Objoctive:To investigate the expression of transforming growth factor-β(TGF-β)isoforms in colonic mucosa of ulcerative colitis(UC),and its role in the incidence of UC thereof.Methods:The expression of TGF-β isoforms was detected in colonic mucosa by immunohistochemical Envision method in UC patients,and the patients with irritable bowel syndrome (IBS)were used as controls.The relationship was analyzed between the expression of TGF-β isoforms and the degree of severity and extent of the disease.Results:There were statistical differences in the expression of TGF-β1 and TGF-β2 between the active stage UC,remission stage UC and IBS groups(P ＜ 0.01).The expression was significantly higher in active stage UC group than that in remission stage UC and 1BS groups(P＜ 0.01).The expression was significantly higher in remission stage UC group than that in IBS group(P ＜ 0.01).However,there was no statistical difference in the expression of TGF-β3 between active stage UC,remission stage UC and IBS groups(P＞ 0.05).The expression of TGF-β1 and TGF-β2 had positive correlation with the degree of sevefity(P ＜ 0.05).However,the expression of TGF-β3 had no linear correlation with the degree of severity(P ＞ 0.05).There was no linear correlation on the expressions of TGF-β1,TGF-β2 and TGF-β3 with extent of the disease(P ＞ 0.05).The degree of severity had positive correlation with extent of the disease(P ＜ 0.05).Conclusion:The expressions of TGF-β1 and TGF-β2 were enhanced in colonic mucosa of UC,and were correlated with the pathogenesis of UC.The expressions of TGFβ1 and TGF-β2 may serve as markers for assessing disease activity of UC.

Aged ; Giant Cells ; pathology ; Humans ; Male ; Stomach Neoplasms ; pathology

Objective To prepare hepatocyte growth factor(HGF) recombinant protein and confirm its activity preliminarily according to building HGF gene prokaryotic expression vector and transforming into E.coli.Methods Clone HGF inserted into the vector pET-26b(+) to construct prokaryotic expression vector pET-26b(+)-HGF and transform into E.coli Rosseta(DE3).The transformed bacteria induced by IPTG was purified through Ni-NTA resin affinity chromatography frozen-drying after renaturation.Results HGF gene recombinant prokaryotic expression vector pET-26b(+)-HGF was constructed successfully.E.coli Rosseta(DE3) which was transformed into pET-26b(+)-HGF expresses the target protein as the form of inclusion bodies,accounting for 38％ of the total bacterial proteins,and confirmed by Western blot.HGF protein which was purified by Ni-NTA resin affinity chromatography,has a purity of about 95％,and can promote proliferation,migration,and inhibition of apoptosis for human non-small cell lung cancer cell line A549 cells after interaction.Conclusion HGF gene recombinant prokaryotic expression vector pET-26b (+)-HGF was constructed and expressed in transformed E.coli Rosseta(DE3) successfully.They resumed their recombinant HGF protein structure after purification and renaturation,and had biological activity confirmed by in vitro studies.

AIMTo study the antitumor effect of baicalein on human leukemia K562 cell and its mechanism.

METHODSThe IC50 value and cytotoxity of K562 cell were detected by MTT method. The apoptotic cell was analyzed by FCM using Annexin V FITC--PI staining method. Sub-G1 peak was also measured by FCM. Protein expressions of Bcl-2, Fas, Caspase 3 were evaluated with FCM.

RESULTSBaicalein was shown to significantly inhibit the proliferation of K562 cell in a dose-dependent manner and selectively induce apoptosis of human leukemia K562 cells. Flow cytometric analysis showed that baicalein arrested K562 cells in the S phase. In addition, protein expression of Fas, Caspase 3 of K562 cells increased after exposure to baicalein, but Bcl-2 was unchanged.

CONCLUSIONBaicalein can selectively induce apoptosis of human leukemia K562 cell dose and time dependently through up-regulation of caspase-3 and fas gene expression level.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; Caspases ; metabolism ; Cell Cycle ; drug effects ; Flavanones ; Flavonoids ; pharmacology ; Humans ; K562 Cells ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Up-Regulation

This study is to investigate the influence and mechanism of action of asymmetrical dimethylarginine (ADMA) and the induced oxidative stress level on Alzheimer's disease (AD) incidence. ADMA concentration, nitric oxide, Abeta(40)/Abeta(42) ratio, inducible NO synthase (iNOS) activity and the concentrations of the induced free radicals including malondialdehyde (MDA), 3-nitrotyrosine (3-NT) and peroxynitrite (ONOO-) in the cerebrospinal fluid (CSF) from 34 neurologically normal controls and 37 AD patients were quantitatively determined and statistically compared. The results showed that the ADMA concentration significantly decreased in AD patients, and it showed negative correlation with the NO, iNOS activity, and showed positive correlation with MMSE score. ADMA concentration was negatively correlated with Abeta(40)/Abeta(42) ratio (P<0.01) with the observation that Abeta(40)/Abeta(42) ratio increased while ADMA level decreased in CSF in AD patients. The concentration levels of MDA, 3-NT and ROS significantly increased compared with the control with all the P values less than 0.05. These findings suggested that the ADMA disorder and the oxidative damage effect of the induced free radicals in CSF of AD patients are an important mechanism of AD incidence, and their joint regulation may provide new idea for the prevention and clinical treatment of AD.
Aged ; Alzheimer Disease ; cerebrospinal fluid ; Amyloid beta-Peptides ; cerebrospinal fluid ; Arginine ; analogs & derivatives ; cerebrospinal fluid ; Female ; Humans ; Male ; Malondialdehyde ; cerebrospinal fluid ; Middle Aged ; Nitric Oxide ; cerebrospinal fluid ; Nitric Oxide Synthase Type II ; cerebrospinal fluid ; Oxidative Stress ; Peptide Fragments ; cerebrospinal fluid ; Peroxynitrous Acid ; cerebrospinal fluid ; Reactive Oxygen Species ; cerebrospinal fluid ; Tyrosine ; analogs & derivatives ; cerebrospinal fluid

OBJECTIVETo evaluate the feasibility and efficacy of Narcotrend (NT) monitor in monitoring the depth of anesthesia in severely burned patients with target-controlled infusion (TCI) of remifentanil hydrochloride and propofol during perioperative period.

METHODSEighty patients with severe burn hospitalized from February to November 2011, to whom eschar excision was performed within one week after injury, were enrolled. They were classified into II to III grade according to the American Society of Anesthetists classification, and their total burn area ranged from 31% to 50%TBSA, or full-thickness burn area from 11% to 20% TBSA. Patients were divided into trial group (monitoring depth of anesthesia with routine method and NT monitor) and control group (monitoring depth of anesthesia with routine method) according to the random number table, with 40 cases in each group. All patients received TCI of remifentanil hydrochloride and propofol to induce and maintain anesthesia. During the operation, the anesthesia level of NT monitor used in the trial group was maintained from grade D1 to E0, while the fluctuation of mean arterial pressure (MAP) and heart rate of patients in control group was maintained around the basic values within a range of 20%, and on the basis of which, concentrations of two narcotics were adjusted. Concentrations of remifentanil hydrochloride and propofol during maintenance of anesthesia were recorded. The duration from drug withdrawal to waking from anesthesia (including the duration from drug withdrawal to eye opening by calling and the duration from drug withdrawal to orientation recovery) of patients was recorded. Values of MAP and heart rate at admission into the operation room, loss of consciousness, 2 min after intubation, before operation, 2, 15, and 30 min after the beginning of operation, and the end of operation were recorded. The prediction probability (P(k)) of NT stage (NTS) and NT index (NTI) in trial group, and that of MAP and heart rate in control group for two durations from drug withdrawal to waking form anesthesia were recorded. The administration of vasoactive drugs and intraoperative awareness of patients in two groups were recorded. Data were processed with t test, analysis of variance, and chi-square test, and the relationship between NTS, NTI, MAP, heart rate and their corresponding P(k) for the duration from drug withdrawal to orientation recovery was processed with Spearman correlation analysis.

RESULTSMaintained target effect-site concentration of remifentanil hydrochloride and target plasma concentration of propofol of patients were obviously lower in trial group [(2.62 ± 0.35) ng/mL, (3.84 ± 0.22) µg/mL] than in control group [(2.95 ± 0.21) ng/mL, (4.16 ± 0.31) µg/mL, with t values respectively -5.113 and -5.324, P values all below 0.01]. The duration from drug withdrawal to eye opening by calling and the duration from drug withdrawal to orientation recovery were obviously shorter in trial group [(10.2 ± 0.7) min, (11.1 ± 1.0) min] than in control group [(11.3 ± 1.0) min, (13.1 ± 0.7) min, with t values respectively -5.740 and -10.806, P values all below 0.01]. The MAP (except for 2 min after intubation) and the heart rate of patients in both groups were lower at the time points from loss of consciousness to the end of operation than at the time of entering operation room (with F values respectively 12.074, 36.425, P values all below 0.01 in trial group and F values respectively 21.776, 35.759, P values all below 0.01 in control group). The statistically significant difference between two groups in MAP level was only observed at the time of loss of consciousness (t = 3.985, P < 0.01). MAP level was close in two groups at other time points. Heart rates of patients in two groups were close during perioperative period. P(k) values of NTS and NTI for the duration from drug withdrawal to eye opening by calling (0.937 ± 0.025, 0.899 ± 0.049) were obviously higher than those of MAP and heart rate for this duration (0.579 ± 0.057, 0.536 ± 0.039, F = 900.337, P < 0.01). P(k) values of NTS and NTI for the duration from drug withdrawal to the orientation recovery (0.901 ± 0.031, 0.868 ± 0.046) were significantly higher than those of MAP and heart rate for this duration (0.532 ± 0.060, 0.483 ± 0.044, F = 890.895, P < 0.01). NTS, NTI, MAP, and heart rate were respectively negative, positive, positive and positive in correlation with their P(k) values for the duration from drug withdrawal to the orientation recovery (with r values from -0.734 to 0.682, P values all below 0.01). There was no statistically significant difference between two groups in administration of vasoactive drugs. No intraoperative awareness occurred.

CONCLUSIONSApplication of Narcotrend monitor in monitoring the depth of anesthesia in severely burned patients during perioperative period with TCI of remifentanil hydrochloride and propofol is beneficial to reducing dosage of narcotics and shortening duration of recovery from anesthesia, and it can accurately predict the level of consciousness of patients at the time of withdrawal of anesthesia.

Adolescent ; Adult ; Aged ; Anesthesia, Intravenous ; Burns ; surgery ; Female ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; instrumentation ; methods ; Piperidines ; Propofol ; Skin Transplantation ; methods ; Young Adult

BACKGROUNDCancer-testis antigen (CTA) is a family of the most noticeable tumor antigens which could be potential tumor markers for cancer diagnosis. In this research we aimed to investigate the expression of SSX-1 and NY-ESO-1 mRNA, two members of the CTA family, in tissue and peripheral blood of patients with hepatocellular carcinoma (HCC) to assess their feasibility for the immunotherapy and diagnosis of HCC and the association of their expression levels with diverse clinical indicators.

METHODSThirty-six north Chinese patients with HCC and 30 normal controls were enrolled in this study. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of SSX-1 and NY-ESO-1 mRNA in tumor tissues and corresponding levels in peripheral blood of patients.

RESULTSThe positive rates of SSX-1 and NY-ESO-1 mRNA expression were 61.1% (22/36) and 11.1% (4/36), respectively, in cancer tissues; 38.9% (14/36) and 5.6% (2/36), respectively, in the corresponding peripheral blood samples. No positive expression of either SSX-1 or NY-ESO-1 mRNA was detected in the samples of cancer-adjacent tissues, cirrhotic tissues, normal liver tissue or the peripheral blood of control patients. No significant relationship was found between the expression of these two genes and clinical indicators such as age, gender, tumor size, extent of differentiation, serum a-fetoprotein (AFP) level or infection with hepatitis B virus (P > 0.05). The short term recurrence rate was 46.2% (6/13) in patients whose peripheral blood expressed SSX-1 mRNA, while the recurrence rate in patients with negative SSX-1 mRNA was 28.6% (4/14).

CONCLUSIONSSSX-1 and NY-ESO-1 antigens might be new potentially promising targets for antigen-specific immunotherapy for HCC. High specific expression of SSX-1 and NY-ESO-1 mRNA suggested that we could apply them as tumor markers. The short term recurrence rate was significantly higher in patients whose peripheral blood expressed SSX-1 mRNA, suggesting that SSX-1 mRNA could be used as indicator for recurrence, metastasis and prognosis of HCC.

Adolescent ; Adult ; Aged ; Antigens, Neoplasm ; genetics ; Carcinoma, Hepatocellular ; metabolism ; Female ; Humans ; Liver Neoplasms ; metabolism ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Neoplasm Proteins ; genetics ; Neoplastic Cells, Circulating ; RNA, Messenger ; analysis ; blood ; Repressor Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the occurrence, management and prognosis of fatal pulmonary embolism in patients underwent coronary intervention in our department.

METHODSeven patients had fatal pulmonary embolism after coronary intervention in six years, we analysis each patient by the occurrence, prognosis, management of the disease.

RESULTSDuring the last 6 years, 7 [five males, mean age (55.9 +/- 11.7) years, 5 after coronary angiography and 2 after percutaneous coronary intervention] patients developed fatal pulmonary embolism after PCI. All 7 patients presented respiratory and cardiac arrest within 24 hours post coronary intervention. Three patients died, one patient experienced brain death and another three patients survived and are alive without complication till now.

CONCLUSIONThe fatal pulmonary embolism is a scarce complication after coronary intervention with high acute mortality and satisfactory outcome for survivors.

Adult ; Aged ; Angioplasty, Balloon, Coronary ; adverse effects ; Humans ; Male ; Middle Aged ; Prognosis ; Pulmonary Embolism ; diagnosis ; etiology ; Treatment Outcome

OBJECTIVETo modify the current PCR-based method for rapid and efficient preparation of small interfering RNA (siRNA) expression cassette to improve the efficiency of RNA interference.

METHODSThe U6 promoter sequence was amplified by PCR using the genomic DNA of K562 cells as the template, and cloned into pMD18-T vector which served as the template for further PCR amplification with the primers on the plasmid. The amplified product was directly used as the template for preparing siRNA expression cassette. The siRNA expression cassette targeting p53 gene was amplified, verified by sequencing, and transfected into SH-SY5Y cells. After a 48-hour transfection, the cells were harvested and the total RNA was for RT-PCR for evaluating the effect of RNA interference.

RESULTSThe sequencing result confirmed the correct U6 promoter sequence cloned from K562 cells. After transfection of SH-SY5Y cells for 48 h with siRNA expression cassette, the p53 gene expression was inhibited at the mRNA level in comparison with the control cells as demonstrated by RT-PCR detection.

CONCLUSIONThe siRNA expression cassette prepared using the established method described hereby can be well applicable in RNA interference research.

Gene Silencing ; Gene Targeting ; methods ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Polymerase Chain Reaction ; RNA, Small Interfering ; biosynthesis ; genetics ; RNA, Small Nuclear ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVETo study gene expression of collagen types IX and X in human lumbar intervertebral discs during aging and degeneration and to explore the role of collagen types IX and X in disc degeneration.

METHODSFetal, adult and pathologic specimens were subjected to in situ hybridization with cDNA probes to investigate mRNA-expressions of types IX and X collagen gene.

RESULTSIn fetal intervertebral discs, positive mRNA hybridization signals of type IX collagen were concentrated in the nucleus pulposus and the inner layer of anulus fibrosus. Interstitial matrix of the nucleus pulposus also showed positive type X collagen staining. Positive mRNA hybridization signals of types IX and X were not detected in the middle and outer layers of anulus fibrosus. In adult specimens, expression of type IX collagen mRNA was markedly decreased. No hybridization signals of type X collagen was observed. As for pathological specimens, there was no gene expression of type IX collagen. In severe degenerated discs from adults, there were focal positive expressions of type X collagen.

CONCLUSIONSObvious changes of collagen gene expression occur with aging. Expression of type IX collagen decreases in adult and pathological discs. Results of type X collagen expression suggest that type X collagen is expressed only in older adult and senile discs (i.e., when disc degeneration has already reached a terminal stage), indicating the terminal stage of degeneration.

Adolescent ; Adult ; Collagen Type IX ; metabolism ; Collagen Type X ; metabolism ; Female ; Gene Expression ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Intervertebral Disc ; embryology ; metabolism ; Lumbar Vertebrae ; Male