Role of lactate dehydrogenase in diabetic neuropathic pain in mice: relationship with PGC-1α / 中华麻醉学杂志

Objective:To evaluate the role of lactate dehydrogenase in diabetic neuropathic pain (DNP) and the relationship with peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) in mice.Methods:SPF-grade healthy male C57BL/6J mice, aged 6-8 weeks, weighing 18-22 g, were used to establish diabetes mellitus model by intraperitoneal injection of streptozotocin (STZ) 120 mg/kg. Twenty-four mice with diabetes mellitus were divided into 2 groups ( n=12 each) using a random number table method: DNP group and DNP + oxamate group (OXA group). Another 12 SPF-grade healthy male C57BL/6J mice were selected as control group (C group). In OXA group, oxamate 750 mg/kg was intraperitoneally injected once a day for 28 consecutive days. The equal volume of normal saline was given instead in C group and DNP group. The mechanical paw withdrawal threshold (MWT), blood glucose and body weight were measured at 3 days before STZ injection and at 1, 2, 3 and 4 weeks after STZ injection (T 0-4). After the last behavioural test was completed, blood samples were collected from the posterior orbits of anesthetized mice for determination of serum lactate concentrations. The animals were then sacrificed and the tissues from the prefrontal cortex of the brain were taken for determination of lactate content, mitochondrial membrane potential (by the JC-1), content of reactive oxygen species (ROS) (using dihydroethidium probes), and level of histone lactylation and expression of PGC-1α (by Western blot). Results:Compared with C group, the MWT was significantly decreased at T 2-4, the serum lactate concentrations, contents of lactate and ROS and level of histone lactylation were increased, the mitochondrial membrane potential was decreased, and the expression of PGC-1α was down-regulated in DNP and OXA groups ( P<0.05). Compared with DNP group, no significant change was found in blood glucose and body weight ( P>0.05), the MWT was significantly increased at T 2-4, the serum lactate concentrations, contents of lactate and ROS and level of histone lactylation were decreased, the mitochondrial membrane potential was increased, and the expression of PGC-1α was up-regulated in OXA group ( P<0.05). Conclusions:Lactate dehydrogenase promotes the development of DNP, and the mechanism is related to promotion of increase in histone lactfication and down-regulation of PGC-1α expression in the prefrontal cortex of mice.

Shexiang Huangqi compound dripping pills-containing serum promotes proliferation and differentiation of bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Bone marrow mesenchymal stem cells have been widely used to treat neurological diseases.However,due to limitations of the blood-brain barrier,low survival rate and differentiation rate of stem cells at damaged sites,the therapeutic effect is limited. OBJECTIVE:To investigate the effects of Shexiang Huangqi compound dripping pills on proliferation,migration and astrocyte differentiation of bone marrow mesenchymal stem cells. METHODS:Male SD rats were treated with Shexiang Huangqi compound dripping pills for 5 days after continuous gavage.Blood was collected from the abdominal aorta and serum was separated for later use.The effect of 5%,10%and 20%drug-containing serum on the proliferation of bone marrow mesenchymal stem cells was detected by CCK-8 assay.The effect of 10%drug-containing serum on lateral migration of bone marrow mesenchymal stem cells was observed by scratch test.Bone marrow mesenchymal stem cells were cultured in Transwell cells.The effects of 10%drug-containing serum on longitudinal migration of bone marrow mesenchymal stem cells were observed by crystal violet staining and DAPI nuclear staining.Differentiation of bone marrow mesenchymal stem cells into astrocytes was observed by inducing solution with 10%drug-containing serum or co-culture with astrocytes. RESULTS AND CONCLUSION:(1)10%and 20%drug-containing serum promoted cell proliferation more significantly on days 2 and 3,and there was no statistical difference between the two concentrations.(2)At 30 and 48 hours,bone marrow mesenchymal stem cell migration in 10%drug-containing serum group was significantly higher than that in the control group.(3)The number of bone marrow mesenchymal stem cells filtered through Transwell cells in 10%drug-containing serum group was higher than that in the control group.(4)10%drug-containing serum might promote the differentiation of bone marrow mesenchymal stem cells to astrocytes,but the differentiation effect was weak,and astrocytes might further promote the differentiation of bone marrow mesenchymal stem cells into astrocytes induced by drug-containing serum.(5)The results exhibited that the 10%drug-containing serum could promote the proliferation and migration of bone marrow mesenchymal stem cells in vitro.Co-culture with astrocytes may promote the differentiation of bone marrow mesenchymal stem cells towards astrocytes.

Biallelic variants in RBM42 cause a multisystem disorder with neurological, facial, cardiac, and musculoskeletal involvement

Here, we report a previously unrecognized syndromic neurodevelopmental disorder associated with biallelic loss-of-function variants in the RBM42 gene. The patient is a 2-year-old female with severe central nervous system (CNS) abnormalities, hypotonia, hearing loss, congenital heart defects, and dysmorphic facial features. Familial whole-exome sequencing (WES) reveals that the patient has two compound heterozygous variants, c.304C>T (p.R102*) and c.1312G>A (p.A438T), in the RBM42 gene which encodes an integral component of splicing complex in the RNA-binding motif protein family. The p.A438T variant is in the RRM domain which impairs RBM42 protein stability in vivo. Additionally, p.A438T disrupts the interaction of RBM42 with hnRNP K, which is the causative gene for Au-Kline syndrome with overlapping disease characteristics seen in the index patient. The human R102* or A438T mutant protein failed to fully rescue the growth defects of RBM42 ortholog knockout ΔFgRbp1 in Fusarium while it was rescued by the wild-type (WT) human RBM42. A mouse model carrying Rbm42 compound heterozygous variants, c.280C>T (p.Q94*) and c.1306_1308delinsACA (p.A436T), demonstrated gross fetal developmental defects and most of the double mutant animals died by E13.5. RNA-seq data confirmed that Rbm42 was involved in neurological and myocardial functions with an essential role in alternative splicing (AS). Overall, we present clinical, genetic, and functional data to demonstrate that defects in RBM42 constitute the underlying etiology of a new neurodevelopmental disease which links the dysregulation of global AS to abnormal embryonic development.

Establishment of the normal reference values of left ventricular function parameters evaluated by CZT SPECT stress gated myocardial perfusion imaging in low-likelihood of stable coronary artery disease / 中华核医学与分子影像杂志

Objective:To establish the normal reference value of left ventricular function parameters by cadmium-zinc-tellurium (CZT) SPECT stress gated myocardial perfusion imaging (G-MPI) in low-likelihood of stable coronary artery disease (SCAD).Methods:From March 2022 to August 2022, 348 consecutive SCAD patients (146 males, 202 females, age (58±10) years) who underwent exercise or pharmacological stress G-MPI (CZT SPECT) in Beijing Anzhen Hospital, Capital Medical University were retrospectively recruited. Left ventricular end-diastolic volume (EDV), end-systolic volume (ESV), and left ventricular ejection fraction (LVEF) were acquired using quantitative gated SPECT (QGS) analysis. EDV and ESV were corrected by body surface area (BSA) to obtain EDV index (EDVI) and ESV index (ESVI), respectively. Independent-sample t test, one-way analysis of variance and Mann-Whitney U test were used for data analysis. The influences of EDV, ESV, EDVI, ESVI and LVEF were analyzed by multiple regressions for linear models. Results:There were 314 patients with low-likelihood of SCAD (128 males, 186 females, age (58±10) years) and 34 normal controls (18 males, 16 females, age (55±10) years). There were no significant differences of basic clinical characteristics and left ventricular function parameters in different genders between 2 groups ( z values: from -1.74 to -0.02, t values: from -1.16 to 1.17, all P>0.05). Using the 95% CI as the cut-off value for left ventricular function parameters in patients with a low-likelihood of SCAD, the upper limits of EDV, ESV, EDVI and ESVI in females and males were 84 and 111 ml, 30 and 44 ml, 47 and 54 ml/m 2, 17 and 21 ml/m 2, respectively, and the lower limit of LVEF in females and males were 58% and 55%, respectively. In the low-likelihood of SCAD group, the EDV ((58±13) vs (77±17) ml) and ESV ((16±7) vs (26±9) ml) of females were smaller than those of males ( t values: 10.65, 10.35, both P<0.001), while LVEF of females was higher than that of males ((72±7)% vs (67±6)%; t=-6.23, P<0.001). However, there were no significant differences in left ventricular function parameters among different age groups with the same gender ( F values: 0.12-2.19, all P>0.05). Based on multiple regression for linear models, the primary predictors of EDV, ESV and LVEF were gender and weight ( β values: from -0.380 to 0.358, all P<0.05). Conclusions:Normal reference values of left ventricular function parameters are established by CZT SPECT stress G-MPI in low-likelihood of SCAD patients. Left ventricular EDV and ESV of females are smaller than those of males, while LVEF of females is higher than that of males. The influence of gender on left ventricular function parameters should be considered in clinical practice.

Effect of metformin preconditioning on AMPK/PINK1 signaling pathway during myocardial ischemia-reperfusion injury in diabetic rats / 中华麻醉学杂志

Objective:To evaluate the effect of metformin preconditioning on adenosine monophosphate-activated protein kinase(AMPK)/PTEN-induced putative protein kinase(PINK1) signaling pathway during ischemia-reperfusion (I/R) injury in diabetic rats.Methods:Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 6 weeks, weighing 120-160 g, were divided into 3 groups ( n=12 each) by the random number table method: diabetic sham operation group (DS group), diabetic myocardial I/R group (DI/R group) and diabetic myocardial I/R+ metformin preconditioning group(DI/R+ Met group). After 4 weeks of feeding a high-fat and high-glucose diet, the model of type 2 diabetes mellitus was induced by a single intraperitoneal injection of 1% streptozotocin 40 mg/kg. The myocardial I/R injury was induced by blocking the anterior descending branch of the left coronary artery for 30 min followed by 120-min reperfusion in anesthetized animals. In DI/R+ Met group, metformin 200 mg/kg was given by intragastric gavage once a day within 1 week before myocardial ischemia. Blood samples from the femoral vein were collected at 120 min of reperfusion for determination of the serum creatine kinase isoenzymes (CK-MB) and cardiac troponin I (cTnI) concentrations by enzyme-linked immunosorbent assay. Then the rats were sacrificed and myocardial tissues were obtained for examination of the pathological changes(by HE staining) and for determination of the percentage of myocardial infarct size (by the double staining of Ewan blue and TTC) and expression of myocardial autophagy-related protein Beclin-1, PTEN-induced putative kinase 1 (PINK1), phosphorylated 5′-adenosine monophosphate-activating protein kinase (p-AMPK), and ratio of microtubule-associated protein 1 light chain 3Ⅱ/Ⅰ (LC3Ⅱ/Ⅰ) (by Western blot). Results:Compared with DS group, the percentage of myocardial infarct size and serum CK-MB and cTnI concentrations were significantly increased, the expression of Beclin-1, p-AMPK and PINK1 in myocardial tissues was up-regulated, the ratio of LC3II/I was increased( P<0.05), and the pathological changes were aggravated in DI/R group and DI/R+ Met group. Compared with DI/R group, the percentage of myocardial infarct size and serum CK-MB and cTnI concentrations were significantly decreased, the expression of Beclin-1, p-AMPK and PINK1 in myocardial tissues was up-regulated, the ratio of LC3Ⅱ/Ⅰ was increased ( P<0.05), and the pathological changes were significantly reduced in DI/R+ Met group. Conclusions:The mechanism by which metformin preconditioning reduces myocardial I/R injury is related to activation of AMPK/PINK1 signaling pathway and up-regulation of mitochondrial autophagy in diabetic rats.

Effect of resveratrol on ferroptosis in cardiomyocytes of mice with diabetic cardiomyopathy / 中华麻醉学杂志

Objective:To evaluate the effect of resveratrol on ferropotosis in cardiomyocytes of mice with diabetic cardiomyopathy.Methods:Thirty healthy adult male C57BL/6 mice, aged 8 weeks, weighing 22-26 g, were divided into 3 groups ( n=10 each) using a random number table method: control group (group C), diabetic cardiomyopathy group (group DCM) and resveratrol group (group RSV). Freshly prepared streptozotocin (STZ) 40 mg·kg -1·d -1 was intraperitoneally injected for 5 consecutive days to develop the model of type 1 diabetes mellitus. After the model was successfully developed, resveratrol 25 mg·kg -1·d -1 was intragastrically given for 12 consecutive weeks in group RSV, while the equal volume of dimethyl sulfoxide was given instead in group C and group DCM. Echocardiography was performed to examine the cardiac structure and function at the end of the 12th week. Then mice were sacrificed, and myocardial tissue specimens were harvested for microscopic examination of the pathological changes of myocardial tissues (by Hematologist-Eosin staining) and mitochondrial morphology of myocardial cells (with a transmission electron microscope) and for determination of the contents of iron, malondialdehyde (MDA) and glutathione (GSH) (by colorimetry) and expression of glutathione peroxidase 4 (GPX4) (by Western blot). Results:Compared with group C, the left ventricular end-diastolic diameter and left ventricular end-systolic diameter were significantly increased, the left ventricular ejection fraction and left ventricular fractional shortening were decreased, the contents of iron and MDA were increased, the content of GSH was decreased, and the expression of GPX4 was down-regulated in group DCM ( P<0.05). Compared with group DCM, the left ventricular end-diastolic diameter and left ventricular end-systolic diameter were significantly decreased, the left ventricular fractional shortening and ejection fraction were increased, the contents of iron and MDA were decreased, the content of GSH was increased, the expression of GPX4 was up-regulated ( P<0.05), and the pathological changes of myocardial tissues and changes in mitochondrial morphology of myocardial cells were significantly attenuated in group RSV. Conclusions:The mechanism by which resveratrol attenuates myocardial injury and further improves cardiac dysfunction is related to inhibition of ferroptosis in cardiomyocytes of mice with diabetic cardiomyopathy.

elationship between preoperative fasting plasma glucose and postoperative pulmonary complications after lung resection in type 2 diabetic patients / 中国胸心血管外科临床杂志

@#Objective     To explore the relationship between preoperative fasting plasma glucose (FPG) and postoperative pulmonary complications (PPCs) in type 2 diabetic patients undergoing elective thoracoscopic lung resection, and provide a reference for prediction and prevention of PPCs in the clinic. Methods     A retrospective analysis was performed on the type 2 diabetic patients who underwent elective thoracoscopic lung resection for the first time in our hospital from January 2017 to March 2021. According to the level of FPG one day before the operation, the patients were divided into three groups: a hypoglycemia group (<6.1 mmol/L), a medium level blood glucose group (≥6.1 mmol/L and <8.0 mmol/L) and a high blood glucose group (≥8.0 mmol/L). Besides, the patients were divided into a PPCs group and a non-PPCs group according to whether PPCs occurred. The risk factors for PPCs were analyzed by logistic regression analysis, and the predictive value of preoperative FPG level on PPCs was estimated by the area under the receiver operating characteristic curve (AUC). Results     A total of 130 patients were included, including 75 (57.7%) males and 55 (42.3%) females with an average age of 63.5±9.0 years. Logistic regression analysis showed that compared to non-PPCs patients, the level of preoperative FPG (P=0.023) and smoking history ratio (P=0.036) were higher and the operation time was longer (P=0.004) in the PPCs patients. High FPG level on preoperative day 1 and longer operation time were associated with PPCs risk. Besides, the preoperative FPG of 6.79 mmol/L was the threshold value to predict the occurrence of PPCs [AUC=0.653, 95%CI (0.559, 0.747), P=0.003]. Conclusion     There is a certain correlation between preoperative FPG level and postoperative PPCs, which may be used as an index to predict the occurrence of PPCs.

Application of fluorescence in situ hybridization technique to verify the clonalities of non-clonal cytogenetic abnormalities identified in Myelodysplastic syndrome / 中华医学遗传学杂志

OBJECTIVE@#To assess the value of fluorescence in situ hybridization (FISH) technique for the verification of the clonalities of non-clonal cytogenetic abnormalities (n-CCA) identified by conventional chromosome banding analysis (CBA) in patients with Myelodysplastic syndrome (MDS).@*METHODS@#Clinical data and results of karyotyping and FISH assays for 91 patients of MDS with n-CCA identified by CBA were retrospectively analyzed. In total 94 non-clonal +8, 5q-, -7/7q- or 20q- were detected by CBA, among which 43 (45.7%) were verified to be clonal abnormalities by FISH.@*RESULTS@#The detection rates for +8, 5q-, -7/7q- and 20q- by FISH were 47.6% (30/63), 25% (2/8), 41.7% (5/12), 40% (2/5) and 66.7% (4/6), respectively, with the positive cells accounting for 4% to 90% of all counted cells, with a median value of 7%. The 91 patients were divided into three groups including ≥ 20, 10 ~< 20 and < 10 based on the numbers of metaphase cells in CBA, and the detection rates by FISH for the three groups were 43.7% (31/71), 33.3% (3/9) and 63.6% (7/11), respectively, which showed no statistically difference (P > 0.05). Continuous CBA and FISH surveys were conducted for 26 patients who received supportive treatment, and the results revealed that 91.7% (11/12) of FISH-verified positive abnormalities had persisted, whereas 92.9% (13/14) of the n-CCA verified as negative by FISH was transient.@*CONCLUSION@#Nearly half of the CBA identified n-CCA have been verified as clonal aberrations by FISH, and the FISH detection rate showed no correlation with the number of metaphase cells. FISH test is strongly recommended for verifying the clonalities of n-CCA detected by CBA, and continuous cytogenetic survey of the patients with MDS is necessary.

Analysis of the characteristics of primary acute myeloid leukemia with 11q23/KMT2A rearrangements in ninety patients / 中华医学遗传学杂志

OBJECTIVE@#To investigate the clinical and prognostic characteristics of primary acute myeloid leukemia (AML) with 11q23/KMT2A rearrangements.@*METHODS@#Clinical data of 90 patients with primary AML and 11q23/KMT2A rearrangements were analyzed retrospectively.@*RESULTS@#By karyotyping analysis, 80 of the 90 patients had translocations involving 11q23/KMT2A, with t(9;11)(p22;q23), t(6;11)(q27;q23), t(10;11)(p12;q23) and t(11;19)(q23;p13) being the most common ones, while 10 cases were found to have non-translocation abnormalities. The overall complete remission (CR) rate was 75.6%, and patients with t(6;11) had lower CR rate compared with non-t(6;11) patients (47.1% vs. 82.2%, P = 0.005). After a median follow-up of 24.5 months, the patients receiving allo-hematopoietic stem cell transplantation (allo-HSCT) had significantly higher 3-year overall survival (OS) (80.3% vs. 16.6%, P < 0.001) and 3-year event-free survival (EFS) (73.5% vs. 16.3%, P < 0.001) compared with non-transplant patients. Patients with t(6;11) had the lowest 3-year OS (11.8% vs. 56.0%, P < 0.001) and 3-year EFS (5.9% vs. 53.8%, P < 0.001) compared with other type of abnormalities. No significant difference was noted in the survival between patients with t(9;11) and non-t(9;11) regardless whether they had received HSCT.@*CONCLUSION@#The clinical characteristics of primary AML with 11q23/KMT2A rearrangements are heterogeneous. Patients did not receive HSCT had poorer survival, particularly with the presence of t(6;11). Allo-HSCT could significantly improve the survival of such patients.

Case report: Prenatal diagnosis of Ectrodactyly-Ectodermal dysplasia-Cleft syndrome (EEC) in a fetus with cleft lip and polycystic kidney.

Ectrodactyly-ectodermal dysplasia-cleft (EEC) syndrome is an autosomal dominant disorder characterized by ectrodactyly, ectodermal dysplasia, and orofacial clefting. Reduced penetrance is manifested in these core features and additional under-recognized features, especially in prenatal cases. Here, we present a fetus with EEC syndrome at 22 weeks gestation, in which the cleft lip and palate and the right polycystic kidney are shown by prenatal ultrasound. A de novo missense mutation of R304W in the TP63 gene is confirmed by whole-exome sequencing associated with EEC syndrome. We further investigate the reported TP63-related prenatal cases and provide a more complete picture of the prenatal phenotypic spectrum about EEC. It illustrates the potential severity of genitourinary anomalies in TP63-related disorders and highlights the need to counsel for the possibility of EEC syndrome, given the occurrence of genitourinary anomalies with orofacial cleft or limb deformities.

Post-treatment With Irisin Attenuates Acute Kidney Injury in Sepsis Mice Through Anti-Ferroptosis via the SIRT1/Nrf2 Pathway.

Kidney is one of the most vulnerable organs in sepsis, resulting in sepsis-associated acute kidney injury (SA-AKI), which brings about not only morbidity but also mortality of sepsis. Ferroptosis is a new kind of death type of cells elicited by iron-dependent lipid peroxidation, which participates in pathogenesis of sepsis. The aim of this study was to verify the occurrence of ferroptosis in the SA-AKI pathogenesis and demonstrate that post-treatment with irisin could restrain ferroptosis and alleviate SA-AKI via activating the SIRT1/Nrf2 signaling pathway. We established a SA-AKI model by cecal ligation and puncture (CLP) operation and an in vitro model in LPS-induced HK2 cells, respectively. Our result exhibited that irisin inhibited the level of ferroptosis and ameliorated kidney injury in CLP mice, as evidenced by reducing the ROS production, iron content, and MDA level and increasing the GSH level, as well as the alteration of ferroptosis-related protein (GPX4 and ACSL4) expressions in renal, which was consistent with the ferroptosis inhibitor ferrostatin-1 (Fer-1). Additionally, we consistently observed that irisin inhibited ROS accumulation, iron production, and ameliorated mitochondrial dysfunction in LPS-stimulated HK-2 cells. Furthermore, our result also revealed that irisin could activate SIRT1/Nrf2 signaling pathways both in vivo and vitro. However, the beneficial effects of irisin were weakened by EX527 (an inhibitor of SIRT1) in vivo and by SIRT1 siRNA in vitro. In conclusion, irisin could protect against SA-AKI through ferroptotic resistance via activating the SIRT1/Nrf2 signaling pathway.

Effects of different anesthesia methods on immune function in patients with oral squamous cell carcinoma / 中华麻醉学杂志

Objective:To evaluate the effect of different anesthesia methods on the immune function in the patients with oral squamous cell carcinoma.Methods:Forty patients of both sexes, aged 31-64 yr, with body mass index of 19-23 kg/m 2, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, undergoing elective radical resection of oral squamous cell carcinoma and repair of the defect with free flap, were enrolled and randomized to receive either combined intravenous-inhalational anesthesia (VICA group, n=20) or total intravenous anesthesia (TIVA group, n=20) using a random number table method.In group VICA, anesthesia was induced with intravenous propofol 1.5-2.0 mg/kg, remifentanil 1-2 μg/kg, and cisatracurium 0.2 mg/kg, sevoflurane was continuously inhaled to maintain MAC at 1.3, sevoflurane inhalation was stopped at 1 h before the end of surgery, sevoflurane was replaced with propofol, propofol 4-6 mg·kg -1·h -1 was continuously infused until the end of operation, and dexmedetomidine 0.4 μg·kg -1·h -1, remifentanil 0.2-0.3 μg·kg -1·min -1 and cisatracurium 0.1 mg·kg -1·h -1 were intravenously infused at the same time to maintain anesthesia.In group TIVA, anesthesia induction was the same as those previously described in group VICA, and anesthesia was maintained with intravenous dexmedetomidine 0.4 μg·kg -1·h -1, propofol 4-6 mg·kg -1·h -1, remifentanil 0.2-0.3 μg·kg -1·min -1 and cisatracurium 0.1 mg·kg -1·h -1.Venous blood samples were taken at 30 min before anaesthesia induction (T 0), 3 h after anaesthesia (T 1), at the end of operation (T 2), and at 6, 24 and 48 h after operation (T 3-5) for determination of the serum concentrations of immunoglobulins (IgA, IgM, IgG), interleukins (IL-2, IL-6, IL-10, sIL-2Rα) and soluble interleukin-2 receptor alpha (sIL-2Rα) by enzyme-linked immunosorbent assay. Results:Compared with the baseline at T 0, the concentrations of serum sIL-2Rα at T 1-5, IL-2 at T 1-4 and IL-10 at T 1 were significantly decreased, the concentrations of serum IL-6 at T 1-5 and IL-10 at T 2-4 were increased, and the concentrations of serum IgA and IgM at T 1-5 were decreased in two groups, and the concentrations of serum IgG at T 1-5 in TIVA group and at T 1, 2 and T 4, 5 in VICA group were significantly decreased ( P<0.05).Compared with group TIVA, the concentrations of serum sIL-2Rα at T 2, 5, IL-6 at T 4, 5 and IL-10 at T 3, IgA at T 4 and IgG at T 3 were significantly increased, and the concentrations of serum IL-2 at T 1-5 and IgA at T 5 were decreased in group VICA ( P<0.05). Conclusions:Both general anesthesia methods have significant inhibitory effects on intraoperative and postoperative cellular immune function and humoral immune function in the patients with oral squamous cell carcinoma, and combined intravenous-inhalational anesthesia produces higher inhibitory effects on cellular immune function than total intravenous anesthesia.

Role of Shh/Gli1 signaling pathway in sleep deprivation-induced cognitive impairment in young mice / 中华麻醉学杂志

Objective:To evaluate the role of sonic hedgehog (Shh)/glioma-associated oncogene homolog 1 (Gli1) signaling pathway in sleep deprivation-induced cognitive impairment in young mice.Methods:Forty-eight SPF healthy male C57BL/6 mice, aged 4 weeks, weighing 14-16 g, were divided into 3 groups ( n=16 each) by the random number table method: control group (C group), sleep deprivation group (SD group) and Shh agonist SAG group (SD+ SAG group). Multi-platform water environment method was used to prepare the sleep deprivation model in mice, and the sleep deprivation was 20 h a day for 10 consecutive days.In SD+ SAG group, SAG 10 mg/kg was intraperitoneally injected at 5 min before each sleep deprivation, while the equal volume of normal saline was intraperitoneally injected in group C and group SD.The mice underwent novel object recognition and Y-maze tests at 24 h after development of the model.Mice were sacrificed after the behavioral testing, and the hippocampi were isolated for determination of the density of dendritic spines in hippocampal CA1 region (by Golgi staining), expression of Gli1 and brain-derived neurotrophic factor (BDNF) in hippocampal tissues (by Western blot), and expression of Gli1 and BDNF mRNA in hippocampal tissues (by quantitative real-time polymerase chain reaction). Results:Compared with group C, the preference index in novel object recognition and Y-maze tests and density of dendritic spines in CA1 region were significantly decreased, and the expression of Gli1 and BDNF protein and mRNA in hippocampus was down-regulated in group SD ( P<0.05). Compared with group SD, the preference index in novel object recognition and Y-maze tests and density of dendritic spines in CA1 region were significantly increased, and the expression of Gli1 and BDNF protein and mRNA in hippocampus was up-regulated in group SD+ SAG ( P<0.05). Conclusions:Inhibition of Shh/Gli1 signaling pathway and reduction of plasticity of dendritic spines of hippocampal neurons are involved in sleep deprivation-induced cognitive impairment in young mice.

Role of Yes-associated protein 1 in acute lung injury in septic mice: the relationship with ferroptosis / 中华麻醉学杂志

Objective:To evaluate the role of Yes-associated protein 1 (YAP1) in acute lung injury (ALI) and the relationship with ferroptosis in septic mice.Methods:Twenty-four male wild-type mice and 24 YAP1 conditional knockout mice, aged 9-10 weeks, weighing 22-25 g, were divided into 2 groups ( n=12 each) using a random number table method: wild-type sham operation group (WT+ Sham group) and wild-type sepsis-induced ALI group (WT+ ALI group); YAP1 conditional knockout sham operation group (CKO+ Sham group) and YAP1 conditional knockout sepsis-induced ALI group (CKO+ ALI group). The sepsis-induced ALI model was developed by cecal ligation and perforation (CLP) in anesthetized animals.The bronchoalveolar lavage fluid (BALF) was collected at 24 h after CLP to determine the protein concentration (by bicinchoninic acid method) and concentrations of interleukin-1beta (IL-1β) and tumor necrosis factor-α (TNF-α) (by enzyme-linked immunosorbent assay). Mice were then sacrificed, and the lung tissues were obtained for examination of ultrastructure (using a transmission electron microscope) and for determination of wet/dry lung weight ratio (W/D ratio), contents of Fe 2+ , malondialdehyde (MDA) and glutathione (GSH) (by colorimetric assay), and expression of YAP1, glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4) and solute carrier family 7 member 11 (SLC7A11) (by Western blot). Results:Compared with WT+ Sham group, the concentrations of protein in BALF, IL-1β and TNF-α were significantly increased, W/D ratio and contents of Fe 2+ and MDA were increased, GSH contents were decreased, the expression of GPX4 and SLC7A11 was down-regulated, ACSL4 expression was up-regulated ( P<0.05), alveolar epithelial cells showed characteristic changes of ferroptosis with mitochondrial shrinkage and decreased mitochondrial cristae in WT+ ALI group.Compared with WT+ CLP and CKO+ Sham groups, the concentrations of protein in BALF, IL-1β and TNF-α were significantly increased, W/D ratio and contents of Fe 2+ and MDA were increased, GSH contents were decreased, the expression of GPX4 and SLC7A11 was down-regulated, ACSL4 expression was up-regulated ( P<0.05), and the mitochondria in alveolar epithelial cells in lung tissues shrank obviously, and the mitochondrial cristae were reduced or even disappeared in CKO+ CLP group ( P<0.05). Conclusions:YAP1 is involved in the endogenous protective mechanism against ALI, which is related to inhibition of ferroptosis in septic mice.

Morphological alterations study in the Golgi apparatus of skin fibroblasts in spinocerebellar ataxia type 3 patients / 中华神经科杂志

Objective:To identify the morphological alterations in the Golgi apparatus of skin fibroblasts in spinocerebellar ataxia type 3 (SCA3) patients.Methods:In this study, 3 SCA3 patients and 3 healthy volunteers were obtained in the First Affiliated Hospital of Zhengzhou University from 2016 to 2020. The cytosine, adenine, and guanine repeats of 3 SCA3 patients were 14/76, 20/80 and 21/82, respectively. Tissue mass culture was used to amplify skin fibroblasts derived from SCA3 patients and healthy volunteers. Cell viability and apoptosis were detected using cell counting kit-8 assay and flow cytometry, respectively. Western blotting and immunofluorescence assay were used to detect the protein expression of ataxin-3, Golgi reassembly stacking protein 2 (GORASP2), and Golgi matrix protein 130 (GM130) in the skin fibroblasts. The morphology of the Golgi apparatus in skin fibroblasts was detected using transmission electron microscopy.Results:Tissue culture of skin fibroblasts of both SCA3 patients and healthy volunteers was successfully established. The patient-derived dermal fibroblasts expressing mutant ataxin-3 protein exhibited reduced cell viability ( t=5.06,P=0.007), increased apoptosis ( t=3.77, P=0.020), fragmentation of the Golgi apparatus, increased expression of GM130 ( t=5.23, P=0.006), and decreased expression of GORASP2 ( t=4.35, P=0.012). Transmission electron microscopy revealed that the Golgi apparatus was disorganized in skin fibroblasts. Conclusion:Fragmentation of the Golgi apparatus occurs in the skin fibroblasts of SCA3 patients, and abnormal morphology and structure of the Golgi apparatus may be involved in the pathogenesis of SCA3.

Relationship between Nrf2 and ferroptosis during lung ischemia-reperfusion in rats / 中华麻醉学杂志

Objective:To evaluate the relationship between nuclear factor E2-related factor 2 (Nrf2) and ferroptosis during lung ischemia-reperfusion (I/R) in rats.Methods:Fifty-four healthy male Sprague-Dawley rats, weighing 220-250 g, were divided into 3 groups ( n=18 each) using a random number table method: sham operation group (Sham group), lung I/R group (IR group), and lung I/R+ Nrf2 agonist sulforaphane group (IR+ SFP group). Lung I/R model was developed by clamping the left pulmonary hilum for 60 min followed by 120 min of reperfusion.In IR+ SFP group, SFP 500 μg/kg was intraperitoneally injected at 3 days before lung ischemia once a day for 3 consecutive days, and the model was developed at 2 h after the end of administration.The rats were sacrificed at the end of reperfusion, and the bronchoalveolar lavage fluid (BALF) was collected to determine the protein concentration (using bicinchoninic acid method), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) concentrations (by enzyme-linked immunosorbent assay). The animals were then sacrificed, and lung tissue specimens were harvested for microscopic examination of the pathological changes (with a transmission electron microscope) and for determination of wet/dry weight (W/D) ratio, contents of iron, malondialdehyde (MDA) and glutathione (GSH) (by chemical colorimetric) and expression of nuclear Nrf2, glutathione peroxidase 4 (GPX4) and acyl-CoA synthetase long-chain family member 4 (ACSL4) (by Western blot). Results:Compared with Sham group, the concentrations of protein, IL-6 and TNF-α in BALF, W/D ratio, and contents of Fe 2+ and MDA were significantly increased, GSH content was decreased, GPX4 expression was down-regulated, the expression of nuclear Nrf2and ACSL4 was up-regulated ( P<0.05), and the mitochondrial morphology of type Ⅱalveolar epithelial cells showed the characteristic changes of ferroptosis, including the presence of smaller mitochondria and reduced cristae in IR group.Compared with IR group, the concentrations of protein, IL-6 and TNF-α in BALF, W/D ratio, and contents of Fe 2+ and MDA were significantly decreased, GSH content was increased, the expression of nuclear Nrf2 and GPX4 expression was up-regulated, ACSL4 expression was down-regulated ( P<0.05), and the pathological changes of lung tissues were significantly attenuated in IR+ SFP group. Conclusions:Nrf2 can inhibit ferroptosis during lung I/R and is involved in the endogenous protective mechanism in rats.

Effects of propofol on neuronal activity in medial prefrontal cortex during social behavior in sleep-deprived rats / 中华麻醉学杂志

Objective:To evaluate the effect of propofol on neuronal activity in medial prefrontal cortex (mPFC) during social behavior in sleep-deprived rats.Methods:Sixty SPF healthy male Sprague-Dawley rats, aged 8 weeks, weighing 200-250 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group Con), chronic sleep deprivation plus natural sleep group (group CSD+ NS), and chronic sleep deprivation plus propofol group (CSD+ Pro). Sleep deprivation model was developed by the modified multiple platform method, and the rats were placed in the sleep-deprivation tank for 20 h a day (14: 00-10: 00) and allowed to sleep naturally for 4 h (10: 00-14: 00) a day for 28 consecutive days.Propofol 40 mg/kg was intraperitoneally injected after sleep deprivation for 28 consecutive days in group CSD+ Pro, while the equal volume of 10% fat emulsion was given instead in group C and group CSD+ NS.Electroencephalographic recordings in cerebral cortical regions were performed on the days 1st, 14th and 28th after sleep deprivation.The apoptotic neurons in mPFC were detected using TUNEL method after the end of sleep deprivation, and the apoptosis index was calculated.A three chamber sociability test was used to detect the social behavior of rats, and local field potential signals in mPFC were collected. Results:Compared with group Con, the percentage of rapid eye movement sleep was significantly increased, the sniffing time preference coefficients in the 2 stages were reduced, the percentage of the β waves and θ waves-band power in mPFC during the social sniffing process was decreased, and the apoptosis index of neurons in mPFC was increased in group CSD+ NS ( P<0.05). Compared with group CSD+ NS, the percentage of rapid eye movement sleep was significantly increased, the sniffing time preference coefficient in the 2 stages was increased, and the percentage of β waves and θ waves-band power in mPFC during the social sniffing process was increased, and the apoptosis index of neurons in mPFC was decreased in group CSD+ Pro ( P<0.05). Conclusions:Propofol inhibits the apoptosis in neurons in mPFC and increases β and θ waves in the mPFC during social interaction after sleep deprivation in sleep-deprived rats, which is helpful in improving sleep deprivation-induced social disorder.

UBXN3B Restricts Viral Pathogenesis by Maintaining Hematopoietic Homeostasis

Hematopoiesis is finely regulated to enable timely production of the right numbers and types of mature immune cells to maintain tissue homeostasis. Dysregulated hematopoiesis may compromise antiviral immunity and/or exacerbate immunopathogenesis. Herein, we report an essential role of UBXN3B in maintenance of hematopoietic homeostasis and restriction of immunopathogenesis during respiratory viral infection. Ubxn3b deficient (Ubxn3b-/-) mice are highly vulnerable to SARS-CoV-2 and influenza A infection, characterized by more severe lung immunopathology, lower virus-specific IgG, significantly fewer B cells, but more myeloid cells than Ubxn3b+/+ littermates. This aberrant immune compartmentalization is recapitulated in uninfected Ubxn3b-/- mice. Mechanistically, UBXN3B controls precursor B-I (pre-BI) transition to pre-BII and subsequent proliferation in a cell-intrinsic manner, by maintaining BLNK protein stability and pre-BCR signaling. These results reveal an essential role of UBXN3B for the early stage of B cell development.

Changes in blood-cerebrospinal fluid barrier in postoperative delirium rats / 中华麻醉学杂志

Objective:To evaluate the changes in the blood-cerebrospinal fluid barrier in postoperative delirium rats.Methods:One hundred and forty-seven healthy female Sprague-Dawley rats, aged 3 months, weighing 240-300 g, were divided into 3 groups ( n=49 each) using a random number table method: control group (group C), anesthesia group (group A) and postoperative delirium group (group P). Group C received no treatment.Group A received 2-h anesthesia with 1.4% isoflurane.Group S underwent an exploratory laparotomy under 1.4% isoflurane anesthesia.The behaviors of rats in each group were tested at 24 h before surgery and 6, 9 and 24 h after surgery using buried food test, open field test and Y maze test.Sodium fluorescence was injected through the tail vein at 6, 9 and 24 h after surgery.Then the rats were sacrificed, the choroid plexus (CP) was obtained, and cerebrospinal fluid (CSF) of bilateral cerebral ventricles was collected, and the expression of ZO-1, occludin, claudin1, E-cadherin and VE-cadherin in CP was detected using Western blot.FITC-dextran 10, 40 and 70 kDa was injected through the tail vein at 6 h after surgery, and then CSF was collected for determination of the concentrations of NaFI, 10, 40 and 70 kDa fluorescein isothiocyanate labeled dextran (FITC-dextran) in CSF by fluorescence spectrophotometry.CP was obtained to observe the morphology of choroid plexus epithelial cells (CPECs) of bilateral cerebral ventricles with a transmission electron microscope. Results:Compared with group C and group A, the latency to eat food in buried food test was significantly prolonged, the time of staying at the central region was shortened, the percentage of the number of entries into novel arm and percentage of time of staying at novel arm in Y maze test were decreased, the freezing time in open field test was shortened, the expression of ZO-1, occludin and claudin1 in CP was down-regulated, the concentrations of NaFI and 10 kDa and 40 kDa FITC-dextranin CSF were increased ( P<0.05 or 0.01), the CPECs arranged at random and loose, the microvilli of CPECs were absent, the tight junction was blurred, and the gap became wider in group P. Conclusion:The occurrence of postoperative delirium is related to the change in blood-cerebrospinal fluid barrier.

Role of miR-146a in hippocampal inflammatory responses in postoperative cognitive dysfunction in mice / 中华麻醉学杂志

Objective:To evaluate the role of miR-146a in hippocampal inflammatory responses in postoperative cognitive dysfunction (POCD) in mice.Methods:One hundred and sixty clean-grade male C57BL/6 mice, aged 12-16 weeks, weighing 22-28 g, were divided into 5 groups ( n=32 each) using a random number table method: control group (group C), group POCD, miR-146a agomir group (group Ag), miR-146a antagomir group (group At) and negative control group (group NC). The mice were subjected to an intramedullary fixation for tibial fracture under 1.5% isoflurane anesthesia to establish POCD model.At 2 days before operation, miR-146a agomir 0.5 nmol (0.1 nmol/μl) was injected into bilateral hippocampi in group Ag, miR-146a antagomir 2.5 nmol (0.5 nmol/μl) was injected in group At, miR-146a negative control solution 2.5 nmol (0.5 nmol/μl) was given in group NC, and the animals in group C did not receive any treatment.At 1 day before operation and at 1, 3 and 7 days after operation, open-field test was performed to evaluate spontaneous motor activity, and contextual fear conditioning test was performed to evaluate cognitive ability 15 min later.At 1 and 3 days after operation, the animals were sacrificed and hippocampi was removed for determination of expression of CD11b (a marker for activation of microglia) in hippocampal CA1 region by immunofluorescence staining.At 6, 12 and 24 h after operation, the expression of miR-146a was detected by quantitative real-time polymerase chain reaction, the expression of interleukin-1 receptor-associated kinase 1 (IRAK1), tumor necrosis factor receptor-associated factor 6 (TRAF6), nuclear factor kappa B p65 (NF-κB p65) and tumor necrosis factor-alpha (TNF-α) was determined by Western blot and interleukin-1beta (IL-1β) and IL-6 contents were determined by enzyme-linked immunosorbent assay. Results:There was no significant difference in the total exploring distance in the open-field test or percentage of freezing time in tone-fear conditioning test at each time point among the five groups( P>0.05). Compared with group C, the percentage of freezing time in the contextual fear conditioning test was significantly decreased at 1, 3 and 7 days after operation, the expression of CD11b at 1 and 3 days after surgery and expression of miR-146a, IRAK1, TRAF6, NF-κB p65 and TNF-α were up-regulated and the contents of IL-1 β and IL-6 were increased at 6, 12 and 24 h after operation in group POCD ( P<0.05). Compared with group NC, the percentage of freezing time in the contextual fear conditioning test was significantly increased at 1, 3 and 7 days after operation, and the expression of CD11b was down-regulated at 1 and 3 days after surgery, and the expression of miR-146a, IRAK1, TRAF6, NF-κB p65 and TNF-α was up-regulated and IL-1β and IL-6 contents were decreased at 6, 12 and 24 h after operation in group Ag, and the percentage of freezing time in the contextual fear conditioning test was decreased at 1, 3 and 7 days after operation, the expression of CD11b at 1 and 3 days after surgery was up-regulated, the expression of miR-146a was down-regulated and IRAK1, TRAF6, NF-κB p65 expression was up-regulated at 6, 12 and 24 h after operation, TNF-α expression was up-regulated and IL-1β and IL-6 contents were increased at 12 and 24 h after operation in group At ( P<0.05). Conclusion:miR-146a is involved in the process of hippocampal inflammatory responses, and the mechanism may be related to the inhibition of IRAK1-TRAF6-NF-κB signaling pathway in mice.