In the study, the effects of Panax notoginseng saponins (PNS) on alveolar epithelial to mesenchymal transition (EMT) and extracellular matrix degradation were observed in a type of human alveolar epithelial cell, A549 cells, stimulated by TGF-beta1. Firstly, MTT method was applied to evaluation of cellular proliferation and found that PNS from 12.5 mg x L(-1) to 200 mg x L(-1) dosage could not inhibit significantly cellular proliferation. Then, cells were divided into five groups, normal group, TGF-beta1 group, TGF-beta1 + 50 mg x L(-1) PNS group, TGF-beta1 + 100 mg x L(-1) PNS group and TGF-beta1 + 200 mg x L(-1) PNS group. Normal cells were not stimulatec by TGF-beta1; TGF-beta1 cells were only stimulated by TGF-beta1 and the other cells were stimulated by TGF-beta1 with different doses of PNS, respectively. After stimulation, cells and supernatants were collected for assays. Cellular roundness was applied to quantitative evaluation of morphological change. Immunocytochemistry was applied to examine E-cadherion, a-SMA and FN proteins expression in the cells. Enzyme linked-immunosorbent assay was applied to MMP-9 and TIMP-1 levels. The results showed that EMT of A549 cells was induced by TGF-beta1, showing significant change of roundness, E-cadherion, alpha-SMA and FN (P < 0.05, P < 0.01). Compared to TGF-beta1, PNS significantly inhibited the changes of roundness (P < 0.05), FN and alpha-SMA (P < 0.05, P < 0.01) and not significantly inhibited the change of E-cadherion. Furthermore, MMP-9 levels were significantly increased by TGFbeta1 stimulation (P < 0.05), without significant change of TIMP-1. Compared with TGF-beta1, PNS could significantly increase MMP-9 level (P < 0.05) and decrease TIMP-1 levels (P < 0.05, P < 0.01). In conclusion, PNS could inhibit alveolar epithelial cell EMT induced by TGF-beta1, with increase of extracellular matrix degradation ability, which showed anti-fibrosis of lung ability.
Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; Panax notoginseng ; chemistry ; Pulmonary Alveoli ; cytology ; drug effects ; metabolism ; Saponins ; pharmacology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo observe the effects of oxymatrine on the expression of interleukin-6 (IL-6), interleukin-1beta (IL-1beta) mRNA of human periodontal ligament cell (PDLC) stimulated by lipopolysaccharides (LPS), and to discuss oxymatrine's inhibition mechanism on periodontal inflammation stimulated by LPS.

METHODSFirstly, isolate PDLC externally and culture them; for oxymatrine experimental group we used different combination of LPS and oxymatrine in different concentration, and for the matched group we use DEME nutrient solutions of 1% FBS. Then reverse transcription-polymerase chain reaction (RT-PCR) for checking the level of IL-6 and IL-1beta mRNA.

RESULTS25 microg x mL(-1) LPS can significantly enhance the expression of both IL-6 and IL-1beta mRNA's level, and oxymatrine could restrain above phenomena.

CONCLUSIONOxymatrine can restrain the expression of IL-6 and IL-1beta mRNA of human PDLC stimulated by LPS.

Alkaloids ; Humans ; Interleukin-1beta ; Interleukin-6 ; Lipopolysaccharides ; Periodontal Ligament ; Quinolizines ; RNA, Messenger

Standardized Manipulation of Acupuncture and Moxibustion Part 8: Intradermal Needle was compiled with the following principles. The compiling standard, technical features and clinic manipulations of intradermal needle were taken as the basic principle for compiling. Literature research, expert survey and clinic practice verification were applied as the drafting methods. The key issues were focused on the relationship between standardization and individualization, normalization and effectiveness, qualification and quantification. And the postural selection, reinforcing and reducing manipulations, fixing materials and embedding duration involved in intradermal needling were emphasized particularly. At the same time, details and the future way of thinking of intradermal needle were expounded in this article as well.
Acupuncture Therapy ; instrumentation ; standards ; China ; Humans ; Moxibustion ; standards ; Needles ; standards ; Reference Standards

OBJECTIVETo construct the adenoviral vector containing human Bcl-2 gene and to study the expression of the gene in the spiral ganglion cells (SGC) in vitro.

METHODSHuman Bcl-2 cDNA obtained from the plasmid pUC-CAGGS/Bcl-2 was cloned into the plasmid pAdTrack-CMV. Then, pAdTrack/Bcl-2 was cotransferred with adenoviral backbone vector into E. coli strain BJ5183. The recombinant adenoviral plasmid was identified by restriction analysis with Pac I and transfected into HEK293 cells to package and amplify recombinant adenovirus particles which would be identified by Electron microscope. After the adenovirus infected the rat spiral ganglion cells, the expression of Bcl-2 gene was detected by Western Blot and RT-PCR.

RESULTSThe recombinant AdEGFP/Bcl-2 plasmid was correctly constructed and confirmed by restriction endonuclease analysis. The viral particles in HEK293 cells were identified by Electron microscope. RT-PCR and Western blot showed that Bcl-2 gene was exactly transcripted and expressed in transgene SGC.

CONCLUSIONSThe method based on homologous recombination in bacteria is simple and high efficient. The recombinant adenoviral vector containing human Bcl-2 cDNA was constructed and the transgene SGC expressed human Bcl-2 gene in vitro successfully. It provided foundation for the further study of protection for the impaired SGC by hBcl-2 gene.

Adenoviruses, Human ; genetics ; Animals ; Cells, Cultured ; Genes, Homeobox ; Genes, bcl-2 ; Genetic Vectors ; Humans ; Rats ; Rats, Sprague-Dawley ; Recombination, Genetic ; Spiral Ganglion ; metabolism ; Transfection ; Transgenes

OBJECTIVETo establish a new method on stratification analysis when the stratification limits of confounding factors was not clear or contradictory.

METHODData on a study of diabetes mellitus in Guangdong province collected in the year of 1997 and 1998 was analyzed using cluster-stratification analysis.

RESULTSThe efficiency of stratification analysis was improved and the confounding bias was effectively controlled with information bias avoided when the clusters-stratification analysis was applied.

CONCLUSIONThe problem was logically solved using cluster analysis as an assistant stratification means.

Adult ; Age Factors ; Aged ; Bias ; China ; epidemiology ; Cluster Analysis ; Confounding Factors (Epidemiology) ; Data Interpretation, Statistical ; Diabetes Mellitus ; epidemiology ; Epidemiologic Methods ; Humans ; Middle Aged ; Multivariate Analysis ; Reproducibility of Results ; Risk Factors

Purpose:To study the diagnostic value of using ultrasonography(US)、plain film of the abdomen(KUB)、 intravenous urography (IVU) in acute ureteral calculous obstruction. Methods: The routine B-US、diuretic US、 KUB、 IVU were retrospectively analyzed in 352 cases of acute ureteral calculous obstruction.Results: Of all the patients, 83.2 % (293/352) 、 96.6 % (340/352)were diagnosed as hydronephrosis by routine B-US、 routine B-US plus diuretic US, respectively. The hydronephrotic diagnosis of 59 suspected patients were79.7% (47/59)using diuretic US. The ureteral calculous dignosis of US、KUB、US plus KUB were 76. 1 % (268/352 )、81.6(284/348)and 91.8(323/352),respectively. Conclusion:The routine B-US is an effective method todiagnose acute upper urinary tract obstruction,and routine B-US plus diuretic US is more valuable. B-US plusKUB is the first approach to diagnose ureteral calculi.

This study was purposed to investigate the B cell-activating factor belonging to the TNF family (BAFF) and a proliferation-inducing ligand (APRIL) levels in bone marrow, and the BAFF receptor expression level on B cells in multiple myeloma (MM) patients, in order to explore the characteristics of B cells in bone marrow of MM patients. MM patients were studied before treatment (newly diagnosed group, 19 patients) and after treatment with improvement (stable group, 17 patients), 10 non-hematologic patients were selected as control (control group). The BAFF receptors (BAFF-R) and transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) on B cell (CD19(+)), naive B cell (CD19(+)IgD(+)) and memory B cell (CD19(+)CD27(+)) of bone marrow in all groups were detected by flow cytometry. The BAFF, APRIL level in bone marrow supernatant were tested with ELISA. The results showed that the BAFF-R expression level on CD19(+) cells in newly diagnosed group were higher than that in stable group and control group; there was no significant difference between the BAFF-R expression level on CD19(+)IgD(+) cells in newly diagnosed group and stable group, but BAFF-R expression level on CD19(+)IgD(+) cells in newly diagnosed group was higher than that in control group; the BAFF-R expression level on CD19(+)CD27(+) cells in newly group was higher than that in stable group and control group; there was no significant difference between the BAFF-R expression level on CD19(+) cells, CD19(+)IgD(+) cells or CD19(+)CD27(+) cells in stable group and control group. There was no significant difference among the TACI expression level on CD19(+) cells, CD19(+)IgD(+) cells or CD19(+)CD27(+) cells in newly diagnosed group, stable group and control group. The bone marrow supernatant BAFF level in newly diagnosed group was higher than that in stable group and control group, but there was no significant difference between stable group and control group. There was no significant difference among the bone marrow TACI levels in newly diagnosed group, stable group and control group. It is concluded that both the bone marrow BAFF level and the BAFF-R expression level on CD19(+) cell, CD19(+)IgD(+) cells and CD19(+)CD27(+) cells in MM patients increase, which may help to stimulate B cells, thereby may relate with to MM pathogenesis.
B-Cell Activating Factor ; metabolism ; B-Cell Activation Factor Receptor ; metabolism ; Bone Marrow ; metabolism ; Humans ; Middle Aged ; Multiple Myeloma ; metabolism ; pathology

OBJECTIVETo evaluate the in vivo and in vitro stability of (131)I-Herceptin and its form of existence in the blood.

METHODSHerceptin was labelled with iodine-131 using the Iodogen method. (131)I-Herceptin was stored at 4 degrees celsius for 3, 24, 48, 72 and 96 h, and the radiochemical purity (RCP) was measured by high performance liquid chromatography (HPLC). Five rabbits received injections of (131)I-Herceptin and at 1, 3, 6, 24, 48, 72, 96 and 120 h after the injection, blood samples were taken to measure the RCP of (131)I-Herceptin in the serum, and the radio count of the serum and blood cells was calculated.

RESULTSThe baseline RCP of (131)I-Herceptin was (94.9±2.7)%. The RCP was stable after placement at 4 degrees celsius for not over 72 h (F=15.985, P<0.001), but was significantly lowered to (82.6±2.8)% after preservation for over 72 h (t=9.971, P<0.001). Within the time of 1.0 to 96 h after injection in rabbits, (131)I-Herceptin existed mainly in the serum with a radio count of 81%-87%; 24 h after the injection, the RCP of (131)I-Herceptin in the serum was significantly lowered to (75.4±3.9)% (t=6.564, P<0.001).

CONCLUSIONStorage at 4 degrees celsius for no more than 72 h does not obviously affect the activity of (131)I-Herceptin in terms of RCP. After injection in rabbits, (131)I-Herceptin exists mainly in the serum and its radiochemical purity remains stable within 24 h, after which obvious degradation occurs.

Animals ; Antibodies, Monoclonal, Humanized ; pharmacokinetics ; Blood ; metabolism ; Cell Line, Tumor ; Drug Stability ; Humans ; Iodine Radioisotopes ; pharmacokinetics ; Rabbits ; Radiopharmaceuticals ; pharmacokinetics ; Trastuzumab

OBJECTIVETo investigate the correlation between auto-immune antibodies and rheumatoid arthritis (RA) patients of Shen deficiency syndrome (SDS), thus providing clinical evidence for further researches on molecular biological mechanisms of RA patients of SDS.

METHODSTotally 451 RA patients were assigned to the SDS group and the non-SDS group. Their general conditions (including gender, age, duration, and age of onset), C reactive protein (CRP), erythrocyte sedimentation rate (ESR), platelet (PLT), disease activities (DAS28), auto-antibodies [rheumatoid factor (RF), anti-CCP antibodies, anti-nuclear antibody (ANA)] were compared between the two groups.

RESULTS(1) The scores for EST, PLT, and DAS28 were obviously higher in the SDS group than in the non-SDS group (P < 0.05, P <0. 01). (2) The level of average RF was (697.32 +/-1 061.38 IU/mL) in the SDS group, higher than that in the non-SDS group (439.91 +/- 672.24 IU/mL, P <0.01). There was no statistical difference in anti-CCP antibody between the two groups (P >0.05).(3) The ANA positive rate of RA patients was 29. 63% (120/405). It was 37.19% (74/199) in RA patients of SDS and 22. 33% (46/206) in RA patients of non-SDS, showing statistical difference between the two groups (P <0.01). (4) The odds ratio for high level RF positive and ANA positive was 1. 574 and 2. 059 folds in RA patients of SDS as high as that in RA patients of non-SDS.

CONCLUSIONSRA patients of SDS would have higher risk of having auto-immune antibodies, fastened development, more worsen joint damage, and more poor prognosis. Its mechanisms might be closely associated with autoimmune tolerance.

Adolescent ; Adult ; Aged ; Arthritis, Rheumatoid ; blood ; diagnosis ; Autoantibodies ; blood ; Child ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Young Adult