Five candidate biomarkers associated with the diagnosis and prognosis of cervical cancer.

PURPOSE: Cervical cancer (CC) is one of the most general gynecological malignancies and is associated with high morbidity and mortality. We aimed to select candidate genes related to the diagnosis and prognosis of CC. METHODS: The mRNA expression profile datasets were downloaded. We also downloaded RNA-sequencing gene expression data and related clinical materials from TCGA, which included 307 CC samples and 3 normal samples. Differentially expressed genes (DEGs) were obtained by R software. GO function analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs were performed in the DAVID dataset. Using machine learning, the optimal diagnostic mRNA biomarkers for CC were identified. We used qRT-PCR and Human Protein Atlas (HPA) database to exhibit the differences in gene and protein levels of candidate genes. RESULTS: A total of 313 DEGs were screened from the microarray expression profile datasets. DNA methyltransferase 1 (DNMT1), Chromatin Assembly Factor 1, subunit B (CHAF1B), Chromatin Assembly Factor 1, subunit A (CHAF1A), MCM2, CDKN2A were identified as optimal diagnostic mRNA biomarkers for CC. Additionally, the GEPIA database showed that the DNMT1, CHAF1B, CHAF1A, MCM2 and CDKN2A were associated with the poor survival of CC patients. HPA database and qRT-PCR confirmed that these genes were highly expressed in CC tissues. CONCLUSION: The present study identified five DEmRNAs, including DNMT1, CHAF1B, CHAF1A, MCM2 and Kinetochore-related protein 1 (KNTC1), as potential diagnostic and prognostic biomarkers of CC.

Interactive effects of multiple climate change factors on ammonia oxidizers and denitrifiers in a temperate steppe.

Global climate change could have profound effects on belowground microbial communities and subsequently affect soil biogeochemical processes. The interactive effects of multiple co-occurring climate change factors on microbially mediated processes are not well understood. A four-factorial field experiment with elevated CO2, watering, nitrogen (N) addition and night warming was conducted in a temperate steppe of northern China. Real-time polymerase chain reaction and terminal-restriction fragment length polymorphism, combined with clone library techniques, were applied to examine the effects of those climate change factors on N-related microbial abundance and community composition. Only the abundance of ammonia-oxidizing bacteria significantly increased by nitrogen addition and decreased by watering. The interactions of watering × warming on the bacterial amoA community and warming × nitrogen addition on the nosZ community were found. Redundancy analysis indicated that the ammonia-oxidizing archaeal community was affected by total N and total carbon, while the community of bacterial amoA and nosZ were significantly affected by soil pH. According to a structural equation modeling analysis, climate change influenced net primary production indirectly by altering microbial abundance and activities. These results indicated that microbial responses to the combination of chronic global change tend to be smaller than expected from single-factor global change manipulations.

[Responses of Soil Ammonia Oxidizers to Simulated Warming and Increased Precipitation in a Temperate Steppe of Inner Mongolia].

Soil ammonia oxidizers, as key players for the ammonia oxidation process in soil N cycling, could respond, adapt, and give feedback to global change. In this research, soil samples were collected from a long-term field experiment with increased precipitation and warming in a temperate steppe of Inner Mongolia. We analyzed the responses of the abundance, diversity, and community structure of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) to warming and increased precipitation using quantitative real-time PCR, terminal restriction fragment length polymorphism (T-RFLP), and clone library. The results showed that increased precipitation significantly stimulated soil pH and warming significantly reduced soil respiration (SR). No significant difference was detected regarding the abundances of amoA genes across all treatments, whereas increased precipitation significantly affected the community structure of soil AOB. However, the interactive effect between warming and increased precipitation had no significant influence on the community structure of soil ammonia oxidizers. The result of the structural equation model indicated that the plant diversity and community structures of soil ammonia oxidizers were significantly correlated, suggesting that there were certain relationships among climate change, microbes, and plants. In conclusion, this study confirmed that soil microorganisms had the ability to adapt to climate change, which could provide important information for predicting future changes in ecosystems.

Morphological Variation and Recovery Mechanism of Residual Crude Oil by Biosurfactant from Indigenous Bacteria: Macro- and Pore-Scale Experimental Investigations.

Microbial enhanced oil recovery (MEOR) is being used more widely, and the biological contributions involved in MEOR need to be identified and quantified for the improvement of field applications. Owing to the excellent interfacial activity and the wide distribution of producing strains in oil reservoirs, lipopeptides have proved to be an essential part of the complex mechanisms in MEOR. In this study, crude lipopeptides were produced by a strain isolated from an indigenous community in an oil reservoir. It was found that crude lipopeptides can effectively reduce the IFT (interfacial tension) to 10(-1)~10(-2) mN/m under high salinity without forming stable emulsions, and the wettability of natural sandstone can be enhanced (Amott index, from 0.36 to 0.48). The results of core flooding experiments indicate that an additional 5.2% of original oil in place can be recovered with a 9.5% reduction of injection pressure. After the shut-in period, the wettability of the core, the reduction of injection pressure, and the oil recovery can be improved to 0.63, 16.2% and 9.6%, respectively. In the microscopic flooding experiments, the crude oil in membrane, cluster, and throat states contribute nearly 90% in total of the additional oil recovery, and the recovery of membranestate oil was significantly enhanced by 93.3% after shut in. Based on the results in macro and pore scale, the IFT reduction and the wettability alteration are considered primary contributors to oil recovery, while the latter was more dominant after one shut-in period.

Abundance and community structure of ammonia-oxidizing Archaea and Bacteria in response to fertilization and mowing in a temperate steppe in Inner Mongolia.

Based on a 6-year field trial in a temperate steppe in Inner Mongolia, we investigated the effects of nitrogen (N) and phosphorus (P) fertilization and mowing on the abundance and community compositions of ammonia-oxidizing Bacteria (AOB) and Archaea (AOA) upon early (May) and peak (August) plant growth using quantitative PCR (qPCR), terminal-restriction fragment length polymorphism (T-RFLP), cloning and sequencing. The results showed that N fertilization changed AOB community composition and increased AOB abundance in both May and August, but significantly decreased AOA abundance in May. By contrast, P fertilization significantly influenced AOB abundance only in August. Mowing significantly decreased AOA abundance and had little effect on AOA community compositions in May, while significantly influencing AOB abundance in both May and August, Moreover, AOA and AOB community structures showed obvious seasonal variations between May and August. Phylogenetic analysis showed that all AOA sequences fell into the Nitrososphaera cluster, and the AOB community was dominated by Nitrosospira Cluster 3. The results suggest that fertilization and mowing play important roles in affecting the abundance and community compositions of AOA and AOB.

Kinetics and computational docking studies on the inhibition of tyrosinase induced by oxymatrine.

A combination of enzymatic inhibition kinetics and computational prediction was employed to search for an effective inhibitor of tyrosinase. We found that oxymatrine significantly inhibited tyrosinase, and that this reaction was not accompanied by detectable conformational changes. Kinetic analysis showed that oxymatrine reversibly inhibited tyrosinase in a mixed-type manner. Measurements of intrinsic and ANS-binding fluorescences showed that oxymatrine did not induce any conspicuous changes in the tertiary structure. We also conducted a docking simulation between tyrosinase and oxymatrine using two docking programs, Dock6.3 and AutoDock4.2 (binding energy was -118.81 kcal/mol for Dock6 and -8.04 kcal/mol for AutoDock4). The results also suggested that oxymatrine interacts mostly with the residues of CYS83 and HIS263 in the active site of tyrosinase. This strategy of predicting tyrosinase inhibition by simulation of docking coupling with kinetics may prove useful in screening for potential tyrosinase inhibitors. Knowledge of tyrosinase inhibition can provide medical, cosmetic, and agricultural applications. Our study suggests that oxymatrine is an important agent for various applications related to pigment formation.

[Gamma event-related synchronization analysis: a new method for cortical function mapping].

OBJECTIVE: To localize the sensory motor cortex of human brain by analyzing the power change in Gamma band (> 60 Hz) of electrocorticography (ECoG) data. METHODS: Eight patients with intractable epilepsy underwent temporary placement of subdural electrodes. After surgery, sensory evoked potential (SEP), electrocortical stimulation (CES) and event-related synchronization analysis of Gamma band (Gamma ERS analysis) were performed to reduce the risk of complications. The results of Gamma ERS analysis were compared with those of SEP and CES. RESULTS: The results of Gamma ERS analysis had 80.7% electrodes fitting perfectly those of CES and SEP. And the percentage reached 92.3% if electrodes were superimposed or added adjacently. CONCLUSION: The Gamma ERS analysis is a new sensitive and precise method for cortical function mapping.

[The responses of moxibustion and the countermeasures].

To make a distinction among the local response of body, the moxibustion sensation, its influence on the disease, adverse reaction and others during and after the moxibustion treatment, and explore the countermeasures to these reactions in order to guide the clinical practice. Of them, the responses of the body surface and local acupoints are usual one of the bases to assess the moxibustion effect, while the occurs of moxibustion sensation and its influence on the disease are normal, which is not necessary to deal with, and the adverse reaction and others could be handled according to the different situations.

Computational prediction of protein-protein interactions of human tyrosinase.

The various studies on tyrosinase have recently gained the attention of researchers due to their potential application values and the biological functions. In this study, we predicted the 3D structure of human tyrosinase and simulated the protein-protein interactions between tyrosinase and three binding partners, four and half LIM domains 2 (FHL2), cytochrome b-245 alpha polypeptide (CYBA), and RNA-binding motif protein 9 (RBM9). Our interaction simulations showed significant binding energy scores of -595.3 kcal/mol for FHL2, -859.1 kcal/mol for CYBA, and -821.3 kcal/mol for RBM9. We also investigated the residues of each protein facing toward the predicted site of interaction with tyrosinase. Our computational predictions will be useful for elucidating the protein-protein interactions of tyrosinase and studying its binding mechanisms.

Capturing amplitude changes of low-frequency fluctuations in functional magnetic resonance imaging signal: a pilot acupuncture study on NeiGuan (PC6).

OBJECTIVES: This study aims to examine amplitude changes of low-frequency oscillations (fALFF) in the blood-oxygen level-dependent (BOLD) signal associated with acupuncture on NeiGuan (PC6). EXPERIMENTAL DESIGN: Ten (10) healthy adults participated in a functional magnetic resonance imaging (i.e., nuclear medicine; fMRI) study. During the brain-imaging procedure, the participants were instructed to lie quietly; they did not perform any cognitive task. MAIN OUTCOME MEASURES: Three (3) fMRI scans were conducted for each participant: a first resting-state scan (R1), a stimulating-acupoint scan (AP), and a second resting-state scan (R2) after AP. Individual fALFF maps were calculated for each scan. RESULTS: During R1, consistent with previous studies, the default network regions showed significantly detectable fALFF amplitudes. Acupuncture on PC6 increased fALFF amplitudes within the anterior cingulate cortex (ACC), occipital fusiform gyrus, posterior cingulate cortex, and precuneus (PCC/PCU). In contrast, during R2, fALFF within PCC is still significantly higher than R1 while ACC and cerebellum showed decreased fALFF. CONCLUSIONS: These findings imply that stimulating PC6 can change the amplitude of the intrinsic cortical activity of the brain. In particular, a continuous and temporally consistent effect of acupuncture within PCC not the common brain circuit of pain including ACC and cerebellum was observed. Considering the cognitive functions and deficits of the relevant areas in mild cognitive impairment and Alzheimer disease, acupuncture on PC6 could potentially affect both psychiatric and neurological disorders. Thus, stimulating PC6 may be a candidate method for improving cognitive impairment.

[Study on catgut implantation therapy compared with Deanxit for treatment of depression].

OBJECTIVE: To compare the therapeutic effect of catgut implantation and Deanxit for treatment of depression. METHODS: Ninety-four cases of depression were randomly divided into a catgut implantation group (50 cases) and a medication group (44 cases). Catgut implantation group was applied with catgut implantation at Hanyan (GB 4), Baihui (GV 20), Shenmen (HT 7), Fenglong (ST 40), Taichong (LR 3), once two weeks, and 4 times in total. Medication group was treated with oral administration of Deanxit, for eight weeks in total. The the-rapeutic effects of both groups were compared with Hamilton Depression-17 Items Scale (HAMD-17) before and after treatment. RESULTS: The scores of HAMD-17 were decreased after treatment in both groups (both P<0.05), and there was no significant difference between two groups after treatment (P>0.05). The total effective rate was 87.8% (43/49) in catgut implantation group and 90.7% (39/43) in medication group, with no significant difference between two groups (P>0.05). CONCLUSION: Catgut implantation can effectively improve the symptoms of depression patients, with a similar therapeutic effect with Deanxit, and it has a better clinical compliance without any adverse effects.

The protective effects of osmolytes on yeast alcohol dehydrogenase conformational stability and aggregation.

The protective effects of four osmolytes (trehalose, dimethysulfoxide, glycine and proline) on the conformational stability and aggregation of guanidine-denatured yeast alcohol dehydrogenase (YADH) have been investigated in this paper. The results show that the four osmolytes protect YADH against unfolding and inactivation by reducing ki (inactivation rate constants), increasing DeltaDeltaGi (transition free energy changes at 25 degrees C), increasing Cm (value for the midpoint of denaturation) and decreasing its ANS-binding fluorescence intensity. Furthermore, these osmolytes can prevent YADH aggregation in a concentration-dependent manner during YADH refolding.

[Correlation analysis on changes between cognitive ability and brain fMRI after acupoint thread embedding in Alzheimer's disease patients].

OBJECTIVE: To explore the mechanism of acupoint thread embedding (ATE) in treating Alzheimer's disease (AD) by means of brain functional magnetic resonance imaging (fMRI). METHODS: Twenty-six patients with mild to moderate AD were assigned to the tested group and the control group in pairing, 13 in each group. ATE was performed in patients of the tested group at Shenmen (HT7), Fenglong (ST40), Taixi (KI3) and Zusanli (ST36) acupoints, once every month for 6 times in total; but for patients of the control group only sham operation was done at the same acupoints. The correlation between cognitive ability and fMRI were assessed by comparing the scores of Mini-Mental State Examination (MMSE) and Alzhemer's Disease Assessment Scale-Cognitive section (ADAS-Cog), monitored at two time points, namely, in one week before starting treatment and 1 week after terminating 6-month treatment. Furthermore, fMRI was detected to obtain the data of the cerebral blood oxygen content before treatment and 6 months after completing the 6-month therapeutic course. Correlation analysis was conducted on the obtained data using software SPM. RESULTS: After treatment, changes of MMSE score significently increased in the test group (P < 0.01) and decreased in the control group (P < 0.01), and ADAS-Cog decreased in the test group (P < 0.01) and increased in the control group (P < 0.05), showing statistical significance between the two groups (P < 0.01, P < 0.05). The activated regions related with the change of MMSE were mainly inferior frontal gyrus, middle frontal gyrus, superior frontal gyrus, transverse temporal gyrus, on the left cerebrum and superior frontal gyrus, middle frontal gyrus, precentral gyrus, hippocampal gyrus, cingulate gyrus, postcentral gyrus, paracentral lobule on the right cerebrum; those related with change of ADAS-Cog were superior frontal gyrus, middle frontal gyrus, fusiform gyrus, inferior temporal gyrus, hippocampal gyrus on the left cerebrum, and superior frontal gyrus, middle frontal gyrus, precentral gyrus, fusiform gyrus, transverse temporal gyrus, postcentral gyrus, middle occipital gyrus on the right cerebrum. CONCLUSION: ATE could improve the cognitive ability of AD patients, its possible mechanism may be through the activation on the cognition related regions of frontal, temporal lobe and marginal system, as well as those of cerebellum.

The inhibition kinetics and thermodynamic changes of tyrosinase via the zinc ion.

We found that Zn(2+) conspicuously inactivated tyrosinase in a mixed-type inhibition manner: the final level of residual activity was abolished at the equilibrium state with concentration of 0.25 mM Zn(2+). Changes of both K(m) and V(max) by various concentrations of Zn(2+) in Lineweaver-Burk plot were observed. To see whether Zn(2+) also induced conformational change of tyrosinase and how thermodynamical changes by ligand binding were occurred, the intrinsic fluorescence studies as well as calorimetric measurements were conducted. The results showed that the Zn(2+) binding to tyrosinase directly induced conformational change of tyrosinase, and the changes of thermodynamic parameters such as enthalpy (DeltaH), Gibbs free-energy (DeltaG), and entropy (DeltaS) were obtained as 60+/-7.0 kJ/mol, -14.54 kJ/mol and 248.53 J/(K mol), respectively. The inactivating effect of Zn(2+) on tyrosinase was completely prevented by incubation with bovine serum albumin, which has a Zn(2+) binding motif in its structure. We suggested that Zn(2+) ligand-binding affected the substrate's accessibility due to the conformational changes and thus, the complex type of inhibition has occurred with the calorimetric changes.

[The epitope analysis of beta Netrin and preparation and characterization of its antibodies].

AIM: To prepare and characterize anti-human beta-Netrin antibodies. METHODS: B cell dominant epitopes of human beta-Netrin C-terminal 114 amino acid sequences were predicated by the GoldKey software. One of the epitopes was synthesized and coupled with bovine serum album (BSA) by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). The BALB/c mice were immunized with the coupled protein. The splenocytes of immunized mice were fused with Sp2/0 cells by routine method and the hybridomas were selected in HAT medium. The hybridoma cells secreting specific antibody were detected by ELISA and cloned by limiting dilution. The titer specificity, and Ig subclass of anti-beta-Netrin mAbs were characterized by ELISA, Western blot and immunocytochemical staining. In addition, New Zealand rabbits were immunized with the coupled protein to prepare polyclonal antibody against beta-Netrin. The specificity of the antiserum was verified by Western blot. RESULTS: A 16-mer peptide NH2-FRGKRTLYPES-WTDRG-COOH was the dominant epitope of the B cells. Synthesized peptide coupled with BSA was used as the immunogen to immunize BALB/c mice. Three hybridoma cell lines that stably secrete specific mAbs were obtained. The result of immunocytochemical staining showed that prepared mAb specifically recognize the antigen in the neuronal cells. The polyclonal antibody against beta-Netrin had high specificity. Western blot analysis showed that the antiserum bind with the prokaryotically expressed beta-Netrin specifically. CONCLUSION: Using the synthesized peptides as hapten, we have prepared epitope-specific mAbs and pAb against beta-Netrin successfully.

[The viral resistance of transgenic tobacco plants containing untranslatable PVYN coat protein gene].

In previous study, we have proved that the resistance of the transgenic tobacco plants containing untranslatable PVYN CP gene was mediated by PVYN CP transgene RNAs, and the resistance mechanism was similar to PTGS. In this paper, T0 progeny transgenic lines with different resistant levels were selected to further study the transgene inheritance and resistant stability of transgenic plants. Results showed that T0 progeny susceptible lines, which contained 1-2 transgene copies, displayed a 3:1 segregation ratio in T1 progeny lines; T0 progeny middle resistant lines, which contained 3-4 transgene copies, revealed a 15:1 segergation ratio in T1 progeny lines; T0 progeny highly resistant lines, which contained 5-7 transgene copies, followed 15:1 or 63:1 segregation pattern in T1 progeny lines. Southern blot analysis revealed the resistance in most T1 and T2 progeny transgenic lines was related to copy numbers of the transgene. Northern blot analysis indicated that PVYN CP transgenes were expressed at transcription level in most T1 and T2 progeny transgenic lines, and the transgene mRNA accumulation in cytoplasm varied among transgenic lines. There was an inverse correlation between transgene transcript accumulation and virus resistance. Resistance of transgenic lines was inheritable over at least two generations, and the resistance of T2 progeny transgenic lines, which containing untranslatable PVYN CP gene, was (1) not overcome by PVYN particle or PVYN RNA; (2) independent of inoculum levels; (3) resistant to either aphid or mechanically transmitted PVYN; (4) not dependent on plant development stages; (5) PVY-specific (i.e., broad-spectrum resistance was not observed).