Precise Focal Spot Positioning on an Opaque Substrate Based on the Diffraction Phenomenon in Laser Microfabrication.

The precise positioning of the laser focal spot on the substrate is an important issue for laser microfabrication. In this work, a diffraction pattern-based focal spot positioning method (DFSPM) is proposed to achieve the precise positioning of the laser focal spot on opaque substrates. A series of diffraction patterns of laser focus under-positioning, exact positioning and over-positioning were obtained to investigate the cross-section light distribution of the laser focal spot. According to the monotonic tendency of FWHM to exhibit light intensity at the focal spot cross-section away from the focal plane, the FWHM threshold of polynomial fitted curves was used to determine the exact positioning of laser focus. The ascending scanning method was used to obtain the diffraction patterns at various vertical positions and the FWHM threshold of light distribution at the exact position. The polynomial fitted curves verify the FWHM monotonic tendency of light intensity distribution at the focal spot cross-section along the optical axis. Precise positioning can be achieved with a 100 nm adjustment resolution. This work was expected to provide references for laser microfabrication on opaque materials.

One stage reconstruction of mid-face fistulous defects after maxillary sinus carcinoma resection with chimeric perforator free flaps.

BACKGROUND: The reconstruction of large fistulous defects following the radical ablation of maxillary sinus carcinoma remains challenging. The procedure requires not only the coverage of both intra-nasal lining and cheek skin but also sufficient obliteration of dead space between the two surfaces. In this report, we present our experience on the reconstruction of through-and-through defects in the mid-face with poly-foliated chimeric perforator flaps. METHODS: Nine patients (five males and four females) who received a two-skin paddled and one muscle segment chimeric perforator flap reconstruction after maxillary sinus carcinoma ablation between March 2015 and December 2019 were retrospectively reviewed in authors' hospital. The mean age of the patients was 59.11. Six patients were diagnosed as squamous cell carcinoma, two as adenoid cystic carcinoma, and one as adenocarcinoma. Brown class IIIa defects were found in eight patients, and one patient had a Brown class IVa defect. The mean size of intra-nasal defect was 5.67 × 4.06 cm2 , and the mean size of facial skin defect was 8.94 × 6.56 cm2 . ALT flaps were used in five patients, LD flaps in four patients. The minor skin paddle was firstly inset to the mucosal defect site as the lining. Then, the muscle segment was inset to eliminate the dead cavity. Finally, the major skin paddle was inset to recover the cutaneous defect. RESULTS: In ALT group, the mean size of the minor skin paddle was 5.7 × 4.7 cm2 , and the mean size of the major skin paddle was 8.7 × 6.6 cm2 . In LD group, the mean size of the minor skin paddle was 6.88 × 4.38 cm2 , and the mean size of the major skin paddle was 11 × 7.75 cm2 .All donor sites were closed primarily. All flaps survived and no partial flap loss was encountered. The mean follow-up time was 14.67 months, and there were no major postoperative complications. CONCLUSION: The use of poly-foliated chimeric perforator free flaps can provide functional and aesthetic coverage for extensive through-and-through mid-face defects without significant donor-site morbidities.

Efficacy of tinnitus retraining therapy in the treatment of tinnitus: A meta-analysis and systematic review.

PURPOSE: To evaluate the efficacy of Tinnitus Retraining Therapy (TRT) in the treatment of tinnitus. MATERIALS AND METHOD: Computer retrieval of PubMed, Embase, The Cochrane Library, Web of Science, China National Knowledge Internet (CNKI), Wanfang data, etc., were conducted. According to the inclusion and exclusion criteria, the literature's quality was evaluated, and useful data was extracted. All statistical analyses were performed by RevMan5.3 software. RESULTS: 13 eligible RCTs with a total of 1345 patients were included in this meta-analysis. The meta-analysis results showed that the 1-month response rate, 3-month response rate, 6-month response rate, and overall response rate of TRT with drugs for tinnitus were higher than that of drugs only (P < 0.05). The results demonstrated that the THI scale after the treatment period of TRT with medications for tinnitus was lower than that of drugs only (P < 0.05). CONCLUSIONS: Analysis of limited studies low-quality evidence with a high risk of bias showed that the TRT was an effective treatment for tinnitus, which could improve the response rate of tinnitus and reduce the THI scale. However, more multicenter RCTs with a large sample number and high quality should verify the conclusion mentioned above.

ACP-5862 suppresses esophageal squamous cell carcinoma growth through inducing apoptosis via activation of endoplasmic reticulum stress and ROS production.

Esophageal squamous cell carcinoma (ESCC) is a common type of human oral malignancy with poor survival. Presently, it is necessary to find new and effective drugs for clinical therapy. This study aimed to identify the potential anti-tumor effects of ACP-5862, a major metabolite of acalabrutinib, on human ESCC progression, and to reveal the underlying mechanisms. Our findings suggested that ACP-5862 treatments markedly reduced the cell proliferation of ESCC cell lines in a time- and dose-dependent manner, while had no significant cytotoxicity to normal cells. Cell cycle arrest in G2/M phase was markedly induced by ACP-5862 in ESCC cells. Furthermore, apoptosis and endoplasmic reticulum (ER) stress were detected in ESCC cells treated with ACP-5862. Intriguingly, ACP-5862-induced apoptotic cell death was partly dependent on ER stress. Moreover, reactive oxygen species (ROS) was greatly triggered in ACP-5862-incubated ESCC cells, which was closely involved in apoptosis and ER stress mediated by ACP-5862. In addition, we showed that the expression of nuclear factor-erythroid 2-related factor-2 (Nrf-2) was considerably reduced in ACP-5862-treated cells. Importantly, ACP-5862 combined with Nrf-2 knockdown could further induce apoptosis and ER stress in ESCC cells compared with the ACP-5862 single group. Animal studies confirmed that repressing Nrf-2 promoted the anti-tumor effect of ACP-5862 on ESCC growth. Taken together, these findings demonstrated that ACP-5862 exerted anti-cancer effects on ESCC through inducing ER stress-mediated apoptosis via the ROS production. Meanwhile, ACP-5862 co-treated with Nrf-2 inhibitors may supply new and effective therapeutic strategies for ESCC treatment in future.

miR-218 overexpression suppresses tumorigenesis of papillary thyroid cancer via inactivation of PTEN/PI3K/AKT pathway by targeting Runx2.

BACKGROUND: It was previously reported that downregulation of miR-218 promoted thyroid cancer cell invasion, migration, and proliferation. However, the biological functions of miR-218 and its possible regulatory mechanisms in papillary thyroid cancer (PTC) cells are still elusive. MATERIALS AND METHODS: The expression levels of miR-218 and Runx2 in PTC tissues and cells were determined by quantitative real-time PCR (qRT-PCR) and Western blot. The effects of miR-218 overexpression on cell viability, invasion, apoptosis, and PTEN/PI3K/AKT pathway in PTC cells were evaluated by cell counting kit-8 assay, Transwell invasion assay, flow cytometry assay, and Western blot, respectively. Luciferase reporter assay and qRT-PCR were performed to identify the target of miR-218. Xenograft tumor experiment was performed to confirm the biological roles of miR-218 and its potential mechanisms in vivo. RESULTS: miR-218 expression was downregulated and Runx2 expression was upregulated in PTC tissues and cells. Overexpression of miR-218 suppressed viability and invasion, and induced apoptosis of PTC cells in vitro, while Runx2 overexpression greatly abolished these effects. miR-218 overexpression inactivated the PTEN/PI3K/AKT pathway, which was abated by Runx2 upregulation. Additionally, Runx2 was validated to be a direct target of miR-218. Moreover, enforced expression of miR-218 inhibited tumor growth and Runx2 expression, and blocked PTEN/PI3K/AKT pathway in vivo. CONCLUSION: miR-218 overexpression suppresses the tumorigenesis of PTC via downregulating PTEN/PI3K/AKT pathway by targeting Runx2, which indicates that miR-218 may be a potential therapeutic target for human PTC.

Molecular mechanisms underlying the protective effects of hydrogen-saturated saline on noise-induced hearing loss.

OBJECTIVES: This study aimed to explore the molecular mechanism of the protective effects of hydrogen-saturated saline on NIHL. METHODS: Guinea pigs were divided into three groups: hydrogen-saturated saline; normal saline; and control. For saline administration, the guinea pigs were given daily abdominal injections 3 d before and 1 h before noise exposure. ABR were tested to examine cochlear physiology changes. The changes of 8-hydroxy-desoxyguanosine (8-HOdG), interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α), intercellular cell adhesion molecule-1 (ICAM-1) and high mobility group box-1 protein (HMGB1) in the cochlea were also examined. RESULTS: The results showed that pre-treatment with hydrogen-saturated saline could significantly attenuate noise-induced hearing loss. The concentration of 8-HOdG was also significantly decreased in the hydrogen-saturated saline group compared with the normal saline group. After noise exposure, the concentrations of IL-1, IL-6, TNF-α, and ICAM-1 in the cochlea of guinea pigs in the hydrogen-saturated saline group were dramatically reduced compared to those in the normal saline group. The concentrations of HMGB-1 and IL-10 in the hydrogen-saturated saline group were significantly higher than in those in the normal saline group immediately and at 7 d after noise exposure. CONCLUSIONS: This study revealed for the first time the protective effects of hydrogen-saturated saline on noise-induced hearing loss (NIHL) are related to both the anti-oxidative activity and anti-inflammatory activity.

A POU3F4 Mutation Causes Nonsyndromic Hearing Loss in a Chinese X-linked Recessive Family.

BACKGROUND: The molecular genetic research showed the association between X-linked hearing loss and mutations in POU3F4. This research aimed to identify a POU3F4 mutation in a nonsyndromic X-linked recessive hearing loss family. METHODS: A series of clinical evaluations including medical history, otologic examinations, family history, audiologic testing, and a high-resolution computed tomography scan were performed for each patient. Bidirectional sequencing was carried out for all polymerase chain reaction products of the samples. Moreover, 834 controls with normal hearing were also tested. RESULTS: The pedigree showed X-linkage recessive inheritance pattern, and pathogenic mutation (c.499C>T) was identified in the proband and his family member, which led to a premature termination prior to the entire POU domains. This mutation co-segregated with hearing loss in this family. No mutation of POU3F4 gene was found in 834 controls. CONCLUSIONS: A nonsense mutation is identified in a family displaying the pedigree consistent with X-linked recessive pattern in POU3F4 gene. In addition, we may provide molecular diagnosis and genetic counseling for this family.

[Clinical application of heterogeneous acellular dermal matrix with autologous bone meal in open tympanoplasty].

OBJECTIVE: To evaluate the clinical value of heterogeneous acellular dermalmatrix with autologous bone meal in open tympanoplasty. METHOD: Twenty-eight cases (30 ears) with middle ear cholesteatoma were trea- ted by open tympanoplasty and repaired by heterogeneous acellular dermalmatrix autologous bone meal on study team. Twenty-two cases (22 ears) with middle ear cholesteatoma were treated by open tympanoplasty on control team. All patients were followed up for 12 to 18 months and assessed the fuction postoperatively. RESULT: The re- construction of external auditory canal structure is close to normal, and no narrow happens on study team. The rate of dry ear was about 90%. All cases had no recurrence of cholesteatoma. CONCLUSION: Application of decellu- larized dermal matrix with autologous bone meal can rise early to cover the wound, promote wound healing and to reduce the external auditory canal, reduce the effect of granulation and scar formation. It is a kind of method of repair to be promoted.

Phenotype-genotype correlation in 295 Chinese deaf subjects with biallelic causative mutations in the GJB2 gene.

AIMS: The connexin 26 coding gene (GJB2) is the primary causative gene for nonsyndromic sensorineural hearing impairment (NSSHI). More than 100 mutations in this gene have been reported to be linked to hearing impairment (HI), from mild to profound hearing loss. To precisely estimate the impact of GJB2 mutations in the Chinese population, a cross-sectional study was performed to analyze the auditory data of Chinese patients with NSSHI. RESULTS: Two hundred ninety-five unrelated patients with NSSHI with biallelic mutations in GJB2 were recruited from seven provinces in Northern China from 2004 to 2008. The levels of HI and average pure tone audiometry were compared across different genotypes by χ(2) testing. The subjects with the genotypes of combined truncating mutations had more cases of severe HI than the subjects with a genotype of several nontruncating mutations. It was also revealed that subjects carrying either c.[79G>A; 341A>G]+[79G>A; 341A>G] or c.[109G>A]+[79G>A; 341A>G] had significantly fewer cases of severe HI than the reference group of homozygous c.235delC, whereas the subjects carrying c.[235delC]+[176_191del16] had more cases of severe HI than the homozygous c.235delC group. CONCLUSIONS: This is the first study to clarify the correlations between different GJB2 biallelic genotypes and NSSHI phenotype in the Chinese population. The Chinese subjects with two truncating mutations in GJB2 were shown to correlate with more severe HI.

The large Chinese family with Y-linked hearing loss revisited: clinical investigation.

CONCLUSION: The DFNY1 phenotypes shared many characteristics with some autosomal dominant hearing loss, in the aspects of age of onset, severity and audiometric configuration. However, the typical, outstanding feature of this trait was its remarkable pattern of inheritance. Similar traits, if ever encountered, can be most easily identified by discerning this exceptional and rare pattern of inheritance. OBJECTIVES: To analyze the audiological features in Chinese Y-linked non-syndromic hearing impairment, the extended DFNY1 family. SUBJECTS AND METHODS: A nine-generation Chinese family (DFNY1) was ascertained and expanded from the year of 2000 to 2006. The audiometric evaluations included pure-tone audiometry, tympanometry, and auditory brainstem responses. Some subjects received computerized tomography scan of the temporal bone. RESULTS: 52 out of 276 members in this family received clinical examinations. 24 live subjects had hearing impairment consisting of 23 patrilineal males and one female. In the affected lineage, 92% patrilineal males were well characterized as having hearing loss and 2 children remained to be diagnosed. Based on the audiological examinations on the male members, the degree of hearing loss was from mild (3 patients), moderate (7 patients) to severe (11 patients). The audiometry displayed 48% subjects with sloping in high frequencies, 38% flat in all frequencies, and the rest (14%) the U-shape. The age of onset ranged from 5-27 years with the average of 11.5 years.

[Correlation between phonetically balanced maximum and pure tone auditory threshold among 106 auditory neuropathy patients].

OBJECTIVE: To estimate correlation between phonetically balanced maximum (PB max) and pure tone auditory threshold in auditory neuropathy (AN) patients. METHODS: One hundred and six AN patients were identified using multiple criteria including PB max, a metric for speech recognition, pure tone auditory threshold, acoustic emission test, distortion products otoacoustic emission (DPOAE) and auditory brainstem response (ABR). SPSS statistical software was used to estimate the Pearson's correlation between PB max and pure tone auditory threshold and to test whether pure tone auditory threshold, or auditory configuration had a significant impact on PB max. RESULTS: Even the patients had the same or similar values for pure tone auditory threshold or auditory configuration, varied values of PB max were found in two hundreds and twelve ears for 106 patients. Analysis of the data for 106 patients revealed a negative correlation (r = -0. 602, P <0. 01) between PB max and pure tone auditory threshold, i. e. hearing loss at a mild relates to a lower PB max. By using analysis of variance (ANOVA) method, it was found that both pure tone auditory threshold and auditory configuration had a significant (P <0.01) impact on the patients' PB max. CONCLUSIONS: This analysis implicated the promise and potential of pure tone auditory threshold and auditory configuration for predicting PB max of the AN patients, and improving the diagnosis of AN.

GJB2, SLC26A4 and mitochondrial DNA A1555G mutations in prelingual deafness in Northern Chinese subjects.

CONCLUSION: This genetic epidemiological study demonstrated that 26.65% of the prelingual deafness in Northern Chinese patients can be detected at younger ages by genetic testing of three common hearing loss genes (GJB2, SLC26A4 and mtDNA A1555G), and thus, early intervention measures could be undertaken to help them in language acquisition. OBJECTIVES: The GJB2, SLC26A4 and mtDNA A1555G mutations are the prevalent causes of prelingual deafness worldwide. Numerous studies have revealed that the forms and frequencies of the mutations in the three genes are largely dependent on the ethnic or geographic origins. Hence, this study aimed to characterize the mutation profiles of the three genes in prelingual deafness in Northern Chinese patients. SUBECTS AND METHODS: An investigation of 514 patients with prelingual deafness and 117 controls with normal hearing was conducted. Bidirectional sequencing (or enzyme digestion) was applied to identify sequence variations. RESULTS: This study revealed that 26.65% patients had two mutated alleles (homozygote or compound heterozygote) of GJB2 (9.14%) or SLC26A4 (8.95%) and/or an mtDNA A1555G (8.56%) mutation. In detail, 19.26% patients carried GJB2 mutations including 10.12% single mutant carriers. 235delC was the most common type, making up 69.18% of all mutants for GJB2. The mutant carrier rate for SLC26A4 was 15.2%, including 6.23% single mutant carriers. The two most common types (IVS7-2A > G and H723R) accounted for 51.61% and 33.06% mutations, respectively. Forty-five patients had mtDNA A1555G, giving a frequency of 8.75%. In the control group with normal hearing, 2.56%, 1.71% and 0% of the subjects carried a single mutant for GJB2, SLC26A4 and mtDNA A1555G, respectively.

[Studies of the strategy for newborn gene screening].

OBJECTIVE: To discuss and analyze the feasibility and strategy for perform the newborn gene screening in the process of newborn hearing screening in order to supply the defects or limitation in the hearing screening. METHODS: Four hundreds and sixty newborn babies from December 2006 to April 2007 accepted the simultaneous hearing and gene screening. Otoacoustic emission (OAE) was used for the first step hearing screening and OAE combined with auto auditory brainstem response (AABR) detection for the second step screening. Newborn genetic disease screening cards were used for collecting the blood spot from the umbilical cord within the moment of newborn. The cards could be directly performed the polymerase chain reaction (PCR) for screening the mitochondrial 12SrRNA 1555G and GJB2 as well as SLC26A4 genes mutations. The restriction enzyme Alw26I was used to recognize the point mutation of 12SrRNA A1555G. The samples with the possible 12SrRNA A1555G mutation were then sequenced to verify. The PCR products from the GJB2 coding region and SLC26A4 IVS7-2A > G hot spot region were sequenced directly. The software of DNAStar was used to analysis the sequence. RESULTS: The first step of hearing screening of 460 newborn babies showed " refer" on the left ear of nine babies and on the right ear of three babies. Seven showed "refer" on bilateral with the the total of babies 19. After 42 days, they accepted the second step for hearing screening. 16 of the 19 were showed "pass" with OAE and AABR. One baby showed "pass" on the left ear, "refer" on the right ear with the OAE detection but bilateral "pass" with AABR. Two babies failed to accept the re-examination. The newborn gene screening showed five of the 460 babies had the positive response on the A1555G restriction enzyme assay. Of the five babies, one was proved to be the 12SrRNA A1555G mutation and three were the C1556T mutations and one sequence was normal. For the SLC26A4 gene screening, five were the heterozygote of IVS7-2A > G mutation were found and one was carrier the polymorphism of IVS7-18T > G and another was IVS6-62_63insGT heterozygote carrier. For the GJB2 gene screening, eight were 235delC heterozygote carriers, four were G109A heterozygote carriers. All the gene screening found 23 newborn babies of the 460 harbored the changes in the three genes. Of those, one was the 12SrRNA A1555G. pathogenic mutation and 13 were pathogenic heterozygote carriers, nine were the polymorphisms. It was worth to pay more attentions that A1555G mutation was found in the baby whose hearing screening was "pass" in the hearing screening as well as the 13 heterozygote carrier for GJB2 and SLC26A4 gene. CONCLUSIONS: It might be one of the powerful strategy for adding the concept of newborn gene screening into the hearing screening for the purpose of early diagnosis and discovery the prelingual or late-onset or the high risk as well as the pathogenic carriers. On the basis of the research progress, it was necessary to develop the national newborn gene screening into the process of newborn hearing screening.

[Construction of an expression vector under the control of hTERT promoter].

AIM: To construct a luciferase expression vector driven by hTERT(human telomerase reverse transcriptase) core promoter and identify the transcriptional activity of the vector in tumor cells and normal cells. METHODS: hTERT gene core promoter was amplified by PCR using the total genomic DNA from HeLa cells as template. The amplified gene fragment was subsequently cloned into PGL3-basic vector. Then the expression vector was confirmed by restriction enzyme digestion and PCR analysis. The luciferase activity driven by the hTERT gene core promoter was identified after transient transfection of the expression vector into tumor cells and normal cells. RESULTS: The luciferase activity was high in the transfected tumor cells, and very low in transfected normal cells. CONCLUSION: hTERT gene core promoter is tumor-specific and may be useful in gene therapy of tumor.

[Apoptosis of laryngeal carcinoma cell line Hep2 induced by tumor-specific expression of TRAIL gene].

AIM: To explore targeted gene therapy of tumor by using the combination of TRAIL gene with the telomerase promoter. METHODS: TRAIL gene with an IL-2 signal peptide was constructed by PCR and cloned into vector pGL3-181hTERT downstream of hTERT promoter to form an eukaryotic expressing vector. Hep2 cells were transfected by the recombinant vector and apoptosis of the transfected cells was evaluated by trypan-blue exclusion and the agarosegel electrophoresis of DNA. RESULTS: We successfully constructed a recombinant eukaryotic expression vector for TRAIL gene.The expressed product significantly induced the apoptosis of Hep2 cells. CONCLUSION: The recombinant eukaryotic expression vector pGL3-181hTERT/TRAIL was successfully constructed, which provides the possibility for gene therapy of tumor.