Objective To evaluate the prognosis for patients with squamous cell carcinoma of cervical lymph node metastases from an unknown primary site who had different clinical stages and were irradiated to different fields. Methods The clinical date of 60 patients with cervical lymph node metastatic squamous cell carcinoma from an unknown primary site(no including patients with supraclavicular node metastases) were retrospectively analyzed. Results The overall 5-year survival rates(OSR) was 68.5%, the OSR of patients with N 1,N 2 and N 3 stage were 100%, 68.0% and 40.9% respectively(? 2=7.29,P=0.026). The OSR of patients with one lateral neck, whole neck and large fields irradiation were 66.5%, 74.5% and 54.6% respectively(? 2=1.38,P =0.501). The Cox proportional hazard model showed that clinical stages of cervical lymph node had an significant effect in patients survival(P=0.032). The 5-year local control rates(LCR) 65.6%, the LCR of patients with N 1,N 2 and N 3 stage were 100%, 63.2% and 34.6% respectively(? 2=5.51, P=0.064). The LCR of patients with one lateral neck, whole neck and large fields irradiation were 87.6%, 51.0% and 72.7% respectively(? 2=2.55, P =0.279). The 5-year subsequent appearance rates of occult primary cancer(SAR) was 21.2%, the SAR of patients irradiated by small(one lateral neck or whole neck) and large fields were 23.3% and 12.5% respectively(? 2=0.52, P =0.469). Conclusions The clinical stages of cervical lymph node is an important prognostic factor for survival and the local control rates has a decreasing trend as the stage increases. The subsequent appearance rates of occult primary cancer of patients irradiated by small fields has a higher trend than its of patients irradiated by large fields, but the control rates and survival rates have no significant difference in different fields irradiated.

Objective To investigate the correlation of HIF-1α and hypoxia in keloids fibroblasts, and to investigate the mechanism that hypoxia promotes abnormal scarring by HIF-1α pathway. Methods Keloid fibroblasts cultured in vitro were placed in an incubator with different O2 concentrations. After 24 h, the keloid fibroblasts were collected for further study. Western blotting was performed to detect the expression of HIF-1α in the keloid fibroblasts. Results Relative amounts of HIF-1α in keloid fibroblasts cultured under O2 concentrations at 20 %, 10 %, 5 % and 1 % were 0. 007 ±0. 006, 0. 133 ±0. 006, 0. 537±0. 015 and 0. 903±0. 021, respectively. It indicated that hypoxia could increase the expression of HIF-lα in keloid fibroblasts. Conclusions Hypoxia can induce the expression of HIF-1α in fibroblasts of keloids. Moreover, there still is a positive relation between hypoxia and the expression of HIF-1α. Therefore, a close relationship exists between abnormal scarring and HIF-1α pathway by hypoxia.

AIM: To observe the effect of microRNA-16 (miR-16) on the megakaryocytic differentiation of K562 cells, and to explore the potential mechanism.METHODS:miR-16 was over-expressed or silenced by transfection with miR-16 mimics or inhibitor in K562 cells.The level of miR-16 was detected by real-time PCR.The expression of CD41, CD42b and CD61, as megakaryocytic differentiation markers, was detected by flow cytometry.The effect of miR-16 on the expression of myeloblastosis oncogene ( MYB) was measured by Western blotting, and flow cytometry was performed to confirm whether the effect of miR-16 on expression of CD41, CD42b and CD61 was mediated by MYB.RESULTS:Transfection with miR-16 mimics dramatically elevated the level of miR-16 and the expression of CD41, CD42b and CD61 in the K562 cells.Transfection with miR-16 inhibitor decreased the level of miR-16 and the expression of CD41, CD42b and CD61 in the K562 cells (P<0.05).The expression of MYB was regulated by miR-16, and MYB silencing reversed the regulation of CD41, CD42b and CD61 induced by miR-16.CONCLUSION:miR-16 regulates the megakaryocytic dif-ferentiation of K562 cells by targeting MYB.

Objective To explore the clinical value of gonadotropin-releasing hormone analogue (GnRH-a) by analyzing the efficiency of GnRH-a treatment for moderate and severe endometriosis after laparoscopic conservative operations.Methods Two hundred and sixty-five females who accepted laparoscopic conservative operation were enrolled in this study,and they were confirmed by pathology for ovarian endometriosis cyst,in phase Ⅲ andⅣ,47 patients had fertility desire.According to whether use the drug and auxiliary species the postoperative cases were divided into 5 groups:102 cases without endocrinal therapy(untreated group),GnRH-a group of 3 months in 64 cases,GnRH-a group of 6 months in 31 cases;gestrinone group of 3 months in 38 cases,and gestrinone group of 6 months in 30 cases.All patients were followed up for 2 years,observed drug clinical effects,remission rate,improvement rate,recurrence rate,pregnancy outcomes and side effects.Results The remission rate in GnRH-a group of 6 months and gestrinone group of 6 months was significantly higher than that in untreated group [83.9％ (26/31),73.3％ (22/30) vs.52.9％(54/102)],the remission rate in GnRH-a group of 6 months was significantly higher than that GnRH-a group of 3 months [64.1％ (41/64)],there was significant difference (P ＜ 0.05).The recurrence rate in GnRH-a group of 3 months,GnRH-a group of 6 months,gestrinone group of 6 months was significantly lower than that in untreated group [12.5％ (8/64),9.7％ (3/31),10.0％ (3/30) vs.27.5％ (28/102)] (P ＜ 0.05).The recurrence rate in gestrinone group of 6 months was significantly lower than that in gestrinone group of 3 months (P ＜ 0.05).The recurrence time of GnRH-a group of 3 months,GnRH-a group of 6 months,gestrinone group of 6 months was significantly longer than that in untreated group [(14.05 ± 1.97),(16.76 ± 1.53),(16.12 ±2.15)months vs.(12.85 ± 1.80)months] (P ＜0.05).The recurrence time of gestrinone group of 3 months and untreated group had no significant difference(P ＞ 0.05).The pregnancy rate of GnRH-a group and gestrinone group was higher than that in untreated group,but there was no significant difference (P ＞ 0.05).The rate of menopausal symptom in GnRH-a group was higher than that in gestrinone group [50.5％ (48/95) vs.16.2％ (11/68)].The rate of abnormal bleeding in vagina,acne and side-effect of aminotransferase in GnRH-a group were significantly lower than those in gestrinone group [1.1％(1/95) vs.7.4％(5/68),0 vs.8.8％(6/68),0 vs.5.9％(4/68)](P＜ 0.05).Conclusions Laparoscopic conservative operations combined with GnRH-a on treatment of moderate and severe ovarian endometriosis can significantly improve remission rate,decrease recurrence rate.GnRH-a or gestrinone for 6 months,the pregnancy rate increases.In addition the side reactions of the two medicine have statistic difference,which of GnRH-a is lower.

Objective To define the correlation between mutation of DNA-LigaseⅣ gene and radiosensitivity.Methods Nasopharyngeal squamous carcinoma cell line (CNE), lung adenocarcinoma cell line (SPC-A1) and breast adenocarcinoma cell line(MCF-7) after irradiation were assessed with specific biological parameters. Polymerase chain reaction, cloning and sequencing techniques were used to determine the sequence of DNA-LigaseⅣ gene in these three cell lines. Then the impact of homologous change and mutation on hydrophicity-hydrophobicity of genic products was analyzed.Results The surviving fractions derived from irradiation were different with more radiosensitivity in the CNE cell line than in the others. In three cell lines, the homology of LigaseⅣ gene were: 99.95%,99.99%,99.98%, respectively. Some mutations including transversion and transition were detected and led to alterations in the hydrophicity-hydrophobicity function of products. Higher radiosensitivity of CNE was associated with amino-acid substitutions: 313aa His→Arg,538aa Gly→Arg,579aa Lys→Arg and 585aa Asn→Ser.Conclusion These results suggest that LigaseⅣp play an important role in the ligation of DNA double strand breaks and certain mutations bring about changes in radiosensitivity.

Objective To investigate the change in mRNA expression of hypoxia inducible factor 1? HIF 1? and vascular endothelial growth factor (VEGF) genes in the highly differentiated nasopharyngeal squamous carcinoma (CNE) cell line during hypoxia reoxygenation.Methods CNE cells were cultured in normoxic, continuous 8 hour hypoxia ,16 hour hypoxia and 16 hour hypoxia 4 hour reoxygenation conditions respectively. Expressions of HIF 1? and VEGF genes were assessed by the reverse transcription polymerase chain reaction (RT PCR) technique. Results The mRNA expression of HIF 1? gene was kept constant in the different culture conditions,whereas the mRNA expression of VEGF gene increased significantly after 8 or 16 hour hypoxia and decreased after reoxygenation for 4 hours. Conclusions Hypoxia is able to induce the mRNA expression of VEGF gene in the CNE cell, but is unable to do so in HIF 1? mRNA expression. These findings suggest that VEGF upregulation induced by hypoxia be independent HIF 1a transcription.

AIM: To investigate a new method for identification of traditional Chinese medicine injection by FTIR. METHODS: For the steadiness of spectra, the factors of effecting spectrum's information quality were all investigated scientifically over the experiment procedures and instrumental setting, such as the preparation of samples, resolution ratio, scanning times, repeating scanning times, etc. The traditional Chinese medicine injections were used as the analytical samples such as Radix Isatidis, Rhizoma Chuanxiong, Flos Carthami, Radix Astragali and Herba Houttuyniae. RESULTS: Although all these original spectrums were similar at a certain degree, the FTIR combined with computer aided analysis, such as the cluster analysis and derivative spectrometry comparability calculation could be used to identify these injections. CONCLUSION: The method of identification by FTIR is non destructive testing, cheap, clean, fast, simple and convenient. The result indicates this method is suitable for establishing identification database of traditional Chinese medicine injections.

Objective:To study the effect of rotational errors on the positioning accuracy (PA) and to assess whether correcting rotation in patients with head-neck tumors in radiotherapy or not.Methods:The image information of 34 patients with head-neck tumors treated at Zhongnan Hospital of Wuhan University between August 2019 and January 2020 was collected. Mega-voltage computed tomography (MVCT) images of each patient were taken before radiotherapy, and were registered with planned kilo-voltage computed tomography (KVCT) images by two registration methods. All information was divided into control group (translation only) and intervention group (translation and rotation) according to different registration methods, there were 144 fractioned registered images for each group, respectively. The position errors of the two registration methods were recorded and compared. Data were carried out with Wilcoxon signed rank test and Spearman rank correlation.Results:Translational errors of the control group and the intervention group were 0.10 (5.35) mm and 0.00 (5.78) mm in right-left direction, and there was a statistically significant difference ( Z=-2.675, P=0.007); 0.75 (2.78) mm and 0.60 (2.68) mm in superior-inferior direction, and there was a statistically significant difference ( Z=-2.819, P=0.005); 0.10 (0.90) mm and 0.20 (1.28) mm in anterio-posterior direction, and there was a statistically significant difference ( Z=-3.984, P<0.001). Rotational errors of the intervention group were -0.20 (0.60)°, 0.35 (2.00)°, 0.00 (0.98)° in pitch, roll, yaw, respectively. The distribute of 3D vector corrected frequency for two groups was positively skewed. The corrected cumulative frequency (CCF) varied with 3D vector, 3D vector was 8.0 mm, and 19 F and 16 F fractioned treatments of the control group and the intervention group were not corrected, respectively; 3D vector was between 8.0-13.5 mm, the corrected tendency of the intervention group was slower and fractioned treatment was completed later. The analytical results of Spearman rank correlation showed that rotational errors in pitch were negatively correlated with translational errors of the control group in superior-inferior direction ( r=-0.182, P=0.029) and the intervention group in anterio-osterior direction ( r=-0.484, P<0.001); rotational errors in roll were negatively correlated with translational errors of the intervention group in right-left direction ( r=-0.334, P<0.001); rotational errors in yaw which were positively correlated with translational errors of the intervention group in right-left direction ( r=0.370, P<0.001) were negatively correlated with translational errors of the control group in superior-inferior direction ( r=-0.171, P=0.040) and the same was true for the intervention group ( r=-0.203, P=0.015); total angles were positively correlated and negatively correlated with translational errors of the control group in superior-inferior direction ( r=0.246, P=0.003) and anterio-posterior direction ( r=-0.188, P=0.024), and positively correlated with 3D vector of the control group ( r=0.198, P=0.017), total angles were positively correlated with translational errors of the intervention group in superior-inferior direction ( r=0.170, P=0.041) and with 3D vector of the intervention group ( r=0.239, P=0.004); there were no correlations between rotational errors and the other translational errors (all P>0.05). Conclusion:Although the corrected rotation increases translational errors in anterio-posterior direction and 3D vector, it improves PA for head-neck tumors in radiotherapy. When rotational errors are not corrected, rotational offsets are present with corrected translation to decrease its effect on PA.

Objective To investigate the spatial organization of neurons, blood vessel and astrocytes at different time of reperfusion after focal cerebral ischemia in rats. Methods 24 Sprague-Dawley rats were randomly divided into sham group (n=8), reperfusion 1 day group and reperfusion 2 weeks group. The latter 2 groups were occluded the middle cerebral arteries for an hour and reperfused. All the rats were injected with gelatin ink. The expressions of glial fibrillary acid protein (GFAP) and neuronal nuclear antigen (NeuN) in the brain were observed with immunohistochemistry. Results The vessels located mainly in cortex and nucleus. Most of astrocytes apophysis connected with vessels and neurons. Compared to the sham group, the expression of GFAP increased significantly in ischemic side, and the expression of NeuN decreased 1 day and 2 weeks after ischemia-reperfusion. The vessels decreased in the ischemic side 1 day after cerebral ischemia-reperfusion, and then increased 2 weeks later. Conclusion The organization of neurovascular unit after cerebral ischemia-reperfusion has been observed.

Objective To establish and evaluate 3 kinds of minimally-invasive valve implantation model in vivo.Methods A novel tissue engineered heart valve(TEHV) manufactured by branched polyethylene glycol cross-linked acellular porcine valve and a minimally-invasive valve implantation system according to the design of Corevalve revalving system were adopted.After anesthesia,18 adult male goats were randomly divided into 3 groups: the ulrasound-directed orthotropic group (group A,n =6),angiography-directed orthotropic group (group B,n =6) and direct-released heterotopic group (group C,n =6),and all received minimally-invasive valve implantation orthotropically or heterotopically.4 weeks later,the valvular function was evaluated by CTA and/or echocardiography.Results All 3 kinds of caprine model were successfully constructed.The operation success rate of each group was A: 66.7％,B: 50.0％ and C: 100.0％,respectively(multiple x2 analysis,group A and B P ＞0.05; group A and C,group B and C,P ＜0.05).The operation-time of each group was A: (79 ± 18) min,B:(61 ±23) min,C: (45 ± 15) min(one-way ANOVA,P ＜0.05).The survival rate at4 weeks was A: 100％,B: 100％ and C: 83.3％ (multiple x2 analysis,P ＞ 0.05).Echocardiography and CTA proved the short-term function of implanted TEHV was satisfactory.Conclusion All 3 kinds of caprine valve implantation model can be established without cardiopulmonary bypass and blood transfusion.The devices and equipments required in group A is relatively simple,but the procedure cost longer time for it is hard to determine the right position by ultrasound.The application of angiography made the positioning much easier in group B while the procedure had to be performed in specific operation room with angiographic apparatus.Group C did rely on neither special equipments nor complex operation,but the valve leaflets cannot work normally,so this model was only suitable for testing in vivo characteristics such as biocompatiblities.