Noise-induced hearing loss is a worldwide public health issue that is characterized by temporary or permanent changes in hearing sensitivity. This condition is closely linked to inflammatory responses, and interventions targeting the inflammatory gene tumor necrosis factor-alpha (TNFα) are known to mitigate cochlear noise damage. TNFα-induced proteins (TNFAIPs) are a family of translucent acidic proteins, and TNFAIP6 has a notable association with inflammatory responses. To date, there have been few reports on TNFAIP6 levels in the inner ear. To elucidate the precise mechanism, we generated transgenic mouse models with conditional knockout of Tnfaip6 (Tnfaip6 cKO). Evaluation of hair cell morphology and function revealed no significant differences in hair cell numbers or ribbon synapses between Tnfaip6 cKO and wild-type mice. Moreover, there were no notable variations in hair cell numbers or hearing function in noisy environments. Our results indicate that Tnfaip6 does not have a substantial impact on the auditory system.
Animals ; Mice, Knockout ; Hair Cells, Auditory, Inner/pathology* ; Mice ; Mice, Transgenic ; Hearing Loss, Noise-Induced ; Evoked Potentials, Auditory, Brain Stem/physiology*

Objective:To analyze the carriage status, subtype distribution and flanking gene sequence characteristics of oxacillinases (OXA enzyme) in 241 clinical strains of Pseudomonas aeruginosa, and assess their roles in the drug resistance of Pseudomonas aeruginosa and ability to horizontally transfer across species. Methods:Clinical P. aeruginosa isolates were collected from four hospitals in Sanya, Tangshan, Zhangjiakou, and Beijing. The prevalence of oxacillinases and their flanking gene sequences was analyzed by whole-genome sequencing (NGS) and bioinformatic approaches. Results:A total of 241 isolates of P. aeruginosa were gathered, and 35 blaOXA subtypes were identified through screening of 252 blaOXA genes. These genes were classified into three subfamilies: blaOXA-50-like (241, 95.6%), blaOXA-1-like (9, 3.6%) and blaOXA-10-like (2, 0.8%). Among these, 11 subtypes (11, 31.4%) were novel blaOXA subtypes. Nine of these belonged to the blaOXA-50-like subfamily and were designated as blaOXA-1244, blaOXA-1245, blaOXA-1246, blaOXA-1250, blaOXA-1252, blaOXA-1253, blaOXA-1254, blaOXA-1255, and blaOXA-1256. The remaining two belonged to the blaOXA-10-like subfamily and were named blaOXA-1247 and blaOXA-1248. Compared to the amino acid sequence of OXA-10, the newly identified subtype OXA-1247 exhibited a mutation at position 117, where a valine was replaced by a leucine. This change was thought to improve the enzyme′s ability to hydrolyze carbapenems. In the analysis of the flanking sequences of the blaOXA genes, Class I integrons were identified in four bacterial strains. The variable regions of these integrons carried three distinct patterns of resistance gene cassettes: aac( 6′) -Ib-blaOXA-1247-ant( 3′′) -Ia, aac( 6′) -Ib-blaOXA-1248 and aac( 6′) -Ib- blaIMP-45-blaOXA-1-catB3. Among these, the strain BJ2326 carried a class I integron that was connected to the downstream IS CR1 element to form a composite class I integron structure, additionally carrying the resistance gene blaPER-1. Out of the 223 non-wild-type P. aeruginosa strains, 127 strains exhibited non-wild-type profiles to the four beta-lactam antibiotics MEM, CAZ, FEP, and TZP, with the combination of MEM+CAZ+FEP being the most prevalent, representing 57.0% of the total. Conclusions:The blaOXA genes in 241 clinical P. aeruginosa strains showed diversity. Some blaOXA genes had a co-transfer risk with the metallo-β-lactamase resistance gene blaIMP-45. Among the 11 newly discovered blaOXA subtypes, the new subtype OXA-1247 may have carbapenemase activity and potential for horizontal transfer.

Objective:To analyze the carriage status, subtype distribution and flanking gene sequence characteristics of oxacillinases (OXA enzyme) in 241 clinical strains of Pseudomonas aeruginosa, and assess their roles in the drug resistance of Pseudomonas aeruginosa and ability to horizontally transfer across species. Methods:Clinical P. aeruginosa isolates were collected from four hospitals in Sanya, Tangshan, Zhangjiakou, and Beijing. The prevalence of oxacillinases and their flanking gene sequences was analyzed by whole-genome sequencing (NGS) and bioinformatic approaches. Results:A total of 241 isolates of P. aeruginosa were gathered, and 35 blaOXA subtypes were identified through screening of 252 blaOXA genes. These genes were classified into three subfamilies: blaOXA-50-like (241, 95.6%), blaOXA-1-like (9, 3.6%) and blaOXA-10-like (2, 0.8%). Among these, 11 subtypes (11, 31.4%) were novel blaOXA subtypes. Nine of these belonged to the blaOXA-50-like subfamily and were designated as blaOXA-1244, blaOXA-1245, blaOXA-1246, blaOXA-1250, blaOXA-1252, blaOXA-1253, blaOXA-1254, blaOXA-1255, and blaOXA-1256. The remaining two belonged to the blaOXA-10-like subfamily and were named blaOXA-1247 and blaOXA-1248. Compared to the amino acid sequence of OXA-10, the newly identified subtype OXA-1247 exhibited a mutation at position 117, where a valine was replaced by a leucine. This change was thought to improve the enzyme′s ability to hydrolyze carbapenems. In the analysis of the flanking sequences of the blaOXA genes, Class I integrons were identified in four bacterial strains. The variable regions of these integrons carried three distinct patterns of resistance gene cassettes: aac( 6′) -Ib-blaOXA-1247-ant( 3′′) -Ia, aac( 6′) -Ib-blaOXA-1248 and aac( 6′) -Ib- blaIMP-45-blaOXA-1-catB3. Among these, the strain BJ2326 carried a class I integron that was connected to the downstream IS CR1 element to form a composite class I integron structure, additionally carrying the resistance gene blaPER-1. Out of the 223 non-wild-type P. aeruginosa strains, 127 strains exhibited non-wild-type profiles to the four beta-lactam antibiotics MEM, CAZ, FEP, and TZP, with the combination of MEM+CAZ+FEP being the most prevalent, representing 57.0% of the total. Conclusions:The blaOXA genes in 241 clinical P. aeruginosa strains showed diversity. Some blaOXA genes had a co-transfer risk with the metallo-β-lactamase resistance gene blaIMP-45. Among the 11 newly discovered blaOXA subtypes, the new subtype OXA-1247 may have carbapenemase activity and potential for horizontal transfer.

Objective:To explore the effect of enriched environment on the A1/A2 phenotype conversion of astrocytes and cognitive function in rats after ischemia-reperfusion (I/R) in rats. Method:Forty two adult male SD rats (weight 220±20g) were selected for middle cerebral artery occlusion (MCAO) surgery,followed by reperfusion 2 hours later. Three days after operation,30 rats were randomly di-vided into standard environment group (n=15) and enriched environment group (n=15),and 10 rats were se-lected as sham operation group. The enriched environment group was raised in the enriched environment cag-es,and the other two groups were raised in the standard environment. After 21 days,the Bederson score and mNss score were used to detect the behavioral changes of rats in each group,and the Morris water maze was used to detect the cognitive function of rats. Subsequently,western blot analysis was used to analyze the acti-vation of glial fibrillary acidic protein (GFAP),a marker of astrocytes. Real-time PCR and ELISA detect the expression of A1 type astrocyte marker (C3) and A2 type astrocyte marker (S100A10). Hematoxylin eosin (HE) staining was used to observe the pathological changes of cortex around the infarction. TUNEL staining was used to detect apoptosis.Result:Compared with SE group,the expression of GFAP protein in EE group decreased significantly(P<0.05). The expression level of the A1 type astrocyte marker C3 in EE group was significantly lower than that in SE group (P<0.05),while the expression of A2 type astrocyte marker S100A10 was significantly higher than that in SE group (P<0.05). Correspondingly to this result,in the EE group,the secretion of the cyto-kine TNF-α by A1 astrocytes was significantly lower than in the SE group(P<0.05),and the secretion of the cytokine BDNF by A2 astrocytes was significantly higher than in the SE group (P<0.05). TUNEL and HE staining showed that the apoptosis and damage of cells in EE group were less (P<0.05). In addition,the neu-rological ischemia in the EE group was less severe,including significant differences in the Bederson score and mNss score (P<0.01). Compared with the SE group,the rats in the EE group had better cognitive func-tion,characterized by shorter latency (P<0.05),longer residence time in the target quadrant (P<0.01),and more times of crossing the platform (P<0.05) in the water maze experiment.Conclusion:The enriched environment can inhibit the activation of astrocytes,promote the conversion of acti-vated astrocytes to neuroprotective type A2 and inhibit their conversion into neurotoxic type A1,resulting in improving cognitive function after ischemic stroke in rats.

Objective:To explore the effect of enriched environment on the A1/A2 phenotype conversion of astrocytes and cognitive function in rats after ischemia-reperfusion (I/R) in rats. Method:Forty two adult male SD rats (weight 220±20g) were selected for middle cerebral artery occlusion (MCAO) surgery,followed by reperfusion 2 hours later. Three days after operation,30 rats were randomly di-vided into standard environment group (n=15) and enriched environment group (n=15),and 10 rats were se-lected as sham operation group. The enriched environment group was raised in the enriched environment cag-es,and the other two groups were raised in the standard environment. After 21 days,the Bederson score and mNss score were used to detect the behavioral changes of rats in each group,and the Morris water maze was used to detect the cognitive function of rats. Subsequently,western blot analysis was used to analyze the acti-vation of glial fibrillary acidic protein (GFAP),a marker of astrocytes. Real-time PCR and ELISA detect the expression of A1 type astrocyte marker (C3) and A2 type astrocyte marker (S100A10). Hematoxylin eosin (HE) staining was used to observe the pathological changes of cortex around the infarction. TUNEL staining was used to detect apoptosis.Result:Compared with SE group,the expression of GFAP protein in EE group decreased significantly(P<0.05). The expression level of the A1 type astrocyte marker C3 in EE group was significantly lower than that in SE group (P<0.05),while the expression of A2 type astrocyte marker S100A10 was significantly higher than that in SE group (P<0.05). Correspondingly to this result,in the EE group,the secretion of the cyto-kine TNF-α by A1 astrocytes was significantly lower than in the SE group(P<0.05),and the secretion of the cytokine BDNF by A2 astrocytes was significantly higher than in the SE group (P<0.05). TUNEL and HE staining showed that the apoptosis and damage of cells in EE group were less (P<0.05). In addition,the neu-rological ischemia in the EE group was less severe,including significant differences in the Bederson score and mNss score (P<0.01). Compared with the SE group,the rats in the EE group had better cognitive func-tion,characterized by shorter latency (P<0.05),longer residence time in the target quadrant (P<0.01),and more times of crossing the platform (P<0.05) in the water maze experiment.Conclusion:The enriched environment can inhibit the activation of astrocytes,promote the conversion of acti-vated astrocytes to neuroprotective type A2 and inhibit their conversion into neurotoxic type A1,resulting in improving cognitive function after ischemic stroke in rats.

ObjectiveTo investigate the intestinal absorption characteristics of multi-index components in Danggui Buxuetang with drug absorption simulating system （DASS） established by everted intestinal sac model. MethodThe intestinal absorption solution at different time points after administration of Danggui Buxuetang was collected and detected by high performance liquid chromatography （HPLC）， acetonitrile （A）-0.2% glacial acetic acid solution （B） was used as the mobile phase for gradient elution （0-16 min， 15%-23%A； 16-20 min， 23%-28%A； 20-25 min， 28%-30%A； 25-30 min， 30%A； 30-35 min， 30%-65%A； 35-45 min， 65%-95%A）， the detection wavelength was 302 nm. HPLC fingerprint of intestinal absorption solution was established and the common peak was calibrated， and the relative cumulative absorption rate of each index component was calculated. The relative cumulative absorption curves of components were fitted with various mathematical models by DDSolver 1.0 to explore the absorption law of different components. ResultThe absorption process of C2 （calycosin-7-glucoside） and C6 in Danggui Buxuetang was in line with zero-order equation， C9 was best fitted by Weibull equation， and the remaining 7 components were in line with Makoid-Banakar equation. C1 with C2， C3， C5， C7 and C10， C2 with C5 and C7， C3 with C4， C5， C7 and C10， C4 with C6 and C10， C5 with C7， C6 with C10， C7 with C10， C8 with C9 were absorbed simultaneously during the absorption process. With the prolongation of time， the overall cumulative absorption rate of Danggui Buxuetang increased. At 120 min， the overall cumulative absorption rate of Danggui Buxuetang exceeded 38%， and reached 49.14% at 180 min. ConclusionTen ingredients in Danggui Buxuetang are absorbed in the jejunum， but absorption law of various components is different， which shows that the intestinal absorption of compound preparations of traditional Chinese medicine （TCM） has multiple characteristics. Intestinal absorption study of TCM compound preparations with chemical composition as the index can reveal some of its absorption law， but it is not complete.

Objective To investigate clinical and myopathological features and genetic mutations of GNE myopathy.Methods The clinical manifestations and pathological findings of 4 patients were reviewed who had been treated for GNE myopathy at Department of Neurology,The People's Hospital of Zhengzhou University from January 2016 to July 2017.The exons of GNE gene were sequenced to detect gene mutations in all of them.Results Onset of the disease started at late adolescence or early adulthood in the 4 patients (2 siblings).Gait disturbance appeared as an initial symptom.The disease progressed slowly and the distal muscles were more affected than the proximal ones.Muscle magnetic resonance imaging revealed that the soleus and the anterolateral muscles of the lower leg were the most severely involved and the internal and posterior compartments of the thigh muscles were more involved than the anterior one.The characteristic pathological finding was presence of rimmed vacuoles.Two siblings (Patients 1 and 2) both had the same compound heterozygous mutations of p.G395R and p.D207V,patient 3 had the compound heterozygous mutations of p.D207V and p.G166D,and patient 4 had the homozygous mutation ofp.G237D.Conclusions Sparing of the quadriceps relatively is a very unique feature of GNE myopathy.Missense mutations (G395R and G237D) may be 2 new ones in the GNE gene.

Objective:To investigate the expression of transmembrane 4 super family 1 (TM4SF1)in breast cancer tissue,and to elucidate its clinical significance and explore the related molecular biological mechanisms. Methods:A total of 190 cases of human breast cancer,110 cases of paracancerous tissue and 110 cases of normal breast tissue were collected.Immunohistochemistry was used to detect the expression levels of TM4SF1 mRNA in breast cancer tissue,paracancerous tissue,and normal breast tissue;Western blotting method was used to detect the expression levels of TM4SF1 in breast cancer tissue,paracancerous tissue,and normal breast tissue;RT-PCR method was used to detect the expression levels of TM4SF1 mRNA in breast cancer tissue,paracancerous tissue, and normal breast tissue.The positive expression rates of TM4SF1 in breast cancer tissue of the breast cancer patients with different clinicopathological features were detected.Results:The positive expression rate of TM4SF1 in the breast cancer tissue was significantly higher than those in paracancerous tissue and normal breast tissue (P <0.05);there was no significant difference in the positive expression rates of TM4SF1 between paracancerous tissue and normal breast tissue (P = 0.531);the expression of TM4SF1 was not correlated with age,but was closely correlated with tumor size,differentiation degree,lymph node metastasis and tumor stage (P <0.05);the positive expression rate of TM4SF1 in basal like breast cancer tissue was higher than those in the other three types of tissues (P <0.05).The results of Western blotting showed that the expression level of TM4SF1 in breast cancer tissue was higher than those in paracancerous tissue and normal breast tissue (P < 0.05 ), but there was no significant difference in the expression level of TM4SF1 between paracancerous tissue and normal breast tissue (P >0.05). The results of RT-PCR showed that the expression level of TM4SF1 mRNA in breast cancer tissue was higher than those in the paracancerous tissue and normal breast tissue (P <0.01);there was no significant difference in the expression level of TM4SF1 mRNA between paracancerous tissue and normal breast tissue (P > 0.05 ). Conclusion:The TM4SF1 is highly expressed in breast cancer tissue. TM4SF1 may affect the occurrence, development and distant metastasis of breast cancer through various mechanisms.TM4SF1 may be a potential target for the treatment of breast cancer.

Objective To analyze the clinical characteristics,imaging manifestations and prognoses of anti-GABAB receptor antibodies encephalitis.Methods The clinical manifestations,laboratory findings and radiological data of 13 patients with anti-GABAB receptor encephalitis,admitted to our hospital from September 2015 to March 2017,were retrospectively analyzed.Modified Rankin scale (mRs) was performed to evaluate the prognoses (good prognosis:mRs scores ＜ 2;poor prognosis mRs scores≥3).Results These 13 patients had an average age of 58 years (ranged 49-76 years) with a male to female ratio of 12:1.The major clinical features,including epileptic seizure,were found in 12 patients,psychiatric symptoms in 11 patients,cognitive disorder in 7 patients,and disturbance of consciousness in 4 patients.Brain MR imaging showed abnormal signal in 5 patients:4 were located in the hippocampus and amygdaloid,and one in the pons and left temporal lobe.Five patients showed abnormalities in PET-CT,including 4 with temporal hypermetabolism and 1 with cortical hypometabolism.Chest CT showed lung occupying lesions in 4 patients,of which 2 patients were diagnosed as having small cell lung cancer (SCLC) by pathological examination.Ten patients received immunomodulatory therapy,and three were with supportive care.After the average of 8 months of follow-up,7 patients had good prognosis,5 patients had poor prognosis and one patient lost of follow up.Conclusions Anti-GABAB receptor encephalitis frequently occurs in elderly male subjects and the main characteristic includes prominent refractory epilepsy and shows neurological improvement on immunotherapy.It can accompany by SCLC and have a relatively poor prognosis.

Objective To explore the clinical significance of expressing multiple autoantibodies in patients with autoimmune encephalitis.Methods Cerebrospinal fluid and serum were tested in patients with undefined encephalitis admitted to Peking Union Medical College Hospital from May 2013 to December 2014.Indirect immunofluorescence test was firstly used to identify the antibodies to neuronal cell-surface or synaptic receptors (including N-methyl-D-aspartate receptor (NMDAR),contactin-associated protein-like 2 (CASPR2),α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR),leucine-rich glioma inactivated protein 1 (LGI1),and gamma-aminobutyric acid beta receptor (GABABR)).In those patients with positive antibodies,antibodies against intracellular neuronal antigens associated with paraneoplastic neurological symptoms were tested.Anti-aquaporin protein-4 (AQP4) antibody was tested depending on patients' clinical manifestations.Results Ten patients were detected combined with additional autoantibodies in 531 patients with positive antibodies related to autoimmune encephalitis.AntiHu antibody was positive in 5 patients with anti-GABABR encephalitis,in 1 of whom anti-NMDAR antibody was also identified;anti-AQP4 antibody was positive in 1 patient with relapsing anti-NMDAR encephalitis;anti-CASPR2 and anti-Yo antibodies were respectively positive in 2 patients with anti-LGI1 encephalitis;anti-CV2 and anti-Hu antibodies were respectively positive in 2 patients with anti-AMPAR encephalitis.Clinical presentation of all cases was consistent with typical encephalitis or limbic encephalitis.Brain stem was involved in 3 patients.Peripheral sensory neuropathy was present in 1 patient,while myalgia and fasciculation were present in 1 patient.Seven patients responded well to the immunotherapy.Tumors were pathologically or radiologically confirmed in 7 cases,including lung cancer in 5 cases,suspected thymoma in 1 case and highly suspected mediastinal tumor without pathological identification in 1 case.Conclusions Due to the pathological mechanism,co-existence of multiple autoantibodies affects clinical manifestations of patients and results in variation and overlap of them.The additional positivity of onconeuronal antibodies directs the search for occult tumor.