Objective To choose proper proton magnetic resonance spectroscopy(~1XH-MRS) parameters to fit for practical femoral marrow cavity and to produce short-timed,well-repeated and excellent ~1H-MRS images.Methods The tentative study of ~1H-MRS on the normal femoral bone marrow in 26 volunteers was performed with a 1.5 T MR after the informed consent.The single-voxel spectroscopy and stimulated echo acquisition mode were used for ~1H-MRS collection.~1H-MRS parameters for 12 volunteers were 128 acquisitions,1 cm?1 cm?1 cm volume of interest(VOI)size and repeatedly 2—3 times within the same location.~1H-MRS parameters for another:14 volunteers were different numbers of acquisition (128 and 256 times,respectively)and different VOI sizes(2 cm?2 cm?2 cm and 1 cm?1 cm?1 cm, respectively).Results For ~1H-MRS with 1 cm?1 cm?1 cm size of VOI and 128 times of acquisition with the full width haft max of water≤8—12 Hz,the base-line was steady and the signal-noise ratio was high up to 11.31.~1H-MRS was different in the different femoral locations showing the maximum peak sites at near 0.90 ppm(?10~(-6))or 1.65 ppm,but~1H-MRS within the same location was always same or similar with different VOI sizes(1 cm?1 cm?1 cm or 2 cm?2 cm?2 cm)or different numbers of acquisition(128 or 256 times).~1H-MRS acquisition time was not related with the size of VOI but with the numbers of acquisition.128 and 256 times of acquisition cost 199 s and 391 s,respectively.Conclusion With the technique of small size of VOI(1 cm?1 cm?1 cm)and decreased numbers of acquisition(128 times),it is propable to get well-repeated and excellent ~1H-MRS within less time.It is also more practical for clinics to achieve ~1H-MRS of the femoral marrow with the proper technique.

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

OBJECTIVETo study the relationship between the IFN-gamma producing cell specific for recipient (IFN-gamma-PCSR) in allogeneic stem cell transplantation (allo-HSCT) and acute graft versus host disease (aGVHD).

METHODSIn 37 consecutive HLA-identical sibling allo-HSCT pairs, peripheral blood mononuclear cells (PBMNC) from donors before allo-HSCT and recipients after allo-HSCT were taken as responder cells (RC), and PBMNC from recipients before allo-HSCT as allogeneic stimulator cells (allo-SC) in mixed lymphocyte reaction (MLR). IFN-gamma-PCSR in PBMNC were assayed using enzyme linked immunospot assay (ELISPOT).

RESULTSPretransplantation frequencies of IFN-gamma-PCSR in donor PBMNC were significantly higher in aGVHD group than in non-aGVHD group (P < 0.01) and IFN-gamma PCSRs (>or= 20/2 x 10(5)RC) were significantly associated with the occurrence of grade II-IV GVHD (P < 0.05). Compared with that before allo-HSCT, IFN-gamma PCSR in PBMNC of aGVHD patients was significantly increased (P < 0.05). When PBMNC from aGVHD patients reacted with donor PBMNC, the IFN-gamma PC was significantly lower than that with recipient PBMNC before allo-HSCT. Longitudinal analysis of IFN-gamma PCSR following allo-HSCT showed that compared with that in patients without aGVHD, the IFN-gamma PCSR were significantly higher in patients with that in aGVHD on day +14 (P < 0.01) and day +28 (P < 0.01), respectively. After immunosuppressive therapy for 7 days, IFN-gamma PC declined significantly (P < 0.05).

CONCLUSIONThe recipient-specific IFN-gamma PC is closely correlated with the allo-response during allo-HSCT and may be helpful for the prediction, diagnosis and monitoring of aGVHD.

Adolescent ; Adult ; Female ; Graft vs Host Disease ; immunology ; Hematopoietic Stem Cell Transplantation ; Humans ; Interferon-gamma ; biosynthesis ; immunology ; Lymphocyte Culture Test, Mixed ; Male ; Middle Aged ; T-Lymphocytes ; immunology ; metabolism ; Transplantation, Homologous ; immunology ; Young Adult

OBJECTIVETo detect, enrich and expand the cytokine secreting T lymphocytes after allogeneic PBMNCs stimulation.

METHODSThe novel cytokine secretion assay (CKSA) was applied to detect T lymphocytes secreting IFN-gamma at single cell level in human mixed lymphocytes reaction. IFN-gamma secreting T cells were enriched by means of magnetic sorting system and expanded with OKT(3), anti-CD(3)mAb and IL-2 combination. Antigen specificity of the expanded cells was confirmed using enzyme linked immunospot assay.

RESULTSA sizable proportion of IFN-gamma secreting T lymphocytes could be detected [(1.12 +/-0.13)% compared with (0.23 +/-0.07)%] and be further enriched to (67.3 +/-10.5)%, or (93.8 +/-22.1) fold. T lymphocytes could be expanded up to 600-fold within 21-28 days and the specific IFN-gamma response of expanded cells was confirmed with stimulation of the relevant allogeneic PBMNC, which was significantly higher than the irrelevant PBMNC control.

CONCLUSIONIt is feasible to detect significantly increased IFN-gamma secreting T lymphocytes after allogeneic PBMNCs stimulation based on the CKSA technique at single cell level and these cells can be efficiently enriched and expanded for further research.

Antibodies, Monoclonal ; pharmacology ; CD28 Antigens ; immunology ; Cell Proliferation ; Cells, Cultured ; Cytokines ; secretion ; Graft vs Host Disease ; immunology ; Humans ; Interferon-gamma ; secretion ; Interleukin-2 ; secretion ; Leukocytes, Mononuclear ; cytology ; immunology ; Lymphocyte Culture Test, Mixed ; Muromonab-CD3 ; pharmacology ; T-Lymphocytes ; cytology ; immunology

OBJECTIVETo investigate the inhibition effect of arsenic trioxide (AS(2)O(3)) on the growth of human MDS-RAEB cell line MUTZ-1 cells and to explore the possible cellular and molecular mechanisms.

METHODSThe apoptosis and differentiation of MUTZ-1 cells induced by AS(2)O(3) solution of different concentrations were studied with cell morphology, MTT, DNA fragmentation assay, RT-PCR, Nitroblue tetrazolium (NBT) reduction method and flow cytometry.

RESULT(1) Low concentration ofAS(2)O(3) (0.05 - 0.25 micromol/L) had no marked growth inhibition effect on MUTZ-1 cells; after 14 d treatment, it down-regulated the expression of positive cell differentiation antigens CD38, CD7, CD10, HLA-DR (P<0.05), but did not up-regulate the expression of CD11b (P>0.05). (2) After treatment with 1.0 - 20.0 micromol/L of AS(2)O(3), MUTZ-1 cells presented typical features of apoptosis with a dose dependent manner (r=-0.999, P<0.05). The expression of bcl-2 mRNA and the ration of bcl-2/bax were decreased after AS(2)O(3) treatment (P<0.05).

CONCLUSIONLow concentration of (2)O(3) may have partial differentiation inducement on MUTZ-1 cells. With a certain range of dose (1.0 - 20.0 micromol/L), (2)O(3) can induce apoptosis of MUTZ-1 cells. (2)O(3) can significantly down-regulate bcl-2 and it might be one of the mechanisms of (2)O(3) treatment.

Antineoplastic Agents ; pharmacology ; Arsenicals ; pharmacology ; Cell Division ; drug effects ; DNA ; analysis ; Dose-Response Relationship, Drug ; Flow Cytometry ; Humans ; Myelodysplastic Syndromes ; drug therapy ; pathology ; Oxides ; pharmacology ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; analysis ; bcl-2-Associated X Protein

When the glomerular mesangial cells of rats were cultured in vitro at high glucose concentration,the activity of extraeellular signal-regulated kinase(ERK),the expression of transforming growth factor-?_1(TGF-?_1)mRNA and the content of typeⅣcollagen in the supematant were higher than those at normal glucose concentration.These effects were inhibited by fluvastatin.The results showed that the activation of ERK signal transduction pathways appeared to play a role in the onset and progression of diabetic nephropathy. Furthermore,fluvastatin could protect the kidney by inhibiting ERK signal transduction pathway and TGF-?_1 expression.

Objective To explore the feasibility of evaluating the lung function by MSCT in emphysema.Methods The MSCT scan and pulmonary function tests(PFF)were respectively performed in 147 receptors within one week.They were randomly divided into 2 groups:group A(120 receptors), including normal,mild,moderate and severe abnormal pulmonary function based on the PFT,for comparing the correlation between pulmonary quantitative indexes of MSCT pulmonary function and PFT and settingup the primary grade criteria of abnormal pulmonary function in emphysema,group B(27 receptors)for evaluating the diagnostic accuracy in group A.The total lung was respectively scanned at the full inspiration and full expiration with MSCT.The pulmonary quantitative indexes of MSCT were measured with Siemens Pulmo pulmonary quantitative software.Results There was correlation between pulmonary quantitative indexes of MSCT and PFF.The Piex/in_(-910)showed best correlation with FEV_1%(r=-0.905,P

Sulfur fumigation, which is traditional method for preservation, pest control, insecticide and sterilization, has long been widely used in processing and storage and played a positive role of traditional Chinese medicine (TCM). As some businesses sided pursuit of profit, abused and repeated use of sulfur fumigation, have resulted in a large number of harmful residues, such as sulf dioxide (SO2) and harmful heavy metals, which brings a significant impact and danger on human health. This article summarizes the sulfur species and the sulfur fumigation methods and analyzes the harmful substances in TCM after sulfur fumigation, to provide a reference of the choice of species for the sulfur, the optimization of sulfur fumigation process and the standardized processing of TCM after sulfur fumigation.
Animals ; Drug Contamination ; Fumigation ; methods ; Humans ; Medicine, Chinese Traditional ; methods ; Safety ; Sulfur ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo analysis the changes of two chemical constituents, namely 2, 3-dihydro-3, 5- dihydroxy-6-methyl-4H-pyran-4-one (DDMP) and 5-hydryoxymethyl-furfural (5-HMF) produced in Radix Polygoni Multiflori after processing, with processing time, and to determine the contents of 5-HMF in samples of Radix Polygoni Multiflori and Radix Polygoni Multiflori preparata.

METHODAn HPLC method was applied with a Zobax SB-C18 (3.9 mm x 150 mm, 5 microm) column by a elution using methanol-water (10: 90) as the mobile phase. The detection was set at UV 280 nm.

RESULTThe contents of DDMP were increasing with the processing time until 24 hour, followed by a decrease until 60 hour process. The contents of 5-HMF were increasing gradually throughout the 60 hour steaming process. The contents of 5-HMF in 11 samples of Radix Polygoni Multiflori preparata were from 0.013% to 0.101%, and only one in 4 samples of Radix Polygoni Multiflori containing trace amount of 5-HMF.

CONCLUSIONThe chemical components in Radix Polygoni Multiflori were changed during the processing procedures. Therefore, the processing of Radix Polygoni Multiflori should be controlled and standardized.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Polygonaceae ; chemistry ; Reproducibility of Results

Objective To observe the effects of Postmenstrual Proliferative Prescription(mainly composed of Codonopsis Radix,Atractylodis Macrocephalae Rhizoma,Poria,Rehmanniae Radix Praeparata,Paeoniae Radix Alba,Angelicae Sinensis Radix,Chuanxiong Rhizoma,Cervi Cornu Degelatinatum,Corni Fructus,Cuscutae Semen,and Eucommiae Cortex)through replenishing qi and blood on the ovulation rate and pregnancy rate in patients with ovulatory dysfunction infertility caused by polycystic ovarian syndrome(PCOS)during ovulation-induction treatment,and to explore the therapeutic effects and possible therapeutic mechanism.Methods Sixty patients with ovulatory dysfunction infertility due to PCOS were randomly divided into a treatment group and a control group,with 30 patients in each group.The control group was given Clomifene Citrate Capsules to promote ovulation,and the treatment group was given Postmenstrual Proliferative Prescription on the basis of the ovulation-induction program of the control group starting from the fifth day of menstruation or progesterone withdrawal bleeding.The two groups were treated for one menstrual cycle as a course of treatment.The changes in the serum sex hormones of estradiol(E2),follicle-stimulating hormone(FSH),luteinizing hormone(LH),and progesterone(P)on the 2nd to 5th day of menstruation,as well as the changes in serum growth differentiation factor 9(GDF9)and bone morphogenetic protein 15(BMP15)levels on the 2nd to 5th day of menstruation and on the day of human chorionic gonadotropin(HCG)injection were observed in the two groups.Moreover,the ovulation rate,pregnancy rate and clinical efficacy of the patients in the two groups were analyzed.Results(1)No statistically significant differences in serum levels of sex hormones E2,FSH,LH and P on the 2nd to 5th day of menstruation were shown between the two groups of patients(P＞0.05).(2)On the 2nd to 5th day of menstruation and on the day of HCG injection,there were no significant differences in the serum GDF9 and BMP15 levels between the two groups(P＞0.05).(3)The ovulation rate and pregnancy rate in the treatment group were 93.33%(28/30)and 26.67%(8/30)respectively,which were significantly higher than 70.00%(21/30)and 13.33%(4/30)in the control group.And the differences tested by chi-square test were statistically significant between the two groups(P＜0.05).(4)The total effective rate of the treatment group was 93.33%(28/30),and that of the control group was 70.00%(21/30).The intergroup comparison(tested by rank sum test)showed that the therapeutic efficacy of the treatment group was significantly superior to that of the control group,and the difference was statistically significant(P＜0.05).Conclusion Postmenstrual Proliferative Prescription through replenishing qi and blood can improve the ovulation rate and pregnancy rate during ovulation-induction treatment in the patients with ovulatory dysfunction infertility due to PCOS.It is indicated that Postmenstrual Proliferative Prescription can enhance the quality of the oocytes and the potential of embryo implantation during the ovulation-induction treatment.