Objective To investigate the characteristics and possibility of using an image analyzer-aided method to count axotomized retinal ganglion cells (RGCs). Methods The left optic nerves of 18 rats were transected intraorbitally and a piece of gelform soaked in 5% fluorogold was applied to the ocular stump to retrogradely label the surviving RGCs. All animals were executed 2, 7 or 14 days after the operation (n=6 for each time point), respectively. The left retinae were removed, post-fixed and whole-mounted on the slides. The numbers of labeled RGCs were counted using both the conventional sampling method and image analysis, and compared statistically between the two methods. Results The number of surviving RGCs decreased sharply [(12 0663?9 089), (59 285?17 071) and (17 802?19 84) cells/mm 2 for image analyzer-aided method, and (118 237?7 898), (57 648?14 533) and (18 070?1 461) cells/mm 2 for conventional sampling method] when the survival time increased from 2 to 7 and 14 days. No significant difference was detected between the two groups at any corresponding time points. Conclusion The image analyzer-aided method is convenient, objective and reproducible, which can be used in the studies where counting RGCs is needed.

Objective To analyze the clinical and imaging characteristics of congenital sensorineural hearing loss (CSNHL)children combined with white matter (WM)lesions in order to provide evidence for clinical practice. Methods With referral to the Department of Pediatrics,Peking University First Hospital from November 201 1 to De-cember 201 5,documents of 78 patients of CSNHL combined with WMlesions were collected and analyzed for the clini-cal and imaging characteristics.Results Bilateral severe -profound hearing loss existed in all 78 cases,48.1 %(25 /52 cases)of the patients exhibited gross motor development delay,98.1 %(51 /52 cases)of them had normal cognition development.One hundred percent (61 /61 cases)of patients had abnormal language development.Infection occurring during pregnancy existed in 21 .2%(1 1 /52 cases)of the patients,the premature and smaller for the gestational age in-fants accounted for 28.9% (1 5 /52 cases).The bilateral multiple WMlesions from the brain MRI were in dot to flake sizes with sharp boundary,the intensity of T1 -weighted imaging decreased,T2 -weighted imaging and fluid attenuated inversion recovery increased.Eighty -two point one percent (64 /78 cases)of the patients were found to have the periventricular and subcortical WM involvement.The most frequently affected periventricular region was the posterior horn (91 .9%,68 /74 cases),followed by the anterior horn and temporal horn,and the least with the body involvement. The former three had a combined lesion tendency (55.4% -68.9%).There was an extensive involvement in the sub-cortical WMof parietal,frontal,temporal and occipital lobes respectively(73.5% -88.2%).Subcortical WM involve-ment of multiple lobes was common (accounted for 67.6% -77.9%).The enlargement of bilateral ventricles existed in 37.2%(29 /78 cases)of the patients and cystic changes in the subcortical WM of anterior temporal lobe could be found in 9.0% (7 /78 cases)patients.Calcification in 2 CT cases was reported.Corpus callosum and basal ganglia of all cases were normal.For cases with MRI scans more than once,WMlesions of 96.0%(24 /25 cases)patients became silent or self -restored.Conclusions The clinical presentations of CSNHL combined with WM lesions are mild,not paralleled with their multiple foci.It is considered as demyelination or a delay of myelination.Due to its benign course, it is probably not the contraindication for the cochlear implantation.

To investigate the effects of Astragalus membranaceus and Potentilla discolor mixture (APM) on insulin resistance (IR) and mRNA expressions of IR-related genes, including phosphatidylinositol 3-kinase (PI3-K), phosphoenolpyruvate carboxykinase (PEPCK) and peroxisome proliferator-activated receptor γ coactivator 1 (PGC1) in KKAy mice with early type 2 diabetes and to explore the gene regulation mechanisms of AMP.

To observe the effects of tetramethylpyrazine (TMP) on the proliferation and type I collagen synthesis of rat cardiac fibroblasts (CFBs) induced by angiotensin II (Ang II), and to explore the mechanism of TMP in treating myocardial fibrosis.

To explore the effects of Shengmai injection and Xuesaitong injection, compound Chinese herbal medicines for replenishing qi and activating blood, on ventricular fibrillation threshold, heart structure and connexin 43 (Cx43) expression in rats with myocardial infarction (MI).

Currently,a physiologically based pharmacokinetic(PBPK)model plays a key role in pharmaceutical research,which has been widely used at each stage of drug discovery and develop?ment. In the process of drug discovery,the selection of drug candidates is finished using the PBPK model to predict the pharmacokinetic properties of the drugs. In the process of preclinical development , through a combination of in vitro and physiological data amplification coefficient,the PBPK model can be used to predict not only the overall pharmacokinetic behavior of drug candidates in humans and animals and in vitro metabolism experiments,but also drug-drug interactions(DDI). In the course of clinical development,the PBPK model can help predict the difference between reference populations (age,different disease state,and polymorphism),especially the dosage and sampling time of the children. At present,the input parameters of PBPK model are mostly the mean values of the population,making it difficult to serve individuals. It is hoped that the input parameters of the model can reflect more of the individual characters according to the individual requirement,and that the time parameters of the input accord more with the actual physiological condition. In this article ,we briefly introduced the characteristics of common PBPK software,and reviewd the principle and feature of the PBPK model,as well as its application to drug discovery,preclinical development and clinical development,DDI,and individualized medication.

Protein phosphorylation is one of the most common post-translational modifications (PTMs)in various organisms,which plays critical roles in the regulation of intracellular biological processes,such as cell prolifera-tion,signal transduction,metabolismis and tumorigenesis.However,the low abundance of phosphoprotein in the biological systems poses significant challenges of current analytical techniques.In order to further understand the phosphoproteomics,the roles of phosphorylated proteins in life process,discovery of biomarkers,diagnosis and treatment of disease,enrichment strategies of high efficiency have been developed,including the design of new nanomaterials and combination of a variety of analytical methods,et al.In this paper,we reviewed the develop-ment of enrichment strategies for phosphoproteomics and application of phosphoproteomics in disease.

Objective To observe the effects of Fuzheng Huayu Capsule on the expressions of the main roles, Rock1 and Rock2, in RhoA/Rock signal transduction pathway in rats with myocardial infarction (MI);To explore the possible mechanism of Fuzheng Huayu Capsule to improve ventricular remodeling in myocardial fibrosis. Methods The MI model was induced by ligation of the left coronary artery. Rats with MI were randomly divided into the model group, Fuzheng Huayu group and Fasudil hydrochloride group. A sham-operation group threading without ligation was setting as a control group, with eight rats in each group. The rats were treated with corresponding medicine for 4 weeks from the second day after modeling. The expression of Rock1, Rock2 and its mRNA were detected by immunohistochemical and real-time PCR method. Results The protein expression of Rock1 and Rock2 in model group were significantly higher than those in the sham-operation group (P<0.05). The protein expression of Rock1 and Rock2 in the Fuzheng Huayu group and Fasudil hydrochloride group were lower than those in the model group. The mRNA expression of Rock2 was significantly higher in the model group than that in the sham-operation group (P<0.05). The mRNA expression of Rock1 in the Fasudil hydrochloride group was lower than that in the model group (P<0.05). The mRNA expression of Rock2 in the Fuzheng Huayu group and Fasudil hydrochloride group was lower than that in the model group (P<0.05). Conclusion Fuzheng Huayu Capsule can decrease the expression of Rock1, Rock2 and Rock2 in the marginal zone of myocardial infarction in rats with MI. The anti ventricular remodeling mechanism of Fuzheng Huayu Capsule maybe related with this.

OBJECTIVETo investigate the expression of placenta-derived RASSF1A gene in maternal plasma during first and second trimesters, and to explore its value for the prediction of pre-eclampsia.

METHODSFor 325 pregnant women of the first trimester, free DNA of plasma samples was extracted at 7-12, 13-18, and 19-24 gestational weeks, respectively. Methylation-sensitive restriction enzyme digestion followed by fluorescence quantitative PCR (MSRE+ PCR) was employed for analyzing the concentrations of hypermethylated RASSF1A gene. Blood pressure, proteinuria and clinical feature were monitored at the same time. Those who had subsequently developed pre-eclampsia were selected as the pre-eclamptic group, 30 normal pregnant women were selected as the control group. Hypermethylated RASSF1A gene in maternal plasma was retrospectively analyzed. The relationship between clinical classification, type of pre-eclampsia and concentrations of the gene were further analyzed.

RESULTSTwenty-six out of the 325 pregnant women developed pre-eclampsia as their only complication. At 13-18 gestational weeks, the mean concentrations of fetus-specific RASSF1A sequences were 141.62 copies/mL in maternal plasma of pre-eclamptic pregnancies, which was significantly greater than that of the controls (98.90 copies/mL). Fetus-derived RASSF1A levels were 2.03 fold higher in pre-eclamptic subjects than controls at 19-24 gestational weeks. There was a significant difference in the level of hypermethylated RASSF1A gene between the mild and severe pre-clamptic subjects at 13-24 gestational weeks (P< 0.05). The concentrations of the sequences were significantly higher in early-onset severe pre-eclampsia than late-onset severe pre-eclampsia at 19-24 gestational weeks (P< 0.05).

CONCLUSIONAltered expression of hypermethylated RASSF1A gene may be detected in maternal plasma during second trimester, which has important significance for early prediction of pre-eclampsia.

Female ; Gestational Age ; Humans ; Placenta ; metabolism ; Pre-Eclampsia ; blood ; diagnosis ; genetics ; metabolism ; Predictive Value of Tests ; Pregnancy ; Pregnancy Trimester, Second ; Prenatal Diagnosis ; methods ; Tumor Suppressor Proteins ; blood ; genetics

Triple negative breast cancer is a subtype of breast cancer with poor prognosis and lack of effective treatment. Cyclin dependent kinase (CDK) 4/6 inhibitors promote antitumor immunity by influencing the triple negative breast cancer immune microenvironment, such as increasing the tumor cell surface pragrammed death-ligand 1 protein expression, enhancing T cell activation and antigen presentation, changing the proportion of T cell subgroup and inducing lymphocyte infiltration. The change of immune microenvironment is related to tumor progression, but its mechanism is extremely complex. Exploring the mechanism of CDK4/6 inhibitor affecting immune microenvironment and its biomarkers can provide a new direction for the diagnosis and treatment of triple negative breast cancer.