OBJECTIVE To evaluate the effect of AG490 on the invasion of Hep2 laryngeal carcinoma cells, and to investigate the role of STAT3 signaling pathway in human laryngeal carcinoma invasion.METHODS Janus kinase inhibitor AG490 was used to inhibit the activation of STAT3 signaling pathway of Hep2 cells.Hep2 cell invasion was detected by quantitative Matrigel invasion assay.The expression of p-STAT3 was measured by flow cytometry. RESULTS The activation of STAT3 signaling pathway and the invasion of laryngeal carcinomas Hep2 cell line were suppressed by AG490.The expression of p-STAT3 and the invasion cell number of Hep2 cell line in the group treated by AG490 for 24h decreased compared with that in control group(P

Objective To compare effect of chemotherapy agent DDP to MACS in sorting cancer stemcells (CSC) of laryngeal carcinoma cell line Hep-2. Methods CD133 magnetic beads were applied to sort Hep-2 cells. Different dosages of DDP were used to treat Hep-2 cells for 48 hours. Enrichment rate of CD133+ cells by MACS and after DDP treatment was detected by Flow Cytometer (FCM). Morphologic change was observed under inverse-phase microscope. Results FCM showed that the sorting rate of CD133+ cells through MACS was 64.33 %, while after DDP treatment for 48 hours, the rate of CD133+ cells was enriched significantly in each dosage of DDP, with the maximal rate was 50.7 %, in the dosage of 4 μg/ml. There was a significantly difference between MACS and each of DDP group (P <0.01). Cells treated with DDP were abnormal in morphology. Conclusion MACS and DDP sorting has respective advantages in enriching CSC in Hep-2 cell lines.

OBJECTIVE To evaluate the inhibitory effect of HPSE AS-ODN on the invasiveness of human Hep-2 cell lines. METHODS HPSE AS- ODN which was complementary with initiation codon region of HPSE mRNA was designed and synthesized. After embedded by cation lipofectin, it was transfected into Hep-2 cells of human laryngocarcinoma. The expression of HPSE protein and HPSE mRNA in Hep-2 cell lines were detected by flow cytometry and RT- PCR. Meanwhile Matrigel invasive assay was used to measure the inhibitory effect of HPSE AS-ODN on the invasiveness of human Hep-2 cell lines. RESULTS The HPSE protein and HPSE mRNA expression and invasiveness of human Hep-2 cells treated with AS- ODN of different concentrations were significantly decreased as the AS-ODN concentration increasing. There was a significantly difference between control group and each group of AS-ODN respectively (P

Background and purpose:Proteasome inhibitors constitute a novel class of antitumor agents that has a complex mechanism of action.Previous studies have confirmed that proteasome inhibitor MG-132 can significantly inhibit Hep-2 cell growth and induce cell apoptosis in a manner that is dependent on dosage and time.But it also induced p-STAT3 protein expression.The aim of this study was to explore whether the STAT3 gene can,by transfecting short hair pin RNA(shRNA),enhance the anti-tumor effect of MG-132 on human laryngeal carcinoma cells.Methods:Hep-2 cells were plated into 96-well and 6-well plates and incubated overnight.Then,they were treated with MG-132 alone and combined with pshSTAT3.Their cell growth was detected by MTT assay,and apoptosis was examined with flow cytometry.The protein expression of p-STAT3 was detected by Western blotting.Results:MTT assay showed that a combined group inhibited the proliferation of Hep-2 cells compared to the MG-132 group and pshSTAT3 group(P＜0.01).Flow cytometry showed that apoptosis of the combined group was significantly higher than the MG-132 group and pshSTAT3 group (P＜0.01).Western blotting showed that the p-STAT3 protein expression up-regulation was observed in the MG-132 group,whereas down-regulation was expressed in the combined group and pshSTAT3 group.Conclusion:The shRNA targeting STAT3 gene can prevent the up-regulation of p-STAT3 protein following a MG-132 treatment thereby significantly enhancing the anti-tunlor effect of the protease inhibitor,MG-132,on human laryngeal carcinoma cells.

Objective To investigate the effect of milk basic protein (MBP) on bone metabolism in normal and ovariectomized (Ovx) rats. Method Forty-eight female Sprague-Dawley rats were ovariectomized and another 12 rats received sham operation (Sham). After 10 d recovery period, the Ovx rats were randomly divided into 4 groups: control, low-dose, medium-dose, and high-dose MBP group. Another 44 normal female rats without ovariectomy were also divided into 4 groups as above. The MBP dosages for each group were respectively 0, 10, 20, 30 mg/kg bw. All rats were i.g. administered for 90 d. Bone mineral density (BMD) of the femur (at proximal end, middle of diaphysis, and distal end) was measured by dual-energy X-ray absorptiometry in vivo. The amounts of calcium, magnesium and phosphorus were analyzed by ICP-AES. Results BMD at distal end of femur was significantly higher in normal low-dose group than in normal control group while no significant effect was observed in Ovx MBP groups. As for the amounts of calcium, magnesium and phosphorus, there were no significant differences among normal experimental groups and also among Ovx experimental groups. However, some variations in the level of those minerals were observed. Conclusion MBP at 10 mg/kg bw significantly elevated BMD at femoral distal end in normal rats, while no similar effect was observed in Ovx rats. Besides its influence on bone minerals, there may be another mechanism involved in its effect on bone metabolism.

Objective: To sammarize the methodology of the treatment of alveolar cleft by autogenous cancellous bone graft obtained with mini incision(ABGM).Methods: 22 cases of alveolar cleft received ABGM by one operator. The cleft area was restored with autogenous cancellous bone from iliac crest by cylinder osteotomes for extraction of bone transplants(COEBT) with mini incision. All the patients were follow up for at least six months postoperatively. The result of the bone grafting was evaluated on the radiographs. Results: ①The overall survival rate of ABGM was 95.8%,and clinical successful rate was 91.7%;②The successful rate of ABGM in group of patients above 18 year old, or with complete cleft palate or bilateral cleft lip and palate was lower than that in other groups.③The wound for extracting autogenous cancellous bone from iliac crest by COEBT with mini incision was smaller than that by osteotomy. Conclusion: ABGM with COEBT is an effective way in the treatment of alveolar cleft. The successful rate of ABGM is related with the age and cleft style of the patients.

Objective To obtain more detailed microsurgical anatomical data and multi-slice helical CT scanning of cochlear aqueduct for the translabyrithine approach. Methods Twenty cadaver heads(40 sides)fixed with formalin were dissected under a microscope ,and cochlear aqueducts were found and measured. Five cadaver heads(10 sides) were scanned by multi-slice helical CT. Results The width of external aperture of cochlear aqueduct was (2.64?0.82)mm. The length of cochlear aqueduct was (8.40?1.35)mm. The distance between cochlear aqueduct and cranial nerves Ⅸ, Ⅹ, Ⅺ, and that between the inferior petrosal sinus was(1.08?0.36 )mm,(2.71?0.63)mm,(4.52?1.25)mm, (1.14?0.46)mm, respectively. The distance between external aperture of cochlear aqueduct and the inferior margin of internal acoustic meatus aperture was (5.53?0.93)mm. The otic capsule segment of cochlear aqueduct could not be demonstrated on the coronal section. The petrosal portion could be demonstrated by both coronal and axial scannings. Conclusion The axial scanning was useful for detecting cochlear aqueduct. Cochlear aqueduct could be regarded as one of the landmarks in translabyrithine approach for cerebellopontile angle operation.

Objective Study the relationship between type A influenza virus genetic variation with survival selective pressure, help for the finding of possible vaccine conserved antigen target. Methods Select seven strains of same HA (Hemagglutinin) serotype, regional and isolation time closely related type A influenza virus with full HA gene coding sequence; use Blast2 program to calculate the parameter of nucleotide conservative, amino acid conservative, mutation ratio of codon 3rd (over) non 3rd locus, survival selective pressure indicator of these virus strains; analysis the parameters relationship with survival selective pressure indicator. Results Nucleotide conservative of HA gene is significantly lower than that of other genes; amino acid conservative of HA gene is similar with NS gene, all lower than that of other genes; genetic variation of HA gene is under survival selective pressure; selective pressure toward NS, PB1 gene is relatively lower than that toward NA, NP gene; survival selective pressure indicator is strongly correlated with mutation frequency upon codon 3rd locus. Conclusions Genetic variation of influenza virus is determined both by mechanism of relax replication model and survival selective pressure; genetic conservative of each gene is different, NA, NP gene could be selected as possible vaccine target for their relative high conservative, HA gene possesses medium genetic conservative with prospective of finding more conserved epitope region within its full sequence, NS, PB1 are not recommended as vaccine candidate for their relative low genetic conservative.

OBJECTIVE: To investigate the influence of signal transducer and activator of transcription 3 (Stat3) and hypoxia-inducible factor-lα (HIF-1α) on the resistance effect of laryngeal squamous cell carcinoma to radiation therapy and chemotherapy under the hypoxia circumstances. METHOD: Western blot was used to test the expression of p-Stat3 and HIF-1α in the Hep-2 cells under the hypoxia conditions. MTT assay was used to test the proliferation of Hep-2 cells after radiation therapy and chemotherapy; the Hep-2 cells were suppressed expression of Stat3 and/or HIF-1α. RESULT: (1) AG490 induced significant proliferation inhibition on Hep-2 cells and Hep-2HIF-1α-/- cells in vitro underthe hypoxia environments (P < 0.05); (2) Suppressing expression of Stat3 reduced the expression of HIF-1α protein (P < 0.05); (3) Combined inhibition of Stat3 and HIF-1α enhanced radio- and chemo-sensitivity in laryngeal squamous carcinoma cells under hypoxia. CONCLUSION Combined inhibition of Stat3 and HIF-1α can further enhance radio- and chemo-sensitivity in laryngeal squamous carcinoma cells under hypixia compare than inhibiting Stat3 or HIF-1α alone. Effectively blocking of HIF-1α pathway and suppressing the expression of Stat3, would be an effective method to enhance radio- and chemo-sensitivity in laryngeal squamous carcinoma cells, which provides a new thought to reduce the resistance to treatment.
Blotting, Western ; Carcinoma, Squamous Cell ; Cell Hypoxia ; Cell Line, Tumor ; Humans ; Hypoxia ; Hypoxia-Inducible Factor 1, alpha Subunit ; Laryngeal Neoplasms ; STAT3 Transcription Factor

Objective:To study the expression of CD137L in laryngeal carcinoma, and to analyze its clinicopathological significance.Method:The expression of CD137L in 50 laryngeal carcinoma specimens and 9 normal laryngeal mucous tissues were detected by immunohistochemical staining.Result:The positive CD137L staining were found in all 50 cases of laryngeal carcinomas (100%), while its staining were negative in normal laryngeal mucous. There was significant difference between two groups(P<0.01). The positive ratio of CD137L staining had no relationship with the factors such as age, sex and tumor site, while it had significant correlation with the pathological stage,T stage and lymph node metastasis.Conclusion:The expression of CD137L might play an important role in the development of laryngeal carcinomas.