[Clinical and molecular genetic analysis of Angelman syndrome with oculocutaneous albinism type 2: A case report and literature review].

To summarize the clinical diagnosis and treatment process and genetic test results and characteristics of one child with Angelman syndrome (AS) complicated with oculocutaneous albinism type 2 (OCA2), and to review the literature. "Angelman syndrome" "P gene" and "Oculocutaneous albinism type 2" were used as keywords to search at CNKI, Wanfang, and PubMed databases (from creation to December 2019). Then all the patients were analyzed. The patient in this study was a girl aged 1 year. After birth, she was found to present as white body, yellow hair, and nystagmus. She could raise her head at the age of 2 months and turn over at the age of 7 months. The head circumference was 42 cm and she could not sit alone or speak at present. Trio-based exome sequencing revealed that the patient carried a homozygous mutation of c.168del (p.Gln58ArgfsTer44) in the P gene, and her father was heterozygous and her mother was wild-type. The detection of copy number variation showed deletion on the maternal chromosome at 15q11.2-13.1 region (P gene located in this region) in the patient. Until December 2019, a total of 4 cases in the 4 literature had been reported. Adding our case here, the 5 cases were summarized and found that all the cases showed white skin, golden hair, and shallow iris after birth. Comprehensive developmental delay was found around 6 months of age after birth, and the language remained undeveloped in 2 cases till follow-up into childhood. Seizures occurred in 4 patients. Two cases had ataxia. All the 5 cases had acquired microcephaly. Two cases had a family history of albinism. Electroencephalogram monitoring was completed in 3 cases and the results were abnormal. Genetic tests showed that all the 5 cases had deletion on maternal chromosome at 15q11-13 region. Four cases carried mutation of P gene on paternal chromosome. And 1 case was clinically diagnosed as OCA2 without P gene test. AS combined with OCA2 is relatively rare. OCA2 is easily diagnosed based on the obvious clinical manifestations after birth. When combined with clinical manifestations such as neurodevelopmental delay, it might indicate the possibility of AS that is hardly diagnosed clinically at an early stage. Genetic tests can reveal the cross-genetic phenomenon of AS and OCA2 and the complex of them can be eventually diagnosed.

[Clinical features of IgG4-related lung disease].

Objective: To explore the clinical features of IgG4-related lung disease. Methods: The clinical data of 60 patients diagnosed with IgG4-related lung disease in Peking University People's Hospital from February 2012 to May 2021 were retrospectively collected. Analysis was made to explore the features of clinical manifestation, laboratory, imaging, prognosis and other characteristics of the disease. Results: A total of 60 patients were included, with 40 males, age of (58.2±12.9) years, an age of onset of (57.1±13.2) years, and 31.7% (19 cases) of the patients had a history of allergic disease. 36.7% (22 cases) of the patients had respiratory symptoms during the disease. 94.6% (53/56) of patients had serum IgG4>1.35 g/L, 24.1% (14/58) of patients had increased eosinophils, 79.2% (38/48) of patients had increased IgE level, and 53.7% (29/54) of patients had decreased C3 or C4. Common imaging findings included nodular changes (38 cases, 63.3%), mediastinal and/or hilar lymphadenopathy (34 cases, 56.7%), and ground glass opacities (31 cases, 51.7%). Fifty-three cases (88.3%) showed two or more imaging changes. The pathological examination of the patient was mainly characterized by lymphoplasmacytic infiltration and fibrosis, with only one case of phlebitis obliterans. Compared with the asymptomatic group (38 cases), patients with respiratory symptoms (22 cases) showed higher level of serum total IgG and eosinophils (43.2 vs 17.8 g/L, 0.30×109/L vs 0.14×109/L, P<0.05), lower proportion of allergic diseases, and higher proportion of consolidation shadows on chest CT (P<0.05). There were no significant differences in serum IgG4, IgE, complement levels, and imaging outcomes after treatment between the two groups (P>0.05). Conclusions: The clinical manifestations of IgG4-related lung disease are atypical, and asymptomatic patients account for a high proportion. The imaging of the disease is highly heterogeneous, and patients are prone to show coexisted multiple imaging changes. The main clinical features and imaging outcomes of patients with and without respiratory symptoms are not significantly different.

[Clinical efficacy observation of omalizumab on patients with moderate to severe allergic asthma for one year].

To observe the symptom control, pulmonary function changes and safety of use of omalizumab in patients with moderate to severe allergic asthma for 1 year. A small sample self-controlled study before and after treatment was conducted to retrospective analysis involved 17 patients with moderate to severe asthma who received omalizumab therapy for 12 months in Peking University People's Hospital and Beijing Jishuitan Hospital from January 2020 to December 2021. The clinical symptoms and pulmonary function changes were compared before treatment, after 6 months and 12 months of treatment, and the clinical data such as the use of other drugs and adverse reactions were observed. Statistical data are collected using the median method, and non-parametric paired Wilcoxon analysis was used for pairwise comparison. Before treatment with omalizumab, the patients' FeNO value was 79(58, 121) ppb, and the total serum IgE was 228(150.5, 345.5) IU/ml. After 6 months of omalizumab therapy, the percent predicted value of the forced expiratory volume in 1 second (FEV1%) before inhaled bronchodilator increased from 86.70(82.65, 91.35)% to 90.90(87.70, 95.85)% (Z=-3.626, P<0.001). The FEV1%pred after inhaled bronchodilator increased from 92.60(85.75, 96.90)% to 94.30(89.95, 98.15)% (Z=-2.178, P=0.029). The absolute value of improvement in FEV1 decreased from 150(95, 210)ml to 50(20, 125) ml (Z=-2.796, P=0.005), and the improvement rate decreased from 6.60(3.80, 7.85)% to 1.90(0.75, 4.85)% (Z=-2.922, P=0.003). After 12 months of treatment, the FEV1%pred before inhaled bronchodilator further increased to 92.90 (91.60, 98.15)% (Z=-3.575, -2.818, and P<0.001, 0.005 compared with before treatment and 6 months after treatment, respectively). The FEV1%pred after inhaled bronchodilator increased to 96.80 (91.90, 101.25)% (Z=-3.622, -1.638, and P<0.001, 0.008 compared with before treatment and after 6 months of treatment, respectively). The absolute value of improvement in FEV1 was 70 (35, 120) ml (P=0.004, 0.842 before treatment and 6 months after treatment, respectively), and the improvement rate was 3.0(1.0, 5.0)% (Z=-2.960, -0.166, and P=0.003, 0.868, compared with before treatment and after 6 months of treatment, respectively). After 12 months of treatment, ACT increased from 13 (10.5, 18) before treatment to 24 (23, 25) (Z=-3.626,P<0.001). Only 1 patient experienced an injection site skin reaction during treatment. Therefore, after 6 months and 12 months of treatment with omalizumab, the patient's lung function improved and symptoms were relieved, which could effectively prevent the acute exacerbation of asthma. Omalizumab treatment is safe and well tolerated, and no effect on blood pressure and blood glucose was observed.

Nomogram predicting early neurological improvement in ischaemic stroke patients treated with endovascular thrombectomy.

BACKGROUND AND PURPOSE: Early neurological improvement (ENI) after endovascular thrombectomy (EVT) has been associated with favorable outcomes. This study aimed to identify the optimal definition of ENI and develop a nomogram for predicting ENI after EVT in acute ischaemic stroke. METHODS: Patients with EVT were enrolled from a multicenter registry as the training cohort. The receiver operating characteristic curve was used to estimate the optimal threshold for ENI at 24 h of EVT. Logistic regression analysis was utilized to generate the best-fit nomogram for predicting ENI. The discrimination of the nomogram was assessed using the area under the receiver operating characteristic curve (AUC). An additional 447 patients from two stroke centers were prospectively recruited as the test cohort for validating the nomogram. RESULTS: A total of 612 patients with EVT were included in the training cohort. The optimal threshold for predicting 3-month favorable outcome (modified Rankin Scale 0-2) was an improvement of the National Institutes of Health Stroke Scale (NIHSS) score by ≥6 points (AUC 0.875; sensitivity 79.5%; specificity 90.7%). Age, blood glucose, recanalization, symptomatic intracranial hemorrhage (sICH) and baseline Alberta Stroke Program Early Computed Tomography Score (ASPECTS) were independently associated with ENI, and were incorporated in the nomogram. The AUC of the nomogram was 0.795 in the training cohort and 0.752 in the test cohort. CONCLUSIONS: A reduction of NIHSS score ≥6 appeared to be the optimal definition of ENI. The nomogram composed of age, blood glucose, recanalization, sICH and baseline ASPECTS may predict the probability of ENI in ischaemic stroke patients treated with EVT.

Potential probiotics Lactobacillus casei K11 combined with plant extracts reduce markers of type 2 diabetes mellitus in mice.

AIMS: Probiotics and plant extracts have been used to prevent the development of type 2 diabetes mellitus (T2DM). The study aimed to explore the effect of the interaction between potential probiotics and bitter gourd extract (BGE) or mulberry leaf extract (MLE) on T2DM. METHODS AND RESULTS: Potential probiotics were tested for their gastrointestinal tract viability and growth situation combined with BGE and MLE in vitro. The diabetes model was constructed in C57BL/6 mice, and the potential effect and mechanism of regulating blood glucose were verified. Hematoxylin-eosin staining (HE), gas chromatography (GC), ELISA, and RT-PCR were also used for analysis. The results showed that Lactobacillus casei K11 had outstanding gastrointestinal tract viability and growth situation with plant extracts. Administration of L. casei K11 combined with BGE and MLE significantly reduced blood glucose levels and ameliorated insulin resistance in diabetic mice than the administration of Lactobacillus paracasei J5 combined with BGE and MLE. Moreover, in L. casei K11 combined with BGE and MLE groups, lipid metabolism, oxidative stress, and proinflammatory cytokine levels were regulated. Furthermore, the results indicated that L. casei K11 combined with BGE and MLE improved free fatty acid receptor 2 (FFAR2) upregulation, glucagon-like peptide-1 (GLP-1) secretion, and short-chain fatty acid (SCFA) levels. CONCLUSIONS: These findings showed that L. casei K11 combined with BGE and MLE modified the SCFA-FFAR2-GLP-1 pathway to improve T2DM. SIGNIFICANCE AND IMPACT OF THE STUDY: This study identified a new modality for evaluating interactions between potential probiotics and plant extracts. Our findings revealed that L. casei K11 combined with BGE and MLE significantly promoted the SCFA-FFAR2-GLP-1 pathway to inhibit T2DM.

Breast milk flora plays an important role in infantile eczema: cohort study in Northeast China.

AIMS: Infantile eczema, usually coupled with a range of hypersensitive phenotypes, has come into notice with its rising prevalence and unclear pathogenesis. Recent studies show close ties between eczema and an infant's intestinal flora. To gain a further understanding of the interactions between microbiota and eczema, we studied the breast milk flora as a new factor and present the links among breast milk flora, infant intestinal flora and infantile eczema through a cohort study in Northeast China. METHODS AND RESULTS: Fifty-two families were recruited with either an eczema or healthy infant younger than 6 months. Analysis and predictions using amplicon sequencing of microbiota found that Bifidobacterium and Bacteroidetes were enriched in healthy and eczema infant stools, respectively, consistent with previous reports. For breast milk flora, more 'positive' bacteria such as Akkermansia were enriched in breast milk from healthy infants' mothers. Further, higher bacterial delivery efficiencies were found in pairs of breast milk flora and infants' stool flora of families with eczema infants compared with families with healthy infants. Bacteroidetes, a widely known indicator of eczema, was found delivered more in eczema pairs. Further metagenomic predictions revealed that the breast milk microbiota participated significantly less in metabolism and immune system pathways, particularly in antigen processing and presentation and in Th17 cell-related pathways. CONCLUSIONS: In conclusion, as with other components of breast milk, the breast milk microbiota closely associates with infants' health via mother-infant bacterial delivery and metabolic functions. SIGNIFICANCE AND IMPACT OF THE STUDY: Our research aimed to fill the gap between the eczema and breast milk flora and describe the connections among breast milk and intestinal flora and eczema.

In vitro assays on the susceptibility of four species of nematophagous fungi to anthelmintics and chemical fungicides/antifungal drug.

Using nematophagous fungi for the biological control of animal parasitic nematodes will become one of the most promising strategies in the search for alternative chemical drugs. The purpose of this study was to check the in vitro activity of four anthelmintics, four chemical fungicides and two antifungal drugs on the spore germination of nematophagous fungi: Duddingtonia flagrans (SF170), Arthrobotrys oligospora (447), Arthrobotrys superba (435) and Arthrobotrys sp. (PS011). A modified 24-well cell culture plate assay was conducted to evaluate the susceptibility of nematophagous fungi against drugs tested by calculating the effective middle concentrations (EC50 ) of each tested drug to inhibit the germination of fungal spores. EC50 ranged between 0·7 and 47·2 µg ml-1 for fenbendazole, thiabendazole and ivermectin, except levamisole (546·5-4057·8 µg ml-1 ). EC50 of tested fungicides was 0·6-2·3 µg ml-1 for carbendazim, 55·9-247·4 µg ml-1 for metalaxyl, 24·4-45·2 µg ml-1 for difenoconazole, and 555·9-1438·3 µg ml-1 for pentachloronitrobenzene (PCNB). EC50 of two antifungal drugs was 0·03-3·4 µg ml-1 for amphotericin B and 0·3-10·9 µg ml-1 for ketoconazole. The results showed that 10 tested drugs, except for levamisole and PCNB, had in vitro inhibitory effects on nematophagous fungi. The chlamydospores of D. flagrans had the highest sensitivity to nine tested drugs, except for ketoconazole.

[Levels of sgRNA as a Major Factor Affecting CRISPRi Knockdown Efficiency in K562 Cells].

To determine how nuclease deactivated Cas9 (dCas9) or single-guide RNA (sgRNA) expression levels affect the knockdown efficiency of CRISPRi, we created K562 cell clones expressing KRAB-dCas9 protein either with the inducible Tet-on system or with the constitutive SFFV promotor. Single clones were selected by fluorescence-activated cell sorting (FACS) for further study. Six genes with various expression levels were targeted using lentiviral sgRNA from two libraries in four cell clones with various KRAB-dCas9 expression levels. The expression level of dCas9 protein/sgRNA levels and the knockdown efficiency were determined by flow cytometry. The cell clone with the highest KRAB-dCas9 expression level achieved effective CRISPRi knockdown. The data describing this clone were statistically different from that on other clones, indicating the strong KRAB-dCas9 expression might be a prerequisite for CRISPRi. By adopting different multiplicity of infection (MOI) in lentiviral transduction of this clone, we modified the expression level of sgRNA and found that the knockdown efficiency was neither affected by the target gene expression level nor correlated with KRAB-dCas9 levels, which remained relatively constant across all knockdown experiments (coefficient of variation = 2.2%). As an example, the following levels of the knockdowns: 74.72, 72.28 and 39.08% for mmadhc, rpia and znf148 genes, respectively, were achieved. These knockdown efficiencies correlated well with the respective sgRNA expression levels. Linear regression models built using this data indicate that the knockdown efficiency may be significantly affected by the levels of both KRAB-dCas9 and sgRNA. Notably, the sgRNA levels have greater impact, being a major factor affecting CRISPRi efficiency.

[Clinical study of dienogest in the treatment of refractory endometriosis-associated pain].

Objective: To evaluate the efficacy and safety of dienogest (DNG) in the treatment of refractory endometriosis-associated pain (REAP). Methods: In this study, REAP was defined according to the following criteria: (1) the pain duration was ≥12 months and visual analogue scale (VAS)≥60 mm; (2) the previous treatments with over two medicines like oral contraceptives and levonorgestrel-releasing intrauterine system failed to achieve satisfactory relief of pain, with VAS reduction less than 50%; with gonadotropin-releasing hormone agonist or mifepristone, the pain could be controlled temporarily, but it recurred after discontinuation of medicines; (3) the pain could not be relieved by surgery or even repeated surgeries. In the present study, 48 patients with REAP were treated with DNG 2 mg/day orally and the clinical outcomes were retrospectively analyzed. The VAS scores, levels of CA125, estradiol, FSH, LH and changes in the size of endometriotic lesions before and after treatment were compared respectively. The side effects were also analyzed. Results: The average duration of DNG treatment was (20.1±12.8) months. After 3 months of medication, the VAS score was significantly reduced from (77.9±15.8) mm to (20.8±10.7) mm (P<0.01), and CA125 level was significantly reduced from (95±139) kU/L to (38±45) kU/L (P<0.05). The effects were maintained with continuation of DNG treatment. Endometriotic lesions tended to shrink, after 12 months of DNG treatment, the size of ovarian endometriomas was reduced significantly from (3.1±1.0) cm to (1.9±1.2) cm (P<0.05). The mean level of estradiol was maintained at 124.82-221.04 pmol/L and levels of FSH and LH did not change significantly during the treatment. The major side effect was irregular bleeding (75%, 36/48). Conclusions: DNG could effectively relieve REAP and is a well-tolerated therapy. It may supply an alternative option for patients with REAP.

A novel nomogram to predict early neurological deterioration in patients with acute ischaemic stroke.

BACKGROUND AND PURPOSE: Acute ischaemic stroke (AIS) is a vital cause of mortality and morbidity in China. Many AIS patients develop early neurological deterioration (END). This study aimed to construct a nomogram to predict END in AIS patients. METHODS: Acute ischaemic stroke patients in Nanjing First Hospital were recruited as the training cohort. Additional patients in Nantong Third People's Hospital were enrolled as the validation cohort. Multivariate logistic regression was utilized to establish the nomogram. Discrimination and calibration performance of the nomogram were tested by concordance index and calibration plots. Decision curve analysis was employed to assess the utility of the nomogram. RESULTS: In all, 1889 and 818 patients were recruited in the training and validation cohorts, respectively. Age [odds ratio (OR) 1.075; 95% confidence interval (CI) 1.059-1.091], diabetes mellitus (OR 1.673; 95% CI 1.181-2.370), atrial fibrillation (OR 3.297; 95% CI 2.005-5.421), previous antiplatelet medication (OR 0.473; 95% CI 0.301-0.744), hyper-sensitive C-reactive protein (OR 1.049; 95% CI 1.036-1.063) and baseline National Institutes of Health Stroke Scale (OR 1.071; 95% CI 1.045-1.098) were associated with END and incorporated in the nomogram. The concordance index was 0.826 (95% CI 0.785-0.885) and 0.798 (95% CI 0.749-0.847) in the training and validation cohorts. By decision curve analysis, the model was relevant between thresholds of 0.06 and 0.90 in the training cohort and 0.08 and 0.77 in the validation cohort. CONCLUSIONS: The nomogram composed of hyper-sensitive C-reactive protein, age, diabetes mellitus, atrial fibrillation, previous antiplatelet medication and baseline National Institutes of Health Stroke Scale may predict the risk of END in AIS patients.

Profiles of gut microbiota in children with obesity from Harbin, China and screening of strains with anti-obesity ability in vitro and in vivo.

AIMS: The purpose of this study was to investigate the characteristics of gut microbiota of children with obesity in Harbin, China and to screen anti-obesity strains in vitro and in vivo. METHODS AND RESULTS: The gut microbiota of children with obesity and normal-weight children were investigated by high-throughput sequencing, and based on the different composition in gut microbiota, the strains with potential anti-obesity properties were screened in vitro and in vivo. Compared with normal-weight children, the Firmicutes to Bacteroidetes ratio in children with obesity decreased. Moreover the relative abundances of Lactobacillus and Bifidobacterium in children with obesity were decreased, while the relative abundance of Akkermansia increased. After a series of screening in vitro and in vivo, nine strains were found inhibiting the body weight gain of HFD-fed mice, of which two strains showed significant effects (P < 0·05). CONCLUSIONS: There were significant changes in gut microbiota of children with obesity from Harbin, China. The obtained strains showed obvious anti-obesity effects, and the screening methods used in this study were effective. SIGNIFICANCE AND IMPACT OF THE STUDY: This study enriched the research results on the characteristics of gut microbiota of children with obesity in different regions of the world. Moreover we established a new and effective method for screening anti-obesity strains, and obtained effective strains.

Epileptic negative myoclonus restricted to lower limbs in benign childhood focal epilepsy with vertex spikes.

BACKGROUND AND PURPOSE: The aim was to determine the electroclinical findings in benign childhood focal epilepsy with vertex spikes (BEVS) with epileptic negative myoclonus (ENM) restricted to the lower limbs. METHODS: The electroencephalogram database of Peking University First Hospital and medical records of patients with BEVS and ENM restricted to the lower limbs were reviewed. RESULTS: Twenty-seven patients with BEVS had ENM restricted to the lower limbs. Twelve started as ENM restricted to the lower limbs. The age at seizure onset ranged from 1.5 to 4.8 years. During the course, half of the 12 patients developed focal sensorimotor seizures and then were diagnosed as benign childhood epilepsy with centrotemporal spikes (BECTS), with BEVS (four cases) and without BEVS (two cases). Five of them had electrical status epilepticus during sleep (ESES) and met the diagnostic criteria of atypical benign partial epilepsy (ABPE). Fifteen of the 27 patients had ENM restricted to the lower limbs during the course. The age at seizure onset ranged from 1.3 to 9.8 years. All had ESES and were diagnosed as ABPE, 11 as ABPE with BEVS and four as ABPE evolving into BEVS. CONCLUSIONS: Benign childhood focal epilepsy with vertex spikes (BEVS) might represent a specific epileptic syndrome of the continuum of benign childhood focal epilepsy. ENM restricted to the lower limbs was a special phenomenon in BEVS. BEVS could overlap with BECTS or evolve into BECTS and further into ABPE and vice versa. Ignorance of vertex spikes with associated ENM restricted to the lower limbs might lead to a misunderstanding of BEVS, a specific type of benign childhood focal epilepsy.

[Photosensitive tonic-clonic seizures:a continuum between focal and generalized seizures].

OBJECTIVE: To investigate whether the tonic-clonic seizure (TCS) induced by intermittent photic stimulation (IPS)was generalized tonic-clonic seizure (GTCS)or partial secondarily tonic-clonic seizure (PGTCS),and to analyze the relationship between them. METHODS: Video-electroencephalogram (VEEG)database of Peking University First Hospital from March 2010 to October 2018 were reviewed. Fifteen cases with idiopathic epilepsy who had TCS induced by IPS were included in this study, and their clinical and electroencephalogram (EEG)characteristics were retrospectively analyzed. RESULTS: In this study, 4 of the 15 cases were boys and 11 were girls. The age of seizure onset ranged from 1 to 13 years. According to the medical records: 12 cases were considered as GTCS,while the remaining 3 cases were considered as PGTCS. The age at VEEG monitoring ranged from 2.5 to 16.0 years. All backgrounds of the VEEG were normal. Interictal discharges:generalized discharges in 11 cases, of which 4 cases coexisted with posterior discharges, 2 cases coexisted with Rolandic discharges, the other 5 cases merely had generalized discharges; merely focal discharges in two cases, one in the Rolandic area and the other in the posterior area; no interictal discharge in the remaining 2 cases. IPS induced photoparoxysmal response (PPR)results: 2 cases without PPR,the remaining 13 cases with PPR of generalized discharges, and 6 of the 13 cases coexisted with posterior discharges. IPS induced photoconvulsive response (PCR)results: GTCS in one case (contradictory to medical history),PGTCS in 11 cases (consistent with medical history),and GTCS and PGTCS hardly to distinguish in the remaining 3 cases. Of the three conditions above, there were generalized myoclonic seizures induced by IPS before TCS in 7 cases. CONCLUSION: The medical history was unreliable in determining whether TCS was generalized or focal. Myoclonic seizures can coexist with PGTCS, and sometimes GTCS was indistinguishable from PGTCS, indicating that the dichotomy of seizure types need to be improved. Photosensitive TCS should be regarded as a continuum between focal and generalized seizures.

[Application of scalp-recorded high-frequency oscillations in epileptic encephalopathy with continuous spike-and-wave during sleep].

OBJECTIVE: To investigate the clinical significance of high-frequency oscillations (HFOs) on scalp electroencephalography (EEG) in patients with epileptic encephalopathy with continuous spike-and-wave during sleep (CSWS). METHODS: Twenty-one CSWS patients treated for epilepsy from January 2006 to December 2016 in Pediatric Department of Peking University First Hospital were enrolled into the study. Selected clinical variables including gender, age parameters, seizure frequencies and antiepileptic drugs were compared between (a). HFO-positive group and HFO-negative group before methylprednisolone treatment and (b). excellent seizure outcome group and not-excellent seizure outcome group after methylprednisolone treatment. Interictal HFOs and spikes in pre- and post-methylprednisolone scalp EEG were measured and analyzed. RESULTS: Before methylprednisolone treatment, there were 12 of 21 (57%) CSWS patients had HFOs, with a mean value 43.17 per 60 s per patient. The 12 patients with HFOs tended to have more frequent epileptic negative myoclonus/atonic/myoclonus/atypical absences than those without HFOs in a month before methylprednisolone treatment. A total of 518 HFOs and 22 592 spikes were found in the pre-methylprednisolone EEG data of 21 patients, and 441 HFOs (86%) were associated with spikes. The highest amplitudes of HFOs were significantly positively correlated with that of spikes (r=0.279, P<0.001). Rates reduced by methylprednisolone treatment were statistically significant for both HFOs (P=0.002) and spikes (P=0.006). The percentage of reduction was 91% (473/518) and 39% (8 905/22 592) for spikes and HFOs, respectively. The percentage of spike and HFOs changes was respectively 100% decrease and 47% decrease in the excellent seizure outcome group, and they were 79% decrease and 18% increase in the not-excellent seizure outcome group. CONCLUSION: Prevalence of HFOs might reflect some aspect of epileptic activity. HFOs were more sensitive to methylprednisolone treatment than spikes and had a good correlation with the prognosis of seizures, and HFOs could be applied to assess epilepsy severity and antiepileptic therapy.

[Effects of monobutyl phthalate on migration and invasion of mouse Leydig tumor cells].

Objective: This study aimed to investigate the influence of monobutyl phthalate (MBP) on the expressions of epithelial-mesenchymal transition (EMT)-related proteins, migration and invasion of mouse Leydig tumor cells (MLTC-1) cells. Methods: After exposed to different doses of MBP (0、10(-7)、10(-6), 10(-5), 10(-4), 10(-3) mol/L) for 24 h or 48 h, cell viability was determined by 3-(4 5-dimethyl-2-thiazolyl)-2 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Expressions of vimentin, E-cadherin, N-cadherin and Snail proteins related to EMT were detected by Western blot. The ability of migration and invasion of MLTC-1 were assessed by wound healing assay and Transwell Boyden chamber assay, respectively. Results: Relative expressions of vimentin, Snail and N-cadherin proteins were promoted ((1.56±0.07) vs (1.78±0.08), (1.22±0.06) vs (1.44±0.07), (1.33±0.11) vs (2.19±0.06), all P values were<0.001) and E-cadherin (0.66±0.09) vs (0.47±0.06), P<0.001,protein was inhibited after the cells stimulated with MBP (0, 10(-7) and 10(-6) mol/L). The capability of wound closure of MLTC-1 cells were (6.64±2.07)%, (15.61±2.83)%, (39.91±0.33)%, respectively and the invading/migrating cells were (32.67±3.51), (57.67±2.52), (82.67±6.51), respectively, which were obviously increased under MBP treatments (0, 10(-7) and 10(-6) mol/L) (all P values were <0.001). Conclusion: Monobutyl phthalate affected the expressions of EMT-related proteins and enhanced the migration and invasion of MLTC-1 cells.

[The survival prediction model of advanced gallbladder cancer based on Bayesian network: a multi-institutional study].

Objective: To investigate the clinical value of Bayesian network in predicting survival of patients with advanced gallbladder cancer(GBC)who underwent curative intent surgery. Methods: The clinical data of patients with advanced GBC who underwent curative intent surgery in 9 institutions from January 2010 to December 2015 were analyzed retrospectively.A median survival time model based on a tree augmented naïve Bayes algorithm was established by Bayesia Lab software.The survival time, number of metastatic lymph nodes(NMLN), T stage, pathological grade, margin, jaundice, liver invasion, age, sex and tumor morphology were included in this model.Confusion matrix, the receiver operating characteristic curve and area under the curve were used to evaluate the accuracy of the model.A priori statistical analysis of these 10 variables and a posterior analysis(survival time as the target variable, the remaining factors as the attribute variables)was performed.The importance rankings of each variable was calculated with the polymorphic Birnbaum importance calculation based on the posterior analysis results.The survival probability forecast table was constructed based on the top 4 prognosis factors. The survival curve was drawn by the Kaplan-Meier method, and differences in survival curves were compared using the Log-rank test. Results: A total of 316 patients were enrolled, including 109 males and 207 females.The ratio of male to female was 1.0∶1.9, the age was (62.0±10.8)years.There was 298 cases(94.3%) R0 resection and 18 cases(5.7%) R1 resection.T staging: 287 cases(90.8%) T3 and 29 cases(9.2%) T4.The median survival time(MST) was 23.77 months, and the 1, 3, 5-year survival rates were 67.4%, 40.8%, 32.0%, respectively.For the Bayesian model, the number of correctly predicted cases was 121(≤23.77 months) and 115(>23.77 months) respectively, leading to a 74.86% accuracy of this model.The prior probability of survival time was 0.503 2(≤23.77 months) and 0.496 8(>23.77 months), the importance ranking showed that NMLN(0.366 6), margin(0.350 1), T stage(0.319 2) and pathological grade(0.258 9) were the top 4 prognosis factors influencing the postoperative MST.These four factors were taken as observation variables to get the probability of patients in different survival periods.Basing on these results, a survival prediction score system including NMLN, margin, T stage and pathological grade was designed, the median survival time(month) of 4-9 points were 66.8, 42.4, 26.0, 9.0, 7.5 and 2.3, respectively, there was a statistically significant difference in the different points(P<0.01). Conclusions: The survival prediction model of GBC based on Bayesian network has high accuracy.NMLN, margin, T staging and pathological grade are the top 4 risk factors affecting the survival of patients with advanced GBC who underwent curative resection.The survival prediction score system based on these four factors could be used to predict the survival and to guide the decision making of patients with advanced GBC.

[A multicenter retrospective study for the prognosis of T1b stage gallbladder carcinoma underwent different surgical procedure].

Objective: To explore the prognosis of patients with T1b stage gallbladder carcinoma underwent different surgical procedure. Methods: The clinicopathological data of 97 patients with T1b stage gallbladder carcinoma came from 8 clinical centers from January 2010 to December 2016 and 794 patients who were admitted to the SEER database of USA from January 1973 to December 2014 were analyzed.There were 891 patients including 254 males and 637 females (1.0∶2.5) with age of (69.5±12.0)years. There were 380 patients who were less than 70 years old, 511 patients who were more than 70 years old. And there were 213 patients with the diameter of tumor less than 20 mm, 270 patients with the diameter of tumor more than 20 mm, 408 patients were unclear. There were 196 patients with well differentiation, 407 patients with moderately differentiation, 173 patients with poorly differentiation, 8 patients with undifferentiated, 107 patients were unclear. In the 891 patients with T1b stage gallbladder carcinoma, there were 562 cases accepted the simple cholecystectomy, 231 cases with simple cholecystectomy plus lymphadenectomy, and 98 cases with radical cholecystectomy. The time of follow-up were until June 2017. χ(2) test was used to analyze the enumeration data, rank-sum test was used to analyze the measurement data, the analyses of prognostic factors were used Cox proportional hazards model, the survival analysis was performed using Kaplan-Meier method. Results: The results of Cox proportional hazards model indicated, age, differentiation, surgical procedure were the risk factors of prognostic(1.929(1.594-2.336), P<0.01; 1.842(1.404-2.416), P<0.01; 1.216(0.962-1.538), P<0.01). The results of Kaplan Meier test indicated, the overall survival of T1b stage gallbladder carcinoma were (85.5±3.8)months, the overall survival of patients with simple cholecystectomy were (71.3±4.4)months, the overall survival of patients with cholecystectomy plus lymphadenectomy were(87.6±5.8)months, and the overall survival of patients with radical cholecystectomy were(101.7±9.3)months. The overall survival of patients with cholecystectomy plus lymphadenectomy and radical cholecystectomy were more than simple cholecystectomy(P<0.05). There were 329 patients with Lymph nodes examined in and after operations(231 patients with cholecystectomy plus lymphadenectomy, 98 patients with radical cholecystectomy). There were 265 patients with negative lymph node metastasis, the overall survival were(98.3±4.2)months. There were 64 patients with positive lymph node metastasis, the overall survival were(75.5±3.1)months. The overall survival of 38 patients with cholecystectomy plus lymphadenectomy were(62.7±2.6) months, and 26 patients with radical cholecystectomy were (82.2±3.7)months. The overall survival of patients with radical cholecystectomy were more than cholecystectomy plus lymphadenectomy(P<0.05). Conclusions: The T1b stage gallbladder carcinoma patients with cholecystectomy plus lymphadenectomy or radical cholecystectomy has improved the prognosis comparing with simple cholecystectomy, significantly. When lymph node metastasis occurs, radical cholecystectomy has improved the prognosis comparing with cholecystectomy plus lymphadenectomy.

[Relationship between circulating tumor cells and coagulation factors in primary lung cancer patients].

OBJECTIVE: To investigate the relationship between the presence of circulating tumor cells (CTC) and the values of coagulation factors including D-dimer (D-D), fibrinogen (FIB) and platelet (PLT) in primary lung cancer patients. METHODS: Peripheral venous blood samples were collected from 79 patients with previously untreated primary lung cancer. The levels of D-D, FIB and PLT were detected. The CTCs were enriched by negative immunomagnetic separation with anti-CD45 antibody and then detected by immunocytochemistry with Anti-pan Cytokeratin antibody. The relationship between these parameters and clinicopathologic characteristics of the patients was analyzed. RESULTS: The levels of D-D, FIB and PLT were( 1.74±2.04) mg/L, (3.51±1.46 )g/L, (311±139)×10(9)/L, respectively. The level of D-D was associated with distant metastasis of lung cancer (P=0.046). The level of FIB was associated with clinical stage and distant metastasis (P<0.05). The level of PLT was associated with age, clinical stage and distant metastasis (all P<0.05). Among the 79 patients, there were 45 CTC-positive and 34 CTC-negative cases. The levels of D-D in the CTC-positive and CTC-negative groups were (2.31±2.41)mg/L and (0.99±1.02)mg/L, those of FIB were (3.79±1.56)g/L and (3.14±1.25)g/L, and those of PLT were (338±130)×10(9)/L and (229±129)×10(9)/L, respectively(P<0.05 for all). The positive rate of CTC was significantly higher in the metastasis group (82.8%), significantly higher than that in the non-metastatic group (42.0%, P<0.001). The levels of D-D, FIB and PLT in the metastasis group were (2.33±1.95)mg/L, (4.13±1.43)g/L and (433±74)×10(9)/L, but were (1.40±2.03) mg/L, (3.15±1.37)g/L and (206±88)×10(9)/L in the non-metastatic group (P<0.05 for all). CONCLUSIONS: The detection of circulating tumor cells may facilitate early prediction of distant metastasis of lung cancer. The hypercoagulation state is more-likely correlated with the distant metastasis of lung cancer.

[The effect of curcumin on the expression of miR-155 to apoptosis and invasion of extravillus trophoblast cells treated by lipopolysaccharide].

Objective: To investigate the effect of curcumin on the expression of miR-155, apoptosis and invasion of extravillus trophoblast cells treated by lipopolysaccharide (LPS). Methods: Human trophoblast cells (HTR-8/SVneo cells) were divided into 4 groups, the curcumin + LPS group (pre-treated by curcumin of 12.5, 25, 50 µmol/L, then LPS of 100 ng/ml), the LPS group (100 ng/ml), the recombinant adenovirus group (miR-155, multiplicity of infection100) and the control group. The miR-155 level in HTR-8/SVneo cells was measured by real-time PCR, and the expression of NF-κB was analyzed by luciferase gene expression. The apoptosis of HTR-8/SVneo cells was tested by cell death detection ELISA and the level of NF-κB in HTR-8/SVneo cells was measured by western blot. In addition, transwell was used to test the invasive ability of HTR-8/SVneo cells in all the groups. Results: (1) The intracellular expression of miR-155 in the LPS group was (2.13±0.22) times of the control group (P<0.01); and the expressions of miR-155 in 12.5, 25, 50 µmol/L curcumin+LPS groups were (0.37±0.08) , (0.68±0.14) , (0.49±0.09) times as the LPS group, with statistically significant difference (P<0.05). (2) The expreesion of NF-κB in the LPS group was (2.25±0.56) times of the control group. The expreesion of NF-κB in the 12.5, 25, 50 µmol/L curcumin+LPS groups were (0.80±0.07) , (0.74±0.05) , (0.49±0.19) times to the LPS group, with statistically significant difference(P<0.01). (3)The p65 protein in the LPS group was (1.50±0.22) , significantly higher than the control group (0.95±0.25, P<0.01) . In 12.5, 25, 50 µmol/L curcumin+LPS groups, the p65 protein were (0.31±0.07) , (0.75±0.14) , (0.49±0.08) . Compared with the LPS group, p65 was down-regulated by curcumin, with statistically significant difference (P<0.05). (4) In the cell death detection ELISA, the A value in the control group, the LPS group and the recombined adenovirus group were (0.89±0.17) , (2.02±0.35) and (1.67±0.48) , respectively, with statistically significant difference (P<0.05). In the curcumin+LPS groups, when the curcumin concentrations were 25 or 50 µmol/L, the A value were (0.74±0.05) , (0.49±0.09) , respectively, compared with the LPS group(set as 1.00), with statistically significant difference (P<0.05). When the curcumin concentration was 12.5 µmol/L, the A value was (0.80±0.07) , with no statistical significance (P>0.05). (5) The transmembrane cells were (47±8), (60±14) in the LPS group and the recombined adenovirus group, respectively. Compared with the control group (169±18), the differences were statistically significant (P<0.05). In the curcumin + LPS groups, the transmembrane cells were (143±10), (137±10) when the curcumin concentrations were 12.5, 25 µmol/L, significantly higher than the LPS group (P<0.05) . The transmembrane cells were (55±7) when the curcumin concentration was 50 µmol/L, with no statistical difference(P>0.05). Conclusion: The treatment of curcumin could downregulate the expression of NF-κB/miR-155, thus inhibit NF-κB signal pathway and the apoptosis of extravillus trophoblast cells, and protect their invasive ability.