Mechanisms of PiT2-loop7 Missense Mutations Induced Pi Dyshomeostasis / 神经科学通报·英文版

PiT2 is an inorganic phosphate (Pi) transporter whose mutations are linked to primary familial brain calcification (PFBC). PiT2 mainly consists of two ProDom (PD) domains and a large intracellular loop region (loop7). The PD domains are crucial for the Pi transport, but the role of PiT2-loop7 remains unclear. In PFBC patients, mutations in PiT2-loop7 are mainly nonsense or frameshift mutations that probably cause PFBC due to C-PD1131 deletion. To date, six missense mutations have been identified in PiT2-loop7; however, the mechanisms by which these mutations cause PFBC are poorly understood. Here, we found that the p.T390A and p.S434W mutations in PiT2-loop7 decreased the Pi transport activity and cell surface levels of PiT2. Furthermore, we showed that these two mutations attenuated its membrane localization by affecting adenosine monophosphate-activated protein kinase (AMPK)- or protein kinase B (AKT)-mediated PiT2 phosphorylation. In contrast, the p.S121C and p.S601W mutations in the PD domains did not affect PiT2 phosphorylation but rather impaired its substrate-binding abilities. These results suggested that missense mutations in PiT2-loop7 can cause Pi dyshomeostasis by affecting the phosphorylation-regulated cell-surface localization of PiT2. This study helps understand the pathogenesis of PFBC caused by PiT2-loop7 missense mutations and indicates that increasing the phosphorylation levels of PiT2-loop7 could be a promising strategy for developing PFBC therapies.

The Pathology of Primary Familial Brain Calcification: Implications for Treatment / 神经科学通报·英文版

Primary familial brain calcification (PFBC) is an inherited neurodegenerative disorder mainly characterized by progressive calcium deposition bilaterally in the brain, accompanied by various symptoms, such as dystonia, ataxia, parkinsonism, dementia, depression, headaches, and epilepsy. Currently, the etiology of PFBC is largely unknown, and no specific prevention or treatment is available. During the past 10 years, six causative genes (SLC20A2, PDGFRB, PDGFB, XPR1, MYORG, and JAM2) have been identified in PFBC. In this review, considering mechanistic studies of these genes at the cellular level and in animals, we summarize the pathogenesis and potential preventive and therapeutic strategies for PFBC patients. Our systematic analysis suggests a classification for PFBC genetic etiology based on several characteristics, provides a summary of the known composition of brain calcification, and identifies some potential therapeutic targets for PFBC.

The Research of Monoclonal Antibody KMP1 Inhibited Bladder Cancer EJ Cell Lines Growth and Metastasis in Vivo / 昆明医科大学学报

Objective To research the monoclonal antibody KMP1 inhibited bladder cancer EJ cell lines growth and metastasis in vivo by bioluminescence imaging. Methods Immunohistochemistry was used to determine the KMP1 binding to EJ and EJ-GFP cell lines. The xenograft tumor cell growth and distribution were measured by vernier calipers and dynamic in vivo fluorescence imaging. Immunohistochemistry and H&E counterstaining researched the feature of the xenograft tumor. Results Cell growth curves of EJ and EJ-GFP cells were similar. EJ-GFP had a green fluorescence. In EJ-GFP nude mouse tumor model, the addition of KMP1 significantly inhibited tumor growth and extended the average life span of nude mice. Both EJ and EJ-GFP cells can bind to KMP1,and the weight of transplanted tumors in the KMP1 treatment group was significantly lower than that of the mIgG control group (P＜0.001).Conclusion KMP1 has a promising antitumor effect in vivo. It might be valuable for development as a promising targeted agent for bladder cancer.

Repeated superovulation induction after failure in previous IVF-ET cycles with the ultra-long protocol: Analysis of outcomes of clinical pregnancy / 中华男科学杂志

Objective@#To analyze the clinical outcomes of repeated superovulation induction in patients with adenomyosis or moderate to severe pelvic endometriosis after failure in previous IVF-ET cycles with the ultra-long protocol.@*METHODS@#We retrospectively analyzed the clinical data about 37 patients with adenomyosis or moderate to severe pelvic endometriosis in our center from 2009 to 2013, who underwent repeated IVF-ET after failure in the previous cycles with the ultra-long protocol, namely by injection of 2－6 ampoules of 3.75 mg gonadotropin-releasing hormone agonist (GnRH-a). All the patients met the following requirements: hCG-negative at 14 days after transfer, within 3－7 days after menstruation, and properly down-regulated serum follicle stimulating hormone (FSH) (<10 mIU/ml), luteinizing hormone (LH) (<10 mIU/ml), estradiol (E2) (<30 pg/ml), follicle diameter (<10 mm) and endometrial thickness, and received GnRH (Gonal-F, Serono) for ovulation induction. We compared the clinical and laboratory data and pregnancy outcomes between the first and repeated cycles before and after ovulation induction.@*RESULTS@#The repeated cycles, as compared with previous ones, showed significant increases in the antral follicle count (AFC) on the first day of stimulation (7.55 ± 1.86 vs 6.45 ± 2.5, P<0.05), number of follicles =≥14 mm in diameter on the hCG trigger day (7.81 ± 3.6 vs 5.56 ± 3.68, P<0.05), level of E2 (［2 362.15 ± 1 210.49］ vs ［1 749.22 ± 1 139.44］ pg/ml, P<0.05), and numbers of oocytes retrieved (7.51 ± 3.23 vs 4.78 ± 3.41, P<0.05) and embryos transferred (2.00 ± 0.33 vs 1.50 ± 0.67, P<0.05), exhibited a remarkably reduction in the dose of GnRH (［1 791.65 ± 1 889.41］ vs ［3 439.56 ± 1 836.53］ IU, P<0.05), and achieved a clinical pregnancy rate of 62.16%.@*CONCLUSIONS@#With proper reduction of the FSH, LH and E2 levels and follicle diameter, repeated superovulation induction for IVF-ET can improve the ovarian response and pregnancy outcomes of the patients with adenomyosis or moderate to severe pelvic endometriosis after failure in the previous IVF-ET cycles with the ultra-long protocol.

3.0 T MRI in analysis of atherosclerotic middle cerebral artery stenosis / 第二军医大学学报

Objective To apply 3.0 T MRI(3-dimensional time-of-flight and high-resolution MRI vessel wall imaging) technique for analyzing atherosclerotic middle cerebral stenosis, location of the plaque and remodeling of the middle cerebral artery. Methods 3.0 T MRI was used to examine 64 patients with middle cerebral atherosclerotic stenosis, determining the plaque location and thickness. The plaque area ratio was defined as plaque area/cross-sectional area of the vessel (VA); remodeling ratio was detined as VA/average of the proximal and distal normal artery; and the luminal area of the vessil (LA) ratio was delined as LA/average of the proximal and distal normal artery. Results The atherosclerotic plaque of middle cerebral artery (M1 segment) most frequently formed annular plaque and was located at the ventral and inferior wall, with the plaque thickness being 0.5-1.5 mm (72.5%). The middle cerebral atherosclerotic plaque remodeling modes were mainly positive (outward) remodeling. Conclusion 3.0 T MRI can help to demonstrate the distribution of the middle cerebral atherosclerotic plaque and the remodeling of the middle cerebral artery, which is worth further studying.

The infection status and epidemic rule of new bunia virus in the livestock and poultry in hilly area of Jiaodong peninsula / 中华预防医学杂志

<p><b>OBJECTIVE</b>To understand the infection status and epidemic rule of new bunia virus in the livestock and poultry which are closely related with humans such as sheep, cattle, dogs, pigs and chicken in the hilly area of Jiaodong peninsula in Shandong province.</p><p><b>METHODS</b>Penglai and Laizhou in the hilly area of Jiaodong peninsula in Shandong province where severe fever with thrombocytopenia syndrome cases occurred in 2010 were selected as experimental sites. During April to November in 2011, serum specimens of the sheep, cattle, dogs, pigs and chicken with ticks in endemic area were randomly collected by random number table.5 ml venous blood was collected in each livestocks or poultries and there were total 3576 samples.New bunia virus antibody in different species of livestocks or poultries serum was continuously detected using double antigen sandwich enzyme-linked immunosorbent assay, and the infection rates of new bunia virus between different species of livestocks or poultries and between Penglai and Laizhou were analyzed using chi-square test.</p><p><b>RESULTS</b>Test results in 3576 samples of livestocks or poultries serum specimen showed that the infection rate was as high as 63% (636/1013) in sheep, 53% (444/841)in cattle, 46% (242/530) in chicken, 29% (104/362)in the dogs, and 1% (12/830) in pigs. There were significant differences of new bunia virus infection among different species (χ(2) = 815.26, P < 0.05).In Penglai, the infection rate was as high as 71% (400/563) in sheep, 57% (232/409)in cattle, 35% (93/266) in chicken, 44% (796/1819)in total, while in Laizhou, the infection rate was 53% (236/450)in sheep, 49% (212/432)in cattle, 56% (149/264)in chicken, 36% (642/1757)in total, their difference was statistically significant(χ(2) values were 37.04, 4.93, 24.63, 19.38, all P values were < 0.05).Infection rates of dogs and pigs showed no obvious fluctuation.However, there were two peaks of infection in sheep in summer and autumn, the infection rate was as high as 62% (68/110) in June and 86% (204/236) in November;There were two peaks of infection in cattle in spring and autumn, the infection rate was as high as 56% (53/94) in April and 73% (116/159) in November; there was only one peak of infection in chicken, the infection rate was as high as 65% (55/85) in September.</p><p><b>CONCLUSION</b>The infection rate is higher in sheep, cattle, chickens and dogs in the hilly area of Jiaodong peninsula. The peak season is spring, summer and autumn.</p>

Appropriate prolongation of GnRH-a down-regulation improves the synchronism of follicular development / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the relatively appropriate actuation time for ovarian super-stimulation of IVF-ET by comparing the influences of different down-regulation days of chorionic gonadotrophin releasing hormone agonist (GnRH-a) upon the follicular diameter, endometrial thickness and the levels of follicle- stimulating hormone (FSH) , luteinizing hormone (LH) and estradiol (E2).</p><p><b>METHODS</b>We adopted the long protocol of GnRH-a down-regulation in the midluteal phase for 42 patients undergoing IVF-ET. According to the time of GnRH-a down-regulation, we divided the patients into a 10 d, a 15 d and an 18 d group, measured their follicular diameters and endometrial thickness by B-mode ultrasonography, detected the levels of FSH, LH and E2 in the blood, and analyzed the influences of different days of GnRH-a down-regulation on the follicular diameter, endometrial thickness and sexual hormone levels. At 1, 7, 10 and 14 d of down-regulation, we compared the levels of FSH and LH in the blood before the injection of GnRH-a with those 2 and 3 h after it.</p><p><b>RESULTS</b>At 10, 15 and 18 d after down-regulation, the ovarian follicles with the diameter of 3-4 mm accounted for 16.8, 7.09 and 10.38% (P < 0.05, 10 d vs 15 d and 18 d), those with the diameter of 4.5-7.0 mm made up 80.24, 89.55 and 84.62% (P < 0.05, 15 d vs 10 d and 18 d), and those with the diameter of 7.5-10 mm constituted 2.96, 3.36 and 5%, respectively. Endometrial thickness was (7.73 +/- 2.48) mm in the 10 d group, significantly thicker than (5.41 +/- 0.79) mm and (5.24 +/- 0.85) mm in the 15 d and 18 d groups (P < 0.05). The FSH levels in the 10 d, 15 d and 18 d groups were (3.70 +/- 1.10), (3.51 +/- 0.72) and (3.47 +/- 0.61) mIU/ml, the LH levels were (1.23 +/- 1.00), (1.09 +/- 0.47) and (1.22 +/- 0.72) mIU/ml, and the E2 levels were 41.84 +/- 36.81, 32.84 +/- 14.32 and 9.50 +/- 8.23, respectively, with no significant differences among the three groups. At 1, 7, 10 and 14 d of down-regulation, both FSH and LH levels in the blood were increased at 2 and 3 h after GnRH-a injection, most significantly at 1 d (1.87 +/- 1.49 vs 13.33 +/- 7.81 for FSH, 1.06 +/- 1.13 vs 47.40 +/- 29.97 for LH, (P < 0.05).</p><p><b>CONCLUSION</b>In the long protocol of ovarian super-stimulation of IVF-ET, endometrial thickness and the levels of FSH, LH and E2 tended to be stable at 10 d of GnRH-a down-regulation. The percentage of the follicles with the diameter of 4.5-7.0 mm was higher at 15 d than at 10 d, but rose no more at 18 d except for an increased number of smaller follicles 3-4 mm in diameter. Therefore, appropriate prolongation of GnRH-a down-regulation can improve the synchronism of follicular development.</p>

Identification of VEGFR3 gene mutation in a Chinese family with autosomal dominant primary congenital lymphoedema / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify the disease-causing gene in a four-generation Chinese family with 9 members affected with primary congenital lymphoedema (PCL, also known as Milroy disease).</p><p><b>METHODS</b>Linkage analysis was performed with a few microsatellite markers flanking the candidate genetic loci for PCL, including 3 known genes associated with autosomal dominant PCL. For mutation analysis, VEGFR3 gene was sequenced with DNA from the proband. Direct DNA sequencing of exon 25 of the VEGFR3 gene was performed in all family members.</p><p><b>RESULTS</b>The disease gene in the family was mapped to chromosome 5q35.3 with a maximum Lod score of 2.07. Direct DNA sequencing of VEGFR3 gene revealed a heterozygous C to T transition at nucleotide 3341, resulting in p.Pro1114Leu mutation. The p.Pro1114Leu mutation co-segregated with all affected individuals in the family.</p><p><b>CONCLUSION</b>This study identified a C3341T (p.Pro1114Leu) mutation in the VEGFR3 gene in a Chinese family with PCL, provided evidence that VEGFR3 mutation can cause PCL in Chinese.</p>

The G401D mutation of OPA1 causes autosomal dominant optic atrophy and hearing loss in a Chinese family / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To describe the clinical and genetic characteristics of a Chinese family affected with optic atrophy 1 (OPA1).</p><p><b>METHODS</b>Linkage analysis and DNA sequencing as well as PCR/restriction fragment length polymorphism (RFLP) analysis were performed to identify the disease-causing mutation.</p><p><b>RESULTS</b>A missense mutation, G401D in the OPA1 gene was identified, and the patients demonstrate inherited syndrome of optic atrophy and hearing loss.</p><p><b>CONCLUSION</b>The present study demonstrates that a mutation in the OPA1 gene can cause optic atrophy in Chinese patients, and supports the notion that OPA1 mutation may lead to OPA1 combined with hearing loss.</p>

Two novel missense mutations of GATA4 gene in Chinese patients with sporadic congenital heart defects / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify mutations in GATA4 gene in Chinese patients with sporadic congenital heart defects (CHD).</p><p><b>METHODS</b>Single stranded conformation polymorphism (SSCP) analysis was performed to screen for mutations in all six exons and exon-intron boundaries of GATA4 in 31 individuals with CHD. Direct DNA sequencing was used to identify the specific mutations.</p><p><b>RESULTS</b>Two novel missense mutations, V267M in exon 4, V380M in exon 6, and one polymorphism in intron 6 of GATA4 were identified.</p><p><b>CONCLUSION</b>The above identified two novel GATA4 mutations associated with CHD in Chinese patients. This suggests that the transcription factor GATA4 may play an important role in cardiogenesis.</p>