AIM: To investigate the effects of diazoxide, an ATP-sensitive K + channel opener on the ?-calpain activation, c-Fos and c- Jun expression in neonatal hypoxic-ischemic rat brain. METHODS:The animal model of hypoxic-ischemic brain injury (HIBI) was made in the 7-day -old SD rats. Diazoxide was injected into the left lateral ventricle prior or po st hypoxic-ischemia (HI) insults. Western blot was applied to detect the integra ted density (ID) of the nuclear c-Fos and c-Jun at 4h, and the cleavage of cytos olic ?-calpain at 24 h after HI insults. RESULTS: Low c-Fos and c-Jun expressions from cortical and hippocampal samples were observed in the tw o diazoxide groups, and significant differences in their expressions were found by comparison with the HI controls (P

Objective To study the neuroprotective effect of erythropoietin(EPO) on neonatal rats model with hypoxia-ischemia encephalopathy(HIE).Methods HIE was induced in rats on 7th day of postnatal age by ligation of right common carotid artery,followed by 2 h of hypoxia(80 mL/L O2).The subjects were divided into sham-operated group,control group and EPO group.EPO 4 000 U/(kg?day) was injected daily from day 2 pre-surgery for 9 to 16 days and PBS was injected in the control group.The neuroprotective effect of EPO on HIE model was detected by brain weight,the difference in weights between the ipsilateral(right) and contralateral(left) brain and the function test.In vitro study,the neural progenitor cell line C17.2 under gone apoptosis following an ischemia-like metabolic inhibition.The effect of EPO on the cell line ischemia modle 17.2 was evaluated by detecting Annexin V with flow cytometry.Results The signi-ficant and sustained brain injury in the hypoxia-ischemia and vehicle-treated group was observed and measured by reduction in relative weights of ipsilateral to contralateral and compromised sensorimotor functions in response to postural reflex test,compared with those of sham-operated animals(Pa

Objective To discuss the value and experience of the percutaneous vertebroplasty (PVP)in the treatment of vertebral body compression fracture(VCF)in aged osteoperosis.Methods PVP was performed in 44 cases with VCF including 28 with single vertebral compressed fracture,12 with double compressed fracture and four with triple compressed fracture,with 67 vertebrae,for clinical and radiologieal evaluation.Results The mean follow-up was 15 months(4-23 months).There could be seen immediate relief of pain in 40 cases,out-of-bed activities at operation day in 19 and out-of-bed activ- ities at second day after operation in 25.Postoperative X-ray showed uniformly distributed bone cement in the vertebral,without leakage.Conclusion PVP is a recommendable method for VCF,for it has ad- vantages of pain relief,vertebrae stabilization,minimal invasion and minor complications.

OBJECTIVE: To obtain the genetic polymorphism data of two STR loci D2S1399 and D5S2500 in Eastern Chinese Han population. METHODS: Blood samples or buccal swabs of unrelated Han individuals living in eastern China were analyzed using PCR-nature polyacrylamide gel electrophoresis-sliver staining method. RESULTS: 11 alleles of D2S1399 and 9 alleles of D5S2500 were observed in the samples respectively, the observed heterozygosity (Ho) values, the discrimination power (DP) values and the power of exclusion (PE) values of D2S1399 and D5S2500 is 0.745 and 0.807, 0.958 and 0.917, 0.554 and 0.643, respectively. CONCLUSION The result showed that D2S1399 and D5S2500 were highly informative loci and suitable for forensic application.
Alleles ; Asian People/genetics* ; Electrophoresis, Polyacrylamide Gel ; Forensic Medicine ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Silver Staining ; Tandem Repeat Sequences

OBJECTIVETo investigate the expression and clinical significance of EphA7 protein in primary hepatocellular carcinoma.

METHODSImmunohistochemistry and Western blot were used to detect the expression of EphA7 protein in 40 cases of primary hepatocellular carcinoma, their corresponding adjacent liver tissues and 10 cases of normal liver tissues. The relations with its clinical pathological parameters were analyzed too.

RESULTSExpression of EphA7 protein was mainly located in the cytoplasm and the blood vessels of the septa, which was found in hepatocellular carcinoma tissues, their corresponding adjacent liver tissues and normal liver tissues. Western blot analysis showed that the expression level of EphA7 protein in hepatocellular carcinoma (0.58 +/- 0.26) was greater than that in corresponding adjacent liver tissues (0.40 +/- 0.22, P < 0.05) and normal liver tissues (0.32 +/- 0.16, P < 0.05). But it had no significant difference between corresponding adjacent liver tissues and normal liver tissues (P > 0.05). EphA7 protein expression was correlated with histological differentiation, tumor thrombi in portal vein, lymph node metastasis and high AFP level (P < 0.05).

CONCLUSIONSEphA7 protein expression is significantly correlated with the biological behavior of primary hepatocellular carcinoma. The high expression of EphA7 protein may play an important role in the malignancy transformation, invasion progression and metastasis of primary hepatocellular carcinoma.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Receptor, EphA7 ; metabolism

OBJECTIVETo investigate the expression and clinical significance of Ephrin-A1 mRNA and protein expression level in hepatocellular carcinoma.

METHODSReverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry technique were used to detect the expression of the Ephrin-A1 mRNA and protein of 40 cases of hepatocellular carcinoma and their corresponding para-cancerous tissues and 10 cases of normal liver tissues. The relationships with its clinical pathology characters were analyzed.

RESULTSThe mRNA of Ephrin-A1 was expressed in all of the 40 cases of hepatocellular carcinoma and their corresponding para-cancerous tissues and 10 cases of normal liver tissues. Semiquantitative analysis showed that the mRNA expression level of Ephrin-A1 in hepatocellular carcinoma (0.5413 +/- 0.1527) was greater than that in corresponding para-cancerous tissues (0.3895 +/- 0.0549, P < 0.05) and normal liver tissues (0.3770 +/- 0.1055, P < 0.05); but between corresponding para-cancerous tissues (0.3895 +/- 0.0549) and normal liver tissues (0.3770 +/- 0.1055), the mRNA expression level had no significant difference (P > 0.05). The positive rates of Ephrin-A1 protein were 20% (2/10) in normal tissues, 35% (14/40) in para-cancerous tissues and 62% (25/40) in hepatocellular carcinoma tissues, respectively; the protein expression level of Ephrin-A1 was gradually rising (chi(2) = 14.762, P < 0.05). The overexpression of Ephrin-A1 protein was correlated with histological differentiation, tumor thrombi in portal vein and lymph node metastasis (P < 0.05).

CONCLUSIONSThe overexpression of Ephrin-A1 protein is correlated with histological differentiation, the lymph node metastasis and tumor thrombi in portal vein. It indicates that Ephrin-A1 may play an important role in the malignancy transformation, invasion progression and metastasis of hepatocellular carcinoma.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Differentiation ; Ephrin-A1 ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Metastasis ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To discuss a real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) wether if can be used to detect Brucella. Methods According to the BCSP31 gene sequences specific for Brucella, one pair of primers and one TaqMan probe were designed. A real-time PCR was developed with the BCSP31 fragments cloned into PMD18-T vector. The standard cure was established and the sensitivity, the species specificity and the stability of the assay were evaluated. The clinical blood specimens were detected by QT-PCR and compared with clinical diagnosis. Results The standard curve was established with the standard template and the relationship between the Ct and the DNA copy number was linear(r=0.999). The sensitivity of the real-time PCR was 5 copies/μl. The sensitivity of the common PCR was 5×102 copies/μl. The sensitivity was about 100 times higher than common PCR. Species specificity of this FQ-PCR assay evaluated using genomic DNA from 6 Bmcella strains and 5 non-Brucella strains and strong fluorescence was detected in all Brucella strains. The CV of intra-assay and inter-assay reproducibility were 0.71%,7.23%, reprectively. Twenty-four specimens from clinical brucellosis cases, 19 showed positive, the positive coincident rate was 79%(19/24). The negative results were obtained for all 31 negative control, and the negative coincident rate was 100%(31/31). Two were positive from all 30 specimens clinically suspected. Conclusions Highly specific, sensitive, repeatable and coincidental with clinic, this FQ-PCR is quite useful for rapid detection of tiny DNA of Brucella in various samples and laboratory diagnosis.

Objective To study the application value of dynamic monitoring of blood pressure in the prevention and treatment of elderly hypertension. Methods 519 hypertensive patients from December 2017 to December 2018 were monitored with ambulatory blood pressure monitoring, and were divided into the elderly group (≥60 years old, 264 cases) and the control group (<60 years old, 255 cases). The results of ambulatory blood pressure monitoring in two groups were analyzed, which inclued the circadian rhythm of blood pressure, blood pressure, pulse pressure, coefficient of variation of blood pressure, blood pressure load value, average heart rate and morning blood pressure surge. Results The incidence of abnormal circadian rhythm of ambulatory blood pressure in the elderly group was 76.5%. Compared with the control group, there were differences in the indexes of diastolic blood pressure (DBP), diastolic blood pressure load value (DBPLV), pulse pressure (PP), 24 h average heart rate (24 hAHR), systolic blood pressure coefficient of variation (SBPCV), 24 h diastolic blood pressure coefficient of variation (24 h DBPCV) and morning diastolic blood pressure surge (MDBPS) between the two groups(all P<0.05). There were differences in 24 h systolic blood pressure (24hSBP), night systolic blood pressure (nSBP), night diastolic blood pressure (nDBP), night pulse pressure (nPP), day systolic blood pressure load value (dSBPLV), ninght systolic blood pressure load value (nSBPLV), 24 h SBPCV, 24 h dDBPCV and other indicators among different blood pressure types in the elderly group (all P<0.05). Conclusion Ambulatory blood pressure monitoring indicators have important guiding value for the prevention and treatment of elderly hypertension.

BACKGROUNDLittle is known about neuronal death mechanisms following spinal cord ischemia. The present study aimed to investigate the protective effect of pentoxifylline (PTX) against spinal cord ischemia/reperfusion (I/R) injury.

METHODSRabbits sustained spinal cord ischemia following 45 minutes cross-clamping of the infrarenal aorta. Experimental groups were as follows: the first group of animals (sham, n = 8) underwent laparotomy alone and served as the sham group; the second group (I/R, n = 20) received carrier (3 ml saline solution) and served as the control group; the third group (PTX-A, n = 20) received PTX intravenously 10 minutes prior to ischemia; and the fourth group (PTX-B, n = 20) received PTX intravenously at the onset of reperfusion. Rabbits were evaluated for hind-limb motor function with the Tarlov scoring system at 48 hours. Serum was assayed with enzyme-linked immunosorbent assay for tumor necrosis factor alpha (TNF-alpha) and spinal cords were harvested for myeloperoxidase (MPO) activity, histopathological analysis, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling staining, platelet/endothelial cell adhesion molecule-1 (PECAM-1) and caspase-3 immunohistochemistry, and the number of necrotic and apoptotic neuron were counted and data analyzed at 12, 24, 48 and 72 hours of reperfusion. Spinal cords were studied by electron microscopy.

RESULTSImproved Tarlov scores were seen in PTX-treated rabbits as compared with ischemic control rabbits at 48 hours. A significant reduction was found in TNF-alpha in serum, activity of MPO and immunoreactivity of the PECAM-1 and caspase-3 in PTX-treated rabbits. There were fewer apoptotic neurons than necrotic neurons (P < 0.05). A significant decrease in both necrotic and apoptotic neurons was observed in the PTX-treated groups (PTX-A and PTX-B) compared with the I/R group (P < 0.05). Both necrotic and apoptotic neurons were found with the electron microscope.

CONCLUSIONSPTX may induce protection against ischemia injury in the spinal cord, thereby preventing both necrosis and apoptosis. A major mode of cell death in spinal cord ischemia/reperfusion injury is necrosis while apoptosis is not dominant.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Immunohistochemistry ; In Situ Nick-End Labeling ; Microscopy, Electron, Transmission ; Necrosis ; Pentoxifylline ; pharmacology ; therapeutic use ; Rabbits ; Reperfusion Injury ; prevention & control ; Spinal Cord ; blood supply ; pathology ; ultrastructure ; Spinal Cord Ischemia ; prevention & control ; Vasodilator Agents ; pharmacology ; therapeutic use

OBJECTIVEThe cascade of physiological events underlying hypoxic-ischemic brain damage (HIBD) remains to be fully established. The perinatal brain shows both an increased tolerance to hypoxic-ischemic (HI) injury and a faster and more complete recovery than the adult. It is, therefore, important to understand the sequence of events following hypoxia and ischemia in young animals. The present study aimed to clarify the time-course of the activation of the mu-calpain, and the expression of c-Fos, c-Jun, HSP70 and HSP27 proteins following severe HI (2 h hypoxia) and their relationship with each other.

METHODSA modified newborn rat model of HIBD that included a combination of hypoxia and ischemia as described by Rice was used. Forty-two postnatal 7-day-old Sprague-Dawley rats were randomly divided into seven groups (6 rats in each): 6 time-window groups and a normal control group. Samples were collected at 0, 1, 2, 4, 12 and 24 h after HI insults. The protein concentration was determined using a modified Bradford assay. mu-calpain activation, c-Fos, c-Jun, HSP70 and HSP27 expressions were observed respectively by Western blot from cortical and hippocampal samples.

RESULTSThe cleavage of cytosolic mu-calpain was observed from both cortical and hippocampal samples in neonatal rats after HI. The ratio 76:80 of mu-calpain was increased significantly post-HI and reached a maximum at 24 h in cortex and at 12 h in hippocampus after HI. The expressions of c-Fos and c-Jun from both cortical and hippocampal samples in neonatal rats were up-regulated and peaked at 2 or 4 h after HI, demonstrating significant differences at 1, 2, 4, and 12 h compared with that observed in the control (P < 0.05). When compared with that observed in cortex, the nuclear c-Fos expression from hippocampal samples was highly elevated at 2, 4 and 12 h but significantly decreased at 24 h after HI (P < 0.05), while the nuclear c-Jun expression from hippocampal samples was highly elevated at 0 and 1 h but significantly decreased at 4 and 24 h after HI (P < 0.05). Similarly, the expressions of HSP70 and HSP27 from both cortical and hippocampal samples were up-regulated and reached a maximum at 12 or 24 h after HI, demonstrating significant differences at 12 or 24 h both in cortex and hippocampus for HSP70, and at 24 h in cerebral cortex as well as at 12 and 24 h in hippocampus for HSP27 compared with the control (P < 0.05). Furthermore, in comparison with that observed in cortex, the HSP70 expression from hippocampal samples was highly elevated at 1 h, but significantly decreased at 4, 12 and 24 h after HI (P < 0.05), while the HSP27 expression was permanently elevated in hippocampus after HI.

CONCLUSIONThe neuronal injury induced by HI insults appears to involve many ongoing and simultaneous mechanisms. HI activates the calpains immediately, which may contribute to neuron apoptosis, and induces a significant brain neuroprotection, since there is an increased HSP70 expression and a relatively late remarkable HSP27 expression in hypoxic-ischemic neonatal rat brain. Nuclear c-Fos and c-Jun may participate in the pathogenesis of HIBD.

Animals ; Animals, Newborn ; Blotting, Western ; Brain ; metabolism ; pathology ; Calpain ; metabolism ; Enzyme Activation ; Female ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; metabolism ; Heat-Shock Proteins ; metabolism ; Hypoxia, Brain ; metabolism ; Male ; Neoplasm Proteins ; metabolism ; Proteins ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Proto-Oncogene Proteins c-jun ; metabolism ; Rats ; Rats, Sprague-Dawley ; Time Factors