OBJECTIVE: To investigate the role and mechanism of the cyclic guanosine monophosphate-adenosine monophosphate synthase/stimulator of interferon gene (cGAS/STING) pathway in oleic acid-induced acute lung injury (ALI) in mice. METHODS: Male wild-type C57BL/6J mice were randomly divided into five groups (each n = 10): normal control group, ALI model group, and 5, 50, 500 μg/kg inhibitor pretreatment groups. The ALI model was established by tail vein injection of oleic acid (7 mL/kg), while the normal control group received no intervention. The inhibitor pretreatment groups were intraperitoneally injected with the corresponding doses of cGAS inhibitor RU.521 respectively 1 hour before modeling. At 24 hours post-modeling, blood was collected, and mice were sacrificed. Lung tissue pathological changes were observed under light microscopy after hematoxylin-eosin (HE) staining, and pathological scores were assessed. Western blotting was used to detect the protein expressions of cGAS, STING, phosphorylated TANK-binding kinase 1 (p-TBK1), phosphorylated interferon regulatory factor 3 (p-IRF3), and phosphorylated nuclear factor-κB p65 (p-NF-κB p65) in lung tissue. Immunohistochemistry was performed to observe STING and p-NF-κB positive expressions in lung tissue. Serum interferon-β (IFN-β) levels were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared with the normal control group, the ALI model group exhibited significant focal alveolar thickening, intra-alveolar hemorrhage, pulmonary capillary congestion, and neutrophil infiltration in the pulmonary interstitium and alveoli, along with markedly increased pathological scores (10.33±0.58 vs. 1.33±0.58, P < 0.05). Protein expressions of cGAS, STING, p-TBK1, p-IRF3, and p-NF-κB p65 in lung tissue significantly increased [cGAS protein (cGAS/β-actin): 1.24±0.02 vs. 0.56±0.02, STING protein (STING/β-actin): 1.27±0.01 vs. 0.55±0.01, p-TBK1 protin (p-TBK1/β-actin): 1.34±0.03 vs. 0.22±0.01, p-IRF3 protein (p-IRF3/β-actin): 1.23±0.02 vs. 0.36±0.01, p-NF-κB p65 protein (p-NF-κB p65/β-actin): 1.30±0.02 vs. 0.53±0.02, all P < 0.05], positive expressions of STING and p-NF-κB in lung tissue were significantly elevated [STING (A value): 0.51±0.03 vs. 0.30±0.07, p-NF-κB (A value): 0.57±0.05 vs. 0.31±0.03, both P < 0.05], and serum IFN-β levels were also significantly higher (ng/L: 256.02±3.84 vs. 64.15±1.17, P < 0.05). The cGAS inhibitor pretreatment groups showed restored alveolar structural integrity, reduced inflammatory cell infiltration, and decreased hemorrhage area, along with dose-dependent lower pathological scores as well as the protein expressions of cGAS, STING, p-TBK1, p-IRF3 and p-NF-κB p65 in lung tissue, with significant differences between the 500 μg/kg inhibitor group and ALI model group [pathological score: 2.67±0.58 vs. 10.33±0.58, cGAS protein (cGAS/β-actin): 0.56±0.03 vs. 1.24±0.02, STING protein (STING/β-actin): 0.67±0.03 vs. 1.27±0.01, p-TBK1 protein (p-TBK1/β-actin): 0.28±0.01 vs. 1.34±0.03, p-IRF3 protein (p-IRF3/β-actin): 0.32±0.01 vs. 1.23±0.02, p-NF-κB p65 protein (p-NF-κB p65/β-actin): 0.63±0.01 vs. 1.30±0.02, all P < 0.05]. Compared with the ALI model group, positive expressions of STING and p-NF-κB in lung tissue were significantly reduced in the 500 μg/kg inhibitor group [STING (A value): 0.40±0.01 vs. 0.51±0.03, p-NF-κB (A value): 0.43±0.02 vs. 0.57±0.05, both P < 0.05], and serum IFN-β levels were also markedly reduced (ng/L: 150.03±6.19 vs. 256.02±3.84, P < 0.05). CONCLUSIONS The cGAS/STING pathway is activated in oleic acid-induced ALI, leading to exacerbated inflammatory responses and increased lung damage. RU.521 can inhibit cGAS, thereby down-regulating the expression of pathway proteins and cytokines, and providing protection to lung tissue.
Animals ; Acute Lung Injury/chemically induced* ; Male ; Nucleotidyltransferases/metabolism* ; Mice ; Signal Transduction ; Mice, Inbred C57BL ; Membrane Proteins/metabolism* ; Oleic Acid/adverse effects* ; Transcription Factor RelA/metabolism* ; Lung/pathology* ; Interferon Regulatory Factor-3/metabolism* ; Disease Models, Animal

Objective:To investigate the current status of activation in patients with spinal cord injury and analyze its influencing factors.Methods:Using the convenient sampling method, a total of 127 patients from Department of Spinal Surgery in the Third Affiliated Hospital of Soochow University from October 2020 to August 2021 were selected as the research objects. The patients were investigated by General Data Questionnaire, Patient Activation Measure (PAM) , General Self-Efficacy Scale (GSES) , Connor-Davidson Resilience Scale (CD-RISC) , Social Support Rating Scale (SSRS) and Hospital Anxiety and Depression Scale (HADS) . Multiple linear hierarchical regression analysis was used to investigate the influencing factors of activation in patients with spinal cord injury.Results:A total of 127 questionnaires were sent out, and 122 were effectively received, with the effective recovery of 96.1%. The score of activation in patients with spinal cord injury was (54.71±7.23) . Regression analysis showed that injury site, mental resilience and social support were the main influencing factors of activation ( P<0.05) , and the global regression model explained 50.0% of the variation in activation in patients with spinal cord injury. Conclusions:Patients with spinal cord injury generally have moderate levels of activation during hospitalization. The activation of patients with spinal cord injury is a positive psychosocial resource, and rehabilitation nurses should pay more attention to it, and improve the mental resilience and social support of patients, so as to further enhance the activation of patients.

Objective To view and compare the clinical characteristics of renal tubular acidosis in adults and children.Methods Clinical data of patients with renal tubular acidosis diagnosed by Shandong Provincial Hospital affiliated to Shandong University from Jan 1991 to Sep 2017 were reviewed.The difference and consistency in clinical characteristics of renal tubular acidosis between adults and children were analyzed.Results Data from 206 adults and 60 children were analyzed.89.81％ cases in adults were secondary to other diseases,mainly primary Sjogren's syndrome.Most children patients (81.67％) were idiopathic,others largely originated from inherited metabolic diseases.The most common subtype of both was distal renal tubular acidosis.Proximal renal tubular acidosis was easier to be found in idiopathic renal tubular diseases of children.Chief complaints or starting symptoms were mainly composed of polydipsia with polyuria (41.4％) and fatigue (35.3％).Children were typical of growth retardation,rickets and digestive symptoms.The rate of missed diagnosis and misdiagnosis was 41.4 percent.Routine therapy consisted of healing metabolic acidosis and electrolyte disorders,treating underlying diseases and preventing complications.The majority of patients (95.5％) improved after treatments.Conclusions Renal tubular acidosis possesses various underlying diseases,diverse clinical manifestation and high rate of misdiagnosis.Given the high incident of secondary types,investigation of underlying disease,especially autoimmune diseases such as Sjogren's syndrome,is of great importance in adults.Most children patients suffer from primary renal tubular acidosis.Attention should be paid to them in order to reduce the rate of misdiagnosis and teratogenicity.

Objective To deveplope construct and validate a novel multiplex PCR system comprised of 30 Y-STR markers only with low and moderate mutation rates. Methods 30 Y-STRs characterized by low/moderate mutation rate and middle/high polymorphic was amplified simultaneously in a multiplex PCR system using the six color labeling fluorescence. PCR product was analyzed in a ABI 3500XL Genetic Analyzer. The accuracy, specifity, sensitivity and stability of the system and its validation on the mixtures were evaluated. Results The validation studies demonstrated that the system is a stable, accurate, and sensitive multiplex PCR system. The sensitivity was 0.0625ng DNA. Y-STR could be detection in a male/female DNA mixture ratio of 1:4. Conclusion The primary study demonstrates that this multiplex PCR system is effective and reliable for forensic routine DNA analysis. It will be very helpful for constructing Chinese forensic Y-STR database and population genetic research.

Objective: To investigate the expressions of migration and invasion inhibitory protein (MIIP) and p21-activated kinase 1 (PAK1) in endometrial carcinoma (EC) and their correlation with clinicopathological features. Methods: The protein levels of MIIP and PAK1 in 135 paraffin-embedded EC tissues, 55 atypical hyperplasia of endometrium (AHE) and 88 normal endometrium (NE) tissues were quantified by immunohistochemistry, the clincial significance and the relationship of these two proteins were also analyzed. Results: The positive rates of MIIP expression in NE, AHE and EC tissues were 52.3%(46/88), 41.8% (23/55) and 34.8% (47/135), respectively. The expression of MIIP in EC was significantly lower than that of MIIP in NE (P<0.05). The positive rates of PAK1 expression in NE, AHE and EC tissues were 45.5% (40/88), 50.9% (28/55) and 62.2% (84/135), respectively. The expression of PAK1 in EC tissues was significantly higher than that of PAK1 in NE tissues (P<0.05). The expression of MIIP in EC tissues was significantly associated with myometrial invasion, International Federation of Gynaecology and Obstetrics (FIGO) stage and lymph node metastasis (P<0.05). The expression of PAK1 in EC tissues was significantly related with differentiation, myometrial invasion, FIGO stage and lymph node metastasis (P<0.05). The expressions of MIIP and PAK1 in EC tissues were marginally related with the overall survival of patients (P=0.092, P=0.052). The expression of MIIP in EC was negatively correlated with PAK1 (r=-0.329, P<0.001). Conclusions The down-regulation of MIIP and up-regualtion of PAK1 paticipate in the initiation and development of EC, which are correlated with the poor prognosis of EC. The protein expression of MIIP is inversely related with PAK1 in EC.

Objective To explore the protective effect of miconazole on white matter damage (WMD) in neonatal rats. Methods Three-day-old neonatal SD rats were randomly divided into sham group, WMD model group, 10 mg/(kg·d) miconazole group and 40 mg/(kg·d) miconazole group with 15 rats each. Rats in WMD model group were subjected to the ligation of right carotid artery, and then kept in a chamber with 6% oxygen and 94% nitrogen for 80 min to establish the white matter damage model. The rats in miconazole group were intraperitoneally injected with different doses (10 and 40mg/kg) of miconazole, dissolved in dimethyl sulfoxide (DMSO), for five consecutive days, and rats in WMD model group were injected with the same volume of DMSO. Myelin basic protein (MBP) of white matter was detected by immunofluorescence staining and western blot. Myelin sheaths of corpus callosum were observed by transmission electron microscopy. Weight changes of rats were compared among groups. Results Immunofluorescence staining and western blot showed that, after treatment with miconazole, the MBP expression level of corpus callosum was higher than in WMD model group (P<0.05). In WMD model group, the myelin sheath of corpus callosum had loose structure and a large number of small vacuoles with decreased thickness of myelin sheath. After treatment with miconazole, myelinolysis induced by anoxia and ischemia could be improved significantly. The increase in weight of rats in WMD model group was significantly less than that in sham group. And after miconazole treatment, the rate of weight gain of rats was increased. Conclusion Miconazole can significantly reduce the brain white matter damage induced by anoxia and ischemia through promoting myelination, and then improves the growth and development in rats.

Objective The aim of this study was to investigate mutations of 41 STR loci. Methods 4546 bloodstain samples were typed from 1932 father–mother–child trios by using AGCU_21+1, AGCU_EX22 and GlobalFiler_ExpressTM amplification Kit. Calculate the mutation rates of STR loci. Results 154 mutations were identified at 32 of the 41 loci. The average mutation rate was 1.0×10-3per locus(95%CI: 0.8~1.1×10-3), and the mutations of SE33 was highest. 152(98.7%) mutation events were one-step mutation, 2(1.3%) events were two-steps. The mutation events occurred in 150 father–mother–child triplets. The mutations in 146(97.3%) triplets occurred at single locus, 8 mutations were observed at two loci in 4(2.7%) triplets simultaneously. 104 paternal and 22 maternal mutations could be determined under 79212 paternal and maternal allelictransfers. The ratio of paternal versus maternal mutations was 4.7:1, and 28 unassigned mutations were observed. Conclusion STR mutation are common in paternity testing, and we should pay more attention to it.

Objective To compare the effect of silica-extraction method and Silico membrane based method in DNA purification from bones and teeth.Methods DNA samples were purified respectively with the silica-extraction method and MinElute PCR Purification kit from 6 bones and 8 teeth,then tested STR types by GlobalFiler? kits. And evaluated the two methods with the success rate and the peak height. Results Both of the two purification methods can successfully obtain the STR markers of the 14 samples. And there was no statistical difference between the two methods in the average peak height from bones and teeth. Conclusion The Silico membrane based method which have more advantages in operation is an efficient method to purify DNA from bones and teeth, and there is no significant difference compared with the silica-extraction method. But the cost is higher. It can be selectively used in forensic practice.

Objective To provide the basic data for forensic application by analyzing the genotypes absence at DYS448 locus. Method 5487 bloodstain samples from unrelated male individuals of Chinese Han population were obtained. 4479 samples was co-amplified using Y-filerTM and AGCU Y-24 kits. 1008 samples was co-amplified using Yfiler PlusTM and AGCU Y-24 kits .Probability of genotype absence was calculated. Results 35 samples of 35 haplotypes among 5487 non-related individuals were found to have DYS448 genotypes absence ,while 2 individuals displayed additional alleles at else locus. Conclusion The probability of DYS448 genotype absence was 0.637%, forensic scientists should pay more attention in practical cases and YSTR database.