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.

OBJECTIVETo clone the coding gene of the stage-specific antigen cC1 from Cysticercus cellulosae and express high levels of soluble cC1 in E.coli.

METHODSThe cC1 gene was amplified from Cysticercus cellulosae by RT-PCR and cloned into pMD18-T vector, followed by subcloning into the prokaryotic expression plasmid pET28a. The recombinant plasmid was transformed into E.coli BL21(DE3) and the expression conditions were optimized. The expressed product was purified by Ni(+)-affinity chromatography, analyzed by high-performance liquid chromatography (HPLC), and identified with SDS-PAGE and Western blotting.

RESULTSThe fragment length of the amplification product by RT-PCR was 1056 bp. Comparison of the amplified gene sequence with the cC1 gene in Genbank identified a samesense point mutation at 423 position in the gene cloned into the expression plasmids. After a 6-h induction with 0.05 mmol/L IPTG at 37 degrees celsius;, the expression of the 40 kd soluble fusion protein exceeded 60% of the total bacterial protein, and the fusion protein was recognized by Cysticercus-infected human sera. The purity of the fusion protein was about 94% after purification by affinity chromatography.

CONCLUSIONThe stage-specific antigen cC1 from Cysticercus cellulosae has been successfully cloned and the soluble protein efficiently expressed in E.coli, which provides the basis for its further study and application.

Animals ; Antigens, Helminth ; biosynthesis ; genetics ; immunology ; Cloning, Molecular ; Cysticercus ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Solubility ; Swine ; Taenia solium ; immunology

OBJECTIVETo study the effect of nerve growth factor (NGF) and Schwann cells on axon regeneration of the inferior alveolar nerve following mandibular lengthening with distraction osteogenesis.

METHODSUnilateral mandibular osteodistraction was performed in 9 healthy adult male goats with a distraction rate of 1 mm/d. Every 3 goats were killed on days 7, 14 and 28 after mandibular lengthening, respectively. The inferior alveolar nerves in the distraction callus were harvested and processed for ultrastructural and NGF immunohistochemical study. The inferior alveolar nerves from the contralateral side were used as controls.

RESULTSOn day 7 after distraction, axon degeneration and Schwann cell proliferation were observed, and very strong staining of NGF in the distracted nerve was detected. On day 14 after distraction, axon regeneration and remyelination were easily observed, and NGF expression started to decline. On day 28 after distraction, the gray scale of NGF immunoreactivity recovered to the normal value and the Schwann cells almost recovered to their normal state.

CONCLUSIONSGradual mandibular osteodistraction can result in mild or moderate axon degeneration of the inferior alveolar nerve. Nerve trauma may stimulate the proliferation of Schwann cells and promote the synthesis and secretion of NGF in the Schwann cells. Schwann cells and NGF might play important roles in axon regeneration of the injured inferior alveolar nerve following mandibular lengthening.

Animals ; Axons ; pathology ; physiology ; Goats ; Immunohistochemistry ; Male ; Mandible ; surgery ; Mandibular Nerve ; physiology ; Nerve Growth Factor ; physiology ; Nerve Regeneration ; physiology ; Osteogenesis, Distraction ; Schwann Cells ; physiology

OBJECTIVETo obtain streptavidin-tagged human interleukin-21 (hIL21) fusion protein and evaluate its bioactivities.

METHODShIL21-SA-pET21 and pET24a-SA- hIL21 plasmids were constructed and expressed in BL21(DE3) host bacteria. The hIL21-SA and SA- hIL21 fusion protein were purified through Ni-NTA affinity chromatography and refolded by dialysis. Flow cytometry was used to detect hIL21-SA and SA- hIL21 fusion protein on the biotinylated MB49 tumor cells. MTT assay was used to evaluate the effect of the fusion protein on the proliferation of human peripheral blood lymphocytes (PBLs) stimulated by Anti-CD3.

RESULTSThe recombinant fusion proteins were highly expressed in BL21(DE3) at about 30% of the total bacterial proteins. The two fusion proteins exhibited bifunctional activities, i.e. both biotin-binding property and hIL21 activity and SA-mediated high-affinity binding to biotinylated cell surfaces (with anchoring modified rate of about 95.18% and 96.91%).

CONCLUSIONWe have successfully obtained bifunctional fusion protein hIL21-SA and SA- hIL21,which will provide a basis for further study of tumor biotherapy using the proteins.

Cancer Vaccines ; immunology ; Cell Line, Tumor ; Escherichia coli ; genetics ; metabolism ; Humans ; Interleukins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Streptavidin ; biosynthesis ; genetics