Objective:To evaluate the role of ERBB2 interacting protein (Erbin)in liver tissues in blood coagulation of septic mice.Methods:Thirty SPF healthy male C57BL/6 mice and 30 Erbin knockout mice, aged 8-10 weeks, weighing 20-30 g, were divided into wild-type+ sham operation group (WT+ Sham group), wild-type+ sepsis group (WT+ SEP group), Erbin gene knockout+ sham operation group (EKO+ Sham group) and Erbin gene knockout+ sepsis group (EKO+ SEP group) by a random number table method, with 15 animals in each group. The mouse sepsis model was prepared by the cecal ligation and perforation method in anesthetized animals. Eye blood samples were collected at 24 h after surgery and liver tissues were obtained for microscopic examination of histopathological changes (by HE staining) which were scored and for determination of plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities (by colorimetry), expression of Erbin and tissue factor (TF) (by Western blot), expression of tissue plasminogen activator (t-PA) and fibrinogen (Fib)mRNA (by quantitative polymerase chain reaction), concentrations of PT, APTT, thrombin time (TT) and Fib (by automatic coagulation analyzer), and plasma TF and interleukin-6 (IL-6) concentrations (by enzyme-linked immunosorbent assay).Results:Compared with WT+ Sham group, the lung injury score was significantly increased, the expression of TF, t-PA mRNA and FGA mRNA was up-regulated, PT, APTT and TT were prolonged, the plasma Fib concentration was increased, and the activities of ALT and AST and concentrations of TF and IL-6 in plasma were increased in WT+ SEP group ( P<0.05). Compared with WT+ SEP group, the lung injury score was significantly increased, the expression of TF, t-PA mRNA and FGA mRNA was up-regulated, PT, APTT and TT were prolonged, the plasma Fib concentration was increased, and the activities of ALT and AST and concentrations of TF and IL-6 in plasma were increased in EKO+ SEP group ( P<0.05). Conclusions:Erbin in liver tissues exerts an endogenous protective effect on blood coagulation by inhibiting the up-regulation of TF expression in septic mice.

Background and purpose:The overexpression of carnitine palmitoyl transferase 1A(CPT1A)is related to the poor prognosis of breast cancer,and it can promote the utilization of fatty acids by mitochondria and maximize triphosphate(ATP)production.However,the role of CPT1A in breast cancer metastasis is still unclear.This study aimed to explore the mechanism that mitochondrial dysfunction and CPT1A/extracellular signal-regulated kinase(ERK)signaling pathway in breast cancer jointly regulate the malignant behavior of breast cancer.Methods:The lentivirus system and shRNA tools were used to overexpress or knock down CPT1A in human breast cancer cell lines MDA-MB-231 and MCF7,and the cells were divided into NC group,CPT1A group and shCPT1A group.The invasion ability of cells was detected by transwell assay,and the protein expressions of peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α),ERK1/2 and CPT1A were analyzed by Western blot.The mitochondrial morphology of MDA-MB-231 and MCF7 cell lines was analyzed using mitochondrial red staining,and mitochondrial respiratory capacity was analyzed by oxygen consumption rate.Results:Compared with cells expressing control vector,overexpression of CPT1A resulted in enhanced invasion abilities of MCF7 cells and MDA-MB-231 cells(P＜0.05),while knockdown of shCPT1A resulted in decreased invasion abilities of cells(P＜0.05).Compared with NC group,the length of mitochondrial branches in MDA-MB-231 and MCF7 cells in CPT1A group was significantly shorter(P＜0.05),and the expressions of ERK1/2 and PGC-1α increased significantly(P＜0.05).In shCPT1A group,the length of mitochondrial branches in MDA-MB-231 and MCF7 cells increased significantly(P＜0.05),and the expressions of ERK1/2 and PGC-1α decreased significantly(P＜0.05).In addition,compared with NC group,the cellular basis,maximum respiratory capacity and ATP production of MDA-MB-231 in CPT1A group increased significantly(P＜0.05),while the cellular basis,maximum respiratory capacity and ATP production of MDA-MB-231 in shCPT1A group decreased significantly(P＜0.05).Conclusion:ERK1/2-PGC-1α activated by CPT1A Signaling pathway plays a key role in mitochondrial division mediated breast cancer cell metastasis.

[摘 要] 目的：探讨四个半LIM结构域2（FHL2）蛋白对胶质瘤细胞增殖的影响及其分子机制。方法：利用TCGA和CGGA数据库分析胶质瘤组织中FHL2 mRNA表达水平与患者预后的关系。通过WB法检测人胶质瘤组织标本及人胶质瘤细胞U87、T98G、U251、SNB19、GSC23、A172、LN229、G267和星形胶质细胞NHA中的FHL2蛋白表达水平。利用慢病毒载体构建稳定敲低FHL2的U87细胞和过表达FHL2的SNB19细胞，即U87-shGFP、U87-shFHL2-1#、U87-shFHL2-4#和SNB19-3flag、SNB19-3flag-FHL2组。通过CCK-8法、克隆形成实验检测敲低和过表达FHL2对细胞增殖的影响，免疫共沉淀（Co-IP）和液相色谱-串联质谱（LC/MS）法筛选FHL2在胶质瘤细胞中的相互作用蛋白，并用Co-IP和免疫荧光法验证它们的结合作用和共定位情况。使用酶标仪检测敲低和过表达FHL2细胞内乳酸产量和乳酸脱氢酶（LDH）活性的变化，WB法分析FHL2、LDHA及p-LDHA在正常脑组织和胶质瘤组织中的蛋白表达差异及其相互关系。在过表达 FHL2的SNB19细胞中使用LDHA的小分子抑制剂AT-101，通过CCK-8实验和酶标仪比色法验证FHL2在胶质瘤乳酸代谢中的作用，验证AT-101在胶质瘤中潜在的治疗效果。结果：Co-IP和LC/MS检测发现，FHL2与LDHA在胶质瘤细胞中存在相互作用。FHL2过表达可提高LDHA活性和乳酸生成（均P < 0.001），进而促进胶质瘤细胞增殖（P < 0.001）。相反，敲低FHL2会降低LDHA活性和乳酸产量（P < 0.001或P < 0.05）并抑制细胞增殖（P < 0.001）。AT101能抑制LDHA活性，并显著抑制FHL2促进胶质瘤细胞的增殖，同时恢复磷酸化LDHA（Y10）水平（P<0.01或P < 0.001）。结论：FHL2与LDHA蛋白相互作用，FHL2通过激活p-LDHA（Y10）的表达促进LDHA活性和乳酸产生，进而促进胶质瘤细胞的增殖，靶向这种相互作用可能成为治疗胶质瘤的潜在策略。

AIM:To investigate the impact of celastrol intervention on excitatory amino acid transporter 2(EAAT2)and its neuroprotective role in subarachnoid hemorrhage(SAH).METHODS:Western blot analysis was uti-lized to assess the EAAT2 expression level within 72 h after SAH,while glutamate concentration in cortical brain tissues was measured.Computational simulation was employed to explore the binding of celastrol with EAAT2.Seventy SD rats were randomly assigned to sham,model,model+GT949(an EAAT2 agonist),model+dihydrokainic acid(DHK;an EAAT2 inhibitor),and model+celastrol groups.Glutamate concentration in cortical brain tissues was quantified,and brain edema was assessed by dry-wet weight method.Western blot analysis was conducted to evaluate the expression of EAAT2,aquaporin 4 and apoptosis-related proteins(Bax,Bcl-2,caspase-3 and caspase-9),and TUNEL staining was employed to assess the apoptotic cell count in each group.RESULTS:(1)EAAT2 level decreased while glutamate con-centration increased.(2)Celastrol was found to directly bind to EAAT2,enhancing EAAT2 expression and reducing glu-tamate concentration after SAH.(3)Celastrol demonstrated the ability to inhibit brain edema after SAH.(4)Celastrol was effective in reducing neuronal apoptosis after SAH.CONCLUSION:Celastrol has the potential to up-regulate EAAT2 expression,lower glutamate level,mitigate brain edema,and decrease neuronal apoptosis after SAH.

Objective:To explore the relationship between glycosylated hemoglobin A 1c/high-density lipoprotein cholesterol ratio (HbA 1c/HDL-C) and urinary albumin-creatinine ratio (UACR) in Chinese adults. Methods:In this cross-sectional study, the clinical data of 43 820 community residents (age>40 years) from the Risk Evaluation of Cancers in Chinese Diabetic Individuals (REACTION study; March-December 2012) across eight centers (Liaoning, Guangdong, Shanghai, Gansu, Guangxi, Henan, Hubei, and Sichuan) in China were collected and analyzed. Participants were divided into three groups based on UACR levels:<10 mg/g, 10-30 mg/g, and >30 mg/g. The HbA 1c/HDL-C ratio was divided into four groups according to quartile division of the subjects: 1st quartile (Q1<3.79), 2nd quartile (3.79≤Q2<4.59), 3rd quartile (4.59≤Q3≤5.66), and 4th quartile (Q4>5.66). Multivariate ordinal logistic regression model was used to analyze the relationship between HbA 1c/HDL-C and UACR. Receiver operating characteristic (ROC) analysis was used to explore the predictive value of HbA 1c/HDL-C to UACR. Results:The 43 820 subjects included 13 452 (30.70%) male and 30 378 (69.30%) female patients, with an average age of (58.00±0.05) years. According to results of one-way analysis of variance analysis, the HbA 1c/HDL-C ratio was significantly associated with the risk of increased UACR ( F=495.73, P<0.001). After adjusting for clinically relevant confounding variables in logistic regression model, compared with participants with the lowest HbA 1c/HDL-C ratio (Q1), women with the highest HbA 1c/HDL-C ratio (Q4) had a 1.483-fold (95% CI 1.376-1.598, P<0.001) and men had a 1.161-fold (95% CI 1.019-1.323, P<0.001) increased risk of UACR. The ROC curve analysis showed that the area under the curve of HbA 1c/HDL-C for predicting increased UACR was 0.623 (95% CI 0.597-0.606), with a sensitivity of 60.18% and a specificity of 54.91%. The HbA 1c/HDL-C ratio showed the highest predictive value of all glycemic and lipidemic parameters. In individuals with well-controlled blood glucose (HbA 1c<6.5%) or lipid levels (HDL-C≥1.0 mmol/L), the HbA 1c/HDL-C ratio was still independently associated with the risk of increased UACR after adjusting for confounding variables [ OR(95% CI) of quartile 4: 1.563 (1.210-2.019, P=0.001) in participants with HbA 1c<6.5% and 1.822 (1.687-1.968, P<0.001) in participants with HDL-C≥1.0 mmol/L]. Conclusion:As a novel compound indicator for evaluating glucose homeostasis and dyslipidemia, the HbA 1c/HDL-C ratio was independently associated with increased UACR in the general population aged>40 years in China, which was superior to both glycemic and lipid parameters alone.

Objective:To analyze the clinical features, treatment, and outcomes of fetal/neonatal atrial flutter (AFL) at the onset of the perinatal period to improve the management of this condition.Methods:This retrospective study analyzed the clinical data, treatment, and follow-up results of fetal/neonatal AFL cases transferred to Shanghai Children's Medical Center from November 2013 to August 2021. Clinical characteristics, cardioversion procedures, and outcomes were summarized. Descriptive method was used for statistical analysis.Results:A total of 21 fetuses/neonates presenting with AFL in the perinatal period were involved in this study, including 17 males and four females. Ten of them were born at full term, and 11 were preterms. All of the patients were delivered by cesarean section at 32 to 41 gestational weeks [ (36.6±1.9) weeks] with a birth weight of 2 130 to 4 450g [ (3 059±528) g]. Increased fetal heart rate was all detected after 32 weeks of gestation, and three of them were diagnosed with AFL by fetal echocardiography before being born. The heart rate remained elevated in all cases after birth. All were diagnosed as AFL based on an electrocardiogram on the day of birth, which showed a 2 to 6 over one ratio of atrioventricular conduction. Among the six cases of cardiac insufficiency and low blood pressure complicated by dyspnea and cyanosis, the symptoms were relieved in four cases after mask oxygenation and two cases after ventilation. Among the 21 cases, one was converted spontaneously to normal sinus rhythm and the other 20 recovered after medication or electrical cardioversion. Seven cases were initially treated by drug conversion with a success rate of 5/7 and hospitalized for 23 d (13-25 d). There was one with cardiac insufficiency before treatment and three newly developed cardiac insufficiency during treatment among the seven cases. Thirteen cases were offered electrical cardioversion initially, and the success rate of cardioversion was 12/13. There were five cases of cardiac insufficiency before treatment, while no new cases of cardiac insufficiency was reported during treatment. The duration of hospitalization was 11 d (9-14 d). Apart from one case, the rest 20 infants were followed up from one month to eight years old, and no recurrence was reported.Conclusions:For fetal/neonatal AFL with the onset during the perinatal period, the symptoms mainly manifest in late pregnancy. Its diagnosis depends on fetal echocardiography before birth or electrocardiogram after birth, and electrical cardioversion is a fast and effective measure. While the prognosis of perinatal-onset AFL is generally good.

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.

【Objective】 To detect and analyze the infection status of HBsAg non-reactive /HBV DNA reactive blood donors by individual donor-NAT (ID-NAT) and chemiluminescence technology, and to explore the feasibility and potential risks of reentry. 【Methods】 The blood screening results of blood donors in Wuhu from January 2018 to October 2021 were queried by blood station information management software. The blood donation information of all HBsAg non-reactive /HBV DNA reactive blood donors was collected and then recalled by telephone. After informed consent, samples were taken for HBV DNA nucleic acid single test, enzyme-linked immunoassay for HBsAg, chemiluminescence assay for HBV seromarkers(including HBsAg, anti-HBs, HBeAg, anti-HBe and anti-HBc), and alanine aminotransferase (ALT) test. All the results were statistically analyzed. 【Results】 From January 2018 to October 2021, there were 142 051 donations, and the positive rate of sole HBV DNA was 0.06% (91/142 051), and 33 people (37 person-times) were successfully followed up. The yield rates of HBsAg, anti-HBs and anti-HBc were 6.06% (2/33), 39.39% (13/33) and 96.97% (32/33), respectively; None HBeAg was yielded. After two times of ID-NAT, 8 patients remained non-reactive to both systems, with a negative conversion rate of 24.24% (8/33). Meanwhile, 25 patients were at least once reactive to ID-NAT, and 23 of them were occult HBV infection with serologically reactivity. There were 2(6.25%) patients with HBsAg positive conversion and HBV DNA persistent reactivity, which were window period infection. One person was confirmed as false reactivity (no HBV infection) as he remained unreactive to both repeated ID-NAT and serological tests. 【Conclusion】 Chemiluminescence assay is more sensitive than ELISA in detecting HBV serum markers, which is beneficial to early detection of HBV samples in window period. The yielding rate of anti-HBc among HBsAg non-reactive/HBV DNA reactive blood donors detected by blood screening in this region is very high, and most of them are occulting infection, so the ID-NAT should be no less than 2 times in the reentry strategy.

OBJECTIVE: To investigate the effect of acetylcorynoline (Ace) for promoting functional recovery of injured spinal cord in rats and explore the underlying mechanism. METHODS: Rat models of spinal cord injury (SCI) were treated with intraperitoneal injection of different concentrations of Ace, with the sham-operated rats as the control group. After the treatment, the changes in motor function of the rats and the area of spinal cord injury were assessed with BBB score and HE staining, and the changes in pro-inflammatory cytokine levels and microglial activation were determined using PCR, ELISA and immunofluorescence staining. In a lipopolysaccharide (LPS)-treated BV2 cell model, the effects of different concentrations of Ace or DMSO on microglial activation and inflammatory cytokine production were observed. Network pharmacology analysis was performed to predict the target protein and signaling mechanism that mediated the inhibitory effect of Ace on microglia activation, and AutoDock software was used for molecular docking between Ace and the target protein. A signaling pathway blocker (Osimertinib) was used to verify the signaling mechanism in rat models of SCI and LPS-treated BV2 cell model. RESULTS: In rat models of SCI, Ace treatment significantly increased the BBB score, reduced the area of spinal cord injury, and lowered the number of activated microglia cells and the levels of pro-inflammatory cytokines (P < 0.05). The cell experiments showed that Ace treatment significantly lower the level of cell activation and the production of inflammatory cytokines in LPS-treated BV2 cells (P < 0.05). Network pharmacology analysis suggested that EGFR was the main target of Ace, and they bound to each other via hydrogen bonds as shown by molecular docking. Western blotting confirmed that Ace inhibited the activation of the EGFR/MAPK signaling pathway in injured mouse spinal cord tissue and in LPS-treated BV2 cells, and its inhibitory effect was comparable to that of Osimertinib. CONCLUSION In rat models of SCI, treatment with Ace can inhibit microglia-mediated inflammatory response by regulating the EGFR/MAPK pathway, thus promoting tissue repair and motor function recovery.
Mice ; Animals ; Rats ; Recovery of Function ; Lipopolysaccharides ; Microglia ; Molecular Docking Simulation ; Spinal Cord Injuries ; Signal Transduction ; Cytokines ; ErbB Receptors

Objective:To investigate the clinical and radiological outcomes of ultrasound guided injection of platelet-rich plasma (PRP) in the treatment of intratendinous rotator cuff tear.Methods:A retrospective study was conducted to analyze the clinical data of 43 patients (46 shoulders) who had been treated for intratendinous partial-thickness rotator cuff tear by ultrasound guided injection of PRP consecutively from July 2021 to March 2022 at Department of Sports Medicine, Peking University Third Hospital. There were 23 males and 20 females, with an age of (47.8±13.5) years and a course of disease of 6 (4, 18) months, involving 22 left shoulders and 24 right shoulders. The visual analog scale (VAS) pain score, the University of California at Los Angeles (UCLA) rating scale, and the shoulder index of the American Shoulder and Elbow Surgeons (ASES) were determined before injection and at the last follow-up. The changes in tear size were also evaluated by magnetic resonance imaging (MRI) before PRP injection and 3 to 5 months after PRP injection.Results:The 43 patients were followed up for 15 (12, 17) months after treatment. Of this cohort, 7 shoulders (15.2%, 7/46) were recovered to complete normal and very satisfied with the injection effects while 19 shoulders(41.3%, 19/46) satisfied with the effects after injection, yielding an overall satisfaction rate of 56.5% (26/46). At the last follow-up, the VAS score [3.0 (2.0, 4.0) points], ASES score [80.0 (65.0, 88.8) points], and UCLA score [29.0 (20.0, 32.0) points] were significantly improved compared with those before injection [5.5 (4.0, 8.0) points, 55.0 (39.2, 65.0) points, and 16.0 (12.0, 20.3) points] ( P < 0.05). MRI evaluation showed the tear volume was significantly reduced after PRP injection [46.1 (20.9, 77.5) mm 3 before injection versus 28.2 (12.5, 63.6) mm 3 after injection] ( P<0.05), and a >50% tear volume diminution was observed in 13 shoulders (34.2%,13/38). There were no complications during or after injection. Conclusion:As the ultrasound guided injection of PRP into intratendinous lesions is effective and safe for patients with intratendinous partial-thickness rotator cuff tear, it can be an alternative treatment for the patients or professional athletes who are unwilling to undergo surgery.