Cerebrovascular diseases have the characteristics of high morbidity, high disability, high mortality and high recurrence rate, which seriously harm human health and increase the national health economic burden. 3-n-Butylphthalide (NBP) is a new drug commonly used in clinical treatment of cerebrovascular diseases, and it is also one of the main active components in traditional Chinese medicine such as Angelica sinensis and Chuanxiong. In this review, the pharmacological effects of NBP were systematically summarized. Studies have shown that NBP has pharmacological effects such as antiplatelet aggregation, anti-thrombosis, inhibiting neuronal apoptosis, anti-oxidation, anti-cerebral ischemia, anti-brain injury, and anti-vascular dementia. In clinical practice, it is often combined with edaravone, alteplase, fasudil, Xingnaojing injection, and compound Danshen injection to treat cerebral vascular diseases such as stroke, vascular dementia, and cerebral vasospasm, and plays a good synergistic effect. This summary could provide support for the rational clinical application of NBP, and also provide basis for the in-depth study of the interaction of its drug combination.

OBJECTIVE: To investigate the effects of matrine combined with LY294002 on proliferation, apoptosis and cell cycle of human myeloid leukemia K562 cells and explore the underlying mechanism. METHODS: The effects of different concentrations of matrine alone and in combination with LY294002 on the proliferation of K562 cells were examined with CCK-8 assay. The changes in morphology of K562 cells were observed following treatment for 48 h with 0.4 g/L matrine and 10 μmol/L Y294002, either alone or in combination, and cell apoptosis was detected using flow cytometry with annexin V-FITC/PI double labeling; the changes in cell cycle was detected with PI labeling. Western blotting was performed to examine the effect of matrine combined with LY294002 on expressions of p-mTOR, p-PI3K, Akt, p-Akt, cyclinD1, Bcl-2 and caspase-9 in the cells. RESULTS: Treatment with different concentrations of matrine, both alone and in combination with LY294002, inhibited the proliferation of K562 cells in a time- and concentration-dependent manner. Compared with matrine treatment alone, the combined treatment caused more obvious morphological changes of the cells, significantly increased cell apoptosis (P < 0.01), and induced cell cycle arrest in G₀/G₁ (P < 0.01). Western blotting showed that the protein expression levels of p-mTOR, cyclinD1, p-PI3K, p-Akt and Bcl-2 in K562 cells increased while the expression level of caspase-9 decreased significantly after the combined treatment (P < 0.01). CONCLUSION Matrine combined with LY294002 produces a synergistic inhibitory effect on K562 cells possibly by down-regulating the p-Akt expression in PI3K/Akt signaling pathway, reducing the expressions of p-mTOR, cyclinD1 and Bcl-2, and increasing the expression of caspase-9.
Humans ; K562 Cells ; Caspase 9 ; Matrines ; Phosphatidylinositol 3-Kinases ; Cell Cycle ; Cell Division ; Leukemia, Myeloid ; Apoptosis ; Proto-Oncogene Proteins c-bcl-2

CRISPR-Cas9 gene editing technology has been widely used in Saccharomyces cerevisiae.However, the effects of Cas9, as an exogenous protein, on the growth and production of natural products in S.cerevisiae are still unclear.In this study, Cas9 gene was expressed in S.cerevisiae by integration into the genome and construction into vectors, and two natural products, carotenoid and miltiradiene, were selected as the target products to study the effects of Cas9 expression on yeast growth and production capacity.The results showed that whether Cas9 was integrated into the genome or expressed by vectors, Cas9 inhibited the growth of S.cerevisiae, which was more obvious in the form of genome integration.When Cas9 was integrated into the genome, it had no effect on the production of carotenoid and miltiradiene by S.cerevisiae, but when Cas9 was expressed by vectors, the ability of S.cerevisiae to produce carotenoids and miltiradiene was significantly reduced.Therefore, in order to further efficiently knock out Cas9 after gene editing and minimize the adverse impact of Ura3 and Trp1 vectors, this study systematically explored the removal efficiency of the two vectors, and a plasmid capable of efficient gene editing was constructed, which optimized the application of CRISPR-Cas9 gene editing system in S.cerevisiae, and provided reference for the application of gene editing technology based on Cas9.
Biological Products ; CRISPR-Cas Systems ; Carotenoids/metabolism* ; Gene Editing/methods* ; Saccharomyces cerevisiae/metabolism*

OBJECTIVE: To analyze the clinical characteristics of multiple myeloma(MM) patients with early relapse. METHODS: A total of 50 MM patients with early relapse (≤12 months) and 50 matched controls with late relapse (>12 months) were selected. The time from diagnosis to relapse and related clinical data of the 100 patients were retrospectively analyzed, and the factors associated with early relapse were identified. Kaplan-Meier curve was used to analyze the overall survival (OS) time of the whole cohort. Area under the curve (AUC) was used to evaluate the effect of circulating plasma cells on early recurrence of the patients. RESULTS: The results showed that high-risk cytogenetics (FISH) (P=0.005), and ISS stage III (P=0.008) were associated with early recurrence of the patients. For patients with early relapse, high-risk FISH showed poor survival. Compared with the patients with late relapse, most of the chromosome karyotype of patients with early relapse showed quantitative and structural abnormalities. The expression of circulating plasma cells was significantly increased in patients with early recurrence group (P=0.0318). The response to initial treatment was poor in the early recurrence group (P=0.001), and the prognosis was significantly worse than those in the late recurrence group (median OS: 38 vs 81 months, P=0.002). CONCLUSION Early relapse is a marker poor prognostic in MM patients, and such patients should be focused on the improving their prognosis.
Cytogenetics ; Humans ; Multiple Myeloma/drug therapy* ; Neoplasm Recurrence, Local ; Prognosis ; Retrospective Studies

In order to clarify the influence of acute hypobaric hypoxia on the bile acids of the rat small intestine, we used ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) to identify bile acids in the contents of the small intestine from untreated and acute hypobaric hypoxia-treated rats. Thirty-nine bile acids were detected; PCA and OPLS-DA analysis revealed marked differences in the composition of bile acids between the untreated and the acute hypobaric hypoxia groups. Bile acids were screened with VIP > 1, |log₂FC| ≥ 1, P < 0.05, and a total of 7 bile acids with significant differences in content between the two groups were obtained, including 5 conjugated bile acids, 2 unconjugated bile acids; in addition, the content of conjugated bile acids has risen in the treated group. This study demonstrated the influence of high-altitude hypoxic environment on bile acid composition and metabolism in rats. All the animal experiments in this study were approved by the 940th Hospital Ethics Committee (approval No: 2020KYLL012).

BACKGROUND: Innovative coronavirus disease 2019 (COVID-19) vaccines, with elevated global manufacturing capacity, enhanced safety and efficacy, simplified dosing regimens, and distribution that is less cold chain-dependent, are still global imperatives for tackling the ongoing pandemic. A previous phase I trial indicated that the recombinant COVID-19 vaccine (V-01), which contains a fusion protein (IFN-PADRE-RBD-Fc dimer) as its antigen, is safe and well tolerated, capable of inducing rapid and robust immune responses, and warranted further testing in additional clinical trials. Herein, we aimed to assess the immunogenicity and safety of V-01, providing rationales of appropriate dose regimen for further efficacy study. METHODS: A randomized, double-blind, placebo-controlled phase II clinical trial was initiated at the Gaozhou Municipal Centre for Disease Control and Prevention (Guangdong, China) in March 2021. Both younger (n = 440; 18-59 years of age) and older (n = 440; ≥60 years of age) adult participants in this trial were sequentially recruited into two distinct groups: two-dose regimen group in which participants were randomized either to follow a 10 or 25 μg of V-01 or placebo given intramuscularly 21 days apart (allocation ratio, 3:3:1, n = 120, 120, 40 for each regimen, respectively), or one-dose regimen groups in which participants were randomized either to receive a single injection of 50 μg of V-01 or placebo (allocation ratio, 3:1, n = 120, 40, respectively). The primary immunogenicity endpoints were the geometric mean titers of neutralizing antibodies against live severe acute respiratory syndrome coronavirus 2, and specific binding antibodies to the receptor binding domain (RBD). The primary safety endpoint evaluation was the frequencies and percentages of overall adverse events (AEs) within 30 days after full immunization. RESULTS: V-01 provoked substantial immune responses in the two-dose group, achieving encouragingly high titers of neutralizing antibody and anti-RBD immunoglobulin, which peaked at day 35 (161.9 [95% confidence interval [CI]: 133.3-196.7] and 149.3 [95%CI: 123.9-179.9] in 10 and 25 μg V-01 group of younger adults, respectively; 111.6 [95%CI: 89.6-139.1] and 111.1 [95%CI: 89.2-138.4] in 10 and 25 μg V-01 group of older adults, respectively), and remained high at day 49 after a day-21 second dose; these levels significantly exceed those in convalescent serum from symptomatic COVID-19 patients (53.6, 95%CI: 31.3-91.7). Our preliminary data show that V-01 is safe and well tolerated, with reactogenicity predominantly being absent or mild in severity and only one vaccine-related grade 3 or worse AE being observed within 30 days. The older adult participants demonstrated a more favorable safety profile compared with those in the younger adult group: with AEs percentages of 19.2%, 25.8%, 17.5% in older adults vs. 34.2%, 23.3%, 26.7% in younger adults at the 10, 25 μg V-01 two-dose group, and 50 μg V-01 one-dose group, respectively. CONCLUSIONS: The vaccine candidate V-01 appears to be safe and immunogenic. The preliminary findings support the advancement of the two-dose, 10 μg V-01 regimen to a phase III trial for a large-scale population-based evaluation of safety and efficacy. TRIAL REGISTRATION http://www.chictr.org.cn/index.aspx (No. ChiCTR2100045107, http://www.chictr.org.cn/showproj.aspx?proj=124702).
Aged ; Antibodies, Viral ; COVID-19/therapy* ; COVID-19 Vaccines ; Double-Blind Method ; Humans ; Immunization, Passive ; Recombinant Fusion Proteins ; SARS-CoV-2

TANK-binding kinase 1 (TBK1), a core kinase of antiviral pathways, activates the production of interferons (IFNs). It has been reported that deacetylation activates TBK1; however, the precise mechanism still remains to be uncovered. We show here that during the early stage of viral infection, the acetylation of TBK1 was increased, and the acetylation of TBK1 at Lys241 enhanced the recruitment of IRF3 to TBK1. HDAC3 directly deacetylated TBK1 at Lys241 and Lys692, which resulted in the activation of TBK1. Deacetylation at Lys241 and Lys692 was critical for the kinase activity and dimerization of TBK1 respectively. Using knockout cell lines and transgenic mice, we confirmed that a HDAC3 null mutant exhibited enhanced susceptibility to viral challenge via impaired production of type I IFNs. Furthermore, activated TBK1 phosphorylated HDAC3, which promoted the deacetylation activity of HDAC3 and formed a feedback loop. In this study, we illustrated the roles the acetylated and deacetylated forms of TBK1 play in antiviral innate responses and clarified the post-translational modulations involved in the interaction between TBK1 and HDAC3.

Objective: To observe the clinical efficacy of DZWJY-1 type electronic moxibustion apparatus and traditional moxibustion in treating knee osteoarthritis (KOA). Methods: A total of 76 eligible patients were randomized into an electronic moxibustion apparatus group and a traditional moxibustion group, with 38 cases in each group. The electronic moxibustion apparatus group was intervened by DZWJY-1 type electronic moxibustion apparatus, and the traditional moxibustion group received moxa stick moxibustion for treatment. Neixiyan (EX-LE 4), Dubi (ST 35), Xuehai (SP 10) and Liangqiu (ST 34) were selected for both groups and the treatment was conducted 3 times a week for a total of 12 times. The visual analog scale (VAS) and the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) scores were observed before treatment and after 6 and 12 sessions of treatment, respectively. Results: There were 4 dropout cases in the traditional moxibustion group. Therefore, this trial had 72 valid cases, including 38 cases in the electronic moxibustion apparatus group and 34 cases in the traditional moxibustion group, the differences in the baseline data between the two groups were statistically insignificant (P>0.05). After 6 and 12 sessions of treatment, the VAS scores decreased significantly with the increase of treatment sessions in both groups (all P<0.01), and the betweengroup differences were statistically insignificant at the same time points (both P>0.05). The pain intensity was evaluated using the weighted value of VAS score. The markedly effective rate was 47.4％ and the total effective rate was 89.5％ in the electronic moxibustion apparatus group, versus 50.0％ and 94.1％ in the traditional moxibustion group, and the betweengroup differences were statistically insignificant (both P>0.05). After 6 and 12 sessions of treatment, the total score and the component scores including pain, stiffness and difficulty moving in the WOMAC decreased significantly with the increase of treatment sessions in both groups (all P<0.01), and the between-group differences were statistically insignificant (all P>0.05). Conclusion: Electronic moxibustion apparatus and traditional moxibustion both are effective in reducing joint pain and improving joint function in KOA patients, and they are equivalent comparing the clinical efficacy.

Objective:To study on the differences of profile spectra among chemical components in Magnoliae Officinalis Cortex from different varieties and habitats,and to screen and identify the characteristic components affecting the quality difference of this herb. Method:The chromatogram data sets of Magnoliae Officinalis Cortex from different varieties and habitats were obtained by liquid chromatography-time-of-flight-mass spectrometry(LC-TOF-MS).Principal component analysis,partial least squares-discriminant analysis and cluster analysis were used to compare the differences in chemical profiles among Magnoliae Officinalis Cortex from different varieties and habitats,and adopted to screen out the characteristic chemical constituents that resulted in these differences and to perform mass spectrometry analysis and comparison. Result:Eleven characteristic peaks were identified by LC-TOF-MS chromatographic data and reported in the literature.The use of chemical profile could distinguish different habitats of Magnoliae Officinalis Cortex,but could not completely distinguish different varieties of this herb. Conclusion:LC-TOF-MS can easily and quickly study on the profile differences of chemical substances in Magnoliae Officinalis Cortex from different varieties and different habitats,the results of this study can provide a theoretical basis for the quality evaluation and pharmacodynamic material basis of this herb.

In forensic pathology, the estimation of postmortem interval （PMI） has always been a difficult issue, and there is still lack of effective methods to estimate PMI of corpses in water. Microbial biofilm refers to the microbial population attached to non-biological or biological surfaces by microorganisms during microbial growth, that has a three-dimensional structure, surrounded by extracellular polymers and matrix networks created by itself. A series of community succession phenomena of microorganisms occur during the occurrence and development of microbial population. The microbial community and its succession process of this kind of biofilm attached to the surface of a corpse in water may become a new basis for estimation of the PMI of corpses in water. This review elucidates on the concept, classification, research methods, and influencing factors of biofilm and analyzes its application prospects in PMI estimation of corpses in water, which would provide new ideas for the researches in this field.
Autopsy ; Biofilms ; Cadaver ; Drowning ; Forensic Pathology/methods* ; Humans ; Postmortem Changes ; Water