To study the cartilage differentiation of mouse mesenchymal stem cells (MSCs) induced by cartilage-derived morphogenetic proteins-2 in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2 (0, 10, 20, 50 and 100 ng/mL). After 14 days of induction, morphology of cells was observed under phase-contrast microscope. Collagen II mRNA and protein were examined with RT-PCR, Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining. RT-PCR showed that CDMP-2 could promote expression of collagen II mRNA in an dose-dependant manner, especially at the concentration of 50 ng/mL and 100 ng/mL. Immunocytochemistry and Western blotting revealed a similar change. Alcian blue staining exhibited deposition of typical cartilage extracellular matrix. Our results suggest that mouse bone marrow mesenchymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro, which provides a basis for further research on the role of CDMP-2 in chondrogenesis.
Bone Marrow Cells/*cytology ; Bone Morphogenetic Proteins/*pharmacology ; Cell Differentiation/*drug effects ; Cells, Cultured ; Chondrocytes/*cytology ; Chondrogenesis/drug effects ; Chondrogenesis/physiology ; Mesenchymal Stem Cells/*cytology ; Recombinant Proteins/pharmacology

Objective In clinical practice,ultrasound-guided fine-needle aspiration biopsy (US-G FNAB) is the gold standard in diagnosing the pathological nature of undetermined thyroid nodules before operation.Re---sults of FNAB may vary substantially among operators with different skills.The aim of this study is to evaluate whether operator experience in US-G FNAB influences the rate of diagnostic procedures.Methods A total of 600 consecutive US-guided FNAB done by a single radiologist with 22-or 27-gauge needles from Feb.2008 to Apr.2012 were retrospectively analyzed.All specimens were prepared and fixed without the cytologist on site and were subsequently analyzed by 2 expert cytologists.The procedures were chronologically divided into 6 groups and further stratified according to the overall (A),solid nodules (B),mixed nodules (C),pure cystic nodules (D),acellular sample(E),heavily blood-stained sample(F),exclusively colloid material(G),incorrect slide fixation(H) and they were classified as diagnostic or no-diagnostic.Results The rate of no-diagnostic procedures for each group in learning curves were reported as:① Among A,34％ in group 1,15％ in group 2,18％in group 3,10％ in group 4,7％ in group 5,and 8％ in group 6.② Among B,4％ in group 1,1％ in group 2,0％ in group 3,0％ in group 4,0％ in group 5,and 0％ in group 6.② Among C,10％ in group 1,1％ in group 2,4％ in group 3,0％ in group 4,1％ in group 5,and 0％ in group 6.④ Among D,20％ in group 1,13％ in group 2,14％ in group 3,10％ in group 4,6％ in group 5,and 8％ in group 6.⑤ Among E,25％ in group 1,15％ in group 2,15％ in group 3,10％ in group 4,6％ in group 5,and 8％ in group 6.⑥ Among F,4％ in group 1,0％ in group 2,1％ in group 3,0％ in group 4,0％ in group 5,and 0％ in group 6.⑦ Among G,1％ in group 1,0％ in group 2,1％ in group 3,0％ in group 4,1％ in group 5,and 0％ in group 6.⑧ Among H,4％ in group 1,0％ in group 2,1％ in group 3,0％ in group 4,0％ in group 5,and 0％ in group 6.Among the 600 procedures,there were 2 cases of vertigo and 4 cases of moderate bleeding at the biopsy site,all of which resolved spontaneously.No major complications were recorded.Conclusions The rate of no-diagnostic US-guided FNAB is heavily dependent on the operator's experience.We estimate that at least 200 procedures are needed in order to achieve the levels of diagnostic accuracy reported in the literature.We therefore suggest operators to get specific training before routinely perform this procedure in clinical practice.

Objective To study the pattern of cerebral gray matter and white matter volume changes among smokers with differ-ent level of nicotine dependence (addition)using voxel-based morphometry (VBM).Methods The current case-control study recrui-ted 53 healthy male smokers and 53 healthy non-smokers from outpatients of our hospital during January 2013 to May 2014.Personal information (including for example age,sex and addition dependence of subjects)was collected using a questionnaire.3D-T1 images of whole brain structure were collected and were analyzed using DARTEL toolbox of SPM8.Smokers were divided into mild to mod-erate nicotine dependence group (n=23)and severe nicotine dependence group (n=30)based on Fagerstr?m Test for Nicotine De-pendence (FTND)score.Independent sample t-test analyses were performed to compare the volumes of gray matter and white mat-ter between smokers with different levels of nicotine dependence and non-smokers.Results Compared with non-smokers,gray and white matter volumes of smokers were smaller in multiple brain areas,mainly in the middle occipital gyrus,posterior cingulate,cer-ebellum anterior lobe,precuneus,caudate body and insula,which however,had larger number and scope of focal areas with gray and white matter atrophy in the mild to moderate nicotine dependence group than that in the severe nicotine dependence group.Conclusion Smokers with mild to moderate nicotine dependence have more pronounced gray and white matter atrophy than that smokers with severe nicotine dependence have.

Skin temperature is relative to blood circulation. The temperature of transplanted skin is one of the most important indexes describing blood circulation after operation. This paper introduces a blood circulation monitor for transplanted skin, which monitors blood circulation through continuous testing of temperature.

OBJECTIVE: To suppress NgR gene expression in neural stem cells and observe differentiation of neural stem cells in vitro after interfered which provide nutritional support for the facial nerve repair in vivo. METHOD: PCR amplification, restriction endonuclease digestion, T4DNA ligase connections were used to connected NgR with rector pGCsi, and constructed recombinant vector (NgR shRNA). Lipofectamine 2000 were used to transfect the NSC. The expression of NgR was examined by Western Blot. The proportion of neural stem cells transformed into neurons after transfection was tested by Immunocytochemistry. Neural stem cells were planted in PLGA tubes after transfected, and were scanned by electron microscopy. RESULT: NgR shRNA plasmid was constructed and infected neural stem cells successfully. Western Blot showed that the expression of NgR decreased in neural stem cells after interference. Immunocytochemistry showed that the rate of the neural stem cells transformed into neurons after interfered was significantly higher (P < 0.01). CONCLUSION Neural stem cells were transformed into neurons after NgR shRNA plasmid infected neural stem cells, which promoted axonal regeneration more effectively and provided a efficient and stable gene platform for facial nerve repair.
Animals ; Cell Differentiation ; Cells, Cultured ; Facial Nerve ; surgery ; GPI-Linked Proteins ; genetics ; metabolism ; Myelin Proteins ; genetics ; metabolism ; Neural Stem Cells ; cytology ; metabolism ; Nogo Receptor 1 ; RNA Interference ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; genetics ; metabolism

Objective To construct mouse model of acute myeloid leukemia and detect the expression of ANGPT 1 ,ANGPT2 and VEGF gene on the cells its as well as the clinical significance .Methods The HL‐60 cells were transfected to NOD/SCID mouse through abdominal injection to construct mouse model of acute myeloid leukemia .Then identify mouse model by histopathology and Flow Cytometry .The expression of ANGPT2 ,ANGPT1 and VEGF mRNA in the tumor tissues of mouse model was detected by real‐time fluorescence quantitative PCR .The expression of ANGPT2 will be analyzed on the survival time of mouse model by Spearman′s correlation method .Results Mouse model has been successfully identified by histopathology and Flow Cytometry .The expression of ANGPT2 and VEGF in mouse mode was significantly detected ,which was that of higher than normal group (P<0 .05) .The expression of ANGPT1 was lower than that of ANGPT2 and VEGF ,there was no significant difference between AN‐GPT1 and normal groups (P>0 .05) .The higher expression of ANGPT2 in mouse model had a short survival time in mouse with acute myeloid leukemia .Conclusion This study showed that ANGPT2 mRNA was over‐expressed in acute myeloid leukemia .The increasing expression of ANGPT2 mRNA may lead to poor prognosis in mouse with acute myeloid leukemia .

Objective:To analyze the mechanism on integration of health care services at the county and town-ship level from the perspective of stakeholder theory. Methods: The stakeholder interest demand was determined by word frequency analysis of interview data from stakeholders in the three regions of Qianjiang, Huangpi, and Zhen-jiang;the degree of attention and gains and losses of stakeholders towards various demands was investigated from the three regions through interest demand questionnaires;the impact of demand benefits on behavioral responses has been evaluated through comprehensive evaluation theory and game theory. Results:Regional integration policies reflect the interest demands of stakeholders in varying degrees; the higher were the scores of demand benefits in interest de-mands questionnaires, the stronger was the willingness of stakeholders to coordinate integration policies. Conclusion:The policies of integration of health care services in rural China should consider all stakeholder interest demands;the better the interest demands of the stakeholders are satisfied, the stronger their motivation for integration reform will be, which may affect the implementation effects of local integration reforms to some extent.

Objective To summarize the clinical features,diagnostic and treatment experience of severe hand,foot and mouth disease(HFMD) cases receiving mechanical ventilation from Jan 1 to Sep 6,2009 in our hospital and provide reference for reducing the occurrence of neurogenic pulmonary edema(NPE) and mortality. Methods 147 severe HFMD who received mechanical ventilation were analyzed by a retrospective investigation. Results 85.0% children were less than 3 years old and 100% patients had a fever. The mechanical ventilation occurred within 1 to 4 days after fever (3.06 ± 1. 02) d. Neurological complications presented as bad spirit,easy surprised, involuntary movement of the extremities and lethargy. Respiratory system complication presented as polypnea, hypepnea and irregular rhythm. Circulatory system complication presented as heart rate increase or decrease, hypertension or hypotension, piebald skin and low limb temperature.Some of the children had high white blood cell counting,glucose and lactic acid of the blood. The EV71-PCR positive rate was 34. 7% for throat swabs,and 42. 2% for anal swabs. Chest X-ray presented as increased broncho vascular shadows, exudation or nonapparent abnormality. Only three children were dead, fatality rate was 2. 0%. Conclusion The HFMD cases in this outbreak are relatively more serious,and change rapidly. It's difficult to treat as soon as NPE appeared,and it has high fatality rate. Early recognition of critical patients,timely giving mechanical ventilation,and the comprehensive treatment can reduce NPE incidence and the mortality.

OBJECTIVE:To establish a method for determining the plasma concentration of paeoniflorin and phillyrin and phar-macokinetic study before and after intragastric administration of Qianliean granules. METHODS:LC-MS/MS method was adopted. The column was Waters C18 with mobile phase consisted of acetonitrile(A)-2 mmol/L ammonium acetate(containing 0.05% formic acid)(B)(0-9 min:15%A→50%A;9-11 min:50%A→90%A;11-17 min:90%A;17-19 min:90%A→15%A;19-20 min:15%A),at the flow rate of 0.6 ml/min;column temperature was 35 ℃ and the volume was 20 μl;quantitative ions were paeoniflorin m/z 525.2 → m/z 449.0,phillyrin m/z 552.3 → m/z 355.3. 7 SD male rats were docked to collect blood 0.5 ml from angular vein 0.25,0.5,0.75,1,1.5,2,3,4,6,8,10,12,24 h after administration Qianliean granule solution 1 g(medicinal materials)/kg to determine the blood concentration of drugs. DAS 2.1.1 software was employed to calculate pharmacokinetic parameters. RE-SULTS:The linear range of paeoniflorin and phillyri were 5.0-2500.0 μg/L(r=0.9979)and 2.0-2000.0 μg/L(r=0.9982),re-spectively;RSD of precision test was less than 5.5%(n=5);the method recovery were 96.0%-104.0% and 92.0%-107.0%,the extration recovery were 71.4%-83.5% and 81.5%-92.3% and RSD of stability test was less than 5.0%(n=3). The pharmacokinet-ic parameters of paeoniflorin and phillyrin were as follows as t1/2 of (2.206 ± 0.631) and (1.355 ± 0.317) h;cmax of (1504.069 ± 620.885) and (79.043 ± 15.568)μg/L;tmax of (1.000 ± 0.250) and (1.214 ± 0.267) h;AUC0-24 h of (4897.645 ± 2207.577) and (263.475±54.795)μg·h/L;CL of(5.025±2.773)and(76.253±13.986)L/(h·kg). CONCLUSIONS:The method is highly sensi-tive,exclusive,simple,accurate and reliable,and can be applied to study the pharmacokinetic characteristics of paeoniflorin and phillyrin in rats in vivo.

Objective To investigate clinical effect and safety difference of vinorelbine and docetaxel sepa-rately combined with lobaplatin in treatment of recurrent and metastatic breast cancer.Methods 160 patients with recurrent and metastatic breast cancer were chosen from July 2009 to July 2012 in our hospital,and they were random-ly divided into two groups,including A group (80 patients)with vinorelbine and B group (80 patients)with docetaxel on the basis of lobaplatin.The clinical efficacy for short -term,survival rate with follow -up and side effects incidence of both groups were compared.Results The RR and DCR of A group were 51.25%,68.75%,which of B group were 55.00%,71.25%.There was no significant difference in the clinical efficacy for short -term between the two groups (χ2 =1.04,2.37,all P >0.05).The survival rates in 1,2 and 3 years after treatment of A group were 60.00%,53.75%,28.75%,those of B group were 67.50%,57.50%,31.25%.There was no significant difference in the survival rate with follow -up between the two groups (χ2 =2.14,3.01,1.87,all P >0.05).The incidence rate of drug side effects of B group was significantly lower than A group(χ2 =13.14,9.33,15.74,11.65,8.29,all P <0.05).Conclusion Vinorelbine and docetaxel separately combined with lobaplatin in treatment of recurrent and metastatic breast cancer can efficiently control the disease progress and prolong the survival time;but docetaxel com-bined with lobaplatin treatment is helpful to reduce the risk of serious side effects.