[Correlation study between changes in intestinal microflora structure and immune indexes in newly treated patients with pulmonary tuberculosis].

To explore the correlation between the changes of the intestinal flora of newly treated pulmonary tuberculosis patients and the immune indicators of the body, and to provide a reference for the prevention and treatment of pulmonary tuberculosis. A single-center and case-control study was adopted. From October 2020 to April 2021, 43 patients with newly diagnosed tuberculosis in the Department of Tuberculosis, Affiliated Changsha Central Hospital,University of South China were selected as the control group. 43 cases of newly treated pulmonary tuberculosis (PTB), 43 healthy control (HC) during the same period, collected fresh feces and whole blood of subjects, and used Illumina Hiseq high-throughput sequencing technology to analyze 16S of all microorganisms in feces The V4 region of rRNA was amplified and sequenced, and the structure of the intestinal flora was analyzed by QIIME software. Use flow cytometry to determine the subject's immune indicators (CD3+, CD4+, CD8+, CD4+CD25+CD127-Treg, CD14+CD16+, CD14+CD16-), and analyze the changes in intestinal flora and immune function in newly treated pulmonary tuberculosis patients Inherent connection. The χ² test, t test, and Wilcox rank sum test were used to analyze the differences in age, gender, α diversity, and relative abundance of the two groups of people. Compared with the HC group, the alpha diversity of the intestinal flora in the PTB group decreased (shannon index: t=3.906, P=0.000 2; simpson index: Z=553, P=0.004 7; chao1 index: t=5.395, P=0.000 0). ß diversity analysis showed that there were significant differences in the structure of the intestinal flora between the two groups (P=0.000). Species difference analysis showed that at the phylum level, the relative abundance of Firmicutes in the PTB group was significantly lower than that in the HC group (Z=486.0, P=0.000 5). At the genus level, there are 15 different bacterial genera between the two groups. In the PTB group, bifidobacterium, enterococcus, lactobacillus, anaerostipes, the relative abundance of the above 5 genera of veillonella is higher than that of the HC group (P<0.05); Butyricimonas, clostridium, and broutella (blautia), coprococcus, dorea, lachnospira, roseburia, faecalibacterium, ruminococcus, the relative abundance of 10 bacterial genera including dialister was lower than that of the HC group (P<0.05). Comparison of immune indexes between groups showed that CD14+CD16+monocytes (%) in the PTB group were higher than those in the HC group (t=2.456, P=0.001 6<0.05), while CD14+CD16-monocytes (%) were lower than HC (t=-4.368, P=0.000<0.05), while the differences in CD3+, CD4+, CD8+, CD4+/CD8+and Treg (CD4+CD25+CD127-) were not statistically significant (P>0.05). Spearman correlation analysis showed that Firmicutes in the PTB group was negatively correlated with CD4+/CD8+, CD14+CD16+(r=-0.218, P=0.048; r=-0.245, P=0.025), and positively correlated with CD14+CD16-Correlation (r=0.250, P=0.022); At the genus level, Faecalis is positively correlated with CD4+/CD8+and CD4+(r=0.250, P=0.023; r=0.258, P=0.019); Rosella and CD3+, CD8+and CD14+CD16-are positively correlated (r=0.27, P=0.024; r=0.219, P=0.046; r=0.027, P=0.039), and negatively correlated with CD14+CD16+(r=-0.280, P= 0.01). Changes in the structure of the intestinal flora of newly treated pulmonary tuberculosis patients may be one of the influencing factors of the immune function of the body. Targeted optimization of the structure of the intestinal flora and improvement of the body's immunity may be used as an effective auxiliary treatment for pulmonary tuberculosis.

[Changes of Rheb gene and protein expression in preeclampsia-like mouse model treated with pravastatin].

Objective: To explore whether pravastatin (Pra) inhibits mammalian target of rapamycin (mTOR) signal pathway by regulating Ras homolog enriched in brain (Rheb) protein through the comparison of gene and protein expression changes of Rheb in liver and placenta in preeclampsia (PE)-like mouse model treated with Pra. Methods: C57BL/6J pregnant mice were randomly divided into two groups. The PE group was established by injecting N-nitro-L-arginine methyl ester (L-NAME) daily at gestational 7-18 days, saline was injected as contol group (Con); then giving mice Pra (PE+Pra, Con+Pra group, n=8) or normal saline (PE+N, Con+N group, n=8) every day from the 8th gestational day of pregnancy. The maternal liver and placenta tissues were collected on the 18th day of pregnancy. Western blot, real-time quantitative PCR and immunohistochemistry were used to compare the levels of Rheb protein and mRNA expression in the liver and placenta. Results: (1)The results of western blot: there were no significant differences in Rheb protein expression between PE+N group (liver: 0.706±0.123; placenta: 0.866±0.128) and Con+N group (liver: 0.732±0.123; placenta: 0.909±0.097) , and the differences between PE+Pra group (liver: 0.669±0.134; placenta: 0.940±0.221) and PE+N group were not significant either in liver or in placenta (all P>0.05). (2) The results of real-time quantitative PCR: when PE+N group (liver: 1.026±0.480; placenta: 1.102±0.361) compared with Con+N group (liver: 1.058±0.389; placenta: 1.067±0.400) , PE+Pra group (liver: 0.735±0.356; placenta: 0.822±0.304) compared with PE+N group, there were no significant differences either in liver or in placenta (all P>0.05). (3) The results of immunohistochemistry: Rheb protein expression did not change significantly in maternal liver and placenta, there were no significant differences in protein expression levels between PE+N group and Con+N group, and between PE+Pra group and PE+N group (all P>0.05). Conclusion: The inhibition of Pra on mTOR signaling pathway in some PE-like model may be independent of the expression of Rheb gene and protein.

[Regulation of pravastatin on long-chain fatty acid oxidative enzyme in pre-eclampsia-like mouse model].

Objective: To investigate the modulation of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) expression by pravastatin in pre-eclampsia-like mouse model. Methods: C57BL/6J mice were randomly injected with N-nitro-L-arginine methyl ester (L-NAME) as pre-eclampsia-like model group (PE) or saline as normal pregnancy control group (Con) respectively, from gestational the 7th to 18th day. For each group, pravastatin (PE+Pra, Con+Pra group) or saline (PE+N, Con+N Group) was given from the 8th to 18th day of gestation, respectively. Liver and placenta of pregnant mice were collected on gestational day 18. The LCHAD protein expression and mRNA levels of liver and placenta were detected through western blot, immunohistochemistry and real-time quantitative PCR. Results: (1) The average arterial pressure of pregnant mice increased gradually from the 8th to 18th day in PE+N group, but decreased in PE+Pra group from gestational 10th day, 24 hour urinary protein levels in PE+N group [(1 494 ± 201) µg] were significantly higher than that in Con+N group [(935±128) µg, P<0.01], and also higher than that in PE+Pra group [(981±116) µg, P<0.01].(2) The results of western blot: the expression of LCHAD was significantly lower in PE+N group (liver: 0.64±0.11, placenta: 0.48±0.06) than that in Con+N group (liver: 1.06±0.10, placenta: 0.60±0.10), and lower than that in PE+Pra group (liver: 0.99±0.04, placenta: 0.60±0.08; all P<0.01).(3)The results of real-time quantitative PCR: the levels of LCHAD mRNA in liver and placenta in PE+N group (liver: 0.621±0.128, placenta: 0.646±0.129) were significantly decreased compared with Con+N group (liver: 1.007±0.130, placenta: 1.004±0.103; all P<0.01), but there was no significant difference between PE+Pra group (liver: 0.693±0.678, placenta: 0.662±0.183; P>0.05). (4) LCHAD protein was expressed widely and evenly in liver. The expression in placental cytotrophoblast and syncytial trophoblast cells located in outer layer of villous in labyrinth layer was the most. The expression of LCHAD was significantly lower in PE+N group (liver: 0.062±0.016, placenta: 0.147±0.018) than that in Con+N group (liver: 0.126±0.013, placenta: 0.183±0.024), and lower than that in PE+Pra group (liver: 0.111±0.017, placenta: 0.174±0.027; all P<0.05). Conclusion: Pravastatin could upregulate the LCHAD protein expression of liver and placenta in the pre-eclampsia-like mouse, which may be a mechanism to improve the clinical manifestations of pre-eclampsia.

Echinoside A, a new marine-derived anticancer saponin, targets topoisomerase2alpha by unique interference with its DNA binding and catalytic cycle.

BACKGROUND: Echinoside A was isolated from sea cucumber. This study demonstrates its anticancer effects and its mechanisms of action. MATERIALS AND METHODS: Anticancer effects of echinoside A were evaluated in vitro and in vivo. TUNEL and DNA fragmentation assays were applied to examine its ability to induce apoptosis. A series of biochemical assays were applied to investigate the inhibition of echinoside A on topoisomerase2alpha (Top2alpha). Molecular docking analyses were used to demonstrate the direct interaction between echinoside A and Top2alpha. RESULTS: Echinoside A inhibited the growth of tumors in mouse models and human prostate carcinoma xenografts in nude mouse models. Echinoside A shows the unique characteristics of inhibiting the noncovalent binding of Top2alpha to DNA by competing with DNA for the DNA-binding domain of the enzyme and of interfering predominantly with the Top2alpha-mediated prestrand passage cleavage/religation equilibrium over with the poststrand passage one. These features distinguish echinoside A from other known Top2alpha inhibitors. As a result, echinoside A induced DNA double-strand breaks in a Top2-dependent manner. CONCLUSION: Echinoside A targets Top2alpha by unique interference with the binding of Top2 to DNA and by imparing the Top2-mediated DNA cleavage and religation, exerting potent in vitro and in vivo antitumor activities.

Novel mutations and phenotypes of epilepsy-associated genes in epileptic encephalopathies.

Epileptic encephalopathies are severe epilepsy disorders with strong genetic bases. We performed targeted next-generation sequencing (NGS) in 70 patients with epileptic encephalopathies. The likely pathogenicity of variants in candidate genes was evaluated by American College of Medical Genetics and Genomics (ACMG) scoring taken together with the accepted clinical presentation. Thirty-three candidate variants were detected after population filtration and computational prediction. According to ACMG, 21 candidate variants, including 18 de novo variants, were assessed to be pathogenic/likely pathogenic with clinical concordance. Twelve variants were initially assessed as uncertain significance by ACMG, among which 3 were considered causative and 3 others were considered possibly causative after analysis of clinical concordance. In total, 24 variants were identified as putatively causative, among which 19 were novel findings. SCN1A mutations were identified in 50% of patients with Dravet syndrome. TSC1/TSC2 mutations were detected in 66.7% of patients with tuberous sclerosis. STXBP1 mutations were the main findings in patients with West syndrome. Mutations in SCN2A, KCNT1, KCNQ2 and CLCN4 were identified in patients with epileptic infantile with migrating focal seizures; among them, KCNQ2 and CLCN4 were first identified as potential causative genes. Only one CHD2 mutation was detected in patients with Lennox-Gastaut syndrome. This study highlighted the utility of targeted NGS in genetic diagnoses of epileptic encephalopathies and a comprehensive evaluation of the pathogenicity of variants based on ACMG scoring and assessment of clinical concordance. Epileptic encephalopathies differ in genetic causes, and the genotype-phenotype correlations would provide insights into the underlying pathogenic mechanisms.

A triterpenoid saponin from Phytolacca esculenta.

A new triterpenoid saponin, 3-O-[beta-D-glucopyranosyl(1----4)-beta-D- xylopyranosyl]-28-O-beta-D-glucopyranosyl-30-methyloleanate-9(11), 12-dien- 2,3,23-trihydroxyl-28-oic acid, was isolated from the roots of Phytolacca esculenta. The structure was assigned by chemical methods and spectral analysis (1H, 13C, DEPT NMR, EIMS and FABMS) including 1H-1H COSY, 1H-13C COSY and 1H-1H NOESY.

A triterpenoid and its saponin from Phytolacca esculenta.

A new triterpenoid, esculentagenin, and its glycoside, esculentoside M, were isolated from the roots of Phytolacca esculenta and characterized as 11-oxo-3-O-methyloleanata-12-en-2 beta,3 beta,23-trihydroxy-28-oic acid and 3-O-[beta-D-glucopyranosyl(1----4)-beta-D-xylopyranosyl]-28-O-beta-D- glucopyranosyl-11-oxo-30-methyloleanate-12-en-2 beta,3 beta,23-trihydroxy-28-oic acid by spectral and chemical evidence.

[Studies on the structure of isoastilbin].

A new compound was isolated from Smilax glabra Roxb., named isoastilbin. It was identified as 5, 7, 3', 5'-tetrahydroxyl-flavanonol-3-O-alpha-L-rhamnopyranoside by means of chemical and spectrometric analysis (UV, IR, 1H-NMR, 13C-NMR, 2DNMR and FAB-MS).

[Studies on the chemical constituents of Uncaria yunanensis Hsia. C.C].

AIM: To research the chemical constituents from dried roots of Uncaria yunanensis Hsia. C. C. METHODS: Modern chromatography was used to isolate chemical components. Their structure were identified by spectral analysis. RESULTS: Seven compounds were isolated and identified as 3 beta, 6 beta, 19 alpha-trihydroxyurs-12-en-28 oic acid (I), 23-nor-24-esomethylene-3 beta, 6 beta-19 alpha-trihydroxyurs-12-en-28 oic acid (II), 3-oxo-6 beta, 19 alpha-dihydroxyurs-12-en-28 oic acid (III), oleanic acid (IV), 5,7,3',4'-tetrahydroxy-flavan-3-ol (V), beta-yohimbine (VI) and diangoutengjian I (VII). CONCLUSION: All of the above compounds were isolated for the first time from the root of this plant. Among them, compound VII is a new one.

[Studies on the triterpenoid saponins of the root bark of Aralia taibaiensis].

Four triterpenoid saponins were isolated from the root bark of Aralia taibaiensis Z. Z. Wang et H. C. Zheng. On the basis of their chemical properties and spectral data, they were identified as oleanolic acid-3-O-[beta-D-xylopyranosyl(1-->2)] [beta-D-glucopyranosyl(1-->3)]-beta-D-glucuronopyranoside (1), tarasaponin V (2), 3-O-¿beta-D-xylopyranosyl(1-->2)[beta-D-glucopyranosyl (1-->3)]-6'-O-ethyl-beta-D-glucuronopyranosyl¿-oleanolic acid-28-O-beta-D-glucopyranoside (3) and 3-O-¿beta-D-xylopyranosyl(1-->2) [beta-D-glucopyranosyl(1-->3)] -6'-O-butyl-beta-D-glucuronopyranosyl¿-oleanolic acid-28-O-beta-D-glucopyranoside (4). Compound 1 is a new natural product named taibaienoside VI. 2 was isolated from the title plant for the first time. 3 and 4 are new compounds and named taibaienoside VII and taibaienoside VIII, respectively.

[Studies on triterpenoids and their glycosides from Aralia dasyphylla Miq].

The structures of two triterpenoids and their glycosides were isolated from Aralia dasyphylla Miq. Their structures have been identified to be oleanoic acid(I), 16 beta-hydroxy-18 beta-H-oleanoic acid(II), oleanoic acid-28-O-beta-D-glucopyranoside(III) and 16 beta-hydroxy-18 beta-H-oleanoic acid-28-O-beta-D-glucopyranoside(IV), respectively, mainly through interpretation of UV, IR, MS, 1H and 13CNMR, DEPT, HMQC and HMBC spectra data. The stereochemistry of II has been confirmed by NOESY. Pharmacological experiments showed that the total saponins exerted preventative effect on CCl4-induced liver injury of male mice and hypoglycemic effect on a model of alloxan-induced diabetes in rats.

[Oleanolic acid saponins from the root bark of Aralia taibaiensis].

Five oleanolic acid saponins were isolated from the root bark of Aralia taibaiensis Z.Z. Wang et H.C. Zheng. By spectroscopic and chemical methods, they were identified as araloside A (1), 3-O-[alpha-L-arabinofuranosyl(1-->4)-6'-O-n-butyl-beta-D- glucuronopyranosyl]-oleanolic acid-28-O-beta-D-glucopyranoside (2), 3-O-[alpha-L-arabinofuranosyl(1-->4)-6'-O-ethyl-beta-D- glucuronopyra-nosyl]-oleanolic acid-28-O-beta-D-glucopyranoside (3), stipuleanoside R2(4) and 3-O-(beta-D-glucopyranosyl(1-->3) [alpha-L-arabinofuranosyl(1-->4)]-6'-O-ethyl-beta-D- glucuronopyranosyl)-oleanolic acid-28-O-beta-D-glucopyranoside (5). Saponin 1 and 4 were isolated from the title plant for the first time. 2, 3 and 5 are new compounds and named taibaienoside I, taibaienoside II and taibaienoside III, respectively.

[Inhibitory effects of esculentoside A on mouse macrophages and antibody production].

Esculentoside A (EsA) is a kind of saponin isolated from Phytolacca esculenta Van Houtte. It was shown to significantly inhibit phagocytic activity, intracellular and extracellular production of interleukin-1 by thioglycollate primed murine peritoneal macrophages in vitro at the concentration 0.01-1.0 mumol.L-1. In vivo experiments showed that EsA 2.5-5 mg.kg-1 markedly decreased serum hemolysin concentration in sensitized mice challenged with sheep red blood cells. Inhibition of antibody production by B lymphocytes, phagocytosis and the production of inflammatory mediators by macrophages may partially explain the wide and strong anti-inflammatory effect of EsA.

[Triterpenoid saponins from Eclipta prostrata L].

AIM: To study the triterpenoid saponins in the Chinese traditional medicine Eclipta prostrata L.. METHODS: Column chromatography with silica gel and HPLC were employed for the isolation and purification. The molecular structures were determined on the basis of spectral analysis (IR, MS, 1HNMR, 13CNMR, HMQC and HMBC). RESULTS: Two new triterpenoid saponins, named eclalbasaponins XI (4) and XII (5), were obtained and their structures were elucidated as 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl]-16 alpha-ethoxy-olean-12-ene-28-oic acid-28-O-beta-D-glucopyranoside and 3-O-[(2-O-sulfuryl-beta-D-glucopyranosyl) (1-->2)-beta-D-glucopyranosyl]-echinocystic acid-28-O-beta-D-glucopyranoside, respectively, along with three known saponins, eclalbasaponins II (1), I (2) and III (3). CONCLUSION: Compounds 4 and 5 are new compounds, 1 and 5 induced morphological deformation of Pyricularia oryzae mycelia.

[Studies on the anthraquinones of Cassia siamea].

AIM: To study the anthraquinone constituents of the stem of Cassia siamea. METHODS: The compounds were isolated by chromatography on silica gel, MHPLC, and identified on the basis of spectral analysis including IR, EI-MS, FAB-MS, 1HNMR, 13CNMR and DEPT. RESULTS: Three compounds were isolated and identified as: chrysophanol (I), chrysophanol-1-O-beta-D-glucopyranoside (II) and 1-[(beta-D-glucopyranosyl-(1-->6)-0-beta-D-glucopyranosyl) oxy]-8-hydroxyl-3-methy-9,10-anthraquinone (III). CONCLUSION: III is a new compound, II was obtained from this plant for the first time.

The targeting and functions of miRNA-383 are mediated by FMRP during spermatogenesis.

Our previous studies have shown that microRNA-383 (miR-383) expression is downregulated in the testes of infertile men with maturation arrest (MA). Abnormal testicular miR-383 expression may potentiate the connections between male infertility and testicular germ cell tumors. However, the mechanisms underlying the targeting and functions of miR-383 during spermatogenesis remain unknown. In this study, we found that fragile X mental retardation protein (FMRP) was associated with 88 miRNAs in mouse testis including miR-383. Knockdown of FMRP in NTERA-2 (NT2) (testicular embryonal carcinoma) cells enhanced miR-383-induced suppression of cell proliferation by decreasing the interaction between FMRP and miR-383, and then affecting miR-383 binding to the 3'-untranslated region of its target genes, including interferon regulatory factor-1 (IRF1) and Cyclin D1 both in vivo and in vitro. On the other hand, FMRP levels were also downregulated by overexpression of miR-383 in NT2 cells and GC1 (spermatogonia germ cell line). miR-383 targeted to Cyclin D1 directly, and then inhibited its downstream effectors, including phosphorylated pRb and E2F1, which ultimately resulted in decreased FMRP expression. Reduced miR-383 expression, dysregulated cyclin-dependent kinase 4 expression (one of the downstream genes of miR-383) and increased DNA damage were also observed in the testes of Fmr1 knockout mice and of MA patients with a downregulation of FMRP. A potential feedback loop between FMRP and miR-383 during spermatogenesis is proposed, and FMRP acts as a negative regulator of miR-383 functions. Our data also indicate that dysregulation of the FMRP-miR-383 pathway may partially contribute to human spermatogenic failure with MA.

Novel PRRT2 mutations in paroxysmal dyskinesia patients with variant inheritance and phenotypes.

Paroxysmal dyskinesias (PDs) are a group of episodic movement disorders with marked variability in clinical manifestation and potential association with epilepsy. PRRT2 has been identified as a causative gene for PDs, but the phenotypes and inheritance patterns of PRRT2 mutations need further clarification. In this study, 10 familial and 21 sporadic cases with PDs and PDs-related phenotypes were collected. Genomic DNA was screened for PRRT2 mutations by direct sequencing. Seven PRRT2 mutations were identified in nine (90.0%) familial cases and in six (28.6%) sporadic cases. Five mutations are novel: two missense mutations (c.647C>G/p.Pro216Arg and c.872C>T/p.Ala291Val) and three truncating mutations (c.117delA/p.Val41TyrfsX49, c.510dupT/p.Leu171SerfsX3 and c.579dupA/p.Glu194ArgfsX6). Autosomal dominant inheritance with incomplete penetrance was observed in most of the familial cases. In the sporadic cases, inheritance was heterogeneous including recessive inheritance with compound heterozygous mutations, inherited mutations with incomplete parental penetrance and de novo mutation. Variant phenotypes associated with PRRT2 mutations, found in 36.0% of the affected cases, included febrile convulsions, epilepsy, infantile non-convulsive seizures (INCS) and nocturnal convulsions (NC). All patients with INCS or NC, not reported previously, displayed abnormalities on electroencephalogram (EEG). No EEG abnormalities were recorded in patients with classical infantile convulsions and paroxysmal choreoathetosis (ICCA)/paroxysmal kinesigenic dyskinesia (PKD). Our study further confirms that PRRT2 mutations are the most common cause of familial PDs, displaying both dominant and recessive inheritance. Epilepsy may occasionally occur in ICCA/PKD patients with PRRT2 mutations. Variant phenotypes INCS or NC differ from classical ICCA/PKD clinically and electroencephalographically. They have some similarities with, but not identical to epilepsy, possibly represent an overlap between ICCA/PKD and epilepsy.

Three-dimensional characterization of resorption cavity size and location in human vertebral trabecular bone.

The number and size of resorption cavities in cancellous bone are believed to influence rates of bone loss, local tissue stress and strain and potentially whole bone strength. Traditional two-dimensional approaches to measuring resorption cavities in cancellous bone report the percent of the bone surface covered by cavities or osteoclasts, but cannot measure cavity number or size. Here we use three-dimensional imaging (voxel size 0.7×0.7×5.0 µm) to characterize resorption cavity location, number and size in human vertebral cancellous bone from nine elderly donors (7 male, 2 female, ages 47-80 years). Cavities were 30.10 ± 8.56 µm in maximum depth, 80.60 ± 22.23∗10(3) µm(2) in surface area and 614.16 ± 311.93∗10(3) µm(3) in volume (mean ± SD). The average number of cavities per unit tissue volume (N.Cv/TV) was 1.25 ± 0.77 mm(-3). The ratio of maximum cavity depth to local trabecular thickness was 30.46 ± 7.03% and maximum cavity depth was greater on thicker trabeculae (p<0.05, r(2)=0.14). Half of the resorption cavities were located entirely on nodes (the intersection of two or more trabeculae) within the trabecular structure. Cavities that were not entirely on nodes were predominately on plate-like trabeculae oriented in the cranial-caudal (longitudinal) direction. Cavities on plate-like trabeculae were larger in maximum cavity depth, cavity surface area and cavity volume than cavities on rod-like trabeculae (p<0.05). We conclude from these findings that cavity size and location are related to local trabecular microarchitecture.

Mosaic SCN1A mutations in familial partial epilepsy with antecedent febrile seizures.

SCN1A is the most relevant epilepsy gene. Mutations of SCN1A generate phenotypes ranging from the extremely severe form of Dravet syndrome (DS) to a mild form of generalized epilepsy with febrile seizures plus (GEFS+). Mosaic SCN1A mutations have been identified in rare familial DS. It is suspected that mosaic mutations of SCN1A may cause other types of familial epilepsies with febrile seizures (FS), which are more common clinically. Thus, we screened SCN1A mutations in 13 families with partial epilepsy with antecedent febrile seizures (PEFS+) using denaturing high-performance liquid chromatography and sequencing. The level of mosaicism was further quantified by pyrosequencing. Two missense SCN1A mutations with mosaic origin were identified in two unrelated families, accounting for 15.4% (2/13) of the PEFS+ families tested. One of the mosaic carriers with ~25.0% mutation of c.5768A>G/p.Q1923R had experienced simple FS; another with ~12.5% mutation of c.4847T>C/p.I1616T was asymptomatic. Their heterozygous children had PEFS+. Recurrent transmission occurred in both families, as noted in most of the families with germline mosaicism reported previously. The two mosaic mutations identified in this study are less destructive missense, compared with the more destructive truncating and splice-site mutations identified in the majority of previous studies. This is the first report of mosaic SCN1A mutations in families with probands that do not exhibit DS, but manifest only a milder phenotype. Therefore, such families with mild cases should be approached with caution in genetic counseling and the possibility of mosaicism origin associated with high recurrence risk should be excluded.