Clinical pharmacists' involvement in carbapenem antibiotics management at Wenzhou Integrated Hospital.

BACKGROUND: Carbapenem antibiotics are a pivotal solution for severe infections, particularly in hospital settings. The emergence of carbapenem-resistant bacteria owing to the irrational and extensive use of carbapenems underscores the need for meticulous management and rational use. Clinical pharmacists, with their specialized training and extensive knowledge, play a substantial role in ensuring the judicious use of carbapenem. This study aimed to elucidate the patterns of carbapenem use and shed light on the integral role played by clinical pharmacists in managing and promoting the rational use of carbapenem antibiotics at Wenzhou Integrated Traditional Chinese and Western Medicine Hospital. AIM: To analyze carbapenem use patterns in our hospital and role of clinical pharmacists in managing and promoting their rational use. METHODS: We performed a retrospective analysis of carbapenem use at our hospital between January 2019 and December 2021. Several key indicators, including the drug utilization index, defined daily doses (DDDs), proportion of antimicrobial drug costs to total hospitalization expenses, antibiotic utilization density, and utilization rates in different clinical departments were comprehensively analyzed. RESULTS: Between 2019 and 2021, there was a consistent decline in the consumption and sales of imipenem-cilastatin sodium, meropenem (0.3 g), and meropenem (0.5 g). Conversely, the DDDs of imipenem-cilastatin sodium for injection increased in 2020 and 2021 vs 2019, with a B/A value of 0.67, indicating a relatively higher drug cost. The DDDs of meropenem for injection (0.3 g) exhibited an overall upward trend, indicating an increasing clinical preference. However, the B/A values for 2020 and 2021 were both > 1, suggesting a relatively lower drug cost. The DDDs of meropenem for injection (0.5 g) demonstrated a progressive increase annually and consistently ranked first, indicating a high clinical preference with a B/A value of 1, signifying good alignment between economic and social benefits. CONCLUSION: Carbapenem use in our hospital was generally reasonable with a downward trend in consumption and sales over time. Clinical pharmacists play a pivotal role in promoting appropriate use of carbapenems.

Investigation of the Inhibitory Effect of Simvastatin on the Metabolism of Lidocaine Both in vitro and in vivo.

BACKGROUND: Lidocaine has cardiovascular and neurologic toxicity, which is dose-dependent. Due to CYP3A4-involved metabolism, lidocaine may be prone to drug-drug interactions. MATERIALS AND METHODS: Given statins have the possibility of combination with lidocaine in the clinic, we established in vitro models to assess the effect of statins on the metabolism of lidocaine. Further pharmacokinetic alterations of lidocaine and its main metabolite, monoethylglycinexylidide in rats influenced by simvastatin, were investigated. RESULTS: In vitro study revealed that simvastatin, among the statins, had the most significant inhibitory effect on lidocaine metabolism with IC50 of 39.31 µM, 50 µM and 15.77 µM for RLM, HLM and CYP3A4.1, respectively. Consistent with in vitro results, lidocaine concomitantly used with simvastatin in rats was associated with 1.2-fold AUC(0-t), 1.2-fold AUC(0-∞), and 20%-decreased clearance for lidocaine, and 1.4-fold Cmax for MEGX compared with lidocaine alone. CONCLUSION: Collectively, these results implied that simvastatin could evidently inhibit the metabolism of lidocaine both in vivo and in vitro. Accordingly, more attention and necessary therapeutic drug monitoring should be paid to patients with the concomitant coadministration of lidocaine and simvastatin so as to avoid unexpected toxicity.

Natural variation in the promoter of OsHMA3 contributes to differential grain cadmium accumulation between Indica and Japonica rice.

Rice is a major source of cadmium (Cd) intake for Asian people. Indica rice usually accumulates more Cd in shoots and grains than Japonica rice. However, underlying genetic bases for differential Cd accumulation between Indica and Japonica rice are still unknown. In this study, we cloned a quantitative trait locus (QTL) grain Cd concentration on chromosome 7 (GCC7) responsible for differential grain Cd accumulation between two rice varieties by performing QTL analysis and map-based cloning. We found that the two GCC7 alleles, GCC7PA64s and GCC793-11 , had different promoter activity of OsHMA3, leading to different OsHMA3 expression and different shoot and grain Cd concentrations. By analyzing the distribution of different haplotypes of GCC7 among diverse rice accessions, we discovered that the high and low Cd accumulation alleles, namely GCC793-11 and GCC7PA64s , were preferentially distributed in Indica and Japonica rice, respectively. We further showed that the GCC7PA64s allele can be used to replace the GCC793-11 allele in the super cultivar 93-11 to reduce grain Cd concentration without adverse effect on agronomic traits. Our results thus reveal that the QTL GCC7 with sequence variation in the OsHMA3 promoter is an important determinant controlling differential grain Cd accumulation between Indica and Japonica rice.

A defensin-like protein drives cadmium efflux and allocation in rice.

Pollution by heavy metals limits the area of land available for cultivation of food crops. A potential solution to this problem might lie in the molecular breeding of food crops for phytoremediation that accumulate toxic metals in straw while producing safe and nutritious grains. Here, we identify a rice quantitative trait locus we name cadmium (Cd) accumulation in leaf 1 (CAL1), which encodes a defensin-like protein. CAL1 is expressed preferentially in root exodermis and xylem parenchyma cells. We provide evidence that CAL1 acts by chelating Cd in the cytosol and facilitating Cd secretion to extracellular spaces, hence lowering cytosolic Cd concentration while driving long-distance Cd transport via xylem vessels. CAL1 does not appear to affect Cd accumulation in rice grains or the accumulation of other essential metals, thus providing an efficient molecular tool to breed dual-function rice varieties that produce safe grains while remediating paddy soils.

Inhibitory Effect of Apigenin on Losartan Metabolism and CYP2C9 Activity in vitro.

BACKGROUND: CYP2C9 is one of the most important phase I drug-metabolizing enzymes in liver. The objective of this work was to investigate the effects of apigenin on the metabolism of losartan and human CYP2C9 and rat CYP2C11 activity in vitro. METHODS: Different concentrations of apigenin were added to a 100 mmol/l Tris-HCl reaction mixture containing 2 pmol/ml recombinant human CYP2C9.1, 0.25 mg/ml human liver microsomes or 0.5 mg/ml rat liver microsomes to determine the half maximal inhibition or a half-maximal inhibitory concentration (IC50) on the metabolism of losartan. In addition, diclofenac used as CYP2C9 substrate was performed to determine the effects of apigenin on CYP2C9. RESULTS: The results showed that apigenin has the inhibitory effect on the metabolism of losartan in vitro, the IC50 was 7.61, 4.10 and 11.07 µmol/l on recombinant CYP2C9 microsomes, human liver microsomes and rat liver microsomes, respectively. Meanwhile, apigenin's mode of action on human CYP2C9 activity was competitive for the substrate diclofenac. In contrast to its potent inhibition of CYP2C9 in humans (9.51 µmol/l), apigenin had lesser effects on CYP2C11 in rat (IC50 = 15.51 µmol/l). CONCLUSION: The observations imply that apigenin has the inhibitory effect on the metabolism of losartan and CYP2C9 activity in vitro. More attention should be paid as to when losartan should be administrated combined with apigenin.

LSCHL4 from Japonica Cultivar, which is allelic to NAL1, increases yield of indica super rice 93-11.

The basic premise of high yield in rice is to improve leaf photosynthetic efficiency and coordinate the source-sink relationship in rice plants. Quantitative trait loci (QTLs) related to morphological traits and chlorophyll content of rice leaves were detected at the stages of heading to maturity, and a major QTL (qLSCHL4) related to flag leaf shape and chlorophyll content was detected at both stages in recombinant inbred lines constructed using the indica rice cultivar 93-11 and the japonica rice cultivar Nipponbare. Map-based cloning and expression analysis showed that LSCHL4 is allelic to NAL1, a gene previously reported in narrow leaf mutant of rice. Overexpression lines transformed with vector carrying LSCHL4 from Nipponbare and a near-isogenic line of 93-11 (NIL-9311) had significantly increased leaf chlorophyll content, enlarged flag leaf size, and improved panicle type. The average yield of NIL-9311 was 18.70% higher than that of 93-11. These results indicate that LSCHL4 had a pleiotropic function. Exploring and pyramiding more high-yield alleles resembling LSCHL4 for super rice breeding provides an effective way to achieve new breakthroughs in raising rice yield and generate new ideas for solving the problem of global food safety.

Dissecting yield-associated loci in super hybrid rice by resequencing recombinant inbred lines and improving parental genome sequences.

The growing world population and shrinkage of arable land demand yield improvement of rice, one of the most important staple crops. To elucidate the genetic basis of yield and uncover its associated loci in rice, we resequenced the core recombinant inbred lines of Liang-You-Pei-Jiu, the widely cultivated super hybrid rice, and constructed a high-resolution linkage map. We detected 43 yield-associated quantitative trait loci, of which 20 are unique. Based on the high-density physical map, the genome sequences of paternal variety 93-11 and maternal cultivar PA64s of Liang-You-Pei-Jiu were significantly improved. The large recombinant inbred line population combined with plentiful high-quality single nucleotide polymorphisms and insertions/deletions between parental genomes allowed us to fine-map two quantitative trait loci, qSN8 and qSPB1, and to identify days to heading8 and lax panicle1 as candidate genes, respectively. The quantitative trait locus qSN8 was further confirmed to be days to heading8 by a complementation test. Our study provided an ideal platform for molecular breeding by targeting and dissecting yield-associated loci in rice.

Characterization of physiological response and identification of associated genes under heat stress in rice seedlings.

Global warming, which is caused by greenhouse gas emissions, makes food crops more vulnerable to heat stress. Understanding the heat stress-related mechanisms in crops and classifying heat stress-related genes can increase our knowledge in heat-resistant molecular biology and propel developments in molecular design breeding, which can help rice cope with unfavorable temperatures. In this study, we carried out a physiological analysis of rice plants after heat stress. The results show a dramatic increase in malondialdehyde contents and SOD activities. We successfully isolated 11 heat-related rice genes with known function annotation through DNSH, which is an improved SSH method for screening long cDNA fragments. The reanalysis of microarray data from public database revealed that all these genes displayed various expression patterns after heat stress, drought, cold and salt. Quantitative real-time reverse transcription PCR was also performed to validate the expression of these genes after heat stress. The expressions in 10 genes were all significantly changed except for contig 77, which is a CBL-interacting protein kinase. Several reports have been published about the members of the same gene family.

Mapping quantitative trait loci associated with arsenic accumulation in rice (Oryza sativa).

The quantitative trait loci (QTLs) associated with arsenic (As) accumulation in rice were mapped using a doubled haploid population established by anther culture of F1 plants from a cross between a Japonica cultivar CJ06 and an Indica cultivar TN1 (Oryza sativa). Four QTLs for arsenic (As) concentrations were detected in the map. At the seedling stage, one QTL was mapped on chromosome 2 for As concentrations in shoots with 24.4% phenotypic variance and one QTL for As concentrations in roots was detected on chromosome 3. At maturity, two QTLs for As concentrations in grains were found on chromosomes 6 and 8, with 26.3 and 35.2% phenotypic variance, respectively. No common loci were detected among these three traits. Interestingly, the QTL on chromosome 8 was found to be colocated for As concentrations in grain at maturity and shoot phosphorus (P) concentrations at seedling stage. These results provide an insight into the genetic basis of As uptake and accumulation in rice, and will be useful in identifying genes associated with As accumulation.

[Genetic analysis of two extremely segregation distorted populations in rice (Oryza sativa L.)].

Segregation distortion is a quite common phenomenon in living species and thought to be a potent evolutional force. The main reasons of distorted segregation ratios are responsible for the selection of gametes or sporophytes. In this study, two extreme segregation distortions from the progenies of lmi x 02428 and d6 x 93-11 were identified. The segregation ratio of molecular markers tightly linked with LMI and D6 genes were analyzed and skew segregation were found in the markers tested which were indicated by significant deviation from the expected Mendelian segregation ratio(1:2:1). The segregation distorted regions were detected between molecular markers ST8 and ST8-2 near the centromere of chromosome 8, and ST7-1 and ST7-3 near telomere of chromosome 7, respectively. Meanwhile, the results indicated that segregation distortion had related with the different crossed combinations.

A large-scale field study of transgene flow from cultivated rice (Oryza sativa) to common wild rice (O. rufipogon) and barnyard grass (Echinochloa crusgalli).

The introgression of transgenes into wild relatives or weeds through pollen-mediated gene flow is a major concern in environmental risk assessment of transgenic crops. A large-scale (1.3-1.8 ha) rice gene flow study was conducted using transgenic rice containing the bar gene as a pollen donor and Oryza rufipogon as a recipient. There was a high frequency of transgene flow (11%-18%) at 0-1 m, with a steep decline with increasing distance to a detection limit of 0.01% by 250 m. To our knowledge, this is the highest frequency and longest distance of gene flow from transgenic rice to O. rufipogon reported so far. On the basis of these data, an adequate isolation distance from both conventional and transgenic rice should be taken for in situ conservation of common wild rice. Meanwhile, there is no evidence of transgene introgression into barnyard grass, even when it has coexisted with transgenic rice containing the bar gene for five successive years. Thus, the environmental risk of gene flow to this weedy species is of little concern.

Genetic dissection for leaf correlative traits of rice (Oryza sativa L.) under drought stress.

Water is becoming a restricted factor of agricultural development owing to the global shortage of water resources. Screening and improving drought tolerant rice cultivars would be helpful for increasing and stabilizing yield, economizing water and reducing environmental pollution. In this study, 127 rice lines of DII population derived from an indica variety Zhaiyeqing 8 (ZYQ8) and a japonica variety Jingxi 17 (JX17) were used to locate QTLs for leaf rolling, relative water content and rate of electric conductivity under drought stress. The results showed that significant differences between the parents were detected for all measured traits. The tremendous transgressive segregations for these traits were observed in the population. The frequency of all traits in the population was approximately normally distributed with slight skew. A total of six QTLs for the three traits were detected with molecular linkage map of 234 markers, including three QTLs (qLR-1, qLR-5 and qLR-11) for leaf rolling, two QTLs (qRWC-1 and qRWC-6) for relative water content and one QTL (qREC-6) for rate of electric conductivity. Visual measurement for leaf rolling can be used to screen a large number of rice germplasm resources or varieties, which is of importance to screening and utilization of drought tolerant rice varieties.

[Genetic dissection of cooked rice elongation in rice (Oryza sativa L.)].

QTLs for milled rice length (MRL), cooked rice length (CRL) and cooked rice elongation (CRE) were identified by using a population of 127 DH lines derived from a cross between ZYQ8 and JX17. Totally, 14 QTLs for rice elongation traits were detected on chromosomes 1, 2, 3, 5, 6, 7, 10, 11 and 12. Two putative QTLs for MRL were mapped on chromosomes 2. Seven putative QTLs for CRL were mapped on chromosomes 1, 6, 7, 10, 11 and 12. Five putative QTLs for CRE were mapped on chromosomes 5, 6, and 10. The regions of G249-G164 on chromosome 3, G30-RZ516 on chromosome 6 and G1082-GA223 on chromosome 10 were detected simultaneously for affecting cooked rice length and cooke rice elongation. LODs of the QTLs related to rice elongation varied from 2.26 to 9.25, and their explained variations from 5.31% to 17.21% . It is indicated that cooked rice elongation was controlled by polygene and Wx-gene located on the same region with qCRE-6 was important to cooked rice elongation.

[Analysis of gene loci and epistasis for drought tolerance in seedling stage of rice (Oryza sativa L.)].

Drought tolerance of rice is important because a considerable proportion of the world rice area is not irrigated and is prone to water deficit. In this study, an indica variety, Zhai Ye Qing 8 (ZYQ8), and a japonica variety, Jing Xi 17 (JX17), and their double haploid (DH) population were used for genetic study of drought tolerance. Water supply was stopped in seedling period for 15 days and then drought tolerance of the DH population and their parents were investigated. Mapping quantitative trait loci (QTLs) was undertaken base on the constructed molecular linkage map of this population. Two QTLs (qDR-5 and qDR-12) for drought tolerance were identified, they were in the region of GA41-GA257 on chromosome 5 and RG457-Y12817R on chromosome 12, respectively. The tolerance alleles of both QTLs were from the indica parent, ZYQ8. In the meantime two genes for drought tolerance near GA257 and Y12817R were detected too by using Epistat software, that is in accordance with the result by using Mapmaker/QTL. In addition, three loci (RG541, G318 and G192 on chromosome 1, 4 and 8, respectively) were found interacting with GA257 by Epistat software, while one locus (CT234 on chromosome 3) found interacting with Y12817R were also detected by Epistat software.

[QTL analysis of sheath blight resistance in rice (Oryza sativa L.)].

In this study, an indica variety Zhai Ye Qing 8 (ZYQ8) and a japonica variety Jing Xi 17(JX17) and their double haploid (DH) population were used. Sheath blight resistance of the DH population and their parents was investigated using a syringe inoculation method. Quantitative trait loci (QTL) analysis was undertaken based on a constructed molecular linkage map of this population in Hangzhou and Hainandao respectively. A total of four sheath blight resistant QTLs (qSBR-2, qSBR-3, qSBR-7 and qSBR-11) were identified, and located on chromosome 2, 3, 7 and 11, respectively. Resistant alleles of qSBR-2, qSBR-3, and qSBR-7 were derived from the resistant parent ZYQ8, whereas resistant allele of qSBR-11 from susceptible parent JX17. qSBR-2, qSBR-3 and qSBR-7 were detected both in Hangzhou and Hainandao, whereas qSBR-11 was detected only in Hangzhou. Sheath blight response was significantly correlated with culm length and heading date in Hangzhou's test. Among the QTLs for culm length and heading date, only qCL-3 for culm length, was in the same chromosome region of qSBR-3, the other QTLs were not linked to the QTLs for sheath blight resistance.