Significant improvements in the olfactory sensitivity of bipolar I disorder patients during euthymia versus manic episodes: a longitudinal study.

Introduction: Research has indicated that individuals diagnosed with bipolar disorder (BD) might experience alterations in their olfaction or levels of serum tumor necrosis factor-α (TNF-α), but no studies have investigated olfactory function and serum TNF-α in BD patients simultaneously. Moreover, there is a lack of existing research that compares the longitudinal olfactory function between individuals with manic and euthymic BD I. Methods: Patients with manic BD I (BDM, n=44) and healthy controls (HCs, n=32) were evaluated symptoms (measured via the Young Manic Rating Scale, YRMS), social function (measured via the Global Assessment Function, GAF), serum TNF-α, and olfactory function (via the Sniffin' Sticks test) including olfactory sensitivity (OS) and olfactory identification (OI). The BDM patients were followed up to the remission period and re-evaluated again. We compared OS, OI and serum TNF-α in manic and euthymic patients with BD I and HCs. We examined the correlation between olfactory function and symptoms, social function, and serum TNF-α in patients with BD I. Results: The BDM patients exhibited significantly lower OS and OI compared to the HCs (Z = -2.235, P = 0.025; t = -6.005, P < 0.001), while a positive correlation was observed between OS and GAF score (r = 0.313, P = 0.039). The OS in the BD I remission group (n=25) exhibited significantly superior performance compared to the BDM group (t = -4.056, P < 0.001), and the same as that in the HCs (P = 0.503). The change in OS showed a positive correlation with the decrease in YMRS score (r = 0.445, P = 0.026), and a negative correlation with the course of disease (r = -0.594, P = 0.002). The TNF-α in BD I patients was significantly lower compared to HCs (P < 0.001), and not significantly correlated with olfactory function (all P > 0.05). Conclusion: The findings suggest that OS and OI are impaired in BDM patients, and the impaired OS in those patients can be recovered in the remission stage. OI may serve as a potential characteristic marker of BD. OS might be useful as an index for BDM treatment efficacy and prognosis.

Comparison of olfactory function, cognitive function and serum tumor necrosis factor-α between bipolar and schizophrenic patients in the remission stage.

OBJECTIVES: Olfactory function, serum tumor necrosis factor-α (TNF-α) and cognitive function were compared between bipolar disorder (BD) and schizophrenia (SP) patients in the remission stage combined with correlation analysis, with the aim of identifying new indicators for the auxiliary diagnosis of these psychiatric illnesses. METHODS: A total of 46 euthymic BD patients, 42 clinically stable SP patients and 42 healthy controls (HC) were included in this study. Olfactory sensitivity (OS) and olfactory identification (OI) were assessed using Sniffin' Sticks test, and serum TNF-α levels were measured by ELISA. Clinical symptoms were evaluated with the Hamilton Rating Scale for Depression, Young Mania Rating Scale, Hamilton anxiety scale, and the Positive and Negative Syndrome Scale (PANSS). Social function was evaluated with the Global Assessment Function (GAF) scale. Cognitive function was evaluated using the Trail Making Test-A (TMT-A) and Digit Cancellation Test (DCT). RESULTS: OI and cognitive function scores and serum TNF-α levels were significantly lower in the BD and SP patients compared with the HC participants. There was no significant difference between the BD and SP groups, and there were no significant differences in OS among the three groups. OI score was positively correlated with years of education in both the BD and SP groups. OI score in the SP group was negatively correlated with age and PANSS score, and positively correlated with GAF score. In the BD group, OS was positively correlated with DCT II and DCT III. In the SP group, OS and OI scores were positively correlated with DCT III, and negatively correlated with TMT-A time. Furthermore, there was a positive correlation between TNF-α and DCT II in the BD group. There was no significant linear correlation between olfactory function and TNF-α in the BD or SP group. CONCLUSION: OI may be a trait marker for BD and SP. Some cognitive functions may be correlated not only with TNF-α in BD patients in remission, but also with olfactory function in BD and SP patients in remission.

Impaired olfactory function in bipolar disorder patients during acute episodes regardless of psychotic symptoms.

Background: The aim of this study was to analyze whether the presence of psychotic symptoms affects olfactory function in patients with bipolar disorder (BD). We also compared olfactory function between the period of episode and remission in patients with BD. Methods: BD patients in the acute phase were tracked to the remission stage. The psychiatric symptoms and social function of the enrolled subjects were assessed using the Hamilton Rating Scale for Depression (HAMD), the Young Mania Rating Scale (YMRS), the Hamilton Rating Scale for Anxiety (HAMA), the Positive and Negative Syndrome Scale (PANSS) and the Global Assessment Function (GAF). Olfactory sensitivity (OS) and olfactory identification (OI) was assessed using the Sniffin' Sticks test. Differences in OS and OI among the episodic group, the euthymic group, and the healthy control (HC) group were compared. According to whether BD is accompanied by psychotic symptoms, the OS and OI in the BD with psychotic symptoms group (P-BD), the BD without psychotic symptoms group (NP-BD), and the HC group were compared. Results: The P-BD and NP-BD groups exhibited impaired OI compared with the HC group, but there was no significant difference in OI between the P-BD and NP-BD groups, or in OS among all three groups. All patients with episodic BD had significantly lower OS and OI compared with the HC group. OI in euthymic BD patients was still impaired; however, OS recovered, showing no significant difference compared with that in the HC group. Conclusion: The results indicate that patients with episodic BD have impaired OS and OI, regardless of psychotic symptoms. OI may be a characteristic marker of BD, and OS may be a state marker that can be used to distinguish between episodic and euthymic BD.

A Novel EYA1 Mutation Causing Alternative RNA Splicing in a Chinese Family With Branchio-Oto Syndrome: Implications for Molecular Diagnosis and Clinical Application.

OBJECTIVES: Branchio-oto syndrome (BOS) primarily manifests as hearing loss, preauricular pits, and branchial defects. EYA1 is the most common pathogenic gene, and splicing mutations account for a substantial proportion of cases. However, few studies have addressed the structural changes in the protein caused by splicing mutations and potential pathogenic factors, and several studies have shown that middle-ear surgery has limited effectiveness in improving hearing in these patients. BOS has also been relatively infrequently reported in the Chinese population. This study explored the genetic etiology in the family of a proband with BOS and provided clinical treatment to improve the patient's hearing. METHODS: We collected detailed clinical features and peripheral blood samples from the patients and unaffected individuals within the family. Pathogenic mutations were identified by whole-exome sequencing and cosegregation analysis and classified according to the American College of Medical Genetics and Genomics guidelines. Alternative splicing was verified through a minigene assay. The predicted three-dimensional protein structure and biochemical experiments were used to investigate the pathogenicity of the mutation. The proband underwent middle-ear surgery and was followed up at 1 month and 6 months postoperatively to monitor auditory improvement. RESULTS: A novel heterozygous EYA1 splicing variant (c.1050+4 A>C) was identified and classified as pathogenic (PVS1(RNA), PM2, PP1). Skipping of exon 11 of the EYA1 pre-mRNA was confirmed using a minigene assay. This mutation may impair EYA1-SIX1 interactions, as shown by an immunoprecipitation assay. The EYA1-Mut protein exhibited cellular mislocalization and decreased protein expression in cytological experiments. Middle-ear surgery significantly improved hearing loss caused by bone-conduction abnormalities in the proband. CONCLUSION: We reported a novel splicing variant of EYA1 in a Chinese family with BOS and revealed the potential molecular pathogenic mechanism. The significant hearing improvement observed in the proband after middle-ear surgery provides a reference for auditory rehabilitation in similar patients.

Gene regulation analysis of patient-derived iPSCs and its CRISPR-corrected control provides a new tool for studying perturbations of ELMOD3 c.512A>G mutation during the development of inherited hearing loss.

The ELMOD3 gene is implicated in causing autosomal recessive/dominant non-syndromic hearing loss in humans. However, the etiology has yet to be completely elucidated. In this study, we generated a patient-derived iPSC line carrying ELMOD3 c.512A>G mutation. In addition, the patient-derived iPSC line was corrected by CRISPR/Cas9 genome editing system. Then we applied RNA sequencing profiling to compare the patient-derived iPSC line with different controls, respectively (the healthy sibling-derived iPSCs and the CRISPR/Cas9 corrected iPSCs). Functional enrichment and PPI network analysis revealed that differentially expressed genes (DEGs) were enriched in the gene ontology, such as sensory epithelial development, intermediate filament cytoskeleton organization, and the regulation of ion transmembrane transport. Our current work provided a new tool for studying how disruption of ELMOD3 mechanistically drives hearing loss.

A non-coding variant in 5' untranslated region drove up-regulation of pseudo-kinase EPHA10 and caused non-syndromic hearing loss in humans.

Hereditary hearing loss has a genetic and phenotypic heterogeneity. However, it is still difficult to explain this heterogeneity perfectly with known deafness genes. Here, we report a novel causative gene EPHA10 as well as its non-coding variant in 5' untranslated region identified in a family with post-lingual autosomal dominant non-syndromic hearing loss from southern China. One affected member of this family had an ideal hearing restoration after cochlear implantation. We speculated that there were probable deafness-causing abnormalities in the cochlea according to clinical imaging and auditory evaluations. A heterozygous variant c.-81_-73delinsAGC was found co-segregating with hearing loss. Epha10 was expressed in mouse cochlea at both transcription and translation levels. The variant caused upregulation of EPHA10 which may result from promoter activity enhancement after sequence change. Overexpression of Eph (the homolog of human EPHA10) exerted effects on the structure and function of chordotonal organ in fly model. In summary, our study linked pseudo-kinase EPHA10 to hearing loss in humans for the first time.

Establishment of two iPSC lines from healthy donor with heterozygous mutation in the SLC26A4 gene.

The human induced pluripotent stem cell (iPSC) lines, CSUXHEi001-A and CSUXHEi002-A, were generated from peripheral blood mononuclear cells (PBMCs). The donors were couple and each of them has a heterozygous mutation in the SLC26A4 gene. It manifests in their children as Enlarged vestibular aqueduct (EVA). The use of iPSC will allow describing the early stages of hearing loss, which is undoubtedly relevant for identifying key stages of development at which phenotypic manifestations of mutations in the SLC26A4 gene are found.

A nonsense TMEM43 variant leads to disruption of connexin-linked function and autosomal dominant auditory neuropathy spectrum disorder.

Genes that are primarily expressed in cochlear glia-like supporting cells (GLSs) have not been clearly associated with progressive deafness. Herein, we present a deafness locus mapped to chromosome 3p25.1 and an auditory neuropathy spectrum disorder (ANSD) gene, TMEM43, mainly expressed in GLSs. We identify p.(Arg372Ter) of TMEM43 by linkage analysis and exome sequencing in two large Asian families segregating ANSD, which is characterized by inability to discriminate speech despite preserved sensitivity to sound. The knock-in mouse with the p.(Arg372Ter) variant recapitulates a progressive hearing loss with histological abnormalities in GLSs. Mechanistically, TMEM43 interacts with the Connexin26 and Connexin30 gap junction channels, disrupting the passive conductance current in GLSs in a dominant-negative fashion when the p.(Arg372Ter) variant is introduced. Based on these mechanistic insights, cochlear implant was performed on three subjects, and speech discrimination was successfully restored. Our study highlights a pathological role of cochlear GLSs by identifying a deafness gene and its causal relationship with ANSD.

The Spatiotemporal Evolution Analysis of Ecosystem Pattern in Wenchuan (Magnitude 8.0) Earthquake Disaster Area, China.

The ecological system is the basis of human survival and global environmental protection. In the process of development, countries will pay close attention to the changing state of the ecosystem. Taking the ecosystem pattern as the research object, a three-layer analysis method was proposed. The transfer matrix and landscape index were used as the first layer to analyze the basic changes. Grey correlation, range-coupling coordination and relative priority were used as the second layer to analyze the reasons of the change. The interval-entropy weight, TOPSIS (Technique for Order Preference by Similarity to an Ideal Solution), was used as the third layer to evaluate the quality of the change. The ten counties in the worst-hit areas of the Wenchuan earthquake were analyzed from different angles, with county region, intensity zone and ecosystem as the objects, and the following results were obtained: (1) Taking Mianzhu City as an example, from 2000 to 2010 and 2018, the conversion ratio of forest, grassland and farmland is 54.24, 59.19, 17.21, 20.06, 37.39 and 52.86%, which were distributed in the north, central and southern parts, respectively. (2) Taking the ninth intensity zone as an example, the forest landscape fragmentation increased, disturbance decreased, and species diversity increased. There is a high influence and restriction relationship between ecosystem and landscape pattern in the total landscape area change. Additionally, the relationship between them tends to develop in a benign way. As of 2018, it is in the change state of moderate imbalance-ecosystem lag. (3) Taking the county ecosystem change as an example, urban type is the best in the counties of ecosystem change, of which Shifang is the best and Pingwu is the worst. The results show that this method can effectively compare and analyze the changes in the multi-regional ecosystem pattern, which has the characteristics of universality and can also be applied to the research of ecosystem pattern change in special regions.

Establishment of an iPSC line (CSUXHi004-A) from a patient with Waardenburg syndrome type I caused by a PAX3 splice mutation.

Waardenburg Syndrome (WS) is a common autosomal dominant syndrome associated with hearing loss. Its clinical manifestations include hearing impairment and pigmentation anomalies. In this study, we generated an induced pluripotent stem cell (iPSC) line from the Epstein-Barr virus-immortalized B lymphocytes of a 6-year-old boy affected with WS type I, caused by a heterozygous splice site mutation in the PAIRED BOX GENE 3 (PAX3) (NM_181457.3: c.452-2A > G). The patient-specific iPSC line (CSUXHi004-A) carrying the same PAX3 mutation showed a normal karyotype, expressed pluripotent markers, and presented differentiation capacity in vitro. This method may be a useful tool for the in vitro modeling of WS.

Coupled-mode induced transparency via Ohmic heating in a single polydimethylsiloxane-coated microbubble resonator.

We demonstrate an approach for the realization of coupled-mode induced transparency (CMIT) in a hybrid polydimethylsiloxane (PDMS)-coated silica microbubble resonator, with an Au microwire inserted in the hollow channel. Owing to the large negative thermo-optics coefficient of PDMS, different radial order modes with opposite thermal sensitivities can coexist in this hybrid microcavity. By applying a current through the Au microwire, which acts as a microheater, the generated Ohmic heating could thermally tune the resonance frequencies and the frequency detuning of the coupled mode to achieve controllable CMIT. This platform offers an efficient and convenient way to obtain controllable CMIT for applications, such as label-free biosensing and quantum information processing.

Tunable optofluidic liquid metal core microbubble resonator.

This study introduces design and coupling techniques, which bridge an opaque liquid metal, optical WGM mode, and mechanical mode into an opto-mechano-fluidic microbubble resonator (MBR) consisting of a dielectric silica shell and liquid metal core. Benefiting from the conductivity of the liquid metal, Ohmic heating was carried out for the MBR by applying current to the liquid metal to change the temperature of the MBR by more than 300 °C. The optical mode was thermally tuned (>3 nm) over a full free spectral range because the Ohmic heating changed the refractive index of the silica and dimeter of the MBR. The mechanical mode was thermally tuned with a relative tuning range of 9% because the Ohmic heating changed the velocity and density of the liquid metal.

Combining whispering gallery mode optofluidic microbubble resonator sensor with GR-5 DNAzyme for ultra-sensitive lead ion detection.

Lead ions are deleterious pollutants that often reach drinking water, and can cause significant harm to humans (particularly children). An ultra-sensitive lead ion detection method using a whispering gallery mode (WGM) optofluidic microbubble resonator and the classic GR-5 DNAzyme is proposed in this paper. With the auxiliary piranha and Ploy-l-lysine solution, GR-5 DNAzyme was successfully modified on the inner wall of a microbubble. The mode field distribution of the microbubble was analysed, and the optofluidic sensor with thin wall exhibited a maximum bulk refractive index sensitivity of 265.2 nm/RIU. Lead ions at concentrations ranging from 0.1 pM to 100 pM were tested using the proposed WGM optofluidic sensor. The noise was decreased to 2.43 fM using the self-referenced differential method. Thus, a limit of approximately 15 fM was obtained for the detection of lead ions using the WGM optofluidic biosensor. Eight competing metal ions were also used to evaluate the selectivity of the proposed sensor, with results indicating that it has high selectivity for lead ions. Finally, the sensor performance is verified using real samples.

Automatic Extraction of Structural and Non-Structural Road Edges from Mobile Laser Scanning Data.

Accurate road information is important for applications involving road maintenance, intelligent transportation, and road network updates. Mobile laser scanning (MLS) can effectively extract road information. However, accurately extracting road edges based on large-scale data for complex road conditions, including both structural and non-structural road types, remains difficult. In this study, a robust method to automatically extract structural and non-structural road edges based on a topological network of laser points between adjacent scan lines and auxiliary surfaces is proposed. The extraction of road and curb points was achieved mainly from the roughness of the extracted surface, without considering traditional thresholds (e.g., height jump, slope, and density). Five large-scale road datasets, containing different types of road curbs and complex road scenes, were used to evaluate the practicality, stability, and validity of the proposed method via qualitative and quantitative analyses. Measured values of the correctness, completeness, and quality of extracted road edges were over 95.5%, 91.7%, and 90.9%, respectively. These results confirm that the proposed method can extract road edges from large-scale MLS datasets without the need for auxiliary information on intensity, image, or geographic data. The proposed method is effective regardless of whether the road width is fixed, the road is regular, and the existence of pedestrians and vehicles. Most importantly, the proposed method provides a valuable solution for road edge extraction that is useful for road authorities when developing intelligent transportation systems, such as those required by self-driving vehicles.

Elmod3 knockout leads to progressive hearing loss and abnormalities in cochlear hair cell stereocilia.

ELMOD3, an ARL2 GTPase-activating protein, is implicated in causing hearing impairment in humans. However, the specific role of ELMOD3 in auditory function is still far from being elucidated. In the present study, we used the CRISPR/Cas9 technology to establish an Elmod3 knockout mice line in the C57BL/6 background (hereinafter referred to as Elmod3-/- mice) and investigated the role of Elmod3 in the cochlea and auditory function. Elmod3-/- mice started to exhibit hearing loss from 2 months of age, and the deafness progressed with aging, while the vestibular function of Elmod3-/- mice was normal. We also observed that Elmod3-/- mice showed thinning and receding hair cells in the organ of Corti and much lower expression of F-actin cytoskeleton in the cochlea compared with wild-type mice. The deafness associated with the mutation may be caused by cochlear hair cells dysfunction, which manifests with shortening and fusion of inner hair cells stereocilia and progressive degeneration of outer hair cells stereocilia. Our finding associates Elmod3 deficiencies with stereocilia dysmorphologies and reveals that they might play roles in the actin cytoskeleton dynamics in cochlear hair cells, and thus relate to hearing impairment.

Proband Whole-Exome Sequencing Identified Genes Responsible for Autosomal Recessive Non-Syndromic Hearing Loss in 33 Chinese Nuclear Families.

Autosomal recessive non-syndromic hearing loss (ARNSHL) is a highly heterogeneous disease involving more than 70 pathogenic genes. However, most ARNSHL families have small-sized pedigrees with limited genetic information, rendering challenges for the molecular diagnosis of these patients. Therefore, we attempted to establish a strategy for identifying deleterious variants associated with ARNSHL by applying proband whole-exome sequencing (proband-WES). Aside from desiring to improve molecular diagnostic rates, we also aimed to search for novel deafness genes shared by patients with similar phenotype, making up for the deficiency of small ARNSHL families. In this study, 48.5% (16/33) families were detected the pathogenic variants in eight known deafness genes, including 10 novel variants identified in TMPRSS3 (MIM 605551), MYO15A (MIM 602666), TMC1 (MIM 606706), ADGRV1 (MIM 602851), and PTPRQ (MIM 603317). Apart from six novel variants with a truncating effect (nonsense, deletion, insertion, and splice-site), four novel missense variants were not found in 200 unrelated control population by using Sanger sequencing. It is important to note that none of novel genes were shared across different pedigrees, indicating that a larger sample size might be needed. Proband-WES is a cost-effective and precise way of identifying causative variants in nuclear families with ARNSHL. This economical strategy may be appropriated as a clinical application to provide molecular diagnostics, genetic counseling, and individualized health maintenance measures for patients with ARNSHL at hearing clinics.

[Neuroprotective effects of combined pretreatment with edaravone and propofol on neonatal rat cerebral cortical neurons with ischemia/reperfusion injury in vitro].

OBJECTIVE: To investigate the protective effect of combined pretreatment of edaravone and propofol on cerebral cortex with ischemia/reperfusion (I/R) injury and its therapeutic window. METHODS: Sprague Dawley (SD) rat brain cortex cells harvested within 24 hours of birth were cultured in vitro for 7 days. The cells were then divided into blank control group, glutamate injury group, 24-hour drug precondition control group, and 24-, 2-, 0-hour drug precondition groups according to random number table. The nerve cells in each pretreatment group were cultured in medium containing 100 µmol/L of edaravone and 3 mg/L of propofol 24, 2, or 0 hour before glutamate damage (200 µmol/L for 0.5 hour). Nerve cell survival or damage was determined by methyl thiazolyl tetrazolium (MTT), lactate dehydrogenase (LDH) leakage rate, and nerve cell Na+-K+-ATPase activity. The oxidation and anti-oxidation ability of nerve cells was observed by determining superoxide dismutase (SOD) activity (xanthine oxidase), malondialdehyde (MDA) content (thiobarbituric acid). Nerve apoptosis was detected by flow cytometry. RESULTS: Compared with blank control group, in the glutamate injury group, nerve cell survival rate [(62.2±23.4)% vs. (90.5±14.8)%], the activity of SOD (U/ml: 6.864±2.872 vs. 29.569±3.684), Na+-K+-ATPase activity [U×mg(-1)×h(-1): 0.318±0.146 vs. 0.636±0.168] were significantly decreased, and rate of neuronal apoptosis [(9.4±0.7)% vs. (6.1±0.2)%], the content of MDA (nmol/ml: 0.515±0.101 vs. 0.294±0.105), LDH leakage rate [(41.2±1.6)% vs. (36.8±4.6)%] were significantly increased (P<0.05 or P<0.01). Compared with glutamate injury group, the cell survival rate and the activity of SOD and Na+-K+-ATPase were significantly increased in the drug pretreatment groups, and apoptosis rate, MDA content, and LDH leakage rate were significantly decreased with time-department, and effect in the 24-hour pretreatment group was most significant [survival rate of cell: (89.2±30.3)% vs. (62.2±23.4)%, SOD activity (U/ml): 17.780±4.514 vs. 6.864±2.872, Na+-K+-ATPase activity [U×mg(-1)×h(-1)]: 0.541±0.052 vs. 0.318±0.146, the rate of cell apoptosis: (6.7±0.4)% vs. (9.4±0.7)%, the content of MDA (nmol/ml): 0.319±0.101 vs. 0.515±0.101, LDH leakage rate: (37.2±1.4)% vs. (41.2±1.6)%, all P<0.01]. CONCLUSION: The synergistic protective effect of pretreatment with edaravone combined with propofol on neonatal rat brain cortex cells with I/R injury in vitro was evident; and 24-hour pretreatment is the best time window of protection for the cerebral neurons.

[Scanning electron microscope observation of plerocercoid from Rana nigromaculata in Guizhou Province].

Five integral and active plerocercoids collected from naturally infected frogs were fixed by glutaraldehyde-osmic acid, dehydrated in graded series of ethanol, dried via vacuum freeze and coated with carbon gold. They were then observed by scanning electron microscopy. The results showed that the anterior and posterior ends of the plerocercoids were slightly swell and without segmentation. Annular furrows ran irregularly along the body surface. The anterior end was drawn inside and had a deep dorsoventral hollow, around which, the plasma membrane displayed a lip bulge. The posterior end was similar to the anterior one in the shape, but a fissure in its central part was comparatively narrower and shallower than the hollow in the anterior end. There were thousands of pits and grooves on the body surface, unequal in size. The whole body surface of the plerocercoid was densely covered by sharp spine-like microtriches.

Investigation of cracks in GaN films grown by combined hydride and metal organic vapor-phase epitaxial method.

Cracks appeared in GaN epitaxial layers which were grown by a novel method combining metal organic vapor-phase epitaxy (MOCVD) and hydride vapor-phase epitaxy (HVPE) in one chamber. The origin of cracks in a 22-µm thick GaN film was fully investigated by high-resolution X-ray diffraction (XRD), micro-Raman spectra, and scanning electron microscopy (SEM). Many cracks under the surface were first observed by SEM after etching for 10 min. By investigating the cross section of the sample with high-resolution micro-Raman spectra, the distribution of the stress along the depth was determined. From the interface of the film/substrate to the top surface of the film, several turnings were found. A large compressive stress existed at the interface. The stress went down as the detecting area was moved up from the interface to the overlayer, and it was maintained at a large value for a long depth area. Then it went down again, and it finally increased near the top surface. The cross-section of the film was observed after cleaving and etching for 2 min. It was found that the crystal quality of the healed part was nearly the same as the uncracked region. This indicated that cracking occurred in the growth, when the tensile stress accumulated and reached the critical value. Moreover, the cracks would heal because of high lateral growth rate.