Clinical and genetic characteristics analysis of four cases of Phelan-McDermid syndrome caused by the novel mutations of the SHANK3 gene / 中华实用儿科临床杂志

The basic information and clinical data of 4 Phelan-McDermid syndrome (PMS) patients in the Pediatric Outpatient Department of the Peking University First Hospital from January 2014 to October 2019 were retrospectively analyzed.Genetic diagnoses were performed using the whole exon sequencing assay.The genotype-phenotype correlation analysis was then performed.All patients presented with intellectual disability/developmental delay, especially the most-common manifestation in language disability.Patient 2 had an autism behavior.Four novel variations of the SHANK3 gene were found in this study, including the c. 2861delC p. (S955Pfs*109), c.3166delC p. (A1039Afs*39), c.3711_3723delGCCCAGCCCCCGG p. (L1241Lfs*29) and c. 2223+ 1G>A.All of them were analyzed as new pathogenic variations according to the American College of Medical Genetics and Genomics criteria.The present study expan-ded the mutant spectrum of the SHANK3 gene, which provided a basis for further accurate genetic counseling and prenatal diagnosis of PMS.

Effect of Yinxing Mihuan Oral Solutionon P2RX7/NLRP3 Signaling Pathway in Depression Model Rats Induced by Isolation Combined with Chronic Unpredictable Mild Stress / 中国实验方剂学杂志

Objective:To explore the antidepressant mechanism of Yinxing Mihuan oral solution （YMO） by investigating its effect on depression model rats. Method:The depression rats were induced by isolation combined with chronic unpredictable mild stress （CUMS） and then randomly divided into model group， fluoxetine group （10 mg·kg^-1） and high-dose （618 mg·kg^-1） and low-dose （309 mg·kg^-1） YMO groups. A blank control group was also set up and ten rats were included in each group. Modeling lasted for 21 consecutive days， and rats were administered the 8th day after stimulation at a dose of 10 mL·kg^-1 for 14 days， except those in the blank control and model groups which were given distilled water. Afterward， the sucrose preference test， open field test， tail suspension test were carried out. The pathological changes of hippocampus in depression rats were observed after hematoxylin-eosin （HE） staining. The content of interleukin-1<italic>β </italic>（IL-1<italic>β</italic>）， interleukin-6 （IL-6） and tumor necrosis factor-<italic>α </italic>（TNF-<italic>α</italic>） in the hippocampus of rats in each group and the expression of NOD-like receptor 3 （NLRP3） and other proteins in its related activation signaling pathways were detected with multi-factor detection （Luminex） and Western blot. Result:After 14 days of continuous administration， compared with the blank control group， the model group witnessed significantly reduced sugar water consumption rate and the times of rearing and significantly prolonged cumulative time of immobility during tail suspension （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the model group， the fluoxetine group and the high-dose YMO group saw increases in the times of rearing， times of crossing and sugar water consumption rate and a significant decrease in the cumulative time of immobility during tail suspension （<italic>P</italic><0.05， <italic>P</italic><0.01）. The results of HE staining showed that the neurons in the hippocampus of rats in the high-dose YMO group were arranged in order and slightly loosened， without obvious microglia infiltration observed. The levels of IL-1<italic>β</italic>， IL-6 and TNF-<italic>α</italic> in the hippocampus of the model group increased significantly as compared with the blank control group （<italic>P</italic><0.05， <italic>P</italic><0.01）， and their content in the high-dose YMO group was significantly lowered in the comparison with the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Molecular biology experiments demonstrated that compared with the results of blank group， the expression of purinergic receptor P2X7 （P2RX7）， NLRP3， apoptosis-associated speck-like protein （ASC）， Caspase-1 and IL-1<italic>β</italic> remarkably increased in the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Additionally， the expression of P2RX7， NLRP3， ASC， Caspase-1 and IL-1<italic>β </italic>was significantly inhibited in the fluoxetine group and the high-dose YMO group compared with the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:YMO can improve the depression-like behaviors of rats induced by isolation combined with CUMS， and its mechanism of action is related to the regulation of the P2RX7/NLRP3 signaling pathway.

Effects of calcium-sensitive receptors on 11β-hydroxysteroid dehydrogenase type 2 and cortisol in neonatal mice with persistent pulmonary hypertension / 中国当代儿科杂志

OBJECTIVE@#To investigate the effects of calcium-sensitive receptors (CaSR) on the expression of 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) and cortisol concentration in a neonatal mouse model of persistent pulmonary hypertension (PPH).@*METHODS@#Fifty-six newborn C57BL/6 mice were randomly divided into a control group (n=14), a PPH group (n=14), an agonist group (n=14), and an inhibitor group (n=14). The mice in the PPH, agonist, and inhibitor groups were exposed to a 12% oxygen concentration, and the agonist group and inhibitor group were given CaSR agonist (GdCl3, 16 mg/kg) and CaSR antagonist (NPS2390, 1 mg/kg) intraperitoneally once a day, respectively. The mice in control group were exposed to air, and then injected with an equal volume of normal saline as those in the PPH group every day. All mice were treated for 14 days. Morphological examination of heart and lung tissues was performed using hematoxylin-eosin staining. The expression levels of 11β-HSD2 mRNA and 11β-HSD2 protein in lung tissues were measured by qRT-PCR and Western blot respectively. Brain natriuretic peptide (BNP) and cortisol levels in lung tissues were determined using ELISA.@*RESULTS@#Compared with the control group, the PPH group had significantly increased pulmonary artery wall thickness (WT%), ratio of right to left ventricular thickness (RV/LV), alveolar mean linear intercept, and BNP concentration and a significantly reduced radial alveolar count (P<0.05); compared with the PPH group, the agonist group showed significant increases in WT% and BNP concentration, while the inhibitor group showed significant reductions in the two indicators (P<0.05). Compared with the control group, the PPH group showed significant reductions in the expression levels of 11β-HSD2 mRNA and 11β-HSD2 protein, but a significant increase in cortisol concentration (P<0.05); compared with the PPH group, the agonist group had significantly lower expression levels of 11β-HSD2 mRNA and 11β-HSD2 protein, but a significant higher cortisol concentration, while the inhibitor group showed opposite changes in these indicators (P<0.05).@*CONCLUSIONS@#CaSR may control the development and progression of PPH in newborn mice by regulating the expression of 11β-HSD2 and cortisol concentration.

Effect of calcium-sensitive receptors on endothelial nitric oxide synthase and nitric oxide in neonatal mice with persistent pulmonary hypertension / 中国当代儿科杂志

OBJECTIVE@#To study the effect of calcium-sensitive receptors (CaSR) on the expression of endothelial nitric oxide synthase (eNOS) and the concentration of nitric oxide (NO) in a neonatal mouse model of persistent pulmonary hypertension (PPH).@*METHODS@#Eighty neonatal C57BL/6 mice were randomly divided into control, PPH, agonist and antagonist groups. The control group was exposed to air, and the other three groups were exposed to 12% oxygen. The agonist and antagonist groups were intraperitoneally injected with a CaSR agonist (GdCl 16 mg/kg) and a CaSR antagonist (NPS2390, 1 mg/kg), respectively, while the PPH and control groups were intraperitoneally injected with normal saline instead. All mice were treated for 14 days. Alveolar development and pulmonary vessels were assessed by hematoxylin-eosin staining. The protein and mRNA expression of eNOS and its localization in lung tissues were determined by Western blot, qRT-PCR and immunohistochemistry. The levels of brain natriuretic peptide (BNP) and NO in lung homogenate were determined using ELISA.@*RESULTS@#Compared with the control group, the PPH and agonist groups showed significant increases in alveolar mean linear intercept, the percent wall thickness of pulmonary arterioles, right to left ventricular wall thickness ratio (RV/LV) and BNP concentration, but a significant reduction in radial alveolar count (P<0.05). The antagonist group had significant improvements in all the above indices except RV/LV compared with the PPH and agonist groups (P<0.05). Compared with those in the control group, the protein and mRNA expression of eNOS and NO concentration were significantly increased in the PPH group and increased more significantly in the agonist group, but were significantly reduced in the antagonist group (P<0.05).@*CONCLUSIONS@#CaSR plays an important role in the development of PPH in neonatal mice, possibly by increasing eNOS expression and NO concentration.

Mindfulness Meditation for Primary Headache Pain: A Meta-Analysis / 中华医学杂志(英文版)

<p><b>Background</b>Several studies have reported that mindfulness meditation has a potential effect in controlling headaches, such as migraine and tension-type headache; however, its role remains controversial. This review assessed the evidence regarding the effects of mindfulness meditation for primary headache pain.</p><p><b>Methods</b>Only English databases (PubMed, Cochrane Central Register of Controlled Trials [the Cochrane Library], PsycINFO, Psychology and behavioral science collection, PsyArticles, Web of Science, and Scopus) were searched from their inception to November 2016 with the keywords ("meditation" or "mindfulness" or "vipassana" or "dzogchen" or "zen" or "integrative body-mind training" or "IBMT" or "mindfulness-based stress reduction" or "MBSR" or "mindfulness-based cognitive therapy" or "MBCT" and "Headache" or "Head pain" or "Cephalodynia" or "Cephalalgia" or "Hemicrania" or "Migraine"). Titles, abstracts, and full-text articles were screened against study inclusion criteria: controlled trials of structured meditation programs for adult patients with primary headache pain. The quality of studies included in the meta-analysis was assessed with the Yates Quality Rating Scale. The meta-analysis was conducted with Revman 5.3.</p><p><b>Results</b>Ten randomized controlled trials and one controlled clinical trial with a combined study population of 315 patients were included in the study. When compared to control group data, mindfulness meditation induced significant improvement in pain intensity (standardized mean difference, -0.89; 95% confidence interval, -1.63 to -0.15; P = 0.02) and headache frequency (-0.67; -1.24 to -0.10; P = 0.02). In a subgroup analysis of different meditation forms, mindfulness-based stress reduction displayed a significant positive influence on pain intensity (P < 0.000). Moreover, 8-week intervention had a significant positive effect (P < 0.000).</p><p><b>Conclusions</b>Mindfulness meditation may reduce pain intensity and is a promising treatment option for patients. Clinicians may consider mindfulness meditation as a viable complementary and alternative medical option for primary headache.</p>

Laparoscopy and short poly FURS for treatment of ureteropelvic junction obstruction combined with renal stones / 中国内镜杂志

Objective To evaluate the clinical efficacy of laparoscopy and short poly FURS in treatment of UPJO combined with renal stones. Methods From May 2014 to November 2016, 20 patients with ureteropelvic junction stricture in our hospital were treated with Laparoscopy and short poly FURS lithotripsy, the clinical data of the patients before and after operation were retrospectively analyzed. Results The patients in this group were successfully operated in stage I, and the hemoglobin descent rate did not change significantly before and after operation. No need of blood transfusion, the first day after the review of urinary tract plain film observed DJ tube position, postoperative back to hospital in March pulled out of DJ tube, postoperative follow-up 6 months to 1 year, the patient recovered well, no long-term complications. Conclusion Laparoscopy and short poly FURS lithotripsy, has satisfactory effects and no significant complications which should be promoted.

Effect of calcium-sensitive receptor on intracellular Ca2+ concentration in pulmonary arterial smooth muscle cells of hypoxia-induced persistent pulmonary hypertension in mice / 中国新生儿科杂志

Objective To study the changes of intracellular calcium ion concentration in pulmonary artery smooth muscle cells (PASMCs) of hypoxic-induced persistent pulmonary hypertension (PPH) induced by calcium-sensitive receptor (CaSR) in a newborn mouse model.Method Ninety-six newbom C57BL/6 mice were randomly divided into control group,PPH group,PPH + agonist group and PPH + inhibitor group,with 24 mice in each group.The PPH model was induced by 12％ oxygen for 14 days.In the beginning,intraperitoneal injection of CaSR agonist (GdCl3) and CaSR inhibitor (NPS2390) were performed to mice in PPH + agonist group and PPH + inhibitor group respectively daily.After 14 days of modeling,pulmonary artery smooth muscle cells (PASMCs) of all four groups were cultured in vitro.Changes of Ca2+ fluorescence intensity in PASMCs of the four groups were detected by laser confocal microscope continuously.Result The ratio of pulmonary small vascular wall thickness to the vascular diameter and right ventricle/left ventricular thickness in PPH group were greater than those in the control group [(21.1％ ±1.8％) vs.(27.0％ ±0.9％),(0.62 ±0.22) vs.(0.83±0.45)],the differences were statistically significant (P ＜ 0.05),which imply that PPH mouse model was constructed successfully.The average Ca2+ fluorescence intensity in PASMCs of control group,PPH group,PPH + agonist group and PPH+ antagonist group were 122.5 ± 3.0,2 058.8 ±46.3,2 286.6 ±51.4 and 1 134.8 ± 8.5,respectively.The average Ca2+ fluorescence intensity in PASMCs of the PPH group,PPH + agonist group and PPH + antagonist group was higher than that of the control group respectively,the average Ca2+ fluorescence intensity in PASMCs of PPH group was higher than that of PPH + antagonist group,the differences were statistically significant (P ＜ 0.05).Whereas the difference of average Ca2 + fluorescence intensity in PASMCs of PPH group and PPH + agonist group was of no statistical significance (P ＞ 0.05).Conclusion CaSR may be involved in the occurrence and development of hypoxic-induced PPH in neonatal mice by affecting the intracellular Ca2+ concentration in pulmonary artery smooth muscle cells.

Simultaneous determination and pharmacokinetics of eight ginsenosides by LC-MS/MS after intravenously infusion of 'SHENMAI' injection in dogs

SHENMAI injection, a prescription comprised of Panax ginseng and Ophiopogon japonicas, is being extensively applied in the field of cardio-protection and immune-modulation in China. Ginsenosides are the main active components in SHENMAI injection. In order to capture and analyze the pharmacokinetic profile of major ginsenosides of SHENMAI injection in Beagle dogs, liquid chromatography equipped with electro-spray ionization and tandem mass spectrometry method was applied in simultaneous determination for protopanaxatriol type ginsenoside [Re, Rf, Rg1], protopanaxadiol type ginsenoside [Rb2, Rb1, Rd, Rc] and oleanolic acid type ginsenoside [Ro]. A C18 column [150 × 2.1mm, 5micro m] and a linear gradient program were used to achieve chromatographic separation, with 0.02% acetic acid solution and acetonitrile. I.S. and ginsenosides were detected by LC-MS/MS in selective reaction mode. Good linearity spanning 5- 1500ng/mL was achieved with the R[2] values higher than 0.99 for all analytes. Limit of quantification of all analytes were 3ng/mL. Intra- and inter-day precisions ranges from 0.47 to15.68 % and accuracies were within the range of 85.27-117.57%. Validated analyzing method was then used in the pharmacokinetic experiment for SMI in dogs. The results showed that the pharmacokinetic profile of protopanaxadiol, protopanaxatriol and oleanolic acid type ginsenoside were significant difference in dogs. Protopanaxadiol type ginsenosides exhibited an extremely higher level of exposure and a much slower elimination process. Whereas protopanaxatriol type ginsenosides were quickly eliminated. We concluded that 20 [S] - protopanaxadiol type ginseno sides could be a potential pharmacokinetic marker of SHENMAI injection

Role of calcium-sensing receptor in neonatal mice with persistent pulmonary hypertension / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of calcium-sensing receptor (CaSR) agonists and antagonists on the expression of CaSR in neonatal mice with persistent pulmonary hypertension (PPHN), and to clarify the role of CaSR in neonatal mice with PPHN.</p><p><b>METHODS</b>Forty-nine neonatal mice were randomly divided into four groups: control (n=10), hypoxia (PPHN; n=11), agonist (n=13), and antagonist (n=15). The mice in the PPHN, agonist, and antagonist groups were exposed to an oxygen concentration of 12%, and those in the control group were exposed to the air. The mice in the agonist and antagonist groups were intraperitoneally injected with gadolinium chloride (16 mg/kg) and NPS2390 (1 mg/kg) respectively once daily. Those in the PPHN and the control groups were given normal saline daily. All the mice were treated for 14 consecutive days. Hematoxylin and eosin staining and immunohistochemistry were used to observe the changes in pulmonary vessels. Laser confocal microscopy was used to observe the site of CaSR expression and measure its content in lung tissues. qRT-PCR and Western blot were used to measure the mRNA and protein expression of CaSR in lung tissues.</p><p><b>RESULTS</b>Compared with the control group, the PPHN group had significant increases in the pulmonary small artery wall thickness and the ratio of right to left ventricular wall thickness (P<0.05), which suggested that the model was successfully prepared. Compared with the control group, the PPHN group had a significant increase in the mRNA and protein expression of CaSR (P<0.05), and the agonist group had a significantly greater increase (P<0.05); the antagonist group had a significant reduction in the mRNA and protein expression of CaSR (P<0.05).</p><p><b>CONCLUSIONS</b>CaSR may play an important role in the development of PPHN induced by hypoxia in neonatal mice.</p>

MRI features of phenylketonuria in brain / 中国医学影像技术

Objective To observe MRI features of intracranial changes in phenylketonuria (PKU) patients.Methods Brain MRI findings of clinically diagnosed 13 PKU patients were analyzed retrospectively.The structure and signal changes of cerebrum,cerebellum and brainstem were observed.Results Posterior horn and triangle high signals were shown in white matter of bilateral lateral ventricle of all 13 patients on T2WI,while the optic radiation was spared.Periventricular white matter abnormal signals at body and anterior horn bilateral lateral ventricle (n=12),bilateral basal ganglia involvement (n=1),extensive white matter involvement (n=1),backward myelination and agenesis of corpus callosum (n=3),and mild macrogyria deformity (n=1) were found.The high signals in lateral periventricular white matter were shown on DWI,with reduction of ADC values in 6 cases.Conclusion Abnormal high signals in brain posterior horn and triangle white matter,optic radiation spared,and corpus callosum and cortical malformations are brain MRI appearances in brain of PKU.

Cell suspension and level of IL-17 in bronchoalveolar lavage fluid (BALF) of children with labor pneumonia combined with mycoplasma pneumonia and its clinical significance / 中国内镜杂志

Objective To explore cell suspension and level of IL-17 in bronchoalveolar lavage fluid (BALF) of children with labor pneumonia combined with mycoplasma pneumonia and its clinical significance. Methods 87 children with labor pneumonia who had been done BAL were divided into CPIS ≥ 6 group and CPIS 0.05). The percentage of columnar epithelial cells in CPIS < 6 group were higher than CPIS ≥ 6, how-ever, macrophagocyte was lower ( < 0.05). The percentage of neutrophils are related with both MP-DNA ( = 0.48,= 0.000) and IL-17 ( =0.49, =0.000). Conclusions The composition of BALF is more appropriate to react the circumstance of lobar pneumonia. When lobar pneumonia occurs, columnar epithelial cells are damaged, macrophagocyte gathered to participate in the inflammatory response. In Mycoplasma pneumonia infection group, IL-17 raises neutrophils to resist infection, NE%in BALF can indirectly reflect the mycoplasma infection degree.

Clinical and TUBB4A mutation analysis of hypomyelination with atrophy of the basal ganglia and cerebellum in a family / 中华实用儿科临床杂志

Objective To analyze the clinical data and TUBB4A mutation of hypomyelination with atrophy of the basal ganglia and cerebellum (HABC)in a family,thus to provide accurate genetic counseling and prenatal diagno-sis for this family with HABC,and also to provide clinical experience for the diagnosis of HABC in China.Methods The clinical data of the proband and her family members were collected at the Department of Pediatrics,Peking Univer-sity First Hospital,December 201 4,including medical history,physical signs,and brain MRI,biochemical tests and metabolic disease screening.The associated gene of hereditary leukoencephalopathy was screened for the proband and her family members were screened by targeting -high -throughput sequencing technology,and then the genetic varia-tions were verified by Sanger sequencing.With those detection methods,the gene mutation was confirmed,and then ge-netic features were analyzed.Results Clinical features were as follows:nystagmus as the first symptom,and motor and mental retardation,dystonia and ataxia followed.Brain MRI indicated hypomyelination of white matter and atrophy of the basal ganglia and cerebellum.The clinical diagnosis of HABC was established based on the clinical features and brain MRI features above.Genetics features showed that one novel TUBB4A c.974G >T heterozygous missense muta-tion was found from the proband,which caused an amino acid change from the Trp into Leu (p.Trp325Leu).Both of her parents with normal phenotype were of wild -type in this site.Conclusions The proband from this family was diagnosed clinically based on her clinical data.One novel TUBB4Ac.974G > T (p.Trp325Leu)was founded in this study.Therefore,the spectrum of TUBB4A mutation will be expanded.In addition,this study elucidated clinical and genetic characteristics in this family with HABC,which may lay a solid foundation for the accurate genetic counseling and prenatal diagnosis.This study reported the first case of HABC caused by TUBB4A mutation in China.

Effects of HIF-1αon epithelial-mesenchymal transition, invasion and migration in colorectal cancer cells / 天津医药

Objective To explore whether hypoxia could promote epithelial-mesenchymal transition (EMT) in various differentiated colorectal cancer cells, and analyse the effect of hypoxia on invasion and migration of colorectal cancer cells. Methods HCT116 (poorly differentiated) and HT-29 (highly differentiated) colorectal adenocarcinoma cells were selected respectively. The morphological changes of two cell lines were observed after 0,10,25,50,100 and 150 mg/L cobalt chloride (CoCl2) treatment for 48 h. The expression of hypoxia-inducible factor-1α(HIF-1α) protein was analysed after 0, 10,25,50,100 and 150 mg/L CoCl2 treatment for 48 h. An optimal concentration of CoCl2 was then selected. Methylthiazolyl tetrazolium (MTT) assay was used to detect the proliferation of two kinds of colorectal cancer cells induced by CoCl 2 at different time points (0, 24, 48, 72 and 96 h), and to select an optimal time. Under the optimal concentration and time conditions, the HCT116 and HT-29 cells were processed by hypoxia (hypoxia group) and normoxia (normoxic group). Transwell invasion assay and Wound healing assay were used to detect cell invasion and migration in two groups. Western blot assay and RT-PCR were used to detect protein and mRNA expression levels of HIF-1α, E-cadherin and Vimentin in two groups. Results Two kinds of cells showed obvious morphological changes after 50 mg/L CoCl2 treatment for 48 h. HIF-1αprotein level first increased and then decreased in two groups of cells with the increased concentration of CoCl 2, and 50 mg/L CoCl2 was the optimal concentration (P<0.05). The cell proliferation showed a tendency to decrease after the increase in both kinds of cells with or without hypoxia for 0-96 h (P<0.05), and 48 h was the optimal time. The transmembrane number and cell migration rate were significantly more in hypoxia group than those of normoxic group (P<0.05). The protein and mRNA levels of HIF-1α and Vimentin were significantly higher in hypoxia group than those of normoxic group in HCT116 and HT-29 cell lines (P<0.05). E-cadherin protein and mRNA levels were significantly lower in hypoxia group than those of normoxic group (P<0.05). Conclusion Hypoxia can promote EMT in different differentiated colorectal cancer cells, and can enhance invasion and migration of two kinds of colorectal cancer cells.

Analysis of pathogen in bronchoalveolar lavage lfuid and its clinical features in 80 children with lobar pneumonia / 临床儿科杂志

Objective To investigate the pathogenic types and clinical features of children with lobar pneumonia. Methods Eighty children with lobar pneumonia diagnosed from April 2013 to May 2015 were enrolled. Bronchoalveolar lavage lfuid (BALF) of patients were collected. FQ-PCR was used to detect and analyze pathogens in BALF. Results In 80 cases, 59 cases were Mycoplasma pneumoniae, and 2 cases were Chlamydia pneumoniae, 12 cases were Streptococcus pneumoniae, 1 case was Klebsiella pneumoniae, 8 cases were adenovirus, 1 case was respiratory syncytial virus, 14 cases were of mixed infection. The prevalence of MP lobar pneumonia in children of 7-14 years old age group were higher than that of other age groups. Conclusions The pathogen of children with lobar pneumonia varied from Mycoplasma pneumoniae, Chlamydia pneumoniae, bacteria, virus and so on, and Mycoplasma pneumoniae was the common pathogen in lobar pneumonia.

Influence of stem-like cells on EMT occurrence in mice with triple-negative breast can-cer and on their biological behavior / 中国肿瘤临床

Objective:To discuss the influence of ALDH1+and CD133+phenotypic breast cancer stem-like cells in TA2 triple negative breast cancer on promoting epithelial-mesenchymal transition (EMT) occurrence in TA2 mice with triple-negative breast cancer and on their biological behavior. Methods:Flow cytometry was performed to analyze the markers ALDH1 and CD133 in TA2 mice triple nega-tive breast cancer and breast cancer stem-like cells with ALDH1+, ALDH1?, CD133+, and CD133?phenotypes, which were sorted out. Then, the TA2 mice were inoculated with sorted tumor cells according to cell type. The mice were divided into ALDH1+, ALDH1?, CD133+, and CD133-groups. The tumor-growing conditions were observed. A tumor tissue was sliced for the immunohistochemical testing of ALDH1?, CD133?, and EMT-related Twist1, E-cadherin, and VE-cadherin proteins. The expression difference of breast cancer stem cell surface markers ALDH1 and CD133 in triple-negative breast cancer and EMT-related proteins Twist1, E-cadherin, and VE-cad-herin was analyzed. Results:The expression rates of breast cancer stem cell markers ALDH1 and CD133 in TA2 mice triple negative breast cancer were 31.2%and 6.5%, respectively. The tumor growth ability of TA2 mice from ALDH1+group was obviously stronger than that from ALDH1?group. The CD133+group was evidently stronger than CD133?group. The immunohistochemical results showed that ALDH1, Twist1, and VE-cadherin expression levels in the ALDH1+group were evidently higher than that in the ALDH1?group (all P<0.05). E-cadherin expression decreased (P<0.05). CD133?, Twist1, and VE-cadherin expression levels in CD133+group were higher than that in CD133?group (all P<0.05). Conclusion:In TA2 mice triple negative breast cancer, ALDH1+and CD133+phenotypic breast cancer stem-like cells may influence the expression of EMT-related proteins, and promote the formation of triple-negative breast cancer.

Clinical analysis of transcatheter closure treatment for the inaf nt wti h patent ductus arteriosus compli-cated with middle and severe pulmonary arterial hypertension / 中国小儿急救医学

Objective To evaluate the efficacy and safety of interventional occlsu ion operait no by analyizn g thes urgical data of 23 csa es of infants with patent ductusa rteriosus complicated with middle and severe pulmo an ry arterial hypertension.Methods Teh 23 cases of infants wiht patne t ductus arteriosus com-plicated with middlea nd severe pulmonary arterial hypertensionw ere collected in the hospital from January 2011 to December2014 .These infatn s rce eived transcateh ter occlusion with intravenuo s anesthesia after the preoperative examination.The operation procse s included:right ventriculography and pulmonary atr ery pressure tested,then lateral angiogar phy of descending aorta was performed to observe the type and size of patent ud ctus atr eriosus and measure ascending aorta,descending aortic pressure,and recorded the pressure re-spectively.1 ml blood sample of ascending aorta,pulmonary artery and inef rior vena vein respectively was used for gas analysis.All these data was used to calculate pulmonary vascular resistance.After tried to plug-ging effectiveyl we can release the occluder.In the postoperative 24 h,1 month,3 months,the infants should be measured with Doppler echocardiography,chest X ray and electrocardiogram examination.Results The clinical symptoms disappeared and the short-term follow-up was not associated with the complications of interventional therapy.Th e comparison of the pressure changes before and after the operation were performed as following, aortic per ssure decreased [ preoperation ( 68.3 ±17.5 )/( 21.4 ±3.7 ) mmHg, postoperation (52.4 ±8.7)/(15.6 ±3.5) mmHg,1 mmHg=0.133 kPa],ascending aorta pressure increased(preoperation (83.5 ±5.9)/(51.3 ±3.6) mmHg,postoperation(88.2 ±5.1)/(52.4 ±2.7) mmHg),and descending aorta pressure increased ( preoperation ( 81.4 ±3.3 )/( 48.2 ±2.7 ) mmHg, postoperation ( 86.5 ±4.7 )/(51.5 ±3.2) mmHg), the differences were statistically significant before and after surgery ( t =5.455/3.945 ,P<0.01;t=-2.696/-1.193 , P<0.05; t=-4.167/-3.745 , P<0.01 ) .Conclusion Under conditions of mastering the appropriate operation time and strengthening the management of the perioperative management,transcatheter measurement is safe and effective for infants with patent ductus arteriosus compli-cated with middle and severe pulmonary arterial hypertension.

Expression and prognostic significance of sodium-coupled neutral amino acid transporter 1 in esophageal squamous cell carcinoma / 第二军医大学学报

Objective To investigate the expression of sodium-coupled neutral amino acid transporter 1 (SNAT1) in esophageal squamous cell carcinoma (ESCC) tissues, and to evaluate its prognostic value in patients with ESCC. Methods The tissue microarray including 183 ESCC tissues and their corresponding adjacent tissues was subjected to immunohistochemistry detection and Western blotting analysis of SNAT1 expression, and their correlation with the clinicopatho logical parameters and overall survival time was analyzed. Results SNAT1 was highly expressed in ESCC tissues, with the positive rate being 45. 4% (83/183); we also noticed that t was not expressed in normal squamous epithelium tissues. Our analysis results indicated that overexpression of SNAT1 was significantly correlated with tumor size (P=0.023), lymph node metastasis (P=0.007) and tumor stage (P=0.003). The Kaplan-Meier analysis showed that patients with overexpression of SNAT1 had significantly worse outcome than those with negative SNAT1 expression (P=0.001). Cox regression analysis revealed that over expression of SNAT1 was an independent factor for prognostic of patients with ESCC (P=0.004). Conclusion The expression of SNAT1 is abnormally increased in ESCC tissue, indicating that SNAT1 expression may serve as an important biomarker for prognosis evaluation in patients with ESCC.

20 cases on arteriovenous fistula high clinical applications of / 中国综合临床

Objective To investigate an effective method of autologous arteriovenous fistula formation for the elderly uremic patients with diabetic nephropathy.Methods The nest in the elbow above the median cubital vein at the curved incision 1-2 cm,inline high Fistula,namely arm cephalic vein and brachial artery bifurcation of the upper portion of the distal radioulnar side anastomosis in 20 cases over 75 years elderly uremic patients with diabetic nephropathy.And compared the effect of 16 cases of patients with similar conventional fistula.Results Twenty cases over 75 years elderly uremic patients with diabetic nephropathy were successful in all high dynamic fistula intravenous,blood flow was (1 007±131) ml/m,the immediate opening rate of 16 patients similar to conventional instant fistula was 12.5％,and a year after the opening rate was 6.25％ Conclusion High fistula are more easy to shape conventional,easy mature,full blood flow characteristics comparing with commom fistula,so elderly uremic patients with diabetic nephropathy fistula making difficult situation has been effectively improved.

Relationship of different histological classifications of gastric cancer with microvessel density and related factors / 中国肿瘤临床

Objective:To investigate the correlations of Lauren classification and world health organization (WHO) classification of gastric cancer (GC) with microvascular density (MVD), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (VEGFR1), vascular endothelial growth factor receptor 2 (VEGFR2), and p53. Methods:The clinical data of 89 patients with GC were collected. The collected specimens were categorized on the basis of Lauren classification and WHO classification. CD34/periodic acid-Schiff (PAS) double staining was performed to validate MVD. Immunohistochemistry was conducted to investigate the expression levels of MMP-2, MMP-9, VEGF, VEGFR1, VEGFR2, and p53. Results:MVD was not correlated with Lauren classification or WHO classification (P>0.05). Lauren typing was associated with the expression levels of MMP-9, VEGFR1, and p53 (P0.05). Cox proportional hazards model revealed that Lauren classification and WHO classification were the prognostic factors of overall survival (P<0.05). Conclusion:This research on tumor related factors, angiogenesis, and different classifications of GC may provide new methods to treat this disease.

BMP4 promotes migration and invasion of hepatocellular carcinoma by inducing epithelial-mesenchymal transition / 中国肿瘤临床

Objective:To determine the expression of BMP4 in hepatocellular carcinoma (HCC) and to study the role of BMP4 in inducing epithelial-mesenchymal transition (EMT) to analyze the effect of BMP4 on the migration and invasion of HCC cells. Methods: The expression of BMP4 in HCC specimens was examined by immunohistochemistry staining, and the correlations were analyzed between the expression of BMP4 and clinicopathological data. The BMP4 expression plasmid was transfected into HepG2 cells to induce exogenous overexpression of BMP4 protein. The changes of HepG2 cell morphology were detected after BMP4 transfection by using a microscope; the changes of the expression of BMP4, EMT-related protein (E-cadherin, Vimentin) in HepG2 cells were detected by Western blot after transfection of BMP4;the wound healing assay in vitro was used to detect the effects of BMP4 gene transfection on the ability of migration of HepG2 cells;the invasion assay was used to determine the role of transfection of BMP4 on the invasive potential of HepG2 cells. Results: Immunohistochemistry staining method displayed that BMP4 expression was positively associated with age, histological differentiation, stage, and poor prognosis. After BMP4 overexpression, the morphology of HepG2 cells showed significant changes from a paving stone structure with cell-cell adhesion to a fibroblastic shape, which showed typical EMT change; Western blot exhibited that the expression of E-cadherin was downregulated and the Vimentin expression was upregulated in HepG2 cells;the wound healing and invasion assay showed that the migration and invasion potentials of HepG2 cells were significantly enhanced. Conclusion: BMP4, which displayed a high expression in HCC specimens, was closely associated with clinicopathologic data, and BMP4 may promote migration and invasion of HCC cells by inducing epithelial-mesenchymal transition.