Objective To evaluate the transfection efficiency and expression level of hepatocyte growth factor (HGF) by transfecting a recombinant adenovirus carrying HGF gene (Ad-HGF) into bone marrow mesenchymal stem cells (BMSCs) and to explore the effect of the expression supernatant on BMSCs in vitro so as to lay a foundation for the manufacture of gene medicine which expresses efficient cell factors. Methods Rat BMSCs were isolated using Percoll density gradient method and cultured according to the adherent property of BMSCs. The expression of c-Met was detected by immunohistochemical examination. BMSCs were infected with a recombinant adenovirus carrying green fluorescent protein gene (Ad-GFP) at multipl icity of infection (MOI, 0, 25, 50, 100, and 200 pfu/cell). To select an optimal MOI, the transfection efficiency and the degree of cell damage were assayed by flow cytometry and MTT, respectively, at 48 hours after transfecting. The expression of HGF in BMSCs transfected with optimal MOI Ad-HGF was measured with ELISA assay. MTT method was used to evaluate the prol iferation effect of HGF expression supernatant on BMSCs. Results Immunohistochemical staining showed that BMSCs expressed c-Met receptor for HGF. At 48 hours after transfecting with different MOI Ad-GFP (0, 25, 50, 100, and 200 pfu/cell), the transfection efficiencies were 0.34% ± 0.04%, 40.72% ± 0.81%, 61.72% ± 1.04%, 85.33% ± 0.83%, and 17.91% ± 0.63%, respectively; and the highest transfection efficiency was observed at 100 pfu/cell MOI. The cell damage was obviously observed when MOI was 200 pfu/cell. The expression of HGF in BMSCs reached the highest level after being transfected with 100 pfu/cell MOI Ad-HGF for 48 hours. The expression product could stimulate the prol iferation of BMSCs. The prol iferation of BMSCs gradually rose with the increase of HGF protein, and reached the highest level at 10% (320 pg). Conclusion BMSCs can be transfected efficiently with Ad-HGF and express HGF protein, which stimulates the prol iferation of BMSCs. It suggests that BMSCs is an ideal repair cells with gene vector.
Objective To explore the clinical application of tissue flap in repairing the residual cavity formed by breast conserving surgery. Methods Retrospective analysis for clinical data of 120 early breast cancer patients who received breast conserving surgery in our hospital from January 2013 to December 2016 was performed. Among them, 60 cases (observation group) were implemented tissue flap repairing operation while the other 60 cases (control group) were operated by using the traditional surgery method. The clinical data of the two groups were compared analytically in four aspects: postoperative complications, postoperative breast beauty, subjective satisfaction of patients, and postoperative recurrence and metastasis. Results ① Postoperative complications: 3 cases of complications occurred in the control groupand 2 cases in the observation group, and there was no significant difference in the incidence of complications between the 2 groups (χ2=0.209, P=0.648). ② Postoperative breast beauty: in the control group, there were 23 excellent cases, 16 good cases, 12 common cases, and 9 poor cases; in the observation group, there were 51 excellent cases, 5 good cases, 3 common cases, and 1 poor cases. The difference of postoperative breast beauty between the 2 groups was statistically significant (Z=–5.234, P<0.001). ③ Subjective satisfaction of patients: in the control group, 28 cases very satisfied, 18 cases satisfied, 12 cases generally satisfied, and 2 cases dissatisfied; in the observation group, 40 cases satisfied, 18 cases very satisfied, and 2 cases generally satisfied. The subjective satisfaction of the patients in the observation group was better than that of the control group (Z=–4.381, P<0.001). ④ Postoperative follow-up: no recurrence and death occurred in both of 2 groups, but 2 cases occurred metastasis in the control group and 1 case in the observation group. The contrast difference in the incidence of metastasis was not statistically significant (χ2=0.342, P=0.559). Conclusion During breast conserving surgery, the use of surrounding tissue flap to repair residual cavity, compared with traditional breast conserving surgery without tissue flap repairment, does not increase postoperative complications, recurrence, and metastasis, and it has good cosmetic effect after breast surgery, as well as the patients’ subjective satisfaction is good.
Objective To explore the effects of bone marrow mesenchymal stem cells (BMSCs) transfected with adenovirus hepatocyte growth factor (Ad-HGF) on wound repair in diabetic rats. Methods BMSCs from male Wistar rats were isolated by density gradient centrifugation, cultured, and transfected with Ad-HGF. The multi pl icity of infection was 100. Diabetic models were establ ished in 20 female Wistar rats by diets in high fat and sugar plus intraperitoneal injection ofstreptozotocin (30 mg/kg). Then 2 full-thickness skin wounds (approximately 1.5 cm in diameter) were made on the dorsum. The rats were randomly divided into 4 groups (n=5 rats). After wounding, the 0.3 mL suspensions of BMSCs (group A), Ad- HGF (group B), BMSCs transfected with Ad-HGF (group C), and PBS (group D) were injected directly into the derma of wounds. The transverse diameter and longitudinal diameter of wound were measured at 21 days after treatment. At 7 days and 28 days after treatment, HE staining was performed to evaluate wound heal ing. The contents of hydroxyprol ine and advanced glycosylation end products (AGEs) in the wounds were measured by enzyme l inked immunosorbent assay and fluorospectrophotometer, respectively, at 3, 7, 14, and 28 days after treatment. Results At 21 days after treatment, the wounds almost healed in group C, and the transverse diameter and longitudinal diameter were 0 and (0.110 ± 0.024) cm, respectively. But the wounds healed partially in groups A, B, and D, and the transverse diameter and longitudinal diameter were (0.470 ± 0.051) cm and (0.590 ± 0.041) cm, (0.390 ± 0.042) cm and (0.480 ± 0.032) cm, and (0.700 ± 0.068) cm and (0.820 ± 0.068) cm, respectively. There were significant differences in wound heal ing between group C and groups A, B, and D (P lt; 0.05). The wound heal ing time of group C [(20.5 ± 1.9) days] was significantly shorter (P lt; 0.05) than those of groups A, B, and D [(28.3 ± 1.9), (25.9 ± 2.3), and (36.6 ± 5.1) days]. At 7 days, the HE staining showed that evident epidermis transportation, collagen formation, and leukocytes infiltration were observed in group C. At 28 days, the HE staining showed that the epidermis in group C was significantly thinner and more regular than those in other groups, and the decreased collagen and many small vessels were observed in group C. The content of hydroxyprol ine in group C was higher than those in groups A, B, and D at 7 days and 14 days (P lt; 0.05). The contents of AGEs in group C was lower than those in groups A, B, and D at 14 days and 28 days (P lt; 0.05). Conclusion Transplantation of BMSCs transfected with Ad-HGF can accelerate the wounds repair in diabetic rats.
ObjectiveTo study the protective effect and mechanism of ophiopogonin D (OP-D) on lipopolysaccharide induced acute lung injury (ALI) in mice.MethodsFifty SPF C57BL/6 mice were randomly divided into five groups, ie. a control group, a sham operation group, a model group, an OP-D group (10 mg·kg–1·d–1), and a dexamethasone group (2 mg·kg–1·d–1), with 10 mice in each group. One day before the establishment of the model, the OP-D group and the dexamethasone group received the corresponding drugs by gavage. The model group, the OP-D group and the dexamethasone group received lipopolysaccharide (2 mg/kg, 30 μL) through the trachea to establish the ALI model. The sham operation group received the same volume of normal saline. The blank control group was not treated. Six hours after the operation, the mice were weighed and then killed for peripheral blood and lung tissue. The weight of lung tissue was measured to evaluate the degree of pulmonary edema; the pathological changes of lung tissue were observed by hematoxylin-eosin staining; the mRNA expressions of interleukin (IL)-6, IL-10, and IL-17 in lung tissue were detected by qPCR; the percentage of Th17 and Treg cells in peripheral blood was detected by flow cytometry.ResultsCompared with the model group, the degree of pulmonary edema in the OP-D group decreased significantly (P<0.05), the lung tissue injury decreased, the mRNA expressions of IL-6 and IL-17 in the lung tissue and the proportion of Th17 cells in the peripheral blood decreased significantly (P<0.05), the proportion of Treg cells in the peripheral blood and the mRNA expression of IL-10 in the lung tissue increased significantly (P<0.05).ConclusionOP-D may have therapeutic effect on LPS induced ALI in mice by regulating the balance of Th17/Treg cells.
ObjectiveThis study was aimed to evaluate the clinical efficacy of mechanical thrombectomy using the AngioJet System for the treatment of lower extremity acute arterial embolism and thrombosis.MethodsThe clinical data of 20 patients with acute lower extremity arterial embolism and thrombosis admitted to the Department of Vascular Surgery in the People’s Hospital in Gansu Province where the author worked from September 2016 to March 2017, were retrospectively analyzed. All patients were treated with the AngioJet mechanical thrombectomy system. Clinical data of the patients were retrospectively collected. The clinical efficacy of AngioJet mechanical thrombectomy wasanalyzed.ResultsEighteen (90.0%) of the 20 patients successfully completed the mechanical thrombectomy by using the AngioJet System. The mean time for hospital stay and operation was (4.2±1.4) d and (1.3±0.4) h, respectively. The average doses of urokinase and heparin during operation were (35.80±12.30) ×104 U and (45.10±8.30) mg, respectively. Two patients received a complementary treatment of incision for removing the thrombus. Two patients received catheter-directed thrombolysis after the mechanical thrombectomy, 5 patients received bare-metal stent implantation after balloon expansion. Clinical success was in 16 cases. According to the Cooley standard, 10 patients were in excellent condition,6 in good condition, 2 in fair condition, and 2 in poor condition. There were 2 cases of distal arterial embolization,2 cases of antecardial discomfort of bradycardia, and 4 cases of bleeding at the puncture point, but no serious bleeding complications such as gastrointestinal and intracranial hemorrhage occurred. A total of 16 patients presented myoglobinuria during and after operation. All patients were followed up for 6–12 months. The results of ultrasound examination showed that the artery was patency in 15 cases. One patient died of myocardial infarction in 9 months after surgery,2 patients developed lower extremity ischemia symptoms again after surgery, and 2 patients had lower extremity ulcer caused by lower extremity ischemia symptoms. During the follow-up period, no lower limb necrosis, amputation, and death occurred in the remaining patients.ConclusionsThe AngioJet mechanical thrombectomy system is safe and effective. Combined with the use of catheter-directed thrombolysis and stent implantation, the AngioJet mechanical thrombectomy could lead to quick recovery of the perfusion of the lower extremity and improve the limb salvage rates, exhibiting excellent clinical value.
ObjectiveTo evaluate the efficacy and safety of intracavitary treatment for iliac vein compression syndrome（IVCS）with acute lower extremity deep venous thrombosis (DVT).MethodsThe clinical data of 57 patients with IVCS and lower extremity DVT, who undergoing with stent implantation, balloon expansion and Angiojet rheolytic thrombectomy from June 2015 to June 2018, were retrospectively analyzed. The effect of treatment was evaluated by the changes of thigh circumference difference between the affected side and the healthy side, and the thrombosis clearance rate in the operating. In addition, the incidence of post-thrombotic syndrome (PTS) and stent patency rate were analyzed after long-term follow-up based on the change of Villaita scale score and ultrasound examination of lower extremity veins.ResultsThe success rate of surgical technique was 100%, and there was no pulmonary embolism during operating and postoperative. Lower extremity deep vein thrombosis clearance levels Ⅲ 48 cases (84.2%), Ⅱ 9 cases (15.8%), the changes of thigh circumference difference between the affected side and the healthy side from preoperative (5.8±1.7) cm to (3.7±1.0) cm. One year follow-up after operation, the primary patency rate of stent was 86.0% and PTS occurred in 8 patients (14.0%).ConclusionStent implantation, balloon expansion and Angiojet rheolytic thrombectomy for IVCS with acute lower extremity DVT is a safe, effective with low incidence of complications and efficient thrombus clearance.