Mechanism of Shikonin on spinal cord injury in rats based on TNFR/RIPK1 pathway / 中国骨伤

OBJECTIVE@#To explore the effect of shikonin on the recovery of nerve function after acute spinal cord injury(SCI) in rats.@*METHODS@#96 male Sprague-Dawley(SD)rats were divided into 4 groups randomly:sham operation group (Group A), sham operation+shikonin group (Group B), SCI+ DMSO(Group C), SCI+shikonin group (Group D).The acute SCI model of rats was made by clamp method in groups C and D . After subdural catheterization, no drug was given in group A. rats in groups B and D were injected with 100 mg·kg-1 of shikonin through catheter 30 min after modeling, and rats in group C were given with the same amount of DMSO, once a day until the time point of collection tissue. Basso-Beattie-Bresnahan(BBB) scores were performed on 8 rats in each group at 6, 12, and 3 d after moneling, and oblique plate tests were performed on 1, 3, 7 and 14 d after modeling, and then spinal cord tissues were collected. Eight rats were intraperitoneally injected with propidine iodide(PI) 1 h before sacrificed to detection PI positive cells at 24 h in each group. Eight rats were sacrificed in each group at 24 h after modeling, the spinal cord injury was observed by HE staining.The Nissl staining was used to observe survivor number of nerve cells. Western-blot technique was used to detect the expression levels of Bcl-2 protein and apoptosis related protein RIPK1.@*RESULTS@#After modeling, BBB scores were normal in group A and B, but in group C and D were significantly higher than those in group A and B. And the scores in group D were higher than those in group C in each time point (P<0.05). At 12 h after modeling, the PI red stained cells in group D were significantly reduced compared with that in group C, and the disintegration of neurons was alleviated(P<0.05). HE and Nissl staining showed nerve cells with normal morphology in group A and B at 24h after operation. The degree of SCI and the number of neuronal survival in group D were better than those in group C, the difference was statistically significant at 24h (P<0.05). The expression of Bcl-2 and RIPK1 proteins was very low in group A and B;The expression of RIPK1 was significantly increased in Group C and decreased in Group D, with a statistically significant difference (P<0.05);The expression of Bcl-2 protein in group D was significantly higher than that in group C (P<0.05).@*CONCLUSION@#Shikonin can alleviate the pathological changes after acute SCI in rats, improve the behavioral score, and promote the recovery of spinal nerve function. The specific mechanism may be related to the inhibition of TNFR/RIPK1 signaling pathway mediated necrotic apoptosis.

MiR-4484 regulates the expression of integrin α 6 in gastric cancer tissues and its significance / 中华肿瘤杂志

Objective: To investigate the relationship between the expression of integrin α 6 (ITGA6), miR-4484 and the pathologic stage of gastric cancer. Methods: Gastric cancer tissues and normal gastric mucosa tissues adjacent to cancer (>5 cm from tumor margin) of 30 patients with primary gastric cancer who underwent direct surgical resection without adjuvant therapy from June to September 2017 in West China Hospital of Sichuan University were selected. Real-time quantitative polymerase chain reaction (PCR) was used to detect the expression levels of miR-4484 and ITGA6, western blot was used to detect the expression level of ITGA6 protein, dual luciferase reporter gene was used to verify the relationship between ITGA6 and miR-4484. Spearman's correlation analysis was used to determine the relationship between miR-4484 and ITGA6 expression levels in gastric cancer tissues. Results: The expression level of ITGΑ6 in gastric cancer (32.30±13.47) was higher than that in matched normal gastric tissues (24.55±10.25, P=0.015), the area under the receiver operating characteristic (ROC) curve was 0.660 and the diagnostic sensitivity and specificity were 43.3% and 96.7%, respectively. The expression level of miR-4484 in gastric cancer (4.11±2.87) was lower than that of matched normal gastric tissues (5.75±2.80, P=0.029), the area under the ROC curve was 0.690 and the diagnostic sensitivity and specificity were 30.0% and 86.7%, respectively. The expression level of miR-4484 was negatively correlated with ITGA6 in gastric cancer tissues (r=-0.621, P<0.001). The expression level of ITGA6 protein in gastric cancer tissues (0.65±0.19) was higher than that in normal adjacent tissues (0.26±0.12, P<0.001). Compared with ITGA6 3'UTR wild-type+ miR-NC group, ITGA6 3'UTR wild-type+ miRNA mimics group had lower luciferase activity (50.69±5.10, 34.00±1.19, P<0.001), while the luciferase activity of ITGA6 3'UTR wild-type+ ASO miR-4484 group was higher than that of ITGA6 3'UTR wild-type+ miR-NC group (82.44±6.37, 50.69±5.10, P<0.001), indicated that ITGA6 was the direct target gene of miR-4484. The expression levels of miR-4484 in T1, T2, T3 and T4 (4a and 4b) gastric cancer tissues were 9.98±2.24, 5.28±2.03, 2.92±2.04 and 4.11±2.87, respectively, with statistical significance (P<0.001). The expression levels of ITGA6 in N0, N1, N2 and N3 gastric cancer tissues were 29.55±8.32, 21.71±3.75, 24.60±8.79 and 40.69±15.83, respectively, with statistical significance (P=0.022). The expression levels of miR-4484 in N0, N1, N2 and N3 gastric cancer tissues were 5.01±3.52, 5.48±2.76, 5.88±1.83 and 2.30±1.56, respectively, with statistical significance (P=0.032). The expression levels of ITGA6 in M0 and M1 gastric cancer tissues were 26.28±7.66 and 52.08±8.12, respectively, with statistical significance (P<0.001). The expression levels of miR-4484 in M0 and M1 gastric cancer tissues were 4.95±2.74 and 1.34±0.80, respectively, with statistical significance (P<0.001). Conclusions: ITGA6 is upregulated in gastric cancer tissues, while miR-4484 is downregulated in the gastric cancer group, and its expression level is related to the clinicopathological features of gastric cancer. ITGA6 is the direct target gene of miR-4484, implicates that miR-4484 may inhibit the invasion and metastasis of gastric cancer by regulating the expression of ITGA6. Both miR-4484 and ITGA6 may be the new prognostic markers and potential therapeutic targets of gastric cancer.

Observation on time-effect for the stimulating at Zusanli (ST 36) with minimally invasive embedding with PGLA in the healthy person / 中国针灸

OBJECTIVE@#To explore the objectivity and time-effect of stimulating effect at acupoint with PGLA in the healthy person, and to provide a basis for the rational interval of minimally invasive embedding of PGLA.@*METHODS@#Before embedding, 8 h, 3rd, 7th, 10th, 14th day after embedding, medical imaging magnetic resonance imaging (MRI) scanning technique was used to collect local T2WI pressure-lowering and T2-Mapping 8 echoes sequence image of left Zusanli (ST 36) in 8 cases of healthy person. The T2-Mapping 8 echoes sequence image was generated by the relevant software to the T2-Mapping image and the local T2 value was measured. The characteristics of local T2WI pressure-fat image signal intensity and the change of T2 value at left Zusanli (ST 36) with minimally invasive embedding with PGLA were observed and analyzed.@*RESULTS@#①There was no abnormal signal on the T2WI pressure-fat image on the left Zusanli (ST 36) point before the embedding. The high-signal was seen on the local T2WI pressure-fat image at each time point after embedding, there was no significant difference in local signal intensity between 8 h, 3rd and 7th day after embedding. The local signal intensity decreased on the 10th day after embedding, and the local signal intensity decreased significantly on the 14th day after embedding.②The T2 value at each time point after embedding increased significantly compared with that before embedding (all 0.05); there was no significant difference between the T2 value on the 7th and the 10th day after embedding (>0.05)，the T2 value on the 14th day after embedding was significantly lower than that on the 7th day after embedding (<0.01).@*CONCLUSION@#It has a stimulating effect on the local acupoints with minimally invasive embedding with PGLA in the healthy person, and the stimulating effect has certain time-effect. The effective stimulation time is about 2 weeks. The rational interval period for the minimally invasive embedding with the PGLA of the same specification type should be about 2 weeks.

The effect of comprehensive nursing intervention based on Comprehensive Geriatric Assessment on the life quality and pulmonary function in the rehabilitation of old patients with chronic obstructive pulmonary disease / 中国实用护理杂志

Objective To discuss the effect of comprehensive nursing intervention based on Comprehensive Geriatric Assessment (CGA) on the quality of life and pulmonary function in the rehabilitation of old patients with chronic obstructive pulmonary disease (COPD). Methods A total of 70 cases of COPD patients admitted from December 2013 to December 2016 in our hospital were randomly divided into the control group and CGA group, 35 cases in each group. The control group received routine nursing intervention, while the CGA group received comprehensive nursing intervention based on CGA assessment. The improvement of lung function and quality of life before and after nursing intervention was compared between the two groups. Results Before nursing intervention, the patients in CGA group and the control group showed no statistical significance in scores of each dimension of quality of life, such as daily life, social activities, anxiety, depression (P > 0.05). After nursing intervention, scores of each dimension of quality of life were significantly lower compared with before nursing intervention, and the quality of life score in CGA group were significantly higher than those of the control group: 1.81±0.21 vs.2.61±0.17, 1.51±0.29 vs. 1.97±0.34, 1.72±0.32 vs. 2.29±0.37, 1.91±0.23 vs. 2.86±0.48, each dimension score lower amplitude differences all had statistical significance(t=6.0898-17.5171, P<0.05). Nursing intervention before the CGA group of patients with the control group in patients with various lung function index [forced expiratory lung live(FVC), forced expiratory value 1 second(FEV1), peak expiratory flow and FEV1/ FVC] to compare differences had no statistical significance (P > 0.05). After nursing intervention, two groups of patients with the lung function of each index were significantly improved compared with before nursing intervention, the index of lung function and CGA group improved significantly better than control group:(2.80 ± 0.41)L vs.(2.24 ± 0.31)L,(1.98 ± 0.24)L vs.(1.50 ± 0.21)L, (70.31±10.21)％vs.(61.50±8.92)％,(4.49±0.28)％vs.(3.75±0.39)％, the differences were statistically significant(t=3.8444-9.1186, P<0.05). Conclusions Comprehensive nursing intervention based on CGA can effectively improve the pulmonary function of patients with COPD, improve the quality of life of patients, relieve anxiety and depression, and improve the effect of nursing intervention. It is worth popularizing in clinical practice.

Co-word clustering analysis for hot research areas of human caring based on PubMed / 中华护理杂志

Objective To identify hot research areas of nursing human caring in PubMed from 2012 to 2016, and to explore the present research status and development directions. Methods PubMed was searched using key words human caring. BICOMB 2.0 and SPSS 11.5 software were used to analyze high-frequency keywords and conduct co-word clustering analysis. Results We searched for 3088 related articles and extracted 42 high-frequency keywords (30.97%). Seven hot research areas were identified,including:human caring in nursing practice;nursing models,nursing theories;nursing education of human caring;hospice care;relationship between human caring and nurse-patient relation;human caring of cancer patients;family system,social support and human caring. Conclusion Analysis of research areas of nursing human caring in past 5 years is beneficial to understanding the present research status and development directions,and providing references for practice,research and education of human caring.

Association of nicotinamide adenine dinucleotide phosphate oxidase p22phox gene 549C>T polymorphism with coronary artery disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The p22phox is a critical component of the superoxide-generating vascular nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Several polymorphisms in p22phox gene are studied for their association with cardiovascular diseases. However, no publication is available to assess the relation of 549C > T polymorphism in p22phox gene to coronary artery disease (CAD) risk. This study was to investigate the effect of the p22phox gene 549C > T polymorphism on CAD risk.</p><p><b>METHODS</b>Hospital-based case-control study was conducted with 297 CAD patients and 343 healthy persons as the control group. Polymerase chain reaction and pyrosequencing using PSQ 96 MA Pyrosequencer (Biotage AB) were used to detect the polymorphisms. Multiple Logistic regression model was used to adjust the potential confounders and to estimate odds ratio (OR) with 95% confidence intervals (CIs).</p><p><b>RESULTS</b>The observed genotype frequencies of this polymorphism obeyed the Hardy-Weinberg equilibrium in both cases (P = 0.439) and controls (P = 0.668). The frequency of mutant genotypes (TT + CT) in cases (41.08%) was higher than that in controls (36.73%) with an OR = 1.20 (95%CI = 0.87-1.65). After the adjustment of the potential confounders, there was a significant association of the mutant genotypes with increased risk of CAD (OR = 1.57, 95%CI = 1.01-2.46, P = 0.047).</p><p><b>CONCLUSIONS</b>The mutant genotypes of the p22phox gene 549C > T polymorphism had a significant effect on the increased risk of CAD in this studied population.</p>

An improved liquid nitrogen freezing method for establishing canine models of femoral head necrosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish an canine model of femoral head osteonecrosis using an improved liquid nitrogen freezing method.</p><p><b>METHODS</b>Sixteen adult canines were divided into 4 groups at random and subjected to instant freezing of the unilateral femoral head with liquid nitrogen. The dogs were observed at 2, 4, 8, and 16 weeks after the operation for radiographic, gross and pathological changes of the femoral head.</p><p><b>RESULTS</b>Significant radiographic, gross and pathological changes occurred in the femoral head after the freezing.</p><p><b>CONCLUSION</b>Improved liquid nitrogen freezing of the femoral head provides a simple and convenient method for establishing animal models of femoral head osteonecrosis.</p>

Inhibition of growth and angiogenesis of U251 cell xenograft in vivo by short hairpin RNA targeting survivin gene / 中华外科杂志

<p><b>OBJECTIVE</b>To observe the impact of specific short hairpin RNA (shRNA) targeting survivin gene on tumorigenesis and angiogenesis of human brain glioblastoma U251 cells in vivo of nude mice.</p><p><b>METHODS</b>U251 cells, U251-SR cells transfected stably with shRNA eukaryotic expression vector pWH1-SR targeting survivin gene, and U251-P cells transfected stably with blank pWH1 vector, were inoculated respectively into subcutaneous tissue in flank of 15 nude mice (each group 5 mice), and the tumor growth status was observed and measured. Protein expressions of survivin, proliferating cell nuclear antigen (PCNA) and factor VIII related antigen (F VIII RAg) were investigated by immunohistochemistry SABC method, apoptotic cells were screened by TUNEL method, furthermore proliferative index (PI), apoptotic index (AI) and microvessel density (MVD) were measured respectively in each group of tumor specimens.</p><p><b>RESULTS</b>Comparing with those in U251 and U251-P groups, in U251-SR group, the tumorigenesis time delayed, tumor grew slowly, both tumor volume and tumor weight decreased significantly (P < 0.01 for both); Survivin protein expression was down-regulated markedly; PI and MVD decreased significantly, whereas AI increased remarkably (P < 0.01 for all).</p><p><b>CONCLUSIONS</b>The specific shRNA targeting survivin gene can inhibit significantly tumorigenesis and angiogenesis of U251 cells in vivo.</p>

Association between G894T mutation in endothelial nitric oxide synthase gene and premature coronary heart disease / 中华流行病学杂志

<p><b>OBJECTIVE</b>To assess the association between G894T (Glu298Asp) mutation in exon 7 of the endothelial nitric oxide synthase gene and premature coronary heart disease (P-CHD).</p><p><b>METHODS</b>Hospital-based case-control study was conducted. Newly-diagnosed CHD patients were recruited as study subjects. 132 CHD patients diagnosed at/before age 55 for males and 65 for females were assigned to P-CHD case group with other 172 CHD patients as the control group. Polymerase chain reaction with Ban II restriction enzyme digestion was performed to detect the G894T mutation.</p><p><b>RESULTS</b>G894T mutant genotypes in P-CHD group (TT, GT and GG frequencies were 6.06%, 20.45% and 73.48%, respectively) were significant higher than those in control group (TT, GT and GG frequencies were 1.74%, 11.63% and 86.63%, respectively) (P = 0.01). Mutant T allele frequency in P-CHD group was also significantly higher than that in control group (16.29% versus 7.56%, P = 0.001, OR = 2.38, 95% CI: 1.38 - 4.16). Stepwise multiple logistic regression analysis at 0.05 significant level with sex, smoking, alcohol drinking, and overweight covariates indicated that G894T mutation also having significant effect on P-CHD (P = 0.01, OR = 2.25, 95% CI: 1.19 - 4.26).</p><p><b>CONCLUSION</b>This study suggested that G894T mutation in endothelial nitric oxide synthase gene might serve as a major risk factor to the pathogenesis of P-CHD in this study population.</p>

Research on relationship of survivin gene expression with malignant proliferation and apoptosis of brain glioma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the expression level of inhibitor of apoptosis protein survivin gene in human brain glioma and its role in malignant proliferation and antiapoptosis of brain glioma.</p><p><b>METHODS</b>Eighty-three cases of brain glioma specimen was chosen, protein expression of survivin and proliferating cell nuclear antigen (PCNA) was investigated by immunohistochemistry streptavidin-biotin complex (SABC) method, the immunoreactivity score (IRS) of survivin and the proliferative index (PI) were counted. Apoptotic cells were screened by TdT-mediated dUTP-biotin nick-end labeling (TUNEL) method, and the apoptotic index (AI) of brain glioma was calculated.</p><p><b>RESULTS</b>The survivin IRS, PI and AI of brain glioma were 3.8 +/- 3.9, (28.4 +/- 19.5)% and (1.0 +/- 0.8)% respectively, and all of them were elevated with the increase of pathological grade of brain glioma (P < 0.01 for all). PI in survivin positive group was significantly higher than that in survivin negative group (P < 0.01), and PI was positively correlated with survivin IRS (r = 0.740, P < 0.01). There was no significant difference between AI in survivin positive group and that in survivin negative group (P > 0.05), however, AI was negatively correlated with survivin IRS (r = -0.307, P < 0.01).</p><p><b>CONCLUSIONS</b>Survivin is overexpressed in brain glioma, and which may play important roles in malignant proliferation and antiapoptosis of brain glioma.</p>

Construction and biological activity study of human osteoprotegerin expressing adenoviral system / 生物工程学报

Using the isolated total RNA from osteosacoma cell line MG63, the cDNA encoding human OPG was amplified by RT-PCR. A recombinant adenoviral vector carrying cDNA of OPG was constructed and OPG expression in mouse myoblast C2C12 cells was confirmed by Western blot and ELISA. The secreted expression of OPG protein persisted more than 6 weeks in vitro, and the growth of C2C12 cells infected by recombinant adenoviral were in good state. Osteoclasts derived from mouse bone marrow cells infected with recombinant adenoviral made less number of TRAP positive cells and resorption pits formed on dentine slices.