[Clinical analysis of 15 pediatric patients with tuberous sclerosis complex complicated by cardiac rhabdomyomas].

OBJECTIVE: To investigate the clinical features in children with tuberous sclerosis complex (TSC)-associated cardiac rhabdomyomas (CRM). METHODS: The clinical data of 15 children with TSC complicated by CRM were collected. The clinical features of the patients were analyzed, and TSC gene mutations were detected. RESULTS: Eleven cases (73%) developed multiple CRM. The majority of the tumors were located in the left and right ventricles. Most tumors presented as a round-like hyperechogenic mass with a clear margin on echocardiography. Arrhythmias occurred in 3 patients and 2 patients experienced heart failure. Gene mutation tests were performed in 2 patients, and pathogenic mutations were detected in both patients, which were TSC1 mutation and TSC2 mutation, respectively. Three patients were followed up for 6 to 38 months, and their CRM shrank or regressed spontaneously. CONCLUSIONS: TSC-associated CRM is generally multiple. Heart failure and arrhythmias may occur in some patients. Echocardiography is important for diagnosis of CRM. TSC-associated CRM has an inclination to spontaneous regression. TSC can be diagnosed at a molecular genetic level by TSC gene mutation detection.

Arabidopsis COBRA-LIKE 10, a GPI-anchored protein, mediates directional growth of pollen tubes.

Successful reproduction of flowering plants requires constant communication between female tissues and growing pollen tubes. Female cells secrete molecules and peptides as nutrients or guidance cues for fast and directional tube growth, which is executed by dynamic changes of intracellular activities within pollen tubes. Compared with the extensive interest in female cues and intracellular activities of pollen tubes, how female cues are sensed and interpreted intracellularly in pollen is poorly understood. We show here that COBL10, a glycosylphosphatidylinositol (GPI)-anchored protein, is one component of this pollen tube internal machinery. Mutations in COBL10 caused gametophytic male sterility due to reduced pollen tube growth and compromised directional sensing in the female transmitting tract. Deposition of the apical pectin cap and cellulose microfibrils was disrupted in cobl10 pollen tubes. Pollen tube localization of COBL10 at the apical plasma membrane is critical for its function and relies on proper GPI processing and its C-terminal hydrophobic residues. GPI-anchored proteins are widespread cell sensors in mammals, especially during egg-sperm communication. Our results that COBL10 is critical for directional growth of pollen tubes suggest that they play critical roles in cell-cell communications in plants.

HAPLESS13, the Arabidopsis µ1 adaptin, is essential for protein sorting at the trans-Golgi network/early endosome.

In plant cells, secretory and endocytic routes intersect at the trans-Golgi network (TGN)/early endosome (EE), where cargos are further sorted correctly and in a timely manner. Cargo sorting is essential for plant survival and therefore necessitates complex molecular machinery. Adaptor proteins (APs) play key roles in this process by recruiting coat proteins and selecting cargos for different vesicle carriers. The µ1 subunit of AP-1 in Arabidopsis (Arabidopsis thaliana) was recently identified at the TGN/EE and shown to be essential for cytokinesis. However, little was known about other cellular activities affected by mutations in AP-1 or the developmental consequences of such mutations. We report here that HAPLESS13 (HAP13), the Arabidopsis µ1 adaptin, is essential for protein sorting at the TGN/EE. Functional loss of HAP13 displayed pleiotropic developmental defects, some of which were suggestive of disrupted auxin signaling. Consistent with this, the asymmetric localization of PIN-FORMED2 (PIN2), an auxin transporter, was compromised in the mutant. In addition, cell morphogenesis was disrupted. We further demonstrate that HAP13 is critical for brefeldin A-sensitive but wortmannin-insensitive post-Golgi trafficking. Our results show that HAP13 is a key link in the sophisticated trafficking network in plant cells.

Arabidopsis RopGEF4 and RopGEF10 are important for FERONIA-mediated developmental but not environmental regulation of root hair growth.

We investigated a genetic pathway in root hair development in Arabidopsis thaliana, involving the receptor-like kinase FERONIA (FER), two guanine nucleotide exchange factors for ROPs (RopGEF4 and RopGEF10), and the small GTPase Rho of plants (ROPs). Loss- and gain-of-function analyses demonstrated distinct roles of RopGEF4 and RopGEF10 such that RopGEF4 is only important for root hair elongation, while RopGEF10 mainly contributes to root hair initiation. Domain dissection by truncation and domain-swapping experiments indicated that their functional distinctions were mainly contributed by the noncatalytic domains. Using fluorescent ratio imaging, we showed that functional loss of RopGEF4 and RopGEF10 additively reduced reactive oxygen species (ROS) production. Bimolecular fluorescence complementation experiments demonstrated that RopGEF4 and RopGEF10 had the same interaction specificity as ROPs, suggesting common downstream components. We further showed that the promoting effects of environmental cues such as exogenous auxin and phosphate limitation on root hair development depended on FER. However, although functional loss of RopGEF4 and RopGEF10 largely abolished FER-induced ROS production, it did not compromise the responses to FER-mediated environmental cues on root hair development. Our results demonstrated that RopGEF4 and RopGEF10 are genetic components in FER-mediated, developmentally (but not environmentally) regulated, root hair growth.

Functional characterization of a KCNAB3 genetic epilepsy with febrile seizures plus adult mouse model.

Background: Genetic epilepsy with febrile seizures plus (GEFS+) is a type of epileptic syndrome closely related to heredity factors, which can be caused by gene mutations. However, it still remains unclear how these mutations result in seizures. Previously, we identified a new heterozygous missense mutation of the KCNAB3 gene, H258R, in the GEFS+ family; the electric currents of the human embryonic kidney 293 (HEK293) cells co-expressing Kvß3 (H258R) and Kv1.1 showed obvious inactivation. This study sought to examine the effects of this mutation on the potassium channels in the mammalian brain. Methods: Mutant mice were generated by introducing the human H258R missense mutation within exon 10 at an equivalent position in the mouse KCNAB3 gene via CRISPR/Cas9 and homologous recombination. A patch clamp was used to detect the potassium currents in the pyramidal cells of the hippocampal CA1 region of the mutant mice. The total potassium currents of the pyramidal cells in the hippocampal CA1 region of KCNAB3 [wild-type (WT)] and KCNAB3 (H258R) adult mice were recorded with increased voltage. Results: We found a decreased total potassium current in the H258R group but no significant differences at a maximum voltage (+80 mV; P>0.05). Conclusions: These results suggest that the KCNAB3 mutation reduced hippocampal potassium currents in this mouse model.

Anti-inflammatory role of IL-17 in experimental autoimmune uveitis.

Previous studies have shown that IL-17 is a strong proinflammatory cytokine and that IL-17-producing autoreactive T cells play a major role in the pathogenesis of autoimmune diseases. In a previous study, we showed that injection of experimental autoimmune uveitis-susceptible mice with anti-IL-17 Abs blocked subsequent disease development. To determine whether administration of IL-17 to experimental autoimmune uveitis-susceptible Lewis rats and B10RIII mice injected with disease-inducing peptides enhanced disease susceptibility, we injected the recipient animals with various doses of human rIL-17 (hIL-17). Unexpectedly, the treated animals showed significant amelioration of disease; in addition, both the intensity of the autoreactive response and cytokine production by the autoreactive T cells induced by immunization with uveitogenic peptides were significantly decreased. Our results show that IL-17 has anti-inflammatory activity and that this cytokine can suppress the development of autoimmune disease.

Protective effect of histatin 1 against ultraviolet-induced damage to human corneal epithelial cells.

The aim of the present study was to investigate the role of histatin 1 (Hst1) in human corneal epithelial cells (HCECs) exposed to ultraviolet (UV) radiation. Prior to UV irradiation for various durations, HCECs were pre-treated with different concentrations of Hst1 and the effect on cell apoptosis and cell viability were examined by flow cytometry, alamarBlue® and MTT assays to determine the optimal concentration of Hst1 and UV dose. Cells were then subjected to quantitative PCR, ELISA and western blot analysis to determine the expression of cell damage-associated genes. HCECs exposed to UV light for 1 h displayed decreased viability when compared to that of control cells, and a 3 h UV exposure markedly increased the apoptotic rate of HECEs, while apoptosis was inhibited by pre-treatment with Hst1. UV radiation downregulated expression of insulin-like growth factor (IGF)-1 and B-cell lymphoma 2 (Bcl-2), while it upregulated Bcl-2-associated X protein (Bax) expression. Hst1 protected HCECs against UV-induced damage by upregulating the expression of IGF-1 protein and increasing the Bcl-2/Bax ratio. In conclusion, Hst1 may prevent UV-induced damage to corneal epithelial tissue injury and promote its healing.

Degradation of MTBE and TBA by a new isolate from MTBE-contaminated soil.

Methyl tert-butyl ether (MTBE), a gasoline additive, possesses serious problems to the environmental health. In the present study, a bacterial culture named A-3 which could effectively degrade MTBE was isolated from the MTBE contaminated soil. The isolate was identified as Chryseobacterium sp., a new species capable of degrading MTBE. In order to enhance its degradation ability, selected environment factors were investigated. The results showed that the optimal temperature was in the range of 25-30 degrees C, the pH was 7.0, the inoculum size was 2 x 10(8) CFU/ml and the optimal concentration of MTBE was from 50 to 100 mg/L. The maximum MTBE utilization rate (upsilon(max)) was 102 nmol MTBE/(mg cell protein x h). Furthermore, it was found that the isolate could also degrade tert-butyl alcohol (TBA). The degradation rates of TBA were much faster than those of MTBE. The additional TBA would lead to the decrease of the initial MTBE degradation rate and the inhibitory effect of TBA increased with the increase of TBA concentration. Similar protein profiles at least seven peptides were demonstrated after SDS-PAGE analysis of crude extracts obtained from the cells growing in MTBE and TBA culture.

Characterization and distribution of polycyclic aromatic hydrocarbon in sediments of Haihe River, Tianjin, China.

In this study sediment samples were collected from 13 sites of Haihe River in Tianjin City, China, sixteen of priority polycyclic aromatic hydrocarbons (PAHs) listed in USEPA were analyzed by means of GC-MS. The total concentrations of PAH ranged from 774.81 to 255371.91 ng/g dw, and two to four rings of PAHs were dominant in sediment samples. Molecular ratios, such as phenanthrene/anthracene, fluoranthene/pyrene and low-molecular-weight PAH /high-molecular-weight PAH, were used to study the possible sources of pollution. It indicated a mixed pattern of parolytic and petrogenic inputs of PAHs in sediments in Haihe River. The petrogenic PAHs may be mainly derived from the leakage of refined products, e.g., gasoline, diesel fuel and fuel oil vehicle traffics or gas stations from urban area. The pyrolytic PAHs might be from the discharge of industrial wastewater and the emission of atmospheric particles from petrochemical factories. In addition, the levels of PAHs in the urban and industrial areas are far beyond the values reported from other rivers and marine systems reported. This situation may be due to polluted discharging from some petrochemical industrial manufactories and worse traffic conditions in Tianjin.

Laboratory column study for remediation of MTBE-contaminated groundwater using a biological two-layer permeable barrier.

In this study, an in situ biological two-layer permeable reactive barrier system consisting of an oxygen-releasing material layer followed by a biodegradation layer was designed to evaluate the remediation effectiveness of MTBE-contaminated groundwater. The first layer containing calcium peroxide (CaO(2)) and other inorganic salts is to provide oxygen and nutrients for the immobilized microbes in the second layer in order to keep them in aerobic condition and maintain their normal metabolism. Furthermore, inorganic salts such as potassium dihydrogen phosphate (KH(2)PO(4)) and ammonium sulphate ((NH(4))(2)SO(4)) can also decrease the high pH caused by the alkali salt degraded from CaO(2). The second layer using granular expanded perlite as microbial carrier is able to biodegrade MTBE entering the barrier system. Batch experiments were conducted to identify the appropriate components of oxygen-releasing materials and the optimum pH value for the biodegradation of MTBE. At pH=8.0, the biodegradation efficiency of MTBE is the maximum and approximately 48.9%. A laboratory-scale experiment using two continuous upflow stainless-steel columns was then performed to evaluate the feasibility of this designed system. The fist column was filled with oxygen-releasing materials at certain ratio by weight. The second column was filled with expanded perlite granules immobilizing MTBE-degrading microbial consortium. Simulated MTBE-contaminated groundwater, in which dissolved oxygen (DO) content was 0mg/L, was pumped into this system at a flow rate of 500mL/d. Samples from the second column were analyzed for MTBE and its major degradation byproduct. Results showed that MTBE could be removed, and its metabolic intermediate, tert-butyl alcohol (TBA), could also be further degraded in this passive system.

Groundwater remediation engineering sparging using acetylene--study on the flow distribution of air.

Air sparging (AS) is an emerging method to remove VOCs from saturated soils and groundwater. Air sparging performance highly depends on the air distribution resulting in the aquifer. In order to study gas flow characterization, a two-dimensional experimental chamber was designed and installed. In addition, the method by using acetylene as the tracer to directly image the gas distribution results of AS process has been put forward. Experiments were performed with different injected gas flow rates. The gas flow patterns were found to depend significantly on the injected gas flow rate, and the characterization of gas flow distributions in porous media was very different from the acetylene tracing study. Lower and higher gas flow rates generally yield more irregular in shape and less effective gas distributions.

Huangzhi Oral Liquid Prevents Arrhythmias by Upregulating Caspase-3 and Apoptosis Network Proteins in Myocardial Ischemia-Reperfusion Injury in Rats.

To study the effect of Huangzhi oral liquid (HZOL) on I/R after 2 h and 4 h and determine its regulatory function on caspase-3 and protein networks. 70 SD male rats were randomly divided into seven groups and established myocardial I/R injury model by ligating the left anterior descending coronary artery. Myocardial infarction model was defined by TTC staining and color of the heart. The levels of CK-MB, CTnI, C-RPL, SOD, and MDA were tested at 2 h and 4 h after reperfusion. HE staining and ultramicrostructural were used to observe the pathological changes. The apoptotic index (AI) of cardiomyocyte was marked by TUNEL. The expression levels of caspase-3, p53, fas, Bcl-2, and Bax were tested by immunohistochemistry and western blot. HZOL corrected arrhythmia, improved the pathologic abnormalities, decreased CK-MB, CTnI, C-RPL, MDA, AI, caspase-3, p53, fas, and Bax, and increased SOD ans Bcl-2 with different times of myocardial reperfusion; this result was similar to the ISMOC (P > 0.05). HZOL could inhibit arrhythmia at 2 and 4 h after I/R and ameliorate cardiac function, which was more significant at 4 h after reperfusion. This result may be related to decreased expression of caspase-3, p53, and fas and increased Bcl-2/Bax ratio.

[Relationship between interleukin-6 gene-572C/G polymorphism and chronic periodontitis].

OBJECTIVE: To investigate the relationship between interleukin-6 (IL-6) gene -572C/G polymorphism and the risk of chronic periodontitis disease. METHODS: IL-6-572C/G genotype polymorphism was examined in 93 patients with chronic periodontitis and 96 control subjects by PCR restriction fragment length polymorphism (PCR-RFLP) method. RESULTS: IL-6-572GG genotype and G allele were more frequent in chronic periodontitis patients than that in control subjects (P<0.05). -572CC, CG and GG genotype frequency were 52.7 % , 40.9 %, 6.4 % in chronic periodontitis patients and 67.7 %, 31.3%, 1.0 % in control subjects, respectively. -572CG + GG genotype and G allele were more frequent in patients than that in control subjects (P<0.05). Compared with CC genotype,the odds ratio for chronic periodontitis was 1.88 (95% CI: 1.04-3.40, P<0.05) for CG + GG genotype. CONCLUSIONS: The frequency of -572CG + GG genotype was higher in patients with chronic periodontitis compared with the control subjects. IL-6-572C/G polymorphism may be a genetic susceptibility factor for chronic periodontitis in Chinese Hans population.