Objective To investigate the effects of cobalt chloride(COCl2)induced rat alveolar epithelial cell apoptosis and the significance of related apoptosis protein hypoxia inducible factor-lalpha(HIF-1 a)expression. Methods Alveolar epithelial cells were cultured in vitro and divided into four groups with 6 samples in each.Hypoxia 4 h group:added 500μmol/L COCl2 for 4 hours inducing alveolar epithelial cell hypoxia;hypoxia 24 h group:co-cultured with COCl2 for 24 hours;hypoxia 48 h group:co-cultured with CoCl2 for 48 hours;control group:no CoCl2 added.The apoptosis rates were detected by fluorescent microscope and flow cytometry.The apoptosis morphological changes of the cell were observed by transmission electron microscopy.The protein expression of HIF-la was detected bv Western Blot. Results (1)Fluorescent microscope results showed that apoptosis rate in hypoxia 4 h、24 h、48 h group[(5.83±0.76)%,(15.00±3.28)%and(51.50±3.00)%],were higher than control group[(1.50±0.50)%](P＜0.05).(2)Flow cytometry analysis reached the same results as with fluorescent microscope.(3)Typical morphological changes of apoptosis were observed in hypoxia 24 h group.(4)HIF-1α expression in hypoxia 4 h、24 h and 48 h groups[0.69±0.035,1.02±0.044 and 0.71±0.046],were higher than control group(0.21±0.026)(P＜0.01).(5)Positive correlations were found between relative amount of HIF-1αprotein and apoptosis rate by fluorescent microscope and flow cytometry analysis(r=0.484,P=0.016;r=0.713,P=0.009).Conclusions Hypoxia could induce apoptosis of alveolar epithelial cells,which may participate in the pathogenesis of hypoxia-induced lung injury.

Objective To explore the effects of different methods of antigen retrieval on the results of immunohistochemical staining of nuclear receptors. Methods Antigens were retrieved by using microwave oven, autoclave, enzyme and autoclave plus enzyme, respectively, in liver sections from DA.1U rats, followed by immunohistochemical staining. Results After antigen retrieval with autoclave, the positive staining intensity of nuclear receptors LXR-α, LXR-β, PPAR-γ and FXR in the liver sections from DA.1U rats was enhanced obviously, and the location of nuclear receptors was better by using this method than the others. Conclusion In the immunohistochemical staining of nuclear receptors, using autoclave to retrieve antigens is the best method.

OBJECTIVE: To explore the effect of different pressure on microcirculation of scar proliferation. METHODS: A total of 60 patients with scar proliferation after burning were collected, including 49 males and 11 females, with averagely aged 37 years. The patients were divided into low pressure (0.67-1.33 kPa), normal pressure (1.33-3.33 kPa) and high pressure (3.33-6.67 kPa) groups by envelope method. Pressure tension bandage was used at 1 week after wound healing and gradually increased, reached expected pressure at days 5-7 and sustained for 6 months. Then the changes of blood perfusion at the scar tissue were measured by laser Doppler perfusion imaging (LDPI). RESULTS: According to the intended processing analysis, all the 60 patients enter the result analysis. There were no significant differences of the scar tissue perfusion of the 3 groups before the therapy. After the therapy, the perfusion was decreased in the low pressure group, but there are no statistical changes compared to before the therapy (P > 0.05), while the perfusion was decreased in the normal pressure and high pressure groups (P < 0.05), which was lower in the high pressure group than the normal pressure group (P < 0.05). When the pressure increased to 3.33-6.67 kPa, the patients feel much more uncomfortable and the swelled signs appear, and 2 patients quit the experiment. CONCLUSIONS: The scar tissue perfusion is decreased obviously when adding the pressure to 3.33 kPa by using tension bandage. Using this method, the pressure should gradually increase to the maximum if patients can bear.

Objective To analyze the effect of bortezomib together with dexamethsone on the treatment of refractory multiple myeloma.Methods Bortezomib at 1.3 mg/m2 iv drop 1 d,4 d,8 d,11 d.Dexamethasonc 20 mg/d,1～4 d,8～11 d,17～20 d.21 days are used as a course.For bi-course dexamethasone was given by 20 mg/d,at 1～4 d.Results After 2～6 periods,3 cases achieved completely recovery(CR),4 cases are near to completely recovery(CR),3 cases got partially recovery(PR).Conclusion It is a good choice to treat refractory multiple myeloma by Bortezomib together with dexamethagone.

The paper aimed to find the optimal combination and evaluation of the interactions of antitumor effect of the curcumin (Cur) and adriamycin (ADM) in vitro. According to the factorial design and data characteristics, the parameter method combined with the response surface approach were used to analyze the pharmacodynamic interactions of in vitro antitumor effects of the combination of Cur and ADM at different dosages. The results showed that the dose-effect relationship of the combination with the ratio of ADM-Cur 1:3 showed significant differences in comparison with either used alone. The dose-effect curve was shift left in combination. The combination of adriamycin (ADM, 0.693-2.132 micromol L(-1)) and curcumin (Cur, 2.047-6.304 micromol L(-1)) with a fixed ratio (1:3) showed a synergism. With increasing doses of the combination, there is an additive effect. Computer simulation showed a trend of decreasing difference between the observed and expected effects with the dose increasing in Cur from 6.304 to 16.0 micromol L(-1) and ADM from 2.132 to 5.3 micromol L(-1). The response surface analysis showed the optimal combination to be Cur 18.50 micromol L(-1) and ADM 3.89 micromol L(-1) with a ratio of 5:1. This study suggests that the parameter method combined with the response surface analysis provides richer and more reasonable information, and is helpful for quantitative design of drug combination therapy and to describe the nature and degree of drug interaction.

AIM: To observe the effects of porous polyethylene ( Medpor) as a spacer graft in the reconstruction of large areas eyelid defect after the operation of malignant tumors of lower eyelids.
METHODS: Nineteen cases ( 19 eyes ) of malignant tumors of lower eyelid underwent the eyelid reconstruction were selected. Medpor lower eyelid inserts implantation were used to replace tarsal joint sliding conjunctival flap and pedicle flap, and repaired full -thickness lower eyelid defects then underwent eyelid reconstruction.
RESULTS: Appearance of eyelids and functional improvements were satisfactory with no stimulation on the eyeball and no effect on the visual function. Implants is with no absorption, shift, exclusion or infection and no tumor recurrence in all cases during the follow up for 6-36mo.
CONCLUSION: Medpor lower eyelid inserts implantation can instead tarsal plate for the reconstruction of medium to large areas lower eyelid defect, which is easy performing with rare complications. It is an ideal alternatives of tarsal plate.

Objective:To analyze the protective effects of adoptively transferring different patrilineal lymphocytes and their Exosomes ( Exo) on fetation of mice with pregnancy loss comparatively.Methods: The peripheral blood mononuclear cells ( PBMC) from healthy men and the splenocytes from BALB/c and DBA/2 male mice were induced in vitro ,and their Exo were isolated through sucrose gradient ultra-centrifugation combined with ultrafiltration.The mice of CBA/J (♀) mated with BALB/c (♂) were enrolled as control group of normal pregnancy ,and the CBA/J (♀) mated with DBA/2 (♂) as URSA of pregnancy loss experimental animal model.The mice in URSA group were randomly divided into each group with treatment through adoptively transferring , which were injected intravenously or subcutaneously with splenocytes or splenocytes -derived Exo from mated DBA/2,unmated DBA/2 or unrelated BALB/c,also PBMC-derived Exo from men,respectively.And then,the placenta volumes,rates of fetal absorption and pregnancy loss were calculated to observe the fetation of embryos.Results:Compared with the group of normal pregnancy ,the placenta volumes from URSA group decreased greatly ,and rates of fetal absorption and pregnancy loss elevated greatly ( all P<0.000 5 ).After transferring different sources of cells and their Exo through different injection ,the placenta volumes resumed to the level of normal pregnancy ,and the rates of fetal absorption and pregnancy loss decreased significantly ( all P<0.000 5 ).No differences were observed after treatment through injecting intravenously or subcutaneously ( all P>0.05 ).After transferring the Exo derived from either male mice or healthy men,the level of decreased fetal absorption rates were more than that in cellular-therapy groups ( all P<0.05 ).After transferring the Exo derived from men ,the level of decreased pregnancy loss rates were more than that in cellular -therapy groups and mice splenocytes-derived Exo group ( all P<0.05 ).Conclusion:Adoptively transferring patrilineal T lymphocytes and their Exo can greatly improve the fetation.Exo should become a non-cellular bio-remedy,which is expected to replace traditional immunotherapy of adoptively transferring lymphocytes.

Objective To explore the diagnostic value of two-dimensional speckle tracking technology in the evaluation of right ventricular systolic function in patients with pulmonary disease .Methods Thirty patients with pulmonary heart disease were divided into two groups:group of compensated pulmonary heart disease(compensated group ) (n =15),group of decompensated pulmonary heart disease (decompensated group)( n =15).30 healthy subjects were enrolled in control group .The displacement of the tricuspid annulus at the midpoint(TADmid),the displacement of the tricuspid annulus at the free wall (TADfre) and the displacement of the tricuspid annulus at the septum (TADsep) were acquired,and simultaneous real-time three-dimensional ultrasound detection of right ventricular ejection fraction (RVEF) were taken.The correlation of TADmid with RVEF and pulmonary artery systolic pressure ( PASP) were analyzed.Results TADmid of the healthy control group,the compensated group and the decompensated group were(17.1 ±3.9)mm, (13.6 ±2.6)mm,and(9.5 ±3.2)mm respectively.TADfre were(21.1 ±3.0)mm,(17.6 ±4.2)mm,and (11.5 ±3.8) mm respectively.TADsep were(12.0 ±2.5) mm,(9.7 ±3.3) mm,and(7.4 ±2.7) mm respectively.RVEF were(56.3 ±8.2)%,(39.6 ±6.4)%,and(28.1 ±5.9)% respectively.PASP were (20.6 ±2.6) mm Hg (1 mm Hg =0.133 kPa), (63.3 ±5.6) mm Hg, and (82.5 ±11.2)mm Hg respectively.There were significant differences of TADmid , RVEF, and PASP among the 3 groups ( F =8.581,7.816,9.300,6.507,10.235, all P <0.05).TADfre, TADmid, TADsep and RVEF were all decreased in the compensated group comparing to the healthy control group ,while PASP was increased.The decrease of TADmid was the most significant ,while that of TADfre was the slightest .There were significant differences of TADfre,TADmid,TADsep,RVEF and PASP between the 2 groups(t=2.703,2.536,2.379, 2.817,3.026,all P<0.05).TADfre,TADmid,TADsep and RVEF of decompensated group reduced more significantly than the compensated group , while PASP was increased significantly .There were significant differences of TADfre,TADmid,TADsep,RVEF and PASP between the 2 groups(t=2.519,2.493,2.236, 2.621,2.985,all P<0.05);TADfre and TADmid were decreased more apparently in decompensated group than that in compensated group ,and so were TADsep and RVEF,while PASP were increased.There were significant differences of TADfre,TADmid,TADsep,RVEF and PASP between the 2 groups (t =1.947, 2.680,2.016,2.653,2.893,all P<0.05).There was significant positive correlation between TADmid and RVEF measured by real-time three-dimensional ultrasound (r =0.904,P <0.01 ).There was significant negative correlation between TADmid and PASP (r=-0.686,P<0.01).The cut-off point value of TADmid measured by speckle tracking technology for evaluation of RVEF <45% and<30% were 13.65 mm and 9.80 mm,respectively.The sensitivity were 94.4%and 90.0%respectively,and the specificity were 78.6%and 90.0%respectively.Conclusions TADmid is hardly affected by the external factors ,and it can better reflect the changes of right ventricular systolic function for patients with pulmonary heart disease .TADmid is positively correlated with RVEF measured by real-time three-dimensional ultrasound and negatively correlated with PASP.Further more,the correlations is significant.The three parameters can authenticate mutually ,and the combination of them can evaluate the right ventricular systolic function in patients with pulmonary heart disease precisely.

The powder X-ray diffraction Fourier fingerprint pattern technique was used to develop a new quantitation method for the analysis of andrographolide and dehydroandrographolide. And the high performance liquid chromatography method was used to evaluate the quantity of andrographolide and dehydroandrographolide. The relationship of diffraction peak intensity and content of andrographolide and dehydroandrographolide was investigated. The powder X-ray diffraction Fourier fingerprint pattern analysis technique can be used to evaluate the quantity of andrographolide and dehydroandrographolide in the herb simultaneously.
Andrographis ; chemistry ; Chromatography, High Pressure Liquid ; Diterpenes ; analysis ; Drugs, Chinese Herbal ; analysis ; Fourier Analysis ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Powders ; X-Ray Diffraction ; methods

OBJECTIVETo explore the association of the expressions of human leukocyte antigen (HLA)-DR4, peptidyl arginine deiminase type4(PAD4), and signal transducer and activator of transcription 4 (STAT4) in the peripheral blood with the disease activity in patients with rheumatoid arthritis (RA).

METHODSTwenty-four RA patients in active stage (DAS28 score>or=2.6) and 14 RA patients in remission stage (DAS28 score<2.6) were enrolled in this study, with 12 healthy volunteers as the control. The QuantiGene Plex method was used to measure the expression level of HLA-DR4, PAD4, and STAT4 mRNA, and the relationship between the expressions of these genes and the DAS28 score, levels of anti-cyclic citrullinated peptide antibody (anti-CCP antibody) and rheumatoid factor (RF) was analyzed.

RESULTSThe expressions of HLA-DR4, PAD4, and STAT4 were significantly higher in RA patients than in the healthy controls (P<0.05). The level of HLA-DR4 mRNA in the two RA groups showed no significant difference, but was significantly higher than that in the healthy controls. HLA-DR4 expression was not found to correlated to DAS28 score, anti-CCP antibody level or RF in the RA patients. The expressions of PAD4 and STAT4 were significantly different between the two RA groups (P<0.05). In the RA patients, PAD4 mRNA expression was positively correlated to DAS28 and anti-CCP antibody level (P<0.05), and STAT4 expression showed positive correlations to DAS28 and RF levels (P<0.05).

CONCLUSIONHLA-DR4, PAD4 and STAT4 are overexpressed in RA patients and may be involved in the pathogenesis of RA. The expressions of PAD4 and STAT4, but not HLA-DR4, are closely related to the disease activity of RA. Detection of peripheral blood PAD4 and STAT4 expressions can be helpful for evaluating the disease activity of RA.

Adult ; Arthritis, Rheumatoid ; blood ; Female ; HLA-DR4 Antigen ; blood ; genetics ; metabolism ; Humans ; Hydrolases ; blood ; genetics ; metabolism ; Male ; Middle Aged ; Protein-Arginine Deiminases ; RNA, Messenger ; genetics ; metabolism ; STAT4 Transcription Factor ; blood ; genetics ; metabolism