Objective To observe the effects of different collagenase digestions on isolating human umbilical cord mesenchymal stromal cells (MSC) from Wharton’s jelly, to exam their differentiation ability and to investigate their passage effect on the immune phenotype. Methods Human umbilical cord samples were digested by collagenaseⅠorⅡorⅣfor 4-18 hours then were passed through sieves . Cells were collected by centrifugation then inoculated in DMEM/F12 medium at concentration within range of 4.8×103-1×104/cm2 to compare the effect of different digestions on MSC. Von kossa staining and tetracycline fluorescence was used to label the osteogenic differentiation capacity of MSC. Also RT-PCR was employed to identify the differentiate capacity of MSC into myocardial-like cells. The immunophenotype of MSCs were detected by flow cytometry after subculture. Results Using collagenaseⅠdigestion, the number of MSCs isolated from human umbilical cord in Wharton’s jelly and their vitality were much higher while the period to show cell extension and primary culture time were shorter than those using collagenaseⅡorⅣdigestions. The analysis of surface marker revealed that the expression of positive markers include CD29, CD44, CD73, CD90 and CD105 did not change with passages while the negative markers such as CD31, CD34 and HLA-DR increased significantly with passages;Differential experiments induced in vitro show that human umbilical cord MSC in wharton’s jelly had the ability to differentiate into osteoblasts and myocardial-like cells. Con?clusion The human umbilical cord MSC in Wharton’s jelly was successfully isolated by collagenaseⅠdigestion. This meth?od was simple with a high success rate while cell loss and damage were minimum. This makes large-scale cultivation possi? ble. Negative markers increased with cell passages. This phenomenon revealed that MSC showed directional differentiation.

Objective: The purpose of this tudy was to dynamicly observe the weight and the bone mineral density (BMD) of the unsexed rabbits with a few self-made standardized phantoms. Methods:The eighteen healthy adult female rabbits were measured for their weight and BMD in preunsexed and postunsexed 5 months, 10 months with quantitative CT(QCT). Results:There were 61.1% of rabbits whose weight and BMD decreased after 5 months of the postunsexed and 100% of rabbits whose weight and BMD decreased after 10 months of the operation. Conclusion:QCT can be used to dynamicly observe curative effect of drugs in various periods as well,and it is a good method to study osteoporosis.

10.3969/j.issn.2095-4344.2013.23.004

BACKGROUND: In vitro culture condition and culture efficiency are different in reported umbilical cord-derived mesenchymal stem cells, and lacked of unified standards. Different derived mesenchymal stem cells have different biological properties. Therefore, it is very necessary to establish a simple and high-performance culture system for umbilical cord-derived mesenchymal stem cells. OBJECTIVE: To observe the growth state of human umbilical cord-derived mesenchymal stem cells in different culture systems in vitro and adenovirus infection efficiency. METHODS: Mesenchymal stem cells were separated from healthy full-termed delivery fetus using collagenase digestion method and purified by adherent culture. These cells were cultured and amplified in DMEM (low glucose), MesenPRO RS~(TM) Medium and STEMPRO~(R) MSC SFM in vitro. The 3-5 passage mesenchymal stem cells were infected by the Ad5-EGFP, Ad5/11-EGFP, Ad5/35-EGFP as multiplicity of infection (MOI)=1, 10, 100. Viral infection and green fluorescence expression were observed at post-infection 24, 56 and 72 hours using inverted fluorescence microscope. RESULTS AND CONCLUSION: The cell morphology in STEMPR~(R) MSC SFM was different from other two culture system and these cells were not easy to adherent after trypsin digestion. Cell doubling time in the MesenPRO RS~(TM) Medium was shorter than other two groups. Mesenchymal stem cells were infected by Ad5/35-EGFP with higher efficiency than other two kinds of adenovirus, but part of cells appeared apoptosis. The infection efficiency of Ad5/11-EGFP was highest. The fluorescence intensity was gradually increased with increased MOI.

Objective To assess the prevalence of Personality Diagnostic Questionnaire (PDQ)personality deviations in patients referred for functional dyspepsia (FD) with reliable and universal psychological measures, and to explore the relationship between co-occurring PDQ-personality deviations and functional dyspepsia. Methods The sample comprised 246 patients referred for functional dyspepsia. Four groups were divided according to their patterns of gastrointestinal symptoms: the FD group, FD with refluxlike symptom group(FD + RS group), FD with irritable bowel syndrome group( FD + IBS group), and FD with reflux-like symptom and irritable bowel syndrome group ( FD + RS + IBS group). Participants were assessed with the Personality Diagnostic Questionnaire for DSM-Ⅳ ( PDQ-4 ) to evaluate the presence of personality deviations. Results Overall 65% patients scored positive for any personality deviation, male and female alike. Cluster C (anxious/fearful) personality was most commonly found in FD patients (142 patients, 57.7% ). The FD + IBS group and the FD + RS +IBS group had significantly higher total PDQ scores than the FD group (23.39 ± 8. 77 and 24. 22 ± 10. 97 vs 18.98 ± 11.88, P ＜ 0. 05, respectively),indicating that FD patients with greater level of personality deviations tend to report other symptoms involving the esophagus and lower gastrointestinal tract. Reflux-like symptom without actual pathological acid regurgitation indicated cluster A (odd/eccentric) personality deviations. Conclusions The current study shows personality deviations are common in patients referred for functional dyspepsia. Negative emotions,maladaptive coping, and lack of social support, may strongly influence their healthcare-seeking behavior.There is no single personality type specific for some kind of gastrointestinal symptom. But FD patients with personality deviations tend to report other symptoms involving the esophagus and lower gastrointestinal tract.

BACKGROUND:High-quality and efficient umbilical cord-derived mesenchymal stem cells could be obtained by establishing and improving the method to avoid fungal contamination and by effectively decreasing pol ution rate of isolated culture during umbilical cord col ection. OBJECTIVE:To explore the effects of amphotericin B on growth and differentiation potential of umbilical cord-derived mesenchymal stem cells. METHODS:Umbilical cord-derived mesenchymal stem cells were separated from healthy ful-termed delivery fetus using col agenase digestion method and treated with amphotericin B. Subsequently, umbilical cord-derived mesenchymal stem cells were amplified and cultured in vitro with MesenPRO RSTM medium. The third passage of umbilical cord-derived mesenchymal stem cells in logarithmic phase was obtained to analyze their morphology, proliferation and immunophenotype, and induced to differentiate into osteoblasts and adipocytes in vitro. RESULTS AND CONCLUSION:After treatment with amphotericin B, umbilical cord-derived mesenchymal stem cells were successful y isolated and cultured in vitro. Flow cytometry results revealed that human umbilical cord-derived mesenchymal stem cells strongly expressed CD44, CD105 and CD73, CD90, and negatively expressed HLA-DR, CD29, CD31, and CD34. Amphotericin B-treated human umbilical cord-derived mesenchymal stem cells can stil differentiate into adipocytes and osteoblasts in vitro.

Objective To investigate the dynamic expression of the drug resistance protein P-glycoprotein (P-gp) within 72 hours in the pentylenetetrazol (PTZ)-induced status epilepticus (SE) model, and to identify the optimal detection time to inhibit P-gp. Methods mRNA and protein expressions of P-gp in rats hippocampal tissue were detected by using immunohistochemistry , RT-qPCR and Western blot at different time points after modeling (0, 3, 6, 12, 24, 48, 72 h). Results The mean density of P-gp protein in the hippocampus of status epilepticus model was 0.325 1 ± 0.008 2 at 24 h, and was 0.396 3 ± 0.016 8 at 48 h, which were consistently higher than those of the control group (P < 0.05, P < 0.01, respectively). Results of qRT-PCR showed that MDR1a expression was significantly upregulated at 24 h and at 48 h (P < 0.05, P < 0.01, respectively). Western blot assay revealed that P-gp protein was also significantly increased at 48 h after seizures (P < 0.05). Conclusions The upregulation of P-gp after SE peaked at 48 h, which maybe the optimal detection time to detect drug resistant after SE.

Early diagnosis of malignant lymphoma is often difficult since the clinical manifestation of the lymphoma occurred in the maxillofacial region is very similar to that of the squamous cell carcinoma.When the pathological diagnosis is not clear,the surgeon is easy to misdiagnose and lead to mistreatment.A patient visited the Affiliated Haikou Hospital of Xiangya School of Medicine,Central South Universily for gingival mass with an ipsilateral submaxillary enlargement.The clinical manifestation and preoperative MRI are very prone to squamous cell carcinoma with metastasis,so we did not take a preoperative pathological examination.The gingival mass was surgical removed firstly,but frozen pathological result showed that it was malignant small round cell tumor.Since the patient was diagnosed as high degree malignant of small round cell tumor and the submandibular region have been significantly metastasized,so we carried out the combined radical dissection of gingival,mandible and neck surgery.The postoperative pathological report was malignant lymphoma,suggesting that the patient was a case of misdiagnosis and mistreatment.This article draws lessons from misdiagnosis and provided experience for seeking improvement measures.

This paper discusses the theoretical basis of traditional Chinese medicine （TCM） for the treatment of cancer-related fatigue （CRF） based on the principle of treating overstrain with warming in the Huangdi's Internal Classic 《黄帝内经》 and the role of hypothalamic-pituitary-adrenal （HPA） axis dysregulation in CRF， aiming to provide new ideas for the clinical treatment of CRF. In the pulsatile regulation of the HPA axis， cortisol， which is synthesized and released by the adrenal cortex， plays an essential role in the life activities. The abnormal circadian rhythm and reduced serum level of cortisol are major factors leading to CRF. Therefore， increasing the serum level and stimulating the biological activity of cortisol and restoring the normal function of HPA axis are important targets for the treatment of CRF and also the key to the TCM treatment of this disease. According to the TCM principle of treating overstrain with warming， we interpreted the etiology and clinical manifestations of overstrain and further explored the causes of CRF. It is believed that the depletion of Qi and blood， Yin and Yang， and fluid in the kidney， spleen， and liver is the key of the disease， which results in symptoms such as fatigue. Further， we elaborated on the theoretical connotation of warming and summarized the two main treatment principles of tonifying with warm-natured herbs and relieving fever with sweet- and warm-natured herbs. According to the different characteristics of the organs affected by pathogen， we proposed the treatment method of warming kidney to cultivate essence， warming stomach and nourishing spleen to replenish Qi， nourishing liver and tonifying blood to promote Qi movement， and relieving fever with sweet- and warm-natured herbs to alleviate fatigue. The warming method can elevate the serum level and invigorate the biological functions of cortisol in CRF patients， which provides a new theoretical basis for alleviating the symptoms and improving the quality of life of cancer patients in clinical practice.

Circulating tumor cells(CTCs)are cells that dissociate from the tumor tissue and enter the lymphatic system or bloodstream with close association with tumor metastasis and recurrence.CTCs contain complete pathological information,which can be extracted by isolation,enrichment,and analysis of the CTCs to guide cancer diagnosis and treatment,thereby significantly improving the monitoring efficiency and prognosis of cancer.Compared with tissue biopsy,liquid biopsy with CTCs as a biomarker enables specific and dynamic detection of tumor growth with a less painful experience.For detection of CTCs,the cells must be captured from body fluids,followed then by their release and enrichment.This review summaries the latest research progress in responsive isolation of CTCs(e.g.with light,dielectrophoresis,acoustophoresis and magnetophoresis),chemical isolation(specific molecules and topological structure)and responsive release(e.g.,light,electric,thermal,pH,enzyme responsiveness,and substrates break).Responsive isolation utilizes the differences in physical properties between CTCs and blood cells,while chemical isolation utilizes specific recognition mechanisms to capture the CTCs.These techniques result in low cell damage with a high specificity to facilitate further analysis.Currently,CTC detection has been applied for early diagnosis and prognostic assessment of multiple cancers including lung cancer,liver cancer,colorectal cancer,and prostate cancer.