[Genotype-phenotype analysis of Fabry disease caused by GLA gene variation in a pedigree].

Objective: To investigate the clinical phenotype and genetic characteristics of patients with Fabry disease caused by a GLA variant, IVS4+919G>A. Methods: It was a prospective study. Fabry disease screening was conducted among high-risk population in Ninghai from October 2021 to August 2023. Those children with decreased α-galactosidase enzyme activity<2.40 µmol/(L·h) or elavated Lyso-GL-3 level>1.10 µg/L in dried blood spot (DBS) method underwent GLA genetic testing for diagnosis confirmation. Meanwhile, family screening was carried out. A proband and his family members diagnosed with Fabry disease were research subjects. The clinical and genetic characteristics of patients with Fabry disease caused by the GLA variant (IVS4+919G>A) were analyzed. Results: The female proband aged 9.8 years with pain in both lower limbs as the initial symptom was found to have a heterozygous GLA variant IVS4+919G>A among 102 patients. In family screening, there were 4 family members (proband's father, elder sister, elder male cousin and elder female cousin) with Fabry disease and a family member (proband's fifth aunt) with a GLA variant. Among these 4 diagnosed family members, the elder male cousin of the proband, a boy aged 13.2 years had a heterozygous GLA variant, IVS4+919G>A with intermittent pain in both lower limbs as the initial symptom. The proband's father had knee joint pain. The proband's elder sister had decreased vision and his elder female cousin had no obvious symptoms. The proband's fifth aunt with a GLA variant had decreased vision. Conclusions: High-risk screening in children and family screening are helpful for early diagnosis and treatment of Fabry disease. Neuropathic pain may be a early symptom in children with Fabry disease caused by the GLA variant, IVS4+919G>A.

Promethazine inhibits neuronal apoptosis via PI3K/Akt signaling pathway in rats with cerebral infarction.

OBJECTIVE: To study the effect of promethazine on neuronal apoptosis in rats with cerebral infarction (CI) through the phosphatidylinositol 3-hydroxy kinase/protein kinase B (PI3K/Akt) signaling pathway. MATERIALS AND METHODS: A total of 36 Sprague-Dawley rats were randomly divided into the sham group (n=12), model group (n=12), and promethazine group (n=12). The external carotid artery was only exposed in the model group, and the ischemia-reperfusion model after CI was established using the suture method in the other two groups. After modeling, the normal saline was intraperitoneally injected in the sham group and model group, while promethazine was intraperitoneally injected in the promethazine group. The rats were sampled after 1 week of intervention. The neurological deficits of rats were evaluated using the Zea-Longa score, and the cognitive function, the spatial learning, and memory of rats were detected via the water maze test. Moreover, the expressions of B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) in brain tissues were detected via immunohistochemistry, and the relative protein expressions of PI3K p85, PI3K p110, and p-Akt were detected via Western blotting. The mRNA expressions of Bax and Bcl-2 were detected via quantitative Polymerase Chain Reaction (qPCR), and the apoptosis was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. RESULTS: The Zea-Longa score was significantly increased in the model group and promethazine group compared with that in the sham group (p<0.05), while it significantly declined in the promethazine group compared with that in the model group (p<0.05). The escape latency was significantly prolonged and the times of crossing platform were significantly reduced in the model group and promethazine group compared with those in the sham group (p<0.05), while the escape latency was significantly shortened and the times of crossing platform were significantly increased in the promethazine group compared with those in the model group (p<0.05). Compared with those in the sham group, the positive expression of Bax was significantly increased, while the positive expression of Bcl-2 was remarkably decreased in the model group and promethazine group (p<0.05). Compared with those in the model group, the positive expression of Bax was significantly decreased, while the positive expression of Bcl-2 was remarkably increased in the promethazine group (p<0.05). Besides, the model group and promethazine group had evidently higher relative protein expressions of PI3K p85, PI3K p110, and p-Akt than the sham group (p<0.05), while the promethazine group also had evidently higher relative protein expressions of PI3K p85, PI3K p110, and p-Akt than the model group (p<0.05). Compared with the sham group, model group, and promethazine group had remarkably increased relative mRNA expression of Bax, and remarkably decreased relative mRNA expression of Bcl-2 (p<0.05). Compared with those in the model group, the relative mRNA expression of Bax was remarkably decreased, while the relative mRNA expression of Bcl-2 was remarkably increased in the promethazine group (p<0.05). Finally, the apoptosis rate was significantly higher in the model group and promethazine group than that in the sham group (p<0.05), while it was significantly lower in the promethazine group than that in the model group (p<0.05). CONCLUSIONS: Promethazine inhibits neuronal apoptosis in CI rats by upregulating the PI3K/Akt signaling pathway, thereby exerting a protective effect.

Metformin regulates tight junction of intestinal epithelial cells via MLCK-MLC signaling pathway.

OBJECTIVE: To observe the effect of metformin on the tight junction of intestinal epithelial cells and its relevant mechanism. MATERIALS AND METHODS: Caco-2 cell monolayers were incubated with or without tumor necrosis factor-α (TNF-α) (10 ng/mL) in the absence or presence of indicated concentrations of metformin. Transepithelial electrical resistance (TEER) was measured at various time points. Caco-2 cell permeability was assessed using fluorescein permeability test. Immunofluorescence was used to detect the distribution of tight junction protein. Western blotting and Real-Time PCR were used to detect the expression of tight junction protein and Myosin light chain kinase (MLCK)-Myosin light chain (MLC) signaling pathway. RESULTS: Metformin attenuates the effects of TNF-α on Caco-2 cell TEER and paracellular permeability, prevents TNF-α-induced morphological disruption of tight junctions, ameliorates the inhibiting effect of TNF-α on epithelial tight junction-related protein expression and suppresses the TNF-α-induced increase in MLCK production. CONCLUSIONS: Metformin can stabilize and up-regulate tight junction protein by inhibiting MLCK-MLC signaling pathway, thus ameliorating the tight junction of intestinal epithelial cells.

[Application of Chinese version of ACE-â ¢ in type 2 diabetes mellitus patients with mild cognitive impairment].

Objective: To investigate the application and best cut-off value of Chinese version of Addenbrooke's cognitive examination-Ⅲ(ACE-Ⅲ) in type 2 diabetes mellitus (T2DM) patients with mild cognitive impairment. Methods: A total of 18 T2DM patients with normal cognitive function (NCI group) and 40 T2DM patients with mild cognitive impairment (MCI group) treated in outpatient clinic or ward of Department of Neurology and Endocrinology in Fujian Medical University Union Hospital between January 2015 and February 2016 were enrolled. Mini Mental State Scale (MMSE), Montreal cognitive assessment scale (MoCA), Activity of Daily Living Scale (ADL) and the Chinese version of ACE-Ⅲ were used to assess cognitive function of subjects and to assess the value of ACE-Ⅲ in the diagnosis of T2DM patients with mild cognitive impairment. Results: The Cronbach's alpha of the Chinese version of ACE-Ⅲ is 0.768. ACE-Ⅲ and MoCA were correlative (r=0.768, P<0.001). The area under the receiver operating characteristic (ROC) curve for ACE-Ⅲ was 0.906 (95%CI: 0.827-0.985). When the cut-off value for diagnosis was 87.5, the maximum Youden index was 0.769, with a sensitivity of 0.825 and a specificity of 0.944. Patients in MCI group got a lower score in the sub-items of attention/orientation, memory, verbal fluency, language and visual space of ACE-Ⅲ compared to those in NCI group, and the differences were statistically significant (t=5.336, P<0.001; t=5.530, P<0.001; t=4.556, P<0.001; t=5.301, P<0.001; t=2.821, P=0.008). Conclusion: The Chinese version of ACE-Ⅲ had good internal consistency reliability, and it could effectively detect impairment of general cognitive function and single cognitive domains in T2DM patients.

[Reoperation for residual aneurysm of coronary anastomosis after Bentall procedure].

Objective: To introduce a new operative method for residual aneurysm of coronary anastomosis after Bentall procedure. Methods: Between March 2011 and December 2012, six patients in Beijing Anzhen Hospital with residual aneurysm of coronary anastomosis (CT showed goldfish eye sign at the openings of coronary) after Bentall procedure underwent the operation of concentric circular patch procedure under cardiopulmonary bypass. Femoral artery, right atrium and upper right pulmonary artery cannulation were used for cardiopulmonary bypass, and the artificial vessel was transected after cardiac arrest. A concentric circular patch was pruned, whose outside diameter was slightly larger than the aneurysm and the inside diameter was equal to the openings of coronary. The outer edge of the patch was anastomosed to the outer edge of the aneurysm (opening of artificial vessel in primary surgery) with 4-0 prolene. The inner edge of the patch was anastomosed to the openings of coronary with 5-0 prolene. Results: All patients had clinical recovery. Postoperative CT demonstrated the disappearance of residual aneurysm during follow-up (the goldfish eye sign disappeared). Conclusion: The concentric circular patch procedure is a feasible treatment for residual aneurysm of coronary anastomosis.

[The relationship between preoperative renal failure and severe postoperative hypoxemia of patients received surgical procedures for Stanford A aortic dissection].

OBJECTIVE: To study the relationship between renal failure and severe postoperative hypoxemia of patients received surgical procedure for Stanford A aortic dissection. METHODS: Clinical data of 411 consecutive patients from January 2014 to April 2015, who received surgical procedure for Stanford A aortic dissection in Department of Cardiovascular Surgery of Beijing Anzhen Hospital, were collected retrospectively. The appearance of severe postoperative hypoxemia was recorded in all the cases. All the data about potential prognostic factors was put into the database and analyzed by univariate and multivariate Logistic regression respectively. RESULTS: Severe postoperative hypoxemia (PO2/FiO2<100 mmHg, 1 mmHg=0.133 kPa) happened on 69 cases within 48 hours after procedures, with the incidence rate of 17.1%. Both univariate and multivariate Logistic regression indicated the influence that preoperative creatinine clearance rate had on severe postoperative hypoxemia showed no statistical significance. However, the influence of preoperative serum creatinine showed statistical significance (OR=1.009, 95%CI: 1.000 to 1.018, P=0.048). CONCLUSIONS: The preoperative creatinine clearance rate of patients has no direct relationship with severe postoperative hypoxemia. But the preoperative serum creatinine could be regarded as an independent predictor of severe postoperative hypoxemia.

[Application of evoked potentials monitoring in total thoracoabdominal aorta aneurysm repair].

OBJECTIVE: To evaluate the application value of evoked potentials (EP) monitoring in patients undergoing aorta-iliac bypass for total thoracoabdominal aorta aneurysm repair (tTAAAR). METHODS: A prospective study, with a total of 31 patients undergoing tTAAAR and intraoperative EP monitoring from June 2014 to April 2015 was carried out. The results of intraoperative evoked potentials, clinical outcomes and follow-up data of patients were collected for further evaluation. RESULTS: The EP wave disappeared [motor evoked potentials for (55.6±18.1) min, somatosensory evoked potentials for (50.3±18.7) min] after proximal descending aorta being clamped, and gradually recovered after the segment arteries of spine cord were reconstructed. The EP wave was restored to normal level at the end of operation in all the cases. The somatosensory evoked potentials remained unchanged in 2 cases (false negative). One case died after operation. No spinal cord injury occurred. The median follow-up after operation was 10 months (5-14 months). There was no delayed neurological deficit. CONCLUSION: EP provided an on-line monitoring of the condition of spinal cord function, which become an intraoperative protocol to avoid the irreversible injury of spinal cord.

[A prediction model for severe postoperative hypoxemia after surgery for Standford type A aortic dissection].

OBJECTIVE: To study the risk factors of severe postoperative hypoxemia after surgery for Standford type A aortic dissection and establish a prediction model. METHODS: Data of 411 consecutive patients from January 2014 to April 2015, who underwent surgery for Standford type A aortic dissection in the department of cardiovascular surgery of Beijing Anzhen Hospital, were retrospectively analyzed. All the cases were divided into two groups according to the appearance of severe postoperative hypoxemia. All the data about potential risk factors was put into the database and analyzed by logistic regression. The prediction model was then established upon acquired independent risk factors. Discrimination and calibration of the prediction model were assessed with ROC curve and Hosmer and Lemeshow goodness of fit test. RESULTS: The perioperative in-hospital mortality was 6.57%(27/411). Severe postoperative hypoxemia (PaO2/FiO2≤100 mmHg) happened in 69 cases within 48 hours after procedures, with an incidence rate of 17.1%. The logistic regression demonstrated that body mass index (BMI), age, preoperative serum myoglobin, preoperative serum creatinine, preoperative serumalanine aminotransferase, the time of cardiopulmonary bypass, re-exploration within 48 hours after procedures were the independent risk factors for severe postoperative hypoxemia. The prediction model was then established based on these independent risk factors. The area under ROC curve of the model was 0.785, and the P value in Hosmer and Lemeshow goodness of fit test was 0.625. CONCLUSION: The logsitic model built in this study succeeded to predict the incidence of severe postoperative hypoxemia after surgery for Standford type A aortic dissection, and it could meet the doctors' requirement with its excellent discrimination and calibration.

Increased vesicular γ-GABA transporter and decreased phosphorylation of synapsin I in the rostral preoptic area is associated with decreased gonadotrophin-releasing hormone and c-Fos coexpression in middle-aged female mice.

Hypothalamic glutamate (Glu) and γ-GABA neurotransmission are involved in the ovarian hormone-induced gonadotrophin-releasing hormone (GnRH)/luteinising hormone (LH) surge in rodents. Studies have shown that reduced Glu and increased γ-GABA in the rostral preoptic area (rPOA) of the hypothalamus, where most activated GnRH neurones are located, play a key role in decreasing the reproductive function of female rats. However, the mechanism underlying the altered balance of these neurotransmitters is poorly understood. In the present study, we observed a decline in the function of GnRH neurones in the rPOA at the time of the GnRH/LH surge in middle-aged intact female mice with regular oestrous cycles. In young mice, there is an increase of vesicular Glu transporter 2 on the pro-oestrus afternoon, which is not observed in middle-aged mice. By contrast, vesicular γ-GABA transporter levels in young mice decrease at the time of the LH surge, whereas they increase in middle-aged mice. Of note, we found that, in middle-aged mice at the time of the GnRH/LH surge, the phosphorylation of synapsin I at Ser603 and Ca(2+) /calmodulin-dependent kinase IIα was significantly lower than in young mice. These data suggest that, in middle-aged mice, higher levels of presynaptic stores of GABA, a lack of increase of Glu and a decreased ability of synaptic vesicle mobilisation could account for the imbalance of Glu and GABA in the rPOA, which decreases the activation of GnRH neurones.