Objective To establish a stable and useful method for culturing human osteoclast-like cells in vitro,and investigate the effect of 1?,25-(OH)2D3,M-CSF and PGE2 on osteoclasts differentiation,proliferation and activation so as to lay the foundation for further study of the biological mechanism for tooth movement.Methods The HCMNC were isolated and cultured in 24-well plate with coverslips and human dentine slices.The experiment group was cultured with 1?,25-(OH)2D3,M-CSF and PGE2,respectively,while the control group was not.The liquid was changed every 3 days and the whole culture process lasted for 7 days.The phase contrast microscopy and TRAP staining were adopted to identify osteoclast-like cells.Results On the 3rd day the monocytes began to fuse and on the 7th day positive multinucleated cells could be seen with TRAP staining,but absorption pit was not formed on the dentin slices.The group with 1?,25-(OH)2D3 had the largest number of osteoclast-like cells.Conclusion After the monocytes in UCB are cultured by 1?,25-(OH)2D3,M-CSF,PGE2 induction,they can turn into TRAP(+) multinucleate osteoclast-like cells,the 1?,25-(OH)2D3 10-8mol/L being the most effective.

BACKGROUND: Orthodontic tooth movement is dependent on reconstruction of periodontium. Osteoclastic bone resorption is the first step of tooth movement. The present study hotspots focus on signal transduction pathway regarding osteoclast differentiation and functional development under stress and on the relationship between periodontal ligament cells and osteoclasts. OBJECTIVE: To set up a simple method to in vitro culture human osteoclast-like cells and to observe the effects of bone resorption-stimulating factors on differentiation, proliferation, and function of osteoclast-like cells, DESIGN, TIME AND STTING: A cytological in vitro controUod observation was performed at the Central Laboratory,Stomatology Hospital, Xi'an Jiaotong University between October 2007 and May 2008. MATERIALS: Umbilical cord blood was sourced from the healthy puerperae who had not suffered from high-risk pregnancy. Freshly prepared fetal femur provided by Laboratory Animal Center, Xi'an Jiaotong University and were used for preparation of bone flaps at 100-200 μm thickness. 1α ,25-(OH)2D3, macrophage colony-stimulating factor (M-CSF), prostaglandin E2 were purchased from Sigma Company, USA.METHODS: Under aseptic condition, umbilical cord blood was collected. Following Ficoll solution separation and centrifugation, supematant was discarded. Umbilical cord blood-derived mononuclear cells were suspended with o -modified minimal essential medium (α-MEM) solution and then inoculated into a 24-well culture plate, in which, coverslips and femoral slices were pre-placed, at a density of 1×109/L, 1.0 mL per well. Five groups were set, blank control, 108 mol/L 1α ,25-(OH)2D3, 10-7 mol/L 1α ,25-(OH)2D3, macrophage colony stimulating factor (M-CSF), and 1α ,25-(OH)2D3+prostaglandin E2 (PGE2). Each group was cultured for 7 days. MAIN OUTCOME MEASURES: Cellular growth morphology was observed under an inverted microscope; osteoclast-like celt formation was examined by tartrate-resistant acid phosphatase (TRAP) staining; and osteoclastic Howship's lacuna was detected by toluidine blue staining. Any cell with TRAP-positive staining and more than two nuclei was considered osteoclast-like cell and counted. RESULTS: After 3 days of culture, cells from the blank control group did not exhibit apparent changes in morphology and quantity. In the remaining groups, mononuclear cells appeared with confluent tendency. After 7 days of culture, a small number of osteoclast- like cells with 2-3 nuclei were found in the blank control group; a great many of multinucleated osteoclast- like cells with 3-20 nuclei were present in the remaining groups. Through the use of optical microscope, osteoclast-like cells could be found for the presence of red cytoplasm, bright yellow nuclei, and TRAP-positive staining in each inducing factor-treated group, in particular in the 108 mol/L 1α ,25-(OH)2D3 group, which displayed osteoclast- like cells exhibiting 14 nuclei, strong TRAP-positive staining, and a relatively big cell body. But no osteoclastic Howship's lacuna was found in any group. Compared to the blank control group, the numbers of osteoclast-like cells were greater in each inducing factor-treated group (F = 9.78, P < 0.01). There was no significant difference in the numbers of osteoclast-like cells between the 108 mol/L 1α ,25-(OH)2D3 and the 10-7 mol/L 1 a ,25-(OH)2D3 groups (P0.05). The M-CSF group and the 1α ,25-(OH)2D3+PGE2 group exhibited significantly less numbers of osteoclast-like cells than the 108 mol/L 1α ,25-(OH)2D3 group (F= 7.46, P< 0.01). CONCLUSION: After in vitro culture of 1α ,25-(OH)2D3, M-CSF, and PGE2, umbilical cord blood-derived mononuclear cells can differentiate into TRAP-positive multinucleated osteoclast-like cells, the 10-8 mol/L 1α ,25-(OH)2D3 being the most effective.

Objective By culturing the osteoclasts together with the osteoblasts directly to investigate the effect of osteoblasts on the formation of mature osteoclasts.Methods The bone marrow mononuclear cells of rats were treated with 30?g/L M-CSF and 50?g/L RANKL and cultured for 6 days.Subsequently,the primary osteoblasts which were of the same quantity as the osteoclasts were co-cultured directly.In the co-culture system,we added the liquid containing 1,25-(OH)2D3 1?10-8mol/L and PGE2 1?10-6mol/L.The morphological observation,TRAP staining and pit staining were adopted to identify osteoclasts.Results When the osteoclasts were co-cultured with primary osteoblasts,the growth of osteoblasts had more preponderances.After staining,we could see more osteoblasts than osteoclasts.Conclusion The relationship between osteoblasts and osteoclasts is related to the relative quantities of the two cells.When osteoblasts outnumber osteoclasts,osteoblasts would inhibit the formation and differentiation of osteoclasts.

Objective:To study the effects of miR-20a on the osteogenic differentiation potential of inflammatory periodontal liga-ment cells(IPDLSCs).Methods:Cells were isolated and cultured from the healthy and inflammatory periodontal ligament samples (HPDLSCs and IPDLSCs)respectively.miR-20a expression was analyzed by qRT-PCR.Alizarin red staining,Western blot and PCR were used to evaluate the osteogenic differentiation potential of IPDLSCs after transient transinfection of miR-20a mimics or inhib-itor.Results:miR-20a expression in IPDLSCs was lower than that in HPDLSCs,and the osteogenic differentiation potential of IP-DLSCs were promoted by miR-20a mimics,and reduced by miR-20a inhibitor.Conclusion:The miR-20a in IPDLSCs was down reg-ulated.miR-20a can promote the osteogenic differentiation potential of IPDLSCs.

Objective: To observe the effect of electroacupuncture (EA) at different time points during the perioperative period on the recovery of gastrointestinal function after gastrointestinal malignant neoplasms surgery. Methods: Sixty-three patients who needed radical surgery for gastrointestinal tumors were randomized into a control group, treatment group 1 (postoperative EA group), and treatment group 2 (intraoperative and postoperative EA group). The control group received surgery and conventional Western medicine treatment, and treatment groups 1 and 2 received additional EA treatment at different time points. The initial flatus time after the surgery, visual analog scale (VAS) score at different time points after the surgery, the proportion of using patient-controlled analgesia (PCA) after the surgery, and the times of adding analgesics were observed in the three groups. Results: The initial flatus time after the surgery was earlier in treatment groups 1 and 2 than in the control group (P<0.05); the difference between treatment groups 1 and 2 was statistically insignificant (P>0.05). The VAS score was lower in treatment group 2 than in the control group at 6, 12, 24, and 72 h after the surgery (P<0.05); the VAS score was lower in treatment group 1 than in the control group only at 72 h after the surgery (P<0.05). There were no significant differences in the rate of using PCA among the three groups (P>0.05). Regarding the times of adding analgesics, it was less in treatment group 2 than in the control group at 12 h after the surgery (P<0.05). Conclusion: Either EA during and after the surgery or only after the surgery can hasten the initial flatus and boost the recovery of gastrointestinal function in patients after radical resection of gastrointestinal neoplasms. Successive EA during and after the surgery should be superior to postoperative EA regarding the analgesic effect after the surgery.

Objective To compare the expression of histone deacetylase(HDAC)1-11 of humanperiodontal ligament stem cells(PDLSC) in normal and inflammatory microenvironments,and to investigate the effect of histone deacetylase inhibitor trichostatin A(TSA) on the osteogenic differentiation potential of PDLSC in inflammatory microenvironment induced by tumor necrosis factor-α(TNF-α) stimulation.Methods PDLSC were isolated from periodontal ligament tissues obtained from the surgically extracted human teeth and cultured by single-colony selection.The expression of HDAC1-11 in cells with or without TNF-α(10 μg/L) stimulation was evaluated by quantitative real time-PCR(RT-PCR).The effect of TSA on cell proliferation was investigated by methyl thiazolyl tetrazolium(MTT) assay.The influence of TSA on osteogenic differentiation of PDLSC in inflammatory microenvironment with TNF-α stimulation was assessed by alizarin red staining,qoantitative RT-PCR and Western blotting,respectively.Results The expression of HDAC in PDLSC with TNF-α stimulation was significantly higher than that in normal PDLSC(P＜0.05) (except HDAC7,P=0.243).TSA had no significant effect on PDLSC proliferation at the concentration of 50 nmol/L(P=0.232).The alizarin red staining showed that PDLSC in TNF-α group generated less mineralized nodule than the control group,while the cell matrix mineralization in TSA group was improved obviously.TNF-α had an inhibitory effect on the expression of osteogenesis related genes,runt-related transcription factor-2(RUNX2) and alkaline phosphatase(ALP),with relative gene expression ratio (experimental/control) decreased to 0.17 ±0.02 and 0.32±0.03,while TSA could significantly increase the genes' expression to 0.67±0.03 and 0.89±0.02(P＜0.01).Western blotting test showed that in TNF-α group the expression of osteogenesis related proteins was obviously reduced,and compared with the TNF-α group,TSA could significantly promote the expression of proteinsin inflammatory microenvironment.Conclusions PDLSC in inflammatory microenvironment by TNF-α stimulation had a higher expression of HDAC than that in normal conditions.TSA,as a histone deacetylase inhibitor,could significantly promote the osteogenic differentiation potential of PDLSC in inflammatory microenvironment by suppressing HDAC.

Whole blood transfusion for the resuscitation of trauma patients is not a new concept, with its history dating back to World War I. Due to the significant survival benefits of early intervention with whole blood, an increasing number of countries and regions are using whole blood for pre-hospital resuscitation of patients with traumatic haemorrhage. Whole blood containing low-titer anti-A and anti-B antibodies is known as low-titer group O whole blood. The safety of transfusion of low-titer group O whole blood has been proven in military and local trauma centers in some countries. The use of low-titer group O whole blood for pre-hospital trauma care in China will pose new challenges to blood stations that provide whole blood. This paper reviews the selection of group O donors, the setting of anti-A and anti-B titers threshold and their detection, as well as the collection, preparation and storage of whole blood.

Mutations in the mitochondrial DNA have been certified to be one of the most important causes of maternally inherited sensorineural hearing loss. Among these, mitochondrial 12S rRNA1555A>G, 1494C>T and other mutations are associated with both nonsyndromic and drug induced hearing loss caused by aminoglycosides. Individuals carrying 1555A>G or 1494C>T mutation have a variety of clinical manifestations, which implies that the 1555A>G or 1494C>T mutation is a chief factor underlying the development of deafness but insufficient to produce the clinical phenotype. Therefore other modifier factors, such as aminoglycosides, mitochondrial haplotypes, secondary mutation or nuclear modifier genes, may play an important role in the phenotypic expression of the deafness-associated mitochondrial 12S rRNA1555A>G or 1494C>T mutation. In this review, the modifier factors for the phenotypic expression of deafness-associated mitochondrial 12S rRNA1555A>G or 1494C>T mutations were summarized and proposed the pathogenesis of maternally inherited deafness.

Animals ; Antibodies, Neutralizing/biosynthesis* ; Antibodies, Viral/biosynthesis* ; Body Weight/immunology* ; COVID-19/virology* ; Disease Models, Animal ; Disease Progression ; Humans ; Immunohistochemistry ; Lung/virology* ; Male ; Mesocricetus ; Nasal Cavity/virology* ; RNA, Viral/immunology* ; SARS-CoV-2/pathogenicity* ; Severity of Illness Index ; Viral Load

ObjectiveTo investigate an outbreak of norovirus diarrhea in a welfare home for the elderly in Shanghai， and to analyze the epidemiology and virus genotype characteristics of the epidemic， thus providing a scientific basis for the prevention and control of norovirus epidemic. MethodsCase definition was clarified. After case definition， a standardized questionnaire was used to carry out case investigation to make comprehensive analysis in combination with field epidemiological investigation and laboratory tests results. ResultsThe epidemic lasted for 14 days， and 23 cases were reported with an incidence rate of 12.37% （23/186）， including 3 males and 20 females. There were more cases on the 1st， 4th and 6th floor of the main building in the welfare home， accounting for 52.17% （12/23） of total cases. 19 cases were reported in the main building， with an incidence rate of 11.59% （19/164）； 4 cases were reported in the auxiliary building， with an incidence rate of 18.18% （4/22）. There was no significant difference in the morbidity between the main building and the auxiliary building （χ²=0.779，P>0.05）. The main clinical manifestations were vomiting and diarrhea. There was a significant difference in the incidence of vomiting symptoms among the elderly， nursing staff and other staff in the welfare home （χ²=10.216， P<0.05）. But there was no significant difference in the incidence of diarrhea among the elderly， nursing staff and other staff （χ²=1.218， P>0.05）. Fecal samples were collected from 23 cases， 1 case family member， 68 risk personnel and 14 environmental surface swab samples. Norovirus GⅡ was detected in stool samples of 19 cases， 1 family member and 36 risk personnel. ConclusionOutbreak of norovirus infection is reported in a welfare home in Shanghai. The close contact between the elderly and health workers might lead to the outbreak.