AIMTo study the function of c-Jun N-terminal kinase 3 (JNK3) in the process of ischemic/reperfused heart injury and the mechanism underlying the protective action of magnesium lithospermate B (MTB), a bioactive compound isolated from Danshen.

METHODSBy in situ hybridization, JNK3 mRNA was detected in the ventricular preparations of the Langendorff ischemic/reperfused rat heart. The inhibitory effect of MTB on the expression of JNK3 mRNA was also investigated.

RESULTSThe purple and blue hybridization signals were located in the cytoplasm of the cardiomyocytes, which were weaker in the non-perfused hearts and stronger in the hearts encountered 30 min of ischemia and 30 min of reperfusion. Image analysis showed that the expression of JNK3 mRNA in the cardiomyocytes increased after 30 min of ischemia and 30 min of reperfusion, which showed significant difference compared with that in the cardiomyocytes of the non-perfused heart and the control heart (P < 0.05). Treatment with of 0.1, 1 and 10 mumol.L-1 MTB abolished the elevation of JNK3 mRNA expression in the ischemic/reperfused heart (P < 0.05).

CONCLUSIONJNK3 may be another component in the signal transduction pathway of ischemia/reperfusion induced cardiomyocyte apoptosis. MTB may protect the heart from ischemia/reperfusion injury by reducing apoptosis through inhibition of the JNK3 activity.

Animals ; Cardiotonic Agents ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; Male ; Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; genetics ; metabolism ; Myocardial Ischemia ; complications ; Myocardial Reperfusion Injury ; enzymology ; etiology ; pathology ; Myocytes, Cardiac ; drug effects ; enzymology ; RNA, Messenger ; biosynthesis ; drug effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the association of blood lipids with the development, clinical stage, allergic condition, and pulmonary function of asthma.

METHODSA total of 56 children with asthma who attended the hospital between October 2016 and March 2017 were enrolled as the asthma group, and 46 children who underwent physical examination as the healthy control group. According to the clinical manifestations, the children with asthma were divided into acute exacerbation group (n=24) and chronic persistent group (n=32). According to the results of skin prick test (SPT) and serum IgE measurement, the children with asthma were divided into non-allergic asthma group (n=16) and allergic asthma group (n=38). Fasting blood lipid levels were measured in both asthma and control groups. Pulmonary function tests were performed for asthmatic children.

RESULTSThere were no significant differences in blood lipid levels between the asthma and control groups (P>0.05). The acute exacerbation group had significantly lower serum levels of high-density lipoprotein (HDL) and total cholesterol compared with the control group and the chronic persistent group (P<0.05). The allergic asthma group had a significantly lower serum HDL level than the non-allergic asthma group (P<0.05). In asthmatic children aged 6-13 years, the ratios of the measured values to the predicted values for forced vital capacity, peak expiratory flow, and maximal expiratory flow at 50% of vital capacity had a linear regression relationship with HDL and were positively correlated with HDL (P<0.05). Forced expiratory volume in one second and maximal mid-expiratory flow had a linear regression relationship with both HDL and LDL and were positively correlated with them (P<0.05).

CONCLUSIONSBlood lipids are associated with the clinical stage, allergic condition, and lung function of childhood asthma. This indicates that blood lipids may be involved in several aspects of the pathogenesis of childhood asthma.

Adolescent ; Asthma ; blood ; physiopathology ; Child ; Child, Preschool ; Female ; Forced Expiratory Volume ; Humans ; Lipids ; blood ; Lung ; physiopathology ; Male ; Vital Capacity

OBJECTIVETo evaluate the clinical effects of transpedicular eggshell technique in treating thoracolumbar deformity.

METHODSFrom December 2008 to December 2011,36 patients with thoracolumbar deformity were treated with transpedicular eggshell technique. There were 20 males and 16 females with an average age of 45 years old (ranged from 20 to 58). Among them, 5 cases were congenital hemivertebrae deformity, 12 cases were secondary to tuberculotic deformity, 14 cases were post-traumatic deformity with pain, 5 cases were ankylosing spondylitis. Low back pain, living ability, scoliotic Cobb angle were analyzed according to VAS scoring, Oswestry Disability Index (ODI), radiological examination.

RESULTSAverage operative time was 245 min and average bleeding was 1 900 ml in 36 patients. All patients were followed up more than 1 year and obtained bone fusion at 1 year after operation. Preoperative,postoperative at 1 week and 1 year, VAS scoring was 7.2 +/- 1.4, 2.5 +/- 1.0, 1.8 +/- 0.5, respectively; ODI was (72.50 +/- 10.80)%, (42.50 +/- 11.10)%, (22.50 +/- 7.90)%, respectively; kyphosis Cobb angle was (76.31 +/- 2.52) degrees, (23.66 +/- 1.16) degrees, (23.67 +/- 1.16) degrees, respectively; lumbar scoliosis Cobb angle was (71.86 +/- 4.02) degrees, (30.81 +/- 2.33) degrees, (30.82 +/- 2.32) degrees, respectively. Postoperative at 1 week and 1 year,above data had obviously improved than that of preoperative (P < 0.05); and there was no significant difference in Cobb angle between postoperative at 1 week and postoperative at 1 year (P > 0.05).

CONCLUSIONTreatment of thoracolumbar deformity with transpedicular eggshell technique could obtain effective correcting and clinical results.

Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Orthopedic Procedures ; methods ; Thoracic Vertebrae ; abnormalities ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo evaluate the utility of the cross-species screening strategy for investigating key molecule(s) involved in onset and progression of hepatocellular carcinoma (HCC).

METHODSHCC-related molecule data from our previous studies and in the literature were collected to establish a cross-species dataset. Tissue samples of HCC, non-HCC surrounding liver (para-HCC), and normal liver that were collected from humans, tree shrews and rats. The genes reported to have the most differential expression in HCC were verified by analyzing the mRNA and protein levels by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively.

RESULTSThe cross-species dataset of HCC-related molecules included four genes: epidermal fatty acid-binding protein (E-FABP), liver (L)-FABP, tyrosine a-ketoglutarate transaminase (TKT), and cytokeratin (CK8). In humans, E-FABP mRNA expression was significantly higher (P less than 0.05) in HCC (0.87+/-0.14 vs. para-HCC: 0.64+/-0.12 and normal liver: 0.67+/-0.07; F=20.910). Similar results were obtained in tree shrew (HCC: 0.87 +/- 0.25 vs. para-HCC: 0.73 +/- 0.19 and normal liver: 0.68+/-0.19; F=3.807) and rat (HCC: 0.97+/-0.22 vs. para-HCC: 0.78+/-0.16 and normal liver: 0.80 +/- 0.13; F=4.482). The Western blotting analyses revealed a similar statistically significant trend.

CONCLUSIONThe cross-species screening strategy for tumor genes may represent a feasible and convenient process of identifying key molecule(s) for human HCC. E-FABP may be a particularly crucial molecule for hepatocarcinogenesis.

Adult ; Aged ; Animals ; Carcinoma, Hepatocellular ; metabolism ; Case-Control Studies ; Epidermis ; chemistry ; Fatty Acid-Binding Proteins ; metabolism ; Female ; Humans ; Liver ; metabolism ; Liver Neoplasms ; metabolism ; Male ; Middle Aged ; Rats ; Tupaiidae ; metabolism

Objective： The current study was designed to search into the detection of micrometastasis in peripheral blood in the patients with hepatocellular carcinoma(HCC)and its significance.Method： Peripheral blood samples were obtained from 65 patients with hepatocellular carcinoma, 21 non-HCC malignant tumors, 22 chronic hepatitis B or cirrhosis, and 21 cases of normal healthy volunteers. For identifying hepatocellular carcinoma cells in peripheral blood, the authors detected liver-specific alpha-fetoprotein(AFP) mRNA from total RNA extracted from whole blood by nested reverse transcription-polymerase chain reaction(Nested-RT-PCR). Results： AFP-mRNA was not detected in the normal healthy volunteers and the patient with non-HCC malignant tumors. The presence of AFP-mRNA in the patients with HCC (67.7％ ,44/65) was higher than those with chronic hepatitis B or cirrhosis ( 9.1％ ,2/22,P< 0.01). The detective rate of AFP-mRNA in the blood seemed to be correlated with the clinical stages of HCC, the presence of extrahepatic metastasis, and portal vein thrombosis. The positive AFP-mRNA in peripheral blood after treatment was correlated with the prognosis of the patients with HCC. AFP-mRNA was detected in 7 of 12 (58.3％) HCC patients with low level of serum AFP (< 25 μg/L). Conclusions： The presence of AFP-mRNA in peripheral blood may be an indicator of malignant or benign hepatocytes, which might predicate hematogenous spreading metastasis of tumor cells in the patients with HCC, it might be a valuable marker for predicting metastasis and recurrence of HCC, and it might play a supplementary role in the diagnosis of HCC with negative or low level of serum AFP.

OBJECTIVETo determine the methods for establishing an in vivo model of long-term hepatitis B virus (HBV) infection in the Chinese tree shrew (Tupaia belangeri chinensis).

METHODSSeventy-seven neonate (1-3 days old) and 49 young adult (2 weeks to 1 year old) tree shrews were inoculated with different HBV sources (chronic hepatitis B (CHB) human patient serum, single or pooled; HBV-infected tree shrew serum, single only; HBV-infected HepG2.2.15 cells' culture medium supernatant; HBV genome-transfected HepG2.2.15 cells' culture medium supernatant) through various routes of injection (subcutaneous, intraperitoneal, and direct liver via abdominal skin; adults also received intravenous and indirect liver via spleen). Serum and liver biopsies were collected from the animals at various time points post-inoculation for detection of HBV markers by fluorescence quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, time-resolved immunofluorescence, Southern blotting, dot blotting, immunohistochemistry, and microscopy.

RESULTSAmong the neonatal group of tree shrews, six (7.8%) were confirmed as HBV-infected for more than 72 (up to 228) weeks after inoculation and another seven (9.1%) were suspected of persistent infections. None of the young adult tree shrews developed persistent infection. Inoculation with single-source serum from either CHB humans or tree shrews were responsible for the most cases of infections, and the subcutaneous injection produced more infections than the other inoculation routes. The most reliable methods of determining HBV infection status were detection of serum HBV immunoreactive markers and intrahepatic HBV DNA.

CONCLUSIONIn order to establish an in vivo model of CHB in the tree shrew, the animals should be inoculated in the neonatal period using subcutaneous injection.

Animals ; Disease Models, Animal ; Female ; Hep G2 Cells ; Hepatitis B virus ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Tupaia

OBJECTIVETo observe the hepatitis B virus (HBV) replication in the tree shrews that were inoculated with HBV at neonatal period.

METHODSSix new-born tree shrews were inoculated with human HBV positive serum. Blood samples and liver biopsies were collected at different time points after inoculation. The HBV infection markers were tested by nested polymerase chain reaction (nPCR), fluorescence quantitative polymerase chain reaction (FQ-PCR), Southern blot, ELISA and immunohistochemistry staining. The liver tissues were observed under electron and light microscope.

RESULTS48 weeks after inoculation, HBV DNA and HBV cccDNA were detected in the serum and liver samples of three animals (number 1, 2 and 6) by nPCR. The copy-numbers of HBV DNA detected by FQ-PCR in their serum and liver samples were 103 and-104/ml respectively,and the total DNA in 1microg liver tissue was 107-108. Southern blot indicated that HBV replication intermediates such as HBV cccDNA and HBV ssDNA was detectable in liver tissues. HBsAg was detected by ELISA, and immunohistochemical staining showed a gradual increase of HBsAg-positive liver cells. High copy number of HBV DNA and suspected HBV EM particles could be detected in the liver samples from one of the three animals that have survived more than 2 years after inoculation. The other three animals showed low HBV DNA copy number, and the rest of the signs of HBV infection were negative or transiently positive.

CONCLUSIONSNeonatal tree shrews can be infected with human HBV. HBV can replicate inside the liver cells of tree shrew.

Animals ; Animals, Newborn ; Biopsy ; DNA, Circular ; analysis ; blood ; DNA, Viral ; analysis ; blood ; Disease Models, Animal ; Hepatitis B ; etiology ; pathology ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; physiology ; Humans ; Immunohistochemistry ; Liver ; pathology ; virology ; Polymerase Chain Reaction ; methods ; Tupaiidae ; Virus Replication

OBJECTIVEThis study was to clarify E-cadherin expressions in non-small cell lung cancer (NSCLC) and its correlation with patients' prognosis.

METHODSTissue microarrays (TMAs) containing specimens from 365 different NSCLC were constructed, covering all stages and almost all histological types of this disease. Slides were immunohistochemically stained with antibodies against E-cadherin. Expression pattern of the protein was analyzed with relation to the clinicopathological. Correlations of the results with patients' overall survival were also examined.

RESULTSImmunohistochemical staining revealed that E-cadherin protein was localized mainly on membranes and the cytoplasm of NSCLC tumors cells. Reduced E-cadherin expression was evident in 32.1%. Reduced E-cadherin expression significantly correlated with lymph nodes metastasis (chi(2) = 16.430, P = 0.001), histological dedifferentiation (chi(2) = 9.243, P = 0.010) and advanced clinical stage (chi(2) = 9.421, P = 0.024). There was no significant difference in E-cadherin expression between squamous cell carcinoma and adenocarcinoma. E-cadherin reduced expression correlated with a poor prognosis (P < 0.0001) in univariate analysis. Multivariate analysis showed a significantly lower survival probability for patients with reduced E-cadherin (P < 0.001), and E-cadherin was an independent prognostic factor for survival of NSCLC patients.

CONCLUSIONSIt suggests that dysfunction of E-cadherin has an important impact in the progression of lung cancer. As an independent prognostic factor, expression of E-cadherin can predict outcome of different group, together with conventional prognostic factors, and subsequently make appropriate management.

Adult ; Aged ; Aged, 80 and over ; Cadherins ; biosynthesis ; Carcinoma, Non-Small-Cell Lung ; metabolism ; mortality ; secondary ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; mortality ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Survival Rate

OBJECTIVETo screen the differentially expressed proteins especially at the precancerous stage of diethylnitrosamine (DEN) induced hepatocarcinogenesis by comparative proteome research.

METHODSRats were divided into normal and DEN groups and sacrificed periodically. The liver samples were stained with gamma-glutamyl transpeptidase (GGT) and HE to distinguish the preneoplastic lesion (pre-HCC) from the normal and HCC tissues. The two-dimensional electrophoresis (2-DE) and mass spectrometry (MALDI-TOF-MS/MS) were then applied to analyze the differentially expressed protein between pre-HCC and normal tissues, pre-HCC and HCC, as well as HCC and normal tissues. A few of the candidate proteins such as laminin receptor 1 (67LR) and agmatinase were validated by Western blot and RT-PCR.

RESULTSTotally, there were 82 proteins that differentially expressed two fold or more in one kind of tissues sample than the other, 47 of which occurred in the pre-HCC tissues. Eight proteins including 67LR were consistently up-regulated from normal tissue to pre-HCC and then to HCC tissues, while 22 proteins including agmatinase showed progressively down-regulated in these tissues samples.

CONCLUSIONThe protein expression profiles are different during the process of hepatocarcinogenesis. Further study on the differentially expressed protein, especially these upregulated in the precancerous stage such as 67LR and agmatinase, might contribute to prevention and early diagnosis of human HCC.

Animals ; Blotting, Western ; Carcinoma, Hepatocellular ; chemically induced ; metabolism ; pathology ; Diethylnitrosamine ; Liver ; metabolism ; pathology ; Liver Neoplasms, Experimental ; chemically induced ; metabolism ; pathology ; Male ; Neoplasm Proteins ; metabolism ; Precancerous Conditions ; metabolism ; pathology ; Proteins ; metabolism ; Proteome ; Rats ; Rats, Wistar ; Receptors, Laminin ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Ureohydrolases ; metabolism ; gamma-Glutamyltransferase

OBJECTIVETo detect the expression and variation of p53 gene during tree shrews' hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1).

METHODSTree shrews were divided into four groups: the tree shrews were infected with HBV and fed with AFB1 in group A, only infected with HBV in group B, fed with AFB1 alone in group C, and normal control in group D. All the tree shrews were performed liver biopsy every 15 weeks. The tissues of liver and tumor were detected by immunohistochemistry and molecular biotechnologies.

RESULTS(1) The incidence of hepatocellular carcinoma (HCC) in group A (66.7%) was higher than that in Group B and C (30%). HCC appearance in group A was earlier than that in group C (120.0 weeks +/-16.6 weeks vs 153.3 weeks +/-5.8 weeks, t = 3.336, P<0.01). (2) Mutated p53 protein was not found before the 75th week of the experiment in each group. (3) At the 105th week, the expression rates of mutated p53 protein were 78.6%, 60% and 71.4% in group A, B and C respectively, which were much higher than that (10%) in group D (x2 > or = 5.03, P<0.05). An abnormal band of p53 gene was detected in both group A and C. (4) The mutation points of p53 gene in liver cancer of tree shrew were at codon 275, 78 and 13. The nucleotide sequence and amino acids sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homology with those of human p53 respectively.

CONCLUSIONSThere is a remarkable synergistic effect between HBV and AFB1 on HCC. Mutated p53 protein is expressed before HCC occurrence, which promotes the development and progress of HCC. HBV and AFB1 may synergistically induce p53 gene mutation.

Aflatoxin B1 ; toxicity ; Animals ; Carcinoma, Hepatocellular ; genetics ; Cocarcinogenesis ; Gene Expression Regulation, Neoplastic ; Genetic Variation ; Hepatitis B ; virology ; Hepatitis B virus ; Liver Neoplasms, Experimental ; genetics ; Point Mutation ; RNA, Neoplasm ; analysis ; Tumor Suppressor Protein p53 ; genetics ; Tupaiidae