OBJECTIVE:To observe the analgesic efficacy and adverse reactions of fentanyl transdermal in the treatment of advanced carcinoma pain.METHODS:43patients suffering from advanced carcinoma pain were administered with fentanyl transdermal system every3days with the dosage regulated accordingly until the patients feel painless or almost painless in24h.RESULTS:The general remission rate was90.7%for fentanyl transdermal in the treatment of moderate and severe cancer pain.Its main manifestations of adverse reactions were dizziness,drowsiness,nausea,vomiting and constipation,which have low incidence rates and degree of which were also low.CONCLUSION:Fentanyl transdermal system has durable and stable analgesic effect with its distinct way of transdermal slow-release,which is simple in operation and minor in adverse reactions;and it can be used as the substitutes for oral hadro-opioid.

Objective To study the mechanism of high intensity ultrasound (HIU) on COX-2 mRNA expression and apoptosis in breast cancer cell line MDA-MB-231 cells.Methods Human breast cancer cells MDA-MB-231 in vitro were exposed in HIU (50 w/cm2).RT-PCR was performed to detect the level of COX-2 mRNA before and after exposure.The ultrastructural changes in apoptotic cells were examined by transmission electron microscopy.Results The relative level of COX-2 mRNA decreased gradually along with the increase of exposure time and apoptotic bodies in MDA-MB-231 cells considerably increased under electron microscope.When exposure time was increased to 30 seconds,a few cells died.Conclusion HIU promotes apoptosis of breast cancer MDA-MB-231 cells in a COX-2 mRNA depended way.

The immune effect of two recombinant protein fragments of spike protein in severe acute respiratory syndrome coronavirus (SARS CoV) was investigated in Balb/c mice. Two partial spike gene fragments S1 (322 1464 bp) and S2 (2170 2814 bp) of SARS coronavirus were amplified by RT-PCR, and cloned into pET-23a prokaryotic expression vector, then transformed into competent Escherichia E. coli BL21 (DE3)(pLysS) respectively. Recombinant proteins were expressed and purified by Ni2+ immobilized metal ion affinity chromatography. The purified proteins mixed with complete Freund adjuvant were injected into Balb/c mice three times at a two-week interval. High titer antibody was detected in the serum of immunized Balb/c mice, and mice immunized with S1 protein produced high titer IgG1, IgG2a, IgG2b and IgG3, while those immunized with S2 protein produced high titer IgG1, IgG2a, but lower titer IgG2b and IgG3. Serum IFN-concentration was increased significantly but the concentrations of Il-2, IL-4 and IL-10 had no significant change. And a marked increase was observed in the number of spleen CD8+ T cells. The results showed that recombinant proteins of SARS coronavirus spike protein induced hormonal and cellular immune response in Balb/c mice.

BACKGROUND: Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) gene showed low expression in glioma cells. LRIG1 gene overexpression significantly enhanced LRIG1 mRNA and protein expression and inhibited its biological behavior. However, very few researchs are reported from the stand-point of inhibition of LRIG1 gene expression. OBJECTIVE: To construct specific RNA interference plasmids for LRIG1, establish stably transfected human glioma GL15 cell line, and observe its effect on expression of target gene LRIG1. METHODS: Designed and synthesized two shRNAs (named LRIG1-shRNA1 and LRIG1-shRNA2) specific for LRIG1 mRNA according to the GenBank, and one scrambled shRNA sequence as negative control, named pGenesil2-negative shRNA. The shRNA was inserted into pGenesil2 vector and sequenced. The recombinant vectors were transformed into E. coli. Picked up the positive clones and extracted the plasmids, which were transfected into GL15 cells by Metafectine. G418 was applied to select the stably transfected cell clones. Western Blotting was performed to examine the LRIG1 protein level.RESULTS AND CONCLUSION: The recombinant plasmids which contain shRNA were analyzed by restriction endonuclease digestion and DNA sequence, and it was proved that the fragment was inserted into the expected sites. Compared with the negative control group, the level of LRIG1 protein expression in pGenesil2-LRIG1-shRNA1(LRIG11) transfected cells and in pGenesil2-LRIG1-shRNA2(LRIG12) transfected cells was decreased by 47.9% (P < 0.01) and 32.8% (P > 0.05). The results confirmed that RNAi expressing vector specific for LRIG1 gene (pGenesil2-LRIG1-shRNA1) was successfully constructed, and the stable cell clones transfected with the shRNA expression vector showed inhibition of the expression of LRIG1 in glioma cell line GL15.

To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed into Escherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-beta-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0.992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.
Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Membrane Proteins/*biosynthesis ; Membrane Proteins/genetics ; Membrane Proteins/isolation & purification ; Plasmids/biosynthesis ; Plasmids/genetics ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics ; Recombinant Proteins/isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus/chemistry ; SARS Virus/*genetics ; Viral Vaccines/biosynthesis

Objective To investigate the effect of the pregnancy factor on the line drawn between the highest points of the two iliac crests ( T line) corresponding to the vertebral level in a multicenter clini?cal comparative study. Methods Hospitalized patients selected from the obstetric department or gynecolog?ical department, of American Society of Anesthesiologists physical statusⅠorⅡ, were divided into preg?nancy group ( group P ) and non?pregnancy group ( group NP ) . The patients were placed in the lateral posi?tion with their back vertical to the bed surface, the patient′s thighs were at an angle of approximately 90 de?grees to the trunk, and hip flexion was employed by flexing the patient′s knees to the chest. To determine the highest points of the two iliac crests, a line ( T line) was drawn between the highest points using a wire?reinforced epidural catheter. And another vertical line ( T′line) was made between the highest point of the iliac crest on the upper side ( not the side in the lateral position) and the ground. Ultrasonography was per?formed to identify and record the level of T line and T′line corresponding to the spinous process and lumbar interspace. Results A total of 1 763 cases completed the study, and there were 905 cases in group P, and 858 cases in group NP. Compared with group NP, the rate of T line at L3 spinous process and L3,4 in?terspace was significantly increased in group P ( P<0.05) . Compared with T′line, the rate of T line at L2,3 interspace and L3 spinous process was significantly decreased, and the rate of T line at L4 spinous process, L4,5 interspace and L5 spinous process was significantly increased in group P, and the rate of T line at L3 spinous process, L2,3 interspace and L3,4 interspace was significantly decreased, and the rate of T line at L4 spinous process and L4,5 interspace was significantly increased in group NP (P<0.01). Conclusion The level of T line corresponding to the vertebral level is significantly higher in the pregnant patients than in the nonpregnant patients.

Objective To investigate the expression of CXCR7 in several cutaneous malignant tumors including cutaneous squamous cell carcinoma (SCC),basal cell carcinoma (BCC) and invasive cutaneous malignant melanoma and their cell lines,as well as its significance.Methods Tissue specimens were obtained from the lesions of 30 patients with cutaneous squamous cell carcinoma,25 patients with basal cell carcinoma and 30 patients with cutaneous malignant melanoma.Immunohistochemistry was performed to detect the expression of CXCR7 protein in these tissue specimens and several cell lines (A375 human melanoma cells,M14 human melanoma cells,A431 human epidermoid carcinoma cells,HaCaT human keratinocytes).The mRNA expression of CXCR7 in these cell lines was measured by reverse transcription PCR.Results CXCR7 protein was apparently expressed in invasive cutaneous malignant melanoma.The high expression rate of CXCR7 protein was significantly elevated in cutaneous malignant melanoma tissue specimens compared with SCC and BCC tissue specimens [80％ (24/30) vs.26.67％ (8/30) and 8％ (2/25),x2 =17.16,28.36,both P ＜ 0.05].CXCR7 mRNA was expressed in A375,M14 and A431 cells,but not in HaCaT cells,with the strongest expression observed in A375 cells.Immunohistochemistry revealed the expression of CXCR7 protein only in A375 cells.Conclusions CXCR7 is highly expressed in cutaneous malignant melanoma and A375 cells,which may be involved in the malignant invasion and metastasis of melanoma.

OBJECTIVE: To compare the dosimetric differences of dosiology between intensity-modulated arc radiotherapy (IMAT) and dynamic intensity-modulated radiation therapy (dIMRT) in nasopharyngeal carcinoma. METHODS: CT data from 25 patients treated in our radiotherapy center were selected randomly for this study. For each patient, the IMAT technique and the fixed beam dIMRT technique were accomplished by the simultaneously integrated boost. Dose volume histogram (DVH) data, isodose distribution, monitor units (MUs) and treatment time were compared in the two techniques. RESULTS: There was no significant difference between the IMAT and the dIMRT in dose received by 95% of target volumes (D(95)) (P>0.05). Overall, the mean dose (D(mean)), maximal dose (D(max)) and volume percentage receiving at least of 107% of the prescribed dose (V(107%)) of planning target volume (PTV) for the IMAT were increased slightly ,compared with the dlMRT (P<0.05). There were no significant differences in dosimetric indices of organs at risk (OARs) including spinal cord,optical nerves,lens and temporomandibular joints in the two techniques (P>0.05). Compared with the dlMRI, the D(max) of brain stem for the IMAT was increased slightly (P<0.05). Similar trends was observed for the D(mean) and dose received by 50% of volume (D(50)) of the left and right parotid glands (P<0.05). Healthy tissue (defined as the volume of the body minus PTV,B-P) irradiated from 800 cGy in the IMAT was higher, and that from 1200-4500 cGy was lower compared with the dlMRI (P<0.05).The average number of MUs was reduced by 62.7% per fraction, and the treatment time was on average reduced by 60.1% per fraction in the IMAT compared with the dlMRI. CONCLUSION There is a slight difference in dosiology between the two radiotherapy techniques investigated, but they both meet the clinical requirement. Compared with the dIMRT, the IMAT delivers less irradiation to healthy tissue, uses fewer MUs and takes less time during radiotherapy for nasopharyngeal carcinoma.
Carcinoma, Squamous Cell ; radiotherapy ; Dose Fractionation, Radiation ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; radiotherapy ; Radiometry ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; methods ; Radiotherapy, Intensity-Modulated ; methods

The immune effect of two recombinant protein fragments of spike protein in severe acute respiratory syndrome coronavirus (SARS CoV) was investigated in Balb/c mice. Two partial spike gene fragments S1 (322-1464 bp) and S2 (2170-2814 bp) of SARS coronavirus were amplified by RT-PCR, and cloned into pET-23a prokaryotic expression vector, then transformed into competent Escherichia E.coli BL21 (DE3)(pLysS) respectively. Recombinant proteins were expressed and purified by Ni2+ immobilized metal ion affinity chromatography. The purified proteins mixed with complete Freund adjuvant were injected into Balb/c mice three times at a two-week interval. High titer antibody was detected in the serum of immunized Balb/c mice, and mice immunized with S1 protein produced high titer IgG1, IgG2a, IgG2b and IgG3, while those immunized with S2 protein produced high titer IgG1, IgG2a, but lower titer IgG2b and IgG3. Serum IFN-γ concentration was increased significantly but the concentrations of IL-2, IL-4 and IL-10 had no significant change. And a marked increase was observed in the number of spleen CD8+ T cells. The results showed that recombinant proteins of SARS coronavirns spike protein induced hormonal and cellular immune response in Balb/c mice.

Objective To construct a prokaryotic expression system containing the dense granule protein 4(GRA4) of Toxoplasma gondii,purify the expressed protein and detect its immunogenicity.Methods The specific fragment of GRA4 gene was amplified by PCR.After subcloning the prokaryotic expression recombinant pET,GRA4,the expressed product was purified with His?BindTM affinity chromatography and analyzed by Western blot.BALB/c mice were immunized with the GRA4 recombinant protein,and the antibody IgG titer was detected by ELISA.Results The pET,GRA4 prokaryotic expression system was obtained.The MW of the expressed protein was Mr 40 000 and formed in inclusion body.After purification,the recombinant protein could be specifically recognized by the T.gondii infected rabbit serum.Mice immunized with the purified recombinant protein elicited high titer of IgG antibody.Conclusion The pET,GRA4 recombinant protein was successfully expressed and purified,which shows the immunogenicity.