Animals ; Gene Expression ; Hypoxia ; metabolism ; Prefrontal Cortex ; metabolism ; Rats ; Somatomedins ; metabolism

Background and purpose: Ursolic acid is widely present in spica prunellae, hedyotis diffusa and other heat antidotes. The growth of a variety of tumor cells can be inhibited and induced apoptosis by ursolic acid.This study was aimed to investigate the effect and possible mechanisms of UA on inducing apoptosis of human gastric carcinoma BGC823 cells. Methods: The MTT assay was used to detect the antiproliferative effect of UA on BGC823 cells. Flow cytometry was used to detect cell cycle and apoptosis of BGC823 cells. The expression level of bcl-2 and bax gene was investigated by real time-polymerase chain reaction (real time-PCR). Results: UA inhibited the proliferation of BGC823 cells in a dose and time-dependent way. After treatment by UA for 24. 48 and 72 h, the IC_(50) of BGC823 was 36.88, 34.72, and 32.18 μmol/L, respectively. UA could signifcantly induce apoptosis of BGC823 cells and block cells at G_2/M phase. UA could increase the expression of bax gene and decrease the expression of bcl-2 gene in a dose and time-dependent way. Conclusion: UA could induce apoptosis and inhibit the proliferation of BGC823 cells in a dose and time-dependent way. It could arrest cell cycle of BGC823 cells at G_2/M phase. Its mechanisms might be associated with the up-regulation of bax gene and down-regulation of bcl-2 gene.

Objective To investigate the risk factors of severe hemolysis during extracorporeal membrane oxygenation (ECMO). Methods The clinical data of adult patients undergoing ECMO after cardiac surgery admitted to Fuwai Hospital from December 2010 to October 2015 were retrospectively analyzed. Demographic characteristics, renal function, primary disease, operation data, ECMO related data and outcomes were recorded. Patients were divided into normal free hemoglobin (FHB) group (FHB ≤ 500 mg/L) and severe hemolysis group (FHB > 500 mg/L) according to the FHB level during ECMO support. The parameters before and after ECMO support were compared between the two groups. Logistic regression was used to identify the independent risk factors of severe hemolysis. Results A total of 81 patients including 19 patients with severe hemolysis was enrolled, and 62 in normal FHB group. There was no difference in cardiopulmonary bypass (CPB) time, clamping time, lactate level before ECMO, cardiopulmonary resuscitation, intra-aortic balloon pump use and central catheter insertion between two groups. The maximums of serum creatinine (SCr) and FHB levels were higher in severe hemolysis group as compared with those in normal FHB group [maximal SCr (μmol/L): 281.02±164.11 vs. 196.67±87.31, maximal FHB (mg/L): 600 (600, 700) vs. 200 (100, 300)], the incidence of clots in circuit or oxygenator, infection, and hemofiltration in severe hemolysis group was increased [26.3% (5/19) vs. 4.8% (3/62), 31.6% (6/19) vs. 12.9% (8/62), 36.8% (7/19) vs. 14.5% (9/62), all P < 0.1]. As well as outcomes including the rate of site of surgery or intubation bleeding and acute renal failure [ARF, 57.9 % (11/19) vs. 30.6% (19/62), 94.7% (18/19) vs. 41.9% (26/62)], and the survival rate was lowered [10.5% (2/19) vs. 51.6% (32/62), all P < 0.05]. As result of univariate analysis, clots in circuit or oxygenator, infection and hemofiltration were associated with severe hemolysis. It was showed by logistic regression analysis that the clots in circuit or oxygenator was a risk factor of severe hemolysis during ECMO [odds ratio (OR) = 6.262, 95% confidence interval (95%CI) = 1.244-31.515, P = 0.026]. Conclusions The clots in circuit or oxygenator were independent risk factors of severe hemolysis during ECMO. Severe hemolysis can induce the increase of the rate of bleeding in the operation site or intubation and the rate of ARF, and decrease of the survival rate.

OBJECTIVE: Studying forensic medical problem related with RTA leading PTSD and supplying accumulating evidence for psychiatric compensation in court. METHODS: One hundred and fifty six victims of RTA were recruited who applied to court for a costs order. The victims were examined for psychiatric diagnosis by psychiatrists and for rank of impairment by experts in forensic clinical medicine. The self-report psychopathological status and quality of life were also measured. RESULTS: Eighty one victims of 156 (51.92%) fulfilled the criteria for PTSD (ICD-10). Morbidity difference in male and female were significant; The more serious extent of impairment is, the more PTSD'possibility is; The scores in World Health Organization Quality of Life were lower and in SAS and SDS were higher in PTSD group than in non-PTSD group. Acquirement of awarded costs could obviously prevent PTSD. CONCLUSION The higher PTSD incidence existed in the RTA victims who applied to court for a costs order, and acquirement of awarded costs could obviously prevent PTSD.
Accidents, Traffic ; Adolescent ; Adult ; Compensation and Redress ; Disability Evaluation ; Female ; Follow-Up Studies ; Forensic Medicine ; Humans ; Male ; Middle Aged ; Psychiatric Status Rating Scales ; Quality of Life ; Stress Disorders, Post-Traumatic/psychology* ; Wounds and Injuries/psychology* ; Young Adult

OBJECTIVETo discuss the optimal immunization dose by observing the immunoprotective effects of different doses of recombinant Schistosoma japonicum (Chinese strain) signaling protein 14-3-3 (rSj14-3-3).

METHODSSj14-3-3 gene was amplified by reverse transcriptase PCR (RT-PCR), subcloned into prokaryotic expression vector pET28a, then transformed into E.coli to express by inducing. Purified rSj14-3-3 was prepared through SDS polyacrylamide gel electrophoresis (SDS-PAGE), electroelution, dialysis, then BALB/c mice were divided into 5 groups and immunized in rSj14-3-3 protein followed by challenging infection (the 1st, 2nd, and 3rd groups were immunized in 50 microg, 100 microg and 300 microg antigen, respectively. The 4th, 5th groups were immunized in Freund's adjuvant and normal saline controls). After 6 weeks of challenging infection, the mice were killed and the worm and egg reduction rates were calculated. And the mice sera in different time were taken to examine the specific anti-Sj14-3-3 IgG.

RESULTSrSj14-3-3 protein was expressed successfully. After immunizing and challenging, worm reduction was found to be 28.20% in the 1st group, 43.10% in the 2nd group, 40.00% in the 3rd group, respectively. Number of eggs in liver tissue was reduced by 41.80%, 57.50%, 55.70%, respectively. Compared the results of the tested groups to the controls, the differences were of significance by t-test (worm reduction rate: t = 6.8 in the 1st group, t = 8.7 in the 2nd group, t = 7.3 in the 3rd group, P < 0.01 in all tested groups. Egg reduction rate at the group's number above: t = 11.23, t = 11.54, t = 7.99, P < 0.01 in all tested groups). As compared the results between the tested groups by chi(2), the differences were of significance between the 1st and the 2nd groups (worm reduction rate: chi(2) = 8.96, P < 0.05; egg reduction rate: chi(2) = 15.69, P < 0.05), between the 1st and the 3rd groups, the differences were also of significance (worm reduction rate: chi(2) = 6.52, P < 0.05; egg reduction rate: chi(2) = 12.52, P < 0.05). The difference was not of significance between the 2nd and the 3rd groups (worm reduction rate: chi(2) = 1.20, P > 0.05; egg reduction rate: chi(2) = 0.93, P > 0.05). In all tested groups, total anti-Sj14-3-3 specific IgG rose markedly. IgG(1) and IgG(2a) subtypes were high, but IgG(2b) and IgG(3) were near the background in four subtypes tested.

CONCLUSIONImmunoprotection of rSj14-3-3 should have some relations with immunization dose, and the protection obtained from immunizing mice by using 100 microg antigen was the best.

14-3-3 Proteins ; administration & dosage ; immunology ; Animals ; Antibodies, Helminth ; immunology ; Antibody Formation ; Antigens, Helminth ; blood ; Female ; Helminth Proteins ; immunology ; Immunoglobulin G ; blood ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; Schistosoma japonicum ; genetics ; immunology ; Signal Transduction ; Vaccination

OBJECTIVETo clone and analyze duck hepatitis B virus genome from Chongqing brown duck.

METHODSDuck hepatitis B virus (DHBV) DNA extracted from a Chongqing brown duck was amplified by PCR and cloned into PGEM-T vector using T-A clone method. The sequence of this DHBV genome was analyzed with some softwares after identified.

RESULTSThe duck hepatitis B virus genome from Chongqing brown duck (DHBVcq), which was 3 024 nucleotides long, contained three ORFs whose onset and end nucleotides were in accord with those of HPUGA, encoding P, PreC/C and PreS/S protein respectively. Comparison of this strain with other DHBV reported in GenBank showed that the homology of DHBVcq and M32990 got the highest score of 94.9% at nucleotide level, while DHBVcq and DHBVCG got the least (89.8%). Most of the conserved regulation nucleotides and amino acids sequence found in other DHBV were also identified in DHBVcq. The epsilon region of DHBVcq, which was important for encapsidation of pgRNA and synthesis of minus-strand DNA, differed from that of most other DHBV strains, forming a stem-loop conformation with a three- nucleotides upper stem rather than a common nine-nucleotides one in free status.

CONCLUSIONThe successful clone and analysis of DHBVcq provide further studies with helpful information.

Animals ; Cloning, Molecular ; DNA, Viral ; chemistry ; genetics ; Ducks ; Hepatitis B Virus, Duck ; classification ; genetics ; Hepatitis Virus, Duck ; genetics ; Open Reading Frames ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology

Objective To observe the methylation and expression of 14-3-3 sigma gene in human breast cancer.Methods 40 breast cancer tissues and 18 mammary gland tissues with benign lesions were analyzed by methylation specific PCR(MSP),RT-PCR,and Western-blot(WB)so as to detect the methylation status and expression of 14-3-3 sigma mRNA or protein.Results Methylation of 14-3-3 sigma gene was detectable in 85%(34/40)of patients with breast cancers.RT-PCR showed negative in 12.5% of breast cancers(5/40),WB also indicated that 14-3-3 sigma was not detected in 32 of 40 breast carcinomas (80%).Furthermore,both RT-PCR and WB were negative in 30 of 34 positive cases by MSP.While methylation of 14-3-3 sigma was not detectable and its expression was demonstrated by RT-PCR and WB among 18 cases of benign breast diseases.These evidences proposed that methylation of 14-3-3 sigma gene had great relevance with its silence.Conclusion Methylation and loss of expression in 14-3-3 sigma gene were high frequent events in breast cancers.And methylation of 14-3-3 sigma gene might be related to its loss.

A rapid method of Liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) combined with pyridinium chlorochromate (PCC) oxidation has been developed to determine chemical structures of two novel isomers in bear bile powder. Derivatives of ursodeoxycholic acid (UDCA) and chenodeoxycholic acid (CDCA) were semi-synthesized by PCC oxidation, then were analyzed by LC-Q-TOF-MS. Separation was carried out on a reverse column with the mobile phase of acetonitrile-0.1% formic acid (45:55). The data of Q-TOF-MS was acquired by MS, MS/MS, positive and negative modes. Since UDCA and CDCA were stereochemical isomeric at an alcohol position, two oxidation products were same and have been confirmed by LC-Q-TOF-MS. Other two products were also determined based on the PCC oxidation theory. Samples of bear bile powder were dissolved by methanol and measured by LC-Q-TOF-MS. Two unknown peaks were found and identified by matching their retention times and accurate mass spectra ions with PCC oxidation productS. Finally, the structures of two new bile acids in bear bile powder were confirmed as 3alpha-hydroxy-7-oxo-5beta-cholanic acid, 7alpha-hydroxy-3-oxo-5beta-cholanic acid, respectively.
Animals ; Bile ; chemistry ; Chromatography, Liquid ; methods ; Isomerism ; Oxidation-Reduction ; Powders ; chemistry ; Pyridinium Compounds ; chemistry ; Tandem Mass Spectrometry ; methods ; Tissue Extracts ; chemistry ; Ursidae ; Ursodeoxycholic Acid ; chemistry

OBJECTIVETo study the purification, refolding and bioactivity of streptavidin-tagged human tumor necrosis factor-alpha (SA-TNF-alpha) bi-functional fusion protein.

METHODSSA-TNF-alpha fusion protein was expressed in BL21(DE3) host bacteria, purified using Ni-NTA affinity chromatography and refolded by dilution and dialysis followed by identification using Western blotting. The effect of SA-TNF-alpha fusion protein against L929 cells was evaluated by MTT assay. Flow cytometry was used to analyze the surface modification of biotinylated MB49 tumor cells by SA-TNF-alpha fusion protein.

RESULTSRecombinant SA- TNF-alpha fusion protein was expressed in BL21(DE3) at about 30% of total bacterial protein, with a purity of about 95% after purification. The SA-TNF-alpha fusion protein existed as dimmers, tetramers and higher order structures after refolding. The fusion protein exhibited a bi-functionality by inhibiting L929 cells and SA-mediated high-affinity binding to biotinylated cell surfaces, with an anchor modification rate of above 90%.

CONCLUSIONThe dimmers, tetramers and higher order structures of the obtained SA-TNF-alpha fusion protein all exhibit a bi-functionality, and may serve as a potential candidate therapeutic agent for tumors.

Chromatography, Affinity ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Nickel ; Protein Folding ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Streptavidin ; biosynthesis ; genetics ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

Adenosine Triphosphatases ; chemistry ; Antineoplastic Agents ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Drug Screening Assays, Antitumor ; Female ; Humans ; Liver Neoplasms ; pathology ; Luminescence ; Luminescent Measurements ; Male ; Middle Aged ; Tumor Cells, Cultured