This study was conducted retrospectively on a cohort of 68 patients with steroid 5 α-reductase 2 (SRD5A2) deficiency and 46,XY disorders of sex development (DSD). Whole-exon sequencing revealed 28 variants of SRD5A2 , and further analysis identified seven novel mutants. The preponderance of variants was observed in exon 1 and exon 4, specifically within the nicotinamide adenine dinucleotide phosphate (NADPH)-binding region. Among the entire cohort, 53 patients underwent initial surgery at Sichuan Provincial People's Hospital (Chengdu, China). The external genitalia scores (EGS) of these participants varied from 2.0 to 11.0, with a mean of 6.8 (standard deviation [s.d.]: 2.5). Thirty patients consented to hormone testing. Their average testosterone-to-dihydrotestosterone (T/DHT) ratio was 49.3 (s.d.: 23.4). Genetic testing identified four patients with EGS scores between 6 and 9 as having this syndrome; and their T/DHT ratios were below the diagnostic threshold. Furthermore, assessments conducted using the crystal structure of human SRD5A2 have provided insights into the potential pathogenic mechanisms of these novel variants. These mechanisms include interference with NADPH binding (c.356G>C, c.365A>G, c.492C>G, and c.662T>G) and destabilization of the protein structure (c.727C>T). The c.446-1G>T and c.380delG variants were verified to result in large alterations in the transcripts. Seven novel variations were identified, and the variant database for the SRD5A2 gene was expanded. These findings contribute to the progress of diagnostic and therapeutic approaches for individuals with SRD5A2 deficiency.
Humans ; 3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics* ; Disorder of Sex Development, 46,XY/blood* ; Male ; Membrane Proteins/genetics* ; Child, Preschool ; Child ; Retrospective Studies ; Adolescent ; Female ; Mutation ; Testosterone/blood* ; Infant ; Dihydrotestosterone/blood*

OBJECTIVE: To reveal the neuroprotective effect and the underlying mechanisms of a mixture of the main components of Panax notoginseng saponins (TSPN) on cerebral ischemia-reperfusion injury and oxygen-glucose deprivation/reoxygenation (OGD/R) of cultured cortical neurons. METHODS: The neuroprotective effect of TSPN was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, flow cytometry and live/dead cell assays. The morphology of dendrites was detected by immunofluorescence. Middle cerebral artery occlusion (MCAO) was developed in rats as a model of cerebral ischemia-reperfusion. The neuroprotective effect of TSPN was evaluated by neurological scoring, tail suspension test, 2,3,5-triphenyltetrazolium chloride (TTC) and Nissl stainings. Western blot analysis, immunohistochemistry and immunofluorescence were used to measure the changes in the Akt/mammalian target of rapamycin (mTOR) signaling pathway. RESULTS: MTT showed that TSPN (50, 25 and 12.5 µ g/mL) protected cortical neurons after OGD/R treatment (P<0.01 or P<0.05). Flow cytometry and live/dead cell assays indicated that 25 µ g/mL TSPN decreased neuronal apoptosis (P<0.05), and immunofluorescence showed that 25 µ g/mL TSPN restored the dendritic morphology of damaged neurons (P<0.05). Moreover, 12.5 µ g/mL TSPN downregulated the expression of Beclin-1, Cleaved-caspase 3 and LC3B-II/LC3B-I, and upregulated the levels of phosphorylated (p)-Akt and p-mTOR (P<0.01 or P<0.05). In the MCAO model, 50 µ g/mL TSPN improved defective neurological behavior and reduced infarct volume (P<0.05). Moreover, the expression of Beclin-1 and LC3B in cerebral ischemic penumbra was downregulated after 50 µ g/mL TSPN treatment, whereas the p-mTOR level was upregulated (P<0.05 or P<0.01). CONCLUSION TSPN promoted neuronal survival and protected dendrite integrity after OGD/R and had a potential therapeutic effect by alleviating neurological deficits and reversing neuronal loss. TSPN promoted p-mTOR and inhibited Beclin-1 to alleviate ischemic damage, which may be the mechanism that underlies the neuroprotective activity of TSPN.
Animals ; Beclin-1 ; Brain Ischemia/metabolism* ; Glucose ; Infarction, Middle Cerebral Artery/drug therapy* ; Mammals/metabolism* ; Neuroprotection ; Neuroprotective Agents/therapeutic use* ; Oxygen ; Panax notoginseng ; Proto-Oncogene Proteins c-akt/metabolism* ; Rats ; Reperfusion Injury/metabolism* ; Saponins/therapeutic use* ; TOR Serine-Threonine Kinases/metabolism*

AIM: To observe the changes of the morphology and structure of macula, blood flow density of macula and optic disc, as well as retinal nerve fiber layer thickness by optical coherence tomography(OCT)and optical coherence tomography angiography(OCTA)in patients with rhegmatogenous retinal detachment(RRD)under went sclera buckling(SB). METHODS: As a cross-sectional case-controlled study, 25 patients(25 eyes)were diagnosed with RRD in the department of ophthalmology, Ganzhou People's Hospital from July 2014 to March 2021. The differences in the vascular density(VD)of superficial vessel cluster(SVC), the VD of deep vessel cluster(DVC)of macula, the SVC-VD of optic disc, retinal nerve fiber layer(RNFL), central macular thickness(CMT), subfoveal choroidal thickness(SFCT)and outer structure of macula between the affected eyes and healthy eyes at the last post-operative follow-up were compared, and the correlation indicators affecting best corrected visual acuity(BCVA, LogMAR)of the affected eyes at the last follow-up were analyzed. RESULTS: There were no statistically significant in SVC-VD and DVC-VD of macula, SVC-VD of optic disc, RNFL, CMT, SFCT between the affected eyes and healthy eyes at the last post-operative follow-up(all P>0.05); At the last follow-up visit, the comparison of macular outer structure on OCT between the affected eyes and the healthy eyes showed that the light band integrity of the external limiting membrane(ELM), myoid zone(MZ), ellipsoid zone(EZ)and outer segment of photoreceptor(OS)had no statistically significant difference(all P>0.05), while the light band integrity of interdigitation zone(IZ)had significant difference(P=0.014); The difference of BCVA(LogMAR)between the affected eyes and the healthy eyes at the last follow-up was statistically significant(P=0.002). There was significant correlation between BCVA(LogMAR)of affected eyes at the last post-operative follow-up and the presence or absence of macular involvement, the correlated with SVC-VD of optic disc, the integrity of the light bands of ELM, MZ, EZ, OS and IZ on the outer structure of macula, it was positively correlated with the presence or absence of macular involvement(rs=0.401, P=0.047)and it was negatively correlated with SVC-VD of optic disc, the integrity of the light bands of ELM, MZ, EZ, OS and IZ on the outer structure of macula(all P<0.05).CONCLUSION: The OCT and OCTA can be used to observe fundus changes after SB surgery for RRD to obtain long-term follow-up information related to vision prognosis, and visual prognosis depends on the recovery of retinal outer structure, and the integrity of IZ structure is more important for visual recovery; The SVC-VD of optic disc is correlated with visual prognosis, and whether it was correlated with intraocular pressure require further observation and verification with postoperative continuous data.

Background::Understanding the characteristics of newly diagnosed primary human deficiency virus-1 (HIV-1) infection in the context of the post-antiretroviral therapy era and HIV drug prophylaxis is essential for achieving the new target of 95-95-95-95 by 2025. This study reported the characteristics of newly diagnosed primary HIV-1 infection in Shenzhen.Methods::This is a real-world retrospective study. Eighty-seven newly diagnosed primary HIV-1-infected patients were recruited from January 2021 to March 2022 at the Third People’s Hospital of Shenzhen. Demographic, epidemiological, diagnostic, drug resistance, and medical data were described and analyzed.Results::Overall, 96.6% (84/87) of the newly identified primary HIV-1-infected patients were male, including 88.5% (77/87) men have sex with men (MSM), with a median age of 29.0 years (Q 1-Q 3: 24.0-34.0 years); of these, 85.1% (74/87) reported high-risk sexual behaviors with casual partners. The rate of condom usage was only 28.7% (25/87). The overall rate of pre-exposure prophylaxis (PrEP) and post-exposure prophylaxis (PEP) was 8.0% (7/87, including 4 PrEP and 3 PEP cases) around the potential exposure, although 41.4% of the patients had prior awareness of such interventions. Moreover, only 19.5% (17/87) had previously used PrEP or PEP. Of those, 58.8% (10/17) of the patients obtained drugs from the internet, and only 35.3% (6/17) reported good compliance. A total of 54.0% (47/87) of subjects were diagnosed by the HIV nucleic acid test. Acute retroviral syndrome appeared in 54.0% (47/87) of patients. The prevalence of transmitted drug resistance (TDR) mutation was 33.9% (19/56), including 6 (10.7%) against nucleoside reverse transcriptase inhibitor (NRTI) plus non-nucleoside reverse transcriptase inhibitor (NNRTI), 8 (14.3%) against NNRTI, and 5 (8.9%) against protease inhibitor (PI) only. Conclusions::Owing to the low utilization rate and incorrect usage of PrEP and PEP, massive efforts are needed to promote HIV-preventive strategies in the MSM population. The extremely high prevalence of TDR mutation in this population implies the need for future pretreatment drug resistance surveillance.

OBJECTIVE: To study the association of copy number of SMN1 and SMN2 with clinical phenotypes in children with spinal muscular atrophy (SMA). METHODS: A total of 45 children with SMA were enrolled. Multiplex ligation-dependent probe amplification was used to measure the gene copy numbers of SMN1 and SMN2. The association of copy number of SMN1 and SMN2 with clinical phenotypes was analyzed. RESULTS: Of the 45 children with SMA, 42 (93%) had a homozygous deletion of SMN1 exons 7 and 8, and 3 (7%) had a deletion of SMN1 exon 7 alone. No association was found between SMA clinical types and the deletion types of SMN1 exons 7 and 8 (P>0.05). There was a significant difference in the distribution of SMN2 gene copy numbers between the children with SMA and the healthy children (P<0.05). The children with SMA usually had two or three copies of SMN2 gene, while the healthy children usually had one or two copies of SMN2 gene. There was a significant difference in the distribution of SMN2 copy numbers among the children with different SMA clinical types (P<0.05). The children with two copies of SMN2 gene had a significantly lower age of onset than those with three or four copies. Most of the children with type I SMA had two or three copies of SMN2 gene. Most of the children with type II SMA had three copies of SMN2 gene. Most of the children with type III SMA had three or four copies of SMN2 gene. Children with a higher copy number of SMN2 gene tended to have an older age of onset and better motor function and clinical outcome, and there was a significant association between SMN2 gene copy number and clinical outcome (P<0.05). CONCLUSIONS The SMN2 gene can reduce the severity of SMA via the dosage compensation effect. SMN2 copy number is associated with the phenotype of SMA, and therefore, it can be used to predict disease severity.
Child ; Humans ; Muscular Atrophy, Spinal ; genetics ; Phenotype ; Survival of Motor Neuron 1 Protein ; genetics ; Survival of Motor Neuron 2 Protein ; genetics

OBJECTIVE: To explore the clinical value of non-invasive prenatal testing(NIPT)for screening fetal chromosomal copy number variations(CNVs) and microdeletion/microduplication syndromes(MDs).METHODS: Retrospective analysis was made in the 10 005 women who received NIPT during the first trimester(15-20+ 6 weeks)from January,2012 to July,2017,at First People's Hospital of Yunnan Province,Department of Genetic Diagnosis Center.Among them 32 pregnant women were indicated fetal CNVs,25 of 32 pregnant women selected interventional prenatal diagnosis.Statistical analysis was made on the amniotic fluid/cord blood chromosome G band karyotype and high-throughput sequencing(NGS)genome copy number analysis was made,and relevant CNVs were searched and analyzed in the corresponding database;the consistency of CNVs found in NIPT with interventional prenatal diagnosis was statistically analyzed.RESULTS: During the second trimester(15-20+ 6 weeks),in the 10 005 pregnant women who received NIPT testing 32 cases were shown to have high risks of fetal CNVs,and the screening positive rate was 0.32%(32/10 005).In 25 high risk pregnant women who accepted invasive prenatal diagnosis via informed choice,14 women wereconfirmed as fetal CNVs,the positive predictive value(PPV)of NIPT being 56%(14/25),including 9 cases of microdeletion and 5 cases of microduplication.The sizes were between 587.75 kb and 36.05 Mb.The size and the start and end positions of CNVs found by NIPT were similar to those of fetal DNA samples detected by NGS.Among 14 cases of fetal CNVs,11 cases were identified as MDs,3 cases as unknown clinical significance.In 11 cases of MDs,8 cases were observed fetal chromosome structure abnormalities by karyotype analysis,10 cases were confirmed as de novo abbreviations,and 2 cases as originated from paternal same MD.After genetic counseling,10 pregnant women in 11 cases of MDs chose informed terminations,and one case chose continuing pregnancy.CONCLUSION: As a high-precision screening method,NIPT is expected to be an effective mean to screen for fetal CNVs,which can be used to detect highrisk chromosome microdeletion and microduplication CNVs of larger segments.High risk cases of fetal CNVs found by NIPT require invasive prenatal diagnosis for validation.

Objective To investigate the learning curve of contrast-enhanced ultrasonography ( CEUS) in sentinel lymph node( SLN ) of breast cancer and provide a theoretical basis for leaners to learn SLN CEUS . Methods The multi-center study of SLN CEUS in breast cancer" was planned by Sichuan Cancer Hospital . According to the uniform inclusion and exclusion criteria , 511 patients with complete clinical data and follow-up results from 9 hospitals in Multi-center were included in this study . According to the inspection time ,the patients were divided into 3 groups named as group A ( 170 patients) ,group B ( 170 patients) and group C ( 171 patients ) ,respectively . The basic clinical data ,ultrasound imaging data , intraoperative and postoperative pathological findings of all patients were recorded . With the accumulation of cases examined ,analysis was performed to find the learning curve of the SLN CEUS examination time , SLN CEUS detection rate ,SLN CEUS surface marking accuracy rate and SLN CEUS diagnosis rate ,the learning curve was analyzed . Results ① There was no statistical significant difference in patients ages , tumors sizes ,tumors locations ,SLNs numbers and LCs numbers among the three groups( all P > 0 .05) . ②As the number of cases examined increases ,the examination time was reduced gradually ,but SLN detection rate ,surface marking accuracy and SLN diagnostic coincidence rate were increased gradually( F = 151 .75 , 1 .96 ,7 .49 ,5 .50 ; P = 0 .000 ,0 .143 ,0 .001 ,0 .005 ) . Conclusions The skill of the doctor is improved gradually when learning SLN CEUS . With the number of the cases increase ,the operating time of SLN CEUS is shorted ,and the SLN detection rate ,surface marking accuracy and SLN diagnostic coincidence rate of SLN-CEUS are gradually increased . It has an important clinical significance for beginners to learn the SLN CEUS technology .

ObjectiveThe mechanism of paraquat poisoning (PQP) inducing acute lung injury is not clear. Nuclear factor erythroid 2-related factor 2 (Nrf2 )-antioxidant response element (ARE) is found to be a most important endogenous antioxidant defense pathway. This study aimed to explore the protective effect of 5- amino salicylic acid (5-ASA) against PQP-induced acute lung injury by activating the Nrf2-ARE pathway.MethodsEighty healthy adult male SD rats were randomly divided into four groups: blank control, 5-ASA control, PQP, and 5-ASA treatment. The animals in the PQP and 5-ASA treatment groups were injected with paraquat at 20 mg/kg into the left abdominal cavity for construction of the PQP model and those in the blank and 5-ASA control groups with isotonic saline at 1 mL. At 2 hours after modeling, the rats in the 5-ASA control and 5-ASA treatment groups received gavage of 5-ASA 75 at mg/kg for 3 successive days. At 1 and 3 days after observation, all the rats were sacrificed and the lower lobe of the right lung harvested for HE staining and observation of pathologic changes in the lung tissue. Meanwhile the left lung tissue was collected for determination of the expressions of Nrf2 and HO-1 proteins by Western blot.ResultsBehavioral changes were observed in the rats of the PQP and 5-ASA treatment groups, but less obvious in the latter. The alveolar wall capillaries of the rats in the PQP group were expanded and congested significantly, with widened alveolar septa and infiltration of a large number of inflammatory cells at 1 and 3 days, even severer at 3 days. The rats of the 5-ASA treatment group showed obviously reduced edema and the inflammatory cell infiltration at the corresponding time points as compared the PQP group. The lung tissue pathology scores were significantly higher in the 5-ASA treatment and PQP groups than in the blank control at 1 day (0.66±0.10 and 0.61±0.04 vs 0.18±0.05, P<0.05) and at 3 days (0.74±0.08 and 0.49±0.08 vs 0.16±0.02, P<0.05), but markedly lower in the 5-ASA treatment than in the PQP group (P<0.05). Both the expressions of Nrf2 and HO-1 proteins were remarkably higher in the 5-ASA control, PQP and 5-ASA treatment groups than in the blank control at 1 and 3 days (P<0.05), and so were they in the 5-ASA treatment than in the PQP group (P<0.05).Conclusion5-ASA can effectively reduce PQP-induced acute lung injury, which may be related to its up-regulation of the Nrf2 expression.

OBJECTIVETo study the clinical characteristics of urticaria in children versus adults, and to provide reference for the etiological analysis, disease evaluation, and treatment of urticaria in children.

METHODSThe clinical data of 2 411 patients with urticaria who visited the Department of Dermatology at Xiangya Hospital of Central South University from January 2013 to May 2017 were collected to study their socio-demographic characteristics. The clinical characteristics of urticaria were compared between the 68 children and 672 adults of the 740 patients with complete follow-up data.

RESULTSAmong the 411 pediatric patients, 314 (76.4%) had acute urticaria; among the 2 000 adult patients, 896 (44.8%) had chronic spontaneous urticaria. The causes of acute urticaria in children included infection (41%, 16/39). The accompanying symptoms of acute urticaria in children mainly included abdominal pain and diarrhea (44%, 17/39), while those in adults mainly included chest distress and shortness of breath (32%, 11/34). Compared with the adult patients, the pediatric patients had significantly lower chronic urticaria activity scores before and after treatment (P<0.05), a significantly higher rate of response to second-generation antihistamines (82.1% vs 62.2%; P<0.05), and a significantly higher proportion of individuals with a personal and family history of urticaria (P<0.05).

CONCLUSIONSAcute urticaria is more commonly seen than chronic urticaria in children with urticaria, and the main accompanying symptoms are abdominal pain and diarrhea, which are different from adults with urticaria. Chronic urticaria has a better treatment outcome in children than in adults. The most frequently seen cause of acute urticaria is infection in children. Atopic children may be susceptible to urticaria.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Urticaria ; drug therapy ; etiology ; Young Adult

Objective To investigate gene polymorphisms of drug targets and mutations associated with drug resistance in Pneumocystis jiroveci (P.jiroveci) isolates.Methods Among 148 samples isolated from human immunodeficiency virus (HIV)infected patients with pneumonia in Guangdong,mitochondrid larg subunit rRNA (mtLSUrRNA) gene was amplified from 51 samples.Dihydropteroate synthase (DHPS),dihydrofolate reductase (DHFR) and Cytochrome b (CYB) genes of P.jiroveci were detected by gene sequencing,and compared with the reference sequences in GenBank to evaluate gene polymorphisms.Results P.jirovecii DHPS,DHFR and CYB genes were all successfully amplified from 51 samples.For DHPS gene,48 (94.1％) were wild-type and 3 (5.9％) had gene mutation associated with drug resistance.For DHFR gene,30 were wild-type,and 21 had a synonymous mutation at position 312,and 1 nonsynonymous mutation at position 188.There were no mutations associated with drug resistance.For CYB gene,polymorphisms of were detected at 5 sites,4 of which were synonymous mutations,1 was non-synonymous mutation.No mutation associated with drug resistance was found.Based on the gene polymorphism of CYB6,the strains can be classified into 6 genotypes,and 2 were first detected,including 25 CYB1,13 CYB2,2 CYB5,4 CYB8,as well as newly detected 4 CYB10 and 3 CYB11 strains.Conclusions The mutations associated with drug resistance in P.jiroveci isolates in Guangdong remain uncommon.CYB gene shows gene polymorphisms and can be selected as one of targeted genes for multilocus sequence typing.