Anesthesia management in a pediatric patient with complicatedly difficult airway: A case report.

BACKGROUND: Reports on perioperative anesthesia management in pediatric patients with difficult airways are scarce. In addition to relatively more difficulties in the technique of endotracheal intubation, the time for manipulation is restricted compared to adults. Securing the airways safely and avoiding the occurrence of hypoxemia in these patients are of significance. CASE SUMMARY: A 9-year-old boy with spastic cerebral palsy, severe malnutrition, thoracic scoliosis, thoracic and airway malformation, laryngomalacia, pneumonia, and epilepsy faced the risk of anesthesia during palliative surgery. After a thorough preoperative evaluation, a detailed scheme for anesthesia and a series of intubation tools were prepared by a team of anesthesiologists. Awake fiberoptic intubation is the widely accepted strategy for patients with anticipated difficult airways. Given the age and medical condition of the patient, we kept him sedated with spontaneous breathing during endotracheal intubation. The endotracheal intubation was completed on the second attempt after the failure of the first effort. Fortunately, the surgery was successful without postoperative complications. CONCLUSION: Dealing with difficult airways in the pediatric population, proper sedation allows time to intubate without interrupting spontaneous breathing. The appropriate endotracheal intubation method based on the patient's unique characteristics is the key factor in successful management of these rare cases.

[Effects of recombinant gene lentivirus containing LIM mineralization protein-1 on proliferation effect and expression of bone marrow mesenchymal stem cells in rats].

OBJECTIVE: To explore method of recombinant gene lentivirus containing LIM mineralization protein-1 (LMP-1) in transfecting bone marrow mesenchymal stem cells (BMSC), and to observe the effect of gene LMP-1 on proliferation effect and expression of BMSC. METHODS: Six clean SD rats aged 4 weeks were selected, bone marrow mesenchymal stem cells were extracted under sterile conditions and cultured to the third generation, then divided into three groups:control group (the third generation of BMSC), lentiviral vector transfection group (PGC-FU-GFP and Polybrene were injected into the third generation of BMSC) and recombinant gene transfection group (PGC-FU-LMP-1-GFP and Polybrene transfection were injected into the third generation of BMSC). After 48 hours' transfection, fluorescent expression were detected under immuno-fluorescence microscopy; lentiviral transfection efficiency were detected by flow cytometry; effect of lentiviral transfection on BMSC were evaluated by MTT; gene expression of transfected cells were determined by Western Blot. RESULTS: 1) The third generation of BMSC was cultured successfully,and transfected with MOI:100. After 48 hours, green fluorescent expression were detected and transfection efficiency was 67% under immuno-fluorescence microscopy; 2) Compared to control group, there were no statistical differences between control group and other two groups; 3) Western blot teast showed that 72KDa specific band was observed in recombinant gene transfection group and its size was similar to LMP-1 fusion protein (50 kDa+28 kDa=78 kDa). CONCLUSION: There is no effect of recombinant gene lentivirus containing LIM on BMSC, and can effectively influence the expression of LMP-1.

Inhibition of human prostate cancer xenograft growth by 125I labeled triple-helix forming oligonucleotide directed against androgen receptor.

BACKGROUND: The failure of hormone treatment for advanced prostate cancer might be related to aberrant activation of the androgen receptor. We have shown that (125)I labeled triple-helix forming oligonucleotide (TFO) against the androgen receptor gene inhibits androgen receptor expression and cell proliferation of LNCaP prostate cancer cells in vitro. This study aimed at exploring the effects of the (125)I-TFO on prostate tumor growth in vivo using a nude mouse xenograft model. METHODS: TFO was labeled with (125)I by the iodogen method. Thirty-two nude mice bearing LNCaP xenograft tumors were randomized into 4 groups and were intratumorally injected with (125)I-TFO, unlabeled TFO, Na(125)I and normal saline. Tumor size was measured weekly. The tumor growth inhibition rate (RI) was calculated by measurement of tumor weight. The expression of the androgen receptor gene was performed by RT-PCR and immunohistochemical study. The prostate specific antigen (PSA) serum levels were measured by enzyme linked immunosorbent assay. The tumor cell apoptosis index (AI) was detected by TUNEL assay. RESULTS: Tumor measurements showed that tumor development was significantly inhibited by either (125)I-TFO or TFO, with tumor RIs of 50.79% and 32.80% respectively. (125)I-TFO caused greater inhibition of androgen receptor expression and higher AIs in tumor tissue than TFO. Both the tumor weight and the PSA serum levels in (125)I-TFO treated mice ((0.93 +/- 0.15) g and (17.43 +/- 1.85) ng/ml, respectively) were significantly lower than those ((1.27 +/- 0.21) g and (28.25 +/- 3.41) ng/ml, respectively) in TFO treated mice (all P < 0.05). Na(125)I did not significantly affect tumor growth and androgen receptor expression in tumor tissue. CONCLUSIONS: The (125)I-TFO can effectively inhibit androgen receptor expression and tumor growth of human prostate cancer xenografts in vivo. The inhibitory efficacy of (125)I-TFO is more potent than that of TFO, providing a reference for future studies of antigen radiotherapy.

[Effects of anti-gene and antisense therapeutics on human prostate cancer xenograft in nude mice].

OBJECTIVE: To investigate the effects of triple-helix forming oligonucleotide (TFO) and antisense oligonucleotide (ASO) on androgen receptor (AR) expression and tumor growth of human prostate cancer xenografts in nude mice. METHODS: Thirty-two nude mice were inoculated with human prostate cancer cells of the line LNCaP-C4-2 were randomized into 4 equal groups: TFO treatment group, undergoing intra-tumor injection of TFO at the dose of 25 mgxkg(-1)x(2d)(-1) for 14 times, ASO treatment group, undergoing intra-tumor injection of ASO at the same dose for 14 times, SCO group, undergoing intra-rumor injection of sequence control oligonucleotide (SCO) at the same dose for 14 times, and control group. The body weight and xenograft tumor volume of the nude mice were monitored during the therapy. 28 days later venous blood samples were collected to measure the level of prostate specific antigen (PSA) by radioimmunoassay and then the mice were killed with their tumors taken out to measure the weight, and RT-PCR, immunohistochemistry, and radioligand binding assay were used to detect the AR gene mRNA and protein expression in the tumor tissues. RESULTS: By the end of experiment the volumes and weights of tumor of the ASO and ASO groups were all significantly lower than those of the control group (all P < 0.01) with the inhibition rates of 67.55% and 41.06% respectively, and the volumes and weights of tumor of the TFO group were all significantly lower than those of the ASO group (all P < 0.05). The tumor weight and AR expression levels of TFO group were significantly lower than those of ASO group (P < 0.05). The serum PSA level of TFO group was (6.6 +/- 1.0) ng/ml, significantly lower than that of the ASO group [(19.8 +/- 3.7) ng/ml, P < 0.05]. The mRNA and protein expression levels of AR of the TFO group were significantly lower than those of the other groups (all P < 0.05). There were no significant differences in all the above mentioned markers between the SCO and control groups (all P > 0.05). CONCLUSION: TFO shows significantly higher inhibitory effect on AR expression and tumor growth of human prostate cancer xenograft than ASO, and is a promising agent for prostate cancer gene therapy.

[Effect of antibody-targeted chemotherapy with pingyangmycin on prostate cancer cells in vitro].

OBJECTIVE: To investigate the effect of antibody-targeted chemotherapy against human prostate cancer LNCaP cells in vitro. METHODS: The monoclonal antibody 7E11C5.3 against human prostate cancer was conjugated to pingyangmycin (PYM), mediated by dextran T-40, and the immunoreactivity of 7E11C5.3 was determined by indirect enzyme-linked immunosorbent assay. The bacteriostatic activity of the conjugate was determined using TTC assay, and its cytotoxicity against LNCaP cells was determined by MTT assay. RESULTS: The 7E11C5.3:PYM molar ratio was l:54 in the conjugate, and the immunoreactivity of 7E11C5.3 was decreased by approximately 10% to 20% after conjugation. The bacteriostatic activity of conjugated PYM was 25% of that of free PYM. The 50% inhibitory doses (IC50) of 7E11C5.3-PYM conjugate and free PYM against the in vitro cultured LNCaP cells were 9.41-/+1.98 microg/ml and 29.92-/+7.88 microg/ml, respectively. CONCLUSION: 7E11C5.3-PYM conjugate displays stronger cytotoxicity against anti-prostate cancer effects than free PYM.

[Expression, renaturation and activity of BAC5-scFv expressed as inclusion body in E.coli].

AIM: To study the renaturation, purification and binding activity of scFv of anti-nasopharyngeal carcinoma monoclonal antibody(mAb) BAC(5) expressed as inclusion body in E.coli. METHODS: The E.coli BL21(DE3) transformed with the pET 22b-scFv was cultured and pulvereged by ultrasonic cell disintegrator. The collected inclusion bodies were denatured with 8 mol/L urea and renatured by dilution refolding, step dialysis and gel filtration chromatography. Binding activity of renatured BAC(5)-scFv was determined by immunohistochemical staining and Western blot. RESULTS: BAC(5)-scFv purified though Ni-NTA His Bind chromatographic clomn showed high purity. The highest proteins recovery rate was obtained through gel filtration chromatography. It was proved by Western blot and immunocytochemical staining that the renatured BAC(5)-scFv protein could specifically bind to CNE2 cells. CONCLUSION: BAC(5)-scFv expressed as inclusion body retained good activity after being dissolved, purified and renatured, which paves the way for preparing large amount of BAC(5)-scFv to be used for the study of radioimmunoimaging and therapy of nasopharyngeal carcinoma.

[Experimental study of combined target treatments using PYM-BAC5 and 131I-BAC5 to nasopharyngeal carcinoma].

BACKGROUND & OBJECTIVE: Combined therapy has been advocated for modern tumor treatment; the combined target therapy is a valuable research direction. Based on the previous research of nasopharyngeal carcinoma (NPC) radioimmunotherapy, this experiment was designed to develop two immunoconjugates of the monoclonal antibody BAC(5):PYM-BAC(5) and (131)I-BAC(5), and to assess the inhibition effects of their combined treatment on the NPC CNE-2 cells cultured in vitro. METHODS: Dextran T40 was used as media to link PYM and BAC(5). The conjugate PYM-BAC(5) was identified by testing its immunoactivity and the inhibition to mycobacterium. BAC(5) was labeled with (131)I by Chloramin-T method. Five experimental groups were set up:(1)PYM-BAC(5) group, (2)free PYM group, (3)(131)I-BAC(5) group, (4)(131)I-mIgG group, (5)the combined target treatment group ( (131)I-BAC(5)+PYM-BAC(5)). The antitumor effects of the five groups were assessed with MTT method. RESULTS: The 50% inhibition doses(IC(50)) of PYM-BAC(5) group and PYM group were 46.57 microg/ml and 316.7 microg/ml, respectively. The IC(50) of (131)I-BAC(5) group and (131)I-mIgG group to CNE2 were 4.42 x 10(5) Bq/ml and >11.1 x 10(5) Bq/ml,respectively. In the combined target treatment group(PYM-BAC(5)+(131)I-BAC(5)),the IC(50) of PYM-BAC(5) was 7.01 microg/ml and of (131)I-BAC(5) was 0.54 x 10(5) Bq/ml, which much less than other separate treatment groups. CONCLUSION: The inhibition effects of the target treatment ((131)I-BAC(5) and PYM-BAC(5)) on the NPC CNE-2 cells are stronger than non-target treatment (free PYM and (131)I-BAC(5)). The combined target treatment of the two immune ((131)I-BAC(5)+PYM-BAC(5)) conjugates gets stronger inhibition effects than their separate treatment.