Direct Observation of the Mutual Coupling Effect in the Protein-Water-Glycerol Mixture by Combining Neutron Scattering and Selective Deuteration.

Numerous studies have discussed the impact of cosolvents on the structure, dynamics, and stability of proteins in aqueous solutions. However, the dynamics of cosolvents in the protein-water-cosolvent ternary system is largely unexplored in experiments due to technical difficulty. Consequently, a comprehensive understanding of the interplay among proteins, water, and cosolvents is still lacking. Here, we employed selective deuteration and neutron scattering techniques to characterize the individual motions of each component in the protein/water/glycerol (GLY) mixture across various temperatures. The consistent dynamic onset temperatures and the correlation between the MSD of the protein and the viscosity of solvents revealed the mutual coupling effects among the three components. Furthermore, our experimental and simulation results showed that the hydrogen bond relaxation energy barrier in the ternary system is ∼43 kJ/mol, whereas in the protein-water binary system it is merely ∼35 kJ/mol. Therefore, we suggest that GLY can enhance hydrogen bond interactions in the ternary system through the mutual coupling effect, thereby serving as one of the protective mechanisms of protein preservation by GLY.

High-Dose Clopidogrel versus Ticagrelor in CYP2C19 intermediate or poor metabolizers after percutaneous coronary intervention: A Meta-Analysis of Randomized Trials.

WHAT IS KNOWN AND OBJECTIVE: For patients after percutaneous coronary interventions (PCI), clopidogrel combined with aspirin is a conventional dual antiplatelet therapy (DAPT) method. Because the genetic polymorphism of CYP2C19 gene leads to clopidogrel resistance, guidelines for antiplatelet recommendations in CYP2C19 of ultrarapid metabolizers (UM), extended metabolizers (EM) and poor metabolizers (PM) are clear. However, there is no clear recommendation as to whether ticagrelor or double dose clopidogrel is the best antiplatelet regimen for CYP2C19 of intermediate metabolizers (IM). To evaluate the efficacy and safety of ticagrelor (combined with aspirin) and high-dose clopidogrel (combined with aspirin) in patients after PCI with CYP2C19 loss-of-function (LOF) alleles. METHODS: We searched the following databases to select RCTs of comparing ticagrelor with high-dose clopidogrel in patients after PCI with CYP2C19 LOF alleles: CNKI, Wanfang Data, PubMed, Clinical trials, Cochrane, Web of Science and Embase. Major adverse cardiovascular events (MACEs), platelet function and TIMI bleeding event were defined as the outcomes. revman 5.3 software was used to perform meta-analysis. RESULTS AND DISCUSSION: A total of 14 RCTs with 2351 patients were enrolled. Meta-analysis showed that compared with high-dose clopidogrel, ticagrelor had reduced incidence of MACEs (OR = 0.32, 95% Cl: 0.23-0.44, p < 0.00001), stent thrombosis (OR: 0.24, 95%CI: 0.13-0.44, p < 0.00001), myocardial infarction OR: 0.42, 95%CI: 0.22-0.80, p = 0.008), revascularization (OR: 0.29, 95%CI: 0.10-0.82, p = 0.02) and unstable angina (OR: 0.47, 95%CI: 0.29-0.77, p = 0.003) in patients after PCI with CYP2C19 LOF alleles. A subgroup analysis showed that ticagrelor reduced the risk of MACEs compared with high-dose clopidogrel regardless of the type of metabolizer. Compared with high-dose clopidogrel, ticagrelor significantly reduced the risk of MACE with longer follow-up period (more than 3 months) without increasing the risk of bleeding (OR: 0.89, 95%CI: 0.53-1.49, p = 0.30), while elevated dyspnoea (OR: 5.62, 95%CI: 3.07-10.28, p < 0.00001). WHAT IS NEW AND CONCLUSIONS: For patients carrying CYP2C19 LOF alleles after PCI, ticagrelor may be better than high-dose clopidogrel in reducing the risk of MACEs, while dyspnoea incidents should be alerted.

Neuroprotective effects of Shende'an tablet in the Parkinson's disease model.

BACKGROUND: Shende'an tablet (SDA) is a newly capsuled Chinese herbal formula derived from the Chinese traditional medicine Zhengan Xifeng Decoction which is approved for the treatment of neurasthenia and insomnia in China. This study aimed to investigate the neuroprotective effects of SDA against Parkinson's disease (PD) in vitro and in vivo. METHODS: In the present work, the neuroprotective effects and mechanism of SDA were evaluated in the cellular PD model. Male C57BL/6J mice were subject to a partial MPTP lesion alongside treatment with SDA. Behavioural test and tyrosine-hydroxylase immunohistochemistry were used to evaluate nigrostriatal tract integrity. HPLC analysis and Western blotting were used to assess the effect of SDA on dopamine metabolism and the expression of HO-1, PGC-1α and Nrf2, respectively. RESULTS: Our results demonstrated that SDA had neuroprotective effect in dopaminergic PC12 cells with 6-OHDA lesion. It had also displayed efficient dopaminergic neuronal protection and motor behavior alleviation properties in MPTP-induced PD mice. In the PC12 cells and MPTP-induced Parkinson's disease animal models, SDA was highly efficacious in α-synuclein clearance associated with the activation of PGC-1α/Nrf2 signal pathway. CONCLUSIONS: SDA demonstrated potential as a future therapeutic modality in PD through protecting dopamine neurons and alleviating the motor symptoms, mediated by the activation of PGC-1α/Nrf2 signal pathway.

Detection of pelvic lymph node micrometastasis by real-time reverse transcriptase polymerase chain reaction in prostate cancer patients after hormonal therapy.

OBJECTIVE: To determine the feasibility of prostatic-specific antigen (PSA) mRNA and prostatic-specific membrane antigen (PSMA) mRNA measurement in detection of pelvic lymph node (PLN) micrometastasis for prostate cancer (PCa) after hormonal therapy (HT). METHODS: Fifty-four patients diagnosed as high risk localized PCa were given HT for 3 months before radical prostatectomy. Under bipedal lymphangiography, a needle was punctured into involved lymph nodes (LN) and aspirated lymphatic fluid was obtained preoperatively. The expression of PSA mRNA and PSMA mRNA in aspirated fluid was assessed by a fully quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR) and also in LN specimens from pelvic lymphadenectomy during prostatectomy. RESULTS: Median follow-up was 36 months (range 18-58 months). Without histological evidence of PLN metastasis, twelve patients showed positive PSA and/or PSMA mRNA expressions and regarded as having micrometastases to PLNs. Biochemical recurrence (BCR) rate and interval between prostatectomy and BCR in patients with micrometastases (group B) were not significantly different to histologically proven PLN metastatic patients (group A) (58.3 vs. 83.3 %, P = 0.26; 10.9 vs. 9.2 months, P = 0.29, respectively), but significantly different to those with no PLN involvement (group C) (58.3 vs. 11.1 %, P = 0.002; 10.9 vs. 21.3 months, P < 0.001, respectively). Kaplan-Meier analysis showed both groups A and B had significantly lower non-BCR rate than group C (P < 0.001, P < 0.001, respectively). CONCLUSIONS: For PCa patients receiving HT, measurement of PSA mRNA and PSMA mRNA in aspirated PLN fluid by real-time RT-PCR could effectively detect PLN micrometastases without surgical intervention.

[Study on micrometastases in pelvic lymph nodes detected by real-time reverse transcriptase polymerase chain reaction with clinically localized prostate cancer before radical prostatectomy].

OBJECTIVE: To clarify the significance of micrometastases in pelvic lymph nodes in patients with neoadjuvant hormonal therapy (NHT) before radical prostatectomy (RP). METHODS: Twenty-one patients with clinically localized prostate cancer who received NHT between August 2007 and March 2010 were observed. The patients were clarified into four groups: pathological examination was positive (group A), real-time PCR examination targeting prostate specific antigen (PSA) mRNA and prostate specific membrane antigen (PSMA) mRNA were positive (group B), pathological examination and real-time PCR examination targeting PSA mRNA and PSMA mRNA were both negative (group C), and the control group (group D). After a standard bipedal lymphangiography the films were reviewed carefully by an experienced radiologist. If positive lymph nodes were seen or suspected, a thin-walled 22 gauge needle were directed transabdominally under fluoroscopic control into the area of question and an aspirate was obtained. The expression of PSA and PSMA in aspirate were assessed by a fully quantitative real-time PCR. The specimens were regarded in which either PSA mRNA or PSMA mRNA were positive as showing the "presence of micrometastasis". Lymph node specimens were also stained immunohistochemically with an antibody PSA after RP. RESULTS: Pathological examination detected lymph node metastases from 3 cases, and real-time PCR further identified lymph node micrometastases from 14 cases with no pathological evidence of nodal involvement. The expression level of PSA mRNA and PSMA mRNA were statistically significant in patients with histological confirmed lymph node metastases and micrometastases detected by real-time PCR despite the lack of histological evidence, and the expression level of PSA mRNA and PSMA mRNA in aspirate were higher than the lymph node between the group A and group B. CONCLUSIONS: Although residual foci of atrophic prostate cancer cells in resected lymph nodes after NHT can be difficult to diagnose by pathological examination, the present results show the usefulness of quantitative real-time PCR targeting PSA and PSMA mRNA for detected micrometastatic tumour foci in pelvic lymph nodes from fine needle aspiration biopsy of lymph nodes before RP.

[Clinical outcomes of sexuality preserving cystectomy and Roux-y sigmoid neobladder].

OBJECTIVE: To assess the penile erectile function, urinary continence and voiding, and cancer recurrence in 18 bladder cancer patients after sexuality preserving cystectomy and Roux-y sigmoid neobladder reconstruction. METHODS: Eighteen male patients with bladder cancer underwent sexuality preserving cystectomy and Roux-y sigmoid neobladder reconstruction, and were followed up for cancer recurrence and such clinical outcomes as erectile function and urinary continence and voiding. RESULTS: The patients were followed up for an average of 41 months, of whom, all achieved day- and night-time urinary continence, but 2 with positive lymph nodes died of extensive metastasis at 10 and 15 months, respectively, after surgery. Among the total number, potency was maintained in 11 patients (61.1%), impaired in 2 and lost in 5, and the post-operative IIEF-5 score was (10.83 +/- 8.25) as compared with (13.72 +/- 6.39) before the operation, with a statistically significant difference (P < 0.05). CONCLUSION: Erectile function and urinary continence are fairly good in bladder cancer patients after sexuality preserving cystectomy and Roux-y sigmoid neobladder reconstruction, and the oncological results are fairly acceptable, but still need to be confirmed by longer follow-ups and larger trials.