Objective To establish a rapid detection method for zika virus based on direct amplification re-al-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)technique.Methods A direct amplification RT-PCR technique for the rapid detection of zika virus in 5 samples(whole blood,serum,saliva,throat swab and urine)was established by using a special function DNA polymerase and a preferred PCR enhancer.Results The detection limits of the 5 samples were 103 PFU/mL in serum,102 PFU/mL in urine,throat swab,and saliva,and 104 PFU/mL in whole blood.The coefficient of goodness-fit of stand-ard curves was above 0.98,and the amplification efficiency was 90％-110％.Zika virus nucleic acid was suc-cessfully amplified,but non-zika virus nucleic acid was not amplified.Based on the repeatable detection of sam-ples from urine,whole blood,and saliva,the variation coefficient of 6 repeated Ct values at 106 PFU/mL and 102 PFU/mL concentrations were all＜5％.The zika virus detection method established by the direct amplifi-cation RT-PCR technique was consistent with the detection results of conventional RT-PCR technique.Only two serum samples were detected in eight zika virus samples,and the remaining 62 non-zika virus samples and 12 negative samples were not amplified.Conclusion A rapid detection method for zika virus based on direct ampli-fication RT-PCR technique is successfully established.The method is simple,rapid,sensitive and specific.

BACKGROUND:Resveratrol is a natural antioxidant extracted from plants.Its mechanism of improving exercise-induced fatigue mainly focuses on the protective effect against oxidative stress and inflammation.In this study,the protective mechanism of resveratrol on exercise-induced fatigue was mainly discussed from the perspective of gluconeogenesis. OBJECTIVE:To investigate the effect of resveratrol on gluconeogenesis in exercise-induced fatigue rats. METHODS:After 1 week of adaptive training,male Sprague-Dawley rats were randomly divided into 4 groups with 12 rats in each group:blank control group,resveratrol group,exercise group,resveratrol + exercise group.Weight-bearing swimming training was used to simulate long-term medium-high intensity exercise.After swimming with a weight of 5%for 1 hour every day,50 mg/kg resveratrol solution or the same volume of dimethyl sulfoxide solvent were given orally,6 days a week,for a total of 6 weeks.Samples were collected 24 hours after the last exercise,and the levels of urea nitrogen,creatine kinase,blood glucose,liver glycogen and lactic acid and pyruvate in liver tissue were detected by the kit.The activity of phosphoenolpyruvate carboxykinase was detected by microassay,and the activity of glucose-6-phosphatase was detected by enzyme-linked immunosorbent assay.Real-time fluorescence quantitative PCR was used to detect the gene expression of silent information regulator 1,cAMP-response element binding protein and peroxisome proliferator-activated receptor-γ coactivator-1α. RESULTS AND CONCLUSION:In the exercise group,plasma urea nitrogen and creatine kinase levels of rats were significantly increased(both P<0.05),liver lactate and pyruvate levels and lactate/pyruvate ratio were significantly increased(all P<0.01),and blood glucose and liver glycogen contents were significantly decreased(both P<0.01).Resveratrol supplementation could effectively improve the above conditions.Exercise significantly decreased the activities of phosphoenolpyruvate carboxykinase and glucose-6-phosphatase(P<0.01,P<0.05),and resveratrol supplementation significantly increased the activity of phosphoenolpyruvate carboxykinase in liver tissue(P<0.01).The mRNA expression levels of silent information regulator 1,cAMP-response element binding protein and peroxisome proliferator-activated receptor-γ coactivator-1α in liver tissue of the exercise group were significantly decreased(all P<0.01),while resveratrol supplementation could significantly increase the gene expression levels of this pathway.To conclude,resveratrol can relieve exercise-induced fatigue caused by long-term medium-high intensity exercise,and its mechanism may be related to up-regulating the gluconeogenesis regulatory pathway,improving rate-limiting enzyme activity,promoting liver gluconeogenesis,and increasing blood glucose and liver glycogen levels.

BACKGROUND:Studies have shown that resveratrol can relieve exercise-induced fatigue and protect the heart,but its action mechanism needs further study. OBJECTIVE:To explore the protective effect and regulatory mechanism of resveratrol on ventricular remodeling in exercise-induced fatigue rats. METHODS:Totally 48 male Sprague-Dawley rats were randomly divided into four groups,with 12 rats in each group.Rats in the blank control group were fed conventionally.After one week of adaptive training,rats in the exercise-related fatigue group and exercise-related fatigue with resveratrol supplement group were trained by 6-week weight-bearing swimming(5%body mass lead block fixed in the tail,70%-80%maximal oxygen uptake intensity),6 days a week,60 minutes a day.Rats in the resveratrol supplement group and exercise-related fatigue with resveratrol supplement group were given resveratrol(50 mg/kg per day)by gavage one hour after exercise intervention.Blank control group and exercise-related fatigue group were given the same volume of 2%dimethyl sulfoxide,6 days a week,once a day for 6 weeks.The body mass and heart mass of the rats were measured 24 hours after the last intervention.Plasma creatine kinase isoenzyme,cardiac troponin 1,pyruvate dehydrogenase and uncoupling protein 1 levels in myocardial tissue were determined by ELISA.The mRNA expression levels of ventricular remodeling-related factor Foxp1,transforming growth factor β1 and endothelin 1 were detected by RT-PCR. RESULTS AND CONCLUSION:Compared with the blank control group,the body mass of rats decreased and the heart mass increased in the exercise-related fatigue group(P＜0.05).Compared with the exercise-related fatigue group,the body mass and heart mass of the rats reduced in the exercise-related fatigue with resveratrol supplement group(P＜0.05).Compared with the blank control group,the levels of creatine kinase isoenzyme,cardiac troponin 1 and uncoupling protein 1 increased(P＜0.01),and the level of pyruvate dehydrogenase decreased(P＜0.01)in the exercise-related fatigue group.Compared with the exercise-related fatigue group,the levels of creatine kinase isoenzyme,myocardial troponin 1 and uncoupling protein 1 decreased(P＜0.05),and the level of pyruvate dehydrogenase increased(P＜0.05)in the exercise-related fatigue with resveratrol supplement group.Compared with the blank control group,the expression of the Foxp1 gene decreased(P＜0.01),and the expression of transforming growth factor β1 and endothelin 1 gene increased(P＜0.01)in the myocardium of the exercise-related fatigue group.Compared with the exercise-related fatigue group,the expression of the Foxp1 gene in the myocardium of the exercise-related fatigue with resveratrol supplement group increased(P＜0.01),while the expression of the transforming growth factor β1 and endothelin 1 gene decreased(P＜0.05).It is suggested that exercise-induced fatigue can promote myocardial adaptability and cause compensatory hypertrophy.Resveratrol can improve myocardial injury and energy metabolism and delay ventricular energy remodeling in rats.This effect may be related to the regulation of Foxp1/transforming growth factor β1/endothelin 1 signaling pathway.

Ovarian tissue cryopreservation and transplantation is the only way to preserve fertility for female cancer patients in prepubertal ages and those who cannot delay radiotherapy or chemotherapy.However,the success rate of cryopreservation and transplantation of ovarian tissue is still low at present due to the risk of ischemia and hypoxia of the grafted tissues.Abnormal activation of primordial follicles and ischemia-reperfusion injury after blood supply recovery also cause massive loss of follicles in grafted ovarian tissues.Various studies have explored the use of different drugs to reduce the damage of follicles during freezing and transplantation as well as to extend the duration of endocrine and reproductive function in patients with ovarian transplantation.For example,melatonin,N-acetylcysteine,erythropoietin or other antioxidants have been used to reduce oxidative stress;mesenchymal stem cells derived from different tissues,basic fibroblast growth factor,vascular endothelial growth factor,angiopoietin 2 and gonadotropin have been used to promote revascularization;anti-Müllerian hormone and rapamycin have been used to reduce abnormal activation of primordial follicles.This article reviews the research progress on the main mechanisms of follicle loss after ovarian tissue transplantation,including hypoxia,ischemia-reperfusion injury and associated cell death,and abnormal activation of follicles.The methods for reducing follicle loss in grafted ovarian tissues are further explored to provide a reference for improving the efficiency of ovarian tissue cryopreservation and transplantation.

A 15-year-old female with Hodgkin's lymphoma underwent ovarian tissue cryopreservation for preserving fertility in Reproductive Department of Sir Run Run Shaw Hospital,Zhejiang University School of Medical after receiving one course of chemotherapy.During the ovarian tissue cryopreservation,one MⅡmature oocyte and three germinal vesicle oocytes were found.The three immature oocytes underwent in vitro maturation but failed.Ultimately,one mature oocyte and 12 ovarian cortex slices were cryopreserved using vitrification.This case indicates that for patients with established gonadal axis feedback,ovarian tissue cryopreservation may not be the only method for fertility preservation.It is advisable to consider ovarian stimulation and oocyte retrieval for oocyte cryopreservation.Alternatively,for individuals in the ovulation phase of their menstrual cycle,attempting oocyte retrieval before ovarian tissue cryopreservation to obtain mature oocytes from the natural cycle,followed by oocyte cryopreservation,may enhance the likelihood of successful fertility preservation.

Purpose To study the clinical and pathological features of angiomatoid fibrous histocytoma(AFH)and to ex-plore its diagnosis,differential diagnosis and prognosis.Meth-ods The clinicopathological and follow-up data were analyzed in 14 cases of AFH,and the literatures were reviewed.Results There were 11 males and 3 females.The age ranged from 11 months to 12 years and 11 months,with average 5.9 years.3 cases were located in limbs,and 5 cases in trunk,5 cases in head and neck region,and 1 of intracranial tumor.Histological-ly,14 cases were composed of fibrous capsules and lymphocyte sheaths,and cell nucleus were vacuolar,forming fascicles with focal whirling and synteny.Intralesional pseudoangiomatous spaces were noted in 9 cases.Calcification was found in 2 ca-ses.2 cases showed high mitotic acticity(11/10 HPF).Scle-rosing and/or myxoid stroma was seen in 3 cases.Tumors were immunopositive for desmin(10/14),EMA(12/14),CD99(12/14),SMA(9/12),ALK(7/8),and the average of Ki67 index was 16％.7 cases harbored EWSR1 rearrangenent(part-ner gene not identified),2 cases had EWSR1-ATF1 fusion and 2 EWSR1-CREB1 fusion.Clinical follow-up information was a-vailable for 14 cases(average 46 months).All the 14 cases were alive without recurrence and metastasis.Conclusion AFH is a borderline or low-grade malignant tumor,often demon-strates indolent behavior in children,but rarely recurs and me-tastasizes.The diagnosis and differential diagnosis require a comprehensive analysis of clinical features,histopathologic changes,immunohistochemical finding and EWSR1 or FUS gene detection results.

Objective:To investigate the clinicopathological features, immunophenotype and molecular genetic characteristics of congenital spindle cell/sclerosing rhabdomyosarcoma.Methods:Sixteen cases (including 10 consultation cases) of congenital spindle cell/sclerosing rhabdomyosarcoma diagnosed at the Beijing Children′s Hospital, Capital Medical University, Beijing China, from April 2017 to January 2022 were collected. These cases were evaluated for clinical profiles, histomorphological features, immunophenotype and molecular characteristics.Results:Among the 16 patients, 9 were male and 7 were female. Five cases were present during maternal pregnancy and 11 cases were found immediately after birth. The tumors were located in the chest wall, low back, retroperitoneum, extremities or perineum. The tumors consisted of fasciculated spindle-shaped cells with localized mesenchymal sclerosis and vitreous metaplasia. Immunohistochemistry showed that the tumor cells expressed Desmin, Myogenin, MyoD1, SMA, CD56 and ALK to varying degrees, but not other markers such as CD34, CD99, pan-TRK, S-100 and BCOR. FISH analyses with NCOA2 (8q13) and VGLL2 (6q22) gene breakage probes revealed a breakage translocation in chromosome NCOA2 (8q13) in 4 cases (4/11). In the 6 cases subject to sequencing, a mutation at the p.L122R locus of MYOD1 gene was detected in 1 case (1/6). Two cases were examined by electron microscopy, which showed bundle-arranged myofilaments with some primitive myofilament formation. Five cases were resected with simple surgery, 2 cases were biopsied and followed up with observation only, and 9 cases were treated with surgery and adjuvant chemotherapy. Follow-up was available in 12 cases. At the end of the follow-up, 2 of the 12 patients developed local recurrences and 2 patients survived with disease.Conclusions:Congenital spindle cell/sclerosing rhabdomyosarcoma is a rare subtype of congenital rhabdomyosarcoma. It more commonly occurs in the chest, back and lower limbs of infants than other sites. NCOA2/VGLL2 gene fusion seems to be the most common genetic change. Its prognosis is better than other subtypes of rhabdomyosarcoma and those in adolescents and adults with the same subtype. Analysis and summary of its clinicopathological features can help differentiate it from other soft tissue tumors in infants and children and provide the information for appropriate treatments.

Prone position ventilation(PPV)is an essential therapeutic intervention for patients with refractory hypoxemic respiratory failure.The outbreak of coronavirus disease 2019(COVID-19)has further elevated the significance of PPV in clinical practice.Early enteral nutrition(EN)is considered a crucial measure for improving outcomes in critically ill patients.However,there remains considerable controversy regarding the implementation of EN during PPV.This review summarizes the research progress on the tolerance and safety of EN during PPV before and after the COVID-19 outbreak.It also highlights key strategies to enhance EN tolerance in prone-positioned patients,providing a clinical reference for improving patient management.

Objective To analyze the changes of serum humoral immunity related indicators, including immunoglobulins (Ig), complement and C-reactive protein (CRP) in patients with occupational medicamentose-like dermatitis due to trichloroethylene (OMDT) at different stages. Methods A total of 131 OMDT patients were selected as the study subjects using a retrospective analysis method, and liver function and humoral immunity related indicators were collected for analysis in the early stage and the recovery stage of the disease. Results The abnormality of liver function among study subjects was 89.3% (117/131) in the early stage of the disease, with mild, moderate, and severe liver damage accounting for 61.8%, 5.3%, and 22.1%, respectively. The levels of serum alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transferase, total bilirubin, alkaline phosphatase, total bile acid, IgG and CRP in the early stage increased ( all P<0.01), while the level of serum cholinesterase and C3 decreased (both P<0.01) compared with the recovery stage. However, there was no significant difference in the level of serum albumin, globulin, IgA, IgM and C4 or albumin/globulin ratio between the early stage and the recovery stage (all P>0.05). The level of patients′ serum IgA in the group with severe liver function injury was lower than that in the group with mild to moderate liver function injury in the recovery stage (P<0.05). The level of serum C3 in the patients with severe liver function injury was lower than that in the group with mild to moderate liver function injury in the early stage (P<0.05). Conclusion Serum IgG and CRP levels increased and C3 level decreased in OMDT patients at the early stage of the disease, which was correlated with the degree of liver function injury. It is suggested that humoral immune response is involved in the autoimmune liver injury in OMDT patients.

Objective To analyze the expression of microRNA-451 (miR-451) in children with microcytic anemia and chronic infectious anemia, and to explore the correlation between miR-451 and erythrocyte-related parameters and its diagnostic value for anemia in children. Methods The expression levels of miR-451 in serum were detected in children with microcytic anemia (n=80), chronic infectious anemia (n=80), and healthy controls (n=80) using real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The correlation between serum miR-451 and erythrocyte-related parameters[red blood cell distribution width (RDW), red blood cells (RBC), mean corpuscular hemoglobin concentration (MCHC), hemoglobin (Hb), mean corpuscular hemoglobin content (MCH), mean corpuscular volume (MCV)]was analyzed using Pearson correlation analysis. The receiver operating characteristic (ROC) curve was plotted to analyze the diagnostic value of miR-451 for microcytic anemia, chronic infectious anemia, and the severity of anemia. Multivariate Logistic regression analysis was used to investigate the influencing factors for the occurrence of microcytic anemia and chronic infectious anemia in children. Results The expression levels of serum miR-451 in the microcytic anemia group and chronic infectious anemia group were lower than those in the healthy control group(P < 0.05). The miR-451 was negatively correlated with RDW (r=-0.388, P < 0.05), and positively correlated with Hb, RBC, MCHC, MCH, and MCV (r=0.402, 0.393, 0.391, 0.360, 0.323, P < 0.05). ROC curve analysis showed that the areas under the curve of serum miR-451 for diagnosing microcytic anemia and chronic infectious anemia was 0.718 and 0.735, respectively, which was higher than that of erythrocyte-related parameters such as RDW, Hb, RBC, MCHC, MCH, and MCV (P < 0.05). Multivariate Logistic regression analysis showed that body mass index, RDW, Hb, RBC, MCHC, MCH, MCV, and miR-451 were influencing factors for the occurrence of microcytic anemia and chronic infectious anemia in children (P < 0.05). With the aggravation of anemia in children, the expression level of serum miR-451 gradually decreased (P < 0.05). ROC curve analysis showed that the level of serum miR-451 had a high diagnostic value for the severity of anemia in children. Conclusion The expression of miR-451 is reduced in serum of children with microcytic anemia and chronic infectious anemia, and its expression level is correlated with erythrocyte-related parameter levels. Serum miR-451 has a high diagnostic value for microcytic anemia, chronic infectious anemia, and the severity of anemia.