Analysis of gut microbiota in patients with epilepsy treated with valproate: Results from a three months observational prospective cohort study.

BACKGROUND: Growing evidence implicates the potential effect of microbiota on the pathogenesis and course of epilepsy. However, the effects of valproate (VPA), a broad spectrum anti-epileptic drugs, on gut microbiota have not been investigated in humans. This study aimed to analyze fecal microbiota in patients with epilepsy treated with valproate. METHODS: A total of 10 participants, who were newly diagnosed of cryptogenic epilepsy with treatment naïve and received 1000 mg daily doses of VPA, were recruited in our prospective study. Microbiota compositions were evaluated at baseline and after three months of VPA treatment using 16S rDNA sequencing. RESULTS: VPA treatment was associated with clinical improvements in all patients, but not changes in gut microbiota richness and complexity (Shannon: p = 0.82). Microbiome composition structure differences also revealed no statistical difference in dissimilarity (Adonis: p = 0.90). No statistical difference taxa were found between two groups. However, the ratio of phyla Firmicutes to Bacteriodetes (ANOVA: p = 0.037) markedly raised after three months of VPA-treatment. A correlation matrix based on the spearman correlation distance confirmed associations between specific fecal taxa and VPA-related clinical metabolic parameters, including drug concentration in the blood, total cholesterol, triglyceride, lactate dehydrogenase, alanine aminotransferase, aspartate aminotransferase and weight gain. (p < 0.05) CONCLUSIONS: Among those patients treated with VPA, characterization of the gut microbiota altered, and gut microbiota associated with weight gain and clinical biochemical indexes, suggesting that microbiome composition data might involve in the mechanisms of VPA induced metabolic disorder.

HLA-DRB1*03:01 is associated with female sex and younger age of anti-LGI1 encephalitis.

BACKGROUND AND PURPOSE: This study was undertaken to investigate the association between human leukocyte antigen (HLA) genotype and clinical characteristics of anti-LGI1 encephalitis. METHODS: HLA genotyping was performed in 34 patients with anti-LGI1 encephalitis admitted to West China Hospital between April 2014 and May 2021, as well as in 305 healthy controls. The online tool NetMHCIIpan 4.0 and AutoDock Vina software were used to predict binding between LGI1 peptide and HLA class II molecules. RESULTS: Risk of anti-LGI1 encephalitis was strongly associated with the DRB1*03:01 allele (odds ratio [OR] = 4.31, 95% confidence interval [CI] = 1.96-9.25, corrected p = 2.75 × 10-4 ) and the DRB1*03:01-DQB1*02:01 haplotype (OR = 4.45, 95% CI = 2.02-9.59, corrected p = 2.94 × 10-4 ). Compared to carriers of the DRB1*07:01 allele, those with the DRB1*03:01 allele were more likely to be female (93.3% vs. 33.3%, p = 0.004) and to be younger (median age = 38 vs. 65 years, p < 0.001). DRB1*03:01 carriers showed stronger response to immunotherapy than carriers of the DRB1*07:01 allele, based on median score decrease on the modified Rankin scale (2, interquartile range = 1-2 vs. 1, interquartile range = 0-1; p = 0.03) at 4 weeks after immunotherapy. Prediction and docking algorithms suggested that the LGI1 peptide can bind to the DRB1*03:01 molecule strongly. CONCLUSIONS: The strong association between anti-LGI1 encephalitis and certain HLA class II alleles supports a primary autoimmune origin for the disease. Carriers of the DRB1*03:01 allele in Chinese patients with anti-LGI1 encephalitis are more likely to be female, to suffer earlier disease onset, and to respond better to immunotherapy.

The neuroprotective effect of apigenin against OGD/R injury in rat hippocampal neurons.

To investigate the effects of apigenin on the injury caused by oxygen and glucose deprivation in neurons and the underlying mechanisms, primary cultured rat hippocampal neurons were incubated with apigenin for 90 min before a 2-h oxygen and glucose deprivation followed by a 24-h reperfusion (OGD/R). Subsequently, cell viability, lactate dehydrogenase (LDH) leakage rate, apoptotic rate of neurons and activity of the sodium pump were assessed. In addition, activity of the sodium pump was also examined in the hippocampus of SD rats injected intraperitoneally with apigenin 90 min before a 10-min global cerebral ischemia/24-h reperfusion. The results showed that cell viability and activity of the sodium pump markedly decreased but LDH leakage rate and apoptotic rate significantly increased in OGD/R-treated neurons. However, pretreatment with apigenin (20-50µmol/L) reversed the changes dose-dependently. Compared to sham controls, activity of the sodium pump was significantly suppressed in global ischemia/reperfusion rats; application of apigenin (200mg/kg) restored the activity of the sodium pump. Furthermore, the neuroprotective effect of apigenin was blocked partly by the sodium pump inhibitor ouabain. Our findings provide the evidence that apigenin has a neuroprotective effect against OGD/R injury and the protective effect may be associated with its ability to improve sodium pump activity.

[Construction of COL1A1 short hairpin RNA vector and its effect on cell proliferation and migration of gastric cancer cells].

OBJECTIVE: To construct COL1A1-targeted short hairpin RNA (shRNA) vector with pSilencer 4.1-CMV neo siRNA expression vector and to evaluate its effect on proliferation and migration of gastric cancer BGC-823 cells in vitro. METHODS: Three COL1A1-shRNA plasmids (COL1A1-shRNA-1, COL1A1-shRNA-2, COL1A1-shRNA-3), targeting different sites of COL1A1 gene, were constructed using pSilencer 4.1-CMV neo siRNA expression vector and transfected into gastric cancer BGC-823 cells. Real time quantitative RT-PCR and Western blot were performed to detect expression levels of COL1A1. MTT and Transwell migration assays were employed to evaluate the effects of COL1A1 gene silence on cell proliferation and migration. RESULT: Three recombinant plasmids targeting COL1A1 were constructed successfully. The expressions of COL1A1 in BGC-823 cells, including mRNA and protein levels, were significantly inhibited by the COL1A1-shRNA transfectants, which resulted in a clear reduction of cell proliferation and migration capacity. CONCLUSION: The COL1A1-shRNA can effectively knock down gene expression and inhibit proliferation and migration of gastric cancer BGC-823 cells.

Collagen: a possible prediction mark for gastric cancer.

Considering the high mortality rates and the unfavorable prognosis of gastric cancer (GC), it is important to predict early GC canceration and the metastasis. Unfortunately, it has not been any clinical prediction mark sufficiently sensitive to GC yet. It is therefore important to investigate new sensitive and specific marks for GC prediction. Here, we review the evidence on gastric cancer and collagen, and discuss the sensitivity and the feasibility of collagens as potential prediction marks. Experiment researches and micro-array technology have shown that not only the biosynthesis and degradation of collagens but also collagen genes' expression are closely related to GC biological behaviour, especially canceration and metastasis and studies have proved some certain collagens might have ability to predict GC development. As outlined above, we hypothesize that collagens may be independent prediction marks of GC canceration and metastasis.

Screening for metronidazole-resistance associated gene fragments of H pylori by suppression subtractive hybridization.

AIM: To screen for metronidazole (MTZ)-resistance associated gene fragments of H pylori by suppression subtractive hybridization (SSH). METHODS: Five MTZ-resistant (tester, T) and 1 MTZ-susceptible (driver, D) clinical H pylori isolates were selected. Genomic DNAs were prepared and submitted to Rsa I digestion. Then two different adaptors were ligated respectively to the 5'-end of two aliquots of the tester DNA fragments and SSH was made between the tester and driver DNAs. The specific inserts of tester strains were screened and MTZ-resistance related gene fragments were identified by dot blotting. RESULTS: Among the randomly selected 120 subtractive colonies, 37 DNA fragments had a different number of DNA copies (> or = 2 times) in resistant and susceptible strains and 17 of them had a significantly different number of DNA copies (> or = 3 times). Among the sequences obtained from the 17 DNA fragments, new sequences were found in 10 DNA fragments and duplicated sequences in 7 DNA fragments, representing respectively the sequences of depeptide ABC transporter periplasmic dipeptide-binding protein (dppA), permease protein (dppB), ribosomal protein S4 (rps4), ribonuclease III (rnc), protease (pqqE), diaminopimelate epimerase (dapF), acetatekinase (ackA), H pylori plasmid pHP51 and H pylori gene 1334. CONCLUSION: Gene fragments specific to MTZ-resistant H pylori strains can be screened by SSH and may be associated with MTZ-resistant H pylori.

[Screening for metronidazole-resistance associated gene fragments of Helicobacter pylori by suppression subtractive hybridization].

OBJECTIVE: To screen metronidazole (MTZ) -resistance associated gene fragments of Helicobacter pylori (H.pylori) by suppression subtractive hybridization(SSH). METHODS: The suppression subtractive hybridization (SSH) was used to screen the different DNA fragments between MTZ-resistant and -susceptible clinical strains of H.pylori. The resistant strains specific gene fragments were obtained by SSH and identified by dot blotting. RESULT: Among the 120 subtractive colonies which were randomly chosen, 37 DNA fragments were different (>or=2 times) in DNA-copy numbers between resistant and susceptible strains and 17 of them were significantly different (>or=3 times). These 17 DNA fragments were sequenced subsequently. Ten of them were new sequences and the other 7 were duplicated sequences. These sequences represented respectively: depeptide ABC transporter periplasmic dipeptide-binding protein (dppA), permease protein (dppB), et al. CONCLUSION: Gene fragments specific to MTZ-resistant H. pylori strains were obtained by SSH and these genes may associated with MTZ-resistance of H.pylori.

How reliable is the Ki-67 cytological index in grading pancreatic neuroendocrine tumors? A meta-analysis.

OBJECTIVE: To investigate the accuracy of the cytological Ki-67 index in distinguishing intermediate and high-grade (G2 + G3) from low-grade (G1) pancreatic neuroendocrine tumors (PNETs). METHODS: Two investigators independently searched databases to identify eligible studies using the following term: ('Ki-67') AND ('pancreatic endocrine tumor' OR 'pancreatic neuroendocrine tumor' OR 'pancreatic endocrine tumour' OR 'pancreatic neuroendocrine tumour' OR 'pancreatic endocrine tumors' OR 'pancreatic neuroendocrine tumors' OR 'pancreatic endocrine tumours' OR 'pancreatic neuroendocrine tumours'), and meta-analysis was performed to calculate the pooled sensitivity, specificity, positive (PLR) and negative likelihood ratio (NLR), and diagnostic odds ratio (DOR). RESULTS: A total of 263 lesions from 13 studies were included in the study. The pooled sensitivity and specificity of Ki-67 (cut-off value: 2%) in the differential diagnosis of G2 + G3 from G1 PNETs were 64% and 87%, respectively. The pooled PLR, NLR and DOR were 3.96, 0.42 and 11.21, respectively. The area under the summary receiver operating characteristic curve (AUROC) was 0.8397. While the cut-off value of Ki-67 index was set as 5%, the sensitivity and specificity were increased up to 69% and 93%, respectively, and the AUROC was increased to 0.955. CONCLUSION: The cytological Ki-67 index is very useful in distinguishing intermediate and high-grade from low-grade PNETs, and a cut-off value of 5% had a better predictive value compared with that of 2%.

Overweight influence on circadian variations of ambulatory blood pressure in Chinese adolescents.

Childhood obesity and its consequences have been the subject of intense interest in recent years. In this study we examined the influence of overweight on circadian variations of ambulatory blood pressure (ABP) in Chinese adolescents. First, 24-hr ABP monitoring was performed in 252 adolescents divided into two groups with equivalent sex, age, and body height (49 girls and 77 boys in each group): controls (normal weight) were aged 13.68 +/- 1.21 years, height 165.37 +/- 9.45 cm, body mass index (BMI) 18.82 +/- 2.3; overweights (BMI > or = 24) were aged 13.71 +/- 1.23 years, height 165.75 +/- 9.47 cm, BMI 27.70 +/- 3.1. ABP recordings were treated by ABP database system and analyzed by cosinor method and conventional statistics methods. The circadian variations of ABP in adolescent patterned as "dipper" and circadian rhythmicity of ABP variations were confirmed by cosinor analysis in most adolescents of both groups. Significant statistical differences were found for rhythm parameters: the MESOR (midline estimate statistic of rhythm), peak, trough (the maximum and minimum values derived from the composed curves, respectively), and amplitude values between control and overweight groups. Significant higher values also were seen in the overweight group for most of ABP parameters (p < .01), such as, BP means (SBP, DBP, MAP: mean arterial pressure, or PP: pulse pressure), BP variability, BP loads and rate-pressure product (HR x SBP). Our results have shown that overweight influenced significantly on ABP and parameters derived from ABP recordings in Chinese adolescents, which suggests an increasing risk of cardiovascular diseases in overweight adolescents.