Application of the Mathieu combined tunnel technique for repairing glans dehiscence after failed hypospadias repair / 亚洲男科学杂志(英文版)

Repairing glans dehiscence after failed hypospadias repair is challenging for pediatric surgeons. Here, we introduced and evaluated a newly modified Mathieu technique, Mathieu combined tunnel (MCT), which involves multiple custom-designed flaps for the shortage of flap source material after repeated operations; we also constructed a tunnel to avoid the glans incision that may carry new risks of dehiscence. This retrospective study included 26 patients who were consecutively admitted to the First Affiliated Hospital of Sun Yat-Sen University (Guangzhou, China) for glans dehiscence repair after failed hypospadias repair from October 2014 to October 2020; sixteen patients underwent surgery using the MCT (MCT group) and ten patients underwent surgery using the tubularized incised plate (TIP) technique (TIP group). The operative time, blood loss, postoperative complications, normal urethral meatus rate, success rate, and Hypospadias Objective Penile Evaluation (HOPE) score were compared between the two groups. The MCT group achieved an overall satisfactory penile appearance and voiding function, with a higher rate of normal urethral meatus (15/16, 93.8%) and a lower rate of glans dehiscence (1/16, 6.2%), compared with the TIP group (70.0% and 30.0%, respectively). However, these differences were not statistically significant, possibly because of the limited number of patients (all P > 0.05). Mean postoperative HOPE scores were similar in the MCT group (mean ± standard deviation: 8.83 ± 0. 89) and TIP group (8.94 ± 0.57) (P > 0.05). No significant differences were found between the two groups in terms of blood loss and success rate, nor in the rates of various complications (e.g., fistula, urethral stricture, and glans dehiscence). In conclusion, the MCT technique appears to be feasible and reliable for repairing glans dehiscence after failed hypospadias repair.

Diagnosis and treatment of ejaculatory duct obstruction: Current status and advances / 中华男科学杂志

Ejaculatory duct obstruction (EDO) is one of the obstructive factors for 1－5% of all cases of male infertility and it is, however, surgically correctable. Congenital developmental abnormality is a most common cause of EDO. The clinical manifestations of EDO are varied, typically with the decline of four semen parameters. Transrectal ultrasonography is an important imaging method for the diagnosis of EDO and guidance in its surgery. MRI provides high-resolution images of the reproductive system as evidence. Transurethral resection of the ejaculatory duct (TURED) is a classical operation, the application of transurethral seminal vesiculoscopy has become a new trend of minimally invasive surgery in the treatment of EDO, and the latest flexible vesiculovasoscopy (FVV) or vasoscopy techniques may further improve the diagnosis and treatment of EDO.

Silencing of endothelin-1 suppresses growth, migration, and invasion of nasopharyngeal carcinoma cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the role of endothelin-1 (ET-1) gene in regulating the proliferation, migration and invasion of nasopharyngeal carcinoma cells.</p><p><b>METHODS</b>A lentivirus-mediated shRNA-ET-1 vector was infected into 5-8F cells, and the interference efficiency was examined with Western blotting. MTT assay, cell cycle analysis, plate colony formation assay, Transwell assay, Boyden chamber assay and tumor growth assay were carried out to analyze the changes in cell proliferation, migration and invasion. The expressions of genes related with epithelial-mesenchymal transition (EMT) were examined using Western blotting.</p><p><b>RESULTS</b>shRNA-ET-1 transfection significantly inhibited the expression of ET-1, and suppressed the growth, migration and invasion of 5-8F cells. ET-1 knockdown enhanced the expression of E-cadherin and CK18 and inhibited the expression of N-cadherin and vimentin.</p><p><b>CONCLUSION</b>ET-1 promotes cell growth, migration and invasion by modulating the genes associated with epithelial-mesenchymal transition in nasopharyngeal carcinoma cells.</p>

The lipid-regulating effect and safety of combined statin and bezafibrate therapy in acute coronary syndrome patients complicating with dyslipidemia / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the lipid-regulating effect and safety of combined statin and bezafibrate therapy in acute coronary syndrome(ACS) patients complicating with dyslipedemia.</p><p><b>METHODS</b>One hundred and four hospitalized patients with established ACS and increased serum triglycerides (TG) levels and/or low serum levels of high density lipoprotein cholesterol (HDL-C) were selected. Except for conventional therapy, the patients were randomly divided into 2 groups: control group (n = 52), treated with atorvastatin 20 mg qn or other statin equivalent to 20 mg atorvastatin; treatment group (n = 52), treated with the same dose statin plus bezafibrate 200 mg bid. The serum levels of total cholesterol (TC), TG, low-density lipoprotein cholesterol (LDL-C) and HDL-C were assessed before and after 6 and 12 weeks treatment, side effects and adverse events were recorded.</p><p><b>RESULTS</b>After 6 weeks treatment, the serum levels of TC, TG and LDL-C in two groups were significantly reduced compared to baseline (all P < 0.05), which were further declined after 12 weeks treatment, and the reduction was more significant in treatment group(29.8%, 38.0% and 36.1%, respectively) than in control group(14.7%, 9.8% and 26.7%, respectively) (all P < 0.05). After treatment, the serum levels of HDL-C in the two groups were significantly higher than the baseline levels, especially after 12 weeks treatment (all P < 0.05), and the elevations of HDL-C levels in control group and in treatment group were 19.3% and 24.2%, respectively, but there were no significant difference between the two groups (P > 0.05). After 12 weeks, the rates reaching to target goals of LDL-C, TG, HDL-C, and non-HDL-C levels in the treatment group (69.2%, 88.5%, 92.3%, 46.2% and 65.4%, respectively) were significantly higher than those in the control group (34.6%, 65.4%, 46.2%, 7.7% and 42.3%, respectively, all P < 0.05). No serious side effects were observed in the two groups during the treatment period.</p><p><b>CONCLUSION</b>The combined statin and bezafibrate treatment is safe and could increase the ratios of reaching target lipid levels in ACS patients complicating with increased TG and (or) decreased HDL-C.</p>

Esophageal dynamic and laryngopharyngeal reflux play a role in pathogenesis of vocal cord polyps / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>Through monitoring esophageal dynamic change, and detection of laryngopharyngeal reflux(LPR) and gastroesophageal reflux events,to discuss the relationship of vocal cord polyps with laryngopharyngeal reflux.</p><p><b>METHODS</b>Thirty-two patients with vocal cord polyps were diagnosed by electronic laryngoscopy in Nanfang Hospital between October 2011 to May 2012. This study applied high-resolution esophageal manometry (HRM) and ambulatory 24-hour multichannel intraluminal impedance-pH monitoring (MII-pH) to obtain the upper esophageal sphincter(UES) and lower esophageal sphincter pressure, characteristics of sectional esophageal motility; laryngopharyngeal reflux (LPR)and gastroesophageal reflux events, as well as the reflux properties of substances. Sixteen healthy volunteers were recruited as normal controls.</p><p><b>RESULTS</b>UES relaxation duration, duration of UES relaxation time, UES relaxation recovery time and mean length of LES were all shorter than those of the control group (t were 2.244, 2.624, 2.310 and -2.397, P < 0.05). There were 40.6% (13/32) LPR and 50.0% (16/32) gastroesophageal reflux found in vocal polyp patients. Median number (M [P25; P75]) of laryngopharyngeal acid reflux events were 0.5[0.0;3.5] and 0.0[0.0;0.0] in vocal polyp group and the controls, median mean time of laryngopharyngeal acid exposure 0.1[0.0;1.7] and 0.0[0.0;0.0] min, median clearance time of laryngopharyngeal acid were 3.5[0.0;53.5] and 0.0[0.0;0.0] s, median scores of DeMeester were 14.8[1.6;31.3] and 1.8[1.1;4.1] and median frequency of total liquid reflux episodes were 46.5[25.3;69.0] and 32.5[20.0;36.3], respectively. The median numbers of laryngopharyngeal acid reflux events, time of acid exposure, time of acid clearance, DeMeester scores and frequency of total liquid reflux episodes were increased or higher in vocal polyp group than those in the controls (z were 2.481, 2.767, 2.767, 2.344 and 1.980, P < 0.05).</p><p><b>CONCLUSIONS</b>There are upper esophageal sphincter and Lower esophageal sphincter dismotility in vocal polyp patients with LPR. LPR events were dominated by acid reflux in upright position.Esophageal dynamic disfunction and LPR should be considered in the study of the pathogenesis of vocal cords polyps.</p>

Pathologic changes of the palatopharyngeal muscles in adult patients with obstructive sleep apnea hypopnea syndrome / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the pathology of palatopharyngeal muscle obtained from patients with obstructive sleep apnea hypopnea syndrome (OSAHS).</p><p><b>METHODS</b>The samples from both groups were studied under HE, nicotinamide adenine dinucleotide-tetrazolium reductase (NADH- TR), modified Gomori trichrome (MGT) and adenosine triphosphatase (ATPase) staining. There were 36 cases of OSAHS who received uvulopalatopharyngoplasty in the experimental group (including 6 mild, 6 moderate and 24 severe cases). There were 6 patients with chronic tonsillitis but without OSAHS as matched control group. Both groups were diagnosed by PSG.</p><p><b>RESULTS</b>Centralized located nuclei and obvious variability of the size of fiber types were observed in both groups. The occurrence rate of the former were 1/6 in control group and 52.8% (19/36) in OSAHS, while the rate of the latter were 4/6 and 58.3% (21/36)respectively. A large number of fibers in both groups (control group 5/6, OSAHS group 28/36) presented an irregularly distributed staining for oxidative activity reaction in NADH stain.Endomysium connective tissue proliferation, a lobular or motheaten appearance, target-like fibers, ragged red fiber (RRF) and muscle necrosis were only observed in OSAHS group.While it was more common in serious OSAHS patients. Dominance of type 1 fibers were observed in matched control group in ATPase stain. Clusters of type 2 fibers or clusters of both type fibers were observed in OSAHS, especially more common in serious OSAHS. There was a predominance of the type 2 fibers in some OSAHS patients.</p><p><b>CONCLUSIONS</b>The observation of HE and special muscular stain identified that palatopharyngeal muscle of OSAHS patients had pathological lesion. The pathological changes included muscular lesion and abnormal distribution of different fiber types, the rate of type 1 fiber which maintained the opening of upper air way decreased.</p>

Value of tonsillectomy in patient with obstructive sleep apnea hypopnea syndrome with Friedman staging I / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the availability of tonsillectomy in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) staged as Friedman I.</p><p><b>METHODS</b>Fifty-six patients with OSAHS in Friedman stage I who refused uvulopalatopharyngoplasty (UPPP) received tonsillectomy merely from January 2004 to March 2010. There were 20 mild, 24 moderate and 12 serious patients respectively in this group. The other 68 OSAHS patients in Friedman stage I received UPPP at the same time as matched group, including 26 mild, 28 moderate and 14 serious patients.</p><p><b>RESULTS</b>There was no significant difference before operation in terms of age, body mass index, apnea hypopnea index (AHI), the lowest pulse oxygen saturation (SPO(2)) and average SPO(2) between the two groups. There were significant difference in mean length of operation (U = 0.000, P < 0.01), hospitalization day (U = 458.5, P < 0.01), visual analogue scale after surgery (U = 0.000, P < 0.01) in these two group. There was no significant difference in surgical effective rate between the two groups (χ(2) = 0.857, P > 0.05). There was also no significant difference in terms of age, body mass index, AHI, the lowest SPO(2) and average SPO(2) after operation between the two groups (t test P > 0.05). The surgical effective rate for the long term of the two groups was equal (χ(2) = 0.857, P > 0.05). Even patients with serious OSAHS in Friedman stage I, the surgical effective rate of the two groups was equivalent (Fisher's exact test, P > 0.05).</p><p><b>CONCLUSIONS</b>Tonsillectomy is a safe and effective surgery for OSAHS in Friedman stage I, whose main structural load lies in the hypertrophic tonsil. It should be the first surgical choice for OSAHS in Friedman stage I.</p>

Effects of enhancer of zeste homolog (EZH2) downregulation on the proliferation and invasion of nasopharyngeal carcinoma cell and the possible mechanisms / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of the enhancer of zeste homolog 2 (EZH2) gene on cell growth and invasion of the nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>Recombinant lentivirus vector for shRNA delivery of EZH2 was constructed and transfected into 293FT cells. After collecting the viral particles, the NPC cell line 5-8F cells were transfected. The effects of EZH2 silence on cell proliferation and cell cycle were detected using MTT assay, plate colony formation assay and flow cytometry. The migration and invasion of 5-8F cells were determined by wound healing assay and matrigel invasion assay, respectively. The expressions of EZH2 and epithelial-mesenchymal transition (EMT)-related markers at mRNA and protein levels were examined by real-time PCR and Western blot respectively.</p><p><b>RESULTS</b>The expressions of EZH2 mRNA and protein in the transfected 5-8F cells were obviously reduced. MTT assay showed that EZH2 downregulation significantly inhibited the growth of 5-8F/shEZH2 cells (P < 0.001). Colony formation rate (84.44%) of 5-8F/shEZH2 cells was lower than control (31.56%, P = 0.001). Cell cycle analysis showed that most 5-8F/shEZH2 cells were arrested in G0/G1 phase, with a very low ratio of cells in S phase. Wound healing assay indicated that the migration ability of cells silencing EZH2 decreased significantly, and the 48-hour relative migration distance of 5-8F/ShEZH2 cells and control cells was 0.58 ± 0.05, and 0.81 ± 0.02, respectively (P < 0.000). Matrigel invasion assay, showed the invasive capacity of cells silencing EZH2 was significantly inhibited, with less penetrating cells (72.23 ± 4.08) compared to control (150.95 ± 16.27), P < 0.000. The mRNA expressions of epithelial markers E-cadherin and Keratin 18 in the cells silencing EZH2 increased by 177% and 158% respectively, and the mRNA expressions of mesenchymal markers β-catenin and N-cadherin decreased by 18.04% and 41.18% respectively. Similar results also were obtained with Western blot analysis.</p><p><b>CONCLUSION</b>EZH2 significantly enhanced the proliferation and invasion of nasopharyngeal carcinoma cells in vitro, which might be mediated by inducing EMT.</p>

Suppressing tumor growth of nasopharyngeal carcinoma by hTERTC27 polypeptide delivered through adeno-associated virus plus adenovirus vector cocktail / 癌症

Nasopharyngeal carcinoma(NPC) is a metastatic carcinoma that is highly prevalent in Southeast Asia. Our laboratory has previously demonstrated that the C-terminal 27-kDa polypeptide of human telomerase reverse transcriptase (hTERTC27) inhibits the growth and tumorigenicity of human glioblastoma and melanoma cells. In this study, we investigated the antitumor effect of hTERTC27 in human C666-1 NPC cells xenografted in a nude mouse model. A cocktail of vectors comprising recombinant adeno-associated virus (rAAV) and recombinant adenovirus (rAdv) that each carry hTERTC27 (rAAV-hTERTC27 and rAdv-hTERTC27; the cocktail was abbreviated to rAAV/rAdv-hTERTC27) was more effective than either rAAV-hTERTC27 or rAdv-hTERTC27 alone in inhibiting the growth of C666-1 NPC xenografts. Furthermore, we established three tumors on each mouse and injected rAAV/rAdv-hTERTC27 into one tumor per mouse. Although hTERTC27 expression could only be detected in the injected tumors, reduced tumor growth was observed in the injected tumor as well as the uninjected tumors, demonstrating that the vector cocktail could provoke an antitumor effect on distant, metastasized tumors. Further studies showed the observed antitumor effects included inducing necrosis and apoptosis and reducing microvessel density. Together, our data suggest that the rAAV/rAdv-hTERTC27 cocktail can potently inhibit NPC tumor growth in both local and metastasized tumors and should be further developed as a novel gene therapy strategy for NPC.

Enhancement of target gene expression by recombinant adeno-associated virus combined with recombinant adenovirus in vivo / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the changes in the intensity and temporal pattern of target gene expression in the tumor tissue of nude mice bearing human nasopharyngeal carcinoma (NPC) following injection of recombinant adeno-associated virus (rAAV) and recombinant adenovirus (AdV) in vivo.</p><p><b>METHODS</b>EBV-positive human NPC cell line C666-1 was inoculated subcutaneouly in nude mice. After the tumor mass reached 3 mm in diameter, 1.5 × 10(11) v.g (virus genome) rAAV-EGFP, 2.5 × 10(8) pfu rAdV-EGFP or their balanced mixture was injected intratumorally. At 5 and 10 days after the injection, the tumor tissues were harvested for immunohistochemical staining of GFP, and the ratio of the GFP-positive cells and the intensity of GFP expression was determined.</p><p><b>RESULTS</b>Immunohistochemistry for GFP showed that 5 days after the injection, GFP expression was detected (1.70 ∓ 0.48) in the tumor tissue in rAAV group, and the peak expression levels was seen in rAdV group (6.00∓1.94); the expression level was comparable between the combination group (6.90 ∓ 1.92) and rAdV group. At 10 days, GFP expression was considerably lowered to 2.00 ∓ 0.67 in rAdV group but increased to 8.00∓1.15 in rAAV group. The expression in the combination group maintained a high level at 10 days (10.10∓1.63), which was significantly higher than that in rAAV group (P%0.001).</p><p><b>CONCLUSION</b>Transfection with rAAV combined with rAdV allows instant, sustained and significantly enhanced expression of the target gene in the tumor tissue. This approach takes advantages of the two viruses and can be ideal for exogenous gene delivery into the tumor tissues.</p>

Effect of Epstein-Barr virus nuclear antigen 1 on cell proliferation and cell cycle in nasopharyngeal carcinoma cells / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the effect of Epstein-Barr virus nuclear antigen 1 (EBNA1) on cell proliferation and cell cycle in nasopharyngeal carcinoma (NPC) cells.</p><p><b>METHODS</b>Recombinant lentivirus that encoded EBNA1 short hairpin RNA (shRNA) was prepared. The C666-EBNA1 (CE) cells were transduced with lentivirus and selected by fluorescence activated cell sorting (FACS) to repress EBNA1 expression. The protein expression levels of EBNA1 were examined by Western blot. The effect of EBNA1 silence on cell proliferation was analyzed by MTT assay and cell growth assay, respectively. Cell cycle was assessed by flow cytometry. The mRNA and protein levels of cell cycle regulators were examined by real-time PCR and Western blot.</p><p><b>RESULTS</b>Recombinant lentivirus encoded EBNA1 shRNA was successfully constructed. The EBNA1 expression in CE cells was significantly reduced by lentivirus-mediated RNA interference. The results of cell counting and MTT assay showed that EBNA1 down-regulation significantly inhibited cell growth in CE-shRNA EBNA1 cells (P < 0.05). Compared with the control group, the percentage of cells in G0-G1 phase was increased from (62.43 ± 6.62)% to (89.66 ± 0.64)% (t = -7.091, P = 0.002), and that in S phase was decreased from (34.93 ± 7.36)% to (7.82 ± 2.44)% (t = 6.095, P = 0.004). The mRNA expressions of c-myc, CDK4, CDK6 and pRb were decreased by 65.60%, 34.06%, 41.05% and 55.29% respectively with the similar results in protein expression levels.</p><p><b>CONCLUSIONS</b>Suppression of EBNA1 may inhibit the growth of nasopharyngeal carcinoma cells in vitro and induce a G1-phase cell cycle arrest, which might be mediated by down-regulation of c-myc, CDK4, CDK6 and pRb.</p>

Application of SELDI-TOF-MS in establishing a model for predicting radiotherapy response of hypopharyngeal cancers / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the serum proteomic fingerprints in patients with hypopharyngeal squamous cell carcinoma (HPSCC) by surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) protein chip array technique.</p><p><b>METHODS</b>The serum samples were obtained from 58 HPSCC patients for protein expression analysis using SELDI-TOF Protein Chip technique and cation-exchange (CM10) protein array. All the spectra were compared and the qualified mass peaks with mass-to-charge ratios (m/z) between 1 and 70 kD were autotimatically detected. The tree analysis pattern was generated using Biomarker Patterns Software.</p><p><b>RESULTS</b>The protein profiles of HPSCC serum were analyzed according to the clinical and pathological features of the patients and their treatment response. No significant difference was noted in the serum proteins between HPSCC patients with different statuses of cervical lympha node metastasis (P>0.05), and the difference between well differentiated and poorly differentiated HPSCC was only minor. No significant difference was found in the serum proteins between chemotherapy-sensitive patients and the insensitive patients (P>0.05), but 5 proteins were identified to be overexpressed in the sensitive patients (P < / = 0.05). Radiotherapy-sensitive HPSCC patients were segregated from the insensitive group with a sensitivity of 86.67% and specificity of 100%.</p><p><b>CONCLUSION</b>The serum protein at the m/z value of 6115.74 is overexpressed in radiotherapy-sensitive HPSCC patients. Serum protein profiling allows the prediction of radiotherapy response in HPSCC patients, and the identified proteins may serve as candidate biomarkers for predicting the radiotherapy sensitivity of HPSCC.</p>

Silencing of COX-2 in nasopharyngeal carcinoma cells with a shRNAmir lentivirus vector / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a miR-155-based lentivirus vector to induce cyclooxygenase-2 gene silencing in nasopharyngeal carcinoma (NPC) cells by expressing anti-COX-2 shRNAmir.</p><p><b>METHODS</b>miR-155-based anti-COX-2 shRNAmir template was synthesized and inserted into pLVTHM plasmid. The recombinant pLVTHM/shRNAmir was transfected into 293FT cells for packaging the lentivirus vector. After infection with the lentivirus vector, the GFP-positive cells were screened by flow cytometry, and COX-2 mRNA level was detected by RT-PCR.</p><p><b>RESULTS</b>Restriction digestion and DNA sequencing confirmed successful construction of the anti-COX-2 vector pLVTHM/shRNAmir. A subline of C666-1 cells was established after infection with the lentivirus vector, and the COX-2 expression in the cells was stably silenced.</p><p><b>CONCLUSION</b>The shRNAmir lentivirus vector constructed may serve as an effective COX-2 inhibitor, which may facilitate future studies of gene therapy of NPC.</p>

Analysis of clinical characteristics of chronic rhinosinusitis in patients with postirradiated nasopharyngeal carcinoma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the characters of chronic rhinosinusitis in patients with irradiated nasopharyngeal carcinoma.</p><p><b>METHODS</b>There were 65 cases of chronic rhinosinusitis after irradiated nasopharyngeal carcinoma (NPC, experimental group) and 65 cases of common chronic rhinosinusitis (CRS, control group) in the study. The visual analogue scale (VAS) was used to evaluate the intensity of subjective symptoms. Endoscopic finding was recorded and CT results were evaluated by Lund-Mackay scoring system.</p><p><b>RESULTS</b>As to the VAS, nasal secretion was significantly more severe in experimental group (7.86+/-1.62), compared with control group (5.12+/-1.32, t=10.541, P<0.01). As to endoscopic finding, middle nasal meatus were clean in 35 (53.8%) cases in experimental group, and 23 cases (35.4%) in control group (chi2=4.483, P<0.05). CT score was (7.03+/-4.63) in experiment group, and (11.42+/-3.32) in control group (t=-6.207, P<0.05). The main reason lays in lower CT score and lower involved rate of ostiomeatal complex, frontal sinus, maxillary sinus, anterior ethmoid sinus.</p><p><b>CONCLUSIONS</b>The characters of chronic rhinosinusitis in patients with irradiated nasopharyngeal carcinoma is quite different from the common CRS and different therapeutic measures should be taken.</p>

Association of CXCR4 and SDF-1alpha with organ-specific metastasis of nasopharyngeal carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the role of CXCR4/SDF-1alpha axis in organ-specific metastasis of nasopharyngeal carcinoma (NPC) by assessment of CXCR4 expression in NPC cells and SDF-1alpha expression in distant target organs of NPC.</p><p><b>METHODS</b>Thirty patients with NPC and fifteen normal subjects were recruited in this study. The expressions of CXCR4 in NPC and normal cases were identified by RT-PCR and immunohistochemistry (IHC), then the relationship between CXCR4 expression and clinicopathological factors was analyzed. IHC was also used to analyze the SDF-1alpha protein expression in normal cervical lymph nodes (including normal superior and inferior deep cervical lymph nodes), bone marrow, lung, liver, kidney and colon tissues of NPC patients (5 cases/each group).</p><p><b>RESULTS</b>The relative expression level of CXCR4 mRNA in NPC (0.71 +/- 0.22) was significantly higher than that of normal nasopharynx tissues (0.14 +/- 0.07, F = 27.94, P < 0.05). The relative expression level of CXCR4 protein in NPC (1.58 +/- 0.59) was significantly higher than that of normal nasopharynx tissues (0.51 +/- 0.22, F = 17.75, P < 0.05). The high expression levels of CXCR4 mRNA and protein in NPC were closely related to clinical stage, cervical lymph node metastasis and cancer cell differentiation (P < 0.05). SDF-1alpha protein was strongly expressed in normal superior deep cervical lymph nodes, bone marrow, lung and liver (2.35 +/- 0.67), but absent or very poor expression in inferior deep cervical lymph nodes, kidney and colon tissues (0.68 +/- 0.23), and the differences between them were statistically significant (t = 10.13, P < 0.01).</p><p><b>CONCLUSION</b>CXCR4 is closely correlated to metastasis of nasopharyngeal carcinoma. CXCR4/SDF-1alpha axis may play an important role in organ-specific metastasis of NPC.</p>

Influence of suppression of Epstein-Barr Virus-encoded latent membrane protein 1 by rAAV vector mediated RNA interference on metastatic ability of nasopharyngeal cancer cells in vivo / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the effect of Epstein-Barr virus (EBV) encoded latent membrane protein 1 (LMP-1) on the tumor growth and metastasis of nasopharyngeal carcinoma (NPC) cells in vivo and its possible mechanism.</p><p><b>METHODS</b>To construct two recombinant adeno-associated virus (rAAV): rAAV-shRNA-LMP-1 and rAAV-EGFP (enhanced green fluorescent protein). Multiplicity of infection (MOI) was confirmed by using different titre of rAAV-EGFP to transfect a NPC cell line, C666-1. Then C666-1 cells were transfected by rAAV-shRNA-LMP-1 at MOI titre and the inhibiting efficiency of target gene's expression was confirmed by reverse transcription polymerase chain reaction (RT-PCR). C666-1 cells treated by RNAi on LMP-1 in vitro was directly inoculated into the liver via laparotomy under direct vision to establish the animal model of NPC xenograft in liver and lung metastasis from the liver. The biological effect and its mechanism after "gene silencing" of LMP-1 on NPC cells tumorigenesis and metastasis were observed by the primary intrahepatic tumor formation and lung metastasis and the expression of matrix metalloproteinases-9 (MMP-9) revealed by immunohistochemistry.</p><p><b>RESULTS</b>The transfection efficiency was higher than 95% with 5 x 10(4) virus genome (v.g)/cell with rAAV-EGFP. The expression of target gene was inhibited more than 90%, assessed by RT-PCR after transfection with rAAV-shRNA-LMP-1 at a dose of 5 x 10(4) v.g/cell. The primary tumor volume implanted in the liver of rAAV-shRNA-LMP-1 treatment was (0.2527 +/- 0.1152) cm3, with no significant difference in comparrison with rAAV-EGFP control group [(0.2533 +/- 0.0754) cm3, P>0.05]. But the rate of intrahepatic tumor formation was 50.0% and the rate of lung metastasis was 33.3% of the rAAV-shRNA-LMP-1 group, significantly lower than those in the rAAV-EGFP group (P<0.05). The survival time (15.50 +/- 2.47) d of rAAV-shRNA-LMP-1 group was significantly longer than that in the rAAV-EGFP group [(11.50 +/- 1.22) d, P<0.05]. Immunohistochemistry indicated suppression of LMP-1 by rAAV-shRNA-LMP-1 can down-regulate the expression of MMP-9 significantly.</p><p><b>CONCLUSION</b>The expression of LMP-1 can be suppressed effectively by rAAV mediated RNA interference. The suppression of LMP-1 expression has no effect on cell growth but can inhibit the metastasis in vivo, probably through down-regulating the expression of MMP-9.</p>

Effect of rAAV-mediated RNA interference against EB virus-encoded latent membrane protein 1 on nasopharyngeal carcinoma cell proliferation and metastasis in nude mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the role of Epstein-Barr virus (EBV)-encoded latent membrane protein 1 (LMP-1) in nasopharyngeal carcinoma (NPC) cell growth and metastasis in vivo by recombinant adeno-associated virus (rAAV)-mediated RNA interference (RNAi).</p><p><b>METHODS</b>Specific small hairpin RNA (shRNA) targeting EBV-LMP-1 gene was designed and synthesized to construct two rAAV vectors rAAV-shRNA-LMP-1 and rAAV-EGFP. The multiplicity of infection (MOI) was confirmed using different titers of rAAV-EGFP to transfect the NPC cell line C666-1. The C666-1 cells were transfected by rAAV-shRNA-LMP-1 at the optimal MOI titer and the inhibition efficiency of the target gene expression was determined with RT-PCR. The C666-1 cells with RNAi of LMP-1 gene were injected into nude mouse liver via laparotomy to establish the animal model of hepatic and lung metastases of NPC cells. The metastases of the C666-1 cells in the liver and lungs were observed to assess the effect of LMP-1 gene silencing on the tumorigenic and metastatic potentials of the cells in vivo.</p><p><b>RESULTS</b>The transfection efficiency of 5 x 10(4) virus genome/cell rAAV-EGFP exceeded 95%. The expression of the target gene was suppressed by over 90% as shown by RT-PCR after transfection with rAAV-shRNA-LMP-1 at 5 x 10(4) virus genome/cell. Animal experiments showed that compared with rAAV-EGFP, rAAV-shRNA-LMP-1 transfection did not reduce the primary tumor volume implanted into the liver, but significantly inhibited the intrahepatic and lung metastases of the NPC cells.</p><p><b>CONCLUSION</b>LMP-1 expression can be suppressed effectively by rAAV-mediated RNAi, and LMP-1 suppression does not obviously affect the tumor cell growth but can inhibit their metastasis in vivo.</p>

Pepsin immunoassay in the sputum for detection of laryngopharyngeal reflux / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study whether the presence of gastric pepsin in the sputum might be used as a reliable criteria of laryngopharyngeal reflux.</p><p><b>METHODS</b>Fifty-six patients with the symptoms of laryngopharyngitis and fifteen healthy people were recruited. Fifty-six patients were divided into laryngopharyngeal reflux group and chronic laryngitis group by the reflux symptom index (RSI), by the reflux finding score (RFS) and by their treating experiment taking omeprazole 20 mg bid for 2 weeks. Sputum in all three groups was obtained in the morning. Pepsin in the sputum was measured by enzyme-linked immunoadsorbent assay.</p><p><b>RESULTS</b>The positive rate of pepsin in sputum among LPR group, chronic laryngopharyngitis group and normal group were 93.8% (30/32), 75.0% (18/24), 20.0% (3/15) respectively, and the median concentration of pepsin were 5.3 [1.3; 53.4] ng/ml, 0.8 [0.1; 17.2] ng/ml, 0.0[0.0;0.0] ng/ml (H = 23.29, P = 0.000). Compared with co-diagnosis as gold standard, the sensitivity of RSI, RFS treating experiment and the pepsin immunoassay was 59.4%, 84.4%, 81.3% and 93.8%, and the specificity of those was 87.2%, 61.5%, 95.8% and 46.2% respectively.</p><p><b>CONCLUSIONS</b>Detection of pepsin in sputum by immunoassay might provide a high sensitive, noninvasive method for laryngopharyngeal reflux.</p>

Experiment of inhibiting growth and metastasis of xenograft transplanted human nasopharyngeal carcinoma in nude mice by interferon alpha-1b / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To observe the growth and metastasis effect of interferon alpha-1b (IFN-alpha-1b) on nasopharyngeal carcinoma cell line CNE-2 in xenografted model of mice liver. Comparing rAAV-mediated IFN-alpha-1b gene therapy with the IFN-alpha-1b protein therapy.</p><p><b>METHODS</b>The xenografted model of liver nasopharyngeal carcinoma was established by injecting the human nasopharyngeal carcinoma cell line CNE-2 under liver capsule of nude mice. Forty nude mice were randomly divided into four groups by means of random number table method, with ten mice in each one. (Group A: rAAV-IFN-alpha-1b, Group B: IFN-alpha-1b; Group C: rAAV-EGFP; Group D: PBS). After 24 hours, A, C and D group was injected with rAAV-IFN-alpha-1b encoding human IFN-alpha-1b, rAAV-EGFP and phosphate buffer saline via tail vein injection. After 5 days, mice in group B was injected with human IFN-alpha-1b protein once per two days. Three weeks later five nude mice were sacrificed and then observed their liver tumor formation and pulmonary metastasis. Tumor size was measured and tumor inhibition ratios was calculated, and apoptotic index (AI) was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL). The contents of human IFN-alpha contained in peripheral blood and mice IL-12 was determined by high performance liquid chromatography chip techniques. And another five mice were randomly chosen for the observation of surviving study.</p><p><b>RESULTS</b>After human nasopharyngeal carcinoma implants in nude mice liver 3 weeks, the average volume of A group (0.114 +/- 0.116) cm3 and B group (0.422 +/- 0.137) cm3 were significantly lower than that of C group (2.476 +/- 0.637) cm3 and D group (2.677 +/- 0.704) cm3 (F = 38.536, P < 0.01). Compared with D group, the restrained percentage of tumor in group A was 95.74% and group B was 84.24%. The percentage of lung metastases in group A, B, C and D were 0.0%, 0.0%, 40.0%, 60.0% respectively. The apoptotic index increased significantly in group A (21. 88 +/- 3.29)% and group B (19.85 +/- 1.96)% versus group C (4.37 +/- 0.50)% and group D (3.40 +/- 1.05)% (F = 120.964, P < 0.01). The average content of human interferon-alpha in serum increased significantly in group A (101.50 +/- 11.33) pg/ml and group B (91.55 +/- 9.80) pg/ml versus group C (23.06 +/- 4.36) pg/ml and group D (16.93 +/- 9.96) pg/ml (F = 69.128, P < 0.01). The average content of IL-12 increased significantly in group A (80.36 +/- 13.35) pg/ml and group B (51.15 +/- 9.72) pg/ml versus group C (19.44 +/- 7.03) pg/ml and group D (14.49 +/- 4.21) pg/ml (F = 57.116, P < 0.01). The survival time of tumor bearing mice in group A (55.80 +/- 2.77) d and group B (48.20 +/- 2.39) d was significantly longer than group C (35.40 +/- 2.61) d and group D (36.80 +/- 1.92) d (chi2 = 25.623, P < 0.01).</p><p><b>CONCLUSIONS</b>IFN-alpha-1b can inhibit the growth and metastasis of nasopharyngeal carcinoma cell line CNE-2 in xenografted model of mice liver. rAAV-mediated IFN-alpha-1b gene therapy indicated more effect than the IFN-alpha-1b protein therapy by comparing content of human IFN-alpha in serum and the survival time of tumor bearing mice.</p>

Effect of estrogen replacement therapy in the early phase on atherosclerosis and PAI-1 in ovariectomized cholesterol-fed rabbit / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of estrogen replacement therapy (ERT) in the early phase on the atherosclerosis and the level of plasminogen activator inhibitor-1(PAI-1).@*METHODS@#Twenty-eight rabbits were randomly assigned to 4 groups: Group A, sham operation (n=7); Group B, ovariectomized without estradiol (n=7); Group C, ovariectomized with low-dose estradiol (n=7); and Group D, ovariectomized with high-dose estradiol (n=7). All rabbits were given 1% cholesterol diet for 12 weeks. Levels of blood lipid, estradiol, and PAI-1 were measured before the operation and at the end of the 4th and 12th weeks. Twelve weeks later, we took the aortas for pathological analysis and calculated the areas of atherosclerotic plaque.@*RESULTS@#After 12 weeks, the estradiol level of Group B was significantly lower than that of Group A, and that of Group D was obviously higher than Group A. There was no significant difference between Group C and A. The concentrations of total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) in Group B significantly increased compared with Group A (P<0.01). The levels of TC and LDL-C of Group C and D were significantly lower than those of Group A. Whereas the concentrations of triglyceride (TG) and high density lipoprotein cholesterol (HDL-C) in Group B were lower than those of Groups A, C and D (P<0.01). In contrast to Groups A, C and D, the level of PAI-1 was significantly higher in the Group B (P<0.01), without significant differences among Groups A, C and D. The area of atherosclerotic lesion of aorta in Group B was significantly bigger than that of Group A, C and D. The areas of aortic atherosclerotic plaque in Group C and D were obviously smaller than those of Group A (P<0.01).@*CONCLUSION@#Transdermal estrogen replacement therapy in the early phase can improve the metabolism of the serum lipids, reduce the level of PAI-1, and probably provide the protective effect on the atheroma formation.