Objective To investigate the effects and mechanism of Ferrostatin-1(Fer-1),a ferroptosis inhibitor,on myocardial ischemia-reperfusion injury(MIRI).Methods Rat H9c2 cardiomyocytes were ran-domly divided into five groups:Control group,H/R medium group,H/R medium+Fer-1 group,H/R medium+Nec-1 group,and H/R medium+emricasan group.Cell morphology was observed using electron mi-croscopy.Cell proliferation activity was assessed via CCK-8 assay and lactate dehydrogenase(LDH)release.I-ron ion levels were measured using an iron detection kit.Reactive oxygen species(ROS)and mitochondrial su-peroxide levels were detected by flow cytometry and MitoSOXTM fluorescence staining,respectively.Western blot was employed to analyze the expression of glutathione peroxidase 4(GPX4),acyl-CoA synthetase long-chain family member 4(ACSL4),nicotinamide adenine dinucleotide phosphate oxidase(NOX1),and cycloox-ygenase 2(COX2).Results Compared to the Control group,the H/R medium group exhibited significantly increased cytotoxicity(LDH levels)and reduced cell viability,with statistically significant differences(P<0.05).Treatment with Fer-1,Nec-1,or emricasan in the H/R medium group increased cell adherence,reduced vacuolization,enhanced cell viability,and decreased cytotoxicity(LDH relative releasing rate)compared to the H/R medium group.Intracellular ferrous iron and total iron levels were elevated in the H/R medium group compared to the Control group,with statistically significant differences(P<0.05),while Fer-1 treatment sig-nificantly reduced these levels(P<0.05).ROS levels were higher in the H/R medium group than in the Con-trol group,and Fer-1 treatment attenuated this increase(P<0.05).Western blot analysis revealed elevated ACSL4,NOX1,and COX2 levels,alongside reduced GPX4 levels,in the H/R medium group compared to the Control group,with statistically significant differences(P<0.05).Fer-1 treatment reversed these trends,de-creasing ACSL4,NOX1,and COX2 levels while increasing GPX4 expression,with statistically significant differences(P<0.05).Conclusion Ferroptosis plays a critical role in MIRI.Fer-1 mitigates oxidative stress injury and alleviates MIRI by inhibiting ferroptosis.

Objective:To analyze the clinical data and genetic characteristics of a child with CLN1 neuronal ceroid lipofuscinosis in conjunct with hereditary hyperferritinemia cataract syndrome (HHCS).Methods:A child who was admitted to the PICU of the First Affiliated Hospital of Zhengzhou University in November 2020 was selected as the study subject. Clinical data of the child was collected. Genetic testing was carried out for the child, and the result was analyzed in the light of literature review to explore the clinical and genetic characteristics to facilitate early identification.Results:The patient, a 3-year-old male, had mainly presented with visual impairment, progressive cognitive and motor regression, and epilepsy. Cranial magnetic resonance imaging revealed deepened sulci in bilateral cerebral hemispheres, and delayed myelination. The activity of palmitoyl protein thioesterase was low (8.4 nmol/g/min, reference range: 132.2 ~ 301.4 nmol/g/min), whilst serum ferritin was increased (2 417.70 ng/mL, reference range: 30 ~ 400 ng/mL). Fundoscopy has revealed retinal pigment degeneration. Whole exome sequencing revealed that he has harbored c. 280A>C and c. 124-124+ 3delG compound heterozygous variants of the PPT1 gene, which were respectively inherited from his father and mother. Neither variant has been reported previously. The child has also harbored a heterozygous c. -160A>G variant of the FTL gene, which was inherited from his father. Based on the clinical phenotype and results of genetic testing, the child was diagnosed as CLN1 and HHCS. Conclusion:The compound heterozygous variants of the PPT1 gene probably underlay the disorders in this child. For children with CLN1 and rapidly progressing visual impairment, ophthalmological examination should be recommended, and detailed family history should be taken For those suspected for HHCS, genetic testing should be performed to confirm the diagnosis.

Objective:To further improve the understanding of paroxysmal nocturnal hemoglobinuria (PNH), we retrospectively analyzed and summarized the clinical characteristics, treatment status, and survival status of patients with PNH in Zhejiang Province.Methods:This study included 289 patients with PNH who visited 20 hospitals in Zhejiang Province. Their clinical characteristics, comorbidity, laboratory test results, and medications were analyzed and summarized.Results:Among the 289 patients with PNH, 148 males and 141 females, with a median onset age of 45 (16-87) years and a peak onset age of 20-49 years (57.8% ). The median lactic dehydrogenase (LDH) level was 1 142 (604-1 925) U/L. Classified by type, 70.9% (166/234) were classical, 24.4% (57/234) were PNH/bone marrow failure (BMF), and 4.7% (11/234) were subclinical. The main clinical manifestations included fatigue or weakness (80.8%, 235/289), dizziness (73.4%, 212/289), darkened urine color (66.2%, 179/272), and jaundice (46.2%, 126/270). Common comorbidities were hemoglobinuria (58.7% ), renal dysfunction (17.6% ), and thrombosis (15.0% ). Moreover, 82.3% of the patients received glucocorticoid therapy, 70.9% required blood transfusion, 30.7% used immunosuppressive agents, 13.8% received anticoagulant therapy, and 6.3% received allogeneic hematopoietic stem cell transplantation. The 10-year overall survival (OS) rate was 84.4% (95% CI 78.0% -91.3% ) . Conclusion:Patients with PNH are more common in young and middle-aged people, with a similar incidence rate between men and women. Common clinical manifestations include fatigue, hemoglobinuria, jaundice, renal dysfunction, and recurrent thrombosis. The 10-year OS of this group is similar to reports from other centers in China.

Objective To investigate the effect of perineal nerve electroacupuncture combined with tolterodine in the treatment of female overactive bladder(OAB).Methods A total of 150 female OAB patients treated in Huzhou Hospital of Traditional Chinese Medicine,Zhejiang Chinese Medical University from April 2020 to May 2021 were selected and divided into control group and study group according to random number table method,with 75 cases in each group.The control group was treated with tolterodine,and the study group was treated with pudendal nerve electroacupuncture combined with tolterodine.Clinical symptoms,urinary symptoms,and urodynamic indicators[maximum cystometric capacity(MCC),maximum urethral closure pressure(Pura clos max),maximum detrusor pressure(Pdet max),and maximum urine flow rate(Qmax)],bladder function indicators(initial bladder sensation volume,bladder volume during strong micturition desire),nerve growth factor(NGF)mediated transient receptor potential(TRP)pathway indicators[NGF,NGF/urine creatinine(UCr),TRPV1,TRPV4]and clinical efficacy of two groups were compared.Results After treatment,the number of night urination and urgent urination in study group were significantly less than those in control group,and the average volume of urination was significantly more than that in control group(P＜0.05).The overactive bladder symptom scores and visual analogue scale scores of study group were significantly lower than those of control group(P＜0.05).The MCC and Pura·clos·max in study group were significantly higher than those in control group,while Pdet max and Qmax were significantly lower than those in control group(P＜0.05).The initial bladder sensation volume and bladder volume during strong micturition desire in study group were higher than those in control group(P＜0.05).NGF,NGF/UCr,positive expression rates of TRPV1 and TRPV4 in study group were lower than those in control group(P＜0.05).The total effective rate was significantly higher in study group than in control group,(x2=5.374,P=0.020).There was no significant difference in the incidence of total adverse reactions between two groups(x2=0.362,P=0.547).Conclusion Perineal nerve electroacupuncture combined with tolterodine can significantly improve bladder function and inhibit clinical symptoms in female OAB patients,possibly by inhibiting bladder sensitivity caused by NGF-mediated increase of TRP channel proteins TRPV1 and TRPV4.

Objective To investigate the predictive value of white blood cell-to-hematocrit ratio (WBCHR) for in-hospital major adverse cardiovascular events (MACE) after reperfusion therapy in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods A case-control study was conducted to retrospectively select 319 patients with first-time diagnosis of STEMI who underwent percutaneous coronary intervention (PCI). Patients were divided into MACE group (69 cases) and non-MACE group (250 cases) based on the occurrence of MACE during hospitalization. Clinical data, including general information, laboratory test indicators, echocardiography, and coronary angiography results, were compared between the two groups. Univariate and multivariate Logistic regression analyses were performed to explore the risk factors for in-hospital MACE after reperfusion therapy in STEMI patients. Receiver operating characteristic (ROC) curve analysis was conducted to evaluate the predictive value of WBCHR for in-hospital MACE after reperfusion therapy in STEMI patients. Results The levels of fasting blood glucose, uric acid, creatinine, white blood cell count, neutrophil count, high-sensitivity C-reactive protein (hs-CRP), D-dimer, and WBCHR were significantly higher in the MACE group than in the non-MACE group, while red blood cell count, hemoglobin, hematocrit, and left ventricular ejection fraction were lower (P < 0.05). No significant differences were observed in age, gender, smoking history, hypertension history, diabetes history, door-to-wire (D2W) time, total cholesterol, triglycerides, and the number of diseased vessels between the two groups (P>0.05). Multivariate Logistic regression analysis revealed that elevated uric acid levels (OR=1.005; 95%CI, 1.002 to 1.009; P=0.004), decreased hemoglobin concentration (OR=0.964; 95%CI, 0.941 to 0.988; P=0.003), increased hs-CRP levels (OR=1.032; 95%CI, 1.009 to 1.056; P=0.007), and increased WBCHR (OR=1.455; 95%CI, 1.295 to 1.635; P < 0.001) were independent risk factors for in-hospital MACE after reperfusion therapy in STEMI patients. ROC curve analysis showed that the area under the curve of WBCHR for predicting in-hospital MACE after reperfusion therapy in STEMI patients was 0.855 (95%CI, 0.796 to 0.914, P < 0.001), with a sensitivity of 63.8% and a specificity of 99.6%. Conclusion WBCHR is an independent influencing factors of in-hospital MACE after reperfusion therapy in STEMI patients and has a high predictive value for in-hospital MACE in these patients.

Different fragments of SARS-CoV-2 nucleocapsid (N) protein were expressed and purified, and a fluorescence immunochromatography method for detection of SARS-CoV-2 total antibody was established. The effect of different protein fragments on the performance of the method was evaluated. The N protein sequence was analyzed by bioinformatics technology, expressed in prokaryotic cell and purified by metal ion affinity chromatography column. Different N protein fragments were prepared for comparison. EDC reaction was used to label fluorescence microsphere on the synthesized antigen to construct sandwich fluorescence chromatography antibody detection assay, and the performance was systemically evaluated. Among the 4 prepared N protein fragments, the full-length N protein (N419) was selected as the optimized coating antigen, N412 with 0.5 mol/L NaCl was used as the optimal combination; deleting 91-120 amino acids from the N-terminal of N412 reduced non-specific signal by 87.5%. the linear range of detection was 0.312-80 U/L, the limit of detection was 0.165 U/L, and the accuracy was more than 95%. A fluorescence immunochromatographic detection method for analysis of SARS-CoV-2 total antibody was established by pairing N protein fragments. The detection result achieved 98% concordance with the commercially available Guangzhou Wanfu test strip, which is expected to be used as a supplementary approach for detection of SARS-CoV-2. The assay could also provide experimental reference for improving the performance of COVID-19 antibody detection reagents.
Antibodies, Viral ; COVID-19 ; Chromatography, Affinity ; Fluorescent Antibody Technique ; Humans ; Microspheres ; SARS-CoV-2 ; Sensitivity and Specificity

Objective:To investigate the relationship between the E2 and E4 alleles of apolipoprotein E (apoE) gene and myocardial infarction (MI) in type 2 diabetes Mellitus (T2DM) patients, and to explore the relationship between apoE polymorphism and blood lipid metabolism.Methods:This case control study was conducted from August 2016 to March 2020 in China-Japan Friendship Hospital, 3 459 inpatients with T2DM were included including 3 044 patients without MI (T2DM group) and 415 patients with MI (T2DM+MI group). Real time fluorescent quantitative PCR was used to detect apoE polymorphism. Automatic biochemical analyzer was used to detect lipid levels. Logistic regression analyses were performed to determine the association of apoE with risk of MI in patients with T2DM.Results:(1) The frequency of E4 allele in T2DM+MI group (12.29%, 102/830) was significantly higher than in T2DM group (9.13%,556/6 088), while the frequency of E2 allele in T2DM+MI group (7.35%,61/830) was significantly lower than that in T2DM group (8.21%,500/6 088), P=0.012. Logistic regression analyses showed that E4 allele carrier (E3/E4+E4/E4) faced a higher risk for MI in T2DM patients ( OR=1.48, 95% CI 1.14-1.92, P=0.003), while E2 allele carrier(E2/E3+E2/E2)did not face a higher risk of MI in T2DM patients ( OR=0.88, P=0.642). (2) The levels of apoE polymorphism and blood lipid: The levels of TC, LDL-C and apoB increased in the order of E4 allele, wild type and E2 allele ( P<0.05). The levels of HDL-C, apoA1 and apoE decreased in the order of E4 allele, Wild type and E2 allele ( P<0.05). Conclusion:The E4 allele is a risk factor for MI in T2DM patients, and apoE polymorphism can affect blood lipid level in this patent cohort.

Objective To evaluate the effect and mechanism of tribulus terrestris in kidney calcium oxalate stone in rats.Methods Sisty healthy male Wistar rats were divided into control group,model group,low-dose prevention group,high-dose prevention group,low-dose treatment group,high-dose treatment group by random number table method,each group of 10 rats.The control group was had free access to water,the model group was given 1％ ethylene glycol had free access to water,and the low-dose prevention group was given 1％ ethylene glycol had free access to water while was added in Tribulus terrestris extract 250 mg/(kg · d) treated by gavage,the high-dose prevention group was given 1％ ethylene glycol had free access to water while was added in Tribulus terrestris extract 500 mg/(kg · d) treated by gavage.The low-dose treatment group received 1％ ethylene glycol had free access to water while was added in tribulus terrestris extract 250 mg/(kg · d) treated by gavage on the 15th day.The high-dose treatment group received 1％ ethylene glycol had free access to water while was added in tribulus terrestris extract 500 mg/(kg · d) treated by gavage on the 15th day.At the end of the experiment on the 28th day,the body weight,blood urea nitrogen,creatinine,P,Ca2+,Mg2+ content,24 h urine volume,24 h urine Ca2+,Mg2 +,P,oxalic acid secretion,renal tissue homogenate detection of superoxide dismutase (SOD),glutathione peroxidase (GPX),malondialdehyde content of each group were detected.The crystal deposition and pathological changes of calcium oxalate were observed under the microscope.Measurement data were expressed as mean ± standard deviation (Mean ± SD),multiple comparisons between groups were performed using LSD-t test,and comparison between groups was performed by one-way anova analysis of variance.Results Compared with the control group,the rats in the model group had blood urea nitrogen (28.93 ±3.37) mmol/L,creatinine (162.31 ± 34.08) μmol/L,24 h uric acid (132.67 ± 27.96) μmol,Ca2+ (18.24 ± 1.94) μmol/24 h,the content of malondialdehyde (191.04 ± 20.56) nmol/mgprot in kidney homogenate was significantly increased,blood Ca2+ (2.01 ±0.28) mmol/L,renal GPX (479.82 ± 136.97) μmol/gprot,SOD(1.54 ±0.21) U/mgprot decreased significantly,these differences were statistically significant (P ＜ 0.05).Renal histopathology showed low-dose and high-dose prevention and treatment groups kidney calcium oxalate crystals and kidney damage were less than the model group.Conclusion The extract of Tribulus terrestris has a beneficial effect against calcium oxalate urinary stone formation induced by ethylene glycol.

Objective To investigate the clinical efficacy of free flap transplantation in repairing the Gustilo type ⅢB and ⅢC fractures of tibia and fibula combined with soft tissue defects.Methods A retrospective case series study was conducted on the clinical data of 46 patients who had Gustilo type ⅢB and ⅢC fractures of tibia and fibula with soft tissue defects admitted from June 2013 to January 2017.There were 34 males and 12 females,aged 1-67 years (mean,39 years).The wound defect areas ranged from 6 cm × 20 cm to 7 cm × 38 cm.According to the Gustilo fracture classification,there were 31 cases of type ⅢB and 15 cases of type ⅢC.According to the AO fracture typing,there were five cases of type A,23 type B,and 18 type C.All patients were repaired with free flap transplantation,among which 40 patients were treated with anterolateral thigh flap and six with latissimus dorsi flap.The areas of anterolateral thigh flap ranged from 6 cm × 13 cm to 14 cm ×32 cm,and those of the latissimus dorsi flap from 6 cm × 22 cm to 7 cm × 40 cm.Efficacy was evaluated by flap survival rate,complications,fracture healing time,lower limb function scoring system (LEFS),and skin flap function.Results All limbs were salvaged successfully.One case of total flap necrosis and eight cases of postoperative crisis occurred.After active exploration and treatment,three cases were seen distal local necrosis,and the total survival rate was 91％.Infection at the donor site was found in two cases.The complication incidence rate was 4％.All patients were followed up for 7-42 months,with an average of 19 months.The fracture healing time averaged 43.5 weeks,and the LEFS score averaged 54 points.According to the seven indexes of flap function,the results were excellent in 1 1 cases,good in 29 cases,fair in four cases,and poor in two cases,with an excellent and good rate of 87％.Conclusion Free flap transplantation can achieve satisfactory efficacy in treating Gustilo type ⅢB and ⅢC of tibia and fibula combined with soft tissue defects,with high limb salvage rate and good function recovery.

Objective To investigate the clinical therapy of early internal fixation combined with perforator flap for forearm open fractures of Gustilo types Ⅲ B & Ⅲ C.Methods A retrospective study was conducted of the 45 patients with forearm open fracture of Gustilo type ⅢB or Ⅲ C who had been treated from July 2012 to October 2016 at Department of Traumatic Orthopaedics,The Ninth People's Hospital of Wuxi.They were 26 men and 19 women,aged from 20 to 61 years (average,41 years).Twenty cases were Gustilo type ⅢB and 25 Gustilo type ⅢC.By AO classification,8 cases were type A,21 ones type B,and 16 ones type C.The wound size ranged from 4 cm × 3 cm to 36 cm × 8 cm.Thirty-three patients were treated by primary internal fixation plus secondary transfer with a perforator flap,12 ones by secondary internal fixation plus transfer with a perforator flap.The period from injury to secondary flap transfer ranged from 5 to 20 days (average,12 days).In this series,36 anterolateral thigh perforator flaps,5 latissimus dorsal muscular flaps and 4 lateral arm flaps were transferred.Results All the 45 free flaps survived with no deep infection or osteomyelitis.Partial necrosis happened at the distal ends of 2 latissimus dorsal muscular flaps which were cured by skin graft.Postoperative circulatory crisis happened after transfer of an anterolateral thigh perforator flap which survived with 5 cm skin necrosis at the distal end after successful surgical exploration.Superficial wound infection happened in 12 patients with no deep or bone infection.All the patients were followed up for 12 to 36 months (average,18.5 months).All the flaps were soft in texture,with varying degrees of pigmentation.The sensory recovery was S2 in 8 flaps,S3 in 29 flaps,and S4 in 8 flaps.Obvious scar hyperplasis was observed at the donor site in 5 cases while no obvious scar hyperplasis was observed in the other 40 ones.All the fractures got united after 4 to 14 months (average,8.6 months).Nonunion happened in 2 patients who were treated with autologous iliac graft 8 months after operation.By Anderson criteria,the curative efficacy was assessed as excellent in 15 cases,as good in 21,as fair in 7 and as poor in 2,yielding an excellent to good rate of 80.0％.Conclusion Early internal fixation combined with perforator flap transfe is an effective strategy for treatment of forearm open fractures with soft tissue defects of Gustilo types Ⅲ B &Ⅲ C,due to its advantages of shortened treatment period,possibility for early rehabilitation,decreased complications and satisfactory functional recovery.