs.0,P =0.244) and all-cause mortality (20％ vs.9.7％,P =0.580).Conclusion In Chinese HTx recipients,the frequency of this * 3 allele is lower than that has been reported in the white population.The determinations of CYP3A5 genetypes in heart transplant recipients are helpful to guide the individualized Tac regimens.

Objective To investigate the role of microRNA-21 (miRNA-21) in promoting proliferation of Kazakh's esophageal squamous cell carcinoma (ESCC) cell line Eca109 and Kazakh's ESCC tissues, and its effects on the expression of programmed cell death 4(PDCD4)gene.Methods TheEca109cellsweredividedintomiRNA-21Mimicsgroup(transfersequence:5′-UCAACAUCAGUCUGAUAAGCUA-3′),miRNA-21inhibitorgroup(transfersequence:5′-UAGCUUAUCAGACUGAUGUUGA-3′), negative control group (the transfer random sequence)and normal control group (no transfection).The cells were seeded at 4.5 × 105/well in 6-well cell culture plates. According to RNA interference technology,thecells were transfected with LipofectamineTM 2000. After transfection, the cell proliferation was determined by cell number counting.The expression of miRNA-21 was detected by real-time quantitative polymerase chain reaction (qRT-PCR), and the expression of PDCD4 protein was measured by Western blot.Meanwhile, the expression of miRNA-21 and PDCD4 protein were performed in 18 pairs of Kazakh's ESCC and corresponding adjacent normal esophageal mucosa.ResultsAt 48 hour after transfection,compared with normal control group, the cell number increased 36％ (P=0.002) in miRNA-21 Mimics group, decreased 28％ (P=0.002) in miRNA-21 inhibitor group, and did not change significantly in negative control group (P=0.515). At 48 hour after transfection, the relative expression of miRNA-21 in miRNA-21 Mimics group, miRNA-21 inhibitor group, negative control group and normal controlgroup was 0.37±0.10, 9.17± 1.08, 0.74±0.23 and 1.04±0.34,respectively, and the relative protein expression of PDCD4 was 1.47 ± 0.11, 0.61±0.09, 0.89 ±0.12 and 0.79±0.02 accordingly.Compared with normal control group, the relative expression of miRNA-21 decreased (P=0.031) and the relative protein expression of PDCD4 up regulated (P=0.001) in miRNA-21 inhibitor group, the relative expression of miRNA-21 increased (P=0.001) and the relative protein expression of PDCD4 down regulated (P=0.030) in miRNA-21 Mimics group,and there was no significant difference in negative control group (P=0.272 and 0.541).In 16 pairs of Kazakh's ESCC tissues, the expression of miRNA-21 was significantly higher in tumor tissues (0.11 ±0.09) than that of corresponding adjacent esophageal normal tissues (0.03 ± 0.03, P=0.001), while the relative protein expression of PDCD4 was significantly lower (0.92 ± 0.39) than that of corresponding adjacent normal esophageal tissues (1.57 ± 0.80, P=0.004). And the higher expression of miRNA-21, the lower relative protein expression of PDCD4 (r=-0.538, P=0.046).ConclusionmiRNA-21 may promoted the cell proliferation through inhibiting PDCD4 expression,which involved in the pathogenesis of Kazakh's ESCC.

Objective To explore the feasibility of extending liver blood perfusion cessation by ultrasound combining microbubbles.Methods The livers of 9 healthy rabbits were treated with a pulsed therapeutic ultrasound device,in presence of microbubbles.For quantification of liver perfusion,contrastenhanced ultrasonography was performed on 6 rabbits before treatment and at different time points of 0 min,30 min,60 min and 48 hours after treatment.Pathological examination of the treated livers was performed immediately after treatment on the other 3 rabbits.Results The liver blood perfusion almost vanished immediately after treatment,remained at a low perfusion level from 30 to 60 min,and completely recovered 48 hours later.The peak intensity dropped from ( - 51.88 ± 4.26)dB to ( - 62.53 ± 4.83)dB after treatment and rose up to ( - 52.00 ± 4.60) dB 48 hours later.The peak intensities before treatment and 48 hours after treatment were significantly higher than those of 0 min,30 min and 60 min time points after treatment( P ＜0.05).Pathological examination showed significant swelling of hepatocytes and hemorrhage around portal veins.Conclusions Microbubbles enhanced ultrasound can induce liver blood perfusion cessation for up to 1 hours.The mechanism could be swelling of hepatocytes and hemorrhage of portal track.