Objective To investigate the mechanism by which suppressor of cytokine signaling 3(SOCS3)regulates microglial inflammation through nuclear factor-kappaB(NF-κB),providing novel mechanistic insights into microglial involvement in Parkinson's disease(PD)pathogenesis.Methods ① Ten male C57BL/6 mice(12 weeks old,weighing 20～25 g)were subjected to intraperitoneal injection of 15 mg/kg MPTP to establish a PD model.Rotarod test was used to assess motor function.Western blotting was employed to detect the protein expression of tyrosine hydroxylase(TH)and ionized calcium-binding adapter molecule 1(IBA-1)in the substantia nigra.RT-qPCR was utilized to measure the mRNA level of SOCS3 in the substantia nigra.Immunohistochemistry was performed to assess NF-κB p65 subunit expression.The expression of SOCS3,NF-κB and p-NF-κB was measured with Western blotting.② Microglial cell line BV2 was stimulated with 1 000 ng/mL lipopolysaccharide(LPS)for 6 h to establish an inflammatory model.Subsequently,SOCS3 was knocked down.NF-κB inhibitor BAY 11-7082 was used to treat the cells.RT-qPCR and Western blotting were used to measure the expression of SOCS3 at mRNA and protein levels.Western blotting was also applied to detect the expression of NF-κB and p-NF-κB,and ELISA was conducted to measure TNF-α and IL-1β levels in the culture supernatant.Immunofluorescence assay was carried out to localize NF-κB(nuclear vs cytoplasmic).③ A co-culture system of BV2 microglia and N2a neuroblastoma cells was established to investigate the regulatory effects of microglia on neuronal cells.MTT assay and TUNEL staining were used respectively to determine cell viability and apoptosis of N2a cells.Results ① Compared to the control mice,the PD mouse model exhibited reduced rotarod fall latency,down-regulation in TH and SOCS3(P<0.01),up-regulation in IBA-1 and increased p-NF-κB/NF-κB ratio(P<0.01).② In BV2 cells,LPS stimulation increased TNF-α,IL-1β,and p-NF-κB/NF-κB ratio(P<0.01),while down-regulated SOCS3 expression(P<0.01).SOCS3 knockdown in LPS-stimulated BV2 cells further increased the p-NF-κB/NF-κB ratio(P<0.01),increased nuclear localization of NF-κB,and elevated TNF-α and IL-1β levels(P<0.01).BAY 11-7082 treatment in these SOCS3-knockdown,LPS-stimulated cells resulted in reduced p-NF-κB/NF-κB ratio,TNF-α,and IL-1β(P<0.01),and decreased NF-κB nuclear distribution.③ LPS-stimulated BV2 cells reduced cell viability and increased cell apoptosis in N2a cells(P<0.01).SOCS3 knockdown in BV2 cells exacerbated the reduction in N2a cell viability(P<0.01)and the increase in cell apoptosis in N2a cells(P<0.01).BAY 11-7082 treatment of these SOCS3-knockdown BV2 microglia attenuated the reduction in N2a cell viability and decreased apoptosis in N2a cells(P<0.01).Conclusion SOCS3 inhibits microglia inflammatory response through down-regulation of NF-kB activity,and in turn attenuates neuronal cell death and ameliorates PD nerve injury.

Objective To demonstrate that neferine(Nef)alleviates Parkinson's disease(PD)by inhibiting microglial pyroptosis mediated through the reactive oxygen species(ROS)/NOD-like receptor protein 3(NLRP3)/Caspase-1 pathway.Methods BV2 microglial cells were divided into:control group,lipopolysaccharides(LPS)-adenosine triphosphate(ATP)group,and LPS-ATP+Nef group.Pyroptosis was induced by 1 μg/mL LPS+5 mmol/L ATP,with 2 mmol/L Nef pretreatment.Eighteen 10-12-week-old male C57BL/6 mice(22～25 g)were randomly assigned to:control(n=6),1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)(n=6),and MPTP+Nef(n=6)groups.Detection methods included:flow cytometry for pyroptosis,Cell Counting Kit-8(CCK-8)for viability,2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)for ROS,commercial kits for malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH),ELISA/Western blot for interleukin-1β(IL-1β)/IL-18,immunofluorescence/immunohistochemistry for NLRP3/Caspase-1,tyrosine hydroxylase(TH)immunohistochemistry,hematoxylin-eosin staining for neuropathology,and modified neurological severity score(mNSS).Results Versus control,LPS-ATP group showed decreased viability(P=0.002),increased pyroptosis(P<0.001),elevated ROS(P<0.001)/MDA(P<0.001)/IL-1β(P<0.001)/IL-18(P<0.001),upregulated NLRP3(P<0.001)/Caspase-1(P<0.001),and reduced GSH(P<0.001)/SOD(P<0.001).Nef treatment reversed these effects(all P<0.05).According to the results of murine studies,compared with the control group,the MPTP group had increased mNSS(P<0.001)/tissue ROS(P<0.001),downregulated TH(P<0.001),upregulated NLRP3(P<0.001)/Caspase-1(P<0.001).Nef treatment significantly attenuated the MPTP-induced deleterious effects(P<0.05).Histopathological analysis revealed that control group exhibited uniformly distributed hippocampal neurons with distinct nuclear morphology;MPTP group showed neuronal swelling,interstitial edema,and nuclear atrophy;MPTP+Nef group demonstrated ameliorated neuronal damage.Conclusion Nef inhibits microglial pyroptosis via ROS/NLRP3/Caspase-1 axis,ameliorating PD neuroinflammation and pathology.

Objective:To evaluate the role of necroptosis in paclitaxel-induced cognitive dysfunction in mice.Methods:Thirty SPF healthy male C57BL/6N mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) using a random number table method: vehicle control group (Veh group), paclitaxel group (PTX group), and paclitaxel+ a specific inhibitor of necroptosis Necrostatin-1 group (P+ N group). In PTX group and P+ N group, paclitaxel 10 mg/kg (diluted to 5 mg/ml in anhydrous ethanol and castor oil [1∶1], and further diluted to 1 mg/ml in 0.9% normal saline before use) was intraperitoneally injected daily for 7 consecutive days to induce cognitive dysfunction. P+ N group received an intraperitoneal injection of Necrostatin-1 6.5 mg/kg (diluted to 10 mg/ml in dimethyl sulfoxide, and further diluted to 1 mg/ml in 0.9% normal saline before use) at 2 h before paclitaxel administration every other day, 4 times in total. Veh group received the equal volume of solvent at the matched time points as previously described in P+ N group. After establishment of the model, spontaneous locomotor activity was assessed using the open field test, followed by the novel object recognition test and the Morris water maze to evaluate the cognitive function. The animals were sacrificed after the end of the Morris water maze test, and the hippocampal tissues were collected for determination of the expression of necroptosis-related proteins receptor-interacting serine/threonine-protein kinase 1 (RIPK1), RIPK3, mixed lineage kinase domain-like protein (MLKL), and phospho-MLKL (p-MLKL) (by Western blot analysis) and contents of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) (double immunofluorescence staining) and for observation of the localization of programmed necrosis cells (using enzyme-linked immunosorbent assay). Results:There were no significant differences in the total distance traveled and mean movement speed in the open field test or swimming speed in the Morris water maze test among the three groups ( P>0.05). Compared to Veh group, the time spent in the central zone in the open field and time spent in the original platform quadrant were significantly shortened, the discrimination index was decreased, the escape latency was prolonged, the number of crossing the original platform was reduced, the expression of RIPK1, RIPK3, MLKL and p-MLKL was up-regulated, the contents of TNF-α and IL-1β were increased, and the number of RIPK1-positive neurons was increased in PTX group ( P<0.05). Compared to PTX group, the time spent in the central zone in the open field test and time spent in the original platform quadrant were significantly prolonged, the discrimination index was increased, the escape latency was shortened, the number of crossing the original platform was increased, the expression of RIPK1, RIPK3, MLKL and p-MLKL was down-regulated, the contents of TNF-α and IL-1β were decreased, and the number of RIPK1-positive neurons was decreased in P+ N group ( P<0.05). Conclusions:Necroptosis in hippocampal neurons can lead to neuroinflammation, thus contributeing to paclitaxel-induced cognitive dysfunction in mice.

The Janus kinase/signal transducers and activators of transcription (JAK-STAT) control natural killer (NK) cells development and cytotoxic functions, however, whether long non-coding RNAs (lncRNAs) are involved in this pathway remains unknown. We found that miR155HG was elevated in activated NK cells and promoted their proliferation and effector functions in both NK92 and induced-pluripotent stem cells (iPSCs)-derived NK (iPSC-NK) cells, without reliance on its derived miR-155 and micropeptide P155. Mechanistically, miR155HG bound to miR-6756 and relieved its repression of JAK3 expression, thereby promoting the JAK-STAT pathway and enhancing NK cell proliferation and function. Further investigations disclosed that upon cytokine stimulation, STAT3 directly interacts with miR155HG promoter and induces miR155HG transcription. Collectively, we identify a miR155HG-mediated positive feedback loop of the JAK-STAT signaling. Our study will also provide a power target regarding miR155HG for improving NK cell generation and effector function in the field of NK cell adoptive transfer therapy against cancer, especially iPSC-derived NK cells.

Objective:To investigate the clinicohistologic and molecular pathological characteristics of invasive stratified mucin-producing carcinoma(ISMC).Methods:The clinicopathological data,immunohistochemistry,alcian blue/periodic acid-Schiff(AB/PAS)staining,molecular detection and PD-L1 expressions of 11 cases of ISMC and 4 cases of stratified mucin-producing intraepithelial lesion(SMILE)in the pathological database of Anqing Medical Center of Anhui Medical University/Anqing Municipal Hospital and the first Affiliated hospital of Wannan Medical School/Yijishan Hostital between January of 2018 and March of 2024 were retrospectively analyzed.Results:ISMC patients often presented with irregular vaginal bleeding.Morphologically,the cells containing mucus in the cytoplasm were arranged in stratified layers,surrounded by a palisade,and the tumor cells might be signet ring-shaped or have clear cytoplasm.ISMC might occur in both pure and mixed forms.ISMC had highly invasive biological characteristics.CK7,p16,p40 and(or)p63 were expressed focally or positively in the form of palisades around the cancer nests.AB-PAS staining was positive.HPV test results:HPV16/18 was positive in ISMC(1/4);HPV16/18 was positive before surgery(4/6);no HPV was detected in SMILE;all ISMC cases expressed PD-L1.Eight ISMC patients were successfully followed up for 4-39 months(average 20.50 months),and four SMILE patients for 1-25 months(average 8.25 months).All followed-up patients survived;however,one ISMC patient developed multi-organ metastases post-surgery.Conclusion:ISMC has unique morphological characteristics and immunophenotypes,indicated by high invasiveness and poor prognosis.All ISMC are PD-L1 positive,suggesting that these patients may benefit from PD-L1 immunotherapy.

Objective This study aims to introduce and explore the application of the medical service price online audit system,which enables pre-positioning,informatization,intelligence,and refinement of price management,ensuring the safety of medical insurance funds and protecting the legitimate rights and interests of patients.Methods Taking a tertiary hospital in Guangdong Province as an example,the application practice of the medical institution's online audit system for price management is thoroughly introduced.Results Through the real-time monitoring of charge data by the online audit system,full-process man-agement is carried out,including pre-warning,mid-intervention,and post-analysis of medical billing behavior,in order to stand-ardize clinical diagnosis and treatment practices.Conclusion By utilizing information technology and continuously improving in-telligent audit rules,the standardization of hospital charge management can be promoted,internal control construction can be strengthened,and the hospital's refinement development can be facilitated.

Objective:To explore the correlation between the methylation of the FOS gene and the occurrence of osteoporosis (OP) in patients with gestational diabetes mellitus (GDM) .Methods:From Aug. 2017 to Aug. 2022, 120 patients with gestational diabetes mellitus (GDM) were collected in Affiliated Hospital of Jining Medical College as the research subjects. According to the bone density values, the research subjects were divided into the OP group (15 cases) and the non-OP group (105 cases) . The methylation status of the FOS gene was detected in the two groups of patients, and levels of blood calcium (Ca) , phosphorus (P) , β-collagen special sequence ( β-CTX) , procollagen I n-terminal propeptide (PINP) , osteocalcin (OC) , and bone alkaline phosphatase (AKP) were measured to analyze the correlation between FOS gene methylation and osteoporosis (OP) . Results:The frequency of FOS methylation in the OP group patients [66.67% (10/15) ] was significantly higher than that in the non-OP group patients [28.57% (30/105) ] ( χ2=8.57, P=0.003) . Compared to the non-OP group, patients in the OP group showed significant decreases in AKP and OC levels (AKP: 19.05±3.65 vs. 12.98±3.09, t=6.13, P<0.001) (OC: 12.76±1.74 vs. 9.12±1.49, t=7.70, P<0.001) , with no statistically significant differences in Ca, P, β-CTX, or PINP between the two groups ( P>0.05) . Among OP group patients, those who were FOS methylation positive showed significantly lower bone density, AKP, and OC levels compared to FOS methylation negative patients (bone density: 0.38±0.05 vs. 0.31±0.06, t=2.24, P=0.043) (AKP: 16.20±1.68 vs. 11.47±2.34, t=4.00, P=0.001) (OC: 10.27±1.55 vs. 8.58±1.14, t=2.41, P=0.032) . In Logistic multivariate analysis, FOS methylation was found to be a risk factor for osteoporosis in GDM pregnant women ( OR=1.17, 95% CI=0.04-4.26, P=0.027) , while bone density ( OR=0.71, 95% CI=0.42-2.10, P=0.012) , AKP ( OR=0.53, 95% CI=0.24-1.90, P=0.008) , and OC ( OR=0.90, 95% CI=0.17-5.18, P=0.021) were identified as protective factors for GDM patients against osteoporosis. Conclusion:The methylation of the FOS gene is significantly associated with the occurrence of OP in pregnant women with GDM, and it can increase the risk of OP in pregnant women with GDM.

Objective:To explore the rehabilitation effects of individual computer story-version magnanimous therapy (ICSMT) on patients with end-stage renal disease undergoing maintenance hemodialysis (MHD).Methods:A total of 120 patients with end-stage renal disease receiving MHD treatment at the Department of Nephrology Hemodialysis Center of Huadu District People's Hospital of Guangzhou from August 2022 to April 2024 were selected as the study subjects.They were randomly divided into control group ( n=60, receiving routine clinical treatment) and ICSMT group ( n=60, receiving routine clinical treatment combined with ICSMT for psychological intervention) by random number table method.The patients in the two groups were evaluated by the self-rating anxiety scale (SAS), self-rating depression scale (SDS), enterprising and magnanimous questionnaire (EMQ), the short-form-36 health survey (SF-36), and activity of daily living scale (ADL) before intervention and at 4-week post-intervention.Blood pressure, blood urea nitrogen (BUN), hemoglobin (Hb), and serum albumin (ALB) levels were also measured before the intervention and at the 4-week post-intervention.The clinical global impression scale (CGI) was used to evaluate clinical efficacy before the intervention, at the 2-week post-intervention, and at the 4-week post-intervention.Statistics analysis was performed using SPSS 29.0.1.0(171). Independent-samples t-test, paired t-test, Mann-Whitney U test and Wilcoxon signed-rank test were used for statistical analysis. Results:After 4 weeks of intervention, the SAS and SDS in the ICSMT group (49.0 (48.0, 50.0), 50.0 (49.0, 51.0)) were significantly lower than those in the control group (51.0 (50.0, 52.0), 52.0 (51.0, 53.0)) (both P<0.001). The enterprising subscore of the EMQ in the ICSMT group (35.0 (32.0, 37.0)) was significantly higher than that in the control group (31.0 (29.0, 34.0)) ( P<0.001). Furthermore, the differences of enterprising and magnanimous subscores between the two groups before and after intervention in the ICSMT group (2.0 (1.0, 4.0), 1.0 (-1.0, 2.0))were significantly higher than those in the control group (-1.0 (-1.0, 0), -1.0 (-1.2, 0)) (both P<0.05). Systolic and diastolic blood pressure values in the ICSMT group (130 (126, 134) mmHg, 85 (80, 88) mmHg)were significantly lower than those in the control group (145 (138, 152) mmHg, 93 (88, 99) mmHg)(1 mmHg=0.133 kPa) after 4 weeks of intervention(both P<0.05). After 4 weeks of intervention, the level of BUN in the ICSMT group (5.5 (3.7, 8.4) mmol/L) was significantly lower than that in the control group (9.1 (6.8, 11.4) mmol/L), while the level of Hb and ALB in the ICSMT group ((115.0±10.0)g/L, (38.3±3.2)g/L)were significantly higher than those in the control group ((104.0±12.0)g/L, (37.1±2.9)g/L) (all P<0.05). After 4 weeks of intervention, the physical functioning, role-physical, general health, vitality, social functioning, role-emotional, and mental health subscores of SF-36 in ICSMT group were all significantly higher than those in the control group (all P<0.05). After 4 weeks of intervention, the score of ADL in the ICSMT group (15.42±1.58)was significantly lower than that in the control group (16.78±2.06) ( t=-4.08, P<0.05). At the 2-week post-intervention and the 4-week post-intervention, the severity of illness (SI) and global improvement (GI) in the ICSMT group were significantly lower than those in the control group, while the efficacy index (EI) in the ICSMT group was significantly higher than that in the control group (all P<0.05). Conclusion:ICSMT can effectively promote the physical, psychological, and social functional rehabilitation of end-stage renal disease patients undergoing MHD, significantly improving their quality of life.

Objective:To evaluate the role of necroptosis in paclitaxel-induced cognitive dysfunction in mice.Methods:Thirty SPF healthy male C57BL/6N mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) using a random number table method: vehicle control group (Veh group), paclitaxel group (PTX group), and paclitaxel+ a specific inhibitor of necroptosis Necrostatin-1 group (P+ N group). In PTX group and P+ N group, paclitaxel 10 mg/kg (diluted to 5 mg/ml in anhydrous ethanol and castor oil [1∶1], and further diluted to 1 mg/ml in 0.9% normal saline before use) was intraperitoneally injected daily for 7 consecutive days to induce cognitive dysfunction. P+ N group received an intraperitoneal injection of Necrostatin-1 6.5 mg/kg (diluted to 10 mg/ml in dimethyl sulfoxide, and further diluted to 1 mg/ml in 0.9% normal saline before use) at 2 h before paclitaxel administration every other day, 4 times in total. Veh group received the equal volume of solvent at the matched time points as previously described in P+ N group. After establishment of the model, spontaneous locomotor activity was assessed using the open field test, followed by the novel object recognition test and the Morris water maze to evaluate the cognitive function. The animals were sacrificed after the end of the Morris water maze test, and the hippocampal tissues were collected for determination of the expression of necroptosis-related proteins receptor-interacting serine/threonine-protein kinase 1 (RIPK1), RIPK3, mixed lineage kinase domain-like protein (MLKL), and phospho-MLKL (p-MLKL) (by Western blot analysis) and contents of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) (double immunofluorescence staining) and for observation of the localization of programmed necrosis cells (using enzyme-linked immunosorbent assay). Results:There were no significant differences in the total distance traveled and mean movement speed in the open field test or swimming speed in the Morris water maze test among the three groups ( P>0.05). Compared to Veh group, the time spent in the central zone in the open field and time spent in the original platform quadrant were significantly shortened, the discrimination index was decreased, the escape latency was prolonged, the number of crossing the original platform was reduced, the expression of RIPK1, RIPK3, MLKL and p-MLKL was up-regulated, the contents of TNF-α and IL-1β were increased, and the number of RIPK1-positive neurons was increased in PTX group ( P<0.05). Compared to PTX group, the time spent in the central zone in the open field test and time spent in the original platform quadrant were significantly prolonged, the discrimination index was increased, the escape latency was shortened, the number of crossing the original platform was increased, the expression of RIPK1, RIPK3, MLKL and p-MLKL was down-regulated, the contents of TNF-α and IL-1β were decreased, and the number of RIPK1-positive neurons was decreased in P+ N group ( P<0.05). Conclusions:Necroptosis in hippocampal neurons can lead to neuroinflammation, thus contributeing to paclitaxel-induced cognitive dysfunction in mice.

Objective:To explore the correlation between the methylation of the FOS gene and the occurrence of osteoporosis (OP) in patients with gestational diabetes mellitus (GDM) .Methods:From Aug. 2017 to Aug. 2022, 120 patients with gestational diabetes mellitus (GDM) were collected in Affiliated Hospital of Jining Medical College as the research subjects. According to the bone density values, the research subjects were divided into the OP group (15 cases) and the non-OP group (105 cases) . The methylation status of the FOS gene was detected in the two groups of patients, and levels of blood calcium (Ca) , phosphorus (P) , β-collagen special sequence ( β-CTX) , procollagen I n-terminal propeptide (PINP) , osteocalcin (OC) , and bone alkaline phosphatase (AKP) were measured to analyze the correlation between FOS gene methylation and osteoporosis (OP) . Results:The frequency of FOS methylation in the OP group patients [66.67% (10/15) ] was significantly higher than that in the non-OP group patients [28.57% (30/105) ] ( χ2=8.57, P=0.003) . Compared to the non-OP group, patients in the OP group showed significant decreases in AKP and OC levels (AKP: 19.05±3.65 vs. 12.98±3.09, t=6.13, P<0.001) (OC: 12.76±1.74 vs. 9.12±1.49, t=7.70, P<0.001) , with no statistically significant differences in Ca, P, β-CTX, or PINP between the two groups ( P>0.05) . Among OP group patients, those who were FOS methylation positive showed significantly lower bone density, AKP, and OC levels compared to FOS methylation negative patients (bone density: 0.38±0.05 vs. 0.31±0.06, t=2.24, P=0.043) (AKP: 16.20±1.68 vs. 11.47±2.34, t=4.00, P=0.001) (OC: 10.27±1.55 vs. 8.58±1.14, t=2.41, P=0.032) . In Logistic multivariate analysis, FOS methylation was found to be a risk factor for osteoporosis in GDM pregnant women ( OR=1.17, 95% CI=0.04-4.26, P=0.027) , while bone density ( OR=0.71, 95% CI=0.42-2.10, P=0.012) , AKP ( OR=0.53, 95% CI=0.24-1.90, P=0.008) , and OC ( OR=0.90, 95% CI=0.17-5.18, P=0.021) were identified as protective factors for GDM patients against osteoporosis. Conclusion:The methylation of the FOS gene is significantly associated with the occurrence of OP in pregnant women with GDM, and it can increase the risk of OP in pregnant women with GDM.