ObjectiveTo investigate the effect of icariin （ICA） on steroid-induced ferroptosis in bone microvascular endothelial cells （BMECs）. MethodsRat BMECs were selected and treated with 500 mg·L^-1 hydrocortisone for 1.5 h to establish a ferroptosis model of BMECs. The experimental cells were divided into a blank group， hormone group （500 mg·L^-1 hydrocortisone）， ICA group （500 mg·L^-1 hydrocortisone + 34 mg·L^-1 ICA）， and ferroptosis agonist group （500 mg·L^-1 hydrocortisone + 34 mg·L^-1 ICA + 2.7 mg·L^-1 erastin）. Cell viability was detected by CCK-8. The levels of ferrous ion， glutathione （GSH）， malondialdehyde （MDA）， superoxide dismutase （SOD）， and reactive oxygen species （ROS） were detected by related kit species. The ferroptosis-related proteins， such as glutathione peroxidase 4（GPX4）， ferritin light chain （FTL）， and transferrin receptor protein1 （sTfR） were detected by Western blot， as well as autophagy-related proteins including microtubule-associated protein 1 light chain 3B （LC3B）， Beclin1， B-cell lymphoma-2 （Bcl-2）， and Caspase-3. Results500 mg·L^-1 hydrocortisone intervention for 1.5 h could effectively induce ferroptosis in BMECs， and ferroptosis levels could reach a peak as the intervention continued. In terms of cellular antioxidant capacity， compared with those in the blank group， the cell vitality， GSH in the hormone group decreased significantly， and the levels of ROS， SOD， MDA， and ferrous ions were significantly increased （P<0.01）. Compared with those in the hormone group， the cell viability， GSH were significantly increased， and the levels of ROS， SOD， MDA， and ferrous ions were decreased in the ICA group （P<0.01）. Compared with those in the ICA group， the cell vitality， GSH in the ferroptosis agonist group decreased significantly， and the levels of ROS， SOD， MDA， and ferrous ions increased significantly （P<0.01）. In terms of the relationship between ferroptosis and autophagy， compared with the blank group， the hormone group had significantly increased expression levels of LC3B， sTfR， Beclin1， and FTL and significantly decreased expression levels of GPX4 （P<0.01）. Compared with the hormone group， The ICA group had significantly decreased expression levels of LC3B， sTfR， and FTL and significantly increased expression levels of Beclin 1 and GPX4 （P<0.01）. Compared with those in the ICA group， the expression levels of LC3B， sTfR， and FTL increased in the rapamycin group， and those of Beclin 1 and GPX4 decreased （P<0.01）. In terms of cell ferroptosis and apoptosis，compared with the blank group， the hormone group had significantly increased expression levels of FTL， sTfR and Caspase-3 and significantly decreased expression levels of GPX4， and Bcl-2 （P<0.01）. Compared with the hormone group， the ICA group had significantly decreased expression levels of FTL， sTfR and Caspase-3 and significantly increased expression levels of GPX4， and Bcl-2 （P<0.01）. Compared with those in the ICA group， the expression levels of FTL， sTfR and Caspase-3 in the ferroptosis agonist group were increased， and the expression levels of GPX4， and Bcl-2 were decreased （P<0.01）. In terms of cell function，compared with that in the blank group， the ability of cell migration and tube formation was significantly decreased in the hormone group （P<0.01）. Compared with that in the hormone group， the cell migration and tube formation ability in the ICA group were significantly increased （P<0.01）. ConclusionFerroptosis is involved in steroid-induced damage in BMECs. ICA can inhibit steroid-induced ferroptosis in BMECs， and the mechanism may be associated with the inhibition of ferroptosis by regulating autophagy.

Objective:To explore the prenatal ultrasound phenotype and genetic basis of two fetuses with Wolf-Hirschhorn syndrome (WHS).Methods:A retrospective analysis was conducted on the ultrasound imaging data of two fetuses suspected for WHS at the Prenatal Diagnostic Center of Qingyuan People′s Hospital in July 2017 and August 2019, respectively. Amniotic fluid samples of the two fetuses were subjected to chromosomal karyotyping and chromosomal microarray analysis (CMA). This study was approved by Medical Ethics Committee of the Qingyuan People′s Hospital (Ethics No. IRB-2022-064).Results:Prenatal ultrasound examination of the two fetuses had consistently revealed WHS-associated traits including intrauterine growth restriction (IUGR), craniofacial abnormalities and cardiovascular anomalies. Karyotyping analysis suggested that both fetuses had harbored cryptic chromosomal translocations involving partial deletion of 4p. And parental verification revealed that it was de novo for fetus 1 and paternal for fetus 2. CMA has confirmed that fetus 1 had an approximately 8.7 Mb deletion at 4p16.3p16.1 and a 6.8 Mb duplication at 8p23.1p23.1, whilst fetus 2 had a 20.05 Mb deletion at 4p16.3p15.31 and a 7.66 Mb duplication at 9p24.3p24.1. The karyotype of fetus 1 was determined as 46, XN, der(4)t(4; 8)(p16.1; p23.1)dn.arr[hg19]4p16.3p16.1(68345_8721580)×1, 8p23.3p23.1(158048_6933745)×3, and that of fetus 2 was determined as 46, XN, der(4)t(4; 9)(p15.3; p24)dpat.arr[hg19]4p16.3p15.31(68345_20116061)×1, 9p24.3p24.1(208454_7868292)×3. Conclusion:The 4p deletion is probably the main cause for the WHS phenotype in both fetuses. WHS should be suspected when IUGR, renal anomalies, craniofacial and cardiovascular abnormalities are detected upon prenatal ultrasound screening.

Objective To investigate characteristics of cognitive domain damage and risk factors of prodromal Parkinson's disease(pPD)patients with mild cognitive impairment(MCI).Methods A total of 82 pPD patients and 41 normal controls(normal control group)were included.According to MCI,pPD patients were divided into pPD-MCI group and pPD control group(pPD-NC group).The difference of unified Parkinson's disease rating scale Ⅲ(UPDRS Ⅲ),30 items non-motor symptom questionnaire(NMSQ),Hamilton depression scale(HAMD-24),Hamilton anxiety scale(HAMA),MMSE,Montreal cognitive assessment scale(MoCA),Chinese university of Hong Kong rapid eye movement sleep behavior disorder scale(RBDQ-HK),German Burghart sniff stick test and z-scores of each cognitive domain were compared among the groups.Logistic regression analysis was used to analyze the influencing factors of MCI.Results Compared with those in pPD-NC group,UPDRS Ⅲ score in pPD-MCI group was significantly increased,and MoCA score in pPD-MCI group was significantly decreased(P<0.05-0.01).There was no significant difference in NMSQ,HAMA,HAMD-24,MMSE,German Burghart sniff stick test and RBDQ-HK score between pPD-MCI group and pPD-NC group(all P>0.05).Compared with those in pPD-NC group,z-scores of attention,memory,visuospatial ability and language function subdomains in pPD-MCI group were significantly decreased(P<0.05-0.01).Multivariate Logistic regression analysis showed that high UPDRS Ⅲ score was an independent risk factor for Ppd-MCI(OR=0.799,95%CI:0.680-0.938,P=0.006).Conclusions The motor symptoms of Ppd-MCI patients are more severe than those of Ppd-NC patients.Memory,visuospatial ability and language function are the main cognitive impairment domains.High UPDRS Ⅲ score is an independent risk factor for Ppd cognitive impairment.

We aimed to evaluate the effectiveness and safety of single-course initial regimens in patients with low-risk gestational trophoblastic neoplasia (GTN). In this trial (NCT01823315), 276 patients were analyzed. Patients were allocated to three initiated regimens: single-course methotrexate (MTX), single-course MTX + dactinomycin (ACTD), and multi-course MTX (control arm). The primary endpoint was the complete remission (CR) rate by initial drug(s). The primary CR rate was 64.4% with multi-course MTX in the control arm. For the single-course MTX arm, the CR rate was 35.8% by one course; it increased to 59.3% after subsequent multi-course MTX, with non-inferiority to the control (difference -5.1%,95% confidence interval (CI) -19.4% to 9.2%, P = 0.014). After further treatment with multi-course ACTD, the CR rate (93.3%) was similar to that of the control (95.2%, P = 0.577). For the single-course MTX + ACTD arm, the CR rate was 46.7% by one course, which increased to 89.1% after subsequent multi-course, with non-inferiority (difference 24.7%, 95% CI 12.8%-36.6%, P < 0.001) to the control. It was similar to the CR rate by MTX and further ACTD in the control arm (89.1% vs. 95.2%, P =0.135). Four patients experienced recurrence, with no death, during the 2-year follow-up. We demonstrated that chemotherapy initiation with single-course MTX may be an alternative regimen for patients with low-risk GTN.
Antineoplastic Combined Chemotherapy Protocols/adverse effects* ; Dactinomycin/adverse effects* ; Female ; Gestational Trophoblastic Disease/drug therapy* ; Humans ; Methotrexate/therapeutic use* ; Pregnancy ; Retrospective Studies

Neuron-antibody mediated movement disorders are a class of movement disorders caused by abnormal immune responses to nerve auto-antigens,which could be paraneoplastic,infectious or idiopathic.The clinical manifestations of nervous system symptoms varied,and movement disorders can be used as part of various phenotypes,including chorea,dystonia,Parkinson's syndrome,myoclonus,stiffness syndrome,autoimmune encephalitis and cerebellar ataxia and so on.The clinical diagnosis of this disease is mainly based on the positive specific anti-central or peripheral nerve autoantibodies in serum or cerebrospinal fluid.The specificity of antibodies helps to predict associated cancer and its response to immunotherapy.Early and accurate diagnosis is very important because of the good therapeutic effect of early immunization.

Objective To grasp the current situation of drinking water-borne endemic fluorosis in Shandong Province,and provide a scientific basis for formulating preventive strategies.Methods In 2016,according to the "National Surveillance Scheme of Drinking Water-Borne Endemic Fluorosis" and "Shandong Provincial Surveillance Scheme of Drinking Water-Borne Endemic Fluorosis",10 counties (cities,districts) were selected in Shandong Province,3-5 villages were selected in each county (city,district),to investigate the operation of water improvement and fluorine reduction project and the water fluorine,and 1 tap water sample in each village was collected to detect water fluorine.At the same time,dental fluorosis examination was performed on all children aged 8-12 years old and fluorosis X-ray examination was performed on adults aged ≥25 years old.Water fluorine was detected by ion selective electrode method;the water fluorine was determined according to "Standards for Drinking Water Quality" (GB 5749-2006);criteria for diagnosis,grading and epidemic intensity of dental fluorosis were based on "Diagnosis of Dental Fluorosis" (WS/T 208-2011);the diagnostic and grading criteria for skeletal fluorosis were based on the "Diagnostic Criteria for Endemic Skeletal Fluorosis" (WS 192-2008).Results All the 32 villages in 10 counties (cities,districts) changed water,water improvement and fluorine reduction project ran normally,24 villages had qualified water fluorine (≤ 1.20 mg/L),and 8 villages had water fluorine exceeding the standard.The detection rate of dental fluorosis in children was 25.75％ (419/1 627),and the difference between 10 counties (cities,districts) was statistically significant (x2 =288.55,P ＜ 0.01);the dental fluorosis index was 0.48,and the defect rate was 2.77％ (45/1 627).The detection rate of skeletal fluorosis in adults was 4.72％ (72/1 525),and the difference between 10 counties (cities,districts) was statistically significant (x2 =18.83,P ＜ 0.01).Conclusions The problem of water fluorine exceeding the standard in water improvement and fluorine reduction project in Shandong Province is still serious.The condition of fluorosis has not been fully controlled,and the prevention and treatment measures need to be further strengthened.

OBJECTIVE: To report on the clinical pictures of 7 patients from a pedigree affected with X-linked adrenal hypoplasia congenita (XL-AHC) and hypogonadotropic hypogonadism (HH) and the underlying mutations. METHODS: Seven patients were identified from a four-generation pedigree affected with XL-AHC and HH. Their clinical features, endocrinological changes, treatment and drug response were recorded. The patients were subjected to next-generation sequencing, and the result was verified by Sanger sequencing. PolyPhen-2 was used for predicting the influence of the mutation on protein production. RESULTS: Three deceased patients had manifested adrenal insufficiency (AI) within one year after birth. Two died at 6 and one died at 12. The four survivors presented with salient clinical and endocrinological features of AHC and HH, adrenal and testicular atrophy, and renin-angiotensin compensation. Two adult patients had testicular micro-stone detected by ultrasound.One of them also had remarkable seminiferous tubule degeneration by biopsy. The patients were followed up for 0.5 to 10 years. All required hyper-physiological dose of hydrocortisone to stabilize their clinical condition. In three patients, gonadotropic or androgen replacement induced cardinal masculine development but with unsatisfactory testis growth and sperm production.Genetic analysis revealed a novel missense c.827A>C (p.Q276P) mutation in a hotspot region within a highly conserved domain. PolyPhen-2 predicted the mutation to be highly hazardous. CONCLUSION The novel p.Q276P mutation of the DAX1 gene probably underlies the XL-AHC and HH in this pedigree with variable clinical presentations in the patients.
Adrenal Insufficiency ; DAX-1 Orphan Nuclear Receptor ; genetics ; Humans ; Hypoadrenocorticism, Familial ; genetics ; Male ; Mutation ; Mutation, Missense ; Pedigree ; Repressor Proteins

Objective To screen and identify the in vivo-induced antigen Tp0462 of Treponema pallidum (Tp),and to evaluate its value for clinical serological diagnosis of syphilis.Methods Genomewide DNA was extracted from the Tp Nichols strain,and polymerase chain reaction (PCR) was performed to amplify the Tp0462 gene.A recombinant plasmid pET30a (+)-Tp0462 was constructed and transfected into the Escherichia coli (E.coli) Rosetta (DE3) strain.Recombinant protein Tp0462 was abundantly expressed,purified and identified.A total of 18 New Zealand rabbits were randomly and equally divided into 3 groups:viable Tp-incubating group incubated with viable Tp in the testes,inactived Tp-incubating group incubated with ultraviolet irradiation-killed Tp in the testes,and control group receiving no treatment.After incubation,blood samples were collected at different time points,and the sera were isolated for identification of characteristics of in vivo-induced antigen Tp0462.Enzyme-linked immunosorbent assay for Tp0462 (Tp0462-ELISA),Treponema pallidum particle agglutination assay (TPPA),preliminary syphilis screening ELISA and rapid plasma reagin (RPR) card test were applied in 336 clinical serum samples from patients with syphilis,so as to preliminarily evaluate the value of Tp0462 for the diagnosis of syphilis.Results The optimum conditions for expression of the recombinant plasmid Tp0462-pET30a(+) in E.coli Rosetta (DE3) strains were the treatment with 0.5 mmol/L isopropyl thiogalactoside (IPTG) on a shaker at 180 rpm for 4 hours.In the viable Tp-incubating group,the serum level of specific anti-Tp0462 antibody sharply increased from week 2,and went steady after week 5.However,the specific anti-Tp0462 antibody maintained a low level in the inactived Tp-incubating group and the control group.The viable Tp-incubating group showed a significantly higher level of specific anti-Tp0462 antibody compared with the inactived Tp-incubating group and the control group (both P ＜ 0.05),while no significant difference was observed between the inactived Tp-incubating group and the control group (P =0.256).The level of anti-Tp92 antibody was significantly higher in the viable Tp-incubating group and the inactived Tp-incubating group than in the control group (P ＜ 0.05),while there was no significant difference between the viable Tp-incubating group and the inactived Tp-incubating group (P =0.127).Compared with TPPA,the sensitivity,specificity,consistency rate and area under the curve (AUC) of Tp0462-ELISA for the diagnosis of syphilis were 91.7％,98.8％,95.2％ and 0.997 respectively.Tp0462-ELISA was consistent to preliminary syphilis screening ELISA and RPR with a Kappa coefficient of 0.846 and 0.293,respectively.Conclusion Tp0462-ELISA has shown evidently higher sensitivity and specificity in the serodiagnosis of syphilis,and Tp0462 can serve as promising antigens for the diagnosis of syphilis.

Objective To study comparatively the performance of rapid plasma reagin (RPR) test in syphilis detection among pregnant women and non-pregnant women to provide reference for detecting syphilis in pregnant women.Methods The women aged 20-40 years old were selected and divided into the pregnant group and the non-pregnant group.RPR and treponema pallidum particle assay(TPPA) were simultaneously adopted to conduct the syphilis detection.The positive cases were judged by the TPPA detection results combined with the contact history,clinical symptoms and treatment situation.The results were compared with those by RPR for determining the false negative and false positive in RPR.The false negative rate and false positive rate of RPR detection results were analyzed in the two groups.Results Among 117 pregnant women,15 cases were false negative in RPR and 9 cases were false positive in RPR;among 755 non-pregnant women,there were 44 cases of false negative RPR results and 8 cases of false positive RPR results.The false negative rates in the pregnant group and non-pregnant group were 25.0% and 8.8% respectively,the difference was statistically significant(χ2=14.739,P<0.05);the false positive rates in the pregnant group and non-pregnant group were 15.7% and 3.1% respectively,the difference was statistically significant (χ2=14.722,P<0.05).Conclusion There are many factors affecting RPR for detection syphilis,pregnant women are the specific group,so higher false positive rate and false negative rate exist than non-pregnant women,the detection results should be comprehensively judged by combining with clinical symptoms and disease history,if necessary,combining with other syphilis detection method for avoiding missed diagnosis and misdiagnosis.

Objective To investigate the clinical distribution and antimicrobial resistant characteristics of Pseudomonas aeruginosa(PA)in the northern area of Guangdong.Provide the reference for clinical to prevent infection and reasonable choice of antibiotics and reduce the production of drug resistance strains.Methods The separation and identification of PA were performed by conventional methods during the data of drug 2013 and 2014.The data of sensitivity test of PA were analyzed by WHONET 5.6 and SPSS19.0 softwares.Results The 584 strains PA were mainly distributed in ICU,department of orthopaedics and respiratory medicine.Specimens were mainly from sputum and wound secretion.The detection of PA to 12 antibacterial agents showed different resistance.The antimicrobial with highest resistance was the gentamicin and lower resistance rates to fluoroquinolones,carbapenems,enzyme inhibitors.And a downward trend was shown in drug resistance to CIP,FEP,LEV,SCF.Conclusion PA mainly cause lung and wound infection,especially those old patients that come from ICU,department of orthopaedics and respiratory medicine.Although the drug resistance rates of PA to the commonly used antibiotics are relatively low,The clinicians should reasonably use antibiotics so as to reduce the resistant strains,especially the produce of MDR-PA and PDR-PA.