Objective To establish a practical,stable and high purity endothelial progenitor cells culture meth- od in vitro.Methods Human umbilical cord blood mononuclear cells were isolated by Ficoll density-gradient cen- trifugation,then plated on dishes coated with human fibronectin.After 48 hours,the nonaderent cells were collect- ed and replated onto fibronectin-coated dishes.After 7 days of culture,the cells were identified with the techniques of immunohistochemistry,immunofluorescence and flow cytometer.Results The cultured cells were small and spindle or polygonal in shape.Large numbers of typical endothelial progenitor cell colony-forming units were found,vWF and Flk-1 proteins expression were identified in more than 95% of the attached cells with 98% of them showing positive Dil-ac-LDL and FITC-UEA-1.According to the results from fluorescence-activated cell sorting (FACS),7.0%?1.8% of cells were recognized as CD_(133)~+.Conclusion Differential attachment technique is a practical and stable method for obtaining highly purified endothelial progenitor cells.

Objective To explore the effects of diabetes mellitus on the left ventricular geometry in essential hypertension(EH). Methods Patients with hypertension admitted between 2000 to 2007 were retrospectively analysed.Those meeting the study criteria were divided into EH group(n=1660) and EH with diabetes mellitus group(n=953).The general data,biochemical parameters and echocardiographic findings were analysed. Results Compared with EH group,the age,course of hypertension,body mass index,systolic blood pressure,fasting blood glucose and triglyceride were increased(P

Objective: To evaluate the efficacy of Tuina (Chinese therapeutic massage) manipulation plus horse-riding squat exercise in treating knee osteoarthritis (KOA) and optimize the combining protocol. Methods: Based on a 2×2 factorial design, 120 eligible KOA patients were randomized into a manipulation group (group A1B2), a manipulation plus horse-riding squat group (group A1B1), a sitting knee-adjustment group (group A2B2 group), and a sitting knee-adjustment plus horse-riding squat group (group A2B1), with 30 cases in each group. The intervention was conducted three times a week, lasting for four weeks. The Western Ontario and McMaster Universities osteoarthritis index (WOMAC) was taken as the major measure for efficacy evaluation (including three component scores, pain, stiffness, and daily function, and total score). Results: The three component scores (pain, stiffness, and daily function) and the total score of WOMAC showed significant differences after the intervention in the four groups (P<0.05). There were significant inter-group differences in the WOMAC stiffness score amongst the four groups after the intervention (P<0.05). In group A1B1, the step length, stride, walking speed, and knee joint flexion angle changed significantly after treatment (P<0.05). After the intervention, the step length changed significantly in group A1B2 (P<0.05), and the walking speed changed significantly in group A2B1 (P<0.05). There were no significant differences in the step length, stride, walking speed, or knee joint flexion angle among the four groups (P>0.05). The extensor peak torque at 180 °/s changed significantly in group A1B2 after treatment (P<0.05). Neither the intra-group nor the inter-group comparisons of the four groups revealed significant differences in the other isokinetic muscle strength parameters (P>0.05). The main effect of manipulation showed significant in affecting the WOMAC pain and total scores (P<0.05). The main effect of horse-riding squat exercise showed significant in affecting the WOMAC pain and stiffness scores (P<0.05). Conclusion: The four treatment protocols all can improve the symptoms of KOA, for instance, relieving pain and stiffness, and enhancing daily function. Group A2B1 produces the most eminent effect in relieving joint stiffness. The main effects of both manipulation and horse-riding squat exercise are significant in reducing pain. Besides, the main effect of horse-riding squat exercise is significant in relieving joint stiffness.

AIM: To investigate the immune stimulation capacity of B7-H1 blockade on immature dendritic cells (DCs) in vitro. METHODS: The human monocyte-derived dendritic cells were induced in the presence of cytokine GM-CSF and IL-4. The expression of B7-H1 was detected by FCM. On blockade of B7-H1, the maturation and endocytic activity, T cells stimulatory proliferation capacity, IL-12 production, T cell differentiation effect of DCs were detected by FCM, MTT assay, ELISA and ELISPOT, respectively. RESULTS: The expression of B7-H1 was increased with the induction of DCs. On day 7, the positive expression was 54.12%, and the TNF-? induced mature DCs had the positive expression rate of 83.64%. The blockade of B7-H1 on immature DCs had sharply increased their T cells stimulatory proliferation capacity and IL-12 production, and efficiently induced the development of Th1/Tc1 cells, but had no effect on their maturation and endocytic activity. CONCLUSION: The blockade of B7-H1 on immature DCs increases its immune stimulation activity. It is valuable to investigate the antitumor immune responses of DCs vaccine with B7-H1 blockade.

Objective To discuss how to protect the intracranial vertebral artery and posterior inferior cerebellar artery by observing and measuring the intracranial vertebral artery in the surgery adopt far lateral approach. Methods Mimicking far lateral approach, 20 adult cadaveric heads connected to neck fixed with 10% formalin were dissected. Intracranial segment of the vertebral arteries and their main branches were exposed and measured under operating microscope. Results The intracranial vertebral artery joined with the contralateral one into the basilar artery after traveling through the atlanto-occipital sulcus. The relationship between the vertebral artery and the hypoglossal nerve is close. Thirty sides (75%) of the vertebral arteries traveled to pons medulla sulcus in front of the hypoglossal nerve roots and 2 sides (5%) behind the hypoglossal nerve roots, while 8 sides (20%) traveled among the hypoglossal nerve roots; 70% of the vertebral arteries were contacted to the hypoglossal nerve roots, 30% of which compressed the hypoglossal nerve. The main branches of intracranial segment of the vertebral arteries were the posterior inferior cerebellar arteries, the anterior spinal arteries, the posterior meningeal arteries,and some perforating arteries. Posterior inferior cerebellar arteries all originated from the intracranial vertebral artery were the largest vertebral artery's branches; their trip was mostly loop-shaped and they had close relationship with Ⅸ, Ⅹ, Ⅺ cranial nerves. The starting points of the posterior inferior cerebellar arteries were different, even in the same specimen, but most of them originated from the upper 1/3intracranial vertebral artery. No anterior inferior cerebellar artery was noted originated from the vertebral artery in our specimen. Anterior spinal arteries originated from the vertebral arteries joined with the branches of the bilateral vertebral arteries and traveled down through the tortuous anterior median fissure to supply the spinal cord. Conclusion Being familiar with the characteristics and anatomic vertebral arteries variations of the intracranial vertebral artery and its branches can contribute to identify and protect the intracranial segment of the vertebral artery and its main branches in the surgery adopt far-lateral approach.

Objective:To investigate the relationship between body mass index (BMI) and response time of cardioinhibitory type vasovagal syncope (VVS-CI) in children.Methods:The clinical data of 56 children with syncope or pre-syncope were retrospectively analyzed and they visited specialist clinic for syncope and were diagnosed as VVS-CI in the Second Xiangya Hospital, Central South University from December 2012 to September 2019.Based on height and weight, BMI was calculated, and divided into low BMI group (35 cases) and normal BMI group (21 cases). Between the 2 groups, baseline heart rate, head-up tilt test (HUTT) positive response heart rate, baseline head-up tilt test (BHUT) positive response time, and sublingual nitroglycerin-provocated HUTT (SNHUT) positive response time were compared.The correlation between BMI and positive response time was analyzed.SPSS 22.0 software was applied for statistical analysis.Results:There were no significant differences in age, sex, duration of disease and number of syncope between the 2 groups (all P>0.05). No significant differences were found in baseline heart rate and positive response heart rate between the 2 groups [(78.5±15.3) times/min vs.(72.8±8.7) times/min, t=1.223, P=0.230; (44.0±13.9) times/min vs.(47.0±10.0) times/min, t=-0.664, P=0.511]. Compared with normal BMI group, BHUT positive patients/SNHUT positive patients were higher in low BMI group (27/8 cases vs.9/12 cases, χ2=4.839, P=0.027), and the positive response time of BHUT was shorter [(13.1±4.6) min vs.(23.7±9.5) min, t=-2.691, P=0.023]. There were no significant differences in SNHUT positive response time between the 2 groups ( P>0.05). Low BMI was correlated with BHUT positive response time ( r=0.750, P=0.005). Normal BMI was not associated with BHUT positive response time ( r=0.316, P=0.217). There was no correlation between low BMI and normal BMI and SNHUT positive response time ( r=0.177, P=0.431; r=0.021, P=0.940). Conclusions:Low BMI is positively correlated with BHUT positive response time of children with VVS-CI.The time it takes for syncope occurrence was shorter in children with low BMI than that in normal BMI.

Objective:To investigate the difference of heart rate variability in cardioinhibitory type vasovagal syncope(VVS-CI) children with different body mass index(BMI).Methods:Clinical data of thirty-four children with syncope or pre-syncope were retrospectively analyzed, who visited specialist clinic for syncope and diagnosed as VVS-CI at the Second Xiangya Hospital of Central South University from January 2012 to December 2019.BMI was calculated based on height and weight, and divided into lean group(BMI≤18.4 kg/m 2, n=19) and normal group(BMI 18.5-23.9 kg/m 2, n=15). Heart rate variability(HRV) of 24 h dynamic electrocardiogram was analyzed using linear analysis method.Time domain index included SDNN, SDANN, rMSSD and pNN50.Frequency domain index included total power(TP), ultra low frequency power(ULF), very low frequency power(VLF), low frequency power(LF), high frequency power(HF) and LF/HF. Results:There was no significant difference in SDNN, SDANN and rMSSD between lean and normal group( P>0.05), but pNN50 increased in lean group( P<0.05). No significant differences were found in TP, ULF, LF, HF and LF/HF between two groups( P>0.05), while VLF was lower in lean group than that in normal group( P<0.05). There was no statistical difference in time domain index and frequency domain index between different gender between lean and normal group( P>0.05). SDNN, SDANN and LF were higher in<12 years old than those in≥12 years old in lean group( P<0.05). There was no statistical difference in rMSSD, pNN50, TP, ULF, VLF, HF and LF/HF( P>0.05). ULF increased and LF decreased in<12 years old compared to ≥12 years old in normal group( P<0.05). No statistical differences were found in SDNN, SDANN, rMSSD, pNN50, TP, VLF, HF and LF/HF( P>0.05). Conclusion:The autonomic nervous regulation function of VVS-CI children with low BMI and normal BMI is different, resulting in HRV difference.There were also differences in HRV between<12 years old and ≥12 years old with the same BMI.

Objective To investigate the effect of insulin-like growth factor- Ⅰ ( IGF- Ⅰ ) on the proliferation and osteogenesis of human periodontal ligament stem cells ( hPDLC ) under three-dimensional (3D) culture system. Methods Human periodontal cells were isolated from the ligament of surgically extracted human teeth, and through the limiting dilution assay, got mono-clone of the cell, hPDLCs were isolated from MesenPRO RS medium. Rotary cell culture system (RCCS) was enrolled to set 3D environment. Control group and experiment groups were assigned according to the concentration of IGF- Ⅰ .There were 5 level of experiment groups (0.1,1,10,50,100 μg/L). Proliferation was tested with methyl thiazolyl tetrazolium ( MTT), and alkine phosphatase (ALP) level was assayed by spectrophotometer to analyze the osteogenesis of hPDLCs. Gene expression of ostetocalcin(OCN)and type Ⅰ collagen (Col Ⅰ )were assayed by reverse transcriptase polymerase chain reaction(RT-PCR). Results In 3D culture system,the effect of IGF- Ⅰ on cell proliferation was significantly different between control group and experiment groups( P ＜ 0.05 ), and there showed significant differences between the group of 0.1 μg/L ( 0.219 ±0.021 ) IGF- Ⅰ and the groups of 50, 100 μg/L(0.287 ±0.011,0.293 ±0.012). However, there showed no significant differences among other groups. Significant differences of ALP activity were observed between the control group and experiment groups, and between the groups of 1, 10 μg/L(0.304 ±0.020, 0.310 ±0.013) and that of 50, 100 μg/L (0.347 ±0.011, 0.344 ±0.010) (P ＜0.05). While no significantdifferences were detected between the group of 1 μg/L and that of 10 μg/L, nor between the group of 50 μg/L and that of 100 μg/L. Expressions of Col Ⅰ and OCN in mRNA and protein level both showed dose-dependent increase. Conclusions In 3D culture system, in the scale of 0.1-100 μg/L, the effect of IGF-Ⅰ on the proliferation of hPDLCs increased dose-dependently. 100 μg/L IGF- Ⅰ promotes osteogenesis of the cells significantly.

OBJECTIVETo investigate the influential factors of total blood loss during unilateral artificial total hip arthroplasty (THA).

METHODSFrom January 2014 to July 2016, 131 patients undergwent primary unilateral total hip arthroplasty, including 55 males and 76 females, ranging in age from 40 to 89 years old, with an average of 64.5 years old. The data of patients with unilateral total hip arthroplasty were collected, and the data of hemodynamics, coagulation function, hemoglobin and hematocrit values were recorded according to the observation data. The blood loss of the patients was calculated by Gross equation, and the data were analyzed by a statistical software.

RESULTSThe total blood loss of the patients was significantly affected by the factors of pathogenesis and deep venous thrombosis of the lower limbs. The results were statistically significant(<0.05). The gender, age, body mass index, prosthesis and anesthesia had little effects on the blood loss volume after replacement.

CONCLUSIONSThe data in this study indicate that the volume of actual blood loss can be controlled by adjusting non-surgical factors such as gender, age, body mass index, prosthetic material, and risk factors, and surgical factors(postoperative complications, anesthesia).

Objective: To evaluate the effectiveness of therapeutic massage (tuina) for treating knee osteoarthritis (KOA). Methods: Six English and Chinese databases, including Chinese National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP), China Biology Medicine Disc (CBM), Cochrane Library and PubMed databases, were independently searched to identify appropriate randomized controlled trials (RCTs) studying therapeutic massage for KOA compared to oral non-steroidal anti-inflammatory drugs (NSAIDs) alone. The main outcome measures were total effectiveness and the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) score. Results: A total of 8 RCTs were included and they were of average quality. The results showed that therapeutic massage was more effective than NSAIDs comparing total effectiveness [risk ratio (RR)=1.14, 95％ confidence interval (CI) (1.07, 1.21), P<0.0001]; compared with NSAIDs, therapeutic massage produced more significant improvements in pain [mean difference (MD)=-2.06, 95％CI (-2.75, -1.36), P<0.00001], stiffness intensity [MD=-0.90, 95％CI (-1.05, -0.75), P<0.00001] and joint function [MD=-12.48, 95％CI (-13.91, -11.05), P<0.00001]. Conclusion: Therapeutic massage was more effective than oral NSAIDs in treating KOA. In relieving pain and stiffness and improving the function of knee joint, therapeutic massage was superior to NSAIDs.