Aims To study the effects of clenbuterol on anoxia/reoxygenation( A/R) injury in neonatal Wistar rat cardiomyocytes and to explore whether its mecha-nism is related to reperfusion injury salvage kinase ( RISK) or not. Methods The cultured primary neo-natal cardiomyocytes were randomly divided into eight groups: ①normal culture group; ②anoxia/reoxygen-ation( A/R) group;③ clenbuterol ( 1 μmol · L-1 ) +A/R;④ICI118,551(10 μmol·L-1) + clenbuterol ( 1 μmol · L-1 ) + A/R; ⑤Metoprolol ( 10μmol · L-1 ) + clenbuterol(1μmol·L-1 ) + A/R group;⑥Metoprolol ( 10 μmol · L-1 ) + A/R group; ⑦PD98059 ( 20 μmol · L-1 ) + clenbuterol ( 1 μmol · L-1 ) + A/R group;⑧ LY294002(10 μmol·L-1 ) +clenbuterol(1 μmol · L-1 ) + A/R group. Cell via-bility was determined by the conventional MTT reduc-tion assay. The content of LDH in cultured medium was measured with colorimetry. Cardiomyocyte apopto-sis was determined by Hoechst33342 . Intracellular re-active species( ROS) were monitored by the fluorescent DCFH-DA. Total ERK2 and phosphorylated ERK were detected by western blot. Results Compared with A/R group, clenbuterol significantly increased vaibility of cells, reduced LDH release, lowered the rate of apop-tosis and ROS production. When addedβ2 receptor an-tagonist ICI118 , 551 , PI3 K inhibitor LY294002 and ERK inhibitor PD98059 , the effects of clenbuterol a-bove were inhibited; but β1 receptor antagonist Meto-prolol protected the cardiomyocytes from A/R injury, as evidenced by decreased LDH release and increased cell viability. There were no synergistic effects in the combined use of clenbuterol and Metoprolol. Conclu-sion clenbuterol exerts cardioprotective effects against A/R injury by inhibiting oxidative stress and apopto-sis. The protection of clenbuterol is inhibited by ICI118 , 551 , LY294002 and PD98059 . clenbuterol protects cardiomyocytes against A/R injury via RISK pathway by activation of β2 receptor.

Objective To study the efficacy of late course accelerated fractionation(LCAF) radio- therapy in the treatment of nasopharyngeal carcinoma(NPC).The end-po s were local control,radiation-in- duced complications,factors influencing survival.Methods From December 1995 to April 1998,178 NPC patients were admitted for radiation treatment.The radiation beam used was ~(60)Co?or 6 MV X-ray.For the first two-thirds of the treatment,two daily fractions of 1.2 Gy were given to the primary lesion ,with an interval of≥6 hours,5 days per week to a total dose of 48 Gy/40 fractions,over a period of 4 weeks.For the last one third of the treatment,i.e.beginning from the 5th week,an accelerated hyperfractionation schedule was carried out.The dose per fraction was increased to 1.5 Gy,2 fractions per day with an interval of≥6 hours,the total dose for this part of the protocol was 30 Gy/20 fractions over 2 weeks.Thus the total dose was 78 Gy in 60 fractions in 6 weeks.Results All patients completed the treatment.Acute mucosi- tis:none in 2 patients,Grade 1 in 43,Grade 2 in 78,Grade 3 in 52,and Grade 4 in 3 patients.Local control rate:the 5-year nasopharyngeal local control rate was 87.7%,and the cervical lymph node local control rate was 85.7%.The 5-year distant metastasis rate was 26.1%,and 5-year survivals was 67.9%. Sixteen patients had radiation-induced cranial nerve palsy.Conclusions With this treatment schedule, patient's tolerance is good,local control and 5 year survivals are better than control groups of conventional fractionation and hyperfractionation radiotherapy.Radiation-related late complication does not increase.Ran- domized clinical trials are being carried out to further confirm the efficacy of LCAF for nasopharyngeal carci- noma.

OBJECTIVETo research the constituents in needles of Pinus sylvestris.

METHODRepeated column chromatography and preparation HPLC are used for compound isolation, and their structures were elucidated on the basis of spectral data analysis.

RESULTSix compounds, pinifolic acid (1), 15-oxo-8 (17) -labden-18-oic acid (2) , 15-acetoxy-labd-8 ( 17)-en-18-oic acid (3), dehydroabietic acid (4), 7alpha-hydroxydehydroabietic acid (5), 7beta-hydroxydehydroabietic acid (6) were isolated from the needles of P. sylvestris.

CONCLUSIONCompound 3-6 were isolated from the needles of P. sylvestris for the first time, and compound 3 is a new natural product. The petroleum ether and EtOAc extracts showed significant cytotoxic effects to Hela and A549. Compounds 2, 4-6 revealed a positive distinction compared to the control group.

Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Chromatography, High Pressure Liquid ; Diterpenes ; chemistry ; isolation & purification ; pharmacology ; Diterpenes, Abietane ; chemistry ; isolation & purification ; pharmacology ; HeLa Cells ; Humans ; Molecular Structure ; Pinus sylvestris ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Structure-Activity Relationship

OBJECTIVEAn improved tracheal intubation method was used in preparation of myocardial infarction model in the mouse for increasing the success rate.

METHODSThe mice were intubated by an improved trachea cannula through oral cavity. The left anterior thoracotomy performed. The myocardial infarction model was made by legating the left anterior descending coronary artery in mouse. The color of heart was observed, and electrocardiogram was recorded. The survival rate and pathologic change were observed after two weeks of operation.

RESULTS40 myocardial infarction model mice were made by improved trachea cannula. The color of ventricles anterior wall had got madder red, and ST stages were raised on II leads of electrocardiogram in all of model mice. After two weeks, 27 mice were survival. The survival rate was 87.1% except for accidental death during operation. The heart chamber expanded and ventricular wall became thin in myocardial infarction mice by eyes. After pathological sections were stained, by HE cardiac muscle fibers ruptured or lysed. There were some of necrosis of myocardiac cells and many of infiltration of inflammatory cells.

CONCLUSIONApplication of an improved tracheal intubation method simplified operation of tracheal intubation during preparation of myocardial infarction model in the mouse. The trauma was tinier than the other one, and achievement ratio of the model preparation was improved.

Animals ; Disease Models, Animal ; Intubation, Intratracheal ; methods ; Male ; Mice ; Myocardial Infarction

OBJECTIVETo investigate and explain the composing mechanism by observing enhancement property of disassembled compositions of Yiqi Huoxue decoction (YQHXD) on platelet aggregation of health adults.

METHODVenous whole blood was obtained from healthy human subjects and anti-coagulated, and then centrifuged to produce platelet-rich plasma (PRP) and platelet-poor plasma (PPP). Disassembled compositions of YQHXD and sodium chloride were placed in different translucent tubes containing PRP and PPP, calculated the maximum percents of platelet aggregation stimulated by platelet agonists: adenosine diphosphate (ADP), platelet activating factor (PAF) and arachidonic acid (AA).

RESULTThe maximum percents of platelet aggregation induced by ADP and PAF were largely improved by YQHXD, both astragalus membranaceus and Angelica Scinesis played the most important roles in the decoction, the herbs of Yiqi and Huoxue must work together to enhance platelet aggregation.

CONCLUSIONIn vitro study, YQHXD can markedly improve platelet aggregation of health adults, but the exact mechanism should be proved by further experiments.

Adenosine Diphosphate ; pharmacology ; Adult ; Angelica sinensis ; chemistry ; Arachidonic Acid ; pharmacology ; Astragalus membranaceus ; chemistry ; Drug Combinations ; Drug Synergism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Humans ; Male ; Middle Aged ; Plants, Medicinal ; chemistry ; Platelet Activating Factor ; pharmacology ; Platelet Aggregation ; drug effects ; Young Adult

OBJECTIVETo investigate whether the alterations of microtubule and microfilament expression are responsible for the neurotoxicity of carbon disulfide.

METHODSWistar rats were administered with carbon disulfide by gavage at a dosage of 300 or 500 mg/kg for continuous 12 weeks (five times per week). Spinal cords of carbon disulfide-intoxicated rats and their age-matched controls were Triton-extracted and ultracentrifuged to yield a pellet and a corresponding supernatant fraction. Then, the contents of alpha-tubulin, beta-tubulin, and beta-actin in both fractions were determined by immunoblotting. In the meantime, their mRNA levels in spinal cords were quantified using reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSIn the supernatant fraction, the contents of beta-tubulin and beta-actin in both treated groups increased significantly (P < 0.01) the content of beta-tubulin increased by 141% and 158% respectively, and the content of beta-actin increased by 19% and 32% respectively. In the pellet fraction, the content of beta-tubulin in both groups increased by 107%(P < 0.01) and 118%(P < 0.01) respectively, and the others keep unaffected. In the meantime, the levels of of mRNA expression of beta-tubulin and beta-actin gene were elevated consistently in CS(2)-treated groups (P < 0.01) the levels of mRNA expression of beta-tubulin increased by 207% and 212% respectively, and the levels of mRNA expression of beta-actin increased by 94% and 91% respectively.

CONCLUSIONCarbon disulfide intoxication results in alternations of microtubule and microfilament expression, and the alternations might be related to its neurotoxicity.

Actins ; genetics ; metabolism ; Animals ; Carbon Disulfide ; poisoning ; Disease Models, Animal ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Spinal Cord ; drug effects ; metabolism ; Tubulin ; genetics ; metabolism

OBJECTIVETo investigate the effect of carbon disulfide (CS(2)) on oxidation-antioxidation function of rat nerve tissues.

METHODSThirty male Wistar rats were randomly divided into the control group, the low-dosage exposure group and the high-dosage group, 10 rats each. The rats of the two exposure groups were administered with CS(2) by gavage at a dosage of 300 or 500 mgxkg(-1)xd(-1), 5 times every week for continuous 12 weeks. The alterations in glutathione (GSH), malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), hydrogen peroxidase (CAT) and total anti-oxidation (T-AOC) in cerebrum, spinal cord, and sciatic nerve of CS(2)-treated animals were assayed.

RESULTSThe results showed that the contents of MDA and ROS in nerve tissues of CS(2)-treated groups increased significantly except ROS in spinal cord and sciatic nerve of low dose group. The content of MDA was increased by 20.7% and 33.6% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 18.5% and 23.3% respectively in the spinal cord, and by 20.7% and 53.0% respectively in the sciatic nerve, The content of MOS was increased by 20.1% and 34.9% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, and by 14.1% and 15.4% respectively in the spinal cord and the sciatic nerve of the rats of the high-dosage group (P < 0.05 or P < 0.01). Furthermore, the activities of SOD, GSH-Px, CAT and T-AOC decreased significantly except GSH-Px and SOD in cerebrum of low dose group. The content of GSH was decreased by 17.2% and 26.5% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 26.4% and 31.2% respectively in the spinal cord, and by 15.1% and 20.0% respectively in the sciatic nerve. The content of T-AOC was decreased by 11.1 and 26.4% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 15.1% and 38.4% respectively in the spinal cord, and by 35.6% and 42.3% respectively in the sciatic nerve. The activity of SOD was decreased by 12.1% and 25.4% respectively in the spinal cord of the rats of the low-dosage group and the high-dosage group and by 16.4% and 30.3% respectively in the sciatic nerve. The activity of GSH-Px was decreased by 17.3% and 32.5% respectively in the spinal cord of the rats of the low-dosage group and the high-dosage group and by 17.1% and 21.5% respectively in the sciatic nerve. The activity of GSH-Px and SOD was decreased by 12.6% and 30.1% respectively in the cerebrum of the rats of the high-dosage group. The activity of CAT was decreased by 17.5% and 39.4% respectively in the cerebrum of the rats of the low-dosage group and the high-dosage group, by 25.2% and 31.3% respectively in the spinal cord, and by 17.1% and 36.9% respectively in the sciatic nerve (P < 0.05 or P < 0.01).

CONCLUSIONSubchronic exposure to CS(2) can induce significant changes of oxidation-antioxidation function in rat nerve tissues, which might be related to CS(2)-induced neurotoxicity.

Animals ; Antioxidants ; metabolism ; Carbon Disulfide ; Lipid Peroxidation ; drug effects ; Nerve Tissue ; metabolism ; Rats ; Rats, Wistar

This study examined the expressions of miR-22 and miR-135a in rats with acute edematous pancreatitis (AEP) and their target genes in order to shed light on the involvement of miR-22 and miR-135a in the pathogenesis of acute pancreatitis (AP). The in vivo model of AEP was established by introperitoneal injection of L-arginine (150 mg/kg) in rats. The miRNA microarray analysis was used to detect the differential expression of miRNAs in pancreatic tissue in AEP and normal rats. The in vitro AEP model was established by inducing the rat pancreatic acinar cell line (AR42J) with 50 ng/mL recombinant rat TNF-α. Real-time quantitative RT-PCR was employed to detect the expression of miR-22 and miR-135a in AR42J cells. Lentiviruses carrying the miRNA mimic and anti-miRNA oligonucleotide (AMO) of miR-22 and miR-135a were transfected into the AR42J cells. The AR42J cells transfected with vehicle served as control. Western blotting was used to measure the expression of activated caspase3 and flow cytometry analysis to detect the apoptosis of AR42J cells. Targets of miR-22 and miR-135a were predicted by using TargetScan, miRanda, and TarBase. Luciferase reporter assay and quantitative real-time RT-PCR were performed to confirm whether ErbB3 and Ptk2 were the target gene of miR-22 and miR-135a, respectively. The results showed that the expression levels of miR-22 and miR-135a were obviously increased in AEP group compared with the control group in in-vivo and in-vitro models. The expression levels of miR-22 and miR-135a were elevated conspicuously and the expression levels of their target genes were reduced significantly in AR42J cells transfected with lentiviruses carrying the miRNA mimic. The apoptosis rate was much higher in the TNF-α-induced cells than in non-treated cells. The AR42J cells transfected with miRNA AMOs expressed lower level of miR-22 and miR-135a and had lower apoptosis rate, but the expression levels of ErbB3 and Ptk2 were increased obviously. It was concluded that the expression levels of miR-22 and miR-135a were elevated in AEP. Up-regulating the expression of miR-22 and miR-135a may promote the apoptosis of pancreatic acinar cells by repressing ErbB3 and Ptk2 expression in AEP.

Objective To compare the minimally invasive sinus tarsi approach and lateral intensive L-shaped approach in the therapeutic effects concerning medial wall reduction and calcaneal alignment for cal-caneal fractures. Methods A retrospective analysis was conducted of the 52 patients with calcaneal fracture who had been treated at Department of Foot & Ankle Surgery, Guangzhou Orthopaedic Hospital from January 2010 to December 2014. They were 39 men and 13 women, 28 to 46 years of age ( average, 40.4 years ). Of them, 26 were treated via the sinus tarsi approach ( minimally invasive group ) and the other 26 via the con-ventional lateral extensile L-shaped approach ( conventional group ) . X-ray axial films of the calcaneus were taken pre-operatively and post-operatively to evaluate the medial wall reduction and calcaneal alignment. The American Orthopedic Foot Ankle Society ( AOFAS ) ankle-hindfoot scale was adopted to assess the therapeutic effects. Results The average follow-up period for this cohort was 18 months ( from 12 to 24 months). The post-operative varus angle was 7.41°± 5.17°for the minimally invasive group and 8.01°± 5.33°for the con-ventional group; the correction of varus angle was 6.60°± 6.23°for the minimally invasive group and 8.57°± 6.64°for the conventional group; the good to excellent rate of medial wall reduction was 42.3% ( 11/26 ) for the minimally invasive group and 53.8% ( 14/26 ) for the conventional group; the AOFAS score was 89.5 ± 7.0 for the minimally invasive group and 86.2 ± 8.2 for the conventional group. There were no statistically signifi-cant differences between the 2 groups in all the above comparisons ( P ＞ 0.05 ). Conclusion The mini-mally invasive sinus tarsi approach can be a fine choice for treatment of calcaneal fractures, because it leads to no differences in medial wall reduction, postoperative varus angle and postoperative correction of varus angle, compared with the conventional lateral extensile L-shaped approach.

OBJECTIVESTo observe the change of erythropoietin (EPO) in patients of hypogonadism who received androgen replacement treatment and explore the mechanism of androgen-induced increase of red blood cells and haemoglobin.

METHODSEight patients with Klinefelter's syndrome, divided into two groups, received TU intramuscular injections of 500 mg or 1000 mg dose, respectively. After three months, seven patients received the second injection of crossover dose. Testosterone levels in serum were measured with RIA before and after the injections treatment. RBC count, impacted volume of blood cells and haemoglobin concentration were measured before treatment and 4, 8 weeks after treatment. At the same interval, EPO levels were measured with ELISA method.

RESULTSDevelopment of the secondary sex characters was improved in all patients after the TU injection. Serum testosterone levels raised significantly and reached the peak one week after the injections. Effective level of testosterone lasted for over 6 weeks. RBC count, impacted volume of blood cells and haemoglobin increased at different degrees after TU injections, but these changes were not significant in statistic(P < 0.05). The increased levels remained for 8 weeks. EPO levels were elevated significantly (P < 0.01 or 0.05) after the TU injection(Pbat > 0.05). The second injection could still make the EPO level go up.

CONCLUSIONSAndrogen replacement treatment can increase the EPO levels in patients of hypogonadism, which is one of the mechanism of RBC production increase.

Adolescent ; Adult ; Enzyme-Linked Immunosorbent Assay ; Erythropoietin ; blood ; Humans ; Injections, Intramuscular ; Klinefelter Syndrome ; blood ; drug therapy ; Male ; Radioimmunoassay ; Testosterone ; administration & dosage ; analogs & derivatives ; blood ; therapeutic use