BACKGROUND: A few mesenchymal stem cells (MSCs) can be found in peripheral blood.OBJECTIVE: To isolate rabbit peripheral blood MSCs and induce its differentiation into osteoblasts.DESIGN, TIME AND SE'I-I'ING: The cytological in vitro study was performed at the Central Laboratory of Shangdong Provincial Hospital from June to December 2008.MATERIALS: A total of 6 New Zealand rabbits were purchased from Shandong Academy of Agricultural Sciences. α-MEM and L-DiEM were bought from Hyclone.METHODS: Full-thickness skin (3 cm×3 cm) (dorsal muscular layer was left) was incised at various sites of rabbit back, every 3days. Incised skin was dressed following orthotopic transplantation. Each rabbit received four consecutive wounds. Peripheral blood was collected from femoral vein before injury and 1 week after injury. MSCs were harvested from peripheral blood by density gradient cantrifugation. MSCs were divided into 2 groups, which were respectively incubated in α-MEM supplemented with 10% fetal bovine serum and L-DiEM. Cells at passage 2 and 1 ×105/cm2 were incubated in a 12-well plate and induced with H-DiEM containing osteogenic inductor.MAIN OUTCOME MEASURES: The following parameters were measured: cell morphology before and after injury; colony forming efficiency of MSCs; outcome of osteogenic induction.RESULTS: Following primary medium change and before injury, obtained cells were normal. Twenty-four hours following incubation and after injury, MSCs were spindle or polygonal, and adhered to the wall. 5-6 days later, cell colonies appeared.Compared with the L-DiEM group, the number of primary culture colony formation in α-MEM group was significantly greater (P < 0.05). Peripheral blood MSCs were spindle, tdangle or polygonal, with the presence of processes, 2-3 nuclei and cell division phase, and slowly proliferated following osteogenic induction. At day 7 after differentiation of MSCs into osteoblasts, positive rate of alkaline phosphates was above 80%. At day 21, calcium deposition was detected by Alizarin Red S (positive staining).CONCLUSION: MSCs could be harvested from peripheral blood of wounded rabbits, with characteristics of osteogenic differentiation, α -MEM was more suitable than L-DiEM for peripheral blood MSCs to growin vitro.

Objective To compare the analgesia effect and the safety of Flurbiprofen Axetil (FA) and Parecoxib Sodium (PS) after posterior lumbar fusion surgery. Methods 90 patients undergoing internal fixation of lumbar spine randomly assigned to 3 groups:those in Group A(n = 30) received 100 mg of FA; those in Group B (n=30) received 40 mg of PS and those in Group C received saline.The VAS scores of 2, 6, 12, 24, 48, 72 h after operation and the dose of tramadol hydrochloride (TH) used and the side effect was recorded respectively. Results Group A and B had significantly better analgesic effect than Group C(P<0.05). Group A and B had lower average dose of TH than Group C (P<0.05). The VAS scores in Group A was lower than that in Group B in 2 h after the surgery. The VAS scores after the surgery showed no significant difference between Group A and B in 6 , 12, 24 h after the surgery. The VAS scores in Group A was higer than that in Group B in 48,72 h after the surgery. Conclusion Both PS and FA can alleviate postoperative pain and have fewer adverse reactions.

Human gait involves a complex mechanism of muscular skeletal coordinated operation,which is specific and can be used as the basis of identity recognitions and clinical disease diagnoses.Human gaits have wide application value in the field of disaster rescue,battlefield ambulance,counter-terrorism,security,and medical and healthcare.The traditional contact-free gait detection technology mainly depends on optical images or ultrasound,which is susceptible to light,low visibility,obstacles,etc.In recent years,with the rapidly development of bio-radar technology,the bio-radar based contact-free human gait signal detection technology has shown more advantages.It can not be affected by light,can penetrate clothing,camouflage or even walls,and can operate in all-weathe,including low visibility weather conditions such as smog,smoke and fog.In this paper,the technical principles and methods of bio-radar based contact-free human gait detection technologies were discussed,the research status was summarized,and the development trendency was prospected.

Objective To explore the CT and pathologic manifestations of the pulmonary chondroma,to improve the diagnostic accuracy and reduce the misdiagnosis.Methods The CT data of 6 patients with pulmonary chondroma proved by pathology were analyzed retrospectively.Results All 6 cases were solitary,3 cases occurred in the right lung and the other 3 cases occurred in the left lung.The diameter of the lesions ranged from 1.0 cm to 5.1 cm.2 cases showed lobulated shape,4 cases showed round shape.4 cases showed circumscribed margin,2 cases showed blurrmed margin.Furthermore,calcification was detected in 1 case.Conclusion Pulmonary chondroma has some characteristic CT features,including vascular border sign and begonia sign.However,it should be differentiated from pulmonary hamartoma,peripheral lung cancer and sclerosing pneumocytoma.

The aim of this study was to establish a high performance liquid chromatography-mass spectrometry method for the determination of 5-(4′-(bromomethyl)-［1, 1′-biphenyl］-2-yl)- 1H-tetrazole(BBT1)and 5-(4′-(dibromomethyl)-［1, 1′-biphenyl］-2-yl)-1H-tetrazole(BBT2), which are two genotoxic impurities in olmesartan medoxomil. Chromatographic separation was based on an Agilent Zorbax Eclipse Plus C18(250 mm × 4. 6 mm, 5 μm)column using water(containing 0. 1% formic acid)- acetonitrile as mobile phase in gradient elution mode. Mass spectrometry was operated in positive ion mode. Selective ion monitors were set at m/z 315 for BBT1 and at m/z 395 for BBT2. Good linear correlations were observed in the range of 0. 009 4- 0. 561 0 μg/mL(r=0. 998)with the quantification limit at 9. 35 ng/mL and the detection limit at 3. 12 ng/mL for BBT1, and in the range of 0. 018 2- 0. 547 5 μg/mL(r=0. 999)with the quantification limit at 18. 25 ng/mL and the detection limit at 6. 08 ng/mL for BBT2. Furthermore, the average recoveries of the three spiked concentration level were 96. 5%(n=9, RSD=4. 8%)and 98. 0%(n=9, RSD=5. 1%)for BBT1 and BBT2, respectively. The proposed method is simple, specific and accurate, and quite suitable for the determination of BBT1 and BBT2 in olmesartan medoxomil.

Objective: To explore the utility of circulating tumor DNA detection in early breast cancer by using next-generation sequencing. Methods: This exploratory study of circulating tumor DNA detection is for early invasive breast cancer patients treated in Breast Disease Center, Peking University First Hospital from December 2015 to July 2016. Plasma samples were collected and were used to isolate plasma cell-free DNA.Exons or hotspots of 247 cancer related genes were sequenced by next-generation sequencing. Mutations and their correlation with clinic-pathological factors were analyzed. The correlation between mutations and clinic-pathological factors was evaluated by χ² test or Fisher′s exact test. Results: Seventy-five patients were enrolled in this study. All patients were female and aged from 31 to 88 years with median age of 58 years. All patients′ clinic-pathological records were complete. Sixty-four mutations in 18 genes (ALK, BCR, ERBB2, ROS1, PDGFRA, EGFR, FGFR2, CYP1B1, CALR, CASP7, BRAF, FGFR1, FGFR3, MET, NRAS, PTEN, KIT, SOD2) were detected in 47 (62.7%) among all 75 patients.Exons were captured in 10 genes, and mutations in 2 of 3 genes analyzed were clustered. Gene mutations were not correlated with menopausal status, histological type, primary tumor (T), regional lymph nodes (N), TNM stage, histological grade, estrogen receptor status, progesterone receptor status, human epidermal growth factor receptor 2 status, Ki-67 and molecular subtype (all P>0.05). Conclusion Circulating tumor DNA sequencing by next-generation sequencing was useful for detecting breast cancer-related mutations.

Polycystic ovary syndrome(PCOS)is characterized by high heterogeneity and heredity,and its exact pathogenesis is still not clear.Some studies have shown that epigenetic disorders,such as hyperandrogen-induced methyla-tion or acetylation of lysine at different sites(K4,K9,and K27)in histone H3,methylation and demethylation modifica-tion of genes related to steroids,hormone receptors and follicular development,and transcriptional control of microRNA or long noncoding RNA,play a central role in the occurrence and development of PCOS.This article reviews the research ad-vances in epigenetic mechanisms(histone modifications,DNA methylation,and noncoding RNA)of PCOS,in order to provide a reference for the prediction and early prevention of PCOS.

Objective To identify the surface electromyographic (sEMG) characteristics of bilateral stemocleidomastoid muscles in infants with congenital muscular torticollis (CMT) after rehabilitation.Methods One hundred and twenty two infants were enrolled from Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine between January 2013 and December 2015,including 63 cases receiving rehabilitation (treatment group) and 59 cases without rehabilitation (control group).The sEMG parameters of bilateral sternocleidomastoid muscles in cervical neutral,left and right rotation and stretching posture in the supine position were retrospectively collected before and after treatment/follow-up;and the root mean square (RMS) was analyzed.Results Before treatment,the RMS values of the affected side in cervical neutral position,cervical rotation and stretching posture were significantly lower than those of the unaffected side in both groups(t=15.758 and 11.950,4.871 and 4.746,4.142 and 4.318,P＜0.01).In the follow-up for control group,the RMS values of the affected side in cervical neutral position,cervical rotation and stretching posture were significantly lower than those of the unaffected side (t=10.692,2.198 and 3.239;P＜0.01,=0.03 and ＜0.01).In the treatment group there were no significant differences in the RMS values in cervical neutral position,cervical rotation and stretching posture between affected side and unaffected side after treatment(t=1.647,1.090 and 0.136,P=0.10,0.28 and 0.89).Conclusion Rehabilitation reduces the difference of the RMS values between the affected and unaffected sternocleidomastoid muscles in infants with CMT.

Objective:To investigate the effects of inhibition of Wnt/ β-catenin signaling pathway on paraquat (PQ)-induced transition of human lung fibroblast cell line MRC-5 and related molecular mechanisms.Methods:The MRC-5 cells were divided into three groups. Control group: without drug treatment; PQ group: the cells were treated by PQ (50 μmol/L) for 72 hours to establish cell transition model; PQ+DKK1 group: the cells were treated with PQ (50 μmol/L) and DKK1 (10 ng/mL) for 72 hours. Then, the cells were collected, and the transition related signatures including α-SMA, Vimentin and Collagen I were detected by immunofluorescence and Western blot (WB); Meanwhile, the expression levels of Wnt pathway-related molecules including β-catenin, Cyclin D1 and WISP1 were determined by WB, immunofluorescence and real-time fluorescence quantitative PCR (RT-PCR) during the transition; In addition, the inhibitor of Wnt/β-catenin pathway Dickkopf-1 (DKK1) was applied to block the signaling. Then the expression changes of β-catenin, cyclin D1 and WISP1 were detected by WB to verify the inhibitory effect, and the transition marker molecules including α-SMA, Vimentin and Collagen I were also determined by WB.Results:After 72 hours, compared with the Control group, the expression levels of α-SMA, Vimentin and Collagen I of MRC-5 cells in PQ group were increased significantly ( P<0.05); The expression levels of β-catenin, Cyclin D1 and WISP1 of MRC-5 cells in PQ group were significantly up-regulated ( P<0.05); DKK1 could inhibit the high expression of α-SMA, Vimentin and Collagen I of MRC-5 cells during PQ-induced transition ( P<0.05). Conclusions:DKK1 could inhibit PQ-induced transition of lung fibroblasts by interference with Wnt/β-catenin signaling, which was expected to further inhibit pulmonary fibrosis caused by PQ.

BACKGROUND:The importance of autophagy for maintaining cellular homeostasis and stress response has long been recognized.As a way for cells to selectively clear their damaged organelles to achieve the recycling of cellular components,autophagy has a pivotal role in bone metabolism.OBJECTIVE:To review the role and possible mechanisms of autophagy in regulating bone-related cell activity and function among bone marrow mesenchymal stem cells,osteoblasts,osteocytes,and osteoclasts.METHODS:PubMed was searched for studies related to autophagy using the keywords of "autophagy;bone marrow mesenchymal stem cells;osteoblasts;osteocytes;osteoclasts."RESULTS AND CONCLUSION:We finally included 84 papers.Autophagy plays an important role in bone metabolism.Autophagy is involved in maintaining the balance between mineralization and absorption,and then maintaining bone homeostasis.An appropriate autophagy inducer may also benefit bone remodeling.Abnormal autophagy can lead to disorders of bone balance,leading to diseases such as osteoporosis.We may prevent or treat bone-related diseases by regulating the level of autophagy as its function in maintaining the balance of mineralization and resorption in bone homeostasis.