Gene Analysis for the Sebaceous Carcinoma of Scalp by Whole Exome Sequencing / 中山大学学报(医学科学版)

ObjectiveTo reveal the differences of the related pathogenicity gene mutations between sebaceous adenocarcinoma （SC） of scalp and sebaceous adenoma （SA） of scalp on whole exome level. MethodsWhole exome sequencing was performed on a SC sample and a SA sample by Illumina Hiseq 2500 platform. Suspicious single nucleotide variation sites were selected for mutation conservation and functional analysis. SciClone was used to track subclone evolution and clonal map information was obtained for each tumor sample. The high-frequency significant gene mutations in the tumor sample were screened by MutSigCV software， and compared with the known driver genes. ResultsTwo driver genes TFDP1 and ACVR1B harboring mutations in scalp SC compared to SA were found. ConclusionsThe finding of mutation in driver genes TFDP1 and ACVR1B should be confirmed in a large cohort， which might reveal the mechanism of scalp SC development and find a therapeutic target for SC.

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

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

Correlation between Serum Osteoprotegerin/Soluble Receptor Activator of Nuclear Factor-κβ Ligand Levels and the Left Ventricular Diastolic Dysfunction in Type 2 Diabetes / 中山大学学报(医学科学版)

ObjectiveTo study the possible correlation between serum osteoprotegerin （OPG）/soluble receptor activator of the nuclear factor κB ligand （sRANKL） levels and the left ventricular diastolic dysfunction （LADD） in patients with type 2 diabetes mellitus （T2DM）. MethodsTotally 68 T2DM patients and 37 healthy controls were selected. Serum OPG and sRANKL were determined by solid-phase enzyme-linked immunosorbent assay （ELISA）. The left ventricular diastolic function of T2DM patients was measured by transthoracic echocardiography， where E/A < 1 were regarded as LVDD. T2DM patients were further divided into two subgroups according to E/A ratio （E/A≥1.0 and E/A<1）. Spearman correlation analysis， logistic regression and ROC curves were used to assess the possible correlation between serum OPG/sRANKL and LADD in T2DM patients. ResultsCompared with the healthy controls， serum OPG level in T2DM patients was higher with statistically significant difference （P <0.01）， while serum sRANKL level was lower without statistically significant difference （P =0.32）. T2DM patients with E/A<1 had significantly higher OPG level and lower sRANKL level than those with E/A≥1（P <0.01） in subgroup analysis. Spearman correlation analysis showed serum OPG level was negatively correlated with E/A ratio， while sRANKL was positively related with E/A ratio. In single factor logistic regression analyses， serum OPG ［OR （95% CI）=1.068 （1.031， 1.106）， P<0.001］ and sRANKL ［OR （95% CI）=0.976 （0.959， 0.992）， P=0.003］ were significant correlation with LVDD in T2DM patients. ROC curve analysis showed that the sensitivity and specificity of combined OPG and sRANKL in diagnosing T2DM patients LADD were 78.13% and 88.3%， respectively （area under the curve： 0.857； 95% CI=（0.768， 0.946）； P<0.001）. ConclusionsThe elevated OPG and decreased sRANKL levels may be associated with LADD in T2DM patients.

Free blood use among voluntary blood donors in Shiyan city / 中国输血杂志

【Objective】 To analyze the situation of free blood use and direct reimbursement of blood expenses in Shiyan city, so as to provide basis for more effective service of free blood use and direct reimbursement of blood expense after blood donation, and recruitment and retain of blood donors. 【Methods】 The data of free blood use from 1999 to 2019 in Shiyan city were collected for statistical analysis. Measurement data were described by mean±SD, and enumeration data were described by rate or construction ratio. Chi-square test was used for comparison of construction ratio or rate of stratified groups. 【Results】 In the past 21 years, free blood use has developed from the overall increasing stage (1999~2014) to the overall stable stage (2014~2019). Free blood use accounted for 3.52%(40 722.11/1 156 307.85)of the blood donation. Among the blood recipients, blood donors themselves accounted for only 18.83%(2 240/11 898), far lower than the proportion of their direct relatives. Since 2013, Shiyan has carried out the direct reimbursement of clinical blood expenses of voluntary blood donors according to the unified requirements of Hubei Province. The direct reimbursement rate over the past 7 years was 76.49%(5 397/7 056), and the proportion of people from other cities who applied for reimbursement was 4.02%(284/7 056). 【Conclusion】 The policy of free blood use and blood expense direct reimbursement has promoted the development of blood donation. It can also be further improved by relaxing the reimbursement scope of free blood use, strengthening publicity, optimizing services, strengthening supervision, using WeChat public service platform to carry out remote free blood, and strengthening national network construction.

Efficacy and Safety of Sacubitril-valsartan in the Treatment of Heart Failure without Reduced Ejection Fraction ： A Meta-analysis / 中国药房

OBJECTIVE：To systematically evaluate the efficacy and saf ety of sacubitril-valsartan in the treatment of heart failure without reduced ejection fraction （non-HFrEF）patients，and to provide evidence-based reference for its clinical treatment. METHODS：Retrieved from Cochrane Library ，PubMed，Embase，CNKI，VIP and Wanfang data ，during the inception to Feb. 29th，2020，randomized controlled trials （RCTs）about sacubitril-valsartan （trial group ）versus routine medicine as renin- angio- tensin converting enzyme inhibitors/angiotensin Ⅱ receptor antagonists （control group ） in the treatment of non-HFrEF were collected. After literature screening and data extraction ，the quality of included literatures were evaluated with Cochrane bias risk evaluation tool 5.3.0. Meta-analysis was conducted with Stata 14.0 software，and the publication bias analysis and sensitivity analysis were performed. RESULTS ：Totally 6 RCTs were included ，involving 5 502 patients. Results of Meta-analysis showed that the HF re-hospitalization rate [RR ＝0.84，95％CI（0.77，0.91），P＜0.001] and the serum creatinine elevation rate [RR ＝0.78，95％ CI（0.67，0.91），P＝0.001] in trial group were significantly lower than control group. NYHA classification improvement rate [RR ＝ 1.25，95％CI（1.10，1.43），P＝0.001] and the hypotension rate [RR ＝1.43，95％CI（1.24，1.65），P＜0.001] were significantly higher than control group. There was no statistical significance in the cardiovascular mortality [RR ＝0.94，95％CI（0.79，1.12）， P＝0.481]，all-cause mortality [RR ＝0.95，95％CI（0.83，1.08），P＝0.417]，the levels of NT-proBNP [WMD ＝－301.16，95％CI （－602.77，0.44），P＝0.050] and LVEF [WMD ＝1.49，95％CI（－1.33，4.32），P＝0.300] after treatment ，and the hyperkalaemia rate [RR ＝0.88，95％CI（0.77，1.01），P＝0.070] between 2 groups. The results of publication bias analysis and sensitivity analysis showed there was a high possibility of publication bias ，and the results of several indexes were not stable. CONCLUSIONS ： Sacubitril-valsartan may effectively reduce HF re-hospitalization rate and the risk of elevated serum creatinine in non-HFrEFpatients，improve the heart function but the risk of hypotension is high. The results should be interpreted carefully.

New progress in the treatment for gastrointestinal stromal tumors / 医学研究生学报

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors. Surgery remains to be the primary treatment for patients with localized GISTs, but there are still many patients suffering from tumor metastasis and recurrence after surgery. Imatinib adjuvant therapy plays an important role in the prevention and treatment of tumor metastasis and recurrence, but there are still many controversies in terms of dosage and time of administration. For initial unresectable GISTs with large volume, neoadjuvant therapy may considered to be an option. Sunitinib and regorafenib have played an important role in the second and the third line treatments. BLU-285 has brought hope to patients with mutation of PDGFRA D842. The emerging immunotherapy is still in the exploration stage of gastrointestinal stromal tumors in recent years. This article reviews the research progress of surgical treatment, neoadjuvant therapy, postoperative adjuvant therapy and other treatments for GISTs.

Study on the Optimization of the Extraction Technology of Shengmaiyin Polysaccharide and Its Regulation Effects on Intest- inal Function of Spleen Deficiency Model Rats / 中国药房

OBJECTIVE： To optimize the extraction technology of Shengmaiyin polysaccharide， and to investigate the regulation effects of Shengmaiyin and its polysaccharide on intestinal function of spleen deficiency model rats. METHODS： The contents of polysaccharide were determined by phenol-sulfuric acid method， and the extraction rate of polysaccharide was calculated. Using extraction rate of Shengmaiyin polysaccharide as investigation index， singel factor and orthogonal tests were used to optimize material-liquid ratio， extraction time， extraction temperature and extraction times of Shengmaiyin polysaccharide. Validation test was also conducted. Totally 80 male SD rats were randomly divided into blank group， model group， Shengmaiyin low-dose， medium-dose and high-dose groups （350， 700， 1 400 g/L， by crude drug）， Shengmaiyin polysaccharide low-dose， medium-dose and high-dose groups （24.5， 49， 98 g/L， by crude drug）， with 10 rats in each group. Except for blank group， other groups were given Rheum palmatum water decoction 10 mL/kg to induce spleen deficiency model， once a day， for consecutive 15 d. Since the 16th day， blank group and model group were given isovolumic water intragastrically， while other groups were given corresponding drugs， once a day， for consecutive 10 d. The general status of rats and body weights were recorded in each group. The serum contents of D-xylose， gastrin （GAS） and vasoactive intestinal peptide （VIP） were detected by phloroglucinol method or ELISA. RESULTS： The optimal extraction technology of Shengmaiyin polysaccharide was material-liquid ratio 1 ∶ 10（g/mL）， extraction time 45 min， extraction temperature 80 ℃， extracting for 1 time. Results of validation test showed that extraction rates of the polysaccharide in 3 times were 7.43％， 7.64％， 7.80％ （RSD＝1.01％， n＝3）. After modeling， except for blank group， other groups suffered from loose stools， thin body and reduced food intake， and the body weight and serum level of D-xylose were decreased significantly compared with blank group （P＜0.01）. After last medication， above symptoms of administration groups were improved to different extents. Except for model group， body weight and serum contents of D-xylose in other groups were increased significantly than those before modeling or before medication （P＜0.05 or P＜0.01）. Compared with blank group， body weight and serum content of GAS were decreased significantly in model group， while serum content of VIP was increased significantly （P＜0.01）. Compared with model group， body weight of Shengmaiyin medium-dose group and Shengmaiyin polysaccharide low-dose and high-dose groups， serum contents of D-xylose and GAS in Shengmaiyin medium-dose and high-dose groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were increased significantly， while serum contents of VIP in Shengmaiyin groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： The optimized extraction technology of Shengmaiyin polysaccharide is stable and feasible. Shengmaiyin and its polysaccharide contribute to the recovery of intestinal function of spleen deficiency model rat， the effects of which may be associated with the secretion regulation of GAS and VIP.

Blocking programmed death-ligand 1 attenuates maturation inhibition of dendritic cells by co-cultured breast cancer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study if programmed death-ligand 1 (PL-L1) expression in breast cancer cell activates PD-L1/PD-1 pathway in dendritic cells to inhibit dendritic cell maturation.</p><p><b>METHODS</b>Human monocytes were induced to differentiate into immature dendritic cells using GM-CSF and IL-4, and further to mature dendritic cells using TNF-α. PD-L1-expressing breast cancer cell line MDA-MB-231 was co-cultured in contact with the dendritic cells to observe the effects of the breast cancer cells on the maturation of the dendritic cells. A PD-L1 blocking antibody was applied to the co-culture, and the changes in the inhibitory effect of the MDA-MB-231 cells on dendritic cell maturation was observed. TNF-α-induced dendritic cells were treated with a recombinant human PD-L1 protein to study the effect of PD-L1/PD-1 pathway activation on the maturation of dendritic cells. The expression of PD-L1 in MDA-MB-231 cells and the dendritic cell maturation marker HLA-DR and CD83 were analyzed using flow cytometry.</p><p><b>RESULTS</b>MDA-MB-231 cell line showed PD-L1 positivity on the cell membrane cells at a rate as high as (99.7∓0.15)%. In mature dendritic cells, the positivity rates for HLA-DR and CD83 were (88.8∓6.96)% and (18.36∓3.07)%, respectively, but in the co-culture system, the positivity rates of the dendritic cells were significantly decreased to (42.76∓10.52)% (P<0.01) and (9.93∓2.74)% (P<0.05), respectively, indicating that MDA-MB-231 cells inhibited the maturation of dendritic cells. Following treatment with a PD-L1 antibody isotype control, the percentages of HLA-DR- and CD83-positive cells in the co-culture were (45.17∓10.19)% and (10.15∓2.54)%, which were significantly increased to (63.46∓1.72)% and (16.46∓2.58)% after treatment with PD-L1 antibody, respectively (both P<0.05). Compared with the mature dendritic cell controls, the cells treated with the recombinant human PD-L1 protein exhibited significantly lowered percentages of HLA-DR-positive [from (84.23∓4.18)% to (2.56∓2.39)%, P<0.05] and CD83-positive cells [(87.26∓1.54)% to (60.67∓1.63)%, P<0.05].</p><p><b>CONCLUSION</b>The effect of PD-L1 antibody therapy on triple negative breast cancer can be partially mediated by blocking PD-L1 expression on breast cancer cell membrane, which attenuates the inhibition of dendritic cell maturation in the cancer microenvironment.</p>

Efficiency and safety of drug-eluting stents and bare metal stents in elderly coronary artery disease： Meta-analysis / 中国介入影像与治疗学

Objective To evaluate the efficiency and safety of drug-eluting stents (DES) and bare metal stents (BMS) in elderly with coronary artery disease (CAD).Methods PubMed,Embase,Cochrane Library,CNKI and Wanfang databases were searched before October 28,2017.The pertinent randomized controlled trials (RCTs) and cohort studies (CS) were included.The outcomes were all-cause mortality (ACM),myocardial infarction (MI),target vessel revascularization (TVR),cardiac death (CD),stent thrombosis (ST) and bleeding.Relative risk (RR) with 95％ confidence interval (95％CI) was analyzed.Results A total of 17 studies with 10 813 patients were included.Overall and in CS group,there were significant lower risks of ACM (RR＝0.71,0.68,both P＜0.001),MI (RR＝0.66,P＜0.001;RR＝0.60,P＝0.008),TVR (RR＝0.50,0.54,both P＜0.001),CD (RR＝0.73,0.72,P＝0.003,0.020) and ST (RR＝0.68,0.59,P＝ 0.02,0.01) in DES patients compared with BMS patients,but the risk of bleeding was not significantly different (RR ＝ 1.00,1.00,P ＝ 0.96,0.97).In RCT group,DES (compared with BMS) was only associated with significantly lower risks of MI (RR＝0.68,P＝0.003) and TVR (RR＝0.43,P＜0.001),no difference was detected in other indexes.Conclusion DES is more efficacious and safer than BMS for elderly patients with CAD.

Effects of dexmedetomidine on expression of c-fos in hippocampus and dentate gyrus in a rat model of endotoxic shock / 中华麻醉学杂志

Objective To evaluate the effects of dexmedetomidine on the expression of c-fos in hippocampus and dentate gyrus in a rat model of endotoxic shock.Methods Thirty-five pathogen-free healthy male Sprague-Dawley rats,aged 3-4 months,weighing 250-300 g,were divided into 5 groups (n =7 each) using a random number table:normal saline group (group NS),dexmedetomidine group (group D),endotoxic shock group (group ES),low-dose dexmedetomidine plus lipopolysaccharide (LPS) group (group LD) and high-dose dexmedetomidine plus LPS group (group HD).Dexmedetomidine 0.5 μg/kg was injected via the tail vein in D and LD groups,and dexmedetomidine 4.5 μg/kg was given in group HD.Normal saline 0.5 ml/kg was injected in NS and ES groups,5 min later normal saline 0.5 ml/kg was injected in NS and D groups and LPS 5 mg/kg was injected in the other groups,and the injection time was 10 min in all groups.Rats were sacrificed at 6 h after LPS injection,brains were removed,and the hippocampus and dentate gyrus were isolated for detection of the expression of c-fos by immunohistochemistry.Results Compared with group NS or group D,the expression of c-fos in the hippocampus and dentate gyrus was significantly up-regulated in group ES (P＜0.05).Compared with group LPS,the expression of c-fos in the hippocampus and dentate gyrus was significantly down-regulated in LD and HD groups (P＜0.05).Compared with group LD,the expression of c-fos in hippocampal CA1 and CA3 areas was significantly down-regulated in group HD (P＜0.05).Conclusion The neuroprotective mechanism of dexmedetomidine is related to inhibiting the up-regulated expression of c-fos in the hippocampus and dentate gyrus in a rat model of endotoxic shock.

Reference range of left ventricular strain measured by three-dimensional speckle-tracking imaging:Meta analysis / 中国介入影像与治疗学

Objective To obtain the normal reference ranges of global longitudinal strain (GLS),global circumferential strain (GCS),global area strain (GAS) and global radial strain (GRS) of left ventricular in normal adults by three-dimensional speckle tracking imaging (3D-STI) using Meta analysis.Methods Eligible trials which detected global strain of left ventricular in normal subject through 3D-STI were searched in Embase,Pubmed,Cochrane Library database.According to the heterogeneity,parameters of contained studies were analyzed the weighted mean difference (WMD) and 95％ confidence interval (CI).The statistical software was STATA 12.0.Results Totally 1 552 healthy adults from 27 articles were included.Based on the Meta-analysis,theWMDand 95％CIofGLSwere 17.80 and (16.27,19.33),of GCS were 24.73 and (22.50,26.95),of GRS were 47.86 and (39.52,56.19),of GAS were 36.17 and (34.08,38.26).Conclusion The Meta analysis defines reference range of strains obtained by 3D-STI in healthy adults.Using these parameters of 3D globe strains,a guidance of reference for patienfs management and therapy selection may be provided.

Molluscicidal effect of suspension concentrate of niclosamide ethanolamine salt / 中国血吸虫病防治杂志

Objective To evaluate the molluscicidal effect of suspension concentrate of niclosamide ethanolamine salt(SC-NE)against Oncomelania hupensis snails in laboratory and field. Methods The experiment of SCNE against the snails by using the immersing and spraying methods was performed in laboratory and field,with control groups of wettable powder of ni-closamide ethanolamine salt(WPN). Results In the laboratory,LC50(s) of SCNE for 24,48 h and 72 h by using the immersion method were 0.0926,0.0629 mg/L and 0.0549 mg/L,respectively. The mortality rates of snails for 24,48 h and 72 h by using the immersion method were all 100％ with the concentrations of 0.25 mg/L. The mortality rates of snails were all 100％ while spraying SCNE for 3 d in the laboratory with the concentrations of 0.25 g/m2. In Jiangling County,except 0.5 g/m3 SCNE immers-ing the snails for 24 h,the mortality rates of snails by using SCNE with the immersing method were all 100％. While the concen-tration of SCNE was 0.5 g/m3 or above,the mortality rates were all 100％after the use of it with the immersion method for 2 d in Gong'an County. In Jiangling County,the mortality rates of snails by using SCNE 0.5 g/m3 for 1 d,3 d,and 7 d with the spray-ing method were 87.5％,92.82％and 97.40％respectively. While the concentration of SCNE was 0.5 g/m3,the mortality rates were 85.94％,86.78％and 94.21％respectively after the use of it with the spraying method for 1 d,3 d,7 d in Gong'an Coun-ty,and the molluscicidal effect of SCNE(1.0 g/m2)was higher than that of WPN. Conclusion SCNE has a high molluscicidal effect in the laboratory and field,and it is a novel and simple formulation of niclosamide.

Comparative study of clinical efficacy between the Solitaire mechanical thrombectomy and selective arterial thrombolysis in treatment of acute cerebral infarction / 局解手术学杂志

Objective To investigate the clinical safety and efficacy of mechanical thrombectomy(MT) with Solitaire stent compared with the selective intra-arterial thrombolysis (IAT) in the treatment of acute cerebral infarction.Methods Totally 82 cases patients with severe acute ischemic stroke caused by middle cerebral artery stenosis from January 2014 to May 2016 in the stroke treatment center of the first people's Hospital in ZunYi city who were applied with the mechanical thrombectomy or the selective intra-arterial thrombolysis was included,and a comparative analysis was conducted on the mTICI rating to assess the interventional recanalization,the NIHSS score after the treatment,bleeding rate,and conditions of neurological functional recovery 90 days after operation.Results There were 42 cases applied with the mechanical thrombectomy (MT group),40 cases applied with the intra-arterial thrombolysis (IAT group).The total effective rate reached 85.70％ in the MT group and 62.50％ in the IAT group,the difference was significant (P ＜ 0.05).Compared with NIHSS score before operation,the score after the treatment showed a decreased trend.And the NIHSS score of MT group was better than that of the IAT group in a week after the treatment (P ＜ 0.05).The bleeding rate was lower in the MT group with statistically significant difference (P ＜ 0.05).Conclusion Compared to IAT,MT can provide broader time window,higher recanalization rate and better outcome in patients with severe acute ischemic stroke.

System practice, experience and trends on the long-term care insurance of China: Based on a comparative study of Qingdao model and Nantong model / 中国卫生政策研究

The system of long-term care insurance,which is one supportive system for the increasing long-term care needs of the elderly in China will be the highlight for the construction of social security system during the 13th Five-Year Plan period.This paper focuses on the current background,models feature and system design of regional practice of long-term care insurance in Qingdao and Nantong,and makes a comparison between them.On the basis of the summarized experience and inspiration,and the existing problems as well as the experience in advanced countries,the author puts forward the corresponding suggestions,in the hope of providing valuable references for the toplevel design of the long-term care insurance system.

System practice, experience and trends on the long-term care insurance of China: Based on a comparative study of Qingdao model and Nantong model / 中国卫生政策研究

The system of long-term care insurance,which is one supportive system for the increasing long-term care needs of the elderly in China will be the highlight for the construction of social security system during the 13th Five-Year Plan period.This paper focuses on the current background,models feature and system design of regional practice of long-term care insurance in Qingdao and Nantong,and makes a comparison between them.On the basis of the summarized experience and inspiration,and the existing problems as well as the experience in advanced countries,the author puts forward the corresponding suggestions,in the hope of providing valuable references for the toplevel design of the long-term care insurance system.

Genetic analysis of three families affected with split-hand/split-foot malformation / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the genetic etiology of three families affected with split-hand/split-foot malformation (SHFM).</p><p><b>METHODS</b>Peripheral venous blood samples from 21 members of pedigree 1, 2 members of pedigree 2, and 2 members of pedigree 3 were collected. PCR-Sanger sequencing, microarray chip, fluorescence in situ hybridization (FISH), real-time PCR, and next-generation sequencing were employed to screen the mutations in the 3 families. The effect of the identified mutations on the finger (toe) abnormality were also explored.</p><p><b>RESULTS</b>Microarray and real-time PCR analysis has identified a duplication in all patients from pedigrees 1 and 3, which have spanned FKSG40, TLX1, LBX1, BTRC, POLL and FBXW4 (exons 6-9) and LBX1, BTRC, POLL and FBXW4 (exons 6-9) genes, respectively. A missense mutation of the TP63 gene, namely c.692A>G (p.Tyr231Cys), was found in two patients from pedigree 2. FISH analysis of chromosome 10 showed that the rearrangement could fita tandem duplication model. However, next-generation sequencing did not identify the breakpoint.</p><p><b>CONCLUSION</b>The genetic etiology for three families affected with SHFM have been identified, which has provideda basis for genetic counseling and guidance for reproduction.</p>

A strategy for identifying six species of Sect. Cruciata (Gentiana) in Gansu using DNA barcode sequences / 药学学报

Located in the transition zone between the Qinghai-Tibet Plateau and the Loess Plateau, Gansu province is one of the distribution centers of Sect. Cruciata, Gentiana (Gentianaceae) in China. Six species in the section, G. crassicaulis, G. straminea, G. siphonantha, G. officinalis, G. dahurica and G. macrophylla, are native to Gansu. In this paper, samples of 6 species and Halenia elliptica (outgroup) were collected. Nuclear DNA ITS, chloroplast DNA matK, rbcL, rpoC1, trnL (UAA) intron, psbA-trnH, atpB-rbcL, trnS (GCU)-trnG (UCC), rpl20-rps12 and trnL (UAA)-trnF (GAA) were sequenced from these samples. Based on the sequence analyses, high intragenomic polymorphisms were detected in ITS regions of G. crassicaulis, G. straminea, G. siphonantha, G. officinalis and G. dahurica, and they showed incomplete concerted evolution. A methodological study to identifying such close-related species as G. macrophylla, G. officinalis and G. dahurica was carried out based on the special genotypes. The results showed that 7 cp DNA sequence fragments could be used to identify G. crassicaulis, G. straminea and G. siphonantha. With nr ITS genotype II, III and IV of G. dahurica, the species can be distinguished from the close-related G. officinalis using 12 cloned sequences in a sample (with statistical significance). The cp DNA sequences of G. macrophylla were classified into two genotypes, and with genotype II, the species can be distinguished from the close-related G. officinalis and G. dahurica using 6 test samples each (with statistical significance). Furthermore, DNA barcode sequences were determined for all 6 species in Gansu. Also, the studies provide some basic data for analyses of genetic diversity and identification of Gentiana species.

Phosphorylation of ezrin Tyr477 is critical in invasion of breast cancer cells stimulated by precursor of nerve growth factor / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of precursor of nerve growth factor (proNGF) in promoting invasion of breast cancer cells and its relation with ezrin expression and phosphorylation of ezrin Thr567 and Tyr477.</p><p><b>METHODS</b>Human breast cancer cell lines MDA-MB-231 and MCF-7 were stimulated by gradient concentrations of proNGF (0, 2.5, 5 and 10 ng/mL) for 16 h, and the invasion of the cells was assessed with Transwell assay. The expression of ezrin and the phosphorylation of ezrin Thr567 and ezrin Tyr477 in the treated cells were examined by Western blotting. MDA-MB-231 cells were transfected with pEnter-His-ezrinY477F (a dominant negative mutant) to study the role of phosphrylation of ezrin Tyr477 in the invasion of breast cancer cell stimulated by proNGF.</p><p><b>RESULTS</b>proNGF significantly promoted MDA-MB-231 and MCF-7 cell invasion in a concentration-dependent manner (P<0.05), and concentration- and time-dependently increased the phosphorylation of ezrin Tyr477 (P<0.05) without affecting the expression of ezrin or the phosphorylation of ezrin Thr567. The specific inhibitor of src, SKI-606, significantly inhibited the phosphorylation of ezrin Tyr477 induced by proNGF. Transfection with pEnter-His- ezrinY477F inhibited proNGF-induced invasion and phosphorylation of ezrin Tyr477 in MDA-MB-231 cells (P<0.05).</p><p><b>CONCLUSION</b>Phosphorylation of ezrin Tyr477 plays a critical role in the invasion of breast cancer cells stimulated by proNGF via proNGF/src/ezrin Tyr477 pathway.</p>

Nanog promotes the invasion of breast cancer cells by increasing PKCε expression / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the relationship between Nanog-promoted metastasis of breast cancer and ezrin(T567) phosphorylation, and explore the possible mechanism by which Nanog regulates ezrin(T567) phosphorylation.</p><p><b>METHODS</b>A siRNA construct targeting Nanog was transfected in breast cancer cells to knock down Nanog expression, and the changes in the cell invasion was detected using Transwell assay. The expression levels of Nanog and PKC and the phosphorylation level of ezrin(T567) were detected using Western blotting and immunofluorescent staining; the protein interaction between PKCε and ezrin was assayed by co-immunoprecipitation and Western blotting.</p><p><b>RESULTS</b>Nanog knockdown significantly decreased the expression of PKCε protein, phosphorylation level of ezrin(T567) and the invasion ability of breast cancer cells. PKCε knockdown obviously decreased the phosphorylation level of ezrin(T567) in the cells, and PKCε and ezrin were co-immunoprecipitated.</p><p><b>CONCLUDIONS</b>Nanogcan can upregulate the expression of PKCε to promote the phosphorylation of ezrin(T567), which can be a new mechanism by which Nanog promotes tumor metastasis.</p>

DNA-based identification of Gentiana robusta and related species / 中国中药杂志

The alpine plant Gentiana robusta is an endemic species to the Sino-Himalayan subregion. Also, it is one of the original plants used as traditional Tibetan medicine Jie-Ji. We sequence the nuclear ribosomal internal transcribed spacer (ITS) regions, matK, rbcL, rpoC1, trnL (UAA), psbA-trnH, atpB-rbcL, trnS( GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF( GAA) fragments of cp DNA in both G. robusta and such relative species as G. straminea, G. crassicaulis and G. waltonii. With Halenia elliptica as the outgroup, molecular systematic analysis reveals that G. robusta is a natural hybrid. G. straminea is the mother of hybrids, but the father is not very clear. In addition, the molecular markers for distinguishing G. robusta from the parental species or closely related species are identified, respectively. Our studies may provide valuable reference for the species identifications of medicinal plants with complex genetic backgrounds.