BACKGROUND: There are no studies on differentiation of bone marrow mesenchymal stem cells (BMSCs) into neurons directly induced by neural cells. OBJECTIVE: To establish the co-culture system between BMSCs and neural cells in vitro, and to study the influence of neural cells on the differentiation of BMSCs into neuron in the co-culture system. DESIGN, TIME AND SETTING: The in vitro cytology control experiment was performed at the Central Laboratory of Hospital Affiliated to Medical College of Qingdao University from December 2006 to December 2007. MATERIALS: BMSCS were harvested from bone marrow of patients with diabetes meliuts, who underwent autologous stem cell transplantation at the Hospital Affiliated to Medical College of Qingdao University. Neural cells were collected from brain tissues of infants die of asphyxia during delivery. The third passage of neural cells was used in this study. Transwell double-deck culture dish was purchased from Corning Costar, with a pore diameter of less than 3.0 ?m. Cells could not traverse, but the medium could traverse. METHODS: Neural cells were incubated in Transwell double-deck culture dish at a density of 1?106 in the co-culture group. BMSCs were incubated in the upper layer in LG-DMEM medium for 4-5 days. BMSCs were incubated in both layers in the control group. MAIN OUTCOME MEASURES: The morphological changes of BMSCs were observed and the special markers of neurons cells in BMSCs were examined by immunofluorescence. RESULTS: BMSCs in the co-culture group grew slowly, showing radial processes and connected each other. Positive rate of neuron specific enolase was (32.7?11.5)%. BMSCs in the control group were flat and wide, and negative for neuron specific enolase. CONCLUSION: Microenvironment provided by neural cells promotes the differentiation of BMSCs into neurons.

BACKGROUND: There is no ideal method about adult bone marrow mesenchymal stem cells (BMSCs) differentiate into islet-secreting cells and appreciation in vitro at present. Transgene requests strict conditions and complex program. The induction using chemicals is a present-used method. OBJECTIVE: To investigate the culture conditions for inducing adult BMSCs into islet-like cells in vitro. DESIGN, TIME AND SETTING: The cytology in vitro experiment was performed at the Central Laboratory of Affiliated Hospital of Medical College of Qingdao University from January to October 2007. MATERIALS: Bone marrow was collected from diabetic patients undergoing autologous stem cell transplantation at Department of Endocrinology, Affiliated Hospital of Medical College of Qingdao University after signing the informed consent. Epidermal growth factor and basic fibroblast growth factor were obtained from Peprotech Asia. B27 adjunct was purchased from Gibco. Nicotinamide, 2-mercaptoethanol, glutamine and dithizone were bought from Sigma, USA. METHODS: The adult BMSCs were isolated by Percoll from adult bone marrow aspirates and cultured in LG-DMEM, were suspended by Trypsin and passaged for subsequent passages. At the third to fifth passages, BMSCs were incubated at a density of 1?108 L-1 and were induced differentiation into islet-like cells through three developmental stages. In the first stage, BMSCs were incubated in HG-DMEM supplemented with 2-mercaptoethanol, glutamine for 2 days. In the second stage, BMSCs were incubated in HG-DMEM supplemented with epidermal growth factor, basic fibroblast growth factor, B27 and glutamine for 6 days. In the third stage, BMSCs were incubated in HG-DMEM supplemented with nicotinamide and 2-mercaptoethanol for 6 days. BMSCS in the control group were only incubated in the HG-DMEM. MAIN OUTCOME MEASURES: Morphological changes in BMSCs were analyzed under a phase contrast microscopy. Duodenal Homeobox 1 (PDX-1) expression was detected during the second and differentiation stage by immunofluorescence assay. Islet ?-like cell clusters from the 3rd stage were identified by positive dithizone staining. The insulin secretions under the stimulation of high or low glucose were detected by electrochemiluminescence immunoassay. RESULTS: The undifferentiated BMSCs exhibited adherent long spindle-shaped cells. After induction, cells gradually became round and formed clusters. Cells expressed the PDX-1 gene at 8 days and formed islet-like cell clusters that exhibited positive dithizone staining at 14 days. After high and low glucose treatment, no insulin was detected in the control group; insulin content significantly increased at 14 days (t=3.638-9.387, P

Objective To explore the effectiveness of transcutaneous electrical acupoint stimulation (TEAS) approach in treating patients with cancer- related fatigue during periods of chemotherapy in non-small cell lung cancer (NSCLC) patients in China. Methods A total of 162 participants who treated with GP chemotherapy were randomly assigned to three groups: 66 cases in control group, 61 cases in TEAS group, 67 cases in Sham TEAS group. The following acupoints were used in this study: Qihai (CV 6), Keshu (UB 17), and Zusanli (ST 36). Participants in TEAS group and Sham TEAS group received eight 30-min sessions of TEAS over 28 days. The Revised Piper Fatigue Scale (RPFS) and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core 30 (EORTC-QLQ-C30) were used to measure cancer related fatigue (CRF) on the day before chemotherapy days 8 and 28 separately. The differences among three groups were analyzed. Results Finally, 167 patients were included in this study, 56 cases in control group, 57 cases in TEAS group, 49 cases in Sham TEAS group. At the 28th day, the outcomes of the RPFS for TEAS group, Sham TEAS group and control groupscored 2.06 ± 0.90, 2.80 ± 1.34, 3.00 ± 1.29 respectively. There were significantly different among three groups (F=9.784,P<0.01). At the 28th day, the outcomes of the EORTC-QLQ-C30 for TEAS group, Sham TEAS group and control groupscored 64.56 ± 5.00, 54.90 ± 6.25, 54.48 ± 9.68 respectively. There were significantly different among three groups (F=34.119, P<0.01). Conclusions TEAS could help to relived cancer-related fatigue.

A stimulate method for determination of polybrominated diphenyl ethers ( PBDEs) and derivatives (OH-PBDEs and MeO-PBDEs), tetrabromobisphenol A (TBBPA), hexabromocyclododecane (HBCD) in egg samples was developed by gel permeation chromatography ( GPC) and dispersive solid phase extraction ( DSPE) combined with liquid chromatography tandem mass spectrometric ( HPLC-MS/MS) and gas chroma-tography-negative chemical ionization mass spectrometry ( GC-NCI/MS ) . The analytes were extracted with mixture of hexane and dichloromethane (1∶1, V/V) by accelerated solvent extraction (ASE), and purified by 100 mg C18 dispersive solid phase extraction ( SPE) sorbents followed with gel permeation chromatography (GPC) , and then analyzed by liquid chromatography tandem mass spectrometric (HPLC-MS/MS) and gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS), respectively. The quantita-tion was carried out external standard method. The recoveries of objects were 64. 5%-97. 2% and 65. 6%-109 . 2% ( except BDE85 was 54 . 8%, OH-BDE-137 was 47 . 4%) spiked at 1 . 0 μg/kg or 5 . 0 μg/kg in egg white and egg yolk, respectively. The relative standard deviations (RSDs) were less than 20. 2%. The limits of quantitation (LOQ) for the object were 0. 01-0. 2 μg/kg.

Objective To analyze the clinical data, including clinical features, treatment, and prognosis,in patients with different kinds of pituitary adenomas. Methods In this retrospective study, 746 cases were included. The characteristics of general epidemiology, clinical symptoms, pathology, imaging, treatment, and prognosis were analyzed. Results Clinical features were different among various pituitary adenomas. Symptoms caused by mass effect and hormone abnormality were expresssed in varying degrees. Serum prolactin>121.28 μg/L can differentiate the prolactin adenoma from the other huge tumor causing hyperprolactinemia due to the mass effect. There is significant relationship between the size and the various types of pituitary adenomas ( P<0.01 ),also between the size and the invasive capability ( P<0.01 ). Conclusions The pituitary adenomas may have their specific epidemiological, clinical, pathological, and imaging features, due to the distinct biological behavior. It is necessary to do the diagnosis, treatment, and prognosis evaluation individually.

Objective To evaluate the prevalence of NI in the SICU at our hospital. Methods 181 NI patients in the SICU were retrospectively analysed during Jan 1996～Dec 2000.Results The average NI rate was 9.81%. The major sites of NI were respiratory tract(36.96 %),thoracic/abdominal cavity(25.47 %)and bloodstream infections(9.32 %).The difference in major pathogens of infections in different sites reached statistical significance. For respiratory tract, thoracic/abdominal cavity and bloodstream infections,bacteria were the most common pathogens. Fungi were the moat frequent isolate from urine and stool. Mixed infection proportion was 52.25 %. The most common pathogens were Enterococci, Methicillin resistant Staphylococci、 Pseudomonas Aecruginosa、Escherichia Coli、Candida Albicans and Candida Tropicalis. Conclusions The most common pathogens of NI in SICU are different in different infection sites. The pathogens were complicated and most strains are antibiotics resistant. So it is important to establish NI control and to understand the changes of pathogens so as to prevent the infection.

Codon bias is an important factor which influences heterologous gene expression. Optimizing codon sequence could improve expression level of heterologous gene. In order to improve the expression level of BmK AngM1 gene encoding the analgesic peptide from Buthus martensii Karsch in Pichia pastoris, the codon-optimized BmK AngM1 gene according to its cDNA sequence and the preference codon usage of P. pastoris were cloned into expression vector pPIC9K and then transformed into P. pastoris. The expersion of recombinant BmK AngM1 (rBmK AngM1) was inducced by methanol in the medium, and the expression level of the optimized BmK AngM1 gene was 3.7 times of the native one. These results suggested that the expression of BmK AngM1 in P. pastoris could be successfully improved by codon optimization.

Objective To study the changes of serum interleukin-8 (IL-8) and interleukin-10 (IL-10) in preterm infants delivered from mothers with chorioamnionitis (CA) and the possible effects on bronchopuhmonary dysplasia (BPD) and brain injury of preterm infants.Method From October 2014 to October 2015,clinical data from mothers without clinical manifestations of CA giving birth to a preterm baby (gestational age:26 to 33 weeks) were collected in Changzhou Maternal and Child Health Hospital and Suzhou Municipal Hospital.The infants were assigned to CA group and non-CA group according to their mother's placenta pathology.The levels of serum IL-8 and IL-10 were measured using enzyme-linked immunosorbent assay (ELISA) on day 1 and day 7,while the levels of WBC,CRP and PCT were measured at birth.The incidences of BPD and brain injury were also reviewed.Result A total of 67 preterm infants were included,with 51 in the CA group and 16 non-CA group.The levels of IL-8 were significantly higher in the CA group than the non-CA group on both day 1 and day 7 [(21.6 ±9.7) ng/L vs.(7.3 ±2.3) ng/L,(26.6 ± 12.9) ng/L vs.(7.3 ± 2.1) ng/L].The IL-10 levels were significantly lower on day 1 [(1.92±0.17) ng/Lvs.(2.04±0.18) ng/L] and higher on day 7 [(2.42±0.60) ng/L vs.(2.03 ±0.18) ng/L] in the CA group.Significant differences existed on the incidences of BPD (54.9％ vs.25.0％) and brain injury (74.5％ vs.43.8％) between the two groups (P ＜ 0.05).The levels of CRP and PCT were higher in CA group (P ＜ 0.05) and the WBC was similar between the two groups (P ＞ 0.05).In CA group,on both day 1 and day 7,infants with BPD had similar IL-8 and IL-10 levels comparing with infants without BPD(P ＞ 0.05),also were infants with brain injury comparing with infants without brain injury.Conclusion Chorioamnionitis in pregnant women may affect serum cytokines levels in premature infants and lead to high incidences of BPD and brain injury.

Objective:To investigate the clinical value of dynamic detection of lymphocyte subsets and blood cell counts in management of patients with lupus nephritis (LN).Methods:The clinical data of 65 patients with primary LN admitted in Affiliated Hospital of Qingdao University from January 2015 to April 2021 were retrospectively analyzed. According to the stage of disease progression and medications used,LN patients were classified into primary active phase,post-induction therapy phase,and maintenance therapy phase. The changes in lymphocyte subsets were monitored,and the relationship of lymphocyte subsets and blood cell count ratios with lupus activity and infection events was evaluated.Results:The decrease of CD4 +T lymphocyte and NK cell counts were negatively correlated with the activity of systemic lupus erythematosus (SLE)( r=-0.67,-0.33, P<0.01),while CD8 +T lymphocyte,B cell counts,neutrophil/lymphocyte ratio (NLR),platelet/lymphocyte ratio (PLR),and monocyte/lymphocyte ratio (MLR) were positively correlated with the SLE activity( r=0.38,0.26,0.34,0.26,0.29, P<0.05). The area under ROC curve (AUC) of CD4 +T lymphocyte count in predicting the occurrence of infection in LN patients was the highest (0.89); taking 247.50 cell/μl as cutoff value,the sensitivity and specificity were 81.25% and 87.50%,respectively. The combination of CD4 +T lymphocyte with CRP increased the predicting value for the occurrence of infection. Conclusion:Dynamic detection of blood lymphocyte subsets and blood cell counts can reflect SLE activity and the occurrence of infection in LN patients. Among these indicators the CD4 +T lymphocyte has the highest predictive value for the occurrence of infection,and the combination of the CD4 +T lymphocyte count with CRP level can further improve the predicting value.

OBJECTIVETo study Twist expression in thyroid papillary carcinoma (PTC) by immunohistochemistry and to assess its usefulness as marker in the differential diagnosis of PTC, follicular adenomas (FA) and benign papillary lesions (BPL).

METHODSFifty cases of PTC, 48 cases of FA and 47 cases of BPL were evaluated using manual tissue chip and SP immunohistochemical stain to detect the expression of Twist and HBME-1, and comparing the staining to that of cytokeratin 19 (CK19).

RESULTSIn PTC, positive rates of Twist, HBME-1 and CK19 were 100% (48/48), 94.0% (47/50) and 78.0% (39/ 50) respectively; in FA, positive rates were 0, 6.7% (3/45) and 0 respectively; in BPL, positive rates were 7.0% (3/34), 2.1% (1/47) and 0, respectively. The differences between PTC and FA and between PTC and BPL were both statistically significant (P = 0. 000). The sensitivity of Twist, HBME-1 and CK19 was 100%, 94.0% and 78.0%; the specifity was 96.4%, 95.7% and 100%; overall accurary was 97.7%, 95.1% and 91.9%, respectively.

CONCLUSIONSPositive rates of Twist is higher than the other markers in PTC. Immunohistochemical staining of Twist has important significance in the differential diagnosis of thyroid lesions. Twist immunohistochemistry maybe helpful in diagnosis and differential diagnosis of PTC.

Adenocarcinoma, Follicular ; metabolism ; Adenocarcinoma, Papillary ; pathology ; Adenoma ; diagnosis ; metabolism ; Biomarkers, Tumor ; immunology ; Carcinoma, Papillary ; diagnosis ; metabolism ; pathology ; Carcinoma, Papillary, Follicular ; metabolism ; Diagnosis, Differential ; Galectin 3 ; genetics ; metabolism ; Immunohistochemistry ; Keratin-19 ; genetics ; Keratins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Thyroid Neoplasms ; diagnosis ; metabolism ; pathology ; Thyroid Nodule ; pathology ; Twist-Related Protein 1 ; genetics ; metabolism