AIM: To examine the effects of cigarette smo ke extract (CSE) and lipo polysaccharide (LPS) on the production of transforming growth factor-? 1 (TGF- ? 1 ) mRNA and protein in cultured human embryonic lung fibroblasts (HELF). METHODS: The cultured HELF were incubated with CSE, LPS or CSE in com bination with LP S for 24 hours at 37℃, respectively. The total RNA was extracted from the cells . The expression levels of TGF-? 1 mRNA were evaluated by reverse transcription - p olymerase chain reaction (RT-PCR). The mRNA levels of TGF-? 1 were corrected by GAPDH transcripts, and TGF-? 1 protein levels were determined by immunocytoch emistry. RESULTS: CSE stimulated the TGF-? 1 mRNA expression i n HELF at lower concentr ati ons (1∶50 and 1∶25)( P 0.05). LPS enhanced TGF-? 1 mRNA levels in HELF at a ll three doses (0.1 mg/L, 1 mg/L, and 10 mg/L)( P

Objective　To investigate the expression of TGF-β and TGF-β receptor in human breast cancer cell Bcap-37 inhibited by soybean isoflavones. Methods　mRNA and protein of TGF-β1、TGF-βRⅠ in Bcap-37 cells were examined with reverse transcription ploymerase chain reaction(RT-PCR) and immunohistochemistry after cells were treated with daidein or genistein for 1-4 d.The expression of TGF-β1 and TGF-β2 was determined with TGF-β resistance test. Results　The TGF-β1, TGF-β2 and TGF-β recepor increased in Bcap-37 cells at a concentration of 3×10-5 mol/L of genistein. No changes was found when treated with daidzein. Conclusion　Genistein may inhibit the proliferation of Bcap-37 cells and accompany with increasing expression of TGF-β1, TGF-β2 and TGF-β receptor.

AIM:To observe the changes of nuclear factor-?B (NF-?B) activity and inducible nitric oxide synthase (iNOS) expression in hypoxic pulmonary hypertension(HPH). METHODS:The rat model of HPH was used. The NF-?B activity and iNOS expression in lung tissue were determined by immunohistochemistry(IHC), in situ hybridization (ISH), RT-PCR and Western blot.RESULTS:ISH showed that iNOS mRNA expression in intraacinar pulmonary arteriole (IAPA) in H28 d group(hypoxic treatment for 28 days) was stronger than that in normal group, H5d group and H14 d group. RT-PCR showed that the iNOS mRNA in H28 group was 2.1 times, 1.9 times and 1.8 times higher than that in normal group, H5d group and H14 d group, respectively. The nucleic anti-NF-?B stain was observed in H28 d group, which was significantly stronger, but the I-?B amount was 2.8 times, 2.7 times and 2.5 times lower than that in normal group, H5d group and H14 d group, respectively. CONCLUSION: The activity of NF-?B was correlated with the hypoxic pulmonary vessel structural remodeling and iNOS expression.

AIM: To investigate the effect of antisense oligonucleotides(ASON) of c-sis on cellular cycle and proliferation of pulmonary artery vascular smooth muscle cells(VSMC).METHODS: Tissue mass culture was done to get VSMC of pulmonary artery. Different concentrations of antisense oligonucleotides of c-sis were added into the cultures to observe the VSMC proliferation curve using MTT test. The changes of VSMC cellular cycle were also observed by flow cytometry. RESULTS: ASON with mid-to high concentrations restrained the proliferation of VSMC apparently with the peak of cell growth being attenuated or eliminated. Affected by mid-concentration ASON, PDGF-BB showed significant accelerating effect on the proliferation of VSMC. The ratio of G 0/G 1 in cellular cycle was increased significantly in VSMC culture with ASON in comparison with control. The G 0/G 1 ratio also showed significant differences among different concentration of ASON groups( P

AIM: To investigate the expression of phosphatidylinositol 3-kinase (PI-3K) during the proliferation of the hypoxic pulmonary arterial smooth muscle cells(PASMC). METHODS: The cultured PASMC was divided into two groups: serum-free medium (SFM) group and 48 hours hypoxia (H48h) group. Immunohistochmistry(IHC) and Flowcytometer(FCM) were used to detect the cultured PASMC. RESULTS: The positive expression of PI-3K p110 existed in both cultured PASMC groups. The staining intensity in H48h group (0 1891?0 0301) was significantly stronger than that in SFM group (0 1025?0 0164, P

In order to study the effect of nitric oxide (NO) on the expression of hypoxia-inducible factor-1 alpha (HIF-1alpha) mRNA in hypoxic pulmonary hypertension (HPH) rats, 30 healthy male Wistar rats were randomly divided into normoxic control group, chronic hypoxic group and hypoxia plus L-arginine (L-Arg) group. The animal model of HPH was developed. The mean pulmonary arterial pressure (mPAP) was measured by inserting a microcatheter into the pulmonary artery. The HIF-1alpha mRNA expression levels were detected by in situ hybridization (ISH) and semiquantitative RT-PCR. It was found that after 14 days hypoxia, the mPAP in normoxic control group (17.6 +/- 2.7 mmHg, 1 mmHg=0.133 kPa) was significantly lower than that in chronic hypoxic group (35.8 +/- 6.1 mmHg, t=0.2918, P<0.05) and mPAP in chronic hypoxic group was higher than that in hypoxia plus L-arginine group (24.4 +/- 3.8 mmHg, t=0.2563, P<0.05). ISH showed that the expression of HIF-1alpha mRNA in the intraacinar pulmonary arteriolae (IAPA) in normoxic control group (0.1076 +/- 0.0205) was markedly weaker than that in chronic hypoxic group (0.3317 +/- 0.0683, t=3.125, P<0.05) and that in chronic hypoxic group was stronger than that in hypoxia plus L-arginine group (0.1928 +/- 0.0381, t=2.844, P<0.05). RT-PCR showed that the content of HIF-1alpha mRNA in chronic hypoxic group (2.5395 +/- 0.6449) was 2.16 times and 1.75 times higher than that in normoxic control group (1.1781 +/- 0.3628) and hypoxia plus L-arginine group (1.4511 +/- 0.3981), respectively. It is concluded that NO can reduce the mPAP by the inhibition of the expression of HIF-1alpha mRNA, which may be one of the mechanisms through which NO affects the pathogenesis of HPH.
Anoxia/metabolism ; Arginine/pharmacology ; Hypertension, Pulmonary/*metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; Nitric Oxide/*pharmacology ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Random Allocation ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors/*biosynthesis ; Transcription Factors/genetics

Objective This study was to examine the expression of tyrosine kinase receptor(TKRs)C-kit,C-abl and PDGFR?,in ovary carcinoma.Methods The expression of C-kit,C-abl and PDGFR? in tumor tissue of 60 specimens of ovary carcinoma and normal fissue of 20 specimens of overy was examined by immunohistochemistry SP method.Results Immunoreactivity was detected in 79% of the tumor to at least one TKR.The total positive expression rate of C-kit,C-abl and PDGFR? in ovary carcinoma was 58.3%,70%,73.3%,respectively.The positive expression rate of C-kit and PDGFR? is significantly higher in tumor tissues than in normal tissues(P

The alpine plant Gentiana robusta is an endemic species to the Sino-Himalayan subregion. Also, it is one of the original plants used as traditional Tibetan medicine Jie-Ji. We sequence the nuclear ribosomal internal transcribed spacer (ITS) regions, matK, rbcL, rpoC1, trnL (UAA), psbA-trnH, atpB-rbcL, trnS( GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF( GAA) fragments of cp DNA in both G. robusta and such relative species as G. straminea, G. crassicaulis and G. waltonii. With Halenia elliptica as the outgroup, molecular systematic analysis reveals that G. robusta is a natural hybrid. G. straminea is the mother of hybrids, but the father is not very clear. In addition, the molecular markers for distinguishing G. robusta from the parental species or closely related species are identified, respectively. Our studies may provide valuable reference for the species identifications of medicinal plants with complex genetic backgrounds.
DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; Gentiana ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; classification ; genetics

Objective To study the short-term clinical outcomes of unicompartmental knee arthroplasty for medial compartmental knee osteoarthritis,and to compare 2 kinds of unicompartmental prosthesis.Methods From March 2010 to June 2013,data of 43 patients underwent unicompartmental knee arthroplasty (UKA) were retrospectively analyzed.17 patients (17knees) used rotating platform prosthesis,and 26 patients (28 knees) used fixed bearing prosthesis.There were 7 males (7 knees)and 9 females (10 knees) in rotating platform group,with an average age of 64.1 years (range,54-82 years);while 10 males (10knees) and 17 females (18 knees) in fixed bearing group,with an average age of 62.2 years (range,43-79 years).All patients presented signs of narrowed medial joint space,medial tenderness and pain on weight-bearing.X-ray and MRI were used for documenting joint narrowing and cartilage defect.The pain and the knee functions were recorded both pre and post-operatively with knee society score (KSS),2 cases of simultaneous anterior cruciate ligament (ACL) reconstruction were assessed with TegnerLysholm knee scoring scale as well.Results All 43 patients were followed up for 6 month to 37 months,and the average duration was 21.1 months.There were no dislocations,joint infection,deep venous thrombosis,prosthetic loosening,etc.The KSS in rotating platform group was 56.11 ±9.51 preoperatively,and 92.23±5.46 postoperatively.While the KSS in fixed bearing group was 57.11 ±9.56,and 93.69±6.37,respectively.There were statistical differences comparing between preoperative and postoperative KSS knee scores.There was no significant difference in KSS scores between rotating platform group and fixed bearing group.Conclusion Unicompartmental knee arthroplasty is a less invasive and effective method for knee osteoarthritis in medial compartment with less complications.There was no significant difference in clinical outcomes between rotating platform and fixed bearing design in terms of patients' satisfactory rate,clinical and functional outcomes in this short-term follow-up study.

To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.
Cell Line, Transformed ; Cloning, Molecular ; Embryo ; Eukaryotic Cells/*metabolism ; Gene Expression ; Genetic Vectors ; Kidney/cytology ; Kidney/*metabolism ; Peroxidases/*biosynthesis ; Peroxidases/genetics ; Plasmids/*genetics ; Transfection