In order to adapt to the requirements of modem medical knowledge and skills for higher medical workers,and to cultivate medical students' molecular medicine quality and comprehensive ability,Weifang Medical University broke the boundaries of disciplines and constructed experimental course group in molecular medicine based on the ideas of curriculum group construction.Molecular medical knowledge was integrated into the teaching process of experimental course group,experimental teaching content system was reasonably integrated and optimized,high quality teaching team was set up,multi-level experimental teaching platform was built and student-centered teaching mode was implemented to explore the experimental teaching approach which helped medical students to form systematic molecular medicine knowledge structure and ability structure.

ObjectiveTo observe the effects of salmeterol fluticasone on the glucose metabolism and bone density of the elderly patients with chronic obstructive pulmonary disease (COPD) combined with type 2 diabetes mellitus (T2DM).MethodsThirty-one patients with COPD combined with T2DM were divided into 2 groups by random digits table,14 cases in control group were given conventional therapy,17 cases in experimental group were given conventional therapy and fluticasone propionate 50/500 μg,twice a day,for 3 months.The fasting blood glucose,postprandial 2 hours glucose,glycosylated hemoglobin,plasma cortisol and bone density respectively before and after treatment were detected.ResultsIn experimental group,the fasting blood glucose,postprandial 2 hours glucose,glycosylated hemoglobin and bone density before treatment was (5.25 ± 0.21 ) mmol/L,(7.14 ± 0.33 ) mmol/L,(5.58 ± 0.26 )％,( 1.96 ± 0.11 ) g/cm2,and after treatment was(5.31 ± 0.27 ) mmol/L,(7.22 ± 0.29 ) mmol/L,(5.67 ± 0.23 )％,(2.03 ± 0.15 ) g/cm2,there was no significant difference (P ＞ 0.05).In control groups,the fasting blood glucose,postprandial 2 hours glucose,glycosylated hemoglobin and bone density before treatment was (5.33 ± 0.35) mmol/L,( 7.26 ± 0.29 ) mmol/L,( 5.62 ± 0.19 )％,( 1.88 ± 0.20 ) g/cm2,and after treatment was ( 5.36 ± 0.31 ) mmol/L,(7.30 ± 0.35 ) mmol/L,( 5.69 ± 0.26 )％,( 1.98 ± 0.17 ) g/cm2,there was no significant difference (P ＞ 0.05 ).There was no significant difference in plasma cortisol before and after treatment in two groups (P ＞0.05).ConclusionInhaling salmeterol fluticasone for elderly patients with COPD combined with T2DM is safe.

Objective Serologic proteome analysis method (SERPA) was used to compare the different of the serum proteins between normal and pancreatic cancer patients' serum, and to find a new specific pancreatic cancer marker. Methods HPLC was used to eliminate albumin from the serum, two-dimensional electrophoresis was used to separate the proteins. After imaging collection and analysis, the different proteins between pancreatic cancer and normal subjects were cut for mass spectrometry. Results Four discrepancy proteins were obtained. Guanylate cyclase-activating protein 2 was highly expressed in normal serum but not pancreatic cancer. Hp2-alpha, transthyretin and KIAA1018 protein were highly expressed in cancer patients'serum but not normal people. Conclusions KIAA1018 may become a promising tumor marker for screening and early diagnosis of pancreatic cancer.

Objective To analyze the mutations of causal genes in sixteen Chinese patients with suspicious Bartter syndrome,and follow up their treatment results.Methods Mutations were identified by the next generation sequencing and the multiplex ligation-dependent probe amplification (MLPA).Clinical and biochemical features at the first presentation as well as follow-up results were reviewed.Results 15 different CLCNKB gene mutations were identified in sixteen patients with BS,including 11 novel ones.A novel missense mutation and a novel small deletion were found from SLC12A1 gene.A novel gross deletion was found in CLCNKA gene.A recurrent missense mutation was identified from BSND gene.The whole gene deletion mutation of CLCNKB gene was the most frequent mutation (32％),and the rate of gross deletion was up to 50 percent in this group of Chinese patients.The most common clinical manifestations were development retardation (15/16),polydipsia and polyuria (15/16).All of the patients were detected with hypokalemia,hypochloremia and metabolic alkalosis.Indomethacin treatment had significant improvement to the stature and weight restoration.Conclusion The present study has found 19 mutations,including 14 novel ones,which enriches the human gene mutation database (HGMD) and provides valuable references to the genetic counseling and diagnosis of Chinese population.

Objective To investigate the relationship between CYP3A5 gene polymorphism and tacrolimus concentration/dose ratio in children living donor liver transplantation and the correlation with clinical efficacy,for the relatives living donor liver transplantation in children tacrolimus individualized medication providing reference indicators.Methods Peripheral blood samples were collected from children with relatives living donor liver transplantation in the center of liver transplantation,the genotype of CYP3A5 was determined by polymerase chain reaction (PCR)/pyrophosphate sequencing,The dosage of tacrolimus and blood concentration,liver and kidney function and other related indicators were measured within 3 months after operation According to genotypes,the children can be divided into gene expression group (CYP3A5 *1/*1 and CYP3A5 *1/*3) and non-expression group (CYP3A5*3/*3).The drug concentration (C0),tacrolimus dose / body weight (D/W) ratio,drug concentration/dose (C0/D) ratio of each genotype at 1 day,3 d,5 d,7 d,14 d,28 d,2 months and 3 months after administration and the genotype at the time point on liver and kidney function was carried out statistics.Results Among the 80 cases,36 cases (45.0％) were CYP3A5*3/*3,37 cases (46.2％) were CYP3A5*1/*3,7 cases (8.8％) were CYP3A5*1/*1.CYP3A5 gene expression group reached a therapeutic concentration range (C0 ＞ 8 μg/L) than the gene non-expression group takes longer time.There was no significant difference in CYP3A5 gene expression group between the non-expression group on the initial dose (P＞ 0.05);CYP3A5 gene expression group than the non-expression group,tacrolimus C0 within 1 month after operation were statistically significant.CYP3A5 gene expression group than the non-expression group,tacrolimus D/ W in addition to the first day after surgery,other time points were statistically significant (P＜0.05).CYP3A5 gene expression group than the non-expression group,C0/D at the above time points were statistically significant (P＜0.05);There were no significant differences in liver and kidney function between the two genotypes (P ＞ 0.05),but there was significant difference in the alkaline phosphatase.Conclusion CYP3A5 gene expression in children than non-expression group of children need higher doses to reach the therapeutic drug concentration;CYP3A5 gene polymorphism had significant effects on early tacrolimus C0,D/W and C0/D values;CYP3A5 gene polymorphism is instructive for the administration of tacrolimus in children with living donor liver transplantation,CYP3A5 gene type tests should be regular,improve efficacy and reduce the incidence of adverse reactions.

Objective To investigate the genetic characteristic of whole-genome of influenza A/H3N2 viruses in severe acute respiratory infection cases in Beijing area.Methods From 2014 to 2016,the viral RNA was extracted from 32 strains isolated from SARI cases,then sequenced by Ion Torrent PGM Sequencer.The phylogeny and molecular features of whole-genome were analyzed by Mega and Consurf software.Results The HA gene of tested strains isolated in 2014-2015 influenza season belonged to lineage 3C.3a and 3C.2a,while those isolated in 2015-2016 influenza season belonged to cluster 3C.2a.Moreover,compared with the vaccine strains,7 variant amino acids of protein of HA1 were identified,and two of them were located in antigenic sites.All isolates were sensitive to neuraminidase inhibitors while showed resistance to blockers for M2 ion channel.Conclusion The phylogenetic features of isolates studied in this study are similar with that of current circulating strains.However,the difference between isolates and vaccines should not be overlooked.

Recent studies have revealed that human milk harbors stem cells at various stages of differentiation,including pluripotent stem cells,hematopoietic stem cells and mesenchymal stem cells.The stem cellular components of human milk with high interindividual variability are affected by gestational age and lactation stages.The maternal stem cells which pass to the infant through human milk could help boost offspring growth and development.Human milk stem cell with multi-directional differentiation potential may be a new source of stem cell replacement therapy in the future.We hereby review the latest progress in human milk stem cells from these aspects.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cohort Studies ; Female ; Humans ; Male ; Middle Aged ; Pleural Effusion ; diagnosis ; pathology ; surgery ; Thoracoscopy ; methods ; standards ; Young Adult

BACKGROUND AND OBJECTIVETransbronchial needle aspiration (TBNA) and endobronchial ultrasound-guided TBNA (EBUS-TBNA) have been applied to the diagnosis for mediastinal lymph nodes. The aim of this study is to evaluate the clinical value and safety of TBNA and EBUS-TBNA on hilar and mediastinal lymph nodes of lung cancer patients.

METHODSTwo hundred fifty patients with suspected lung cancer were enrolled. All petients with hilar and/or m lymphoadenopa-ediastinal thy found by CT scan received TBNA, biopsy and brushing. EBUS-TBNA was performed in 15 patients among them.

RESULTSLung cancer were confirmed in 180 patients by TBNA, biopsy and brushing. The positive rates were 82.86%, 51.24% and 45.45%. Fifteen patients after EBUS-TBNA had a positive rate of 91.67%.

CONCLUSIONTBNA and EBUS-TBNA were proved to be safe procedure with a high yield for the diagnosis ofhilar and mediastinal lymph nodes in lung cancer patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biopsy, Fine-Needle ; methods ; Bronchi ; diagnostic imaging ; pathology ; Endosonography ; methods ; Female ; Humans ; Lung Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Lymph Nodes ; pathology ; Male ; Mediastinum ; pathology ; Middle Aged

Objective:To investigate the development and maturation process of intestinal organoids in neonatal mice so as to provide a new model for research on perinatal/neonatal intestinal epithelial development and related diseases.Methods:Intestinal tissue of 3-day-old C57BL/6 mice were collected and cultured for mouse intestinal organoids (MIOs) under standard conditions down to the fifth generation. The morphological changes of MIOs were observed and recorded using inverted phase contrast microscope. Real-time fluorescent quantitative polymerase chain reaction and immunofluorescence technique were used to detect the expression and location of markers of intestinal stem cells and differentiated cells of intestinal epithelium among different generations of MIOs (Selected marker genes: Lgr5 for intestinal stem cells, Tpm2 and Gja1 for fetal intestinal progenitor cells, Villin for intestinal epithelial cells, Lyz1 for Paneth cells, Muc2 for goblet cells, Chga for endocrine cells; Selected marker proteins: villin for intestinal epithelial cells, mucin 2 for goblet cells, chromaffin A for endocrine cells, lysozyme for Paneth cells). One-way analysis of variance and Bonferroni test were adopted for statistical analysis. Results:Two types of MIOs were observed, immature spheroid and mature organoids with crypt-villus structure. Spheroid was the main form in the primary culture. From primary to the second generation, the proportion of spheroids decreased from (96.61±1.36)% to (8.93±1.50)%, and so did the size ( F=12.88, P<0.001). During the second to the fifth generation, mature organoid, as the main form, increased from (91.07±1.50)% to (95.56±2.14)%. The expression of intestinal stem cell marker Lgr5 in the second generation decreased to 0.40±0.06 times of the primary one ( F=76.75, P<0.001) and then increased after this period. The expression of fetal intestinal progenitor markers Tpm2 decreased significantly during the passage (primary generation: 1.00±0.11, the fifth generation: 0.003±0.001, F=148.00, P<0.001); And the expression of Gja1 decreased from primary generation (1.00±0.14) to the second generation (0.06±0.04) ( F=197.10, P<0.001), but kept stable from the second to fifth genetation ( F=2.20, P=0.13). The expressions of gene markers of differentiated cells in intestinal epithelium, including enterocytes, goblet cells, endocrine cells, and Paneth cells, increased after the second generation (the second generation: Villin: 0.46±0.11; Muc2: 0.68±0.29; Chga: 2.53±0.16; Lyz1: 0.98±0.21; the fifth generation: Villin: 1.02±0.05; Muc2: 8.79±0.61; Chga: 4.32±0.45; Lyz1:3.81±0.36; all P<0.05). Immunofluorescence showed that villin, the intestinal epithelial cell marker protein, was distributed along the villus-side of MIOs in primary and the fifth generation culture. Mucin 2 from goblet cell and chromaffin A from endocrine cell expressed at a very low level in the primary generation, while higher in the fifth generation. In the primary culture, lysozyme from Paneth cell was evenly distributed in organoid cells, and high fluorescent dot-shaped expression was observed in the fifth generation. Conclusions:The development and maturation of immature intestinal epithelium can be simulated by continuous culture of neonatal MIOs. MIOs between the primary and second generation could be used as a research model for development of perinatal intestinal epithelium, and the second to the fifth generation as a model for neonatal intestinal diseases studies.