The Mechanism of miR-1294 Targeting SOX15 to Regulate Wnt/β-catenin Signaling Pathway and Promote the Proliferation of Acute Lymphoblastic Leukemia Cells in Children / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of abnormal miRNA expression on the proliferation of pediatric acute lymphoblastic leukemia (ALL) cells and its related mechanism.@*METHODS@#15 children with ALL and 15 healthy subjects were collected from the Second Affiliated Hospital of Hainan Medical University from July 2018 to March 2021. MiRNA sequencing was performed on their bone marrow cells, and validated using qRT-PCR. MiR-1294 and miR-1294-inhibitory molecule (miR-1294-inhibitor) were transfected into Nalm-6 cells, and the proliferation of Nalm-6 cells was detected by CCK-8 and colony formation assays. Western blot and ELISA were used to detect apoptosis of Nalm-6 cells. Biological prediction of miR-1294 was performed to find the target gene, which was verified by luciferase reporter assay. Si-SOX15 was transfected into Nalm-6 cells, Western blot was used to detect the expression of Wnt signaling pathway-related proteins and to verify the effect of si-SOX15 on the proliferation and apoptosis of Nalm-6 cells.@*RESULTS@#Compared with healthy subjects, 22 miRNAs were significantly upregulated in bone marrow cells of ALL patients, of which miR-1294 was the most significantly upregulated. In addition, the expression level of SOX15 gene was significantly reduced in bone marrow cells of ALL patients. Compared with the NC group, the miR-1294 group showed increased protein expression levels of Wnt3a and β-catenin, faster cell proliferation, and more colony-forming units, while caspase-3 protein expression level and cell apoptosis were reduced. Compared with the NC group, the miR-1294-inhibitor group showed reduced protein expression levels of Wnt3a and β-catenin, slower cell proliferation, and fewer colony-forming units, while caspase-3 protein expression level was increased and apoptosis rate was elevated. miR-1294 had a complementary base-pair with the 3'UTR region of SOX15 , and miR-1294 directly targeted SOX15 . The expression of miR-1294 was negatively correlated with SOX15 in ALL cells. Compared with the si-NC group, the si-SOX15 group showed increased protein expression levels of Wnt3a and β-catenin, accelerated cell proliferation, and decreased caspase-3 protein expression level and cell apoptosis rate.@*CONCLUSION@#MiR-1294 can target and inhibit SOX15 expression, thus activating the Wnt/β-Catenin signaling pathway to promote the proliferation of ALL cells, inhibit cell apoptosis, and ultimately affect the disease progression.

A new sesquiterpene glycoside from Osmanthus fragrans var. aurantiacus / 药学学报

The chemical constituents were isolated and purified by column chromatography and semi-preparative reversed-phase high performance liquid chromatography with silica gel, MCI and polyamide in order to study the chemical constituents of dried flowers of Osmanthus fragrans var. aurantiacus. Their structures were identified by the physical and chemical properties and one-dimensional nuclear magnetic resonance (1H-, 13C-NMR, DEPT), two-dimensional nuclear magnetic resonance (1H-1H COSY, non-decoupled HSQC, HSQC, HMBC), UV, IR and high resolution mass spectrometry data. One new compound (1) and five known compounds (2-6) were isolated from 95% ethanol extract of dried broccoli. They were identified as (9S)-9-hydroxymengastigm-5-en-4-one-9-O-primeveroside (1), oleanolic acid (2), forsythiaside (3), 2-(4-hydroxyphenethyl)-ethanol-(6-acetyl)-β-D-glucopyranoside (4), salidroside (5), and acteoside (6). Compounds (2-6) were isolated from this plant for the first time.

Quantitative study of nine main components in Yueju Tablets by HPLC coupled with wavelength switching and gradient elution method / 中草药

Objective: To establish HPLC coupled with wavelength switching and gradient elution method (HPLC-DVD) for simultaneous determination of nine main components (α-cyperone, genipin-1-β-D-gentiobioside, geniposide, crocin I, senkyunolide H, senkyunolide I, senkyunolide A, ligustilide, and atractylodin) in Yueju Tablets. Methods: The chromatographic separation was achieved on Zorbax Eclipse Plus C18 (250 mm × 4.6 mm, 5 μm) column with methanol-acetonitrile (2∶1) (A)-0.2% glacial acetic acid solution (B) as mobile phases for gradient elution, at the flow rate of 0.9 mL/min; The detection wavelength was set at 240 nm for α-cyperone, genipin-1-β-D-gentiobioside and geniposide, 440 nm for crocin I, 280 nm for senkyunolide H, senkyunolide I, senkyunolide A and ligustilide, and 340 nm for atractylodin. The volume of sample injection was 10 μL. Results: The nine active components were well separated and showed good linearity, such as α-cyperone 2.58—51.60 μg/mL (r = 0.999 4), genipin-1-β-D- gentiobioside 11.99—239.80 μg/mL (r = 0.999 9), geniposide 17.96—359.20 μg/mL (r = 0.999 6), crocin-I 3.98—79.60 μg/mL (r = 0.999 7), senkyunolide H 2.82—56.40 μg/mL (r = 0.999 9), senkyunolide I 2.38—47.60 μg/mL (r = 0.999 9), senkyunolide A 6.04—120.80 μg/mL(r = 0.999 5), ligustilide 7.98—159.60 μg/mL (r = 0.999 3), and atractylodin 6.51—130.20 μg/mL (r = 0.999 2). The precision was good, and RSD was not more than 1.22%. The repeatability was good, and RSD was not more than 1.75%. The stability was good in 18 h, and RSD was not more than 1.37%. The average recoveries and corresponding RSD values were 97.64% (0.98%), 99.09% (1.46%), 100.11% (1.03%), 97.87% (0.80%), 98.59% (1.19%), 96.89% (1.34%), 99.38% (0.58%), 98.50% (1.22%), and 99.71% (0.85%), respectively. The contents of 10 batches of α-cyperone, genipin-1-β-D-gentiobioside, geniposide, crocin I, senkyunolide, H, senkyunolide I, senkyunolide A, ligustilide and atractylodin were 0.282—0.344, 2.099—2.445, 3.628—4.225, 0.758—0.913, 0.241—0.286, 0.217—0.266, 1.077—1.291, 1.386—1.623, and 1.137—1.434 mg/tablet. Conclusion: HPLC coupled with wavelength switching and gradient elution method has been established for simultaneous determination of nine components in Yueju Tablets. The method is simple, quick, accurate, and it can be used for content determination and quality control of Yueju Tablets.

Chemical constituents from the stems of Gymnema sylvestre / 中国天然药物

AIM@#To study the chemical constituents of stems of Gymnema sylvestre (Retz.) Schult.@*METHODS@#Chromatographic techniques using silica gel, C18 reversed phase silica gel, and prep-HPLC were used. The structures were elucidated on the basis of MS and spectroscopic analysis (1D and 2D NMR), as well as chemical methods.@*RESULTS@#Seven compounds were isolated and their structures were elucidated as conduritol A (1), stigmasterol (2), lupeol (3), stigmasterol-3-O-β-D-glucoside (4), the sodium salt of 22α-hydroxy-longispinogenin-3-O-β-D-glucopyranosyl-(1→3)-β-D-glu-curono-pyranosyl-28-O-α-L-rhamnopyranoside (5), oleanolic acid-3-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (6), and the sodium salt of 22α-hydroxy-longispinogenin 3-O-β-D-glucuronopyranosyl-28-O-α-L-rhamnopyranoside (7). The inhibition activities of compounds 1, 5-7 on non-enzymatic glycation of protein in vitro were evaluated.@*CONCLUSION@#Compound 7 is a new triterpenoid saponin. It was shown that compounds 1, 5-7 have weak inhibition activities for non-enzymatic glycation of protein in vitro.

Relationship between hedgehog signaling pathway molecules and HPV16 infection in uterine cervical cancers / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the significance of sonic hedgehog (Shh), indian hedgehog (Ihh), smoothened (Smo) and patched (Ptch) expressions in uterine cervical lesions and their relationships with HPV type 16 infection.</p><p><b>METHODS</b>Totally 183 cases of cervical lesions, including 32 non-neoplastic cervix, 71 cervical intraepithelial neoplasia (28 CINI, 18 CINII, and 25 CINIII) and 80 squamous cell carcinomas (SCC) were selected from the Department of Pathology, Yanbian University Hospital, Yanbian Women Hospital, and Yanbian Tumor Hospital. Shh, Ihh, Ptch and Smo proteins expression were investigated by immunohistochemistry using tissue microarry platform, and the presence of HPV type 16 was detected by PCR method.</p><p><b>RESULTS</b>Immunohistochemical staining showed that the frequencies of Shh, Ihh, Ptch and Smo expression were rare in normal cervical epithelium, but were strongly expressed in cervical cancer and its precursor lesions (CINII/III) (P < 0.01, P < 0.01, P < 0.05, P < 0.05, respectively). In cervical cancer, the expression rate of Shh (95%) was higher than that of CIN (CINI to CINIII) (46.4%, 61.1%, 80.0%, respectively, P < 0.05). HPV16 was positive in 77.5% of SCC. In cervical cancer, the expression of Shh was related with HPV16 infection (P < 0.05), and the expression of Smo was correlated with lymph node metastasis (P < 0.05).</p><p><b>CONCLUSIONS</b>Shh, Ihh, Ptch, and Smo genes may play important roles in the development of cervical cancer. Detection of Hedgehog signaling pathway molecules seems helpful for the early diagnosis of cervical cancer and its precursor lesions, and are potentially therapeutic targets as well.</p>

Interference of Hepatocyte Growth-Promoting Factor on Cell Apoptosis in Kidney of Rats with Renal Ischemia Reperfusion Injury / 实用儿科临床杂志

Objective To explore the effects of hepatocyte growth-promoting factor(pHGF) on renal function and cell apoptosis in kidney of rats with renal ischemia reperfusion injury(IRI).Methods Thirty-two male Sprague-Dawley rats were divided into 4 groups:sham-operated control group(groupⅠ),renal ischemia reperfusion control group(groupⅡ),one experimental group injecting pHGF(50 mg/kg,intraabdominal injection) before renal IRI(group Ⅲ),and another experimental group injecting pHGF(50 mg/kg,intraabdominal injection) after renal IRI(group Ⅳ).The animals with renal IRI exposed to 45 min bilateral renal pedicle clamping.All ischemia reperfusion rats in group Ⅰ and Ⅱ were intraabdomially injected equal volume of physiological saline(0.8 mL) at the time when the rats in experimental groups were administered 50 mg/kg pHGF.Twelve hours after IRI,samples for serum and the left renal tissue of each animal were taken.The serum sample was used to detect expression of serum creatinine(Scr),and the renal tissue sample for evaluation of apoptosis.Results Compared with the level of Scr in groupⅠ(22.775?6.508) ?mol/L,Scr was markedly higher in groupⅡ(120.850?22.237) ?mol/L(P0.05).Conclusions The laboratory investigation suggests that pHGF might be an effective pharmacological agent against renal IRI according to the findings of the evaluated parameters,and protective effect by pHGF against renal IRI might involved in the mechanisms decreasing tubular cells apoptosis.It is likely that pHGF is a potential therapentic agent in clinical renal IRI circumstances.