[Genetic characteristics and clinical analysis of 20 patients with Gaucher's disease].

Gaucher Disease (GD) is an autosomal recessive lysosomal storage disorder characterized by high heterogeneity. This study aimed to further understand the correlation between clinical phenotypes and genotypes in GD patients through a retrospective analysis of 20 cases in Shanxi Bethune Hospital, including their clinical manifestations, laboratory tests, enzyme studies, and genetic results. Among the 20 GD patients, 16 were classified as Type Ⅰ GD with a median age of diagnosis of 24 years, and 4 were classified as Type Ⅲ GD with a median age of diagnosis of 19 years. All patients exhibited splenomegaly and thrombocytopenia, with 16 patients showing skeletal imaging changes, and 5 of them presenting with bone pain symptoms. Genetic analysis revealed 15 distinct mutations, predominantly missense mutations, with L483P being the most prevalent (35.7%), followed by V414L, L303I, and F252I. Mutation sites were predominantly located in exon 7. Noteworthy findings included the first report of the S310G mutation by our research group and the first occurrence of the K196R mutation in the Chinese population. Additionally, the N227S mutation was implicated in a potential association with neuropathy. Despite advancements, Uncertainties still exist in the correlation between clinical phenotypes and genotypes in GD patients.

Global trends in coronary artery disease and artificial intelligence relevant studies: a bibliometric analysis.

OBJECTIVE: Coronary artery disease (CAD) is a major global cause of death, greatly affecting life expectancy and quality of life for populations. With the advent of artificial intelligence (AI), there is new hope for accurately managing CAD. While recent studies have shown remarkable progress in AI and CAD research, there is a gap in comprehensive bibliometric analysis in this field. Therefore, this study aims to provide a thorough analysis of trends and hotspots in AI and CAD-related research utilizing bibliometrics. MATERIALS AND METHODS: Publications on AI and CAD relevant research from 2009 to 2023 were searched through the WoS core database (WoSCC). CiteSpace, VOSviewer and Excel 365 were used to conduct the bibliometric analysis. RESULTS: The bibliometric analysis included 1,248 publications, indicating a steady increase in AI and CAD-related publications annually. The United States of America (USA), China, and Germany were identified as the most influential countries in this field. Research institutions such as Cedars Sinai Med Ctr, Med Univ South Carolina, Harvard Med Sch and Capital Med Univ were the main contributors to research production. FRONT CARDIOVASC MED is the top-ranked journal, while J AM COLL CARDIOL emerged as the most cited journal. Schoepf, U. Joseph, Slomka, Piotr J., Berman, Daniel S. and Dey, Damini were the most prolific authors, while U. Rajendra Acharya was the most frequently co-cited author. Research related to the AI calculation of coronary flow reserve fraction and coronary artery calcification, based on coronary CT to identify CAD and cardiovascular risk, was a key research topic in this field. The potential link between cardiovascular risk stratification and radiomics is currently at the forefront of the field. CONCLUSIONS: This study is the first to use a bibliometric approach to visualize and analyze AI and CAD-related research. The findings provide insights into recent research trends and hotspots in the field and can serve as a reference for scholars to identify critical issues in this field.

Spatiotemporal characteristics of ozone and the formation sensitivity over the Fenwei Plain.

High surface ozone (O3) levels affect human and environmental health. The Fenwei Plain (FWP), one of the critical regions for China's "Blue Sky Protection Campaign", has reported severe O3 pollution. This study investigates the spatiotemporal properties and the causes of O3 pollution over the FWP using high-resolution data from the TROPOspheric Monitoring Instrument (TROPOMI) from 2019 to 2021. This study characterizes spatial and temporal variations in O3 concentration by linking O3 columns and surface monitoring using a trained deep forest machine learning model. O3 concentrations in summer were 2-3 times higher than those found in winter due to higher temperatures and greater solar irradiation. The spatial distributions of O3 correlate with the solar radiation showing decreased trends from the northeastern to the southwestern FWP, with the highest O3 values in Shanxi Province and the lowest in Shaanxi Province. For urban areas, croplands and grasslands, the O3 photochemistry in summer is NOx-limited or in the transitional regime, while it is VOC-limited in winter and other seasons. Reducing NOx emissions would be effective for decreasing O3 levels in summer, while VOC reductions are necessary for winter. The annual cycle in vegetated areas included both NOx-limited and transitional regimes, indicating the importance of NOx controls to protect ecosystems. The O3 response to limiting precursors shown here is of importance for optimizing control strategies and is illustrated by emission changes during the 2020 COVID-19 outbreak.

[Research status and existing problems of lumbar pedicle double-trajectory screw technique].

Lumbar pedicle double-trajectory screw technique refers to the placement of traditional pedicle screw combined with cortical bone trajectory screw in the same pedicle, which has been proved to provide higher fixation strength in osteoporotic vertebral body fixation, and is also a less invasive surgical choice for revision of adjacent segment disease. At present, related basic research and clinical application exploration of double-trajectory screw technique have been carried out, but there are still many problems that need to be further investigated and clarified. The clinical application and existing problems of double screw technique were discussed in this paper for clinical reference.

[Genetic mutation profiles for children with congenital hypothyroidism in Fujian province].

Objective: To explore the mutation characteristics of pathogenic genes in children with congenital hypothyroidism (CH) in Fujian. Methods: The clinical data of 116 unrelated CH children diagnosed in Fujian Provincial Maternal and Child Health Hospital from January 2019 to September 2020 were retrospectively analyzed, including 50 females and 66 males, with an average age of (20±10) days at diagnosis. Targeted exome sequencing technology was used to detect the mutation frequency, type and distribution characteristics of 29 genes related to thyroxine synthesis or thyroid development. Results: Three hundred and fifty-one potential functional mutations were detected in 105 of 116 CH patients, with a detection rate of 90.5% (105/116). DUOX2 (66.4%, 77/116) was the most frequent mutated gene, followed by TG (23.3%, 27/116), DUOXA1 (23.3%, 27/116), and TPO (12.1%, 14/116), which were all involved in thyroid hormone synthesis. Among the 105 children with CH, 70 cases carried double allele mutation. Except for 3 cases of thyroid dysplasia related genes (2 cases of TSHR and 1 case of GLIS3), the rest were also related to thyroid hormone synthesis. The gene with the highest carrier rate was DUOX2 (68.8%, 59/70), followed by TG (8.6%, 6/70), TPO (4.3%, 3/70), DUOXA2 (1.4%, 1/70) and DUOXA1 (1.4%, 1/70). Conclusion: The main mutated genes in CH children in Fujian are the key genes involved in thyroid hormone synthesis, such as DUOX2, TG and TPO.

[Safety and effectiveness of Neuroform Atlas stent-assisted coiling in the endovascular treatment of intracranial aneurysms: a single-center observational study].

Objective: To investigate the safety and effectiveness of Neuroform Atlas stent-assisted coiling in the endovascular treatment of intracranial aneurysms. Methods: This was a retrospective, single-center observational study of 77 patients who underwent endovascular treatment of intracranial aneurysms using the Neuroform Atlas device at the Department of Neurology, People's Liberation Army General Hospital from July 2020 to May 2022. There were 34 males and 43 females, with a median (range) age of 59 (23-81) years. The degree and effect of aneurysm embolization were evaluated by modified Raymond grading post procedure and after 6 months. Complications occurring during the perioperative period were recorded. Vaso computed tomography was performed immediately after the operation to assess stent opening and adherence. Digital subtraction angiography was performed 6 months after discharge and the aneurysm was classified as cured, stable, or recurrent. Results: A total of 87 Atlases were successfully released in 77 cases. Angiography performed immediately after the embolization revealed 19 (24.7%) modified Raymond grade Ⅰ, 10 (13.0%) grade Ⅱ, and 48 (62.3%) grade Ⅲa cases. Three perioperative complications were observed including thrombotic events in 2 cases and stent migration in 1 case. A follow-up angiogram was available for 47 aneurysms, and showed that modified Raymond grade Ⅰ occlusion was achieved in 38 (80.9%) cases, grade Ⅱ in 2 (4.3%) cases, and grade Ⅲa in 7 (14.9%) cases. At the 6-month follow-up, 38 patients were cured and 7 were stable, whereas 2 patients experienced a recurrence of aneurysm. Stenosis of the parent artery occurred in 3 (6.4%) cases, including 2 at the head and 1 inside the stent. Conclusions: The results of this preliminary study suggest that Neuroform Atlas stent-assisted coiling has a high occlusion rate and low incidence of complications in the endovascular treatment of aneurysms. However, the effectiveness of this procedure for large aneurysms and long-term outcomes require further investigation.

Advances in the study of molecular identification technology of Echinococcus species.

The larvae of Echinococcus (hydatidcyst) can parasitize humans and animals, causing a serious zoonotic disease-echinococcosis. The life history of Echinococcus is complicated, and as the disease progresses slowly after infection, early diagnosis is difficult to establish. Due to the limitations of imaging and immunological diagnosis in this respect, domestic and foreign scholars have established a variety of molecular detection techniques for the pathogen Echinococcus over recent years, mainly including nested polymerase chain reaction (PCR), multiplex PCR, real-time quantitative PCR, and nucleic acid isothermal amplification technology. In this article, the research progress of molecular detection technology for Echinococcus infection currently was reviewed and the significance of these methods in the detection and diagnosis of hydatid and hydatid diseases was also discussed.

[Construction of zebrafish models for screening intracranial hemorrhage associated genes].

Objective: To construct zebrafish models for the screening of intracranial hemorrhage (ICH) associated genes. Methods: ICH zebrafish models were constructed through morpholino oligonucleotides (MOs) technique and microinjection technique, and multiple verification was performed from macro and micro perspectives. First, the normal wild-type AB strain zebrafish injected with control MO was used as the control group, and AB zebrafish embryos microinjected with MOs of genes related to development of neural crest-derived cells (NCDCs) were used as the study group, such as col8a1 MO, tfap2α MO, msx1a MO, msx2 MO, and dkk1a MO. Preliminary verification of the model was conducted under a white-light optical microscope. Then, the model was verified by Tg (flk1: gfp; gata1: dsRed) double transgenic zebrafish, with vascular endothelial cells labeled by green fluorescent protein (GFP) and red blood cell labeled by fluorescent protein (dsRed), and thus the location of cerebral hemorrhage can be observed more clearly. Specifically, zebrafish embryos were microinjected with Control MO as the control group and those microinjected with col8a1 MO as the study group. Then the embryos were cultured until 48 hours post-fertilization to observe the leakage of red blood cells under the confocal laser scanning microscope. Finally, Tg (flk1: gfp) transgenic zebrafish was used to verify the model based on the blood-brain barrier (BBB). Through the leakage of dextran-rhodamine and DAPI dyes, the destruction of BBB and the occurrence of cerebral hemorrhage in zebrafish were further clarified, and quantitative statistics were carried out to verify the relationship between NCDCs development related genes and cerebral hemorrhage phenotype, which proved that the modeling was effective. Results: The zebrafish with col8a1, tfap2α, and msx1 mutations in the study group had apparent ICH compared with wildtype zebrafish, and the prevalence of ICH was 18.18% (52/286), 23.04% (62/251), and 35.94% (23/64), respectively. While, the zebrafish with msx2 and dkk1a mutations rarely had ICH, with the ICH prevalence of 1.03% (1/97) and 1.15% (1/87), respectively. The prevalence of red blood cells leakage in Tg (flk1:gfp; gata1:dsred) double transgenic zebrafish injected with Control Mo and col8a1 Mo was 0.37% (1/273) and 18.18% (52/286) (P<0.001). The number of DAPI positive nuclei of Tg (flk1: gfp) transgenic zebrafish injected with Control Mo and col8a1 Mo was 10.05±5.27 and 60.35±3.96 (P<0.001), and the fluorescent intensity of midbrain parenchymal induced by dextran-rhodamin leakage was 2.54±4.70 and 5.13±3.52 (P<0.001). Conclusion: This study successfully constructs the ICH zebrafish models, and ICH-related genes are screened out, such as col8a1, tfap2α, msx1, and so on.

[Effect of D-dimer on the prognosis of patients with aneurysmal subarachnoid hemorrhage based on propensity score matching].

Objective: To evaluate the effect of D-dimer on the prognosis of patients with aneurysmal subarachnoid hemorrhage (aSAH). Methods: A total of 1 658 patients who were first diagnosed with aSAH in West China Hospital of Sichuan University from December 2013 to June 2019 were retrospectively analyzed. All patients were divided into four groups according to the median and quartiles of D-dimer level, including 415 cases, 414 cases, 414 cases, and 415 cases in groups Q1, Q2, Q3, and Q4, respectively. Groups Q2, Q3, Q4, and group Q1 were matched by propensity score matching (PSM), and the correlation between D-dimer and each outcome was analyzed by logistic regression. Since there is no general clinical classification standard for D-dimer, this study attempted to reclassify patients into groups q1 (<0.55 mg/L, 94 cases), q2 (0.55-1.65 mg/L, 435 cases), q3 (1.65-5.50 mg/L, 650 cases) and q4 (>5.50 mg/L, 303 cases) based on 1, 3, 5, 10 times of the upper limit of the current clinical reference value. Results: The age of 1 658 aSAH patients were (57±12) years, including 1 068 males and 590 females. After PSM based on the median and quartiles of D-dimer level, there were 318 cases, 318 cases, 251 cases, and 229 cases in groups Q1, Q2, Q3, and Q4, respectively. Compared with group Q1 (<1.23 mg/L), the risk of in-hospital infection (OR=2.14, 95%CI: 1.47-3.11, P<0.001), pneumonia (OR=2.22, 95%CI: 1.51-3.28, P<0.001), urinary tract infection (OR=1.75, 95%CI: 1.12-2.75, P=0.014) and intracranial rebleeding (OR=3.59, 95%CI: 1.30-9.91, P=0.013) group Q4 (>4.95 mg/L) was higher. Likewise, the risk of adverse outcomes in group Q4 was also higher than that in group Q1, including unfavorable outcome at discharge (OR=2.12, 95%CI: 1.43-3.14, P<0.001), mortality during hospitalization (OR=3.03, 95%CI: 1.26-7.33, P=0.014), mortality within 90 days (OR=2.33, 95%CI:1.29-4.22, P=0.005), mortality within 180 days (OR=1.92, 95%CI: 1.12-3.29, P=0.018), mortality within 1 year (OR=2.07, 95%CI:1.23-3.47, P=0.006) and mortality during the longest follow-up period (OR=1.97, 95%CI:1.26-3.09, P=0.003). After secondary grouping and PSM based on current clinical reference values, there were 90 cases, 87 cases, 90 cases, and 43 cases, respectively in groups q1, q2, q3 and q4. The risk of nosocomial infection (OR=2.26, 95%CI: 1.14-4.45, P=0.019), blood-borne infection (OR=8.86, 95%CI:1.08-72.78, P=0.042), poor prognosis at discharge (OR=4.92, 95%CI: 2.18-11.07, P<0.001), death within 180 days (OR=3.39, 95%CI: 1.04-11.08, P=0.043), death within 1 year (OR=3.23, 95%CI: 1.10-9.49, P=0.033), and death within the longest follow-up period (OR=3.28, 95%CI: 1.34-8.01, P=0.009) was still higher in group q4 than that in group q1. Conclusion: aSAH patients with high D-dimer level have a higher risk of complications and mortality during hospitalization and worse clinical prognosis.

[Clinical analysis of pulmonary nocardiosis associated with bronchiectasis].

Objective: To better understand the clinical characteristics of pulmonary nocardiosis associated with bronchiectasis. Methods: Patients diagnosed as bronchiectasis complicated with pulmonary nocardiosis in 9 tertiary general hospitals in China were enrolled from March 2016 to March 2020, with the record of general data, imaging performance and pathogen. The literature was reviewed. Results: Totally 17 patients were included. There were 12 females and 5 males. The ages ranged from 45 to 79 years, with an average of (63±9) years. There were 15 nonsmokers and 2 smokers, all of whom with chronic course. The clinical manifestations were mostly cough, expectoration, hemoptysis, fever, and dyspnea. The imaging manifestation was bronchiectasis in both lungs, with the most common involvement in the left lower lung, right middle lobe and left lingual lobe. Sputum cultures were positive in 10 cases, bronchoalveolar lavage fluid (BALF) cultures were positive in 6 cases, and next generation gene sequencings were positive in 4 cases, including 2 cases of Nocardia gelsenkii, 2 cases of Nocardia abscess, 2 cases of Nocardia stellate, 1 case of Nocardia mexicana, 1 case of Nocardia otitis caviae, and 9 cases of undetermined Nocardia. There were 3 cases of Klebsiella pneumoniae, 2 cases of Pseudomonas aeruginosa and 2 cases of Aspergillus. The symptoms and imaging of all patients were improved after anti Nocardia therapy. Conclusions: Bronchiectasis combined with nocardiosis is more common in middle-aged and elderly women without smoking, which is similar to the clinical manifestations of Lady Windermere syndrome. Bronchiectasis often involves the left lower lobe, right middle lobe and left lingual lobe. Nocardia infection might further precipitate the initiation and progression of bronchiectasis.

[Mechanism of hepatic fibrosis associated with Echinococcus: a review].

Echinococcosis is a zoonotic parasitic disease caused by Echinococcus infections, and this disorder may cause fibrosis of multiple vital organs, which may further progress into cirrhosis. Early-stage hepatic fibrosis is reversible, and unraveling the mechanisms underlying hepatic fibrosis induced by Echinococcus infections is of great significance for the prevention and treatment of early-stage hepatic fibrosis. Recently, the studies pertaining to hepatic fibrosis associated with Echinococcus infections focus on cytokines and immune cells. This review summarizes the advances in the mechanisms underlying host immune cells- and cytokines-mediated hepatic fibrosis in humans or mice following Echinococcus infections.

[Application expansion and related problems of lumbar cortical bone trajectory screw technique].

At the beginning of clinical application, CBT technique was mainly used for lumbar degenerative diseases in Elderly Patients, and single or double segments of lower lumbar vertebrae accounted for the majority. With the technical proficiency and equipment improvement, the types of diseases continue to expand, and CBT technique can be used in conjunction with traditional pedicle screw technology, which plays an important role in the revision of adjacent segment disease and fixation in lumbar vertebrae of patients with severe osteoporosis. However, CBT technology has not been widely used, and there are still related problems that need to be clarified and further studied.

[Pathogen distribution and antimicrobial resistance among lower respiratory tract infections in patients with hematological malignancies].

Objective: To investigate the pathogen distribution and antimicrobial resistance among lower respiratory tract infections in patients with hematological malignancies. Methods: Sputum samples were collected from 967 patients with hematological malignancies and lower respiratory tract infections in Department of Hematology,the Second Hospital of Shanxi Medical University from January 2017 to July 2020. The pathogens and drug sensitivity reports were carried out by automatic bacterial identification instruments. WHONET 5.6 and SPSS 20.0 softwares were used for statistical analysis. Results: A total of 961 strains of pathogens were isolated, 516 (53.7%) pathogens were Gram-negative bacteria, mainly 118 strains of Klebsiella pneumonia (12.3%), 68 strains of Pseudomonas aeruginosa (7.1%), 67 strains of Acinetobacter baumannii (7.0%),52 strains of Stenotrophomonas maltophilia (5.4%), 43 strains of Escherichia coli (4.5%), and 42 strains of Enterbacter cloacae (4.4%). There were 171 (17.8%) strains of Gram-positive bacteria and 274 (28.5%) fungi. The drug resistance rates of Pseudomonas aeruginosa and Acinetobacter baumannii to carbapenem were 22.1%-31.3%. Stenotrophomonas maltophilia was sensitive to levofloxacin, compound sulfamethoxazole and minocycline. The antimicrobial resistance rates of these three enterobacteria to carbapenems, cefoperazone/sulbactam, piperacillin/tazobactam were low (<10%). The resistant Gram-positive bacteria to ticoplanin, vancomycin and linazolamide were not detected. Conclusion: The major pathogens related to lower respiratory tract infections in patients with hematological malignancies are gram-negative bacteria in our centre. Different pathogens appear different characteristics of antimicrobial resistance.

[Transcription factor EB related autophagy in the treatment of multiple myeloma and its mechanism].

Objective: To clarify the effects of bortezomib combined with or without siramesine on the proliferation of multiple myeloma cell lines, the expression changes of transcription factor EBC (TFEB) nuclear translocation and the level of autophagy, and to provide basis for further exploring the regulation mechanism of transcription factor TFEB on autophagy. Methods: The multiple myeloma cell lines RPMI8226 and U266 were cultured in vitro, and the multiple myeloma cells were treated with a certain concentration of bortezomib and siramesine. The changes of cell proliferation inhibition were detected by CCK-8 method. Real time PCR and Western blot were used to detect the relative expression of TFEB, autophagy-related factor LC3B, Beclin1, p62, LAMP1 mRNA and protein. Results: As the concentration of bortezomib increased and the duration of action increased, the proliferation inhibition rates of the two cell lines gradually increased (P<0.05) . The combination of the two drugs has a synergistic inhibitory effect on the proliferation of the above-mentioned multiple myeloma cell lines (P<0.05) . In the blank control group, single drug group, and combination drug group, the relative expression of TFEB mRNA and protein in the cytoplasm decreased sequentially (P<0.05) , and the relative expression of TFEB mRNA and protein in the nucleus increased sequentially (P<0.05) . The relative expression of autophagy-related factors LC3B, Beclin1, LAMP1 mRNA and protein increased sequentially, and the relative expression of p62 mRNA and protein decreased sequentially (P<0.05) . Conclusion: Bortezomib and siramesine can synergistically inhibit the growth of multiple myeloma cells, which is related to the increased autophagy expression in multiple myeloma cell lines and the expression of TFEB with nuclear translocation is also enhanced.

[Endovascular stenting for idiopathic intracranial hypertension with different types of venous sinus stenosis].

Objective: To evaluate the efficacy of endovascular stenting of various types of venous sinus stenosis in idiopathic intracranial hypertension (IIH). Method: Clinical, radiological, and manometric data before and after stenting in venous sinus stenosis were retrospectively analyzed in 99 IIH patients who were refractory to medical therapy or rapidly progressed between July 2004 to July 2019. The follow-up period was between 2.3 months to 11 years. Results: Our study enrolled 21 men (21.2%)and 78 women (78.8%) with average body mass index (BMI) 19.2-40.6(27.0±4.4) kg/m2 and median age 37 years. Before stent placement, the mean transverse sinus stenosis gradient was 1-59(26±8) mmHg. Patients with extrinsic stenosis were younger than those with intrinsic and mixed stenosis. In all cases, stenting was effective for papilledema. Fifty patients complained of headaches. Pulsatile tinnitus in twenty-eight patients completely alleviated after stenting. In one patient, replacement of stent did not improve symptoms, and a subsequent CSF diversion procedure was performed and effective. Conclusion: Irrespective of the type of stenosis, stenting of venous sinus stenosis is an effective treatment for IIH. Patients with persistent papilledema post-stenting and elevated transverse pressure pre-stenting should be followed closely as high risk of stenting failure may occur and further diversion procedure is needed.

Hypomethylating agents for elderly patients with acute myeloid leukemia: a PRISMA systematic review and meta-analysis.

OBJECTIVE: Abnormal DNA methylation plays a critical role in acute myeloid leukemia (AML) pathogenesis and hypomethylating agents (HMAs) such as decitabine (5-aza-29-deoxycytidine) and azacitidine (5-azacytidine) are considered efficacious for treating AML. This study aimed to identify if HMAs have therapeutic advantages compared with conventional care regimens (CCR) or placebo in elderly AML patients. MATERIALS AND METHODS: We systematically searched PubMed, Embase, and Cochrane Central Register of Controlled Trials from inception to November July 15, 2020. Randomized controlled trials that compared the efficacy and adverse events associated with HMAs, CCR, or placebo were searched. RevMan 5.3 software was used to calculate the hazard ratio (HR) and risk ratio (RR) with a 95% confidence interval (CI). RESULTS: Seven trials with a total of 1966 participants were included. Meta-analyses showed that the overall survival of HMAs was better than that of CCR [HR=0.76, 95% CI (0.69-0.85), (p<0.01)], and the complete remission rate of elderly AML patients was increased by HMAs compared with CCR [RR=1.46, 95%CI (1.08-1.99), p=0.01)]. HMA treatment showed higher incidence of neutropenia [RR=1.30 (95%CI 1.07-1.59, p=0.008)], thrombocytopenia [RR=1.14 (95%CI 1.01-1.59, p=0.04)], and pneumonia [RR=1.37 (95%CI 1.06-1.76, p=0.02)] compared with CCR. CONCLUSIONS: Although HMAs cause a higher incidence of adverse events such as neutropenia, thrombocytopenia, and pneumonia, demethylation drugs are well-tolerated and effective for treating AML in the elderly.

[Preliminary study on the effect of vascular endothelial growth factor-loaded self-assembled peptide hydrogel on angiogenesis and vascularization of human umbilical vein endothelial cells].

Objective: To investigate the effect of RATEA16 scaffold on the proliferation of human umbilical vein endothelial cells (HUVEC) and the effect of new self-assembling peptide hydrogel (RATEA16) scaffold with vascular endothelial growth factor (VEGF) on promoting angiogenesis. Methods: RATEA16 hydrogel was prepared, then the injectability, microstructure, degradation, biocompatibility of RATEA16 hydrogel were determined. HUVEC were cultured with RATEA16 scaffold to detect cell morphology and proliferation. HUVEC were cultured on RATEA16 scaffold with VEGF for 24 h. The expression of VEGF-A, von Willebrand factor (vWF), matrix metalloproteinase-9 (MMP-9) and platelet endothelial cell adhesion molecule-1 (PECAM-1) were detected by using real-time PCR to evaluate the effects of the scaffold with VEGF system on HUVEC differentiation. Results: The sol-gel transition was completed under neutral condition (pH=7.4) adjusted by Tris-HCl solution. The hydrogel could be easily injected from a syringe. It presented a porous and interconnected internal structure and the porosity of the scaffold was (67.3±9.4)%. After 4 week degradation in vitro, the residual weight was still (82.354±0.006)%, which exhibited slow degradation. HUVEC grew well after being cultured in leach liquor of RATEA16 hydrogel for 24 h, and there was no significant difference in HUVEC cell viability compared with that of the control group (P>0.05). HUVEC encapsulated in RATEA16 hydrogel appeared round in shape and exhibited effectively continuous proliferation. When HUVEC were cultured on RATEA16 hydrogel with VEGF for 24 h, the formation of vascular-like structures was observed. The expression of VEGF-A and MMP-9 was 1.5-2.0 times that of control group, and vWF was 10 times and PECAM-1 was 55 times compared with that of the control group (P<0.05). Conclusions: The RATEA16 hydrogel used in this study could be prepared by simply adjusting pH to neutral. This hydrogel exhibited good biodegradability, slow degradation and injectability. HUVEC might attach and spread in RATEA16 scaffold. The RATEA16 scaffold with VEGF could promote angiogenic differentiation of HUVEC. The novel scaffold is expected to achieve the critical vascularization process in bone tissue regeneration.

[Feasibility of the determination of plasma vardenafil level in rat by performance liquid chromatography-tandem mass spectrometry].

Objective: To develope and validate a reliable and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of vardenafil concentration in plasma of rat. Methods: Plasma samples of normal Sprague-Dawley rats were collected. A Phenomenex Synergi Polar-RP 80A column (2.0 mm×50 mm, 4 µm) was used. Column temperature was set at 30 ℃. Mobile phase A was 0.1% formic acid in water; mobile phase B was 0.1% formic acid in acetonitrile. The flow rate was 0.4 ml/minutes. Quantitative determination was performed by electrospray ionization, operating in positive ion multiple reaction monitoring (MRM) mode. Cisapride was used as the internal standard. The feasibility of the method was evaluated by examining its specificity, linearity and quantitative range, precision and accuracy, matrix effects, and stability. Results: Under the selected chromatographic and mass spectrometry conditions, the monitoring ions of vardenafil and internal standard were mass-to-charge ratio（m/z) 489.3/151.2 and 466.4/234.2, the retention times of vardenafil and internal standard were 2.62 and 2.80 minutes, respectively, and the peak shape was satisfactory. The method has good linearity in the concentration range of 0.2-200 ng/ml. The intra-batch precision (%CV) and accuracy (%DEV) of vardenafil were 1.5%-9.7% and -6.8%-6.6%, respectively. The inter-batch precision and accuracy of vardenafil were 3.1% -8.4% and -3.7%-4.6%, respectively. In this sample processing method, the extraction recovery rate of vardenafil was obtained at range of 88.2%-104.6%, which met the requirements for the investigation of extraction recovery rate. In this sample processing method, the normalized matrix factor of each quality control concentration of vardenafil was 1.04, 0.85, and 1.04, and the coefficient of variation (%CV) was in the range of 1.7%-10.7%, which met the requirements for the investigation of matrix effects. Variations of short-term stability, long-term stability, and stability of 4 freeze-thaw cycles of vardenafil was within ±15%, and the coefficient of variation were within 5%. Conclusion: The high performance liquid chromatography-tandem mass spectrometry method established in this study is feasible for the measurement of concentration of vardenafil in rat plasma and this method has good specificity and high accuracy, and can be used to detect the concentration of vardenafil in rat plasma.

[Relationship between mutations of HBV basal core promoter region in HBsAg-positive mothers and intrauterine transmission].

Objective: To analyze the relationship between maternal mutations in basal core promoter region of hepatitis B virus (HBV) genotype C and intrauterine transmission. Methods: We collected information on general demographic characteristics and process of delivery among 399 pairs of consecutive HBsAg-positive mothers and their neonates, from the Third People's Hospital of Taiyuan in Shanxi province, China. Fluorescence quantitative polymerase chain reaction (FQ-PCR) and Electro-chemiluminescence immuno-assay (ECLIA) kits were used to detect both maternal and neonatal HBV DNA and serological markers in the peripheral blood. From 113 mothers with HBV DNA load ≥10(6) IU/ml, we selected 22 mothers whose neonates were with intrauterine transmission and randomly selected the same number of mothers whose neonates were without intrauterine transmission, as controls. The whole-length HBV DNA were extracted, amplified, cloned, sequenced and genotyped. Finally, a total of 39 mothers with genotype C of HBV were selected for mutation analysis. Results: Thirty-nine cases of genotype C (88.63%) were finally included in the study, with 19 cases in the intrauterine transmission group and 20 cases as controls. Rates of A1762T/G1764A double mutations were significantly different between the intrauterine transmission group and the control group (7.53% vs. 27.72%, P<0.001). Results from the multivariate analysis showed that the A1762T/G1764A double mutations had reduced the risk of intrauterine transmission (aOR=0.065, 95%CI: 0.006-0.746, P=0.028). Maternal A1762T/G1764A double mutations appeared to be possibly associated with neonatal HBeAg (P=0.050). Conclusion: A1762T/G1764A double mutations of HBV DNA from the genotype C of those HBsAg-positive mothers could reduced the risk of HBV intrauterine transmission during pregnancy.