[Comparison of quantitative detection of BCR::ABL1 p210 transcript levels: a multicenter study].

Objective: To assess the capability of seven reference medical laboratories to detect BCR::ABL1 p210 transcription levels and to compare the results among those laboratories. Methods: The interlaboratory comparison was carried out in two stages. The samples were prepared by the reference laboratory. The quantitative values of BCR::ABL1 p210 of the comparison samples covered 0.001%-0.01%, 0.01%-0.1%, 0.1%-1%, 1%-10% and>10% in each stage. Real-time quantitative PCR (RT-PCR) and dPCR (digital PCR) were used to examine the samples. The conversion factor (CF) was calculated and validated for each laboratory. Results: In the RT-PCR comparison, one laboratory was failed to detect BCR::ABL1 p210 in fourteen samples at the first stage. The results of the other six laboratories were qualified with the bias <±1.2 folds (-0.133-0.338) and 95% limits of agreement within ±5 folds (upper limit 0.147-0.785, lower limit -0.770--0.109), and the corresponding CF values were calculated and validated. In the dPCR comparison, one laboratory did not report results at the second stage. The results of the other six laboratories were qualified with the bias <±1.2 folds (-0.026-0.267) and 95% limits of agreement within±5 folds (upper limit 0.084-0.991, lower limit -0.669--0.135), and the corresponding CF values were calculated and validated. The samples with BCR::ABL1 p210 quantitative values of 0.01%-0.1%, 0.1%-1%, 1%-10% and >10% could be detected by both RT-PCR and qPCR. When the quantitative value of BCR::ABL1 p210 was 0.001%-0.01%, the detection rate of dPCR was higher than that of RT-PCR (85.56% vs. 68.00%). Conclusions: A good consistency is present among various laboratories. The quantitative value of BCR::ABL1 p210 is comparable among laboratories as shown by the CF value conversion. For quantitative detection of BCR::ABL1 p210 deep molecular reaction, dPCR has a higher positive detection rate and more advantages than RT-PCR. To ensure the accuracy and reproducibility of the BCR::ABL1 p210 test, it is imperative for every laboratory to enhance their daily quality control practices.

[Clinical outcomes of endovascular therapy in acute stroke patients with anterior circulation tandem occlusions due to atherosclerosis and dissection].

Objective: To compare the clinical outcomes of endovascular therapy in acute stroke patients with anterior circulation tandem occlusions caused by atherosclerosis or dissection. Methods: A retrospective cohort study. A total of 98 patients with anterior circulation tandem lesions undergoing endovascular therapy in the Wuhan NO.1 Hospital (March 2016 to March 2022) were analyzed. Median age was 64(55,71) years old, and 82.7% (81/98 cases) were males. According to the lesion etiology, the patients were divided into atherosclerosis and dissection groups. The differences in clinical outcomes between the two groups were investigated, including favorable 90-day functional outcome (modified Rankin Scale score of 0-2), successful reperfusion (modified Thrombolysis in Cerebrovascular Infarction score of 2b-3), symptomatic intracranial hemorrhage, stroke-associated pneumonia, 90-day all-cause mortality, and average hospitalization days. Logistic regression analysis was used to adjust for potential confounders affecting functional outcomes in both groups, and to determine odds ratios and 95% confidence intervals. Results: Seventy-one patients were grouped into the atherosclerotic cause and 27 into the dissection cause cohorts. The rate of favorable 90-day functional outcome was 43.7% (31/71 cases) in the atherosclerosis group versus 55.6% (15/27 cases) in the dissection group (adjusted odds ratio=1.339; 95% confidence interval, 0.374-4.798; P=0.654). No significant differences were found in other clinical outcomes between the two groups (all P>0.05). Conclusion: The clinical prognosis of patients with tandem lesions caused by atherosclerotic stenosis or artery dissection was similar after endovascular therapy. Future studies are still needed to verify our results.

[Experimental study on the therapeutic effect and mechanism of erlotinib on non-proliferative diabetic retinopathy].

Objective: To investigate the therapeutic effect and mechanism of erlotinib, an epidermal growth factor receptor (EGFR) inhibitor, on non-proliferative diabetic retinopathy (NPDR). Methods: An experimental research was conducted. Human retinal Müller cells (RMC) were MIO-M1 cells from Moorfields Ophthalmology Hospital and the Institute of Ophthalmology at London University College. MIO-M1 cells were divided into normal, hypertonic, high glucose, high glucose+dimethyl sulfoxide (DMSO), high glucose+erlotinib 0.5 mmol/L, high glucose+erlotinib 1 mmol/L, and high glucose+erlotinib 2 mmol/L groups using a random number table method. Detection of the effect of erlotinib on the proliferation of MIO-M1 cells under high glucose conditions was performed by 5-ethynyl-2'-deoxyuridine (EdU) method. Western blotting (WB) was used to detect the effect of erlotinib on the activation markers of glial fibrillary acidic protein (GFAP) and glutamine synthetase (GS) protein levels in MIO-M1 cells under high glucose conditions. WB was used to detect the effect of erlotinib on the protein levels of nerve growth factor receptor (p75NTR), vimentin, and cell retinol binding protein (CRALBP) in RMC under high glucose conditions. MIO-M1 cells were divided into normal group, high glucose group, high glucose+DMSO group, and high glucose+erlotinib (1 mmol/L) group using random number table method. The effect of erlotinib on EGFR nuclear translocation under high glucose conditions was detected by cell immunofluorescence staining. Immunoprecipitation was used to detect the effect of erlotinib on the interaction between EGFR and transcription intermediate factor 2 (TIF2) in MIO-M1 cells under high glucose conditions. MIO-M1 cells were randomly divided into normal group, high glucose group, high glucose+DMSO group, high glucose+Myc-DDK empty body group, high glucose+erlotinib group, high glucose+erlotinib+human doublet protein group, high glucose+erlotinib+TIF2 plasmid group, and high glucose+erlotinib+human doublet protein+TIF2 plasmid group. Cell immunofluorescence staining was used to detect the effect of erlotinib on the binding of EGFR and TIF2 in MIO-M1 cells under high glucose conditions through the EGFR/TIF2 axis. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the regulatory effect of EGFR and TIF2 binding on cyclin D1 transcription in MIO-M1 cells under high glucose conditions. The mouse model of diabetes retinopathy (DR) was constructed and divided into normal group, DR group, DR+DMSO group, DR+erlotinib 0.25 mg·kg-1·d-1 group, DR+erlotinib 0.5 mg·kg-1·d-1 group and DR+erlotinib 1 mg·kg-1·d-1 group. 25 mice in total, 5 in each group. Tissue immunofluorescence staining was used to detect the expression of RMC activation marker GFAP. The FITC-dextran injection experiment was used to detect the effect of erlotinib on retinal vascular leakage in a murine DR model. Results: Compared with the normal group (32.4%±3.0%), the proportion of EdU positive cells in RMC in the high glucose group (59.2%±3.8%) increased (P<0.001). Compared with the high glucose group (59.2%±3.8%), the proportion of EdU positive cells in the high glucose+1 mmol/L erlotinib group (37.6%±4.4%) decreased (P<0.001). Compared with the normal group, the expression of GFAP in RMC in the high glucose group increased (1 in the normal group, 2.27±0.11 in the high glucose group, P<0.001), while the expression of GS decreased (1 in the normal group, 0.32±0.03 in the high glucose group, P<0.001). 1 mmol/L erlotinib treatment reduced the expression of GFAP in RMC under high glucose conditions (1.32±0.13 and 2.27±0.11, respectively; P<0.001), and increased the expression of GS (0.71±0.06 and 0.32±0.03, respectively; P<0.001). The colocalization of EGFR and DAPI in RMC of the high glucose+1 mmol/L erlotinib group was lower than that of the high glucose group (52.2%±4.1% and 76.4%±5.7%, respectively; P<0.001). The expression of TIF2 or EGFR both increased while using EGF or TIF2 antibodies to precipitate TIF2 or EGFR under high glucose conditions compared to the normal group (1 in the normal group, 2.27±0.20 in the high glucose group, 2.17±0.21 in the EGFR, all P<0.05). And the expression of TIF2 (1.38±0.10) or EGFR (1.32±0.13) in the high glucose+erlotinib group was lower than that in the high glucose group (2.27±0.20) and the high glucose group (2.17±0.21) (all P<0.05). The colocalization of EGFR and TIF2 (17.2%±3.9%) and the mRNA level of Cyclin D1 (1.32±0.16) in the RMC of the high glucose+erlotinib group were lower than those in the high glucose group (54.6%±3.7% of EGFR and TIF2 colocalization ratio, 2.58±0.19 of Cyclin D1 mRNA level,all P<0.05). The high glucose+erlotinib+AREG (EGFR agonist) group, high glucose+erlotinib+Myc DDK-TIF2 plasmid group and high sugar+erlotinib+AREG+Myc-DDK-TIF2 plasmid group EGFR colocalization with TIF2 (colocalization ratios 24.1%±1.9%, 26.0%±2.3%, 35.3%±2.5%) and TIF2 mRNA levels (1.71±0.16, 1.72±0.18, 2.20±0.18). Compared with the high glucose+erlotinib group, The increases were statistically significant (all P<0.05). Compared to the normal group, the expression of GFAP in mouse retina tissue was increased in the DR group (1 in the normal group, 3.07±0.19 in the DR group, P<0.001), and 0.5 mg·kg-1·d-1 erlotinib (1.73±0.30) significantly reduced the expression of GFAP in the retina of DR group mice (P<0.05). Compared to the normal group (3.97±0.47), the DR group (23.13±2.15) showed an increase in fluorescein leakage, while the DR+erlotinib group (11.66±1.45) showed a significant decrease in leakage compared to the DR group (all P<0.05). Conclusions: Erlotinib inhibits the proliferation and activation of RMC induced by high glucose, inhibits the entry of EGFR into the nucleus, inhibits the binding of EGFR to TIF2 in RMC, and reduces the transcription of Cyclin D1 in RMC by inhibiting the interaction between EGFR and TIF2. At the same time, erlotinib inhibits the proliferation and activation of RMC in the mouse DR model, ameliorating retinal vascular leakage in mice. These results suggest that erlotinib inhibits the activation and proliferation of RMC by downregulating the EGFR/TIF2/Cyclin D1 pathway under high glucose conditions, thereby alleviating the progression of NPDR.

[Arf6 regulates endometriotic epithelial-mesenchymal transition and mitochondrial distribution].

Objective: To investigate the role of adenosine diphosphate ribosylation factor 6 (Arf6) in the pathogenesis of endometriosis. Methods: Endometrial tissues were sampled from women who were hospitalized in the Affiliated Hospital of Medical School of Ningbo University and Ningbo Women and Children's Hospital from November 2020 to May 2021 with endometriosis (n=44, endometriosis group) and without endometriosis (n=17, control group). The expression of Arf6 protein in the endometrial tissues was detected by western blot. Endometrial epithelial cells from both groups were primary cultured and the distribution of intracellular mitochondria was detected by immunofluorescence. The expression of Arf6 protein was down-regulated by small interference RNA (siRNA), the distribution of mitochondria in cells with decreased Arf6 protein expression was observed, and the expression of mitochondria-related proteins development and differentiation enhancing factor 1 (DDEF1, also called AMAP1), reactive oxygen species 1 (ROS1) and epithelial-mesenchymal transition (EMT)-related proteins E-cadherin, vimentin were detected. Transwell assay was used to detect the changes in the migration ability of the cells. Results: Compared with the control group, ectopic endometrial tissue of endometriosis group showed high expression of Arf6 protein (0.174±0.019 vs 0.423±0.033; t=29.630, P<0.01); and in ectopic endometrial epithelial cells, mitochondria were distributed near the edge of the cell membrane. While Arf6 expression was down-regulated by siRNA, the distribution of mitochondria in ectopic cells returned to natural, close to the control level. In addition, the expression levels of AMAP1 and ROS1 in ectopic cells after Arf6 protein knockdown were significantly decreased. Transwell assay results indicated that knockdown of Arf6 could reduce the migration ability of ectopic epithelial cells [migration cell count: (34.3±7.5) cells]; and immunofluorescence verified low expression of E-cadherin but high expression of vimentin in ectopic epithelial cells, whereas knockdown of Arf6 protein E-cadherin expression increased but vimentin expression decreased. Conclusions: High expression of Arf6 protein in ectopic endometrial epithelial cells leads to the distribution of mitochondria tending to membrane marginalization, while inducing EMT, which are involved in the mechanism of endoheterosis pathogenesis.

[Quantification of optical coherence tomographic angiography images in type 2 diabetic patients].

Objective: To quantify optical coherence tomographic angiography (OCTA) images and to explore potential clinical values of each parameter in diabetic patients. Methods: A case-control and cross-sectional study of diabetic (129 eyes) and healthy (32 eyes) subjects was conducted. 3-mm OCTA scan, fundus photography and best corrected visual acuity measurement were performed. Image-pro plus was used to calculate microvascular and foveal avascular indices in nonsegmented and segmented OCTA images. Intraclass correlation coefficient and relative standard deviation were used to examine the reliability, reproducibility and accuracy. Correlation of each parameter was calculated, and so was the area under the receiver operating characteristic curve. Results: Vessel density (VD) of diabetic patients measured in the nonsegmented retinal layer (RL) and superficial retinal layer (SRL) was 49.146%±6.097%, 44.038%±5.641%, respectively, significantly lower than healthy subjects 52.212%±6.250%, 46.698%±5.417% (t=2.534, 2.405; P=0.012, 0.017), and vessel length (VL) was (19.905±2.285)mm, (17.596±2.149)mm, (14.479±2.091)mm in the three layers, which was significantly shorter than that in the healthy control group (21.037±2.185)mm, (18.739±1.994)mm, (15.343±2.266)mm (t=2.529, 2.731, 2.059; P=0.012, 0.007, 0.041). Compared with non-diabetes, foveal avascular zone (FAZ) in diabetes was (0.441±0.167)mm(2), larger compared to health eyes 0.352±0.109mm(2) (t=-2.831, P=0.005). FAZ demonstrated a negative correlation with VD and VL in RL or SRL(r=-0.227, -0.338; P<0.05) (r=-0.241, -0.332; P<0.05), while best corrected visual acuity showed a negative correlation with VL in each layer (r=-0.225, -0.201, -0.250; P=0.01, 0.022, 0.004), and VD in DRL (r=-0.197, P=0.026). All parameters showed high reproducibility between graders(intraclass correlation coefficient>0.965), while relative standard deviation was greater than 3.049%. AR had the highest area under the receiver operating characteristic curve (0.737), compared with AI (0.724). Conclusions: Our study demonstrates good reliability, reproducibility and accuracy of quantification of OCTA images and reveals changes of VD, VL and FAZ in diabetes, which may occur earlier than vision impairment. AR and AL may have great values in prompting early diabetic retinopathy in diabetic patients. (Chin J Ophthalmol, 2019, 55:273-279).

ANO1 in intramuscular interstitial cells of Cajal plays a key role in the generation of slow waves and tone in the internal anal sphincter.

KEY POINTS: The internal anal sphincter develops tone important for maintaining high anal pressure and continence. Controversy exists regarding the mechanisms underlying tone development. We examined the hypothesis that tone depends upon electrical slow waves (SWs) initiated in intramuscular interstitial cells of Cajal (ICC-IM) by activation of Ca2+ -activated Cl- channels (ANO1, encoded by Ano1) and voltage-dependent L-type Ca2+ channels (CavL , encoded by Cacna1c). Measurement of membrane potential and contraction indicated that ANO1 and CavL have a central role in SW generation, phasic contractions and tone, independent of stretch. ANO1 expression was examined in wildtype and Ano1/+egfp mice with immunohistochemical techniques. Ano1 and Cacna1c expression levels were examined by quantitative PCR in fluorescence-activated cell sorting. ICC-IM were the predominant cell type expressing ANO1 and the most likely candidate for SW generation. SWs in ICC-IM are proposed to conduct to smooth muscle where Ca2+ entry via CavL results in phasic activity that sums to produce tone. ABSTRACT: The mechanism underlying tone generation in the internal anal sphincter (IAS) is controversial. We examined the hypothesis that tone depends upon generation of electrical slow waves (SWs) initiated in intramuscular interstitial cells of Cajal (ICC-IM) by activation of Ca2+ -activated Cl- channels (encoded by Ano1) and voltage-dependent L-type Ca2+ channels (encoded by Cacna1c). Phasic contractions and tone in the IAS were nearly abolished by ANO1 and CavL antagonists. ANO1 antagonists also abolished SWs as well as transient depolarizations that persisted after addition of CavL antagonists. Tone development in the IAS did not require stretch of muscles, and the sensitivity of contraction to ANO1 antagonists was the same in stretched versus un-stretched muscles. ANO1 expression was examined in wildtype and Ano1/+egfp mice with immunohistochemical techniques. Dual labelling revealed that ANO1 expression could be resolved in ICC but not smooth muscle cells (SMCs) in the IAS and rectum. Ano1, Cacna1c and Kit gene expression were the same in extracts of IAS and rectum muscles. In IAS cells isolated with fluorescence-activated cell sorting, Ano1 expression was 26.5-fold greater in ICC than in SMCs while Cacna1c expression was only 2-fold greater in SMCs than in ICC. These data support a central role for ANO1 and CavL in the generation of SWs and tone in the IAS. ICC-IM are the probable cellular candidate for ANO1 currents and SW generation. We propose that ANO1 and CavL collaborate to generate SWs in ICC-IM followed by conduction to adjacent SMCs where phasic calcium entry through CavL sums to produce tone.

Macrodeformation Twins in Single-Crystal Aluminum.

Deformation twinning in pure aluminum has been considered to be a unique property of nanostructured aluminum. A lingering mystery is whether deformation twinning occurs in coarse-grained or single-crystal aluminum at scales beyond nanotwins. Here, we present the first experimental demonstration of macrodeformation twins in single-crystal aluminum formed under an ultrahigh strain rate (∼10^{6} s^{-1}) and large shear strain (200%) via dynamic equal channel angular pressing. Large-scale molecular dynamics simulations suggest that the frustration of subsonic dislocation motion leads to transonic deformation twinning. Deformation twinning is rooted in the rate dependences of dislocation motion and twinning, which are coupled, complementary processes during severe plastic deformation under ultrahigh strain rates.

blaKPC and rmtB on a single plasmid in Enterobacter amnigenus and Klebsiella pneumoniae isolates from the same patient.

Enterobacter amnigenus (EA76) and Klebsiella pneumoniae (KP76) isolates with multidrug-resistant (MDR) patterns were identified from the same patient in the neurosurgery department of our hospital. An outbreak of MDR K. pneumoniae had also occurred in this department. To characterize the resistance mechanism and molecular epidemiology of these isolates, sequential experiments including antimicrobial susceptibility testing, polymerase chain reaction (PCR), plasmid analysis, pulsed field gel electrophoresis (PFGE), and multilocus sequence typing (MLST) were performed. EA76 and KP76 were resistant to all of the antibiotics tested, except colistin and tigecycline. blaKPC-2, blaTEM-1, blaSHV-12, blaCTX-M-3, blaCTX-M-14, and rmtB genes were identified in both isolates, with blaKPC-2, blaTEM-1, blaCTX-M-14, and rmtB being co-carried on one plasmid in each isolate. Further analysis showed different restriction patterns between the two KPC-carrying plasmids. Of the 11 carbapenem-resistant isolates found in the outbreak, all were resistant to all of the ß-lactams tested, with 63.64% (7/11) also exhibiting resistance to aminoglycosides and 72.73% (8/11) exhibiting resistance to quinolones. PCR analysis and molecular typing of the 11 K. pneumoniae strains revealed that the seven aminoglycoside-resistant isolates shared the same antibiotic-resistant gene pattern and identical or one-band-difference PFGE profiles relative to KP76. In addition, all of the eight aminoglycoside-resistant isolates, including KP76, belonged to the national epidemic clone ST11. The overall results indicate the emergence of E. amnigenus and outbreak of ST11 K. pneumoniae, with both co-harboring blaKPC and rmtB genes on a single plasmid in our neurosurgery wards.

[The surgical effectiveness of montgomery thyroplasty implant on patients with unilateral vocal fold paralysis].

Objective: To investigate the efficacy of type I thyroplasty with Montgomery prosthesis implantation for the treatment of unilateral vocal fold paralysis. Methods: From May 2015 to March 2019, 46 patients (24 males, 22 females, with age range of 23-77) with unilateral vocal fold paralysis underwent thyroplasty with Montgomery prosthesis implantation in the Department of Otorhinolaryngology Head and Neck Surgery in both the First Affiliated Hospital of Navy Medical University and Guangdong Provincial People's Hospital. The assessment methods included GRBAS auditory perception assessment, acoustic analysis such as Jitter, Shimmer, NHR and maximum phonation time (MPT). Results: Postoperative videostroboscopy observed the displacement of paralyzed vocal fold to the midline in 44 cases as well as significantly reduced glottic fissures during phonation. In the other 2 cases, glottic fissure did not reduce significantly. Compared with preoperative data, the scores of all parameters in GRBAS auditory perception assessment were lower except the parameter S, and the acoustic analysis parameters (jitter, shimmer, NHR) were smaller, and MPT was longer. All the difference was statistically significant (P<0.001). Revision surgery was performed in 2 patients with poor results. No serious complications occurred in all the cases. Conclusions: For the patients with unilateral vocal fold paralysis who are not suitable for the laryngeal reinnervation surgery due to old age or long course of denervation, thyroplasty with Montgomery prosthesis implantation can effectively improve the voice of patients with high safety,which is worthy of promotion.

[Clinical analysis of selective laryngeal reinnervation using upper root of phrenic nerve and hypoglossal nerve branch in the treatment of bilateral vocal fold paralysis].

Objective: To evaluate the airway and voice quality improvement in patients with bilateral vocal fold paralysis (BVFP) who underwent selective laryngeal reinnervation surgery. Methods: From January 2012 to December 2016, a retrospective study was conducted in 39 patients with BVFP who underwent selective laryngeal reinnervation surgery in Department of Otorhinolaryngology Head and Neck Surgery, the First Affiliated Hospital of Navy Medical University. All patients were examined by videostroboscopy, vocal function assessment, laryngeal electromyography and pulmonary function test before and after the surgery, and followed up for at least 2 years to evaluate the efficacy and safety of the surgery.Wilcoxon signed rank test was used to analyze the G score and VHI-10 score data. Paired t-test was used to analyze acoustic parameters, MPT values and pulmonary function parameters. Results: Postoperative infection and hemorrhage occurred in one patient separately.Videostroboscopic videos showed that at 4-8 months postoperatively, vocal folds in 35 patients achieved moderate or severe abduction during inspiration, 2 patients only achieved mild abduction, 2 patients showed no abduction,while all patients achieved adduction in bilateral vocal cords during phonation. The recovery rate of moderate-to-severe abduction was 89.7% (35/39), and these patients were decannulated successfully. At 12 months after operation, G score and VHI-10 score were significantly lower than those before operation (P<0.05), and the acoustic parameters jitter, shimmer, HNR and MPT were significantly improved (P<0.05). Most of the parameters of the pulmonary function test at 3 months postoperatively returned to the normal reference level, while the maximum inspiratory pressure (PImax) at 12 months after operation was still slightly lower than the normal level, but it was significantly improved compared with preoperative value (P<0.05). The EMG data at 12 months postoperatively showed full interference potentials in 37 patients in bilateral posterior cricoarytenoid muscles during inspiration, and full interference potentials in bilateralthyroarytenoid muscles during phonation. Obvious misdirected regeneration electric activitieswere found in two of them. Potentials in posterior cricoarytenoid muscle were weak in 2 cases with poor abduction. During long-term follow-up, only one case showed decreased abduction, but did not affect respiratory function. Conclusions: The selective laryngeal reinnervation procedure applied in the present study can restore physiological motion of vocal cords. The success rate was high, the curative effect was stable, and the complications were rare. It is worth of promotion.

[Clinical observation of three-dimensional printing donor tooth model in peri-operative period of autotransplantation of tooth].

Objective: To prepare a three-dimensional (3D) printing donor tooth model and to observe its application in the peri-operative period. Methods: In part one, 192 cases (2017.9-2019.8) from Department of Oral & Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University ï¼»107 males and 85 females, age (34.2±10.7) yearsï¼½ which need autotransplantation of teeth (ATT) were collected. Whether the donor teeth can be completely extracted was predicted through clinical and imaging examination (first prediction). The second prediction was supplemented by the three-dimensional printing model of the donor teeth. Each of the prediction was compared with the actual results and the coincidence rate was calculated. In part two, 64 cases (2017.9-2019.8) from Department of Oral & Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University ï¼»28 males, 36 females, age (30.2±8.3) yearsï¼½ which need ATT were randomly divided into the model group and the donor group. The time of alveolar fossa preparation, time of donor tooth in vitro, times of trial implantation and time of pulptomy and root canal location were recorded respectively. Results: In part one, the coincidence rate between the second prediction and the actual results ï¼»97.4%(187/192)ï¼½ was significantly higher than that of the first prediction ï¼»93.2%(179/192)ï¼½ (P<0.05). In part two, the preparation time of the alveolar fossa in the maxillary and mandibular were (18.8±4.6) and (22.7±3.4) min, the time of the teeth in vitro were (3.0±0.6) and (2.1±0.6) min, the times of trial implantation were (1.3±0.8) and (1.0±0.9), and the time of pulpotomy and root canal location were (4.3±0.6) and (4.0±0.5) min. All values in the model groups were better than those in the donor group (P<0.05). Conclusions: The 3D printing model is accurate. It can be used in autogenous tooth transplantation to shorten the preparation time of alveolar fossa and time of donor tooth in vitro, and reduce the times of trial implantation of donor teeth, and to help to improve the prediction accuracy of complete extraction of donor teeth and the time of pulpotomy and root canal location.

Correlation between adiponectin polymorphism and atherosclerotic plaque compositions under intravascular ultrasound (IVUS).

OBJECTIVE: To discuss the correlation between polymorphism of rs266729 (-11377C/G, Cytosine/Guanine) (adiponectin promoter) site and atherosclerotic plaque compositions as well as related indicators under intravascular ultrasound (IVUS). PATIENTS AND METHODS: 76 patients with coronary heart disease from December 2014 to December 2016 were enrolled. The PCR-RFLP method was used to analyze the adiponectin gene polymorphism in rs266729 site. All the objects were divided into CC type group (n=26), CG type group (n=23), and GG type group (n=27) according to the results of polymorphism. The amount of lesions and length of lesion in the vessel were determined according to the images of coronary angiography. The indicators from each group, including minimum external elastic membrane area, the smallest lumen area, the patch area, the patch load, the lipid pool area, the lipid pool/plaque area, the fiber cap thickness, the reconstruction index, the positive reconstruction, the negative reconstruction and the patch character were measured according to the IVUS results. RESULTS: The baseline data from distinct the gene types showed no significant difference. The results of quantitative IVUS plaque analysis indicated a statistical difference of factors such as plaque area, plaque burden, lipid pool area, lipid pool/plaque area, and remodeling index between the CC and GG types (p<0.05). The levels of aminopeptidase N (APN), tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1ß), fasting serum insulin (FIN), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) among diverse groups presented statistical difference (p<0.05). Of note, the analysis results of IVUS qualitative components of plaque showed that soft plaque in CC group was 42.3% (11/26), which was significantly lower than GG group 11.1% (3/27) (p<0.05). The vascular remodeling ratio in CC group 26.9% (7/26) was also significantly decreased compared to that in GG group 66.7% (18/27) (p<0.05). The tubular and diffuse ratio in CC groups according to the comparison of diseased vessel, count, length of the lesion were 34.6% (9/26) and 42.3% (11/26), respectively. CONCLUSIONS: Our data on biochemical indicators demonstrates CC type gives rise to poor prognosis compared to GG type does, which suggests that close attention should be paid in the impact of adiponectin polymorphism on atherosclerotic plaque compositions.

[The role of NF-κB signaling pathway in laryngeal leukoplakia recurrent with laryngeal reflux].

Objective: To study the mechanism of vocal mucosal barrier damage mediated by NF-κB and NF-κB-regulated signaling pathway via probing the expression of inflammatory factors and essential proteins for node of NF-κB signaling pathway. Methods: The patients suffering from vocal leukoplakia accompanied with larygopharyngeal reflux(LPR) were treated with oral administration of proton pump inhibitor(PPI). Mucosal specimens of vocal cord were collected from all patients before PPI treatment. And the mucosal specimens of vocal cord were collected from the patients with suspected recurrence at 8 weeks after PPI treatment. HE staining was used to observe the histopathological changes of the mucosa. ELISA was utilized to detect the levels of inflammatory factors including tumor necrosis factor(TNF)-α, interleukin(IL)-1 and IL-6. Western blot was used to detect the expression of p-p65, p-IKK and p-IκB. Immunofluorescence method was adopted to detect the entrance of p65 to cell nucleus.Data was analyzed by SPSS 23.0 software. Results: In PPI untreated group, the expressions of IL-1ß, TNF-α and IL-6 in the specimens of 8 weeks after operation were not different significantly from those obtained during operation.But in the PPI-treated group, the expressions were down-regulated.The expression of p-p65 in the middle and high grade heterogenous hyperplasia group was higher than that of low level heterogenous hyperplasia group.The difference of p65 and p-p65 expression between 8 weeks after surgery and surgery in PPI-untreated group was statistically insignificant (P>0.05). The difference of p65 expression between PPI-treated group and PPI pre-treatment group was statistically insignificant (P>0.05). The expression of p-p65 in the PPI-treated group was lower than that of the PPI pre-treatment group (P<0.05). The expressions of IL-1ß, TNF-α and IL-6 were positively related with that of NF-κB-p65. Immun of luorescence method revealed the entrance of p65 to cell nucleus in PPI pre-treatment group, which meant that NF-κB was activated. In the PPI-treated group, few activated p65 could be observed in the cell nucleu. Conclusion: The possible mechanism of vocal mucosal barrier damage in vocal leukoplakia accompanied with LPR maybe the vocal mucosal inflammation mediated by NF-κB and NF-κB-regulated signaling pathway activated with refluxed materials.

[The efficacy of ansa cervicalis anterior root for unilateral recurrent laryngeal nerve injury].

Objective: To discuss the long-term efficacy of laryngeal reinnervation using the anterior root of the ansa cervicalis in the treatment of unilateral vocal fold paralysis (UVFP) caused by thyroid surgery. Method: From January 2010 to January 2016, a total of 39 UVFP patients who underwent ansa cervicalis anterior root-to-recurrent laryngeal nerve (RLN) anastomosis and who had suffered nerve disfunction for 6 to 24 months were enrolled as UVFP group.Another 39 age and gender matched normal subjects served as control group. Videostroboscopy, vocal function assessment (acoustic analysis, perceptual evaluation and maximum phonation time), and laryngeal electromyography were performed preoperatively and postoperatively for assessing surgery efficacy. Paired sample t test was used for statistical analysis. Result: Videostroboscopic reports indicated that the glottic closure, vocal fold edge, vocal fold position, phase symmetry and regularity were significantly improved in the UVFP group (P<0.01, respectively, postoperative vs. preoperative)and showed no statistical differences compared to the control group (P>0.05, respectively). Both the postoperative GRBAS assessment and acoustic parameters were also significantly improved in the UVFP group, Pre-operative acoustic parameters/Post-operative acoustic parameters were 1.68±0.82/0.39±0.27, 10.08±2.56/4.58±2.96, 0.203±0.216/0.018±0.038, 5.96±1.92/17.42±4.11(P<0.01, respectively) and Pre-operative acoustic parameters/Post-operative acoustic parameters were 0.39±0.27/0.32±0.19, 4.58±2.96/3.32±1.27, 0.018±0.038/0.014±0.027, 17.42±4.11/18.76±5.29, which showed no statistical differences compared to the control group (P>0.05, respectively). Conclusion: Delayed laryngeal reinnervation with the anterior root of ansa cervicalis, it can restore the physiological laryngeal phonatory function to the normal or a nearly normal voice quality, which is a feasible and effective approach for the treatment of thyroid surgery-related UVFP.

[Recent advances in early diagnosis of head and neck cancer in precision medicine era].

Head and neck squamous cell carcinoma, HNSCC, has high morbidity and mortality. Even in America, more than 1/2 to 2/3 patients have been diagnosed at advanced stage. So, it's urgent to find ways to diagnose HNSCC earlier and foresee the curative effect. With the achievement of Next-Generation Sequencing, researchers are trying to develop early diagnostic technology from a new perspective, such as personal genomics, proteomics, metabolomics and other related personal information. Here we reviewed the recent research in early diagnosis of HNSCC.

[Treatment of bilateral vocal cord paralysis by hemi-phrenic nerve transfer].

Objective: To investigate the surgical effect of reinnervation of bilateral posterior cricoarytenoid muscles(PCA) with left hemi-phrenic nerve and endoscopic laser arytenoid resection in bilateral vocal cord fold paralysis(BVFP) and to analyze the pros and cons of the two methods. Methods: One hundred and seventeen BVFP patients who underwent reinnervation of bilateral PCA using the left hemi-phrenic nerve approach (nerve group, n=52) or laser arytenoidectomy(laser group, n=65) were enrolled in this study from Jan.2009 to Dec.2015.Vocal perception evaluation, video stroboscopy, pulmonary function test and laryngeal electromyography were preformed in all patients both preoperatively and postoperative1y.Extubution rate was calculated postoperative1y. Results: Most of the vocal function parameters in nerve group were improved postoperatively compared with preoperative parameters, albeit without a significant difference(P>0.05), while laser group showed a significant deterioration in voice quality postoperative1y(P<0.05). The two groups showed significant difference in voice quality postoperative1y(P<0.05). Videostroboscopy showed that vocal fold on the operated side in both groups could abduct to various extent postoperatively, which showed significant difference when compared with preoperative abductive movements (P<0.05). But the amplitude in nerve group was larger than that in laser group (P<0.05). 89% of the patients in nerve group were inhale physiological vocal cord abductions. Postoperative glottal closure showed no significant difference in nerve group (P>0.05), while showed various increment in laser group(P<0.05). Differences between the two groups were statistically significant(P<0.05). The pulmonary function in both groups was better after operation, reaching the reference value. Postoperative laryngeal electromyography confirmed successful reinnervation of the bilateral PCA muscles. The decannulation rate were 88.5% and 81.5% in nerve group and laser group respectively. In both groups, patients presented aspiration symptoms postoperatively, and rdieved soon, except 2 patients in laser group suffered repeated aspiration. Conclusions: Reinnervation of bilateral PCA muscles using left hemi-phrenic nerve can restore inspiratory vocal fold abduction to a satisfactory extent while preserving phonatory function at the preoperative level without evident morbidity, and do not affect swallowing function, greatly improving the quality of life of the patients.

[Coblation and silicon stent implantation for recurrent laryngeal papilloma with extensive lesions].

Objective:To evaluate the feasibility and long-term outcome of radiofrequency coblation combined with silicon stent implantation in the treatment of recurrent laryngeal papilloma with extensive lesions. Method:From March 2009 to April 2016, a total of 13 patients (8 males, 5 females; aged 28-67 years) who suffered recurrent laryngeal papilloma were enrolled in this retrospective study. All the cases had undergone at least one operation at other hospitals before this hospitalization. The recurrence interval (RI) of these cases ranged from 2 to 6 months. Video-laryngoscopy or stroboscopy were preformed preoperatively and postoperatively, as well as vocal function assessment. These cases underwent radiofrequency coblationin combination of different stent implantation (member, 7 cases; tube, 4 cases; T-shaped tube, 2 cases) under general anesthesia. The follow-up was 12 to 76 months. Result:Laryngoscopy revealed that the lesions mainly located in the vocal folds, anterior commissure, ventricular band, posterior commissure, epiglottis and even in subglottic area. Varying extent of mucosal adhesion of anterior or/and posterior commissure were observed in all cases, and two cases suffered mild subglottic tracheal stenosis. Two to six weeks after surgery, the silicon stent were removed and no mucosal adhesion were found except for 2 cases who suffered mucosal adhesion of anterior commissure again. Compared to preoperative scores, VHI-10 and G scores showed the significant improvement of voice quality postoperatively in all cases. The recurrence of papilloma was observed in 3 cases during 1-year follow-up, and 4 cases during 2-year follow-up, no recurrence in 6 cases. These recurrence lesions mainly located in ventricular band, subglottic area and trachea. However, no recurrence occurred in these cases who received ablation again. No serious complications were observed in these cases. Conclusion:Radiofrequency ablation may be an effective approach in the treatment of recurrent laryngeal papilloma with extensive lesions. One-stage application of suitable silicon stents can effectively prevent the adhesion of the wound and the onset oflaryngo-tracheal stenosis, and improve the quality of voice.

Tensile Strength of Liquids: Equivalence of Temporal and Spatial Scales in Cavitation.

It is well known that strain rate and size effects are both important in material failure, but the relationships between them are poorly understood. To establish this connection, we carry out molecular dynamics (MD) simulations of cavitation in Lennard-Jones and Cu liquids over a very broad range of size and strain rate. These studies confirm that temporal and spatial scales play equivalent roles in the tensile strengths of these two liquids. Predictions based on smallest-scale MD simulations of Cu for larger temporal and spatial scales are consistent with independent simulations, and comparable to experiments on liquid metals. We analyze these results in terms of classical nucleation theory and show that the equivalence arises from the role of both size and strain rate in the nucleation of a daughter phase. Such equivalence is expected to hold for a wide range of materials and processes and to be useful as a predictive bridging tool in multiscale studies.

[Comparing the free fasciocutaneous flap with free jejunium in reconstruction for hypopharyngeal and cervical esophageal defects].

Objective:To investigate the application of the reconstruction methods for hypopharyngeal and cervical esophageal defects due to the resection of hypopharyngeal cancer and advanced laryngeal cancer between free fasciocutaneous flaps and free jejunium transfer.We compared the superiorities and inferiorities of these two reconstruction methods. Method:Retrospective review of the archives of 56 patients from 2000 to 2010 who underwent pharyngoesophageal reconstruction with free flaps (n=32) or free jejunal transfer(n=24),comparison of indications,complications, hospitalization duration, swallowing function recovery and postoperative survival time. Result:The overall 3 year survival rate of free flap group and free jejunal transfer group was 59.3%,55.7% respectively; the overall 5 year survival rate was 38.5%,37.1% respectively. The overall rate of complication rate was 18.8%, 16.7% respectively. The patients with free flaps had higher incidence rate of fistula and scarring in the donor site and lower incidence rate of hues and stricture than the ones with free jejunal transfers. The mean hospitalization duration was (15.00±7.06) days and(13.00±6.75) days. The mean time of first oral food intake was(13.00±5.83)days and (11.00±6.67) days. The differences between two groups had no statistical significance(P>0.05). Conclusion:Free flaps and free jejunium transfer are the two most common reconstruction methods for the hypopharyngeal and cervical esophageal defects. Each has its own advantages and disadvantages respectively. We should choose reconstruction method according to the site and extent of the hypopharyngeal and cervical esophageal defects, preoperative and postoperative radiotherapy requirement.

[Relationship between mutation of tumor suppressor gene p53 and development of hepatocellular carcinoma].

Fourty-five cases of primary hepatocellular carcinoma (PHC) from areas of low aflatoxin B1 exposure but high risk of hepatitis B (HBV) were examined. Positive cases of HBV-DNA were determined in 40 of 45 cases by southern hybridization and polymerase chain reaction (PCR). Results of restriction fragment length polymorphism (RFLP) analysis showed that 1 of 33 informative cases revealed loss of heterozygosity at p53 gene locus. The point mutation of p53 exon 7 was detected by PCR and restriction enzyme digestion with RsaI, MspI and HaeIII, respectively. Only two of the 45 cases showed point mutation on codon 247 and 249, and sequence analysis showed that the changes were C-->G and G-->T, respectively. Frozen section examinations in 1 of 5 cases showed weak positive staining on nucleus by immunohistochemistry with anti-p53 monoclonal antibody (pAb1801). Our results provide evidence suggesting that HBV infection alone do not contribute to changes of p53 gene including allelic loss and point mutation. A multiple step process may exist and multiple genes may be involved in hepatocarcinogenesis.