A systematic meta-analysis of immune signatures in patients with COVID-19.

Currently severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission has been on the rise worldwide. Predicting outcome in COVID-19 remains challenging, and the search for more robust predictors continues. We made a systematic meta-analysis on the current literature from 1 January 2020 to 15 August 2020 that independently evaluated 32 circulatory immunological signatures that were compared between patients with different disease severity was made. Their roles as predictors of disease severity were determined as well. A total of 149 distinct studies that evaluated ten cytokines, four antibodies, four T cells, B cells, NK cells, neutrophils, monocytes, eosinophils and basophils were included. Compared with the non-severe patients of COVID-19, serum levels of Interleukins (IL)-2, IL-2R, IL-4, IL-6, IL-8, IL-10 and tumor necrosis factor α were significantly up-regulated in severe patients, with the largest inter-group differences observed for IL-6 and IL-10. In contrast, IL-5, IL-1ß and Interferon (IFN)-γ did not show significant inter-group difference. Four mediators of T cells count, including CD3+ T, CD4+ T, CD8+ T, CD4+ CD25+ CD127- Treg, together with CD19+ B cells count and CD16+ CD56+ NK cells were all consistently and significantly depressed in severe group than in non-severe group. SARS-CoV-2 specific IgA and IgG antibodies were significantly higher in severe group than in non-severe group, while IgM antibody in the severe patients was slightly lower than those in the non-severe patients, and IgE antibody showed no significant inter-group differences. The combination of cytokines, especially IL-6 and IL-10, and T cell related immune signatures can be used as robust biomarkers to predict disease severity following SARS-CoV-2 infection.

Dexamethasone suppresses the growth of human non-small cell lung cancer via inducing estrogen sulfotransferase and inactivating estrogen.

AIM: Dexamethasone (DEX) is a widely used synthetic glucocorticoid, which has shown anti-cancer efficacy and anti-estrogenic activity. In this study we explored the possibility that DEX might be used as an endocrine therapeutic agent to treat human non-small cell lung cancer (NSCLC). METHODS: The viability and proliferation of human NSCLC cell lines A549 and H1299 were assessed in vitro. Anti-tumor action was also evaluated in A549 xenograft nude mice treated with DEX (2 or 4 mg·kg(-1)·d(-1), ig) or the positive control tamoxifen (50 mg·kg(-1)·d(-1), ig) for 32 d. The expression of estrogen sulfotransferase (EST) in tumor cells and tissues was examined. The intratumoral estrogen levels and uterine estrogen responses were measured. RESULTS: DEX displayed mild cytotoxicity to the NSCLC cells (IC50 >500 µmol/L) compared to tamoxifen (IC50 <50 µmol/L), but it was able to inhibit the cell proliferation at low micromolar ranges. Furthermore, DEX (0.1-10 µmol/L) dose-dependently up-regulated EST expression in the cells, and inhibited the cell migration in vitro. Triclosan, a sulfation inhibitor, was able to diminish DEX-caused inhibition on the cell viability. In A549 xenograft nude mice, DEX or tamoxifen administration remarkably suppressed the tumor growth. Moreover, DEX administration dose-dependently increased EST expression in tumor tissues, and reduced intratumoral estrogen levels as well as the volumes and weights of uterine. CONCLUSION: DEX suppresses the growth of A549 xenograft tumors via inducing EST and decreasing estradiol levels in tumor tissues, suggesting that DEX may be used as anti-estrogenic agent for the treatment of NSCLC.

Preclinical PK/PD model for combined administration of erlotinib and sunitinib in the treatment of A549 human NSCLC xenograft mice.

AIM: Combined therapy of EGFR TKI and VEGFR TKI may produce a greater therapeutic benefit and overcome EGFR TKI-induced resistance. However, a previous study shows that a combination of EGFR TKI erlotinib (ER) with VEGFR TKI sunitinib (SU) did not improve the overall survival in patients with non-small-cell lung cancer (NSCLC). In this study we examined the anticancer effect of ER, SU and their combination in the treatment of A549 human NSCLC xenograft mice, and conducted PK/PD modeling and simulations to optimize the dose regimen. METHODS: ER (20, 50 mg·kg(-1)·d(-1)) or SU (5, 10, 20 mg·kg(-1)·d(-1)) alone, or their combination were administered to BALB/c nude mice bearing A549 tumors for 22 days. The tumor size and body weight were recorded daily. The experimental data were used to develop PK/PD models describing the quantitative relationship between the plasma concentrations and tumor suppression in different dose regimens. The models were further evaluated and validated, and used to predict the efficacy of different combination regimens and to select the optimal regimen. RESULTS: The in vivo anticancer efficacy of the combination groups was much stronger than that of either drug administered alone. A PK/PD model was developed with a combination index (φ) of 4.4, revealing a strong synergistic effect between ER and SU. The model simulation predicted the tumor growth in different dosage regimens, and showed that the dose of SU played a decisive role in the combination treatment, and suggested that a lower dose of ER (≤5 mg·kg(-1)·d(-1)) and adjusting the dose of SU might yield a better dosage regimen for clinical research. CONCLUSION: The experimental data and modeling confirm synergistic anticancer effect of ER and SU in the treatment of A549 xenograft mice. The optimal dosage regimen determined by the PK/PD modeling and simulation can be used in future preclinical study and provide a reference for clinical application.

How Work Organization Affects the Prevalence of WMSDs: A Case-control Study.

OBJECTIVE: In this study, we aimed at exploring the association between work-related musculoskeletal disorders (WMSDs) and work organization based on a case-control study. METHODS: A total of 1938 workers who claimed to suffer from WMSDs were selected from Beijing, Henan, Hubei, and the Guangdong province. The control group consisted of 2009 workers employed in similar industries without severe disease or musculoskeletal discomforts. We used a modified version of the questionnaire developed by the NMQ and the DMQ to investigate individual and work-related factors. RESULTS: A total of 13 variables (P<0.1) were selected by the chi-square test and finally, 7 variables entered into the equation, with 6 variables reaching statistical significance (P<0.05). The odds ratios (OR) of 'work changing with season' and 'sufficient rest time' did not reach 1 (0.749 and 0.441, respectively). In addition, 'sufficient rest time' seemed to be the stronger protective factor according to its higher standardized coefficient. And 'repetitive work every minute', 'constantly repetitive work' (every day), 'shortage of site personnel', and 'often switching shifts with others' seemed to be the risk factors. CONCLUSION: Work organization may have comprehensive effects on the occurrence of WMSDs. This pattern of associations suggests that further investigation into the mechanism of how work organization affects the prevalence of WMSDs is required.

Design, synthesis, and biological evaluation of novel trifluoromethyl indoles as potent HIV-1 NNRTIs with an improved drug resistance profile.

A novel series of trifluoromethyl indole derivatives have been designed, synthesized and evaluated for anti-HIV-1 activities in MT-2 cells. The hydrophobic constant, acute toxicity, carcinogenicity and mutagenicity were predicted. Trifluoromethyl indoles 10i and 10k showed extremely promising activities against WT HIV-1 with IC50 values at the low nanomolar level, similar to efavirenz, better than nevirapine, and also possessed higher potency towards the drug-resistant mutant strain Y181C than nevirapine. Preliminary SAR and docking studies of detailed binding mode provided some insights for discovery of more potent NNRTIs.

Metal-enhanced fluorescence of nano-core-shell structure used for sensitive detection of prion protein with a dual-aptamer strategy.

Metal-enhanced fluorescence (MEF) as a newly recognized technology is widespread throughout biological research. The use of fluorophore-metal interactions is recognized to be able to alleviate some of fluorophore photophysical constraints, favorably increase both the fluorophore emission intensity and photostability. In this contribution, we developed a novel metal-enhanced fluorescence (MEF) and dual-aptamer-based strategy to achieve the prion detection in solution and intracellular protein imaging simultaneously, which shows high promise for nanostructure-based biosensing. In the presence of prion protein, core-shell Ag@SiO2, which are functionalized covalently by single stranded aptamer (Apt1) of prions and Cyanine 3 (Cy3) decorated the other aptamer (Apt2) were coupled together by the specific interaction between prions and the anti-prion aptamers in solution. By adjusting shell thickness of the pariticles, a dual-aptamer strategy combined MEF can be realized by the excitation and/or emission rates of Cy3. It was found that the enhanced fluorescence intensities followed a linear relationship in the range of 0.05-0.30 nM, which is successfully applied to the detection of PrP in mice brain homogenates.

Diverse reactivity in microwave-promoted catalyst-free coupling of substituted anilines with ethyl trifluoropyruvate and biological evaluation.

Diverse reactivity by coupling of substituted anilines with ethyl trifluoropyruvate was developed under microwave irradiation without catalysts to generate 3-trifluoromethyl-3-hydroxy oxindoles, aromatic hydroxy trifluoromethyl esters, and 1,2-dicarbonyl compounds in a fast and efficient manner. The plausible mechanism for obtaining different products was proposed. Furthermore, the anti-HIV activity of aromatic hydroxy trifluoromethyl esters was first reported. The best inhibitory activity against wild-type HIV-1 IIIB was exemplified by trifluoromethyloxindole 3q with an IC50 = 5.8 µM, which also displayed potential activity against Y181C mutant virus with an IC50 = 7.5 µM. More significantly, the activities of oxindoles 3q and 3r to inhibit K103N/Y181C double mutant HIV-1 reverse transcriptase (RT) are probably similar to that of the second-generation nonnucleoside inhibitor HBY 097 by docking calculation.

[A cross-sectional study on the prevalence of HIV drug resistance in patients receiving antiretroviral treatment in Shenqiu county, Henan province].

OBJECTIVE: To understand the prevalence of drug resistance in AIDS patients who had been receiving HAART in a long run, in Shenqiu county, Henan province. METHODS: This cross-sectional study included 120 HIV infected patients who began receiving ART (antiretroviral therapy) in 2003. Viral loads and CD(4)(+) T cells counts were measured, and In-house drug resistance test was performed in VL > 1000 copies/ml patients. RESULTS: 114 cases out of 120 patients had complete viral load data. Among them, 33 cases having viral loads less than 50 copies/ml, and the remaining viral loads showed an average of lg (4.09 ± 1.10) copies/ml. The average of CD(4)(+) T cell counts was (377 ± 218) cells/ml, with 64 (53.3%) cases showing their CD(4)(+) T cell counts higher than 350 cells/ml. In 67 patients, 58 of them showed genotypic resistance, and 40 cases showed reverse transcriptase inhibitors (RTIs) resistance. The ratios of nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs) resistance were 53.4% (31/58) and 67.2% (39/58), respectively. There were no differences of drug resistance ratio in the three treatment programs. The highest drug resistance rates in NRTIs and NNRTIs were zidovudine, lamivudine, nevirapine. However, protease inhibitors (PIs) resistance variants were not found. CONCLUSION: The prevalence of drug-resistant strains seemed to be high in Shenqiu country, Henan province. Long-term follow-up monitoring strategy should be developed to optimize the timely treatment programs.

[Distribution of HIV-1 subtypes in Guangxi Zhuang Autonomous Region, 2008-2009].

OBJECTIVE: To understand the distribution of HIV-1 subtypes in Guangxi Zhuang Autonomous Region. METHODS: 294 participants who were infected by HIV-1 in 2008 - 2009 and residing in 13 cities in Guangxi were enrolled into this study. Epidemiological information showed that heterosexual transmission was the main transmitting route. 1584 bp full length gag gene, 3147 bp full length pol gene and 558 bp env (C2V3) fragments were amplified by using RT-PCR and then directly sequenced. NCBI genotyping tool and Mega 5.03 software were used to determine the HIV subtypes. Simplot and Recombinant HIV-1 Drawing Tool were used for the analysis of recombination. RESULTS: A total of 739 sequences, including 270 gag, 246 pol and 223 env (C2V3), were successfully obtained from 294 plasma specimens. Genotypes of HIV from 272 participants were determined. CRF01_AE was found the dominant (77.6%), followed by CRF08_BC (10.7%) and CRF07_BC (7.4%). 7 unique recombinant forms, 4 subtype B (B') and 1 subtype G were also identified. No significant difference on the distribution of subtypes among different regions and ethnics was found. Among the 7 unknown recombinant forms, 6 strains were mosaic B and/or C fragments with CRF01_AE genome as the backbone, while one strain originated from CRF07_BC and CRF08_BC. CONCLUSION: Currently, CRF01_AE was found the major subtype of HIV epidemic in Guangxi. New recombinant forms with CRF01_AE as backbones had been emerging, which called for serious attention.

A click-activated fluorescent probe for selective detection of hydrazoic acid and its application in biological imaging.

Here we present a new approach to aqueous hydrazoic acid detection through the synthesis and evaluation of an alkyne-based fluorescent probe, which could be applied in the monitoring of hydrazoic acid in both living Hela cells and larval zebrafish. This probe can also serve as an early warning automaton which would alert when both azide and protons exist over the threshold value.

Risk factors of low back pain among the Chinese occupational population: a case-control study.

OBJECTIVE: To explore the risk factors of low back pain among the Chinese occupational population in several major industries. METHODS: A total of 7200 subjects (3600 cases and 3600 controls) were randomly sampled from a cross-sectional study, and they were investigated for individual and occupational factors of low back pain. The potential risk factors were first selected by using chi-square tests. Secondly, collinearity diagnosis proceeded by using the Kendall's rank correlation. Finally, binary logistic regression model was used for multi-factor analysis. RESULTS: Collinearity diagnosis showed that there was a severe collinearity problem among the potential risk factors of low back pain. Logistic regression model included 20 variables with statistical significance. Bending neck forward or holding neck in a forward posture for long periods (OR=1.408) was the most important risk factor inducing low back pain in this study, followed by bending heavily with the trunk (OR=1.402), carrying out identical work almost for the whole day (OR=1.340). Additionally, sufficient normal break was a protective factor of low back pain. CONCLUSION: Low back pain among the Chinese occupational population was associated with body height, occupation, work organization, physical work, working posture, and others. All these risk factors could be regarded as the indicators of low back pain, and some relevant preventive measures should be taken to reduce low back pain risk.

Effect of VEGF-targeted antisense gene therapy on retinoblastoma cell line SO-RB50 in vitro and in vivo.

AIM: To evaluate the possibility of generation 4 polyamidoamine (G4PAMAM) dendrimers acting as the delivery system of vascular endothelial growth factor (VEGF) antisense oligodeoxynucleotides (VEGFASODN), and to investigate the anti-tumor effect of G4PAMAM/VEGFASODN complex on the cultured cells and the mouse tumor xenograft model. METHODS: The transfection efficiency was assessed by Flow cytometry (FCM). Thiazolyl tetrazolium (MTT) assay was performed to determine the relative growth rate (RGR) of the cells after transfection. Then a mouse tumor xenograft model of human retinoblastoma was established. Different interventions were given to the mice by intratumoral injection and the tumor growth was monitored. The expression of VEGF mRNA was detected by reverse transcription PCR (RT-PCR), the expression of VEGF protein was determined by western blot analysis, and the microvessel density (MVD) was measured by immunohistochemistry (IHC) staining. RESULTS: G4PAMAM/VEGFASODN exhibited a high transfection rate in vitro, and the transfection rates of different doses of G4PAMAM/VEGFASODN groups increased with higher doses. This effect was accompanied by a dose-depended reduction in cell viability. The tumor growth in the tumor-bearing athymic mice was significantly inhibited in the G4PAMAM/VEGFASODN group. The expressions of VEGF mRNA and protein were obviously inhibited in the G4PAMAM/VEGFASODN group (P<0.05), and the MVD of the G4PAMAM/VEGFASODN group was lower than that of the other groups (P<0.05). CONCLUSION: VEGFASODN can be delivered into the cultured and transplanted retinoblastoma cells efficiently by G4PAMAM, suppress the expressions of VEGF mRNA and protein, and reduce the MVD of tumor tissues. The G4PAMAM/VEGFASODN complex has antitumor properties in vitro and in vivo.

[HIV-1 subtype and the distribution in Yunnan province].

OBJECTIVE: To understanding the genetic subtype and its population and regional distribution of HIV-1 strains circulating in Yunnan province. METHODS: 788 plasma specimens collected in 2008-2009 from 15 distracts of Yunnan, were enrolled. Viral RNA were extracted and subjected to RT-PCR. 1584 bp full length gag gene, 3147 bp full length pol gene and 558 bp env (C2V3) fragment were amplified and directly sequenced. Full length gag and pol genes were connected together as a complete genetic region (location on HXB2: 790 - 5096) for genotyping. RESULTS: Of the 788 plasma specimens, a total number of 1728 genomic sequences including 599 gag, 564 pol and 525 env (C2V3) were successfully amplified and sequenced, with genotype of 617 samples identified. The subtypes of HIV-1 strains circulated in Yunnan were with the order of constituent ratio CRF08_BC (50.2%), CRF01_AE (25.0%), unknown recombinant forms (10.2%), CRF07_BC (9.2%), subtype C (2.9%) and subtype B (B') (2.4%). The distributions of subtypes showed significant regional differences and could be roughly divided into two forms:the CRF08_BC predominant areas represented by Lincang and Kunming, and the areas with CRF08_BC together with CRF01_AE coexistence, represented by Dehong and Xishuangbanna. The unknown recombinant forms accounted for more HIV infection in ethnic minorities (17.0%) than in ethnic Han (6.7%, P < 0.01). The distribution of subtypes varied significantly in the two primary routes of transmission for those infected through heterosexual contact. CRF08_BC and CRF01_AE were the dominant subtypes, accounting for 52.7% and 29.1% respectively. However, in IDUs, CRF08_BC strains accounted for half of the infection, while only 4.5% of the infections were caused by CRF01_AE, CRF07_BC while the unique recombinant forms were responsible for 15.5% infections. Of the 63 unknown recombinant forms, most (74.6%) were B (B') recombinant with C, while 25% were mosaic B and/or C fragments on the bases of CRF01_AE genome. CONCLUSION: The subtypes of HIV-1 strains circulated in Yunnan were complicated under the significant differences of regions, ethnics or routes of transmission.

[The assessment of reliability and validity of musculoskeletal questionnaire].

OBJECTIVE: To evaluate the reliability and validity of musculoskeletal questionnaire. METHODS: A self-administered modified musculoskeletal questionnaire was used to investigate 12 098 workers from eight occupations, i.e. coal mining, petroleum, metallurgical, mechanical manufacturing, chemical, garment and railroad transportation industries and education. The Cronbach's α coefficient, analysis of covariance and multiple logistic regression were used to assess the reliability and validity of musculoskeletal questionnaire. RESULTS: The consistent test between total items of Musculoskeletal Questionnaire and each factor showed that the range of Cronbach's α was 0.52 ∼ 0.92, except from vibration factor, other Cronbach's α was more than 0.7. All 55 items of Musculoskeletal Questionnaire were subjected to factor analysis, and ten latent factors were identified, which explained 55.17% of the total variance. The potentially hazardous working conditions could be categorized into seven dimensions (force, dynamic load, static load, repetitive load, climate factors, vibration exposure and environmental ergonomic factor), which consisted with the theory model. The results of covariance analysis indicated that there were significant difference among 7 dimension indices in different jobs (P < 0.01). CONCLUSION: The modified Musculoskeletal Questionnaire is a valid and reliable tool for measuring musculoskeletal workload.

Similar neutralizing activity in the HIV-1 infected long term non-progressors(LTNPs) and typical progressors(TPs).

Neutralizing antibodies are considered to be an important protective parameter used in HIV-1 vaccine evaluation. However, the exact role that neutralizing antibodies plays in controlling the disease progression of HIV-1 infected peoples is still undetermined. In this paper, we compared the protective function of the neutralizing antibody response in the plasma from LTNP and TP against clade B and clade C pseudoviruses. No difference in the neutralizing activities between the plasma from LTNP and TP was found, which was consistent with the most recent reports. In addition, no correlations between the titer or breadth and CD(4+) or viral load in HIV-1 infected individuals were found. The protective roles played by neutralizing antibodies in controlling disease progression of HIV-1 infected people need to be considered in a new viewpoint.

[Prevalence status and mutation pattern of H221Y in subtype B' of HIV-1].

OBJECTIVE: To explore the prevalence and mutation pattern of H221Y at reverse transcriptase (RT) among the subtype B' of human immunodeficiency virus1 (HIV-1) in antiviral therapy-failure patients. METHODS: A total of 1363 sequences, comprising of 1205 therapy-failure individuals and 158 therapy-naive individuals, were submitted to the Stanford HIV drug resistance database (SHDB) to analyze the frequency and mutation pattern of H221Y. RESULTS: The prevalence of mutation H221Y in the therapy-failure population was significantly higher than that of the therapy-naive (6.59% vs 0.60%) (χ(2) = 6.59, P = 0.027). The emergence of H221Y usually accompanied the position mutations of T215, M184, K103 and Y181 of RT, and the pattern of TAMs/H221Y/Y181C/I was common. Frequency of H221Y in the regimen of AZT/ddI/NVP was more popular than the other 4 regimens (14.6% vs 3.5%, 4.9%, 2.3%, 2.6%, all P < 0.01). CONCLUSION: With a unique mutation pattern, H221Y has a low prevalence in the individuals of first-line therapy-failure patients.

CD3Z genetic polymorphism in immune response to hepatitis B vaccination in two independent Chinese populations.

Vaccination against hepatitis B virus is an effective and routine practice that can prevent infection. However, vaccine-induced immunity to hepatitis B varies among individuals. CD4(+) T helper cells, which play an important role in both cellular and humoral immunity, are involved in the immune response elicited by vaccination. Polymorphisms in the genes involved in stimulating the activation and proliferation of CD4(+) T helper cells may influence the immune response to hepatitis B vaccination. In the first stage of the present study, a total of 111 single nucleotide polymorphisms (SNPs) in 17 genes were analyzed, using the iPLEX MassARRAY system, among 214 high responders and 107 low responders to hepatitis B vaccination. Three SNPs (rs12133337 and rs10918706 in CD3Z, rs10912564 in OX40L) were associated significantly with the immune response to hepatitis B vaccination (P = 0.008, 0.041, and 0.019, respectively). The three SNPs were analyzed further with the TaqMan-MGB or TaqMan-BHQ probe-based real-time polymerase chain reaction in another independent population, which included 1090 high responders and 636 low responders. The minor allele 'C' of rs12133337 continued to show an association with a lower response to hepatitis B vaccination (P = 0.033, odds radio = 1.28, 95% confidence interval = 1.01-1.61). Furthermore, in the stratified analysis for both the first and second populations, the association of the minor allele 'C' of rs12133337 with a lower response to hepatitis B vaccination was more prominent after individuals who were overweight or obese (body mass index ≥25 kg/m(2)) were excluded (1(st) stage: P = 0.003, 2(nd) stage: P = 0.002, P-combined = 9.47e-5). These findings suggest that the rs12133337 polymorphism in the CD3Z gene might affect the immune response to hepatitis B vaccination, and that a lower BMI might increase the contribution of the polymorphism to immunity to hepatitis B vaccination.

Fluorescence turn-on detection of hypochlorous acid via HOCl-promoted dihydrofluorescein-ether oxidation and its application in vivo.

We report three new water-soluble dihydrofluorescein-ether probes FCN1, FCN2 and FCN3 for the detection of hypochlorous acid (HOCl), which were designed on the basis of a specific HOCl-promoted oxidation reaction. This work also provided a useful method to monitor the accumulated HOCl in specific organelles using a zebrafish model.

Mutations in PRRT2 result in paroxysmal dyskinesias with marked variability in clinical expression.

BACKGROUND: Paroxysmal dyskinesias (PDs), a clinically and genetically heterogeneous group of episodic movement disorders, include kinesigenic PD (PKD), exercise-induced PD (PED) and non-kinesigenic PD (PNKD). These disorders are all transmitted as autosomal dominant traits with incomplete penetrance. Several PD-related genetic disorders, including PKD and familial infantile convulsions with paroxysmal choreoathetosis (ICCA), mapped to the same region on chromosome 16. Independent genetic studies have recently revealed that PKD can be caused by loss-of-function mutations in the proline-rich transmembrane protein 2 gene (PRRT2). We tested the hypothesis that other forms of PDs are also due to PRRT2 mutations. METHODS/RESULTS: The whole genomic region of PRRT2 was sequenced in six Han Chinese families and 15 sporadic cases of PD-related phenotypes. The previously reported mutation, c.649dupC (p.R217Pfs*7), was found in two families with PKD, one family with ICCA, one family with PNKD-like phenotype, and two sporadic cases with PED. In an additional ICCA family, a novel frameshift mutation, c.904dupG (p.D302Gfs*38), was identified. A missense mutation, c.913G→A (p.G305R), and a synonymous substitution, c.1011C→T (p.G337G), were also detected in two sporadic PKD cases. CONCLUSION: This study shows that PKD, ICCA and some other PD-related phenotypes are part of the same phenotypic spectrum, caused by mutations in PRRT2. This underscores the complexity of the phenotypic consequences of PRRT2 mutations.