Objective To investigate the gene expression profile in rat pancreatic ductal stem cells(PDSCs)when induced to differentiate into insulin-secreting cells(IPCs),with the goal of identifying key genes involved in this differentiation process.Methods The expanded PDSCs were categorized into a normal control(NC)group and an induced(Tre)group.PDSCs continued expansion culture in NC group,and cultured in induction medium for 28 days to facilitate the differentiation of PDSCs into IPCs in Tre group.Dithizone staining was employed to morphologically assess whether the cells exhibited a reddish-brown coloration,indicating a positive result.The immunofluorescence staining method was used to detect the expression of insulin(Ins)and PDX1 in the cells following induction.Additionally,ELISA was conducted to measure the Ins release from IPCs,thereby verifying the responsiveness of the induced cells to glucose-stimulated Ins secretion.Concurrently,cells were collected on induction days 0 and 28 for RNA sequencing(RNA-seq),and differentially expressed genes(DEGs)were analyzed and functionally annotated.The analysis revealed that regulatory factor X3(RFX3)was overexpressed in PDSCs,and the impact of RFX3 upregulation on differentiation induction was subsequently verified.Results Compared with NC group,DTZ staining was positive,PDX1 and Ins proteins were expressed,and an increased release of Ins in response to sugar stimulation was demonstrated in the Tre group.RNA-seq analysis identified 4270 DEGs,and functional enrichment analysis utilizing the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases revealed associations with Ins response,positive regulation of Ins secretion,pancreatic endocrine cell development,and overall pancreatic development.Additionally,functionally related genes such as ALDHA2,CREB5,EIF6,FOXO1,RFX3,WNT5a,OGT,GPR39,SMAD6,and TRPM2 were identified,indicating involvement in the cell cycle,TGF-β1 signaling pathway,FOXO signaling pathway,and Wnt signaling pathway in the regulation of the differentiation of pancreatic ductal stem cells(PDSCs)into insulin-producing cells(IPCs).Furthermore,the upregulation of RFX3 can inhibit the expression of TGF-β1 within 72 hours,thereby promoted the formation and release of Ins from insulin-positive cells.Conclusions Multiple genes and signaling pathways associated with pancreatic β-cell function collectively regulate the differentiation of rat PDSCs into IPCs.Notably,the upregulation of RFX3 enhances this differentiation process.

Objective To investigate the gene expression profile in rat pancreatic ductal stem cells(PDSCs)when induced to differentiate into insulin-secreting cells(IPCs),with the goal of identifying key genes involved in this differentiation process.Methods The expanded PDSCs were categorized into a normal control(NC)group and an induced(Tre)group.PDSCs continued expansion culture in NC group,and cultured in induction medium for 28 days to facilitate the differentiation of PDSCs into IPCs in Tre group.Dithizone staining was employed to morphologically assess whether the cells exhibited a reddish-brown coloration,indicating a positive result.The immunofluorescence staining method was used to detect the expression of insulin(Ins)and PDX1 in the cells following induction.Additionally,ELISA was conducted to measure the Ins release from IPCs,thereby verifying the responsiveness of the induced cells to glucose-stimulated Ins secretion.Concurrently,cells were collected on induction days 0 and 28 for RNA sequencing(RNA-seq),and differentially expressed genes(DEGs)were analyzed and functionally annotated.The analysis revealed that regulatory factor X3(RFX3)was overexpressed in PDSCs,and the impact of RFX3 upregulation on differentiation induction was subsequently verified.Results Compared with NC group,DTZ staining was positive,PDX1 and Ins proteins were expressed,and an increased release of Ins in response to sugar stimulation was demonstrated in the Tre group.RNA-seq analysis identified 4270 DEGs,and functional enrichment analysis utilizing the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases revealed associations with Ins response,positive regulation of Ins secretion,pancreatic endocrine cell development,and overall pancreatic development.Additionally,functionally related genes such as ALDHA2,CREB5,EIF6,FOXO1,RFX3,WNT5a,OGT,GPR39,SMAD6,and TRPM2 were identified,indicating involvement in the cell cycle,TGF-β1 signaling pathway,FOXO signaling pathway,and Wnt signaling pathway in the regulation of the differentiation of pancreatic ductal stem cells(PDSCs)into insulin-producing cells(IPCs).Furthermore,the upregulation of RFX3 can inhibit the expression of TGF-β1 within 72 hours,thereby promoted the formation and release of Ins from insulin-positive cells.Conclusions Multiple genes and signaling pathways associated with pancreatic β-cell function collectively regulate the differentiation of rat PDSCs into IPCs.Notably,the upregulation of RFX3 enhances this differentiation process.

Objective:To investigate the changes of hippocampal gray matter volume and expression of candidate immune related genes in a rat model of schizophrenia established by repeated administration of dizocilpine(MK-801).Methods:Thirty SPF grade Sprague-Dawley male rats at postnatal day 28 were randomly divided into MK-801 medium-dose (0.25 mg/kg) group, MK-801 high-dose(0.50 mg/kg) group and normal saline (5 mL/kg) group according to random number table method, with 10 in each group.Rats were given continuous intraperitoneal administration according to grouping once a day for 14 days.Open field test, novel object recognition test and Y-maze test were used at postnatal day 60 to detect spontaneous activity, exploration ability, anxiety level, object recognition memory ability and spatial working memory of rats, respectively.At postnatal day 67, structural magnetic resonance imaging was used to detect the changes of hippocampal gray matter volume in rat.And at postnatal day 70, qRT-PCR was used to detect the expression of candidate immune-related genes in rat hippocampus.SPSS 25.0 was used for statistical analysis, one-way ANOVA was used for comparison among multiple groups, and Tukey test was used for further pairwise comparisons.Results:(1)The behavioral results showed that there were significant differences in the total movement distance, central area activity time, novel object recognition index, and spontaneous correct alternation rate among the three groups ( F=11.15, 10.11, 13.62, 11.99, all P<0.05). The total movement distances in MK-801 medium-dose group and MK-801 high-dose group ((21.44±2.17) m, (22.87±1.96)m) were higher than that in the normal saline group ((18.70±1.88) m) (both P<0.05). The activity time of the central area in the MK-801 medium-dose group and MK-801 high-dose group((3.24±1.58) s, (2.50±1.32) s) were lower than that of the normal saline group ((6.05±2.48)s) (both P<0.01). Novel object recognition indexes in the MK-801 medium-dose group and MK-801 high-dose group((56.10±3.99)%, (54.00±6.41)%) were both lower than that in the normal saline group ((65.90±5.65)%)(both P<0.01), and the rates of spontaneous correct alternation ((54.60±7.03)%, (51.60±8.84)%) in the two groups were lower than that of the normal saline group ((68.40±8.57)%) (both P<0.01). (2) The results of structural magnetic resonance imaging showed that there were significant differences in the volume of hippocampal gray matter among the three groups ( F=9.24, P<0.001). The volumes of hippocampal gray matter in MK-801 medium-dose group and MK-801 high-dose group were lower than that in normal saline group(both P<0.001). (3)By constructing protein-protein interaction network, four candidate immune related genes were screened out: neuropeptide Y (NPY), somatostatin (SST), cholecystokinin (CCK) and tachykinin 1 (TAC1). The results showed that the mRNA expression levels of NPY, SST and CCK in the hippocampus of the three groups were significantly different ( F=11.41, 10.43, 5.85, all P<0.05), but there was no statistical difference in the TAC1 mRNA expression level ( F=0.08, P>0.05). The mRNA levels of NPY, SST and CCK in the hippocampus of rats in the MK-801 high-dose group were lower than those in the normal saline group (all P<0.05). Conclusion:Both medium dose and high dose MK-801 administration can reduce the volume of hippocampal gray matter in schizophrenia model rats, but they have different effects on the expression of hippocampal immune related genes, of which high dose administration has a greater effect.

OBJECTIVE To analyze the risk factors of substandard drug blood concentration of meropenem in patients with hospital acquired pneumonia （HAP）. METHODS Totally 130 HAP patients who were admitted to the intensive care unit of Suzhou Hospital Affiliated to Nanjing Medical University from January 2020 to June 2021 and received steady -state blood concentration test of meropenem were selected as the study subjects . The patient ’s age ，sex，body mass and other medical history were recorded . The steady-state blood trough concentration of meropenem was determined and its target was determined . Univariate and multivariate Logistic regression analysis were used to screen the risk factors for the substandard steady -state blood trough concentration of meropenem. The receiver operating characteristic （ROC）curve was drawn to screen the warning value of the risk factors and evaluate the predictive value of the risk factors . RESULTS The steady -state blood trough concentrations of 85 cases were ≥2 mg/L， and those of 45 cases were ＜2 mg/L. Multivariate Logistic regression analysis showed that age ，negative balance and brain injury were independent risk factors for the substandard steady-state blood trough concentration of meropenem （P＜ 0.05）.ROC curve showed that when the patient was 58 years old，the area under the ROC curve was the largest （0.744）， the sensitivity was 0.882，the specificity was 0.556，and the Youden index was 0.438；when the negative balance was 520.5 mL/24 h，the area under the ROC curve reached the maximum （0.827），the sensitivity was 0.722，the specificity was 0.905，and th e Youden index was 0.628. The creatinine clearance rate in the brain injury group was significantly higher than that in the non -brain injury group ，and the steady -state blood trough concentration of meropenem in the brain injury group was significantly lower than that in the non -brain injury group （P＜0.001）. CONCLUSIONS When the HAP patient ’s age is less than 58 years old ，the brain injury and the negative balance is more than 520.5 mL/24 h，the risk of substandard steady -state blood trough concentration of meropenem will increase .

Objective To investigate the affective and somatic symptoms of patients with somatoform disorders (SFD) and their related factors to their social support and alexithymia. Methods Seventy-one patients with SFD and 75 healthy controls (HC) were recruited. Health Questionnaire?9 (PHQ?9), General Anxiety Scale (GAD?7), Patient Health Questionnaire 15?Item Somatic Symptom Severity Scale (PHQ?15), Toronto Alexithymia Scale (TAS?20) and Social Support Rating Scale (SSRS) were evaluated for all subjects. Results (1) The total scores of all the PHQ?9, GAD?7 and PHQ?15 in patients group were significantly higher than that in HC (t=46.67, 41.24, 49.64, P<0.01). (2) Except the objective social support, subject social support and social support utilization in SFD group were lower than those in HC (36.97±6.74 vs. 40.54±5.47, 19.34±3.17 vs. 23.53±3.62, 6.41±3.13 vs. 8.36±2.43; t=7.46, 4.28, 3.52; P<0.01). There were significant differences in TAS?20 between the two groups (t=8.08, 8.52, 5.54, 7.35, P<0.01). (3) By the correlation analysis in patient group, the sum score of the affective and somatic symptom (PHQ?9+GAD?7+PHQ?15) was significantly correlated to the subjective social support, social support utilization, social support total score, TAS?Ⅰ, TAS?Ⅱ, TAS?Ⅲand TAS?20 total score (r=-0.372,-0.359,-0.426, 0.368, 0.327, 0.306, 0.364, P<0.01). And the numbers of non?mental health clinic visits (χ1), social support utilization (χ2), adverse reactions(χ3), TAS?Ⅱ (χ4) and TAS?Ⅰ (χ5) contributed to the multiple regression equation for the sum of PHQ?9, GAD?7 and PHQ?15 scores. Conclusion The alexithymia and incapability of perceiving and utilizing social support, numbers of visits in non?mental health clinic and the medication adverse reactions that subjectively perceived in the patients with SFD may be the key factors that could impact their affective and somatic symptoms.

Objective To investigate the affective and somatic symptoms of patients with somatoform disorders (SFD) and their related factors to their social support and alexithymia. Methods Seventy-one patients with SFD and 75 healthy controls (HC) were recruited. Health Questionnaire?9 (PHQ?9), General Anxiety Scale (GAD?7), Patient Health Questionnaire 15?Item Somatic Symptom Severity Scale (PHQ?15), Toronto Alexithymia Scale (TAS?20) and Social Support Rating Scale (SSRS) were evaluated for all subjects. Results (1) The total scores of all the PHQ?9, GAD?7 and PHQ?15 in patients group were significantly higher than that in HC (t=46.67, 41.24, 49.64, P<0.01). (2) Except the objective social support, subject social support and social support utilization in SFD group were lower than those in HC (36.97±6.74 vs. 40.54±5.47, 19.34±3.17 vs. 23.53±3.62, 6.41±3.13 vs. 8.36±2.43; t=7.46, 4.28, 3.52; P<0.01). There were significant differences in TAS?20 between the two groups (t=8.08, 8.52, 5.54, 7.35, P<0.01). (3) By the correlation analysis in patient group, the sum score of the affective and somatic symptom (PHQ?9+GAD?7+PHQ?15) was significantly correlated to the subjective social support, social support utilization, social support total score, TAS?Ⅰ, TAS?Ⅱ, TAS?Ⅲand TAS?20 total score (r=-0.372,-0.359,-0.426, 0.368, 0.327, 0.306, 0.364, P<0.01). And the numbers of non?mental health clinic visits (χ1), social support utilization (χ2), adverse reactions(χ3), TAS?Ⅱ (χ4) and TAS?Ⅰ (χ5) contributed to the multiple regression equation for the sum of PHQ?9, GAD?7 and PHQ?15 scores. Conclusion The alexithymia and incapability of perceiving and utilizing social support, numbers of visits in non?mental health clinic and the medication adverse reactions that subjectively perceived in the patients with SFD may be the key factors that could impact their affective and somatic symptoms.

Objective To explore effects of dizocilpine (MK-801) preconditioning on excitatory amino acids and inflammatory response in rats induced by cardiac arrest-cardiopulmonary resuscitation (CACPR).Methods 18 male Sprague Dawley (SD) rats were randomly divided into three groups:control group,CA group and CA + MK-801 group.To establish rat models of CA-CPR and keep samples of serum and specimens of brain tissues for following detection.The injury of neurons was observed by HE staining and expression of N-methyl-D-aspartic acid receptor (NMDAR) in brain tissues was detected by Western blot.The concentrations of interleukin 1 beta (IL-1 β) and tumor necrosis factor (TNF)-α in serum were detected by enzyme linked immunosorbent assay (ELISA).Results Neurons in CA group were disorganized,cells shrank,nuclei pyknosis,and cytoplasmic eosinophilia,accompanied by inflammatory cell infiltration.Preconditioning with MK-801 reduced the pathological damage of neuron and degree of macrophage infiltration.The relative expression of NMDAR protein in CA group were significantly higher than that in control group (907.9 ±24.9 vs 321.6 ± 18.4,P ＜0.001).Preconditioning with MK-801 significantly decreased the expression of NMDAR in CA + MK-801 group compared with that in CA group (512.4 ± 21.1 vs 907.9 ± 24.9).The CA group showed significantly increased concentrations of IL-1 β and TNF-α than that in control group (P ＜ 0.001),and this effect was abolished by preconditioning with MK-801.CA rats treated with MK-801 showed higher concentrations of IL-1 β and TNF-α than the control group.Conclusions Cardiac arrest causes pathological injury of neurons,up-regulates expression of NMDAR and aggravates inflammatory response.These results induce the apoptosis of nerve cells.Blocking glutamate receptor with MK-801 can inhibit expression of NMDAR,decrease level of cytokines,down-regulate inflammatory reaction degree therefore to protect the brain.

Objective To evaluate the miRNA change between hypothermia circulatory arrest at different temperature and its impact on intestinal protection.Methods Sixteen piglets were randomly(n =4) divided into four groups:deep hypothermia circulatory arrest (DHCA,18℃) group,moderate hypothermia circulatory arrest(MHCA,24℃) group,cardiopulmonary bypass(CPB) group and sham operation(SO) group.They were subjected to 80 min hypothermia circulatory arrest,305 min CPB or thoracotomy,respectively.Pick-and-mix custom miRNA real-time PCR panels were utilized to detect intestinal samples.miRNA expression between DHCA and MHCA were compared directly(DHCA vs.MHCA) and indirectly(DHCA/SO vs.MHCA/SO,DHCA/CPB vs.MHCA/CPB).Results Exposure to DHCA caused less intestinal miRNA dysregulation than MHCA.Besides,seven miRNAs(miR-122,miR-145-5p,miR-421-5p,miR-99a,miR-365-5p,miR-31 and miR-192)were differentially expressed between the two hypothermia circulatory arrest groups.Conclusion Better intestinal miRNA protection was provided by DHCA than MHCA.Intestinal miRNA were differentially expressed between hypothermia circulatory arrest at different temperature.

Objective To study the pharmacokinetic features of ferulaic acid, senkyunolide A and ligustilide in Buyang Huanwu associated prescriptions (Buyang HuanwuDecoction andNaojianTablets).MethodsHPLC-DAD was applied for simultaneous determination plasma concentration of three ingredients with jugular venous cannula rats after intragastric administration ofBuyang Huanwu associated prescriptions. The pharmacolinetic parameters of each ingredient was calculated by DAS2.0, and then the total quantum statistical moment (TQSM) standard similarity was used to measure the overall pharmacokinetics behaviors.Results There were great differences in the three ingredients after the administration of two prescriptions, while the total quantum statistical parameters were very closely. The TQSM pharmacokinetic parameters of the three components inBuyang HuanwuDecoction andNaojian Tablets showed that AUC, MRT, VRT were 240.6 and 133.0, 3.192 min and 3.259 min, 21.59 min2and 19.75 min2, respectively.The similarity was up to 0.977 8.Conclusion The metabolic processes in vivo ofBuyang Huanwu Decoction andNaojianTablets have similarities. The efficacy of Chinese herbal compounds mostly depends on the multi-components overall contributions.

Objective To explore the expression of TLR4/NF-κB pathway in cerebral injury resulting from DHCA ( deep hypothermia circulatory arrest ) as well as the effect of SACP ( selective antegrade cerebral perfusion). Methods Twelve pigs were randomly assigned to DHCA group (n = 6) or SACP group (n = 6) at 18 ℃ for 80 min. IL-6 was assayed by ELISA. Apoptosis and NF-κB proteins were detected by fluorescence TUNEL and Western blot, respectively. The level of TLR4 was determined through qRT-PCR and Western blot. Results Serum IL-6 level of SACP group was significantly lower at the end of circulation arrest and experiment and apoptotic index and NF-κB protein were apparently lower in SACP group (P < 0.05). The level of TLR4 protein and mRNA from SACP group decreased significantly (P < 0.05). Conclusions TLR4/NF-κB pathway plays a critical role in pathogenesis of DHCA cerebral injury and attenuating TLR4/NF-κB cytokines probably contributes to neuroprotection of SACP. TLR4/NF-κB pathway may be a novel target for DHCA.