Application of next-generation sequencing in detection of BRCA1/2 and homologous recombination repair pathway multi-genes germline mutation and correlation analysis / 中华预防医学杂志

Objective: To investigate the germline mutation status of related genes in breast cancer patients and high-risk individuals by next-generation sequencing. To analyze the correlations between homologous recombination repair (HR) pathway gene mutation status and clinicopathological characteristics of breast cancer patients. To supplement the database of breast cancer related gene mutations in Chinese population. Methods: This study is a cross-sectional study. From October 2020 to September 2021, whole blood samples were collected from 350 breast cancer patients and 49 high-risk individuals, admitted to Peking University People's Hospital and accepted genetic testing voluntarily. Germline mutations in 32 breast cancer related genes were detected by NGS. The clinicopathological characteristics, including age at the onset, family history, unilateral/bilateral tumor, Luminal typing (Luminal A subtype, Luminal B subtype, HER2-enriched subtype and triple negative breast cancer), tumor size and metastasis, were analyzed, and the correlations between HR pathway gene mutation status and clinicopathological characteristics were analyzed by Chi-squared test and Fisher's exact probability test. Results: Among 350 breast cancer patients, 64 (18.3%) cases carried gene pathogenic mutations (including pathogenic and likely pathogenic mutations), including 47 (13.4%) in BRCA1/2, 16 (4.6%) in non-BRCA1/2 genes, 1 (0.3%) in BRCA2 and FANCL. Among 49 high-risk individuals, 7 (14.3%) cases carried gene pathogenic mutations, including 6 (12.3%) in BRCA1/2 and 1 (2%) in ATM genes. BRCA1/2 pathogenic mutations were associated with age at the onset (18%, 8.7%, χ²=6.346, P=0.012), and the BRCA1/2 pathogenic mutation frequency was higher in patients diagnosed at age ≤45 years. HR pathway gene mutations (including pathogenic, likely pathogenic and uncertain significance mutations) were correlated with unilateral/bilateral tumor (49.5%, 68.4%, χ²=4.841, P=0.028) and Luminal typing (45.7%, 62.2%, 32%, 60%, χ²=12.004, P=0.007), and the HR mutation frequencies were higher in patients with bilateral tumor, Luminal B breast cancer and triple negative breast cancer (TNBC). Conclusion: The BRCA1/2 pathogenic mutation frequency in high-risk individuals is similar to that in breast cancer patients, and BRCA1/2 testing is helpful to guide breast cancer screening and prevention in high-risk individuals. Patients with early onset breast cancer, bilateral breast cancer, Luminal B breast cancer and TNBC have higher mutation frequencies of HR pathway genes, and HR pathway genes testing should be conducted as soon as possible to provide laboratory evidence for diagnosis, treatment, prognosis and risk evaluation of breast cancer.

Clinic analysis of fundus screening and the risk factors of retinal hemorrhage in neonates / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To understand the basic conditions of neonatal fundus lesions and analyze the influencing factors of retinal hemorrhage.<p>METHODS: Totally 289 neonates from our hospital in the department of obstetric ward and neonatal intensive care unit from January 1, 2016 to May 31, 2018 were screened for fundus oculi disease by Retcam Ⅲ. The following information were collected such as gender, gestational age, history of asphyxia, maternal pregnancy and delivery way.<p>RESULTS: Among the 289 infants, there were 97 premature infants and 192 term infants. Sixty-four(22.1%)were detected with fundus lesions. Among which, 34(11.8%)were detected with retinal hemorrhage and 23(8.0%)retinopathy of prematurity. Single factor analysis showed that the rate of retinal hemorrhage in premature infants was higher than term(<i>P</i><0.05). The rate of retinal hemorrhage in neonates with perineal delivery was higher than cesarean section(<i>P</i><0.05). The rate of retinal hemorrhage in neonates with a history of asphyxiation was higher than that without(<i>P</i><0.05). The rate of retinal hemorrhage in neonates with high blood pressure during pregnancy was higher than that without(<i>P</i><0.05). After adjusting for the effects of risk factors, preterm infants, perineal births, history of asphyxiation, and high blood pressure during pregnancy were the high risk factors for neonatal retinal hemorrhage(<i>P</i><0.05).<p>CONCLUSION: The incidence of neonatal retinal hemorrhage is relatively high, which is related to factors such as gestational age, mode of delivery, history of asphyxia and history of maternal pregnancy hypertension.

Effect of Metformin on Proliferation Capacity, Apoptosis and Glycolysis in K562 Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of metformin on the proliferation, apoptosis and energy metabolism of acute myeloid leukemia (AML) K562 cells and the possible mechanism.@*METHODS@#Different doses (0, 5, 10, 20 and 30 mmol/L) of metformin was added into the K562 cells, which were cultivated for 24 h, 48 h and 72 h. The inverted optical microscope was used to observe the cell growth, CCK 8 was used to detect the cell vitality. The appropriate metformin doses (0, 10, 20 and 30 mmol/L) and the best time (48 h) were selected for subsequent experiments. The flow cytometer with Annexin V-FITC /PI doulde staining was used to detect apoptosis; the glucose detection kit and lactate detection kit were used to detect glucose consumption and lactate production; fluorescence quantitative PCR was used to detect glycolysis-related gene expression, and Western blot was used to detect protein expression.@*RESULTS@#Metformin inhibited the proliferation of K562 cells in a dose-dependent manner (r=0.92), and the relative survival in the 30 mmol/L group was as low as 19.84% at 72 h. When treated with metformin for 48 h, the apoptosis rates of 0, 10, 20 and 30 mmol/L groups were 5.14%, 12.19%, 26.29% and 35.5%, respectively. Compared with the control group, the glucose consumption and lactate secretion of K562 cells treated with metformin were significantly reduced (P<0.05), and showed a dose-dependent effect(r=0.94,r=0.93,respectively). Metformin inhibited the expression of GLUT1, LDHA, ALDOA, PDK1, and PGK1 genes of K562 cells (P<0.05) showing a dose-dependent manner(r=0.83，r=0.80，r=0.72，r=0.76，r=0.73,respectively). Metformin inhibited the expression of P-Akt, P-S6, GLUT1, LDHA proteins of K562 cells(P<0.05), showing a dose-dependent relationship(r=0.80，r=0.92，r=0.83，r=0.92,respectively).@*CONCLUSION@#Metformin can inhibit the growth and proliferation of K562 cells and promote the apoptosis of K562 cells by inhibiting glycolysis energy metabolism. PI3K/Akt/mTOR signaling pathway may be one of the molecular mechanisms of metformin on k562 cells.

Study on the quality of life and influencing factors in patients with diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

·AIM: To research the quality of life ( QOL ) and influencing factors in patients with diabetic retinopathy (DR). ·METHODS: A total of 103 diagnosed case of diabetic retinopathy from January 2017 to August 2017 in Daping Hospital of Chongqing were enrolled in this study. The questionnaire survey was conducted using Scale of Quality of Life in Patients with Visual Impairment (SQQL-VI ) and social support revalued scale. The factors influencing their quality of life were also analyzed. The data were analyzed by SPSS 25. 0 software. Proportions were compared by using the chi-square test and the means were compared by using the t-test. The factors of DR such as age, gender, education and social support were analyzed using stepwise multiple linear regression. ·RESULTS: The scores in the patients with diabetic retinopathy included total quality of life 63. 59 ± 9. 58, symptoms and visual function 53. 13 ± 8. 51, body function 28. 39 ± 3. 86, social activities 27. 95 ± 3. 63, psychological aspects 27. 78 ± 3. 85. The single factor analysis showed that there were significant differences in age, education level occupation, monthly income, the style of medical cost, course of disease, and social support (P<0. 05). Multivariate linear regression indicated that age, education level, the style of medical cost, course of disease, and social support were statistically significantly associated with the quality of life (P<0. 05). ·CONCLUSION: To improve the quality of life of DR patients, it is essential to reduce the financial burden and increase the social support.

The Blood Oxygenation T₂* Values of Resectable Esophageal Squamous Cell Carcinomas as Measured by 3T Magnetic Resonance Imaging: Association with Tumor Stage

OBJECTIVE: To explore the association between the blood oxygenation T₂* values of resectable esophageal squamous cell carcinomas (ESCCs) and tumor stages. MATERIALS AND METHODS: This study included 48 ESCC patients and 20 healthy participants who had undergone esophageal T₂*-weighted imaging to obtain T₂* values of the tumors and normal esophagi. ESCC patients underwent surgical resections less than one week after imaging. Statistical analyses were performed to identify the association between T₂* values of ESCCs and tumor stages. RESULTS: One-way ANOVA and Student-Newman-Keuls tests revealed that the T₂* value could differentiate stage T1 ESCCs (17.7 ± 3.3 ms) from stage T2 and T3 tumors (24.6 ± 2.7 ms and 27.8 ± 5.6 ms, respectively; all p(s) 0.05) or between N stages (N1 vs. N2 vs. N3: 24.7 ± 6.9 ms vs. 25.4 ± 4.5 ms vs. 26.8 ± 3.9 ms, respectively; all p(s) > 0.05). The former tests illustrated that the T₂* value could differentiate anatomic stages I and II (18.8 ± 4.8 ms and 26.9 ± 5.9 ms, respectively) or stages I and III (27.3 ± 3.6 ms). ROC analysis depicted the same cutoff T₂* value of 21.3 ms for either differentiation. In addition, the Student's t test revealed that the T₂* value could determine grouped T stages (T0 vs. T1–3: 17.0 ± 2.9 ms vs. 25.2 ± 6.2 ms; T0–1 vs. T2–3: 17.3 ± 3.0 ms vs. 27.1 ± 5.3 ms; and T0–2 vs. T3: 18.8 ± 4.2 ms vs. 27.8 ± 5.6 ms, all p(s) < 0.001). ROC analysis indicated that the T₂* value could detect ESCCs (cutoff, 20 ms), and discriminate between stages T0–1 and T2–3 (cutoff, 21.3 ms) and between T0–2 and T3 (cutoff, 20.4 ms). CONCLUSION: The T₂* value can be an additional quantitative indicator for detecting ESCC except for stage T1 cancer, and can preoperatively discriminate between some T stages and between anatomic stages of this tumor.

Expression of CD163 in children with Epstein-Barr virus infection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical significance of CD163 in the diagnosis and the evaluation of severity and prognosis of childhood hemophagocytic lymphohistiocytosis (HLH).</p><p><b>METHODS</b>Ninety-four children were classified into Epstein-Barr virus (EBV)-positive (n=55) and EBV-negative groups (n=39; control group). The EBV-positive group was subgrouped into infectious mononucleosis (IM; n=47) and HLH (n=8). Serum levels of soluble CD163 were measured using ELISA. Expression of CD163 on mononuclear cells was detected by flow cytometry.</p><p><b>RESULTS</b>The serum levels of soluble CD163 were>10 000 ng/mL in all eight HLH patients (>30 000 ng/mL in 3 cases). The mean serum levels of soluble CD163 in the HLH group were significantly higher than in the control and IM groups (P<0.05). The serum levels of soluble CD163 in EBV-positive children were positively correlated with EBV-DNA copies and serum levels of ferritin and LDH, but were negatively correlated with white blood cell count, neutrophil count, hemoglobin and platelet count. The follow-up after treatment for three HLH patients showed that serum levels of soluble CD163 were significantly reduced, but the soluble CD163 levels rebounded in one patient who was complicated by fungal pneumonia infection.</p><p><b>CONCLUSIONS</b>The levels of serum soluble CD163 may be related to the severity in children with HLH. The EBV-positive children with soluble CD163 levels >10 000 ng/mL should be considered the possibility of HLH.</p>

Association of tag SNPs of GPx-3 with essential hypertension in rural Han Chinese in Fuxin, Liaoning, China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Genetic mechanisms contribute to blood pressure regulation. This study investigated whether glutathione peroxidase (GPx-3) tag single nucleotide polymorphisms (SNPs) are associated with hypertension in the rural areas of Fuxin county, Liaoning province, China.</p><p><b>METHODS</b>Indigenous Fuxin Han people participated, 523 unrelated hypertensives and 547 controls were recruited. All tag SNPs of GPx-3 gene were selected. We estimated SNP allele frequency in DNA pools with pyrosequencing.</p><p><b>RESULTS</b>Before Bonferroni correction, C allele frequency for rs8177417 was significantly higher in hypertensives than those in controls (23.4% vs. 19.3%, P = 0.014); T allele frequency for rs3828599 was significantly lower in hypertensives than those in controls (35.6% vs. 40.8%, P = 0.009). However, when a Bonferroni correction for multiple testing was applied, only the polymorphisms rs3828599 of GPx-3 gene was associated with hypertension (P = 0.045, OR: 0.833, 95%CI: 0.695 - 0.998).</p><p><b>CONCLUSION</b>The polymorphism of rs3828599 of GPx-3 gene might be associated with hypertension in rural Han Chinese from Fuxin, Liaoning.</p>

Preparation and bioactivity evaluation of streptavidin-tagged human interleukin-15 fusion protein / 南方医科大学学报

<p><b>OBJECTIVE</b>To obtain streptavidin-tagged human interleukin-15 (SA/hIL15) fusion protein and evaluate its bioactivity.</p><p><b>METHODS</b>pET24a-6His-SA-hIL-15 and pET32a-hIL-15-SA-6His plasmids were constructed and expressed in BL 21(DE3) host bacteria to generate the fusion protein. The recombinant fusion protein IL-15/SA was purified using Ni-NTA affinity chromatography and refolded, and the efficiency of surface modification of the fusion protein on biotinylated cells was examined by fluorescence-activated cell sorting. CCK-8 method was used to evaluate the effect of IL-15/SA fusion protein in inducing the proliferation of human peripheral-blood lymphocyte (PBL) cells stimulated by PHA.</p><p><b>RESULTS</b>The recombinant SA-hIL-15 and hIL15-SA fusion proteins were highly expressed in BL21(DE3) at about 20% of the total bacterial proteins. The purified hIL15-SA fusion protein exhibited a bifunctionality by promoting the proliferation of PBL cells activated by PHA and high-affinity binding to biotinylated cell surface mediated by SA, with a cell surface modification efficiency exceeding 95%. SA-hIL-15 showed a 4-fold higher hIL15 bioactivity than hIL15-SA.</p><p><b>CONCLUSION</b>SA/hIL-15 bifunctional fusion protein has been successfully obtained to facilitate the future development of hIL-15-surface-modified cancer cell vaccine.</p>

Surgical management of tumor in the conjunctive area among neck, thorax and axilla / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the surgical management of tumors in the conjunctive area among the neck, thorax and axilla and its efficacy.</p><p><b>METHODS</b>From Oct. 1999 to March 2006, eleven cases with benign tumors in the area between the neck, thorax and axilla were collected and analysed. Among them, five neurilemmoma, three neurofibroma, two chondroma and one meningioma, respectively. CT scans showed that the neck, thorax and axilla were affected simultaneously more or less. In this group, direct invasion to the vertebra was found in 4 cases, and spinal compression in one patient. After the disarticulation and displacement of the clavicle, the tumor was removed with the structures of importance in the neck, thorax and axilla under direct views. A bypass was constructed between the axillary vein and the medial end of the subclavian vein.</p><p><b>RESULTS</b>All these tumors were completely removed, injury of vertebral artery was encounter in two cases, and immediate repair was successfully carried out. No other serious complications were found in this group. During the follow-up period from 3 months to 3 years and 5 months, local recurrence was found in no patient.</p><p><b>CONCLUSIONS</b>Benign tumors in the area between neck, thorax and axilla could be successfully dissected and removed with displacement of the medial portion of the clavicle.</p>

Determining the relationship between coronary artery and coronary sinus and its tributaries using multislice computed tomography / 中华心血管病杂志

<p><b>OBJECTIVE</b>To obtain the coronary artery and coronary sinus (CS) and its tributaries imaging with multislice computed tomography (MSCT), measure the distance between coronary artery and CS and its tributaries and analyze their spatial relationships.</p><p><b>METHODS</b>The MSCT scans of 117 patients (67 men, 50 women, age 56 +/- 10 years) were obtained, 3D image reconstructed and the vessels courses evaluated. The concomitant distances and spatial relationships of the vessels were determined.</p><p><b>RESULTS</b>Right coronary artery domination was found in 107 cases (91.4%), left coronary artery domination in 7 cases (6.0%), and co-domination in 3 cases (2.6%). Left circumflex artery (LCX) was concomitant with CS or the great cardiac vein (GCV) in 81 cases (69.2%), intersected with left posterior vein in 62 cases (53.0%) and with middle cardiac vein (MCV) in 5 cases (4.3%), respectively. The dominant coronary artery branched out into the posterior descending artery (PDA) and the left posterior artery (LPA) in 112 cases (95.7%). PDA was concomitant with MCV in 93 cases (79.5%) and intersected with MCV in 44 cases (37.6%). LPA was intersected with MCV in 106 cases (90.6%), and concomitant with CS in 50 cases (42.7%).</p><p><b>CONCLUSIONS</b>MSCT is a reliable tool to visualize the relationship between coronary artery and CS and its tributaries. Owing to the multiple possibilities inherent to this technique, MSCT has broad potential for more clinical use.</p>

Effect of Fuzheng Jiangan formula on liver fibrosis induced by albumin in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of Fuzheng Jiangan formula( FZJGF) on liver fibrosis using immune induced liver fibrosis rat model.</p><p><b>METHOD</b>The rat models with immunity liver fibrosis were induced by the human serum albumin. Rats were treated with normal saline, FZJGF (9. 85,39. 4 g x kg(-1) , two dosage groups) and Colchicine (0. 000 1 g . kg(-1) ). The activities of ALT, AST, contents of ALB and TP, and A/G, The contents of Laminin (LN), Hyaluronic acid (HA) and collagen type IV (IV-C) in rat serum were measured by radioimmunoassay method. The level of hydroxyproline (Hyp) in liver was detected by chemistry method. The pathological changes of liver tissue were observed by HE and Von-Gieson staining.</p><p><b>RESULT</b>FZJGF could significant decrease the serum activities of ALT and AST, and increase the levels of TP,Alb and ratio A/G. The levels of LN, HA and IV-C were decreased significantly after the treatment using FZJGF. The pathological improvements were observed. FZJGF could markedly alleviate the deposition of collageneous fiber, and reduce the liver pseudoluboli and the fibrosis scores in the liver tissue compared with model group.</p><p><b>CONCLUSION</b>FZJGF can inhibit formation and development of rat hepatic fibrosis induced by the human serum albumin.</p>