Flavonoid 3-O-glucosyltransferase (3GT) is a key enzyme in the glucosidation of anthocyanins. To investigate the 3GT gene in rhododendron, we cloned an open reading frame (ORF) of 3GT gene (named Rh3GT) from Rhododendron hybridum Hort (Red cultivar) and then characterized this gene and the deduced protein in terms of the biochemical characteristics, expression level, and enzymatic function. The results showed that Rh3GT had a full length of 993 bp and encoded 330 amino acid residues. The deduced protein was hydrophilic, stable, weak acid, belonging to the glycosyltransferase family (GT-B type), with glutamine (Q) at position 44 in the PSPG box. The phylogenetic analysis showed that Rh3GT was most closely related to Vc3GT from Vaccinium corymbosum and Vm3GT from Vaccinium myrtillus. Rh3GT was expressed in the stems, leaves, and flowers and almost not expressed in the roots, with the highest expression level in petals during full blooming stage. Introduction of pCAMBIAL1302-Rh3GT into petals significantly up-regulated the expression level of Rh3GT and increased the total anthocyanin accumulation. Rh3GT was successfully expressed in Escherichia coli BL21 in the form of inclusion bodies with a size of about 36 kDa. The results of HPLC showed that the recombinant Rh3GT after denaturation, purification, and dilution could catalyze the synthesis of cyanidin and UDP-glucose to synthesize cyanidin 3-O-glucoside, indicating that the expressed protein had 3GT activity. This study provides basic data for further studying the molecular regulation mechanism of anthocyanin biosynthesis and theoretical support for molecular breeding of rhododendron.
Rhododendron/classification* ; Glucosyltransferases/metabolism* ; Cloning, Molecular ; Escherichia coli/metabolism* ; Recombinant Proteins/biosynthesis* ; Anthocyanins/biosynthesis* ; Phylogeny ; Plant Proteins/metabolism* ; Amino Acid Sequence

Objective To explore the therapeutic effect of traditional Chinese medicine(TCM)on subsolid nodule(SSN).Methods A practical randomized controlled study method,including 254 SSN patients was adopted.The patients were divided into the TCM(102 cases)and follow-up(152 cases)groups.The follow-up group received regular check-ups in accordance with the guidelines,and the TCM group received TCM syndrome differentiation treatment for 24 weeks.The two groups were compared in terms of the changes in their SSN diameter,SSN number,TCM symptom score,and overall therapeutic effect before and after treatment.Adverse reactions and safety indicators were also recorded.Results The TCM group showed a significantly higher effective rate of treatment(16.7%)than the follow-up group(2.6%)(P<0.01).Compared with their condition before treatment,the TCM group showed no significant changes in their SSN diameter and number but presented considerably reduced fatigue,yellow and red urine symptoms,and overall TCM symptom score(P<0.05).The follow-up group exhibited significantly increased diameter and number of SSN(P<0.01).The follow-up group showed the significantly higher increase in SSN diameter after treatment than the TCM group(P<0.05).Moreover,the follow-up group showed significantly higher fatigue,depression,yellow and red urine symptom scores,and overall TCM symptom score than the TCM group(P<0.05 or P<0.01).Conclusion TCM treatment for SSN has a distinct clinical efficacy,reduces the malignant risk of SSN and improves clinical symptoms of SSN patients,and is safe and feasible.

Objective:To assess the altered right atrial(RA) function using two-dimensional speckle-tracking imaging(2D-STI) in patients with persistent pulmonary hypertension of the newborn(PPHN), and investigate the diagnostic value of different right heart strain parameters in PPHN.Methods:Ultrasound images of 42 newborns with clinically confirmed diagnosis of PPHN in the neonatal intensive care unit of Shenzhen Children′s Hospital (median gestational age 37 + 1 weeks, median age 6 d) and 30 normal newborns (control group, median gestational age 37 + 3 weeks, median age 6.5 d) from January 2020 to January 2023 were retrospectively analyzed, all with gestational age ≥34 gestational weeks. 2D-STI was applied to evaluate RA function: RA strain(RA LS) and area change fraction(RA FAC), where RA LS included RA reservoir strain (εs), RA conduit strain (εe) and RA active contract strain (εa), while evaluating right ventricle(RV) function: RV global longitudinal strain(RV GLS), RV FAC, etc. And the degree of pulmonary artery pressure(PAP) was assessed by tricuspid regurgitation velocity(TRV). The above parameters were compared between the PPHN group and the control group, and the correlations between RA function parameters, RV function parameters and PAP in PPHN group were analyzed. The area under ROC curve (AUC) was used to compare the accuracy of each parameter in the evaluation of impaired cardiac function in PPHN patients. Results:Compared with the control group, RA function (εs, εe, εa and RA FAC) were impaired in PPHN patients (all P<0.05). εs was positively correlated with RV GLS, RA FAC ( r=0.494, 0.356, both P<0.05) and negatively correlated with minimum right atrial area (RAA min), pulmonary artery diameter (PAD), and tricuspid annular internal diameter ( rs=-0.285, r=-0.495, -0.396; both P<0.05); εe was negatively correlated with PAD ( rs=-0.256, P<0.05); εa was positively correlated with RV GLS ( r=0.499, P<0.05) and negatively correlated with PAD and tricuspid annular internal diameter ( r=-0.390, -0.380; both P<0.05); RA FAC was positively correlated with RV GLS ( r=0.365, P<0.05) and negatively correlated with PAD and tricuspid annular internal diameter ( r=-0.439, -0.328; both P<0.05). RA LS and RA FAC had no correlations with TRV-estimated PAP ( P>0.05). ROC analysis showed that the sensitivity and specificity of εs<40.50% for diagnosing PPHN was 0.905 and 0.800, respectively, with an AUC of 0.929; the sensitivity and specificity of RV GLS<18.55% for diagnosing PPHN were 0.905 and 0.900, respectively, with an AUC of 0.963; εs combined with RV GLS was the best indicator for early detection of right heart functional impairment in PPHN, with sensitivity and specificity of 0.905 and 1.000, respectively, and AUC was 0.985. Conclusions:RA function and RV function are both impaired in PPHN patients. εs combined with RV GLS is the best indicator for early assessment of right heart function impairment and diagnosis of PPHN.