This study investigated the changes of CD4(+) CD25(+) regulatory T cells (Tregs) in peripheral blood of patients with hepatocellular carcinoma before and after transcatheter arterial chemoembolization (TACE). The proportion of CD4(+) CD25(+) Tregs among CD4(+) T lymphocytes in peripheral blood of 33 patients with hepatocellular carcinoma was determined by flow cytometry before, 1 week and 1 month after TACE. And 25 healthy volunteers served as control. One month after TACE, the patients were divided into two groups: 22 in group A, who were in stable condition or getting better; and 10 in group B, who were deteriorating. One patient died and was excluded. The results showed that the percentage of CD4(+)CD25(+) Tregs among CD4(+) T lymphocytes did not significantly change in the 33 patients 1 week after TACE as compared with that before TACE, however, the difference was significant (P<0.01) between the patients with hepatocellular carcinoma and the healthy subjects. The percentage of CD4(+) CD25(+) Tregs among CD4(+) T lymphocytes in group A 1 month after TACE was decreased significantly in comparison with that before and 1 week after TACE (P<0.01), whereas, that in group B was increased significantly 1 month after TACE (P<0.01). It was concluded that patients with hepatocellular carcinoma had a higher proportion of CD4(+)CD25(+) Tregs in peripheral blood. TACE did not significantly affect the level of CD4(+) CD25(+) Tregs within short time (such as 1 week). The proportion of CD4(+)CD25(+) Tregs in peripheral blood 1 month after TACE was related to the prognosis of hepatocellular carcinoma.
Antineoplastic Agents/administration & dosage ; Carcinoma, Hepatocellular/*immunology ; Carcinoma, Hepatocellular/therapy ; Chemoembolization, Therapeutic/*methods ; Liver Neoplasms/*immunology ; Liver Neoplasms/*therapy ; T-Lymphocytes, Regulatory/*immunology

OBJECTIVETo review the clinical manifestations, imaging, tumor markers, treatment methods, pathology results and clinical curative effects of pineal region tumors and to evaluate the characteristics and intervention strategies for those tumors.

METHODSThe clinicopathological data of 132 patients with pineal region tumor treated in our department between January 2000 and May 2008 were retrospectively studied.

RESULTSA moderate predominance in males was presented. The clinical manifestations of the disease included increased intracranial pressure and ocular movement impairment. There were some features but no regularity and specific appearance on imaging including CT and MRI. 88.6% of patients associated with hydrocephalus. A high serum level of alpha-fetoprotein (AFP) was presented in 14 cases and high HCG in 9 cases. Eighteen cases received direct radiation therapy and 7 had radiotherapy post biopsy. 107 cases were treated surgically and 63 cases received postoperative adjuvant treatment. 114 cases had pathology results including 56 germ cell tumors. The patients were followed up for 12 approximately 132 months. Recurrence developed in 23 cases and 12 cases died. The 5-year survival rate was 89.3%.

CONCLUSIONPineal region tumors are often associated with hydrocephalus and this makes preoperative diagnosis difficult. Imaging examination may help diagnosis but less specific. Germ cell tumors may diagnosed by some tumor markers. Radiation therapy is the choice of treatment for pure germinomas. Other types of pineal region tumors should receive surgical treatment. Postoperative adjuvant treatment based on pathology can provide a good prognosis in pineal region tumor.

Adolescent ; Adult ; Aged ; Brain Neoplasms ; blood ; diagnosis ; therapy ; Child ; Child, Preschool ; Chorionic Gonadotropin ; blood ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Hydrocephalus ; etiology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Pineal Gland ; pathology ; surgery ; Pinealoma ; blood ; diagnosis ; therapy ; Retrospective Studies ; Sex Factors ; Survival Rate ; Tomography, X-Ray Computed ; Young Adult ; alpha-Fetoproteins ; metabolism

OBJECTIVETo establish transgenic mouse models of familial amyotrophic lateral sclerosis (FALS) and identify the genotype of the first filial generation.

METHODSSix male B6SJL SOD1G93A/+ hemizygote mice were mated with 6 female B6SJLF1/J+/+ mice to produce the filial generation. The genomic DNA was extracted from the tail vein blood of the first filial generation mice and PCR was performed to amplify the hmSOD1 gene fragment. The genotype of the mice was determined by electrophoresis, and the PCR product was purified for further gene sequence analysis and detection of mutation loci.

RESULTSFifty-three progeny mice were born and the survival rate before ALS onset was 98% (52/53), and among the survived mice, the positivity rate for hmSOD1 gene was 44.2% (23/52). Electrophoresis result showed that the PCR product of 236 bp was consistent with the hmSOD1 gene fragment, and the sequence of the PCR product was identical with hmSOD1 gene sequence of G93A mutant.

CONCLUSIONTransgenic mouse models of ALS can be established in the first filial generation of male B6SJL SOD1G93A/+ mice mated with female B6SJLF1/J+/+. PCR technique can precisely identify the genotype of the filial generation.

Amyotrophic Lateral Sclerosis ; enzymology ; genetics ; pathology ; Animals ; Animals, Newborn ; Base Sequence ; Breeding ; Disease Models, Animal ; Electrophoresis, Agar Gel ; Female ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Mice, Transgenic ; Point Mutation ; Sequence Analysis, DNA ; Superoxide Dismutase ; genetics ; Superoxide Dismutase-1

OBJECTIVETo develop an effective method for screening recombinant hairpin RNA expression plasmids using single restriction endonuclease analysis.

METHODSThe double-strand DNA fragment containing a ClaI site (the flanking sequences of which were not complementary) was annealed and ligated into small hairpin RNA (shRNA) expression vector pSilencer-4.1 that did not contain ClaI site to construct the circular pSilencer-4.1-ClaI vector. With BamHI and HindIII, the pSilencer-4.1-ClaIwas digested and ligated with the DNA template of green fluorescence protein (GFP) shRNA that did not include a ClaI site. The plasmid DNA of the positive clones was extracted and digested with ClaI, and the inserted DNA sequence of the non-linearized plasmid was identified by sequence analysis.

RESULT AND CONCLUSIONDNA sequencing showed that pSilencer-4.1-ClaI was correctly constructed and the plasmids resistant to ClaI digestion were all recombinant vectors encoding GFP shRNA. The constructed pSilencer-4.1-ClaI can be used as a universal vector to construct the shRNA expression plasmid, and the incorporated ClaI sites may allow efficient screening of recombinant shRNA expression vectors.

Base Sequence ; Gene Expression ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Inverted Repeat Sequences ; Molecular Sequence Data ; Plasmids ; genetics ; RNA, Small Interfering ; genetics ; Restriction Mapping ; methods ; Sequence Analysis, DNA ; Time Factors

OBJECTIVETo study the MRI manifestation of testicular tumor and the value of MRI in the diagnosis of the disease.

METHODSWe retrospectively analyzed 23 cases of pathologically confirmed testicular tumor, and observed the morphological characteristics, signals and surrounding conditions of the tumor using plain and enhanced MRI scanning.

RESULTSOf the 23 cases, seminoma was identified in 7, mixed germinoma in 3, teratoma in 3, endodermal sinus tumor in 2, epidermoid in 1, Leydig cell tumor in 1, leucoma in 1, nonspecific inflammatory mass in 3, and tuberculosis in 2. MRI revealed the precise locations and specific characteristics of

CONCLUSIONBased on MRI findings and clinical manifestation, most testicular tumors can be diagnosed correctly.

Adolescent ; Adult ; Carcinoma, Embryonal ; diagnosis ; Child ; Child, Preschool ; Endodermal Sinus Tumor ; diagnosis ; Germinoma ; Humans ; Infant ; Leydig Cell Tumor ; diagnosis ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Seminoma ; diagnosis ; Teratoma ; diagnosis ; Testicular Neoplasms ; diagnosis ; Young Adult

Objective To compare the efficacy of transplanting bone marrow mesenchymal stem cell (BMSC) or microenvironmental induced BMSC ( iBMSC) into the ischemic myocardium of rats with myocardial infarction. Methods iBMSC was defined as BMSC co-cultured with myocardial cells for 2 weeks. The stem cells or equal volume PBS were injected into ischemic border zone 1 wk after experimental infarction. Cardiac performance was evaluated at 1, 2, and 4 wk after cell transplantation by echocardiography and analyzed histologically at 4 wk after cell transplantations. Results Compared with PBS group, both BMSC and iBMSC transplantations reduced infarct size. iBMSC enhanced the beneficial effects of BMSC on improving cardiac function (FS: 28.5% ±4.3% in PBS, 29.0% ±2.0% in BMSC and 45. 1% ±3. 1% in iBMSC group at 4 weeks post transplantation, iBMSC group vs. PBS group P <0. 05, iBMSC group vs. BMSC group P <0. 05). Immunofluorescence microscopy results revealed co-localization of SPIO-labeled transplanted cells with cardiac markers for cardiomyocytes, indicating regeneration of damaged myocardium. Conclusion Our data suggest that iBMSC implantation is more effective on improving cardiac function than BMSC implantation in this model. iBMSC might serve as a new promising therapeutic cell source for regenerating ischemic myocardium in patients with post-infarction heart failure.

OBJECTIVETo construct an eukaryotic expression vector for miRNA-1-2 that can be expressed in rat H9C2 cardiomyocytes.

METHODSThe precursor miRNA (pre-miRNA) DNA template for miRNA-1-2 was designed and generated by PCR amplification. The DNA template was inserted into the hairpin RNA expression vector pSilence-4.1-neo and identified by DNA sequencing analysis. The recombinant plasmid DNA was then transfected into H9C2 cells via Lipofectamine, and the green fluorescence protein expression vector pEGFP-N3 served as the transfection marker. Twenty-four hours after transfection, the total cellular RNA was extracted using TRIzol reagent, and thermoscript reverse transcriptase (RT)-PCR was performed to determine miRNA-1-2 precursor expression.

RESULTS AND CONCLUSIONDNA sequencing indicated that the miR-1-2 expression plasmid was correctly constructed. The precursor miRNA-1-2 was successfully expressed in the H9C2 cells, and the expression of Hand2 protein could be efficiently inhibited by miRNA-1.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Line ; Down-Regulation ; Green Fluorescent Proteins ; genetics ; metabolism ; MicroRNAs ; genetics ; Myocytes, Cardiac ; cytology ; metabolism ; Plasmids ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Rats ; Transfection

The effects of ketamine on transient outward potassium current (I(to)) of isolated human atrial myocytes were investigated to understand the mechanism of part of its effects by whole-cell patch-clamp. Atrial myocytes were enzymatically isolated from specimens of human atrial appendage obtained from patients under going cardiac valve displacing. Ito is recorded in voltage-clamp modes using the patch-clamp technique at room temperature. Currents signals were recorded by an Axopatch 200B amplifier with the Digidata 1322A-pClamp 9.0 data acquisition system. Ketamine decreased I(to) of human atrial myocytes in a dose-dependent manner. The current-voltage curve was significantly lowered, 30, 100, 300, and 1000 micromol x L(-1) ketamine decreased respectively I(to) current density about (13.62 +/- 0.04)%, (38.92 +/- 0.05)%, (72.24 +/- 0.10)% and (83.84 +/- 0.05)% at the potential of 50 mV, with an IC50 of 121 micromol x L(-1). The I(to) activation curve, inactivation curve and the recovery curve were not altered by ketamine. So, ketamine concentration-dependently decreased I(to) of human atrial myocytes.
Adolescent ; Adult ; Aged ; Anesthetics, Dissociative ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Heart Atria ; cytology ; Humans ; Ketamine ; administration & dosage ; pharmacology ; Male ; Middle Aged ; Myocytes, Cardiac ; cytology ; drug effects ; physiology ; Patch-Clamp Techniques ; Potassium Channels ; drug effects ; Young Adult

OBJECTIVETo study the effect of high glucose on GRK2 gene expression in H9C2 cardiomyoblasts in vitro.

METHODSH9C2 cardiomyoblasts were cultured for 72 h in the presence of 0, 5.5, 12.5, 25 or 33 mmol/L glucose (with the osmotic pressure adjusted with monnitol). Semi-quantitative detection of GRK2 gene expression in H9C2 cardiomyoblasts was carried out using RT-PCR and phosph-Akt (Ser473) protein level was measured by Western blotting.

RESULTSGlucose in the culture medium (5.5 to 33 mmol/L) concentration-dependently increased the mRNA expression of GRK2 concentration and decreased phosphorylation Akt (ser473) level in in H9C2 cardiomyoblasts.

CONCLUSIONIncreased GRK2 gene expression may play an important role in cardiac dysfunction in diabetes.

Cell Line ; Diabetes Complications ; metabolism ; G-Protein-Coupled Receptor Kinase 2 ; genetics ; metabolism ; Gene Expression Regulation ; Glucose ; pharmacology ; Humans ; Myocytes, Cardiac ; cytology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Up-Regulation

OBJECTIVETo observe the characteristics of gene expression of type X collagen in the cartilage of end-plate and the fibrous annulus in the intervertebral disc of idiopathic scoliosis (IS) patients.

METHODInvestigating the expression of type X collagen in the peak disc and the lower end disc of 21 IS patients, the peak disc of 16 congenital scoliosis (CS) and the lumbar disc of 3 normal people (according with the principle of medical ethnics) by reverse transcript polymerase chain reaction.

RESULTSThe expression of type X collagen in the concave side of IS peak disc was higher than the convex side (P < 0.05). There was no significant difference of gene expression of type X collagen between the convex side and the concave side of the lower end disc (P > 0.05). The gene expression of type X collagen in the IS peak disc was higher than those of lower end disc (P < 0.05). For the CS peak discs, the expression of type X collagen of the concave side was higher than the convex side (P < 0.05).

CONCLUSIONThe expression of type X collagen of the IS peak disc increases, and the expression of type X collagen of the concave side is higher than the convex side. These changes may be secondary.

Adolescent ; Child ; Collagen Type X ; genetics ; metabolism ; Female ; Gene Expression ; Humans ; Intervertebral Disc ; metabolism ; Male ; Scoliosis ; genetics ; metabolism