BACKGROUND:Phytohemagglutinin (PHA) can stimulate the peripheral blood mononuclear cells (PBMCs) into cellcycle, and cause their immune activation, which is a common immune proliferation model. However, the role of non-PBMC ingredient of peripheral blood is unclear, as wel as the expression of endothelial cells related cytokines. OBJECTIVE:To study the effect of whole blood culture and PBMCs alone culture with PHA on the PBMC proliferation and apoptosis, expression of inflammatory cytokine and endothelial cellsecreted cytokine markers. METHODS:Morphological changes of PBMCs separated from normal karyotype human peripheral blood individual y cultured with or without PHA were observed. The PBMCs were col ected by whole blood culture or PBMC separated culture. mRNA was extracted for the fluorescence quantitative RT-PCR, which was applied to detect the cellproliferation, apoptosis, and expression of inflammatory cytokine and endothelial cellsecreted cytokines. The statistic analysis was used for the significance explication. RESULTS AND CONCLUSION:PBMCs alone cultured ere different from those undergoing whole blood culture. The PHA could up-regulate the gene expression of Ki67, proliferating cellnuclear antigen, Caspase 3, interferon-γ, tumor necrosis factor-βand interleukin-6, but down-regulate Protein C. This indicted that PHA could promote the proliferation and apoptosis of PBMCs and up-regulate the expression of inflammatory cytokines, but down-regulate the expression of endothelial cells secreted coagulation cytokines.

AIM: We hypothesized that PPAR? ligands stimulate endothelial-derived nitric oxide(NO) release to protect the vascular wall.Thus,the purpose of this study is to investigate the effects of ciglitazone(Cig) and fenofibrate(Fen) on angiotensin Ⅱ(AngⅡ)-induced decrease in endothelial NO synthase(eNOS) expression and NO production in human umbilical vein endothelial cells(HUVECs).METHODS: HUVECs were preincubated for 24 h with Cig(10~(-7), 10~(-6),10~(-5),10~(-4) mol/L) or Fen(10~(-5) and 10~(-4) mol/L),then incubated for 12 h with 10~(-7) mol/L AngⅡ.Total RNA was extracted,and the expression of mRNA and protein of eNOS was assessed by RT-PCR and Western blotting.NO production was measured by Griees method.RESULTS: In the presence of 10~(-7) mol/L AngⅡ for 12 h,NO production in cultured HUVECs was decreased(P

BACKGROUND:Hidden blood loss is an important risk for the intertrochanteric fracture patients, especial y the elderly patients, which can cause anemia in patients after internal fixation and can affect wound healing and patient recovery. OBJECTIVE:To compare the perioperative hidden blood loss and the risk factors of proximal femoral anti-rotation intramedul ary nail internal fixation and dynamic hip screw fixation for the treatment of femoral intertrochanteric fracture. METHODS:We selected 70 cases of femoral intertrochanteric fracture patients who treated with proximal femoral anti-rotation intramedul ary nail and dynamic hip screw fixation, including 21 patients with the age ≥ 80 years and 49 patients with the age<80 years;28 patients with the body mass index>30 kg/m2 and 42 patients with the body mass index ≤ 30 kg/m2;30 patients received anti-rotation intramedul ary nail internal fixation and 40 patients received dynamic hip screw fixation. The perioperative blood loss was calculated with Gross formula according to the changes of height, body mass index and the hematocrit before and after fixation. RESULTS AND CONCLUSION:The mean total blood loss was 936 mL, the mean dominant blood loss was 237 mL and the mean hidden blood loss was 699 mL. The hidden blood loss was accounted for 74.7%in total blood loss. The dominant blood loss in the dynamic hip screw fixation group was higher than that in the anti-rotation intramedul ary nail internal fixation group, and the hidden blood loss was lower than the anti-rotation intramedul ary nail internal fixation group. The total blood loss and the hidden blood loss of the elderly patients were higher than those of the non-elderly patients;there was no significant difference between male and female patients, obesity and normal patients. The results indicate that hidden blood loss is the major reason for total blood loss of femoral intertrochanteric fracture after internal fixation. The hidden blood loss of anti-rotation intramedul ary nail internal fixation is larger than that of dynamic hip screw fixation, and elder is the risk factor for hidden blood loss.

This paper describes a calibration phantom system for QCT bone mineral density determination, which consists of 4-standard-solid-sample calibration phantom, a quality assurance (QA) phantom and the bone mineral density analysis software. The system adds to the new applications of CT systems, and provides a new method with a good accuracy and reliability for the examination, diagnosis, prevention, treatment of osteoporosis diseases and the observation of curative effect of drugs.
Absorptiometry, Photon ; instrumentation ; methods ; Algorithms ; Animals ; Bone Density ; Calibration ; Equipment Design ; Humans ; Image Processing, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; methods ; Osteoporosis ; diagnostic imaging ; Phantoms, Imaging ; Software ; Tomography, X-Ray Computed ; instrumentation ; methods

Objective To compare the content of main constituents of Compound Wurenchun Capsules dissolved in serum and plasma of rats. Methods Serum and plasma containing drug were prepared after ig the preparation to rats. Lichrosphere C_ 18 (250 mm?4.6 mm, 5 ?m) column and Phenomenex Description C_ 18 (4.0 mm?3.0 mm) protective column were used. The mobile phase consisted of methanol-water, eluted in gradient mode. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and detection wavelength was 254 nm. Results The linear ranges of schisandrin and schisandrin B were 0.051 2 .614 4 and 0.039 8—0.477 6 ?g. The average relative recoveries of schinsandrin and schisandrin B were 96.72%, 101.06%, 102.05%, and 99.03%, 100.18%, 100.28% in low, middle, and high concentrations, respectively. The average contents of schisandrin in serum and plasma were 11.063 2 and 12.883 7 ?g/mL, schisandrin B were 7.490 9 and 12.590 8 ?g/mL, respectively. Conclusion The main constitu-ents of Compound Wurenchun Capsules contained in plasma account for higher than the ones in serum.

OBJECTIVETo investigate the effects of lidocaine on apoptosis and proliferation of hyperoxia-exposed type Ⅱalveolar epithelial cells (AECⅡ) from premature rats.

METHODSPrimary cultured AEC Ⅱ isolated from premature rats were randomly divided into four groups: air, air+ lidocaine (20 μg/mL), 95%O2/5%CO2, and 95%O2/5%CO2+ lidocaine. The cells in each group were collected 24 hrs after culture in ordinary incubators (37℃,5%CO2). The proliferation and apoptosis of AEC Ⅱ were detected by flow cytometry. Protein levels of proliferating cell nuclear antigen (PCNA) were measured by Western blot.

RESULTSThe apoptosis rate of AECⅡincreased, the percentages of G2/M and S phase cells decreased and the protein levels of PCNA decreased significantly in the group exposed to 95%O2/5%CO2 compared with the group exposed to air (P<0.01). Lidocaine treatment decreased apoptosis rate of AECⅡ, increased percentage of G2/M and S phase cells, and increased protein levels of PCNA.

CONCLUSIONSHyperoxia can increase apoptosis and inhibit proliferation of AECⅡ in premature rats. Lidocaine may have protective effects against the AECⅡ injuries.

Animals ; Apoptosis ; Cell Cycle ; Cytoprotection ; Epithelial Cells ; drug effects ; pathology ; Female ; Hyperoxia ; pathology ; Lidocaine ; pharmacology ; Male ; Proliferating Cell Nuclear Antigen ; analysis ; Pulmonary Alveoli ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley

Background Basic fibroblast growth factor(bFGF)secreted by cornea after injury is important to cytothesis,collagen fibers reconstruction and axons recovery.However,the local bFGF is not enough for the reparation process.Objective This study aimed to observe the findings of corneal collagen and nerve recovery under the confocal microscopy through focusing(CMTF) in the eyes with intervene of exogenous recombinant bFGF(rbFGF) after excimer laser in situ keratomileusis(LASIK).Methods LASIK rabbit models were binocularly created in 34 clean New Zealand white rabbits.The tobramycin combined with dexamethasone were dropped after operation for 10 days in bilateral eyes.rbFGF was topically administered in the right eyes of rabbits from 1 day through 3 months after LASIK,and lubricant was used in the left eyes at the same way.The corneal collagen and nerve recovery,keratocyte and endothelial cell counting were observed with CMTF at the 1st week,2nd week,2nd month,3rd month and 5th month after LASIK.Results Total 19 rabbits were meted the request of LASIK models.The keratocyte densities in anterior stroma of both groups reached the lowest level at the 2nd week and the highest level at the 3rd month.Otherwise,haze changed on the contrary.No statistically significant differences were found in anterior stroma keratocyte densities,haze grade,grey value between rbFGF group and lubricant group at various time points after operation(P＞0.05).The nerve cord densities of both groups were increased gradually,and those under the epithelial basement membrane were more dominant.The nerve density of the anterior stroma of rbFGF group was significant higher than the lubricant one in the 2nd group(P=0.038).The considerably elevated the subepithelial nerve density value was also seen in rbFGF group compared with lubricant at 5 months after operation(Z=-2.060,P=0.039).No any corneal neovascularization occurred in both groups through experiment duration.The positive correlation was found between grey value with haze grade in rbFGF group(b=22.687,F=37.975,P=0.000) and lubricant group(b=20.410,F=18.516,P=0.000).However,haze grade was not significant correlated with stromal keratocyte density(rbFGF group:b=0.001,F=0.164,P=0.668;lubricant group:b=-0.002,F=1.896,P=0.178).Conclusion Exogenous bFGF can improve the recovery of corneal nerve and regeneration of keratocyte after LASIK.No evidence of bFGF promoting corneal neovascularization is found in this experiment.

OBJECTIVETo select optimal media and conditions for the tissue culture of Dendrobium lituiforum.

METHODThe basic media, light illumination condition, phytohormone, and natural aqueous extracts. were tested and compared. The chlorophyll contents and soluble protein contents were measured.

RESULTN6 medium was suitable for embryo germination and growth. 1/2 MS + NAA 0.5 mg x L(-1) cordd be used as the optimal protocorm multiplying media. Culture 20 days in darkness in advance, and then in light is useful to embryo germinate. 1/2 MS+ NAA 0-0.5 mg x L(-1) is beneficial to protocorm multiplication largely. N6 + 6-BA 2.0 mg x L(-1) + NAA 0.5 mg x L(-1) + 10% potato juice is useful to protocorm differentiation. Phytohormone and potato juice added to the media promoted chlorophyll content and souble protein content. N6 + NAA 0.5 mg x L(-1) + 10% banana juice is the best medium for plantlet rootage and strengthening.

CONCLUSIONConcentration of inorganic salt, nitrogen source and illumination condition are important to embryo germination and growth. The nitrogen source type has effect on the protocorm multiplication. The chlorophyll contents and souble protein contents may be index to indicate the growth condition of plantlets, which can help to select the optimal media.

Chlorophyll ; analysis ; Culture Media ; Dendrobium ; chemistry ; growth & development ; Light ; Plant Growth Regulators ; pharmacology ; Plant Proteins ; analysis ; Plants, Medicinal ; chemistry ; growth & development ; Tissue Culture Techniques

OBJECTIVETo investigate the in vivo inhibition of extract of Fructus lycii (FL) on the expressions of cathepsin B (Cat B) and cystatin C (Cys C) in high-fat diet and hydroquinone (HQ) induced model mice with age-related macular degeneration (AMD), and to explore the in vitro effects of lutein and zeaxanthin on hydrogen peroxide (H2O2,) induced expressions of matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase 2 (TIMP-2) on ARPE-19 cells.

METHODSFifty female 8-month-old C57BL/6 mice were recruited in this research. Ten mice fed with regular diet was taken as the age control group. The rest 40 mice were fed with high fat diet for 6 months, followed by adding HQ (0. 8%) in the drinking water for 3 consecutive months. Then the modeled mice were randomly divided into the model control group (n =10), the high (at the daily dose of 3.75 g/kg), middle (at the daily dose of 2.50 g/kg), and low dose (at the daily dose of 1.25 g/kg) FL groups, 10 in each group. The extract of FL at each dose was respectively administered to mice by gastrogavage for 3 successive months. By the end of the experiment, the mice were killed and their eyeballs were removed. The protein expressions of Cat B and Cys C were observed by immunohistochemical assay. The mRNA and protein expressions of Cat B and Cys C were detected by real-time PCR and Western blot respectively. The drug concentrations of H2O2, lutein, and zeaxanthin were screened and detected using the activity of cell proliferation. The protein expressions of MMP-2 and TIMP-2 were detected using Western blot.

RESULTSCompared with the age control group, the mRNA and protein expressions of Cat B and Cys C were significantly higher in the in vivo model control group (P <0.05, P <0.01). The mRNA expressions of Cat B and Cys C were weaker in the middle and high dose FL groups than in the model control group (P <0. 05, P <0. 01). In in vitro cells, lutein and zeaxanthin could down-regulate the protein expressions of MMP-2 and TIMP-2 in H202 induced ARPE-19 cells (P <0. 05, P <0. 01).

CONCLUSIONSExtract of FL could down-regulate the high protein expressions of Cat B and Cys C in high-fat diet and HQ induced model mice. Lutein and zeaxanthin could down-regulate the protein expressions of MMP-2 and TIMP-2 in H202 induced ARPE-19 cells.

Animals ; Cathepsin B ; metabolism ; Cystatin C ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hydrogen Peroxide ; Lutein ; pharmacology ; Macular Degeneration ; prevention & control ; Matrix Metalloproteinase 2 ; metabolism ; Mice ; Mice, Inbred C57BL ; Pigment Epithelium of Eye ; drug effects ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism ; Xanthophylls ; pharmacology ; Zeaxanthins