Objective To discuss the application of microsurgery in the treatment of male infertility.Methods From March 2007 to March 2012,there were totally 853 infertile men received microsurgical treatments in our department.Among them,344 patients with unilateral or bilateral varicocele underwent microsurgical varicocelectomy,60 underwent vasovasostomy (VV) and 192 underwent vastoepidystomy (VE)in microsurgical methods due to obstructive azoospermia.257 non-obstructive azoospermia (NOA) patients were performed microdissection of testicular sperm extraction (MD-TESE),at the same time,pathologic examination was done.Results ①For the varicocele patients,the pre-operative sperm density was (10 ±6) × 106/ml,the progressive sperm percentage was (16 ± 9)％.The post-operative density was (15 ± 8) ×106/ml,the progressive sperm percentage was (28 ± 14)％.The natural pregnant rate was 10.8％ (37/344).②In 60 patients undergone VV,the patent rate was 80.0％ (48/60),the natural pregnant rate was 35.0％ (21/60).In 192 VE patients,the patent rate was 53.1％ (102/192),the natural pregnant rate was 19.8％ (38/192).③In 257 NOA patients,the testicular volume,sperm retrieval rate of MD-TESE was significantly higher than that of conventional testicular sperm extraction (60.3％ vs.38.1％).Conclusion The microsurgery techniques in male infertility treatments could have some advantages such as explicit effects and decreased injuries.

Oroxylum indicum was a traditional Chinese medicine. In order to study the chemical constituents from the seed of O. indicum, the chemical constituents of 80% methanol extract of seeds of O. indicum were subjected to chromatography on silica gel, Sephadex LH-20, and preparative HPLC, leading to the isolation of eleven compounds. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR and 13C-NMR data as oroxin B (1), chrysin (2), baicalein (3), neglectein (4), quercetin-3-O-β-D-galactopy ranoside (5), quercetin-7-O-β-D-glucopyranoside (6), 2α,3β-dihydroxylluPeol (7), lupeol (8), rengyol (9), β-sitostero (10), and stigmasterol (11). Among them, compound 5 were firstly obtained from O. indicum.
Bignoniaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Seeds ; chemistry

OBJECTIVETo investigate reversal effect of histone deacetylase inhibitor LBH589 alone or in combination with proteasome inhibitor bortezomib on drug resistance in acute myeloid leukemia (AML) and its mechanism.

METHODSEx vivo cultures of HL-60/ADM cells and fresh refractory AML cells were treated with LBH589, bortezomib or their combination at varying concentrations. Proliferation capacity, apoptosis rate and reversal of drug resistance were evaluated by MTT assay, dual staining of Hoechst 33342 and Annexin VFITC/PI by flow cytometry, and adriamycin uptake rate with proliferation inhibition, respectively. The change of signal pathway at protein level was analyzed by Western blot.

RESULTSSynergistic cytotoxicity was observed in the combination treatment with LBH589 and bortezomib against HL-60/ADM cells, as well as the fresh AML cells, the most powerful synergy being observed at 21 nmol/L LBH589 plus 12 nmol/L bortezomib, with CI values of 0.531 and 0.498, respectively by Calcusyn software analysis. Moreover, the accumulation of adriamycin in HL-60/ADM cells was increased more in combination treatment [(64.81 +/- 3.69)%] than in either LBH589 [(28.96 +/- 2.52)%] or bortezomib [(37.29 +/- 3.71)%] alone (P < 0.05), and so did the uptake rate of adriamycin being (64.81 +/- 3.69)%, (28.96 +/- 2.52)% and (37.29 +/- 3.71)% respectively (P < 0.05). The combination treatment induced multiple apoptotic molecules co-action and intracellular drug accumulation contributed to the synergistic cytotoxicity, including caspase activation, PARP cleavage, XIAP downregulation, p53-dependent suppression of Bcl-2 and MRP1 expression via the inhibition of phosphoinositide 3-kinase (PI3K)/Akt/nuclear factor-kappaB (NF-kappaB) signaling pathway.

CONCLUSIONSCombination treatment of drug resistant AML cells with LBH589 and bortezomib produces a synergistic effect of in creating sensitivity to chemotherapy. The mechanism may be mainly resulted from inhibition of PI3K/ Akt/NF-kappaB signaling pathway.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; HL-60 Cells ; drug effects ; Humans ; Hydroxamic Acids ; pharmacology ; Indoles ; Leukemia, Myeloid ; genetics ; metabolism ; Pyrazines ; pharmacology ; Signal Transduction ; drug effects

INTRODUCTIONWe report a case of neurogenic pulmonary oedema (NPO) following massive left cerebral infarct, which was initially misdiagnosed as acute myocardial infarction (AMI).

CLINICAL PICTUREThis 52-year-old man presented with acute loss of consciousness with normal brain computed tomography (CT). He was treated as non-ST-elevation AMI complicated with pulmonary oedema based on findings of chest radiograph (bilateral pulmonary oedema), electrocardiogram (marked ST-T changes in leads V3 to V6), and cardiac enzymes [elevated creatinine kinase (CK) and CK-MB]. However, coronary angiogram and serial cardiac enzymes were inconclusive. Anisocoria developed after admission and a repeat brain CT was evident for large left cerebral infarct.

TREATMENTDecompressive craniectomy was carried out.

OUTCOMEMortality.

CONCLUSIONSThe diagnosis of NPO can be challenging when it occurs without abnormal findings on preliminary brain CT. It can be mistaken for cardiogenic pulmonary oedema secondary to AMI.

Cerebral Infarction ; physiopathology ; Coma ; Diagnosis, Differential ; Diagnostic Errors ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnosis ; Pulmonary Edema ; diagnosis ; physiopathology ; Radiography, Thoracic ; Taiwan

OBJECTIVETo investigate the effects of Fufang Banmao capusle on the proteome of SMMC-7721 cells and discover potential molecular mechanism of anti-cancer at molecular level.

METHODSMMC-7721 cells were treated by Fufang Banmao capusle serum prepared with serum pharmacological method; proteomic protocol involving 2-DE, image analysis and mass spectrometry were used to detect the proteins in cells influenced by Fufang Banmao capusle.

RESULTApproximately 450 protein spots in SMMC-7721 cells were resolved and detected in 2-D gel maps from pH3-10L IEF. 47 protein plots varied over 2-fold quantitively between treated sample and control sample were uncovered. 13 differentially expressed proteins spots were further identified by MALDI-TOF-MS analysis and four of them were successfully identified. Annexin A5, heatshock 70 x 10(3) protein 8 was significantly up-regulated in treated sample compared with control sample, while Eukaryotic translation initiation factor 5A and Peroxiredoxin-2 was significantly down-regulated in treated sample.

CONCLUSION4 differently expressed proteins associated with the proliferation, apoptosis, immunity of tumor were detected and they might provide clues for the coming research. The protocol of proteomics combined with serum pharmacological method is an effective platform to research complicated formulas in that it is capable of laying out many proteins associated with Fufang Banmao capusle.

Animals ; Annexin A5 ; metabolism ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Coleoptera ; chemistry ; Drug Combinations ; Electrophoresis, Gel, Two-Dimensional ; Female ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Materia Medica ; isolation & purification ; pharmacology ; Peptide Initiation Factors ; metabolism ; Peroxiredoxins ; metabolism ; Proteome ; drug effects ; metabolism ; Proteomics ; methods ; RNA-Binding Proteins ; metabolism ; Rabbits ; Serum ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Myelodysplastic syndromes (MDS) are heterogeneous clonal hematopoietic stem cell disorders with different mechanisms and diverse prognosis. The excess of ring sideroblasts (RS) is an important presentation MDS, but the mechanisms of RS appearance are obscure and the treatment of MDS-RS is intractable. Splicing factors play a very important role in the maturation process of eucaryon mRNA, recent studies indicate that there is a significant causal relationship between splicing factor 3B subunit 1 (SF3B1) mutation and the presence of ring sideroblasts. Lucubrating the downstream molecular of the mutated SF3B1 can facilitate exploring the mechanisms and new therapeutic strategies of MDS-RS.
Anemia, Sideroblastic ; etiology ; genetics ; Animals ; Humans ; Mutation ; Myelodysplastic Syndromes ; complications ; genetics ; Phosphoproteins ; genetics ; RNA Splicing Factors ; Ribonucleoprotein, U2 Small Nuclear ; genetics

The aim was to study the roles of extracellular regulated protein kinases (ERK) and telomerase activity in drug resistance of human leukemia and ovarian carcinoma cells. Flow cytometry was used to analyze apoptosis rate. Telomere repeat amplification protocol (TRAP) and bioluminescence analysis method were used for detection of telomerase activity. The phosphorylated ERK(1/2) protein expression was observed by Western blot method. The results showed that the specific inhibitor PD98059 of ERK kinase 1 (MEK(1)) enhanced the sensitivity of HL-60/E6 leukemia cell lines to harringtonine (HRT) or COC1/DDP ovarian carcinoma cell lines to cis-dichlorodiamine platinum (DDP). Both PD98059 and chemotherapy drugs HRT and DDP reduced the phosphorylated ERK(1) and ERK(2) protein expression level, and down-regulated the telomerase activity. The sole action of each was inferior to the combination action of PD98059 and HRT or DDP. In conclusion, ERK and telomerase serve a function to some extent in drug resistance of leukemia and ovarian carcinoma cells. The inhibition of ERK signal transduction pathways led to reduction of phosphorylated ERK(1) and ERK(2) protein expression level, and successionally down-regulated the telomerase activity. The final result was to enhance the sensitivity of HL-60/E6 to HRT or COC1/DDP to DDP.
Apoptosis ; drug effects ; Drug Resistance, Neoplasm ; Enzyme Inhibitors ; pharmacology ; Female ; Flavonoids ; pharmacology ; HL-60 Cells ; Humans ; Leukemia ; drug therapy ; pathology ; Mitogen-Activated Protein Kinases ; analysis ; antagonists & inhibitors ; Ovarian Neoplasms ; drug therapy ; pathology ; Telomerase ; metabolism

Objective To investigate the tolerated dose of the optic nerves and chiasm in patients with locally advanced nasopharyngeal carcinoma(NPC)treated with intensity-modulated radiotherapy (IMRT). Methods A retrospective analysis was performed on dose characteristics and the incidence of radiation optic neuropathy in 108 patients with locally advanced NPC treated with IMRT at D 2>55 Gy in the optic nerves and chiasm in our hospital between May 2009 and December 2013. The Common Terminology Criteria for Adverse Events(CTCAE)Version 3.0 grading criteria were used for evaluating adverse reactions of the optic nerves and optic chiasm.A logistic regression analysis was performed to assess the risk factors for the development of radiation-induced optic neuropathy(RION). Results No patient had severe RION (grade 3-5),although 7 of the 108 patients had mild optic nerve disorder(grade 1-2). No patient-or treatment-related factors were found to be associated with the development of RION(P>0.05). With a median follow-up of 46 months(range,13-91 months),the 3-year estimated overall survival,local recurrence-free survival,and distant metastasis-free survival rates were 90.0%,94.5%,and 86.4%, respectively. Conclusions The dose constraint of<55 Gy derived for optic nerves and chiasm from conventional radiotherapy does not seem to apply to IMRT. For advanced NPC patients treated with IMRT, the dose constraints of optic nerves and chiasm might be relaxed in order to improve target coverage.

Pathological cardiac hypertrophy induced by angiotensin II (AngII) can subsequently give rise to heart failure, a leading cause of mortality. Nardosinone is a pharmacologically active compound extracted from the roots of Nardostachys chinensis, a well-known traditional Chinese medicine. In order to investigate the effects of nardosinone on AngII-induced cardiac cell hypertrophy and the related mechanisms, the myoblast cell line H9c2, derived from embryonic rat heart, was treated with nardosinone (25, 50, 100, and 200 μmol/L) or AngII (1 μmol/L). Then cell surface area and mRNA expression of classical markers of hypertrophy were detected. The related protein levels in PI3K/Akt/mTOR and MEK/ERK signaling pathways were examined by Western blotting. It was found that pretreatment with nardosinone could significantly inhibit the enlargement of cell surface area induced by AngII. The mRNA expression of ANP, BNP and β-MHC was obviously elevated in AngII-treated H9c2 cells, which could be effectively blocked by nardosinone at the concentration of 100 μmol/L. Further study revealed that the protective effects of nardosinone might be mediated by repressing the phosphorylation of related proteins in PI3K/Akt and MEK/ERK signaling pathways. It was suggested that the inhibitory effect of nardosinone on Ang II-induced hypertrophy in H9c2 cells might be mediated by targeting PI3K/Akt and MEK/ERK signaling pathways.
Angiotensin II ; physiology ; Animals ; Cardiotonic Agents ; pharmacology ; Cell Line ; Cell Size ; drug effects ; Hypertrophy ; metabolism ; pathology ; MAP Kinase Signaling System ; Myoblasts, Cardiac ; cytology ; drug effects ; metabolism ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Sesquiterpenes ; pharmacology ; TOR Serine-Threonine Kinases ; genetics ; metabolism

To identify in vivo-induced gene in Streptococcus pneumoniae (S. pn) through a novel in vivo expression technology (IVET), a large promoter-trap library using galU and lacZ as the reporters was constructed. Based on the suicide vector pEVP3, a new vector pEVP3-galU was constructed with promoterless galU gene as an in vivo reporter. Firstly, promoterless galU gene was directly cloned into pEVP3 fusing with promoterless lacZ gene (an in vitro reporter). Then the random pieces of S. pn chromosomal DNA (200-500 bp), obtained by partial Sau3AI restriction digestion, were subcloned into the Bgl II site of pEVP3-galU. Upon introduction of the ligated plasmid library into E. coli DH5alpha by transformation, about 70,000 recombinants were recovered. Considering insert DNA orientation and insert size, this represents 5 coverage of the 2.2 Mb S. pn genome; 90% of these clones had 250- to 500-bp inserts. Thus, the library retained maximal complexity. Transformation by this plasmid library yield 450,000 S. pn transformants. The library was used to infect animals in intraperitoneal model. Those strains survived in vivo while exhibiting a white colony phenotype on TSA agar containing X-gal would indicate that the DNA fragment upstream of the galU reporter contained an in vivo-induced promoter. The promoter-trap library is suitable for screening in vivo-induced gene of S. pn.
Animals ; Cloning, Molecular ; Female ; Genes, Bacterial ; genetics ; Genes, Reporter ; genetics ; Genomic Library ; Mice ; Mice, Inbred BALB C ; Promoter Regions, Genetic ; genetics ; Streptococcus pneumoniae ; genetics