[Development of a quality of life questionnaire for adults with anisometropic amblyopia].

Objective: To develop and validate the health-related quality of life (HRQOL) questionnaire for adult patients with anisometropic amblyopia. Methods: Cross-sectional study. A total of 170 adult patients with anisometropic amblyopia, 100 adult patients with other eye diseases and 80 healthy adults with normal vision were recruited at the First Affiliated Hospital of Nanjing Medical University, and 20 healthy adults with normal vision were recruited at Qinhuai Medical District of Easter Theater General Hospital of PLA from December 2019 to February 2020. Individual interviews of 30 adult patients with anisometropic amblyopia generated 80 questionnaire items. For item reduction, 40 adult patients with anisometropic amblyopia were asked to complete the 80-item questionnaire and responses were analyzed. Then factor analyses were performed to identify prominent factors (subscales). The reliability of the questionnaire was evaluated by Cronbach's α coefficient. The overall and sub-scale scores were the average scores of all included items, ranging from 0 (worst HRQOL) to 100 (best HRQOL). The final 20-item questionnaire was administered to additional 100 adult patients with anisometropic amblyopia, 100 adult patients with other eye diseases and 100 visually normal adults. Mean overall and subscale scores were compared across groups using one-way analysis of variance. Results: The final adult anisometropic amblyopia questionnaire (AAAQ) consisted of a function subscale and a psychosocial subscale, each containing 10 items. The Cronbach's α coefficients of the overall, function subscale and psychosocial subscale were 0.88, 0.78 and 0.78. There were 55 males and 45 females in 100 adult anisometropic amblyopia patients, with a median age of 26 years (range, 18 to 43 years). The age and gender distribution were matched with 100 adult patients with other eye diseases and 100 healthy adults with normal vision (all P>0.05). The mean overall score (28.63±9.18), function subscale score (27.69±9.88) and psychosocial subscale score (29.53±9.90) for adult patients with anisometropic amblyopia were significantly lower compared to adult patients with other eye diseases (71.28±8.14, P<0.01; 65.56±7.81, P<0.01; 76.85±10.76, P<0.01) and visually normal adults (84.54±9.13, P<0.01; 81.70±9.27, P<0.01; 87.38±10.06, P<0.01). Conclusion: The AAAQ meets the requirements for validity and reliability of a HRQOL questionnaire, and can be used to assess the HRQOL of adult patients with anisometropic amblyopia. (Chin J Ophthalmol, 2021, 57: 341-347).

GluR1 protects hypoxic ischemic brain damage via activating Akt signaling pathway in neonatal rats.

OBJECTIVE: To investigate the role of glutamic acid receptor 1 (GluR1) in hypoxic-ischemic brain damage (HIBD) in neonatal rats and its underlying mechanism. MATERIALS AND METHODS: 7-day-old neonatal rats received right common carotid artery (CCA) ligation for the establishment of HIBD. After the operation, rats were sacrificed at different time points (0, 4, 6, 12, 24, 48, and 72 h), respectively. Meanwhile, rats in Sham group underwent similar procedures without ligation. Lentivirus-GLUR1-shRNA (LV-GLUR1 shRNA group) was constructed and then transfected into the right lateral ventricles of rats to inhibit GluR1 in vivo. Rats received LV-control injection were selected in the control group (LV-control group). After injection of Lentivirus-GLUR1-shRNA, CCA ligation was performed in rats for HIBD construction. Western blot was performed to detect the protein levels of GLUR1, Akt, p-Akt, and vascular endothelial growth factor (VEGF) in brain tissues. Cell apoptosis was measured by TUNEL staining assay. RESULTS: After hypoxic ischemia (HI), GLUR1 expression increased gradually and reached a peak at 24 h. Meanwhile, p-Akt expression increased immediately and then gradually decreased. 24 h later, p-Akt expression increased again and peaked at 48 h. VEGF expression increased at 4 h after HI and reached a peak at 12 h. The expression levels of GLUR1, p-Akt, and VEGF in the brain tissues derived from rats transfected with LV-GLUR1 shRNA significantly decreased at both 4 h and 24 h after HI. In addition, results indicated that cell apoptosis was enhanced after LV-GLUR1 shRNA administration, suggesting the role of GLUR1 in protecting against HIBD. CONCLUSIONS: GLUR1 exhibits a remarkable protective role in HIBD, which may be related to the activation of the Akt signaling pathway and the upregulation of VEGF after HI.

Clinicopathologic analysis of 2736 salivary gland cases over a 11-year period in Southwest China.

OBJECTIVE: To investigate the epidemiological and clinicopathological characteristics of salivary gland tumors in southwest China in order to provide data for clinical diagnosis and other similar research. METHODS: Between March 2007 and December 2017, 2736 patients with salivary gland tumors were recruited, the clinical and pathological data were retrospectively analyzed. RESULTS: A total of 2736 patients had a ratio of males to females of about 1.02:1. The ratio of benign to malignant tumors was 3.46:1. Pleomorphic adenoma and adenoid cystic carcinoma had 50.8% and 7.2%, respectively. About 65.4% tumors occurred in the parotid gland. There was no significant difference between the tumor in the left or right parotid and the use of cell phones. There were significant differences between gender and both the characteristics and locations of salivary gland tumors (p < .05). There were also significant differences between the pathological characteristics and location of the salivary gland (p < .05). CONCLUSIONS: The salivary gland benign and malignant tumors were more common in pleomorphic adenoma and adenoid cystic carcinoma, most occurred in the parotid gland. The minor gland tumors are lower than other parts of China. The incidence of parotid gland tumors is not related to the use of cell phones.

Maxillary Reconstruction Using Submental Artery Island Flap and Sagittal Mandibular Ramus/Coronoid Process Graft Pedicled With Temporalis Muscle.

PURPOSE: The purpose of this study is to describe maxillary reconstruction using the submental artery island flap and the sagittal mandibular ramus and coronoid process graft pedicled with the temporalis muscle through the modified lateral lip and submandibular approach. MATERIALS AND METHODS: From May 2013 to September 2016, 11 patients with maxillary defects secondary to maxillary cancer ablation who underwent maxillary reconstruction using a submental artery island flap and a sagittal mandibular ramus and coronoid process graft pedicled with the temporalis muscle through the modified lateral lip and submandibular approach were enrolled in this prospective study. RESULTS: All submental artery island flaps and sagittal mandibular ramus and coronoid process grafts were successful, with satisfactory functional and esthetic outcomes. No functional impairment at the donor site occurred in any case. CONCLUSION: The submental artery island flap combined with the sagittal mandibular ramus and coronoid process graft is a feasible and acceptable technique for maxillary reconstruction in older patients because it is safe, quick, and straightforward to harvest and it offers a very acceptable esthetic and satisfactory outcome, with the advantage of low morbidity of the donor site. When combined with the 3-dimensional virtual operative method, the technique can improve postoperative outcomes.

[Clinical application and accuracy of the genioplasty surgical templates system for osseous genioplasty].

Objective: To develop and validate a new genioplasty templates system for monoblock osseous genioplasty. Methods: Thirty-six patients with chin deformities were enrolled in this study. The chin template system included a cutting guide and a repositioning guide for a genioplasty. Chin templates were designed in a computer and fabricated using a three-dimensional printing technique. The accuracy of the genioplasty templates were assessed by comparing the actual postoperative outcomes with the virtual plan. Results: All genioplasty was successfully completed by the template system. The largest linear root-mean-square deviation(RMSD) between the planned and the postoperative chin segments was 1.16 mm and the largest angular RMSD was 3.06°. Conclusions: The results showed that the chin template system provides a reliable method for transfer of genioplasty planning. The operation precision of the genioplasty can be improved by using the surgical templates system.

[Comparison between computer aided simulation and dental model orthognathic surgery for the treatment of patients with mandibular excess and facial asymmetries].

Objective: To evaluate the results of the orthognathic surgery with computer aided simulation-three-dimensional(3D) printed surgical guide and dental model surgery in the treatmemt of patients with mandibular excess and facial asymmetries. Methods: The coordinate system was built in ProPlan CMF 2.0 software, and the horizontal plane consisted of PoL, PoR, OrL, middle sagittal plane through nasion point and basion point and the plane perpendicular to the horizontal plane, coronoid plane through nasion point and the plane perpendicular to the horizontal plane and middle sagittal plane. The orientation of maxilla and mandibular distal segment was calculated by each triangle(U1-U6L-U6R, L1-L6L-L6R, Me-M5L-M5R) before and after orthognathic surgery. A total of 60 mandibular excess patients with facial asymmetries were enrolled in this retrospective study. They were divided into two groups, group Ⅰ with computer aided simulation, group Ⅱ with dental model surgery. The difference of maxillary occlusal plane roll and yaw angle, mandibular occlusal plane roll and yaw angle, and mandibular body plane roll and yaw angle were calculated. Statistical analysis was performed with SPSS 17.0 software. Results: The yaw angle of mandibular occlusal plane of the dental model surgery and computer aided simulation was 0.36°± 0.48° and 0.84° ± 0.36° (P=0.043), respectively. The roll angle of mandibular occlusal plane of the dental model surgery and computer aided simulation was 0.26°±0.79° and 0.54°±0.40°(P=0.032), respectively. The yaw angle of mandibular body plane of the dental model surgery and computer aided simulation was 0.60°± 1.04° and 0.23°±0.52°(P=0.008), respectively. The roll angle of mandibular body plane of the dental model surgery and computer aided simulation was 0.82° ± 0.72° and 0.53° ± 0.37° (P=0.028), respectively. The changes in computer aided simulation group were more obvious than that in the dental model surgery group, but the difference was not significant in the yaw angle of maxillary occlusal plane and the roll angle of maxillary occlusal plane between the two groups(P >0.05). Conclusions: It was more effective to correct mandibular asymmetry by computer aided simulation than dental model surgery.

PacBio SMRT assembly of a complex multi-replicon genome reveals chlorocatechol degradative operon in a region of genome plasticity.

We have sequenced a Burkholderia genome that contains multiple replicons and large repetitive elements that would make it inherently difficult to assemble by short read sequencing technologies. We illustrate how the integrated long read correction algorithms implemented through the PacBio Single Molecule Real-Time (SMRT) sequencing technology successfully provided a de novo assembly that is a reasonable estimate of both the gene content and genome organization without making any further modifications. This assembly is comparable to related organisms assembled by more labour intensive methods. Our assembled genome revealed regions of genome plasticity for further investigation, one of which harbours a chlorocatechol degradative operon highly homologous to those previously identified on globally ubiquitous plasmids. In an ideal world, this assembly would still require experimental validation to confirm gene order and copy number of repeated elements. However, we submit that particularly in instances where a polished genome is not the primary goal of the sequencing project, PacBio SMRT sequencing provides a financially viable option for generating a biologically relevant genome estimate that can be utilized by other researchers for comparative studies.

[The influence of urothelial carcinoembryonic antigen 1 on invasion and migration of oral carcinoma cell lines].

OBJECTIVE: To investigate the effects of long chain non-coding RNA urothelial carcinoembryonic antigen 1(UCA1) on invasion, migration and proliferation abilities in oral squamous cell carcinoma cell lines SCC15 and CAL27. METHODS: Small interfering RNA of UCA1(UCA1-siRNA) was transfected into SCC15 and CAL27 cell lines by Lipofectamine(TM) 3000 to silence UCA1 , and transfected negtive control si-RNA served as a control. The effect of UCA1-siRNA was detected by quantitative real time-polymerase chain reaction(qRT-PCR) to confirm the successful inhibition of UCA1 by siRNA. The matrix metalloproteinase 9(MMP-9) protein level was detected by Western blotting analysis. The effect of siRNA on cell proliferation and invasion was assessed by transwell migration assay and wound healing assay. Cell counting kit-8(CCK-8) assay was carried out to estimate the proliferation of two cell lines with different expression levels of UCA1. RESULTS: Expressions of UCA1 of SCC15 and CAL27 were successfully suppressed after transfected with siRNA which verified by qRT-PCR, and the efficiency of downregulation of SCC15 and CAL27 was 86.45%(P<0.001)and 78.24%(P<0.001), respectively. The migration, invasion and proliferation of SCC15 and CAL27 cell lines after transfected with siRNA were obviously restrained. The number of migration and invasion of CAL27 cells were 719.20±92.36 versus 208.00±25.58 (P=0.000 7) and 363.40 ± 45.96 versus 164.80 ± 24.68(P= 0.005 2), respectively, the number of migration and invasion of SCC15 cells were 437.20±54.75 vs 145.80±23.31(P=0.001 1) and 249.80±38.41 vs 63.80±11.11 (P=0.001 6), respectively (UCA1-si compare to negtive control), the relative proliferation rates of SCC15 and CAL27 were SCC15: R24 h=0.870, R48 h=0.863, R72 h=0.64, R96 h=0.732; CAL27: R24 h=0.913, R48 h=0.829, R72 h=0.756, R96 h= 0.705(P<0.05), and MMP-9 expression level was decreased by UCA1-siRNA compared with negative control. CONCLUSIONS: UCA1 could enhance the ability of invasion and migration of SCC15 and CAL27 cell lines via regulating MMP-9 protein expression, which suggests that UCA1 might play a pivotal role in oral squamous cell carcinoma invasion and progression.

Antitumor effect of VNP20009, an attenuated Salmonella, in murine tumor models.

VNP20009, a genetically modified strain of Salmonella typhimurium with deletions in the msbB and purI loci, exhibited antitumor activities when given systemically to tumor-bearing mice. VNP20009 inhibited the growth of subcutaneously implanted B16F10 murine melanoma, and the human tumor xenografts Lox, DLD-1, A549, WiDr, HTB177, and MDA-MB-231. A single intravenous injection of VNP20009, at doses ranging from 1 x 10(4) to 3 x 10(6) cfu/mouse, produced tumor growth inhibitions of 57-95%. Tumor volume doubling time, another indicator for tumor growth inhibition, also significantly increased in mice treated with VNP20009. Using mice with immune system deficiencies, we also demonstrated that the antitumor effects of VNP20009 did not depend on the presence of T and B cells. In addition, VNP20009, given intravenously, inhibited the growth of lung metastases in mice. Only live bacteria showed the antitumor effect.

An in vitro monitoring system for simulated thermal process in cryosurgery.

This paper describes a new in vitro experimental model that records temperature changes over a culture plate, which then can be used to assess the biological effects of cryosurgery. A cryoprobe and 16 thermocouples set up by a computer control system were used to monitor the temperature changes during freezing and thawing in a culture plate, and the data were used to create a temperature profile of the entire plate. Location of the thermocouples was confirmed by a digital camera viewing from under the plate, and temperature changes at any point in the interpolated areas were estimated using a curve fitting method. The estimated temperature was checked by sampling with four additional randomly placed thermocouples. Linear regression analysis showed that the estimated temperature and measured temperature were very close (correlation coefficients 0.98-0.99). MBT-2 tumor cells were cultured and then subjected to simulated cryosurgery. The surviving cells were stained with crystal violet and the cell death boundary was detected by image processing. Temperature history at the cell death boundary was retrieved and analyzed. With this system it is possible to recreate the temperature changes that result in a certain biological effect (such as cell death) during the process of simulated cryosurgery.

Lp approximation of Sigma-Pi neural networks.

A feedforward Sigma-Pi neural network with a single hidden layer of m neurons is given by mSigma(j=1) cjg (nPi(k=1) xk-thetak(j)/lambdak(j)) where cj, thetak(j), lambdak are elements of R. In this paper, we investigate the approximation of arbitrary functions f: Rn-->R by a Sigma-Pi neural network in the Lp norm. An Lp locally integrable function g(t) can approximate any given function, if and only if g(t) can not be written in the form Sigma(j=1)n Sigma(k=0)m alphajk(ln/t/)(j-1)tk.

Post-receptor signal transduction and regulation of 14(R),15(S)-epoxyeicosatrienoic acid (14,15-EET) binding in U-937 cells.

14(R),15(S)-epoxyeicosatrienoic acid (14,15-EET), a cytochrome P-450 monooxygenase (epoxygenase) metabolite of arachidonic acid has been reported to induce adhesion of a monocyte cell line (U-937) to cultured endothelial cells. In this study, we identified a population of specific, high affinity binding sites for 14(R),15(S)-EET in U-937 cell surface with Kd of 13.84 +/- 2.58 nM and Bmax of 3.54 +/- 0.28 pmol/10(6) cells. The specific binding of [3H]-14,15-EET on U-937 cells is more effectively displaced by 14(R),15(S)-EET than the 14(S),15(R)-isomer thus indicating stereospecificity. The binding was sensitive to various protease treatments suggesting the binding site is protein in nature. 14,15-EET binding in U937 cells is attenuated by cholera toxin (CT) and dibutyryl cAMP. Mean binding site density (Bmax) decreased 31.61% and 34.8% by the pretreatment with cholera toxin (200 micrograms/ml) and dibutyryl cAMP (300 nM), respectively, without affecting the dissociation constant. Under similar conditions, pertussis toxin (20-200 ng/ml) was less effective as compared to CT and dibutyryl cAMP. The down regulation of 14,15-EET binding caused by dibutyryl cAMP in U-937 cell was reversed by a specific protein kinase A (PKA) inhibitor, H-89, but not by the PKC inhibitor K252a. Thus, the results suggest that the specific binding site of 14,15-EET in U-937 cells is associated with a receptor that could be down regulated through an increase in intracellular cAMP and activation of a PKA signal transduction mechanism. We propose that the signal transduction mechanism of 14,15-EET begins with the binding of the receptor, which leads to the increase of intracellular cAMP levels and the activation of PKA, and finally with the down regulation of 14,15-EET receptor binding.

Natural inactivation of a common HLA allele (A*2402) has occurred on at least three separate occasions.

HLA-A*2402 is common and widely distributed in human populations. Several individuals were identified who type genotypically for A*2402, but are serologically null for the HLA-A24 Ag. Sequencing and transfection of genomic DNA fragments containing null and wild-type A*2402 alleles, and the related A*2301 allele, revealed three different null alleles (A*2409N, A*2411N, and A*2402(low)), each of which differs from A*2402 by a single nucleotide change within the 6.7-kb sequence. The A*2301 and A*2402 sequences differ by no substitutions additional to those previously determined for the 1.1-kb cDNA. In exon 4, A*2409N has an in-frame stop codon, while A*2411N has a nucleotide insertion that alters the reading frame, causing premature termination. A*2402(low) has a nucleotide substitution near the splice acceptor site for intron 2 that impairs the production of correctly spliced mRNA. For A*2409N and A*2411N, mRNA is undetectable by Northern analysis, whereas A*2402(low) produces a low level of mRNA and a concomitant amount of normal A*2402 protein at the cell surface. The protein expressed from the A*2402(low) allele is sufficient to stimulate an alloreactive T cell response. On a background of unexpected sequence homogeneity, the single nucleotide changes in the A*2409N, A*2411, and A*2402(low) alleles have dramatic effects upon gene expression and are of likely importance for HLA matching in clinical transplantation. Segregation of at least three independently inactivated A*2402 alleles in human populations raises the possibility that loss of A*2402 may be the result of natural selection.

[A liveborn infant with triploidy (69, XXX): report of one case].

Triploidy is not rare and present in about 1% of all recognized human pregnancies, although most of these pregnancies end in spontaneous abortion during the first trimester. Survival of the fetus up to 20 weeks or beyond is rare. Therefore, liveborn infants with triploidy are very rare. Here is a report on a female liveborn infant with triploidy (69,XXX), who was born to a 27-year-old healthy mother. The clinical features are growth retardation, head-to-body disproportion, wide posterior fontanelle, hypertelorism, micrognathia, bilateral pre-auricular polyps, syndactyly of left 3rd and 4th fingers, syndactyly of right 2nd and 3rd fingers and talipes equinovarus. The infant died 4 hours after birth. The autopsy revealed transposition of great vessels, ventricular septal defect, one lobe of left lung and 2 lobes of right lung and duodenal atresia.

[Low-dose recombinant human tumor necrosis-alpha in HL-60 leukemic cell line: effect on induced differentiation and regulating expression of c-myc, c-fos proto-oncogenes].

The promyelocytic leukemic cell line HL-60 with 20-200U/ml of low-dose rhTNF-a cultured in liquid culture system in vitro was used to observe the effect on HL-60 by TNF. TNF within dose of 50-200 U/ml can induce HL-60 cell differentiation along the monocytic-macrophage pathway, and inhibit HL-60 cell proliferation. The total RNA of HL-60 cell was used to hybrize to v-myc or v-fos probe by dot blot. We detected the expression changes of c-myc or c-fos proto-oncogene by 1-100U/ml of TNF inducing HL-60 cell for 1-12 hours. TNF could regulate the level of c-myc or c-fos mRNA, the transcription of c-myc was inhibited remarkdly, and the expression of c-fos was increased early. The results indicated that TNF in low-dose have effect on inducing HL-60 cell differentiation and regulating expression of multioncogene.

[The effects of recombinant human transforming growth factor beta on inducing differentiation of fresh leukemia cells in acute monocytic leukemia M5].

By using fresh leukemia cells from 5 cases of acute monocytic leukemia M5 as in vitro model, we investigated the effects of recombinant human transforming growth factor beta 1 (rhTGF-beta 1) on differentiation induction of fresh leukemia cells. The results indicated that after 6 days of induction with TGF-beta 1 in a concentration of 10 ng/ml, leukemia cells in 5 AML-M5 patients differentiated obviously to maturation. The proportion of monoblasts and premonocytes was reduced, while that of mature mononuclear cells elevated. Following administration of TGF-beta 1, alpha-nonspecific esterase (alpha-NSE), whose expression could be inhibited by sodium fluoride, remained positive and peroxidase (POX) was shown to be weakly positive. These results demonstrated that TGF-beta 1 may induce in vitro differentiation of fresh leukaemia cells, but the reactions to TGF-beta 1 may vary in different cases.

[Inhibition of proto-oncogene c-myc expression with recombinant human tumor necrosis factor-alpha and interferon-gamma in HL-60 cell lines and acute myelocytic leukemic fresh cells].

HL-60 cell lines and AML fresh bone marrow cells were incubated with rhTNF-alpha and rhIFN-gamma in suspension culture system. Then total RNA was prepared for dot blot with 32P nick-translated c-myc DNA probe. The expression changes of c-myc oncogene when the HL-60 cell lines were treated with rhTNF-alpha and IFN-gamma and the AML fresh bone marrow cells treated with rhTNF-alpha alone were observed. The results showed that when the HL-60 cells were treated with 100U/ml or 500U/ml IFN-gamma and 50U/ml rhTNF-alpha for 8 hours, the expression of c-myc oncogene can be inhibited remarkably. The combination of rhIFN-gamma and rhTNF-alpha shows synergistic effect on inhibition of c-myc expression. High expression of c-myc was found in 8 patients with AML; c-myc mRNA level decreased remarkably after treatment of fresh bone marrow cells with 50U/ml rhTNF-alpha for 12 hours in 6 cases, while the remaining 2 cases showed minimal changes. The results demonstrate that rhTNF-alpha have inhibitive effect on c-myc expression in HL-60 cells and AML fresh leukemic cells. It also indicates the possibility of treating AML with low-dose rhTNF-alpha.