Objective To purify BMSCs by a simple Colony-Forming system and identify BMSCs in vitro.Methods BMSCs were planted at low-density ( 10/cm2 ) in the early stage of isolation,additional cells were scraped until a colony formed,secondly seeded those Colony-Forming cells at low-density.This process was repeat again when passaged cells formed another colony.This so called Colony-Forming selection was repeated several times until highly purified cells were obtained.Biological characters of BMSCs were observed.The surface antigens of BMSCs were identified by flow cytometry.The multipotentiality of BMSCs was assayed for differentiation into either osteoblasts or adipocytes.Results Homogeneous cells were obtained after BMSCs were passaged twice to thrice by Colony-Forming system.These BMSCs highly expressed positive surface antigens of BMSCs ( CD29 in 98.8％,CD90 in 98.4％ ) meanwhile seldom expressed negative surface antigens of BMSCs (CD31 in 2.6％,CD45 in 3％ ).BMSCs efficiently differentiated into osteoblasts and adipocytes.Conclusions Colony-Forming system is a simple and effective way to get highly purified BMSCs in a short time.

BACKGROUND: Previous studies have confirmed that in the animal models of articular cartilage defects and osteoarthritis, the chondrocytes can overexpress the matrix metal oproteinases. Various abnormal stimuli are likely to break the balance between matrix metal oproteinase and tissue inhibitor of metal oproteinase, thus leading to degeneration of extracel ular matrix of articular cartilage, as wel as the decline and offset of cartilage chondrocytes. OBJECTIVE: To observe the effect of cyclic tensile strain on the expression of matrix metal oproteinases during the repairing process of rabbit articular cartilage defects. METHODS: The animal models of articular cartilage defects were established, and chondrocytes were separated for culture at 10 weeks after operation. The chondrocytes on the non-surgical side were considered as the normal group, and the chondrocytes on the surgical side were randomly divided into high cyclic tensile strain group, low cyclic tensile strains group and control group, and the load amplitude was sin10%. Then 0.1, 1.0 and 0 Hz cyclic tensile strains were loaded respectively. The expressions of matrix metal oproteinases 2, 3, 9 and 13 in each group were detected with reverse transcription-PCR at 24, 48 hours, 1, 2 and 4 weeks after loading cyclic tensile strain. RESULTS AND CONCLUSION: There were significant differences in the expressions of matrix metal oproteinases 2, 3, 9 and 13 at 24 hours after loading cyclic tensile strain between the normal group and the control group (P < 0.05); and there were significant differences in the expressions between the high cyclic tensile strain group and the low cyclic tensile strain group at 1, 2 and 4 weeks after loading cyclic tensile strain (P < 0.05).At the same time, the expressions of matrix metal oproteinases 2, 3, 9 and 13 in the low cyclic tensile strain group were continued to decline, and there were significant differences in the expressions after loading cyclic tensile strain for 24 hours and 4 weeks (P < 0.05). The results indicate that mechanical load can affect the expression of matrix metal oproteinases in the healing process of rabbit articular cartilage defects. In the cel ular and molecular level, the incidence and development of pathological articular cartilage defect and stress should affect each other.

Objective To observe the biocompatibility of a biomimetic designing of a multi-grade compositions in repairing articular cartilage and subchondral bone in animal bodies and repair the fullthickness defects in articular cartilage with the compositions and to study the regenerated cartilage histomorphologically. Methods Biocompatibility study: Acute general toxicity test, Haemolysis test, subcutaneous implantation test and chronic toxicity test. Articular cartilage defects repaired experimental study :The models of defects in articular cartilage were made artificially in both condylus lateralis femoris of mature rabbits. Implanted with the biomimetic designing of a multi grade compositions randomly at one side as the experimental group and the other side were untreated as the control group. The rabbits were killed at 4, 6, 8and 12 weeks after operation, respectively, with 6 ones at each time, and the macroscopic, histological, ultrastroctural examinations and semi-quantity cartilage scoring employing Wakitanifa repaired cartilage value system were performed. Results Biocompatibility study: (1) The rabbits' weight in experimental group kept growing .(2) Haemolysis rate of rats to different concentrations of diffusion solution was＜5%.(3) In chronic toxic reaction, rabbits' liver and kidney function was not different compared with the control groups at 12weeks and the index before operation. Articular cartilage defects repaired experimental study: 4-8 weeks after operation, the defects in the experimental group were partly filled with hyaline cartilage. Twelve weeks after operation, the defects in the experimental group were completely filled with mature hyaline cartilage.However, fibrous tissues were seen in the control group all the time. At 4, 6, 8, and 12 weeks postoperatively, the Wakitanifa cartilage scores were (7.60±0.98), (5.69±0.58), (4.46±0.85) and (4.35±0.12), respectively,in the experimental group and (10.25±1.05), (9.04±0.96), (8.96±0.88) and (8.88±0.68), respectively, in the control group. Differences between the control group and the experimental group were significant. Conclu sion The biomimetic designing of a multi-grade compositions has good biocompatibility and may induce cartilage regeneration to repair the full-hickness defects of articular cartilage.

Objective To investigate the biological mechanisms and feasibility of collagen/hydroxyapatite scaffolds for repairing articular cartilage and subchondral bone defects. Methods 45 New Zealand white rabbits were included. Full-thickness articular cartilage defect models were established in experiment and control groups and the biomaterial scaffolds were implanted in the experiment group. The animals were sacrificed in the 4th, 6th and 8th week after the operation respectively. Histological scores were performed by semi-quantity cartilage scoring system and the expressions of matrix metalloproteinases (MMPs)-2, 3, 9 and 13 mRNA in the synovialis were detected with RT-PCR. Results The repaired tissues were totally hyaline cartilages in the experiment group 2 weeks after operation and there were no repaired tissues in the control group. There were significant differences between these 2 groups in Wakitani scores (P<0.01). The activation and expression of MMP-2, 3, 9 decreased in the experiment group (P<0.05), but the activation of MMP-13 sustained and the expression of MMP-13 could be detected during the observed period (P>0.05). Conclusion The new biological scaffold can alleviate the inflammatory and protect the cartilages by inhibiting the expression of MMPS, which promote the reconstruction of articular cartilage in rabbits.

OBJECTIVE: To assess the analgesic efficacy of compound lidocaine cream in lumbar puncture of children with leukemia and lymphoma. METHODS: 312 leukemia and lymphoma children necessarily undergone lumbar puncture were divided into compound lidocaine cream (cream) group and compound lidocaine injection (injation) group as control with 156 cases respectively according to the will of inpatient children and their family. For cream group, compound lidocaine cream was smeared on the skin around the lumbar puncture point evenly and covered with sterile and transparent dressing for about one hour before lumbar puncture. Then the cream and transparent dressing were removed and lumbar puncture was performed after regular disinfection. For control group, 2% compound lidocaine was injected as local anesthesia before lumbar puncture. The extent of pain was evaluated by Wong-Baker Faces Pain Rating Scale and FLACC Scale as well as children's physiological indexes (heart rate, breathing, blood pressure). The lumbar puncture success rate and the discomfortableness as well as family satisfaction were recorded by special person. RESULTS: Compared with control group, the incidence of pain and discomfortableness in cream group significantly reduced, and the family satisfaction significantly increased (P＜0.05). There were obviously differences in the physiological indexes (heart rate, breathing, systolic blood pressure) before and after lumbar puncture in control group (P＜0.05). However, the physiological indexes in cream group had no obvious change. There was no difference on the success rate of lumbar puncture between the two methods of anesthesia. CONCLUSION Compound lidocaine cream has significantly analgesic effect which can relieving pain caused by lumbar puncture for children.
Analgesics ; Anesthetics, Local ; Child ; Hematologic Neoplasms ; Humans ; Lidocaine ; Lidocaine, Prilocaine Drug Combination ; Spinal Puncture

ObjectiveThe purpose of this study was to observe the correlation of mTOR and VEGF gene with nephropathy indicators in diabetic rats. Methods Forty-eight male Sprague-Dawley(SD)rats were divided into diabetes mellitus group(DM=28)and control group(NDM=20). Diabetic models were produced by injection of streptozotocin. In the courses of 12,16,20 and 24 weeks,the histology scores(HS)and glomerular basement membrane(GBM)thickness were collected. The protein and mRNA expressions of the gene of mTOR,VEGF and VEGFR2 were observed by immunohistochemistry and real-time quantitative polymerase chain reaction (RT-Q-PCR)by SYBR Green. And the standardized cycle of threshold(SCt)was regarded as the indicators of the mRNA expression. Results HS and GBM thickness were significantly higher in DM rats than those in NDM rats,especially in DM rats of the courses of 20 and 24 weeks(P < 0.01). IHC scores of VEGF and VEGFR2 were higher in total DM rats and were positively correlated with each other. There were positive correlations between HS with VEGF and VEGFR2 in total DM rats(P < 0.05). SCts of VEGF and VEGFR2 were significantly higher and were positively correlated with each other in total DM rats(P < 0.01). SCt of VEGF and GBM thickness showed positive correlation in total DM rats. SCt of VEGF was highest in the course of 12w DM rats. SCt of VEGFR2 gradually decreased following by the diabetic course,and was lowest in the course of 24w. There were no significantly differences in IHC scores and SCt of mTOR between DM and NDM rats. But the IHC scores of mTOR,VEGF and VEGFR2 were positively correlated with each other and with HS in total DM rats(P < 0.05). Conclusion HS and GBM thickness were higher in diabetic rats,especially in the course of 24w,which could reflect the injury of nephropathy. The protein and mRNA of VEGF and VEGFR2 were high expressed in kidney of DM rats and increased with the increasing of diabetic course. The mRNA expression of VEGF was positively correlated with GBM thickness of in diabetic nephropathy(DN). The protein expressions of mTOR,VEGF and VEGFR2 might have synergistic effects in DN of DM rats. But the results could not exclude the influences of different courses,sample size and experimental conditions.

Freshwater crabs (Sinopotamon denticulatum) were examined for metacercariae. Cats and dogs were also examined for Paragonimus infection. Questionnairing was carried out on health knowledge and behaviors among local residents in a village of Baokang County, Hubei Province. Results showed that the infection rate of Paragonimus skrjabini metacercariae in Sinopotamon denticulatum was 20.5% (46/214), with 15.6% (20/128) in a mining area and 30.2%(26/86) for the non-mining area respectively ( ?2=6.5, P0.05). Questionnairing showed that dogs and cats were with the habit of foraging and defecating at streams and children had the habits of eating raw or under-cooked crabs. The natural and ecological environments are in favor of the life cycle of P. skrjabini.

Objective: This study used cardiac magnetic resonance imaging (MRI) to compare the characteristics of left ventricular remodeling in patients with primary aldosteronism (PA) with those of patients with essential hypertension (EH) and healthy controls (HCs). Materials and Methods: This prospective study enrolled 35 patients with PA, in addition to 35 age- and sex-matched patients with EH, and 35 age- and sex-matched HCs, all of whom underwent comprehensive clinical and cardiac MRI examinations. The analysis of variance was used to detect the differences in the characteristics of left ventricular remodeling among the three groups. Univariable and multivariable linear regression analyses were used to determine the relationships between left ventricular remodeling and the physiological variables. Results: The left ventricular end-diastolic volume index (EDVi) (mean ± standard deviation [SD]: 85.1 ± 13.0 mL/m2 for PA, 75.9 ± 14.3 mL/m2 for EH, and 77.3 ± 12.8 mL/m2 for HC; p = 0.010), left ventricular end-systolic volume index (ESVi) (mean ± SD: 35.2 ± 9.8 mL/m2 for PA, 30.7 ± 8.1 mL/m2 for EH, and 29.5 ± 7.0 mL/m2 for HC; p = 0.013), left ventricular mass index (mean ± SD: 65.8 ± 16.5 g/m2 for PA, 56.9 ± 12.1 g/m2 for EH, and 44.1 ± 8.9 g/m2 for HC; p < 0.001), and native T1 (mean ± SD: 1224 ± 39 ms for PA, 1201 ± 47 ms for EH, and 1200 ± 44 ms for HC; p = 0.041) values were higher in the PA group compared to the EH and HC groups. Multivariable linear regression demonstrated that log (plasma aldosteroneto-renin ratio) was independently correlated with EDVi and ESVi. Plasma aldosterone was independently correlated with native T1. Conclusion Patients with PA showed a greater degree of ventricular hypertrophy and enlargement, as well as myocardial fibrosis, compared to those with EH. Cardiac MRI T1 mapping can detect left ventricular myocardial fibrosis in patients with PA.

Objective: This study used cardiac magnetic resonance imaging (MRI) to compare the characteristics of left ventricular remodeling in patients with primary aldosteronism (PA) with those of patients with essential hypertension (EH) and healthy controls (HCs). Materials and Methods: This prospective study enrolled 35 patients with PA, in addition to 35 age- and sex-matched patients with EH, and 35 age- and sex-matched HCs, all of whom underwent comprehensive clinical and cardiac MRI examinations. The analysis of variance was used to detect the differences in the characteristics of left ventricular remodeling among the three groups. Univariable and multivariable linear regression analyses were used to determine the relationships between left ventricular remodeling and the physiological variables. Results: The left ventricular end-diastolic volume index (EDVi) (mean ± standard deviation [SD]: 85.1 ± 13.0 mL/m2 for PA, 75.9 ± 14.3 mL/m2 for EH, and 77.3 ± 12.8 mL/m2 for HC; p = 0.010), left ventricular end-systolic volume index (ESVi) (mean ± SD: 35.2 ± 9.8 mL/m2 for PA, 30.7 ± 8.1 mL/m2 for EH, and 29.5 ± 7.0 mL/m2 for HC; p = 0.013), left ventricular mass index (mean ± SD: 65.8 ± 16.5 g/m2 for PA, 56.9 ± 12.1 g/m2 for EH, and 44.1 ± 8.9 g/m2 for HC; p < 0.001), and native T1 (mean ± SD: 1224 ± 39 ms for PA, 1201 ± 47 ms for EH, and 1200 ± 44 ms for HC; p = 0.041) values were higher in the PA group compared to the EH and HC groups. Multivariable linear regression demonstrated that log (plasma aldosteroneto-renin ratio) was independently correlated with EDVi and ESVi. Plasma aldosterone was independently correlated with native T1. Conclusion Patients with PA showed a greater degree of ventricular hypertrophy and enlargement, as well as myocardial fibrosis, compared to those with EH. Cardiac MRI T1 mapping can detect left ventricular myocardial fibrosis in patients with PA.

Objective: To clinically validate the feasibility and accuracy of cine images acquired through the multitasking method, with no electrocardiogram gating and free-breathing, in measuring left ventricular (LV) function indices by comparing them with those acquired through the balanced steady-state free precession (bSSFP) method, with multiple breath-holds and electrocardiogram gating. Materials and Methods: Forty-three healthy volunteers (female:male, 30:13; mean age, 23.1 ± 2.3 years) and 36 patients requiring an assessment of LV function for various clinical indications (female:male, 22:14; 57.8 ± 11.3 years) were enrolled in this prospective study. Each participant underwent cardiac magnetic resonance imaging (MRI) using the multiple breath-hold bSSFP method and free-breathing multitasking method. LV function parameters were measured for both MRI methods. Image quality was assessed through subjective image quality scores (1 to 5) and calculation of the contrast-to-noise ratio (CNR) between the myocardium and blood pool. Differences between the two MRI methods were analyzed using the Bland–Altman plot, paired t-test, or Wilcoxon signed-rank test, as appropriate. Results: LV ejection fraction (LVEF) was not significantly different between the two MRI methods (P = 0.222 in healthy volunteers and P = 0.343 in patients). LV end-diastolic mass was slightly overestimated with multitasking in both healthy volunteers (multitasking vs. bSSFP, 60.5 ± 10.7 g vs. 58.0 ± 10.4 g, respectively; P < 0.001) and patients (69.4 ± 18.1 g vs. 66.8 ± 18.0 g, respectively; P = 0.003). Acceptable and comparable image quality was achieved for both MRI methods (multitasking vs. bSSFP, 4.5 ± 0.7 vs. 4.6 ± 0.6, respectively; P = 0.203). The CNR between the myocardium and blood pool showed no significant differences between the two MRI methods (18.89 ± 6.65 vs. 18.19 ± 5.83, respectively; P = 0.480). Conclusion Multitasking-derived cine images obtained without electrocardiogram gating and breath-holding achieved similar image quality and accurate quantification of LVEF in healthy volunteers and patients.