Effect of soil leaching on the toxicity thresholds (ECx) of Zn in soils with different properties.

Currently, the scientific basis for establishing soil environmental criteria is lacking. In order to establish reasonable soil environmental criteria values suitable for soils with different properties, this study selected soils from 16 different sites to determine the toxicity threshold of Zn based on toxicity tests of barley root elongation. In addition, leaching treatments were set up in seven soils with different properties to eliminate the influence of the accompanying anions (Cl-) on the determination of the Zn toxicity threshold. The results indicated that the toxicity thresholds of different soils vary greatly. The EC10 and EC50 ranges of barley root elongation in 16 kinds of non-leached soils were 18.5 mgkg-1 to 1618.7 mgkg-1 and 277.9 mgkg-1 to 3179.8 mgkg-1, respectively. The hormesis effect appeared in the dose response of Zn, and relative barley root elongation reached more than 150%. Leaching significantly reduced the Zn toxicity in acidic soils. The variation ranges of the leaching factor (LF) in the seven soils were LF10 = 1.1-9.3, LF50 = 1.0-3.2. The LF prediction model indicated that pH explained 81.4% of the LF variation (p < 0.01). The soil pH, cation exchange capacity (CEC), and conductivity (EC) explained 97.8% of the EC50 variation in the leached soil (p < 0.01). The results provide reference values for Zn environmental criteria.

Sulfur application modifies cadmium availability and transfer in the soil-rice system under unstable pe+pH conditions.

The objective of this study was to investigate the responses of cadmium (Cd) availability and transfer in the soil-rice system to added sulfur (S) under unstable pe + pH conditions. Different water management conditions (flooding and aerobic treatments) cause changes in the soil pe + pH. We conducted a pot experiment to investigate the influence of S supply on soil Cd availability and Cd accumulation in rice plants (Oryza sativa L.), using three water regimes (continuous dryness, alternating dry-wet for one cycle, and continuous flooding) combined with two S concentrations (0 and 300 mg/kg). The results showed that the flooding treatment was more effective in decreasing soil pe + pH, Cd availability, and Cd accumulation in rice tissues than were the aerobic treatments. S-induced reduction in Cd uptake and translocation in rice was attributed to the decreased soil pe + pH values and enhanced biosynthesis of phytochelatins (PCs) and glutathione (GSH) in rice roots. Microscopic examination showed that the flooding treatment with added soil S resulted in better rice root growth. Element dispersive spectrometer (EDS) analysis indicated that S addition and flooding treatment promoted the formation of iron plaques and increases in Fe concentration in rice tissues. Conversely, partial disintegration of the root epidermis was observed in the dry treatment without added S.

Total saponins of Bolbostemma paniculatum (maxim.) Franquet exert antitumor activity against MDA-MB-231 human breast cancer cells via inhibiting PI3K/Akt/mTOR pathway.

BACKGROUND: The aim of the present study was to examine the effects of the Bolbostemma paniculatum (Maxim.) Franquet (BP) active compound, BP total saponins (BPTS), on MDA-MB-231 cells, and investigate the underlying mechanism regarding BPTS-mediated attenuation of the PI3K/Akt/mTOR pathway. METHODS: The effect of BPTS on cytotoxicity, induction of apoptosis and migration on MDA-MB-231 cells at three different concentrations was investigated. A CCK-8 assay, wound-healing assay and flow cytometry were used to demonstrate the effects of BPTS. Additionally, expression of the primary members of the PI3K/Akt/mTOR signaling pathway was assessed using western blotting. To verify the underlying mechanisms, a PI3K inhibitor and an mTOR inhibitor were used. RESULTS: BPTS inhibited proliferation of MDA-MB-231 cells with an IC50 value of 10 µg/mL at 48 h. BPTS inhibited migration of MDA-MB-231 cells, and the western blot results demonstrated that BPTS reduced p-PI3K, p-Akt and p-mTOR protein expression levels in MDA-MB-231 cells. Additionally, the results were confirmed using a PI3K inhibitor and an mTOR inhibitor. BPTS decreased proliferation and migration of MDA-MB-231 cells possibly through inhibiting the PI3K/Akt/mTOR signaling pathway. CONCLUSIONS: The results highlight the therapeutic potential of BPTS for treating patients with triple-negative breast cancer.

Clinical evaluation of human papillomavirus 16/18 oncoprotein test for cervical cancer screening and HPV positive women triage.

HPV-16 and -18 account for about 80% of cervical cancers. We evaluated the performance of HPV-16/18 oncoprotein to predict precancer and cancer in corresponding tissue biopsy specimens. 1,008 women attending cervical cancer screening program and 638 women referred to colposcopy with biopsy-confirmed cervical intraepithelial neoplasia grade 2 or worse (CIN2+) from 4 hospitals were recruited (1,646 in total). All women were tested OncoE6 (AVC), Liquid-Based Cytology (Hologic) and cobas HPV test (Roche). Colposcopy was performed on women with any abnormal results. The final diagnoses were based on a consensus panel review of the histology. There were 919 normal, 69 CIN1, 53 CIN2, 91 CIN3,474 squamous cell carcinoma(SCC) and 40 adenocarcinoma (ADC) cases, the prevalence of OncoE6 was 1.7%, 10.1%, 13.2%, 44.0%, 80.4% and 65.0%, respectively. The percent positive for cobas was higher than that of OncoE6 in detection of HPV16/18 in entire population (p < 0.001). However, the disparity of positive rate between these two tests became tiny among cervical cancer patients (CIN2: 26.4% vs. 13.2%, CIN3: 73.6% vs. 44.0%, SCC: 84.0% vs. 80.4%, ADC: 67.5% vs. 65.0%). OncoE6 was less sensitive than cobas (73.9% vs. 93.6%, p < 0.001), but more specific (97.1% vs. 75.4%, p < 0.001) for CIN3+ in entire population; OncoE6 yielded a sensitivity of 77.7% and a specificity of 91.0% for CIN3+ among cobas positive women, which can reduce nearly half of the colposcopy referral numbers. OncoE6 can be considered as a useful tool for cervical cancer screening and a potential powerful biomarker for HPV positive triage.

[Association between life style, diet intake and high risk-human papillomavirus persistent infection among rural women in Xinmi City, Henan Province].

OBJECTIVE: To explore the association between life style, diet intake and high risk human papillomavirus (HR-HPV) persistent infection among Chinese rural women living in Xinmi City, Henan Province. METHODS: In 2010, a 3-year prospective study in which 2500 women were enrolled and screened by different HR-HPV DNA tests was conducted, part of women among them was followed and tested for HR-HPV DNA in 2012 and 2014. Furthermore, socio demographic factors, gynecological information and diet intake in the past 12 months were collected by self-designed questionnaire in 2014. A total of 721 women with complete test results were eligible for the final analysis. Study participants were divided into 3 groups (persistent infection group, transient infection group, and negative group) by HR-HPV status, and the association between life style, diet intake and HR-HPV persistence was evaluated using ordinal logistic regression model. RESULTS: The average age of 721 women included in the analysis was 50 years old. 141 women had HR-HPV persistent infection, 180 women had HR-HPV transient infection, and 400 women were negative for HR-HPV in 3 years. Age (Χ2 = 58.449, P < 0.001, P(trend) < 0.001), smoking (Χ2 = 6.981, P = 0.021), contraception method (Χ2 = 8.448, P = 0.015) , menopause (Χ2 = 35.712, P < 0.001), number of live births (Χ2 = 16.340, P < 0.001, P(trend) < 0.001), and the intake frequency of cereals (Χ2 = 17.937, P = 0.001) or others (Χ2 = 12.107, P = 0.017) varied significantly between women grouped by different HR-HPV status. Compared to women who were older than 59 years, women in the younger groups had a much lower risk of HR-HPV persistence (adjusted OR1 = 0.39, 95% CI 0.26 - 0.59, OR2 = 0.40, 95% CI 0.23 - 0.69, and OR3 = 0.28, 95% CI 0.12 - 0.68). CONCLUSION: Age is the main risk factor of HR-HPV persistent infection. Lifestyle, diet intake do not associate with HR-HPV persistence after adjustment by age.

[Regulatory effects of ninjurin-1 on adhesion of myeloid cells in the retina at the early stage of diabetic rats].

OBJECTIVE: To investigate the regulatory effects of ninjurin-1 on adhesion of myeloid cells in the retina at the early stage of diabetic rats. METHODS: Experimental study. The rat diabetic model was induced by intraperitoneal injection of streptozotocin. After 2 months of diabetes induction, 27 diabetic rats were randomly chosen and assigned to 3 groups, including diabetes and phosphate buffered saline (PBS) injection group (group B), diabetes and anti-Ninj-1 injection group (group C) and diabetes and anti-IgG injection group (group D), with 9 rats in each group. Nine age matched health rats were chosen as control group (group A). Retinal leukostasis was quantified with acridine orange leukocyte fluorography. Retinal myeloid cell infiltration activity was measured by enzyme linked immunosorbent assay of myeloperoxidase (MPO). The differences of the mean values among the four groups were analyzed by one-factor analysis of variance. The multiple comparisons of the mean values among the four groups were analyzed by LSD-t analysis. RESULTS: According to the results of the acridine orange leukocyte fluorography, the numbers of leukocyte adhesion in the four groups were 49.66 ± 13.51, 153.66 ± 20.43, 85.33 ± 15.03 and 156.33 ± 11.53, respectively. The differences among them were significant (F = 143.34, P = 0.000). The numbers of leukocyte adhesion in the group C were significantly lower than that in group B (P = 0.000, 95%CI: -82.68 - -53.98). The levels of retinal MPO in the four groups were (15.66 ± 2.08), (27.66 ± 2.51), (18.02 ± 2.01) and (26.66 ± 3.21) µg/L, respectively. The differences among them were significant (F = 17.61, P = 0.010). The level of retinal MPO in the group C was significantly lower than that in group B (P = 0.010, 95%CI: -14.37 - -4.95). CONCLUSIONS: Ninj-1 may play a role in the mediation of the adhesion of myeloid cell to the rat retina of early-stage of diabetes in vivo.

Astragalus mongholicus Bunge and Panax notoginseng formula (A&P) improves renal mesangial cell damage in diabetic nephropathy by inhibiting the inflammatory response of infiltrated macrophages.

BACKGROUND: Diabetic nephropathy (DN) is one of the main causes of end-stage renal disease with scantly effective treatment. Numerous evidences indicated that macrophages play an important role in the occurrence and pathogenesis of DN by secreting inflammatory cytokines. Mincle is mainly expressed in macrophages and promotes kidney inflammation and damage of acute kidney injury. However, the role of Mincle in DN is unclear. In this study, we aim to investigate the effect of Mincle-related macrophage inflammation on DN, and whether it can be identified as the therapeutic target for Astragalus mongholicus Bunge and Panax notoginseng Formula (A&P), a widely used Chinese herbal decoction for DN treatment. METHODS: In vivo experiments high-fat and high-sugar diet and streptozotocin was used to establish a diabetic nephropathy model, while in vitro experiments inflammation model was induced by high-glucose in mouse Bone Marrow-Derived Macrophages (BMDM) cells and mouse mesangial (MES) cells. Kidney pathological staining is used to detect kidney tissue damage and inflammation, Western blotting, Real-time PCR and ELISA are performed to detect Mincle signaling pathway related proteins and inflammatory cytokines. RESULTS: Mincle was mainly expressed in infiltrated macrophage of DN kidney, and was significant decreased after A&P administration. The in vitro experiments also proved that A&P effectively down-regulated the expression of Mincle in macrophage stimulated by high glucose. Meanwhile, the data demonstrated that A&P can reduce the activation of NFκB, and the expression and secretion of inflammatory cytokines in DN kidney or BMDM cells. Notably, we set up a co-culture system to conform that BMDM cells can aggravate the inflammatory response of mesangial (MES) cells under high glucose stimulation. Furthermore, we found that the anti-injury role of A&P in MES cells was dependent on inhibition of the Mincle in macrophage. CONCLUSION: In summary, our study found that A&P is effective in reducing renal pathological damage and improving renal function and inflammation in diabetic nephropathy by a mechanism mainly related to the inhibition of the Mincle/Card9/NFκB signaling pathway.

Cyclophosphamide inhibits Pax5 methylation to regulate the growth of retinoblastoma via the Notch1 pathway.

Retinoblastoma (Rb) is the most common intraocular malignant tumor in infants. Here, we investigated the function and mechanism of cyclophosphamide (CTX) in the development of Rb. Real-time quantitative polymerase chain reaction (RT-qPCR) results showed that paired box protein 5 (Pax5) expression was down-regulated in Rb tissues and cell lines. Methylation-specific PCR (MSP) results showed that the methylation level of Pax5 was up-regulated in Rb. After treatment with CTX, the Pax5 expression in Rb cell lines was increased significantly. The methylation of Pax5 and the expression of DNA methyltransferases (DNMTs) were down-regulated in the CTX group. Cyclophosphamide inhibited cell proliferation, migration, and invasion, promoted cell apoptosis via the Notch1 pathway. DNA methyltransferase inhibitor SGI-1027 had synergistic effects with CTX. Paired box protein 5 siRNA was transfected into Y79 cells treated with CTX. The expression of DNMTs, Pax5, the Notch1 pathway and apoptosis marker protein was detected by Western blotting, and changes in cell behavior were detected, respectively. Results showed that knockdown of Pax5 reversed the effects of CTX. Moreover, the Notch1 activator Valproic acid (VPA) abolished the inhibitory effects of CTX on Rb development. Moreover, CTX inhibited tumor growth in nude mice. These findings demonstrated that CTX up-regulated Pax5 expression by down-regulating DNMTs expression, and then inhibited the Notch1 signaling pathway activation and Rb growth.

Redistribution of iron oxides in aggregates induced by pe + pH variation alters Cd availability in paddy soils.

In this study, the effect of unstable pe + pH levels on the transformation of Fe oxides in different-sized soil fractions and its impact on Cd speciation were explored. Paddy soil samples collected from two locations in China were cultivated for two months under one of four pe + pH conditions: flooding + N2 (T1), flooding (T2), 70% water holding capacity (T3), and 70% water holding capacity + O2 (T4). Chemical analysis and X-ray diffraction (XRD) were used to identify the mineralogical phases and species of Fe and Cd in paddy soils. The results show that the decrease of soil pe + pH level favored the transformation of well-crystallized Fe oxides (Fec), such as hematite and goethite, into poorly-crystallized (Feo) and organically-complexed (Fep) forms. The transformation promoted the binding of Cd to Fe oxides and was primarily responsible for up to a 41.8% decrease of soil DTPA (diethylenetriaminepentaacetic acid)-extractable-Cd content. In addition, the decline in pe + pH value reduced Fe concentrations in soil particle fractions of 0.2-2-mm (17.8%-30.6%) and <0.002-mm (20.7%-31.7%) of the two flooding treatments. The decreased Fe concentrations were closely associated with less Fec contents in these same fractions and more Feo and Fep in coarser aggregates (P < 0.01). Importantly, the increase in contents of Feo and Fep in the 0.002-2 mm fraction were significantly correlated with content of Fe-/Mn-oxide-bound Cd (OX-Cd) in larger particle-size fractions (P < 0.01). Furthermore, the increasing content of OX-Cd played a crucial role in reducing DTPA-Cd content. This study demonstrates that low pe + pH values favor the transformation of crystalline Fe oxides into a poorly-crystallized and organically-complexed phase, which facilitates Cd accumulation in coarser aggregates and enhances Cd stability in paddy soils.

Fe(III) reduction due to low pe+pH contributes to reducing Cd transfer within a soil-rice system.

Effect of Fe redox state caused by low soil pe+pH levels on Cd uptake by rice is unclear. Rice grown in pots of Cd-contaminated paddy soil were subjected to different irrigation regimes: flooding, intermittent flooding (Int-FL), and sustained soil moisture at 70% water holding capacity (WHC). Results showed low pe+pH (5.52 and 7.09) in flooding treatment significantly increased relative abundances of Fe-reducing bacteria (FeRB) (6.29% and 4.51%), especially members within the Clostridium, Geobacter and Desulfuromonadia genera. Stimulation of FeRB activity induced Fe(III) reduction and increased Fe2+ content in flooded soils, which promoted Cd sequestration in low-crystalline fraction of IP (IP-Feh-Cd) and Cd bonded to amorphous Fe-oxides (amFeox-Cd). The 24.9-62.4% higher amFeox-Cd content was the important factor for 20.4-44.2% lower CaCl2-extractable Cd content in flooding treatment than those in other treatments. Soil submergence reduced Cd uptake by rice at tillering and booting stages, the critical periods of Cd transport in the soil-rice system, which was attributed to the increases in dissolved Fe2+ and IP-Feh-Cd contents and decrease in CaCl2-Cd content. Therefore, maintaining flooding during the tillering and booting stages may be an effective strategy to reduce Cd uptake by rice cultivated in Cd-contaminated soil.

Manganese facilitates cadmium stabilization through physicochemical dynamics and amino acid accumulation in rice rhizosphere under flood-associated low pe+pH.

Periodic flooding in paddy soils impacts redox behavior and induces variations in pe+pH levels. Manganese (Mn) is capable of reducing cadmium (Cd) uptake by rice. However, the processes involved in how Mn alters Cd mobilization under different pe+pH environments remain poorly understood. To investigate the mechanisms of Mn-mediated soil Cd-stabilization and subsequent inhibition of Cd uptake from flooded soils, we examined Cd immobilization in soil pot incubations, transcriptional changes in Cd-transport genes, and metabolomic analyses of roots and rhizosphere soils with or without Mn application. We found a decrease in extractable Cd concentration largely depended on irrigation-associated low pe+pH, exogenous Mn enhancement of Fe-Mn (oxyhydro)oxide-mediated Cd transformation, and Cd deposition in rice Fe/Mn plaques. Mn application led to striking effects on the expression of Cd-related genes eg. IRT, HMA, and NRAMP in rice root tissue. Exposure to Mn under variable pe+pH levels resulted in metabolic reprogramming of soil and rice roots. Mn induced amino acid synthesis in rice roots, leading to rhizosphere accumulation of free L-lysine, glycine, and glutamine, which can reportedly bind metal ions, forming complexes with Cd. Thus, secreted amino acids, low pe+pH, and free Mn can together comprise a multi-faceted approach to managing Cd toxicity in rice.

MicroRNA­363­3p inhibits cell proliferation and induces apoptosis in retinoblastoma cells via the Akt/mTOR signaling pathway by targeting PIK3CA.

There is extensive evidence suggesting that microRNAs (miRs) can modulate the activity of oncogenes and tumor suppressors, and are associated with the occurrence of cancer. In the present study, the function of miR­363­3p in the progression of retinoblastoma (RB) was investigated. miR­363­3p expression in RB was decreased, and miR­363­3p protein levels were found to be inversely correlated with phosphatidylinositol­4,5­bisphosphate 3­kinase catalytic subunit α (PIK3CA) levels. Overexpression of miR­363­3p in an in vitro model of RB revealed that miR­363­3p had anticancer effects on RB and regulated PIK3CA, pyruvate dehydrogenase kinase 1 (PDK1) and phosphorylated protein kinase B (p­AKT) protein expression. Downregulation of miR­363­3p promoted cell proliferation of RB cells through PIK3CA, PDK1 and p­AKT protein expression. Knockdown of PIK3CA increased the anticancer effects of miR­363­3p in RB cells. Treatment with OSU­03012, a PDK1 inhibitor, accelerated the anticancer effects of miR­363­3p in RB cells. Taken together, the results demonstrate that miR­363­3p functions as a tumor suppressor in RB by targeting PIK3CA.

Iron fractions responsible for the variation of Cd bioavailability in paddy soil under variable pe+pH conditions.

Iron (Fe) in soil is closely related to cadmium (Cd) uptake by rice plants, and soil pe + pH significantly influences Fe redox behavior. This study aimed to explore the influential mechanisms of varying pe + pH conditions on the transformation of iron oxides in the rhizosphere and the subsequent effect on Cd accumulation in rice plants. A two-month pot experiment was conducted to investigate the effect of soil pe + pH on the fractions of iron oxides and formation of iron plaque (IP), as well as the effect of these changes on Cd uptake by rice plants (Oryza sativa L.). Different irrigation strategies, 70% water holding capacity (DY), continuous flooding (FL), and alternate flooding/drying weekly (AWD), were used to achieve various soil pe + pH levels. The results showed that low pe + pH conditions (under the FL and AWD treatments) were more beneficial to the transformation of crystalline iron oxides into amorphous forms in rhizosphere soil and the precipitation of IP on rice roots. The increase of amorphous iron oxides resulted in the reduction of Cd availability in rhizosphere soil by immobilizing more Cd on Fe oxides. Moreover, Cd adsorbed on rice root surfaces reacted with IP, inhibiting Cd soil-to-root transport. The two mechanisms combinatively functioned at decreasing Cd concentration in rice shoots by 14.1-33.1% at low pe + pH conditions compared to that of the high pe + pH (DY treatment). These results indicate that lowering soil pe + pH effectively reduced Cd accumulation in rice plants, probably through the immobilization of amorphous Fe oxides on Cd and sequestration of iron-plaque on Cd.

Protective effects of the Chinese herbal medicine prescription Zhujing pill on retina of streptozotocin-induced diabetic rats.

OBJECTIVE: In this study, we investigate the protective effects of the Chinese herbal medicine prescription Zhujing pill (ZJP) on the development of diabetic retinopathy (DR) and explore the potential mechanisms. MATERIALS AND METHODS: Zhujing pill extract (ZJPE) was prepared, and the main components were identified by high-performance liquid chromatography (HPLC). Several serum and tissue (retina) parameters were measured, such as levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), intercellular adhesion molecule 1 (ICAM-1), vascular endothelial growth factor (VEGF), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), advanced glycation end products (AGEs), and high sensitivity C-reactive protein (hsCRP). Aldose reductase (AR) activity and blood-retinal barrier (BRB) breakdown were determined. Finally, retinal electrophysiology and morphological changes were assayed. RESULTS: In STZ-induced DM rats, ZJPE treatment restored the body weight decrease. DM rats showed decreased levels of SOD and GSH-Px and increased levels of IL-6, TNF-α, hsCRP, and MDA, whereas all these changes were significantly reversed by ZJPE administration. ZJPE alleviated BRB breakdown. Furthermore, ZJPE also alleviated the retinal electrophysiology changes and impaired morphology of the retina and lowered the high levels of TNF-α, IL-1ß, ICAM-1, VEGF, AGEs, and AR in the retina. CONCLUSIONS: ZJPE treatment attenuated the progression of DR in STZ-induced diabetic rats.

Typhae pollen polysaccharides ameliorate diabetic retinal injury in a streptozotocin-induced diabetic rat model.

ETHNOPHARMACOLOGICAL RELEVANCE: According to ancient traditional Chinese medicine, Typhae Pollen (TP) is commonly used to treat fundus haemorrhage because it improves blood circulation. AIMS OF THE STUDY: This study evaluated the role of the main TP component, polysaccharides (TPP), on diabetic retinopathy (DR) and its possible mechanisms of inhibiting inflammation and improving blood circulation. MATERIALS AND METHODS: After successful establishment of a diabetic rat model, TPP was administered to diabetic rats for treatment, and the rats were sacrificed at 12 weeks. Retinal electrophysiology and ultrastructures were observed, and serum interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) levels were also measured. Changes in the retinal expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were examined by immunofluorescence. A mouse model of acute blood stasis was then established, and the effects of TPP on haemorheology were observed. The anti-inflammatory effect of TPP was analysed based on the changes in abdominal capillary permeability and the degree of auricle swelling in the mice. RESULTS: In streptozotocin (STZ)-induced DR rats, TPP (0.4 g/kg) treatment restored electrophysiology indexes and retinal ultrastructures, reduced serum IL-6 and TNF-α levels, decreased VEGF and bFGF expression in retinal tissues, and improved haemorheology indexes. Moreover, TPP reduced abdominal capillary permeability and relieved auricle swelling in a dose-dependent manner. CONCLUSIONS: TPP treatment ameliorated DR by inhibiting inflammation and improving blood circulation.

The association between human papillomavirus 16, 18 DNA load and E6 protein expression in cervical intraepithelial neoplasia and cancer.

BACKGROUND: It has been reported that high HPV DNA load and elevated E6 protein expression correlate with cervical cancer, but no epidemiological study has been performed. OBJECTIVES: To assess the association between type-specific HPV DNA load and presence of E6 protein in cervical intraepithelial neoplasia and cancer. STUDY DESIGN: This study was a cross-sectional multicenter study performed between 2013 and 2017. A total of 1717 women (normal histopathology: n = 916; CIN1: n = 94; CIN2: n = 63; CIN3: n = 130; SCC: n = 474; adenocarcinoma: n = 40) were included. HPV DNA load and presence of E6 protein were detected. DNA load was measured as log copies/10,000 cells. RESULTS: The HPV16/18-E6 positivity rates increased from negative for DNA to the highest load grade. Compared to the HPV16 DNA negatives, women with low (RR = 31.5, 95%CI = 18.9-52.5), medium-low (RR = 133.5, 95%CI = 77.3-230.7), medium-high (RR = 247.9, 95%CI = 134.9-455.6) and high DNA loads (RR = 677.9, 95%CI = 301.6-1523.7) had increasingly higher relative ratios of HPV16-E6 expression. The association of HPV18-E6 with its DNA load was also significant for low (RR = 27.9, 95%CI = 10.4-74.9), medium-low (RR = 89.0, 95%CI = 32.8-241.3), medium-high (RR = 276.8, 95%CI = 76.7-998.9) and high grade (RR = 441.2, 95%CI = 97.7-1992.4). The positivity rates of both HPV16 DNA and E6 protein increased consistently with the severity of diseases from normal histopathology to SCC. Unlike HPV16, the trends of HPV18 DNA and E6 protein fluctuated consistently among women from normal histopathology to cancer. DNA load in E6-positive women was significantly higher than E6-negative for both types. CONCLUSIONS: There is a type-dependent association between HPV16/18 DNA load and E6 protein; our study furthers the understanding of the natural history of cervical cancer.

MicroRNA-486-5p suppresses TGF-ß2-induced proliferation, invasion and epithelial-mesenchymal transition of lens epithelial cells by targeting Smad2.

The pathological development of lens epithelial cells (LECs) leads to posterior capsular opacification (PCO). This study was undertaken to investigate the effects of microRNA-486-5p (miR-486-5p) on TGF-ß2-induced proliferation, invasion and epithelial-mesenchymal transition (EMT) in the lens epithelial cell line SRA01/04, and to explore the underlying molecular mechanisms. The expression of miR-486-5p in TGF-ß2-induced SRA01/04 cells was down-regulated, and the expression of Smad2, p-Smad2 and p-Smad3 was up-regulated. A dual-luciferase reporter assay revealed that miR-486-5p directly targets the 30'-UTR of Smad2. MiR-486-5p mimic transfection markedly down-regulated the expression levels of Smad2, thus inhibiting the expression of p-Smad2 and p-Smad3. MiR-486-5p overexpression in SRA01/04 cells markedly suppressed TGF-ß2-induced proliferation and invasion, inhibited protein expression of CDK2 and CDK4, down-regulated fibronectin, α-SMA and vimentin and up-regulated E-cadherin; these effects were partly reversed by Smad2 overexpression. In short, these data show that miR-486-5p overexpression can inhibit TGF-ß2-induced proliferation, invasion and EMT in SRA01/04 cells by repressing Smad2/Smad3 signalling, implying that miR-486-5p may be an effective target to interfere in the progression of PCO.

p16/Ki-67 co-expression associates high risk human papillomavirus persistence and cervical histopathology: a 3-year cohort study in China.

PURPOSE: To evaluate the association of p16/Ki-67 co-expression and persistence of high-risk human papillomavirus (HR-HPV) infection as well as cervical abnormalities. METHODS: We performed a 3-year cohort study among which 2498 Chinese women aged 25 to 65 years were screened by different HPV tests in 2011. 690 women who were positive at any of the tests and a random sample of 164 women with all negative results received colposcopy, cervical specimens for cobas HPV test (Roche diagnostics) were collected before colposcopy; of this group, 737 cervical specimens were collected to perform cobas, Liquid-based cytology, HPV E6 test (Arbor Vita Corporation) and p16/Ki-67 dual staining (Roche diagnostics) in 2014. Colposcopy and biopsies was performed on women with any abnormal result. RESULTS: Compared to women without HR-HPV persistent infection, women in the HR-HPV persistence group had a higher risk of p16/Ki-67 positive, with an adjusted Odds Ratio(OR) and 95% confidence interval (CI) of 6.29 (4.07-9.72); moreover, adjusted odds ratio for women who had HPV16/18 persistent infection was nearly 4-folder higher than women with other 12 HR-HPV persistent infection (adjusted OR = 17.15, 95% CI: 7.11-41.33 vs adjusted OR = 4.68, 95% CI: 2.89-7.58). Additionally, p16/Ki-67 positivity rate significantly increased with the severity of the cytological and histological abnormalities, and resulted strongly associated with a CIN2+ diagnosis (OR = 16.03, 95% CI: 4.46-57.59). CONCLUSIONS: p16/Ki-67 co-expressions associated strongly with HR-HPV persistence, especially with HPV16/18, and the presence of a CIN2+ lesion. Therefore, p16/Ki-67 could be considered as a suitable biomarker for cervical cancer screening, particularly in HPV-based screening programs.

Evaluation of human papillomavirus detection by Abbott m2000 system on samples collected by FTA Elute™ Card in a Chinese HIV-1 positive population.

BACKGROUND: HIV+/AIDS women have an increased risk of developing into CIN and cervical cancer compared to the general population. Limited medical resource and the lack of AIDS relevant knowledge impair the coverage and efficiency of cervical cancer screening. OBJECTIVES: To compare the clinical performance of self-collected dry storage medium (FTA Elute card) and physician-collected PreservCyt medium in detection of high risk human papillomavirus (HR HPV) among HIV-1 positive population. METHODS: Three hundred HIV-1 positive women (aged 25-65) were recruited from Yunnan infectious hospital. Two cervicovaginal samples were collected from each participant: one was collected by the women themselves and applied on a FTA Elute card; the other one was collected by a physician and stored in PreservCyt solution. All the samples were tested for 14 HR HPV using Abbott RealTime High Risk HPV assay. Biopsies were taken for histological diagnosis if any abnormal impression was noticed under colposcopy. RESULTS: 291 (97.0%) of participants were eligible for this study. 101 (34.70%) participants were found HR HPV positive in both FTA card and PreservCyt samples, and 19 (6.53%) women were diagnosed as CIN2+. The HR HPV positive rate on samples collected by FTA Elute card and PreservCyt solution was 42.61% and 39.86%, respectively. The overall agreement was 87% (kappa=0.731) between FTA card and PreservCyt. The clinical sensitivity and specificity of FTA card and PreservCyt were 100%, 61.39% and 100%, 64.33%, respectively. CONCLUSIONS: In this study, FTA Elute card demonstrated a good performance on self-collected sample for HR HPV detection in HIV-1 positive population. For the women from low-resource area with HIV-1 infection, FTA Elute card could be an attractive sample collection method for cervical cancer screening.

Elevated Expression of Human Papillomavirus-16/18 E6 Oncoprotein Associates with Persistence of Viral Infection: A 3-Year Prospective Study in China.

BACKGROUND: An association between high-risk human papillomavirus (HR-HPV) oncoprotein expression and viral persistence has been suggested by the outcome of etiology studies, but there are no epidemiologic studies evaluating that link. METHODS: We performed a 3-year prospective study in which 2,498 Chinese women ages 25 to 65 years were screened by six screening tests, including the Onco E6: Cervical Test (Arbor Vita Corporation) in 2011 (baseline). Six-hundred and ninety women who were positive for any of the tests and a random sample of 164 women with all negative results received colposcopy, and cervical specimens for the cobas 4800 HPV test ("cobas," Roche Molecular Systems) were collected before colposcopy; of this group, 737 cervical specimens were collected to perform cobas and Onco E6: Cervical Test in 2014 (follow-up). Twenty-four cases of HPV16/18 E6 positives and 204 selected controls at baseline, 13 cases of HPV16/18 E6 positive and another 204 selected controls at follow-up were analyzed separately using unconditional logistical regression models to estimate ORs and 95% confidence intervals (CI). RESULTS: Compared with women who were HPV16 E6 oncoprotein negative at baseline, women in the E6-positive group had a much higher risk of HPV persistence (adjusted OR, 54.64; 95% CI, 7.19-415.09) at 3-year follow-up; a statistically strong association was also found between HPV16/18 HPV persistence and E6 oncoprotein expression detected at follow-up (adjusted OR, 360.57; 95% CI, 28.30-4,593.55). CONCLUSIONS: A single detection of HPV16/18 E6 oncoprotein expression was strongly associated with viral persistence. IMPACT: HPV16/18 E6 oncoprotein constitutes a marker for risk of HPV persistence. Cancer Epidemiol Biomarkers Prev; 25(7); 1167-74. ©2016 AACR.