Objective To investigate the expression of hypoxia-inducible 1 alpha(HIF-lα)and glucose transporter 1(Glut1)in human breast cancer and its relationship to proliferating cell nuclear antigen (PCNA)protein and clinical pathologic factors.Methods Immunohistochemical staining was used to measure the expression of HIF-lα.Glut1 and PCNA in human breast fibroadenoma,usual hyperplasia and breast carcinoma.Results HIF-1α expression was not found in breast fibroadenoma and hyperplastic Iesions.In contrast.the positive rate of HIF-1α was found in the ductal carcinoma in situ 55%(DCIS,11/20)and the invasive breast carcinoma 85%(51/60).Glut1 positivity in breast carcinoma was 58.8%(47/80).The totsl positive rate of PCNA in breast carcinoma was 75%(60/80),that in DCIS was 65%(13/20)and that in invasive carcinoma was 78.3%(47/60).There was a positive correlation between HIF-lα and Glut 1 level (r=0.653,P＜0.01),a positive correlation between HIF-1α and PCNA level(r=0.693,P＜0.01);and also a positive correlation between Glutl and PCNA level(t=0.742.P＜0.01).conclusion The overexpression of HIF-lα and its target gene Glut1 played important roles in carcinogenesis and progression of breast carcinoma and closely correlated with cell proliferation of breast carcinoma and may become a new target for treatment of breast carcinoma.

Objective To investigate the morphological changes in the sciatic nerve and the dorsal root ganglions (DRGs) and also gene expression in DRGs after non-freezing cold injury, and to explore the molecular mechanism of peripheral nerve cold injury and regeneration. Methods Twenty-four male Wistar rats were used. The sciatic nerve on one side was cooled to 4℃ for 2 h, and the sciatic nerve on the opposite side was exposed, but without cooling. Sciatic nerves and L4, L5 and L6 DRGs from both sides were harvested at the 1st, 2nd and 3rd week after cooling. Any pathological changes were observed using light and electron microscopy. Laser capture microdissection (LCM) was used to investigate the DRG neurons' gene expression. The array result was verified with RT-PCR for eight genes. Results Large fiber degeneration was obvious by the 7th day after cooling. Myelinated fiber regeneration had begun by the 14th day, so this time was chosen to explore the neurons' gene expression. Ninety-six genes and expressed sequence tags (ESTs) were up-regulated greater than 2 fold. Their proteins' functions were classified as adaptive response to external stimulus, apoptosis regulation, cell adhesion, immune and inflammation response,nerve regeneration, pain associated molecules, microtubule cytoskeleton, ion-channels, neurotransmitters and receptors, and neuropeptides. Conclusions A complex molecular mechanism is involved in cold injury and regeneration of the sciatic nerve, and many genes are involved. Large scale microarray analysis is a potent means to screen out related genes, thus suggesting future repair strategies.

Objective To investigate the neuroprotection effect of puerarin injection against H2O2 induced injury on hippocampal neurons.Methods The cells is divided into four groups,untreated cells as blank group,puerarin treated cells as negative control group,H2O2 treated cells as positive control group,puerarin(100 mg · L-1) treatment prior to H2D2 (40 μ mol · L-1) treated cells as experimental group.To find out optimal H2O2 working condition of positive control group,and the best puerarin concentration was determined by MTT assay.After 1 h H2O2 exposure,cell morphology was observed by microscope.Meanwhile superoxide dismutase (SOD) and hydrogen peroxidase (HP) activity,malondialdehyd (MDA)and glutathione (GSH) content and reactive oxygen species (ROS) level were measured by biochemical methods.The expression of Bcl-2 and Bax mRNA were measured by RT-PCr Results The optimal H2O2 workingcondition of positive control group is 40 μmol · L-1 H2O2 for 1 h,which survival cell rate in positive control group was (69.67 ±4.93)％.And optimal pretreatment concentration of puerarin is 100 mg · L-1.Compared to the positive control group,the morphology in experimental group was much more better.Compared to the blank group with (100％),the HP,SOD and GSH in positive control group were (56 ± 4.58) ％,(60.33 ± 6.11) ％,(51 ± 7) ％;while MDA,ROS in positive control group were (152.33 ± 6.03)％,(149.67 ± 10.79)％,it was increased and decreased with significantly (all P ＜ 0.01).Compared to the positive control group,the HP,SOD and GSH in experimental group were (88.67 ± 4.73) ％,(89.33 ± 4.51) ％,(90.33 ± 3.21) ％;while MDA,ROS in experimental group were (108.00 ± 4.58)％,(111.00 ± 8.32)％,it was increased and decreased with significantly (all P ＜ 0.01).Compared to the blank group,expression of Bcl-2 mRNA and Bax mRNA in positive control group were 0.37 ± 0.06,2.64 ± 0.46,it was increased and decreased with significantly (all P ＜ 0.01).Compared to the positive control group,expression of Bcl-2 mRNA and Bax mRNA in experimental group were 0.75 ± 0.08,1.44 ± 0.08,it was increased and decreased with significantly(all P ＜ 0.01).Conclusion Pretreatment of puerarinmay protect H2O2 induced injury of hippocampal neurons through modulation of Bax and Bcl-2 expression.

Objective To evaluate the CT findings of lymphofollicular thymic hyperplasia in adult myasthenia gravis (MG).Methods The CT findings of thymus area of 134 adult patients with lymphofolficular thymic hyperplasia in MG were reviewed,all of them with surgically and histologically proven diagnosis,and compared with the CT findings of 165 normal subjects.Results In the group of patient,CT showed enlargement of thymus in 31 patients,5 patients had nodule or mass(＜3 cm);thus 36 cases(26.9%)can confirmed diagnose by CT with thymic hyperplasia.CT showed 2 masses(＞3 em) and 9 patients(6.7%)had normal size thymus with soft-tissue density,it can considered with thymic hyperplasia.The spotty or streak shadow showed in other patients,though it could not be certain diagnosed as thymic hyperplasia,but could not be except it.The thymus area tissue complete replacement by fatty density were not found in patient group.The CT findings of patients had marked difference when compared with group of normal subjects(P＜0.01),except the spotty or streak shadows.Conclusion CT scan is an important method in diagnosing thymic lymphofollicular hyperplasia of MG in adult.

Objective To establish a method of measuring adipose content and fat distribution of the thigh in normal glucose tolerance (NGT) subjects,and to investigate its relation to insulin resistance.Methods Insulin sensitivity was evaluated by hyperinsulinemic and euglycemic clamp technique,and femoral adipose content and fat distribution were determined by MRI in 30 individuals with NGT including 15 with normal weight and 15 overweighted or obese subjects.Results Compared to normal weight group,the subscutaneous adipose tissue of thigh (SCAT) [(176.7?21.6) cm~2 vs (115.0?12.8 ) cm~2,P＜0.05],adipose tissue of thigh beneath the fascia (SFAT) [(75.4?4.4 ) cm~2 vs (57.5?4.7 ) cm~2,P＜0.01] and intermuscular adipose tissue (IMAT) [(28.3?3.2) cm~2 vs (14.5?1.1 ) cm~2,P＜0.01] were greater in overweight/obesity group.Overweight/ obesity group had lower insulin sensitivity( glucose disposal rate under steady state of hyperinsulinemic euglycemic clamp:4.54?0.43 vs 7.88?0.75,P＜0.01).SFAT and IMAT were significantly correlated with insulin sensitivity.SFAT showed the most marked correlation with insulin sensitivity.Multiple stepwise regression analysis showed that the increased SFAT played a pivotal role in insulin resistance.Conclusion The adipose content and fat distribution are highly correlated with insulin sensitivity and the adipose tissue of thigh beneath fascia may play the most significant role in insulin sensitivity.

OBJECTIVETo evaluate the early operative treatment and clinical results of pelvic fractures associated with urethra disruption.

METHODSFrom January 1995 to January 2005, 25 patients suffered from pelvic fractures combined urethra disruption treated by operation were retrospectively analyzed. According to Tile's classification, 1 case was stable pelvic fracture, 17 rotational unstable fractures, and 7 rotational combined vertical unstable fractures. The complete urethra rupture were in 23 cases and incomplete in 2 cases. The operative methods included: (1) emergency open reduction and internal fixation of the pelvis combined primary urethra suturing in 2 cases, partial suturing after realignment in 4 cases, realignment in 2 cases, and urethrovaginal penetrating wound repairing in 1 case; (2) primary urethra realignment only and delayed (range, 7 to 21 days) pelvic internal fixation in 10 cases; (3) early cystostomy and delayed (range, 3 to 21 days) urethra realignment and pelvic internal fixation in 6 cases.

RESULTSThe mean follow-up time of all patients was 34 months (range from 6 to 120 months). According to Majeed's evaluation, 17 cases of pelvic injury showed excellent results, 5 good, and 3 fare. After urinary catheter removed, the mean maximal urine flow rate of 19 (76%) patients was 18.6 ml/s and the mean scar length between both disrupted ends on the film of excretion urethrography was 0.51 cm. Five (20%) cases suffered in dysuria needed urethral dilatation or further surgery. One (4%) female could not control urination who need a second-look operation. The primary suprapubic soft tissue avulsion wound infection secondary to retropubic abscess was found in 1 case, posterior urethra-stenosis in 5 cases, sexual impotence in 3 cases, and incontinence in 1 case.

CONCLUSIONSThe satisfactory reduction and effective fixation of the pelvic fractures is an anatomical basis for receiving "tension-free urethral anastomosis".

Adult ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; injuries ; Postoperative Complications ; prevention & control ; Retrospective Studies ; Time Factors ; Treatment Outcome ; Urethra ; injuries

OBJECTIVETo observe the protective effects of Tongxinluo (TXL) on apoptosis of rat cardiac microvascular endothelial cells (RCMECs) resulting from homocysteine (Hcy) induced endoplasmic reticulum stress (ERS), and to determine the signaling pathway behind its protection.

METHODSPrimary cultured RCMECs were isolated from neonatal rats using tissue explant method. The morphology of RCMECs was observed using inverted microscope, identified and differentiated by CD31 immunofluorescence method. Selected were well growing 2nd-4th generations of RCMECs. The optimal action time was determined by detecting the expression of glucose regulated protein 78 (GRP78) using immunofluorescence method. In the next experiment RCMECs were divided into 5 groups, i.e., the blank control group, the Hcy induced group (Hcy 10 mmol/L, 10 h), the Hcy + TXL group (Hcy 10 mmol/L + TXL 400 µg/mL), the Hcy +LY294002 group (Hcy 10 mmol/L + LY294002 5 µmol/L, LY294002 as the inhibitor of PI3K), the Hcy + LY294002 + TXL group (Hcy 10 mmol/L + LY294002 5 µmol/L + TXL 400 µg/mL). The apoptosis rate of RCMECs was detected by flow cytometry. mRNA and protein expressions of GRP78, C/ EBP homologous protein (CHOP), and cysteinyl aspartate specific proteinase-12 (caspase12) were detected by real-time reverse transcription PCR (RT-PCR) and Western blot respectively. Expression levels of phosphorylation of phosphatidylinositol 3-kinase (P-PI3K), total phosphatidylinositol 3-kinase (T- P13K) , phosphorylation of kinase B (P-Akt) , and total kinase B (T-Akt) were detected by Western blot.

RESULTSTen hours Hcy action time was determined. Compared with the blank control group, the apoptosis rate was increased (22.77%), mRNA and protein expressions of GRP78, CHOP, and Caspase-12 were increased, protein expressions of P-PI3K and P-Akt,ratios of P-PI3K/T-PI3K and P-Akt/T-Akt were decreased in the Hcy induced group (P < 0.05, P < 0.01). Compared with the Hcy induced group, the apoptosis rate was decreased (10.17%), mRNA and protein expressions of GRP78, CHOP, and Caspase-12 were decreased, and expression levels of P-PI3K, P-Akt, P-PI3K/T-PI3K, and P-Akt/T-Akt were increased in the Hcy + TXL group (P < 0.05, P < 0.01). Compared with the Hcy + TXL group, the apoptosis rate was increased (17.9%), mRNA and protein expressions of GRP78, CHOP, and Caspase-12 were increased, expression levels of P-PI3K and P-Akt, ratios of P-PI3K/T-PI3K and P-Akt/T-Akt were decreased in the Hcy + TXL + LY294002 group (P < 0.05, P < 0.01).

CONCLUSIONTXL could inhibit the apoptosis of RCMECs resulting from Hcy-induced ERS and its mechanism might be associated with activating PI3K/Akt signaling pathway.

Animals ; Apoptosis ; drug effects ; Caspase 12 ; metabolism ; Cells, Cultured ; Chromones ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Endoplasmic Reticulum Stress ; Endothelial Cells ; drug effects ; Morpholines ; pharmacology ; Myocardium ; cytology ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Signal Transduction ; Transcription Factor CHOP ; metabolism

Objective To evaluate the effect of clopidogrel premedication on in-hospital major adverse cardiovascular and cerebral events (MACCE) and bleeding outcomes before coronary artery bypass graft surgery (CABG). Method A total of 2021 patients who underwent CABG from July 2003 to September 2005 were divided into either clopidogrel ( n = 479) or no clopidogrel (1542) group before CABG. Patients with clopidogrel administration ( n = 479) were subdivided into < 5 d ( n = 154) ,5～7d(n = 183)and >7d(n = 142) group according to timing of clopidogrel withdrawal before surgery. In-hospital MACCE and perioperative bleeding outcomes were analyzed among groups. Results Patients who took clopidogrel before surgery had nonsignificantly rates of bleeding and in-hospital MACCE compared with those patients not administered clopidogrel. No differences were found about the incidence of total bleeding,minor bleeding,transfusions of red blood cells,fresh frozen plasma,whole blood and in-hosptial MACCE among three subgroups.The < 5 d group had higher incidence of major bleeding and more platelets transfusions than 5 ～ 7 d [47.8% vs. 31.9%,P < 0.017; (0.08 ±0.38) U vs. (0.00±0.00) U,P <0.017,respectively]and >7 d group [47.8% vs. 20.3%,P <0.017; (0.08±0.38) U vs. (0.00±0.00) U,P <0.017,respectively). However,there were no significant differences between 5 ～ 7 d and > 7 d group ( P > 0.05). Conclusions Gopidogrel administration before CABG does not increase the incidence of in-hospital MACCE events. However,the perioperative risk of bleeding will rise if the patients withhold clopidogrel less than five days before surgery.

Fifteen known compounds were isolated from Swertia delavayi by silica gel, Sephadex LH-20 and Rp-18 column chromatographies. Based on extensive spectroscopic analysis (MS, 1H, 13C-NMR), their structures were identified aserythrocentaurin (1), erythrocentaurindimethylacetal (2), sweroside (3), swertiamarin (4), gentiopicroside (5), swertiakoside A (6), 2'-O-acetylswertiamarin (7), 4'-O-[(Z) -coumaroyl] swertiamarin (8), 1,5,8-trihydroxy-3-methoxyxanthone (9), 8-O-β-D-glucopyranosyl-1-hydroxy-2,3, 5-trimethoxyxanthone (10), 8-O-[β-D-xyl- opyranosyl-(1 --> 6)-β-D-glucopyranosyl]-7,8-dihydroxy-3-methoxyxanthone (11), isovitexin (12), β-sitosterol (13), daucosterol (14), and oleanolic acid (15). Among them, ten ones (14, 7-11, 13) were obtained from S. delavayi for the first time. The isolates were evaluated for their anti-HBV activities in HepG 2. 2. 15 cell line in vitro. The results showed that compound 1, 2, 6, 7, 9 and 12 exhibited significant inhibitory activity on HBV DNA replication with IC50 values from 0.05 to 1.46 mmol x L(-1).
Antiviral Agents ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Hepatitis B virus ; drug effects ; genetics ; Magnetic Resonance Imaging ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Swertia ; chemistry

OBJECTIVETo identify the RUNX2 gene mutation in two unrelated Chinese families with cleidocranial dysplasia (CCD), and to assess the feasibility of gene diagnosis for patients with CCD.

METHODSGenomic DNA was isolated from peripheral blood samples of 4 patients and 4 healthy members in the two pedigrees as well as 102 unrelated healthy controls. All 7 coding exons and their flanking intronic sequences of the RUNX2 gene were amplified by PCR, then the PCR products were sequenced bi-directionally. The sequencing results were compared with normal sequences in GenBank to identify the mutation. The mutation was confirmed by RFLP with restriction endonuclease.

RESULTSIn one family, a novel heterozygous missense mutation c.346T to A (W116R) in exon 1 of the RUNX2 gene was detected in the two affected individuals, and the mutation was further confirmed with Bsr I restriction endonuclease digestion. In the other family, a novel nonsense mutation c.610A TO T (K204X) was identified in the two patients. No above sequence change was found in the 102 healthy controls.

CONCLUSIONTwo novel RUNX2 mutations were found in two unrelated Chinese families with cleidocranial dysplasia. The identification of these mutations further extended the mutation spectrum of RUNX2 gene and will facilitate prenatal diagnosis and gene diagnosis of CCD.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Cleidocranial Dysplasia ; genetics ; physiopathology ; Core Binding Factor Alpha 1 Subunit ; genetics ; DNA Mutational Analysis ; Female ; Humans ; Infant ; Male ; Mutation ; Pedigree ; Restriction Mapping