Cardiac glycosides from Streblus asper with potential antiviral activity.

Ten undescribed cardiac glycosides, strasperosides A-J, together with twelve known analogues, were isolated from Streblus asper Lour. Their structures were elucidated on the basis of spectroscopic analysis, electronic circular dichroism data, and chemical methods. These cardiac glycosides showed diversity in steroid skeleton and sugar moiety. Strasperosides A and B are a pair of unusual stereoisomers featuring different orientation of the lactone motif. Ten cardiac glycosides demonstrated potent antiviral effects on HSV-1 in vitro with the IC50 values from 0.19 ± 0.08 to 1.03 ± 0.25 µM and the therapeutic indices from 66.61 ± 5.08 to 326.75 ± 11.75.

Reliability of Computing van der Waals Bond Lengths of Some Rare Gas Diatomics.

When the bond lengths of 11 molecules containing van der Waals bonds are optimized by 572 methods and 20 basis sets, it is found that the best mean absolute deviations (MADs) of density-functional theory (DFT) methods are 0.005 Å (shown by APFD/6-311++G**), 0.007 Å (B2PLYPD3(Full)/aug-cc-pVQZ), and 0.010 Å (revDSDPBEP86/aug-cc-pVQZ), while the best MADs of ab initio methods are 0.008 Å (BD(T)/aug-cc-pVTZ) and 0.016 Å (MP4/aug-cc-pVQZ). Moreover, the best MADs calculated by 54 selected methods in combination with 60 other basis sets (such as 6-311++G, 6-31++G(3d'f,3p'd), and UGBS1V++) are not better. Therefore, these bond lengths can be calculated with extremely high accuracy by some special methods and basis sets, and CCSD(T) is also not as good as expected because its best MAD is only 0.023 Å (CCSD(T)/aug-cc-pVQZ).

Reliability of computed molecular structures.

When the structures of 1342 molecules are optimized by 30 methods and 7 basis sets, there appear 289 (21.54%) problematic molecules and 112 (8.35%) failed ones. When 278 problematic molecules are compared, the best methods are BHandH and LC-wPBE, while B97D, BP86, HFS, VSXC, and HCTH are very unreliable. When 179 problematic molecules are computed with larger basis sets, the smallest mean absolute deviation (MAD) of bond angle (2.3°) is shown by QCISD(T)/cc-pVTZ, while the smallest MAD of bond length (0.021 Å), the best SUM1 (4.9 unit), and the best SUM2 (2.4 unit) are shown by DSDPBEP86(Full), DSDPBEP86, PBE1PBE-D3, MP2, and MP2(Full) in combination with aug-cc-pVQZ, cc-pVQZ, Def2QZVP, Def2TZVPP, and/or 6-311++G(3df,3pd). Very large basis sets, for example, larger than cc-pVTZ usually have to be used to obtain very good structures and the performances of many density-functional theory methods are encouraging. The best results may be the limit of modern computational chemistry.

[Systematic review and trail sequential analysis of preparation of Xiakucao for Hashimoto's thyroiditis].

To systemically evaluate the clinical efficacy and safety of oral preparation of Xiakucao with levothyroxine(LT4) on Hashimoto's thyroiditis(HT), so as to provide the evidence for its clinical application in the future. All the included studies were retrieved from four Chinese databases and three English databases from their inception to December 2019. ROB assessment tool of cochrane system and the evidence classification recommended by GRADE were used to evaluate the quality of evidences in all included studies. RevMan 5.3 was used for Meta-analysis of the outcomes. Software TSA 0.9(trail sequential analysis) was used to estimate the sample size for Meta-analysis. The results showed that 11 randomized controlled trials and totaling 1 215 patients were included. Preparation of Xiakucao combined with LT4 was adopted as intervention in experimental group, while patients in control group were treated with LT4 alone. Meta-analysis results showed that as compared with control group, the rate of total efficacy in experimental group was significant improved, including improvement of thyroid function and thyroid autoantibodies, shrinkage of thyroid gland and nodule, and improvement of clinical symptoms such as fatigue and cold intolerance(RR=1.15, 95%CI[1.09, 1.21]). The experimental group significantly decreased the serum level of thyroperoxidase antibody TPO-Ab(SMD=-0.91, 95%CI[-1.40,-0.41]), and reduced the size of left thyroid lobe(MD=-1.46, 95%CI[-1.82,-1.11]), right thyroid lobe(MD=-1.45, 95%CI[-1.96,-0.94]) and isthmus of thyroid gland(MD=-1.08, 95%CI[-1.20,-0.95]). After evaluation based on GRADEpro, the results showed that the evidence quality of all included studies was low or very low. The result of TSA showed that the cumulative sample size had reached the expected value. However, the pooled results may be affected by one study with high bias risk, with not so high effect intensity of evidences. From this review, we can see that in treatment of HT, intervention of preparation of Xiakucao combined with LT4 has advantages on improvement of clinical efficiency, decreasing serum level of TPO-Ab and shrinkage of thyroid gland. However, due to the quality of evidence, more rigorously designed and high-quality trials are needed in the future to verify the clinical efficacy and safety of preparation of Xiakucao in treating HT.

Low CADM2 expression predicts high recurrence risk of hepatocellular carcinoma patients after hepatectomy.

PURPOSE: To investigate the expression and clinical significance of CADM2 in hepatocellular carcinomas (HCC). METHODS: The level of expression of CADM2 mRNA was assessed in frozen tumor specimens and adjacent noncancerous tissues from 30 HCC patients by real-time PCR. The protein level was determined by immunohistochemistry on a tissue microarray containing tumor and adjacent noncancerous tissues from 234 HCC patients. Clinicopathological characteristics associated analysis was performed through SPSS18 . RESULTS: CADM2 was strikingly down regulated in HCC. CADM2 expression was associated with differentiation (P = 0.000), serum alpha-fetoprotein (P = 0.003), vascular invasion (P = 0.001), and hepatitis B surface antigen (HBsAg, P = 0.038). Furthermore, patients with low CADM2 expression had significantly poorer recurrence-free survival (RFS) (40.8 and 34.2 % vs. 56.3 and 50.1 % in 3- and 5-year RFS, respectively, P = 0.005). Subgroup analysis revealed that the difference in RFS between groups with low- and high-CADM2 expression still existed among patients belonging to stage 0 or A of BCLC staging system (P = 0.008), patients with tumor ≤5 cm in size (P = 0.013), and alpha-fetoprotein-negative patients (P = 0.003). Moreover, low expression was more frequently observed in the early recurrence group (within 2 years, P = 0.007). Further multivariate Cox regression analysis indicated that CADM2 expression level, tumor size, tumor number, vascular invasion, HBsAg were independent risk factors for HCC recurrence. CONCLUSION: CADM2 serves as a novel predictor of RFS in HCC patients after curative resection.

Relevant factors on the degree of anterior uveitis in patients with ankylosing spondylitis / 国际眼科杂志(Guoji Yanke Zazhi)

AlM: To investigate the association between the degree of anterioruveitis and related factors including inflammatory markers as well as sacroiliac joint imaging in patients with ankylosing spondylitis ( AS) . METHODS: Anterior changes evaluated by slit lamp, erythrocyte sedimentation rate ( ESR ) , C - reactive protein ( CRP ) and magnetic resonance imaging of 55 cases with AS associateduveitis were retrospectively analyzed. A modified endotoxin-induced uveitis ( ElU ) clinical standard was used for uveitis grading. SPARCC sacroiliac scoring was used to evaluate bone edema of sacroiliac joint. The correlation between the degree of uveitis and sacroiliitis was assessed. RESULTS: ln the 55 patients with AS, ElU grading scored 2-10, and SPARCC index scored 0-22. Further analysis showed that the severity of uveitis was significantly correlated with ESR (r=0. 869, PCONCLUSlON: Local autoimmunity of uveitis and sacroiliac joint inflammation with subsequent bone formation in AS might be mutually independent processes.

Hepatitis C virus single-stranded RNA induces innate immunity via Toll-like receptor 7.

BACKGROUND/AIMS: Innate immune responses to HCV infection are triggered through host recognition of pathogen-associated molecular patterns. Interferons are critical for the protection against HCV infection. However, the pathways linking virus recognition to IFN induction remain poorly understood. METHODS: Immune cells and Huh-7 cells were infected with HCV cell culture (HCVcc) or transfected with HCV-derived immunostimulatory RNA oligonucleotides (ORNs), and immune activation was assessed. RESULTS: We found that HCVcc suppressed immune responses because the HCVcc protein impaired the PBMC and pDC responses. However, HCVcc genomic RNA had an immunostimulatory effect. HCV encodes G/U-rich ssRNA TLR7 ligands that significantly activate innate immunity, and induced IFN-alpha production. Moreover, HCV-derived ORNs also activated IRF7 and NF-kappaB in Huh-7 cells. In particular, the HCV 3'-UTR strongly induced cytokine production. Different lengths of polyuridine tract in the 3'-UTR of different HCV strains induced IFN-alpha production. These data demonstrate that the HCV-specific G/U fragment is a motif sequence, and is recognized by TLR7 as a PAMP. The requirement for TLR7 to recognize HCV RNA was confirmed using specific inhibitors, RNAi and by TLR7overexpression. CONCLUSION: These results provide an insight into the development of immune adjuvant for vaccines and for the production of new antiviral drugs.

Identification of the proteomic changes in Synechocystis sp. PCC 6803 following prolonged UV-B irradiation.

The diversified physiological responses in cyanobacteria under ultraviolet-B (UV-B) radiation have been broadly researched. The changes in the metabolic control mechanisms hidden behind these physiological traits still need to be further investigated. This research attempts to identify some of the internal mechanisms of several stressful phenotypes such as a decreased growth rate, an impaired photosystem, and the degradation of photosynthetic pigments. Different expression levels of proteins in the cytoplasm of Synechocystis sp. PCC 6803 under short-term and long-term UV-B stress were investigated by using a comparative proteomic approach. One hundred and twelve differentially expressed protein spots were identified by mass spectrometry to match 75 diverse protein species. They mainly focus on amino acid biosynthesis, photosynthesis and respiration, energy metabolism, protein biosynthesis, cell defence, and other functional groups. By focusing on these areas, the study reveals the correlation between UV-B stress-responsive proteins and the physiological changes listed above. The research, showing that short-term response-proteins are quite different from long-term response-proteins, helps to identify the change in homeostatic mechanisms in Synechocystis sp. PCC 6803. Related putative functions of these proteins and the physiological responses of cyanobacteria under UV-B stress, a UV-B responsive protein network in Synechocystis sp. PCC 6803 under long-term stress was successfully produced. Such a protein network helps to increase our understanding of the comprehensive functional network cyanobacteria use to adapt to UV-B stress. In addition, 30 novel proteins not previously found related to UV-B stress were identified. This opens up new areas for exploration to identify the response to UV-B stress in cyanobacteria.

Combined DNA vaccination against three animal viruses elicits decreased immunogenicity of a single plasmid in mice.

In order to investigate whether combined DNA vaccines are an ideal way to combine antigens in a single vaccine formulation, we immunized mice with three plasmids (pVSG, pVgD and pVE2), respectively, encoding the antigen of foot-and-mouth disease virus (FMDV), pseudorabies virus (PRV) and classic swine fever virus (CSFV), either alone or in a combined vaccine regimen. We also investigated the immune responses induced by a series of mixtures in which three plasmids were mixed in pairs. Then we further immunized mice with three different plasmids in separate sites and preformed an adoptive transfer experiment. While being given alone, each of the vaccine plasmids induced significant virus-specific antibody responses and splenocytes proliferative activity. But reduced immunogenicity of the pVSG plasmid was found in combined DNA vaccination, no matter whether it was injected in a single or a separate site. Removal single plasmid (pVgD or pVE2) from combined DNA vaccine led to significant increase in the immunogenicity of the pVSG plasmid (P<0.05). And the induction of immune suppression was not mediated by suppressor T cells, as demonstrated by an adoptive transfer experiment. Furthermore, by boosting with whole virus protein of FMDV, mice primed with either pVSG alone or combined DNA vaccine produced statistically significant increase in the FMDV-specific antibody titers (P<0.05). But after boosting, FMDV-specific splenocytes proliferative activity of mice primed with combined DNA vaccine was even lower than that of mice primed with pVSG alone (P<0.05). Taken together, this study reflected the immunogenicity of a single plasmid may be decreased in combined DNA immunization strategy, which still needs to be carefully evaluated before practical application.