Objective To develop and optimize a module for the automatic production of N-succinimidyl-4-[18F] fluorobenzoate (18F-SFB) that is used for further 18F labelling C2A domain of Synaptotagmin Ⅰ . The conjugated compound was applied for detecting the tumor apoptosis in rabbit model after chemotherapy. Methods GE TRACERlab and TRACERlab FXF-N modules were modified and programmed to automatically produce 18 F-SFB which was further analyzed by high performance liquid chromatography (HPLC).C2A-glutathione S transferase (GST) was conjugated with 18F-SFB (18F-FB-C2A-GST) and subsequently purified by HPLC. Two rabbits grafted with VX2 lung cancer were first treated with chemotherapy and then,37 MBq of 18F-FB-C2A-GST was administered via the auricular vein. Serial PET/CT imagings were performed at 0.5, 1 and 2 h post-injection respectively. Tumor apoptosis was examined by pathological study. Results The TRACERlab FXFoG and TRACERlab FXF-N modules were successfully adapted to synthesize18F-SFB, with the radiochenmical yield (76.41 ±4.00)% (n = 10), the corrected yield (45.43 ±5.90 ) % and the radiochemical purity about 95%. The whole procedure for labeling 18 F-SFB was about 87 min.From PET/CT imagings, significant uptake was found in the tumor after chemotherapy, but no obvious up-take was found in heart, lungs and liver. HE staining demonstrated large number of apoptotic bodies within the tumor tissues. Conclusions 18 F-SFB can be automatically synthesized. 18F-FB-C2A-GST might be useful for the detection of apoptosis in tumor after chemotherapy.

CD40/CD40L interactions play a pivotal role in T cell activation, and take part in many physiologic and pathologic procedures and different levels. In this article, stable CHO transformants secreting human CD40-Ig fusion protein were established through transfection and selection with Lipofectamaine and G418, respectively. In order to obtain great valume of recombinant protein, big batch serum-free cultures of engineered CHO cells were performed in roller-bottle using CHO-II-SFM medium. After cultures, the cell-culture supernatants were harvested, concentrated through ultra-filtration, and finally purified by affinity choromatography with Protein G Sepharose Fast Flow. Human peripheral bloods were collected freshly and seperated with Ficoll, CFU-T was cultured in semi-solid culture system with peripheral blood mononuclear cells (PBMNC). Effect of human CD40-Ig fusion protein on the formation of CFU-T was observed in vitro. The results showed that the yield of human CD40-Ig fusion protein was 30 mg in total 3 liter CHO-II-SFM culture supernatant, and it supposed that the expression level of CD40-Ig in CHO cells was more than 10 micro g/ml. The purity of purified fusion protein is above 95%. Furthermore, compared with human IgG, human CD40-Ig fusion protein significantly inhibited the formation of CFU-T at dose 0.25, 1.0, 4.0, and 10 micro g/ml, it lays a good foundation to evaluate its potential functions in vivo.

OBJECTIVETo discuss mass spectrum characterization of five valepotriates including 'monoene' type (didrovaltrate), 'diene' type (valtrate, acevaltrate) and 'four-olefinic' type (baldrinal and homobaldrinal) by electrospray ionization tandem mass spectrometry (ESI-MS(n)).

METHODThis study was carried out on the basis of electrospray ionization tandem mass spectrometric method and analysis of multistage fragments.

RESULTThe fragmentation patterns and structural assignment of 'monoene' type, 'diene' type and 'four-olefinic' type valepotriates in ESI-MSn under positive mode were summarized.

CONCLUSIONThe compounds have a strong pyrolysis rules and it can provide reference date for valepotriates in rapid structural identification, quantitative analysis and pharmacokinetic study.

Drugs, Chinese Herbal ; chemistry ; Iridoids ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

Embryonic hematopoiesis in mammals is characterized by successive temporal and spatial changes. Previous investigations indicate that in vitro differentiation of embryonic stem cells (ES cells) derived from 129 mice can mimic embryonic hematopoiesis to some extent. To investigate the in vitro hematopoietic differentiation capacity of ES cells derived from C57BL/6 mice, the authors initially established the murine ES cell line with standard identification methods employed. Next, two-step culture system was utilized for embryoid bodies formation and the appearance of different hematopoietic precursors was confirmed by CFC assay, cellular chemical staining as well as RT-PCR. The results demonstrated that the ES cell line MES-1 fulfilled the criteria of ES cell line and its progeny after in vitro differentiation included primitive and definitive erythrocyte precursors, mixed colony-forming cells and granulocyte/macrophage colony-forming cells. RT-PCR analysis revealed the molecular consistence of transcription factors and hematopoietic markers with cellular event. In conclusion, MES-1 established from C57BL/6 mice was able to differentiate in vitro to a variety of hematopoietic precursors, thus could partly recapitulate embryonic hematopoiesis.
Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; genetics ; Cell Line ; Colony-Forming Units Assay ; DNA-Binding Proteins ; genetics ; Embryo, Mammalian ; cytology ; Erythroblasts ; cytology ; metabolism ; Erythroid Precursor Cells ; cytology ; metabolism ; Erythroid-Specific DNA-Binding Factors ; Gene Expression ; Hematopoietic Stem Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism ; Time Factors ; Transcription Factors ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics

Recent reports have clearly demonstrated that bone marrow cells can be differentiated into neurons, suggesting the existence of cells with the differentiation capacity in the bone marrow cell population. It is well known that hematopoietic stem cells as well as mesenchymal stem cells (MSCs) can be transplanted and therefore, alternative of them might contribute to the process. In the present study it was addressed whether marrow MSCs could be coaxed into neuron-specific antigen bearing cells and if so, whether the differentiated cells possess the cytochemical features seen in neurons. The report here showed that high concentration of 2-mercaptoethanol (2-ME) could induce some of the MSCs into neuron-like cells expressing neurofilament (NF) and neuron specific enolase (NSE). The neuron-like cells were alkaline phosphotase positive while the others MSCs were kept negative. Cells treated with 2-ME were positive for alpha-naphthylacetate esterase and glycogen and negative for acetylchonlinesterase, which were similar with the results seen in untreated cells. Furthermore, Nissel body was not observed in treated cells shown by toluidine blue staining. Therefore, it is likely that the cells described here seem not belong to the neuronal lineage. These findings, however, reveal that human MSCs could alter their committed fates under some circumstances.

Objective To find out the perception status of Kaschin-Beck disease(KBD)-related knowledge among residents in Aba KBD areas. Methods In 2009, hierarchical clustering random sampling method was used to select 13 villages as survey points in Aba KBD areas, general demographic characteristics, KBD prevalence and KBD-related knowledge of residents were investigated. Results Of the residents investigated, the understanding rate of KBD-related knowledge was 36.7% (7361/20 080), understanding rate among female [40.2% (4427/11012) ]was high than that of male[32.3%(2934/9084), x2 = 134.80, P ＜ 0.05];50-year group[42.5%(2789/6562] was higher than others;Tibetan [42.8% (6775/15829)] was higher than other nationals;residents in Semi-agricultural and semi-pastoral areas [47.2% (5777/12239)] was higher than people in other areas ;farmer [42.6% (4585/10762) ],people who lost labor ability [42.7% (1487/3482)] and the unemployed [42.8% (941/2199) ] was higher;married people[41.6%(6067/14584)] was higher;KBD patients[47.6%(4585/9632)] was higher[x2 = 92.41,148.04,578.56,116.35,36.96,371.29 respectively, all P ＜ 0.05]. Sixty three point nine persent (978/1530) acquired KBD knowledge through explaination by medical and health personnel. Conclusions The current situation of perception of KBD-related knowledge among residents in Aba KBD areas is not optimistic. Understanding rate among residents with different demographic characteristics is significantly different. Targeted health education strategies and measures should be developed among different population groups.

OBJECTIVETo analysis the clinic and genotype in two Chinese patients with Dyskeratosis congenita (DC).

METHODSThe two patients were characterized by mucocutaneous abnormalities (abnormal nails, lacey reticular pigmentation, and oral leukoplakia), bone marrow failure. They were diagnosed with DC. DC genes were amplified by polymerase chain reaction (PCR), including DKC1, TERT, TERC, TINF2, NOP10, NHP2, then DNA sequencing was performed for abnormal exons.

RESULTSAn abnormal peak was found in exon 6 of TINF2 gene of the two patients. DNA sequencing showed a 845G→A transition in TINF2 gene in the two patients.

CONCLUSIONWe should think about DC if the young patients with mucocutaneous abnormalities and marrow failure. TINF2 c.845G→A(R282H) does exist in the two patients. It is reported in China for the first time.

Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Dyskeratosis Congenita ; diagnosis ; genetics ; Exons ; Female ; Humans ; Infant ; Male ; Telomere-Binding Proteins ; genetics

OBJECTIVEThis study was to identify the post-traumatic stress disorder (PTSD) changes and the relative risk factors within one year after Wenchuan earthquake among middle school students in the disaster area.

METHODSA total of 1966 students from 3 schools in Wenchuan earthquake region were selected as the target population. For each student, personal basic information and standard psychological scale (PCL-C, PSSS) were investigated by a self-administrated questionnaire in the 3rd, the 6th, the 9th and the 12th month after the earthquake, respectively. PTSD trends over the time and the associated risk factors were analyzed through the establishment of multi-level random coefficient model.

RESULTSThere were 1677 middle school students fully participated in the PTSD follow-up study by turning in the valid questionnaires. The averaged scores of PTSD at the time of the 3rd, the 6th, the 9th and the 12th month after the earthquake were 35.14 ± 11.08, 32.90 ± 11.03, 30.67 ± 11.28 and 29.75 ± 11.22, respectively. Meanwhile, the general incidences of PTSD were 36.6% (613/1677), 30.7% (515/1677), 24.8% (416/1677)and 22.2% (373/1677), respectively. The median score of perceived social support system was 60.00 and the general incidences of PSS was 17.20% (289/1677). The PTSD scores for the students had a decreasing trend during the period of our observation (β(time) = -1.879, χ(2) = 47.03, P < 0.05). The averaged scores for boys for the 4 follow-up studies were 33.71, 31.61, 29.66, 28.83; for girls were 36.33, 33.98, 31.51, 30.52; for junior school students were 35.46, 33.28, 30.18, 29.22; for senior school students were 34.89, 32.62, 31.04, 30.15. Moreover, two factors, gender and grade, were related with the decreasing trend (the trend for girls and senior school students was sharper than that for boys and junior school students) (β(gender-time) = -0.354, χ(2) = 4.83, P < 0.05; β(grade-time) = 0.622, χ(2) = 11.30, P < 0.05).

CONCLUSIONSThe prevalence of PTSD was high. Meanwhile, there was a trend of self-recovery for adolescent's PTSD during the first year of post-earthquake, but boys and junior school students recovered more slowly.

Adolescent ; Child ; China ; epidemiology ; Disasters ; Earthquakes ; Female ; Follow-Up Studies ; Humans ; Incidence ; Life Change Events ; Male ; Stress Disorders, Post-Traumatic ; epidemiology ; psychology ; Students ; psychology ; Surveys and Questionnaires ; Young Adult

Objective To investigate the level of serum superoxide dismutase(SOD),high-sensitive C-reactive protein (hsCRP)and myocardial injury markers in acute coronary syndrome(ACS)and evaluate their relationship and diagnostic values.Methods Case control study was conducted.This study enrolled 64 ACS patients and 50 non-ACS patients from the 181st Hospital of Chinese PLA in 2015.Serum SOD,hsCRP were tested and myocardial injury markers such as cardiac tro-ponin I(cTnI),creatin kinase MB(CK-MB)were also tested.Student t test and Pearson test were used as statistical meth-ods.Results Compared with control group,SOD of ACS group were significant lower(t=4.136,P<0.001)and hsCRP, Mb,cTnI,CK and CK-MB were significant higher(t=-5.396,-3.495,-5.578,-4.655 and -4.614,all P<0.001). The area under ROC curve of SOD,hsCRP,MYO,cTnI,CK and CK-MB was 0.713,0.758,0.699,0.879,0.841 and 0.802 respectively.After pearman test,the serum SOD were correlated to hsCRP,total cholesterol(TC)and low density lipopro-tein(LDL-C)(r=-0.493,0.548 and 0.404,all P<0.01).Serum hsCRP was correlated to cTnI,triglyceride(TG)and SOD(r=0.671,-0.417 and -0.493,all P<0.01).Conclusion cTnI was positively correlated to hsCRP and markers of myocardial injury.hsCRP was negatively correlated with SOD.It implicated that oxidative stress,inflammatory response and serum lipid deposition may act an important role in the occurrence and development of ACS.

Stem cell growth factor (SCGF) is an early-acting hematopoitic cytokine that has two isoforms including hSCGF with full length molecules and hSCGFbeta, 78 amino acids of which lost in the conserved calcium-dependent carbohydrate-recognition domain (CRD). It has been demonstrated that hSCGFbeta is strictly species-specific in regulating he-matopoiesis. This study was aimed to explore whether human SCGF can exert synergistic stimulatory effect on heterogenous murine CFU-GM progenitor. Firstly, hSCGF cDNA was amplified from human fetal liver cDNA library by using two-step PCR. The hSCGF mature peptide coding sequence was subsequently placed at downstream of glutathione S-transferase (GST) sequence in GST gene fusion expression vector. The results indicated that there existed an additional 60 kD protein compared with mock BL21 when the cells hosting recombinant plasmid were induced with IPTG at 37 degrees C. SDS-PAGE analysis demonstrated that the GST-hSCGF fusion protein mainly existed in insoluble form. When induced at low temperature (28 degrees C), the recombinant protein was mostly soluble. The GST-fusion recombinant protein was subsequently purified by using affinity chromatography. The clonogenic assay revealed that, unlike hSCGFbeta, hSCGF had the granulocyte/macrophage promoting activity (GPA) for murine bone marrow GM progenitor. It is concluded that, in contrast to human SCGFbeta, the intact molecular hSCGF may have no species specificity, implying that CRD domain in human SCGFbeta does not directly bind to corresponding SCGF receptor, but may have certain biological function.
Cloning, Molecular ; DNA, Complementary ; biosynthesis ; genetics ; isolation & purification ; Hematopoiesis ; genetics ; Humans ; Species Specificity ; Stem Cell Factor ; biosynthesis ; genetics ; isolation & purification