Effect evaluation of repeated debridement after endoscopic sinus surgery.

OBJECTIVE: To investigate the effect of early phase debridement by the different intervention frequencies on postoperative symptoms recovery and turnover of mucosa after functional endoscopic sinus surgery (FESS). METHODS: 67 patients undergone FESS were divided into intervention group and control group. Intranasal corticosteroids, macrolides antibiotics and postoperative saline douching were used in both groups. Debridement was performed on the 1(st), 4(th), 8(th) postoperative week on patients of invention group, while once per week on patients of control group. The primary outcome measure was visual analogue scale (VAS) and Lund-Kennedy Endoscopic Score (LKES) Results: On the 4(th) week, the control group presented more release on nasal block, the VAS of the two groups is 3.45 ± 1.16 and 4.83 ± 1.47 in the control group and intervention group respectively which was significantly different. The LKES on crust decreased more in the control group (1.12 ± 0.64 in the control group and 1.90 ± 0.47 in the intervention group). However, the control group complained more sever facial pain and uncomfortable; the VAS of two groups is 5.92 ± 0.91 and 2.74 ± 1.41 respectively. On the 8(th) week, there were no significant difference between the two groups on all domains of VAS and LKES except lower scar was shown in the control group. CONCLUSIONS: Benefit of frequent debridement during the early postoperative was not in positive correlation with patients recovering from ESS. Excessive debridement may induce more surgical trauma and cause more facial pain to patients. Therefore, in terms of subjective recovery and health care costs, appropriate extending postoperative management time and decreasing intervention frequencies will not affect the therapeutic effect of endoscopic surgery for chronic sinusitis.

[Culture, identification and label of embryonic rat neural stem cells].

OBJECTIVE: To explore the methods of culture, identification and label of embryonic rat neural stem cells. METHOD: The cells isolated from fetal rat hippocampus were identified with nestin immunocytochemical fluorescent staining. The cellular multiplication was observed by immunocytochemical fluorescence co-label after accession of BrDU. The neural stem cells (NSCs) were marked with fluorescent dye, bisbenzimide (Hoechest33342) and induced to differentiate. The differentiated cells were detected with Neuron Specific Enolase (NSE) and Glial Fibrillary Acidic Protein (GFAP) immunocytochemical fluorescent staining respectively. RESULT: Nest-like clusters of neural stem cells were obtained in suspension and the cells could be differentiated into neurons and astrocytes which maintaining the main characteristics of NSCs after 8 passages of culture. The label efficiency of cells with Hoechest33342 was 97% and no attenuation of fluorescent brightness was observed after 8 passages of culture. The cellular fluorescence was observed in the NSCs and the differentiated cells. CONCLUSION: The cells from embryonic rat hippocampus possessed the abilities of division, multiplication and self-renew, which were believed to be the main characteristics of NSCs of the central nervous system. The cells could be efficiently labeled with fluorescent dye and could be used as donor cells in experimental research on NSCs transplantation.

[Neurofibromatosis type 2].

OBJECTIVE: To recognise the predisposing factors, clinical manifestations, diagnosis and treatment of neurofibromatosis type 2 (NF2). METHOD: The clinical data of one NF2 case was reported and the literatures were also reviewed. RESULT: The patient was diagnosed at a much later stage than onset. Progressive hearing loss and tinnitus were the initial symptoms. MRI scan indicated space-occupying lesions in the bilateral cerebellopontine angles, bilateral cavernous sinuses and bilateral cervical parts of the patient. The patient was diagnosed as NF2 according to the National Institutes of Health (NIH) criteria, and received operation on the left acoustic tumor. The tumor was proved to be schwannomas by pathological test. The hearing loss and the facial nerve paralysis (House-Brackmann II) had appeared after operation. CONCLUSION: NF2 is an autosomal dominant, highly penetrant disease which is characterized by bilateral vestibular schwannomas. Early diagnosis and management for tumor is very important for survival and hearing preservation. The "golden standard" in terms of diagnostic precision is the magnetic resonance imaging (MRI) scan with gadolinium enhancement.

[Expression of perlecan in laryngeal carcinoma cell and its significance].

OBJECTIVE: To study the expression of perlecan in human laryngeal carcinoma cells and its significance. METHOD: The expression of perlecan mRNA in human laryngeal carcinoma cells, Hep-2 cells were investigated by using the techniques of semi-quantify RT-PCR. The expression of perlecan in the Hep-2 cells was investigated by using the techniques of immunohistochemistry. RESULT: It showed that the expression level of perlecan and perlecan mRNA significantly increased in Hep-2 cells as compared with the normal cells, Hacat cells (P < 0.01). CONCLUSION: These data raise the possibility that perlecan many play key roles in the growth,invasion and metastasis of human laryngeal carcinoma cells through either a paracrine or an autocrine mechanism.

[Suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA].

OBJECTIVE: To study the suppression of proliferation of Hep-2 cells by stable expression of anti-sense perlecan cDNA. METHOD: In this study, the plasmids of recombination eukaryotic expression vector perlecan anti-sense cDNA (pAP) were transfected into Hep-2 cells by using cationic liposome (lipofectamine 2000) and divided into three groups: non-transfected group, WT group; transfection with no load carrier, neo group; and transfection with the pAP plasmid, pAP group. Semi quantify RT-PCR, western blot assay and MTT assay were used to detected the expression of perlecan mRNA and protein in the three groups; the level of cell proliferation; and the responsivity of basic fibroblast growth factor (bFGF). RESULT: It was showed that the expression of perlecan mRNA and protein were significantly reduced in the pAP group compared with WT group and ph beta Apr-neol transfected group ( P < 0.01). In the presence of 1 microg/L of bFGF in low serum (0.1% FCS), the pAP transfected cells showed a reduced proliferation rate (MTT assay) while the wild type cells and ph beta Apr-neol transfected cells grew rapidly. CONCLUSION: The growth of Hep-2 cells could be inhibited significantly by perlecan anti-sense cDNA plasmids transfection.

[Expression of vascular endothelial growth factor and its receptors in laryngeal carcinoma cell and its significance].

OBJECTIVE: To study the expression of vascular endothelial growth factor (VEGF) and both the Flt-1 and KDR high affinity VEGF receptors in human laryngeal carcinoma cells and its significance. METHOD: In this study, we investigated the expression of VEGF mRNA and both the Fit-1 mRNA and KDR mRNA high affinity VEGF receptors in human laryngeal carcinoma cells using techniques of semi-quantify RT-PCR. RESULT: It was showed that the expression level of VEGF mRNA was significantly increased in human laryngeal carcinoma cells as compared with the normal cells, Hacat cells (P <0.05), and there was the high expression level of Flt-1 mRNA in the Hep-2 cells, but not KDR mRNA. CONCLUSION: These data raise the possibility that VEGF and its receptors many play key roles in the growth, invasion and metastasis of human laryngeal carcinoma cells through either a paracrine or an autocrine mechanism.

[Construction of recombinant adenovirus containing human Bcl-2 and its expression in the spiral ganglion cells].

OBJECTIVE: To construct the adenoviral vector containing human Bcl-2 gene and to study the expression of the gene in the spiral ganglion cells (SGC) in vitro. METHODS: Human Bcl-2 cDNA obtained from the plasmid pUC-CAGGS/Bcl-2 was cloned into the plasmid pAdTrack-CMV. Then, pAdTrack/Bcl-2 was cotransferred with adenoviral backbone vector into E. coli strain BJ5183. The recombinant adenoviral plasmid was identified by restriction analysis with Pac I and transfected into HEK293 cells to package and amplify recombinant adenovirus particles which would be identified by Electron microscope. After the adenovirus infected the rat spiral ganglion cells, the expression of Bcl-2 gene was detected by Western Blot and RT-PCR. RESULTS: The recombinant AdEGFP/Bcl-2 plasmid was correctly constructed and confirmed by restriction endonuclease analysis. The viral particles in HEK293 cells were identified by Electron microscope. RT-PCR and Western blot showed that Bcl-2 gene was exactly transcripted and expressed in transgene SGC. CONCLUSIONS: The method based on homologous recombination in bacteria is simple and high efficient. The recombinant adenoviral vector containing human Bcl-2 cDNA was constructed and the transgene SGC expressed human Bcl-2 gene in vitro successfully. It provided foundation for the further study of protection for the impaired SGC by hBcl-2 gene.