OBJECTIVETo establish the chromatographic fingerprints for the anti-tumor flavonoids of Caulis spatholobi (SSCE). It could used to reflect the chemical information in this part comprehensively, and identify the chemical consitituents preliminarily.

METHODThe HPLC-DAD analysis method was performed on the column Kromasil 100-5PHENYL (4.6 mm x 250 mm, 5 microm). The mobile phase was water (0.5% acetic acid)- methanol in gradient elution and the detection wavelength was 254 nm.

RESULTThe chromatographic fingerprint of SSCE was established, which showed 16 characteristic peaks from 10 batches of medicinal materials. Among them, the peaks 1, 3, 4, 5, 8, 9, 10, 12, 13, and 16 were identified 3,4-dihodroxybenzoic acid, 4-Hydroxybenzoic Acid, epicatechin, puerarin, daidzein, liquiritigenin, calycosin, genistein, formononetin, and prunetin, respectively.

CONCLUSIONThe method is convenient, reproducibility and stability. It can used for quality control of the anti-tumor flavonoids of C. spatholobi (SSCE).

Antineoplastic Agents ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; chemistry ; Fabaceae ; chemistry ; metabolism ; Flavonoids ; analysis ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results

Objective To compare the clinical effect of Ginkgo biloba extract gel (Ginkgo biloba extract,EGB) and minocycline hydrochloride (Periocline) on periodontitis and their inhibition on putative periodontal pathogens.Methods Thirty patients with moderate-to-severe periodontitis were selected.The patients were divided into an experimental group and a positive control group (minocycline hydrochloride) Supragingival and subgingival scaling were performed on all patients.Subgingival plaque samples were collected before treatment,1 week,2 months and 4 months after treatment.The four major periodontal pathogens Porphyromonas gingivalis (Pg),Bacteroides forsythus (Bf),Prevotella intermedia (Pi),Treponema denticola(Td) were detected by polymerase chain reaction.Clinical indexes plaque index (PLI),bleeding index(BI) and probing depth(PD),attachment loss (AL) were examined before treatment,3 months and 6 months after treatment.The results were statistically analyzed.Results The detection rates of the 4 periodontal pathogens were Td (83.3％),Tf(95.0％),Pi (80.0％),Pg(81.7％) in experimental group and Td (83.3 ％),Tf(95.0％),Pi (80.0％),Pg (81.7％) in control group before treatment.The detection rates in experimental group were not significantly different with those in control group after treatment,except for the detection rate of Pg 1 week after treatment (P ＜ 0.01,the detection of Pg was 56.7％ in experimental group and 53.3％ in control group 1 week after treatment).The PLI and BI were not significantly different between experimental group and control group after treatment (P ＞ 0.05).The difference was statistically significant at 6 months after treatment [(3.5 ± O.5) mm for experimental group and (3.2 ± 0.4) mm for control group,P =0.00].The mean of AL decreased with time.The difference was statistically significant at 6 months after treatment[(4.5 ±0.4) mm for experimental group and (4.3 ±0.4) mm for control group at 6 months,P =0.01].Conclusions The inhibition effects of EGB and minocycline hydrochloride were comparable for major periodontal pathogens within short term.

Objective To explore the effect of nicotinamide transmethylase on intracellular reactive oxygen species(ROS)in iron death of hepatocellular carcinoma cells and its mechanism.Methods Methyl nicotinamide(MNA)expression in cells was detected using a tandem liquid chromatography-mass spectrometry.The average fluorescence intensity of ROS and lipid peroxidation was measured using a flow cytometer.Western blot was used to detect changes in the expression of human liver cancer cells(SK-Hep-1,Hep3B).Forty patients with primary hepatocellular carcinoma who received treatment in our hospital from March 2019 to February 2020 were selected as the study subjects,and their adjacent tissue samples and liver cancer tissue samples were collected.Immunohistochemical methods were used to detect the levels of nicotinamide N-methyltransferase(NNMT)and ROS in adjacent and liver cancer tissues.CCK-8 method was used to detect the survival activity of cells with different iron concentrations.Results The MNA levels in the liver cancer tissue group were higher than those in the adjacent tissue group(P<0.05).Compared with the adjacent tissue group,the average fluorescence intensity expression of ROS in the liver cancer tissue group increased,while the average fluorescence intensity expression of lipid peroxidation decreased(P<0.05).Compared with the adjacent tissue group,the expression levels of SK Hep-1 and Hep3B cells in the liver cancer tissue group increased(P<0.05).Compared with the control group,NNMT groups 2,10,20,and 25 μmol/L The cell survival activity level increased(P<0.05);Compared with the NNMT group,the iron inhibition group had different iron concentrations(2μmol/L,10μmol/L,20μmol/L,25μmol/L.The expression of cell viability decreased(P<0.05).Conclusion ROS mediated by nicotinamide methyltransferase can be guided to produce ROS and energy disorders,leading to increased tumor cell death.

Spermatogonial stem cells (SSCs) have great applications in both reproductive and regenerative medicine. Primates including monkeys are very similar to humans with regard to physiology and pathology. Nevertheless, little is known about the isolation, the characteristics, and the culture of primate SSCs. This study was designed to identify, isolate, and culture monkey SSCs. Immunocytochemistry was used to identify markers for monkey SSCs. Glial cell line-derived neurotrophic factor family receptor alpha-1 (GFRA1)-enriched spermatogonia were isolated from monkeys, namely Macaca fascicularis (M. fascicularis), by two-step enzymatic digestion and magnetic-activated cell sorting, and they were cultured on precoated plates in the conditioned medium. Reverse transcription-polymerase chain reaction (RT-PCR), immunocytochemistry, and RNA sequencing were used to compare phenotype and transcriptomes in GFRA1-enriched spermatogonia between 0 day and 14 days of culture, and xenotransplantation was performed to evaluate the function of GFRA1-enriched spermatogonia. SSCs shared some phenotypes with rodent and human SSCs. GFRA1-enriched spermatogonia with high purity and viability were isolated from M. fascicularis testes. The freshly isolated cells expressed numerous markers for rodent SSCs, and they were cultured for 14 days. The expression of numerous SSC markers was maintained during the cultivation of GFRA1-enriched spermatogonia. RNA sequencing reflected a 97.3% similarity in global gene profiles between 0 day and 14 days of culture. The xenotransplantation assay indicated that the GFRA1-enriched spermatogonia formed colonies and proliferated in vivo in the recipient c-Kit