Inhibition of Cell Apoptosis by Apicomplexan Protozoa-Host Interaction in the Early Stage of Infection.

Apicomplexan protozoa, which are a group of specialized intracellular parasitic protozoa, infect humans and other animals and cause a variety of diseases. The lack of research on the interaction mechanism between Apicomplexan protozoa and their hosts is a key factor restricting the development of new drugs and vaccines. In the early stages of infection, cell apoptosis is inhibited by Apicomplexan protozoa through their interaction with the host cells; thereby, the survival and reproduction of Apicomplexan protozoa in host cells is promoted. In this review, the key virulence proteins and pathways are introduced regarding the inhibition of cell apoptosis by the interaction between the protozoa and their host during the early stage of Apicomplexan protozoa infection. It provides a theoretical basis for the development of drugs or vaccines for protozoal diseases.

Research and Application of High-Pressure Rotary Jet Method in the Seepage Treatment of Heavy Metal Tailing Ponds of Southwest China.

The developed karst caves may become the seepage channels of heavy metal to the soil and underground water in Southwest China. Therefore, it is necessary to apply effective seepage treatments to the base of heavy metal tailing reservoirs. This paper addressed the high-pressure rotary jet technology and slurry systems used in the seepage treatment of the deep tailing sand of the Shenxiandong tailing pond located in Southwest China. In this study, the factors of fluidity, initial and final setting times, compressive strength, and permeability coefficient of the slurry were conducted. The mechanism analysis was investigated by X-ray diffraction (XRD), scanning electron microscope (SEM), and inductively coupled plasma-mass spectrometry (ICP-MS). Three different types of slurry systems were proposed, and the permeability coefficients of the solidification body following 28 days of curing were less than 1 × 10-7 cm/s. The concentrations of Pb and Zn in the slurry system containing bentonite were reduced by 26.2% and 45.7%, respectively. In the presence of slaked lime and fly ash, the concentrations of Pb and Zn could be reduced by 26.8% and 30%, respectively. A total of 2142 high-pressure rotary jet piles were completed by the high-pressure rotary jet method in the field trial. The diffusion radius of these piles was over 1 m. Following 28 days of curing, the solidification body's compressive strength was 7.45 MPa and the permeability coefficient was 6.27 × 10-8 cm/s. Both the laboratory and on-site trials showed that this method produced a good pollution barrier effect, which could prevent the diffusion of heavy metal into the adjacent underground water through the karst caves. It is also an effective way of engineering technology concerning heavy metal pollution control that occurs in tailing ponds.

Effect of snow cover on water and heat transfer in alpine meadows in the source region of Yellow River.

Extreme snowfall events have been increasing in the Tibetan Plateau, causing greater variations in the snow cover conditions. However, the soil water-heat transfer under different snow conditions has rarely been characterized in detail. Here, by using the multi-source observation data of five years, we analyzed the influences of snow cover on water-heat transfer in alpine meadows of the source region of the Yellow River. The main findings are as follows: In the deep soil, the yearly warming rate from spring to summer was much faster than the cooling rate from autumn to winter, while in the shallow soil, conversely, the former was slower than the latter. Snow cover not only decreased the average soil temperature but also inhibited the occurrence of extremely low temperatures in the soil. The insulation effect of snow was mainly in the mid-frozen period. It was insufficient to balance out the heat lost by the high albedo during early and late frozen periods. In years with more snow, different depths of the soil featured similar thawing dates and plenty of soil voids due to small solid water content and high gravel content, together creating favorable conditions for the snowmelt infiltration, which passed through the frozen layer and infiltrated into the soil of 3.20 m or deeper. In years with less snow, the long-term freezing-thawing cycles aggravated the evaporation and loss of surface soil water in spring. Under different snow cover conditions, the difference in the sensible heat flux was much larger than the latent heat flux in winter and early spring. This study provides a refined physical image of soil water-heat transfer under extreme snow cover conditions in the Tibetan Plateau, which is expected to light the snow cover-frozen soil interaction in the mid-latitude and high-elevation areas.

The role of demographic compensation in stabilising marginal tree populations in North America.

Demographic compensation-the opposing responses of vital rates along environmental gradients-potentially delays anticipated species' range contraction under climate change, but no consensus exists on its actual contribution. We calculated population growth rate (λ) and demographic compensation across the distributional ranges of 81 North American tree species and examined their responses to simulated warming and tree competition. We found that 43% of species showed stable population size at both northern and southern edges. Demographic compensation was detected in 25 species, yet 15 of them still showed a potential retraction from southern edges, indicating that compensation alone cannot maintain range stability. Simulated climatic warming caused larger decreases in λ for most species and weakened the effectiveness of demographic compensation in stabilising ranges. These findings suggest that climate stress may surpass the limited capacity of demographic compensation and pose a threat to the viability of North American tree populations.

Drivers of 2013-2020 ozone trends in the Sichuan Basin, China: Impacts of meteorology and precursor emission changes.

The Sichuan Basin (SCB) of China is known for excessive ozone (O3) pollution owing to high anthropogenic emissions combined with terrain-induced poor ventilation and weak wind fields against the surrounding mountains. While O3 pollution has emerged as a prominent concern in southwestern China yet variations in O3 levels during 2013-2020 are still unclear and the dominant factor in explaining the long-term O3 trend throughout the SCB remains elusive due to uncertainties in emission inventory and variability associated with meteorological conditions. Here, we use extensive basin-wide ambient measurements to examine the spatial pattern and trend of O3 and leverage OMI and TROPOMI satellites in conjunction with MEIC emission inventory to track emission changes. Sensitivity simulations are conducted by using WRF-CMAQ model to investigate the impacts of meteorological variability and emission changes on O3 changes over 2013-2020. O3 concentrations exhibit obvious interannual increases during 2013-2019 and a slight decrease in 2020. Both decreases in the MEIC emission inventory (-2.9% yr-1) and OMI NO2 column density (-3.1% yr-1) reflects the declining trend in NOx emissions over 2013-2020, while anthropogenic VOCs were not adequately regulated during 2013-2017, which explained the majority of deteriorated O3 pollution from 2013 to 2017. Furthermore, attribution analysis based on CMAQ simulations indicate that the unexpected aggravated O3 levels in 2019 is not only modulated by disproportional reductions in VOCs and NOx emissions, but also associated with unfavorable meteorological conditions featured by profound heatwaves and frequent stagnant conditions. In 2020, the abnormal meteorological conditions in May leads to substantial increase of O3 by 26.8 µg m-3 as compared to May 2019, while the considerable enhancement was fully offset by low O3 levels over the whole period which attributes to substantial emission reductions. This study reveals the long-term trend of O3 levels and precursor emissions and highlights the effects of meteorological variability and emission changes on O3 pollution over the SCB, with strong implications for designing effective O3 control measures.

Metal-free C(sp3)-H functionalization of sulfonamides via strain-release rearrangement.

With the increasing awareness of sustainable chemistry principles, the development of an efficient and mild strategy for C(sp3)-H bond activation of nitrogen-containing compounds without the utilization of any oxidant and metal is still highly desired and challenging. Herein, we present a metal-free reaction system that enables C-H bond functionalization of aliphatic sulfonamides using DABCO as a promoter under mild conditions, affording a series of α,ß-unsaturated imines in good yields with high selectivities. This protocol tolerates a broad range of functionalities and can serve as a powerful synthetic tool for the late-stage modification of complex compounds. More importantly, control experiments and detailed DFT calculations suggest that this process involves [2 + 2] cyclization/ring-cleavage reorganization, which opens up a new platform for the establishment of other related reorganization reactions.

Direct Synthesis of α-Amino Nitriles from Sulfonamides via Base-Mediated C-H Cyanation.

Herein, we disclose a transition-metal-free reaction system that enables α-cyanation of sulfonamides through C-H bond cleavage for the preparation of α-amino nitriles, including difficult-to-access all-alkyl α-tertiary scaffolds. More than 50 substrate examples prove a wide functional group tolerance. Additionally, its synthetic practicality is highlighted by gram-scalability and the late-stage modification of natural compounds. Mechanistic experiments suggest that this process involves in situ formation of an imine intermediate via base-promoted elimination of HF.

Origin of regional springtime ozone episodes in the Sichuan Basin, China: Role of synoptic forcing and regional transport.

The Sichuan Basin (SCB) located in southwestern China has long been considered the most polluted city cluster with exposure to unhealthy levels of ozone (O3) at times. However, the features of O3 regional transport and source contributions in SCB are poorly understood. In this study, ambient measurements, ERA5 reanalysis dataset, IASI O3 column, and the Weather Research and Forecasting-Community Multiscale Air Quality (WRF-CMAQ) modeling system coupled with the Integrated Source Apportionment Method (ISAM) module were used to investigate the formation mechanism and sources of a severe O3 episode in spring 2020 over the SCB. In the first stage of the O3 episode, a high-pressure system persisted over the western SCB and caused northeasterly wind fields, leading to enhanced regional transport from the northern boundary with the O3 contribution from the boundary exceeding 50% across the SCB. As the synoptic pattern evolved, southeasterly winds dominated the SCB and the stagnant zone over the Chengdu Plain confined O3 originating from the southern SCB and Chongqing city, leading to the accumulation of precursors and elevated O3 levels. During the O3 episode, transportation and industrial sources were major contributors to O3 formation especially for the Chengdu Plain and Chongqing city. In addition, the O3-rich air mass in the nocturnal residual layer that formed over Chongqing city was transported to the Chengdu Plain through southeastern corridor at 400-1600m above ground-level under the prevailing southeasterly winds. With sunrise and the development of the atmospheric boundary layer, the O3-rich air mass in the residual layer (RL) was entrained to the ground-level via vertical mixing, which further enhanced O3 pollution across the Chengdu Plain. Our results revealed the mechanism of regional transport via northeastern and southeastern corridors during an O3 episode and demonstrated the need for joint emission regulation across the SCB to mitigate O3 pollution.

Patterns and environmental drivers of greenhouse gas fluxes in the coastal wetlands of China: A systematic review and synthesis.

Coastal wetlands play an increasingly important role in regulating greenhouse gas (GHG) fluxes and thus affecting climate change. However, the overall magnitude, trend, and environmental drivers of GHG fluxes in these wetlands of China remain uncertain. Herein, we synthesized data from 70 publications involving 187 field observations to identify patterns and drivers of GHG fluxes across coastal wetlands in China. Average methane (CH4), nitrous oxide (N2O) fluxes, and carbon dioxide (CO2) emissions (ecosystem respiration) across coastal wetlands were estimated as 2.20±0.31 mg·m-2·h-1, 16.44±2.96 µg·m-2·h-1, and 388.76±42.28 mg·m-2·h-1, respectively. GHG emissions varied with tidal inundation, where CH4 and CO2 emissions during tidal inundation were lower than during ebbing. CH4 and CO2 emissions from wetlands decreased linearly with increasing latitude, while N2O did not. CH4 fluxes were positively related to air temperature and aboveground biomass, and CO2 emissions were positively related to soil organic carbon. N2O fluxes were lower with increasing soil pH, and CH4 and CO2 emissions were greater with increasing soil moisture. Based on the results of sustained-flux global warming potential and sustained-flux global cooling potential models, our paper indicate that the fluxes of CH4 and N2O in coastal wetlands have a positive feedback to global warming, which is mainly driven by the CH4 emission. Our synthesis improved understanding of the roles of coastal wetlands in the ecosystem C cycle under global change. We suggest that long-term field observations of GHG fluxes across a wider range of spatiotemporal scales are urgently required to improve the prediction accuracy in GHG fluxes and the assessment of net GHG balance and its contribution to the GWP of coastal wetlands.

Time-course analysis of nuclear events during conjugation in the marine ciliate Euplotes vannus and comparison with other ciliates (Protozoa, Ciliophora).

Ciliates represent a morphologically and genetically distinct group of single-celled eukaryotes that segregate germline and somatic functions into two types of nuclei and exhibit complex cytogenetic events during the sexual process of conjugation, which is under the control of the so-called "mating type systems". Studying conjugation in ciliates may provide insight into our understanding of the origins and evolution of sex and fertilization. In the present work, we studied in detail the sexual process of conjugation using the model species Euplotes vannus, and compared these nuclear events with those occurring in other ciliates. Our results indicate that in E. vannus: 1) conjugation requires about 75 hours to complete: the longest step is the development of the new macronucleus (ca. 64h), followed by the nuclear division of meiosis I (5h); the mitotic divisions usually take only 2h; 2) there are three prezygotic divisions (mitosis and meiosis I and II), and two of the eight resulting nuclei become pronuclei; 3) after the exchange and fusion of the pronuclei, two postzygotic divisions occur; two of the four products differentiate into the new micronucleus and macronucleus, respectively, and the parental macronucleus degenerates completely; 4) comparison of the nuclear events during conjugation in different ciliates reveals that there are generally three prezygotic divisions while the number of postzygotic divisions is highly variable. These results can serve as reference to investigate the mating type system operating in this species and to analyze genes involved in the different steps of the sexual process.

Pr-SNTX, a short-chain three-finger toxin from Papuan pigmy mulga snake, is an antagonist of muscle-type nicotinic acetylcholine receptor (α2ßδε).

Three-finger toxins (3FTxs) are one of the major components in snake venoms. In this study, we isolated a cDNA encoding a short-chain 3FTx, Pr-SNTX, from Pseudechis rossignolii. The amino acid sequence of Pr-SNTX is nearly identical to that of its ortholog in Pseudechis australis. Pr-SNTX protein inhibited muscle-type (α2ßδε), but not neuronal α7 nicotinic acetylcholine receptor (nAChR) activity.

[Comparison of early clinical effects between Activ C cervical disc replacement and anterior cervical discectomy and fusion for single-level cervical spondylosis].

OBJECTIVE: To compare the early clinical effects of Activ C cervical disc replacement (ACDR) and anterior cervical discectomy and fusion (ACDF) in treating single-level cervical spondylosis. METHODS: The clinical data of 76 patients with single-level cervical spondylosis underwent surgery from July 2009 to September 2012 were retrospectively analyzed. Among them, 28 patients were treated with ACDR (ACDR group), including 18 males and 10 females, aged from 32 to 62 years old with an average of (45.2±6.2) years; and 48 patients were treated with ACDF (ACDF group), including 28 males and 20 females, aged from 33 to 60 years old with an average of (45.8±6.4) years. Visual analogue scale (VAS), Japanese Orthopedics Association (JOA) score, Short Form-36 (SF-36), imaging data were used to assess the clinical effects after operation. RESULTS: A total of 76 patients were followed up from 6 to 24 months with an average of 13.2 months. VAS of neck pain and brachialgia were improved in all patients after operation (P<0.05), there was no significant difference between two group (P>0.05). Somato-score and psycho-score of SF-36 of two groups were obviously increased (P<0.05), ACDR group was better than that of ACDF group (P<0.05). In ACDR group, there was no significant difference in the range of motion of surgical segments and adjacent segments between preoperative and postoperative (P>0.05); heterotopic ossification around the edge of vertebral body occurred in 1 case on the 6th month after operation, no fusion was found on the 1st year after operation. In ACDF group, the adjacent vertebral disease occurred in 1 case and the patient underwent the reoperation. CONCLUSION: Activ C cervical disc replacement can reduce the degeneration of adjacent segments and its early outcomes for the treatment of single-level cervical spondylosis are satisfactory, but the long-term effects still need study.

Cloning the sterol carrier protein 2 genes of Japanese toad (Bufo japonicus formosus) and Chinese toad (Bufo gargarizans) and its tissue expression analysis.

In this study, to clarify the bioactive polypeptides included in the skins and secretions of Bufo, we screened the Japanese toad (Bufo japonicus formosus) skin cDNA liary by colony polymerase chain reaction (PCR), and obtained a transcript of 1 075 bp consisting of 1 37 bp 5' untranslated region (UTR), 515 bp 3' UTR and a 423 bp open reading frame (ORF) encoding a polypeptide of 140 amino acid residues (GenBank accession number: KF359945). Homolog analysis showed a 70%-96% homology with sterol carrier protein-2 (SCP-2) present in other animals, which is implicated in lipid metabolism of other organisms. The gene SCP-2 of Chinese toad (B. gargarizans) was cloned from a first strand cDNA of Bufo skin (GenBank accession number: KF381341) via PCR, whose encoding polypeptide has only one amino acid difference from that of Japanese toad. Tissue distribution analysis showed that SCP-2 expressed in all organs tested, though in the liver and spleen it manifested lower expression than in other organs. These findings might indicate SCP-2 being one of the active ingredients in toad skin. These findings may in turn have implications for further drug development from traditional Chinese medicine sources.

[Cloning and bioinformatic analysis of TAGLN2 cDNA of Bufo japonicus formosus].

To study the bioactive polypeptides included in Bufo skin and its secretions the plasmid skin cDNA library of adult Japanese toad Bufo japonicus formosus was prepared. The pSD64TR has been used as the vector and the cloning sites are Xho I and EcoR I. To screen cDNAs encoding bioactive components, the plasmid cDNA library was transformed into E. coli DH5 competent cells, and positive colonies were screened by colony PCR (polymerase chain reaction). The suspension of a single colony in LB medium was used as the template, SP6 (the upstream primer of the plasmid cDNA library) and a primer with Xho I site and polyT were used as the primers. As the result, 465 positive colonies out of 1 344 were obtained and their plasmid were collected and sequenced. By homologous analysis, it was found that one of the cDNAs encoding a peptide with high homolog with transgelin-2, which was registered in GenBank (accession number: JX197456), and it was indicated as a partial cDNA sequence with a deletion at the 5' end. The transcript is 997 bp consisting of 31 bp 5', 618 bp 3' untranslated region (UTR) and an open reading frame (ORF) of 348 bp encoding a polypeptide of 115 amino acids. In the putative protein product, there is a calponin homology domain, two cysteine residues for a disulfide bond and three a-helix domains, and five potential phosphorylation sites. The homologous analysis indicates 90% similarity with Xenopus (Silurana) tropicalis and 89% with Xenopus laevis, and 71%-85% with other species.

[Bufo gargarizans mcl-1 cloning and its prokaryotic recombinant protein expression].

MCL-1 is encoded by myeloid cell leukemia-1 gene (mcl-1), which is one of the anti-apoptotic members of bcl-2 cell apoptotic gene superfamily. ChanSu is made of dorsal secretions of several Bufo species, commonly used in the prescriptions of traditional Chinese medicine for treating many diseases including cancer. To clarify if mcl-1 is expressed in the dorsal skin of B. gargarizans, the PCR (polymerase chain reaction) was performed with its dorsal skin first strand cDNA as the template and a pair of specific primers of mcl-1, and PCR products were cloned into the pGM-T vector. DNA sequencing indicated that the ORF length was 639 bp encoding 212 amino acid residues, and the homology of 44%-95% with the MCL-1 of several other animals. For the further studies on MCL-1 biological functions during the oncogenesis and preparation of its antibody, the prokaryotic expression construct of pET-28b-mcl-1 was prepared which was confirmed by DNA sequencing, and its recombinant protein expression (0.02% wet weight) in E. coli BL21 (DE3) strain was confirmed by SDS-PAGE and Western blotting.

Stage-specific appearance of cytoplasmic microtubules around the surviving nuclei during the third prezygotic division of Paramecium.

There are six micronuclear divisions during conjugation of Paramecium caudatum: three prezygotic and three postzygotic divisions. Four haploid nuclei are formed during the first two meiotic prezygotic divisions. Usually only one meiotic product is located in the paroral cone (PC) region at the completion of meiosis, which survives and divides mitotically to complete the third prezygotic division to yield a stationary and a migratory pronucleus. The remaining three located outside of the PC degenerate. The migratory pronuclei are then exchanged between two conjugants and fuse with the stationary pronuclei to form synkarya, which undergo three successive divisions (postzygotic divisions). However, little is known about the surviving mechanism of the PC nuclei. In the current study, stage-specific appearance of cytoplasmic microtubules (cMTs) was indicated during the third prezygotic division by immunofluorescence labeling with anti-alpha tubulin antibodies surrounding the surviving nuclei, including the PC nuclei and the two types of prospective pronuclei. This suggested that cMTs were involved in the formation of a physical barrier, whose function may relate to sequestering and protecting the surviving nuclei from the major cytoplasm, where degeneration of extra-meiotic products occurs, another important nuclear event during the third prezygotic division.

New details indicated by different stainings during conjugation of ciliated protozoa Paramecium.

During conjugation of Paramecium caudatum, nuclear events occur in a scheduled program. Morphological studies on nuclear behavior during conjugation of P. caudatum have been performed since the end of the 19th century. Here we report on new details concerning the conjugation of P. caudatum through the staining of conjugating cells with protargol, carbol fuchsin solution, Hoechst 33342 and immunofluorescence labeling with monoclonal antibody of anti-α tubulin. 1) The crescent nucleus is a characteristic of the meiotic prophase of P. caudatum, has an unstained area. We stained this area with protargol, which was separated from the chromatin area and was not detected by the other stainings. 2) In regards to the four meiotic products, it has long been considered that only one product enters the paroral cone region (PC) and survives after meiosis. However, our protargol and immunofluorescence labeling results indicated that PC entrance of the meiotic product happened before the completion of meiosis instead of after. 3) In our previous study, protargol staining indicated the presence of a swollen structure around the central part of the "U" and "V" shaped spindles connecting the two types of prospective pronuclei. However, immunofluorescence labeling with anti-α tubulin antibodies gave a different image from protargol. All these observations form the basis for further studies of their molecular mechanisms.

Localization of stationary pronuclei during conjugation of Paramecium as indicated by immunofluorescence staining.

After the third prezygotic division during conjugation of Paramecium caudatum, migratory and stationary pronuclei are produced. The migratory pronuclei remain in the paroral region tightly against the conjugating boundaries; while the stationary pronuclei are located beside the migratory pronuclei. To date, however, it is not clear what causes this close side-by-side localization between migratory and stationary pronuclei. In the current study, immunofluorescence staining with monoclonal antibody of anti-α tubulin indicated that ''U'' or ''V'' shaped spindles connected the migratory and stationary pronuclei during the third prezygotic division. This observation accounts for the close localization between these two types of pronuclei.

Dynamic behaviour of stationary pronuclei during their positioning in Paramecium caudatum.

During conjugation of Paramecium caudatum, there are two well-known stages when nuclear migration occurs. What happens to the nuclei is closely related to their localisations in cells. The first of these stages is the entrance of one meiotic product into the paroral region. This nucleus survives, while the remaining three outside this area degenerate. The second stage is the antero-posterior localisation of eight synkaryon division products. Four posterior nuclei are differentiated into macronuclear anlagen, whereas four anterior nuclei remain as the presumptive micronuclei. In this experiment, the process of the third prezygotic division of P. caudatum was studied with the help of protargol staining. Here, a third nuclear migration was discovered. By two spindle turnings and two spindle elongations, stationary pronuclei were positioned near migratory pronuclei. This positioning of stationary pronuclei could shorten the distance for transferred migratory pronuclei to recognise and reach the stationary pronuclei. This fosters the synkaryon formation of P. caudatum.

Morphological apoptotic characteristics of the post-meiotic micronuclei in Paramecium caudatum.

In a previous study, the apoptotic degeneration of meiotic products outside the paroral region of Paramecium caudatum was indirectly demonstrated by means of "apofluor" staining. In this experiment, conjugating pairs and exconjugants of P. caudatum were stained with either "apofluor" or carbol fuchsin or both to find some direct evidence to demonstrate the apoptotic characteristics of this process. As a result, asynchronous meiotic nuclear degeneration was observed. Furthermore, a number of additional meiotic nuclei were found. Disintegrating/dividing meiotic nuclei outside the paroral region were observed, which might be the origin of these additional meiotic nuclei. Condensed chromatin and disintegrated chromatin attached to the nuclear membrane were also observed in degenerating nuclei, which are the typical morphological characteristics of apoptosis. Comparison of the cells stained by the above two methods indicated that "apofluor"-stained meiotic nuclei could not be detected by carbol fuchsin in some cells, which suggests a time lag between meiotic nuclear DNA degradation and their eventual disappearance. In this study, some direct evidence was found to show that the meiotic nuclear degeneration in P. caudatum is of apoptotic nature, which further confirmed our previous study (Yang et al. 2007) and indicated that morphological apoptotic characteristics discovered in multicellular organisms do exist in unicellular eukaryotic ciliate protozoa.