Paris rugosa(Melanthiaceae) only grows in Yunnan province of China at present, and its chemical constituents have not been systematically studied. In this study, nine compounds, including one new compound pariposide G(1) and eight known compounds of cerin(2), stigmast-4-en-3-one(3), β-ecdysone(4), ophiopogonin C'(5), methyl protogracillin(6), gracillin(7), parissaponin H(8), and parisyunnanoside G(9), were isolated and identified from the ethanol extract of P. rugosa rhizomes by column chromatography methods and semi-preparative high-performance liquid chromatography(HPLC). Compounds 1-9 were isolated from this plant for the first time. The antibacterial and antifungal activities of all the compounds were evaluated. The results showed that ophiopogonin C' had strong inhibitory effects on Candida albicans [MIC_(90)=(4.68±0.01) μmol·L~(-1)] and the fluconazole-resistant strain of C. albicans [MIC_(90)=(4.66±0.02) μmol·L~(-1)].
Anti-Bacterial Agents ; Candida albicans ; China ; Liliaceae ; Melanthiaceae ; Rhizome

Abstract：Objective To improve the replication level of varicella⁃zoster virus（VZV）in human diploid cell line MRC⁃5 and increase the yield of VZV vaccine by reducing the expression of interferon（IFN）related genes via optimizing the cell line MRC⁃5. Methods Interferon receptor 1（IFNAR1）silenced MRC⁃5 cell line（MRC⁃5IFNAR1⁃）was constructed by CRISPR／Cas9 gene editing technology，which was determined for the relative expression of IFNAR1 mRNA，and for those of mRNA of IFN related genes IFNβ and OAS1 after VZV infection by qRT⁃PCR to evaluate the effect of gene silencing. Gene mutation sequences were further identified by sequencing of the silenced sites. The replication of VZV in MRC⁃5 and MRC⁃5IFNAR1⁃ cell lines was compared 168 h after VZV infection by using qRT⁃PCR and plaque formation unit（PFU）assay， to evaluate the effect of MRC⁃5IFNAR1⁃cell line on VZV replication. Results The growth status of MRC⁃5IFNAR1⁃ cell line wasconsistent with that of MRC ⁃ 5 cells，and the relative expression of IFNAR1 mRNA decreased by 73%；The relative expressions of IFNβ and OAS1 mRNA in MRC⁃5IFNAR1⁃ cell line were 61% and 90% lower than those in MRC⁃ 5 cells respectively after VZV infection；In addition，168 h after VZV infection，the level of DNA replication and the titer of VZV increased by 5. 7 folds and 4 folds respectively. Conclusion The successful establishment of MRC⁃5IFNAR1⁃ cell line may be a potential scheme to increase the yield of vaccines based on human diploid cells，and provided a reference for expanding production of VZV vaccine.

The common development of multi-disciplinary intersection is a hot spot in the research of acupuncture- moxibustion translational medicine. This article analyzes the current situation and reasons for slow development of acupuncture-moxibustion translational medicine, takes acupuncture-moxibustion for depressive disorder as an example, takes acupuncture and moxibustion literature, clinical evidence-based, biological mechanism and medical equipment research and development as the main line, expounds potential strategies to promote the development of acupuncture-moxibustion translational medicine under the background of multi-disciplinary intersection innovation, and discusses the future research direction of acupuncture-moxibustion translational medicine.
Translational Science, Biomedical

Objective:In general, patients with seropositive rheumatoid arthritis (RA) are considered to show an aggressive disease course. However, the relationship between the two subgroups in disease severity is controversial. Our study is aimed to compare the clinical characteristics and prognosis of double-seropositive and seronegative RA in China through a real-world large scale study.Methods:RA patients who met the 1987 American College of Rheumatology (ACR) classification criteria or the 2010 ACR/European Anti-Rheumatism Alliance RA classification criteria, and who attended the 10 hospitals across the country from September 2015 to January 2020, were enrolled. According to the serological status, patients were divided into 4 subgroups [rheumatoid factor (RF)(-) anti-cyclic citrullinated peptide (CCP) antibody (-), RF(+), RF(+) anti-CCP antibody(+), anti-CCP antibody(+)] and compared the disease characteristics and treatment response. One-way analysis of variance was used for measurement data that conformed to normal distribution, Kruskal-Wallis H test was used for measurement data that did not conform to normal distribution; paired t test was used for comparison before and after treatment within the group if the data was normally distributed else paired rank sum test was used; χ2 test was used for count data. Results:① A total of 2 461 patients were included, including 1 813 RF(+) anti-CCP antibody(+) patients (73.67%), 129 RF(+) patients (5.24%), 245 RF(-) anti-CCP antibody(-) patients (9.96%), 74 anti-CCP antibody(+) patients (11.13%). ② Regardless of the CCP status, RF(+) patients had an early age of onset [RF(-) anti-CCP antibody(-) (51±14) years old, anti-CCP antibody(+) (50±15) years old, RF(+) anti-CCP antibody(+) (48±14) years old, RF(+)(48±13) years old, F=3.003, P=0.029], longer disease duration [RF(-) anti-CCP antibody(-) 50 (20, 126) months, anti-CCP antibody(+) 60(24, 150) months, RF(+) anti-CCP antibody(+) 89(35, 179) months, RF(+) 83(25, 160) months, H=22.001, P<0.01], more joint swelling counts (SJC) [RF(-) anti-CCP antibody(-) 2(0, 6), Anti-CCP antibody(+) 2(0, 5), RF(+) anti-CCP antibody(+) 2(0, 7), RF(+) 2(0, 6), H=8.939, P=0.03] and tender joint counts (TJC) [RF(-) anti-CCP antibody(-) 3(0, 8), anti-CCP antibody(+) 2(0, 6), RF(+) anti-CCP antibody(+) 3(1, 9), RF(+) 2(0, 8), H=11.341, P=0.01] and the morning stiff time was longer [RF(-) anti-CCP antibody(-) 30(0, 60) min, anti-CCP antibody(+) 20(0, 60) min, RF(+) anti-CCP antibody(+) 30(10, 60) min, RF(+) 30(10, 60) min, H=13.32, P<0.01]; ESR [RF(-) anti-CCP antibody(-) 17(9, 38) mm/1 h, anti-CCP antibody(+) 20(10, 35) mm/1 h, RF(+) anti-CCP antibody(+) 26(14, 45) mm/1 h, RF(+) 28(14, 50) mm/1 h, H=37.084, P<0.01] and CRP [RF(-) anti-CCP antibody(-) 2.3 (0.8, 15.9) mm/L, Anti-CCP antibody(+) 2.7(0.7, 12.1) mm/L, RF(+) anti-CCP antibody(+) 5.2(1.3, 17.2) mm/L, RF (+) 5.2(0.9, 16.2) mm/L, H=22.141, P<0.01] of the RF(+)patients were significantly higher than RF(-) patients, and RF(+) patients had higher disease severity(DAS28-ESR) [RF(-) anti-CCP antibody(-) (4.0±1.8), anti-CCP antibody(+) (3.8±1.6), RF(+) anti-CCP antibody(+) (4.3±1.8), RF(+) (4.1±1.7), F=7.269, P<0.01]. ③ The RF(+) anti-CCP antibody(+) patients were divided into 4 subgroups, and it was found that RF-H anti-CCP antibody-L patients had higher disease severity [RF-H anti-CCP antibody-H 4.3(2.9, 5.6), RF-L anti-CCP antibody-L 4.5(3.0, 5.7), RF-H anti-CCP antibody-L 4.9(3.1, 6.2), RF-L anti-CCP antibody-H 2.8(1.8, 3.9), H=20.374, P<0.01]. ④ After 3-month follow up, the clinical characteristics of the four groups were improved, but there was no significant difference in the improvement of the four groups, indicating that the RF and anti-CCP antibody status did not affect the remission within 3 months. Conclusion:Among RA patients, the disease activity of RA patients is closely related to RF and the RF(+) patients have more severe disease than RF(-) patients. Patients with higher RF titer also have more severe disease than that of patients with low RF titer. After 3 months of medication treatment, the antibody status does not affect the disease remission rate.

Chaperone-mediated autophagy (CMA) is a lysosome-dependent selective degradation pathway implicated in the pathogenesis of cancer and neurodegenerative diseases. However, the mechanisms that regulate CMA are not fully understood. Here, using unbiased drug screening approaches, we discover Metformin, a drug that is commonly the first medication prescribed for type 2 diabetes, can induce CMA. We delineate the mechanism of CMA induction by Metformin to be via activation of TAK1-IKKα/β signaling that leads to phosphorylation of Ser85 of the key mediator of CMA, Hsc70, and its activation. Notably, we find that amyloid-beta precursor protein (APP) is a CMA substrate and that it binds to Hsc70 in an IKKα/β-dependent manner. The inhibition of CMA-mediated degradation of APP enhances its cytotoxicity. Importantly, we find that in the APP/PS1 mouse model of Alzheimer's disease (AD), activation of CMA by Hsc70 overexpression or Metformin potently reduces the accumulated brain Aβ plaque levels and reverses the molecular and behavioral AD phenotypes. Our study elucidates a novel mechanism of CMA regulation via Metformin-TAK1-IKKα/β-Hsc70 signaling and suggests Metformin as a new activator of CMA for diseases, such as AD, where such therapeutic intervention could be beneficial.

【Objective】 To predict the functional mechanism and clinical significance of highly expressed maternal embryonic leucine zipper kinase(MELK) in lung adenocarcinoma and lung squamous cell carcinoma by bioinformatics. 【Methods】 The chip data set GSE7670 was obtained from the Gene Expression Omnibus(GEO) database and combined with the TCGA lung adenocarcinoma(LUAD) and lung squamous cell carcinoma(LUSC) database for differential analysis, and to screen out differential kinases(P < 0.05). GO biological function enrichment and KEGG pathway analysis of these kinases were performed. Based on String, a protein interaction network was obtained. MCODE plug-in of Cytoscape was applied to find key node proteins and then MELK kinase protein was picked out. MELK expression was analyzed through Oncomine database. Clinical information from TCGA database was obtained, and the relationship between MELK expression and clinicopathological characteristics, prognosis of LUAD and LUSC was analyzed. GeneMANIA was used to predict the function of MELK, and then to verify related co-expressed genes with GEPIA2. 【Results】 A total of 159 differential kinases were screened out, which were closely related to cell proliferation, apoptosis and various signal pathways. Twenty-one important genes were identified by analyzing protein interaction networks. MELK level was higher in male, young or advanced clinical stage lung adenocarcinoma and lung squamous cell carcinoma. The patients with high MELK levels had a shorter progression-free survival(PFS) and overall survival(OS) in LUAD, but there was no significant difference in LUSC. MELK was involved in G2/M transition possibly and was co-expressed with proliferation-related genes (MKI67, PCNA, CCNB1, MCM2, and TOP2A) in LUAD and LUSC. 【Conclusions】 MELK is highly expressed in LUAD and LUSC. MELK may be involved in the occurrence and development of LUAD by promoting cell mitosis and G2/M transition. MELK could be a new biomarker for LUAD.

Adolescent ; Adult ; Female ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid ; genetics ; Male ; Oncogene Proteins, Fusion ; genetics ; Retrospective Studies

OBJECTIVETo investigate the appropriate extubation time and treatment of late complications after early tracheotomy in patients with moderate or severe inhalation injury.

METHODSOne hundred and fifty patients (105 males and 45 females) with inhalation injury were admitted to our hospital from January 2000 to January 2009. Among them, 109 out of 129 cases with moderate inhalation injury received early tracheotomy, and all 21 cases with severe inhalation injury received early tracheotomy. Data were collected for analysis as follows: (1) incidence of re-intubation due to suffocation and pneumonia incidence after extubation within 2 weeks or after 2 weeks post inhalation injury (PII), and mortality rate within the first week after injury were recorded. (2) Conservative treatments including expectorant, oral antibiotics, and absolute bedrest were recommended for patients who had severe cough, hoarseness or poor pulmonary function after late extubation and closure of tracheostomy wound. Fiberoptic bronchoscopy findings (tracheostenosis degree, granuloma formation rate, vocal cord paralysis rate) and pulmonary function index (FEV(1)) data were collected and analyzed in 30 cases with moderate inhalation injury and 10 cases with severe inhalation injury within 3 months after injury for follow-up. Data were processed with t test or chi-square test.

RESULTSThere was no obvious difference in the rate of re-intubation after extubation in patients with moderate inhalation injury between those done within 2 weeks PII (15/70, 21.4%) and those done after 2 weeks PII (2/25, 8.0%) (χ(2) = 1.52, P > 0.05). Pneumonia incidence in patients of moderate inhalation injury with extubation within 2 weeks PII (21/70, 30.0%) was lower than those with extubation after 2 weeks PII (15/25, 60.0%) (χ(2) = 7.04, P < 0.05). Levels of above-mentioned indexes in patients with severe inhalation injury extubated in different stages were similar to those of patients with moderate inhalation injury. Within the first week after injury, mortality rate of patients with severe inhalation injury was higher than that of patients with moderate inhalation injury (χ(2) = 11.90, P < 0.05). During follow-up, tracheostenosis rate in patients with moderate or severe inhalation injury was 100.0%; granuloma formation rate and vocal cord paralysis rate in patients with severe inhalation injury were higher than those of patients with moderate inhalation injury (with χ(2) value respectively 4.59, 13.47, P values all below 0.05). The FEV(1) value of patients with moderate inhalation injury in the 1st, 2nd, 3rd month after injury was respectively higher than that of patients with severe inhalation injury (with t value respectively 5.48, 12.10, 6.25, P values all below 0.05). The values recovered to normal level in the 3rd month after injury.

CONCLUSIONSExtubation time of tracheotomy for patients with moderate or severe inhalation injury within 2 weeks or after 2 weeks PII has its own advantage and disadvantage, and it should be performed according to specific conditions of each patient. Conservative treatment is optional for late complications of respiratory system.

Adolescent ; Adult ; Aged ; Burns, Inhalation ; surgery ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Intubation, Intratracheal ; adverse effects ; Male ; Middle Aged ; Postoperative Complications ; Tracheotomy ; adverse effects ; Young Adult

OBJECTIVETo identify the HBV genotype-specific tag sequence.

METHODSThe large S region sequences from 930 HBV genomes were aligned to identify the genotype-specific tag sequences. PCR was used to check the genotyping effect of these tags.

RESULTSTwo tag sequences, sequence between 149-169 and sequence between 461-483, were identified in the large S region. Using primers specific to these tag sequences, the genotype of HBV can be specifically identified.

CONCLUSIONThese tag sequences can be used for HBV genotyping.

Base Sequence ; DNA Primers ; Gene Library ; Genes, Viral ; Genotype ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Protein Precursors ; genetics ; Sequence Analysis, DNA