Objective To explore the effect of RNA interference on the bionomics of Cathepsin L of hepatoma carcinoma cells. Methods In this study, the experimental group was the Cathepsin L RNA interference group. Control groups were the normal blank hepatoma carcinoma cell group (the blank group) and the Cathepsin LRNA interference blank group (the fluorescence control group). Observing times were ld,3d and 6d after RNA interference. Transfection efficiency in each group was observed. Expression of Cathepsin L of hepatoma cells was detected by immunofluorescence, RT-PCR and WB. Cell vigor was detected by MTT assay. Changes in the cell cycle and apoptosis were observed by flow cytometry. Transwell cabin was used to detect the changes of cell invasive power.Results In the experimental group, Cathepsin L mRNA level and protein level significantly decreased, the proliferation index significantly decreased and apoptosis index significantly increased. The invasive power also decreased. Conclusion RNAi interference can inhibit Cathepsin L expression, cell proliferation and cell invasive power efficiently.

Objective To investigate the molecular mechanism of epithelial-mesenchymal transition (EMT) induced by activated vascular endothelial growth factor receptor-1 ( VEGFR-I ) in cell line MHCC97-H.Methods MHCC97-H cells were cultured in DMEM with 1％ fetal bovine serum (control group),10 μmol/L PP2 (PP2 group),10 μmol/L PBS (PBS group),50 μmol/L VEGF-B (VEGF-B group),l0μmol/L PP2 and 50 μmol/L VEGF-B (PP2 +VEGF group),10 μmol/L PBS and 50 μmol/L VEGF-B (PBS + VEGF-B group),respectively.Protein expressions of epithelial marker E-cadherin,α-catenin and mesenchymal marker vimentin and N-cadherin were detected by Western blot.The expression sites of E-cadherin,α-catenin and mesenchymal marker vimentin and N-cadherin were detected by cell immunofluorescence.The ability of invasion and migration of cell line MHCC97-H were assessed by cell invasion and migration test.All data were analyzed by the t test.Results The expressions of E-cadherin,α-catenin,vimentin and N-cadherin were 3.23 +0.76,3.01 ±0.25,3.01 +0.22 and 2.63 +0.40 in the control group,4.18 +0.32,3.29 +0.11,4.85 +0.36 and 3.02 +0.52 in the PP2 group,2.83 +0.65,3.03 +0.27,1.37 ±0.24 and 2.98 ±0.36 in the PBS group,2.06 ±0.15,2.84 ±0.76,5.79 ± 0.38 and 5.54 ± 0.28 in the VEGF-B group,6.12 ± 0.08,5.45 ± 0.37,3.36 ± 0.42 and 3.26 ±0.13 in the PP2 + VEGF-B group and 1.36 ±0.54,1.26 ±0.45,4.05 ±0.17 and 1.05 ±0.33 in the PBS +VEGF-B group.There was a significant difference in the expressions of E-cadherin and α-catenin between the PP2 +VEGF-B group and the VEGF-B group (t =7.625,9.931,P ＜ 0.05 ).The expressions of vimentin and N-cadherin in the PP2 + VEGF-B group were significantly lower than those in the VEGF-B group (t =12.001,11.910,P ＜ 0.05).Six hours after the treatment with VEGF-B,the numbers of MHCC97-H migrated were 19 ± 1,5 ± 2and 16 ± 1 in the VEGF-B group,PP2 + VEGF-B group and PBS + VEGF-B group,respectively.The number of MHCC97-H cells migrated in the VEGF-B group was greater than that in the PP2 ± VEGF-B group ( t =13.566,P ＜ 0.05 ).The number of MHCC97-H cells passed through the Boyden chamber was 4 + 2,which was significantly less than 16 ± 1 of the VEGF-B group (t =12.350,P ＜0.05).Conclusion EMT induced by activated VEGFR-1 was mediated via c-Src kinase signal transduction in MHCC91-H cell line,and c-Src may be a potential target to interfere the invasion and migration of hepatic cancer cells.

In the present paper, we measured DNA content of uninuclear Plasrnodium berghei traced with DAPI by means of fluoromi-crospectrophotmeter. The results indicate that the DNA replication of parasite was continuous and it's content was high polyploidy and phase G1 of proliferating cycle was not evident. The dispersion degree of distribution of DNA content in P. R. was markedly lower and the peak sitewas more concentrated and obviously shifted to the right compared with P. N. It was suggested that the speed of DNA replication and proliferating vitality of parasites after producing resistance to SA markedly decreased, indicating that the changes of proliferating kinetics of P. R. happened.

Objective To investigate the protective effect of modified Qianjin Weijing Decoction(MQJWJD)on TNF-α and NF-κB in rats with lung injury induced by particulate matter; To discuss relevant mechanism of action. Methods A tracheal drip 15 mg/kg fine particles of saline solution was used to establish modeling, every other day, three times. Thirty-two Wistar rats were randomly divided into normal group, model group and MQJWJD high-dose and low-dose groups, with eight rats in each group. Medication groups were given relevant medicine for gavage. The level of TNF-α in bronchoalveolar lavage fluid (BALF) was measured by ELISA. The expression of NF-κB protein in lung tissues was measured by immunohistochemistry. The histopathology of the lung injury was observed by light microscope. Results Compared with normal group, the level of TNF-α and the expression of NF-κB protein in the model group were higher than those in the normal group (P<0.01). Compared with model group, the level of TNF-α and the expression of NF-κB protein in MQJWJD low-dose and high-dose groups were lower than those in the model group (P<0.05, P<0.01). Pathological observation showed that, compared with normal group, model group showed intratracheal, alveolar and interstitial bacteria within a large number of fine particles calm, alveolar and pulmonary interstitial visible large amounts of phagocytic fine particles of macrophages and accompanied by more neutrophils and lymphocyte infiltration; Lung tissue pathological changes were significantly lighter in MQJWJD high-dose and low-dose groups than the model group. MQJWJD high-dose group showed mild inflammation, alveolar and pulmonary interstitial visible phagocytic fine particles of macrophages, a small amount of neutrophils and lymphocyte infiltration. Conclusion MQJWJD can reduce the pulmonary injury in rats induced by particulate matter and has protective effects on the rat model through decreasing the levels of TNF-α and the expressions of NF-κB protein in injured lung tissues.

OBJECTIVETo study the clinical features and diagnosis of intrahepatic cholestasis of pregnancy (ICP).

METHODSDuring the last 10 years 1241 cases of ICP stayed in our hospital. Their clinical data were retrospectively reviewed.

RESULTS5.2% of all the maternity patients had ICP. It occurred more in winter and 3.5% of ICP occurred in multiple pregnancies. The recurrence rate of ICP was 30.2%. On the average, it occurred at gestational week 32.6. Skin pruritus was the characteristic manifestation and the presenting symptom in 1201 patients (96.8%). The other presenting features included elevated serum ALT and AST (2.3%), jaundice (8 patients), diarrhea (3 patients), deep yellow urine (2 patients) and right upper abdominal pain (1 patient). The serum transaminases levels were elevated, of which 60% were between 50-200 IU/L. Serum total bile acid (TBA) levels were elevated in 82.4% of the patients and bilirubin levels in 33.4%. The elevated bilirubin levels were 30 to 90 micromol/L in 85% of those patients with this condition, and it was never higher than 170 micromol/L.

CONCLUSIONThe basic diagnostic points of ICP are pruritus and abnormal liver function characterized by increased transaminases and TBA. Therefore paying attention to typical pruritus and other atypical features such as elevated serum transaminases, jaundice, diarrhea, deep yellow urine and right upper abdominal pain during antenatal care is important for an early diagnosis of ICP.

The distribution information of Lomatogonium rotatum. was collected by interview investigation and field survey, and 55 related environmental factors were collected, the habitat suitability study was conducted based on geographic information system (GIS) and maximum entropy model. The AUCs of ROC curve were both above 0.99, indicating that the predictive results with the maximum model were highly precise. The results showed that 13 major environmental factors have obvious influence on ecology suitability distributions of L. rotatum, including month average temperature of February et al., the suitable distribution areas are mainly concentrated in the east-central of Inner Mongolia, including Hexigten banner, Duolun county, Zhenglan banner et al., The zoning results basically coincide with the genuine producing areas, and further afford new suitable distribution areas, which can provide reference for L. rotatum's wild nursery and the siting of introduction and cultivation.
China ; Ecosystem ; Environment ; Gentianaceae ; growth & development ; Geographic Information Systems ; Rain ; Temperature

One DRV strain was isolated from Sika Deer brain and sequenced. Nine overlapped gene fragments were amplified by RT-PCR through 3'-RACE and 5'-RACE method, and the complete DRV genome sequence was assembled. The length of the complete genome is 11863bp. The DRV genome organization was similar to other rabies viruses which were composed of five genes and the initiation sites and termination sites were highly conservative. There were mutated amino acids in important antigen sites of nucleoprotein and glycoprotein. The nucleotide and amino acid homologies of gene N, P, M, G, L in strains with completed genomie sequencing were compared. Compared with N gene sequence of other typical rabies viruses, a phylogenetic tree was established . These results indicated that DRV belonged to gene type 1. The highest homology compared with Chinese vaccine strain 3aG was 94%, and the lowest was 71% compared with WCBV. These findings provided theoretical reference for further research in rabies virus.
Amino Acid Sequence ; Animals ; Base Sequence ; DNA, Complementary ; chemistry ; genetics ; Deer ; virology ; Genome, Viral ; Molecular Sequence Data ; Phylogeny ; Rabies ; virology ; Rabies virus ; classification ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Viral Proteins ; genetics

A sensitive high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method was established for the determination of eplerenone (EP) in human plasma. The plasma samples of EP were extracted with ethyl acetate and separated by HPLC on a reversed phase C18 column with a mobile phase of 10 mmol x L(-1) ammonium acetate water solution-methanol (30 : 70, v/v). EP was determined with electrospray ionization-mass spectrometry (ESI-MS) in the selected ion monitoring (SIM) mode. The calibration curves were linear over the range of 2-4 000 ng x mL(-1) for EP. The lower limit of quantification was 2 ng x mL(-1). The method has been successfully applied in the pharmacokinetic study of the EP tablets. The main pharmacokinetic parameters of EP after oral administration of 25 mg, 50 mg, 100 mg were as follows, t1/2: (4.9 +/- 2.1), (4.7 +/- 1.5), (5.9 +/- 1.2) h; AUC(0-infinity): (4 402 +/- 1 735), (8 150 +/- 2 509), (13 783 +/- 4 102) microg x h x L(-1); and MRT: (6.2 +/- 2.1), (6.6 +/- 1.3), and (7.2 +/- 1.6) h. Parameters of EP after oral administration of multiple doses of 50 mg were as follows, t1/2: (6.1 +/- 1.7) h; AUC(ss): (10 071 +/- 4220) microg x h x L(-1); MRT: (8.1 +/- 2.3) h; and DF: (3.2 +/- 1.0).
Chromatography, High Pressure Liquid ; methods ; Humans ; Spectrometry, Mass, Electrospray Ionization ; methods ; Spironolactone ; analogs & derivatives ; blood ; pharmacokinetics

The beta-glucosidase encoding gene bglA was cloned from Bacillus polymyxa 1.794. The bglA gene was inserted in expression vector pET28a(+) and transformed into Escherichia coli BL21 (DE3), finally the recombinant strain BL1979 was obtained. Induced by IPTG, the expression P-glucosidase activity reached to 24.7 IU/mL. The optimum temperature and optimum pH of the recombinant expression P-glucosidase in BL1979 were 37 degrees C and 7.0 respectively,the purity can reach to 92.7%. Analysis of the fusion protein by nondenaturing gradient gel electrophoresis, we found the fusion protein exists in dimmer, tetramer,hexamer and octamer, they all have hydrolase activity.
Bacillus ; enzymology ; Escherichia coli ; genetics ; Plasmids ; Recombinant Proteins ; biosynthesis ; isolation & purification ; beta-Glucosidase ; genetics ; isolation & purification ; metabolism

No abstract available.
Anesthesia* ; Intubation, Intratracheal* ; Vocal Cord Paralysis*