Protective effect of thodioloside and bone marrow mesenchymal stem cells infected with HIF-1-expressing adenovirus on acute spinal cord injury

Rhodioloside has been shown to protect cells from hypoxia injury,and bone marrow mesenchymal stem cells have a good effect on tissue repair.To study the effects of rhodioloside and bone marrow mesenchymal stem cells on spinal cord injury,a rat model of spinal cord injury was established using the Infinite Horizons method.After establishing the model,the rats were randomly divided into five groups.Rats in the control group were intragastrically injected with phosphate buffered saline(PBS)(5μL).PBS was injected at 6 equidistant points around 5 mm from the injury site and at a depth of 5 mm.Rats in the rhodioloside group were intragastrically injected with rhodioloside(5 g/kg)and intramuscularly injected with PBS.Rats in the mesenchymal stem cell(MSC)group were intramuscularly injected with PBS and intramuscularly with MSCs(8×10^6/mL in a 50-μL cell suspension).Rats in the Ad-HIF-MSC group were intragastrically injected with PBS and intramuscularly injected with HIF-1 adenovirus-infected MSCs.Rats in the rhodioloside+Ad-HIF-MSC group were intramuscularly injected with MSCs infected with the HIF-1 adenovirus and intragastrically injected with rhodioloside.One week after treatment,exercise recovery was evaluated with a modified combined behavioral score scale.Hematoxylin-eosin staining and Pischingert’s methylene blue staining were used to detect any histological or pathological changes in spinal cord tissue.Levels of adenovirus IX and Sry mRNA were detected by real-time quantitative polymerase chain reaction and used to determine the number of adenovirus and mesenchymal stem cells that were transfected into the spinal cord.Immunohistochemical staining was applied to detect HIF-1 protein levels in the spinal cord.The results showed that:(1)compared with the other groups,the rhodioloside+Ad-HIF-MSC group exhibited the highest combined behavioral score(P<0.05),the most recovered tissue,and the greatest number of neurons,as indicated by Pischingert’s methylene blue staining.(2)Compared with the PBS group,HIF-1 protein expression was greater in the rhodioloside group(P<0.05).(3)Compared with the Ad-HIF-MSC group,Sry mRNA levels were higher in the rhodioloside+Ad-HIF-MSC group(P<0.05).These results confirm that rhodioloside combined with bone marrow mesenchymal stem cells can promote the recovery of spinal cord injury and activate the HIF-1 pathway to promote the survival of bone marrow mesenchymal stem cells and repair damaged neurons within spinal cord tissue.This experiment was approved by the Animal Ethics Committee of Gansu University of Traditional Chinese Medicine,China(approval No.2015KYLL029)in June 2015.

Keratinocyte growth factor gene therapy ameliorates ulcerative colitis in rats

AIM:To investigate the effect of keratinocyte growth factor(KGF) gene therapy in acetic acid-induced ulcerative colitis in rat model.METHODS:The colitis of Sprague-Dawley rats was induced by intrarectal infusion of 1 mL 5%(v/v) acetic acid.Twenty-four hours after exposed to acetic acid,rats were divided into three experimental groups:control group,attenuated Salmonella typhimurium Ty21a strain(SP) group and SP strain carrying human KGF gene(SPK) group,and they were separately administered orally with 10% NaHCO3,SP or SPK.Animals were sacrificed and colonic tissues were harvested respectively on day 3,5,7 and 10 after administration.Weights of rats,colonic weight/length ratio and stool score were evaluated.Histological changes of colonic tissues were examined by hematoxylin and eosin(HE) staining method.The expression of KGF,KGF receptor(KGFR) and TNF-α were measured either by enzyme-linked immunosorbent assay or Western blotting.Immunohistochemistry was used to detect the cellular localization of KGFR and Ki67.In addition,superoxide dismutase(SOD) activity and malondialdehyde(MDA) contents in the homogenate were measured.RESULTS:Body weight and colonic weight/length ratio were declined in SPK group compared with SP and control groups(body weight:272.78 ± 17.92 g vs 243.72 ± 14.02 g and 240.68 ± 12.63 g,P < 0.01;colonic weight/length ratio:115.76 ± 7.47 vs 150.32 ± 5.99 and 153.67 ± 5.50 mg/cm,P < 0.01).Moreover,pathological changes of damaged colon were improved in SPK group as well.After administration of SPK strain,KGF expression increased markedly from the 3rd d,and remained at a high level till the 10th d.Furthermore,KGFR expression and Ki67 expression elevated,whereas TNF-α expression was inhibited in SPK group.In the group administered with SPK,SOD activity increased significantly(d 5:26.18 ± 5.84 vs 18.12 ± 3.30 and 18.79 ± 4.74 U/mg,P < 0.01;d 7:35.48 ± 3.35 vs 22.57 ± 3.44 and 21.69 ± 3.94 U/mg,P < 0.01;d 10:46.10 ± 6.23 vs 25.35 ± 4.76 and 27.82 ± 6.42 U/mg,P < 0.01) and MDA contents decreased accordingly(d 7:7.40 ± 0.88 vs 9.81 ± 1.21 and 10.45 ± 1.40 nmol/mg,P < 0.01;d 10:4.36 ± 0.62 vs 8.41 ± 0.92 and 8.71 ± 1.27 nmol/mg,P < 0.01),compared with SP and control groups.CONCLUSION:KGF gene therapy mediated by attenuated Salmonella ameliorates ulcerative colitis induced by acetic acids,and it may be a safe and effective treatment for ulcerative colitis.

Endothelial progenitor cells in peripheral blood may serve as a biological marker to predict severe acute pancreatitis

AIM To investigate the significance of endothelial progenitor cells(EPCs) in predicting severe acute pancreatitis(SAP).METHODS We recruited 71 patients with acute pancreatitis(AP) and excluded 11 of them; finally, cases of mild acute pancreatitis(MAP)(n = 30) and SAP(n = 30), and healthy volunteers(n = 20) were internalized to investigate levels of EPCs, C-reactive protein(CRP), tumor necrosis factor-alpha(TNF-α), fibrinogen(FIB)and white blood cells(WBC) in peripheral blood. RESULTS The levels of TNF-α, WBC, FIB and CRP were higher both in SAP and MAP cases than in healthy volunteers(P < 0.05, all). Interestingly, the level of EPCs was higher in SAP than MAP(1.63% ± 1.47% vs 6.61% ± 4.28%, P < 0.01), but there was no significant difference between the MAP cases and healthy volunteers(1.63% ± 1.47% vs 0.55% ± 0.54%, P > 0.05). Receiver operating characteristics curve(ROC) showed that EPCs, TNF-α, CRP and FIB were significantly associated with SAP, especially EPCs and CRP were optimal predictive markers of SAP. When the cut-off point for EPCs and CRP were 2.26% and 5.94 mg/d L, the sensitivities were 90.0% and 73.3%, and the specificities were 83.3% and 96.7%. Although, CRP had the highest specificity, and EPCs had the highest sensitivity and highest area under the curve value(0.93). CONCLUSION Data suggest that EPCs may be a new biological marker in predicting SAP.

Human Papillomavirus 16E6 Oncogene Mutation in Cervical Cancer

Objective： Cervical cancer （CC） is the second most common type of cancer in women worldwide, after breast cancer. High-risk human papillomaviruses （HR-HPVs） are considered to be the major causes of cervical cancer. HPV16 is the most common type of HR-HPVs and HPV16 E6 gene is one of the major oncogenes. Specific mutations are considered as dangerous factors causing CC. This study was designed to find mutations of HPV16 E6 and the relationship between the mutations and the happening of CC. Methods： The tissue DNA was extracted from 15 biopsies of CC. Part of HPV16 E6 gene （nucleotide 201-523） was amplified by polymerase chain reaction （PCR） from the CC tissue DNA. The PCR fragments were sequenced and analyzed. Results： The result of PCR showed that the positive rate of HPV16 E6 was 93.33% （14/13）. After sequencing ana analyzing, in the 13 out of 14 PCR fragments, 4 maintained prototype （30.77%）, 8 had a same 350G mutation （61.54%）, and 1 had a 249G mutation （7.69%）. Conclusion： This study suggest that there is a high infection rate of HPV in cervical cancer and most of the HPV16 E6 gene has mutations. Those mutations may have an association with the development of cervical cancer.

Potential of Sestrin2 in the prevention and treatment of chronic diseases

The prevalence and lethality of chronic non-communicable diseases are constantly rising,becoming a global public health problem.The occurrence and development of chronic diseases are closely related to the generation of excessive free radicals and active oxygen in the body,and anti-oxidation will become an effective treatment.Sestrin2 protein,as a new stress protein found in mammals in recent years,has unique advantages in antioxidants,and is expected to become an effective biomarker and therapeutic target for chronic diseases.The following is a review of the regulatory role and mechanism of sestrin2 in chronic diseases in order to provide a reference for the research of other scholars.

Research status of association between serum minerals and complications of type 2 diabetes

As a chronic metabolic disease,type 2 diabetes mellitus is prone to various microvascular and macrovascular complications.As a micronutrient,minerals have a small amount of body demand,but they are essential to the body's metabolism and play an important role in the occurrence of diabetes complication.This review summarizes the relationship between serum calcium,magnesium,iron,copper,and zinc and the complications of type 2 diabetes mellitus.It provides references for related scholars,and also provide favorable basis for early diagnosis,nutrition management with prevention and treatment of complications of type 2 diabetes.