Diagnosis and genetic analysis of a case with Bardet-Biedl syndrome caused by compound heterozygous mutations in the BBS12 gene.

Bardet-Biedl syndrome (BBS) is a rare autosomal recessive ciliopathy, which is caused by mutations mainly in genes encoding BBSome complex and IFT complex. Here, we reported a 21-year-old female with BBS characterized by three primary features including obesity, retinitis pigmentosa sine pigmento and bilateral renal cysts. She also had some secondary features such as diabetes mellitus, nonalcoholic fatty liver disease, subclinical hypothyroidism and mild conductive hearing damage. Whole exome sequencing revealed two compound heterozygous mutations in exon 2 of the BBS12 gene (c.188delC, p.T63fs and c.1993_1995del, p.665_665del) in this patient. Sanger sequencing showed that her father and mother carried c.188delC (p.T63fs) and c.1993_1995del (p.665_665del) variants, respectively, while her parents were free of BBS-related symptoms. In conclusion, this case reported two novel mutations (c.188delC, p.T63fs and c.1993_1995del, p.665_665del) of the BBS12 gene in a girl presented with BBS, which provides novel genetic resources for studies of the disease. Meanwhile, the BBS case shows the entire development progress from her birth to adulthood, which helps facilitate clinicians' understanding of BBS.

Traditional Chinese medicine formula Qing Huo Yi Hao as superoxide anion scavenger in high glucose-treated endothelial cells.

AIM: To investigate the effects of a traditional Chinese medicine formula Qing Huo Yi Hao (QHYH) and its components on hydroxyl radical (HO(•)) production in vitro and the activity of QHYH against free radicals in cultured endothelial cells induced by high glucose. METHODS: Hydroxyl radicals (HO(•)) were generated through Fenton reactions in vitro, and 5,5-dimethyl-1-pyrroline N-oxide (DMPO) was used as a spin trap to form DMPO/HO(•) adducts detected using electron paramagnetic resonance (EPR). Immortalized mouse cerebral microvascular endothelial (bEnd.3) cells were treated with high glucose (35 mmol/L). The free radical scavenging ability of QHYH in the cells was evaluated using EPR. Superoxide dismutase (SOD) was used to identify the free radicals scavenged by QHYH in the cells. RESULTS: QHYH and its 8 components concentration-dependently reduced DMPO/HO(•) signaling. The DMPO/HO(•) adduct scavenging ability of QHYH was 82.2%, which was higher than each individual component. The free radical scavenging ability of 1% QHYH in high glucose-treated bEnd.3 cells was approximately 70%. In these cells, the free radicals were also specifically reduced by SOD (400 U/mL), implying that the free radicals were primarily superoxide anions. CONCLUSION: The results demonstrate that the QHYH formula is potent antioxidant acting as scavenge of superoxide anions in high glucose-treated endothelial cells.

[Correlation of serum uric acid and islet beta cell functions in female type 2 diabetics].

OBJECTIVE: To examine the correlation of serum uric acid and islet beta cell functions in female type 2 diabetics. METHODS: A total of 533 female type 2 diabetics were recruited. And their clinicobiochemical parameters were measured. The levels of acute insulin response (AIR) and acute C-peptide response (ACPR) were measured by a stimulation of arginine. Among them, 262 patients received the examinations of OGTT (oral glucose tolerance test), area under the curve of insulin (AUC-insulin) and C peptide (AUC-C peptide). Serum uric acid > 360 µmol/L was defined as hyperuricemia. RESULTS: The serum uric acid levels of the hyperuricemia and control groups were (430 ± 8) and (248 ± 3) µmol/L respectively. The AIR, ACPR, AUC-insulin and AUC-C peptide levels were significantly higher in the hyperuricemia group than those in the control group (P < 0.01). There was a significant positive correlation between serum uric acid and AIR, ACPR, AUC-insulin and AUC-C peptide (r = 0.194, P < 0.01; r = 0.146, P < 0.01; r = 0.307, P < 0.01 and r = 0.420, P < 0.01). The serum levels of uric acid were significantly different between tertile groups of AIR (260 ± 7), (264 ± 8), (302 ± 7) µmol/L, ACPR ((263 ± 8), (271 ± 7), (296 ± 7) µmol/L), AUC-insulin ((241 ± 10), (279 ± 10), (301 ± 8) µmol/L) and AUC-C peptide ((229 ± 8), (265 ± 9), (326 ± 10) µmol/L, P < 0.01). Multivariate linear regression demonstrated that the serum level of uric acid were associated with AIR (ß = 0.002, 95%CI 0.0010 - 0.0030, P < 0.01), ACPR (ß = 0.001, 95%CI 0.0002 - 0.0020, P = 0.02), AUC-insulin (ß = 0.002, 95%CI 0.0004 - 0.0030, P < 0.01) and AUC-C peptide (ß = 0.003, 95%CI 0.0020 - 0.0030, P < 0.01) after an adjustment of related risk factors. CONCLUSION: The serum level of uric acid is significantly correlated with AIR, ACPR, AUC-insulin and AUC-C peptide in female type 2 diabetics. And it may be an independent risk factor of predicting islet beta cell functions in female type 2 diabetics.

Association of the aldosterone synthase gene -344T>C polymorphism with essential hypertension and glucose homeostasis: a case-control study in a Han Chinese population.

1. Aldosterone is a hormone that affects both blood pressure and glucose homeostasis. We studied the association of the aldosterone synthase gene (CYP11B2) polymorphism -344T>C with essential hypertension (EH) and glucose homeostasis in people in China. 2. We investigated the polymorphism -344T>C in CYP11B2 using a case-control study design (1059 cases and 1120 controls). Genotyping was carried out by matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy using a MassARRAY platform. 3. The aldosterone synthase gene -344T>C polymorphism was found to be associated with EH (odds ratio 1.252; 95% confidence interval 1.067-1.468; P(add) = 0.006). The -344C variant was found to be significantly associated with increased systolic blood pressure (P(add) = 0.003) and diastolic blood pressure (P(add) = 0.024) in controls and increased plasma aldosterone levels (P(add) = 0.0001) in EH cases. The -344C variant was also found to be significantly associated with increased fasting glucose (P(rec) = 0.003) in controls. In the subgroup containing 893 EH cases without a history of diabetes or hypoglycaemia medications, the -344C variant was found to be associated with increased fasting and postprandial plasma glucose levels (P(add) = 0.0001, P(add) = 0.001, respectively) and decreased pancreatic ß-cell function and insulin sensitivity (P(add) = 0.030, P(dom) = 0.019, respectively) by homeostasis model assessments. 4. In this Han Chinese population, the -344T/C polymorphism of the CYP11B2 gene was found to be associated with EH and glucose homeostasis, both of which might be mediated by plasma aldosterone levels, insulin sensitivity and ß-cell function.

Hypoxia-Induced Glioma-Derived Exosomal miRNA-199a-3p Promotes Ischemic Injury of Peritumoral Neurons by Inhibiting the mTOR Pathway.

The underlying molecular mechanisms that the hypoxic microenvironment could aggravate neuronal injury are still not clear. In this study, we hypothesized that the exosomes, exosomal miRNAs, and the mTOR signaling pathway might be involved in hypoxic peritumoral neuronal injury in glioma. Multimodal radiological images, HE, and HIF-1α staining of high-grade glioma (HGG) samples revealed that the peritumoral hypoxic area overlapped with the cytotoxic edema region and directly contacted with normal neurons. In either direct or indirect coculture system, hypoxia could promote normal mouse hippocampal neuronal cell (HT22) injury, and the growth of HT22 cells was suppressed by C6 glioma cells under hypoxic condition. For administrating hypoxia-induced glioma-derived exosomes (HIGDE) that could aggravate oxygen-glucose deprivation (OGD)/reperfusion neuronal injury, we identified that exosomes may be the communication medium between glioma cells and peritumoral neurons, and we furtherly found that exosomal miR-199a-3p mediated the OGD/reperfusion neuronal injury process by suppressing the mTOR signaling pathway. Moreover, the upregulation of miRNA-199a-3p in exosomes from glioma cells was induced by hypoxia-related HIF-1α activation. To sum up, hypoxia-induced glioma-derived exosomal miRNA-199a-3p can be upregulated by the activation of HIF-1α and is able to increase the ischemic injury of peritumoral neurons by inhibiting the mTOR pathway.

[Analysis of CCM1 gene mutations in Chinese patients with intracranial cavernous malformations].

OBJECTIVE: To study the CCM1 gene (7q11.2 - q22) mutations in Chinese patients with intracranial cavernous malformations (ICM), METHODS: Peripheral blood samples were collected from 25 unrelated patients with ICM confirmed by post-operational pathology, 7 being with familial ICM, all of Han nationality, and from 30 healthy people as controls. The genomic DNA was extracted and the exons 8, 9, 11, 12, 13, 15, 16, 17, and 18 of CCM1 gene and part of intervening sequences near both sides of these exons were amplified by PCR. The PCR products were sequenced directly and then compared with the GenBank data. RESULTS: Seven new mutation sites of CCM1 gene were detected from 11 Chinese ICM patients with a total mutation rate of 44%. Of the seven new mutations there were three missense mutations: 1160A-->C (Q387P) and 1172C-->T (S391F) in exon12, and 1405A-->C (N469H) in exon13; two insertion mutations: 704insT (K246stop) in exon8, and 2138insG (T733stop) in exon18; one intervening sequence mutation: IVS12 - 4C-->T; and one synonymous mutation: 1875C-->T (F625F) in exon17. None mutation was detected in the control group. The CCM1 mutation rate of familial ICM was 85.7%, significantly higher than that of sporadic ICM (27.7%, P < 0.05). CONCLUSION: As the genetic basis of ICM, CCM1 gene mutation exists in Chinese ICM patients too, that leads to functional loss or changes of the gene encoding KRIT1 protein.

Cross-sectional association of serum C-reactive protein and uric acid with albuminuria in Chinese type 2 diabetic patients.

BACKGROUND: Evidences show that subclinical chronic inflammation is involved in the pathogenesis of diabetic nephropathy. The aim of this study was to examine the relationship between serum C-reactive protein (CRP), serum uric acid, and albuminuria in Chinese type 2 diabetic patients. METHODS: A total of 1162 type 2 diabetic patients were recruited. All participants had relevant clinical and laboratory measurements. CRP was measured using a particle enhanced immunoturbidimetric assay. RESULTS: In the multiple linear regression model, natural log-transformed CRP (lnCRP) and uric acid were independent predictors of natural log-transformed urinary albumin to creatinine ratio (lnACR) (ß = 0.18, 95% CI 0.10-0.27, P < 0.001 and ß = 0.18, 95% CI 0.09-0.27, P < 0.001). The interaction of lnCRP with uric acid was also associated with lnACR (ß = 0.04, 95% CI 0.02-0.06, P < 0.001). In the full-adjusted logistic regression model, the OR for albuminuria of the patients in the third tertile levels of CRP and uric acid was 3.94 compared with patients in the first tertile levels of CRP and uric acid (95% CI 1.73-8.94, P < 0.001). CONCLUSIONS: Elevated serum CRP and increased serum uric acid level were associated with albuminuria in Chinese type 2 diabetic patients. Moreover, CRP and uric acid had an interactive effect on albuminuria.

The obesity-related polymorphism PCSK1 rs6235 is associated with essential hypertension in the Han Chinese population.

Proprotein convertase subtilisin/kexin-type 1 (PCSK1) is a prohormone convertase that has an important role in prohormone maturation including the process of prorenin to renin. We studied the association of the PCSK1 single-nucleotide polymorphism (SNP) rs6235 (encoding an S690T substitution) with essential hypertension (EH), obesity and related traits in the Han Chinese population. The rs6235 SNP in the PCSK1 gene was investigated using a case-control study design, with 1034 hypertension cases and 1112 normotensive controls. In this study, the rs6235 SNP was significantly associated with hypertension (OR=1.26, 95% CI (1.10-1.46), P=0.001); the odds ratios of GC vs GG and CC vs GG were 1.30 (95% CI (1.06-1.58), P=0.010) and 1.55 (95% CI (1.12-2.13), P=0.007), respectively. In the controls, the C-allele was associated with increased systolic (P=0.010) and diastolic (P=0.010) blood pressure levels. In all of the EH patients and EH patients without a history of renin-angiotensin-aldosterone (RAA) system-related antagonists, the C-allele was associated with increased plasma renin activity (P=0.00004 and 0.002, respectively) and aldosterone levels (P=0.018 and 0.005, respectively). The C-allele was also associated with increased body mass index (BMI) (P=0.010) in the normotensive controls. In conclusion, the PCSK1 SNP rs6235 was associated with EH and blood pressure in the Han Chinese population, and this association may be mediated by the SNP's effect on RAA levels. rs6235 was also associated with BMI in this population.

Tetramethylpyrazine protects palmitate-induced oxidative damage and mitochondrial dysfunction in C2C12 myotubes.

AIMS: Tetramethylpyrazine (TMP), one of the active ingredients isolated from a Chinese herbal prescription, possesses protective effects against oxidative stress caused by high glucose in endothelial cells. In this study, the role of TMP in preventing muscle cells from palmitate-induced oxidative damage was investigated and the possible mechanisms of action elucidated. MAIN METHODS: Mitochondrial reactive oxygen species (ROS) were measured in C2C12 myotubes, a palmitate-induced oxidative stress cell model, with or without TMP. Both mitochondrial membrane potential (MMP) and oxygen consumption were assessed in conjunction with quantification of mitochondrial DNA and mitochondrial biogenesis-related factors, such as peroxisome proliferator-activated receptor-γ coactivator 1 α (PGC1α), nuclear respiratory factor 1 (NRF1) and mitochondrial transcription factor A (Tfam), by real-time polymerase chain reaction. Expression of mitochondrial respiratory chain complex III as an index of mitochondrial function was evaluated by immunoblotting, and glucose transport into the C2C12 myotube examined by analyzing 2-deoxy-[(3)H]glucose uptake. KEY FINDINGS: TMP significantly alleviated palmitate-induced mitochondrial ROS production, mitigated mitochondrial dysfunction and increased D-loop mRNA expression as compared with the control. This was accompanied by a marked reversal of palmitate-induced down-regulation in the expression of mitochondrial biogenesis-related factors (PGC1α, NRF1 and Tfam) and decreased glucose uptake in C2C12 myotubes. As a result, cell respiration, as reflected by the elevated expression of mitochondrial respiratory chain complex III and oxygen consumption, was enhanced. SIGNIFICANCE: TMP is capable of protecting C2C12 myotubes against palmitate-induced oxidative damage and mitochondrial dysfunction, and improving glucose uptake in muscle cells partially through the up-regulation of mitochondrial biogenesis.