BACKGROUND: Previous studies have confirmed that rabbit bone marrow mesenchymal stem cells (BMSCs) can differentiate into osteoblasts under osteogenic induction in vitro, stably express the specific phenotype of osteoblasts and have osteogenic ability. Calf cortical bone scaffold with partial cancellous bone has good biocompatibility and degradability, which can be used as a carrier material of bone marrow mesenchymal stem cells. OBJECTIVE: To combine rabbit BMSCs with calf bone composite according to the basic principles of bone tissue engineering and to observe the osteogenesis in the New Zealand white rabbits after implantation of BMSCs/calf bone composite into the ilium, thereby providing a direct evidence for preliminary clinical application of tissue-engineered bone products.METHODS: BMSCs/calf cortical bone scaffold with partial cancellous bone (tissue-engineered bone group), simple calf heterogeneous bone (heterogeneous bone group) or autologous iliac bone (autologous iliac bone group) was randomly implanted into the rabbit ilium. The changes of implant surface and tissue reactions around the implant were observed.X-ray examination was performed to observe osteogenic changes at 4, 8, 12, 24 weeks after implantation. Immunohistochemistry staining was used to observe the expression of bone morphogenetic protein 2.RESULTS AND CONCLUSION: After heterogeneous bone implantation, the wound healed well, and there were no systemic or local inflammation and toxicity reactions in all groups. The X-ray results showed that at postoperative 24 weeks, the implant was basically fused with the host bone in the tissue-engineered bone group, but the fusion was unsatisfactory in the heterogeneous bone group. The process of ossifications from cartilages was observed in all groups by hematoxylin-eosin staining, and bone morphogenetic protein 2 was positive for immunohistochemical staining. Findings from in vivo experiments indicate that rabbit BMSCs seeded onto the calf cortical bone scaffold with partial cancellous bone could construct tissue-engineered bone by osteoinductation in vitro in the rabbits.

Objective To investigate the clinical significance of tumor related gene gankyrin expression in pancreatic cancer.Methods The immunohistochemical staining was performed to study the gankyrin expression in 62 pairs of pancreatic cancer and matched non-tumor tissues.The correlations ofgankyrin expression with pathological parameters and the influences on the prognosis were analyzed.Results Ggankyrin positivity in pancreatic cancer and matched non tumor tissue was 2.5％ (45/62)and 19.4％ (12/62),respectively.The positivity of gankyrin was significantly higher in pancreatic cancer compared with matched non-tumor tissues,and the differences were statistical significant (P ＜ 0.01).Gankyrin expression in pancreatic cancer tissues were significantly correlated with TNM stage,differentiation and metastasis (P ＜ 0.05),but not with age,gender or tumor location (P ＞ 0.05).The 3-year survival rates of patients with positive and negative gankyrin expression were 17.8 ％ (8/45)and 41.2％ (7/17),respectively.Patients with positive gankyrin expression had a significantly shorter survival period compared with these with negative expression in pancreatic cancer tissues (P =0.034).Conclusions Gankyrin was overexpressed in pancreatic cancer,which was correlated with poor prognosis.

Objective To isolate and purify Thy 1 low Lin Sca 1 + bone marrow stem subset cells by magnetic activated cell sorting(MACS). Methods Thy 1 low Lin Sca 1 + cells from mouse bone marrow were collected through three processes by MACS.After Lin + cells were removed,Thy 1 low Lin Sca 1 + bone marrow stem cell subsets were harvasted.The purity of the cells was analysed by FACS and the reclaimation rate was counted. Results The purity and reclaimation rate of Thy 1 low Lin Sca 1 + cells were 72.36% and 82.43% respectively, which equaled to the level of isolation and purification of CD34 + cells by MACS. Conclusions It is effective to isolate and purify Thy 1 low Lin Sca 1 + bone marrow stem cell subsets by MACS, and the purity and reclaimation rate of the cells are high.

This study examined the current changes of human ether-a-go-go-related gene (hERG) mutation derived from a LQT2 Chinese family with a highly penetrating phenotype. Mutation was identified and site-directed mutagenesis was performed to induce the mutation in wild-type (WT) hERG. WT hERG and mutated V535M were cloned and transiently expressed in HEK293 cells. At the 48th and 72nd h after transfection, membrane currents were recorded using whole cell patch-clamp procedures. An A>G transition at 1605 resulting in replacement of V535M was identified. Compared to WT, V535M mutation significantly decreased tail currents of hERG. At test potential of -40 mV after depolarizing at +50 mV, tail current densities were 83.35±7.06 pA/pF in WT and 50.38±7.74 pA/pF in V535M respectively (n=20, P<0.01). Gating kinetics of hERG revealed that V (1/2) of steady-state inactivation shifted to negative potential in the mutant (V (1/2,V535M): -61.81±1.7 mV vs. V (1/2, WT): -43.1±0.71 mV). The time constant of recovery from inactivation was markedly prolonged in the mutant compared to WT among test potentials. V535M hERG mutation demonstrated markedly decreased tail current densities, which suggests that V535M is a new loss-of-function mutation of hERG channel responsible for LQT2.

Objective To analyze the immune function and shoulder function of sentinel lymph node biopsy combined with mastectomy in patients with early breast cancer.Methods A total of 80 patients with breast fibroadenoma were selected ,and they were randomly divided into surround -mammary areola incision group ( n=40 ) and conven-tional radial incision group(n=40) according to the digital table.The bleeding volume,operation time,hospital stay, satisfaction and curative effect were compared between the two groups .Results The markedly effective rate of surround-mammary areola incision group was 87.50％,which was significantly higher than that of the radial incision group(62.50％),and the difference was statistically significant (χ2 =10.400,P=0.001).The total effective rate was 100.00％in the surround -mammary areola incision group ,which was significantly higher than that in the radical incision group(χ2 =6.500,P=0.010).The time of operation,the amount of bleeding and the length of hospital stay in the surround-mammary areola incision group were (56.7 ±3.2) min,(62.1 ±1.8) mL and(8.3 ±0.8) d, respectively,which were lower than those in the radial incision group [(69.8 ±4.3)min,(71.1 ±2.4)min,(11.2 ± 1.3)d],there were statistically significant differences between the two groups (t=15.262,P=0.000;t=18.735, P=0.000;t=11.864,P=0.000).The incidence rates of breast milk symmetry,scar acceptance and adverse reaction in surround-mammary areola incision group were 87.50％,87.50％ and 2.50％,respectively,which were significantly better than those in the radial incision group(67.50％,65.00％ and 15.00％),there were statistically significant differences between the two groups (χ2 =5.032,P=0.024;χ2 =7.862,P=0.005;χ2 =3.923,P=0.047).The total satisfaction rate was 87.50％in the surround-mammary areola incision group ,which was higher than 62.50％in the radial incision group (χ2 =7.222,P=0.007).Conclusion The use of surround -mammary areola incision in the treatment of patients with breast fibroadenoma has obvious effect ,can reduce the degree of breast injury ,improve the physical appearance of the breast ,with high satisfaction .

This study examined the current changes of human ether-a-go-go-related gene (hERG) mutation derived from a LQT2 Chinese family with a highly penetrating phenotype.Mutation was identified and site-directed mutagenesis was performed to induce the mutation in wild-type (WT) hERG.WT　hERG and mutated V535M were cloned and transiently expressed in HEK293 cells.At the 48th and 72nd h after transfection,membrane currents were recorded using whole cell patch-clamp procedures.An A＞G transition at 1605 resulting in replacement of V535M was identified.Compared to WT,V535M mutation significantly decreased tail currents of hERG.At test potential of-40 mV after depolarizing at +50 mV,tail current densities were 83.35±7.06 pA/pF in WT and 50.38±7.74 pA/pF in V535M respectively (n=20,P＜0.01).Gating kinetics of hERG revealed that V1/2 of steady-state inactivation shifted to negative potential in the mutant (V1/2,V535M:-61.81±1.7 mV vs.V1/2,WT:-43.1±0.71mV).The time constant of recovery from inactivation was markedly prolonged in the mutant compared to WT among test potentials.V535M hERG mutation demonstrated markedly decreased tail current densities,which suggests that V535M is a new loss-of-function mutation of bERG channel responsible for LQT2.

Objective:To explore the differences in clinical and imaging features between pancreatic adenosquamous carcinoma (PASC) and pancreatic ductal adenocarcinoma (PDAC).Methods:The clinical data, imaging and pathological data of 171 patients pathologically diagnosed with PASC after surgical resection (PASC group) (from February 2011 to October 2020, 148 patients from the First Affiliated Hospital of Naval Medical University and 23 patients from the Second Affiliated Hospital of Zhejiang University School of Medicine) and 100 patients pathologically diagnosed with PDAC after surgical resection (PDAC group) (from January to June, 2018, at the First Affiliated Hospital of Naval Medical University) were retrospectively analyzed. Computed tomography and magnetic resonance imaging features were analyzed by two associate chief physician of department of radiology. Independent sample t test, rank sum test, chi-square test or Fisher exact probability test were used for statistical analysis. Multivariate logistic regression analysis was used to analyze independent predictors of PASC. Results:The longest diameter of tumor of PASC group was larger than that of PDAC group (35.0 mm (28.0 mm to 45.0 mm) vs. 29.5 mm (23.0 mm to 36.0 mm)), and the rates of cystic necrosis, ring-enhancement, normal distal main pancreatic duct and normal pancreatic parenchyma of PASC group were higher than those of PDAC group (62.0%, 106/171 vs. 12.0%, 12/100; 66.1%, 113/171 vs. 25.0%, 25/100; 52.0%, 89/171 vs. 12.0%, 12/100; 70.2%, 120/171 vs. 29.0%, 29/100, respectively); and the differences were statistically significant ( Z=-4.001, χ2=72.183, 42.612, 43.284 and 43.221, all P<0.01). The results of multivariate logistic regression analysis showed that the cystic necrosis, ring-enhancement, normal distal main pancreatic duct and normal pancreatic parenchyma were indenpendent predictors of PASC (odds ratio=10.083, 2.361, 3.086 and 2.632, 95% confidence interval 8.736 to 11.639, 2.096 to 2.660, 2.605 to 3.656 and 2.267 to 3.057, all P<0.01); and the sensitivity for PASC diagnosis was 62.0%, 66.1%, 51.7% and 70.3%, respectively; the specificity was 88.0%, 75.0%, 88.0% and 71.0%, respectively; the positive predictive value was 89.3%, 81.9%, 88.1% and 80.5%, respectively. Conclusions:PASC and PDAC have similar clinical features. The imaging features of cystic necrosis, ring-enhancement, normal distal main pancreatic duct and normal pancreatic parenchyma are independent predictive factors of PASC.

Objective To investigate the value of real-time elastography (RTE) in evaluating the characteristics and elasticity index (E-index) changes at different echo level of uterine fibroids before and after radiofrequency ablation (RFA).Methods A total of 43 patients (with 55 lesions) accepted RFA under ultrasound guidance in the Fourth Affiliated Hospital of Harbin Medical University from September 2013 to January 2015.All lesions were confirmed to have no perfusion through ultrasonography 1 h after RFA.The diameter of lesions ranging from 1.0 cm to 3.0 cm (with an average of 1.9± 1.2 cm).Preoperative lesions were classified into 3 groups for comparison based on ultrasonic echo type,group A:hypo echo (n=30);group B:hyper echo (n=12);and group C:mixed echo (n=12).Measured elasticity index E-index and internal uniformity (△ E) were analyzed.Comparisons of E value and△ E value were performed with repeated measures.RTE images features using x2 test of Fisher inspection comparison at the same time differences between groups.Results (1) Before RFA,RTE images showed blue alternating with green with green being dominated accounting for 70％ (21/30).50％ of RTE images in group B were completely covered in blue (6/12),while group C exhibited 53.9％ blue alternating with green with blue being dominated (7/13).There were statistically significant characteristics of RTE images among 3 groups (P ＜ 0.01).1 h after RFA,74.5％ lesions exhibited with blue being dominated (41/55).However,83.6％ (41/55) exhibited with blue 3 months after RFA.The difference in characteristics of RTE among 3 groups displayed no statistical significance (P ＞ 0.05).(2) The comparison of E-index revealed that before RFA,the E value was the lowest in the group A and the highest in the group B,while the group C was between them.There was significant difference between group A and B (F=19.25,P ＜ 0.01).E-index elevated in all the three groups and significant differences were found 1 h,3 month after RFA in comparison with that before RFA in the group A (F=386.75,294.68,both P ＜ 0.01).In addition,significant differences were also found 1 h,3 month after RFA in comparison with the treatment in the group B (F=29.98,45.88,both P ＜ 0.01).As for the group C,there were significant differences in E-index 1 h,3 month after RFA compared with before RFA (F=120.29,139.64,both P ＜ 0.01).3 months after RFA,E-index further elevated in the three groups and no significant inter-group difference was found at the same time points (P ＞ 0.05).(3)Before RFA,the△ E value was lower in group A and group B,while higher in the group C,group C presenting significant difference from group A and B (F=484.68,344.15,both P ＜ 0.01).At 1 h after RFA,△ E value was higher in group A and group C,while lower in group B,with group B showing significant difference from the low and mixed echo groups (F=53.58,94.79,both P ＜ 0.01).Significant difference was observed 1 h after surgery compared to before RFA in group A (F=154.35,P ＜ 0.01).Significant difference was also found 1 h after RFA compared to 3 month after RFA in group A (F=266.85,P ＜ 0.01).As for group C,there were also significant differences in△ E-index before treatment and 1 h after RFA compared with 3 month after RFA (F=103.24,76.53,both P ＜ 0.01).At 3 months after RFA,△ E-index decreased in all three groups.No significant inter-group difference was found at the same time points (all P ＞ 0.05).Conclusions RTE characteristics and the elasticity index are distinctly different among uterine fibroid at various echo intensities.RTE contributes to judging changes in hardness before and after RFA.The application of RTE for quantitative comparison of lesion hardness and uniformity can be served as the foundation for evaluating therapeutic effects of RFA.

Objective:To document any effect of environmental enrichment on nerve regeneration in a mouse model of sciatic nerve compression and explore its mechanism.Methods:A crushed sciatic nerve model was successfully established in 22 C57BL/6 mice, and they were then randomly divided into an intervention group and a control group. The mice of the intervention group were raised in a cage with an enriched environment, while those of the control group were kept in a standard cage. Two weeks later, both groups′ gait was analyzed and the compound muscle action potential (CMAP) of the sciatic nerve was measured. The proportion of myelinated sciatic nerve fibers was examined using toluidine blue staining, and the expression of myelin basic protein (MBP), growth associated protein-43 (GAP43) and p75 neurotrophin receptor (p75 NTR) was measured using immunofluorescence intensity. Results:①The latency of the CMAP [(1.05±0.04)ms] was significantly shortened in the intervention group compared with the control group and the amplitude was significantly higher. ②Gait analysis showed a significant increase in the average contact intensity, stride length and stride rate of the intervention group compared with the control group. However, the step axis angle of the intervention group was significantly smaller than in the control group on average. ③The stained nerve fibers in the intervention group were orderly and dense, and the average number of myelinated fibers was significantly greater than in the control group. ④Quantitative analysis of the immunofluorescence showed that the levels of MBP, GAP43 and p75 NTR in the sciatic nerves of the intervention group were, on average, significantly higher than in the control group. Conclusion:An enriched environmental can promote the regeneration and functional recovery of crushed sciatic nerves by promoting the proliferation and myelination of Schwann cells.

OBJECTIVETo evaluate the application of inferior vena cava filter (IVCF) in prevention of peri-operative pulmonary embolism (PE) in lower limb bone fracture patients with deep venous thrombosis (DVT).

METHODSFrom January 2003 to December 2012, 2 248 cases of lower limb and pelvic fractures with DVT were retrospectively analyzed. Before the procedure of IVCF implantation began, January 2003 to December 2007, there were 1 052 cases of acute trauma patients with DVT were classified as the group of early none-IVCF. The IVCF implantation was began since January 2008. From that time to December 2012, 712 cases of bone fractures with DVT received filter implantation, which were classified as IVCF group. The other 484 patients who had not undergone filter deployment were divided as group of late none-IVCF. The baseline conditions of the three groups were significantly different in addition to the ages between group of early none-IVCF and IVCF group. The incidences of PE and mortality of PE in each group were recorded and analyzed by χ(2) test.

RESULTSThere were totally 31 cases of symptomatic PE, among which 12 cases died. Totally 712 filters were deployed successfully without any major complications. The incidences of symptomatic PE were 0.14% (1/712), 2.19% (23/1 052) and 1.45% (7/484) in IVCF group, group of early none-IVCF and group of late none-IVCF, respectively. The mortality of PE were 0 (0/712), 0.86% (9/1 052) and 0.62% (3/484) in these groups. The incidence of symptomatic PE in IVCF group was significantly different from that in the group of early and late none-IVCF (χ(2) = 11.762, P = 0.001; χ(2) = 7.395, P = 0.007, respectively). The mortality of IVCF group was also significantly lower compared with the other two groups (χ(2) = 6.122, P = 0.013; χ(2) = 4.424, P = 0.035, respectively).

CONCLUSIONIVCF implantation effectively prevents symptomatic and fatal PE of patients of lower limb and pelvic fractures with DVT in the peri-operative period.

Adult ; Aged ; Female ; Humans ; Leg Injuries ; complications ; Lower Extremity ; Male ; Middle Aged ; Pulmonary Embolism ; etiology ; prevention & control ; Retrospective Studies ; Vena Cava Filters ; Venous Thrombosis ; complications