Natural products are important sources for the discovery of anti-tumor drugs. Evodiamine is the main alkaloid component of the traditional Chinese herb Wu-Chu-Yu, and it has weak antitumor activity. In recent years, a number of highly active antitumor candidates have been discovered with a significant progress. This article reviews the research progress of evodiamine-based antitumor drug design strategies, in order to provide reference for the development of new drugs with natural products as leads.

Objective To study the genotypes of representative Mycobacterium tuberculosis (M.tuberculosis) strains from China with spacer oligonucleotide typing (spoligotyping),and to investigate the prevalence of different genotypes TB in China,and analyse the relationship between genotype and drug resistance.Methods 4017 clinical isolates were collected by Chinese Center for Disease Control and Prevention from 2007 to 2008 in 31 provinces in China according to sampling principle of epidemiology.Drug susceptibility testing was performed using proportion method,and spoligotyping was chosen to carry out genotyping of these M.tuberculosis.In addition,chi-square test was used to compare the differences among the detection rate of different genotypes.Results Among the 4017 M.tuberculosis isolates,2500 ( 62.2％ ) isolates belonged to Beijing genotype.The percentage of Beijing genotypes in the northern of China was higher than that in the southern of China ( 76.5％ vs.53.2％,x2 =219.69,P ＜ 0.05 ),while T1 genotypes were more common in the southern China,compared with that in northern China ( 13.3％ vs.4.3％,x2 =219.69,P ＜ 0.05 ).The differences were statistically significant.The proportions of Rifampinresistant (21.7％ vs.21.7％ ),Ofloxacin-resistant (4.9％ vs.2.4％ ) and Multidrug-resistant ( 11.3％vs.7.4％ ) isolates among Beijing genotype strains were significantly higher than those among non-Beijing strains (x2 =22.10,14.42 and 14.83,respectively,P ＜ 0.05 ).Conclusions Beijing genotype was still predominant epidemic genotypes.The percentage of Beijing genotype showed difference between distinct areas,and the percentage of Beijing genotypes in northern China was higher than that in southern China.Beijing genotype strains reveal correlation with Rifampin-resistance,Ofloxacin-resistance and Multidrug-resistance.

OBJECTIVETo investigate the application of free anterolateral thigh myocutaneous flap in the reconstruction of tongue and mouth floor defect after radical resection of tongue carcinoma.

METHODSFrom June 2006 to April 2009, 14 cases with tongue carcinoma underwent radical resection, leaving tongue and mouth floor defects which were reconstructed by anterolateral thigh myocutaneous flaps at the same stage. These 14 cases included tongue carcinoma at lingual margin (n=9), at ventral tongue (n=3) and at mouth floor (n=2). The flap size ranged from 7 cm x 9 cm to 5 cm x 7 cm.

RESULTSAll the 14 flaps survived completely with primary healing. There was no functional morbidity in the lower extremities. The patients were followed up for 12-26 months with satisfied esthetic and functional results in reconstructed tongue. Only one case (T4 N1 M0) died of metastasis carcinoma 14 months after operation. No local recurrence happened.

CONCLUSIONSThe anterolateral thigh myocutaneous flap has abundant tissue volume to reconstruct the tongue and mouth floor defect, while leaving less morbidity at donor site. Both satisfied esthetic and functional results can be achieved.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Mouth Floor ; surgery ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Thigh ; surgery ; Tongue ; surgery ; Tongue Neoplasms ; surgery

OBJECTIVETo establish a stable method of isolation, culture and cryopreservation of adult primary hepatocytes to provide potential hepatocyte resources for therapeutic usage in hepatocyte transplantation and bioartificial liver support systems for the treatment of acute and chronic liver diseases,and for experimental usage as an in vitro model of the liver.

METHODSAdult hepatocytes from 20 human donors undergoing partial hepatectomy were isolated using a two-step extracoporeal collagenase perfusion technique.Seven preincubation time points (2h,6h,12h,24h,36h,48h and 72h) were selected for optimization.After pre-incubation at 4 degrees C for 12-24h in HepatoZYME-SFM (the optimal condition),hepatocytes were microencapsulated using alginate-poly-L-lysine-alginate microcapsules,transferred to a complete medium containing 10% dimethyl sulphoxide and immediately placed into an isopropanol progressive freezing container for overnight freezing at -80 degrees C followed by immersion in liquid nitrogen the next day.During the post-thawing culture period,the cells were tested for albumin secretion,urea synthesis,cell cycling,transcription and protein synthesis (measuring mRNA and protein levels),and the morphological structure and pathology,for comparison with the features from before microencapsulated cryopreservation (PMC).

RESULTSThe viability and plating efficiency of the hepatocytes isolated using the two-step extracorporeal collagenase perfusion technique were 75.0+/-4.6% and 72.0+/-6.0%,respectively.The pre-incubation times of 12h and 24h (viability:61.4+/-4.8% and 62.0+/-5.6%; plating efficiency:3.2+/-5.8% and 62.6+/-3.6%,respectively) showed significantly higher albumin secretion than all other time points tested (F =40.3,all P less than 0.05).Compared with the immediate cryopreservation (immediately frozen control) hepatocytes,the PMC hepatocytes showed significantly better transcription and protein synthesis and higher albumin secretion and urea levels.The PMC group did not show a significantly different level of albumin production from the directly cultured hepatocytes (culture day 2:ll9.2ng/ml vs.131.36ng/ml,P =0.051; day 3:110ng/ml vs.120.4ng/ml,P=0.063; day 4:98.2ng/ml vs.109.8ng/ml,P more than 0.05).However,over culturing days 2,3 and 4,comparison of the PMC hepatocytes to the immediate cryopreservation hepatoeytes showed the former to have significantly higher secretion of albumin (119.2ng/ml vs.101.2ng/ml,110.0ng/ml vs.87.6ng/ml and 98.2ng/ml vs.73.8ng/ml; all P less than 0.05) and urea level (7.83 mug/ml vs.6.79 mug/ml,6.83 mug/ml vs.5.89 mug/ml and 5.85 mug/ml vs.4.83 mug/ml; all P less than 0.05).The post-thawed PMC hepatoeytes showed preservation of the morphological structure,while the immediate cryopreservation hepatocytes did not.

CONCLUSIONThe two-step extracorporeal collagenase perfusion technique after partial hepatectomy is a novel,simple,and reliable method for hepatocyte isolation.Pre-incubation at 4 degrees C for 12-24h before the microencapsulation cryopreservation allows for efficient recovery of functional and morphological integrity after thawing and provides viable hepatoeytes that may be useful for clinical applications in pharmacotoxicology,bioartificial liver therapy and cell therapy in humans.

Albumins ; Alginates ; Capsules ; Cell Cycle ; Cell Survival ; Cryopreservation ; Dimethyl Sulfoxide ; Hepatectomy ; Hepatocytes ; cytology ; Humans ; Perfusion ; Polylysine ; analogs & derivatives

Objective: To investigate the epidemical features, etiological and clinical characteristics of HFMD in Suzhou city, from 2011 to 2015, providing the scientific supports for HFMD prevention and control. Methods: In each district of Suzhou city, at least five specimens of mild cases were collected per month, while all of the severe cases were sampled. The RNA from each sample was examined using a commercially available real-time PCR kit. Statistical analysis was performed using SPSS version 11.5 software. Results: We retrospectively analyzed HFMD epidemiological data in Suzhou from 2011 to 2015, a total of 4 552 outpatients in Suzhou city were diagnosed with HFMD, including 2 818 positive specimen, the total positive rate was 61.90%, and there was a significant difference in the positive rates between the adjacent years (χ²=186.09, P<0.0001). From 2011 to 2015 in Suzhou, HFMD mainly affected children aged 1 to 5 years old, 66.17% of them were 1to 3 years old. Enterovirus 71(EV71) and coxsackievirus A16(CVA16) were the predominant viral genotypes in Suzhou from 2011 to 2012, and 2014. In 2013, other EVs were dominant, the severe cases mainly correlated with EV71 subtypes, and the proportion of patients with severe disease was significantly decreased in 2013. Conclusions EV71 and CVA16 are still the important pathogens of HFMD, other EVs also occupy a certain proportion, it is the time to select one or two advantage pathogens from the other EVs into the HFMD monitoring.

Objective To evaluate the efficacy and safety of motherwort (herbs leonuri/leonurus heterophyllus sweet) injection for preventing postpartum hemorrhage after caesarian section. Methods The prospective study was designed as a randomized and single blind multi-center research matched with positive agent as controls from Apt 2007 to Aug 2007. 440 women underwent caesarian section (CS) indicated by obstetric factors were enrolled from 15 teaching hospitals in China and assigned into three groups: group of motherwort: 147 cases were administered by motherwort 40 rag uterine injection during CS and 20 mg intramuscular injection per 12 hours 3 times after CS; group of motherwort + oxytocin : 144 cases were administered by motherwort 40 mg and oxytocin 10 U uterine injection during CS and motherwort 20 mg intramuscular injection per 12 hours 3 times after CS and group of oxytocin: 149 cases were administered by oxytocin 10 U uterine injection and oxytocin 10 U + 5% glucose 500 nd intravenously injection during operation and oxytocin 10 U intramuscular injection per 12 hours 3 times after CS. The following clinical parameter were collected and analyzed: (1) The amount of blood loss during operation, at 2, 6, 12, 24, 48 hours after operation. (2) The total amount of blood loss in 24 hours after CS and the incidence of postpartum hemorrhage. (3) The change of level of hemoglobin (Hb) and counting of red blood cell ( RBC ) from prepartum to postpartum. (4) Adverse reaction. Results (1) The mean amount of blood loss during operation were (368±258) ml in group of motherwort, (255±114) mi in group of motherwort + oxytocinand (269±141 ) ml in group of oxytocin, which exhibited significant difference among three groups ( P<0.01 ). Meanwhile, no statistical different amount of blood loss among three groups were observed at 2,6,12, 24, 48 hours after CS. (2)The amount of blood loss of postpartum at 24 hours were (480±276)ml ingroup of motherwort, (361±179) ml in group of motherwort + oxytocin, (381±179) nd in group of oxytocin, which showed significant difference among 3 groups(P <0.01 ). (3) The incidence of postpartum hemorrhage were 32.0% (47/147) in group of motherwort, 11.1% (16/144) in group of motherwort + oxytocin, and 18.8% in (28/149) in group of oxytocin. When comparing the lowest rate of postpartum blood loss in group of motherwort + oxytocin and the highest rate in group of motherwort, it displayed statistical difference (P<0.01). (4) The decreased level of RBC and Hb were shown that RBC(0.3±0.5)×10<'12<‘/L and Hb(9±13)g/L in group of motherwort, RBC (0.2±0.4)×10<'12/L and Hb ( 6±10) g/Lin group of motherwort + oxytocin and RBC (0.2±0.4)×10<'12/L and Hb(7±30) g/L in group of oxytocinrespectively. However, the comparison of different value of RBC and lib in group of oxytocin and motherwort +oxytocin showed significant difference (P<0.05 ). (5) Two cases with allery reaction was observed.Conclusion It is safe and efficacious that combined use of motherwort injection and oxytocin was to preventpostpartum hemorrhage during or after caesarian section.

The present study was designed to investigate the electrophysiological characteristics of rat conduit pulmonary artery smooth muscle cells (PASMCs) and the response to acute hypoxia. PASMCs of the 1st to 2nd order branches in the conduit pulmonary arteries were obtained by enzymatic isolation. The PASMCs were divided into acute hypoxia preconditioned group and normoxia group. Hypoxia solutions were achieved by bubbling with 5% CO2 plus 95% N2 for at least 30 min before cell perfusion. Potassium currents were compared between these two groups using whole-cell patch clamp technique. The total outward current of PASMCs was measured under normoxia condition when iBTX [specific blocking agent of large conductance Ca-activated K(+) (BK(Ca)) channel] and 4-AP [specific blocking agent of delayed rectifier K(+) (K(DR)) channel] were added consequently into bath solution. PASMCs were classified into three types according to their size, shape and electrophysiological characteristics. Type I cells are the smallest with spindle shape, smooth surface and discrete perinuclear bulge. Type II cells show the biggest size with banana-like appearance. Type III cells have the similar size with type I, and present intermediary shape between type I and type II. iBTX had little effect on the total outward current in type I cells, while 4-AP almost completely blocked it. Most of the total outward current in type II cells was inhibited by iBTX, and the remaining was sensitive to 4-AP. In type III cells, the total outward current was sensitive to both iBTX and 4-AP. Acute hypoxia reduced the current in all three types of cells: (1614.8+/-62.5) pA to (892.4+/-33.6) pA for type I cells (P<0.01); (438.3+/-42.8) pA to (277.5+/-44.7) pA for type II cells (P<0.01); (1 042.0+/-37.2) pA to (613.6+/-23.8) pA for type III (P<0.01), and raised the resting membrane potentials (E(m)) in all these three types of cells: (-41.6+/-1.6) mV to (-18.6+/-1.5) mV (P<0.01), (-42.3+/-3.8) mV to (-30.6+/-3.0) mV (P<0.01), (-43.3+/-1.6) mV to (-28.4+/-1.4) mV (P<0.01), for type I, II, III cells, respectively. These results suggest that acute hypoxia suppresses the potassium current and improves the E(m) in PASMCs. These effects may be involved in the modulation of constriction/relaxation of conduit artery under acute hypoxia. Different distribution of K(DR) and BK(Ca) channels in these three types of PASMCs might account for their different constriction/relaxation response to acute hypoxia.
4-Aminopyridine ; pharmacology ; Animals ; Calcium ; metabolism ; Cell Hypoxia ; Male ; Membrane Potentials ; drug effects ; Muscle, Smooth, Vascular ; cytology ; physiology ; Myocytes, Smooth Muscle ; physiology ; Peptides ; pharmacology ; Potassium Channels ; physiology ; Pulmonary Artery ; cytology ; physiology ; Rats ; Rats, Sprague-Dawley

BACKGROUNDA living fetus within the maternal uterus provides an example of allogene tolerance in mammals. Poria cocos Wolf is the main component of many Chinese medicinal combination drugs that have therapeutic effects on recurrent spontaneous abortion and that can maintain pregnancy until delivery. It was hypothesized that this herbal medicine can also prolong allograft survival after organ transplantation. Here, in an in vivo study, we report the anti-rejection effect of the ethanol extract of Poria cocos Wolf (EEPCW) in rats after cardiac allograft implantation.

METHODSTen normal rats were healthy controls. Eighty rats receiving homologous heart transplants were divided into 4 groups of 20 rats each based on type of treatment: olive oil 8 ml.kg(-1).d(-1), EEPCW 25 mg.kg(-1).d(-1), EEPCW 50 mg.kg(-1).d(-1) or cyclosporin A 5 mg.kg(-1).d(-1). Allograft survival was observed in 10 rats from each group. On the seventh day post transplantation, pathological lesions and percentages of CD3+, CD4+, and CD8+ lymphocytes and the CD4+/CD8+ ratio in peripheral blood were assessed in another 10 rats from each group and in 10 normal rats.

RESULTSThe survival time of donor hearts in the two EEPCW groups was significantly prolonged, to (15.9 +/- 2.4) days and (30.0 +/- 0.0) days, respectively, compared with (6.7 +/- 0.8) days in the control group. Pathological lesions in the two EEPCW groups were also less severe, and the percentages of CD3+, CD4+, and CD8+ lymphocytes and CD4+/CD8+ ratio were significantly lower in the EEPCW groups.

CONCLUSIONSAcute rejection of heart transplants and cellular immune reaction can be effectively suppressed using the EEPCW. Taking advantage of novel immunosuppressants derived from Chinese medicinal herbs used to treat abnormal pregnancy provides a hopeful road for future research and treatment in organ transplantation.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Graft Rejection ; prevention & control ; Graft Survival ; drug effects ; Heart Transplantation ; Immunosuppressive Agents ; pharmacology ; Male ; Polyporales ; chemistry ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar

OBJECTIVETo observe the protective effects on acute myocardial infarction of QDYX in dog.

METHODThe corconary ciculation and cardial oxygen metabolism, the degree and range of myocardial ischemia, myocardial infarct size, and the changes of the enzymes in serum were determined by using the acute myocardial infarction model of ligation of LAD in the anaesthetized open-chest dogs.

RESULTThe coronary resistance and cardial oxygen consumption were decreased and the myocardial blood flow was increased in dogs treated with QDYX of 1.0,2.0 mg.kg-1. The degree and range of myocardial ischemia, myocardial infarct size and the activity of serum CK, LDH were decreased in acute myocardial infarcion dogs treated with QDYX of 1.0,2.0 mg.kg-1.

CONCLUSIONQDYX can decrease cardial oxygen consumption in dogs, thus having protective effect on myocardial ischemia.

Administration, Oral ; Animals ; Astragalus membranaceus ; chemistry ; Cardiotonic Agents ; administration & dosage ; pharmacology ; Coronary Circulation ; drug effects ; Dogs ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Male ; Myocardial Infarction ; pathology ; physiopathology ; Ophiopogon ; chemistry ; Plants, Medicinal ; chemistry ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo construct anti-CD20scFv/CD80/CD28/zeta recombinant gene modified T cells, test its effectiveness of eradicating CD20+ lymphoma cells and provide a probably new approach to tumor adoptive immunotherapy.

METHODSCD28-zeta cDNA were amplified from vector pBULLET and inserted into pLNCX vector that contained anti-CD20scFv/CD80 gene. The recombinant vectors were transduced into PA317 cells and high titer retroviruses were obtained to infect human peripheral blood T lymphocytes. Resistant T cells were obtained by G418 selection at one week. Then transduced T lymphocytes and lymphoma cell lines Daudi Raji were cocultured. The cytotoxicity and cytokine production of transduced T cells were determined by non-radio-activation cytotoxicity assay and ELISA respectively.

RESULTSThe recombinant eukaryotic vector was constructed successfully as proved by enzyme digestion analysis and sequencing. These T cells were able to lyse CD20+ target cells and secrete high levels of IL-2 and IFN-gamma in vitro.

CONCLUSIONRecombinant gene modified T cells can be constructed successfully. It can specially kill CD20 positive lymphoma cells in vitro.

Antigens, CD20 ; genetics ; immunology ; B7-1 Antigen ; genetics ; immunology ; CD28 Antigens ; genetics ; immunology ; Cell Line ; Cytotoxicity, Immunologic ; Genetic Vectors ; Humans ; Immunotherapy, Adoptive ; Plasmids ; genetics ; T-Lymphocytes ; immunology ; metabolism ; Transfection