Objective To clone human asparagine synthetase(ASNS)gene,express the MS2- ASNS fusion protein through gene engineering and use the purified target protein to immune BALB/C mice, prepare and identify the monoclonal antibody(McAb),which forms the base for studying mechanism of L- asparaginase used as salvage regimen in midline NK/T cell lymphoma nasal type.Methods ASNS gene fragment was amplified by RT-PCR from HepG2 cell line and constructed into prokaryocytic expression vector.Fusion-protein of MS2-ASNS was expressed and used for immunizing BALB/C mice to prepare McAbs against ASNS.Identified the McAb and detected the expression of ASNS in tumor.Results Part of the ASNS reading frame(NCBI,M27396:179-1 420 bp)was cloned and product length of RT-PCR was 1 263 bp.Molecular weight of MS2-ASNS was about 54 700 Da.Two strains of hybridoma secreting ASNS McAbs were obtained.The subtype of the ASNS McAb was IgG2a.Western-blot showed that the McAbs could specifically react with MS2-ASNS fusion protein and ASNS protein in tumor cell lines.ASNS expression was detected by immunocytochemistry and Immunohistochemisrry.Conclusion We have cloned human ASNS gene,obtained the anti-ASNS McAb and examined the expression of ASNS in tumor.

Objective To investigate the curative effects of curcumin(CUR)or dexamethasone (DXM)on ischemia-reperfusion injury(IRI)of rat lung grafts.Methods Male SD rats were randomly divided 4 groups:CUR group(CUR was administered intraperitoneally to both donors and recipients at 3 h prior to operation);DXM group(DXM was administered intraperitoneally at 30 min prior to operation);vehicle group(Animals were injected with the DMSO to both donors and recipients at 3 h prior to operation);sham group(Time-matched control animals underwent the same surgery,except that no graft was implanted).Six animals were sacrificed at different reperfusion periods of 2 h and 24 h,respectively.Oxygenation indexes(PO_2/FiO_2),lung injury scores,wet/dry ratio(W/D)and myeloperoxidase(MPO)activity in the transplanted lung were measured.Malondialdehyde(MDA), total anti-oxidative capacity(TAOC),tumor necrosis factor(TNF)-?and interleukin(IL)-6 in the transplanted lung and serum were determined.Results The levels of LPV PO_2/FiO_2 were significant- ly higher in the CUR and DXM groups than in the vehicle groups both 2 and 24 h after reperfusion,re- spectively(P

Background and purpose:Most breast cancer cell lines were derived from malignant pleural fluid or other tissues that metastasized from breast carcinoma.Breast carcinogenesis is under represented due to the significant distinction of biological characteristics between cancer cell lines and primary breast cancers.Few primary breast tumors spontaneously developed into immortal cell lines.Overexpression of human telomerase reverse transcriptase(hTERT) in normal epithelial cells led to immortalization.We aimed to establish immortalized cultures derived from primary breast cancer by introducing hTERT and determine the gene profile of hTERT-transfected cultures.Methods:hTERT was introduced into breast tumor cultures with a puromycin-resistant retroviral construct,pBABE-hTERT-puro.Real-Time RT-PCR was used to analyze the expression level of hTERT after transfection.The upregulated gene profile was determined with microarray techniques.Results:hTERT was stably over expressed,at least 1 400-fold in hTERT-transfected cells compared to pBABE-puro transfected cultures.These ectopic over-expressions led to immortalization of transfected cultures,which could undergo at least 40-50 passages.Microarray analysis further confirmed the over-expression of hTERT in transfected cultures and demonstrated that 22 genes were up-regulated due to over-expression of hTERT.No down-regulated genes were observed cross all cultures.Conclusions:Over-expression of hTERT led to immortalization of breast tumor primary cultures and up-regulated certain genes.

Background and purpose:Overexpression of human telomerase reverse transcriptase(hTERT)plays a critical role in the process of immortalization of tumor cells.This study aims to investigate siRNA's effects on endogenous hTERT mRNA level and growth of breast cancer cells by targeting hTERT with its specific siRNA.The ultimate goal was to further elaborate the possible mechanism of immortalization in breast cancer cells.Methods:Real-Time RT-PCR was used to determine the hTERT transcripts in various breast cancer cells.hTERT siRNA was transfected into MCF7 cells with lipofectamine 2000.Cell growth was illustrated by growth curve,and cells apoptosis percentage was measured with Flow Cytometry(FACS).The relative expression levels of immortalization-related genes were evaluated with Real-Time RT-PCR.Results:Overexpressions of hTERT were demonstrated in all tested breast cancer cell lines.The inhibitory effects of hTERT siRNA were shown on both hTERT mRNA levels and cell growth from day 4 post transfection.Apoptosis was induced by hTERT siRNA as well.Certain immortalization-related genes were reduced by more than 50%,such as RAC1,PCYT2,FDFT1 and ATP5G2.Conclusions:hTERT siRNA specifically inhibited hTERT at mRNA level as well as cell growth.The apoptosis and downregulation of immortalization-related genes due to hTERT siRNA was demonstrated.

Background and purpose:The effects of triiodothyronine(T3)on breast cancer remain unclear.The aim of this study was to investigate the expression and correlation of estrogen receptor-?(ER-?)and thyroid hormone receptor-?(TR-?)in breast cancer cell lines,and to determine the changes of relevant receptors expression and its possible mechanism due to T3 treatment.Methods:Real-Time PCR was used to analyze the expressions of ER-? and TR-? at mRNA level in seven breast cancer cell lines.The expression level of retinoid X receptor-?(RXR-?)and these two receptors after T3 treatment were determined as well.The gene profile was further determined in SK-BR-3 cells treated with T3.Results:The expressions of ER-? and TR-? were quite low in SK-BR-3,MDA-MB-231 and MDA-MB-435 cells,but high in MCF7,BT20,BT474 and T47D cells.ER-?,TR-? and RXR-? were upregulated due to T3 treatment in ER-?(-)cells,but not in ER-?(+)cells.In SK-BR-3 cells,T3 treatment led to up-regulation of E2F1,TGF-?1R,TGF-?2R and hTERT,but down-regulation of TGF-?1 and p21.Conclusions:The expressions of ER-? and TR-? are correlated in breast cancer cells;the effect of T3 treatment on hormone receptors is associated with endogenous ER-? level.

The aim of this study is to develop the Tripterygium glycosides nanoemulsion gels and investigate its pharmacodynamics. Oleic acid was used as oil phase, polyoxyethylene castor oil as surfaetant, and 1,2-propanediol as cosurfactant to screen the formula of Tripterygium glycoside nanoemulsion using the pseudo-temary phase diagrams. Then the nanoemulsion gels was prepared. The ICR mouse ears were sensitazated by 7% DNCB, and then were excited by 0.3% DNCB to stimulate the model of mouse chronic dermatitis and eczema. The concentrations of IFN-γ, IL-4 and IL-8 in mouse blood were determined by ELISA. The results showed that Tripterygium glycosides nanoemulsion gels could significantly inhibit the swelling of mouse ears(P < 0.01) and ameliorate the edama and erythema of model mouse ears skin. Also it could significantly decrease the expression of IFN-γ and IL-4 in model mouse blood. Tripterygium glycosides nanoemulsion gels had a good therapeutic effect on mouse model of dermatitis and eczema. It was expected to provide a new and long-acting exterernal preparation for the treatment of dermatitis and eczema.
Animals ; Chemistry, Pharmaceutical ; instrumentation ; methods ; Dermatitis ; drug therapy ; immunology ; Drug Carriers ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Emulsions ; chemistry ; Female ; Glycosides ; chemistry ; pharmacokinetics ; Humans ; Interleukin-4 ; immunology ; Interleukin-8 ; immunology ; Mice ; Mice, Inbred ICR ; Nanoparticles ; chemistry ; Tripterygium ; chemistry

Group A,the differences among the three groups were statistically significant(P0.05). TCR,CCR,DAP,OD,BRC in Groups B and C showed trends of increasing,the differences in terms of the contents of BMP-RⅠamong the 3 phases were statistically significant(P0.05);the ER in Group B and C showed a trend of decreasing,thee difference between 4- and 8-week and 4- and 12-week were signifieantly dif- ferent(P0.05).Conclusion After application of glucocorticoid for a short term,pathological changes maintained and showed trends of inereasing in early stage.HBO can reverse these changes.The outcome of 3-course HBO therapy is better than that of 1-course therapy.

Objective The incidence of complications after esophageal cancer surgery is relatively high.The purpose of this paper was to explore the feasibility of combined thoraco-laparoscopy surgical treatment for senile esophageal cancer(over 70 years old).Methods A total of 526 esophageal cancer cases were retrospectively analyzed in this study.The operative procedures was esophageal carcinoma resection surgery and thoracic and abdominal lymphadenectomy which were operated through video-assisted by thoracoscope and laparoscopic From January 2010 to June 2014 in the Union Hospital of Fujian Medical University,divided into aged groups(≥70 years old,n =132) and non-aged group(＜ 70 years old,n =394).Statistical analysis was carried on the rate of two groups of preoperative risk factors(hypertension,diabetes,cardiac insufficiency,pulmonary insufficiency,cerebral infarction),surgical trauma and the number of lymph node dissection (laryngeal recurrent nerve injury,operation time,intraoperative blood loss),postoperative complications(cardiac complications,chylothorax,pulmonary complications,anastomotic fistula,poor healing of the incision,intrathoracic stomach atony),postoperative recovery and early postoperative mortality.Results Preoperative risk factors:the prevalence of the aged group with hypertension,cerebral infarction,cardiac insufficiency,and pulmonary insufficiency was significantly higher than the non-aged group (P ＜ 0.05) but diabetes (P ＞ 0.05).Surgical trauma and the number of lymph node dissection:no significant difference (P ＞ 0.05).Postoperative complications:the aged group was higher in the incidence of pulmonary complications and cardiac complications than the other (P ＜ 0.05) ; but the chylothorax,anastomotic fistula and poor healing of incision,intrathoracic stomach atony in the two groups seemed no significant difference(P ＞ 0.05).Furthermore,pulmonary complications were highest in the all complications,significantly higher than the others in both groups.Postoperative recovery:significant difference existed in postoperative hospitalization days,the aged group was obviously longer than the non-age groups.Early postoperative mortality rates in the two groups had no significant difference.Conclusion The aged patiences had a higher prevalence in the preoperative risk factors and so was the cardiac complications as well as pulmonary complications among the postoperative complications.However the combined thoraco-laparoscopy in the surgical treatment of elderly patients with esophageal cancer is safe and feasible.

Objective　To evaluate the changes and clinical s ignificances of serum growth hormore (GH) and GH binding protein (GHBP) in patie nts with obstructive jaundice (OJ). Methods　Serum levels of GH, GHBP, insulin-like growth factor-Ⅰ (IGF-Ⅰ), endotoxin, prealbumin concentr ation and liver function in 60 male in patients with OJ were detected 1 d preope ratively and 1, 3, 7 d postoperatively. And 40 male patients with simple gallbla dder stone hospitalized in the same period were served as control. Resu lts　The levels of GH, GHBP and IGF-Ⅰ were significantly lower in pati ents with obstructive jaundice than in control group(P<0.01), which was corr elated with the levels of bilirubin, endotoxin and prealbumin (P<0.01). Conclusion　The obviously downregulation of GH/GHBP axis in patients with OJ probably results from the inbibition of expression of GH receptor due t o nutritional dysfunction and endotoxemia, and further aggravate the nutritional dysfuction and endoxemia, which may be one of the important factors in the path ophysiological changes of OJ.

Objective To analyze the high resolution CT(HRCT)appearance and its clinical significance in patients with post-traumatic visual loss.Design Retrospective study.Participants 150 patients with post-traumatic visual loss.Methods The optic canal HRCT of 150 patients admitted from March to June of 2007 by Department of Emergency for post-traumatic visual loss were ana- lyzed retrospectively.Main Outcome Measures Pedilection sites and their incidence rates of optic canal and orbital apex region. Results Optic canal fractures were found in 78 patients,manifesting as rupture and displacement of optic canal wall,accompanied with or without adjacent structure fractures.(1)Among 88 sides of fractures,22 sides presented with single fracture,and 66 sides with multi- ple fractures,revolving several optic canal walls.(2)156 sites of fractures were displayed in 78 patients,and the most frequent types were displacement fractures(47.4%).(3)Accompanied adjacent structure fractures were found in 73 cases of the 78 patients,while 40 cases were found in 72 patients without optic canal fracture.Totally,283 sites of adjacent structure fractures were discovered with the most frequent sites of orbital apex region,accounting for 44.5%.Conclusion HRCT could well display optic canal fractures and accom- panied fractures of adjacent structures,providing important informations for diagnosis and intervention of traumatic visual loss.(Oph- thalmol CHN,2007,16:326-329)