Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.‍T260A, p.‍R422W, and p.‍R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‍‒‍49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‍‒‍17.9%, which was significantly higher than that (6.9%‍‒‍7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.
Humans ; Apoptosis Inducing Factor/metabolism* ; NAD/metabolism* ; Dimerization ; Apoptosis

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15％ of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.T260A, p.R422W, and p.R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5％?49.7％ of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3％?17.9％, which was significantly higher than that (6.9％?7.4％) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.

Objective:To investigate the techniques used in blood flow control of Kimura laparoscopic spleen-preserving pancreatectomy (LSPDP).Methods:Forty·five patients with benign or low-grade malignant pancreatic diseases undergoing LSPDP at Huzhou Central Hospital from May 2014 to Oct 2021 were analyzed retrospectively. Patients were divided into splenic vascular flow control group ( n=22) and routine management group ( n=23). Results:There was no significant difference in gender, age, BMI, accompanying symptoms, hypertension, diabetes, lesion size and pathological diagnosis between the two groups (all P>0.05). A higher overall spleen preservation rate (90.9% vs. 52.2%, χ2=8.213, P=0.004), lower incidence of morbidity with Clavien grade ≥ Ⅱ (22.7% vs. 73.9%, χ2=9.911, P=0.002) and shorter postoperative hospital stay [(9.6±4.5) d vs. (14.3±6.6) d, t=2.447, P=0.008] were achieved in the vascular flow control group compared with those in the routine group. Conclusion:Splenic vascular flow control techniques improve the success rate of spleen preservation in laparoscopic distal pancreatectomy, reduce the postoperative complications and shorten the postoperative hospital stay.

Objective:To study the changes of cytokines after cardiopulmonary bypass(CPB)in children with congenital heart disease.Methods:A total of 124 children with congenital heart disease underwent CPB surgery at Shanghai Children′s Medical Center from June 2020 to October 2021 with cytokine detection were enrolled.Twelve kinds of cytokines, white blood cell count(WBC)and neutrophil-to-lymphocyte ratio(NLR), C-reactive protein(CRP)and procalcitonin were detected before and 24 hours after operation.All patients were divided into CPB<120 min group ( n=102)and CPB≥120 min group ( n=22)acoording to CPB time, and were divided into systemic inflammatory response syndrome (SIRS) group, compensatory anti-inflammatory response syndrome (CARS) group and control group according to the changes of cytokines.The changes of cytokines, anti-inflammatory factors and pro-inflammatory factors before and after CPB and the correlation with CPB time were analyzed. Results:There were 65 boys and 59 girls with a body weight of(10.69±8.18)kg and a median age of 317(141, 975)d.After CPB, WBC(×10 9/L)(13.47 vs.8.6), NLR(4.93 vs.0.55), and CRP(mg/L)(81.35 vs.0.8) were significantly higher than those before operation( P<0.001). IL-6(pg/mL)(135.69 vs.6.86), IL-8(pg/mL)(33.33 vs.14.95), and IL-10(pg/mL)(6.05 vs.2.44)were significantly higher than those before operation( P<0.001). Compared with CPB<120 min group, IL-6(pg/mL)(211.88 vs.119.47), IL-8(pg/mL)(71.67 vs. 25.39), and IL-10(pg/mL)(7.69 vs. 4.92)in CPB≥120 min group significantly increased( P<0.001). CRP was negatively correlated with CPB time( r=-0.204, P=0.025), while IL-6( r=0.254, P=0.005), IL-8( r=0.358, P=0.001), IL-10( r=0.198, P=0.03) were positively correlated with CPB time.Twelve children(9.7%)had obvious SIRS, and four cases(3.2%)had early CARS.The mortality of CARS group was significantly higher than that of SIRS group and the control group( P=0.011). Conclusion:Il-6 , IL-8, and IL-10 are significantly increased after CPB in children with congenital heart disease.With the increase of CPB time, IL-6 and IL-8 increase significantly, and the correlation between IL-8 and CPB time is the strongest.Although the proportion of children with early postoperative CARS is small, the mortality is high, which indicates clinical surveillance and treatment need to be strengthened for anti-inflammatory response.

Objective:To explore the curative effect of synchronous bionic electrical stimulation in hyperbaric oxygen chamber on patients with sudden deafness（SD）and the changes in hemorheological indicators.Methods:A total of 128 SD outpatients and inpatients treated at the Hyperbaric Oxygen Department of the Sixth Medical Center of the Chinese PLA General Hospital from June 2020 to June 2021 were randomly divided into study group（70 cases treated with synchronous brain bionic electrical stimulator in hyperbaric oxygen chamber）and control group（58 cases given conventional hyperbaric oxygen therapy），respectively. The curative effects of the two groups were evaluated，and the hemorheological indicators，such as the whole blood（high-shear and low-shear）viscosity，plasma viscosity，and red blood cell aggregation，were analyzed by the automatic biochemical analyzer before and after treatment.Results:After two courses of treatments，the overall effective rate of the study group（88.57%）was significantly higher than that of the control group（79.31%），and the difference was statistically significant（ P<0.05）. After treatment，the whole blood（high-shear and low-shear）viscosities，plasma viscosity，and erythrocyte aggregation indicators of the two groups were significantly decreased compared with those before treatment（ P<0.05），and the above indicators in the study group were significantly lower than those in the control group（ P<0.05）. Conclusions:Synchronous brain bionic electrical stimulation in the hyperbaric oxygen chamber can improve the hemorheological indicators of the SD patients，reduce their blood hypercoagulability，and improve their hearing level and treatment effect. Hence，it is worth of clinical promotion.

Objective:To explore the curative effect of synchronous bionic electrical stimulation in hyperbaric oxygen chamber on patients with sudden deafness（SD）and the changes in hemorheological indicators.Methods:A total of 128 SD outpatients and inpatients treated at the Hyperbaric Oxygen Department of the Sixth Medical Center of the Chinese PLA General Hospital from June 2020 to June 2021 were randomly divided into study group（70 cases treated with synchronous brain bionic electrical stimulator in hyperbaric oxygen chamber）and control group（58 cases given conventional hyperbaric oxygen therapy），respectively. The curative effects of the two groups were evaluated，and the hemorheological indicators，such as the whole blood（high-shear and low-shear）viscosity，plasma viscosity，and red blood cell aggregation，were analyzed by the automatic biochemical analyzer before and after treatment.Results:After two courses of treatments，the overall effective rate of the study group（88.57%）was significantly higher than that of the control group（79.31%），and the difference was statistically significant（ P<0.05）. After treatment，the whole blood（high-shear and low-shear）viscosities，plasma viscosity，and erythrocyte aggregation indicators of the two groups were significantly decreased compared with those before treatment（ P<0.05），and the above indicators in the study group were significantly lower than those in the control group（ P<0.05）. Conclusions:Synchronous brain bionic electrical stimulation in the hyperbaric oxygen chamber can improve the hemorheological indicators of the SD patients，reduce their blood hypercoagulability，and improve their hearing level and treatment effect. Hence，it is worth of clinical promotion.

Objective To assess the efficacy of immunoprophylaxis against mother-to-infant transmission of hepatitis B virus (HBV) in a real-world setting since the implementation of charge-free hepatitis B immunoglobulin (HBIG) and hepatitis B vaccine in China. Methods The screening rate and the prevalence of hepatitis B surface antigen (HBsAg) in 61 790 puerperants, and the administration of combined immunoprophylaxis in infants of HBsAg-positive mothers in Rugao City of Jiangsu Province from July 1, 2011 to June 30, 2016 were retrospectively analyzed. HBV infection status of infants born to HBV infected mothers was followed up after 7 months of age. HBsAg-positive infants and their mothers were followed up again for HBV markers in April 2018. Fisher's exact test and Cochran-Armitage trend test were used to statistically analyze the differences between groups and various years, respectively. Results All 61 790 puerperants were screened for HBsAg and the prenatal screen rate was 98.6% (60 937/61 790) with an increasing trend over time (χ2trend=750.908, P<0.001). HBsAg-positive puerperants accounted for 5.5% (3 397/61 790) with a decreasing trend over time (χ2trend=32.667, P<0.001). In total, 778 offspring (399 boys and 379 girls) of 759 HBsAg-positive mothers were followed up at (13.7±6.9) months of age, among which 751 (96.5%) were administered and 25 (3.2%) were probably administered standard combined immunoprophylaxis after birth, and the rest two (0.3%) were not. Fourteen infants (1.8%) were HBsAg positive and all born to mothers with positive hepatitis B e antigen (HBeAg). None of the 538 infants born to HBeAg-negative mothers was HBsAg-positive. HBsAg-positive rate in infants born since 2013 was lower than those in 2011 and 2012 (χ2trend=13.352, P=0.000 3). Eleven HBsAg-positive mothers and their children were followed up again 4–5 years later in April 2018. HBV DNA levels of all mothers were within the range of (7.34–28.2)×107 IU/ml except one case of spontaneous HBeAg seroconversion. One out of the 11 infected children also had HBeAg seroconversion. Phylogenetic analysis of HBV S gene showed that the 11 pairs of mothers and children were all infected with HBV of genotype C. Conclusions The implementation of charge-free HBIG and hepatitis B vaccine for newborns achieves fruitful results in Rugao city as the mother-to-infant transmission of HBV in the real-world had been further reduced to a similar level reported in literature survey.

Objective To investigate the risk factors of surgical site infection (SSI) related complications after radical gastrectomy for gastric cancer in elderly patients.Methods The clinical data of 410 elderly patients with gastric cancer who underwent radical gastrectomy was retrospectively collected from 2009 to 2016.Univariate and multivariate analysis were performed to investigate the risk factors of SSI related complications,and the impact of SSI on short-term prognosis.Results SSI developed in 50 out of 410 elderly patients who underwent radical gastrectomy for gastric cancer,including 19 incisional infections and 31 organ lacuna infections.The corresponding incidence was 12.2％,4.6％ and 7.6％,respectively.By univariate analysis,age ＞ 75 (x2 =5.315,P =0.021),preoperative anemia (x2 =3.983,P =0.046),NRS 2002 ≥ 3 (x2 =4.785,P =0.029),diabetes (x2 =5.895,P =0.015),preoperative obstruction (x2 =5.250,P =0.022),undifferentiated carcinoma (x2 =4.448,P =0.035),cardiac carcinoma (x2 =5.265,P =0.022) and combined organs resection (x2 =4.165,P =0.041) were associated with SSI.Multivariate analysis showed that advanced age (OR =2.422,P =0.016),diabetes (OR =2.524,P =0.026),preoperative obstruction (OR =2.098,P =0.047) and high NRS 2002 score (OR =1.969,P =0.043) were independent risk factors for SSI.Conclusion The independent risk factors of SSI for elderly gastric cancer patients are advanced age,diabetes,preoperative obstruction and high NRS 2002 score.

Objective Study on the inhibitory effect of gallnut extract extract on MRSA β-lactamase. Methods Determination of inhibitory effect of gallnut extract on MRSA3002 by TTC method. β-lactamase was repeated by freezing and thawing method . Synergistic effect of gallnut extract and gentamicin was detected by TTC. Results The MIC and MBC of MRSA3002 by gallnut extract were 8mg/mL and 32mg/mL.Gallnut extract can reduce strains of β-lactamase activity,the MRSA300224h 1/2MIC after the effect of gallnut extract, beta lactam enzyme activity inhibition compared with the control group there were significant differences (P<0.01),compared with the positive control group, the difference was not significant. Synergistic effect of gallnut extract and gentamicin can significantly reduce the MIC of MRSA3002. Conclusion Gallnut extract can reduce β-lactamase activity recovery sensitivity of drug-resistant bacteria.

Objective:To determine the curative effect of Ginkgo Biloba extract on the back chapped skin in SD rats. Methods:Totally 60 SD rats were randomly divided into 6 groups with 10 rats in each. One group was the blank control group, while the other five groups with the same back chapped skin were respectively treated with different therapy regimen, namely one group was the model group without any treatment, one group was urea group treated with the commercially available urea, and the last three groups were re-spectively treated with Ginkgo Biloba extract at the dose of 5%, 1% and 0. 2%. The pathological tissue was observed after HE stai-ning, and the content of hydroxyproline ( Hyp) in skin was determined by alkaline hydrolysis. Results:The skin lesion and the score of pathological section were studied and the content of Hyp in the damaged skin was determined. The comprehensive score for the groups was 0, 13. 2, 7. 4, 11. 0, 5. 8 and 6. 6, respectively, and the content of Hyp was (4. 57 ± 0. 07), (2. 41 ± 0. 27), (3. 29 ± 0.57), (2.55 ±0.31), (3.24 ±0.26) and (3.37 ±0.24) μg·mg-1,respectively. There was significant difference in the curative effect between the Ginkgo Biloba extract groups at high dose (5%) and medium dose (1%) and the model group (P<0. 05), which showed no obvious difference when compared with the urea group (P>0. 05). Conclusion:The extract of Ginkgo Biloba has a certain effect on the back chapped skin in SD rats, which may contribute to the external preparations of Ginkgo Biloba extract.