A Single Set of Well-Designed Aptamer Probes for Reliable On-site Qualitative and Ultra-Sensitive Quantitative Detection.

Aptamer-based probes are pivotal components in various sensing strategies, owing to their exceptional specificity and versatile programmable structure. Nevertheless, numerous aptamer-based probes usually offer only a single function, limiting their capacity to meet the diverse requirements of multi-faceted sensing systems. Here, we introduced supersandwich DNA probes (SSW-DNA), designed and modified on the outer surface of nanochannels with hydrophobic inner walls, enabling dual functionality: qualitative detection for on-site analysis and quantitative detection for precise analysis. The fragmented DNAs resulting from the target recognition, are subsequently identified through lateral flow assays, enabling robust on-site qualitative detection of microcystin-LR with an impressively low limit of detection (LOD) at 0.01 µg/L. Meanwhile, the nanochannels enable highly sensitive quantification of microcystin-LR through the current analysis, achieving an exceptionally low LOD at 2.5×10-7  µg/L, with a broad dynamic range spanning from 1×10-6 to 1×102  µg/L. Furthermore, the process of target recognition introduces just a single potential error propagation, which reduces the overall risk of errors during the entire qualitative and quantitative detection process. This sensing strategy broadens the scope of applications for aptamer-based composite probes, holding promising implications across diverse fields, such as medical diagnosis, food safety, and environmental protection.

Inhibiting von Hippel‒Lindau protein-mediated Dishevelled ubiquitination protects against experimental parkinsonism.

Dopaminergic neuron degeneration is a hallmark of Parkinson's disease (PD). We previously reported that the inactivation of von Hippel‒Lindau (VHL) alleviated dopaminergic neuron degeneration in a C. elegans model. In this study, we investigated the specific effects of VHL loss and the underlying mechanisms in mammalian PD models. For in vivo genetic inhibition of VHL, AAV-Vhl-shRNA was injected into mouse lateral ventricles. Thirty days later, the mice received MPTP for 5 days to induce PD. Behavioral experiments were conducted on D1, D3, D7, D14 and D21 after the last injection, and the mice were sacrificed on D22. We showed that knockdown of VHL in mice significantly alleviated PD-like syndromes detected in behavioral and biochemical assays. Inhibiting VHL exerted similar protective effects in MPP+-treated differentiated SH-SY5Y cells and the MPP+-induced C. elegans PD model. We further demonstrated that VHL loss-induced protection against experimental parkinsonism was independent of hypoxia-inducible factor and identified the Dishevelled-2 (DVL-2)/ß-catenin axis as the target of VHL, which was evolutionarily conserved in both C. elegans and mammals. Inhibiting the function of VHL promoted the stability of ß-catenin by reducing the ubiquitination and degradation of DVL-2. Thus, in vivo overexpression of DVL-2, mimicking VHL inactivation, protected against PD. We designed a competing peptide, Tat-DDF-2, to inhibit the interaction between VHL and DVL-2, which exhibited pharmacological potential for protection against PD in vitro and in vivo. We propose the therapeutic potential of targeting the interaction between VHL and DVL-2, which may represent a strategy to alleviate neurodegeneration associated with PD.

A novel PPRC1 point mutation in a Chinese family with premature ovarian failure: A case study.

BACKGROUND: Patients with premature ovarian failure (POF) have an at least 6-month history of amenorrhea and elevated follicle-stimulating hormone levels in plasma. Most of the POF causes are idiopathic and hereditary, and chromosomal abnormalities have been associated with POF development. A pedigree study was performed on a family with idiopathic POF to observe the possible link between gene mutation and POF development. METHODS: In total, eight women were diagnosed with POF and seven POF patients and five non-POF members from the same family were evaluated by whole exome sequencing and Sanger sequencing. An apoptotic assay, senescence staining, real-time polymerase chain reaction (qPCR) and overexpression of the peroxisome proliferator-activated receptor gamma coactivator-related 1 (PPRC1) gene were performed to examine the association of POF in vitro. RESULTS: Through whole exome sequencing and Sanger sequencing, a novel point mutation (NM_015062: c.2902C>T:p.Thr958Ile) was identified and verified in the PPRC1 gene on chromosome 10 (10q24.32). The point mutation only presented in all the seven POF cases and not in non-POF cases or public databases. Subsequent expression of PPRC1 in COV434 granulosa cells showed that PPRC1 might be involved in regulating granulosa cell apoptosis but not senescence-associated POF development. CONCLUSIONS: A novel point mutation in the PPRC1 gene was identified by the pedigree study and by sequence analysis of the case series with idiopathic POF in the present study. The subsequent PPRC1 expression analysis showed that PPRC1 was not involved in senescence-associated POF development. Further studies will be needed to confirm the link between PPRC1 gene mutation and POF.

Design, synthesis, and initial evaluation of affinity-based small molecular probe for detection of WDR5.

WDR5, a subunit of the SET/MLL complex, plays critical roles in various biological progresses and are abnormally expressed in many cancers. Here we report the design, synthesis, and biochemical characterization of a new chemical tool to capture WDR5 protein. The probe is a biotinylated version of compound 30 that is a potent WDR5 inhibitor we previously reported. Importantly, the probe displayed high affinity to WDR5 protein in vitro binding potency and showed the ability in specifically and real time monitoring WDR5 protein. Further, the biotinylated tag of the probe enabled selectively "chemoprecipitation" of WDR5 from whole cell lysates of MV4-11. This probe provided a new approach to identify the overexpressed WDR5 protein in different cancer cells and applications to proteomic analysis of WDR5 and WDR5-binding partners.

Effect of hyaluronic acid-enriched transfer medium on frozen-thawed embryo transfer outcomes.

AIM: To determine if hyaluronic acid-enriched transfer medium (HETM) affects the implantation rate (IR) and clinical pregnancy rate (PR) in women undergoing frozen-thawed embryo transfer (FET). METHODS: The records of women who underwent FET from May 2014 to October 2014 were retrospectively reviewed. Outcome measures were IR and PR. RESULTS: In all 1721 cycles of 1632 patients were included in this study. HETM was used for 347 cycles of 342 patients, and standard medium for 1374 cycles of 1290 patients. Overall, FET outcomes were similar between the groups. For patients undergoing their first FET attempt, the IR (24.3% vs 31.6%, P = 0.042) and clinical PR (34.3% vs 50.1%, P = 0.004) were lower in the HETM group. For patients undergoing their second FET attempt, pregnancy outcomes were similar between the groups. For patients undergoing their third or more FET attempt, HETM was associated with a higher IR (33.3% vs 16.4%, P < 0.001) and higher PR (52.2% vs 27.4%, P < 0.001). CONCLUSIONS: HETM can improve the embryo IR and clinical PR in patients with repeated implantation failure in the third or more FET attempt. However, the use of HETM for first and second FET should be done with caution.

Discovery, design and synthesis of 6H-anthra[1,9-cd]isoxazol-6-one scaffold as G9a inhibitor through a combination of shape-based virtual screening and structure-based molecular modification.

Protein lysine methyltransferase G9a is widely considered as an appealing antineoplastic target. Herein we present an integrated workflow combining shape-based virtual screening and structure-based molecular modification for the identification of novel G9a inhibitors. The shape-based similarity screening through ROCS overlay on the basis of the structure of UNC0638 was performed to identify CPUY074001 contained a 6H-anthra[1,9-cd]isoxazol-6-one scaffold as a hit. Analysis of the binding mode of CPUY074001 with G9a and 3D-QSAR results, two series compounds were designed and synthesized. The derivatives were confirmed to be active by in vitro assay and the SAR was explored by docking stimulations. Besides, several analogues showed acceptable anti-proliferative effects against several cancer cell lines. Among them, CPUY074020 displayed potent dual G9a inhibitory activity and anti-proliferative activity. Furthermore, CPUY074020 induced cell apoptosis in a dose-dependent manner and displayed a significant decrease in dimethylation of H3K9. Simultaneously, CPUY074020 showed reasonable in vivo PK properties. Altogether, our workflow supplied a high efficient strategy in the identification of novel G9a inhibitors. Compounds reported here can serve as promising leads for further study.

The association of serum estradiol level with outcomes of clomiphene citrate/human menopausal gonadotropin ovarian stimulation for in vitro fertilization and embryo transfer.

BACKGROUND: The purpose of this study was to test the hypothesis that estradiol (E2) level on day 3 may be associated with in vitro fertilization (IVF) outcomes. METHODS: The records of patients who received clomiphene citrate 100 mg/day plus human menopausal gonadotropin 150 IU/day from day 3 and received frozen-thawed embryo transfers were reviewed. Patients were divided into three groups: group A (E2 ≤30 pg/ml), group B (30< E2 ≤50 pg/ml), and group C (E2 >50 pg/ml). A total of 1080 cycles from 941 patients were included. RESULTS: The number of eggs and MII oocytes were less in group C than group A (both, P = 0.001). The embryo implantation (P = 0.006) and clinical pregnancy rates (P = 0.036) were lower in group C than group B, and the rates were similar between group A and B. CONCLUSION: Maintaining the serum E2 level from 30 to 50 pg/ml may result in a higher clinical pregnancy rate in IVF cycles.

Investigation of the intermolecular recognition mechanism between the E3 ubiquitin ligase Keap1 and substrate based on multiple substrates analysis.

E3 ubiquitin ligases are attractive drug targets due to their specificity to the ubiquitin machinery. However, the development of E3 ligase inhibitors has proven challenging for the fact that they must disrupt protein-protein interactions (PPIs). The E3 ligase involved in interactome provide new hope for the discovery of the E3 ligase inhibitors. These currently known natural binding partners of the E3 ligase can benefit the discovery of other unknown substrates and also the E3 ligase inhibitors. Herein, we present a novel strategy that using multiple substrates to elucidate the molecular recognition mechanism of E3 ubiquitin ligase. Molecular dynamics simulation, molecular mechanics-generalized born surface area (MM-GBSA) binding energy calculation and energy decomposition scheme were incorporated to evaluate the quantitative contributions of sub-pocket and per-residue to binding. In this case, Kelch-like ECH-associated protein-1 (Keap1), a substrate adaptor component of the Cullin-RING ubiquitin ligases complex, is applied for the investigation of how it recognize its substrates, especially Nrf2, a master regulator of the antioxidant response. By analyzing multiple substrates binding determinants, we found that both the polar sub-pockets (P1 and P2) and the nonpolar sub-pockets (P4 and P5) of Keap1 can make remarkable contributions to intermolecular interactions. This finding stresses the requirement for substrates to interact with the polar and nonpolar sub-pockets simultaneously. The results discussed in this paper not only show the binding determinants of the Keap1 substrates but also provide valuable implications for both Keap1 substrate discovery and PPI inhibitor design.

[Progress in research of the structural optimization of natural product-like Garcinia caged xanthones].

Designing of natural product-like compounds using natural products as template structures is an important strategy for the discovery of new drugs. Gambogic acid (GA), which is a Garcinia natural product with a unique caged xanthone scaffold, inhibits potent antitumor activity both in vitro and in vivo. This review summarized the researches on the identification of the antitumor pharmacophore of GA, and the design, structural optimization and structure-activity relationship (SAR) of natural product-like caged xanthones based on it.

Synthesis and antibacterial evaluation of a novel series of 10-hydroxyl ketolide derivatives.

A novel series of 10-hydroxyl ketolide derivatives were synthesized, during which a distinctive intermediate, 3-O-descladinosyl-3-oxo-11-deoxy-10,11-epoxy-6-O-methylerythromycin A, was obtained from 6-O-methylerythromycin A. The structure and stereochemistry of this novel structure were confirmed via NMR and X-ray crystallography. Moreover, antibacterial evaluations were established in order to assess our modifications and acquire a deep understanding of the ketolides' structure-activity relationship (SAR).

Effective screening strategy using ensembled pharmacophore models combined with cascade docking: application to p53-MDM2 interaction inhibitors.

Protein-protein interactions (PPIs) play a crucial role in cellular function and form the backbone of almost all biochemical processes. In recent years, protein-protein interaction inhibitors (PPIIs) have represented a treasure trove of potential new drug targets. Unfortunately, there are few successful drugs of PPIIs on the market. Structure-based pharmacophore (SBP) combined with docking has been demonstrated as a useful Virtual Screening (VS) strategy in drug development projects. However, the combination of target complexity and poor binding affinity prediction has thwarted the application of this strategy in the discovery of PPIIs. Here we report an effective VS strategy on p53-MDM2 PPI. First, we built a SBP model based on p53-MDM2 complex cocrystal structures. The model was then simplified by using a Receptor-Ligand complex-based pharmacophore model considering the critical binding features between MDM2 and its small molecular inhibitors. Cascade docking was subsequently applied to improve the hit rate. Based on this strategy, we performed VS on NCI and SPECS databases and successfully discovered 6 novel compounds from 15 hits with the best, compound 1 (NSC 5359), K(i) = 180 ± 50 nM. These compounds can serve as lead compounds for further optimization.

Studies on chemical-structure modification and structure-activity relationship of gambogic acid derivatives at carbon(34).

Gambogic acid (GA), a natural product, was identified as a promising antitumor agent. To further explore the structure-activity relationship of GA and discover novel GA derivatives as antitumor agents, 19 novel GA derivatives modified at C(34) were synthesized and evaluated against A549, BGC-823, U251, HepG2, and MB-231 cancer cell lines by cellular assays. Among them, 15 compounds were found to be more potent than GA against some cancer cell lines. Notably, compound 3 possessed potent inhibitory activities against five cell lines with IC(50) values ranging between 0.24 and 1.09 µM. Compounds 9 and 18 were seven to eightfold more active than GA against A549 cell line. Chemical modification at C(34) of GA by introducing of hydrophilic aliphatic amines resulted in increased activity and improved drug-like properties. These findings will enhance our understanding of the SAR of GA and can lead to the discovery of novel GA derivatives as potential antitumor agents.

The Relationship Between Upper Esophageal Sphincter Pressure and Psychological Status in Patients with Globus Sensation.

OBJECTIVE: To explore the correlation between changes in esophageal pressure and psychological status in patients with globus sensation. METHODS: A total of 40 patients with globus sensation who attended Wenzhou People's Hospital between August 2020 and February 2021 were divided into two groups based on the results of esophageal manometry: a high-pressure group and a non-high-pressure group. The duration of disease, clinical symptom score, and self-rating anxiety scale (SAS) were compared between the two groups to determine the relationship between changes in esophageal pressure and psychological status. RESULTS: All the patients before treatment were divided into a high-pressure group (n = 14) and a non-high-pressure group (n = 26) according to whether the resting pressure of the upper esophageal sphincter (UES) was greater than 104 mmHg. The differences between the high-pressure group and non-high-pressure group in duration of disease, clinical symptom score, and SAS were statistically significant (all P < 0.05). Anxiety was present in 12 patients in the high-pressure group and two patients in the non-high-pressure group. The difference between the the high-pressure group and non-high-pressure group in the incidence of anxiety was statistically significant (χ2 = 21.04 and P < 0.001). Pearson correlation analysis of the association between esophageal pressure and anxiety resulted in R = 0.74 and P < 0.001. CONCLUSION: Patients with globus sensation who develop anxiety were more likely to have high pressure in the upper esophageal sphincter.

Fist-Edge-Palm (FEP) test has a high sensitivity in differentiating dementia from normal cognition in Parkinson's disease.

BACKGROUND: The Fist-Edge-Palm (FEP) test takes 0.5-3 min to complete and is highly sensitive in differentiating Alzheimer's disease and frontotemporal dementia from normal cognition, but it has not yet been studied in Parkinson's disease (PD). OBJECTIVE: To determine the sensitivity and specificity of the FEP test in screening patients with PD for cognitive impairment and dementia. METHODS: PD patients were recruited and divided into three groups based on cognitive status: normal cognition, mild cognitive impairment (MCI) and dementia according to 2015 MDS clinical diagnostic criteria for PD and clinical dementia rating scale (CDR) assessment for cognitive status. MMSE, FEP and clock drawing test (CDT) were tested in all recruited PD patients. Chi-square test was used to compare the sensitivity of FEP and CDT in detecting PDD and PD-MCI. RESULTS: A total of 108 PD patients were included: 52 normal cognition, 28 MCI, and 28 dementia. The sensitivity of FEP in differentiating PDD from PD-NC was 96.4% and the sensitivity for PD-MCI from PD-NC was 71.4%. The sensitivity of CDT in differentiating PDD from PD-NC was 71.4% and PD-MCI from PD-NC was 53.6%. The sensitivities of FEP and CDT were 83.9% and 62.5%, respectively, in identifying cognitive impairment (CDR ≥ 0.5) in PD patients. CONCLUSION: FEP is a sensitive screening tool in differentiating PDD or PD-MCI from PD-NC, and it is much faster than MMSE and more sensitive than CDT. FEP may be a practical screening tool for daily clinical practice.

Comparatively lower postoperative hepatolithiasis risk with hepaticocholedochostomy versus hepaticojejunostomy.

BACKGROUND: Optimal surgical technique plays a key role in preventing the postoperative recurrence of hepatolithiasis. Tian et al developed the subcutaneous tunnel and hepatocholangioplasty using the gallbladder (STHG) technique and applied it in hepatolithiasis patients who had an approximately normal gallbladder and sphincter of Oddi. However, the technique is controversial. In the present study, a canine model was established for hepatocholangioplasty (HC) and hepaticojejunostomy (HJ) to simulate STHG and Roux-en-Y cholangiojejunostomy in the clinic, respectively. Then, the alterations of bile components in the vicinity of the anastomosis were compared. This may provide an experimental guide for choosing an optimal technique for the treatment of hepatolithiasis in the clinic. METHODS: The animals were randomly separated into a control group (5 dogs) and a model group (stenosis of the common bile duct; 24 dogs). The 24 dogs in the model group were randomly divided into an HC group and an HJ group (12/group). Bile was collected from the bile duct at 1 and 5 months after the operation, and the bile components were determined. RESULTS: The levels of total bile acid, cholesterol, total bilirubin, and phospholipid in the HC group were higher than those in the HJ group (P<0.05). However, no statistical difference was seen in unconjugated bilirubin and calcium ions. The mucin level in bile in the HC group was lower than that in the HJ group at 5 months after the operation (P<0.05). The postoperative lipid peroxidation level was remarkably lower than that in the HJ group (P<0.05). However, the superoxide dismutase level was remarkably higher than that in the HJ group (P<0.05). Finally, a significant difference was found in the positive bacterial culture rate in bile between the groups. CONCLUSION: Changes of bile components near the anastomosis after HC might be more preferable for preventing hepatolithiasis formation than HJ.

[Prelimary exploration on Chinese medicine syndrome type distribution in patients with polycystic ovary syndrome].

OBJECTIVE: To explore the Chinese medicine syndrome type distribution in patients with polycystic ovary syndrome (PCOS) and its relationship with sexual hormones. METHODS: Chinese medicine syndrome types of 212 PCOS patients were differentiated and sorted by adopting fuzzy mean C clustering method, and their relationship with the indices of sexual hormones detected on the 3rd to 5th day of menstrual cycle was analyzed, with the values got from 20 healthy women for controls. RESULTS: Intermingling syndromes were commonly seen in PCOS patients. Shen-deficiency syndrome (presented in 64 patients) and Gan-qi stagnancy syndrome (61 patients) were the dominance, accounting for 30.2% and 28.8% respectively, significantly higher than that of other syndromes (P < 0.05), which were Pi-deficiency syndrome (41 patients, 19.3%), phlegm-dampness syndrome (33 patients, 15.6%) and blood stasis syndrome (13 patients, 6.1%). Levels of estradiol (E2), testosterone (T), luteinzing hormone (LH), dehydroiso-androsterone (DHEA-S) and prolactin (PRL) were higher, while the level of sexual hormone binding protein (SHBG) was lower in PCOS patients than those in control, follicular stimulating hormone (FSH) level in patients of Shen-deficiency syndrome and phlegm-dampness syndrome was high than that in control (P < 0.05 and P < 0.01). However, no significant differences were found in comparing the various sexual endocrinal indices between patients with different syndrome types (P > 0.05). Besides, the level of PRL was positively correlated with LH and E2 levels in patients. CONCLUSION: Chinese medicine syndromes presented in patients with PCOS are mostly intermingling, Shen-deficiency and Gan-stagnancy are the basic syndromes, and there is some correlation between syndrome type and sexual hormone levels.

[Effect of cyclic GMP-AMP synthase on EMT in breast cancer cells].

OBJECTIVE: To investigate the effects of cyclic GMP-AMP synthase (cGAS) on the proliferation, migration and epithelial to mesenchymal transition (EMT) of breast cancer cells. METHODS: The cGAS lentiviral vector and control fluorescence vector were transfected into breast cancer MCF7 cells and were divided into negative group (NC) and MCF7-cGAS group. The effect of cGAS on proliferation in the MCF7 cells was detected by MTT. The effect of cGAS on cell migration was detected by Transwell assay. The expressions of EMT related proteins were analyzed by Western blot. RESULTS: After over-expressed with cGAS, the proliferation and migration of MCF7 cells were increased (P＜0.05). The expression level of the epithelial markers E-cadherin was decreased, while the expression level of the mesenchymal markers N-cadherin was increased(P＜0.05). CONCLUSION: The over-expression of cGAS increased the proliferation and migration of breast cancer cells and induced EMT in breast cancer cells.

SNCA-Rep1 polymorphism correlates with susceptibility and iron deficiency in restless legs syndrome.

BACKGROUND: Brain iron disequilibrium and dopaminergic dysfunction are key pathophysiological features of Restless Legs Syndrome (RLS). Rep1 polymorphism in the promotor region of SNCA is associated with risk of Parkinson's disease, however its association with RLS and iron status is unclear. OBJECTIVE: To investigate SNCA-Rep1 polymorphism in RLS and its phenotypes. METHODS: We recruited RLS patients as well as age and gender matched healthy controls. Demographic information and clinical features of RLS were recorded. Laboratory examinations were performed to exclude possible secondary causes. RESULTS: 215 RLS patients and 369 healthy controls were included. We found that the Rep1 allele 0 homozygosity significantly decreased RLS risk (OR: 0.345; P < 0.0001, and remained significant after the Bonferroni correction). Phenotypic analysis demonstrated that longer Rep1 alleles were associated with increased susceptibility to iron deficiency (53.0% vs 36.1%, P = 0.017), however had no phenotypic significant effects on age, gender, onset age, duration, RLS family history, severity, laterality, extra body involvement and seasonal fluctuation. Multivariate logistic regression analyses confirmed long Rep1 allele was associated with higher risk of iron deficiency in RLS after adjusting for potential confounding factors. In detail, Rep1 allele 2 homozygosity was prone to a higher risk of peripheral iron deficiency in RLS (OR: 4.550, P = 0.006, remained significant after the Bonferroni correction). CONCLUSION: The SNCA-Rep1 variability modified RLS risk and influenced peripheral iron deficiency in this group of Chinese RLS patients. Rep1 allele 0 homozygosity decreased the risk of RLS, while homozygous allele 2 increased the risk of nonanemic iron deficiency in RLS.

Operative timing of liver transplantation for patients with severe hepatitis.

BACKGROUND: Fulminant hepatic failure manifests a rapid onset, serious complications, and a high mortality, but still there is a possibility of recovery. Once the patient is able to pass a crisis, the liver is able to regenerate completely and regain its normal function. Therefore it is of vital importance to determine the eligible timing for transplantation. Premature surgery might result in a loss of the chance of internal medical treatment and misuse of liver resources, whereas delayed surgery might increase the difficulty of treatment in the preoperative period and the possibility of complications and medical expense, which eventually result in decreased rate of success and survival. This problem remains worldwide how to choose the optional timing of operation. METHODS: Thirty-six patients with severe hepatitis were treated by orthotopic liver transplantation. The distribution of MELD scores in these patients was: 10-19 in 8 patients, 20-29 in 10, 30-39 in 11, and 40 in 7. They were divided into two groups: MELD score <30 and MELD score >or=30. Parameters (1-year survival rate, complications, preoperative use of artificial liver, operative time, volume of bleeding and blood transfusion, and average hospital costs) were examined as prognostic factors after liver transplantation. RESULTS: The 1-year survival rate of the MELD score <30 group was higher than that of the >or=30 group (77.8% and 33.3%, P=0.007), and the rate of complications in the <30 group was lower (P=0.012). There were no differences in the timing of artificial liver treatment, operative time, operative hemorrhage, and transfusion between the two groups (P=0.742). But the average daily hospital cost in the MELD score >or=30 group was higher (P=0.008). CONCLUSION: This study shows that when the MELD score is <30 it may be the optimal time to perform liver transplantation for patients with severe hepatitis.

Paroxysmal movement disorders: Recent advances and proposal of a classification system.

The increasing recognition of the phenotypic and genotypic heterogeneity that exists amongst the paroxysmal movement disorders (PMDs) is challenging the way these disorders have been traditionally classified. The present review aims to summarize how recent genetic advances have influenced our understanding of the nosology, pathophysiology and treatment strategies of paroxysmal movement disorders. We propose classifying PMDs using a system that would combine both phenotype and genotype information to allow these disorders to be better categorized and studied. In the era of next generation sequencing, the use of a standardized algorithm and employment of selective genetic screening will lead to greater diagnostic certainty and targeted therapeutics for the patients.