Objective To investigate the effect of acidic serine protease ASPNJ on the expression of heat shock protein HSP90, 60 and 27 in human chronic myeloid leukemia K562 cells, in order to reveal the related mechanism of anti leukemic effects of ASPNJ. Methods K562 leukemia cell lines were cultured in vitro and treated with ASPNJ alone or in combination with chemotherapeutic agents. Western blot and RT-PCR were used to detect the changes of HSP90, 60 and 27 gene expressions in levels of total protein and membrane protein, as well as in mRNA levels. Results ASPNJ showed different effects on the expression of HSPs in total protein and membrane protein levels and had some modified effect on HSPs in total protein or membrane protein levels. Effects of ASPNJon expression of HSPs mRNA were not apparent, but HSPs mRNA were apparently lower in the ASPNJ and doxorubicin combination group than that in the ASPNJ alone or doxorubicin alone groups. Conclusion The mechanism of ASPNJ on the inhibitory effect of leukemia cells proliferation and the promoting effects on chemotherapeutic drugs may involve some complicated correlations with the effect of ASPNJ on the expression of HSPs and the modification of HSPs proteins.

Objective To investigate the effects of fentanyl citrate on pain in aged rats with hip fracture and its underlying mechanisms.Methods Thirty aged rats with hip fracture were divided in-to model group(n=10),fentanyl citrate group(n=10),and fentanyl citrate+Sar-SP group(n=10).Additionally,healthy rats from the same period were included as normal group(n=10).Rats in the fentanyl citrate group received intravenous tail injection of 10 pg/kg fentanyl citrate,while those in the fentanyl citrate+Sar-SP group underwent intrathecal injection of 10 μL Sar-SP for fenta-nyl citrate intervention.Rats in the normal and model groups received intravenous tail injection of an equal volume of saline.Pain threshold,weight-bearing capacity,temperature,and dorsoventral thick-ness of the hind paws were measured before treatment(0h)and at 1,6,24,and 168 h post-treat-ment.Enzyme-linked immunosorbent assay(ELISA)was used to detect levels of tumor necrosis fac-tor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β),and glutamic acid(Glu)in the spinal cord tissue of rats.Immunohistochemical staining was employed to observe the positive expression rates of ionized calcium-binding adapter molecule 1(Iba-1),glial fibrillary acidic protein(GFAP),and substance P(SP)in the spinal cord tissue.Real-time fluorescent quantitative polymerase chain re-action(qRT-PCR)was conducted to detect the relative expression levels of SP and tachykinin re-ceptor 1(TACR1)mRNA in the hippocampus tissue of rats.Western blot analysis was performed to assess the relative expression levels of vimentin(VIM),nuclear receptor corepressor 1(NOCR),ciliary neurotrophic factor receptor(CNTFR),and epidermal growth factor receptor(EGFR)pro-teins in the spinal cord tissue.Results Compared with the normal group,rats in the model group exhibited decreased pain threshold and weight-bearing capacity,increased hind paw temperature and dorsoventral thickness,as well as elevated levels of TNF-α,IL-6,IL-1 β,and Glu,increased posi-tive expression rates of Iba-1 and GFAP,heightened positive expression rate and relative mRNA ex-pression levels of SP and TACR1,and augmented relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 0 h before treatment and at 1,6,24,and 168 h post-treatment(P＜0.05).Compared with the model group,rats in the fentanyl citrate group showed increased pain threshold and weight-bearing capacity,decreased hind paw temperature and dorsoventral thickness,as well as reduced levels of TNF-α,IL-6,IL-1β,and Glu,decreased posi-tive expression rates of Iba-1 and GFAP,lowered positive expression rate and relative mRNA expres-sion levels of SP and TACR1,and decreased relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 1,6,24,and 168 h post-treatment(P＜0.05).Compared with the fentanyl citrate group,rats in the fentanyl citrate+Sar-SP group demonstrated decreased pain threshold and weight-bearing capacity,increased hind paw temperature and dorso-ventral thickness,elevated levels of TNF-α,IL-6,IL-1 β,and Glu,increased positive expression rates of Iba-1 and GFAP,heightened positive expression rate and relative mRNA expression levels of SP and TACR1,and elevated relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 1,6,24,and 168 h post-treatment(P＜0.05).Conclusion The im-provement in pain in aged rats with hip fracture by fentanyl citrate may be associated with the reduc-tion of SP expression in the spinal cord that inhibits glial cell activation and exhibits anti-inflammato-ry effects.

Objective To investigate the effects of fentanyl citrate on pain in aged rats with hip fracture and its underlying mechanisms.Methods Thirty aged rats with hip fracture were divided in-to model group(n=10),fentanyl citrate group(n=10),and fentanyl citrate+Sar-SP group(n=10).Additionally,healthy rats from the same period were included as normal group(n=10).Rats in the fentanyl citrate group received intravenous tail injection of 10 pg/kg fentanyl citrate,while those in the fentanyl citrate+Sar-SP group underwent intrathecal injection of 10 μL Sar-SP for fenta-nyl citrate intervention.Rats in the normal and model groups received intravenous tail injection of an equal volume of saline.Pain threshold,weight-bearing capacity,temperature,and dorsoventral thick-ness of the hind paws were measured before treatment(0h)and at 1,6,24,and 168 h post-treat-ment.Enzyme-linked immunosorbent assay(ELISA)was used to detect levels of tumor necrosis fac-tor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β),and glutamic acid(Glu)in the spinal cord tissue of rats.Immunohistochemical staining was employed to observe the positive expression rates of ionized calcium-binding adapter molecule 1(Iba-1),glial fibrillary acidic protein(GFAP),and substance P(SP)in the spinal cord tissue.Real-time fluorescent quantitative polymerase chain re-action(qRT-PCR)was conducted to detect the relative expression levels of SP and tachykinin re-ceptor 1(TACR1)mRNA in the hippocampus tissue of rats.Western blot analysis was performed to assess the relative expression levels of vimentin(VIM),nuclear receptor corepressor 1(NOCR),ciliary neurotrophic factor receptor(CNTFR),and epidermal growth factor receptor(EGFR)pro-teins in the spinal cord tissue.Results Compared with the normal group,rats in the model group exhibited decreased pain threshold and weight-bearing capacity,increased hind paw temperature and dorsoventral thickness,as well as elevated levels of TNF-α,IL-6,IL-1 β,and Glu,increased posi-tive expression rates of Iba-1 and GFAP,heightened positive expression rate and relative mRNA ex-pression levels of SP and TACR1,and augmented relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 0 h before treatment and at 1,6,24,and 168 h post-treatment(P＜0.05).Compared with the model group,rats in the fentanyl citrate group showed increased pain threshold and weight-bearing capacity,decreased hind paw temperature and dorsoventral thickness,as well as reduced levels of TNF-α,IL-6,IL-1β,and Glu,decreased posi-tive expression rates of Iba-1 and GFAP,lowered positive expression rate and relative mRNA expres-sion levels of SP and TACR1,and decreased relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 1,6,24,and 168 h post-treatment(P＜0.05).Compared with the fentanyl citrate group,rats in the fentanyl citrate+Sar-SP group demonstrated decreased pain threshold and weight-bearing capacity,increased hind paw temperature and dorso-ventral thickness,elevated levels of TNF-α,IL-6,IL-1 β,and Glu,increased positive expression rates of Iba-1 and GFAP,heightened positive expression rate and relative mRNA expression levels of SP and TACR1,and elevated relative protein expression levels of VIM,NOCR,CNTFR,and EGFR in the spinal cord tissue at 1,6,24,and 168 h post-treatment(P＜0.05).Conclusion The im-provement in pain in aged rats with hip fracture by fentanyl citrate may be associated with the reduc-tion of SP expression in the spinal cord that inhibits glial cell activation and exhibits anti-inflammato-ry effects.

Visual hallucination(VH)is a common symptom of schizophrenia,the underlying mechanism has not been fully elucidated.It has been found that the dysfunction of dopamine(DA)system,the overactivation of α-amino-3-hydroxy-5-methyl-4-isoxazole-propionate(AMPA)receptor in glutamate system and the dysfunction of γ-aminobutyric acid(GABA)ergic neurons can induce VH in patients with schizophrenia.In addition,abnormalities in brain structural and functional networks and visual networks are also closely related to the occurrence of VH.The purpose of this paper is to review the neurochemistry and nerve injury mechanism of VH in schizophrenic patients to deeply understand the characteristics of VH,and make more accurate judgment in the early diagnosis,condition evaluation and treatment plan of schizophrenic patients.

Objective:To study the genotyping characteristics of Brucella strains isolated from Lianyungang City (non-brucellosis epidemic area) of Jiangsu Province. Methods:Preliminary identification of 13 suspected strains of Brucella isolated from blood culture in Clinical Microbiology Laboratory of the First People's Hospital of Lianyungang City in 2018 was conducted; at the same time, the specific gene bcsp31 and insertion sequence IS-711 of Brucella were detected by quantitative real-time PCR (Real-time PCR), and the identification results were rechecked and typed. Multiple locus variable-number tandem repeat analysis (MLVA) was applied for genotyping, and the sequencing results were edited by Mega 4.0 software. Results:All the 13 strains were identified as Brucella by preliminary identification. Real-time PCR confirmed that all the 13 strains were Brucella melitensis. The results of MLVA showed that 13 strains of Brucella melitensis were divided into 12 genotypes and clustered in the "middle Mediterranean cluster". Among 13 strains of Brucella melitensis, 3 strains were biovar 1, 2 strains were biovar 2 and 8 strains were biovar 3. Conclusion:All the Brucella strains isolated from Lianyungang City are Brucella melitensis and the MLVA cluster is in the "middle Mediterranean cluster".

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.

Geriatric depression in the elderly is becoming one of the most common psychiatric disorders affecting older people's mental and physical health.However, there is currently no systematic review on animal models for geriatric depression.Therefore, this paper analyzes and summarizes the animal models commonly used in geriatric depression studies and the application of antidepressants in geriatric depression models based on relevant national and international literature of recent years, aiming to provide insights on research approaches and considerations on study methods for geriatric depression.

Objective:To compare the expression levels of candidate genes before and after Shuganjieyu capsule treatment, to analyze their correlation with depression symptoms and cognitive function, and to find and clarify the biomarkers related to the efficacy of Shuganjieyu capsule.Methods:Among 27 patients with mild to moderate depression (MMD), 24 items Hamilton depression rating scale (HAMD-24) was used to assess the severity of depression, Chinese revised Wechsler adult intelligence scale(WAIS-RC) and Chinese revised Wechsler memory scale(WMS) were used to assess cognitive function, and qRT-PCR was used to detect the expression levels of candidate genes in peripheral blood of patients with depression before and after treatment with Shuganjieyu capsule.SPSS 25.0 software was used for statistical analysis, paired t-test, non-parametric test, Spearman correlation analysis and receiver operating characteristic curve were used for data statistics. Results:The symptoms of MMD patients were relieved after Shuganjieyu capsule treatment(HAMD scores: baseline 14.00(9.75, 18.25), 8-week 4.00(2.00, 7.25), Z=-4.462, P<0.01), and the verbal intelligence quotient(VIQ) of WMS was puomoved (VIQ scores: baseline (123.00±10.24), 8-week (128.00±6.77), t=4.372, P<0.01). The level of gene expression brain derived neurotrophic factor(BDNF) (baseline 1.68(0.92, 2.63), 8-week 2.30(1.47, 4.34), Z=-2.781, P=0.005), glial cell derived neurotrophic factor(GDNF) (baseline 0.74(0.31, 1.15), 8-week 0.97(0.50, 1.71), Z=-2.159, P=0.031), 5-hydroxytryptamine receptor 2A(HTR2A) (baseline 0.60(0.39, 1.60), 8-week 0.98(0.44, 2.29), Z=-1.994, P=0.046) and glutamate ionotropic receptor AMPA type subunit 1(GRIA1) (baseline 1.19(0.66, 2.40), 8-week 1.76(0.86, 4.13), Z=-2.756, P=0.006) was up-regulated after treatment.The change rate of BDNF expression were correlated with the score of HAMD-24 ( r=-0.35, P=0.038) and performance intelligence quotient of WMS ( r=0.40, P=0.022). Conclusions:BDNF may be used as a therapeutic marker of Shuganjieyu capsule in the treatment of clinical symptoms and cognitive function of MMD patients, which is used to evaluate the efficacy of antidepressants.