OBJECTIVETo investigate the effect of human serum albumin (HSA) on cell attachment of human gingival epithelial cells (HGE).

METHODSHGE were primary cultured with keratinocyte serum-free medium (KSFM) and dispase. The cultured cells were immunohistochemically stained by monoclonal anti-pan cytokeratin. MTT test was employed to investigate the influence of HSA on the cell attachment on polystyrene surface. The cell growth curve of HGE which were cultured in KSFM with 50 g/L HSA was observed.

RESULTSThe results showed significant decrease in cell numbers within 8 hours after HGE were inoculated, in which the polystyrene surface was preincubated with 50 g/L HSA. But it did not prove to be the case from 10 hours to 24 hours after HGE were inoculated. There were no significant difference within 24 hours in cell numbers between cultured in KSFM with 50 g/L HSA and control. The cell numbers in cell growth curve of HGE in KSFM with and without 50 g/L HSA did not show significant difference.

CONCLUSIONSHSA preincubation on polystyrene were produce inhibitory effect of HGE attachment in early stage.

Cell Adhesion ; drug effects ; Cell Count ; Cell Division ; drug effects ; Cells, Cultured ; Epithelial Cells ; cytology ; drug effects ; Gingiva ; cytology ; drug effects ; Humans ; Polystyrenes ; Serum Albumin ; pharmacology

OBJECTIVETo investigate the effects of different concentrations of Gubishu containing serum on the proliferation of rabbit articular chondrocytes cultured in vitro.

METHODSArticular chondrocytes were obtained from the cartilage of 1-month rabbit and cultured in vitro. They were randomly divided into 8 groups,blank and Gubishu groups in different concentrations (5%, 10%,15%, 20%), MTT assay method was adopted to observe the influence of Gubishu containing serum with different concentrations to the proliferation of chondrocytes after incubated 1, 3, 5, 7 and 9 days.

RESULTSThe proliferation of chondrocytes was dependent on the concentration in Gubishu groups. At same time point,there was significant value between every groups, 20% concentration was greatest (P<0.05); There was significant differences between 5%, 10% and 20% concentration of the blank groups at same time point (P<0.05), and was not between 15% and 20% concentration at the 1, 3, 5 and 7 days (P>0.05), 20% concentration of the blank group was greatest. 20% concentrations of Gubishu containing serum was significantly greater than 20% concentrations of blank group at the 1, 3, 5 and 7 days (P<0.05).

CONCLUSION20% concentrations of Gubishu containing serum can significantly increase the proliferation of chondrocytes, and bring the logarithmic growth period forward to the 3 day.

Animals ; Cartilage, Articular ; cytology ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chondrocytes ; drug effects ; physiology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Male ; Rabbits ; Serum

OBJECTIVETo observe the effect of Guizhi plus Gegen Decoction (GGD) on ultrastructural changes of intervertebral disc annulus fibrosus cells.

METHODSRats' intervertebral disc annulus fibrosus cells were isolated and cultured using adherence wall screening method. After annulus fibrosus cells were intervened by GGD, the microstructure and ultrastructural features of untreated annulus fibrosus cells and annulus fibrosus cells treated by GGD containing serum at different concentrations were observed under optical microscope and electron microscope.

RESULTSUnder optical microscope, most annulus fibrosus cells showed irregular polygons and few in star shape with rich superficial ecphyma. The nuclei were oval, large and complete. Under electron microscope, most cells in the blank group were oval after intervened by GGD containing serum at different concentrations. The nucleus was large, deviated, and irregular, the heterochromatin scattered diffusely, partial mitochondria vacuolized, and rough endoplasmic reticulum dilated. In the low dose GGD group, increased mitochondria and condensed density could be seen. The rough endoplasmic reticulum were expanded, lipid drops or glycogen could be occasionally seen. In the middle dose GGD group, increased endoplasmic reticulum expansion and condensed density could be seen. More medium density protein sediment could be seen. Increased mitochondria with condensed density could be seen, showing irregular cystic form with various sizes nucleus. In the high dose GGD group, increased rough endoplasmic reticulum with obvious expansion could be seen. More high density protein sediment could be seen. The nuclei were deviated. More mitochondria could be seen with secretory granules in them.

CONCLUSIONSAfter intervened by GGD containing serum at different concentrations, the ultrastructure of annulus fibrosus cells were manifested as follows: (1) The endoplasmic reticulum increased more in the middle and high dose GGD groups than in the blank group and the low dose GGD group. Greater density protein sediment occurred, especially in the high dose GGD group. (2) GGD played an important role in preventing ultrastructural changes induced by the degeneration of annulus fibrosus cells.

Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Intervertebral Disc ; cytology ; drug effects ; ultrastructure ; Rats ; Serum

OBJECTIVETo study relatively pharmacological activities of cinnamon acid in blood serum of rabbit administered cinnamon acid, cinnamon and Jingui Shenqi pills.

METHODRP-HPLC determine and analysis blood serum sample from rabbits administered cinnamon acid, cinnamon and Jingui Shenqi pills. Condition of colour spectrum was Symmetry C18 (3.9 mm x 150 mm, 5 microm) chromato bar, mobile phase was methanol-1% glacial acetic acid water-solution (45:55), flow rate was 0.8 mL x min(-1), temperature of bar was 35 degrees C, detection wave length was 285 nm. The serum pharmacokinetic parameters were calculated with 3p87.

RESULTLinear range of cinnamon acid is from 0.06-15 microg x mL(-1) (r = 0.9997), the lowest detectability is 0.054 microg x mL(-1). Pharmacokinetic process of cinnamon acid in rabbit could be all fitted to two-compartment model.

CONCLUSIONSensitive and exclusive HPLC that adopt can exactly detect serum concentration in rabbits administerd cinnamon acid. Pharmacokinetic parameters of three conditions can reveal pharmacokinetics regularity of cinnamon acid in rabbit.

Animals ; Cinnamomum zeylanicum ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Male ; Rabbits ; Serum ; chemistry ; drug effects

OBJECTIVETo study the effects of Bushen Zhuangjin Decoction (BZD) containing serum on the apoptosis of chondrocytes induced by mechanics stimulus.

METHODSThe BZD containing serum was extracted. The chondrocyte nutritive media was divided into 3 groups, i.e., the common nutritive medium group, the blank rabbit serum medium group, and the BZD nutritive medium group. The apoptosis of chondrocytes was induced by continuing mechanics stimulus in 24 h. Then the chondrocytes were collected. The apoptosis rate of chondrocytes was determined by flow cytometry. The contents of interleukin 1beta (IL-1beta) and nitric oxide (NO) in the corresponding media were determined.

RESULTSThe apoptosis of chondrocytes in the BZD nutritive medium group (19.55 +/- 7.98)% was lower than that of the common nutritive medium group (39.32 +/- 13.45)% and the blank rabbit serum medium group (37.87 +/- 9.67)%, showing statistical difference (P < 0.05). The contents of IL-1beta and NO were also lower in the BZD nutritive medium group with statistical difference when compared with those of the other two groups (P < 0.05).

CONCLUSIONBZD containing serum could protect mechanics stimulus induced apoptosis of chondrocytes.

Animals ; Apoptosis ; drug effects ; Cartilage, Articular ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Interleukin-1beta ; analysis ; Nitric Oxide ; analysis ; Rabbits ; Serum

OBJECTIVETo study influence of serum containing Liuwei Dihuang decoction (see text) on proliferation and differentiation of osteoblast form neonatal SD rats cultured in vitro at different times and different stretch stress.

METHODSAfter osteoblast cultured for 24 hours in the serum containing Liuwei Dihuang decoction (see text) and serum in control group, the 0.5 Hz frequency, 6% and 12% stretch-stress were added. The MTT1 and the activity of ALP were measured at the 12th and 24th hours, and the data were analyzed.

RESULTS1. In the environment of stretch stress to the frequency of 0.5 Hz, and stretched for 24 hours, the osteoblast was stimulated under elongation rate of 6% and 12%; the proliferation and differentiation of osteoblast was more active under elongation rate of 12% than that of 6%. 2. There were no stimulating effects on osteoblast proliferation and differentiation of serum containing Liuwei Dihuiang decoction (see text) acted on osteoblast cells of SD rats cultured in vitro for a shot time.

CONCLUSIONStretch stress environment can enhance osteoblast proliferation and differentiation cultured in vitro.

Alkaline Phosphatase ; metabolism ; Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Male ; Osteoblasts ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Serum ; Stress, Mechanical

The activation and proliferation of hepatic stellate cells (HSCs) is the central link in the formation and progression of liver fibrosis. From the research of serum pharmacological method of Chinese material medica, we found Chinese material medica serum could interfere in the formation and progression of liver fibrosis. This paper reviewed the progress on Chinese material medica serum and HSCs. The authors pointed out Chinese material medica serum could effect on the activation, proliferation, contraction and apoptosis of HSCs, which probably is an effective therapy for liver fibrosis.
Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; cytology ; drug effects ; Humans ; Liver Cirrhosis ; prevention & control ; Serum

The purpose of the present study was to investigate the effect of advanced glycated albumin (AGE-alb) on pyroptosis of macrophages and the underlying molecular mechanisms. RAW264.7 macrophages were treated with AGE-alb (1, 2, 4 and 6 g/L) and control albumin (C-alb, 4 g/L) for 24 h, or preincubated with MCC950 (1 μmol/L) for 1 h and then treated with AGE-alb (4 g/L) for 24 h. Cell viability and caspase-1 activity were measured by MTT and assay kits, respectively. Lactate dehydrogenase (LDH) activity and the levels of interleukin-1β (IL-1β) and IL-18 in media were detected. Cell death degree was evaluated by TUNEL and Hoechst 33342/PI staining. The protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), procaspase-1 and cleaved caspase-1 were assessed by Western blot. The results showed that AGE-alb treatment caused obvious decrease in cell viability and increases in LDH leakage and the percentages of TUNEL- or PI-positive cells in a concentration-dependent manner. Additionally, AGE-alb promoted IL-1β and IL-18 secretion, upregulated NLRP3 expression, and increased caspase-1 activity especially at the dose of 4 and 6 g/L. However, MCC950 (an NLRP3 inhibitor) pretreatment inhibited significantly the decrease in cell viability and the increases in LDH leakage and percentages of TUNEL- or PI-positive cells induced by AGE-alb. Furthermore, MCC950 attenuated obviously AGE-alb-induced IL-1β and IL-18 secretion and caspase-1 activation. These results indicate that AGE-alb may induce macrophage pyroptosis, and the mechanism is at least partially by activating NLRP3-caspase-1 pathway.
Caspase 1 ; Gene Expression Regulation ; drug effects ; Interleukin-1beta ; genetics ; Macrophages ; drug effects ; NLR Family, Pyrin Domain-Containing 3 Protein ; genetics ; Pyroptosis ; drug effects ; Serum Albumin ; pharmacology

OBJECTIVEImmunsuppressive therapy is a major therapy for severe aplastic anemia, and antithymocyte /antilymphocyte globulin (ATG/ALG) is usually used. This study investigated the therapeutic effect of ATG/ALG on severe aplastic anemia and explored the management of therapy-related complications.

METHODSClinical data of 28 children with severe aplastic anemia who received ATG/ALG treatment from December, 1994 through to September, 2005 were analyzed retrospectively.

RESULTSOf the 28 patients, 2 were nearly cured (7.1%), 4 were relieved (14.3%) and 12 were improved (42.9%) based on a hemoglobin/white blood cell/platelet count. These results represented an overall effective rate of 64.3%. Clinical evidence of serum sickness developed in 19 patients, manifesting as fever (n = 9), cutaneous eruptions (n = 12), arthralgias (n = 7), myalgia (n = 7) and arthrocele (n = 3). Serum sickness occurred 5-17 days after ATG/ALG administration and lasted for 1-15 days (mean 4.4 days). Three children with mild serum sickness symptoms recovered without any treatment. The symptoms of the other 16 patients disappeared after 3-5 days of methylprednisolone treatment (10 mg/kg daily). However, 3 patients had relapses at 2-4 days after termination of methylprednisolone therapy. Another course of methylprednisolone therapy was administered to the 3 patients until the symptoms disappeared. The patients with no serum sickness or with mild serum sickness had a better response to ATG/ALG therapy than those who had severe serum sickness (100% vs 60%; P < 0.05).

CONCLUSIONSATG/ALG therapy for severe aplastic anemia is effective. Serum sickness is a common complication in children with severe aplastic anemia following ATG/ALG therapy, but can be improved by methylprednisolone application.

Anemia, Aplastic ; drug therapy ; Antilymphocyte Serum ; adverse effects ; therapeutic use ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Serum Sickness ; etiology ; T-Lymphocytes ; immunology

Immune complex deposits have been found in the choroid plexus in patients with systemic lupus erythematosus, and it can be assumed that an immune complex injury to the choroid plexus might be related to the neuropsychiatric disorder seen in patients with SLE. Acute serum sickness was experimentally induced in rabbits by intravenous injection of crystalized BSA. Prednisolone in conventionl dosage was administered to study the immunologic injury of the choroid plexus as well as the mechanisms involved in the prednisolone effect. Light, electron microscopic and immunofluorescent studies were made. The host immunoglobulins(IgG, IgA, IgM) and beta 1 C globulin were demonstrated in the choroid plexus. Histopathological findings included mild to moderate interstitial and perivascular lymphocyte and plasma cell infiltrations and edema. Control animals showed no immune deposits and no histopathologic changes. Electron microscopic findings comparing the immunofluorescent and histopathologic changes were minimal, and showed sparse, vague electron dense deposits particularly in the interstitial spaces, knob-like focal thickening of vascular basement membrane, swelling of endothelial cells, and some accentuation of interstitial cells. The morphologic and functional similarities of the choroid plexus and glomerular basement membrane, the findings in morphologic, electron microscopic and immunofluorescent examinations of the experimental rabbits, along with the observed effects of prednisolone, together with similar reports in the recent literature suggest that immunologic injury of the choroid plexus could be considered as a new disease entity. This immunologic injury might play a significant role in neuropsychiatric disorders in the long standing immune complex deposit diseases. The very interesting finding is the nature and function of the interstitial cell between the endothelial (vascular) and epithelial side basement membranes, and speculation as to whether or not the role of this interstitial cell in choroid plexus injury may be in its possible analogy with glomerular mesangial cells.
Acute Disease ; Animal ; Choroid Plexus/drug effects ; Choroid Plexus/immunology* ; Choroid Plexus/pathology ; Lupus Erythematosus, Systemic/etiology ; Prednisolone/pharmacology* ; Rabbits ; Serum Sickness/chemically induced ; Serum Sickness/complications* ; Serum Sickness/immunology