ObjectiveBy comparing the composition and content of volatile components in raw products， wine-washed products and wine-fried products of Ligusticum sinense cv. Chaxiong rhizome（LSCR）， to investigate the influence of wine processing on the volatile components of LSCR， in order to provide a basis for the development of quality standards for LSCR and its processed products. MethodsElectronic nose was used to identify the odors of LSCR， wine-washed and wine-fried LSCR， and their volatile components were detected by headspace gas chromatography-mass spectrometry（HS-GC-MS）， and the relative mass fractions of these components were determined by peak area normalization method. Principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were performed on the obtained sample data by SIMCA 14.1 software， and the differential components of LSCR， wine-washed and wine-fried LSCR were screened according to the variable importance in the projection（VIP） value>1. Pearson correlation analysis was used to explore the relationship between volatile differential flavor components and electronic nose sensors. ResultsElectronic nose detection results showed that there were significant differences in the odors of LSCR， wine-washed and wine-fried LSCR， mainly reflected in the sensors S2， S4， S5， S6， S11， S12， S13. And a total of 62 compounds were identified from LSCR and its wine-processed products， among which 46， 50 and 51 compounds were identified from LSCR， wine-fried and wine-washed LSCR， respectively. There were 21 differential components between the raw products and wine-fried products， of which 10 components were increased and 11 were decreased after processing. There were 20 differential components between the raw products and wine-washed products， of which 11 constituents increased and 9 decreased after processing. There were 17 differential components between the wine-wash products and wine-fried products. Compared with the wine-washed products， the contents of 13 components in the wine-fried products increased， and the contents of 4 components decreased. The increasing trend of the content of phthalides in the wine-washed products was more obvious than that in the wine-fried products， but the content of total volatile components was higher in the wine-fried products than the wine-washed products. Correlation analysis showed that there were different degrees of correlation between the 7 differential sensors of electronic nose and 24 differential volatile components， mainly phthalides and olefins. ConclusionThe odor and the content of volatile components in LSCR changed obviously after wine processing， and n-butylphthalide， Z-butylidenephthalide and E-ligustilide can be used as the candidate differential markers of volatile components in LSCR before and after wine processing.

Objective:To investigate the effects of PRND downregulation on the proliferation, migration, and invasion capabilities of human renal carcinoma cells.Methods:Clinical and transcriptomic data from renal carcinoma patients were analyzed using the TCGA database, with bioinformatics methods employed for differential gene expression analysis and survival analysis [including overall survival (OS) and disease-free survival (DFS)]. Postoperative pathological specimens from 50 renal carcinoma patients admitted to Affiliated Cancer Hospital of Zhengzhou University between January 2022 and January 2023 were collected for immunohistochemical staining to assess PRND expression in renal carcinoma tissues. Two distinct small interfering RNAs (siRNAs) were used to downregulate PRND expression in renal carcinoma cell lines ACHN and 769P. Quantitative real-time polymerase chain reaction (qPCR) and Western blotting were performed to validate the knockdown efficiency of PRND at the mRNA and protein levels. The proliferation, migration, and invasion capabilities of ACHN and 769P cells were evaluated using the Cell Counting Kit-8 (CCK-8), cell migration assay, and invasion assay, which was compared between the negative control group (NC) and the two PRND knockdown groups (si1 and si2). Western blotting was used to measure the expression levels of MMP-9, E-cadherin, C-myc, Vimentin, β-catenin, and PD-L1 proteins in ACHN and 769P cell lines.Results:TCGA database analysis revealed that PRND expression was significantly higher in renal carcinoma tissues compared with adjacent normal tissues (1.172 vs. 0.383, P<0.01). Survival analysis indicated that high PRND expression was significantly negatively correlated with both OS ( P<0.01) and DFS ( P<0.01). CCK-8 assay results showed no statistically significant differences in cell viability between the experimental and control groups at 6 hours (ACHN-si1: 1.238±0.659, ACHN-si2: 1.437±0.359, ACHN-NC: 3.234±2.165, P>0.05). However, significant differences were observed at 24 hours (ACHN-si1: 5.608±0.716, ACHN-si2: 7.088±0.308, ACHN-NC: 9.764±1.088, P<0.01), 48 hours (ACHN-si1: 40.422±1.419, ACHN-si2: 41.238±2.623, ACHN-NC: 65.823±4.337, P<0.01), and 72 hours (ACHN-si1: 53.667±4.565, ACHN-si2: 54.533±2.572, ACHN-NC: 78.800±0.265, P<0.01). Similar trends were observed in 769P cells (6 hours: P>0.05; 24 hours: P<0.05; 48 and 72 hours: P<0.01). Cell migration assays demonstrated significantly reduced migration in the experimental groups (ACHN-si1: 31±10, ACHN-si2: 62±19, ACHN-NC: 175±45, P<0.01; 769P-si1: 79±16, 769P-si2: 62±14, 769P-NC: 236±77, P<0.05). Invasion assays also showed significant suppression in the experimental groups (ACHN-si1: 13±9, ACHN-si2: 15±8, ACHN-NC: 54±12, P<0.01; 769P-si1: 17±13, 769P-si2: 19±17, 769P-NC: 91±29, P<0.01). Western blotting revealed that C-myc, β-catenin, MMP-9, Vimentin, and PD-L1 protein levels were lower in the experimental groups, while E-cadherin expression was higher compared to the control groups. Conclusions:PRND is significantly overexpressed in renal carcinoma tissues and closely associated with poor patient prognosis. Downregulation of PRND markedly inhibits the proliferation, migration, and invasion of renal carcinoma cells, potentially through modulation of epithelial-mesenchymal transition (EMT)-related proteins and key molecules involved in tumor metastasis.

Objective:To investigate the effects of PRND downregulation on the proliferation, migration, and invasion capabilities of human renal carcinoma cells.Methods:Clinical and transcriptomic data from renal carcinoma patients were analyzed using the TCGA database, with bioinformatics methods employed for differential gene expression analysis and survival analysis [including overall survival (OS) and disease-free survival (DFS)]. Postoperative pathological specimens from 50 renal carcinoma patients admitted to Affiliated Cancer Hospital of Zhengzhou University between January 2022 and January 2023 were collected for immunohistochemical staining to assess PRND expression in renal carcinoma tissues. Two distinct small interfering RNAs (siRNAs) were used to downregulate PRND expression in renal carcinoma cell lines ACHN and 769P. Quantitative real-time polymerase chain reaction (qPCR) and Western blotting were performed to validate the knockdown efficiency of PRND at the mRNA and protein levels. The proliferation, migration, and invasion capabilities of ACHN and 769P cells were evaluated using the Cell Counting Kit-8 (CCK-8), cell migration assay, and invasion assay, which was compared between the negative control group (NC) and the two PRND knockdown groups (si1 and si2). Western blotting was used to measure the expression levels of MMP-9, E-cadherin, C-myc, Vimentin, β-catenin, and PD-L1 proteins in ACHN and 769P cell lines.Results:TCGA database analysis revealed that PRND expression was significantly higher in renal carcinoma tissues compared with adjacent normal tissues (1.172 vs. 0.383, P<0.01). Survival analysis indicated that high PRND expression was significantly negatively correlated with both OS ( P<0.01) and DFS ( P<0.01). CCK-8 assay results showed no statistically significant differences in cell viability between the experimental and control groups at 6 hours (ACHN-si1: 1.238±0.659, ACHN-si2: 1.437±0.359, ACHN-NC: 3.234±2.165, P>0.05). However, significant differences were observed at 24 hours (ACHN-si1: 5.608±0.716, ACHN-si2: 7.088±0.308, ACHN-NC: 9.764±1.088, P<0.01), 48 hours (ACHN-si1: 40.422±1.419, ACHN-si2: 41.238±2.623, ACHN-NC: 65.823±4.337, P<0.01), and 72 hours (ACHN-si1: 53.667±4.565, ACHN-si2: 54.533±2.572, ACHN-NC: 78.800±0.265, P<0.01). Similar trends were observed in 769P cells (6 hours: P>0.05; 24 hours: P<0.05; 48 and 72 hours: P<0.01). Cell migration assays demonstrated significantly reduced migration in the experimental groups (ACHN-si1: 31±10, ACHN-si2: 62±19, ACHN-NC: 175±45, P<0.01; 769P-si1: 79±16, 769P-si2: 62±14, 769P-NC: 236±77, P<0.05). Invasion assays also showed significant suppression in the experimental groups (ACHN-si1: 13±9, ACHN-si2: 15±8, ACHN-NC: 54±12, P<0.01; 769P-si1: 17±13, 769P-si2: 19±17, 769P-NC: 91±29, P<0.01). Western blotting revealed that C-myc, β-catenin, MMP-9, Vimentin, and PD-L1 protein levels were lower in the experimental groups, while E-cadherin expression was higher compared to the control groups. Conclusions:PRND is significantly overexpressed in renal carcinoma tissues and closely associated with poor patient prognosis. Downregulation of PRND markedly inhibits the proliferation, migration, and invasion of renal carcinoma cells, potentially through modulation of epithelial-mesenchymal transition (EMT)-related proteins and key molecules involved in tumor metastasis.

Objective To explore the feasibility of different histochemical staining in threedimensional reconstruction of peripheral nerves.Methods The samples were collected from fresh adult common tibial nerve,which were serially horizontally sliced with the thickness of 6 (m.All slices were randomly divided into four groups with different staining such as Karnovsky-Roots staining,acetylcholinesterase staining,acetyltransferase staining or carbonic anhydrase staining.The stain characteristics in various stain phrases were observed under optic-microscope.Results The character of nerve fibers could be distinguished clearly by Karnovsky-Roots,acetylcholinesterase,acetyltransferase and acetyltransferase staining.It took 50 to 52 hours to finish the staining by using KaruovskyRoots staining.Carbonic anhydrase staining was the simplest method.Conclusion Among the four staining methods,Karnovsky-Roots staining is suitable for three-dimensional reconstruction of peripheral nerves.

Objective To explore the feasibility of different histochemical staining in threedimensional reconstruction of peripheral nerves.Methods The samples were collected from fresh adult common tibial nerve,which were serially horizontally sliced with the thickness of 6 (m.All slices were randomly divided into four groups with different staining such as Karnovsky-Roots staining,acetylcholinesterase staining,acetyltransferase staining or carbonic anhydrase staining.The stain characteristics in various stain phrases were observed under optic-microscope.Results The character of nerve fibers could be distinguished clearly by Karnovsky-Roots,acetylcholinesterase,acetyltransferase and acetyltransferase staining.It took 50 to 52 hours to finish the staining by using KaruovskyRoots staining.Carbonic anhydrase staining was the simplest method.Conclusion Among the four staining methods,Karnovsky-Roots staining is suitable for three-dimensional reconstruction of peripheral nerves.