[Multiple precision behavioral therapy for mild to moderate stress urinary incontinence with sexual dysfunction in women].

OBJECTIVE: To explore the clinical effect of multiple precision behavioral therapy (MPBT) on mild to moderate stress urinary incontinence (SUI) with female sexual dysfunction (FSD) in women. METHODS: We randomly divided 90 female patients with mild to moderate SUI with FSD into three groups of an equal number: control group A, control group B and an MPBT group, treated by electrical stimulation, Kegel training and MPBT, respectively, all for 8 weeks. Using International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF), Incontinence Impact Questionnaire (IIQ-7), Female Sexual Function Indexes (FSFI) and Glazer protocol, we evaluated the clinical effects, recorded the cost of treatment, and compared them among the three groups of patients. RESULTS: Totally, 87 of the patients completed the treatment, 27 in control group A, 30 in control group B and 30 in the MPBT group. There was no significant difference in the baseline data among the three groups (P > 0.05). ICIQ-SF and IIQ-7 scores, FSFI and Glazer values were remarkably improved in the MPBT group after treatment (P < 0.05). The therapeutic effect was significantly better and the treatment cost markedly lower in the MPBT than in the control groups (P < 0.05). CONCLUSION: Multiple precision behavioral therapy can effectively improve the clinical symptoms of mild to moderate stress urinary incontinence and sexual dysfunction in women, with low cost and high safety.

[Effect of functional acupoint electrical stimulation combined with tadalafil on erectile dysfunction in middle-aged and elderly patients].

OBJECTIVE: To investigate the application effect of functional acupoint electrical stimulation combined with tadara irregular administration in middle-aged and elderly patients with erectile dysfunction (ED), and to provide reference for clinical treatment. METHODS: A total of 40 middle-aged and elderly patients with ED admitted to the pelvic floor Center of our hospital from March 2021 to March 2023 were randomly divided into two groups with 20 cases in each group.The control group was treated with tadalafil regularly, and the observation group was treated with functional acupoint electrical stimulation on the basis of this treatment. The total course of treatment was 6 weeks.The clinical efficacy of the two groups was compared. The therapeutic efficacy was evaluated by the International Erectile Function Index (IIEF-5), penile hardness score (EHS), serum total testosterone (TT) level, sexual satisfaction scale (SS) and pelvic floor electromyography, and the occurrence of adverse events was recorded. RESULTS: The total effective rate of the observation group was significantly higher than that of the control group (90% vs 70%, P < 0.05). After 6 weeks of treatment, both groups showed improvements in IIEF-5, EHS, SS, and TT compared to before treatment (P < 0.01). However, the improvement in the observation group was significantly better than that in the control groupï¼»IIEF-5: (22.13±2.11) vs (19.69±2.04), EHS: (3.68±0.47) vs (2.89±0.60), SS: (77.41±7.59) vs (70.32±7.28), TT: (13.43±3.89) nmol/L vs (8.85±3.02) nmol/L, all P < 0.01ï¼½; There were no significant changes in pelvic floor muscle electromyography values in the control group before and after treatment (P > 0.05), while in the observation group, pelvic floor muscle electromyography values (PFMV) in the pre-resting phase, fast muscle (Type II muscle) phase, slow muscle (Type I muscle) phase, endurance testing phase, and post-resting phase all improved compared to before treatment and were superior to the control group (P < 0.05). CONCLUSION: Functional acupoint electrical stimulation combined with tadara irregular administration can improve the therapeutic effect of middle-aged and elderly patients with ED, improve pelvic floor function, safe and reliable.

Regeneration and Agrobacterium-mediated transformation of the apomictic species Eulaliopsis binata.

Protocols for regeneration and Agrobacterium-mediated transformation of the apomictic species Eulaliopsis binata were developed. Initially, seeds of four genotypes of E. binata were incubated on a callus induction Murashige and Skoog (MS) basal medium supplemented with three concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D). It was found that 36.2 % of explants developed highly friable callus on medium containing 3.0 mg l(-1) 2,4-D. Based on frequency of callus induction, the genotype Neixiang was selected for regeneration and transformation. Callus incubated on MS basal medium supplemented with 0.2 mg l(-1) α-naphthalene acetic acid and 6.0 mg l(-1) 6-furfuryl-aminopurine developed shoots. Subsequently, Agrobacterium tumefaciens strain EHA105-harboring a plasmid pCAMBIA1381 carrying a hygromycin phosphotransferase (hpt) resistance gene and a synthetic green fluorescent protein (GFP) gene, both driven by the cauliflower mosaic virus 35S promoter-was used for transformation system. Putative transgenic callus was obtained following two cycles of hygromycin selection. Expression of the transgene(s) in putative transgenic callus was analyzed using the GFP detection. Molecular identification of putative transformed shoots was performed by polymerase chain reaction and Southern blot analysis to confirm presence and integration of the hpt gene.

PtSVP, an SVP homolog from trifoliate orange (Poncirus trifoliata L. Raf.), shows seasonal periodicity of meristem determination and affects flower development in transgenic Arabidopsis and tobacco plants.

A MADS-box gene was isolated using the suppressive subtractive hybridization library between early-flowering mutant and wild-type trifoliate orange (Poncirus trifoliata L. Raf.). This gene is highly homologous with Arabidopsis SHORT VEGETATIVE PHASE (SVP). Based on real-time PCR and in situ hybridization during bud differentiation, PtSVP was expressed intensively in dormant tissue and vegetative meristems. PtSVP transcripts were detected in apical meristems before floral transition, then down-regulated during the transition. PtSVP expression was higher in differentiated (flower primordium) than in undifferentiated cells (apical meristems). The PtSVP expression pattern during apical meristem determination suggested that its function is not to depress flower initiation but to maintain meristem development. Transcription of PtSVP in Arabidopsis svp-41 showed partially rescued SVP function. Ectopic overexpression of PtSVP in wild-type Arabidopsis induced late flowering similar to the phenotypes induced by other SVP/StMADS-11-like genes, but transformants produced additional trichomes and floral defects, such as flower-like structures instead of carpels. Ectopic expression of PtSVP in tobacco also caused additional florets. Overexpression of PtSVP in tobacco inhibited early transition of the coflorescence and prolonged coflorescence development, thus causing additional florets at the later stage. A yeast two-hybrid assay indicated that PtSVP significantly interacted with PtAP1, a homolog of Arabidopsis APETALA1 (AP1). These findings suggest that citrus SVP homolog genes are involved in flowering time regulation and may influence inflorescence meristem identity in some conditions or genetic backgrounds. SVP homologs might have evolved among plant species, but the protein functions are conserved between Arabidopsis and citrus.

Tetraploidization of diploid Dioscorea results in activation of the antioxidant defense system and increased heat tolerance.

Polyploidy is reported to show increased tolerance to environmental stress. In this work, tetraploid plants of Dioscorea zingiberensis were obtained by colchicine treatment of shoots propagated in vitro. The highest tetraploid induction rate was achieved by treatment with 0.15% colchicine for 24h. Diploid and tetraploid plants were exposed to normal (28 degrees C) and high temperature (42 degrees C) for 5d during which physiological indices were measured. Compared with diploid plants, relative electrolyte leakage and contents of malondialdehyde, superoxide anions and hydrogen peroxide were lower in tetraploids, while activities of antioxidant enzymes, such as superoxide dismutase, peroxidase, catalase, ascorbate peroxidase and glutathione reductase, were stimulated and antioxidants (ascorbic acid and glutathione) were maintained at high concentrations. These results indicate that tetraploid plants possess a stronger antioxidant defense system and increased heat tolerance.

PtFLC homolog from trifoliate orange (Poncirus trifoliata) is regulated by alternative splicing and experiences seasonal fluctuation in expression level.

In many plant species, exposure to a prolonged period of low temperature during the winter promotes flowering in the spring, a process termed vernalization. In Arabidopsis, the vernalization requirement of winter annual ecotypes is caused by a MADS-box gene FLOWERING LOCUS C (FLC), which is a repressor of flowering gene. Here, a MADS-box gene was isolated from an early flowering trifoliate orange mutant (precocious trifoliate orange, Poncirus trifoliata L. Raf) by the RACE method combined with a cDNA library. Phylogenetic analysis reveals that the MADS-box gene is more closely related to the homologs of the FLOWERING LOCUS C lineage than to any of the other MIKC-type MADS-box lineages known from Arabidopsis. The expression profile of the MADS-box gene by real-time PCR showed upregulation of PtFLC during the winter, followed by a decrease in the spring and summer. This kind of cycling is contrary to the pattern observed in Arabidopsis. In situ hybridization reveals that the MADS-box gene is predominately expressed in the vegetative and reproductive meristems. In addition, five alternatively spliced transcripts of the MADS-box gene were also isolated at juvenile and adult mutant developmental stages. Expression analysis of these transcripts at different developmental stages indicated involvement of alternative splicing during phase change. The information suggests a complicated regulation mechanism in seasonal response and flower formation in perennial woody plants.

Identification of flowering-related genes between early flowering trifoliate orange mutant and wild-type trifoliate orange (Poncirus trifoliata L. Raf.) by suppression subtraction hybridization (SSH) and macroarray.

To gain a better understanding of gene expression in early flowering trifoliate orange mutant (precocious trifoliate orange, Poncirus trifoliata L. Raf.), we performed suppression subtractive hybridization, which allowed identification of flowering-related genes in the mutant and the wild type in the juvenile phase. Using macroarray analysis, we identified 125 and 149 non-redundant expressed sequence tags (ESTs) in the forward-subtracted and the reverse-subtracted library. These cDNAs covered a broad repertoire of flowering development related genes, provided helpful information for understanding genetic mechanism underlying the signaling and regulation in transition from the vegetative to reproductive phase. We have investigated the temporal and spatial expression pattern of some SSH-enriched flowering-related genes in the mutant and the wild type. Of these genes, three genes (BARELY ANY MERITED, FLOWERING LOCUS T and TERMINAL FLOWER1) encoding proteins previously reported to be associated with, or involved in, developmental processes in other species were identified and further investigated by in situ hybridization. Specific spatial and/or temporal patterns were detected, and differences were observed between the mutant and the wild type during flower development. Meanwhile, the temporal expression of these genes was further examined by real-time PCR, the results showed that FT and BAM transcripts accumulated to higher levels and TFL1 transcripts accumulated to lower levels in mutant juvenile tissues relative to wild-type juvenile tissues. In the adult stage, FT, BAM and TFL1 expression patterns were closely correlated with flowering development, suggesting that these three genes may play a critical role in the early flowering process of precocious trifoliate orange.

Identification of early-flower-related ESTs in an early-flowering mutant of trifoliate orange (Poncirus trifoliata) by suppression subtractive hybridization and macroarray analysis.

To increase our understanding of the genes involved in flower development in citrus, we identified genes differentially expressed between juveniles and adults of an early-flowering mutant of trifoliate orange (precocious trifoliate orange, Poncirus trifoliata L. Raf.) by suppressive subtractive hybridization (SSH) and macroarray hybridization analyses. Based on dot blot analysis, we confirmed that the juvenile subtracted cDNA library comprised genes upregulated during floral induction, inflorescence development and flowering. A total of 190 nonredundant expressed sequence tags were identified that were related flower development. The temporal and spatial expression patterns of four SSH-enriched genes were studied in adult precocious trifoliate orange by real-time PCR. Among these differentially expressed genes, the expression pattern of C5-B16 was closely correlated with inflorescence development and flowering. The full-length cDNA of C5-B16 was isolated by 5' and 3' rapid amplification of cDNA ends. Sequence analysis indicated that C5-B16 is a novel gene in citrus.