RNF113A as a poor prognostic factor promotes metastasis and invasion of cervical cancer through miR197/PRP19/P38MAPK signaling pathway.

It has been discovered that aberrant expression of RNF113A plays a significant role in various diseases, including esophageal cancer, hepatocellular carcinoma, and X-linked trichothiodystrophy syndrome. Nevertheless, its functional implications in cervical cancer (CC) remain unclear. The objective of this study was to investigate the role of RNF113A in both the development and prognosis of CC. To achieve this objective, a total of sixty cases were included in the follow-up investigation. The findings revealed a significant up-regulation of RNF113A protein in CC tissues compared to paired paracancerous tissues, and a high expression level of RNF113A was strongly associated with malignant phenotypes such as lymph node metastasis, differentiation degree, depth of invasion, and FIGO stage. Meanwhile, RNF113A was found to be an independent prognostic risk factor, with its high expression significantly correlating with a reduced overall survival period in patients. To elucidate the underlying cause and mechanism of the unfavorable prognosis associated with RNF113A, comprehensive functional investigations were conducted both in vitro and in vivo.Interestingly, it was revealed that RNF113A promoted migration and invasion while inhibiting apoptosis of CC cells, thereby contributing to a poor prognosis. Mechanistically, RNF113A regulated the progression and prognosis of CC through the miR197/Prp19/p38Mark signaling pathway. Overall, our findings underscore the potential clinical significance of RNF113A as an unfavorable prognostic factor in CC.

RNF113A targeted by miR-197 promotes proliferation and inhibits autophagy via CXCR4/CXCL12/AKT/ERK/Beclin1 axis in cervical cancer.

Ring Finger Protein 113 (RNF113A), an ubiquitin E3 ligase, is genetically associated with many biological processes, including proliferation, differentiation, cell death, and neurogenesis. Recently, RNF113A has been found to be an abnormal expression in many diseases, such as X-linked trichothiodystrophy syndrome and esophageal cancer. Here, we explore the potential mechanism of RNF113A in the progression of cervical cancer (CC). In this study, we evaluated the expression level and biological function of RNF113A in CC both in vitro and in vivo by bioinformatic prediction, DIA proteomic analysis, compensation experiment, Co-IP, dual-luciferase reporter assay and nude mouse xenograft to identify the RNF113A-associated autophagy pathways involved with tumorigenesis. Consistent with the prediction from biological information analysis, we found that RNF113A was highly expressed in human CC tissues and cells. In addition, this study illustrated that the high expression of RNF113A dramatically promoted proliferation and suppressed autophagy both in vitro and in vivo. In contrast, low expression of RNF113A enhanced autophagy activities and inhibited tumor growth in CC. We also found that miRNA-197, the level of which (negative correlation with RNF113A) declined in human CC, directly restrained the expression of RNF113A. Mechanistically, proteomic and mechanistic assays uncovered that RNF113A confirmed as the direct downstream target of miR-197, promoted proliferation and restrained autophagy in CC not through direct ubiquitination degradation of autophagy marker Beclin1 but via CXCR4/CXCL12/AKT/ERK/Beclin1 signal transduction axis. In summary, we found a new miR-197/RNF113 A/CXCR4/CXCL12/AKT/ERK/Beclin1 regulation pathway that plays an important part in the survival and progression of CC.

Establishing a differential diagnosis model between primary membranous nephropathy and non-primary membranous nephropathy by machine learning algorithms.

CONTEXT: Four algorithms with relatively balanced complexity and accuracy in deep learning classification algorithm were selected for differential diagnosis of primary membranous nephropathy (PMN). OBJECTIVE: This study explored the most suitable classification algorithm for PMN identification, and to provide data reference for PMN diagnosis research. METHODS: A total of 500 patients were referred to Luo-he Central Hospital from 2019 to 2021. All patients were diagnosed with primary glomerular disease confirmed by renal biopsy, contained 322 cases of PMN, the 178 cases of non-PMN. Using the decision tree, random forest, support vector machine, and extreme gradient boosting (Xgboost) to establish a differential diagnosis model for PMN and non-PMN. Based on the true positive rate, true negative rate, false-positive rate, false-negative rate, accuracy, feature work area under the curve (AUC) of subjects, the best performance of the model was chosen. RESULTS: The efficiency of the Xgboost model based on the above evaluation indicators was the highest, which the diagnosis of PMN of the sensitivity and specificity, respectively 92% and 96%. CONCLUSIONS: The differential diagnosis model for PMN was established successfully and the efficiency performance of the Xgboost model was the best. It could be used for the clinical diagnosis of PMN.

Membranous nephropathy (MN) without obvious causes is called primary MN (PMN), This study utilized deep learning classification algorithms for differential diagnosis of PMN and explored the most suitable classification algorithm for PMN recognition, provided data reference for PMN diagnosis research.

Comparison of heterologous prime-boost immunization strategies with DNA and recombinant vaccinia virus co-expressing GP3 and GP5 of European type porcine reproductive and respiratory syndrome virus in pigs.

Vaccination is principally used to control and treat porcine reproductive and respiratory syndrome virus (PRRSV) infection. This study investigated immunogenicity and protective efficacy of heterologous prime-boost regimens in pigs, including recombinant DNA and vaccinia virus vectors coexpressing PRRSV European genotype (EU) isolate GP3 and GP5: group A, pVAX1-EU-GP3-GP5 prime and rddVTT-EU-GP3-GP5 boost; group B, rddVTT-EU-GP3-GP5 prime and pVAX1-EU-GP3-GP5 boost; group C, empty vector pVAX1; group D, E3L gene-deleted vaccinia virus E3L- VTT. Vaccine efficacy was tested in an EU-type PRRSV (Lelystad virus strain) challenge pig model based on evaluating PRRSV-specific antibody responses, neutralizing antibodies, cytokines, T lymphocyte proliferation, CD4+ and CD8+ T lymphocytes, clinical symptoms, viremia and tissue virus loads. Plasmid DNA was delivered as chitosan-DNA nanoparticles, and Quil A (Quillaja) was used to increase vaccine efficiency. All piglets were boosted 21 days post the initial inoculation (dpi) and then challenged 14 days later. At 14, 21, 28 and 35 dpi, groups A and B developed significantly higher PRRSV-specific antibody responses compared with control groups C and D. Two weeks after the boost, significant differences in neutralizing antibody and IFN-γ levels were observed between groups A, C, D and B. At 49 dpi, groups A and B had markedly increased peripheral blood CD3+CD4+ T cell levels. Following virus challenge, group A showed viremia, but organ virus loads were lower than those in other groups. Thus, a heterologous prime-boost vaccine regimen (rddVTT-EU-GP3-GP5 prime, pVAX1-EU-GP3-GP5 boost) can improve humoral- and cell-mediated immune responses to provide resistance to EU-type PRRSV infection in vivo.

The anti-inflammatory activity of resveratrol in acute kidney injury: a systematic review and meta-analysis of animal studies.

CONTEXT: Accumulated experimental evidence suggests that resveratrol (RSV) may have an effect on acute kidney injury (AKI) by inhibiting inflammation. However, the credibility of the evidence for this practice is unclear. OBJECTIVE: This study investigated the effect of RSV on AKI and the underlying mechanism. METHODS: We searched PubMed, EMBASE, and Web of Science from 2005 to April 2022 for controlled animal trials assessing the effect of conventional resveratrol versus placebo on renal function outcome after AKI. This study was registered within the International Prospective Register of Systematic Reviews (PROSPERO) as number CRD42022329596. RESULTS: We retrieved 455 studies, 25 studies comprising data of 436 animals that met the inclusion criteria. Our meta-analysis suggested that RSV treatment was significantly associated with lower levels of serum creatinine (Scr) and blood urea nitrogen (BUN). The greatest effects were recorded in low-dose (<20 mg/kg/day) groups rather than in high-dose (> 20 mg/kg/day) groups. For time-response effects, subgroup analysis indicated that intervention duration of RSV can influence the treatment effect, and more beneficial effects were observed when studies had a drug administration time of <2 weeks. DISCUSSION AND CONCLUSIONS: This systematic review of animal AKI studies showed a consistently favourable effect of RSV as compared to placebo on renal function outcomes that increased with lower TNF-α, IL-6, and IL-1ß. RSV has a more beneficial effect on SA-AKI animal models than the others. When the RSV intervention dose was low (< 20 mg/kg/day) and the intervention time was <2 weeks, more benefits could be observed.

Epidemiological Investigation and Risk Factors for Cervical Lesions: Cervical Cancer Screening Among Women in Rural Areas of Henan Province China.

BACKGROUND This cross-sectional study aimed to investigate the prevalence of cervical lesions and evaluate risk factors for cervical intraepithelial neoplasia (CIN) among women taking part in cervical cancer screening in rural areas of Henan province, China. MATERIAL AND METHODS Cervical cancer screening using the ThinPrep cytologic test (TCT) and gynecologic exam was conducted on 1315 women age 20-68 years in rural areas of Henan province, China. Colposcopy and biopsies were carried out for histopathologic diagnosis when indicated. Univariate and multivariate logistic regressions were performed to evaluate risk factors associated with cervical lesions. RESULTS Among 1315 women screened, CIN prevalence detected by histopathology was 1.22% (0.38% of CIN 1, 0.76% of CIN 2, and 0.08% of CIN 3). Cervical cancer prevalence was 2.66%. Multivariate analysis confirmed risk factors for cervical lesions included older age (the 21-40 age group vs. the 41-66 age group, OR=0.13, 95% CI: 0.03~0.57), postmenopause (OR=0.11, 95% CI: 0.03~0.45), cervical inflammation (OR=0.06, 95% CI: 0.01~0.31), and smoking (OR=6.78, 95% CI: 1.20~38.23). CONCLUSIONS Older age (41-66 years), presence of HPV infection, postmenopause, cervical inflammation, and smoking are strong risk factors for cervical lesions among women in rural areas of Henan province, China. Particular efforts should be made to provide cervical cancer screening for these women.

[Therapeutic effect of hysteroscopic surgery in treatment of small mass of caesarean scar pregnancy].

OBJECTIVE: Study the effect of hysteroscopic surgery in treatment of small mass of caesarean scar pregnancy (CSP). METHODS: From January 2000 to January 2013, 61 cases lesions ≤ 2.5 cm diameter of endogenous CSP undergoing treatment in Luohe Central Hospital were studied retrospectively. According to different treatment, they were divided into 21 cases in hysteroscopic surgery group, 23 cases in methotrexate+operation group and 17 cases in bilateral uterine artery embolism+curettage group.Intraoperative blood loss, operative time, hospitalization time after treatment and ß-hCG to normal time were compared among those three groups. RESULTS: The surgical blood loss, hospitalization time after treatment, ß-hCG to normal time were (49.8 ± 6.2) ml, (3.5 ± 0.8) days, (21.2 ± 2.4) days in hysteroscopic group, (87.0 ± 30.5) ml, (12.5 ± 1.0) days, (29.6 ± 2.2) days in methotrexate+operation group, the difference was statistically significant (P < 0.05). The operation time were (33 ± 4) minutes in hysteroscopic surgery group and (35 ± 6) minutes in methotrexate+operation group, which did not reached significant difference (P > 0.05). Length of hospital stay after treatment of hysteroscopic surgery group is less than the bilateral uterine artery embolism+curettage group significantly (P < 0.05).Operation time, surgical bleeding and ß-hCG to normal time had no obvious difference between hysteroscopic surgery and in bilateral uterine artery embolism+curettage group (P > 0.05). CONCLUSION: Hysteroscopy surgery in treatment of small mass endogenous CSP is effective, shorter hospitalization time, quick recovery.

Plate centrifugation enhances the efficiency of polyethylenimine-based transfection and lentiviral infection.

PURPOSE: To propose an efficient, reproducible, and consistent transgenic technology based on plate centrifugation, which is particularly useful for polyethylenimine (PEI) transfection and lentiviral infection. METHODS: We optimized multiple factors that could contribute to transfection efficiency, such as the dosage of the PEI or DNA, the working solution buffer used for diluting the PEI or DNA, the incubation time for the PEI/DNA complexes, and the transfection time. RESULTS: Plate centrifugation led to a 5.46-fold increase in the transfection efficiency of PEI-based transfection while maintaining the cell survival rate. Moreover, the average copy number of viral genes in each genome increased 4.96-fold with plate centrifugation. Plate centrifugation alters the spatial arrangement of the PEI/DNA complexes or lentiviruses, increasing the chances of these complexes or viruses coming into contact with target cells, ultimately resulting in improved transfection or infection efficiency. CONCLUSIONS: We present a protocol based on plate centrifugation for transfection or lentiviral infection that is suitable for genetic modification of primary cells or stem cells.

Related Factors with Vascular Dementia: A Two-Sample Mendelian Randomization Study.

Pathogenesis of vascular dementia (VD) is still unclear, there are currently no effective prevention and treatment methods. We applied Mendelian randomization (MR) using summary statistics from large-scale GWAS of metabolites and VD to reveal the causal effect of metabolites on the VD. One set of genetics instrument was used for analysis, derived from publicly available genetic summary data. Which was 32 single-nucleotide polymorphisms robustly associated with metabolites. Inverse-variance weighted, weighted median method, MR-Egger regression, and MR Pleiotropy RESidual Sum and Outlier test were used for MR analyses. Strong evidence for a positive effect of metabolites, which means N6-threonylcarbamoyladenosine (t6A) on VD was found in inverse-variance weighted (odds ratios [OR]: 0.667, 95% confidence interval [CI]: 0.548-0.812, p < 0.001), MR-Egger (OR: 0.647, 95% CI: 0.458-0.913, p = 0.019), and weighted median (OR: 0.650, 95% CI: 0.466-0.908, p = 0.012). The MR analysis indicated that metabolites (t6A) may be causally associated with a positive effect on VD.

Current research status and clinical applications of noninvasive preimplantation genetic testing: A review.

Noninvasive preimplantation genetic testing (ni-PGT) is conducted by obtaining genetic information from embryos through the analysis of free DNA released by embryos in spent embryo culture medium or blastocoel fluid. Compared to conventional preimplantation genetic testing relying on trophectoderm biopsy, ni-PGT is characterized by its noninvasiveness. It has demonstrated early advancements in the detection of embryonic chromosomal aneuploidies and the diagnosis of monogenic diseases, showcasing considerable potential for clinical application. However, there are substantial controversies in the literature concerning the reliability of ni-PGT, the source of cell-free DNA, and maternal contamination. This paper elaborates on the principles, research advancements, effectiveness, and limitations of ni-PGT to provide a basis for clinical applications.

Analysis of the current state of cervical cancer prevention awareness and its influencing factors among rural women in Luohe City.

The purpose of this cross-sectional study was to identify the current awareness about cervical cancer prevention among rural women in Luohe City as well as its potential influencing factors. Meanwhile, these data were expected to provide a theoretical basis for Luohe future cervical cancer prevention and therapy. Based on geographical distribution, 40 villages in Luohe City were randomly selected, and questionnaires were given to women in each village. In this study, a total of 4665 questionnaires were distributed, and 4561 valid questionnaires were returned, with a recovery rate of 97.98%. The average score was 4.06 ±â€…2.46 out of 10. It was found that women had a high awareness rate of cervical cancer screening (55.25%) but a low awareness rate of human papillomavirus (HPV) and HPV vaccine (10.17%). Moreover, univariate and multivariable analyses showed that age > 45 years, low household income, low education level, being a farmer, spouse unemployment, no pregnancy or birth delivery history, no family or personal history of cervical disease, and no previous complimentary 2-cancer screening (i.e., breast cancer and cervical cancer) were all factors influencing the cognitive level of rural women in Luohe City (P < .05). However, ethnicity, marital status, and spouse education level were not correlated with cognitive level (P > .05). In conclusion, low awareness of cervical cancer prevention among rural women in Luohe was correlated with individual, family, and social factors. So it was recommended to cultivate the rural population knowledge, optimize screening strategies, and conduct targeted cervical cancer prevention and treatment in rural regions.

Short-term effects of exposure to ambient PM2.5 and its components on hospital admissions for threatened and spontaneous abortions: A multicity case-crossover study in China.

BACKGROUND: The adverse effects of short-term exposure to PM2.5 and its components on hospital admissions for threatened and spontaneous abortions (TSAB) are still controversial. METHODS: Data on daily hospitalizations for TSAB and PM2.5 and its components, including sulfate (SO42-), nitrate (NO3-), ammonium salt (NH4+), organic matter (OM), and black carbon (BC), were collected from January 2015 to December 2021 (total 2,557 days) in five cities in China. Case-crossover analyses were conducted to investigate the short-term associations between PM2.5 and its components and TSAB. Additionally, the modification effects by age (<35 and ≥35 years), season (cold and warm seasons), and the "Three-Year Action Plan to Win the Blue Sky Defense War" (before and after implementation) on the above associations were further conducted. RESULTS: For each 10 µg/m3 (1 µg/m3 for BC) increase, the strongest relative risks (95% confidence intervals) of hospitalization for TSAB were 1.011 (1.001-1.021) for PM2.5 in lag02, 1.060 (1.003-1.120) for SO42- in lag02, 1.035 (1.000-1.070) for NO3- in lag02, 1.065 (1.009-1.124) for NH4+ in lag02, 1.047 (1.008-1.088) for OM in lag01 and 1.029 (1.005-1.054) for BC in lag02 (all P <0.05). Furthermore, significant modifying effects of age and the Action Plan were found. The effects of NO3- (lag2), NH4+ (lag2), and BC (lag2) were more pronounced in mothers aged ≥35 years and the effects of PM2.5 (lag4), NO3- (lag4), NH4+ (lag4), OM (lag4), and BC (lag4) was more pronounced in the period before the Action Plan was implemented (all P modification <0.05). CONCLUSION: Short-term exposure to ambient PM2.5 and its components (SO42-, NO3-, NH4+, OM, and BC) was related to increased risks of hospitalization for TSAB. The effects were more pronounced in mothers aged ≥35 years and the period before the Action Plan.

Association between pelvic floor muscle strength and sexual function based on PISQ-12-an analysis of data from a multicenter cross-sectional study on 735 nulliparae during pregnancy.

Background: Pelvic floor muscle strength is well-known to be associated with female sexual function. However, there were a few studies that reported on the relationship between pelvic floor muscle strength and female sexual function in pregnant women, and the presented results were inconsistent. Nulliparae represent a specific cohort with simplicity to exclude confounding factors that are caused by parity. The present study aimed to explore the association of pelvic floor muscle strength and sexual function based on the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12) of nulliparae during pregnancy. Methods: This is the second analysis of the baseline data from a randomized controlled trial (RCT), which aimed to study the protective efficacy of pelvic floor muscle training on stress urinary incontinence at 6th week postpartum (registration number: ChiCTR2000029618). Nulliparae aged 20-40 years with singleton pregnancy before 16 weeks of gestation were enrolled in this study, and data, including participants' demographic information, the Modified Oxford Scale (MOS), and PISQ-12, were collected. Eligible nulliparae were divided into two groups: Group MOS > 3 and Group MOS ≤ 3. Demographic information of the two groups was compared. Sexual function based on the PISQ-12 scores of the two groups was compared. A comparison of the PISQ-12 scores between the two groups was calculated by the Mann-Whitney U-test using SPSS version 23.0. Results: A total of 735 eligible nulliparae were enrolled in this study. Along with MOS grading up, PISQ-12 scores tended to get lower. Of the 735 nulliparae, there were 378 and 357 participants included in Group MOS > 3 and Group MOS ≤ 3, respectively. The PISQ-12 scores of Group MOS > 3 were significantly lower than those of Group MOS ≤ 3 (11 vs. 12, p < 0.001). The scores of the frequency of feeling sexual desire, orgasm achievement, sexual excitement, sexual activity satisfaction, sexual intercourse pain, fear of urinary incontinence, and negative emotion reactions with the sexual intercourse of Group MOS > 3 were lower than those of Group MOS ≤ 3 (p < 0.05). Conclusion: Pelvic floor muscle strength was positively associated with sexual function based on the questionnaire of young nulliparae during their first trimester. Up to half of the nulliparae during the first trimester were suffering from weak pelvic floor muscle strength and nearly a quarter of the nulliparae were facing this weakness combined with sexual dysfunction. Trial registration: This study has been registered at http://www.chictr.org.cn (registration number: ChiCTR2000029618).

Pelvic Floor Muscle Strength in the First Trimester of Primipara: A Cross-Sectional Study.

Background: Pelvic floor muscle (PFM) weakness is associated with stress urinary incontinence. Pregnancy is an important risk factor for PFM weakness. Studies evaluating PFM strength in the first trimester of pregnancy are still lacking. Our study aimed to describe pelvic floor function of the primipara in the first trimester of gestation and investigate the risk factors for PFM weakness. Methods: Primiparas aged 20~40 years with a singleton pregnancy less than 14 weeks of gestation were recruited, and data were collected via questionnaires on items that were suggested as associated with PFM weakness, followed by Modified Oxford Scale (MOS) on genital hiatus and perineal body and pelvic floor ultrasound evaluation for the thickness of the left and right levator ani muscles (LAM), right−left diameter of the levator hiatus (LH), and LH area. Participants were divided into three groups by MOS >3, =3, and <3 for data analysis. Results: A total of 380 participants completed the questionnaires and examinational analysis, of whom, 228, 98, and 54 were divided into Group 1, Group 2, and Group 3, respectively. The three groups were significantly different in the number of gestations and abortions, toilet types, and the right−left diameter of the LH (p < 0.05). Logistic regressive analysis showed that squatting toilet dominant (OR = 3.025; 95% CI: 1.623~5.638; p < 0.001) and a larger right−left diameter of the LH (OR = 1.065; 95% CI: 1.026~1.105; p = 0.001) were significantly associated with PFM weakness. Conclusions: Squatting toilet dominancy and longer right−left diameter of the LH are significantly associated with PFM weakness in primiparas in the first trimester. Sitting toilets should be recommended to women, especially pregnant women. Trial registration: The trial has been registered at Chinese Clinical Trial Registry (registration number: ChiCTR2000029618).

Multiple regression analysis of perinatal conditions, physical development, and complications in assisted reproduction singletons.

BACKGROUND: There is no clear clinical conclusion on whether assisted reproductive technology (ART) increases maternal and child risk and affects infant birth and development. This study aimed to perform a multiple regression analysis of the perinatal maternal and infant conditions in an assisted reproductive singleton pregnancy and the physical development and complications of such infants at 6 months old. METHODS: This study enrolled 145 singleton pregnant women who were admitted to Luohe Central Hospital between December 2017 and December 2019 to undergo in vitro fertilization and embryo transfer as a research group, and 160 singleton pregnant women who were naturally conceived at the same time and delivered at our hospital were selected as the control group. The relevant data of the patients were collected, and the perinatal conditions, neonatal complications, physical development and NBN score of infants aged 6 months were compared between the two groups. Multivariate logistic regression was used to analyze risk factors for perinatal complications in assisted reproduction singleton pregnancy. RESULTS: The incidence of complications in the perinatal period in the study group was significantly higher than that in the control group (P<0.05). However, there was no significant difference in the incidence of complications such as very low birth weight, respiratory distress, neonatal hemolysis, hyperbilirubinemia, hypoglycemia, infectious pneumonia and neonatal asphyxia between the two groups (P>0.05). Likewise, at 6 months of age, the infants in the research group showed no significant difference to the control group in physical development indicators, including weight, head circumference, body mass index, height and NBN score (P>0.05). The analysis of the unconditional multivariate logistic regression model revealed hypertension during pregnancy, placental previa, premature membrane rupture, gestational age <32 weeks, and very low birth weight as risk factors that affect perinatal complications in patients with assisted reproductive singleton pregnancy (P<0.05). CONCLUSIONS: The incidence of complications in perinatal patients with assisted reproductive singleton pregnancy is higher than that of natural singleton pregnancy, but there is no significant difference in physical development, NBN score and complications of 6 months old infants.

[Incidence and risk factors of stroke in patients with nonvalvular atrial fibrillation in Haikou].

OBJECTIVE: To investigate the incidence of ischemic stroke and associated risk factors in patients with nonvalvular atrial fibrillation (NVAF) in Haikou. METHODS: Area through follow-up approach and evaluate the effect of antithrombotic therapy as well as the current status of treatment in patients with NVAF. Clinical data of 329 NVAF patients admitted to Hainan hospital from January 1999 to December 2001 were collected. The patients were followed up to identify the incidence of ischemic stroke and associated risk factors. RESULTS: The main cause of NVAF was coronary heart diseases, which happened in 207 cases (62.9%). The logistic regression analysis revealed that age, type of atrial fibrillation, and hypertension were significantly associated with the occurrence of ischemic stroke in patients with NVAF (P=0.0001, 0.006, and 0.04 respectively). Antithrombotic treatments significantly decreased the incidence of stroke (P<0.05). But Warfarin was not applied consistently in Haikou. CONCLUSIONS: Age, type of atrial fibrillation and hypertension are independent risk factors for ischemic stroke in patients with NVAF. Uninterrupted use of aspirin can be beneficial to patients with NVAF.

Association of clinical outcomes in metastatic breast cancer patients with circulating tumour cell and circulating cell-free DNA.

BACKGROUND: Both circulating tumour cell (CTC) and total circulating cell-free DNA (ccfDNA) predict cancer patient prognosis. However, no study has explored the prognostic value of the combined use of CTC and ccfDNA. We aimed to investigate individual and joint effects of CTC and ccfDNA on clinical outcomes of metastatic breast cancer (MBC) patients. METHODS: We collected 227 blood samples from 117 MBC patients. CTCs were enumerated using the CellSearch System. ccfDNAs were quantified by quantitative real-time polymerase chain reaction and Qubit fluorometer. The individual and joint effects of CTC and ccfDNA levels on patient progression-free survival (PFS) and overall survival (OS) were analysed using Cox proportional hazards models. RESULTS: Compared to patients with <5 CTCs, patients with ≥5 CTCs had a 2.58-fold increased risk of progression and 3.63-fold increased risk of death. High level of ccfDNA was associated with a 2.05-fold increased risk of progression and 3.56-fold increased risk of death. These associations remained significant after adjusting for other important clinical covariates and CTC/ccfDNA levels. CTC and ccfDNA levels had a joint effect on patient outcomes. Compared to patients with low levels of both CTC and ccfDNA, those with high levels of both markers exhibited a >17-fold increased death risk (P < 0.001). Moreover, longitudinal analysis of 132 samples from 22 patients suggested that the inconsistency between CTC level and outcome in some patients could possibly be explained by ccfDNA level. CONCLUSIONS: CTC and total ccfDNA levels were individually and jointly associated with PFS and OS in MBC patients.

Polymorphism of IFN-γ +874T/A associated with production of IFN-γ affects human papillomavirus susceptibility in rural women from Luohe, Henan, China.

PURPOSE: In this paper, the association between polymorphisms of IFN-γ +874T/A (rs2430561), IFN-γR1 -56 T/C (rs2234711), IFN-γR1 +95 C/T (rs7749390), and IFN-γR1 -611A/G (rs 1327474) and human papillomavirus (HPV) susceptibility was investigated in rural women from Luohe, Henan, China. PATIENTS AND METHODS: A total of 520 rural women were enrolled from Luohe, including 260 with HPV infection and mild dysplasia or less and 260 without HPV infection. Single-nucleotide polymorphisms (SNPs) of IFN-γ +874T/A, IFN-γR1 -56 T/C, IFN-γR1 +95 C/T and IFN-γR1 -611A/G were genotyped using TaqMan Pre-Designed SNP Genotyping Assays. Serum IFN-γ levels were measured using Human IFN-γ Quantikine ELISA Kit. Multivariate logistic regression analysis was performed to identify the SNPs associated with HPV susceptibility. Serum IFN-γ levels were compared between different genotypes. RESULTS: The polymorphism of IFN-γ +874T/A was associated with HPV susceptibility and +874A carriers had an increased risk. Moreover, the odds ratio was higher in +874 AA carriers than in +874 AT carriers (1.672 vs 2.874). Serum IFN-γ levels were highest in IFN-γ +874 TT carriers, intermediate in AT carriers, and lowest in AA carriers (2.86±1.14 vs 1.57±0.79 vs 0.41±0.22 pg/mL, all P<0.05). CONCLUSION: The polymorphism of IFN-γ +874T/A was associated with HPV susceptibility in rural women from Luohe, Henan, China, and +874A carriers had an increased risk. The possible mechanism was that +874A carriers had a low production of IFN-γ.

Optimized PEI-based Transfection Method for Transient Transfection and Lentiviral Production.

Polyethyleneimine (PEI), a cationic polymer vehicle, forms a complex with DNA which then can carry anionic nucleic acids into eukaryotic cells. PEI-based transfection is widely used for transient transfection of plasmid DNA. The efficiency of PEI-based transfection is affected by numerous factors, including the way the PEI/DNA complex is prepared, the ratio of PEI to DNA, the concentration of DNA, the storage conditions of PEI solutions, and more. Considering the major influencing factors, PEI-based transfection has been optimized to improve its efficiency, reproducibility, and consistency. This protocol outlines the steps for ordinary transient transfection and lentiviral production using PEI. © 2017 by John Wiley & Sons, Inc.

Simulated ischemia/reperfusion-induced p65-Beclin 1-dependent autophagic cell death in human umbilical vein endothelial cells.

Myocardial ischemia/reperfusion (I/R) injury detrimentally alters the prognosis of patients undergoing revascularization after acute myocardial infarction. Our previous study demonstrated that NF-κB-induced autophagy plays a detrimental role in cardiac I/R injury using a rabbit myocardial I/R model. In this study, we sought to explore the specific mechanism of this autophagy-mediated cell damage in an in vitro simulated ischemia/reperfusion (sI/R) model using human umbilical vein endothelial cells. Our current study demonstrates that simulated I/R induces autophagy in a p65-Beclin 1-dependent manner, which can be suppressed with the inhibition of NF-κB. Furthermore, rapamycin which promotes autophagy, exacerbates sI/R-induced cell death. While 3-methyladenine rescues cell damage. Our data thus suggest that I/R promotes NF-κB p65 activity mediated Beclin 1-mediated autophagic flux, thereby exacerbating myocardial injury.