Objective To establish osteoblasts osteoclasts co culture system in vitro for the study of the biological characteristics of osteoblasts and osteoclasts and also to analyze the interactions between osteoblasts and osteoclasts. Methods The osteoblasts and osteoclasts were isolated from cancellous bone in illium. Osteoblasts osteoclasts co culture system was built to prevent these two kinds of cells from contact with each other although with the facility of their medium exchange. The osteogenety of osteoblasts was evaluated by MTT and ALP activity; the resorption activity of osteoclasts was investigated by TRAP activity and the area of absorption lacuna. Results Osteoblasts are full fusiform, ALP stain positive cells, while osteoclasts are multinuclear and TRAP stain positive, with the ability of forming bone lacuna; while in the co culture system, the proliferation of osteoblasts in co culture (0.60?0.08)OD is quicker(P=0.000)than that in mono culture (0.36?0.03)OD, and the ALP activity increased significantly (P=0.000)from (18.33?0.34)u/mg to (23.37?2.48) u/mg; while the average area of bone resorption formed by osteoclasts [(6.55?0.34)?10-2] ?m2 was also increased obviously(P=0.000)from [(5.15?0.17)?10-2] ?m2. Conclusion In co culture system, osteoclasts were able to enhance ossification effects of osteoblasts by accelerating their proliferation and differentiation; at the same time, osteoblasts upregulated the ability of bone resorption of osteoclasts. This implied that osteoblasts and osteoclasts can promote each others function. This study provide reliable model for bone metabolism research.

The ketoreductase (KR) domain in the first extending module of the polyketide synthase (PKS) catalyzes the reductions of both an α-keto group and a β-keto group in the biosynthesis of bacillaene, suggesting the intrinsic substrate promiscuity. In order to further investigate the substrate specificity, the KR domain (BacKR1) was heterologously overexpressed in Escherichia coli. In vitro enzymatic analysis showed that only one of the four diastereomers was formed in the reduction of the racemic (±)-2-methyl-3-oxopentanoyl-N-acetylcysteamine thioester catalyzed by BacKR1. In addition, BacKR1 was revealed to catalyze the reductions of cyclohexanone and p-chloroacetophenone, indicating the potential of KR domians of PKSs as biocatalysts.
Bacterial Proteins ; genetics ; metabolism ; Catalysis ; Cyclohexanones ; metabolism ; Escherichia coli ; enzymology ; Polyketide Synthases ; genetics ; metabolism ; Protein Structure, Tertiary ; Substrate Specificity ; omega-Chloroacetophenone ; metabolism

Objective To evaluate the values of combined detection of soluble mesothelin related proteins (SMRP)and carbohydrate antigen 1 25 (CA1 25)in the diagnosis of malignant peritoneal mesothelioma.Methods 20 patients with malignant peritoneal mesothelioma were selected as the malignant group,50 patients with benign ovar-ian tumor were selected as the benign group,60 healthy women were selected as the control group.Enzyme -linked immunosorbent assay(ELISA)was adopted to detect serum SMRP levels and electrochemiluminescence immunifaction (ECLI)was adopted to detect serum CA1 25 levels.The SMRP,CA1 25 levels were compared among all groups,the sensitivity,specificity of SMRP and CA1 25 in the diagnosis of peritoneal malignant mesothelioma were analyzed. Results Serum levels of SMRP,CA1 25 in malignant group were (1 4.8 ±1 .6 )μg/L,(1 89.1 ±1 5.3 )μg/Lrespectively,which were significantly higher than those in the benign group[(3.9 ±0.9)μg/L,(28.6 ±4.2)μg/L] (t =27.40,49.61 0,all P <0.01 )and the control group[(2.8 ±0.7)μg/L,(1 6.9 ±3.8)μg/L](t =29.877, 53.334,all P <0.01 ).The sensitivities of SMRP,CA1 25 detection alone for malignant peritoneal mesothelioma diag-nosis were 65.0%,50%,respectively,while their specificities were 83.6%,79.1 %.The sensitivity and specificity of combined detection of SMRP and CA1 25 for malignant peritoneal mesothelioma were 95.0% and 93.6%.Conclusion SMRP has some value in the diagnosis of peritoneal malignant mesothelioma.SMRP combined with CA1 25 detection can markedly improve the diagnostic sensitivity and specificity for malignant peritoneal mesothelioma.

Objective To compare the clinical effect of different doses of methylprednisolone in the treatment of children with severe hand foot mouth disease research and analysis. Methods 100 patients in our hospital from January 2014 to August 2016 were selected and randomly divided into the control group and the experimental group, with 50 patients in each group. The control group were treated with small doses of methylprednisolone, 2 mg/ (kg?d), intravenous injection. The experimental group was treated with large dose of methylprednisolone, 10～15 mg/(kg?d) intravenous infusion. The therapeutic effects of the experimental group and the control group were compared and analyzed. Results after the corresponding treatment, the number of adverse reactions in the experimental group was 31, the adverse reaction rate was 62.0％, and the adverse reaction rate in the control group was 60.0％. There was no significant difference in the incidence of adverse reactions between the two groups. The upper respiratory rate and critical illness in the experimental group were 26.0％ and 32.0％, significantly lower than that of the control group (P<0.05). The average remission time of the patients in the experimental group was (2.19±1.01) days. The symptomatic remission time in the control group was (4.19±1.89) days. The remission time of the control group was significantly longer than that of the experimental group, with statistical difference (P<0.05). Conclusion The short term large dose of methylprednisolone in treatment of children with severe HFMD clinical effect is ideal, can be reduced to critical illness rate, less symptom remission time, with the further promotion of the clinical significance.

Objective To establish a sensitive,valid and rapid high performance liquid chromatography-trap mass spectrometry (HPLC-MS) method for determining carbocisteine concentration in human plasma. Methods Plasma samples were pretreated by 100 mL/L perchloric acid to remove the interference protein. The supernatant was separated by reverse phase HPLC,electrospray MS was used to detect carbocisteine in plasma samples,and selective ion method of multiple reaction modes was used to determine the concentration of carbocisteine. Results The limit of detection was 0.01 ?g/mL and the linear range was 0.2-200.0 ?g/mL. The recovery rate was within the range of 89.2%-105.6%. Conclusion The proposed method could be used to determine carbocisteine in biological samples.

[ ABSTRACT] AIM:To cultivate stem-like spheres from SW620 cell line in the specific serum-free medium and evaluate the features of the cancer stem cells, and to investigate the effects of docosahexaenoic acid ( DHA) and eicosapen-taenoic acid ( EPA) on the growth of SW620 stem cell-like cells.METHODS: Human colon cancer stem cell-like cells ( CSCLC) were obtained from SW620 spheres cultured in serum-free medium.These cells were tested for the expression of SSEA-1 and TRA-1-81 by immunofluorescence staining.The mRNA expression of Sox-2 and Oct-4 was detected by real-time PCR.The efficiency of colony formation on a soft agar gel and tumor formation in the nude mice was compared between SW620 adherent cells and CSCLC.The inhibitory effects of 5-fluorouracil (5-FU) and mitomycin C on both types of cells were measured by MTS assay.MTS assay, Annexin V/PI staining and trypan blue staining were used to determine the effects of DHA and EPA on both types of cells.MTS assay was also used to analyze the combined effect of DHA or EPA with chemotherapeutic drugs on SW620 CSCLC.RESULTS:SW620 cells formed spheres in serum-free culture.The cells from spheres highly expressed SSEA-1 and TRA-1-81, transiently expressed Sox-2 and Oct-4 genes and were more resistant to 5-FU and mitomycin C treatments.These cells exhibited a greater ability in clone formation and tumorigenicity, indica-ting that these cells carried stem cell-like features, hence were considered SW620-derived CSCLC.DHA and/or EPA sup-pressed SW620 CSCLC by inhibiting cell growth, inducing cell apoptosis and sensitizing them to chemotherapeutic drugs. CONCLUSION:The cells with stem cell-like features, such as high efficiency in clonogenicity, tumorigenicity and resist-ance to chemotherapeutic drugs, can be obtained from SW620 spheres cultured in serum-free condition.DHA and EPA in-duce apoptosis in SW620-derived CSCLC and sensitize them to chemotherapeutic drugs.

Objective To compare the errors of double-center and single-center setup, and to study the role of both on reducing the rotational setup errors for the patients with esophageal carcinoma depend on rigid registration errors between online kV-cone-beam computed tomography (kV-CBCT) images and plans for CT images. Methods 20 patients with middle esophageal carcinoma received image scanning before treatment every week by using double-center setup and CBCT, and single-center setup images of 20 patients were taken from the X volume image (XVI) system. Then the images of both setup types, registration errors of CT image and rotational setup errors were compared respectively. Every patient received kV-CBCT scanning analysis before treatment every week, and 6 times in total. 240 group of kV-CBCT images from all of the patients were off-line matched with plans for CT images to calculate the errors of X-axis, Y-axis, Z-axis. Then the data of linear errors and rotational setup errors from patients were collected, aiming at putting the error data into the patients treatment program and analyzing the significances. Results The standard registration of double-center setup was as follows: T (X) (0.28 ±0.19) cm, T (Y) (0.27 ±0.19) cm, T (Z) (0.33 ±0.12) cm, R (X) (0.40 ±0.19)° , R (Y) (0.30 ±0.18)° , R (Z) (0.30 ±0.19)° . The standard registration of single-center setup was as follows:T(X) (0.32±0.20) cm, T(Y) (0.29±0.25)cm, T(Z) (0.31±0.16) cm, R(X) (2.2±0.68)°, R(Y) (0.5±0.32)°, R(Z) (2.10±0.60)°. There were statistical differences between linear errors in T(X) and rotational setup errors in R(X), R(Y) or R(Z) (P< 0.05). Conclusion Double-center position can reduce the rotational setup errors, especially in X-axis, Y-axis errors, and may provide more help for the radiation oncology departments without on-board CBCT.

The levels of total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, apolipoprotein (APO) A1 and APO B were lower in Bai Ku Yao than those in Han nationalities (all P<0.01). There was no significant difference in serum triglyceride levels and the ratio of Apo A1 to Apo B between two nationalities. Dyslipidemia was positively correlated with body mass index, waist circumference, total energy and total fat intakes, and inversely correlated with degree of physical activity and total dietary fiber intake in both ethnic groups. In addition, dyslipidemia was also positively correlated with age and alcohol consumption in Han, but not in Bai Ku Yao.

Pinch-3 protein is an important constituent of cell membranes, which directly affects the cell morphology and mechanical properties. We observed and compared the change of morphology and cell traction force of glomerular podocytes before and after Pinch-3 gene inhibition by gene interference technology in this experiment. We found that a number of pores appeared on the cell surface, and the cell projected area were increased at the same time, with an approximate average about an increase of 40% after Pinch-3 gene inhibition. The results showed that the cell traction force of glomerular podocytes was significantly reduced, with an approximate average decrease of 40%, the maximum value of the cell traction force was reduced and the distribution of cell traction force became dispersive. All this suggested that after Pinch-3 gene inhibition, some pores created on the cell surface influenced the physical properties of glomerular podocytes and then affected the cell projected area and influenced the formation and distribution of cell traction force of the glomerular podocytes as well.
Adaptor Proteins, Signal Transducing ; genetics ; physiology ; Biomechanical Phenomena ; Cell Movement ; Genetic Engineering ; Humans ; Kidney Glomerulus ; cytology ; LIM Domain Proteins ; genetics ; physiology ; Mechanotransduction, Cellular ; physiology ; Membrane Proteins ; genetics ; physiology ; Podocytes ; cytology ; physiology ; Stress, Mechanical

Objective To compare the size of target volume,amplitudes of movements in different directions,movement vector,dose to the diseased lung,whole lung volume,and setup error between free breathing fixation (method A) and four-dimensional computed tomography (4DCT)-guided abdominal balloon compression fixation (method B),and to demonstrate that the 4DCT-guided abdominal balloon compression fixation is effective in the treatment of non-small cell lung cancer (NSCLC).Methods A retrospective analysis was performed among 80 patients with NSCLC in our hospital.In those patients,40 received method A and 40 method B.The GTVfree and GTVpress were delineated on the maximum intensity projection (MIP) images of 10 respiratory phases using method A and method B,respectively.The PTVfree and PTVPress were obtained by expansion of the GTVfree and GTVpress,respectively.The paired t test was used to analyze the differences in the PTV,maximum amplitudes of movements in three dimensions,absolute value of the movement vector (|V|),and volume between method A and method B.The treatment planning system was used to compare the V5,V10,V20,and V30 of the diseased lung and the whole lung volume between method A and method B.All patients underwent cone-beam CT (CBCT) scans after positioning.Setup error was obtained by matching the CBCT images with the MIP images in the XVI system based on bone and grayscale values.Results The PTVfree and PTVpress were (283.2± 12.74) and (201.8± 12.99)cm3,respectively (P=0.002).The maximum amplitudes of movements in the right-left,superior-inferior,and anterior-posterior directions as well as thel V | value were (0.22±0.02),(1.85±0.08),(0.43±0.26),and (1.91±0.27) em,respectively,for method A,and (0.05±0.01),(0.41±0.03),(0.16±0.16),and (0.44±0.16) cm,respectively,for method B (P=0.120,0.001,0.070).The V5,V10,V20,and V30 for the diseased lung and total lung volume were (61.26± 4.27) ％,(44.52± 1.70) ％,(28.22± 3.13) ％,(18.26±5.17)％,and (3556±223.12) cm3,respectively,for method A,and (52.74±4.78)％,(38.76±4.92) ％,(23.71 ±4.03) ％,(15.54±3.43) ％,and (3376±311.65) cm3,respectively,for method B (P =0.001,0.003,0.004,0.021,0.004).There was no significant difference in setup error obtained by the XVI system between the two fixation methods (P＞0.05).Conclusions Without increasing setup error,abdominal balloon compression can effectively control the lung movement amplitude,reduce the planning target volume,and reduce the radiation dose to the lung in patients with NSCLC.