M2 macrophage-derived exosomal circTMCO3 acts through miR-515-5p and ITGA8 to enhance malignancy in ovarian cancer.

Tumor-associated macrophages of the M2 phenotype promote cancer initiation and progression. Importantly, M2 macrophage-derived exosomes play key roles in the malignancy of cancer cells. Here, we report that circTMCO3 is upregulated in ovarian cancer patients, and its high expression indicates poor survival. M2-derived exosomes promote proliferation, migration, and invasion in ovarian cancer, but these effects are abolished by knockdown of circTMCO3. Furthermore, circTMCO3 functions as a competing endogenous RNA for miR-515-5p to reduce its abundance, thus upregulating ITGA8 in ovarian cancer. miR-515-5p inhibits ovarian cancer malignancy via directly downregulating ITGA8. The decreased oncogenic activity of circTMCO3-silencing exosomes is reversed by miR-515-5p knockdown or ITGA8 overexpression. Exosomal circTMCO3 promotes ovarian cancer progression in nude mice. Thus, M2 macrophage-derived exosomes promote malignancy by delivering circTMCO3 and targeting the miR-515-5p/ITGA8 axis in ovarian cancer. Our findings not only provide mechanistic insights into ovarian cancer progression, but also suggest potential therapeutic targets.

Adsorption properties and mechanism research of phosphorus with different molecular structures from aqueous solutions by La-modified biochar.

In order to explore the adsorption characteristics of phosphorus from molecules with different molecular structures and varying number of phosphate groups on metal-modified biochar, walnut shell biochar was modified with LaCl3 to prepare lanthanum-loaded biochar (BC-La). Adsorption of four polar components, namely phytic acid (IHP), adenosine-5'-disodium triphosphate (5-ATP), hydroxyethylidene diphosphonic acid (HEDP), and sodium pyrophosphate (PP), was studied. The adsorption properties and mechanism of phosphorus sorption by BC-La were analyzed by SEM-EDS and FTIR for the different structures. The results showed that the maximum adsorption capacity of BC-La for IHP, 5-ATP, HEDP, and PP was 85.85, 9.04, 15.80, and 14.45 mg/g, respectively. The adsorption capacity was positively correlated with the polarity of organic phosphorus. The adsorption behavior conformed to the quasi second-order kinetic fitting equation, and the increase of temperature was conducive to the removal of all four phosphorus pollutants. BC-La adsorbs IHP and HEDP mainly through electrostatic attraction. The adsorption of 5-ATP and PP is dominated by complexation. The La-modified biochar has broad prospects in water remediation, which can provide a theoretical basis for removal of different forms of phosphorus pollutants and prevention and control of water eutrophication.

Modified biochar improves the storage capacity and adsorption affinity of organic phosphorus in soil.

The loss of soil organic phosphorus can easily cause water eutrophication. In order to effectively reduce the loss of soil organic phosphorus, this manuscript investigated the adsorption of soil organic phosphorus by lanthanum modified biochar (BC), traditional adsorbent gypsum (GY) and zeolite (ZE) by taking phytic acid as the representative. The adsorption isotherm model and kinetic models were used to fit the phosphorus absorption characteristics of the adsorbents. The effects of initial pH and temperature on the adsorption capacity were discussed, and the adsorption mechanism of each adsorbent was explained by means of FTIR and XRD. The results showed that the adsorption capacity of phytate phosphorus followed the trend of BCTS > GYTS > ZETS > TS (soil), and the maximum phosphorus adsorption capacity obtained from Langmuir isotherm for treatment with BCTS was 2.836 mg g-1, and the treatment had the strongest affinity for phytate phosphorus and also the ability to store phosphorus. The adsorption process fits well with Langmuir isotherm equation and pseudo-second-order kinetic equation, and the adsorption behavior of phytate phosphorus was mainly controlled by the chemisorption of monolayer. When the concentration of phytate phosphorus was 100 mg L-1, percentage of modified biochar added to the soil was 3% and the pH was 6, the adsorption capacity reached the maximum, and the maximum adsorption capacity was 2.000 mg g-1. The results of FTIR and XRD characterization showed that complexation was the main adsorption mechanism. In this study, the combination of modified biochar and soil phytate phosphorus can provide a good theoretical basis for reducing the loss of soil organic phosphorus.

[Effects of Different Fertilization Patterns on Nitrogen Leaching Loss from Paddy Fields Under Reduced Nitrogen].

Nitrogen leaching loss in paddy fields is one of the main ways of farmland non-point source pollution. To explore the suitable fertilization of rice fields in the Erhai Lake Basin and reduce the nitrogen loss from paddy fields, a field experiment was conducted by setting single applications of chemical or organic fertilizer, combined organic and inorganic application, and single application of controlled release fertilizer under reduced nitrogen conditions. The results showed that, compared with the conventional fertilization treatment(CF), there was no significant difference in rice grain and straw yield between the single chemical fertilizer treatment(T1) and the organic-inorganic combined treatment(T3); the single organic fertilizer treatment(T2) decreased the rice grain yield by 13.0%, and decreased straw yield by 17.1%; single application of controlled-release fertilizer(T4) increased rice grain and straw yield by 15.7% and 21.0%, respectively. Further, compared with CF, the single application of chemical fertilizer(T1), organic fertilizer(T2), and organic-inorganic combined application(T3) reduced the total nitrogen leaching loss at 30 cm depths by 26.9%, 18.0%, and 33.9%, respectively. The loss of ammonia nitrogen leaching with T1, T2, and T3 decreased by 24.4%, 36.9%, and 36.6%, respectively, and the loss of nitrate nitrogen leaching decreased by 40.2%, 4.8% and 46.4%. The total nitrogen leaching at 60 cm soil depths was reduced by 34.2%, 26.3%, and 42.1%, the loss of ammonia nitrogen leaching was reduced by 31.4%, 35.7%, and 46.6%, and the loss of nitrate nitrogen leaching was reduced by 8.0%, 10.1%, and 23.9% for T1, T2, and T3, respectively. The total nitrogen loss at 30 and 60 cm depths increased by 41.6% and 14.0% in the single application of controlled release fertilizer(T4) treatment. Considering factors such as agronomic and environmental benefits of different fertilization modes, T1 and T3 are suitable environmentally friendly alternative fertilization modes.

Enhanced adsorption of phosphorus in soil by lanthanum-modified biochar: improving phosphorus retention and storage capacity.

Use of soil adsorbent is an effective method for the promotion of phosphorus adsorption capacity of soil, though most of the soil adsorbents have weak phosphorus retention ability. Herein, we compared the traditional gypsum (GP) and zeolite (ZP) adsorbents to explore the phosphorus retention ability of lanthanum modified walnut shell biochar (La-BC) in soil. The results showed that with the increase of exogenous phosphorus concentration, the adsorption amount of phosphorus by adsorbents in soil increased at first and then tended to be stable. The maximum adsorption capacity of soil to phosphorus is gypsum, lanthanum-modified biochar > zeolite, and the addition of lanthanum-modified biochar can improve the adsorption capacity of soil to phosphorus, enhance the binding strength of soil and phosphorus, improve the ability of soil to store phosphorus, reducing phosphorus adsorption saturation, and is beneficial to control the leaching of soil phosphorus. FTIR and XRD analysis showed that the adsorption of phosphorus by each adsorbent in soil was mainly chemical precipitation. The response surface analysis showed that the adsorption performance of La-BC+S was the best when the concentration of exogenous phosphorus was 50.0 mg/L, pH was 6.47, and the reaction time was 436.98 min. This study provides a reference for soil adsorbents to hold phosphorus and reduce the risk of phosphorus leaching to avoid groundwater pollution.

LINC00452 promotes ovarian carcinogenesis through increasing ROCK1 by sponging miR-501-3p and suppressing ubiquitin-mediated degradation.

Ovarian cancer refers to all sorts of cancerous growth that starts from the ovary. Dysregulation of long non-coding RNAs (lncRNAs) is associated with ovarian cancer development and progression. Cellular expression and localization of LINC00452 in ovarian cancer cells were detected by qPCR and FISH. The roles of LINC00452 in ovarian carcinogenesis were characterized by MTT, transwell and colony-formation assays in vitro as well as xenograft mouse model. The underlying mechanism was explored by microarray, RIP, Co-IP and luciferase reporter assays. This study identified a novel lncRNA LINC00452 being elevated in both ovarian cancer cells and tumor tissues in patients. Such aberrant expression of LINC00452 was negatively correlated with relapse-free survival of ovarian cancer patients. Overexpression of LINC00452 potentiated CaOV3 cell viability, migration and invasion in vitro as well as xenograft tumor growth in vivo. Evidence from the current study suggests that the carcinogenicity of LINC00452 is partially due to competitive sponging of miR-501-3p followed with release of repression on the ROCK1, a key effector in Rho signaling pathway. Irrespective of its miRNA sponge function, LINC00452 is capable of preventing ROCK1 protein from ubiquitin/proteasome-mediated degradation via their mutual physical interaction. Our study makes LINC00452 a potential therapeutic target for ovarian cancer.

A Prediction Model for Optimal Primary Debulking Surgery Based on Preoperative Computed Tomography Scans and Clinical Factors in Patients With Advanced Ovarian Cancer: A Multicenter Retrospective Cohort Study.

OBJECTIVE: This study assessed the predictive value of preoperative computed tomography (CT) scans and clinical factors for optimal debulking surgery (ODS) in patients with advanced ovarian cancer (AOC). METHODS: Patients with AOC in International Federation of Gynecology and Obstetrics (FIGO) stage III-IV who underwent primary debulking surgery (PDS) between 2016 and 2019 from nine tertiary Chinese hospitals were included. Large-volume ascites, diffuse peritoneal thickening, omental cake, retroperitoneal lymph node enlargement (RLNE) below and above the inferior mesenteric artery (IMA), and suspected pelvic bowel, abdominal bowel, liver surface, liver parenchyma and portal, spleen, diaphragm and pleural lesions were evaluated on CT. Preoperative factors included age, platelet count, and albumin and CA125 levels. RESULTS: Overall, 296 patients were included, and 250 (84.5%) underwent ODS. The prediction model included age >60 years (P=0.016; prediction index value, PIV=1), a CA125 level >800 U/ml (P=0.033, PIV=1), abdominal bowel metastasis (P=0.034, PIV=1), spleen metastasis (P<0.001, PIV=2), diaphragmatic metastasis (P=0.014, PIV=2), and an RLNE above the IMA (P<0.001, PIV=2). This model had superior discrimination (AUC=0.788>0.750), and the Hosmer-Lemeshow test indicated its stable calibration (P=0.600>0.050). With the aim of maximizing the accuracy of prediction and minimizing the rate of inappropriate explorations, a total PIV ≥5 achieved the highest accuracy of 85.47% and identified patients who underwent suboptimal PDS with a specificity of 100%. CONCLUSIONS: We developed a prediction model based on two preoperative clinical factors and four radiological criteria to predict unsatisfactory debulking surgery in patients with AOC. The accuracy of this prediction model needs to be validated and adjusted in further multicenter prospective studies.

[Evaluation of the Combined Removal of Heavy Metals by Saponin and Citric Acid from Municipal Sewage Sludges and Metal Stability Features].

Sewage sludge contains a high level of nutrients, such as nitrogen, phosphorus, potassium and organic matter, with a high recovery value. The presence of heavy metals in sludges has become a major limiting factor in the utilization of these sludges. Therefore, the removal of heavy metals before land application of sewage sludge has become very necessary. Two municipal sewage sludges (S1 and S2) were collected from municipal wastewater plants in Wuhan city and were studied by oscillation leaching experiments with combination of saponin and citric acid to evaluate their removal efficiencies for Cu, Pb and Zn at different volume ratios (20:1-1:20), ratios of solid to liquid (1:20-1:80), reaction times (0-2880 min), and leaching times (1-4). The heavy metal fractions in the sewage sludges were analyzed before and after leaching to study changes in the stability and mobility of heavy metals by calculating a stability factor[relative bonding strength of heavy metals (IR)] and a mobility factor (MF). The results show that the highest removal efficiency of Cu, Pb, and Zn was 43.16% (S1), 32.45% (S2) and 38.69% (S1), respectively under the removal conditions of a volume ratio (saponin:citric acid) of 5:1, solid-liquid ratio of 1:60, and leaching time of 1440 min. The removal efficiencies of Cu and Pb were significantly enhanced and Zn had a small increase with longer leaching times, which was significantly different after two to three washing times. The highest removal rate of Cu, Pb, and Zn was 78.89% (S1), 77.08% (S2) and 49.39% (S1) after four washing times, respectively. Beyond acid soluble and reducible fractions, other forms of heavy metals have very low removal rates after a single leaching. The removal rates of heavy metal fractions increased by increasing the leaching time, particularly for Pb, which in the residual fraction was significantly increased. The stability and mobility of heavy metals changes after each leaching. For example, the IR of Cu, Pb, and Zn increased to 43.63% (S1), 39.44% (S2) and 32.00% (S1), respectively, and the MF of these heavy metals decreased from 30.19% to 79.45% in the sewage sludges after four washing events.

Possible Epithelial Ovarian Cancer Association with HPV18 or HPV33 Infection.

The present study was conducted to investigate the prevalence of HPV infection in epithelial ovarian cancer (EOC) in Hunan province. DNA samples were collected from paraffin embedded ovarian tissue from 322 patients with EOC, 99 with ovarian benign tumors and 199 normal persons. The polymerase chain reaction and direct sequencing were used to identify the HPV types in the samples. The relationship between the infection of human papillomavirus (HPV) and the epithelial ovarian carcinoma (EOC) was investigated combined with clinical data. The prevalence of HPV18 and HPV33 in EOC group and benign group was higher than in the normal group. HPV18 and HPV33 may play a role in the development of both EOC and ovarian benign tumor and may participate in the development of EOC with traditional risk factors, family history and abortion, possibly exerting synergistic effects..

[Effects of irrigation using dairy effluent on grain yield, phosphorus utilization and distribu- tion in soil profile in winter wheat-summer maize rotation system].

Field experiments of winter wheat-summer maize rotation were conducted in North China Plain irrigation area to explore the effects of wheat season irrigation with dairy effluent on grain yield, phosphorus uptake, accumulative phosphorus usage efficiency and phosphorus accumulation in soil. The results showed that the irrigation with dairy effluent significantly improved the yields of winter wheat and summer maize. With the increasing of P2O5 carried by dairy effluent into soil, winter wheat yield increased at first and then decreased. When the P2O5 increased 137 kg · hm(-2), winter wheat yield increased to the maximum (7646.4 kg · hm(-2)) and the phosphorus utilization rate was the highest (24.8%). But excessive phosphorus decreased the winter wheat yield and phosphorus utilization efficiency. Summer maize yield and phosphorus uptake increased with the increase of P2O5 carried by dairy effluent. The summer maize yield increased by 2222.4-2628.6 kg · hm(-2) and the phosphorus uptake increased by 13.9-21.1 kg · hm(-2) in contrast to the control (CK). Under conventional phosphorus fertilization at 88 kg · hm(-2), and the summer maize yield increased by 2235.0 kg · hm(-2) compared with CK. As the time of irrigation with dairy effluent increasing, the grain yield increased more significantly. The cumulative phosphorus utilization in this rotation system increased year by year. After six seasons of crop harvest, the cumulative phosphorus utilization rate increased into 40.0%-47.7%. Under the experimental condition, two times of irrigation with the dairy effluents in the winter wheat-summer maize rotation system was the best operating mode.

Biological conduits combining bone marrow mesenchymal stem cells and extracellular matrix to treat long-segment sciatic nerve defects.

The transplantation of polylactic glycolic acid conduits combining bone marrow mesenchymal stem cells and extracellular matrix gel for the repair of sciatic nerve injury is effective in some respects, but few data comparing the biomechanical factors related to the sciatic nerve are available. In the present study, rabbit models of 10-mm sciatic nerve defects were prepared. The rabbit models were repaired with autologous nerve, a polylactic glycolic acid conduit + bone marrow mesenchymal stem cells, or a polylactic glycolic acid conduit + bone marrow mesenchymal stem cells + extracellular matrix gel. After 24 weeks, mechanical testing was performed to determine the stress relaxation and creep parameters. Following sciatic nerve injury, the magnitudes of the stress decrease and strain increase at 7,200 seconds were largest in the polylactic glycolic acid conduit + bone marrow mesenchymal stem cells + extracellular matrix gel group, followed by the polylactic glycolic acid conduit + bone marrow mesenchymal stem cells group, and then the autologous nerve group. Hematoxylin-eosin staining demonstrated that compared with the polylactic glycolic acid conduit + bone marrow mesenchymal stem cells group and the autologous nerve group, a more complete sciatic nerve regeneration was found, including good myelination, regularly arranged nerve fibers, and a completely degraded and resorbed conduit, in the polylactic glycolic acid conduit + bone marrow mesenchymal stem cells + extracellular matrix gel group. These results indicate that bridging 10-mm sciatic nerve defects with a polylactic glycolic acid conduit + bone marrow mesenchymal stem cells + extracellular matrix gel construct increases the stress relaxation under a constant strain, reducing anastomotic tension. Large elongations under a constant physiological load can limit the anastomotic opening and shift, which is beneficial for the regeneration and functional reconstruction of sciatic nerve. Better regeneration was found with the polylactic glycolic acid conduit + bone marrow mesenchymal stem cells + extracellular matrix gel grafts than with the polylactic glycolic acid conduit + bone marrow mesenchymal stem cells grafts and the autologous nerve grafts.

[Effects of nitrogen application on N2O flux from fluvo-aquic soil subject to freezing and thawing process].

The experiment was conducted to investigate the effects of different forms of nitrogen application (ammonium, NH4(+) -N; nitrate, NO3(-) -N; and amide-N, NH2-N) and different concentrations (40, 200 and 800 mg/L) on N2O flux from the fluvo-aquic soil in a lab-simulated freezing and thawing process. N2O flux slowly decreased with soil freezing, and then increased slowly with soil thawing, reaching a peak at the initial thawing stage. The average N2O fluxes with addition of NH4(+) -N, NO3(-) -N and NH2 -N are 119.01, 611.61 and 148.22 microg x (m2 x h) (-1) respectively at the concentration of 40 mg/L; 205.28, 1084.40 and 106.13 microg x (m2 x h)(-1) at the concentration of 200 mg/L; 693.95, 1 820.02 and 49.74 microg x (m2 x h)(-1) at the concentration of 800 mg/L. The control is only 100.35 microg x (m2 x h)(-1). N2O flux with addition of NH4(+) -N and NO3(-) -N increased with increasing concentration, ranging from 17.49% to 425.67% for NH4(+) -N, and 563.38% to 1 458.6% for NO3(-) -N compared with control. There was a time lag achieving stable N2O emission flux with concentration increase. In contrast, by adding NH2 -N to soil, N2O flux decreased with increasing concentration. In sum, inorganic nitrogen such as NH4(+) -N or NO3(-) -N fertilizer incorporated in soil enhanced the cumulative N2O flux from the fluvo-aquic soil relative to amide-N. This study suggested that ammonium and nitrate concentration in overwintering water should be less than 200 mg/L and 40 mg/L in order to reduce N2O emissions from soil, regardless of amide-N.

[Inhibitory effects of human papilloma virus 18 E6 gene in HeLa cell transfected with shRNA].

OBJECTIVE: To investigate the inhibition of HPV18E6 gene in HeLa cell transfected with plasmid expressing human papilloma virus 18 E6 (HPV18E6) short hairpin RNA (shRNA). METHODS: We synthesized two HPV18E6 shRNA frames and sub-cloned them into pSUPER which can express shRNA in mammalian cells to construct pE6-1shRNA and pE6-2shRNA which were mutant in E6 shRNA frame. The pE6-1shRNA, pE6-2shRNA and pcDNA3.1 were co-transfected into HeLa cells by cationic liposome respectively and the positive transfectants were selected by G418. The HPV18E6 mRNA and protein expression level was detected by semi-quantitative RT-PCR and streptavidin-peroxidase conjugated method (SP) to assay the inhibitory effects of pE6shRNA. RESULTS: We successfully constructed several new HeLa cell lines transfected with pE6-1shRNA and pE6-2shRNA. In the HeLa cells without transfection and the HeLa cells transfected with pE6-1shRNA plasmid, the HPV18E6 mRNA levels were 1.14 +/- 0.45, 0.76 +/- 0.28 respectively, and the difference of HPV18E6 mRNA levels was significant (P < 0.05). The inhibition efficiency of HPV18E6 gene mRNA was 33.3% and the HPV18E6 protein levels were declined after transfection with pE6-1shRNA. In the HeLa cells transfected with pE6-2shRNA and pSUPER plasmids, HPV18E6 mRNA and protein expression levels were not different from those in wild HeLa cells. CONCLUSIONS: The pE6-1shRNA plasmid can inhibit HPV18E6 expression in HeLa cells, which is persistent, specific and heritable.

Straw bio-degradation by acidogenic bacteria and composite fungi.

A composite microbial system, including a strain of Candida tropicalis (W3), a strain of Lactobacillus plantarm(WY3) and three strains of basidiomycete pL104, pL113 and C33, was chosen to degrade corn straw. The final pH was acid owing to the inoculation of acidogenic bacteria, and under this condition the composite fungi system could produce complex enzyme to destroy the compact structure of corn straw. The experimental results showed that the biomass of composite fungi could reach up to maximum when the pH value was 4.5. Through the bio-degradation by combining acidogenic bacteria with the composite fungi system, the cellulose, hemi-cellulose and lignin degradation rates of corn straw powder were 26.36%, 43.30% and 26.96%, respectively. And the gross crude protein content increased 60.41%. This study provided the evidence for the feasibility of developing a composite microbial system with high capability of degrading straw lignocelluloses in order to make reasonable use of straw resource and protect rural eco-environment.

Electrophysiological effects of anthopleurin-Q on rat hepatocytes.

AIM: To study the effects of AP-Q on CCl(4)-induced acute liver injury, delayed outward potassium current (I(K)), inward rectifier potassium current (I(K1)) and calcium release-activated calcium current (I(CRAC)) in isolated rat hepatocytes. METHODS: A single dose of CCl(4) (10 microg/mL, ip) was injected to induce acute liver injury in rats. Serum aminotransferase activities were determined. Whole cell patch-clamp techniques were used to investigate the effects of AP-Q on delayed outward potassium current (I(K)), inward rectifier potassium current (I(K1)) and calcium release-activated calcium current (I(CRAC)). RESULTS: AP-Q (3.5 and 7 microg/kg) pretreatment significantly reduced ALT and AST activities. AP-Q 0.1-100 nM produced a concentration-dependent increase of I(K) with EC(50) value of 5.55+/-1.8 nM (n=6). AP-Q 30 nM shifted the I-V curve of I(K) leftward and upward. CCl(4) 4 mM decreased I(K) current 28.6+/-6.5% at 140 mV. After exposure to CCl(4) for 5 min, AP-Q 30 nM attenuated the decrease of I(K) induced by CCl(4) close to normal amplitude. AP-Q 0.01-100 nM had no significant effect on either inward or outward components of I(K1) at any membrane potential examined. AP-Q 0.1-100 nM had no significant influence on the peak amplitude of I(CRAC), either, and did not affect the shape of its current voltage curve. CONCLUSION: AP-Q has a protective effect on CCl(4)-induced liver injury, probably through selectively increased I(K) in hepatocytes.

Analysis of Human Platelet Antigen Genotypic Frequencies in Chinese Population by PCR Amplification with Sequence Specific Primers.

In present study, a method for genotyping for human platelet antigen (HPA) systems by means of the polymerase chain reaction amplification with sequence-specific primers (PCR-SSP) technique was developed and employed to determine the human platelet antigen frequencies in the Chinese population. Primers sets were designed to allow PCR amplification for five systems using the same assay conditions. Platelets from 110 random Chinese blood donors were typed for human platelet alloantigens HPA-1 to -5 by the method. The results showed that the HPA genotypic frequencies observed in the 110 donors were 0.91 and 0.09 for HPA-1a and HPA-1b, 0.86 and 0.14 for HPA-2a and HPA-2b, 0.60 and 0.40 for HPA-3a and HPA-3b, 0.92 and 0.08 for HPA-4a and HPA-4b, and 0.85 and 0.15 for HPA-5a and HPA-5b, respectively. In conclusion the method is feasible and practical and may be available to typing for HPA in the clinical laboratories.