Integrated analysis of whole genome and transcriptome sequencing reveals a frameshift mutation associated with recessive embryonic lethality in Holstein cattle.

Embryo loss is an important factor affecting fertility in dairy production. HH2 was identified as a haplotype on chromosome 1 associated with embryonic lethality in Holstein cattle. In the current study, both short- and long-read WGS was performed on four carriers and four non-carriers of HH2 to screen for variants in concordance with HH2 haplotype status. Sequence variation analysis revealed five putative functional variants of protein-coding genes, including a frameshift mutation (g.107172616delT) in intraflagellar transport protein 80 (IFT80) gene. Transcriptome analysis of whole blood indicated that no gene exhibited significantly differential expression or allele-specific expression between carriers and non-carriers in the candidate region. This evidence points to g.107172616delT as the highest priority causative mutation for HH2. Protein prediction reveals that the frameshift mutation results in a premature stop codon to reduce the peptide chain from 760 to 383 amino acids and greatly alters the structure and function of IFT80 protein. Our results demonstrate that the use of a combination of multiple high-throughput sequencing technologies is an efficient strategy to screen for the candidate causative mutations responsible for Mendelian traits, including genetic disorders.

Caspofungin combined with TMP/SMZ as a first-line therapy for moderate-to-severe PCP in patients with human immunodeficiency virus infection.

OBJECTIVES: The effectiveness of trimethoprim/sulfamethoxazole (TMP/SMZ) for pneumocystis pneumonia (PCP) is limited with adverse events. Caspofungin, by inhibiting the cyst form of Pneumocystis jirovecii, may be an alternative therapy for PCP. However, the availability of clinical data about caspofungin combined with TMP/SMZ in the treatment of PCP in HIV-infected patients is limited. Thus, we aimed to examine the clinical effectiveness and safety of caspofungin combined with TMP/SMZ as a first-line therapy for moderate-to-severe PCP in HIV-infected patients. METHODS: From January 2017 to December 2019, data of HIV-infected patients with moderate-to-severe PCP who received either TMP/SMZ alone or caspofungin combined with TMP/SMZ as first-line therapy were retrospectively reviewed to assess the effectiveness and safety of each regimen. The Kaplan-Meier curve and log-rank test were used for survival analysis. RESULTS: A total of 278 patients met the criteria. The overall positive response rate of PCP treatment was 48.92%, and the overall all-cause in-hospital mortality rate was 33.09%. Patients who received combination therapy consisting of caspofungin and TMP/SMZ had a better positive response rate (59.44% vs. 37.78%, P < 0.001) and lower all-cause in-hospital mortality rate (24.48% vs. 42.22%, P = 0.003). Also, patients who received combination therapy had higher survival rate during a hospital stay (75.52% vs. 57.78%, P = 0.004), and those who received longer combination therapy were more likely to have higher survival rate (P = 0.042). We found that age (P = 0.019), CD4 cell count (P = 0.001) and therapeutic regimen (P = 0.002) were significant risk factors for all-cause in-hospital mortality rate in univariate analysis. In multivariate analysis, only CD4 cell count and therapeutic regimen were statistically significant factors associated with all-cause in-hospital mortality rate. Patients with a CD4 count of > 30 cells/µL and patients who received combination therapy consisting of caspofungin and TMP/SMZ were more likely to survive from PCP (P = 0.011 and P = 0.002, respectively). There were no additional severe adverse events caused by adding caspofungin. CONCLUSIONS: For HIV-infected patients with moderate-to-severe PCP, combination therapy with caspofungin and TMP/SMZ is an effective and promising first-line therapy with no greater number of adverse events compared with TMP/SMZ monotherapy. Patients who received caspofungin had better positive response rates and lower all-cause in-hospital mortality rates. Also, we recommend early initiation of caspofungin.

[Efficacy and safety of early initiation of sacubitril-valsartan therapy in patients with acute decompensated heart failure].

Objective: To assess the efficacy and safety of the initiation of sacubitril-valsartan （ARNI） therapy, as compared with ACEI therapy, after hemodynamic stabilization among patients hospitalized for acute decompensated heart failure （ADHF）. Methods: A total of 199 hospitalized patients for ADHF in our department from January 2017 to June 2019 were included in this retrospective analysis. According to the medication early after hemodynamic stabilization, patients were divided into ARNI group (n=92) and ACEI group (n=107). Among the included patients, 61 patients with newly diagnosed heart failure at the time of admission were also divided into ARNI group (n=30) and ACEI group (n=31) according to the applied medication. Clinical baseline data and follow-up results of enrolled patients were collected through the electronic medical records at admission, outpatient and telephone follow-up. The primary effectiveness observation index was left ventricular ejection fraction (LVEF) and left ventricular end diastolic dimension (LVEDD) measured by echocardiography; the secondary observation index was death from any causes and hospitalization for heart failure. Safety outcomes were the incidences of symptomatic hypotension, worsening renal function, hyperkalemia, and angioedema. Results: The clinical baseline characteristics were similar between ARNI group and ACEI group（all P>0.05）. The duration of follow up was (15.2±6.5) months in all patients enrolled, (12.3±5.0) months in ARNI group, and (18.2±6.5) months in ACEI group. At the end of follow-up, prevalence of an absolute LVEF increase of more than 5% was 48.9% (45/92) in ANRI group and 25.2% (27/107) in ACEI group (P=0.001). Percent of LVEF increase to more than 50% was 17.4% (16/92) in ANRI group and 3.7% (4/107) in ACEI group (P=0.001). Percent of patients with more than 10 mm LVEDD reduction was 14.1% (13/92) in ANRI group and 3.7% (4/107) in ACEI group (P=0.009). All-cause mortality rate was 5.7% (5/88) in ARNI group and 15.3% (13/85) in ACEI group (P=0.038). Rate of re-hospitalization due to heart failure was 50% (46/92) in ARNI group and 71% (76/107) in ACEI group（P=0.002）.The rates of symptomatic hypotension, worsening renal function, hyperkalemia, and angioedema were similar between ARNI group and ACEI group (all P>0.05). In patients with first diagnosed heart failure，percent of LVEF increase to more than 50% was 30% (9/30) in ANRI group and 6.5% (2/31) in ACEI group (P=0.017). Percent of more than 10 mm LVEDD reduction was 26.7%(8/30) in ANRI group and 3.2%(1/31) in ACEI group (P=0.012). Percent of an absolute LVEF increase of more than 5% was 53.3% (16/30) in ANRI group and 51.6% (16/31) in ACEI group (P=0.893). Re-hospitalization due to heart failure was 23.3% (7/30） in ARNI group and 73.3% (11/31) in ACEI group（P<0.01）. Rate of all-cause death tended to be lower in patients receiving ARNI (3.4% (1/29)) as compared to patients receiving ACEI (13.0% (3/23), P=0.197）. Conclusions: Among patients with heart failure with reduced ejection fraction hospitalized for ADHF, the initiation of ARNI therapy after hemodynamic stabilization is associated with a more significant improvement of cardiac remodeling and pump function than ACEI therapy and satisfactory safety. In ADHF patients with first diagnosed heart failure, initiation of ARNI therapy after hemodynamic stabilization can more effectively improve cardiac remodeling and pump function than treatment with ACEI.

[A single-center retrospective study on axillary evaluation in 1 557 breast ductal carcinoma in situ patients between 2006 and 2016].

Objectives: To examine the influence factors on axillary evaluation in ductal carcinoma in situ (DCIS) patients, and the prognosis of different choices of axillary evaluation in a single-center retrospective study. Methods: Totally 1 557 DCIS patients admitted in Department of Breast Surgery, Fudan University Shanghai Cancer Center from January 2006 to November 2016 were retrospectively enrolled. All patients were female. The median age was 49 years (range: 21 to 85 years). Surgical methods included modified radical mastectomy, simple mastectomy (with or without axillary evaluation) and breast conservation surgery (with or without axillary evaluation). Axillary evaluation included axillary lymph node dissection (ALND) and sentinel lymph node biopsy (SLNB). T tests, χ(2) test and Logistic regression analysis was used to analyze influence factors on axillary evaluation, respectively. Kaplan-Meier curve and Log-rank analysis were used to evaluate recurrence-free survival (RFS) and loco-regional recurrence-free survival (LRRFS) in patients with different surgical methods. Results: Among the 1 557 DCIS patients, there were 1 226 cases received axillary evaluation, while 331 cases not received axillary evaluation. Patients were separated into 3 groups by different axillary evaluation choices: SLNB group (957 cases, 61.46%), ALND group (197 cases, 12.65%) and no evaluation group (403 cases, 25.88%). The patients in SLNB group increased significantly (P=0.000), from 3.85% (60/1 557) in 2006 to 75.19% (1 170/1 557) in 2016. The independent influence factors of receiving axillary evaluation were high nuclear grade (OR=3.191, 95%CI: 1.722 to 5.912, P=0.001) and tumor size>15 mm (OR=1.698, 95%CI: 1.120 to 2.573, P=0.012). Also, patients received breast conservation surgery were more likely to refuse axillary evaluation (OR=0.155, 95%CI: 0.103 to 0.233, P=0.000). There were no significant differences in RFS and LRRFS in patients with different axillary evaluation choices. Conclusions: The investigation in trends and influence factors of different axillary evaluation choices provided basis on surgical precision medicine in DCIS patients. Patients received SLNB increased significantly. The independent influence factors of axillary evaluation were nuclear grade, tumor size and surgical methods. There was no significant differences in prognosis among the groups receiving different axillary evaluations.

[An analysis of the new "three tubes" method in the treatment of spontaneous esophageal rupture].

To investigate the clinical efficacy, feasibility and safety of new "three tubes" method in the treatment of spontaneous esophageal rupture. A total of 22 patients with spontaneous esophageal rupture were retrospectively analyzed. Through the new "three tubes" method of treatment, patients achieved leak cured with reduced hospital stay, less medical expenses and early resumption of oral diet. The new "three tubes" method for spontaneous esophageal rupture has the advantages of easy handling, minimal invasion, few complication and exact curative effect.

[Magnetic resonance imaging features of tear outflow and structural changes of the lacrimal sac and nasolacrimal duct in normal subjects with eyes closed].

Objective: To investigate the static and dynamic features of the normal lacrimal sac (LS)-nasolacrimal duct (NLD), and find out if tear flows while the subject is in supine position with eyes closed. Methods: Cross-sectional study. Healthy volunteers and patients whose head were examined by MRI in Guangdong Provincial Hospital of Integrated Traditional Chinese and Western Medicine with normal LS-NLD were recruited. The normal lacrimal ducts of the candidates were scanned, while candidates remained in supine position, by static or dynamic imaging procedures with a 1.5T MRI system, which adopted the thinnest slice thickness, consecutive sections (no spacing) and two scanning planes (axial and coronal planes of LS-NLD). The static and dynamic imaging procedures were done in June and July 2014 and October 2015, respectively. The static imaging was performed with conventional T(2)WI, contrast-enhanced 3D FIESTA-C, contrast-enhanced T(1)WI and heavy T(2)WI pulse sequences under normal physiological condition, and 104 eyes with normal LS-NLD from 39 patients and 13 healthy volunteers (21 male and 31 female) aged 10 to 75 years were scanned with the static procedure. The dynamic imaging was done with heavy T(2)WI pulse sequence. The coronal dynamic imaging was done only after 0.9% NaCl was instilled in the conjunctival sacs, and 20 eyes with normal LS-NLD of 10 volunteers (6 male and 4 female) aged 23 to 53 years were scanned with the coronal dynamic procedure. The axial dynamic imaging was done under normal physiological condition and after 0.9% NaCl was instilled in the conjunctival sacs respectively, and 20 eyes with normal LS-NLD of 10 volunteers (8 male and 2 female) aged 20 to 40 years were scanned with the axial dynamic procedure. The incidences on the axial dynamic imaging under the two conditions were compared with χ(2) test. Results: A total of 104 eyes with normal LS-NLD (52 candidates) were scanned with the static imaging sequences. In all LS-NLDs, an indentation located at the posterior-lateral side wall of the junction of LS-NLD was discovered, and its depth was variable. In 28.8% (30/104) of LS, 64.4% (67/104) of the junctions and 22.1% (23/104) of NLD, lumens were closed. As for the coronal dynamic imaging (20 eyes with normal LS-NLD in 10 subjects), segmental tear transit was observed in 15/20 of NLD. As for the axial dynamic imaging (20 eyes with normal LS-NLD in 10 subjects), after 0.9% NaCl was instilled, dynamic changing of their lumen size was revealed in 17/20 of LS-NL and the occurrence rate of the lumen size changing (29.3%, 94/321, the number of the axial slices with lumen changes/total number of the axial slices imaged) was higher than that (16.4%,52/317) under normal physiological condition. The difference between the two rates was statistically significant (χ(2)=14.993, P<0.001). LS contained more tear than NLD. Conclusion: While the candidates are in supine position with eyes closed (absolutely no blinking), an indentation is at the posterior-lateral side wall of the junction of LS-NLD, and there are autonomous, segmental lumen open-close actions in normal LS-NLD and tear transit in their lumens. (Chin J Ophthalmol, 2018, 54: 205-211).

Characterization of the alkaline/neutral invertase gene in Dendrobium officinale and its relationship with polysaccharide accumulation.

Dendrobium officinale is one of the most well-known traditional Chinese medicines, and polysaccharide is its main active ingredient. Many studies have investigated the synthesis and accumulation mechanisms of polysaccharide, but until recently, little was known about the molecular mechanism of how polysaccharide is synthesized because no related genes have been cloned. In this study, we cloned an alkaline/neutral invertase gene from D. officinale (DoNI) by the rapid amplification of cDNA ends (RACE) method. DoNI was 2231 bp long and contained an open reading frame that predicted a 62.8-kDa polypeptide with 554-amino acid residues. An alkaline/neutral invertase conserved domain was predicted from this deduced amino acid sequence, and DoNI had a similar deduced amino acid sequence to Setaria italica and Oryza brachyantha. We also found that DoNI expression in different tissues was closely related to DoNI activity, and more importantly, polysaccharide level. Our results indicate that DoNI is associated with polysaccharide accumulation in D. officinale.

Relationship between TBX20 gene polymorphism and congenital heart disease.

Congenital heart disease in children is a type of birth defect. Previous studies have suggested that the transcription factor, TBX20, is involved in the occurrence and development of congenital heart disease in children; however, the specific regulatory mechanisms are yet to be evaluated. Hence, this study aimed to evaluate the relationship between the TBX20 polymorphism and the occurrence and development of congenital heart disease. The TBX20 gene sequence was obtained from the NCBI database and the polymorphic locus candidate was predicted. Thereafter, the specific gene primers were designed for the restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) of DNA extracted from the blood of 80 patients with congenital heart disease and 80 controls. The results of the PCR were subjected to correlation analysis to identify the differences between the amplicons and to determine the relationship between the TBX20 gene polymorphism and congenital heart disease. One of the single nucleotide polymorphic locus was found to be rs3999950: c.774T>C (Ala265Ala). The TC genotype frequency in the patients was higher than that in the controls, similar to that for the C locus. The odds ratio of the TC genotypes was above 1, indicating that the presence of the TC genotype increases the incidence of congenital heart diseases. Thus, rs3999950 may be associated with congenital heart disease, and TBX20 may predispose children to the defect.

[Assessment and significance of human epidermal growth factor receptor 2 gene status in breast cancer with polysomy of chromosome 17].

OBJECTIVE: To explore the impact of polysomy of chromosome 17 on the interpretation of human epidermal growth factor receptor 2 (HER2) status and its clinical significance in breast cancers. METHODS: Clincopathological data of 338 breast cancer patients were collected, and the association of clinicopathological characteristics with polysomy of chromosome 17 was analyzed. Based on the American Society of Clinical Oncology /College of American Pathologists updated recommendations of HER2 testing in breast cancer in 2013, the relationships between the copy number of centromeres on chromosome 17 (CEP17) and HER2 status, other important biological markers and clincopathological features of breast cancer were analyzed statistically. RESULTS: Immunohistochemical examination revealed that among the 338 breast cancer cases, there were 9 (2.7%) HER2-negative cases, 309 (91.4%) HER2-equivocal cases, and 20 (5.9%) HER2-positive cases. The FISH analysis showed that there were 226 (66.9%) HER2-negative cases, 13 (3.8%) HER2-equivocal cases, and 99 (29.3%) HER2-positive cases. Among the 338 breast cancer patients, 42 (12.4%) cases were identified as chromosome 17 polysomy (polysomy 17), 291 (86.1%) cases were chromosome 17 disomy, and 5 (1.3%) cases were chromosome 17 monosomy. Polysomy 17 showed significant correlation with the intensity of HER2 protein expression (P=0.046). The HER2 FISH results and HER2 gene copy number of polysomy 17 cases showed significant differences from the non-polysomy 17 cases (P<0.001). There were no significant differences of HER2/CEP17 ratio between the polysomy 17 and non-polysomy 17 groups (P=0.930). Polysomy 17 cases showed a significant relationship with tumor grading and Ki-67 index (P<0.05), but not with the age of patient, tumor size, lymph node metastasis, ER and PR expressions, and effect evaluation of neoadjuvant chemotherapy (P>0.05). CONCLUSIONS: In the breast cancer with polysomy of chromosome 17, assessment of the absolute copy number of HER2 signals is even more important, and the HER2/CEP17 ratio and HER2 protein expression levels should be comprehensively evaluated together.

[Clinical significance of expression of PI3Kp110α in breast cancer].

Objective: To study the clinical significance of PI3Kp110α expression in breast cancer. Methods: The expressions of PI3Kp110α, HER2, PTEN, p-Akt and Ki-67 in invasive ductal carcinoma (IDC), adjacent ductal carcinoma in situ (DCIS) and normal breast tissue were detected by immunohistochemistry in 102 cases of breast cancer. The expression of PI3Kp110α in IDC was compared with those in DCIS and normal breast tissues. Correlations between expression of PI3Kp110α and expression of HER2, PTEN, p-Akt and Ki-67 index in IDC were analyzed. Correlation between expression of PI3Kp110α and stage of IDC was studied as well. Results: In the normal breast tissues, there were 97 cases (95.1%) with low level and 5 cases (4.9%) with high level expression of PI3Kp110α. In the DCIS tissues, there were 67 cases (65.7%) with low level and 35 cases (34.3%) with high level expression of PI3Kp110α. In the IDC tissues, there were 14 cases (13.7%) with low level and 88 cases (86.3%) with high level expression of PI3Kp110α. The difference between expression of PI3Kp110α in normal breast tissue, DCIS and IDC was significant (P<0.001). In the IDC tissues, expression of PI3Kp110α was negatively correlated with expression of HER2 (rs=-0.213, P=0.032) and PTEN (rs=-0.197, P=0.047), while was not significantly correlated with expression of p-Akt (P=0.119) and Ki-67 index (P=0.636). In contrast, expressions of HER-2 and p-Akt were positively correlated with Ki-67 index (P=0.001, P=0.035), while expression of HER2 was not correlated with p-Akt (P=0.177). Expression of PI3Kp110α was negatively correlated with T stage and TNM stage (P=0.003, P=0.016), while not correlated with N stage and M stage(both P>0.05). Expression of HER-2 was positively correlated with T stage (P=0.037), while not correlated with N stage, M stage and TNM stage (P>0.05 for all). Neither Ki-67 index nor expression of PTEN and p-Akt (P=0.194) were correlated with T stage, N stage, M stage and TNM stage. Conclusions: Expression of PI3Kp110α plays an important role in oncogenesis and development of breast cancer. Based on the fact that expression of PI3Kp110α is negatively correlated with expression of HER2 and PTEN and with T stage and TNM stage, we may conclude that increased expression of PI3Kp110α is another independent pathway in breast cancer development, and breast cancer patients with high expression of PI3Kp110α may have a better prognosis.

[Characteristic and clinical significance of DNA methyltransferase 3B overexpression in endometrial carcinoma].

OBJECTIVE: To determine the clinicopathological significance of the DNA methyltransferase 3B (DNMT3B) overexpression in endometrial carcinomas and to evaluate its correlation with hormone receptor status. METHODS: Immunohistochemistry was performed to assess the expression of DNMT3B and hormone receptors in 104 endometrial carcinomas. RESULTS: DNMT3B overexpression occurred frequently in endometrioid carcinoma (EC, 54.8%) more than in nonendometrioid carcinoma (NEC, 30.0%) with statistical significance (P=0.028). Furthermore, there was a trend that EC with worse clinico-pathological variables and shorter survival had a higher DNMT3B expression, and the correlation between DNMT3B and tumor grade reached statistical significance (P=0.019).A negative correlation between DNMT3B and estrogen receptor (ER) or progesterone receptor (PR) expression was found in EC. NMT3B overexpression occurred frequently in the ER or PR negative subgroups (78.9%, 86.7%) more than in the positive subgroups (47.7%, 47.8%) with statistical significance (P=0.016, P=0.006). In addition, the DNMT3B overexpression increased in tumors with both ER and PR negative expression (92.9%, P=0.002). However, no such correlation was found in NEC (P>0.05). Sequence analyses demonstrated multiple ER and PR binding sites in the promoter regions of DNMT3B gene. CONCLUSION: This study showed that the expression of DNMT3B in EC and NEC was different. DNMT3B overexpression in EC was associated with the worse clinicopathological variables and might have predictive value. The methylation status of EC and NEC maybe different. In addition, in EC, DNMT3B overexpression negatively correlated with ER or PR expression. In NEC, the correlation between DNMT3B and ER or PR status was not present.

Long Noncoding RNA: its partners and their roles in cancer.

RNA transcripts, which do not encode proteins, have received considerable attention in recent years. These non-coding RNAs are classified into two groups: small non-coding RNAs and long non-coding RNAs (lncRNAs). Increasing evidence suggests that lncRNAs are emerging as key regulators in many biological processes. However, knowledge of the underlying mechanisms whereby they act is still limited. Here, we try to elucidate the way that lncRNAs function in the context of DNA, RNA and protein interaction networks. It is noteworthy that lncRNA and another type of non-coding RNA microRNA (miRNA) may 'talk' to each other more frequently than ever expected. Additionally, lncRNAs display aberrant expression profiles in different kinds of cancers, with their potential roles in carcinogenesis and cancer metastasis. We summarize the effect of some cancer related lncRNAs upon tumor biological events, including cell proliferation, apoptosis, invasion and metastasis. Finally, we focus on the clinical value of lncRNAs, considering their potential application in cancer diagnosis, prognosis and therapeutic intervention.

Correlation of Left Ventricular Diastolic Function and Left Ventricular Geometry in Patients with Obstructive Sleep Apnoea Syndrome.

BACKGROUND: The aim of this study is to evaluate the correlation of the left ventricular diastolic function and the left ventricular geometry in patients with obstructive sleep apnoea syndrome (OSAS) by echocardiography. METHODS: The 181 patients diagnosed with OSAS were divided into the normal geometry group (NG), the concentric remodelling group (CR), the eccentric hypertrophy group (EH) and the concentric hypertrophy group (CH). Pearson correlation analysis and multiple linear regression analysis were performed toward the correlation of the left ventricular diastolic function and the left ventricular geometry. RESULTS: The E peak in the EH and CH group was significantly reduced, with significant difference; the E/A, Em, Am and Em/Am was reduced in the order of the CR, EH and CH groups, while E/Em was increased, and the difference was significant. Pearson correlation analysis revealed that the Em/Am showed significant negative correlations with the left ventricular mass index (LVMI) [r = -0.419] and relative wall thickness (RWT) [r = -0.289], while the E/Em was significantly positively correlated with the LVMI (r = 0.638) and RWT [r = 0.328] (p < 0.001). Multiple linear regression analysis revealed that LVMI and RWT had influence on the Em/Am and E/Em (r2 = 0.402, r2 = 0.107, p < 0.001). The left ventricular diastolic dysfunction was the worst in the CH group. CONCLUSIONS: There was correlation between the left ventricular diastolic dysfunction and the changes in cardiac geometry.

Regulatory effect of iron regulatory protein-2 on iron metabolism in lung cancer.

Iron metabolism plays an important role in the pathogenesis of lung cancer. This study aimed to investigate the effect of gene silencing of iron regulatory protein-2 (IRP2) on mRNA and protein expression of transferrin (Tf), transferrin receptor (TfR), and ferritin (Fn) in A549 lung cancer cells. A549 cells were cultured and divided into a liposome control group, a liposome + oligonucleotide (SCODN) control group, and a Lipofectamine + antisense oligonucleotide (ASODN) group. RT-PCR and Western blotting were used to detect mRNA and protein expression of Tf, TfR, and Fn. We found no significant change in Tf mRNA expression among the 3 groups (P = 0.078). TfR and Fn mRNA expressions in the ASODN group notably decreased compared to the liposome and SCODN groups (P < 0.01). IRP2 and TfR protein expressions in the ASODN group were significantly lower than in the liposome or SCODN groups (P < 0.05), whereas no significant change in Tf protein expression was observed between the 3 groups (P = 0.088). Fn protein expression in the ASODN group was significantly higher than in the liposome or SCODN group (P < 0.05). IRP2 can regulate the expression of TfR and Fn by changing its own protein expression and thereby regulate iron metabolism.

Ambient levels and temporal variations of PM2.5 and PM10 at a residential site in the mega-city, Nanjing, in the western Yangtze River Delta, China.

The deteriorating air quality in eastern China including the Yangtze River Delta is attracting growing public concern. In this study, we measured the ambient PM10 and fine PM2.5 in the mega-city, Nanjing at four different times. The 24-h average PM2.5 and PM10 mass concentrations were 0.033-0.234 and 0.042-0.328 mg/m(3), respectively. The daily PM10 and PM2.5 concentrations were 2.9 (2.7-3.2, at 95% confidence interval) and 4.2 (3.8-4.6) times the WHO air quality guidelines of 0.025 mg/m(3) for PM2.5 and 0.050 mg/m(3) for PM10, respectively, which indicated serious air pollution in the city. There was no obvious weekend effect. The highest PM10 pollution occurred in the wintertime, with higher PM2.5 loadings in the winter and summer. PM2.5 was correlated significantly with PM10 and the average mass fraction of PM2.5 in PM10 was about 72.5%. This fraction varied during different sampling periods, with the lowest PM2.5 fraction in the spring but minor differences among the other three seasons.

[Tanshinone â ¡A activates PI3K/AKT signaling pathway to inhibit the apoptosis of mice cochlear pericytes induced by high glucose].

Objective: To investigate whether tanshinone ⅡA can protect the apoptosis of mice cochlear pericytes induced by high glucose and its specific protective mechanism, so as to provide experimental evidence for the prevention and treatment of diabetic hearing loss. Methods: C57BL/6J male mice were used to prepare type 2 diabetes model, which were divided into normal (NG) group, diabetic (DM) group, diabetic+tanshinone ⅡA (HG+tanshinone ⅡA) group and tanshinone ⅡA group. Each group had 10 animals. Primary cochlear pericytes were divided into NG group, HG group (high glucose 35 mmol/L), HG+tanshinone ⅡA (1, 3, 5 µmol/L) group, HG+Tanshinone ⅡA+LY294002 (PI3K/AKT pathway inhibitor) group, LY294002 group, tanshinone ⅡA group and DMSO group. Auditory brainstem response (ABR) was used to measure hearing threshold. Evans blue was used to detect the permeability of blood labyrinth barrier in each group. TBA methods were used to detect oxidative stress levels in various organs of mice. Morphological changes of stria vascularis were observed by hematoxylin-eosin staining (HE). Evans blue was used to detect the vascular labyrinth barrier permeability in cochlea. The expression of apoptosis protein in stria vascularis pericytes was observed by immunofluorescence. Pericytes apoptosis rate was observed by flow cytometry. DCFH-DA was combined with flow cytometry to detect intracellular ROS content, and Western blot was used to detect the expression of apoptotic proteins (Cleaved-caspase3, Bax), anti-apoptotic proteins (BCL-2) and pathway proteins (PI3K, p-PI3K, AKT, p-AKT). SPSS software was used for statistical analysis. Independent sample t test was performed, and P<0.05 was considered statistically significant. Results: Animal experiments: Tanshinone ⅡA decreased the hearing threshold of DM group [(35.0±3.5) dB SPL vs. (55.3±8.1) dB SPL] (t=4.899, P<0.01), decreased the oxidative stress level in cochlea (t=4.384, P<0.05), improved the structure disorder, atrophy of cochlea vascular lines, vacuole increased phenomenon. Tanshinone ⅡA alleviated the increased permeability of the blood labyrinth barrier [Evans blue leakage (6.84±0.27) AU vs. (8.59±0.85) AU] in the cochlea of DM mice (t=2.770, P<0.05), reversed the apoptotic protein: Caspase3 (t=4.956, P<0.01) and Bax (t=4.388, P<0.05) in cochlear vascularis. Cell experiments: Tanshinone ⅡA decreased intracellular ROS content in a concentration-dependent way (t=3.569, P<0.05; t=4.772, P<0.01; t=7.494, P<0.01); Tanshinone ⅡA decreased apoptosis rate and apoptotic protein, and increased the expression of anti-apoptotic protein, p-PI3K/PI3K and p-AKT/AKT in concentration-dependent manner (all P values<0.05); LY294002 reversed the protective effect of tanshinone ⅡA on pericytes apoptosis (all P values<0.05). Conclusion: Tanshinone ⅡA can inhibit the apoptosis of cochlear pericytes induced by high glucose by reducing oxidative stress level and activating PI3K/AKT signaling pathway under high glucose environment, thus playing a protective role in diabetic hearing loss.

Association of Computed Tomography Radiomics Signature with Progression-free Survival in Neuroblastoma Patients.

AIMS: To investigate the association of computed tomography radiomics signature with progression-free survival (PFS) in neuroblastoma patients. MATERIALS AND METHODS: We retrospectively included 167 neuroblastoma patients who were divided into a training set and a test set through stratified sampling at a ratio of 7:3. Regions of interest of the primary tumours were delineated on pretreatment contrast-enhanced computed tomography images and radiomics features were extracted from them. The intraclass correlation coefficient, Pearson correlation coefficient, and least absolute shrinkage and selection operator Cox regression algorithm were applied to select radiomics features and construct the radiomics signature. The effectiveness of the signature in predicting PFS was evaluated using the concordance index (C-index) and 95% confidence interval in both the training and the test sets. The time-dependent receiver operator characteristic curve of the radiomics signature was plotted and the area under the curve (AUC) was calculated. A calibration curve was used to assess the difference between the predicted probability of the radiomics signature and the observed probability at different time points. RESULTS: The radiomics signature was composed of six features, which achieved a C-index of 0.733 (95% confidence interval 0.664-0.803) in the training set and 0.734 (95% confidence interval 0.608-0.861) in the test set. In the training set, the radiomics signature yielded an AUC of 0.707, 0.737, 0.788, 0.859 and 0.829 for 1-, 2-, 3-, 4- and 5-year PFS, respectively. Similarly, the radiomics signature exhibited an AUC of 0.738, 0.807, 0.761, 0.787 and 0.818 for 1-, 2-, 3-, 4- and 5-year PFS, respectively, in the test set. The calibration curves showed no significant difference between the predicted probability of the radiomics signature and the observed probability for up to 5 years. CONCLUSIONS: Computed tomography radiomics features exhibit a significant correlation with the PFS of neuroblastoma patients, particularly in terms of long-term outcomes.

[Association of lifestyle and cardiometabolic risk factors with epigenetic age acceleration in adults in China].

Objective: To explore the association of lifestyle and cardiometabolic risk factors with five epigenetic age acceleration (AA) indices. Methods: This study included 980 participants of China Kadoorie Biobank, for whom genome-wide DNA methylation of peripheral blood cells had been detected in baseline survey. Five indices of DNA methylation age (DNAm age) were calculated, i.e. Horvath clock, Hannum clock, DNAm PhenoAge, GrimAge and Li clock. Epigenetic AA was defined as the residual of regressing DNAm age on chronological age. Lifestyle factors studied included smoking status, alcohol consumption, eating habits, physical activity level and body shape defined by a combination of BMI and waist circumference. Cardiometabolic risk factors included blood pressure, blood glucose level and total cholesterol level. Linear regression model was used to analyze the association of lifestyle and cardiometabolic risk factors with AA (ß). Results: GrimAge_AA was associated with smoking status, alcohol consumption, physical activity level and BMI. Compared with non-smokers, non-drinkers, or participants with BMI of 18.5- 23.9 kg/m2, the smokers who smoked 1-14 cigarettes/day (ß=0.71, 95%CI: 0.57-0.86), 15-24 cigarettes/day (ß=0.88, 95%CI: 0.73-1.03), and ≥25 cigarettes/day (ß=0.99, 95%CI: 0.81-1.18), respectively, heavy drinkers with daily pure alcohol consumption ≥60 g (ß=0.33, 95%CI: 0.11-0.55) and participants with BMI<18.5 kg/m2 (ß=0.23, 95%CI: 0.03-0.43) showed accelerated aging. Compared with those in the lowest quintile of physical activity level, participants in the top three quintile of physical activity level showed decelerated aging (ß=-0.13, 95%CI: -0.26-0.01, ß=-0.12, 95%CI: -0.26-0.02, and ß=-0.14, 95%CI: -0.27- -0.00, respectively). GrimAge_AA decreased with the increase of the number of healthy lifestyle factors (P<0.001). Compared with the participants with 0 to 1 healthy lifestyle factor, the ß of those with 2, 3, or 4 to 5 healthy lifestyle factors were -0.30 (95%CI: -0.47- -0.12), -0.47 (95%CI: -0.65- -0.30) and -0.72 (95%CI: -0.90- -0.53), respectively. The other four indices were not statistically significantly associated with most lifestyle factors. None of the five indices of AA was associated with blood pressure, blood glucose level or total cholesterol level. Conclusion: People with unhealthy lifestyle showed accelerated epigenetic aging, that is, the predicted DNAm age is older than their own chronological age.

[Factors associated with the outcome of maxillary distraction osteogenesis in cleft lip and palate patients].

Since distraction osteogenesis had been used in maxillofacial surgery in the 1990s,it has become an important option for treating maxillary hypoplasia in cleft lip and palate (CLP) patients. Numerous domestic and international studies reported that the prognosis of distraction osteogenesis might be affected by various factors, including the design of osteotomy line, type of distractor, advancement of maxillary bone and the age of patients, etc. Therefore, the present article reviewed possible impact factors that affect the effectiveness and outcomes of maxillary distraction osteogenesis of CLP from three aspects: treatment methods, type of patients and adjuvant therapy, which hopefully might benefit the formulation of treatment strategies in clinical work and improve the prognosis of distraction osteogenesis.