BACKGROUND:Periosteum is considered as a source of seed cels for cel therapydue toits biological features. OBJECTIVE:To seek the optimal way to isolate and culture rabbit periosteal cels and identify their biological features. METHODS:Rabbit periosteum on facies medialis tibiae was taken out under aseptic conditions. Periosteal cels isolated through the digestion of type II colagenase with the explants culture method were cultured in DMEM/F12 complete medium. Cel ultrastructure was observedunderan inverted microscope. Periosteal cel proliferation was determined bycel counting kit-8assay. Cel surface antigensCD90 and CD105 were determined using flow cytometry. Osteogenic andlipogenic induction mediums were applied to induce periosteal cels to differentiate into osteocytes and adipocytes, respectively. After 2 weeks of induction, cels were harvestedfor alizarin red staining and oil red O staining to assay the calciumnodules and lipid droplet. RESULTS AND CONCLUSION:The digestion of type II colagenase with the explants culture method shortened the period of primary cels culture and enhanced the survival rate, which causedhigher purity and stronger reproductive activity of harvested periosteal cels. Primary cultured periosteal cels grew in form of spindle spiral or paralel. Alizarin red andOil red O staining verified the multi-directional differentiation potentiality of periosteal cels. These findings suggest that the periosteal cels with high purity,strong reproductive activity,andmulti-directional differentiation potentialitycanbe harvested in short time using digestion of type II colagenase with the explants culture method.

Background and purpose:Visceral pleural invasion (VPI) and vessel invasion (VI) are poor prognostic factors in patients with non-small cell lung cancer (NSCLC). The primary initial recurrence site may be local recurrence in VPI and distant metastasis in VI. The purpose of this study was to validate the prognostic impact and effect of the initial recurrence site of VPI and VI on survival outcomes for NSCLC.Methods:Two hundred and ninety patients who were diagnosed as having NSCLC and underwent lobectomy between Jan. 2007 and Dec. 2013 were retrospectively analyzed. VPI was identiifed in 51 patients as VPI group, the other 239 patients without VPI as non-VPI group. VI was identiifed in 29 patients as VI group, the other 261 patients without VI as non-VI group. Clinical characteristics, overall survival (OS), disease-free survival (DFS) were compared.Results:There were statistically signiifcant differences between VPI group and non-VPI group in tumor size, lymph node metastasis, TNM stage and initial recurrence site (P<0.05). Furthermore, there were statistically signiifcant differences between VI group and non-VI group in lymph node metastasis and TNM stage (P<0.05). The 1-, 3- and 5-year OS rates in VPI group (88.2%, 56.7% and 52.7%) were lower than those in non-VPI group (95.8%, 83.7% and 74.0%,P<0.001). The 1-, 3- and 5-year OS rates in VI group (79.3%, 56.8% and 48.7%) were lower than those in non-VI group (96.1%, 81.3% and
72.3%,P=0.001). Cox regression showed TNM stage was a significant prognostic factor for DFS, whereas lymph node metastasis and VPI were signiifcant prognostic factors in patients with NSCLC.Conclusion:The primary initial recurrence site in VPI patients is local recurrence. Patients with VPI or VI may need more postoperative therapy because of their poor prognosis.

Objective An incentive system was established to promote the rapid development of scientific research and to improve the scientific management ability.Methods The research incentive system, which including research awards, research fund management, research fund matching, and performance management, was implemented in our hospital, aimed to develop a clear quantitative assessment index to encourage research activity, and maximize the research outcome.Results It is clear that the research project application and output in output have been increased;and it positively associated with the number of years of implementation with the statistical significance.Conclusions Establishment of the research incentive system has encouraged the clinicians to make effort to conduct research projects which has largely increased the number of research projects and produced better outcomes.Applying such system has enhanced the capacity of core research of the hospital.

Objective To investigate the expression of cyclin G1、cyclin G2 in gastric carcinoma and its significance. Methods The mRNA expression of cyclin G1、cyclin G2 in 55 cases of gastric carcinoma was measured with RT-PCR. The protein expression of cyclin G1 and cell cycle were detected by flow cytometry.Results Expression rate of cyclin G1 in gastric carcinoma was 58%,which was higher than that in normal tissue(P

Objective: To investigate the influence and significance of DHA on expression of angiogenesis-related genes in SGC7901 cells. Methods:SGC7901 were treated with DHA (5, 10, 20, 40, and 80μmol/l) for different times (24, 48, and 72 h), and the growth inhibition was detected by MTT. The expression of vascular endothelial growth factor (VEGF-C), cyclooxygenase-2 (COX-2) vascular cell adhesion molecule-1 (VCAM-1), and PTEN mRNA were detected by fluorescence-based quantitative poly-merase chain reaction (qRT-PCR). Their corresponding protein levels were tested by Western blot. Results:DHA significantly inhibited the growth of SGC7901 cells in a dose-and time-dependent manner (P<0.05). The expression of the angiogenesis-related genes signifi-cantly changed, as shown by RT-PCR and Western blot analyses. Compared with the control group, the expressions of VEGF-C, COX-2, and VCAM-1 were down-regulated, whereas the expressions of PTEN were up-regulated, after DHA treatment (P<0.05). Con-clusion:DHA inhibits cell growth in gastric cancer SGC7901 cells. The effect may be due to its reduction of VEGF-C, COX-2, and VCAM-1 gene expression, as well as its promotion of PTEN expression in gastric cancer cells.

BACKGROUND:The reported time of bone marrow mesenchymal stem cel s induced to differentiate into chondrocytes is different. Few studies have observed and compared the cel s’ dynamic transformation during the induction process.
OBJECTIVE:To observe the dynamic differentiation and the mature time of rabbit bone marrow mesenchymal stem cel s which were directional y induced to chondroblasts for 8, 11, 14, 17, 20 days.
METHODS:Bone marrow was aspirated from the femur of New Zeal rabbits, and bone marrow mesenchymal stem cel s were isolated by gradient centrifugation. After cultivation and amplification, bone marrow mesenchymal stem cel s at passage 3 were directional y induced to chondrocytes by the serum-free medium containing transforming growth factor beta-1. The experiments were divided into five groups according to different induction time points:8 days, 11 days, 14 days, 17 days, 20 days. Then cel ular morphology, toluidine blue staining, typeⅡ col agen immunohistochemistry, aggrecan content in induction medium, and chondrogenic differentiation in each group were observed and compared.
RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cel s had apparently transformed in morphology at 8 days of induction, and presented obvious chondrocytes’ morphology at 14 days. The aggrecan in induction medium could be detected at a low level at 4 days, significantly increased at 8 days, and maintained slow increasing at 20 days. At 14 days, the metachromatic particles could be found by toluidine blue staining, and the col agen type Ⅱimmunohistochemistry was significantly positive in cel climbing slice. Experimental findings indicate that, bone marrow mesenchymal stem cel s that are monolayer cultured in a high density can be induced into chondroblasts at the effect of transforming growth factor beta-1 and other factors. There are a few chondroblasts in the early induction process, then cel s begin to have chondrocytes morphology and function after induced for 8 days, and may differentiate to mature chondrocytes at 14 days. In addition, they can keep a high biological activity in the induction process.

Objective To investigate the effects of dihydroartemisinin (DHA) on the proliferation of gastric cancer cell line SGC7901 and its mechanism.Methods SGC7901 cells were divided into the DHA group and the control group.SGC7901 cells in the DHA group were treated with DHA of different concentrations (6.25,12.50,25.00,50.00,100.00 μmol/L),SGC7901 cells in the control group were cultured in the 0.1％ DMSO medium.The proliferation of SGC7901 cells was detected by the MTF method at different time points (24,48,72 hours).Cell cycles of SGC7901 in the DHA group were observed by flow cytometry at 24 hours after treatment.The expressions of Cyclin A,Cyclin D1,Cyclin E,Cyclin-dependent kinase 4 (CDK4) and P16 were detected by Western blot after treating SGC7901 with DHA at concentration of 100μmol/L for 24 hours.The interaction between CDK4 with Cyclin D1 or P16 was examined using the co-immunoprecipitation assay.All data were analyzed using the one-way analysis of variance or the t test.Results The proliferation of SGC7901 cells was significantly inhibited after the treatment with DHA at different concentrations (6.25,12.50,25.00,50.00,100.00 μmol/L) for 24,48 and 72 hours (F =78.66,235.37,93.75,P ＜ 0.05).Compared with control group,the number of SCG7901 cells in the G0/G1 phase in the DHA group was significantly increased (F =18.42,P ＜0.05).After treating SGC7901 cells with DHA for 24 hours,the protein expressions of Cyclin D1 and CDK4 were 0.67 ± 0.15 and 0.64 ± 0.18 in the control group,which were significantly higher than 0.17 ± 0.05and 0.24 ± 0.06 in the DHA group (t =7.746,5.164,P ＜ 0.05).The protein expressions of Cyclin E were 0.42 ± 0.06 in the control group and 0.35 ± 0.06 in the DHA group,with no significant difference (t =2.021,P ＞ 0.05).The protein expressions of Cyclin A were 0.35 ± 0.09 in the control group and 0.38 ± 0.08 in the DHA group,with no significant difference between the 2 groups (t =1.266,P ＞ 0.05).The protein expressions of P16 were 0.29 ± 0.07 in the control group and 0.54 ± 0.12 in the DHA group,with significant difference between the 2 groups (t =4.408,P ＜ 0.05).The results of co-immunoprecipitation assay showed that DHA decreased the interaction between CDK4 and Cyclin D1,and increased the interaction between CDK4 and P16.Conclusion DHA induces SGC7901 cells arrested in G0/G1 phase,and the effect may be related with its downregulation of Cyclin D1 and CDK4,up-regulation of P16,decreasing the interaction between CDK4 and Cyclin D1,and increasing the interaction between CDK4 and P16.

Objective To investigate the possible mechanism of glomerular injury in diabetes mellitus by determining whether epithelial-mesenchymal transition (EMT) is caused by high glucose in mice podocytes. Methods Using mice glomerular podocyte cell line as an in vitro system, podocytes were incubated with glucose(12.5 mmol/L, 25 mmol/L, 50 mmol/L) and mannitol (50 mmol/L) for 36 hours. Then the cells were collected and expression of alpha-smooth muscle actin(α-SMA), fibronectin (FN), CD2 associated protein (CD2AP) and Wilms' tumor 1 gene (WT-1) was detected by Western blot and indirect immunofluorescence staining. Results Under low glucose (5.6 mmol/L) and mannitol (50 mmol/L) condition, there were high expression of CD2AP and WT-1, and low expression of α-SMA and FN in mice podocytes. After 36 hours treatment with high glucose (12.5 mmol/L), the expression of α-SMA and FN in podocytes was significantly increased, and the expression of α-SMA and FN was further up-regulated with the increase of glucose dosage (25, 50 mmol/L). The indirect immunofluorescence staining revealed the similar result, and the percentage of positive α-SMA cells was also increased compared with low glucose and mannital group (P<0.05). Meanwhile, Western blot showed that high glucose could down-regulate the expressions of CD2AP and WT-1 in a dose-dependent manner. Conclusion EMT may be a potential pathway leading to podocyte dysfunction and glomerular injury under high glucose conditions.

Objective To improve lesion detection of the radial head with adjusted elbow joint lateral position on X-ray.Methods (1)20 subjects underwent elbow joint CT three-dimensional reconstruction (average intensity projection),which was as X-ray radio-graphy image.The tilt angle of the humerus and the projection direction for fully display of the radial head in projection images and the tilt angle of the humerus against detector in X-ray radiography were measured.(2)20 subjects (patients and volunteers)with el-bow joint disease underwent routine and improved elbow joint lateral position were enrolled.And assessed the bare blocking rate of the radial head articular surface of the two positions.Results (1)The angle between the the humerus and the direction of projection were 49°-60°and 30°-41°in CT and X-ray,respectively.The average angle was (35.5±1)°.(2).The blocking rate and bare bloc-king rate of the radial head articular surface in routine elbow joint lateral position were 71.6%-100% and 0%-28.4%,respective-ly.The average rate of bare blocking was 14.2%.Fracture 13 cases and 5 cases of suspected fractures.The blocking rate and bare blocking rate of the radial head articular surface in improved elbow joint lateral position were 4.8%-25% and 75%-95.2%,re-spectively.The average rate of bare blocking was 85.1%.Fracture 1 7 cases and 1 case of suspectet fractures.Conclusion The im-proved elbow joint lateral position of X-ray can display the radial head articular surface better than the routine position.

BACKGROUND:Periosteal cel s have been used in bone repair, but whether nucleus puplousus cel s co-cultured with autologous periosteal cel s can differentiate into osteoblasts in spinal fusion is rarely reported. OBJECTIVE:To isolate nucleus puplousus cel s and periosteal cel s so as to observe the osteogenic ability of nucleus puplousus cel s co-cultured with periosteal cel s or not. METHODS:Type II col agenase digestion method was used to isolate and purify nucleus pulposus cel s, which were confirmed by toluidine blue and immunohistochemical staining. Periosteal cel s were isolated histological y and cultured in complete medium, and cel surface antigens CD90, CD105 were identified by immunofluorescence staining. According to the experimental needs, the cel s were assigned into two groups. Nucleus pulposus cel s and periosteal cel s were co-cultured by osteogenic induction medium in the experimental group. Nucleus pulposus cel s in the control group were cultured alone in osteogenic induction medium. Cel morphology was observed by inverted microscopy, and cel proliferation was detected by cel counting kit-8. The osteogenic differentiation indexes of cel s in each group were measured using alkaline phosphatase staining, alizarin red staining, and type I col agen immunohistoehemical staining. The expression of osteopontin was tested by western blot assay. RESULTS AND CONCLUSION:CD105 and CD90 expressions of the periosteal cel s were positive. Nucleus puplousus cel s were positive for toluidine blue and col agen type II immunohistochemical staining. The proliferative ability of nucleus puplousus cel s was significantly higher in the experimental group than the control group at days 1, 3, 5, 7, 9. After 2 weeks of induction, the cel s were positive for alkaline phosphatase staining, alizarin red staining, and type I col agen immunohistoehemical staining, but the experimental group showed higher positive expressions than the control group (P<0.05). The expression of osteopontin was also higher in the experimental group than the control group. These findings indicate that nucleus puplousus cel s possess osteogenic ability, but have lower proliferative ability in vitro. After co-culture with periosteal cel s, the proliferative ability of nucleus puplousus cel s can be increased. Under osteogenic induction, nucleus puplousus cel s co-cultured with periosteal cel s have good compatibility and adhere with each other, which have stronger osteogenic ability than cel s cultured alone.