Five new flavan-4-ol glycosides jixueqiosides A-E (1-5) and two new flavan glycosides jixueqiosides F and G (6 and 7), along with twelve known flavan-4-ol glycosides (8-19), were isolated from the roots of Pronephrium penangianum. Comprehensive spectral analyses, X-ray single-crystal diffraction, and theoretical electronic circular dichroism (ECD) calculations established structures and absolute configurations. A single crystal structure of flavan-4-ol glycoside (14) was reported for the first time, while the characteristic ECD and NMR data for all isolated flavan-4-ol glycosides (1-5 , 8-19) were analyzed, establishing a set of empirical rules. Activity screening of these isolates showed that 8 and 9 could inhibit the proliferation of MDA-MB-231 and MCF-7 cells with IC₅₀ values of 7.93 ? 2.85 ?mol?L^-1 and 5.87 ? 1.58 ?mol?L^-1 (MDA-MB-231), and 2.21 ? 1.38 ?mol?L^-1 and 3.52 ? 1.55 ?mol?L^-1 (MCF-7), respectively. Western blotting and flow cytometry analyses demonstrated that 8 and 9 dose-dependently induced apoptosis in MDA-MB-231 cells by up-regulating BAX, activating caspase-3 and down-regulating BCL-2. Additionally, compound 8 affected autophagy-related proteins, increasing the ratio of LC3-II/LC3-I and Beclin-1 levels to inhibit MDA-MB-231 cell proliferation. Moreover, anti-inflammatory studies indicated that 2, 3, 7, 13, 14, and 18 moderately inhibited tumor necrosis factor-a (TNF-a), interleukin-6 (IL-6), and nitric oxide (NO) release.
Humans ; Plant Roots/chemistry* ; Glycosides/isolation & purification* ; Anti-Inflammatory Agents/isolation & purification* ; Flavonoids/isolation & purification* ; Cell Proliferation/drug effects* ; Antineoplastic Agents, Phytogenic/isolation & purification* ; Molecular Structure ; Apoptosis/drug effects* ; Cell Line, Tumor ; Tumor Necrosis Factor-alpha/immunology* ; Drugs, Chinese Herbal/pharmacology* ; Interleukin-6/immunology* ; Animals ; Mice

OBJECTIVE: To explore the clinical and genetic characteristics of a patient with Hermansky-Pudlak syndrome type 5 (HPS-5). METHODS: A child with HPS-5 who had attended the Children's Hospital Affiliated to Shandong University on October 3, 2019 was selected as the study subject. Clinical data of the child were collected. Genetic variant was analyzed through high-throughput sequencing. A literature review was also carried out. RESULTS: The child, a 1-year-and-5-month-old girl, had nystagmus since childhood, lost of retinal pigmentation by fundus examination and easy bruising. High-throughput sequencing revealed that she has harbored compound heterozygous variants of the HPS5 gene, namely c.1562_1563delAA (p.F521Sfs*27) and c.1404C>A (p.C468X), which were inherited from his father and mother, respectively. Based on the guidelines from the American College for Medical Genetics and Genomics (ACMG), both variants were predicted to be pathogenic (PVS+PM2_Supporting+PM3+PP4). Among 18 previously reported HPS-5 patients, all had had eye problems, and most of them had tendency for bleeding. Eight cases had carried compound heterozygous variants of the HPS5 gene, 8 carried homozygous variants, 2 carried double homozygous variants, and most of them were null mutations. CONCLUSION The c.1562_1563delAA(p.F521Sfs*27) and c.1404C>A (p.C468X) compound heterozygous variants of the HPS5 gene probably underlay the HPS-5 in this child. High-throughput sequencing has provided an important tool for the diagnosis. HSP-5 patients usually have typical ocular albinism and/or oculocutaneous albinism and tendency of bleeding, which are commonly caused by compound heterozygous and homozygous variants of the HPS5 gene, though serious complications have been rare.
Female ; Humans ; Infant ; Hermanski-Pudlak Syndrome/pathology* ; High-Throughput Nucleotide Sequencing ; Mutation

OBJECTIVE: To explore the clinical feature and genetic etiology of a patient with normosmic idiopathic hypogonadotropic hypogonadism (nIHH) due to variant of CHD7 gene. METHODS: A patient who had presented at Anhui Provincial Children's Hospital in October 2022 was selected as the study subject. Clinical data of the patient was collected. The patient and his parents were subjected to trio-whole exome sequencing. Candidate variant was verified by Sanger sequencing and bioinformatic analysis. RESULTS: The patient had featured delayed development of secondary sexual characteristics but normal olfactory function. Genetic testing revealed that he has harbored a c.3052C>T (p.Pro1018Ser) missense variant of the CHD7 gene, for which both of his parents were of the wild type. The variant has not been recorded in the PubMed and HGMD databases. Analysis of amino acid sequences suggested that the variant site is highly conserved, and the variant may affect the stability of protein structure. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.3032C>T variant was classified as a likely pathogenic (PS2+PM2_Supporting+PP2+PP3+PP4). CONCLUSION The delayed development of secondary sexual characteristics of the patient may be attributed to the c.3052C>T (p.Pro1018Ser) variant of the CHD7 gene. Above finding has expanded the variation spectrum of the CHD7 gene.
Child ; Humans ; Male ; Amino Acid Sequence ; Computational Biology ; DNA Helicases/genetics* ; DNA-Binding Proteins/genetics* ; Genetic Testing ; Genomics ; Hypogonadism/genetics* ; Mutation

OBJECTIVE: To investigate the clinical characteristics and genetic basis for a child with Keppen-Lubinsky syndrome (KPLBS). METHODS: Trio-whole exome sequencing (Trio-WES) was carried out for the proband and her parents. Candidate variant was verified by Sanger sequencing and bioinformatic analysis. RESULTS: The child has featured peculiar facies including large eyes, alar hypoplasia, microretrognathia, premature aging appearance in addition with growth delay and mental retardation. Trio-WES has identified that she has carried a de novo variant of the KCNJ6 gene, namely c.460G>C (p.Gly154Arg). The variant has not been recorded in the database. Prediction of protein structure indicated that the variant may affect the potassium ion selective filtration structure channel in the transmembrane region of KCNJ6 protein, which may result in up regulation of the function of the channel. CONCLUSION The de novo c.460G>C (p.Gly154Arg) variant of the KCNJ6 gene probably underlay the KPLBS in this child. Above finding has enriched the genotypic and phenotype spectrum of this syndrome.
Cataract ; China ; Female ; G Protein-Coupled Inwardly-Rectifying Potassium Channels/genetics* ; Humans ; Hypogonadism/congenital* ; Intellectual Disability/genetics* ; Mutation ; Whole Exome Sequencing

Conjunctival melanoma (CM) is a rare and fatal malignant eye tumor. In this study, we deciphered a novel anti-CM mechanism of a natural tetracyclic compound named as cucurbitacin B (CuB). We found that CuB remarkably inhibited the proliferation of CM cells including CM-AS16, CRMM1, CRMM2 and CM2005.1, without toxicity to normal cells. CuB can also induce CM cells G2/M cell cycle arrest. RNA-seq screening identified KIF20A, a key downstream effector of FOXM1 pathway, was abolished by CuB treatment. Further target identification by activity-based protein profiling chemoproteomic approach revealed that GRP78 is a potential target of CuB. Several lines of evidence demonstrated that CuB interacted with GRP78 and bound with a K _d value of 0.11 μmol/L. Furthermore, ATPase activity evaluation showed that CuB suppressed GRP78 both in human recombinant GRP78 protein and cellular lysates. Knockdown of the GRP78 gene significantly induced the downregulation of FOXM1 and related pathway proteins including KIF20A, underlying an interesting therapeutic perspective. Finally, CuB significantly inhibited tumor progression in NCG mice without causing obvious side effects in vivo. Taken together, our current work proved that GRP78-FOXM1-KIF20A as a promising pathway for CM therapy, and the traditional medicine CuB as a candidate drug to hinder this pathway.

Objective:To investigate the role of miR-146a in the pathogenesis of systemic juvenile idiopathic arthritis (sJIA) and its clinical significance.Methods:This article is a prospective clinical cohort study.Twenty-six patients with sJIA (14 cases of initial active group and 12 cases of stable group), 15 patients with multijoint juvenile idiopathic arthritis (JIA) and 15 patients with oligojoint JIA diagnosed in the Department of Rheumatology and Immunology of Anhui Provincial Children′s Hospital from June 2018 to December 2020 were enrolled.Twenty healthy controls from the out-patient clinic were also recruited.The expression level of miR-146a in peripheral blood mononuclear cells (PBMCs) of research objects was detected by real-time fluorescence quantitative polymerase reaction (qPCR), and the serum levels of interleukin (IL) - 6, tumor necrosis factor (TNF) - α and IL-1β in sJIA patients and healthy controls were detected by enzyme-linked immunosorbent assay.The expression levels of miR-146a in PBMCs and cytokines among different groups were compared by analysis of variance. Pearson correlation analysis was used to analyze the correlation of the relative expression level of miR-146a in PBMCs with clinical inflammatory indexes and serum cytokines in sJIA patients. Results:(1) The expression level of miR-146a in PBMCs of early sJIA patients was significantly higher than that in the multijoint JIA group and oligojoint JIA group (8.77±3.15 vs.4.40±1.59, 2.55±1.15, t=6.27, 14.23; all P<0.05). The expression level of miR-146a in PBMCs of sJIA active patients was significantly higher than that in sJIA stable patients (8.77±3.15 vs.3.63±1.37, t=10.27, P<0.05). There was no significant difference in the expression level of miR-146a between the sJIA stable group and healthy control group ( P>0.05). (2) The expression levels of IL-1β, IL-6 and TNF-α were significantly higher in sJIA active patients group than those in sJIA stable group[(58.56±17.47) ng/L vs.(26.32±10.54) ng/L, (73.72±11.16) ng/L vs.(23.20±9.12) ng/L, (70.93±19.97) ng/L vs.(24.25±9.49) ng/L, all P<0.05]. There was no significant difference in the expression levels of IL-1β, IL-6 and TNF-α between the sJIA stable group and healthy control group(all P>0.05). (3)The expression of miR-146a in PBMCs of sJIA patients was positively correlated with serum ferritin levels, platelets, erythrocyte sedimentation rates, C-reactive proteins, IL-1β, IL-6 and TNF-α( r=0.542, 0.433, 0.329, 0.306, 0.333, 0.342, 0.319, all P<0.05). Conclusions:miR-146a may be involved in the inflammatory process of sJIA disease.miR-146a can well distinguish sJIA from multijoint JIA and oligojoint JIA.TNF-α, IL-1β and IL-6 are involved in sJIA inflammatory responses.

Objective:To evaluate the feasibility, safety and efficacy of single-stage percutaneous nephrolithotomy (PCNL) combined with flexible ureteroscopy (FURS) for the management of staghorn calculi with pyonephrosis.Methods:A total of 13 patients of staghorn renal calculi with pyonephrosis, which was diagnosed by intraoperative pelvic urine bacteria culture, were treated by PCNL combined with FURS from May 2017 to December 2019. Of all the 13 patients, 7 were males and 6 were females, with mean age of 52.5 years, ranged from 33 years to 68 years. The mean stone burden was (1 070.9±397.0) mm 2, ranged from 507.4 mm 2 to 1 809.5 mm 2. Bacteria culture and microbial sensitivity test was performed for all the patients. Four patients with fever on admission accepted ureteral stenting at least a week before the surgeries. All the patients received preoperative antibiotic therapies for at least a week, and the infective symptoms and inflammatory indexes was normal before the surgeries. Under general anesthesia, the procedures were performed in a modified supine Valdivia position. After the transurethral FURS was performed, the standard percutaneous track was placed at the subcostal point between mid-axillary line and scapular line under the FURS and ultrasounic guide. The purulence and the stones were shattered and removed by PCNL with negative pressure system, and FURS helped to shatter and move out the stones beyond the PCNL reach through the same tract. A double-J tubes and a nephrostomy tube was routinely indwelled postoperatively. A radiological imaging was performed within three days after the operation to evaluate the stone free rate. The residual stone was defined as the stone larger than 4 mm. Results:All the procedures were successful. The one-stage stone free rate was 76.9% (10/13). The mean operation time was (94.2±21.8) min, ranged from 65 to 135 min. Six patients suffered postoperative systemic inflammatory response syndrome, and no patient occurred sepsis or complications of Clavien-Dindo classification Ⅲ or above. They were followed up for 3-24 months with median of 12 months.Four cases with recurrence of ipsilateral stones, two cases with ipsilateral mild renal atrophy, no recurrent ipsilateral upper uninary infection were found in the followup.Conclusions:Single-stage PCNL combined with flexible ureteroscopy could be feasible for the management of staghorn calculi with pyonephrosis with safety and efficacy.

Objective To explore the genetic basis for a neonate featuring global developmental delay.Methods Clinical and laboratory tests were carried out for the patient.Peripheral venous blood samples were collected from the neonate and his parents for the extraction of DNA.Potential variant was detected by using targeted capture and next generation sequencing for a panel of genes associated with nervous system diseases.Suspected variant was validated by Sanger sequencing.Results The nine-month-old boy manifested global developmental delay and was unstable to sit alone and distinguish strangers from acquaintance.Genetic testing revealed two novel variants of the SLC19A3 gene in him,namely c.448G＞ A and c.169C＞T.The amino acids encoded by the two codons are highly conservative,and both variants were predicted to be pathogenic by bioinformatic analysis.Conclusion The compound heterozygous c.448G＞A and c.169C＞T variants probably underlay the onset of disease in the patient.Above finding also enriched the variant spectrum of SLC19A3 gene underlying Biotin-thiamine responsive basal ganglia disease.

Objective To describe the epidemiological characteristics of cervical spine fracture combined with trauma cervical spinal cord injury (TCSCI) in Guangxi province. Methods A total of 385 patients met the inclusion criteria were enrolled for retrospective investigation. Detailed information included age, sex, marital status, occupation, date of admission, mechanisms of trauma, level of injury, ASIA grade, concomitant injuries, length of hospital stay, complications, and mortality. Results The ratio of Male/Female was 4.4:1. The average age of the patients was 47.9 years, and 88.8％ of the patients were married. 72.9％ of TCSCI occurred between the age of 35-64 years. Farmers accounted for the largest number of patients with TCSCIs, and 63％ of patients with TCSCI were caused by falling. The damage was located at the C3-C5 level, accounting for 63.8％. More than half of the patients with CSCI had brain injury. The most common complication was respiratory infection (54.5％). Among the injury levels, the proportions of ASIA grade A, B, C, and D were 34.8％, 8.5％, 35.5％, and 21％, respectively. During the treatment, 58 patients required cardiopulmonary resuscitation, and 36 patients required mechanical ventilation. The average length of hospital stay was (26.5±21.6) d. Of the 375 patients discharged from the hospital, 51.2％ patients had no improvement in the muscle strength. Conclusions Falling is the main cause of CSCI and men are more likely to be exposed to the injury. Patients with TCSCI have long treatment time and poor treatment results. Additionally, complications during the treatment should not be ignored.

Objective: To investigate application value of "point-line-surface-volume" four dimensional holmium laser enucleation of prostate (HoLEP) in the treatment of benign prostatic hyperplasia (BPH). Methods: From December 2016 to November 2017, the clinical data of 60 cases of BPH with the treatment of "Point-line-surface-volume" four dimensional HoLEP at Department of Urology, Peking University Third Hospital were analyzed retrospectively. The age was (73.8±8.0) years (range: 60 to 96 years), body mass index was (23.2±3.6) kg/m² (range: 14.9 to 31.1 kg/m²), volume of prostate was (64.5±36.9) ml (range: 15.5 to 197.9 ml). All the cases were operated by the same chief surgeon expert in endoscopic surgery. First of all, taking verumontanum as the anchor point, crossing point between the outer 45° line of verumontanum and middle line of the lateral lobe and verumontanum was made as the breakthrough point. The "blasting and paddling" method was used to find prostatic surgery capsule, and the two sides of the capsule plane were connected in front of the verumontanum. Secondly, the plane was maintained by the "fan tunnel" method, and the middle lobe and the lateral lobes were divided at the 5 o′clock and 7 o′clock positions. The glands of the middle lobe were first removed and pushed into the bladder. Then, on the anteroinferior inclined coronal plane through 12 o′clock point right above of verumontanum, the urethral mucosa was cut apart in a semi-curved shape in advance. Bilateral semi-curved incision confluenced above the verumontanum, and extend to the bladder neck to form an inverted Y-shaped groove which completely divided the bilateral lobes. Finally, the bilateral lobes were enucleated respectively, and glands in the bladder were removed by tissue morcellator. Clinical data included operative time, hemoglobin decrease, catheterization duration, postoperative hospital stay, preoperative and postoperative international prostate symptom score and quality of life (QOL) score. The efficiency of enucleation was calculated as prostatic volume divided by enucleation time (not including morcellation time). The efficiency of morcellation was calculated as prostatic volume divided by morcellation time. Paired t-test was used to compare the indexes before and after surgery. Results: All the operations of 60 cases were sumlessful, one of which prostate tissue was removed by conversion to transurethral resection prostate due to malfunction of tissue morcellator. The operative time was (115.2±52.9) minutes (range: 25 to 276 minutes). The enucleation efficiency was (0.81±0.35) ml/minutes (range: 0.17 to 1.58 ml/minutes). The morcellation efficiency was (6.60±4.28) ml/minutes (range: 0.89 to 17.42 ml/minutes). The hemoglobin was decreased by (15.9±12.3) g/L (range:-10 to 57 g/L). Meanwhile, catheterization duration was (5.2±2.9) days (range: 0.8 to 19.8 days), and postoperative hospital stays were (5.0±1.5) days (range: 1.9 to 11.9 days). Preoperative and postoperative IPSS scores (10.92±6.98 vs. 23.37±7.49, t=10.357, P=0.000) and QOL scores (1.75±1.62 vs. 4.53±1.47, t=9.373, P=0.000) were significantly different. Postoperative complications included: 4 cases of fever (greater than 38.5 ℃), 1 case of acute epididymitis, 1 case of bladder mucosal injury, and 1 case of active bleeding need blood transfusion therapy. Conclusion "Point-line-surface-volume" four dimensional HoLEP is an effective and safe minimally invasive method in the treatment of BPH.