Humans ; PPAR gamma/metabolism* ; Multiple Sclerosis ; Transcription Factors/metabolism* ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha

OBJECTIVE To investigate the factors influencing the changes in purchasing quantity in the procurement varieties of the first batch of volume-based drug centralized procurement （hereinafter referred to as centralized procurement）. METHODS Using 25 procurement varieties of the “4+7” policy as research objects， the changes in purchasing quantity of procurement varieties were analyzed before and after the implementation of the “4+7” pilot， renewal and expansion policies. The influential factors were determined from the three levels of drugs， medical institutions and regions； and the multiple linear regression model was used to analyze the influential factors for the changes in the purchasing quantity of procurement varieties. RESULTS Before and after the implementation of the “4+7” pilot， renewal and expansion policies， the purchasing quantity increased by 52.1， －0.2， 85.8 ten thousand DDDs on average， compared with base period. During pilot， renewal and expansion period， DDDc decrease in procurement varieties was positively correlated with the increase in purchasing quantity （P＜0.01）. During the pilot and renewal period， the number of absolutely alternative varieties was positively correlated with the increase in purchasing quantity （P＜0.1）. During the pilot and expansion period， the number of alternative varieties to a certain extent was negatively correlated with the increase in purchasing quantity （P＜0.05）. During the renewal period， the increment of purchasing quantity in tertiary hospitals was smaller than that of primary medical institutions （P＜0.05）. CONCLUSIONS There is a relationship between the decline of DDDc and the changes in the purchasing quantity， that is， the more the drug price dropped， the more the purchasing quantity increased. The number of alternative varieties for centralized procurement will affect the changes in their purchasing quantity， but it is not always stable. With the implementation of the policy， the volume of primary medical institutions gradually exceeds that of tertiary institutions， indicating that the consumption of centralized purchased varieties is transferred to the primary medical institutions， and centralized procurement has promoted the implementation of the hierarchical diagnosis and treatment system.

Objective:To investigate the regulatory effect of transient receptor potential cation channel subfamily C member 3 (TRPC3) on the retina in oxygen-induced retinopathy (OIR) mice and biological behavior of human retinal vascular endothelial cells (HREC).Methods:A total of 32 healthy SPF grade 7-day-old C57BL/6 mice were selected and randomly divided into a control group and an OIR group by the random number table method, with 16 mice in each group.The control group received no special treatment, and the OIR model was established in the OIR group.On postnatal day 17 (PN17), the success of the model establishment was verified by immunofluorescence staining of the retinal patch.The in vitro cultured HREC were divided into a normal control group, a transfection reagent group, and a si-TRPC3 group.The normal control group received no special treatment, while the transfection reagent group and the si-TRPC3 group were transfected with transfection reagent or transfection reagent + si-TRPC3.The relative expression of TRPC3 mRNA was detected by real-time quantitative fluorescence PCR.The relative expressions of TRPC3, transcription factor NF-E2 related factor (Nrf2), and superoxide dismutase (SOD) proteins were determined by Western blot.HREC were further divided into a normal control group, a vascular endothelial growth factor (VEGF) group, a si-TRPC3 group, and a Pyr3 (TRPC3 channel inhibitor) group, which were cultured in complete medium, medium containing 20 ng/ml VEGF recombinant protein, medium containing 20 ng/ml VEGF recombinant protein (si-TRPC3 transfection for 72 hours), and medium containing 20 ng/ml VEGF recombinant protein+ 1 μmol/L Pyr3 for 48 hours, respectively.The proliferation ability of HREC was detected using cell counting kit 8 (CCK-8). The horizontal and vertical migration ability of cells were detected by cell scratch assay and transwell assay, respectively.This study followed the 3R principles of animal welfare and was approved by the Ethics Committee of Hebei Eye Hospital (No.2023LW04). Results:Pathological neovascular clusters with strong fluorescent staining appeared in the retina of OIR mice on PN17.The relative expressions of TRPC3 mRNA and protein in the retina of OIR mice were 2.057±0.244 and 1.517±0.290, respectively, significantly higher than 0.983±0.033 and 0.874±0.052 of control group ( t=6.165, 3.094; both at P<0.05). The relative expression levels of TRPC3 mRNA and protein were significantly lower, and the relative expression levels of Nrf2 and SOD proteins were higher in the si-TRPC3 group than in the normal control and transfection reagent groups, and the differences were statistically significant (all at P<0.05). The CCK-8 experiment results showed that the cell absorbance value was higher in the VEGF group than in the normal control group, and lower in the si-TRPC3 and Pyr3 groups than in the VEGF group, with statistically significant differences (all at P<0.05). The results of the cell scratch experiment showed that the lateral migration rate of VEGF group cells was higher than that of normal control group, while the lateral migration rate of si-TRPC3 group and Pyr3 group cells was lower than that of VEGF group, and the differences were statistically significant (all at P<0.05). The transwell experiment results showed that the number of stained cells in the VEGF group was higher than that in the normal control group, and the number of stained cells in the si-TRPC3 group and Pyr3 group was lower than that in the VEGF group, with statistically significant differences (all at P<0.05). Conclusions:Hypoxia induces increased TRPC3 expression in OIR mouse retina, and downregulation of TRPC3 inhibits HREC proliferation and migration.The mechanism is related to the activation of the Nrf2-related oxidative stress pathway.

BACKGROUND:Icariin,with antiinflammatory,antioxygenatory and immunoregulatory effects,can be a potential drug for preventing and treating acute liver injury. OBJECTIVE:To investigate the protective effect and possible mechanism of icariin in mice with acute liver injury induced by carbon tetrachloride. METHODS:Thirty-two Kunming mice were equally and randomly divided into the following groups:normal,model,low-dose icariin and high-dose icariin groups.The low-and high-dose icariin groups were continuously gavaged with icariin(100 and 200 mg/kg,respectively)once a day for 7 continuous days.The normal group and model group were injected with physiological saline(10 mL/kg)at the same time point.After the last administration,all the groups except for the normal group were injected with carbon tetrachloride to induce acute liver injury.The mice were killed 24 hours later,and the liver index was detected.Serum levels of alanine aminotransferase and aspartate aminotransferase were detected by automated biochemical analysis.Tumor necrosis factor α and interleukin 6 levels in serum were detected using ELISA.The levels of superoxide dismutase,glutathione peroxidase and malondialdehyde in liver tissue were detected through a reagent kit.The histopathology changes of the liver were observed by hematoxylin-eosin staining.TUNEL method was used to detect the apoptosis in hepatocytes.Western blot was performed to detect the expression levels of glucose-regulated protein 78 kDa,endoplasmic reticulum stress-related protein(C/-EBP homologous protein),mixed lineage kinase domain-like protein and Caspase-3 in liver tissue. RESULTS AND CONCLUSION:Compared with the normal group,the liver index and serum levels of alanine aminotransferase,aspartate aminotransferase,tumor necrosis factor α and interleukin 6 were increased in the model group(P＜0.05).Compared with the model group,the above indexes were decreased in the low-dose and high-dose icariin groups(P＜0.05).Compared with the normal group,the activities of superoxide dismutase and glutathione peroxidase in the liver tissue of mice were decreased in the model group(P＜0.05)and the level of malondialdehyde was increased(P＜0.05).Compared with the model group,the activities of superoxide dismutase and glutathione peroxidase were increased in the low-and high-dose icariin groups(P＜0.05)and the level of malondialdehyde was decreased(P＜0.05).Hematoxylin-eosin and TUNEL staining showed that mice in the model group had severe structural destruction of liver tissue,extensive necrosis of hepatocytes and high apoptotic rate of hepatocytes,while the structural destruction of liver tissue and the area of necrosis of hepatocytes in the low-and high-dose icariin groups were significantly milder than those in the model group,and the apoptotic rate of hepatocytes was lower than that in the model group(P＜0.05).Western blot assay showed that the protein expression of glucose-regulated protein 78 kDa,C/-EBP homologous protein,mixed lineage kinase domain-like protein and Caspase-3 in liver tissue of mice in the model group was increased compared with that in the normal group(P＜0.05),while the expression levels of these proteins in liver tissue of mice were significantly reduced after low-and high-dose icariin intervention(P＜0.05).To conclude,icariin can produce a protective effect against carbon tetrachloride-induced acute liver injury,and its mechanism may be related to the regulation of endoplasmic reticulum stress and reduction of programmed necrosis.

BACKGROUND:The pituitary gland is an important endocrine organ in the body.Certain diseases can cause damage to the pituitary gland,such as pituitary adenoma and abnormal hormone secretion.Pituitary stem cells,due to their self-renewal and multi-directional differentiation potential,are expected to become a new therapeutic approach for repairing damaged pituitary glands. OBJECTIVE:To isolate and culture pituitary stem cells using the suspension cell ball culture method and identify their proliferation and differentiation ability. METHODS:Pituitary stem cells were isolated and cultured from the pituitary gland of newborn New Zealand white rabbits using the suspension cell ball culture method,and their morphological characteristics were observed.Immunofluorescence cytochemistry was used to detect the expression of pituitary stem cell markers SOX2 and Nestin.EdU labeling method was utilized to detect the proliferative ability of pituitary stem cells.After adherent and induced differentiation,the hormone levels in the culture medium were detected by ELISA. RESULTS AND CONCLUSION:Pituitary stem cell spheres could be successfully isolated by the suspension cell ball culture method,with strong proliferative ability.Positive expression of stem cell-specific markers SOX2 and Nestin was found in the cultured cells.After induction and differentiation,adrenocorticotropic hormone,thyroid hormone,growth hormone,luteinizing hormone,follicle-stimulating hormone,and prolactin levels significantly increased in the medium(P<0.001),with strong differentiation ability.

Objective:To investigate the impact of programmed death receptor-1(PD-1)and its ligand(PD-L1)on the differentiation of decidual macrophages(DMs)and embryonic development.Methods:20 patients with re-current spontaneous abortion(RSA group)and 20 patients with normal pregnancy induced artificial abortion(NP group)were selected from Sichuan Provincial People's Hospital between March 2021 and January 2024.Villous and decidual tissues were obtained from both groups.Real-time fluorescent quantitative polymerase chain reac-tion(qPCR)and Western blot techniques were used to detect the expression of PD-L1 mRNA and protein in vil-lous tissues.Flow cytometry was used to analyze the types of DMs and their PD-1 expression.Human monocytic leukemia cells(THP-1)were differentiated under different conditions,with a control group and a PD-1 blocked group established,and different reagents were added into those two groups to induce differentiation,then the pro-portion of DMs types was measured in both groups.Results:PD-L1 protein and mRNA were expressed in the vil-lous tissue of both groups of patients but the expression of PD-L1 protein and mRNA was reduced in the RSA group compared to the NP group(P<0.05).The decidual tissues of both groups contained M1 and M2 macro-phages.Compared with the NP group,the ratio of M2 to M1 DMs in the RSA group was lower(P<0.05),indica-ting a polarization towards the M1 type in DMs.There was no statistically significant difference in PD-1 expression between the two groups(P>0.05).Human THP-1 cells can be stimulated to differentiate into M1 and M2 macro-phages.Afterblocking PD-1,theratioof M2 to M1 macrophages decreased(P<0.05),indicating a polarization towards the M1 type in macrophages.Conclusions:In patients with RSA,the expression of PD-L1 in villous tis-sues is decreased,and DMs polarize towards the M1 type.Blocking the PD-1/PD-L1 pathway can further promote the polarization of macrophages towards the M1 type.The influence of PD-1/PD-L1 pathway on macrophage po-larization may provide new insights into the pathogenesis of RSA.

Objective:To investigate the impact of programmed death receptor-1(PD-1)and its ligand(PD-L1)on the differentiation of decidual macrophages(DMs)and embryonic development.Methods:20 patients with re-current spontaneous abortion(RSA group)and 20 patients with normal pregnancy induced artificial abortion(NP group)were selected from Sichuan Provincial People's Hospital between March 2021 and January 2024.Villous and decidual tissues were obtained from both groups.Real-time fluorescent quantitative polymerase chain reac-tion(qPCR)and Western blot techniques were used to detect the expression of PD-L1 mRNA and protein in vil-lous tissues.Flow cytometry was used to analyze the types of DMs and their PD-1 expression.Human monocytic leukemia cells(THP-1)were differentiated under different conditions,with a control group and a PD-1 blocked group established,and different reagents were added into those two groups to induce differentiation,then the pro-portion of DMs types was measured in both groups.Results:PD-L1 protein and mRNA were expressed in the vil-lous tissue of both groups of patients but the expression of PD-L1 protein and mRNA was reduced in the RSA group compared to the NP group(P<0.05).The decidual tissues of both groups contained M1 and M2 macro-phages.Compared with the NP group,the ratio of M2 to M1 DMs in the RSA group was lower(P<0.05),indica-ting a polarization towards the M1 type in DMs.There was no statistically significant difference in PD-1 expression between the two groups(P>0.05).Human THP-1 cells can be stimulated to differentiate into M1 and M2 macro-phages.Afterblocking PD-1,theratioof M2 to M1 macrophages decreased(P<0.05),indicating a polarization towards the M1 type in macrophages.Conclusions:In patients with RSA,the expression of PD-L1 in villous tis-sues is decreased,and DMs polarize towards the M1 type.Blocking the PD-1/PD-L1 pathway can further promote the polarization of macrophages towards the M1 type.The influence of PD-1/PD-L1 pathway on macrophage po-larization may provide new insights into the pathogenesis of RSA.

Objective:To investigate the impact of programmed death receptor-1(PD-1)and its ligand(PD-L1)on the differentiation of decidual macrophages(DMs)and embryonic development.Methods:20 patients with re-current spontaneous abortion(RSA group)and 20 patients with normal pregnancy induced artificial abortion(NP group)were selected from Sichuan Provincial People's Hospital between March 2021 and January 2024.Villous and decidual tissues were obtained from both groups.Real-time fluorescent quantitative polymerase chain reac-tion(qPCR)and Western blot techniques were used to detect the expression of PD-L1 mRNA and protein in vil-lous tissues.Flow cytometry was used to analyze the types of DMs and their PD-1 expression.Human monocytic leukemia cells(THP-1)were differentiated under different conditions,with a control group and a PD-1 blocked group established,and different reagents were added into those two groups to induce differentiation,then the pro-portion of DMs types was measured in both groups.Results:PD-L1 protein and mRNA were expressed in the vil-lous tissue of both groups of patients but the expression of PD-L1 protein and mRNA was reduced in the RSA group compared to the NP group(P<0.05).The decidual tissues of both groups contained M1 and M2 macro-phages.Compared with the NP group,the ratio of M2 to M1 DMs in the RSA group was lower(P<0.05),indica-ting a polarization towards the M1 type in DMs.There was no statistically significant difference in PD-1 expression between the two groups(P>0.05).Human THP-1 cells can be stimulated to differentiate into M1 and M2 macro-phages.Afterblocking PD-1,theratioof M2 to M1 macrophages decreased(P<0.05),indicating a polarization towards the M1 type in macrophages.Conclusions:In patients with RSA,the expression of PD-L1 in villous tis-sues is decreased,and DMs polarize towards the M1 type.Blocking the PD-1/PD-L1 pathway can further promote the polarization of macrophages towards the M1 type.The influence of PD-1/PD-L1 pathway on macrophage po-larization may provide new insights into the pathogenesis of RSA.

Objective:To investigate the impact of programmed death receptor-1(PD-1)and its ligand(PD-L1)on the differentiation of decidual macrophages(DMs)and embryonic development.Methods:20 patients with re-current spontaneous abortion(RSA group)and 20 patients with normal pregnancy induced artificial abortion(NP group)were selected from Sichuan Provincial People's Hospital between March 2021 and January 2024.Villous and decidual tissues were obtained from both groups.Real-time fluorescent quantitative polymerase chain reac-tion(qPCR)and Western blot techniques were used to detect the expression of PD-L1 mRNA and protein in vil-lous tissues.Flow cytometry was used to analyze the types of DMs and their PD-1 expression.Human monocytic leukemia cells(THP-1)were differentiated under different conditions,with a control group and a PD-1 blocked group established,and different reagents were added into those two groups to induce differentiation,then the pro-portion of DMs types was measured in both groups.Results:PD-L1 protein and mRNA were expressed in the vil-lous tissue of both groups of patients but the expression of PD-L1 protein and mRNA was reduced in the RSA group compared to the NP group(P<0.05).The decidual tissues of both groups contained M1 and M2 macro-phages.Compared with the NP group,the ratio of M2 to M1 DMs in the RSA group was lower(P<0.05),indica-ting a polarization towards the M1 type in DMs.There was no statistically significant difference in PD-1 expression between the two groups(P>0.05).Human THP-1 cells can be stimulated to differentiate into M1 and M2 macro-phages.Afterblocking PD-1,theratioof M2 to M1 macrophages decreased(P<0.05),indicating a polarization towards the M1 type in macrophages.Conclusions:In patients with RSA,the expression of PD-L1 in villous tis-sues is decreased,and DMs polarize towards the M1 type.Blocking the PD-1/PD-L1 pathway can further promote the polarization of macrophages towards the M1 type.The influence of PD-1/PD-L1 pathway on macrophage po-larization may provide new insights into the pathogenesis of RSA.

Objective To explore the protective mechanism of honey-processed Hedysari Radix in regulating intestinal mucosal injury in rats with spleen qi deficiency.Methods The three-factor composite modeling method of bitter cold diarrhea,overwork and hunger and satiety disorder was used to construct a spleen qi deficiency model rats.After the model was successfully made,they were randomly divided into model group,honey-processed Hedysari Radix group and probiotic group,with 15 animals in each group.Another 15 normal rats were taken as the blank group.The honey-processed Hedysari Radix group was given 12.6 g·kg-1 water decoction of honey-processed Hedysari Radix by gavage,the probiotics group was given Bifidobacterium Lactobacillus triple viable tablets suspension at a dose of 0.625 g·kg-1,and the blank group and the model group were given the same dose of distilled water.The rats in the four groups were administered once a day for 15 days.Enzyme-linked immunosorbent assay was used to detect diamine oxidase(DAO)in serum,D-lactic acid(D-LA),secretory immunoglobulin A factor,and Western blotting was used to detect the expression levels of AMP-activated protein kinase(AMPK),zonula occludens-1(ZO-1)and occludin in colon tissues.Results The serum levels of DAO in the blank group,model group,honey-processed Hedysari Radix group and probiotic group were(138.93±9.78),(187.95±12.90),(147.21±6.92)and(166.47±3.37)pg·mL-1;the contents of D-LA were(892.23±49.17),(1 099.84±137.64),(956.56±86.04)and(989.61±51.75)μg·L-1;the contents of SIgA in colon tissues were(14.04±1.42),(11.47±2.39),(11.84±1.49)and(12.93±1.65)μg·mL-1;the relative expression levels of ZO-1 protein in colon tissues were 1.18±0.11,0.42±0.04,0.77±0.05 and 0.95±0.07;the relative expression levels of occludin protein were 1.35±0.31,0.61±0.17,1.19±0.19 and 0.88±0.13;the relative expression levels of AMPK protein were 0.91±0.02,0.35±0.09,0.74±0.08 and 0.59±0.11.Compared with the model group,there were significant differences in the serum content of DAO and D-LA,SIgA content in colon,and the content of ZO-1,occludin and AMPK protein in the honey-processed Hedysari Radix group(P＜0.01,P＜0.05).Conclusion Honey-processed Hedysari Radix can enhance the protective effect on the intestinal mucosa of rats with spleen qi deficiency by regulating the expression of related inflammatory cytokines,intestinal mucosal upper cell enzymes and tight junction proteins in rats with spleen qi deficiency.