BADH-CMO double gene,CMO gene and DREB1A gene were transformed respectively to embryonic callus of Kentucky bluegrass by particle bombardment. Then the embryonic callas of kentucky bluegrass were put in meclium for subculture which is mixed with 100mg/L hygromycin and the meclium for plantlet regeneration which mixell with 50mg/L hy gromycin about one month. Thus,the hygromycin-selectecl plants were obtained and were transplantecl into tlowerpots. The results of the PCR and Southern blot analysis indicated that the DREB1A gene,CMO gene and BADH-CMO double-gene were integrated into the genomic DNA of Kentucky bluegrass.

Objective To explore the effects of stimulating vagus nerve with pulse current on peripheral blood CD4+T lymphocyte of rats with collagen induced arthritis and its mechanism.Methods To duplicate model rats of experimental arthritis(EA)by intradermal injection of Ⅱtype collagen,divide the rats into 2 groups:vagus nerve stimulation(VNS)group and sham operated group.Rats in VNS group were stimulated at the left cervical vagus nerves for 30 minutes a day with constant square wave,pulse current with intra train of 16 Hz,pulse duration of 1.0 ms,train duration of 10 s,interstimulus interval of 1.5 min and intensities of 3.0 mA.Then flow cytometry and immunofluorescence methods were used to detect the activation of CD4+T lymphocytes(expressing CD71)and the expression of nicotinic acetylcholine receptors alpha 7(nAChR?7)and choline acetyltransferase(ChAT)in peripheral blood CD4+T lymphocytes.Results In VNS group,the expression of nAChR?7 and ChAT were significantly raised in CD4+ T cells at 1st weekend(Pa

To study effect of vitrification state of protective solutions on recovery of red blood cells after lyophilization, four protective solutions composed of isotonic buffers containing 7% DMSO (v/v) and 20%, 30%, 40% or 50% polyvinylpyrrolidone (PVP) (w/v) were adopted. Vitrification state of protective solutions was examined first when white ice crystal appeared in any protective solution during freezing or thawing, if the used solution was not a vitrification solution. Red blood cells were lyophilized in MINILYO45 freeze-dryer after washing, mixing with protective solutions and prefreezing. After lyophilization, the samples were quickly rehydrated by 37 degrees C rehydration solution. The results showed that in vitrification and devitrification experiments, white ice crystal appeared in solution of 20% PVP + 7% DMSO and 30% PVP + 7% DMSO during freezing and thawing; vitrification appeared in solution of 40% PVP + 7% DMSO during freezing, but devitrification appeared during thawing; vitrification appeared in solution of 50% PVP + 7% DMSO during freezing and thawing. After rehydration, the recoveries of red blood cells and hemoglobin in 40% PVP + 7% DMSO group were (81.36 +/- 14.94)% and (77.54 +/- 12.86)%, which were significantly higher than that in 20% PVP + 7% DMSO, 30% PVP + 7% DMSO and 50% PVP + 7% DMSO groups (P < 0.01). The concentration of free hemoglobin in 40% PVP + 7% DMSO group was also significantly lower than that in other three groups (P < 0.01). With increase of PVP concentration in protective solutions, vitrification state and protective effect of these solutions also increased; when concentration of PVP in protective solution was 40% though it was not a vitrification solution, the effect of lyophilization was the best; but when concentration of PVP further increased to 50%, though it was a vitrification solution, the effect decreased. It is concluded that excessive vitrification state could not benefit lyophilization of red blood cells.
Cryoprotective Agents ; pharmacology ; Dimethyl Sulfoxide ; pharmacology ; Dose-Response Relationship, Drug ; Erythrocytes ; cytology ; drug effects ; ultrastructure ; Freeze Drying ; methods ; Humans ; Microscopy, Electron ; Povidone ; pharmacology

To investigate the mechanism of agarwood formation in Aquilaria sinensis induced by Lasiodiplodia theobromae, the fermentation liquor of L. theobromae was analyzed qualitatively and quantitatively by gas chromatography-mass spectrometry (GC-MS). JAs were detected in the fermentation liquor. The effect of the fermentation liquor on the abundance of sesquiterpenes in the callus of A. sinensis was analyzed by solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). And the fermentation liquor stimulated alpha-guaiene, alpha-humulene and delta-guaiene biosynthesis in calli. It was inferred that L. theobromae produced JAs, which resulted in a significant increase of sesquiterpenes in A. sinensis.
Ascomycota ; physiology ; Fermentation ; Sesquiterpenes ; metabolism ; Thymelaeaceae ; metabolism ; microbiology

OBJECTIVETo explore the effect of Yiqi Huoxue recipe (YHR), a Chinese herbal medicine for supplementing Qi, activating blood circulation to remove stasis, on vascular extracellular matrix (ECM) remodeling and its molecular mechanism.

METHODSVascular smooth muscle cell (VSMC) proliferation activity and collagen turnover rate were detected by 3H-TdR test and hydroxyproline amount determined by 3H-Pro incorporation. Expression activity of MMP-2 and osteopontin genes was detected by Northern blotting and MMP-2 zymography analysis.

RESULTSYHR could markedly inhibit VSMC collagen synthesis stimulated by blasic fibroblast growth factor (bFGF) and lower the collagen turnover rate induced by vascular de-endothelialization. The expression level of MMP-2 and osteopontin genes was down-regulated by YHR in cultured VSMC and vascular wall with endothelial injury, and VSMC proliferation was inhibited by the serum obtained from YHR treated rats. Removing protein from the drug serum made no change on the effect of YHR to VSMC.

CONCLUSIONYHR could inhibit and/or retard ECM remodeling through regulating the expression of MMP-2 and osteopontin genes and lowering the collagen turnover rate.

Animals ; Aorta ; cytology ; Cell Division ; drug effects ; Cells, Cultured ; Collagen ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Matrix ; genetics ; metabolism ; Fibroblast Growth Factor 2 ; metabolism ; Gene Expression ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Osteopontin ; Rats ; Rats, Sprague-Dawley ; Sialoglycoproteins ; genetics ; metabolism

Objective To determine the expression of membrane-bound B lymphocyte stimulator (BLyS) protein and its mRNA in vitro of peripheral blood mononuclear cells (PBMCs) from individuals with systemic lupus erythematosus (SLE),and to investigate the role of interferon-?(IFN-?) on the expression of BLyS.Methods PBMCs were obtained from 25 SLE patients (mean age of 31+14) and 20 healthy volunteers (mean age of 28?10).They were randomized into IFN-?(5 ng/ml) group and control group.PBMCs were col- lected at 0,6,12 and 24 h for BLyS mRNA assessment using semi-quantitative reverse transcription-PCR (RT-PCR).PBMCs were also collected at 72 h for membrane-bound BLyS protein detection using flow cy- tometry (FACS) and direct immunofluorescence.Results①The expression of BLyS mRNA and membrane- bound protein in PBMCs was significantly higher in individuals with SLE compared with healthy controls (P＜0.05);②IFN-?enhanced BLyS mRNA expression in PBMCs in both healthy controls and SLE patients,with the greatest effect at 6 h (stimulated vs unstimulated,0.42?0.19 vs 0.25?0.14,P＜0.01;0.59?0.28 vs 0.44?0.21,P＜0.01 );③IFN-?also increased the expression of membrane-bound BLyS protein in both healthy con- trols and individuals with SLE (FACs,mean fluorescence intensity,4.5+3.0 vs 3.7~2.6,P

Objective To explore the characteristics of human plague using a sero-epidemiologic method in the source of the three rivers area in Qinghai for possible plague control strategies. Methods Investigate human plague sero-epidemiologically in the source of 4 counties in the three rivers area in Qinghai. The human serum would be tested to confirm the sew-positive rate for plague F1 antibody using indirect hemagglutination assay(IHA). Results A total of 2508 local participants were tested in 4 counties, the overall plague sero-positive rate was 2.31%(58/2508). This represents a statistically significant difference with 4 counties(X2=19.30,P<0.01). The sew-positive rate for males and females were 2.54% (32/1261) and 2.09% (26/1247), respectively. There were no statistically significant differences between males and females(X2= 0.65,P 0.05). The sero-positive rate in herdsman, cadre, Tibetan, Hart nationalities were 3.54% (44/1243), 6.47% (11 / 170), 2.40% (56/2335) and 1.47% (2/136), respectively. The sero- positive rate increased with age. The highest titre for human plague serum antibody was 1 : 640. Conclusion There were occult infections of plague in the population on source of three rivers area in Qinghai. Sero-epidemiologic data revealed that the human plague sero-positive rate was closely correlated with the local animal plague.

Objective To investigate the expression of vascular endothelial growth factor(VEGF)in synovium of rats with adjuvant arthritis and the relationship between the histopathologic score and the expression of VEGF.Methods Adjuvant arthritis was established in Wistar rats by inoculating complete Freund's adjuvant(CFA). We calculated the arthropathologic score and the expression of VEGF mRNA and protein at different stages after CFA inoculation.Results In model group the arthropathologic score and expression of VEGF protein in synovium increased significantly all the time (P

BACKGROUNDThere is no consensus regarding the performance for endorectal ultrasonography (ERUS) at every stage of rectal cancer. Thus, the purpose of our study was to further assess the value of ERUS in the preoperative staging of rectal cancer.

METHODSA retrospective study was performed with 44 consecutive patients (mean age: (63.3 ± 10.2) years) who underwent surgical treatment for endorectal carcinoma and were preoperatively evaluated using Biplane ERUS between September 2008 and December 2010. We compared the ERUS staging with the pathological findings based on surgical specimens.

RESULTSERUS staging agreed with the histologic staging in 39 of the 44 (88.6%) patients: the agreement on the depth of transmural invasion was good (κ = 0.73; 95%CI: 0.60 - 0.86, P = 0.000). The detection sensitivities of rectal cancer with ERUS were as follows: T1 85.7%, T2 87.5%, T3 88.9%, and T4 100.0% with specificity values of T1 97.3%, T2 92.9%, T3 96.2%, and T4 97.6%. ERUS correctly staged patients with T1 95.5%, T2 90.9%, T3 70.5%, and T4 97.7%. The positive predictive value of ERUS was lowest for T4 (75%), but highest for T3 (94.1%) followed by T2 (87.5%) and T1 (85.7%); the negative predictive values of ERUS from high to low were ordered as T4 (100%), T1 (97.3%), T2 (92.9%), and T3 (92.6%). The percentage of total over-staged cases was 4.5% and the under-staged cases was 6.8%. The extent of perirectal lymph node metastases was determined with a sensitivity of 68.4% (13/19), specificity of 80.0% (20/25), and diagnostic accuracy of 75.0% (33/44).

CONCLUSIONBiplane ERUS has a high diagnostic accuracy for tumoral invasion of the rectal wall at every T stage, but relatively low diagnostic accuracy for lymph node metastases.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Rectal Neoplasms ; diagnostic imaging ; pathology ; Retrospective Studies ; Ultrasonography

OBJECTIVETo study the feasibility of close and three-dimensional reconstruction of maxillary defects combination with pedicled buccal fat pad graft, temporalis myofascial flap and titanium mesh.

METHODSAmong 11 patients with maxillary tumors, 9 were malignant and 2 were benign. 8 cases underwent subtotal maxillectomy and 3 cases underwent total maxillectomy. Maxillary defects were closely reconstructed combination with pedicled buccal fat pad graft (BFP), temporalis myofascial flap (TMF) and titanium mesh to restore the profile of maxilla.

RESULTSAll flaps were alive with satisfactory function and aesthetic appearance. The uncovered BFP and the facial muscle surface of TMF epithelialized within 4-6 weeks. Their degree of mouth opening ranged from 2.5 to 4.0 cm and without nasal foods return. All patients had a good voice quality.

CONCLUSIONMaking use of BFP, TMF and titanium mesh can restore the good contour and function of maxillary. It is an effective method of reconstruction of maxillary defects, and is easy to manipulate. It is worthy to popularization.

Adipose Tissue ; Female ; Humans ; Male ; Maxilla ; Maxillary Neoplasms ; Middle Aged ; Prostheses and Implants ; Reconstructive Surgical Procedures ; Surgical Flaps ; Titanium