OBJECTIVE: To study the distribution and phylogenesis of endophytic fungi in the stems and leaves of Desmos chinensis. METHODS: Permanent paraffin-cut section, optical microscope photography, and histochemistry were used to observe the distribution of endophytic fungi in the stems and leaves of Desmos chinensis. Plate isolation method was used to separate and cultivate the endophytic fungi. The genomic DNA of endophytic fungi were extracted, then the rDNA ITS region was amplified using polymerase chain reaction (PCR). The PCR products were digested by Hae III and Hha I restriction enzymes to determine the species and genotypes, and sequenced according to the digestion genotyping results. Nucleotide sequences of the rDNA ITS of endophytic fungi were used for the identification and phylogenetic analysis. RESULTS: Endophytic fungi mainly existed in the parenchyma cells of the phloem in the stems, and in the parenchyma cells, palisade tissue or sponge tissue of the leaves. Fourteen strains of endophytic fungi were isolated from the stems and leaves. Four different genotypes and their sequences were obtained from RFLP analysis and sequencing. These strains were identified as Guignardia mangiferae, Colletotrichum gloeosporioides, and Phomopsis spp. The dominant group of those endophytic strains was Phomopsis, represented by 8 strains and accounting for 57.1% of all. CONCLUSION: The endophytic fungi distribute in both stems and leaves of Desmos chinensis without obvious tissue specificity. The result of the phylogenetic analyses with rDNA ITS sequences reveals the phylogenetic relationships of endophytic fungi in Desmos chinensis.

Objective:To investigate the liposoluble constituents in fruit of Cnidium monnieri. Methods:The liposoluble constitu-ents in fruit of C. monnieri were extracted by Soxet extraction. The extracted liposoluble constituents were methyl esterified and then analyzed by GC-MS for the first time. Results:Thirty-nine compounds (85. 33%) were identified from the liposoluble constituents in fruit of C. monnieri. Phytol (15. 98%), hexadecanoic acid (12. 73%), 9,12,15-octadecatrienoic acid (11. 02%), 9-octadecenoic acid (6. 33%) and 9,12-octadecadienoic acid (4. 77%) were the main constituents of the liposoluble constituents in fruit of C. mon-nieri. Conclusion:Phytol, hexadecanoic acid, 9,12,15-octadecatrienoic acid and 9-octadecenoic acid are the active ingredients in fruit of C. monnieri.

Objective: To study the prescription and preparation technology of tea tree oil (TTO) microemulsion gel. The quality and the stability were evaluated. Methods: The prescription and preparation technology were selected and optimized through the compatibility test and the pseudo-ternary phase diagram. TTO microemulsion gel was prepared by adding gelatin. The appearance, pH value, viscosity, moisture rate, and the drug concentration were evaluated. Results: The prescription composition of TTO microemulsion gel was TTO (0.6%), Cremophor RH-40 (1.2%), PEG 400 (0.2%), carbopol-980 (0.2%), glycerol (2%), with distilled water adding to 50 g. The optimum formulation exhibited clear and transparent, uniform exquisite, moderate viscosity, and well spreadable, with the particle diameter of (38.38 ± 2.30) nm, zeta potential of (-56.00 ± 5.82) mV, pH value of 5.52 ± 0.01, viscosity of (48 834 ± 5) Pa·s, moisture rate of (96.74 ± 0.52)%, and the drug-loaded of (5.79 ± 0.03) mg/g. The result of heat and cold resistance test showed that the preparation was needed to be stored at low-temperature. Conclusion: The preparation of TTO microemulsion gel is simple, corresponding to the main index of gelata for topical drug delivery preparation and offering the basis for further research and development.

Objective To study the effects of atractylodes I, II, and III against rotavirus in vitro and in vivo. Methods An in vitro study model was established using Caco-2 cells. The cytopathic effect (CPE) and MTT staining were used to determine the toxicity of atractylenolide I, II, and III to cells for the inhibition of rotavirus biosynthesis, direct inactivation of rotavirus, and antiviral adsorption, with ribavirin as a positive drug. With half of the therapeutic concentration (EC50) and half of the cytotoxic concentration (TC50), the treatment index TI value was obtained and used as the evaluation index. An RV-infected model of suckling diarrhea was established in vivo to observe the signs and symptoms of the suckling mice, and the in vivo anti-rotavirus effect was preliminarily determined according to the diarrhea score and the weight gain. Results In vitro studies found that atractylenolide III had the direct inactivation effect on rotavirus with TI value of 8; atractylodes III medium-dose group has the best anti-rotavirus effect in vivo. Conclusion Atractylodes III, the main active component of Atractylodes macrocephala, has significant anti-rotavirus effect in vitro and in vivo; Atractylenolide III mainly works by directly inactivating rotavirus in vitro.

Objective To study the prescription and preparation technology of tea tree oil gel, and evaluate its anti-inflammatory efficacy, antibacterial effect and the irritation. Methods The tea tree oil gel was prepared using the carbomer-940 as gel matrix, Cremophor RH-40 and 1,2-propylene glycol as solvents. The appearance characters, pH value, viscosity, moisture retention, drug content, and the stability were observed. The anti-inflammatory efficacy, the antibacterial effect and the irritation of tea tree oil gel were evaluated. Results The prescription of tea tree oil gel was selected as following tea tree oil (1.0%), Cremophor RH-40 (5.0%), 1,2-propylene glycol (5.0%), Carbomer-940 (0.6%), glycerol (8.0%), with distilled water 100 g, adjusting pH to 5.0 by triethanolamine. The gel exhibited transparent, well uniformity, appropriate viscosity and fine coating expansion performance, with pH value of 5.52 ± 0.03, viscosity at (48 782 ± 25) mPa•s, the moisture retaining rate of (93.32 ± 0.38)% for 24 h test, containing tea tree oil of (9.55 ± 0.10) mg/g. The inhibition rate of tea tree oil gel on the mouse auricle swelling was 46.15%, which was significantly different as compared to the negative control group (P < 0.01). The diameters of inhibition zone of the gel against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa respectively was (15.50 ± 0.96), (15.25 ± 2.36), and (15.75 ± 1.91) mm. The half hemolysis rate (LC50) and the hemoglobin degeneration index (DI) respectively were 456 157 mg/L and 157.98%. The tea tree oil gel had no eye irritation in rabbits based on the value of LC50/DI 2 887.44. Fourteen consecutive’days administration indicated that the tea tree oil gel had no skin irritation in rabbits. The illumination score of irritative reaction to the rabbit skin was 0.125 after a single administration, while that was 0.036 after successive administration experiment. The results of high speed centrifugalization cold- resistance and heat-resistance tests showed that the preparation exhibited good stability, which needed to be kept tightly in a cool place and protected from light. Conclusion The formulation design was reasonable, while the preparation technology was simple, corresponding to the main index of the gel for topical application, with good anti-inflammatory efficacy, antibacterial effect and safety, which offered the basis for further research and development of tea tree oil.

OBJECTIVE: To investigate the protective effect of the decoction of tartary buckwheat flavonoids on focal cerebral is-chemia-reperfusion injury of rats. MEHTODS: Fifty male SD rats will be randomly divided into normal group, cerebral ischemia-reperfusion group (model group), high dose of tartary buckwheat flavonoids group, middle dose of tartary buckwheat flavonoids group and low dose of tartary buckwheat flavonoids group, each groups of 10 rats. A rat model of the right middle cerebral artery occlusion/reper-fusion(MCAO) was established by the filament method. After being operated, treatment-group rats will be administered 100,75 and 50 mg · kg-1 of the decoction of tartary buckwheat flavonoids three times a day for 7 consecutive days, after administrated for 7 d, rats in each group will undergo neurobehavioral tests. Expressions of cerebral inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) were measured by SP immunohistochemistry. The optical density value (OD) was measured by imaging analysis, and the percentage of cells with iNOS and eNOS positive expression was analyzed under light microscope. RESULTS: Compared with ischemia-reperfusion group, neurological function score increased in the decoction of tartary buckwheat flavonoids groups. Treatment groups had lower expression level of iNOS but higher expression level of eNOS than those in the model groups (P < 0.01, P < 0.05). The number of neurons of Hippocampal CA1 was increased (P < 0.05). CONCLUSION: Tartary buckwheat flavonoids can improve neurological function and decrease the expression of iNOS and increase the expression of eNOS in cerebral ischemia-reperfusion rats, which may contribute to the protection of neural function.

Objective: To study the prescription and preparation technology of tanshinone IIA for oral self-microemulsifying drug delivery system. The quality, stability, and in vitro dissolution were evaluated. Methods: The prescription and preparation technology were selected and optimized through the solubility experiment, orthogonal design, and pseudo-ternary phase diagram method, using the self-emulsifying time, appearance, particle diameter, and stability as selection indexes. The droplet morphous, particle size, drug content, stability, and in vitro dissolution were evaluated after self-microemulsification. Results: The prescription composition of tanshinone IIA self microemulsion was aethylis oleas (50%), polysorbate 80 (40%), and PEG 400 (10%), with oil phase-aqueous phase of 1:50, drug-loaded of 3.0 mg/g, and self-emulsifying time of 1 min. The acquired tanshinone IIA self-microemulsion exhibitted uniform and transparent, with the particle diameter of (51.39 ± 1.50) nm, polydispersity index of 0.211 ± 0.022, Zeta potential of (-11.35 ± 1.19) mV. The results of in vitro dissolution indicated that the accumulative dissolution in 0.1 mol/L HCl solution was able to reach 96% after 30 min. The stability result showed that tanshinone IIA self-microemulsion was affected by high temperature and illumination, indicating having to be stored at 4℃ and protected from light. Conclusion: The preparation of tanshinone IIA self-microemulsion is simple, increasing the solubility in water, making it better absorption in the stomach and intestine, corresponding to the main index of oral drug delivery system and offering the basis for further development and research about tanshinone IIA.

Hepatitis B virus (HBV) infection could lead to different clinical presentations and disease progresses, including acute and chronic hepatitis B, liver cirrhosis, hepatocellular carcinoma, and occult HBV infection, in which the interaction between virus and host plays an important role. Because HBV reverse transcriptase is lack of correction function, HBV is prone to generate mutations under the pressure of host immune response and antiviral drug treatment. Some mutations in the HBV S gene-encoding region can significantly attenuate antibody immune response against HBV and therefore affect clinical presentations and disease progression. Such mutations are termed immune escape-related mutations. In this paper, we mainly review the structure and functional characteristics of HBV S gene, the causes and forms of the immune escape-related mutation, its influence on clinical presentations and antiviral treatment response, as well as clinical detection methods.

]复发和转移是影响鼻咽癌患者预后和生存质量的主要原因。目前临床上主要使用基于解剖学的TNM分期标准，并不能 准确反映患者的预后情况，因此需要开发新的更为精准的鼻咽癌预后判断标准。通过对鼻咽癌发病机制的研究，分子标志物在 鼻咽癌预后过程的预测价值日益引起关注。本文总结了近年来在鼻咽癌增殖、生存和侵袭转移等方面的研究进展，以及在这些 研究过程中发现的相关分子标志物和它们在临床预后评估中的作用，综合分析这些分子标志物差异性表达与临床患者生存之间 的相关性，证明了这些分子标志物在评估患者预后中的潜在价值。随着对鼻咽癌发病机制研究的深入，将会发现更多有价值的 分子标志物，这为建立更为精准的鼻咽癌预后评估方法提供了可能。

Background Vascular endothelial injury is an important pathogenic step of vibration-induced hand arm vibration disease (HAVD), and long-term vibration exposure can lead to vascular endothelial cell dysfunction and cell damage. Cell ferroptosis may be one of the important mechanisms of vibration-induced vascular endothelial cell injury and HAVD. Objective To explore whether vibration can induce changes in ferroptosis-related indicators in vascular endothelial cells in vitro. Methods Human umbilical vein endothelial cells (HUVEC) were divided into four vibrationgroups and two control groups. The vibration groups were exposed to an vibration setting of 125 Hz, 6.5 m·s⁻² frequency band and for different durations: 1 d 2 h (total 1 d, 2 h per day), 1 d 4 h (total 1 d, 4 h per day), 2 d 2 h (total 2 d, 2 h per day), and 2 d 4 h (total 2 d, 4 h per day), respectively. All control groups were treated the same as the experimental groups except no vibration exposure. When the cells were 80% confluent, the control groups and the corresponding experimental groups were harvested at the same time. The effects of subgroup treatments on iron, reduced glutathione (GSH), and malondialdehyde (MDA) in HUVEC were detected with a cell ferrous colorimetric test kit, a reduced GSH colorimetric test kit, and a trace MDA test kit, respectively. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the mRNA expressions of ferroptosis-related genes acyl-CoA synthetase long chain family member 4 (ACSL4), tumor protein 53 (P53), recombinant human ferritin heavy chain (FTH1), and glutathione peroxidase 4 (GPX4). Western blot (WB) was used to detect the expression levels of ferroptosis-related proteins in HUVEC. Results Compared with the control groups, the vibration induced an increase in the iron content of HUVEC with a dose-response trend. Compared with the control groups, the reduced GSH content of HUVEC in the vibration group decreased with the increase of vibration time and frequency, and there was a dose-response trend. Compared with the control groups, the intracellular MDA content of HUVEC in the 1 d 2 h, 1 d 4 h, and 2 d 4 h vibration groups increased, and the MDA content in the 1 d 2 h and 1 d 4 h vibration group increased with time. The RT-PCR results showed that the mRNA expression levels of ACSL4 and P53 in the 1 d 4 h group increased compared with the 1 d 2 h group. Compared with the 2 d control group, the mRNA expression levels of ACSL4 in the 2 d 2 h vibration group and the 2 d 4 h vibration group increased, and the mRNA expression level of P53 in the 2 d 4 h vibration group increased. Compared with the 1 d control group, the mRNA expression levels of FTH1 and GPX4 in endothelial cells in the vibration 1 d 2 h group decreased. The WB results showed that compared with the control groups, the expression level of ferroptosis-related protein ACSL4 in endothelial cells increased in the vibration 1 d 2 h group; the expression levels of P53 in the 1 d 2 h and 2 d 4 h vibration groups increased; the expression levels of GPX4 decreased in the 1 d 4 h and 2 d 2 h vibration group, and the decrease was more obvious in the 2 d 2 h vibration group than in the 1 d 2 h vibration group; the above differences were statistically significant (P<0.05). Conclusion Vibration induces an increase in iron content, a decrease in GSH, and an increase in MDA in vascular endothelial cells in vitro, as well as mRNA and protein expressions of ferroptosis-related genes ACSL4, P53, FTH1, and GPX4.