Objective To investigate the amlodipine or hydrochlorothiazide and valsartan combination thera-py in elderly hypertensive patients with blood pressure variability ( BPV ) and nitric oxide ( NO ) , endothelin role. Methods 552 elderly patients with hypertension were divided into the observation group and control group,276 cases in each group,the observation group was given amlodipine valsartan for treatment,while the control group was received hydrochlorothiazide combined with valsartan for treatment,the two group were treated for three months,the content of BPV,NO and plasma endothelin were analyzed.Results After treatment,the 24hSBPV,day SBPV,24hDBPV,day DBPV of the observation group significantly decreased compared with pre-treatment, the difference was statistically significant(t=20.09,15.97,9.31,9.41,all P<0.05),patients in the control group only 24hSBPV and day SBPV [(9.12 ±2.57)%,(8.43 ±1.97)%]were significantly lower than before treatment(t=12.31,9.53,all P<0.05);The day SBPV,24hDBPV,day DBPV[(7.21 ±1.34)%,(12.31 ±2.54)%,(10.59 ±2.73)%]of the ob-servation group after treatment were significantly lower than those of the control group(t=10.52,12.34,8.35,all P<0.05);The NO[ (66.12 ±23.57)%,(68.29 ±21.52)%]and endothelin levels[(34.43 ±7.97)%,(34.21 ± 7.34)%]of the two groups after treatment were significantly lower than those of before treatment(tobservation grou P=31.39,11.79,tcontrol grouP=28.31,9.35,all P<0.05).Conclusion The amlodipine or valsartan hydrochlo-rothiazide used clinically can effectively control blood pressure, significantly increase NO levels, decrease plasma endothelin levels,which is worth to be used in the clinical.

The pollen grains of 24 cultivarieties of Primus mimic of Rosaceae, which were collected from Shanghai, Hangzhou and Changxing. were observed by light microscope and scanning electron microscope. The pollen morphology of P.mume was first described as polytypism which could be distinguish into two classes and five types. Not only the pollen grains with 3-colporate. which were the primary form, but also the pollen grains with 4-colporate existed in many cultivarieties of P- tnume. The constitute of pollen types and sterility rate in each cultivarieties were also analysed. The result provides scientific reference for pollen identification, breeding and palynology.

Objective To investigate the roles of glutamate signaling pathway in melanin transfer.MethodsEpidermal melanocytes and keratinocytes were isolated from human foreskin tissue followed by purification and primary culture.Immunofluorescence microscopy was conducted to observe the intracellular distribution of N-methy-D-aspartate receptor 1 (NMDAR1) and NMDAR2A in melanocytes.Some melanocytes were classified into 4 groups to be pretreated with MK801 (the NMDAR antagonist dizocilpine maleate) at 100μmol/L for 5 minutes followed by treatment with NMDA(an NMDAR agonist) at 100 μmol/L (MK801-pretreated group 1),pretreated with MK801 at 100 μmol/L for 1 hour followed by treatment with NMDA at 100μmol/L (MK801-pretreated group 2),treated with MK801 at 100 μmol/L for 5 minutes (MK801 group),treated with NMDA at 100 μmol/L for 5 minutes (NMDA group),respectively,then,confocal microscopy was performed to measure the intracellular calcium (Ca2+) concentration of the melanocytes.The distribution of β-tubulin was visualized by confocal microscopy in melanocytes treated with MK801 at 100 μmol/L for 24 hours.Some melanocytes and keratinocytes were cocultured with or without MK801 at 100 μmol/L for 24 or 48 hours,then,scaning microscopy was carried out to observe the junction structure between melanocytes and keratinocytes,and alkali method coupled with spectrophotometric analysis to determine melanin content in keratinocytes.Results The intracellular calcium concentration of melanocytes was decreased by MK-801,but increased by NMDA at 100 μmol/L,and the increase was blocked by the pretreatment with MK-801 for 5 minutes or 1 hour.After incubation with MK-801 at 100 μmol/L for 24 hours,a more intense staining for β-tubulin was observed around the nuclei of melanocytes.There was a significant reduction in the number of filopodia on the surface of and between melanocytes and keratinocytes after treatment with MK-801 at 100 μmol/L for 48 hours.Also,the content of melanin(represented as the absorbance value at 375 nm) transferred from melanocytes into keratinocytes was statistically reduced in coculture system treated with MK-801 at 100 μmol/L compared with that without treatment (0.158 ± 0.003 vs.2.203 ± 0.006,t =6.323,P ＜ 0.01 ).Conclusions The glutamate signaling pathway exerts a regulatory effect on intracellular calcium concentration of distribution of β-tubulin in,filopodia formation of melanocytes and melanin transfer between melanocytes and keratinocytes.

Objective To investigate the action mechanism of glutamate-mediated signaling pathway in malignant melanoma. Methods WM451LU melanoma cells in log phase were classified into 6 groups, negative control group treated with PBS (100 μl), MK801 group treated with the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 (100 μmol/L), CPCCOEt group treated with non-competitive metabotropic glutamate receptor 1 (mGluR1) antagonist CPCCOEt, MAP2 group transfected with adenovirus vector containing microtubule associated protein 2a (Ad-MAP2a), MK801 + MAP2 group treated with MK801 of 100 μmol/L and transfected with Ad-MAP2a, CPCCOEt + MAP2 group treated with CPCCOEt of 10 μmol/L and transfected with Ad-MAP2a. Western blot was performed to detect the expression of an ionotropic glutamate receptor, i.e., N-methyl-D-aspartate receptor type 2A (NMDAR2A) in WM451LU cells transfected with Ad-MAP2a. Scratch motility assay and cell invasion assay were conducted in vitro to detect the changes in migration and invasion ability of WM451LU cells after treated with Ad-MAP2a, MK-801, CPCCOEt alone or in combination. In vivo study was carried out to compare the inhibitory effect of the above treatments on melanoma. Results Western blot revealed a decrease in the expression of NMDAR2A in WM451LU cells after transfected with Ad-MAP2a. The scratch motility assay showed that the number of migrating cells per high power field was 117.04 ± 2.76 in MAP2 group,107.64 ± 6.50 in MK801 group,97.36 ± 4.79 in CPCCOEt group, 43.28 ± 3.02 in MK801 + MAP2 group,30.76 ± 3.97 in CPCCOEt + MAP2 group,significantly different from that in the negative control group (152.3 ± 5.75,all P ＜ 0.01 ). Cell invasion assay demonstrated that the average number of invading cells per high power field in the negative control was significantly higher than that in MAP2 group, MK801 group, CPCCOEt group, MK801+MAP2 group and CPCCOEt + MAP2 group (170.43 ±8.72 vs. 98.26 ± 3.84, 97.22 ± 5.54, 112.23 ± 7.21, 42.89 ± 5.06, 58.25 ± 6.68, P ＜ 0.05, 0.05, 0.05, 0.01and 0.01, respectively).A significant decrease was observed in the average volume of experimental melanoma in mice of MAP2 group, MK801 group, MK801 + MAP2 group, CPCCOEt group and CPCCOEt + MAP2 group compared with the negative control group (224.02 ± 46.19 mm3, 160.33 ± 33.91 mm3, 91.49 ± 21.48 mm3,202.30 ± 52.37 mm3, 111.13 ± 69.81 mm3 vs. 342.70 ± 60.92 mm3, all P ＜ 0.01 ). Conclusions To block the glutamate signaling pathway in vitro can inhibit the invasion and migration of melanoma cells, and to block the pathway in vivo can inhibit the growth of malignant melanoma and alter the morphology of melanoma cells.

OBJECTIVE To e xplore the research hots pots and development frontiers of ursolic acid in recent 20 years，and to provide reference for researchers in this field. METHODS Research literatures related to ursolic acid in Web of Science from Jan. ， 1，2002 to Dec. 31，2021 were collected ，and visualization analysis was performed on countries or regions ，research institutions ， authors，journals and keywords involved in the literatures using CiteSpace software ，to obtain the spatial and temporal distribution of ursolic acid and research frontiers. The research status and development frontier of ursolic acid were further analyzed by analyzing keywords co-occurrence ，keyword emergence ，keyword clustering ，etc. RESULTS & CONCLUSIONS Totally 3 528 valid papers were included in this study ，and the top three countries were China ，India and the United States. Analysis of publishing institutions showed that Chinese Academy of Sciences ，Univisity of Karachi and China Medical University were the top 3 research institutions in the list of publication amount. Analysis of published journals showed that Molecules （127 articles），Journal of Ethnopharmacology（90 articles），Journal of Agricultural and Food Chemistry （75 articles）had high number of literatures on ursolic acid. The analysis of keyword analysis showed that pharmacological effects ，such as antitumor activity of ursolic acid ， antioxidant activity ，antibacterial activity and anti-inflammatory activity ，are always the focus of the research ；the mechanism ursolic acid induced apoptosis ，oxidative stress and autophagy ，the research on ursolic acid signaling pathway ，drug delivery of ursolic acid nanoparticles were the research direction in the future.

OBJECTIVE：To explo re the potential molecular mechanism of ursolic acid in the treatment of osteoporosis （OP）. METHODS：TCMSP，PubMed database and UniProt database were used to screen potential targets of monomer compound ursolic acid. OP related target genes were searched with GeneCards database. The common target genes of component-disease were obtained by Venny 2.1 online mapping tool. The protein-protein interaction （PPI）network of component-disease common target genes was constructed by using STRING database ，and topological analysis was carried out ；the core target genes ，whose degree value was greater than the average degree value ，were screened. GO functional annotation and KEGG pathway enrichment analysis of component-disease common target genes were carried out by DAVID database. AutoDock Vina 1.1.2 software was used for molecular docking ，using protein encoded by the core target gene as receptor and ursolic acid as ligand. RESULTS ：A total of 55 ursolic acid related target genes and 4 273 OP related target genes were excavated ，with a total of 44 common target genes. PPI network with above common target genes included 44 nodes and 513 edges，with an average node degree of 23.3. There were 24 core target genes ，including VEGFA，TP53，IL6，CASP3. There were 340 GO functional items were enriched （corrected P＜ 0.05），including 263 biological processes （negative regulation of apoptosis ，etc.），25 molecular functions （protein binding ，etc.） and 52 cell components （cytosol，etc.）. There were 90 KEGG signaling pathways （corrected P＜0.05），such as tumor pathway ， hepatitis B pathway ，TNF signaling pathway ，viral carcinogenesis and phosphatidylinositol 3 kinase/protein kinase B （PI3K-Akt） signaling pathway. The binding energy between ursolic acid and 6 proteins encoded by core target genes such as TP53 was lower than －5 kcal/mol，which had strong binding activity. CONCLUSIONS ：The therapeutic effect of ursolic acid on OP may be achieved by regulating VEGFA，TP53，IL6，CASP3，JUN and other core target genes and acting on multiple key pathways such as cancer pathway ， hepatitis B and TNF signaling