Objective To explore the effect of wild type or mutant parkin gene expression on the growth of human hepatocellular carcinoma cell line Huh-7. Methods The parkin (wild type or mutant) expression vector and empty vector were transferred into Huh-7 cell lines with LipofectAMINE 2000 reagents. The positive clones that expressed parkin gene stably were chosen by G418 and checked by reverse transcription-polymerase chain reaction (RT-PCR) to check the DNA sequences. The cytobiological behaviors of those positive clones were analyzed by cell proliferation assay and tumorigenesis in nude mice. Results Huh-7 cell lines that expressed wild type or mutant parkin gene stably were successfully established. The growth of wild type parkin-expressed cells was obviously inhibited compared with the control cells transfected with empty vectors(t= 3. 875, P= 0. 031).The volume of tumor formed by wild type parkin-expressing cells in nude mice was also significantly reduced (t=8. 228,P=-0. 003). Mutant parkin gene expression had a slight effect on the growth of Huh-7 cells in vitro and in vivo (P＞0.05). Conclusion The re-expression of wild type parkin gene can favor the malignant phenotype revision of Huh-7 cells. Therefore, it might be a good candidate for tumor suppressor gene associated with HCC.

Glomerular hypertrophy is the main pathological characteristic in the early stage of diabetic nephropathy (DN), and its regulatory mechanism is closely related to mammalian target of rapamycin (mTOR) signaling pathway activity. mTOR includes mTOR complex 1 (mTORC1) and mTOR complex 2(mTORC2), in which, the upstream pathway of mTORC1 is phosphatidylinositol-3-kinase (PI3K)/serine-threonine kinase(Akt)/adenosine monophosphate activated protein kinase(AMPK), and the representative signaling molecules in the downstream pathway of mTORC1 are 4E-binding proteins(4EBP) and phosphoprotein 70 S6Kinase(p70S6K). Some Chinese herbal extracts could improve cell proliferation via intervening the expressions of the key molecules in the upstream or downstream of PIK/Akt/mTOR signaling pathway in vivo. As for glomerular mesangial cells(MC) and podocyte, mTOR plays an important role in regulating glomerular inherent cells, including adjusting cell cycle, energy metabolism and matrix protein synthesis. Rapamycin, the inhibitor of mTOR, could suppress glomerular inherent cell hypertrophy, cell proliferation, glomerular basement membrane (GBM) thickening and mesangial matrix deposition in model rats with DN. Some Chinese herbal extracts could alleviate glomerular lesions by intervening mTOR signaling pathway activity in renal tissue of DN animal models or in renal inherent cells in vivo and in vitro.
Animals ; Diabetic Nephropathies ; drug therapy ; enzymology ; genetics ; pathology ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypertrophy ; drug therapy ; enzymology ; genetics ; pathology ; Kidney Glomerulus ; drug effects ; metabolism ; pathology ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; genetics ; metabolism

Objective To observe the effect of splenectomy on mortality and brain water content of rats with brain injury so as to explore novel way for better clinical management of patients with severe traumatic brain injury. Methods Adult male SD rats were randomly divided into three groups, ie, sham operation on brain and spleen (Group A, n = 23), experimental brain trauma & sham operation on spleen (Group B, n =48) and experimental brain injury & splenectomy (Group C, n = 47). Modified Feeney' s method was used to create the animal model of experimental brain trauma, Longa' s scale was applied to evaluate the neurologic defect. Mortality within seven days following brain injury was calculat-ed. In the meantime, the brain water content was detected at days 1 (n = 8), 2 (n = 8), 3 (n = 8) and 7 (n = 7) after brain injury in each group, Results No statistical difference of Longs' s scale was found between Group B and Group C (P > 0.05). The mortalities within seven days after brain injury were 0%, 35.42 and 14.89% in Groups A, B and C respectively, with statistical difference between groups (P<0.05). The brain water content of Groups B and C at days 1, 2, 3 and 7 were (81.98±0.35)% & (81.78±0.41)%, (82.58±0.63)% & (81.81±0.48)% (P<0.05),(82.54±0.54)% & (81.52±0.84)% (P<0.05) and (81.50±0.41)% & (81.21±0.36)% (P>0.05) respectively. Conclusion Splenectomy can effectively reduce brain water content and significantly decrease mortality in rata with brain injury.

BACKGROUNDThe abnormalities of coronary arteries, though rare and sometimes benign, may first present clinically as myocardial infarction or sudden death. Multi-detector computed tomography (MDCT) is a non-invasive test that is highly suitable for detecting these anomalies. The study aimed to review the 64-MDCT appearance of the coronary artery anomalies in 66 patients and to discuss the clinical importance of these anomalies.

METHODSIn 6014 consecutive patients examined over 12 months by 64-MDCT for the study of coronary artery disease, 66 were diagnosed for coronary artery anomalies. All patients were symptomatic for one or more of the following diseases: chest pain, dyspnoea, palpitations, arrhythmia and myocardial infarction. Nine patients had undergone a coronary angiography. All the CT images were evaluated by two radiologists and one cardiologist. The right coronary artery (RCA) and the conus branch arising separately, myocardial bridging and duplication of arteries were not analysed in our study.

RESULTSThe incidence of coronary artery anomalies found in our study group was 1.097%. In the selected patients, seven different types of coronary anomalies were found by 64-MDCT examination. The high takeoff, origin of the coronary artery from the opposite or noncoronary sinus with an anomalous course, and coronary artery fistula were the three common forms of anomalies (n = 16, 18 and 16, respectively). Compared with the results of the coronary angiography, the number of the drainage sites of two coronary artery fistula was less in MDCT images (3 small sites in total). In all cases, coronary artery computed tomography angiography (CTA) technique was able to recognize the origin of the coronary artery, its three-dimensional course and its spatial relationship with the adjacent structures. Conventional coronary angiography in two cases, however, was unable to provide sufficient information for correct and complete diagnosis.

CONCLUSIONSIn conclusion, the study showed that 64-MDCT, especially the volume rendering technique (VRT), may be useful for the assessment of complex variations, even if the conventional angiography may not be sufficient. It may be considered as the first-choice imaging modality when an anomalous coronary artery is suspected.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Coronary Vessel Anomalies ; diagnostic imaging ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Young Adult

OBJECTIVESTo study the etiology of persistent hematospermia and to evaluate the efficacy of transrectal ultrasonography (TRUS)-guided transperineal needle aspiration and irrigation for diagnosis and treatment of persistent hematospermia.

METHODSTwelve patients were included in the study, with a mean age of (36.4 +/- 10.8) years old, and a mean duration of the disease of (13.9 +/- 6.4) months. After the expressed prostatic secretion (EPS) by prostatic massage was cultured, patients with recurrent hematospermia received TRUS-guided transperineal needle aspiration for seminal vesicle fluid (SVF), which was sent for bacteriological and cytological examination. If the EPS culture were positive, certain antibiotics according to the drug sensitivity assay were injected into the abnormal seminal vesicle(s) via TRUS-guided transperineal needle puncture. The treatment would be repeated one month later if the patients still had hematospermia. The patients were followed up every three months.

RESULTSAbnormal images were found in left seminal vesicle (SV) in 4 cases, right in 3 cases, bilateral in 2 cases, and no abnormal findings in 3 cases. The abnormal findings included: 7 cases of SV and/or ejaculatory duct dilation, 3 cases of thickening SV wall, 3 cases of calcification or calculi of SV, and 1 case of Müllerian duct cyst. SVF cultures were positive in 7 cases: methicillin-resistant Staphylococcus aureus (MRSA) 4 cases, methicillin-resistant coagulase-negative Staphylococcus (MRCNS), E. Coli, Proteus mirabilis 1 case, respectively. In five of these 7 cases, bacteriological cultures of SVF and EPS showed the same results. All patients were treated by TRUS-guided transperineal injection of certain antibiotics into SV. Seven cases were injected once, 5 cases twice. The mean follow-up period of 10 patients was (16.7 +/- 5.9) months. Hematospermia disappeared in 6 cases.

CONCLUSIONSSV infection of bacteria, especially infection of the drug resistant strains was one of the main causes of persistent hematospermia. The difficulties in treatment of persistent hematospermia were due to infection of drug resistant bacteria, calcification or calculi of SV, obstruction of ejaculatory duct. TRUS-guided transperineal aspiration of SVF was helpful to the etiologic diagnosis of persistent hematospermia.

Adult ; Drug Resistance, Bacterial ; Genital Diseases, Male ; etiology ; microbiology ; Humans ; Male ; Middle Aged ; Pilot Projects ; Seminal Vesicles ; microbiology ; Ultrasound, High-Intensity Focused, Transrectal

OBJECTIVETo study the expression and distribution of bone morphogenetic protein (BMP) in newly formed bone by distraction osteogenesis (DO), and to explore the mechanism of the DO bone formation and remodeling.

METHODSThe cleft palate (CP) experimental animal models (23 Rhesus monkeys) were established surgically. In experimental group (21 Rhesus monkeys), the palatal defects were corrected by means of DO at the rhythm of 0.4 mm twice per day. The specimens were retrieved under euthanasia at 1, 2, 4, 6, 8, 12, 24 weeks intervals respectively in retention period. BMP immunohistochemical study was then performed. The blank control and experimental group (each of 2 animals) were set for comparison study.

RESULTSThe immunohistochemical study showed that BMP existed mainly in cytoplasma of osteoblasts, during the process of new bone formation. In early stage of 1 or 2 weeks, abundant osteoblasts aggregating on surfaces of the new bone trabeculae with positive DAB dye were observed. Through 4 to 6 weeks, the proliferative osteoblasts with very strong positive DAB dye indicating BMP expression were recorded. From 8 to 12 weeks, the expression of BMP and quantity of osteoblasts decreased gradually while more matured new bone structures were observed.

CONCLUSIONDuring the whole retention period, the expression of BMP showed a tendency from weak to strong and then to final cessation, this indicated a process of formation, remodeling and maturation of osteogenesis.

Animals ; Bone Morphogenetic Proteins ; metabolism ; Cleft Palate ; metabolism ; surgery ; Macaca mulatta ; Osteogenesis, Distraction

OBJECTIVETo investigate the correlation of the degrees of intravesical prostatic protrusion (IPP) measured by transabdominal ultrasound with the clinical evaluation parameters in BPH patients.

METHODSWe measured the length of IPP in the mid-sagittal section by transabdominal ultrasound in 275 BPH outpatients with lower urinary tract symptoms, and analyzed the correlations of the degree of IPP with the age, prostatic volume (PV), international prostatic symptom score (IPSS), maximum uroflow rate (Qmax), and postvoid residual urine volume (PVR) of the patients.

RESULTSThe degree of IPP was correlated positively with the age (r = 0.210, P < 0.01), PV (r = 0.534, P < 0.01) and PVR (r = 0.314, P < 0.01), but negatively with the Qmax (r = -0.364, P < 0.01) of the BPH patients. There was no significant correlation between the degree of IPP and IPSS (r = 0.064, P = 0.299).

CONCLUSIONThe degree of IPP may be associated with the age and prostatic volume of BPH patients. Transabdominal ultrasound measurement of IPP is a useful noninvasive method to assess the presence and severity of bladder outlet obstruction in BPH patients.

Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Prostate ; diagnostic imaging ; Prostatic Hyperplasia ; diagnostic imaging ; Ultrasonography ; Urinary Bladder Neck Obstruction ; diagnostic imaging

BACKGROUNDDNA immunization is a promising novel type of immunotherapy against allergy. An estimated 79.2% patients with asthma, wheezing and/or rhinitis suffer from Dermatophagoides pteronyssinus group 2 (Der p 2) allegen. The aim of the present study was to determine whether DNA vaccine encoding Der p 2 could generate immunologic protection in recombinant Der p 2 (rDer p 2) allergen-induced allergic airway inflammation mice model and to understand the role of DNA vaccination in specific-allergen immunotherapy for asthma.

METHODSAfter DNA vaccination, BALB/c mice were sensitized by intraperitoneal injection (i.p) and challenged by intranasal instillation of rDer p 2. The lung tissues were assessed using hematoxylin and eosin. Mucus-producing goblet cells were identifed using periodic acid-Schiff (PAS)/alcian blue. The total cell number and composition of bronchoalveolar lavage samples were determined. The levels of the cytokines IL-4 and IFN-gamma, as well as IgE and IgG2a in the serum were determined by enzyme-linked immunosorbent assay. Allergen-specific IL-4 and IFN-gamma production by spleen cells were also measured by enzyme-linked immunosorbent assay. Expression of signal transducer and activator of transcription 6 (STAT6) in splenocytes were determined by Western blot.

RESULTSDNA vaccine encoding Der p 2 allergen inhibited extensive infiltration of inflammatory cells and production of mucin induced by allergen. The influx of eosinophils into the lung interstitium was significantly reduced after administration of DNA vaccine. Significant reductions of IL-4 and increase in levels of IFN-gamma in bronchoalveolar lavage fluid were observed. The allergen-specific IgE was markedly decreased in mice receiving DNA vaccination. Allergen could induce higher IFN-gamma, weaker IL-4 in cultured spleen cells from mice receiving DNA vaccine. DNA vaccination inhibited STAT6 expression of spleen cells induced by allergen.

CONCLUSIONThese results indicated that DNA vaccine encoding Der p 2 allergen generates immunologic protection in recombinant Der p 2 allergen-induced allergic airway inflammation mice model with regulating the immune response towards a Th1-type reaction.

Animals ; Antigens, Dermatophagoides ; genetics ; immunology ; Arthropod Proteins ; Asthma ; immunology ; therapy ; Eosinophilia ; prevention & control ; Humans ; Immunoglobulin E ; blood ; Immunoglobulin G ; blood ; Interferon-gamma ; biosynthesis ; Interleukin-4 ; biosynthesis ; Mice ; Mice, Inbred BALB C ; STAT6 Transcription Factor ; Th1 Cells ; immunology ; Trans-Activators ; analysis ; Vaccination ; Vaccines, DNA ; immunology

OBJECTIVETo study the change of tissue oxygen index (TOI) by non-invasive near infrared spectroscopy (NIRS) and to investigate the blood flow variety of fibula flaps after operation.

METHODSThirty-six patients who accepted fibula flap reconstruction were chosen as subjects. Authors measured the TOI of the fibula flaps and the control side every four hours in the first twenty-four hours, and measured these positions with the intermittence of twelve hours from second to eighth day after operation.

RESULTSThirty-five flaps were successful and one failed. The TOI of fibula flaps shortly after operation was significantly lower than that before the operation( P<0.05). In the successful cases the TOI of fibula flaps and the control sides was fluctuated from 50.0% to 72.0%. The TOI of fibula flaps was significantly lower than that of the control sides within 144 hours after operation(P<0.05). After 144 hours the TOI was equal to that of the control side. TOI of the failed case decreased dramatically.

CONCLUSIONSNIRS can reliably indicate the change of TOI in buried flaps and detect ischemia at the early stage. The TOI of the fibula flaps depress at the early stage and returns to normal at 144 hours after operation.

Adolescent ; Adult ; Aged ; Female ; Fibula ; transplantation ; Humans ; Male ; Middle Aged ; Oximetry ; methods ; Oxygen ; blood ; Spectroscopy, Near-Infrared ; methods ; Surgical Flaps ; blood supply ; Young Adult

OBJECTIVETo investigate the expression and regulation mechanism of mismatch repair (MMR) genes in chronic myeloid leukemia (CML).

METHODSExpression of MMR genes hMSH2, hMSH3, hMSH6, hMLH1 and hPMS2 mRNAs in 62 CML patients and K562 cell line were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Expression of bcr-abl mRNA and MMR genes mRNA were detected by RT-PCR in 26 CML patients with allogeneic peripheral blood stem cell transplantation (allo-PBSCT) and 4 CML patients on imatinib treatment. Expression of bcr-abl mRNA was detected by RT-PCR and tyrosine phosphorylation of BCR-ABL fusion protein by Western blot.

RESULTSExpression of hMSH2, hMSH3 and hMLH1 mRNA was significantly lower in CML and K562 cells than in normal control (P < 0.05). In 26 CML with allo-PBSCT and 4 CML patients on imatinib treatment, expressions of hMSH2, hMSH3 and hMLH1 mRNA was enhanced while expression of bcr-abl mRNA decreased. In CML MNC after imatinib treatment and in K562 cells, expression of hMSH2, hMSH3 and hMLH1 mRNA was enhanced while tyrosine phosphorylation of BCR-ABL fusion protein decreased.

CONCLUSIONExpressions of hMSH2, hMSH3 and hMLH1 mRNA were down-regulated by bcr-abl fusion gene.

Adult ; Aged ; Antineoplastic Agents ; pharmacology ; Benzamides ; DNA Mismatch Repair ; Female ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Humans ; Imatinib Mesylate ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Middle Aged ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction