Association between polypharmacy and 2-year outcomes among Chinese older inpatients: a multi-center cohort study.

BACKGROUND: The escalating global prevalence of polypharmacy presents a growing challenge to public health. In light of this issue, the primary objective of our study was to investigate the status of polypharmacy and its association with clinical outcomes in a large sample of hospitalized older patients aged 65 years and over. METHODS: A two-year prospective cohort study was carried out at six tertiary-level hospitals in China. Polypharmacy was defined as the prescription of 5 or more different medications daily, including over-the-counter and non-prescription medications. Baseline polypharmacy, multimorbidity, and other variables were collected when at admission, and 2-year outcomes were recorded by telephone follow-up. We used multivariate logistic regression analysis to examine the associations between polypharmacy and 2-year outcomes. RESULTS: The overall response rate was 87.2% and 8713 participants were included in the final analysis. The mean age was 72.40 years (SD = 5.72), and women accounted for 42.2%. The prevalence of polypharmacy among older Chinese inpatients is 23.6%. After adjusting for age, sex, education, marriage status, body mass index, baseline frailty, handgrip strength, cognitive impairment, and the Charlson comorbidity index, polypharmacy is significantly associated with frailty aggravation (OR 1.432, 95% CI 1.258-1.631) and mortality (OR 1.365, 95% CI 1.174-1.592), while inversely associated with readmission (OR 0.870, 95% CI 0.764-0.989). Polypharmacy was associated with a 35.6% increase in the risk of falls (1.356, 95%CI 1.064-1.716). This association weakened after adjustment for multimorbidity to 27.3% (OR 1.273, 95%CI 0.992-1.622). CONCLUSIONS: Polypharmacy was prevalent among older inpatients and was a risk factor for 2-year frailty aggravation and mortality. These results highlight the importance of optimizing medication use in older adults to minimize the risks associated with polypharmacy. Further research and implementing strategies are warranted to enhance the quality of care and safety for older individuals exposed to polypharmacy. TRIAL REGISTRATION: Chinese Clinical Trial Registry, ChiCTR1800017682, registered 09/08/2018.

Identification and profiling of upland cotton microRNAs at fiber initiation stage under exogenous IAA application.

BACKGROUND: Cotton is the most essential textile crop worldwide, and phytohormones are critical for cotton fiber development. One example is the role of auxin in fiber initiation, but we know little molecular basis. MicroRNAs (miRNAs) have a significant function in cotton development; nevertheless their role in fiber initiation remains unclear. Here, exogenous IAA was applied to cotton plant before anthesis. Utilizing small RNA sequencing, the mechanism underlying miRNA-mediated regulation of fiber initiation under exogenous IAA treatment was investigated. RESULTS: With exogenous IAA application, the endogenous IAA and GA contents of IAA treated (IT) ovules were higher than control (CK) ovules at the fiber initiation stage, while endogenous ABA content was lower in IT than CK. Using scanning electron microscopy, we found the fiber number and size were significantly promoted in IT at 0 DPA. Fiber quality analysis showed that fiber length, uniformity, strength, elongation, and micronaire of IT were higher than CK, though not statistically significant, while lint percent was significantly higher in IT. We generated six small RNA libraries using - 3, 0, and 3 DPA ovules of IT and CK, and identified 58 known miRNAs and 83 novel miRNAs together with the target genes. The differential expressed miRNAs number between IT and CK at - 3, 0, 3 DPA was 34, 16 and 24, respectively. Gene ontology and KEGG pathway enrichment analyses for the target genes of the miRNAs expressed in a differential manner showed that they were significantly enriched in 30 terms and 8 pathways. QRT-PCR for those identified miRNAs and the target genes related to phytohormones and fiber development was performed, and results suggested a potential role of these miRNAs in fiber initiation. CONCLUSIONS: The exogenous IAA application affected the relative phytohormone contents in ovule and promoted fiber initiation in cotton. Identification and profiling of miRNAs and their targets at the fiber initiation stage provided insights for miRNAs' regulation function of fiber initiation. These findings not only shed light on the regulatory network of fiber growth but also offer clues for cotton fiber amelioration strategies in cotton.

The Ca2+ channel beta subunit determines whether stimulation of Gq-coupled receptors enhances or inhibits N current.

In superior cervical ganglion (SCG) neurons, stimulation of M(1) receptors (M(1)Rs) produces a distinct pattern of modulation of N-type calcium (N-) channel activity, enhancing currents elicited with negative test potentials and inhibiting currents elicited with positive test potentials. Exogenously applied arachidonic acid (AA) reproduces this profile of modulation, suggesting AA functions as a downstream messenger of M(1)Rs. In addition, techniques that diminish AA's concentration during M(1)R stimulation minimize N-current modulation. However, other studies suggest depletion of phosphatidylinositol-4,5-bisphosphate during M(1)R stimulation suffices to elicit modulation. In this study, we used an expression system to examine the physiological mechanisms regulating modulation. We found the beta subunit (Ca(V)beta) acts as a molecular switch regulating whether modulation results in enhancement or inhibition. In human embryonic kidney 293 cells, stimulation of M(1)Rs or neurokinin-1 receptors (NK-1Rs) inhibited activity of N channels formed by Ca(V)2.2 and coexpressed with Ca(V)beta1b, Ca(V)beta3, or Ca(V)beta4 but enhanced activity of N channels containing Ca(V)beta2a. Exogenously applied AA produced the same pattern of modulation. Coexpression of Ca(V)beta2a, Ca(V)beta3, and Ca(V)beta4 recapitulated the modulatory response previously seen in SCG neurons, implying heterogeneous association of Ca(V)beta with Ca(V)2.2. Further experiments with mutated, chimeric Ca(V)beta subunits and free palmitic acid revealed that palmitoylation of Ca(V)beta2a is essential for loss of inhibition. The data presented here fit a model in which Ca(V)beta2a blocks inhibition, thus unmasking enhancement. Our discovery that the presence or absence of palmitoylated Ca(V)beta2a toggles M(1)R- or NK-1R-mediated modulation of N current between enhancement and inhibition identifies a novel role for palmitoylation. Moreover, these findings predict that at synapses, modulation of N-channel activity by M(1)Rs or NK-1Rs will fluctuate between enhancement and inhibition based on the presence of palmitoylated Ca(V)beta2a.

Ca2+ influx through both L- and N-type Ca2+ channels increases c-fos expression by electrical stimulation of sympathetic neurons.

During direct membrane depolarization, Ca2+ influx primarily through L-type Ca2+ (L-) channels initiates activity-dependent gene transcription. This is surprising given that in most neurons a minority of the total Ca2+ current arises from L-channel activity. However, many studies have stimulated Ca2+ influx with unphysiological stimuli such as chronic membrane depolarization using high K+ medium. Few studies have tested whether other Ca2+ channels stimulate gene transcription in adult neurons as a consequence of direct electrical stimulation. Therefore, we evaluated the role of L- and N-type Ca2+ (N-) channel activity in regulating mRNA levels of c-fos, an activity-dependent transcription factor, in adult rat superior cervical ganglion (SCG) neurons as the majority of Ca2+ channels are N-type, while only a minority are L-type. Changes in c-fos mRNA levels were measured using semi-quantitative and single-cell RT-PCR. Phosphorylation of CREB (pCREB) and changes in c-Fos levels were visualized in dissociated cells by immunocytochemistry. Increases in pCREB, c-fos mRNA and c-Fos protein with either K+ or electrical depolarization required Ca2+ influx. These results support previous findings that elevated c-fos levels result from pCREB stimulating c-fos transcription. Elevation of pCREB, c-fos and c-Fos with K+ depolarization depended on L-channel activity. By contrast, antagonizing either channel at 10-Hz stimulation minimized these increases despite unequal numbers of the two channel types. Transition to exclusive L-channel involvement occurred with increasing frequency of stimulation (from 10 to 20 to 50 Hz). Our results demonstrate that N- and L-channel participation in regulating c-fos expression is encoded in the pattern of electrical stimulation.

M1 muscarinic receptors inhibit L-type Ca2+ current and M-current by divergent signal transduction cascades.

Ion channels reside in a sea of phospholipids. During normal fluctuations in membrane potential and periods of modulation, lipids that directly associate with channel proteins influence gating by incompletely understood mechanisms. In one model, M(1)-muscarinic receptors (M(1)Rs) may inhibit both Ca(2+) (L- and N-) and K(+) (M-) currents by losing a putative interaction between channels and phosphatidylinositol-4,5-bisphosphate (PIP(2)). However, we found previously that M(1)R inhibition of N-current in superior cervical ganglion (SCG) neurons requires loss of PIP(2) and generation of a free fatty acid, probably arachidonic acid (AA) by phospholipase A(2) (PLA(2)). It is not known whether PLA(2) activity and AA also participate in L- and M-current modulation in SCG neurons. To test whether PLA(2) plays a similar role in M(1)R inhibition of L- and M-currents, we used several experimental approaches and found unanticipated divergent signaling. First, blocking resynthesis of PIP(2) minimized M-current recovery from inhibition, whereas L-current recovered normally. Second, L-current inhibition required group IVa PLA(2) [cytoplasmic PLA(2) (cPLA(2))], whereas M-current did not. Western blot and imaging studies confirmed acute activation of cPLA(2) by muscarinic stimulation. Third, in type IIa PLA(2) [secreted (sPLA(2))](-/-)/cPLA(2)(-/-) double-knock-out SCG neurons, muscarinic inhibition of L-current decreased. In contrast, M-current inhibition remained unaffected but recovery was impaired. Our results indicate that L-current is inhibited by a pathway previously shown to control M-current over-recovery after washout of muscarinic agonist. Our findings support a model of M(1)R-meditated channel modulation that broadens rather than restricts the roles of phospholipids and fatty acids in regulating ion channel activity.

[Effects of methionine synthase gene variation in parents on occurence of congenital heart disease in offspring].

OBJECTIVE: To investigate the relations of methionine synthase (MS) gene variation in parents with congenital heart disease (CHD) phenotype in offspring. METHODS: We selected 192 CHD patients (93 males and 99 females) and their biological parents (nuclear families) in Liaoning Province as case group, and 104 normal people (60 males and 44 females) and their parents without family history of birth defects as control group. For all subjects the MS gene A2756G locus polymorphism was examined by PCR-RFLP method. The data were analyzed to compare the difference of gene variation between the case parents and controls, and to assess the genetic disequilibrium in the CHD nuclear families. RESULTS: The MS genotype distribution and allele frequencies of the case mothers were not different significantly from those of the control group (P>0.05). The frequency of allele (+) in the case fathers (5.0%) was lower than that of the control (9.1%, P=0.060). The odds ratio (OR) was 0.53 (95% CI: 0.25-1.09). There was no difference in parents' genotype combination between the two groups. The analysis of allele transmission indicated that mutation allele (+) transmission disequilibrium existed in CHD nuclear families. The percentage of allele (+) transmitted from the parents was lower than (-) with being OR 0.26 (95% CI: 0.11-0.60). CONCLUSION: The study shows that the MS gene variation at A2756G locus in parents is associated with occurrence of CHD in offspring, and the mutation allele (+) in parents might be related with the decrease of CHD risk in offspring.

Polymorphism of methionine synthase gene in nuclear families of congenital heart disease.

OBJECTIVE: To investigate the relation of methionine synthase (MS) gene variation with congenital heart disease (CHD) phenotype. METHODS: One hundred and ninety three CHD patients (94 males and 99 females) and their biological parents (nuclear families) in Liaoning Province were selected as the case group, and another 104 normal persons (60 males and 44 females) and their parents without family history of birth defects as the control group. For all subjects the polymorphism of MS gene A2756G locus was examined by PCR-RFLP method. RESULTS: In offspring of the control group the frequencies of MS genotype (+/-) and allele (+) were 10.7% and 5.3%, without existence of homozygote. The MS genotype distribution and allele frequencies of CHD patients and their mothers were not significantly different from the control (P > 0.05). The frequency of allele (+) in case fathers (5.0%) was apparently lower than that in the control (9.1%, P = 0.060), and the odds ratio (OR) was 0.53 (95% CI: 0.25-1.09). There was no difference in parents' genotype combination between the two groups, and in genotype distribution among different types of CHD. Analysis of genetic transmission indicated that mutation allele (+) existed transmission disequilibrium in CHD nuclear families. The percentage of allele (+) transmitted from parents was lower than that allele (-) with OR 0.26 (95% CI: 0.11-0.60). CONCLUSION: MS gene variation in parents is associated with occurrence of CHD in offspring, and mutation allele (+) in parents may be related with the decrease of CHD risk in offspring.

[Relations of methionine synthase gene variation with congenital heart disease].

OBJECTIVE: Methionine synthase (MS) is the key enzyme in the homocysteine metabolism. To investigate the relations of MS gene variation with occurrence of congenital heart disease (CHD). METHODS: 186 CHD patients (0-31 years old) were selected as case group and 103 normal population as control. For all subjects the gene polymorphism at MS A2756G locus was analysed by PCR-RFLP method, and the serum folic acid/vitamin B12 levels were detected by radio-immunity assay. RESULTS: The heterozygotes (+/-) were detected in the subjects but without homozygotes (+/+). In control group the frequencies of (+/-) genotype and (+) allele were 10.7% and 5.3%, lower than Caucasian and Japanese population. In case group the frequencies of (+/-) genotype and (+) allele were 9.1% and 4.6%, without significantly different from control. The odds ratio of (+/-) genotype was 0.84 (0.35, 2.01). The genotype distributions in different types of CHD were also not apparently different with control. The serum vitamin B12 level was decreased in case group compared with control (336.66 pmol/L vs 465.72 pmol/L, P > 0.05), but the serum folic acid level no different. Also there were not significant difference for folic acid/vitamin B12 levels between different genotypes in case group. CONCLUSION: The results indicated that there was not apparent association between MS gene A2756G locus variation with CHD and serum folic acid/vitamin B12 levels. It need further investigations.

[Inhibition on the endocytosis of early visceral yolk sac induced by homocysteine in rats].

The effect of homocysteine (HCY) on the development and endocytosis of rat visceral yolk sac (VYS) was investigated. A preliminary method on early (GD9.5) rat yolk-sac placenta cultured in vitro was established by techniques using microdissection and transplantation. The results showed that HCY did damage VYS development and blood vessel formation, the damages were characterized by shrunk surface, small or defective blood island and significant reduction of diameter (P < 0.01). HCY also inhibited the process of VYS transport, specifically the endocytosis (as measured by [U-14C] sucrose uptake), the rate of endocytosis was significantly decreased when HCY was included in the medium (P < 0.01). These results suggested that the inhibition of sanguimotor development and the reduction of endocytosis induced by HCY may directly correlate with the teratologic and embryopathologic events.