PGAM5 degrades PDCoV N protein and activates type I interferon to antagonize viral replication.

IMPORTANCE: As a member of the δ-coronavirus family, porcine deltacoronavirus (PDCoV) is a vital reason for diarrhea in piglets, which can contribute to high morbidity and mortality rates. Initially identified in Hong Kong in 2012, the virus has rapidly spread worldwide. During PDCoV infection, the virus employs evasion mechanisms to evade host surveillance, while the host mounts corresponding responses to impede viral replication. Our research has revealed that PDCoV infection down-regulates the expression of PGAM5 to promote virus replication. In contrast, PGAM5 degrades PDCoV N through autophagy by interacting with the cargo receptor P62 and the E3 ubiquitination ligase STUB1. Additionally, PGAM5 interacts with MyD88 and TRAF3 to activate the IFN signal pathway, resulting in the inhibition of viral replication.

Mitochondrial KATP channel-mediated autophagy contributes to angiotensin II-induced vascular dysfunction in mice.

BACKGROUND AND AIM: The present study aimed to investigate whether the mitochondrial KATP channel contributes to angiotensin II (Ang II)-induced vascular dysfunction, the development of hypertension, and atherosclerosis. METHODS AND RESULTS: ApoE (-/-) mice fed a high-fat diet were chronically infused with Ang II for eight weeks and concomitantly treated with losartan (ARB), apocynin, or 5-hydroxy decanoate (5-HD), or 3-methyladenine (3-MA). Systolic blood pressure was measured, and pathological changes of aortic or liver tissue were observed. Nitric oxide (NO), superoxide dismutase 2 (SOD2) levels and vasorelaxation rate were measured, and protein and mRNA expressions were examined by western blot and RT-PCR. Ang II-induced development of hypertension was suppressed not only by ARB, and apocynin but also by 5-HD or 3-MA. Ang II infusion decreased aortic NO production and relaxation, as well as SOD2 activity in liver, which were improved by all treatments. In addition, Ang II-induced activation of autophagy was suppressed by 5-HD in aortic tissue, furthermore, Ang II increases the atherosclerotic index in plasma and exacerbates the development of atherosclerosis by increases of fat deposition in the aorta and liver. Lipid metabolism-related mRNA expressions (LXR-α, LDLR, SRBI, Acca, and FASN) were changed by Ang II. Similarly, not only ARB, and apocynin, but also 5-HD and 3-MA suppressed Ang II-induced these changes. CONCLUSIONS: Our present findings evidence that mitochondrial KATP channel-mediated autophagy contributes to Ang II-induced vascular dysfunction, development of hypertension, and atherosclerosis.

Efficacy and safety of 755-nm picosecond alexandrite laser with topical tranexamic acid versus laser monotherapy for melasma and facial rejuvenation: a multicenter, randomized, double-blinded, split-face study in Chinese patients.

To compare the efficacy and safety of 755-nm picosecond alexandrite laser and topical tranexamic acid (TTA) combination therapy with laser monotherapy, for the treatment of melasma and facial rejuvenation. This multicenter, randomized, double-blinded, split-face study enrolled 37 patients who presented with melasma and photoaging. Facial halves were randomized to receive either laser and TTA combination therapy or laser monotherapy. Three treatments were delivered at 4-5 weeks intervals. Patients were followed up for 1, 3, and 6 months post-final treatment and evaluated by blinded investigators for hemi-Melasma Area and Severity Index (hemi-MASI), facial dyschromia, skin texture, laxity, and rhytids. Daily diaries rating healing progress for 7 days posttreatment and satisfaction grading were performed by all patients. Adverse events were recorded. Thirty-six patients completed the follow-up. Compared with the baseline, hemi-MASI, dyschromia, and skin texture on both halves improved significantly through the follow-up (p = 0.000). A significant difference in hemi-MASI and dyschromia between combination therapy halves and monotherapy halves was noticed at 1- and 3-month follow-ups (p < 0.05). The laser monotherapy halves displayed significantly less redness and sensitivity during the 7-day posttreatment recovery period (p < 0.05). Patients' satisfaction ratings for the combination therapy halves were higher than the monotherapy halves at 1-month follow-up (p < 0.05). No severe adverse events were observed. The picosecond alexandrite laser and TTA combination therapy demonstrated synergistic efficacy for hemi-MASI and dyschromia improvements over laser monotherapy. The optimization of the picosecond laser and TTA combination regimen needs further investigation.

Expression pattern Nanog is associated with the development of gestational trophoblastic neoplasia in Chinese women from Fujian province: a case-control study.

BACKGROUND: Gestational trophoblastic disease (GTD) is a group of interrelated but distinct diseases and has a serious impact on the reproductive health of women. To analyze the expression of Nanog in GTD and to evaluate its potential to predict the development of gestational trophoblastic neoplasia (GTN). METHODS: The study included 41 normal first-trimester placentas matched by gestational age to 53 regressed-hydatidiform-moles (rHMs), 56 malignant-HMs (mHMs) and 17 choriocarcinomas (CCAs) and evaluated the Nanog expression by immunohistochemistry. The chi-square test, ANOVA, Fisher's exact test and logistic regression were performed to assess the Nanog expression and clinical prognostic factors in GTD. RESULTS: Compared to normal placenta levels, the Nanog expression was increased in GTD samples (p < .05). In HMs, Nanog expression was positively correlated with serum ß-hCG levels,uterine size and theca-lutein cysts (p < .05). Compared with the low-risk metastatic group (Federation of Gynecology and Obstetrics (FIGO) score ≤ 6), the high-risk metastatic group (FIGO score >7) had higher Nanog expression (p = .030). Moreover, logistic regression analysis showed that the positive expression of Nanog had the highest risk of developing into GTN (OR = 4.764, p < .001). CONCLUSIONS: Nanog is an independent predictor of clinical outcomes. It can also be a reliable predictor for GTN development from GTD.

Knowledge and Attitudes Regarding HPV and Vaccination Among Chinese Women Aged 20 to 35 Years in Fujian Province: A Cross-Sectional Study.

The use of the human papillomavirus (HPV) vaccine was recently approved in Mainland China. This study determined the knowledge and attitudes of young women aged 20 to 35 years in Fujian Province, China, with regard to HPV and vaccination and explored the potential factors influencing their attitudes toward HPV vaccination. This was a cross-sectional study that collected data regarding the knowledge on and attitudes toward HPV and vaccination using questionnaires. Furthermore, the prevalence of HPV was determined from the sampled participants. A total of 1001 young women were included in the survey. This study demonstrated that the HPV prevalence rate was 15.7% (157/1001). Among all patients, 44.9% (n = 449) had heard of HPV; however, detailed knowledge about HPV was lacking. The majority (83.7%) expressed a willingness to be vaccinated. Specifically, knowledge of the dangers of HPV infection was significantly associated with the willingness to be vaccinated. In this study, women cited some concerns and expressed high expectations for the HPV vaccine, but the costs of vaccination reduced their willingness to be vaccinated. This study found that most patients did not have a detailed knowledge of HPV. Thus, there is a need for continued HPV promotion and education efforts, especially on the dangers of HPV infection, among young women aged 20 to 35 years in Fujian Province, China. Furthermore, it is important to subsidize the costs of vaccination for promoting vaccination campaigns in China.

Comprehensive view of macrophage autophagy and its application in cardiovascular diseases.

Cardiovascular diseases (CVDs) are the primary drivers of the growing public health epidemic and the leading cause of premature mortality and economic burden worldwide. With decades of research, CVDs have been proven to be associated with the dysregulation of the inflammatory response, with macrophages playing imperative roles in influencing the prognosis of CVDs. Autophagy is a conserved pathway that maintains cellular functions. Emerging evidence has revealed an intrinsic connection between autophagy and macrophage functions. This review focuses on the role and underlying mechanisms of autophagy-mediated regulation of macrophage plasticity in polarization, inflammasome activation, cytokine secretion, metabolism, phagocytosis, and the number of macrophages. In addition, autophagy has been shown to connect macrophages and heart cells. It is attributed to specific substrate degradation or signalling pathway activation by autophagy-related proteins. Referring to the latest reports, applications targeting macrophage autophagy have been discussed in CVDs, such as atherosclerosis, myocardial infarction, heart failure, and myocarditis. This review describes a novel approach for future CVD therapies.

Exosomes based strategies for cardiovascular diseases: Opportunities and challenges.

Exosomes, as nanoscale extracellular vesicles (EVs), are secreted by all types of cells to facilitate intercellular communication in living organisms. After being taken up by neighboring or distant cells, exosomes can alter the expression levels of target genes in recipient cells and thereby affect their pathophysiological outcomes depending on payloads encapsulated therein. The functions and mechanisms of exosomes in cardiovascular diseases have attracted much attention in recent years and are thought to have cardioprotective and regenerative potential. This review summarizes the biogenesis and molecular contents of exosomes and details the roles played by exosomes released from various cells in the progression and recovery of cardiovascular disease. The review also discusses the current status of traditional exosomes in cardiovascular tissue engineering and regenerative medicine, pointing out several limitations in their application. It emphasizes that some of the existing emerging industrial or bioengineering technologies are promising to compensate for these shortcomings, and the combined application of exosomes and biomaterials provides an opportunity for mutual enhancement of their performance. The integration of exosome-based cell-free diagnostic and therapeutic options will contribute to the further development of cardiovascular regenerative medicine.

Intervention with ICOSL Antibodies Alleviates Inflammatory Infiltrations in Mice with Neutrophilic Asthma.

Background: Neutrophilic asthma is characterized by the predominant infiltration of neutrophils in airway inflammation. Objective: To explore the therapeutic potential of an antibody against the inducible T cell co-stimulator ligand (ICOSL) in a mouse model of neutrophilic asthma. Methods: Female BALB/c mice were randomly assigned to different groups. They were then injected with ovalbumin (OVA)/lipopolysaccharides (LPS) to induce neutrophilic asthma. The mice were then treated with either anti-ICOSL (the I group), control IgG (the G group), or no treatment (the N group). Additionally, a control group of mice received vehicle PBS and was labeled as the C group (n=6 per group). One day after the last allergen exposure, cytokine levels were measured in plasma and bronchoalveolar lavage fluid (BALF) using ELISA. After analyzing and categorizing BALF cells, the lung tissues were examined histologically and immunohistochemically. Results: Administering anti-ICOSL resulted in a significant decrease in the total number of inflammatory infiltrates and neutrophils found in BALF. Moreover, it led to a decrease in the levels of interleukin (IL)-6, IL-13, and IL-17 in both BALF and plasma. Additionally, there was an increase in IFN-γ levels in the BALF of asthmatic mice (p<0.05 for all). Treatment with anti-ICOSL also reduced lung interstitial inflammation, mucus secretion, and ICOSL expression in asthmatic mice. Conclusion: The treatment of anti-ICOSL effectively improved lung interstitial inflammation and mucus secretion in mice with neutrophilic asthma by restoring the balance of Th1/Th2/Th17 responses. These findings indicate that blocking the ICOS/ICOSL signaling could be an effective way to manage neutrophilic asthma.

[Expression and significance of matriptase in ovarian cancer cells with diverse metastatic potential].

OBJECTIVE: To study the expression and significance of matriptase in different metastatic potential of human ovarian cancer cells. METHODS: High-metastatic human ovarian cancer cell HO8910PM and ovarian cancer cell HO8910 were collected.The ability of metastatic of the former was stronger than that of the latter. Compared the ability of invasion and migration in HO8910PM and HO8910 by scratch assay and by millicell chamber artificial reconstituted basement membrane invasion assay. Detected the matriptase mRNA and protein expression levels in HO8910PM and HO8910 through reverse transcription(RT)-PCR and immunocytochemistry methods. RESULTS: The 24 hours' migration distance (347 ± 8) µm of HO8910PM cells were significantly higher than that in HO8910 group (154 ± 10) µm (P < 0.01);The number of HO8910PM cells that penetrated the matrigel after 24 hours' incubation were significantly higher than that in HO8910 group (90.7 ± 2.1 vs 63.3 ± 1.5,P < 0.01). The expression of matriptase mRNA in HO8910PM cells was higher than that in HO8910 group (0.72 ± 0.03 vs 0.38 ± 0.04,P < 0.01). The migration was positively correlated with the matriptase mRNA expression levels (r = 0.992, P < 0.01); and the invasiveness was also positively correlated with the matriptase mRNA expression levels (r = 0.973, P < 0.01). As far protein level,the expression of matriptase protein in HO8910PM cells was higher than that in HO8910 group (15.6 ± 0.8 vs 7.6 ± 1.3,P < 0.01). The migration was positively correlated with matriptase protein expression levels (r = 0.971, P < 0.01); And the invasiveness was also positively correlated with the matriptase protein expression levels (r = 0.958, P < 0.01). CONCLUSIONS: The relationship between the expression levels of matriptase and the metastatic of ovarian cancer cells may be correlative. The function of matriptase in ovarian cancer cells metastatic mechanism still need to be confirmed.

N protein of PEDV plays chess game with host proteins by selective autophagy.

Macroautophagy/autophagy is a cellular degradation and recycling process that maintains the homeostasis of organisms. The protein degradation role of autophagy has been widely used to control viral infection at multiple levels. In the ongoing evolutionary arms race, viruses have developed various ways to hijack and subvert autophagy in favor of its replication. It is still unclear exactly how autophagy affects or inhibits viruses. In this study, we have found a novel host restriction factor, HNRNPA1, that could inhibit PEDV replication by degrading viral nucleocapsid (N) protein. The restriction factor activates the HNRNPA1-MARCHF8/MARCH8-CALCOCO2/NDP52-autophagosome pathway with the help of transcription factor EGR1 targeting the HNRNPA1 promoter. HNRNPA1 could also promote the expression of IFN to facilitate the host antiviral defense response for antagonizing PEDV infection through RIGI protein interaction. During viral replication, we found that PEDV can, in contrast, degrade the host antiviral proteins HNRNPA1 and others (FUBP3, HNRNPK, PTBP1, and TARDBP) through its N protein through the autophagy pathway. These results reveal the dual function of selective autophagy in PEDV N and host proteins, which could promote the ubiquitination of viral particles and host antiviral proteins and degradation both of the proteins to regulate the relationship between virus infection and host innate immunity.Abbreviations: 3-MA: 3-methyladenine; ATG: autophagy related; Baf A1: bafilomycin A1; CALCOCO2/NDP52: calcium binding and coiled-coil domain 2; ChIP: chromatin immunoprecipitation; Co-IP: co-immunoprecipitation; CQ: chloroquine; DAPI: 4',6-diamidino-2-phenylindole; GPI: glycosyl-phosphatidylinositol; hpi: hours post infection; MARCHF8/MARCH8: membrane-associated ring-CH-type finger 8; MOI: multiplicity of infection; N protein: nucleocapsid protein; PEDV: porcine epidemic diarrhea virus; siRNA: small interfering RNA; TCID50: 50% tissue culture infectious doses.

[Significance of urodynamic analysis in the patients with recent voiding dysfunction after radical hysterectomy].

OBJECTIVE: To study the urodynamic changes in patients with recent non-infective voiding dysfunction following radical hysterectomy and assess its significance. METHODS: Ninety-six patients with cervical cancer, who were not found any abnormal representation of urodynamics before the operation, were selected into this study group. Eighty-three patients in the study group without urinary infection were detected by urodynamic examination following radical hysterectomy, in order to analyze the urodynamic reasons for the non-infective voiding dysfunction following the surgery. RESULTS: Forty-two patients were found with non-infective voiding dysfunction after the operation. Low compliance bladder, bladder destrusor dysfunction and destrusor overactivity were the three leading types of postoperative bladder dysfunction. Moreover, the incidences of low compliance bladder (50.0% vs. 17.1%), bladder destrusor dysfunction (58.4% vs. 14.6%) and destrusor overactivity (31.0% vs. 4.9%) in the group with voiding dysfunction were significantly higher than the corresponding values in the group without voiding dysfunction (P < 0.01). Secondarily, forty-two patients with recent non-infective voiding dysfunction were divided into simple irritation sign group, simple obstruction sign group and mixed sign group according to their main symptoms. The incidence of bladder destrusor dysfunction in the simple obstruction sign group was significant higher than that in the simple irritation sign group, and the incidence of detrusor overactivity in the simple irritation sign group was significant higher than that in the other two groups (P < 0.05). CONCLUSIONS: There were many different types of urodynamic disorder in the patients with recent non-infective voiding dysfunction after radical hysterectomy. Low compliance bladder, bladder destrusor dysfunction and detrusor overactivity caused by the damage of the pelvic autonomic nerve during the operation may be the main reasons for the recent non-infective voiding dusfunction after radical hysterectomy. Moreover, bladder destrusor dysfunction and detrusor overactivity may be the key points for the symptoms of bladder irritation and bladder obstruction. Urodynamic study is important for the etiology analysis and clinical treatment of recent non-infective voiding dysfunction postoperation.

[Influence of urodynamic factors on urinary retention in patients with cervical carcinoma after radical hysterectomy].

OBJECTIVE: To study the effect of urodynamic factors on the urinary retention of the patients with cervical cancer received radical hysterectomy. METHODS: Seventy-two patients with cervical cancer International Federation of Gynecology and Obstetrics (FIGO) stage Ib1 to IIa hospitalized in Fujian Provincial Maternity and Child Health Hospital between June 2006 and August 2009, who were not found any abnormal representation of urodynamics before the operation, were divided into the group with urinary retention and the group without urinary retention based on whether urinary retention after the operation. All patients were detected by urodynamic examination following radical hysterectomy. Data obtained from urodynamic examination were analysed by logistic regression to evaluate the influence of urodynamic factors on the urinary retention postoperation. RESULTS: Twenty-one patients out of all were found with urinary retention after the operation, the incidence rate of urinary retention was 29%. The first sensation after operation in both groups were increased significantly than those before operation [(171 ± 61) ml vs. (126 ± 28) ml, (134 ± 39) ml vs. (119 ± 17) ml, all P < 0.05], while the maximum volume [(337 ± 66) and (300 ± 66) ml, respectively], the compliance [(31 ± 25) and (29 ± 18) ml/cm H2O (1 cm H2O = 0.098 kPa), respectively], the maximum flow rate [(10 ± 4) and (12 ± 5) ml/s, respectively] and the pressure at the maximum flow rate [(27 ± 9) and (32 ± 8) cm H2O, respectively] were decreased obviously after radical hysterectomy in both the group with urinary retention and the group without urinary retention (all P < 0.05), compared with the corresponding value before the operation. The urodynamic changes in urinary retention group was much more severe than those in group without urinary retention (P < 0.05). The single factor analysis results showed that bladder destusor dysfunction (OR = 8.20, 95%CI: 2.62-25.66, P < 0.01) and lack of sensation (OR = 6.90, 95%CI: 1.95-24.43, P < 0.01) were relevant to the urinary retention post-operation. While there were not relationship was found between low compliance bladder (OR = 1.99, 95%CI: 0.70-5.63, P = 0.195), detrusor overactivity (OR = 2.51, 95%CI: 0.73-8.67, P = 0.144), bladder outlet obstruction (OR = 3.77, 95%CI: 0.76-18.57, P = 0.104) or dyssynergia of urethral external sphincter (OR = 2.67, 95%CI: 0.49-14.45, P = 0.255) and urinary retention following the operation. There were an antagonistic effects (OR = 7.60, 95%CI: 1.43-40.39, P = 0.017) of detrusor overactivity and bladder destrusor dysfunction on urinary retention. The multiple factors analysis results revealed that bladder destusor dysfunction (OR = 7.01, P < 0.01) and lack of sensation (OR = 5.45, P = 0.018)were the independent risk factors influencing on the urinary retention post-operation. CONCLUSIONS: There are obvious urodynamic change in cervical cancer patients following radical hysterectomy. Bladder destrusor dysfunction and lack of sensation are the independent urodynamic risk factors influencing on urinary retention following radical hysterectomy, while detrusor over activity may be a protective effect on bladder destrusor dysfunction post-operation in some degree. Urodynamic test is important for analysis and treatment of urinary retention following radical hysterectomy.

Novel turn-on fluorescence sensor for detection and imaging of endogenous H2S induced by sodium nitroprusside.

Currently, fluorescence analysis method has a good application in the detection and imaging of biomarkers and has become an important analytical method. Although there are many fluorescent probes for detecting hydrogen sulfide(H2S), they are mostly based on fluorophores which already existed, such as 1,8-naphthalimide, coumarin, rhodamine and their derivatives. Here, a new type of fluorescent molecule (BOTD) was synthesized and applied to the detection of H2S. The probe BOTD could quickly and sensitively detect H2S and turn on fluorescence. Moreover, the probe BOTD was successfully applied to the detection of exogenous and endogenous H2S in living cells, and may be expected to become a research tool for studying H2S-induced drugs.

A double-indole structure fluorescent probe for monitoring sulfur dioxide derivatives with distinct ratiometric fluorescence signals in mammalian cells.

Based on the addition reaction of the sulfur dioxide derivative to the CC double bond, the probe HDI was designed and synthesized. The two-channel fluorescent probe HDI changed from orange to colorless and the fluorescence changed from red to blue when the bisulfite was detected. And the probe responds rapidly to bisulfite within 2 min, with high sensitivity and specificity. In addition, the probe can be used to detect the concentration of bisulfite with a low detection limit (80 nM). Cytological experiments have also demonstrated that probe HDI has low cytotoxicity and could be used for ratiometric detection of sulfur dioxide derivatives in living cells.

[Randomized study of thalidomide combined with vinorelbine and cisplatin chemotherapy for the treatment of advanced non-small cell lung cancer].

OBJECTIVE: To evaluate the efficacy, median time to progression (TTP), quality of life and toxicity in the patients with advanced non-small cell lung cancer (NSCLC), treated with thalidomide plus vinorelbine and cisplatin (NP) or NP alone. METHODS: Sixty six patients with advanced NSCLC were divided randomly into two groups, the trial and control groups. The trial group was treated with vinorelbine 25 approximately 30 mg/m(2) i.v. on D1 and D8, cisplatin 70 approximately 80 mg/m(2) i.v. on D1 (NP regimen), and thalidomide 200 mg orally and daily from D1. The control group received vinorelbine and cisplatin as above described. RESULTS: Of 66 assessable patients, the overall response rate was 51.5% in the trial group and 36.4% in the control group (P = 0.22). The median TTP was 6.0 months for the trial group, and 3.6 months for the control group (P < 0.001). The score of quality of life in trial group was higher than that in the control group, but no significant difference was observed between the two groups (P > 0.05). There were no significant differences in toxicities between the two groups (P > 0.05). CONCLUSION: NP regimen combined with thalidomide can significantly prolong the median time to tumor progression in patients with advanced NSCLC. Thalidomide may have a synergic activity with NP regimen without increased toxicities.

[Analysis different transcriptional factors in different phenotype endometrial cancer cells].

OBJECTIVE: To analysis the activity of transcriptional factors in endometrial cancer cell lines with different estrogen receptor subtypes. METHODS: The mRNA levels of estrogen receptor (ER) was detected by quantitative RT-PCR, and the activity of transcriptional factors was also analysed by 345-channel protein/DNA array in RL-952 (the expression status of ERalpha and ERbeta both positive), HEC-1A [ERalpha (+/-), while ERbeta negative] and HEC-1B (ERalpha and ERbeta both negative). The transcription factors of NFkappaBp65 and p38MAPK with different activity were tested by enzyme-linked immunosorbent assay (ELISA) to confirm the results of protein/DNA array. RESULTS: The mRNA levels of ERalpha in RL-952, HEC-1A and HEC-1B were (6780+/-282), (684+/-84) and (168+/-38) copy/ng, respectively. Among 345 candidate transcriptional factors, there were 28 factors associated with ER status. Compared with RL-952 cells, 13 transcriptional activity factors were concomitantly up-regulation, while 15 concomitantly down-regulation in HEC-1A and HEC-1B cells. Transcriptional activities of TTF (1)-1, NRF-1, TCE were significantly correlated with the high-expression status of ERalpha mRNA (r=0.523, P=0.037), while RFX123 and Ikaros were significantly correlated with the low-expression status of ERalpha mRNA (r=-0.312, P=0.041). CONCLUSION: Transcriptional factors of TTF (1)-1, NRF-1, TCE may be associated with ER-mediated signal pathway, while RFX123 and Ikaros may be associated with non ER-mediated signal pathway in endometrial cancer.

Deciphering the Antitoxin-Regulated Bacterial Stress Response via Single-Cell Analysis.

Bacterial toxin-antitoxin (TA) systems, which are diverse and widespread among prokaryotes, are responsible for tolerance to drugs and environmental stresses. However, the low abundance of toxin and antitoxin proteins renders their quantitative measurement in single bacteria challenging. Employing a laboratory-built nano-flow cytometer (nFCM) to monitor a tetracysteine (TC)-tagged TA system labeled with the biarsenical dye FlAsH, we here report the development of a sensitive method that enables the detection of basal-level expression of antitoxin. Using the Escherichia coli MqsR/MqsA as a model TA system, we reveal for the first time that under its native promoter and in the absence of environmental stress, there exist two populations of bacteria with high or low levels of antitoxin MqsA. Under environmental stress, such as bile acid stress, heat shock, and amino acid starvation, the two populations of bacteria responded differently in terms of MqsA degradation and production. Subsequently, resumed production of MqsA after amino acid stress was observed for the first time. Taking advantage of the multiparameter capability of nFCM, bacterial growth rate and MqsA production were analyzed simultaneously. We found that under environmental stress, the response of bacterial growth was consistent with MqsA production but with an approximate 60 min lag. Overall, the results of the present study indicate that stochastic elevation of MqsA level facilitates bacterial survival, and the two populations with distinct phenotypes empower bacteria to deal with fluctuating environments. This analytical method will help researchers gain deeper insight into the heterogeneity and fundamental role of TA systems.

Potential role of the HOXD8 transcription factor in cisplatin resistance and tumour metastasis in advanced epithelial ovarian cancer.

Few studies have examined the potential transcription factor (TF) simultaneously associated with cisplatin resistance and metastasis in ovarian cancer. To assess a related mechanism, a 345-channel protein/DNA array and transcriptional activity ELISA were performed to compare the TF activities in the cisplatin-sensitive SKOV3 and cisplatin-resistant SKOV3-DDP cells and in HO-8910 and the homologous highly metastatic HO-8910PM cells. In SKOV3-DDP vs. SKOV3 cells, 43 TFs were up-regulated, while 31 were down-regulated. In HO-8910PM vs. HO-8910 cells, 13 TFs were up-regulated, while 18 were down-regulated. In these two models, 4 TFs (HOXD8(1), HOXD8(2), RB, RFX1/2/3) were simultaneously up-regulated, and 9 TFs (SRE, FKHR, Angiotensinogen ANG-IRE, Pax2, CD28RC/NF-IL2B, HLF, CPE, CBFB and c-Ets-1) were down-regulated. HOXD8 mRNA and protein expression levels measured by reverse transcription polymerase chain reaction and ELISA, respectively, were significantly higher in SKOV3-DDP and HO-8910PM than in their corresponding cell lines (both p < 0.05). In 52 cases of different ovarian disease, the patients with recurrent and cisplatin-resistant ovarian cancer had higher expression levels of HOXD8 than patients with primary malignant tumours (p = 0.018, p = 0.001) or benign tumours (p = 0.001, p < 0.001). Taken together, these results suggest that HOXD8 is potentially associated with both cisplatin resistance and metastasis in advanced ovarian cancer.

Regulation of matriptase and HAI-1 system, a novel therapeutic target in human endometrial cancer cells.

The effects of specific and non-specific regulation of matriptase on endometrial cancer cells in vitro were investigated. Messenger ribonucleic acid (mRNA) and protein expression of matriptase and hepatocyte growth factor activator inhibitor-1 (HAI-1) in RL-952, HEC-1A, and HEC-1B endometrial cancer cells were detected by real-time quantitative PCR (RT-qPCR) and western blot. The cells were infected with lentivirus-mediated small-interfering RNA (siRNA) targeted on matriptase (MA-siRNA) or treated with different cisplatin (DDP) concentrations. After treatment, invasion, migration, and cellular apoptosis were analyzed. Matriptase mRNA and protein expression significantly decreased to 80% after infection with MA-siRNA (P < 0.01), and scratch and trans-well chamber assays showed significant inhibition of invasiveness and metastasis. Upon incubation with cisplatin at concentrations higher than the therapeutic dose for 24 h, the expressions of matriptase and HAI-1 significantly decreased (P < 0.001). Moreover, the invasiveness, metastasis, and survival rate of HEC-1A and RL-952 endometrial cancer cells were significantly decreased (P < 0.001) due to the down-regulation of matriptase and HAI-1 upon increasing cisplatin concentration. However, a slight increase in matriptase and HAI-1 expression was observed in cells treated with low cisplatin concentration (P = 0.01). Moreover, matriptase expression was associated with metastasis and invasiveness. Down-regulation of matriptase by specific Ma-SiRNA or non-specific cisplatin in matriptase/HAI-1-positive endometrial cancer cells showed promising therapeutic features.

Cervical cancer screening using the Cervista high-risk human papillomavirus test: opportunistic screening of a hospital-based population in Fujian province, China.

OBJECTIVES: The Cervista® high-risk human papillomavirus (HR-HPV) test was evaluated as a primary screening method for cervical cancer in women aged ≥21 years and was compared with different screening and triage combinations. MATERIALS AND METHODS: A nested case-control study within the Fujian provincial Cervical Lesion Screening Cohorts was used to evaluate the Cervista test as the primary cervical screening method in a hospital-based population. Strategy 1 primarily screened using a cytology screen with HR-HPV testing used for triage. Strategy 2 primarily screened using cytology and HR-HPV co-testing. Strategy 3 primarily screened using HR-HPV testing and triaged HPV-positive women based on cytology. Strategy 4 primarily screened using HR-HPV testing and referred A9 pool HPV-positive women to colposcopy directly, whereas non-A9 HPV-positive women were triaged using cytology. RESULTS: There were 10,183 women included in this study; 16.49% (1677/10,183) were HR-HPV-positive, 9.52% had abnormal cytology, and 9907 women were normal during followup. A total of 276 women were diagnosed with cervical intraepithelial neoplasia 2 or worse (CIN2+), 197 with CIN3 or worse (CIN3+), and 70 with cervical cancer. Moreover, 10.15% (20/197) women who were CIN3+ were identified as cytology-negative, while 8.63% (17/197) were HR-HPV negative (P>0.05). The cumulative risk rate for HPV-/cytology- was 0.836 (95% CI, 0.424-1.648) in CIN3+ cases. Strategy 4 yielded the highest sensitivity for CIN2+ or CIN3+ and the lowest positive predictive value for CIN2+ or CIN3+ among the four screening strategies. CONCLUSION: The Cervista HR-HPV test can provide a reliable and sensitive clinical reference for the cervical cancer primary screen.