BACKGROUND: Studies have found that miR-223-3p is highly expressed in osteoporosis patients, but the related mechanism is rarely studied. OBJECTIVE: To investigate the regulatory effect of hsa-miR-223-3p on osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells were isolated and obtained by Percoll density gradient centrifugation. The cells were induced into osteogenic differentiation, and overexpressed hsa-miR-223-3p. RT-qPCR and western blot assay were used to detect the changes in the mRNA and protein levels of Wnt signaling pathway markers Wnt5a and its downstream signaling molecules OPG and RUNX2. PmirGLO/Wnt5a-3'UTR and pmirGLO/Wnt5a-3'UTR mut plasmids were constructed and co-transfected with hsa-miR-223-3p mimics/control plasmid into 293T cells to verify the targeting relationship between hsa-miR-223-3p and Wnt5a by dual luciferase reporter gene system. RESULTS AND CONCLUSION: (1) With the prolongation of osteogenic induction time, the level of alkaline phosphatase in cell culture fluid gradually increased. The deposition of dark red or reddish brown calcium salt nodules was gradually increased according to alizarin red staining results. (2) Overexpression of hsa-miR-223-3p decreased the mRNA and protein levels of Wnt5a, OPG and RUNX2 in cells. (3) hsa-miR-223-3p directly targeted the 3'UTR region of Wnt5a and reduced the luciferase activity of Wnt5a. This targeting effect disappeared when the binding site of Wnt5a was mutated. Blocking hsa-miR-223-3p increased the luciferase activity of wild-type Wnt5a, while the luciferase activity of mutant Wnt5a remained unchanged. (4) The results showed that during the differentiation of bone marrow mesenchymal stem cells into osteoblasts, overexpression of hsa-miR-223-3p could affect the expression of Wnt signaling pathway marker Wnt5a and its downstream signaling molecules OPG and RUNX2. hsa-miR-223-3p could directly bind to Wnt5a, which was negatively regulated as a downstream target gene of hsa-miR-223-3p.

Objective To explore the effect of follow-up nursing intervention on preventing dental plaque during oral fixed orthodontic appliance.Methods One hundred and twenty orthodontic patients receiving oral fixed orthodontic treatment were randomly divided into control and experiment groups equally according to visit time.The control group received conventional nursing during the process and follow-up nursing intervention during the process.The two groups were compared in terms of dental plaques before orthodontic, 3 and 6 months after the orthodontic treatment.Results Before the treatment, there was no significant differences in the dental plaques between them(P>0.05), but the indices of dental plagues in the experimental group were significantly lower than those of the control group 3 and 6 months after the orthodontic appliance(P<0.001).Conclusion Follow-up nursing care during orthodontic appliance are more effective for keeping oral hygiene.

Objective To explore the effect of different type of atrial fibrillation(AF) on plasma brain natriuretic peptide(BNP) level in male patients.Methods Fifty-eight hospitalized male patients with AF were enrolled,including 35 cases with paroxysmal AF,14 cases with persistent AF,and 9 cases with permanent AF.All subjects had the examination of echocardiography/electrocardiogram within 24 h after admission.Venous blood sample were collected in a quiet prostration state on the next early morning for BNP detection.The plasma BNP level was determined by radioimmunology-assay(RIA).Results The average level of plasma BNP in male patients with permanent AF(2343.25 ± 820.24)μg/L was significantly higher than those with persistent AF([576.43 ± 223.07]μg/L,t=8.74,P＜0.001) and paroxysmal AF([632.74 ± 103.93]μg/L,t=8.73,P＜0.001),but there was no significant difference between the last two groups(t=0.59,P＞0.05).Conclusion BNP is a sensitive indicator of early heart function impairment related with AF,particularly elevates in male patients with permanent AF.

Objective:To study on effect of Danshao Huaxian Capsule on proliferation of activated hepatic stellate cells(HSCs)in vivo and in vitro.Methods:(1)In vitro experiment:57 male Wistar rats were divided into a normal group,a model group and a treatment group.The rats of model group and treatment group were made hepatic fibrosis model with complex stimulation of CCl_4,drinking alcohol,high lipids and low protein diet for 8 weeks.The treatment group were treated for 8 weeks with intragastric perfusion of Danshao Huaxian Capsule,1g/kg.At the end of experiment,a part of the rats were used for detection of hepatic functions and hepatic fibrosis degrees,and another part of the rats were used to separate HSCs.Cell cycle percentage was detected with flow cytometry.(2)In vivo experiment:prepare Danshao Huaxian Capsule serum of the rat;separate cultured HSCs of the rat and divide into calf serum group,normal rat serum group and Danshao Huaxian Capsule serum group.5%,10% and 20% of the above serum was respectively added into the cultured HSCs.Primary HSCs proliferation was detected with MTT method.Results:(1)In vitro experiment:The hepatic function,hepatic fibrosis level and HSCs proliferation reduced significantly.(2)In vivo experiment: Danshao Huaxian Capstde serum group could significantly inhibit proliferation of HSCs as compared with calf serum group and normal rat serum group at the same content.The action showed dose-dependence.Conclusion:Danshao Huaxian Capsule can obviously inhibit proliferation of HSCs in vivo and in vitro.

Objective To summarize the causes and the prevention of bile duct injuries during laparoscopic cholecystectomy (LC). Methods Clinical records of 1 000 cases of LC were analyzed retrospectively. Results Conversions to open cholecystectomy were required in 15 cases(1.5%). Complications took place in 6 cases(0.6%), involving 3 cases of common bile duct injury, 1 case of stomach puncture, 1 case of port hemorrhage, and 1 case of bile leakage. No long-term complications were seen. Conclusions The most frequent cause leading to bile duct injuries is pulling the bile duct to be angulated.

Objective To investigate the effects of c-myc promoter binding protein(MBP-1)gene expression silencing on the pro-liferation in vitro in human gastric cancer cell line SGC-7901.Methods The cells divided into three groups:blank control group (cells without transfecting gastric cancer cell),negative control group(cells transfecting missense sequence)and experimental group (cells transfecting MBP-1 shRNA).Two MBP-1 shRNA sequences and one negative control shRNA sequence were designed,syn-thesized and cloned into pSIREN-retroQ plasma.Then the recombinant plasmids were constructed and transfected into human gas-tric cancer SGC-7901 cells by Lipofectamine 2000.After selection of cells with puromycin for two weeks,stable cell line SGC-7901 expressing gene of MBP-1 specific shRNA was obtained.The expressions of MBP-1 mRNA and protein in SGC-7901 were deter-mined by the real time PCR and Western blot,respectively.The effects of altered expression of MBP-1 on the cell proliferation were measured by MTT cell proliferation assay.Results PCR and sequencing indicated that the recombinant plasmids pSIREN-retroQ was constructed.Then the recombinant plasmids were transfected into human gastric cancer SGC-7901 cells by Lipofectamine 2000. After selection of cells with puromycin for two weeks,stable cell line SGC-7901 expressing gene of MBP-1 specific shRNA was ob-tained.The relative expression level MBP-1 mRNA in the MBP-1 siRNA transfection group was significantly decreased compared with the blank control group(P <0.05).Compared with the blank group,the expression levels of MBP-1 protein in the experimental group also significantly decreased.The proliferation abilities of SGC-7901 cells at 48,72,96,120 h after MBP-1 siRNA transfection were significantly increased compared with the blank control group (P < 0.05 ).Conclusion Down-regulating the expression of MBP-1 can obviously promote the proliferation of human gastric cancer cell line SGC-7901.MBP-1 gene may become the new target of gene therapy for gastric cancer.

Accidents, Occupational ; prevention & control ; Occupational Diseases ; prevention & control ; Occupational Health Services ; organization & administration ; Risk Assessment ; methods ; organization & administration

Objective To analyse the clinical data of 77 patients with unsuspected gallbladder carcinoma,and to determine the impact of TNM stage and the timing of the second operation on postoperative survival.Methods A retrospectively analysis of 77 patients operated between January 2008 and January 2013 in our hospital for unsuspected gallbladder carcinoma was carried out.The case inclusion criteria was in strict accordance with the medical records which described without preoperative diagnosis,or cholecystectomy for benign gallbladder diseases,or intraoperative or postoperative pathological diagnosis of gallbladder cancer.References to previous published medical literature and the intervals from initial cholecystectomy to further treatment were analyzed to see whether the prognosis and survival varied depending on the TNM staging and whether secondary radical surgery was carried out.Results For the 77 patients in this study,they all had high risk factors associated with gallbladder cancer which included:women,aged ＞ 50 and gallbladder stones.The initial surgery included laparoscopic cholecystectomy (n =53),and open cholecystectomy (n =24).In 26 patients,intraoperative frozen section confirmed the diagnosis and they were treated according to the TNM staging (radical operation n =7,palliative surgery n =17).The postoperative 1-,2-,3-year cumulative survival rates were 65％,45％ and 20％,respectively.For the 54 patients who underwent radical or extended radical cholecystectomy within a short interval from the first operation,the 1-,2-,3-year cumulative survival rates were 82.5％,62.5％ and 45.7％,respectively.Conclusions For resectable gallbladder cancer,the prognosis of unsuspected gallbladder carcinoma was related not only to the clinical staging,but also to the timing of the radical operation.The shorter time interval,the longer the survival,and the better the prognosis.There was no significant difference in prognosis for the group of patients with time intervals between the two operations of less than two weeks when compared with the group with immediate radical surgery.For advanced stages of gallbladder cancer,palliative surgery should be given according to the patient's general condition,aiming to improve quality of life.

Objective To verify the accuracy of blood oxygenation level dependent (BOLD)-based activation using electrocortical stimulation mapping (ESM) and explore the value of language fMRI in the navigating operation of neurosurgery. Methods In 8 cases with brain tumors,BOLD-fMRI examinations were done before the operations. Under the state of awake anesthesia,the patients were aroused and ESM was conducted. Point-to-point comparison between the BOLD signal activations and the ESM was carried out under the surveillance of the neuro-navigation technology. In order to observe the sensibility and specificity of BOLD activations, the location of BOLD activations and the point of ESM was compared to calculate the stimulating positive points inside the regions of BOLD signals(real positive), outside BOLD regions(pseudo-negative), the stimulating negative points inside the regions of BOLD signals(pseudo-positive), and outside BOLD region(real negative). Two kinds of criteria for assessment were used. One was that the positive stimulating points were located in BOLD regions, and the other was that the positive stimulating points were located within 1 cm around the range of BOLD regions. Removal of the lesions were conducted with the tissue 1 cm around the language region preserved, and the cortex inside 0.5-1.0 cm distance from the positive points were retained. Results Of the 8 cases, only 6 finished the tasks. Among them, 3 cases were with astrocytoma of grade 2,2 were with astrocytoma of grade 3, and one with glioblastoma. The total number of stimulating points was 48, among which the positive points were 11. When the first criteria was applied, the sensitivity was 72.7% (8/11), and the specificity was 81.8% (30/37). When the second criteria was applied, the sensitivity was 82.0% (9/11),and the specificity was 75.6% (28/37). Follow-up after operation showed no aphasia occurred. Conclusions BOLD-fMRI had a high sensitivity and specificity in displaying the language regions. But due to the great variation of brain language area among the people, we need more studies of large sample to obtain enough experience before it can be used clinically.

Objective (1) To determine the relationship between the MR signal intensity and the actual flow velocity under steady flow condition (2) to analyze the influence produced by the angle (?) between the flow direction and the velocity encoded gradient direction, and flip angle as well as section thickness on the velocity measurement under oblique flow condition (3) to develop a suitable protocol for using this method to measure volumetric flow rate in the transverse sinus system Methods Flow phantom, which simulated blood flow conditions in the transverse sinus system, consisted of a fluid filled cylinder and a bent tube with a 3 4 mm internal diameter A 1 5 T superconductive MR imager and VIGRE sequence were used for these studies A suitable protocol was based on consideration of the effects of (1) the accuracy of velocity and transverse area measurement of flow, and (2) signal to noise ratio (SNR) Results (1) Signal intensity (y) determined by MR and the actual flow velocity (x) showed straight line correlation, y=68 914x+357 206, R 2=0 998 (2) As the angle (?) increased, the transverse area of the signal determined by MR also increased, but the value of flow velocity decreased (3) As the flip angle increased, the SNR varied from 5 7 to 11 2 The maximum SNR was obtained with 30?flip angle (4) As the section thickness increased, the SNR and the transverse area of the signal determined by MR slightly increased Conclusions Phase contrast MR imaging is a practical method for measuring volumetric flow rates The angle (?) influenced the accuracy of flow velocity and the measurement of transverse area of flow whereas the flip angle and the section thickness substantially influenced the signal to noise ratio and the transverse area of flow