OBJECTIVES: This study aimed to explore the expression of lysosomal-associated membrane protein 5 (LAMP5) and microRNA (miR)-302a-3p in oral squamous cell carcinoma (OSCC) and their functional mechanism on the invasion and metastasis of OSCC. METHODS: The expression of LAMP5 in OSCC and its sensitivity as a prognostic indicator were analyzed on the basis of The Cancer Genome Atlas database. Western blot, quantitative reverse transcription polymerase chain reaction, and cell immunocytochemistry were used to detect the expression of LAMP5 in OSCC tissues and cells. The effect of LAMP5 on the proliferation, migration, and invasion of OSCC cells was evaluated through cell counting kit-8, immunocytochemistry, migration, and invasion assays, respectively. The miRNA targeting prediction websites were used to predict the miR that regulates LAMP5 and verify the targeted regulatory effect of miR-302a-3p on LAMP5. The effect of LAMP5 knockdown on OSCC tumor growth was evaluated in a nude mouse tumorigenesis model. RESULTS: LAMP5 was highly expressed in OSCC tissues and cells. It showed high sensitivity in the early diagnosis of OSCC. LAMP5 knockdown significantly inhibited the proliferation, migration, and invasion of OSCC cells, whereas LAMP5 overexpression increased these cell activities. The expression of LAMP5 was regulated by miR-302a-3p. In vivo, LAMP5 knockdown significantly inhibited the growth of OSCC tumor. CONCLUSIONS LAMP5 promotes the malignant progression of OSCC by enhancing the proliferation, migration, and invasion of OSCC cells. The expression of LAMP5 is negatively regulated by miR-302a-3p.
MicroRNAs/metabolism* ; Mouth Neoplasms/metabolism* ; Humans ; Animals ; Carcinoma, Squamous Cell/genetics* ; Neoplasm Invasiveness ; Cell Proliferation ; Mice, Nude ; Cell Movement ; Lysosomal Membrane Proteins/genetics* ; Mice ; Cell Line, Tumor ; Neoplasm Metastasis

OBJECTIVE: To assess the value of chromosomal microarray analysis (CMA) and trio-whole exome sequencing (trio-WES) for fetuses with increased nuchal translucency (NT) thickness. METHODS: Sixty two pregnant women who had visited Urumqi Maternal and Child Care Health Hospital between June 2018 and June 2020 for NT ≥ 3.0 mm at 11 ~ 13⁺⁶ gestational weeks were selected as study subjects. Relevant clinical data were collected. The patients were divided into 3.0 ~ ＜3.5 mm (n = 33) and ≥3.5 mm groups (n = 29). Chromosome karyotyping analysis and chromosomal microarray analysis were carried out. And trio-WES analysis was performed on 15 samples with NT thickening but negative CMA results. The distribution and incidence of chromosomal abnormalities in the two groups were compared by using chi-square test. RESULTS: The median age of the pregnant women was 29 years old (22 ~ 41 years old), the median thickness of NT was 3.4 mm (3.0 ~ 9.1 mm), and the median gestational age at the detection was 13⁺⁴ weeks (11⁺⁵ ~ 13⁺⁶ weeks). Chromosome karyotyping analysis has detected 12 cases of aneuploidies and 1 case of derivative chromosome. The detection rate was 20.97% (13/62). CMA has detected 12 cases of aneuploidies, 1 case of pathogenic CNV and 5 cases of variant of uncertain significance (VUS), with a detection rate of 29.03% (18/62). The aneuploidy rate for the NT ≥ 3.5 mm group was higher than that for the 3.0 ≤ NT < 3.5 mm group [3.03% (1/33) vs. 41.38% (12/29), χ² = 13.698, P < 0.001]. There was no statistically significant difference between the two groups in the detection rate of fetal pathogenic CNV and VUS (χ² = 0.028, P > 0.05). Trio-WES analysis of 15 samples with negative CMA result and no structural abnormality has identified 6 heterozygous variants, including SOS1: c.3542C>T (p.A1181V) and c.3817C>G (p.L1273V), COL2A1: c.436C>T (p.P146S) and c.3700G>A (p.D1234N), LZTR1: c.1496T>C (p.V499A), and BRAF: c.64G>A (p.D22N), respectively. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), all of the variants were rated as VUS. CONCLUSION NT thickening can indicate chromosome abnormality, and CMA and trio-WES may be used for the prenatal diagnosis.
Pregnancy ; Humans ; Female ; Adult ; Infant ; Nuchal Translucency Measurement/methods* ; Prenatal Diagnosis/methods* ; Chromosome Aberrations ; Aneuploidy ; Fetus/diagnostic imaging* ; Ultrasonography, Prenatal ; DNA Copy Number Variations ; Transcription Factors

Objective:To determine the expression of silent information regulator 1 (Sirt1) , Sirt3 and hypoxia-inducible factor 1α (HIF-1α) in cutaneous squamous cell carcinoma (CSCC) tissues and cells, and to explore their role in the occurrence and development of CSCC.Methods:From January 2019 to December 2020, 30 lesional skin tissues were obtained from patients with histopathologically confirmed poorly-, moderately- or well-differentiated CSCC, and 30 normal skin tissues were obtained from patients with non-cancerous diseases in Department of Dermatology, General Hospital of Ningxia Medical University. A CSCC cell line A431 and a human keratinocyte cell line HaCaT were cultured. Immunohistochemical study, Western blot analysis and real-time quantitative PCR (RT-PCR) were performed to determine the protein and mRNA expression of Sirt1, Sirt3 and HIF-1α in CSCC tissues of different grades of differentiation and normal skin tissues, cytochemical and immunofluorescence staining and RT-PCR were conducted to determine the protein and mRNA expression of Sirt1, Sirt3 and HIF-1α in A431 and HaCaT cells, respectively. Comparisons of measurement data among multiple groups were performed by using one-way analysis of variance, and comparisons between two groups by using t test. Results:Immunohistochemical study showed that the expression level of Sirt3 (expressed as the average optical density) was 100 ± 12.12, 117.72 ± 26.23, 127.32 ± 24.45, 132.71 ± 31.61 in the normal skin tissues and well-, moderately- and poorly-differentiated CSCC tissues respectively, and there was a significant difference among these groups ( F = 20.14, P < 0.001) ; the expression of Sirt1 and HIF-1α increased in turn from the normal skin tissues to the well-, moderately- and poorly-differentiated CSCC tissues, and significantly differred in these groups ( F = 174.50, 225.00, respectively, both P < 0.001) . As Western blot analysis revealed, the expression level of Sirt3 significantly differed among the normal skin tissues, well-, moderately- and poorly-differentiated CSCC tissues (expressed as relative gray value: 1.000 ± 0.132, 1.403 ± 0.411, 1.387 ± 0.393, 1.677 ± 0.683, respectively; F = 34.97, P < 0.001) , and so did the expression levels of Sirt1 and HIF-1α ( F = 69.29, 199.90, respectively, both P < 0.00l) , with a gradually increasing trend in their expression levels from the the normal skin tissues to well-, moderately- and poorly-differentiated CSCC tissues. RT-PCR showed that the mRNA expression of Sirt3, Sirt1 and HIF-1α was sequentially increased from the normal skin tissues to well-, moderately- and poorly-differentiated CSCC tissues, and significant differences were observed among these groups ( F = 113.00, 174.50, 50.33, respectively, all P < 0.001) . The protein expression levels of Sirt3, Sirt1 and HIF-1α were significantly higher in the A431 cells than in the HaCaT cells ( t = 16.75, 18.34, 27.76, respectively, all P < 0.001) , and so were their mRNA expression levels ( t= 14.22, 9.62, 16.86, respectively, all P < 0.001) . Conclusion:Increased expression of Sirt3, Sirt1 and HIF-1α was observed in CSCC tissues and cells, which may promote the occurrence and development of CSCC.

Objective To analyze the incidence and mortality of male breast cancer in Hubei Province from 2012 to 2015. Methods The data reported by the Hubei Provincial Cancer Registry from 2012 to 2015 were collected for analyzing the incidence and mortality trends of male breast cancer according to year, urban and rural areas and age. We also compared the difference of incidence and mortality between male and female breast cancer. Results A total of 106 new cases of male breast cancer were reported in Hubei Cancer Registry from 2012 to 2015 with an incidence rate of 0.43/10⁵, and 10052 new cases of female breast cancer with an incidence rate of 42.76/10⁵; the male-to-female incidence ratio was 1:99. Male breast cancer death cases was 37 with the death rate of 0.15/10⁵, and female breast cancer death cases was 2201 with the death rate of 9.36/10⁵; the male-to-female mortality ratio was 1:62. The incidence and mortality of breast cancer between male and female varied by year, urban and rural areas. Male breast cancer was less common in young men (< 35 years old), and the incidence increased with age after 35 years old; male breast cancer deaths were rare in men under 45 years old, and the mortality rate increased with age. The incidence and mortality of male breast cancer patients over 85 years old reached the peak. Conclusion Male breast cancer is rare in clinical practice, and the incidence rate is much lower than that of female breast cancer. Even so, it is necessary to study the epidemiological law of male breast cancers' incidence and mortality, which can be used for prevention and treatment of male breast cancer.

Objective To detect mutations in the ARAD1 gene in a pedigree with dyschromatosis symmetrica hereditaria (DSH).Methods Genomic DNA was extracted from the peripheral blood of 8 family members (including 5 patients with DSH and 3 unaffected members) in the pedigree with DSH,as well as 100 unrelated healthy controls.All the 15 exon sequences of the ADAR1 gene were amplified by polymerase chain reaction (PCR)followed by direct sequencing.Then,mutations were detected in comparison with the standard sequence of the ADAR1 gene in Genebank.Results A nonsense mutation C.1420C ＞ T (p.Arg474X) was identified at position 1 420 in exon 2 of the ADAR1 gene in the 5 patients with DSH,but not in the 3 unaffected members or 100 unrelated healthy controls.Conclusion The nonsense mutation C.1420C ＞ T in the ADAR1 gene is the causative mutation in the pedigree with DSH.

Objective To study the impacts of different tube voltage and different noise index(NI) guided automatic tube current modulation on the image quality and radiation dose in cerebrovascular imaging and determine the optimal scanning condition. Methods PH3 angiographic CT head phantom was used for head CTA examination. Scanning protocols: all the scanning objectives were divided into three groups according to the different tube voltages of 80, 100 and 120 kV. Each group applied certain tube current(300,400 mA)and automatic tube current modulation technique with NI from 3 to 10 to perform head CTA. There were 30 scanning proposals with different parameter combinations of tube voltage and tube current. The radiation dose [ CT dose index volume (CTDIvol), dose length product (DLP) and effective dose (ED)], objective indicators of images(CT value of the blood vessels and its noise, CT value of brain tissue and its noise, signal-to-noise ratio , contrast-to-noise ratio) and the subjective scores of the five cerebrovascular segments were recorded. Differences of CT value of the blood vessels and its noise, CT value of brain tissue and its noise, signal-to-noise ratio and contrast-to-noise ratio between different tube voltages and tube currents were analyzed by One-Way ANOVA. Results When the tube voltage was certain, the CTDIvol, DLP and ED were all dropped while NI was increased from 3 to 10. Compared with group(120 kV, 300 mA), CTDIvol of group with 100 kV, 300 mA decreased 35.32%(12.22/34.59), CTDIvol of group(100 kV, NI=6) decreased 46.72%(16.16/34.59). Compared with group(100 kV, 300 mA), CTDIvol of group (100 kV, NI=6) decreased 17.61%(3.94/22.37). When the tube voltage was certain and the tube current and NI were not certain, there is no statistical difference (P>0.05) between CT values of blood vessel and brain, while blood vessel noise, noise of brain, SNR and CNR showed statistical difference (P<0.05). When tube current and NI were certain while tube voltage was varied, all objective indicators discussed above all exhibited statistical difference (P<0.05). SNR and CNR of group(100 kV, NI=6) were higher than group(120 kV, 300 mA) with 6.31%(2.69/42.66)and 7.18%(2.64/36.78), respectively. The tube voltage, NI and tube current had no effect on the subjective scores of first and second grade vessel but greater impact on the fourth and fifth grade vessel. Conclusion In the head CTA scanning, combined the use of NI 6 guided automatic adjustment tube current and low tube voltage(100 kV)technique not only can get better image quality but also significantly decreased the radiation dose.

Objective To study the impacts of different tube voltage and different noise index(NI) guided automatic tube current modulation on the image quality and radiation dose in cerebrovascular imaging and determine the optimal scanning condition. Methods PH3 angiographic CT head phantom was used for head CTA examination. Scanning protocols: all the scanning objectives were divided into three groups according to the different tube voltages of 80, 100 and 120 kV. Each group applied certain tube current(300,400 mA)and automatic tube current modulation technique with NI from 3 to 10 to perform head CTA. There were 30 scanning proposals with different parameter combinations of tube voltage and tube current. The radiation dose [ CT dose index volume (CTDIvol), dose length product (DLP) and effective dose (ED)], objective indicators of images(CT value of the blood vessels and its noise, CT value of brain tissue and its noise, signal-to-noise ratio , contrast-to-noise ratio) and the subjective scores of the five cerebrovascular segments were recorded. Differences of CT value of the blood vessels and its noise, CT value of brain tissue and its noise, signal-to-noise ratio and contrast-to-noise ratio between different tube voltages and tube currents were analyzed by One-Way ANOVA. Results When the tube voltage was certain, the CTDIvol, DLP and ED were all dropped while NI was increased from 3 to 10. Compared with group(120 kV, 300 mA), CTDIvol of group with 100 kV, 300 mA decreased 35.32%(12.22/34.59), CTDIvol of group(100 kV, NI=6) decreased 46.72%(16.16/34.59). Compared with group(100 kV, 300 mA), CTDIvol of group (100 kV, NI=6) decreased 17.61%(3.94/22.37). When the tube voltage was certain and the tube current and NI were not certain, there is no statistical difference (P>0.05) between CT values of blood vessel and brain, while blood vessel noise, noise of brain, SNR and CNR showed statistical difference (P<0.05). When tube current and NI were certain while tube voltage was varied, all objective indicators discussed above all exhibited statistical difference (P<0.05). SNR and CNR of group(100 kV, NI=6) were higher than group(120 kV, 300 mA) with 6.31%(2.69/42.66)and 7.18%(2.64/36.78), respectively. The tube voltage, NI and tube current had no effect on the subjective scores of first and second grade vessel but greater impact on the fourth and fifth grade vessel. Conclusion In the head CTA scanning, combined the use of NI 6 guided automatic adjustment tube current and low tube voltage(100 kV)technique not only can get better image quality but also significantly decreased the radiation dose.

Objective: To investigate Oct4 and Sox2 protein expression in tongue squamous cell carcinoma (TSCC) and the relationships between the expressions of Oct4 and Sox2 and clinical pathological characteristics and survival of patients. Methods: The paraffin imbedded tissue specimens of 51 patients with histologically confirmed TSCC were included. Immunohistochemistry was employed to detect the protein expression of Oct4 and Sox2 in 51 TSCC tissue samples. The protein expression levels of Oct4 and Sox2 and their relationships with both clinicopathological features and survival of patients with TSCC were evaluated. Results: In 51 TSCC cases,positive expressions of Oct4 and Sox2 were mainly located in the nucleus of tumor cells. The expression of Oct4 was strongly positive in 27 cases (53%), weakly positive in 16 (31%) and negative in 8 (16%), whereas that of Sox2 was strongly positive in 25 cases (49%), weakly positive in 22 (43%) and negative in 4 (8%). Oct4 and Sox2 expression levels were significantly correlated with the histological grade of TSCC (P=0.004, P=0.006, respectively), not correlated with age, gender, T stage, tobacco smoking and alcohol consumption status (P>0.05), but Oct4 expression level was significantly associated with lymph node metastasis (P=0.001). Sox2 expression level was not associated with lymph node metastasis (P>0.05). The expression of Sox2 was significantly correlated with Oct4 (P<0.001). Oct4 and Sox2 expression was associated with poor overall survival of patients with TSCC (P=0.001, P=0.002, respectively), cases with higher Oct4 and Sox2 expression had the poorest overall survival (P<0.001). Sox2 expression and lymph node metastasis were independent prognostic factors of overall survival in patients with TSCC (P=0.02, P=0.001, respectively). Conclusions Sox2 had independent prognostic effects on overall survival, suggesting that Sox2 expression may be an usefull indicator for predicting the prognosis of patients with TSCC.

Objective: To investigate the efficacy and safety of IA regimen which contains idarubicin (IDA) 8 mg/m², 10 mg/m² or 12 mg/m² as induction chemotherapy for adult patients with de-novo acute myeloid leukemia (AML) . Methods: A total of 1 215 newly diagnosed adult AML patients, ranging from May 2011 to March 2015 in the First Affiliated Hospital of Soochow University and other 36 clinical blood centers in China were enrolled in the multicenter, single-blind, non-randomized, clinical controlled study. To compare the response rate of complete remission (CR) , adverse events between different dose idarubicin combined with cytarabine (100 mg/m²) as induction chemotherapy in newly diagnosed patients of adult AML. Results: Of 1 207 evaluable AML patients were assigned to this analysis of CR rate. The CR rates of IDA 8 mg/m² group, IDA 10 mg/m² group and IDA 12 mg/m² group were 73.6% (215/292) , 84.1% (662/787) and 86.7% (111/128) , respectively (P<0.001) . After adjusted for age, blast ratio of bone marrow, FAB classification and risk stratification, the odds ratios (95% CI) of IDA 10 mg/m² group and IDA 12 mg/m² group were 0.49 (0.34-0.70) and 0.36 (0.18-0.71) , as compared with the IDA 8 mg/m² group (P<0.001, P=0.003) . In the intermediate and favorable groups, CR rates was 76.5% (163/213) , 86.9% (506/582) and 86.1% (68/79) in different doses of IDA (P=0.007) . Interestingly, IA regimen with IDA 10 mg/m² was the only beneficial factor affecting CR in this group after adjusted for age, blast ratio of bone marrow and FAB classification[OR=0.47 (95% CI 0.31-0.71) , P<0.001]. CR rates in adverse group was 50.0% (18/36) , 60.6% (43/71) and 81.8% (18/22) respectively (P=0.089) . However, the odds ratios (95% CI) of IDA 12 mg/m² when compared with the IDA 8 mg/m² was 0.22 (0.06-0.80) , after adjusted for age, blast ratio of bone marrow and FAB classification. The median time (days) of neutrophil count less than 0.5×10⁹/L in IDA 8 mg/m² group, IDA 10 mg/m² group and IDA 12 mg/m² group were 14 (11-18) , 15 (11-20) and 18 (14-22) , respectively (P=0.012) and of platelet count lower than 20×10⁹/L were 14 (7-17) , 15 (11-20) and 17 (15-21) , respectively (P=0.001) . The incidences of lung infection in the three groups were 9.8%, 13.5% and 25.2%, respectively (P<0.001) . Conclusions For young adult patients (aged 18-60 years) with AML in China, intensifying induction therapy with idarubicin 10 mg/m² is clinically superior to IDA 8 mg/m² and IDA 12 mg/m² in favorable intermediate AML subgroup. However, idarubicin 12 mg/m² is more suitable to adverse AML subgroup.