Objective To monitor the antifungal resistance of clinical isolates of Candida spp.in the East China region.Methods MALDI-TOF MS or molecular methods were used to re-identify the strains collected from January 2018 to December 2022.Antifungal susceptibility testing was performed using the broth microdilution method.The susceptibility test results were interpreted according to the breakpoints of 2022 Clinical and Laboratory Standards Institute(CLSI)documents M27 M44s-Ed3 and M57s-Ed4.Results A total of 3 026 strains of Candida were collected,65.33％of which were isolated from sterile body sites,mainly from blood(38.86％)and pleural effusion/ascites(10.21％).The predominant species of Candida were Candida albicans(44.51％),followed by Candida parapsilosis complex(19.46％),Candida tropicalis(13.98％),Candida glabrata(10.34％),and other Candida species(0.79％).Candida albicans showed overall high susceptibility rates to the 10 antifungal drugs tested(the lowest rate being 93.62％).Only 2.97％of the strains showed dose-dependent susceptibility(SDD)to fluconazole.Candida parapsilosis complex had a SDD rate of 2.61％and a resistance rate of 9.42％to fluconazole,and susceptibility rates above 90％to other drugs.Candida glabrata had a SDD rate of 92.01％and a resistance rate of 7.99％to fluconazole,resistance rates of 32.27％and 48.24％to posaconazole and voriconazole non-wild-type strains(NWT),respectively,and susceptibility rates above 90％to other drugs.Candida tropicalis had resistance rates of 29.55％and 26.24％to fluconazole and voriconazole,respectively,resistance rates of 76.60％and 21.99％to posaconazole and echinocandins non-wild-type strains(NWT),and a resistance rate of 2.36％to echinocandins.Conclusions The prevalence and species distribution of Candida spp.in the East China region are consistent with previous domestic and international reports.Candida glabrata exhibits certain degree of resistance to fluconazole,while Candida tropicalis demonstrates higher resistance to triazole drugs.Additionally,echinocandins resistance has emerged in Candida albicans,Candida glabrata,Candida tropicalis,and Candida parapsilosis.

Objective:To investigate the long-term therapeutic effects and safety of renal denervation (RDN) on hypertensive patients with different cardiovascular risks, as well as its impact on adverse events, cardiovascular death and all-cause mortality.Methods:This was a single-center, single-arm, real-world retrospective study. Patients with refractory hypertension who underwent RDN at Tianjin First Central Hospital from July 6, 2011 to December 23, 2015 were enrolled and divided into either a high or intermediate-low risk group based on baseline cardiovascular risk. The treatment responsiveness of hypertensive patients with different cardiovascular stratification to RDN was assessed by comparing the results of office blood pressure, home blood pressure, and 24-h ambulatory blood pressure monitoring at 1, 5, and 11 years after RDN. Long-term safety of RDN was assessed by creatinine, and estimated glomerular filtration rate (eGFR) at 1 and 11 years after RDN. In addition, the total defined daily dose (DDD) of antihypertensive medications and the incidence of long-term adverse events, cardiovascular deaths, and all-cause deaths after RDN were followed up 11 years after RDN in person or by telephone.Results:A total of 62 patients with refractory hypertension, aged (50.2±15.0) years, of whom 35 (56.5%) were male, were included. There were 35 cases in high-risk group and 27 cases in low and medium risk group. The decrease in clinic systolic blood pressure (high risk vs. low-medium risk: (-38.0±15.1) mmHg vs. (-25.0±16.6) mmHg(1 mmHg=0.133kPa), P=0.002), home self-measured systolic blood pressure ((-28.4±12.7) mmHg vs. (-19.7±13.1) mmHg, P=0.011) and clinic systolic blood pressure 11 years after RDN ((-43.0±18.4) mmHg vs. (-27.8±17.9) mmHg, P=0.003) in the high-risk group was significantly higher than that in the low-medium risk group. The differences in heart rate and the decrease in total DDD number of antihypertensive drugs between the two groups were not statistically significant (all P>0.05). Creatinine and eGFR levels in the two groups at 1 and 11 years after RDN were not statistically significant when compared with the baseline values (all P>0.05). The cumulative cardiovascular mortality rate was 1.6% (1/62) and 8.1% (5/62), and the cumulative all-cause mortality rate was 3.2% (2/62) and 11.3% (7/62) at 5 and 11 years after RDN, respectively. The differences in the incidence rate of adverse events, cardiovascular mortality, and all-cause mortality rate between the two groups were not statistically significant (all P>0.05). Conclusions:RDN has long-term antihypertensive effect and good safety. Hypertensive patients who belong to the high-risk stratification of cardiovascular risk may respond better to RDN treatment.

OBJECTIVE: To explore the prevalence and characteristics of chromosomal abnormalities in abortuses during early pregnancy with single nucleotide polymorphism microarray (SNP-array). METHODS: For 520 abortuses, copy number variations (CNVs) in chorionic villi were analyzed with SNP-array. RESULTS: In 510 (98.1%) of the samples, the analysis was successful. Among these, 57.6% (294/510) of the samples were found to harbor clinically significant chromosomal abnormalities. 38.8% of the samples (198/510) had a normal result. 2.4% (12/510) of the samples harbored benign CNVs, and 1.2% (6/510) harbored variants of uncertain significance (VOUS). Aneuploidies, polyploidies, pathogenic CNVs and uniparental disomies (UPD) had accounted for 75.2% (221/294), 13.9% (41/294), 8.2% (24/294), and 2.7% (8/294) of the samples, respectively. 45,XO was the most common finding, which was followed by trisomy 16 and trisomy 22. 69,XXY was the most common polyploidy. CONCLUSION Chromosomal abnormalities are the main cause for early miscarriage, among which aneuploidies are most common. The prevalence of aneuploidies is significantly increased among women over 35. SNP-array analysis has the advantage of high success rate, high resolution and great accuracy, but the clinical significance of microdeletions/microduplications found by SNP-array can be difficult for interpretation.
Chorionic Villi ; Chromosome Aberrations ; Chromosome Disorders ; DNA Copy Number Variations ; Female ; Genetic Testing ; Humans ; Karyotyping ; Polymorphism, Single Nucleotide ; Pregnancy

Objective To investigate the value of next-generation sequencing (NGS) technique for genetic analysis of spontaneous abortion. Methods From January to June 2017, 154 patients who visited the First Affiliated Hospital of Zhengzhou University for spontaneous abortion were enrolled. All abortion tissue samples were analyzed by both NGS combined with short tandem repeat (STR) and single nucleotide polymorphism array (SNP-array). Results of the two methods were compared by Chi-square or Fisher's exact test. Results (1) Chromosomal abnormalities were detected in 109 of the 154 cases (70.7%), including 52 (47.7%) of numerical chromosomal abnormalities, 49 (45.0%) of structural chromosomal abnormalities, six (5.5%) of mosaicism, and two (1.8%) of uniparental disomy (UPD). In those 52 cases of numerical chromosome abnormalities, there were 45 of chromosome aneuploidy and seven of polyploidy. The top three numerical chromosomal abnormalities were 45,X (27.0%, 14/52), trisomy 22 (9.6%, 5/52) and trisomy 16 (7.7%, 4/52). Forty-nine structural abnormality cases carried 67 copy number variations (CNV), including 13 pathogenic CNV (pCNV, 19.4%), 24 variants of unknown clinical significance (35.8%) and 30 benign CNV (44.8%). In those 13 pCNVs, two were responsible for microdeletion and microduplication syndromes. (2) SNP-array was successful in 152 cases, but failed in two (1.3%) due to genomic DNA <200 ng. However, NGS technology was successful in all 154 cases and identified chromosomal abnormalities in the two cases that SNP-array had failed. No statistically significant difference was shown in the detection rate of chromosomal abnormalities between SNP-array and NGS technology [70.4% (107/152) vs 67.5% (104/154), χ2=0.293, P=0.588]. (3) No significant difference in the detection of chromosome aneuploidy (six cases in each group, 3.9% vs 3.9%) and mosaicism (45 cases in each group, 29.2% vs 29.6%) was found between NGS technology and SNP-array. Three cases of polyploidy (69, XXX) and two of UPD were identified by SNP-array, but not by NGS. When combined with STR, NGS was able to detect all three cases of polyploidy (69, XXX). (4) Forty-seven structural abnormality cases detected by SNP-array carried 53 CNVs, and 49 detected by NGS carried 67 CNVs. (5) NGS detected ten, three and one more CNVs than SNP-array did when the genome lengths were 100-<500, 500-<1 000 and ≥1 000 kb, respectively. Conclusions NGS can be used to detect chromosomal aneuploidy and mosaicism that can be identified by SNP-array with fewer limitations on total amount of genome. Moreover, CNVs that fail to be identified by SNP-array can also be detected by NGS. When combined with STR, NGS can effectively detect chromosomal polyploidy. Therefore, NGS could be a potential genetic analysis method for spontaneous abortion and of importance for genetic counseling.

Objective To investigate the value of the expression of Adenosine Triphosphate (ATP) binding cassette transporter A1 (ABCA1),peroxisome proliferator activated receptor γ(PPARγ) and sterol regulato-ry element binding protein (SREBP),adiponectin (ADPN) and liver X recepto α(LXRα) in type 2 diabetic pa-tients.Methods 71 patients with type 2 diabetes received in the hospital from June 2015 to June 2017 were se-lected as the observation group,and 60 healthy persons who underwent the health assessment from June 2015 to June 2017 were selected as the control group.Peripheral venous blood was collected from patients with an empty stomach in the morning,serum was isolated and serum human ADPN content were measured by radio-immunoassay.The levels of serum ABCA1,PPAR γ,SREBP and LXR α were measured by Enzyme linked im-munosorbent assay.Results The serum levels of ABCA1 and ADPN in the observation group were lower than those in the control group,while serum PPARγ SREBP and LXRα levels were higher than those in the control group (P< 0.05);the diagnostic sensitivity and specificity of ABCA 1+ PPARγ+ SREBP+ ADPN + LXRα were higher than those of single detection of ABCA1,PPARγ,SREBP,ADPN and LXRα.Conclusion The levels of ABCA1 and ADPN decreased in patients with type 2 diabetes,while the levels of PPAR γ,SREBP and LXR α was increased.The five joint diagnosis of ABCA1+ PPAR γ+SREBP+ADPN+LXR α has high sensitivity and specificity.It was of important clinical value and worth further application.

Objective: To investigate the value of single nucleotide polymorphism array (SNP-array) for fetuses with abnormal ultrasound findings. Method: A total of 904 fetuses with abnormal ultrasound findings were enrolled in this study from May 2015 to November 2017, and 434 (48.0%) cases received conventional karyotyping analysis at the same time. According to different abnormal ultrasound category, 904 cases were divided into 5 groups: 280 cases (31.0%) in single system structural anomalies, 31 cases (3.4%) in multiple system structural anomalies, 331 cases (36.6%) in single ultrasound soft marker abnormalities without structural anomalies, 107 cases (11.8%) in multiple soft marker abnormalities and 155 cases (17.2%) in structural abnormalities combined with soft markers abnormalities. Abnormal detection rates by SNP-array among 5 groups of abnormal ultrasound category were calculated. Result: (1) Total SNP-array results: 171 (19.0%) cases out of 904 cases analyzed by SNP-array, presented chromosomal abnormalities. Pathogenic copy number variants were detected in 27 cases (3.0%) and variants of unknown significance were detected in 81 cases (7.8%) . In addition, 7 cases (26.0%) were found with new mutation by parental validation. (2) SNP-array of 5 groups: among the 5 groups of abnormal ultrasound category, chromosomal abnormalities were identified by SNP-array in 19.3% (54/280) with single system structural abnormalities, 25.8% (8/31) with multiple system structural abnormalities, 13.9% (46/331) with single nonstructural anomalies, 19.6% (21/107) with multiple nonstructural anomalies and 27.1% (42/155) with structural abnormalities combined with nonstructural anomalies. The differences were significant (P=0.010) . No chromosome abnormalities was identified in single soft marker abnormalities, such as choroid plexus cysts, echogenic foci in the heart, single umbilical artery and pyelectasis. (3) Chromosomal abnormalities: the abnormal detection rate of aneuploidy chromosomal abnormalities by SNP-array increased with the maternal age, decreased with the gestational weeks (all P<0.05) . However, the pathogenic copy number variants and variants of unknown significance rates did not change with maternal age and gestational weeks (all P>0.05) . (4) SNP-array and karyotyping: 434 cases were analyzed by conventional karyotyping and SNP-array respectively, 10.3% (43/419) of which presented chromosomal abnormalities by conventional karyotyping and 18.7% (81/434) of which presented chromosomal abnormalities by SNP-array. Conclusions SNP-array could be a useful genetic analysis method in prenatal diagnosis for fetuses with abnormal ultrasound findings. For different abnormal ultrasound category, SNP-array has different detection rate. Compared with conventional karyotyping analysis, SNP-array can improve the detection rates for chromosomal abnormalities and find the chromosome abnormalities which can′t be detected by conventional karyotyping analysis. In clinical prenatal genetic counseling, SNP-array should be selected rationally in combination with the various abnormal ultrasound category.

OBJECTIVE:To study the effect of dihydromyricetin(DMY)on high glucose(HG)-induced glomerular mesangial cell(MCs)proliferation and fibronectin(FN)accumulation,and explore its mechanism for diabetic nephropathy glomerulosclero-sis. METHODS:Cells were divided into normal group(5.5 mmol/L glucose),HG group(30 mmol/L glucose),DMY low-concen-tration,medium-concentration,high-concentration groups (30 mmol/L glucose+22.5,45,90 μmol/L DMY). After incubating 48 h,MTT was used to detect the proliferative activity [reflected by the optical density(OD)value] of cells;molecular docking meth-od was adopted to conduct simulation analysis for DMY binding state with Smad2. Cells were divided into normal group(5.5 mmol/L glucose),HG group (30 mmol/L glucose),DMY group (30 mmol/L glucose+45 μmol/L DMY) and DMY control group (5.5 mmol/L glucose+45 μmol/L DMY). After incubating 5 h,Western blot was used to detect the expression levels of phosphorylated Smad2 (p-Smad2) and extracellular matrix protein FN. RESULTS:Results of MTT detection showed,compared with normal group,OD values in HG group were significantly increased(P<0.05);compared with HG group,OD values in DMY each con-centration group were significantly reduced(P<0.05). The Gibbs free energy(ΔG)of DMY and Smad2 protein was -5.64 kJ/mol, Ki was 73.53 μmol/L,and there were hydrogen bond donor and receptor binding in No. 465,464,461,458 amino acid residues. Results of Western blot showed,compared with normal group,expression levels of p-Smad2 and extracellular matrix protein FN in HG group were significantly increased (P<0.05);compared with HG group,expression levels of p-Smad2 and extracellular ma-trix protein FN in DMY group were significantly decreased(P<0.05). CONCLUSIONS:DMY inhibits HG-induced MCs prolifera-tion and improves diabetic nephropathy glomerulosclerosis by combining with Smad2 and inhibiting Smad2 phosphorylation to re-duce the extracellular matrix protein FN expression.

Objective To investigate the value of 320-detector row CT angiography for evaluating diabetic dorsalis pedis arterio-sclerotic disease.Methods 18 patients with diabetes and 20 healthy volunteers underwent 320-detector row CT angiography.The obtained thin-slice images were reformatted using maximum intensity projection,volume rendering,curved planar reformation and multiplanar reconstruction to evaluate the features of dorsalis pedis artery.Results 320-detector row CT angiography could exactly show the course,morphological features of the dorsalis pedis artery and the anatomical detail of arteriosclerotic disease.Arterioscle-rostic was found in variant degree in 18 cases.Among all,6 and 7 sides were slight arteriostenosis in proximal and distal segment,re-spectively.Moreover,2 and 3 sides were medium arteriostenosis in proximal and distal segment,respectively.The most common type of plaque property were mixed plaque.Conclusion 320-detector row CT angiography is of significant value in the assessment of arte-riosclerotic disease of dorsalis pedis arteries in the patients with diabetes.

Objective To verify the infantile amnesia phenomenon and discuss the effect of rapamycin injection on infantile amnesia and hippocampal neurogenesis.Methods (1) In experiment one,17-day-old (P17) and 60-day-old (P60) male Sprague-Dawley rats (n=84) were trained in a passive-avoidance task (they were randomly sub-divided into the shocked group and non-shocked group);all rats were tested immediately,2 and 7 days after training for memory retention.(2) In experiment two,P17 rats (n=84) were randomly divided into three groups:vehicle group,20 mg/kg rapamycin group and 40 mg/kg rapamycin group (they were sub-divided into the shocked group and non-shocked group);P17 rats were immediately injected with either vehicle or rapamycin after passive-avoidance training,and memory retention was evaluated 2 and 7 days after training.(3) In experiment three,P1 7 rats (n=48) were randomly divided into four groups:normal group,vehicle group,20 mg/kg rapamycin group and 40 mg/kg rapamycin group;immunofluorescence staining of BrdU/DCX was detected to observe the hippocampal neurogenesis at post-natal day 19 (P19) and post-natal day 24 (P24) after vehicle or rapamycin injection at P17 rats.Results In experiment one,P17 and P60 shocked rats exhibited comparable levels of crossing latencies when being tested immediately (P＞0.05),when being tested after 2 days and 7 days,P17 rats forgot more rapidly than P60 rats (P＜0.05);there were no age-related differences in crossing latencies between the non-shocked groups (P＞0.05).In experiment two,three shocked groups exhibited comparable levels of crossing latencies when being tested 2 days after training (P＞0.05),however,administration of rapamycin could enhance the memory retention 7 days after training;significant differences were noted between the vehicle group and each of the rapamycin treated groups (P＜0.05);there were no dose-related differences in crossing latencies among the non-shocked groups (P＞0.05).In experiment three,less BrdU-positive and BrdU/DCX-positive cells were found in the DG of the hippocampus at 20 mg/kg and 40 mg/kg rapamycin groups as compared with those in the vehicle group at P19 and P24 rats (P＜0.05).Conclusion The infantile amnesia phenomenon is verified,and rapamycin may enhance the memory retention in infant rats by reducing the hippocampal neurogenesis.