Objective To analyze the death causes of the burn patients so as to explore the effective procedures which can raise burn management level. Methods One hundred patients died of burn injury during the past 20 years were enrolled in the study. The died patients were grouped as A (1984～1993) and B (1994～2003) groups, each group containing 50 cases. The mortality, burn area and depth, etiology, pre-hospital treatment, admission time, survival time, tracheostomy, the application of respirator and fibrobronchoscope, operation times, continuous renal replace treatment (CRRT), the incidence of inhalation injury and the pathogenesis of burn death were analyzed and compared between A and B groups. Results There were no differences in burn severity (area and degree), etiology and causes of burn death between the two groups. But the mortality in B group (1%) was evidently lower than that in A group (2%, P

Objective To investigate the cytotoxic effects and mechanism of PNP-CD chimeric gene vector originated from PNP/MeP-dR system on HCC cells. Methods The fusion suicide gene PNP-CD obtained by site directed mutagenesis technique was subcloned into pcDNA3.0 to construct a eukaryotic expression vector containing a chimeric gene, pcDNA3.0/ PNP-CD. After being identified by recombinant enzyme, PCR and subsequent sequencing, it was transfected into HepG2 cells by liposome-mediation method. The G418-resistant cellular clone with stable transfection of pcDNA3.0/PNP-CD, HepG2/PNP-CD was established by selection. The expression of PNP-CD gene was also verified by RT-PCR and Western blotting. The curve of cellular growth was assayed by Trypan blue exclusion. The cellular sensitivity of HepG2/PNP-CD to its specific prodrugs and its bystander effects were also assayed by MTT method. Results The chimeric gene, PNP-CD, was inserted into pcDNA3.0 correctly, and the stable expression of pcDNA3.0/PNP-CD in HepG2 cells was confirmed.This cellular clone was highly sensitive to its corresponding prodrugs. It was indicated that its bystander effects with the synergetic treatment of its specific prodrugs were substantially higher than those caused by the same vector with the administration of only a single prodrug, MeP-dR. Conclusion The bi-functional fusion suicide gene vector, pcDNA3.0/PNP-CD, yields powerful cytotoxic effects on HCC cells in the presence of the synergetic treatment of its specific prodrugs, which would be a high-performance therapeutic vector in gene therapy for liver cancer.

Objective To construct the eukaryotic expression vectors of fibroblast growth factor receptor 3(FGFR3) MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN, and to detect their expressions in human chronic myeloid leukemia(CML)K562 cell line.Methods The full-length FGFR3 (fgfr3-WT)and dominant negative FGFR3 (fgfr3-DN)were amplified by polymerase chain reaction (PCR). The two genes were respectively digested with EcoRⅠand BamHⅠ,and then ligated into MSCV/puro to construct the recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN which were tranduced into K562 cells by LipofectaminTM 2000 after PCR,double digestion and DNA sequencing.The expressions of FGFR3 protein in K562 cells were detected by Western blotting and flow cytometry. Results The recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN were amplified by PCR method, and the results showed fgfr3-WT of 2 400 bp and fgfr3-DN of 1 200 bp had been successfully cloned into MSCV-puro vector. The 2 400 bp fragment was oblained after double digestion of recombinant plasmid.The sequencing results showed that the size of fgfr3-WT was 2 400 bp which was the same as the sequence from GeneBank.Fgfr3-DN of 1 200 bp was also in conformity with the expected sequence.Compared with control (K562 MSCV)group,the expression level of FGFR3-WT in MSCV/puro-fgfr3-WT transfection (K562-WT)group was increased to above 10 times.There was high expression of FGFR3-DN in MSCV/puro-FGFR3-DN transfection (K562-DN)group,but there was no expressions in control(K562 MSCV)group and K562-WT group.The flow cytometry results showed that the high expressions of FGFR3-WT were in 57.5% cells in K562-DN group and the high expressions of FGFR3-DN were in 41.5% cells in K562-DN group. Conclusion The K562 cell lines highly expressing FGFR3-WT and FGFR3-DN are constructed successfully.

Objective To explore the curative effect and nursing of enhanced external counterpulsation (EECP) on patients with ischemic cerebral apoplexy. Methods In September 2013 to March 2013 in Shenzhen Futian District People′s Hospital neurology hospital, toally 286 patients with ischemic cerebral apoplexy were randomly divided into two groups, control group and treatment group with 143 cases in each group. The control group used conventional treatment and the treatment group used EECP. The score of clinical nerve function and defect improved Barthel index were compared. Result The score of clinical nerve function defect and improved Barthel index of the treatment group before the treatment were without difference (all P>0.05). The score of clinical nerve function defect and improved Barthel index of the treatment group were less than those of control group after the treatment (all P < 0.05). Conclusion Ischemic stroke patients with positive EECP can significantly increase clinical neural function and life ability , improve patient′s quality of life.

Objectives To examine the effects of enhanced external counterpulsation on arterial elasticity in stroke patients to provide clinical evidence for secondary prevention of patients with cerebral ischemic stroke. Methods Total 192 patients with ischemic stroke were enrolled and then divided into the EECP (n=107) and control (n=85) group. Auto-matic measurement synchronous atherosclerosis detector was use to measure brachial-ankle pulse wave velocity (BaP-WV) and cardio-ankle vascular index (CAVI). The difference of BaPWV and CAVI were evaluated before, at 36 hours and one month after EECP. Results The BaPWV and CAVI significantly decreased at 36 hours and 1 month after treat-ment in EECP groups compared to either pre-therapy or control groups (all P<0.05). Conclusions EECP can signifi-cantly reduce the BaPWV and CAVI and improve the arterial elasticity in patients with cerebral ischemic stroke. Thus, arterial elasticity may be an important index to evaluate the effects of EECP on cerebral ischemic stroke.

BACKGROUND:Pueraria is a member of isoflavone characterizing by decreasing the toxic effects of alcohol,relieving the effect of alcohol withdrawal,anti-oxidative effect,removing oxygen free radicals,and preventing cell injury induced by lipid peroxidation.OBJECTIVE:To investigate the inhibitory effect of puerarin on adipogenic differentiation of alcohol-induced human bone marrow mesenchymal stem cells (hBMSCs).DESIGN,TIME AND SETTING:An in vitro observation based on cytology was performed at Laboratory of Biochemistry and Molecular Biology,Basic Medical College of Zhengzhou University in October 2008.MATERIALS:The bone marrow was taken from the adult healthy volunteers from the Department of Orthopedics of the First Affiliated Hospital of Zhengzhou University.Puerarin standard was provided by the National Institute for the Control of Pharmaceutical and Biological Products.METHODS:The primary hBMSCs were purified from the adult marrow by gradient centrifugation.The second passaged cells were divided into 3 groups.Blank control group:Cells were cultured with DMEM culture media containing 10% fetal bovine serum;Model group:Cells were cultured with 0.09 mol/L alcohol;Experimental group:Cells were cultured with 0.09 mol/L alcohol and 10-6 mol/L puerarin;while,the culture liquid was changed every two or three days.Cells were cultured for 7 days in each group.MAIN OUTCOME MEASURES:PPARγ 2 gene expression with RT-PCR;amount of adipocyte with oil red "O" staining;alkaline phosphatase activity.RESULTS:PPAR-γ2 mRNA expression in the model group was significantly greater than experimental group and blank control group (P ＜0.05),while the expression in the experimental group was slightly greater than blank control group,but there was no significant difference (P ＞0.05).The number of adipocytes was the most in the model group,and the lipid droplet was large and abundant.The number of adipocytes in the experimental group was significantly less than model group (P ＜ 0.05),and there were few adipocytes in the blank control group.Compared with blank control group,alkaline phosphatase activity was decreased in the model group (P＜0.05);but the activity in the experimental group was increased compared to model group (P ＜ 0.05).CONCLUSION:Puerarin inhibits differentiation of hBMSCs into adipocytes induced by alcohol.

Objective To explore the therapeutic effect and prognosis of enhanced external counterpulsation (EECP)on acute cerebral ischemic stroke,to provide clinical evidence for the treatment of patients with acute cerebral ischemic stroke.Methods Total171 patients with acute cerebral ischemic stroke were enrolled and measured the NIHSS and mRS,before EECP,after36 hours EECP,and 3-month after attack.Then contrast the difference of these indicators.Result Compare with the control group,after EECP treatment and after 3-month attack,the scores of NIHSS were statistically significant,(after EECP:44.1％ vs 31.5％;after 3-month attack:55.6％ vs 40.5％),(P＜ 0.05).Compare with the control group,after 3-month attack,the score of mRS in EECP group was declined statistically significant,and the rate of favourable prognosis rise obviously (P＜0.05).Conclusion EECP can effectively improve neurological function and promote health and improve prognosis in the patients with acute cerebral ischemic stroke.

AIM: To investigate the protective effects and mechanisms of modified acidic fibroblast growth factor (maFGF) in anoxic reperfusion of rat hearts. METHODS: Using Langendorff apparatus, we established the model of anoxia/reperfusion of isolated hearts to compare the protective effects of maFGF and acidic fibroblast growth factor (aFGF). The changes of left ventricle development pressure (LVDP) and maximal rates of rise of ventricular pressure(dp/dt_~max ), maximal rates of decline of ventricular pressure (dp/dt_~min ) were determined, changes of LDH and MDA levels,SOD activity in efflux from coronary artery were also detected at different time point. RESULTS: Pretreatment with maFGF and aFGF produced a similar protective effect on myocardium during anoxia /reperfusion, including promoting obviously heart functional recovery after myocardial anoxia/reperfusion and reversing changes of LDH, MDA contents and SOD activity induced by anoxic/reperfusion. CONCLUSION: maFGF has a protective effect on anoxia/reperfusion heart, and the mechanism of this effect may be related to suppression of lipid peroxidation.

Objective:To analyze the risk factors of intracranial hemorrhage after implanting 125-iodine seeds for brain tumors.Methods:A total of 234 patients with intracranial tumors receiving treatment of 125-iodine seeds from March, 2013 to November, 2020 were retrospectively analyzed. Patients were divided into bleeding group and non-bleeding group according to whether postoperative intracranial hemorrhage was reported. Univariate and multivariate analysis was performed by logistic regression to determine the independent risk factors of intracranial hemorrhage.Result:A total of 22 cases (9.4%) reported postoperative intracranial hemorrhage in 234 patients treated with 125-iodine seeds. Univariate analysis showed that the type of tumor and the history of anti-angiogenic drug within one month were possible risk factors ( P<0.1). Multivariate logistic regression analysis showed that anti-angiogenic drug within one month was the independent risk factor for intracranial hemorrhage ( P<0.05). Conclusions:The application of anti-angiogenic drugs within one month is the independent risk factor of intracranial hemorrhage with 125-iodine seeds for the treatment of brain tumors.

BACKGROUND: Diabetic wounds are a major clinical challenge, because minor skin wounds can lead to chronic, unhealed ulcers and ultimately result in infection, gangrene, or even amputation. Studies on bone marrow derived mesenchymal stem cells (BMSCs) and a series of growth factors have revealed their many benefits for wound healing and regeneration. Platelet-rich plasma (PRP) may improve the environment for BMSC development and differentiation. However, whether combined use of BMSCs and PRP may be more effective for accelerating diabetic ulcer healing remains unclear. OBJECTIVE: We investigated the efficacy of BMSCs and PRP for the repair of refractory wound healing in a diabetic rat model. METHODS: Forty-eight rats with diabetes mellitus induced by streptozotocin were divided into four groups: treatment with BMSCs plus PRP, BMSCs alone, PRP alone, phosphate buffered saline. The rate of wound closure was quantified. A histopathological study was conducted regarding wound depth and the skin edge at 7, 14, and 28 days after surgery. RESULTS: Wound healing rates were significantly higher in the BMSC plus PRP group than in the other groups. The immunohistochemistry results showed that the expression of platelet/endothelial cell adhesion molecule 1, proliferating cell nuclear antigen, and transforming growth factor-beta1 increased significantly in the BMSC plus PRP group compared to the other treatment groups. On day 7, CD68 expression increased significantly in the wounds of the BMSC plus PRP group, but decreased markedly at day 14 compared to the controls. CONCLUSION: The combination of BMSCs and PRP aids diabetic wound repair and regeneration.
Amputation ; Animals ; Bone Marrow ; Cell Adhesion ; Diabetes Mellitus ; Gangrene ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; Mesenchymal Stromal Cells* ; Models, Animal ; Platelet-Rich Plasma* ; Proliferating Cell Nuclear Antigen ; Rats* ; Regeneration ; Skin ; Streptozocin ; Ulcer ; Wound Healing ; Wounds and Injuries