OBJECTIVETo observe the effect of Yijing Recipe (YR) on the apoptosis of testis spermatogenic cells and the protein expression of Bcl-2/Bax in rats with adenine induced infertility.

METHODSTotally 75 Wistar rats were randomly divided into 5 groups, i.e., the blank control group, the model group, the high dose YR group, the middle dose YR group, and the low dose YR group, 15 in each group. Except those in the blank control group, rats in the rest groups were intragastrically administered with adenine for 10 successive days. From the 11th day, rats in the blank control group and the model group were fed with equal volume of normal saline. Rats in the YR groups were intragastrically administered with YR at different doses (3.38 g/100 g; 1.69 g/100 g; 0.85 g/100 g), once daily for 20 consecutive days. All rats were killed by the end of the experiment and their testes extracted. The apoptosis of spermatogenic cells and the expression of Bcl-2/Bax proteins were detected by terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) and SABC method.

RESULTSCompared with the blank control group, the Bcl-2 protein expression decreased, the Bax protein expression increased, and the apoptosis index increased in the model group, showing statistical difference (P <0.01). Compared with the model group, the Bcl-2 protein expression increased in the three YR treated groups (P <0.01, P <0.05). The Bax protein expression level decreased in the high and middle dose YR groups (P <0. 01, P <0. 05). The apoptosis index decreased in the middle dose YR group (P <0.01).

CONCLUSIONYR could inhibit the apoptosis of spermatogenic cells through regulating the expression of Bcl-2 and Bax protein in the testis.

Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Infertility ; metabolism ; Male ; Rats ; Spermatogenesis ; drug effects ; Testis ; metabolism ; bcl-2-Associated X Protein ; metabolism

China ; Crystallography, X-Ray ; Light-Harvesting Protein Complexes ; chemistry ; metabolism ; Photosynthesis ; Spinacia oleracea ; chemistry ; metabolism

Activation of microglia plays a vital role in the initiation and maintenance of specific neuropathic pain states. By activating microglia in central nervous system, Toll-like receptor 4 (TLR4) can promote the release of proinflammatory cytokines and neuroactive compounds, participate in the initiation and maintenance of neuropathic pain, and trigger the opiate side effects. Therefore, TLR4 may be a potential therapeutic target for neuropathic pain. Inhibition of TLR4 has shown some biological effects in neuropathic pain models and ibudilast (the TLR4 pathway-inhibiting agent) has been approved for for phase 2 clinical trials. This article briefly reviews the structure, function, and mechanism of TLR4 as well as the development of TLR4-targeted drugs.
Humans ; Neuralgia ; drug therapy ; physiopathology ; Toll-Like Receptor 4 ; antagonists & inhibitors ; physiology

Objective To investigate the curative effect of YU Yun pulse-feeling-based acupuncture therapy for the treatment of middle-late liver cancer. Methods A total of 60 middle-late liver cancer patients were divided into control group and treatment group by stratified randomization method,30 cases in each group. The control group was given integrated Chinese and western medicine therapy according to the clinical pathway, including anti-cancer treatment such as vascular interrention,molecular targeted therapy,chemotherapy,radio therapy, and focal ablation therapy,as well as chinese medicine treatment based on disease differentiation and syndrome differentiation. And the treatment group was given YU Yun pulse-feeling-based acupuncture therapy on the basis of treatment for the control group. The two groups received 12-week treatment, and then their curative effects were compared. Results The treatment group had better effect on increasing the survival rate, prolonging survival time, improving the scores of clinical symptoms, stabilizing tumor size, and increasing the scores of Karnofsky Performance Status (KPS)than the control group,the difference being significant (P < 0.05 or P <0.01). Conclusion YU Yun pulse-feeling-based acupuncture therapy exerts certain curative effect for the treatment of middle-late liver cancer.

OBJECTIVETo prepare pneumolysin as a new protein carrier of vaccine against otitis media with genetic engineering technology and establish the base of the study on pneumococcal conjugative vaccines.

METHODSGenomic DNA was isolated from streptococcus pneumoniae. A pair of primers which included two restriction sites was designed based on the published pneumolysin gene sequence. The pneumolysin gene was amplified from pneumococcal DNA with PCR technology. The restriction enzyme digested fragment was linked into the cloning vector PET-28a and the recombinant plasmid DNA containing pneumolysin was then transfected into host cell E. coli JM109 (DE3).

RESULTSDNA fragments were subcloned to construct the complete pneumolysin gene by a conventional coning and PCR. The inserted pneumolysin gene sequence was confirmed by DNA sequencing and the pneumolysin protein was successfully expressed. The relative molecular mass of the expressed product was 52 000. The expressed product amounted to 8% of the total host cell protein.

CONCLUSIONSThe pneumolysin gene was successfully cloned into host cell using genetic engineering technology. The recombinant pneumolysin was expressed and purified for preparation. This work laid a foundation of the preparation of pneumococcal conjugative vaccines.

Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Genetic Engineering ; Genetic Vectors ; Plasmids ; Pneumococcal Vaccines ; genetics ; Streptococcus pneumoniae ; genetics ; Streptolysins ; biosynthesis ; genetics

OBJECTIVETo explore the role of HOXB2 gene in the proliferation of primary human umbilical vein endothelial cells (HUVECs) and the protective effects of VEGF on the endothelia against radiation injury.

METHODSHUVECs were isolated, cultured, subcultured and identified. (1) Liposome coated oligodeoxynucleotide (odn) and homeoboxB2 antisense oligodeoxyncleotide (HOXB2asodn) were prepared prepared in the concentrations of 0.25, 0.5, 1.0 and 2.5 mg/L for the stimulation of HUVEC. (3)H-TdR incorporation test and MTT method were employed to determine the proliferation activity of HUVECs after activation. The cell cycle analysis of HUVECs was determined by flow cytometry. The expression level of HOXB2mRNA within HUVECs was detected by RT-PCR (reverse transcription polymerase chain reaction). (2) HUVECs were separately treated with the addition of VEGF in concentration of 50 microg/L, by radiation in the dose of 6 Gy or 12 Gy (60)Co gamma gamma ray, or radiation with 12 Gy (60)Co gamma gamma ray followed by the addition of VEGF in dose of 50 microg/L. The cellular morphology was observed and the cellular proliferation activity was determined by MTT method.

RESULTS(1) The proliferation activity of HUVECs could be markedly inhibited by liposome coated HOXB2asodn in comparison to liposome-odn (P < 0.05 or 0.001), and the inhibition effect was positively correlated with the increase in asodn concentration. The cell ratio in S phase and the expression level of the HOXB2mRNA could be lowered by asodn in dose of 2.5 mg/L (P < 0.05 or 0.001). (2) Radiation by (60)Co gamma ray could lead to the nuclear enlargement, vacuolation in the cytoplasm, multiplicity of nucleus and nuclear swelling. The proliferative activity of HUVECs was increased from 0.365 +/- 0.047 and 0.487 +/- 0.022 without radiation to 0.557 +/- 0.042 and 0.648 +/- 0.021 24 and 48 hours after 6 Gy radiation However it was decreased to 0.263 +/- 0.038 and 0.306 +/- 0.024 (P < 0.01) after 12 Gy (60)Co gamma ray radiation. Nevertheless, the cell morphology was obviously improved and the proliferation was enhanced by the addition of VEGF after 12 Gy radiation.

CONCLUSIONHOXB2 gene played important roles in the biological activities of HUVECs. Small dose (6 Gy) gamma-radiation could promote, but large dose (12 Gy) could decrease the mRNA expression of HOXB2 gene in HUVECs. In addition, VEGF could protect HUVECs against radiation injury.

Cell Proliferation ; drug effects ; radiation effects ; Cells, Cultured ; Endothelial Cells ; drug effects ; radiation effects ; Endothelium, Vascular ; cytology ; drug effects ; radiation effects ; Genes, Homeobox ; genetics ; Homeodomain Proteins ; genetics ; Humans ; Liposomes ; pharmacology ; Oligonucleotides, Antisense ; genetics ; Radiation Injuries ; Transcription Factors ; genetics ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factors ; pharmacology

OBJECTIVETo discuss the mechanisms of electroacupuncture at "Neiguan" (PC 6) on p38 MAPK signaling pathway in rats with cardiac hypertrophy.

METHODSForty SD rats were randomly divided into four groups: a normal group, a model group, a model plus p38 MAPK inhibitor group, a model plus electroacupuncture group, ten rats in each group. The model rats were established by subcutaneous injection 3 mg/(kg x d) of Isoprenaline Hydrochloride; model plus p38 MAPK inhibitor group were injected 0.3 mg/(kg x d) of specific inhibitor SB 203580; model plus electroacupuncture group was treated by electroacupuncture at "Neiguan"(PC 6) with continuous-wave, 2 Hz and 1 mA for 20 minutes, once a day for 14 days. There was no treatment in other two groups. The contents of TNF-alpha and IL-1beta in heart tissue were detected by radioimmunoassay and the p38 MAPK, p-p38 MAPK by western blot.

RESULTSCompared with normal group, the contents of TNF-alpha, IL-1beta, p38 MAPK, p-p38 MAPK were significantly increased in model group (all P < 0.01). The contents of TNFalpha, IL-1beta, p38 MAPK, p-p38 MAPK were significantly decreased in model plus p38 MAPK inhibitor group and model plus electroacupuncture group, compared with model group, all P < 0.05; compared with normal group, P < 0.05, P < 0.01; but no significant difference between model plus p38 MAPK inhibitor group and model plus electroacupuncture group (P > 0.05).

CONCLUSIONElectroacupuncture at "Neiguan" (PC 6) can prevent the phosphorylation of p38 MAPK of myocardial hypertrophy, and the mechanism maybe adjust p38 MAPK signaling pathway by inhibiting the expression of TNF-alpha and IL-1beta.

Acupuncture Points ; Animals ; Cardiomegaly ; metabolism ; therapy ; Electroacupuncture ; Interleukin-1beta ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Intracranial atherosclerotic disease (ICAD) of a major intracranial artery, including middle cerebral artery (MCA),basilar artery, is the most common causes of stroke and is associated with a high risk of recurrent stroke in China. The difficulty to treatment these high-risk disease is to identify high-risk stroke subgroups and to develop more effective treatments (aggressive medical therapy/endovascular therapy). With the benefits, including non-invasive, in vivo, and no-ionizing radiation, 3.0 Tesla high-resolution magnetic resonance imaging (HR MRI) could be used to stratify high-risk patients, monitor progression of disease, and evaluate clinical efficacy, based on MCA wall structure and plaque characteristic. HR MRI has the latency of predicting high-risk patients benefit from endovascular therapy, having a broad application prospect during psot-SAMMPRIS era. The current research on MCA stenosis using HR MRI focuses on methodoiogy, diagnosis and differential diagnosis, etiology, and lacks of clinical efficiency evaluation and prognostic analysis of ICAD treatment, especially lacks the research on in-stent restenosis, which needs further investigation.
Brain ; China ; Constriction, Pathologic ; Diagnosis, Differential ; Humans ; Intracranial Arteriosclerosis ; Magnetic Resonance Imaging ; Middle Cerebral Artery ; pathology ; Plaque, Atherosclerotic ; Prognosis ; Stroke ; Treatment Outcome

OBJECTIVETo evaluate the value of cardiac troponin I (CTnI) measurement in predicting anthracycline-induced cardiotoxicity in patients with breast cancer.

METHODSThis study was conducted among 186 breast cancer patients receiving anthracycline-based chemotherapy. Serum cTnI concentrations before and after each cycle of the chemotherapy and the left ventricular ejection fraction (LVEF) before and at the 2nd, 4th and 6th months of the treatment were recorded. According to serum cTnI concentration, the patients were divided into CTnI+ group (with serum CTnI concentration of no less than 0.1 ng/ml, n=60) and CTnI- (<0.1 ng/ml) group (n=126).

RESULTSNo patients in this series experienced cardiac heart failure (CHF). The number of patients with a LVEF reduction by over 10% from the baseline was 16 (26.7%) in CTnI+ group, as compared to 7 (5.6%) in CTnI- group, showing a significant difference between the two groups (P<0.01).

CONCLUSIONCTnI can be a useful marker for early prediction of anthracycline-induced cardiotoxicity in patients with breast cancer.

Adult ; Aged ; Anthracyclines ; adverse effects ; therapeutic use ; Antibiotics, Antineoplastic ; therapeutic use ; Biomarkers ; blood ; Breast Neoplasms ; drug therapy ; Cardiotoxins ; adverse effects ; Female ; Humans ; Middle Aged ; Myocardium ; metabolism ; Troponin I ; blood ; Young Adult

OBJECTIVETo investigate the effect of lipopolysacharide (LPS) in different concentrations on the biological features and growth factor secretion power of U937 cell line.

METHODSIn vitro cultured U937 cells were stimulated by 0 (as control), 0.1, 1.0, 10.0, 50.0 and 100 micro g/ml LPS respectively for 24 hours. Thereafter, the cell proliferation ability was determined by MTT method. The cell apoptosis rate was determined by flow cytometry. The changes in the contents of transforming growth factor beta(1) (TGFbeta(1)) and vascular endothelial growth factor (VEGF) of the supernatant of the cell culture were assessed by ELISA.

RESULTSApoptosis and TGFbeta(1) secretion could be induced by LPS in dose of 0.1 to 100 micro g/ml when compared with that without LPS challenge (P < 0.05 - 0.01). In detail, LPS in lower dose (0.1, 1.0 and 10.0 micro g/ml) could promote the proliferation of U937 (P < 0.05 - 0.01) but exerted no effect on VEGF secretion. In contrary, LPS in high dose (50 and 100 micro g/ml) could promote VEGF secretion (P < 0.01) but exerted no effects on the proliferation of U937 cells.

CONCLUSIONU937 cells could be activated to increase the secretion of TGFbeta(1) by LPS in optimal dose of 0.1 - 10.0 micro g/ml, but the secretion of VEGF could only be promoted by LPS in higher concentration.

Apoptosis ; drug effects ; Cell Division ; drug effects ; Humans ; Lipopolysaccharides ; pharmacology ; Transforming Growth Factor beta ; analysis ; secretion ; Transforming Growth Factor beta1 ; U937 Cells ; drug effects ; secretion ; Vascular Endothelial Growth Factor A ; analysis ; secretion