Identification of Key Genes Associated with 1,2,6-Tri-O-galloyl-ß-D-glucopyranose Accumulation in Camellia sinensis Based on Transcriptome Sequencing.

Hydrolyzed tannin 1,2,6-tri-O-galloyl-ß-D-glucopyranose (1,2,6-TGGP) possesses significant medicinal properties. However, little is known about its underlying molecular mechanisms. In this study, the levels of 1,2,6-TGGP in tea materials from different cultivars and leaf positions were compared. Additionally, one leaf and one bud sample from six tea cultivars with significant variations in 1,2,6-TGGP levels were analyzed using transcriptome high-throughput sequencing to identify the genes that are responsible for 1,2,6-TGGP accumulation. The sequencing results were mapped to the reference tea genome, revealing a total of 2735 differentially expressed genes (DEGs). This set included four UDP glycosyltransferase (UGTs) and six serine carboxypeptidases-like (SCPLs) genes. Among them, the upregulated SCPLs (CSS0032817) may directly participate in the acylation reaction of 1,2,6-TGGP. In addition, several classes of DEGs, including cytochrome P450, were significantly associated with the 1,2,6-TGGP content, which is potentially involved in their regulation. Overall, these results provide new insights into the molecular mechanism of 1,2,6-TGGP accumulation.

Seroprevalence and Risk Factors of Chlamydia Infection in Cattle in Shanxi Province, North China.

The information on Chlamydia infection in cattle is limited in Shanxi Province, north China. This study aimed to investigate the seroprevalence and risk factors of Chlamydia and Chlamydia abortus infection in cattle in Shanxi Province. In November 2020, a large-scale investigation of Chlamydia seroprevalence was conducted on 981 cattle serum samples collected from 40 cattle farms in 11 cities of Shanxi Province. The seroprevalence of Chlamydia and C. abortus was examined by indirect hemagglutination assay (IHA) and enzyme-linked immunosorbent assay (ELISA), respectively. The seroprevalence of Chlamydia and C. abortus was 52.29% (513/981) and 2.96% (29/981), respectively, in cattle in Shanxi Province. Location was identified as a risk factor for Chlamydia and C. abortus infection (p < 0.05). Under different management patterns, the seroprevalence of Chlamydia and C. abortus in large-scale animal farming companies was higher than that in household animal farms and animal farming cooperatives, and only the seroprevalence of Chlamydia was significantly different in different management patterns (p < 0.01). The results showed that there was higher seroprevalence of Chlamydia in cattle in Shanxi Province, while C. abortus was not the dominant species. This study provided baseline information on Chlamydia infection in cattle in Shanxi Province, which constitutes valuable data for monitoring livestock health and preventing potential zoonoses.

Seroprevalence of Neospora caninum infection and associated risk factors in cattle in Shanxi Province, north China.

Neospora caninum is an obligate intracellular parasitic protozoan that can cause abortions in cattle and pose considerable economic losses to the cattle industry. As a major livestock province, little is known of N. caninum infection in cattle in Shanxi Province, north China. In order to investigate the seroprevalence of N. caninum in cattle in Shanxi Province, 978 cattle serum samples were collected from 11 cities in three representative geographical locations in Shanxi Province, and the N. caninum-specific IgG antibodies were examined using an indirect enzyme linked immunosorbent assay (ELISA) kit commercially available. The results showed that 133 of the 978 examined cattle serum samples (13.60%, 95% CI = 11.45-15.75) were positive for N. caninum antibodies, and the seroprevalence in different cities ranged from 0 to 78.89%. The geographical location and management mode were the risk factors associated with N. caninum infection in cattle herds in Shanxi Province. Cattle in Northern and Central Shanxi Province as well as cattle whose management mode is that of large-scale cattle farming companies are more susceptible to N. caninum infection. This was the first large-scale survey of N. caninum seroprevalence and assessment of associated risk factors in cattle in Shanxi Province, which provided baseline information for the prevention and control of N. caninum infection in cattle in Shanxi Province, north China.

The Seroprevalence of Chlamydia Infection in Sheep in Shanxi Province, China.

Chlamydia, an obligate intracellular bacterium, can cause chlamydiosis in humans and animals worldwide and also leads to serious economic losses to the sheep industry. However, the information on Chlamydia infection in sheep was limited in Shanxi Province, northern China. In the present study, a total of 984 serum samples of sheep were collected from 11 regions in Shanxi Province, northern China in the autumn of 2020. The antibodies against Chlamydia and Chlamydia abortus were examined by the indirect hemagglutination assay (IHA) and indirect enzyme-linked immunosorbent assay (ELISA), respectively. The result showed that 351 (35.67%, 95% CI 32.68-38.66) of 984 serum samples were positive for Chlamydia, and the seroprevalence ranged from 6.67% to 70.79% among the different regions. In addition, antibodies to C. abortus infection were detected in 78 (7.93%, 95% CI 6.24-9.61) of 984 serum samples, and the seroprevalence ranged from 6.24% to 14.81% among the different regions. This is the first report on the seroprevalence of Chlamydia and C. abortus in sheep in Shanxi province, northern China. The findings provide baseline information for preventing and controlling Chlamydia infection in sheep in Shanxi Province, China.

Functional identification of purine permeases reveals their roles in caffeine transport in tea plants (Camellia sinensis).

Caffeine is a characteristic secondary metabolite in tea plants. It confers tea beverage with unique flavor and excitation effect on human body. The pathway of caffeine biosynthesis has been generally established, but the mechanism of caffeine transport remains unclear. Here, eight members of purine permeases (PUPs) were identified in tea plants. They had diverse expression patterns in different tissues, suggesting their broad roles in caffeine metabolism. In this study, F1 strains of "Longjing43" ♂ × "Baihaozao" ♀ and different tea cultivars were used as materials to explore the correlation between caffeine content and gene expression. The heterologous expression systems of yeast and Arabidopsis were applied to explore the function of CsPUPs. Correlation analysis showed that the expressions of CsPUP1, CsPUP3.1, and CsPUP10.1 were significantly negatively correlated with caffeine content in tea leaves of eight strains and six cultivars. Furthermore, subcellular localization revealed that the three CsPUPs were not only located in plasma membrane but also widely distributed as circular organelles in cells. Functional complementation assays in yeast showed that the three CsPUPs could partly or completely rescue the defective function of fcy2 mutant in caffeine transport. Among them, transgenic yeast of CsPUP10.1 exhibited the strongest transport capacity for caffeine. Consistent phenotypes and functions were further identified in the CsPUP10.1-over-expression Arabidopsis lines. Taken together, it suggested that CsPUPs were involved in caffeine transport in tea plants. Potential roles of CsPUPs in the intracellular transport of caffeine among different subcellular organelles were proposed. This study provides a theoretical basis for further research on the PUP genes and new insights for caffeine metabolism in tea plants.

Chinese herbal medicine injections (CHMIs) for chronic pulmonary heart disease: A protocol for a Bayesian network meta-analysis.

BACKGROUND: Chinese herbal medicine injections (CHMIs) are frequently used for various refractory diseases including chronic pulmonary heart disease (CPHD). However, due to the diversity of CHMIs treatments, its relative effectiveness and safety remain unclear. In our study, Bayesian network meta-analysis will be used to identify differences in efficacy and safety between diverse CHMI for CPHD. METHODS: Relevant randomized controlled trials (RCTs) and prospective controlled clinical trials published in PubMed, Google Scholar, Excerpt Medica Database, Medline, Cochrane Library, Web of Science, China Scientific Journal Database, China National Knowledge Infrastructure, Chinese Biomedical Literature Database and Wanfang Database will be systematic searched to identify eligible studies from their establishment to December 2020. The methodological qualities, including the risk of bias, will be evaluated using the Cochrane risk of bias assessment tool. Stata14.2 and WinBUGS 1.4.3 software were used for data synthesis. The evidentiary grade of the results will be also evaluated using the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach. RESULTS: The results of this study will be published in a peer-reviewed journal, and provide reliable evidence for different CHMIs on CPHD. CONCLUSIONS: The findings will provide reference for evaluating the efficacy and safety of different CHMIs for CPHD, and provide a helpful evidence for clinicians to formulate the best adjuvant treatment strategy for CPHD patients. TRIAL REGISTRATION NUMBER: INPLASY2020120004.

Inhibition of EZH2 attenuates coronary heart disease by interacting with microRNA-22 to regulate the TXNIP/nuclear factor-κB pathway.

NEW FINDINGS: What is the central question of this study? The relevance of microRNA-22 (miR-22) has been indicated in coronary heart disease (CHD). How does it exert a protective role in CHD? What is the main finding and its importance? EZH2 inhibited transcription of the miR-22 promoter, thus modulating cell proliferation in human umbilical vein endothelial cells and vascular smooth muscle cells to induce CHD. ABSTRACT: MicroRNA-22 (miR-22) was indicated to modulate cell proliferation in human umbilical vein endothelial cells (HUVECs) under exposure to environmental toxicants. In the present study, we investigated the involvement of miR-22 in the mediation of HUVEC and vascular smooth muscle cell (VSMC) function, hence in the development of coronary heart disease (CHD). miR-22 expression was reduced in serum of CHD patients. Restoration of miR-22 decreased the proliferation, migration and invasion of VSMCs and increased apoptotic cells and inflammatory factors. In contrast, upregulation of miR-22 led to opposite trends in HUVECs. Chromatin immunoprecipitation and dual-luciferase assays validated that enhancer of zeste 2 polycomb repressive complex 2 subunit (EZH2) inhibited transcription of miR-22 promoter. EZH2, overexpressed in serum from CHD patients, diminished VSMC apoptosis, but facilitated HUVEC apoptosis. Luciferase reporter assays confirmed that thioredoxin-interacting protein (TXNIP) was a new direct target of miR-22. Overexpression of TXNIP blocked the function of miR-22 in HUVECs and VSMCs. Taken together, these findings will shed light on the role and mechanism of EZH2 in viability, migration, invasion and apoptosis via the miR-22/TXNIP axis in VSMCs and HUVECs, which might provide new insights into the treatment of CHD.

Nitrate accumulation and leaching potential is controlled by land-use and extreme precipitation in a headwater catchment in the North China Plain.

Nitrate in groundwater is increasing in hilly areas of the world due to diverse land-uses and intensified anthropogenic activity. However, the key factors that control nitrate in groundwater as it leaches through the thin vadose zone are still poorly understood. In this study, the behavior of nitrate in the vadose zone during a normal year (2015) and a wet year (2016) were investigated in a cultivated farmland (FL) under wheat-maize double cropping and a field of natural vegetation (NV) in a headwater region of the Taihang Mountain. Water chemistry and N balances were quantified to estimate the accumulation and leaching of nitrate. Transport and fate of nitrate in the vadose zone were identified using stable isotopes of nitrate (δ18O-NO3-, δ15N-NO3-). Accumulation of NO3- was mainly in the shallow layer (0-50 cm) under both NV and FL in 2015. After large rain events (20-50 mm/day) during the rainy season in 2015, the leaching rates of NO3- at NV and FL sites were 86.04 and 9.61 kg/hm2, respectively. The accumulation of NO3- decreased 30% and 7% at NV and FL, respectively. However, the leaching rates of NO3- were 63.8 and 22.63 kg/hm2, and the accumulation of NO3- decreased 84% and 43% at NV and FL after extreme precipitation in 2016, respectively. Additionally, nitrate isotopes indicated that the different sources of nitrate in soil water were due to the land-use. However, nitrate isotopes showed that increased nitrate concentrations in groundwater at two sites in 2016 were due to the extreme precipitation. Accumulated high nitrate in surface soil in a normal year and leaching after extreme precipitation lead to increased nitrate concentration in groundwater, which poses a major threat in the future. The results are critical for informing land-use and water management when mitigating groundwater contamination in hilly area.

[Simulation of the Migration and Leaching of Nitrate Nitrogen in the Farmland Soil Profile in a Hilly Area of Taihang Mountain with the RZWQM Model].

Taihang Mountain region is the recharge area for groundwater in the North China Plain (NCP). In recent years, the elevated nitrate concentration in the groundwater in the Taihang Mountain has often been associated with the increased area of farmland and the excessive application of nitrogen. Thus, it is significant to study the soil nitrogen leaching process in typical farmland. In this study, the root zone water quality model (RZWQM) was used to simulate the nitrate nitrogen leaching of winter wheat/summer maize rotation systems in the Hilly Ecosystem Experimental Station in Taihang Mountain. The results showed that during the 2015-2016 winter wheat/summer maize season, the nitrate nitrogen from the soil leaching occurred mainly in the summer maize season (rainy season), with the nitrate nitrogen leaching amount reaching 59.9 kg·hm-2, while the nitrate nitrogen leaching amount during the winter wheat season was only 2.12 kg·hm-2. The soil nitrate nitrogen leaching condition was simulated using the RZWQM model for different nitrogen contents and years with different rainfall. Significant linear correlations were observed between nitrogen use and nitrate leaching in winter wheat/summer maize rotation systems. In summary, the results showed that the nitrate nitrogen leaching values were 10.5, 59.9, and 136.5 kg·hm-2 for nitrogen fertilizer applications of 0, 300, and 450 kg·hm-2, respectively, during extreme precipitation in a wet year (2015). The value of nitrate nitrogen leaching in the maize season of 2013 (dry year), 2015 (normal year), and 2016 (wet year) accounted for 9%, 10%, and 20% for the 300 kg·hm-2 of nitrogen fertilizer applied, respectively. However, the value of nitrate nitrogen leaching in the maize season of 2013 (dry year), 2015 (normal year), and 2016 (wet year) accounted for 11%, 17% and 30% of the 450 kg·hm-2 of nitrogen fertilizer applied. These results show that extreme precipitation events not only greatly recharge the groundwater, but also increase the leaching of accumulated nitrate nitrogen and potential nitrate contamination in the groundwater.

Ground water sustainability: methodology and application to the North China Plain.

This article analyzes part of a ground water flow system in the North China Plain (NCP) subject to severe overexploitation and rapid depletion. A transient ground water flow model was constructed and calibrated to quantify the changes in the flow system since the predevelopment 1950s. The flow model was then used in conjunction with an optimization code to determine optimal pumping schemes that improve ground water management practices. Finally, two management scenarios, namely, urbanization and the South-to-North Water Transfer Project, were evaluated for their potential impacts on the ground water resources in the study area. Although this study focuses on the NCP, it illustrates a general modeling framework for analyzing the sustainability, or the lack thereof, of ground water flow systems driven by similar hydrogeologic and economic conditions. The numerical simulation is capable of quantifying the various components of the overall flow budget and evaluating the impacts of different management scenarios. The optimization modeling allows the determination of the maximum "sustainable pumping" that satisfies a series of prescribed constraints. It can also be used to minimize the economic costs associated with ground water development and management. Furthermore, since the NCP is one of the most water scarce and economically active regions in the world, the conclusions and insights from this study are of general interest and international significance.

An improved straight-line fitting method for analyzing pumping test recovery data.

Theis (1935) derived an exact solution for the residual drawdown in a well after the cessation of a pumping test by summing two drawdowns: one (s1), caused by imaginary continuation of the original pumping and the other (s2), due to an imaginary injection at the same constant rate. We approximated the Theis solution to obtain a simple linear relation for determining the transmissivity and storage coefficient from recovery data. Unlike other existing straight-line fitting methods, in our method, we applied different approximations to the well functions in the solutions of s1 and s2. We used the well-known Cooper-Jacob approximation for s1, truncating the expansion of the well function in s2 to its first three terms. For the same level of truncation errors, while the Cooper-Jacob approximation requires the argument u1