Vagus nerve stimulation as a promising neuroprotection for ischemic stroke via α7nAchR-dependent inactivation of microglial NLRP3 inflammasome.

Ischemic stroke is a major cause of disability and death worldwide, and its management requires urgent attention. Previous studies have shown that vagus nerve stimulation (VNS) exerts neuroprotection in ischemic stroke by inhibiting neuroinflammation and apoptosis. In this study, we evaluated the timing for VNS intervention in ischemic stroke, and the underlying mechanisms  of VNS-induced neuroprotection. Mice were subjected to transient middle cerebral artery occlusion (tMCAO) for 60 min. The left vagus nerve at cervical level was exposed and attached to an electrode connected to a low-frequency electrical stimulator. Vagus nerve stimulation (VNS) was given for 60 min before, during and after tMCAO (Pre-VNS, Dur-VNS, Post-VNS). Neurological function was assessed 24 h after reperfusion. We found that all the three VNS significantly protected against the tMCAO-induced injury evidenced by improved neurological function and reduced infarct volume. Moreover, the Pre-VNS was the most effective against the ischemic injury. We found that tMCAO activated microglia in the ischemic core and penumbra regions of the brain, followed by the NLRP3 inflammasome activation-induced neuroinflammation, which finally triggered neuronal death. VNS treatment preserved α7nAChR expression in the penumbra regions, inhibited NLRP3 inflammasome activation and ensuing neuroinflammation, rescuing cerebral neurons. The role of α7nAChR in microglial NLRP3 inflammasome activation in ischemic stroke was further validated using genetic manipulations, including Chrna7 knockout mice and microglial Chrna7 overexpression mice, as well as pharmacological interventions using the α7nAChR inhibitor methyllycaconitine and agonist PNU-282987. Collectively, this study demonstrates the potential of VNS as a safe and effective strategy to treat ischemic stroke, and presents a new approach targeting microglial NLRP3 inflammasome, which might be therapeutic for other inflammation-related diseases.

Synthesis and anti-HIV activity of non-nucleoside reverse-transcriptase inhibitor DB02 phosphate derivatives based on water-soluble optimization.

To improve the water solubility of anti-human immunodeficiency virus (HIV) agent DB02, an excellent non-nucleoside reverse-transcriptase inhibitor (NNRTI) obtained in our previous efforts, we designed and synthesized four phosphate derivatives of DB02 based on the molecular model of DB02 with RT. Here, the antiviral activity of these four derivatives was detected, leading to the discovery of compound P-2, which possessed a superior potency to the lead compound DB02 against wild-type HIV-1 and a variety of HIV-resistant mutant viruses significantly. Furthermore, the water solubility of P-2 was nearly 17 times higher than that of DB02, and the pharmacokinetic test in rats showed that P-2 demonstrate significantly improved oral bioavailablity of 14.6%. Our study showed that the introduction of a phosphate ester group at the end of the C-2 side chain of DB02 was beneficial to the improvement of its antiviral activity and pharmacokinetic properties, which provided a promising lead for the further development of S-DACOs type of NNRTIs.

C6-structural optimizations of 2-aryl-1H-pyrazole-S-DABOs: From anti-HIV to anti-DENV activity.

Both HIV and DENV are serious threats to human life, health and social economy today. So far, no vaccine for either HIV or DENV has been developed successfully. The research on anti-HIV or DENV drugs is still of great significance. In this study we developed a series of novel 2-Aryl-1H-pyrazole-S-DABOs with C6-strucutral optimizations as potent NNRTIs, among which, 8 compounds had low cytotoxicity and EC50 values in the range of 0.0508 âˆ¼ 0.0966 µM, and their selectivity index was SI > 1415 âˆ¼ 3940. In particular, two compounds 4a and 4b were identified to have good inhibitory effects on DENV of four serotypes. The EC50 of compound 4a and 4b against DENV-II (13.2 µM and 9.23 µM, respectively) were better than that of the positive control ribavirin (EC50 = 40.78 µM). In addition, the effect of C-6 substituents on the anti-HIV or anti-DENV activity of these compounds was also discussed.

CYP17A1 gene mutations and hypertension variations found in 46, XY females with combined 17α-hydroxylase/17, 20-lyase deficiency.

The aim of this study was to analyze the structural consequences of the mutations in CYP17A1 gene and their relationship with the variations of clinical manifestations in three patients who presented with complete or partial combined 17α-hydroxylase/17,20-lyase deficiency (17OHD). DNA sequences of the coding exons and intron/exon boundaries of the CYP17A1 gene were analyzed for mutations. In silico analysis with computational three-dimensional model of human P450c17 and multiple alignments analysis were performed to evaluate the spatial conformational changes by missense mutations. Five mutations p.S117fs (c.351_352delCT), p.H373L (c.1184 A>T), p.Y329fs (c.985_987delTACinsAA), p.A82D (c.245 C>A) and p.L209P (c.626 T>C) were identified in three patients, respectively. The novel mutation p.S117fs (c.351_352delCT) has not been reported previously. In silico analysis explained the conformational changes by the described mutations, which resulted in different severe 17OHD. Our studies also suggest that molecular data accompanying with in silico analysis of the CYP17A1 gene are much helpful for the diagnosis, management and genetic counseling of 17OHD.

Carnosol ameliorated cancer cachexia-associated myotube atrophy by targeting P5CS and its downstream pathways.

Introduction: Carnosol exhibited ameliorating effects on muscle atrophy of mice developed cancer cachexia in our previous research. Method: Here, the ameliorating effects of carnosol on the C2C12 myotube atrophy result from simulated cancer cachexia injury, the conditioned medium of the C26 tumor cells or the LLC tumor cells, were observed. To clarify the mechanisms of carnosol, the possible direct target proteins of carnosol were searched using DARTS (drug affinity responsive target stability) assay and then confirmed using CETSA (cellular thermal shift assay). Furthermore, proteomic analysis was used to search its possible indirect target proteins by comparing the protein expression profiles of C2C12 myotubes under treatment of C26 medium, with or without the presence of carnosol. The signal network between the direct and indirect target proteins of carnosol was then constructed. Results: Our results showed that, Delta-1-pyrroline-5-carboxylate synthase (P5CS) might be the direct target protein of carnosol in myotubes. The influence of carnosol on amino acid metabolism downstream of P5CS was confirmed. Carnosol could upregulate the expression of proteins related to glutathione metabolism, anti-oxidant system, and heat shock response. Knockdown of P5CS could also ameliorate myotube atrophy and further enhance the ameliorating effects of carnosol. Discussion: These results suggested that carnosol might ameliorate cancer cachexia-associated myotube atrophy by targeting P5CS and its downstream pathways.

N-(4-Bromo-phen-yl)-2-[(1-cyclo-hexyl-meth-yl-1H-1,2,4-triazol-3-yl)sulfanyl]-acetamide.

The title compound, C(17)H(21)BrN(4)OS, was synthesized as a potential reverse transcriptase (RT) inhibitor of the human immunodeficiency virus type 1 (HIV-1). In the molecule, there is an N-H⋯S hydrogen bond making a five-membered ring. In the crystal, mol-ecules are connected into centrosymmetric dimers via pairs of N-H⋯N and weak C-H⋯N hydrogen bonds. The crystal structure also features C-H⋯O inter-actions.

Impact of RET Screening on the Management of Multiple Endocrine Neoplasia Type 2A: 10 Years Experience and Follow-Up in Three Families.

BACKGROUND: Multiple endocrine neoplasia type 2A (MEN 2A) is mainly caused by germline RET codon C634 mutation and is characterized by Medullary Thyroid Carcinoma (MTC), pheochromocytoma (PHEO), and hyperparathyroidism (HPTH). The early diagnosis and initial normative treatment are helpful for the long-term outcome of MEN2A. METHODS: Three index cases and their 29 relatives from three families with MEN2A were included in this study. Genetic screening was performed on all participants. Demographic, clinical profiles, tumor histopathologic features, and follow-up records were systematically analyzed. RESULTS: In total, RET C634Y mutation was identified in 10 individuals (10/32, 31.3%). Among them, 5 presented with MTC symptoms, whereas the other 5 did not show apparent clinical manifestation, and all were subjected to thyroidectomy with varying neck dissection. Compared to individuals in the former, the latter benefited greatly from RET screening with significantly younger age at diagnosis of MTC and surgery (18.1 ± 13.8 years vs. 39.0 ± 14.1 years, P =0.045), and lessaggressive MTC behavior (size: 0.74 vs. 2.82 cm, P =0.026; LN+/resected: 20.0% vs. 100.0%, P =0.048) and also lower recurrence rate of MTC (20.0% vs. 100.0%, P =0.048). The PHEO was identified in 6 of the 10 carriers (60.0%), and all had undergone adrenal-sparing surgery. During the 10 years of follow-up, one (16.7%) developed recurrence of PHEO. CONCLUSION: Integrated RET screening, serum calcitonin, and plasma metanephrine/ normetanephrine levels can facilitate the early diagnosis and standardized MTC/PHEO surgery to improve the prognosis of MEN2A. Laparoscopic adrenal-sparing surgery prior to the bilateral total thyroidectomy is a preferred surgical approach for PHEO.

Identification of 6ω-cyclohexyl-2-(phenylamino carbonylmethylthio)pyrimidin-4(3H)-ones targeting the ZIKV NS5 RNA dependent RNA polymerase.

Zika virus (ZIKV), a mosquito-borne flavivirus, is a global health concern because of its association with severe neurological disorders such as neonatal microcephaly and adult Guillain-Barre syndrome. Although many efforts have been made to combat ZIKV infection, there is currently no approved vaccines or antiviral drugs available and there is an urgent need to develop effective anti-ZIKV agents. In this study, 26 acetylarylamine-S-DACOs derivatives were prepared, and eight of them were found to have inhibitory activity against Zika virus. Among these substances, 2-[(4-cyclohexyl-5-ethyl-6-oxo-1,6-dihydropyrimidin-2-yl)thio]-N-(3,5-difluorophenyl)acetamide (4w) with the best anti-ZIKV activity was selected for in-depth study of antiviral activity and mechanism of action. Here, we discovered 4w targeted on the ZIKV NS5 RNA -dependent RNA polymerase (RdRp), which exhibited good in vitro antiviral activity without cell species specificity, both at the protein level and at the RNA level can significantly inhibit ZIKV replication. Preliminary molecular docking studies showed that 4w preferentially binds to the palm region of NS5A RdRp through hydrogen bonding with residues such as LYS468, PHE466, GLU465, and GLY467. ZIKV NS5 RdRp enzyme activity experiment showed that 4w could directly inhibit ZIKV RdRp activity with EC50 = 11.38 ± 0.51 µM. In antiviral activity studies, 4w was found to inhibit ZIKV RNA replication with EC50 = 6.87 ± 1.21 µM. ZIKV-induced plaque formation was inhibited with EC50 = 7.65 ± 0.31 µM. In conclusion, our study disclosed that acetylarylamine-S-DACOs is a new active scaffolds against ZIKV, among which compound 4w was proved to be a potent novel anti-ZIKV compound target ZIKV RdRp protein. These promising results provide a future prospective for the development of ZIKV RdRp inhibitors.

Thermal Analysis of Blood Flow Alterations in Human Hand and Foot Based on Vascular-Porous Media Model.

Microvascular and Macrovascular diseases are serious complications of diabetic mellitus, which significantly affect the life quality of diabetic patients. Quantitative description of the relationship between temperature and blood flow is considerably important for non-invasive detection of blood vessel structural and functional lesions. In this study, thermal analysis has been employed to predict blood flow alterations in a foot and a cubic skin model successively by using a discrete vessel-porous media model and further compared the blood flows in 31 diabetic patients. The tissue is regarded as porous media whose liquid phase represents the blood flow in capillaries and solid phase refers to the tissue part. Discrete vascular segments composed of arteries, arterioles, veins, and venules were embedded in the foot model. In the foot thermal analysis, the temperature distributions with different inlet vascular stenosis were simulated. The local temperature area sensitive to the reduction of perfusion was obtained under different inlet blood flow conditions. The discrete vascular-porous media model was further applied in the assessment of the skin blood flow by coupling the measured skin temperatures of diabetic patients and an inverse method. In comparison with the estimated blood flows among the diabetic patients, delayed blood flow regulation was found in some of diabetic patients, implying that there may be some vascular disorders in these patients. The conclusion confirms the one in our previous experiment on diabetic rats. Most of the patients predicted to be with vascular disorders were diagnosed as vascular complication in clinical settings as well, suggesting the potential applications of the vascular-porous media model in health management of diabetic patients.

[Analysis of mandibular canal in patients with prognathism and its clinical significance].

PURPOSE: The aim of this study was to investigate the position and course of mandibular canal in patients with prognathism using cone-beam CT(CBCT)，and explore its clinical significance. METHODS: The mandibles of 28 patients with skeletal Class Ⅲ prognathism (10 males and 18 females) and 20 normal people(12 males and 8 females) were analyzed on CBCT. The position and course of mandibular canal from the lowest point of mandibular foramen to the first molar were measured at five specific sections from the mandibular foramen to the mandibular first molar area, including inner and outer diameter, mandibular bone thickness ,distance between mandibular canal and distance to the buccal and lingual bone cortex, and the inferior border of mandible bone. Statistical analysis was performed using SPSS 17.0 software package. RESULTS: The mandibular bones of patients with prognathism gradually thickened from the mandibular ramus to the forepart of the mandibular angle and thinned forward to the position where the mandibular first molar projected on the mandibular bone. Cortical bone of buccal side of the mandibular ramus thickened forward to the mandibular angle and thinned to the mandibular body. The width of buccal bone marrow cavity and diameter of mandibular bone were statistically different between genders. CONCLUSIONS: This study found that the difference between the thickness of the mandibular ramus in patients with mandibular protrusion from the controls was mainly due to the difference between the width of the buccal bone marrow cavity. CBCT can accurately locate the mandibular canal and display anatomic structures.

Reverse facial-submental artery island flap for the reconstruction of maxillary defects after cancer ablation.

This study assessed the reliability of the reverse facial-submental artery island flap for reconstructing maxillary defects. Twelve patients with cancer underwent surgical resection and sequential maxillary reconstruction using a reverse facial-submental artery island flap. There were 9 men and 3 women. The patients ranged in age from 42 to 73 years. Palate squamous cell carcinoma was present in 7 cases; and maxillary gingival squamous cell carcinoma, in 5 cases. The remaining defects were classified as class 1 in one case, class 2a in 8 cases, and class 3b in 3 cases. The sizes of the skin paddle varied from a minimum of 4 x 12 cm to a maximum of 5 x 12 cm. No flaps failed. There were no donor-site problems. The patients were followed up for 16 to 30 months; one case of tumor local recurrence was observed. The reverse facial-submental artery island flap is safe, quick, and simple to elevate. The flap can be used reliably for reconstructing maxillary defects.

[The investigation of the relationship between the phenotypes of 46, XX males and the SRY gene].

OBJECTIVE: To investigate the relationship between the phenotypes in XX male patients and the sex determining region(SRY) gene. METHODS: Multiple polymerase chain reactions were carried out in 6 male patients with karyotype of 46, XX, and then the PCR products were sequenced directly. RESULTS: Three cases of male infertility were positive for the SRY gene without evident malformation in their extra genitalia, while 3 cases with testes were negative for the SRY gene, with evident malformation in their extra genitalia. CONCLUSION: The SRY gene is key in sex determination and development, yet there might be other important genes involved.

Synthesis and biological evaluation of novel 6-substituted 5-alkyl-2-(arylcarbonylmethylthio)pyrimidin-4(3H)-ones as potent non-nucleoside HIV-1 reverse transcriptase inhibitors.

A novel series of 2-arylcarbonylmethylthio-6-arylmethylpyrimidin-4(3H)-ones have been synthesized and evaluated for in vitro anti-HIV activities in MT-4 cells. Most of these new compounds showed moderate to potent activities against wild-type HIV-1 with an EC(50) range from 8.97 microM to 0.010 microM. Among them, the 6-(3,5-dimethylbenzyl) analogue 5p was identified as the most promising compound (EC(50)=0.010 microM, SI>31,800) associated with moderate activity against the HIV-1 double mutant RT strain K103N+Y181C. The structure-activity relationships of these new congeners were further discussed.

[Perioperative nursing of zygomatic implant surgery for patients with severe periodontitis].

PURPOSE: To analyze the perioperative nursing characteristics about severe periodontitis patients with immediate zygomatic implantation, and to summarize the clinical nursing experience. METHODS: Preoperatively, all patients received psychological counseling, oral hygiene instruction, and periodontal care with necessary means after physical examination. Cooperative procedures were preformed between nurses and surgeon during operation. After operation, nursing interventions were given including wound care, pain control, ice compression, appropriate diet and close follow-up. RESULTS: All patients were successfully operated without serious complications. Favorable implant primary stability was achieved in each patient. No patient developed wound dehiscence, one patient had wound pain and one patient had minor facial swelling. Temporary restoration was conducted to restore occlusal relationship and masticatory function within 48 hours. CONCLUSIONS: Comprehensive nursing intervention during perioperative period is necessary and deserves to be applied for patients with severe periodontitis who are treated with immediate zygomatic implant surgery.

[Retrospective study on postoperative complications and nursing measure for the oldest old patients receiving free flap reconstruction with head and neck cancers].

PURPOSE: To analyze the postoperative complications and nursing measures for the oldest old patients with head and neck cancer, who received free flap reconstruction. METHODS: A retrospective study for oldest old patients treated in the department of Oromaxillofacial Head and Neck Oncology of our hospital from 2005 to 2015 was conducted. All patients with head and neck cancer received free flap reconstruction with age≥85 years were selected. Demographics, clinical, nursing information and prognosis were collected. SPSS 17.0 software package was used for data analysis. RESULTS: There were a total of 24 oldest old patients, among whom 15 were male and 9 were female. The median age was 86 years old, 20 patients had co-morbidity, 18 patients had ASA Ⅱ and 6 patients had ASA Ⅲ. The median operation time was 420 minutes, while the median intraoperative blood loss was 550 mL. 16 patients underwent tracheotomy, 9 patients suffered from postoperative surgical complications and 16 patients experienced postoperative medical complications. CONCLUSIONS: The oldest old patients with head and neck cancer have high possibility of postoperative complications. Preoperative assessment, accurate postoperative observation and nursing measures can reduce the negative impact of postoperative complications and improve prognosis effectively.

[The mechanism of root hair development and molecular regulation in plants].

The formation of the root epidermis in Arabidopsis thaliana provides a simple model to study mechanisms underlying patterning in plants. Root hair increases the root surface area and effectively increases the root diameter, so root hair is thought to aid plants in nutrient uptake, anchorage and microbe interactions. The determination of root hair development has two types, lateral inhibition with feedback and position-dependent pattern of cell differentiation. The initiation and development of root hair in Arabidopsis provide a simple and efficacious model for the study of cell fate determination in plants. Molecular genetic studies identify a suite of putative transcription factors which regulate the epidermal cell pattern. The homeodomain protein GLABRA2 (GL2), R2R3 MYB-type transcription factor WEREWOLF (WER) and WD-repeat protein TRANSPARENTT TESTA GLABRA (TTG) are required for specification of non-hair cell type. The CAPRICE (CPC) and TRYPTICHON (TRY) are involved in specifying the hair cell fate.

Chromosome 13q deletion syndrome involving 13q31­qter: A case report.

Partial deletions on the long arm of chromosome 13 lead to a number of different phenotypes depending on the size and position of the deleted region. The present study investigated 2 patients with 13q terminal (13qter) deletion syndrome, which manifested as anal atresia with rectoperineal fistula, complex type congenital heart disease, esophageal hiatus hernia with gastroesophageal reflux, facial anomalies and developmental and mental retardation. Array comparative genomic hybridization identified 2 regions of deletion on chromosome 13q31­qter; 20.38 Mb in 13q31.3­qter and 12.99 Mb in 13q33.1­qter in patients 1 and 2, respectively. Comparisons between the results observed in the present study and those obtained from patients in previous studies indicate that the gene encoding ephrin B2 (EFNB2) located in the 13q33.3­q34 region, and the gene coding for endothelin receptor type B, in the 13q22.1­31.3 region, may be suitable candidate genes for the observed urogenital/anorectal anomalies. In addition, the microRNA­17­92a­1 cluster host gene and the glypican 6 gene in the 13q31.3 region, as well as EFNB2 and the collagen type IV a1 chain (COL4A1) and COL4A2 genes in the 13q33.1­q34 region may together contribute to cardiovascular disease development. It is therefore possible that these genes may be involved in the pathogenesis of complex type congenital heart disease in patients with 13q deletion syndrome.

[Morphometric analysis of maxillofacial bone in 48 patients with ectodermal dysplasia].

PURPOSE: The study was performed to analyze the maxillofacial morphology of 48 patients with ectodermal dysplasia (ED) and to provide some reference for implant treatment of ED patients. METHODS: The study collected the clinical data and lateral cephalometric radiographs of 48 patients with ED treated between June 2013 and October 2016. The patients were grouped as follows: group 1, 23 patients, at least with two kinds of ectodermal disorder, besides hypodontia; group 2, 25 patients, only with one kind of ectodermal disorder, besides hypodontia. The number of missing teeth and maxillofacial cephalometric parameters were compared between 2 groups. SPSS 17.0 software package was performed for statistical analysis. RESULTS: The number of missing teeth was significantly larger in group 1(22.3±5.8) than group 2 (12±6.6) (P<0.05). The number of retained deciduous teeth was significantly larger in group 1 (8±4) than in group 2 (2.4±2.7) in females (P<0.05). Cephalometric analysis showed a reduced maxilla length (ANS-Ptm), retrusive maxilla (S-Ptm, SNA, NA-PA), a normal size (S-Co) and length (Co-Po) of mandible, a protruding mandible (NP-FH) and chin (Y axis), skeletal Angle Ⅲ malocclusion(ANB<0) and a reduced facial height (N-Me) in two groups. SNA, NA-PA, ANS-Ptm, S-Ptm and Y axis were significantly decreased and NP-FH was significantly increased in group 1 compared to group 2 in males (P<0.05). CONCLUSIONS: Maxillofacial morphology of ED patients typically presented a reduced maxilla length and retrusive maxilla, a normal size and length of mandible, a protruding mandible and chin, skeletal Angle Ⅲ malocclusion and a reduced facial height. The severity of abnormalities in maxillofacial bone morphology was correlated positively with absence of permanent teeth. Furthermore, retained deciduous teeth might have positive effect on the development of maxillofacial bone.

CNTNAP2 Is Significantly Associated With Speech Sound Disorder in the Chinese Han Population.

Speech sound disorder is the most common communication disorder. Some investigations support the possibility that the CNTNAP2 gene might be involved in the pathogenesis of speech-related diseases. To investigate single-nucleotide polymorphisms in the CNTNAP2 gene, 300 unrelated speech sound disorder patients and 200 normal controls were included in the study. Five single-nucleotide polymorphisms were amplified and directly sequenced. Significant differences were found in the genotype (P = .0003) and allele (P = .0056) frequencies of rs2538976 between patients and controls. The excess frequency of the A allele in the patient group remained significant after Bonferroni correction (P = .0280). A significant haplotype association with rs2710102T/+rs17236239A/+2538976A/+2710117A (P = 4.10e-006) was identified. A neighboring single-nucleotide polymorphism, rs10608123, was found in complete linkage disequilibrium with rs2538976, and the genotypes exactly corresponded to each other. The authors propose that these CNTNAP2 variants increase the susceptibility to speech sound disorder. The single-nucleotide polymorphisms rs10608123 and rs2538976 may merge into one single-nucleotide polymorphism.

[Transmission disequilibrium test for congenital dislocation of the hip and HOXB9 gene or COL1AI gene].

OBJECTIVE: To detect the correlation between the congenital dislocation of the hip (CDH) and HOXB9 gene or COL1AI gene. METHODS: A microsatellite DNA marker D17S1820 was chosen in the region of chromosome 17q21 where exists the HOXB9 gene which regulates the embryonic limb development and exists the COL1AI gene. The genotypes of 303 members in 101 CDH nuclear family trios were analyzed by the techniques of polymerase chain reaction(PCR) and denaturing polyacrylamide gel electrophoresis. Then transmission disequilibrium test (TDT) was used to test the data of genotypes. RESULTS: There exist 12 alleles at this polymorphic locus. Transmission disequilibrium was found between CDH and the fourth allele of D17S1820 (chi-square=6.025,P=0.014). CONCLUSION: CDH is associated with the region of chromosome 17q21. HOXB9 gene and/or COL1AI gene may be susceptibility genes of CDH.