Exploring the role and mechanism of Fuzi decoction in the treatment of osteoporosis by integrating network pharmacology and experimental verification.

BACKGROUND: Fuzi decoction (FZD), a traditional Chinese medicine formula, was used to treat musculoskeletal diseases by warming channels, strengthening yang and dispelling pathogenic cold and dampness. In clinical practice, FZD has been used to treat rheumatoid arthritis and osteoarthritis. It alleviated osteoarticular disorders through ameliorating the degradation of cartilage and improving meniscal damage in osteoarthritis, while its roles and mechanisms in the treatment of bone loss diseases remain unclear. This study aims to investigate the underlying mechanisms of FZD in treating osteoporosis using an integrative method of network pharmacology and experimental study. METHODS: In this study, network pharmacology was used to predict the core targets and potential pathways of the bioactive ingredients of FZD to attenuate osteoporosis. Molecular docking was performed to evaluate the interactions between core compounds and key targets. In addition, both cell and animal experiments were carried out to validate the role and potential mechanism in treating osteoporosis. RESULTS: In the present study, data revealed that kaempferol, beta-sitosterol, stigmasterol, fumarine, and (+)-catechin may be the primary bioactive ingredients of FZD in the treatment of osteoporosis, which were closely associated with the osteoporosis-related targets. And the KEGG results indicated that the NF-κB pathway was closely associated with the function of FZD in treating osteoporosis. In addition, in vivo demonstrated that FZD ameliorated osteoporosis. In vitro experiments showed that the pro-apoptotic factors indicators including CASP3 and BAX were decreased by FZD and the anti-apoptotic factor BCL2 was increased by FZD. In addition, FZD significantly suppressed the osteoclast differentiation in culture and the expression levels of osteoclast-related genes including TRAF6, CTSK, and MMP9. And the NF-κB pathway was confirmed, via in vitro experiment, to be involved in osteoclast differentiation. CONCLUSIONS: This study demonstrated that FZD played a pivotal role in suppressing the osteoclast differentiation via regulating the NF-κB pathway, indicating that FZD could be a promising antiosteoporosis drug and deserve further investigation.

Functional characterization, structural basis, and regio-selectivity control of a promiscuous flavonoid 7,4'-di-O-glycosyltransferase from Ziziphus jujuba var. spinosa.

A highly efficient and promiscuous 7,4'-di-O-glycosyltransferase ZjOGT3 was discovered from the medicinal plant Ziziphus jujuba var. spinosa. ZjOGT3 could sequentially catalyse 4'- and 7-O-glycosylation of flavones to produce 7,4'-di-O-glycosides with obvious regio-selectivity. For 7,4'-dihydroxyl flavanones and 3-O-glycosylated 7,4'-dihydroxyl flavones, ZjOGT3 selectively catalyses 7-O-glycosylation. The crystal structure of ZjOGT3 was solved. Structural analysis, DFT calculations, MD simulations, and site-directed mutagenesis reveal that the regio-selectivity is mainly controlled by the enzyme microenvironment for 7,4'-dihydroxyl flavones and 3-O-glycosylated 7,4'-dihydroxyl flavones. For 7,4'-dihydroxyl flavanones, the selectivity is mainly controlled by intrinsic reactivity. ZjOGT3 is the first plant flavonoid 7,4'-di-O-glycosyltransferase with a crystal structure. This work could help understand the catalytic mechanisms of multi-site glycosyltransferases and provides an efficient approach to synthesise O-glycosides with medicinal potential.

Effects of L-carnitine Administration on Sperm and Sex Hormone Levels in a Male Wistar Rat Reproductive System Injury Model in a High-Altitude Hypobaric Hypoxic Environment.

The plateau environment impacts male reproductive function, causing decreased sperm quality and testosterone levels. L-carnitine can improve the semen microenvironment. However, the role of L-carnitine in a high-altitude environment remains unclear. In our study, we investigated the effects of L-carnitine administration in a male Wistar rat reproductive system injury model in the context of a simulated high-altitude environment. Rats were randomly divided into a normal control group (group A1, A2-low dose and A3-high dose) and high-altitude model groups (group B, C-low dose and D-high dose) with 20 rats in each group. With the exception of the normal control group exposed to normoxic conditions, the other groups were maintained in a hypobaric oxygen chamber that simulated an altitude of 6000 m for 28 days. In the experimental period, the low-dose groups (A2 and C) were administered 50 mg/kg L-carnitine via intraperitoneal injection once a day, and the high-dose groups (A3 and D) were given 100 mg/kg. After the feeding period, blood samples were collected to assess blood gas, serum hormone levels and oxidative stress. Sperm from the epididymis were collected to analyse various sperm parameters. After obtaining the testicular tissue, the morphological and pathological changes were observed under a light microscope and transmission electron microscopy (TEM). The impact of the simulated high-altitude environment on the rat testis tissue is obvious. Specifically, a decreased testicular organ index and altered indices of arterial blood gas and serum sex hormone levels caused testicular tissue morphological damage, reduced sperm quality, increased sperm deformity rate and altered malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) concentrations. The results demonstrate that L-carnitine can be administered as a preventive intervention to reduce the reproductive damage caused by high-altitude hypobaric and hypoxic environments and improve semen quality in a rat model.

Functional Characterization and Protein Engineering of a Triterpene 3-/6-/2'-O-Glycosyltransferase Reveal a Conserved Residue Critical for the Regiospecificity.

Engineering the function of triterpene glucosyltransferases (GTs) is challenging due to the large size of the sugar acceptors. In this work, we identified a multifunctional glycosyltransferase AmGT8 catalyzing triterpene 3-/6-/2'-O-glycosylation from the medicinal plant Astragalus membranaceus. To engineer its regiospecificity, a small mutant library was built based on semi-rational design. Variants A394F, A394D, and T131V were found to catalyze specific 6-O, 3-O, and 2'-O glycosylation, respectively. The origin of regioselectivity of AmGT8 and its A394F variant was studied by molecular dynamics and hydrogen deuterium exchange mass spectrometry. Residue 394 is highly conserved as A/G and is critical for the regiospecificity of the C- and O-GTs TcCGT1 and GuGT10/14. Finally, astragalosides III and IV were synthesized by mutants A394F, T131V and P192E. This work reports biocatalysts for saponin synthesis and gives new insights into protein engineering of regioselectivity in plant GTs.

Physical Work and Exercise Reduce the Risk of Cognitive Impairment in Older Adults: A Population-based Longitudinal Study.

BACKGROUND: The independent effect of physical work on the risk of cognitive impairment in older Chinese older adults living in rural areas remains to be elucidated. OBJECTIVE: We aimed to determine whether physical work and physical exercise can reduce the risk of cognitive impairment. METHODS: We collected data from 7,000 permanent residents without cognitive impairment (age ≥60 years) over a follow-up period of 2 years. We used the Chinese version of the Mini-Mental State Examination (MMSE) to assess cognitive function. We performed multivariate Cox regression analyses to calculate adjusted Hazard Ratios (HRs) and 95% Confidence Intervals (%95 CIs) as measures of the association between physical work/exercise and cognitive impairment while controlling for potential confounders. RESULTS: Over a median follow-up period of 1.93 years, 1,224 (17.5%) of 7,000 participants developed cognitive impairment, with a total incidence of 97.69 per 1,000 person-years. After adjustment for potential confounders, participating in physical work (HR: 0.51; 95% CI: 0.43-0.60) or physical exercise (HR: 0.53; 95% CI: 0.44-0.65) was associated with a reduced risk of cognitive impairment. Stratified analyses suggested additive and multiplicative interactions between physical work and exercise. Agricultural work (HR: 0.46; 95% CI: 0.38-0.55), walking/tai chi (HR: 0.54; 95% CI: 0.44-0.67), and brisk walking/yangko (HR: 0.57; 95% CI: 0.33-0.97) exerted significant protective effects against cognitive impairment. CONCLUSION: Both physical work and exercise can reduce the risk of cognitive impairment in older adults. Reasonable types and appropriate intensities of physical activity are recommended to prevent or delay the progression of cognitive impairment.

Functional Characterization and Structural Basis of an Efficient Di-C-glycosyltransferase from Glycyrrhiza glabra.

A highly efficient di-C-glycosyltransferase GgCGT was discovered from the medicinal plant Glycyrrhiza glabra. GgCGT catalyzes a two-step di-C-glycosylation of flopropione-containing substrates with conversion rates of >98%. To elucidate the catalytic mechanisms of GgCGT, we solved its crystal structures in complex with UDP-Glc, UDP-Gal, UDP/phloretin, and UDP/nothofagin, respectively. Structural analysis revealed that the sugar donor selectivity was controlled by the hydrogen-bond interactions of sugar hydroxyl groups with D390 and other key residues. The di-C-glycosylation capability of GgCGT was attributed to a spacious substrate-binding tunnel, and the G389K mutation could switch di- to mono-C-glycosylation. GgCGT is the first di-C-glycosyltransferase with a crystal structure, and the first C-glycosyltransferase with a complex structure containing a sugar acceptor. This work could benefit the development of efficient biocatalysts to synthesize C-glycosides with medicinal potential.

Effectiveness and safety of Shenmai injection in treatment of shock: A Meta-analysis.

OBJECTIVE: To evaluate the effectiveness and safety of Shenmai injection for shock. METHODS: Randomized controlled trials (RCTs) that evaluated the therapeutic effect of Shenmai injection on shock (including septic shock, cardiogenic shock, hypovolemic shock, neurogenic shock and anaphylactic shock) were included in this analysis. The major electronic databases were searched until May 2015. The methodological quality of the trials was assessed according to the Cochrane Handbook. Review Manager 5.3 and Stata 12.0 software were applied for data analysis. RESULTS: Thirty RCTs involving 2038 participants were included. The methodological quality of the trials was generally passable. The combined use of Shenmai injection and conventional medicine was significantly more effective at managing shock compared to conventional medicine alone in the outcomes of total effective rate [risk ratio (RR) 1.25, 95% confidence interval (CI) 1.18 to 1.31] and mortality rate [risk difference (RD) -0.10, 95% CI -0.17 to -0.02]. Likewise, improvements were observed in other metrics. Three trials reported adverse events, but no trial reported serious adverse effects. CONCLUSION: Our results indicated the potential effectiveness of Shenmai injection combined with conventional medicine treatment for shock. However, further rigorously designed trials are needed to collect and weigh up all the evidence for the use of Shenmai injection.

Tea consumption is inversely associated with depressive symptoms in the elderly: A cross-sectional study in eastern China.

BACKGROUND: Epidemiological studies suggest that higher tea consumption was associated with lower risk of depressive symptoms, but this has not been found consistently. Moreover, the effect of different types of tea on depressive symptoms needs to be further explored. This study aimed to examine the association between tea consumption and depressive symptoms in Chinese elderly. METHODS: We analyzed the baseline data from Zhejiang Major Public Health Surveillance Program including 9371 participants. Depressive symptoms was assessed through the application of Patient Health Questionnaire-9 scale (PHQ-9). Logistic regression models, controlled for an extensive range of potential confounders, were generated to evaluate the association between tea consumption and risk of depressive symptoms. RESULTS: The black tea drinkers had a significantly decreased risk of depressive symptoms (p<0.01), whereas no association was found in green tea drinkers. Compared with non-drinkers, the adjusted ORs (95% CIs) were 0.48 (0.23, 0.99) and 0.35 (0.17, 0.72) for participants consuming <3 cups and ≥3 cups of black tea per day, respectively (P for trend: <0.01). A linear association between concentration of black tea and depressive symptoms was also confirmed in our study. LIMITATIONS: Cross-sectional data could not make a causation conclusion, and the observed association in our study could not be ascribed to any specific component in tea. CONCLUSIONS: Our results indicated that higher black tea consumption was associated with a lower prevalence of depressive symptoms in the elderly.

Tea Consumption and Cognitive Impairment: A Cross-Sectional Study among Chinese Elderly.

BACKGROUND: Laboratorial and epidemiological researches suggested that tea exhibited potential neuroprotective effect which may prevent cognitive impairment, but there were few data among the elderly aged 60 years and above in China. OBJECTIVE: The objective was to explore the relationship between characteristics of tea consumption and cognitive impairment. DESIGN: We analyzed the baseline data from Zhejiang Major Public Health Surveillance Program (ZPHS) which was conducted in 2014. Totally 9,375 residents aged 60 years and above were recruited in this study. Face-to-face interview based on a self-developed questionnaire was performed for each participant. Detailed tea consumption habits were included in the questionnaire. Cognitive impairment screening was performed by using Mini-Mental State Examination (MMSE). Education-specific cut-off points for Chinese were applied to determine the status of cognitive impairment. Logistic regression analysis was used to calculate odds ratios (ORs) of cognitive impairment associated with tea consumption. RESULTS: The means (SD) of MMSE scores for the subjects who did not consume tea and consumed <2 cups/d, 2-4 cups/d, ≥4 cups/d were 23.3 (SD = 5.61), 23.8 (SD = 5.60), 24.5 (SD = 5.63) and 25.0 (SD = 5.08), respectively. An inverse correlation was found between tea consumption (of all types) and prevalence of cognitive impairment. Volume of tea consumption was significantly associated with cognitive impairment: compared with non-consumption participants, those who consumed < 2 cups/d, 2-4 cups/d, and ≥4 cups/d were observed ORs of 0.77 (95% CI: 0.56, 1.07), 0.62 (95% CI: 0.47, 0.81), and 0.49 (95% CI: 0.36, 0.66), respectively. Compared with non-consumption, black tea presented a positive correlation with cognitive function after controlling for potential confounders (OR = 0.52, 95% CI: 0.28, 0.95), while green tea showed no significant difference (OR = 1.04, 95% CI: 0.72, 1.51). Participants who consumed weak tea, moderate tea or strong tea more often were observed a better cognitive status when compared with those who did not have tea, with an OR of 0.51 (95% CI: 0.28, 0.92), 0.32 (95% CI: 0.19, 0.56) and 0.42 (95% CI: 0.22, 0.78) after adjusting for the potential confounders. But there was no statistically significant difference between any two of these ORs. CONCLUSION: Black tea consumption was association with better cognitive performance among the elderly aged 60 years and above in China, while green tea presented no correlation. The positive association of cognitive status with tea consumption was not limited to particular type of concentration.

An outbreak of Mycobacterium tuberculosis infection associated with acupuncture in a private clinic of Zhejiang Province, China, 2012.

BACKGROUND: Acupuncture carries the potential risk of transmission of pathogenic microorganisms from the environment to the patient, and from one patient to another. An outbreak of tuberculosis at a private clinic in eastern China was investigated to identify the source of infection, mode of transmission, and risk factors for infection. METHODS: A probable case was one who had the onset of unexplained pain, swelling, or abscess in the area of invasive treatment between January 1, 2011 and February 23, 2012. A confirmed case was a probable case with positive laboratory test results for Mycobacterium tuberculosis. Patient history and the frequency of invasive treatment were compared between 56 probable and confirmed cases and 98 controls in a case-control study. RESULTS: Fifty-six of 2561 patients (2.2%) who had visited the clinic developed tuberculosis. The odds ratio (OR) of M. tuberculosis infection increased with the frequency of clinic visits (Chi-square for trend=28.943, p=0.000). Multivariate analysis showed that the frequency of acupuncture (Chi-square=24.258, adjusted p-value=0.000) and sharing acupuncture needles (Chi-square=8.936, adjusted p-value=0.003) were risk factors for M. tuberculosis infection. Thirty-two pus sample and nine sputum sample cultures were M. tuberculosis-positive. CONCLUSIONS: This outbreak was caused by acupuncture and was transmitted through sharing acupuncture needles contaminated with M. tuberculosis.

[Meta-analysis on efficacy and safety of Shenmai injection in treatment of coronary heart disease].

OBJECTIVE: To evaluate the efficacy and safety of Shenmai injection in the treatment of coronary heart disease (CHD). METHOD: CNKI (1979 to December 2011), VIP (1989 to December 2011), and Wanfang Data (1987 to December 2011) were retrieved for literatures concerning randomized controlled trials of Shenmai injection in the treatment of coronary heart disease. Their quality was assessed by the Jadad scale. Review Manager 5.0 and Stata 12 software were adopted for data analysis. RESULT: Altogether 18 randomized clinical trials were included in the Meta-analysis. According to the findings, compared with routine therapy alone, the combination of Shenmai injection and routine therapy increased the overall effective rate by clinical symptoms by 22% [RD = 0.22, 95% CI (0.17, 0.26), P<0.000 01], with the relative risk ratio of [RR = 1.31, 95% CI (1.20, 1.43), P<0.000 1] and the efficacy by electrocardiogram by 25% [RD = 0.25, 95% CI (0.19, 0.31), P<0.000 01], with the relative risk ratio of [RR = 1.46, 95% CI (1.32, 1.62), P<0.000 1]. It also improved New York heart association classification (NYHA) by 33% [RD = 0.33, 95% CI (0.16, 0.49), P<0.000 01], with the relative risk ratio of [RR = 1.79, 95% CI (1.28, 2.51), P = 0.000 6] and left ventricular ejection fraction [WMD 7.55, 95% CI (2.28, 12.83)]. Additionally, studies reported that Shenmai injection could increase left ventricular stroke volume, decrease left ventricular end systolic diameter, improved hemodynamic indexes, reduced NT-pro-BNP, DMA, cytokines, TNF-alpha, IL-6 and serum enzyme, mitigated infarction-related revascularization IRA indexes and shortened the symptom improvement time and electrocardiogram recovery time. Three studies had reported adverse events, mainly dizziness, palpitation, mild abdominal distension, nausea, metal excitement and flushed face, with no serious adverse effect. CONCLUSION: Clinical evidences prove that Shenmai injection has certain effect in treating CHD and remains to be improved in the overall effective rate. The essay, despite of some shortcomings, provides certain reference for clinical application. It is suggested holding more multicenter randomized controlled trials with rational design, strict implementation, large sample size and sufficient following-up time to further verify its efficacy and safety.