Objective To compare the clinical efficacy of video assisted thoracic surgery using two ports and three ports for pulmonary bullae.Methods Clinical data of 120 patients used video assisted thoracic surgery in the treatment of pulmonary bullae were retrospectively analyzed.62 patients with three ports conventional surgery were selected as three ports group,58 patients with two ports method were selected as two ports group.The two groups were followed up after operation,and the operation time,intraoperative bleeding,extubation time,postoperative drainage volume and hospital stay,the pain scores at postoperative 6h,1 day,3 days,1 week and postoperative complications were compared between the two groups.Results There were no significant differences in operative time and blood loss between the two groups(t=-0.845,-1.164,all P>0.05).After operation,the extubation time[(3.2±1.6)d],postoperative thoracic drainage[(270.8±192.4)mL]and hospitalization time[(5.9±2.1)d] of the two ports group were significantly less than those of the three ports group(t=-4.972,-2.637,-4.601,all P<0.05).The two groups had no postoperative complications,all patients recovered and discharged,followed up,there was no recurrence and other complications.6h,1 day,3 days and 1 week after surgery,the VAS scores of the two ports group were significantly lower than those of the three ports group(t=-5.888,-6.682,-4.190,-5.710,all P<0.01).Conclusion The clinical curative effect of video assisted thoracic surgery using two ports is obvious,the safety is high,and it has high popularization and application value.

Objective To evaluate the diagnostic value of 3-dimentional spiral computer tomography (3 D- sCT) in patients with upper urinary tract obstruction. Methods 113 patients with upper urinary tract obstruction were subjected to 3D-sCT. Results The site of urinary tract obstruction and hydronephrosis were distinctly shown in all patients by 3D-sCT. 112 of them were confirmed by ESWL, pathological or operative findings. Conclusions 3D-sCT can exactly show the location, cause and interaction of the upper urinary obstruction, and is especially applied to patients with resultless IVP and unable to retrograde pyelography.

BACKGROUND:Mesenchymal stem cel s as potential seeded cel s have been widely used in tissue engineering and clinic therapy;thus, the precise, safe, effective isolation of mesenchymal stem cel s is the particular important premise to build culture system.
OBJECTIVE:To review the methods of isolating mesenchymal stem cel s and to compare the merit and demerit of different methods, thereby providing theoretical basis for safe and high-effective isolation of mesenchymal stem cel s.
METHODS:A computer-based online research of CNKI and PubMed databases was performed to col ect articles, which included reviews, clinical trials and experiments, published between 1965 and 2014 with the key words of“mesenchymal stem cel s (MSCs), isolation methods”in Chinese and English. A total of 52 articles were included according inclusion and exclusion criteria
RESULTS AND CONCLUSION:(1) The whole bone marrow culture method can derive a mass of mesenchymal stem cel s, which need to be purified. (2) The density gradient centrifugation method which uses the media with the density of 1.073 g/mL can be used to harvest more purified cel s. (3) The tissue digestion method is suitable for digestion and isolation of adipose tissue and umbilical cord tissue. Type II col agenase digestion is better, but they are both limited by a high demand for operative techniques. (4) Immunomagnetic bead separation is appropriate to study the biological characteristics of a kind of subpopulation of mesenchymal stem cel s which express special surface markers. (5) The combination method is also an optimal way. (6) Some new methods limited by few dates require further studies.

BACKGROUND:In previous studies, it is satisfied to sorting bone marrow mesenchymal stem cels based on natural sedimentation combined with low permeation. Then, based on particle hydromechanics theory, the settling velocity of bone marrow mesenchymal stem cels in culture liquid is calculated. A simple and easy method of separation, purification and proliferation of bone marrow mesenchymal stem cels is established by natural sedimentation velocity. OBJECTIVE:To explore the feasibility of sorting bone marrow mesenchymal stem cels by natural sedimentation velocity method. METHODS:The density interval of rabbit bone marrow mesenchymal stemcels (ρ1) was determined by density gradient centrifugation method. The diameter of rabbit bone marrow mesenchymal stem cels (d) was measured by scanning electron microscope. The density of culture liquid (ρ2) was measured by liquid density meter. The viscosity of the culture liquid (μ) was measured by viscosity meter. The settling velocity of bone marrow mesenchymal stem cels (Vt) was derived from the above four numerical values with the appropriate formula. Bone marrow mesenchymal stem cels were sorted from bone marrow tissue by natural sedimentation velocity method. Cel proliferation, purity and differentiation were observed. Meanwhile, the primary culture time of three different cel sorting methods was recorded; the colony formation rate of rabbit bone marrow mesenchymal stem cels was determined. RESULTS AND CONCLUSION:(1) The diameter of rabbit bone marrow mesenchymal stem cels was (20.37±4.58) μm, and the setting velocity of rabbit bone marrow mesenchymal stem cels in the culture liquid was 50-55 mm/h. (2) Bone marrow mesenchymal stem cels could be successfuly isolated from bone marrow tissue of rabbits by the natural sedimentation velocity method, which could be induced into osteoblasts and adipocytes. (3) The natural sedimentation velocity group cost less time than the other two density gradient centrifugation groups in the primary culture stage. The colony formation rate of the natural sedimentation velocity group was higher than that of the other two groups. (4) Natural sedimentation velocity method did not impose any intervention measures for sorting cels, which maybe maximaly maintain cel viability and biological characteristics. The whole separation process was simple and safe, which may have little damage to the cels.

BACKGROUND:Thoracic vertebra connected to the corresponding section of the ribs constitute thoracic vertebral, which is deep, and the structure is complicated, so it is difficult to ful y expose thoracic vertebrae. Usual y, corresponding ribs is removed, and the injured site wil be reached through thoracic cavity. The trauma is big. Some complications often occur such as chest pain, and local skin numbness. Therefore, whether it is possible to reach the same target without removal of ribs through intercostal space became a new clinical problem. OBJECTIVE:To explore the safety and efficacy of autologous iliac bone graft and plate fixation for tuberculosis of thoracic vertebra. METHODS:A total of 30 patients diagnosed with tuberculosis of thoracic vertebra from January 2008 to December 2013 were conventional y treated with anti-tuberculosis treatment for 2 to 3 weeks, and then treated with autologous iliac bone fusion through intercostal space and anterior plate fixation. Postoperative fol ow-up was conducted from 6 to 22 months. Fracture healing condition, the degree of pain relief, Cobb angle change, length of incision, blood loss, operation time, postoperative recovery of neurological function were observed. RESULTS AND CONCLUSION:In 30 patients, the length of incision was (12.4±1.8) cm;longitudinal incision distraction width was (10±3.2) cm;the time of opening the chest was (16.0±2 .5) minutes;the time of closing the chest was (12.0±1.5) minutes;intraoperative blood loss amount was (430.0±87.4) mL. Preoperative and postoperative average kyphosis angles were respectively 27° and 8°, with an average rectification of 19°. The pain basical y relieved at 1 to 2 weeks after the surgery. 28 patients were healed, and the symptoms of 2 patients were improved. Postoperative fol ow-up radiographs revealed that autologous bone grafts were thoroughly fused, and the fusion time lasted from 4 to 5 months. These data verified that autologous iliac bone graft through intercostal space and plate fixation is an effective, safe method for tuberculosis of thoracic vertebra. The exposed range through intercostal space can satisfy the operation requirements of complete tuberculosis clearance, autologous iliac bone graft and plate fixation, and can ensure the integrity of the whole thorax and spine stability.

Objective:To explore the damage of the intestinal mucosal barrier of septic rats by the activation of NOD-like receptor family, pyrin domain-containing 3 (NLRP3) inflammasomes and the role of Ulinastatin (UTI) on the expression of intestinal nuclear factor-κB (NF-κB)/NLRP3 inflammasome signaling pathway in septic rats.Methods:According to the random number table method, 64 male Wistar rats were divided into sham operation group (Sham group), cecal ligation and puncture (CLP) group, UTI treatment group (100 kU/kg UTI was intraperitoneally injected 1, 6, 12 and 18 hours after CLP), and UTI pretreatment group (100 kU/kg UTI was given 1 hour before CLP), with 16 rats in each group. The survival of rats was observed after 24 hours, and the blood was collected from abdominal aorta at 24 hours after modeling, then rats were killed and their ileum tissues were taken. Hematoxylin-eosin (HE) staining was used to observe histopathological changes and Chiu score. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and intestinal fatty acid binding protein (I-FABP) in serum were detected by enzyme linked immunosorbent assay (ELISA). The protein expression of NF-κB p65 in intestinal tissue was detected by Western blotting. The expression of intestinal tight junction proteins Claudin-1, Occludin and the inflammasome NLRP3, apoptosis-associated speck-like protein containing CARD (ASC) and caspase-1 were detected by immunohistochemistry.Results:Compared with Sham group, the 24-hour survival rate of CLP group was significantly reduced. Histopathological results showed that the CLP group had severe edema of mucosa and submucosal stroma with obvious infiltration of inflammatory cells and disordered villi arrangement. Some glands were incomplete, and the villus structure was severely damaged. The Chiu score was significantly increased. The levels of TNF-α, IL-1β, I-FABP in serum and the protein expression of NF-κB p65 in intestinal tissue were significantly increased. The positive expressions of NLRP3, caspase-1 and ASC were also significantly increased. However, the positive expression of tight junction protein in small intestine tissue such as Occludin and Claudin-1 were significantly reduced. It suggested that when sepsis occurs, small intestinal mucosal barrier dysfunction happens, and mucosal permeability increases, while tight junction protein expression decreases, NLRP3 inflammasome and its upstream molecule NF-κB p65 were activated. After UTI treatment and UTI pretreatment intervention, although there was no significant difference in 24-hour survival compared with CLP group (62.5%, 68.8% vs. 43.8%, both P > 0.05), the intestinal tissue damage of septic rats was significantly improved. Specifically: Chiu score and the levels of TNF-α, IL-1β, I-FABP in serum were significantly decreased [Chiu score: 3.37±0.25, 3.23±0.16 vs. 4.08±0.13, TNF-α (ng/L): 147.62±20.74, 140.71±24.81 vs. 222.82±16.84, IL-1β (ng/L): 80.64±5.68, 78.11±4.75 vs. 133.73±3.92, I-FABP (μg/L): 38.29±3.60, 35.88±4.52 vs. 59.81±4.66, all P < 0.05]; the protein expression of NF-κB p65 was significantly decreased (NF-κB p65/β-actin: 0.65±0.10, 0.69±0.11 vs. 0.99±0.10, both P < 0.05), the positive expressions of Claudin-1 and Occludin in the small intestine tissue were increased [Claudin-1 positive expression area: (19.43±3.08)%, (23.99±6.27)% vs. (7.77±2.03)%; Occludin positive expression area: (19.58±4.75)%, (23.28±3.68)% vs. (11.69±4.30)%, all P < 0.05], while the positive expressions of NLRP3, caspase-1, ASC were decreased [NLRP3 positive expression area: (7.80±3.14)%, (6.86±2.63)% vs. (14.44±3.68)%; caspase-1 positive expression area: (10.62±3.52)%, (9.49±3.09)% vs. (26.69±8.05)%; ASC positive expression area: (9.95±2.81)%, (10.53±3.61)% vs. (24.16±5.48)%, all P < 0.05]. However, there was no significant difference in the improvement effect between UTI treatment group and UTI pretreatment group.Conclusions:Intestinal barrier dysfunction in sepsis may be related to the activation of NLRP3 inflammasomes in the intestinal mucosa. The protective effect of UTI in the intestinal mucosa may be related to inhibiting the activation of NLRP3 inflammasomes in the intestinal mucosa, but UTI pretreatment has no obvious advantage compared with UTI treatment.

Objective To present a medical image transmission scheme based on fast healthcare interoperability resources(FHIR) proxy gateway which can enable medical personnel to access the hospital's medical imaging system via the Internet using mobile terminals,and then raise the medical staffs' working efficiency.Methods RESTful WebServices as the interoperability mechanism of image data was used in combination with FHIR image resource model and construct an intermediate gateway to three-tier network architecture,in order to solve the problem of transmitting image data to mobile terminals through gateway via the hospital's network.Results During its half-year trial run,the Internet mobile terminal reading system based on this method ran stably and was in good condition,in the actual hospital environment.Conclusion The gateway method is simple,flexible,and can fully support the mobile terminals' access to the background image data center.

Objective To investigate the antitumor effect of Haishengsu injection extracted from marine shellfish. Methods Transplant tumor models of sarcoma 180 (S 180), Ehrlich ascites carcinoma (EAC), and hepatoma (Heps) in mice were established. Different doses of Haishengsu injection were given to the mice and the tumor inhibition rates of Haishengsu injection, life span of the mice were calculated. Results The tumor inhibition rates of haishengsu injection (490～1000mg?kg -1?d -1,iv) were 41.10%～49.08% in mice with S 180 and 36.29%～49.19% in mice with hepatoma,respectively. The same doses of Haishengsu injection prolonged the life spans of EAC-bearing mice by 22.93%～69.98%. Conclusion The haishengsu injection has the antitumor effects on the tumor-bearing mice without evident side effects.

Background Pluripotent stem cell-derived retinal pigment epithelial (RPE) cells holds great promise for the treatment of age-related macular degeneration (AMD) and retinitis pigmentosa (RP),but the poor induction efficiency and the according high cost of RPE differentiation hindere its clinical applications.Curcumin is proved to have a promoting effect on the induced differentiation of embryonic stem cells (ESCs).However,the mechanism of curcumin on differentiation of human ESCs into RPE-like cells remains unclear.Objective This study aimed to explore the underlying molecular mechanism of curcumin on directed differentiation of human ESCs into RPE-like cells.Methods Human ESCs strains were cultured in the Matrigel-coated 6-well plate with mTeSRTM 1 medium until over-confluence,and basic fibroblast growth factor was withdrawn there after to induce automatic differentiation.Curcumin at the final concentration 1 μmol/L was added in the first day of differentiation for 24 hours,and the cells without curcumin in the medium served as the control group.Total RNA and protein were extracted at 3 weeks and 5 weeks after induction.RT-PCR,Western blot and immunofluorescence were performed to examine the expressions of the biomarks of stem cells and RPE cells as well as Wnt/β-catenin signaling pathway components.The endocytosis of polystyrene microsphere by induced RPE (iRPE) cells was investigated to verify their function of phagocytosis which features RPE cells.Results Pigmented cells were found from 3 weeks through 5 weeks after induction in the curcumin group,but only less pigmented cells were seen in the fifth week after induction in the control group.In the third and fifth week after induction,the relative expression levels of NANOG mRNA in the iRPE cells were significantly lower than those in the control group (t =13.086,P =0.022;t =34.186,P =0.004),and the relative expression levels of Pax6,RX,CRALBP and RPE65 mRNA were higher in the curcumin group than those of the control group (all at P＜0.01).Western blot assay showed that the expressing bands for CRALBP,RPE65 and MITF enhanced in iRPE cells with a similar appearance in human RPE cells.However,these expressions were all absent in human ESCs.Immunofluorescence staining showed the positive expressions of Pax6,MITF and ZO-1 in cytoplasm of iRPE cells in the curcumin group with a purified efficacy 100％.The fluorescence dye-doped polystyrene microspheres in cytoplasm were obvious in the iRPE cells like positive controls,but the polystyrene microsphere was absent in the negative controls.From 3 weeks through 5 weeks after induced,the relative expression levels of Lef1,MYC and TCF7 mRNA (the dwnstream target genes of Wnt signaling pathway),FZD3 mRNA (Wnt receptor),Wnt2B mRNA (Wnt ligand) and Wnt7B mRNA were significantly reduced in the curcumin group compared with the control group (all at P＜0.01).Conclusions Curcumin promotes the differentiation of human ESCs into RPE-like cells by stimulating the activation of Wnt signaling pathway,and therefore accelerate the differentiation and mature of iRPE cells.

A new approach based on online coupled liquid chromatography-gas chromatography-mass spectrometry ( LC-GC / MS ) was developed for the rapid determination of 4-( methylnitrosamino )-1-( 3-pyridyl)-1-butanone (NNK) in mainstream cigarette smoke, in which a switching valve was employed to online switching between two-dimensional chromatography. The online LC-GC / MS system used in this study was built by using online gel permeation chromatography-gas chromatography-mass spectrometry except that the micro gel column was replaced by micro alkaline alumina column which was prepared by ourselves before. The NNK in mainstream cigarette smoke was collected by a Cambridge filter pad, then the pad was extracted with dichloromethane, and the extract was quantitatively analyzed by online LC-GC / MS with D4-NNK as an internal standard. Online LC-GC / MS allowed online pretreatment purification, and the sample was subjected to online LC-GC / MS without any prior purification, which reduced human error in analysis process. The injection volume of the present online LC-GC / MS could reach 40 μL, which was 20 times of that of the conventional GC / MS (2. 0 μL of injection volume), and thus significantly improved the sensitivity. Under the optimum conditions, good linearity ( r = 0. 9998) was obtained over the range of 1. 2 - 120 ng / mL. The recoveries at three spiked levels ranged from 93. 9% to 96. 0% , and the limits of detection for qualitative and quantitative detection were 0. 25 ng / mL and 0. 9 ng / mL, respectively. All the results obtained by the present method are comparable to those of standard method recommend by China National Tobacco Company.