1.TRAP1 gene silencing effect on biological properties of CD24-CD44+human laryngeal squamous cell carcinoma stem cells
Jing SU ; Haitao XUE ; Junhai TIAN ; Jihua ZHANG
Chinese Journal of Tissue Engineering Research 2017;38(5):663-668
BACKGROUND:Studies have indicated that the abnormal expression of tumor necrosis factorreceptor-associated protein 1 (TRAP1) is closely related to the occurrence and development of a variety of tumors. Therefore, targeted inhibition of TRAP1 expression has become an important target for the treatment or intervention of tumor growth. OBJECTIVE:To explore the effect of the TRAP1 gene silencing on the proliferation and apoptosis of human laryngeal squamous cel carcinoma stem cel s. METHODS:CD24-CD44-human laryngeal squamous cel carcinoma stem cel s were isolated by flow cytometry. Interfering RNA (siRNA) sequences for smal molecule TRAP1 gene was designed and transferred into human laryngeal cancer stem cel s by LipofectamineTM 2000. Flow cytometry, MTT assay, cel clone formation assay and TUNEL apoptosis assay were used to evaluate the effect of silencing TRAP1 gene on the proliferation and apoptosis of CD24-CD44+laryngeal cancer stem cel s. RESULTS AND CONCLUSION:Compared with CD24+CD44-cel s, CD24-CD44+cel s upregulated OCT4, SOX2, NANOG and TRAP1 expression levels (P<0.05). However, the expression of TRAP1 protein in human laryngeal squamous cel carcinoma was significantly decreased after RNA interference (P<0.05). The growth rate of TRAP1 gene silenced human laryngeal squamous cel carcinoma was significantly reduced (P<0.05), the cel arrest was in the G0/G1 phase, the number of cel s in the S phase was decreased (P<0.05), and there was no significant change in the M phase. TRAP1 gene silencing significantly inhibited the proliferation of human laryngeal squamous cel carcinoma stem cel s (P<0.05). Compared to the non-transfected cel s, the TRAP1 gene silencing significantly reduced the clone formation ability of transfected human laryngeal squamous cel carcinoma stem cel s (P<0.05), and TRAP1 gene silenced-human laryngeal squamous cel carcinoma stem cel s were more easy to trigger apoptosis by upregulating BAD and BAX expression levels (P<0.05). Overal , our experimental results indicate that the specific interference of TRAP1 gene expression could inhibit the proliferation and promote apoptosis of human laryngeal squamous cel carcinoma stem cel s.
2.Clinical observation of modified uvulopalatopharyn goplasty combined with coblation in the treatment of obstructive sleep apnea patients
Dongmei SONG ; Baoshan WANG ; Changchun SUN ; Haitao XUE ; Junhai TIAN ; Lijuan MU
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(03):-
OBJECTIVE On the basis of the different shapes of upper airway(UA)obstruction and collapse characteristic of obstructive sleep apnea hypopnea syndrome(OSAHS), the effect of modified uvulopalatopharyngoplasty(UPPP)combined with coblation in OSAHS operations on patients with AHI≥20,which could be done selectively to patients with various or multiple UA obstructive sites, was explored. METHODS 30 patients of OSAHS were diagnosed and analysed by polysomnography(PSG)to have AHIs≥20. Based on the shapes of their UAs classified according to the degree of the shortened lateral and radial vectors,the UPPP was modified in two ways: either the lateral vector or the radial vector was amplified, or both could be amplified with coblation. All patients were analyzed by questionnaires(PSG)Muller's maneuver and oral cavity measurement pre-and 6 months post-operation. RESULTS 86.7 % of the patients showed a decrease of at least 25 % in AHI . Velopalatal insufficiency never occurred. CONCLUSION The modified-UPPP combined with coblation can be used selectively on patients based on their UA shapes.
3.RNA interference targeting inhibition of TRAP1 suppresses cell growth and promotes apoptosis in CD133+CD44+ laryngeal carcinoma stem cells
Haitao XUE ; Jing SU ; Shuai CHEN ; Chunju CHEN ; Jihua ZHANG ; Junhai TIAN ; Kaifeng DONG
Chinese Journal of Tissue Engineering Research 2017;21(17):2672-2677
BACKGROUND: Tumor necrosis factor-associated protein 1 (TRAP1) is a heat-shock protein 90-related mitochondrial chaperone. Accumulative evidence has demonstrated that TRAP1 overexpression is closely related to carcinogenesis. However, the exact function and mechanism of TRAP1 in the occurrence of laryngeal carcinoma remains unclear. OBJECTIVE: To investigate whether RNA interference can inhibit TRAP1 overexpression and to explore its effects on growth and apoptosis of CD133+CD44+ laryngeal carcinoma stem cells. METHODS: CD133+CD44+ laryngeal carcinoma stem cells were sorted from human laryngeal carcinoma Hep-2 cellsusing immunomagnetic beads. The shRNA sequence of TRAP1 was designed and synthesized and CD133+CD44+ laryngeal carcinoma stem cells were transfected with LipofectamineTM 2000. Cell counting kit-8 assay, colony formation assay and flow cytometry were used to investigate the effects of interference of TRAP1 expression on growth and apoptosis of CD133+CD44+ laryngeal carcinoma stem cells. Spectrophotometric method was used to detect the activity of caspase-3, -8 and -9. RESULTS AND CONCLUSION: TRAP1 mRNA and protein expression levels were significantly decreased in TRAP1 shRNA-transfected CD133+CD44+ laryngeal carcinoma stem cells (P < 0.01). Compared with the blank control and negative control groups, the growth and colony formation of CD133+CD44+ laryngeal carcinoma stem cells were significantly inhibited in the TRAP1 shRNA-transfected group (P < 0.05). Apoptosis of CD133+CD44+ laryngeal carcinoma stem cells was significantly inhibited in the TRAP1 shRNA-transfected group as compared with the blank control and negative control groups (P < 0.05). TRAP1 shRNA-mediated cell apoptosis was associated with the activation of caspase-3, -8 and -9. These results suggest that RNA interference targeting inhibition of TRAP1 suppresses cell growth but promotes apoptosis in CD133+CD44+ aryngeal carcinoma stem cells. TRAP1 is likely to be a gene target for treatment of laryngeal carcinoma.
4.Screening of chemokine receptor CCR4 antagonists by capillary zone electrophoresis
Zhe SUN ; Linjie TIAN ; Qian LIN ; Xiaomei LING ; Junhai XIAO ; Ying WANG
Journal of Pharmaceutical Analysis 2011;01(4):264-269
CC chemokine receptor 4 (CCR4) is a kind of G-protein-coupled receptor, which plays a pivotal role in allergic inflammation. The interaction between 2-(2-(4-chloro-phenyl)-5-{[(naphthalen-1-ylmethyl)-carbamoyl]-methyl}-4-oxo-thiazolidin-3-yl)-N-(3-morpholin-4-yl-propyl)-acetamide (S009) and the N-terminal extracellular tail (ML40) of CCR4 has been validated to be high affinity by capillary zone electrophoresis (CZE). The S009 is a known CCR4 antagonist. Now, a series of new thiourea derivatives have been synthesized. Compared with positive control S009, they were screened using ML40 as target by CZE to find some new drugs for allergic inflammation diseases. The synthesized compounds XJH-5, XJH-4, XJH-17 and XJH-1 displayed the interaction with ML40, but XJH-9, XJH-10, XJH-11, XJH-12, XJH-13, XJH-14, XJH-3, XJH-8, XJH-6, XJH-7, XJH-15, XJH-16 and XJH-2 did not bind to ML40.Both qualification and quantification characterizations of the binding were determined. The affinity of the four compounds was valued by the binding constant, which was similar with the results of chemotactic experiments. The established CEZ method is capable of sensitive and fast screening for a series of lactam analogs in the drug discovery for allergic inflammation diseases.
5.The evaluation of virtual endoscopy and fiberoptic endoscopy in the diagnosis of obstructive sleep apnea syndrome.
Zhiming LIU ; Qingyun REN ; Baoshan WANG ; Haitao XUE ; Junhai TIAN ; Junlu ZHAO
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2008;22(6):257-259
OBJECTIVE:
The utility of virtual endoscopy is compared to fiberoptic endoscopy and was also investigated with respect to accuracy of diagnosis and reproduction of images in patients with obstructive sleep apnea hypopnea syndrome (OSAHS).
METHOD:
Twenty-one patients with OSAHS were examined by helical spiral CT axial images and fiberoptic endoscopy. The helical spiral CT axial data was reconstructed using a VE software program. The results were compared to the fiberoptic endoscopic findings done by the otolaryngologists. All the patients were examined both in the sleeping and nonsleeping status. The dimensions of soft palate, uvula, lingua and epiglottis region were evaluated.
RESULT:
The results both in the virtual endoscopy evaluation and fiberoptic endoscopy was statistically significant difference in all of region. In the palate region, there was statistically significant difference in the left-and-right dimension, but no difference in the fore-and-aft dimension both in the sleeping and nonsleeping status.
CONCLUSION
The dimensions of upper airway were more difference between in the sleeping and nonsleeping status. The change of dimension was more in the left-and-right; the fiberoptic endoscopy has more diagnostic significance in evaluation of dynamic movement of the upper airway; virtual endoscopy evaluation of the upper airway was accurate in assessing stenosis width and length; virtual endoscopy added more information about anatomic structure and pathological change of the upper airway.
Adult
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Endoscopy
;
methods
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Female
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Humans
;
Laryngoscopy
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Male
;
Middle Aged
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Sleep Apnea, Obstructive
;
diagnosis
;
diagnostic imaging
;
Tomography, Spiral Computed
;
Tomography, X-Ray Computed
6.Surveillance study of irrigating fluid absorption and bleeding in transurethral resection of the prostate
Junhai MA ; Ning FAN ; Chengzhou LU ; Huiming GUI ; Yunxin ZHANG ; Gongjin WU ; Hong CHANG ; Ze QIN ; Zhongjin YUE ; Jun MI ; Li YANG ; Junqiang TIAN ; Panfeng SHANG ; Zhilong DONG ; Zhiping WANG
Chinese Journal of Urology 2019;40(1):37-41
Objective To investigate the effect of the endoscopic surveillance system in irrigating fluid absorption and bleeding during transurethral resection of the prostate.Methods In vitro trials,we simulated the fluid absorption and bleeding in the operation by using self-developed endoscopic surveillance system from January 2013 to June 2013.Continuous irrigation of 5 % mannitol solution,we extracted 5 times irrigating fluid (each time 100 ml and a total of 500 ml) in the process of irrigation and recorded absorption measurements of every time extraction rinses.At the same time,we dripped human whole blood 5 times(each time 5 ml and a total of 25 ml) in the process of irrigation and recorded the bleeding measurements.The above process was repeated three times to detect the accuracy and consistency of the endoscopic surveillance system.In clinical trials,50 cases of BPH were monitored in surgery and the biochemical index,hemodynamics,irrigating fluid absorption and bleeding were compared from October 2016 to April 2017.The included criteria contained as follow:the age of patients should be more than 50 years.The transabdominal ultrasound showed that the volume of prostate should be more than 60 ml.The maximal uroflowmetry should be less than 15ml/s.The IPSS scores should be more than 8.Based on the operative time,two groups (<60 min and ≥ 60 min) were classified.Results We developed the endoscopic surveillance system which is original in the world.In vitro trials,the average irrigating fluid were (100.60 ± 2.07) ml,(201.00±3.39) ml,(302.00±4.67) ml,(403.60±4.39) ml and (502.40 ±7.57) ml;and the average bleeding were (5.06 ± 0.11) ml,(10.10 ± 0.16) ml,(15.04 ± 0.15) ml,(20.06 ± 0.11) ml and (25.10 ± 0.16) ml.No significant difference was observed in all groups (P > 0.05).In clinical trials,we compared some preoperative and postoperative indexes.The average blood oxygen saturation were (94.46 ± 2.49) % and (92.39 ± 2.77) % (P < 0.01),the average Serum sodium ion concentration were (141.05 ± 2.52) mmol/L and (138.06 ± 4.27) mmol/L(P < 0.01),the average HGB were (143.50 ± 13.43) g/L and (137.04 ± 14.25) g/L(P < 0.01).The average HCT were (42.05 ± 4.09) % and (137.04 ± 14.25) % (P < 0.01).The average HR were (77.9 ± 7.6) beats per minute and (77.93 ± 6.93) beats per minute (P>0.05).The MAP were (90.32 ± 9.75) mmHg and (91.07±8.96)mmHg(P>0.05).The average serum potassium ion concentration were (4.13 ± 0.53) mmol/L and (4.09 ± 0.37) mmol/L (P > 0.05).The average irrigating fluid absorption of the group less than 60 minutes and the group equal or more than 60 minutes were (401.83 ± 279.23) ml and (885.25 ± 367.68) ml (P < 0.01).The average blood loss were (64.10 ±47.47) ml and (158.40 ± 65.22) ml(P <0.01).The preoperative and postoperative hemodynamic,blood biochemical and hematology showed difference in our trials.Irrigating fluid absorption and blood loss were positively associated with operation time.Conclusions The endoscopic surveillance system was safety and accuracy.It can offer real-time monitoring data and alarm mechanism for the surgeons that possibly improve operation safety.