Objective: To study the osteoinductive activity of chimeric molecule of bone morphogenetic protein(BMP)2 and BMP7 expressed in mammalian cells. Methods: Sequences encoding mature peptides of BMP2 and BMP7 were separately amplified by PCR and then linked by overlap-extension PCR with a DNA sequence encoding a flexible peptide (Gly_4Ser)_3 between them. The chimeric DNA sequence was cloned into secretory expression plasmid pcDNA3/sec and then the recombinant plasmid pcDNA3 -BMP2/7 was transfected into CHO-K1 cells. In the presence of G418,cells that stably expressed BMP2/7 were screened out. Thereafter, the conditioned culture medium of the transfected cells was collected and used to treat C3H10T1/2 cells. RT-PCR was employed to study the activity of the recombinant product in inducing osteoblast differentiation. Results: The expression products of chimeric BMP2/7 significantly enhanced the mRNA expression levels of osteoblast phenotype genes, such as alkaline phosphatase, osteocalcin and osteoblast specific transcription factor runt-related transcription factor 2 in C3H10T1/2 cells(P < 0.01). Conclusion: The chimeric expression products of BMP2/7 are capable of forming heterodimers and thus effectively induce non-bone derived cells to differentiate into osteoblasts.

Objective:To investigate the changes of default network functional connectivity (FC) in resting state of patients with insomnia disorder (ID).Methods:Fifty-six patients with insomnia disorder and fifty healthy controls were recruited. All subjects were assessed with Pittsburgh sleep quality index (PSQI), insomnia severity index (ISI), Epworth sleepiness score (ESS) and Hamilton depression scale (HAMD-17). All subjects were scanned with resting state functional magnetic resonance imaging (rs-fMRI). SPM12 and CONN18b were used to preprocess rs-fMRI data on MATLAB (R2013b).The medial superior frontal gyrus was defined as the seed point, and the differences in the functional connection strength of the two groups of subjects were compared. Pearson correlation analysis was performed between the FC value of the brain area with statistical significance and scores of PSQI and ISI.Results:(1)FC analysis showed that compared with the control group, patients with ID had abnormal default mode network (DMN) connection, as follows: FC of left medial superior frontal gyrus and left central anterior gyrus (MNI: x, y, z=-30, -15, 51) and anterior cingulate gyrus (MNI: x, y, z=-6, 24, 36), FC of right medial forehead between the last gyrus and the left central anterior gyrus (MNI: x, y, z=-48, -6, 48), FC of left anterior cuneiform and the left central back (MNI: x, y, z=-54, -18, 54) and left superior occipital gyrus (MNI: x, y, z=-21, -69, 30), FC of right precuneus and left central posterior gyrus (MNI: x, y, z=-60, -21, 48) was enhanced. The FC of left anterior cuneiform lobe and the right middle frontal gyrus (MNI: x, y, z=42, 33, 42) and the right angular gyrus (MNI: x, y, z=54, -51, 45), FC of right precuneus and left inferior temporal gyrus (MNI: x, y, z=-51, -12, -42) was weakened. (2)With the left medial superior frontal gyrus as the seed point, FC values in anterior cingulate gyrus ( r=0.451, 0.338, both P<0.01) and left anterior central gyrus ( r=0.324, 0.402, both P<0.05) were positively correlated with PSQI and ISI scores. With the right precuneus as the seed point, FC value of left posterior central gyrus was positively correlated with PSQI( r=0.333, P=0.013) and ISI scores( r=0.418, P=0.008), while FC value of left inferior temporal gyrus was negatively correlated with PSQI( r=-0.662, P=0.001) and ISI scores( r=-0.402, P<0.01).With the left precuneus as the seed point, FC value of left superior occipital gyrus was positively correlated with PSQI( r=0.438, P=0.001) and ISI scores( r=0.495, P=0.011). Conclusion:Patients with ID may have changes in the functional connection of multiple networks. The decrease of FC in DMN may be one of the potential causes of insomnia. The increase in FC between DMN and the visuospatial attention network may be the core of the mechanism of damage to the brain function network of insomnia link.

Object To develop a new method of discriminating the principle components of Radix Paeoniae Rubra (RPR) from different areas, and to find the reason for forming high-quality of RPR in the place of the genuine. Methods Using Foruier transform infrared (FTIR) absorption spectrometry, the characteristic peaks of fingerprint infrared spectra of RPR samples from 18 habitats were recognized and compared. Results Frequency, intensity and shape of infrared absorption spectra were obviously different between wild and cultivated groups of RPR. The infrared absorption peaks of RPR in Duolun, a famous Chinese orthodox drug, were of distinct characteristics. Conclusion FTIR technique is first applied to rapid analysis of principle component of RPR from different areas. So an operable method in the quality control and discrimination of RPR in the place of the genuine is provided.

Objective To summarize the experiences of extracorporeal membrane oxygenation (ECMO) technique in human lung transplantation. MethodsECMO was used in 45 patients undergoing lung transplantation.There were 32 cases of dilated pulmonary fibrosis and pulmonary hypertension,4 cases of chronic obstructive pulmonary disease,2 cases of pneumosilicosis,2 cases of phthisis,one case of diffuse panbronchiolitis,2 cases of bronchiectasis and 2 cases of primary pulmonary hypertension. In 3 patients, ECMO conduit was used as a bridge to the lung transplantation for 13,19 and 6 days respectively before operation,and ECMO support was done after the operation.In 40 patients ECMO support was done during operation.In 2 patients without using cardiopulmonary bypass, ECMO support was done after operation because of primary graft dysfunction.Results In 40 patients undergoing ECMO support during operation,the ECMO conduit was removed in 37 patients immediately after the operation.ECMO mean blood flow was (2.8 ± 0.6)L/min,running time was (7.7 ± 0.9) h,and 31 patients survived. In 3 patients undergoing postoperative ECMO support,ECMO was removed in 2 cases at 36th h and 6th day after the operation respectively,and the remaining one died from acute renal failure 2 weeks after the operation though the continuing hemofiltration was applied through ECMO conduit. In 2 patients without using cardiopulmonary bypass and 3 patients using ECMO during operation,ECMO support was used initially or again after operation because of PGD or acute heart failure.Three patients were cured,and 2 patients were died from multiple system organ failure.Conclusion ECMO is an important auxiliary tool in lung transplantation for preoperative,intraoperative and postoperative supports. As the transition bridge to recipients,ECMO can be used for preoperative supports; during operation,it can stabilize the haemodynamics,and postoperation,it can improve severe pulmonary function dysfunction and PGD.

Objective To summarize the perioperative clinical effect of extracorporeal membrane oxygenation (ECMO) technique in bilateral lung transplantation of patients with idiopathic pulmonary arterial hypertension.Methods Of 18 recipients with idiopathic pulmonary arterial hypertension receiving double lung transplantation,there were 11 cases of 11 grade Ⅲ (WHO cardiac function ratings) and 7 cases of grade Ⅳ.All patients were given ECMO technique after anesthetic induction,and the speed and volume of ECMO were adjusted according to oxygenation and cardiac function.At the end of the operation,the ECMO flow rate was decreased to 1 L·min-1 ·m-2.If the hemodynamics was stable and the oxygenation index was above 300,the ECMO could be removed.All data were Collected at 7 time points (preoperation,30 min after two lung ventilation,15 min after ECMO,the establishment of one lung ventilation,30 min after pulmonary artery occlusion,30 min after pulmonary artery open and at the end of the operation).The prognosis of the recipients was observed postoperatively.Results All patients successfully completed operation under the support of ECMO technique.As compared with preoperation,PaO2 and PaCO2 were significantly increased at 30 min after two lung ventilation (P＜0.05).As compared with 30 min after two lung ventilation,PaO2 was significantly elevated (P＜0.05),PaCO2 and PAP were reduced at 15 min after ECMO (P＜0.05).As compared with 15 min after ECMO,PAP was significantly increased at 30 min after pulmonary artery occlusion (P＜0.05).As compared with 30 min after pulmonary artery occlusion,PAP was significantly reduced at 30 min after pulmonary artery open (P ＜ 0.05).ECMO of 3 patients was removed after operation at once and of 15 patients were not removed until 3.3 ± 2.5 days in ICU.Tencases suffered from left cardiac insufficiency to varying degrees after removing ECMO (including 3cases whose ECMO was removed at once after operation),3 of them received ECMO bypass again and 7 of them were relieved after administration of cardiotonic,diuretics and vasodilators.Two cases sufferred from wound fat liquefaction and infection and 2 cases had moderate femoral artery thrombosis after postoperative ECMO.One case had renal failure 5 days after operation and thereafter died of multiple organ failure 2 weeks later after treatment with the continuous ECMO pipelines of hemofiltration,then 17 cases were cured.The pulmonary arterial pressure of all recipients was (30.79 ± 6.18) mmHg 2 months after operation and the cardiac function rating was significantly increased (P ＜0.01).Conclusion The application of ECMO technique in the lung transplantation patients with idiopathic pulmonary hypertension can reduce the pulmonary arterial pressure,improve the safety of operation and has obvious auxiliary effect on the perioperative cardiac function.

Objective Invasive candidiasis is associated with a significant mortality clinically.The purpose of this study was to observe the T cells immune response to fructose bisphosphate aldolase (Fba), an immunodominant antigen of Candida albicans, and determine whether the antigen has the possibility of priming cellular immune protection in invasive candiasis.Methods Using ELISPOT assay, we determined the frequencies of positive spot-forming cells (SFCs) of Fba antigen-specific T cells secreting IFN-γ, IL-4 and IL-17A in the peripheral blood mononuclear cells (PBMCs) of 26 healthy individuals.Results After Fba stimulation, the frequencies of positive SFCs of IFN-γ, IL-4 and IL-17A in the 26 healthy subjects were 23 (9.75, 42.50), 0 (0, 0.25) and 1.5 (0.75, 8.25), respectively, with statistically significant differences among the three (P<0.01).The response rates of IFN-γ (100% [26/26]) and IL-17A (76.92% [20/26]) were significantly higher than that of IL-4 (15.38% [4/26]) (P<0.01).Fba-induced strong response (SCFs ≥20) for IFN-γ was observed in 57.69% (15/26) of the healthy individuals, that for IL-17A in only 1, while that for IL-4 in none.Responses of both Th1 and Th17 cells to Fba were found in 65.38% (17/26) of the subjects, that of Th1 cells in 19.23% (5/26), but that of Th2 cells in none.Conclusion Fba of Candida albicans can induce immunodominant responses of Th1 and Th17 cells and is a potential vaccine against invasive candiasis.

Objective To construct a double luciferase reporter gene vector with PRDM1 gene promoter as target,and establish drug screening cell model in vitro,hope to find small molecule compounds in Chinese herbal medicine library by this model.Methods Total DNA was extracted from 293T cells and it was used for amplifying the fragment contained PRDM1 gene promoter (267 bp/1 257 bp) by PCR.The amplified product was inserted into pGL3-Basic vector.The PCR product and pGL3-PRDM1 vector were verified by sequencing and alignment.The pGL3-PRDM1 and pRL-TK vector were co-transfected into engineer cells.The activity of PRDM1 gene promoter could be assayed by measuring luciferase.The method was optimized by changing ratio of two vectors.Results The highest transfection efficiency and luciferase activity were found in ratio of n(pGL-PRDM1) ∶ n(pRL-TK)=2 ∶1,and with the recombinant luciferase report gene vector contained the length of 1 257 bp amplified fragment transfecting into U266 cells.Moreover,the inductor (5-Aza-CdR) of PRDM1 gene was used for authenticating the method(P＜0.01),the fluoresscence in tensity and promoter activity of PRDM1 gene were enhanced in a dose dependent manner with 5-Aza-CdR.The activity of the promoter of PRDM1 gene was significantly decreased from the concentration of 5 μmol/L of Artemisinid(P＜0.05).The total glucosides of paeoniflorin promoted the promoter activity of PRDM1 gene at a low concentration,and inhibited the promoter activity of PRDM1 gene at a high concentration.Artesunate has no effect on cell proliferation.The effect of total glucosides of paeony on cell proliferation was more complicated.Conclusion A drug selection model targeting PRDM1 gene promoter has been successfully established,and artesunate has been screened to inhibit the promoter activity of PRDM1 gene.

Objective To discuss the effectiveness of anterolateral thigh propeller flaps for treatment of inguinal skin and soft tissue defects.Methods From June,2009 to October,2014,12 patients with inguinal skin and soft tissue defects were treated with anterolateral thigh propeller flaps pedicled with perforator of descending branch of lateral circumflex femoral artery.Of them there were 8 males and 4 females,aged from 22 to 51 years.The left side was involved in 3 cases and right side 9 cases.Defects were caused by traffic accident injury in 5 cases,crash injury of heavy object in 7 cases.There were mere skin and soft tissue in 2 cases,combined with bone fractures,nerves,vessels and muscles injury in 10 cases.The area of defects ranged from 9.0 cm×5.0 cm to 22.0 cm×9.0 cm.During operation,anterolateral thigh propeller flaps ranging from 11.0 cm×7.0 cm to 24.0 cm×1 1.0 cm were used to repair the wounds.Results All flaps and skingrafts survived after operation and the wounds obtained primary healing.After 8-24 months follow-up,all flaps were characterized by soft texture,good color,and satisfactory appearance.According to the Britain's Medical Research Council at 8 months after operation,the sensation of the flaps were recovered to S2 ～ S3+,No obvious scar contracture and chromatosis were observed at donor site.Conclusion Anterolateral thigh propeller flaps pedicled with artery descending branch of lateral circumflex femoral perforator is an ideal choice for the reconstruction for inguinal skin and soft tissue defects.

Objective To investigate the relationships and types between pulmonary subsolid nodules and bronchi and vessels,and their diagnostic values in differentiating subsolid nodules.Methods MSCT images of 40 subsolid nodules were retrospectively reviewed.The relationships between subsolid nodules and bronchi were divided into five types.Type Ⅰ:the bronchi were “cutoff”in the solid part of nodules.Type Ⅱ:the bronchi were distortion and extension in the solid parts of nodules.Type Ⅲ:the bronchi were distortion and extension in the GGO parts of nodules.Type Ⅳ:the bronchi were normal course in the GGO parts of nodules.Type Ⅴ:the bronchi were circumvent nodule lesions.The relationships between subsolid nodules and vessels were categorized into four types.Type Ⅰ:vessels passing by SSNs;Type Ⅱ:intact vessels passing through SSNs;Type Ⅲ:distorted,dilated or tortuous vessels seen within SSNs;Type Ⅳ:more complicated vessels other than described above.The relationship types were correlated to pathologic and/or clinical findings of subsolid nodules.Results Of 40 subsolid nodules,4 were preinvasive nodules,6 micro infiltrating adeocarcinomas and 30 invasive ones that were pathologically proven.Types Ⅰ,Ⅱ,Ⅲ and Ⅳ vascular relationships were observed in 0,8,21 and 11 subsolid nodules,respectively.Type Ⅲ relationship was the dominating one.All 11 subsolid nodules were invasive adenocarcinomas,when the relationship between subsolid nodules and vessle was type Ⅳ.There were 17 invasive adenocarcinomas,2 preinvasive ones,and 2 infiltrating ones when the relationship between subsolid nodules and vessel was type Ⅲ.Correlation analysis showed strong correlation between invasive adenocarcinoma and type Ⅲ and Ⅳ relationships.There was significantly statistical significance among 3 groups of relationships between subsolid nodule and vessels(χ2 =1 5.1 90,P =0.004).Types Ⅰ,Ⅱ,Ⅲ,Ⅳ and V brochi relationships were observed in 20,5,6,9 and 0 subsolid nodules,respectively.Type I relationship was the dominating relationship.There were 1 7 invasive adenocarcinomas, 1 preinvasive one,2 micro invasive ones when the relationship between subsolid nodules and bronchi were typeⅠ.There was significantly statistical significance among 3 groups of relationships between subsolid nodule and bronchi (χ2 =23.81 1,P =0.001 ).Conclusion Different subsolid nodules have different relationships with bronchi and vessels.Understanding and recognizing the characteristic subsolid nodule-bronchi and subsolid nodule-vessel relationships may help to identify which nodules are more likely to be malignant.

Objective To assess the differences in nutrient composition by chemical analysis method and weighing method,A corps of armed police forces daily dietary nutrient intakes were investigated in Tianjin,to understand the status of the actual intakes of dietary nutrient.Methods A dietary survey was performed using the weighing method and with ref-erence to China food composition database in 2002.The intake of each nutrient was calculated through chemical analysis;Nutritional analysis was used in the collection of duplicate food portions during the dietary survey.Results For most of the main dietary nutrients(protein,fat and carbohydrates),minerals (Na,K,Mg,Ca,Zn,Cu and Se)and vitamins (β-car-otene,vitamin E,thiamine,riboflavin,vitamin C,niacin,and vitamin A),the calculated values of intakes were found to exceed those obtained via chemical analysis.Conclusion Differences of the two methods of dietary survey were ultimately due to the analysed values were below the calculated ones.So it is necessary to improve methods of food storage and trans-portation in order to obtain adequate nutrition and to improve the health of troops.