Objective To observe the effect of different fixative solutions on cancer cell morphology and membrane permeability. Methods Human pancreatic acinar epithelial carcinona(HPAC) cells of human pancreatic cancer and HeLa cells of human cervical cancer were fixed with 4 fixation solutions: freshly prepared 0.25% paraformaldehyde solution; Freshly prepared 4% paraformaldehyde solution; 75% ethanol solution; 90% ethanol solution. The fixation time is 30 minutes. PBS solution and complete medium were used as the controls. Cell morphology of each group was observed under optical microscope. Changes in cell membrane permeability were observed by fluorescence staining with 7-aminoactinomycin (7-AAD), which is not cell membrane permeable in intact cells but permeable in damaged cells. Hoechst33342 was used for staining both intact and damaged cells. Results The cells in the complete medium group were similar to unfixed cells in morphology, and the fluorescence staining of 7-AAD was the weakest. The cells in the complete medium group have typical cell morphology and low 7-AAD permeability. The 0.25% paraformaldehyde solution group had similar cell morphology to the complete medium group, and the 7-AAD fluorescence staining was weak. The morphology of cells in the 4% paraformaldehyde solution group was typical, but the fluorescence staining of 7-AAD was strong. The cells in the 90% ethanol solution group showed swelling, with a larger volume than the unfixed cells and a stronger fluorescence staining of 7-AAD. The cell swelling in 75% ethanol solution group was not as obvious as that in 90% ethanol solution group, and the fluorescence staining of 7-AAD was strong. The cells in PBS group were round, and the fluorescence staining of 7-AAD was strong. Conclusion 0.25% paraformaldehyde solution can not only fix tumor cells, but also maintain the integrity of cell membrane.

 Objective To analyse the analgesic effect and possible mechanism of panax notoginseng saponin (PNS) on mouse models of chronic inflammatory pain caused by complete Freund’s adjuvant (CFA).   Methods A total of 48 male C57BL/6J mice were divided randomly into four groups: normal saline control group (Ctrl), CFA group (CFA), CFA + PNS group (CFA+PNS), CFA + dexamethasone (DEX) group (CFA+DEX). Von Frey filaments were used to detect mechanical pain in mice. Immunohistochemistry was used to detect the number and morphological changes of glial fibrillary acidic protein (GFAP) positive astrocytes. Western blotting was used to detect the expressions of GFAP, nucleotide-binding and oligomerization domain(NOD)-like receptor thermal protein domain associated protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, interleukin (IL)-1β, and IL-18 in mice’s spinal cord segments in each group.   Results Compared with the Ctrl group, mice in the CFA group showed a significant decrease in mechanical pain thresholds at day 1, day 3, day 5, day 7, and day 14. Additionally, there was a significant decrease in NLRP3, ASC, Caspase-1, IL-1β and IL-18 in the spinal cord of the mice. PNS intervention could relieve mechanical pain and down-regulate the expressions of NLRP3, ASC, Caspase-1, IL-1β and IL-18 in the spinal cord of mice, with no significant difference compared with the CFA+DEX group. CFA group mice had significantly more GFAP positive cells in their posterior horns than Ctrl group mice, as measured by immunohistochemistry; PNS intervention decreased the number of GFAP positive cells in the posterior horn of the spinal cord in model mice；DEX had no effect on the number of GFAP positive cells in the dorsal horn of spinal cord. According to Western blotting results, GFAP expression in the spinal cord of the CFA group was significantly more than that of the Ctrl group; PNS intervention significantly reduced GFAP expression in the spinal cord of CFA group mice；DEX had no effect on the expression of GFAP in the posterior horn of spinal cord.   Conclusion PNS has a good alleviating effect on inflammatory pain, and its mechanism may be related to inhibition of astrocyte activation and NLRP3 inflammasome activation. 

Objective  To observe the expression and localization of group Ⅰ metabotropic glutamate receptors (mGluR1/5) in rat superior cervical ganglion (SCG) and the effect of chronic intermittent hypoxia (CIH) on mGluR1/5 protein level.   Methods  Twelve male SD rats were randomly divided into control group（Ctrl）and CIH group（CIH）, 6 rats in each group. After 6 weeks of modeling, the effect of CIH on mGluR1/5 protein level was detected by Western blotting, the expression and distribution of mGluR1/5 in SCG were detected by immunohistochemistry and double-immunofluorescent staining.     Results  mGluR1/5 was expressed in rat SCG. mGluR1 was distributed in neurons and small intensely fluorescent (SIF) cells, but not in satellite glial cells (SGCs), nerve fibers and blood vessels, whereas mGluR5 was mainly distributed in nerve fibers and a little in neurons, but not in SGCs, SIF cells and blood vessels. CIH increased the protein levels of mGluR1/5 (P<0.01) in rat SCG.   Conclusion  Both mGluR1 and mGluR5 are expressed in the rat SCG, but their distribution are different, and the increased protein levels of both may be involved in CIH-induced hypertension.

Objective To measure and compare the lateral posterior tibial slope (LPTS), medial posterior tibial slope (MPTS) and tibial torsion angle (TTA) between the patients of recurrent patellar dislocation and the heathy people, and to analyze the correlation between LPTS, MPTS and TTA and the risk factors of recurrent patellar dislocation.   Methods  A total of 33 patients (44 knees) with recurrent patellar dislocation in our hospital from July 2019 to June 2021 were selected and listed as the study group. Twenty-three subjects (46 knees) who were suspected iliac vascular and lower limb vascular diseases during the same period were selected and listed as the control group. All the enrolled researchers had full-length CT scans date of the lower limbs. Three-dimensional models were reconstructed using Mimics 21.0 software and then imported into 3-matic software. The LPTS, MPTS and TTA were measured and compared between the two groups.      Results In the study group, the LPTS, MPTS and TTA were （7.69±1.42）°,（10.06±1.71）°,（36.42±8.13）°, respectively, while the control group, the LPTS, MPTS and TTA were（8.42±1.65）°, （10.44±0.86）°, (25.77±3.90)°, respectively. There were no significant differences in the LPTS, MPTS and TTA between different genders and sides both in the study group and the control group (P>0.05). Compared with the control group, the LPTS in the study group was smaller, and the difference was statistically significant (P<0.05). There was no statistically significant difference between the study group and the control group in the MPTS (P>0.05). Compared with the control group, the TTA in the study group was higher, and the difference was statistically significant (P<0.05). Compared with the control group, the LPTS and MPTS in the study group were significant asymmetry, and the difference was statistically significant (P<0.05).  Conclusion The lateral posterior tibial slope of patients with recurrent patellar dislocation is significantly smaller than that in the healthy people, while there is no significant difference in the medial posterior tibial slope; The tibial torsion angle of patients with recurrent patellar dislocation is significantly larger than in the healthy people; The lateral posterior tibial slope and tibial torsion angle have certain correlation with recurrent patellar dislocation, which can conduct the diagnosis of recurrent patellar dislocation.

Objective To improve the fixation method  of the transmission electron microscope for better morphological preservation of mitochondria and lipid droplets in mouse brown adipose tissue.   Methods The fixation method  for mouse brown adipose tissue was optimized, mainly including an increased concentration of paraformaldehyde from 2% to 4% in the pre-fixative, employment of transcardial perfusion followed by immersion fixation in pre-fixation, and using imidazole-buffered osmium tetroxide as the post-fixative. The ultrastructures of brown adipocytes prepared by the improved method  were observed and compared with those of a known standard protocol (3 mice in each group). The improved method  was further validated in the quantitative analysis of mitochondrial cristae density and lipid droplets.   Results The mitochondrial cristae and membrane structure of other organelles of brown adipocytes were better preserved using the optimized method  compared with those of the standard method. Lipid droplets were presented as round structures with high electron density instead of vacuolated appearances. Using this method, we observed that the density of mitochondrial cristae and the content of lipid droplets increased in brown adipocytes after cold adaptation.   Conclusion The optimized method can better preserve the ultrastructure of organelles in brown adipocytes, especially mitochondria and lipid droplets, and may be applicable for studying the ultrastructures remodeling of brown adipose tissue under different physiological or pathological conditions.

Osteoporosis (OP), a systemic bone metabolism disease, mainly manifested in the decrease of bone mass, the increase of bone fragility and the microstructure degeneration of the bone. Along with the in-depth research of the pathogenesis of osteoporosis, the imbalance differentiation of bone marrow mesenchymal stem cell (BMSCs) (Osteogenic differentiation decrease and adipogenic differentiation increase) is the main reason that causes osteoporosis. In this paper, we summarize the signal pathways of osteogenic differentiation and adipogenic differentiation of BMSCs. Better understand these signal pathways is conducive to elucidate and treat osteoporosis.