The two-dimensional model of cell culture is an important method in the study of testicular development and spermatogenesis but can not effectively mimic and regulate the testicular microenvironment and the whole process of spermatogenesis due to the lack of relevant cell factors and the disruption of a three-dimensional spatial structure. In the past 20 years, the development and optimization of the in vitro model such as testis organotypic culture and in vivo model such as testis transplantation achieved a transformation from two- to three-dimension. The maintenance and optimization of the testicular niche structure could mimic the testicular microenvironment and cell types including Leydig, Sertoli and germ cells, which showed similar biological behaviors to those in vivo. Besides, the cell suspension or tissue fragment floats in the gas-liquid interface so that the development of somatic and germ cells is well maintained in vitro whilst the feedback linkage between grafted testis tissue and hypothalamus-pituitary of the host rebuilt in the in vitro model provides an endocrinological basis for spermatogenesis, which serves as an effective methodology to better understand the organogenesis and development of the testis as well as testicular function regulation, advancing the concept of treatment of male infertility. Al- though each of the methods may have its limitations, the progress in the processing, freezing, thawing, and transplantation of cells and tissues will surely promote their clinical application and present their value in translational medicine.
Cell Culture Techniques ; Germ Cells ; physiology ; Humans ; Infertility, Male ; therapy ; Male ; Spermatogenesis ; physiology ; Testis ; growth & development

Objective Study on the absorption,distribution,transformation,excretion and toxicity of four forms of arsenic in rats.Methods Four thousand and fifty data were obtained from 28 days animal metabolism experiments of 81 rats exposed to the four As species.Distribution,metabolism,excretion,and subacute toxicity of 4 As species were compared by analyzing the arsenate [As(V)],arsenite [As(Ⅲ)],methyl arsenate (MMAV) and dimethyl arsenate (DMAV) content of rat feces (excrement and urine),blood,seven organs,and liver and kidney pathology.Results After oral administration of each As species,82.9％ of As(Ⅲ),85.1％ of As(V),95.0％ of MMAV and 96.2％ of DMAV were accumulatively secreted via feces and urine,while 16.2％ of As(Ⅲ),14.1％ of As(V),4.65％ of MMAV and 0.120％ of DMAV were detected in blood.The DMAV accumulated in blood and urine after dosing As(Ⅲ),As(V) and MMAV,and the content of DMAV in the blood was 8 times greater than urine.Four kinds of arsenic had effects on rat liver and kidney function,the influence on liver was acute phase effect while that on renal was cumulative effects.The effect of As(Ⅲ) was the most significant,followed by MMAV.Four kinds of arsenic could cause tissue deformation and inflammatory cells infiltration,but showed no difference between groups.Conclusion The distribution,metabolism,excretion,and toxicity of the four kinds of arsenic in rats were different in varying degrees.The research of health risk for arsenic should consider its different forms.

Objective To evaluate the effect of various iterative reconstruction methods on phase analysis of gated myocardial perfusion imaging (MPI). Methods Thirty consecutive patients scanned by the Philips CardioMD system were recruited into this study. The gated SPECT (GSPECT) data were reconstructed with filtered backprojection (FBP),maximum likelihood expectation maximization (MLEM),three-di-mensional (3D) resolution recovery MLEM (AST),attenuation corrected (AC) MLEM,AC and 3D Monte Carlo scatter corrected (ACSC) MLEM methods. Parameters of left ventricular ( LV ) dyssynchrony ( phase standard deviation and histogram bandwidth) were measured using the software SyncTool. Paired t-test was used to compare the differences of the LV dyssynchrony indices between FBP and MLEM,AC MLEM,ACSC MLEM,AST respectively. Results The phase standard deviations of stress GSPECT MPI for FBP,MLEM,AC MLEM,ACSC MLEM,and AST were 11.6°,10.9°,11.2°,11.6°,11.4° respectively;while the histogram bandwidths were 35.7°,34.3°,35.1°,36.9°,35. 1 ° respectively. The phase standard deviations of rest GSPECT MPI for FBP,MLEM,AC MLEM,ACSC MLEM and AST were 15.2°,14. 5°,15.4° ,15. 4°,14.8° respectively; while the histogram bandwidths were 47.3°,46.4°,46.4° ,47.9°,46.1 ° respectively. No statistical significance was observed between the FBP and various iterative reconstruction methods for both the stress and rest GSPECT MPI study (t:-1. 179 to 1.554,P＞0.05 forall). Conclusion The standard FBP reconstruction method is accurate enough for the measurement of LV dyssynchrony indices using the widely used clinical software SyncTool.

AIM: To study caspase-9 expression on rat retina in the process of chronic elevation of IOP and the changes with the application of amino guanidine (AG), thus to investigate potential protective function of AG to rat retina with chronic elevation of IOP.METHODS: Immunohistochemistry, RT-PCR and Western blot were used to observe retinal morphology and expression of caspase-9 at different time points of rat with chronic IOP elevation, both affected or not affected by the application of AG.RESULTS: Compared with control group, as time passed retina of experimental group gradually had detectable morphological changes. On 21st day of chronic IOP elevation, retinas became thinner and the quantity of retinal ganglion cells (RGCs) decreased; caspase-9 expression increased, consistent with the morphological changes. The group using AG presented relatively smaller morphology changes and less expression of caspase-9.CONCLUSION: Apoptosis-related gene caspase-9 played a part in the process of chronic IOP elevation; AG protects retina by down-regulating expression of caspase-9.

OBJECTIVETo introduce a new procedure of laparoscopic adrenal surgery.

METHODSThirty patients with adrenal disease underwent surgery by a 70 degrees lateral decubital trans-abdominal laparoscopic approach, which wa compared with 30 degrees decubital laparoscopic operation in 26 patients.

RESULTSThe procedure is superior to open laparotomy. There was significant difference in average operative time (77.2 min vs. 215.7 min), blood lost volume (27.0 ml vs. 94.5 ml), and postoperative complications (13.3% vs. 46.0%). Postoperative feeding time and hospitalization time were shortened markedly.

CONCLUSIONSThis modified technique is safe, straightforward, with a good anatomic view during operation. It could be widely adopted for the treatment of benign adrenal disease, especially pheochromocytoma.

Adrenal Gland Diseases ; surgery ; Adrenalectomy ; methods ; Female ; Humans ; Intraoperative Complications ; prevention & control ; Laparoscopy ; methods ; Male ; Postoperative Complications ; prevention & control ; Posture ; Treatment Outcome

Objective To evaluate the accuracy of automated quantification of myocardial perfusion imaging (MPI) using a method based on a Western normal database for the detection of coronary artery disease (CAD) in a group of Chinese patients. Methods Seventy-two Chinese patients who underwent coronary angiography (CAG) and MPI within 3 months were recruited into this study. Eighty selected from 140 Chinese patients with low probability of CAD ( ＜ 5% ) were enrolled into local normal database of 99Tcm-methoxyisobutylisonitrile (MIBI) MPI using Cedars quantitative perfusion SPECT (QPS) database. Two Western MPI normal databases (CSMC MibiMbiAuto and Mibimibi) were used for processing the Chinese CAD patients recruited in this study, and the results were compared with those using local normal database and visual interpretation. T-test and z-test were used for statistical analysis. Results The extent (EXT)measurement obtained from Mibimibi and local database was ( 10.73 ± 14.54)% and ( 14.22 ± 16.51 )%,respectively ( t = 7.87, P ＜ 0.001 ); the severity (SEV) was 1.07 ± 0.93 and 1.34 ± 1.20, respectively ( t =7.45, P＜0.001). The area under curve(AUC) by using EXT measurement for local database (0.85 ±0.05) was larger than that for CSMC MibiMbiAuto ( AUC = 0.72 ± 0.06, z = 2.50, P ＜ 0.01 ) and Mibimibi ( AUC = 0.77 ± 0.06, z = 2.47, P = 0.014). The AUC of local database showed no significant difference from that of visual interpretation (AUC=0.83 ±0.05, z=0.05, P＞0.05). Conclusion Quantification of MPI of our Chinese patients using Western normal database would decrease the accuracy for the detection of CAD.

OBJECTIVETo study the effect on anti-respiratory syncytial virus of an active compound (AP3) from a Chinese medicinal herb-Herba patriniae in vitro.

METHODSActive component of herba patriniae (AP3) was extracted and its anti-respiratory syncytial virus (RSV) effect was tested. A water soluble substance (AP3) was isolated from a Chinese herb Herba patriniae, by hot water extraction, ethol precipitation and gel permeation column chromatography. The cytotoxicity of AP3 was tested by adding it to HeLa cells directly. Its effect against RSV was estimated by CPEI assay while ribavirin was used as positive control.

RESULTSChemical test showed that the nature of substance AP3 was polysaccharide. The median cytotoxic concentration (TC(50)) of AP3 was 11.45 mg/ml by morphological observation and the median effective concentration (50% effective concentration, EC(50)) of it against replication of the long strain of RSV in HeLa cells was 0.0986 mg/ml. The Therapeutic index (TI = TC(50)/EC(50)) of AP3 was 116.12, much higher than the TI of herba patriniae (AP1) (TI = 59.26) and ribavirin (TI = 53.45). Moreover, AP3 gave a dose-dependent response in inhibiting RSV. In the assay, the effect of AP3 against RSV growth was also tested. In addition, the effect of AP3 on virus growth, AP3 inhibited replication of RSV in HeLa cells, when added at 0 h, 2 h, 4 h after virus infection, were also tested.

CONCLUSIONThis study suggested that the AP3 exerted an obvious inhibitory effect to RSV in HeLa cell culture. This study furnished a reliable evidence for development of a new antiviral drug.

Antiviral Agents ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; HeLa Cells ; Humans ; Respiratory Syncytial Viruses ; drug effects ; Time Factors

To provide a better service for senior health care, we summarized screening studies of traditional Chinese anti-aging materia medica (TCAM). We collected and analyzed literature of TCAM screening studies using the lifespan test and animal models of aging from 1984 to 2012. We found 26 screening methods for TCAM, and 153 single herbs or active ingredients of TCAM that have been screened out during the past 28 years. The cell lifespan test, the fruit fly lifespan test, and D-galactose aging model were the most widely used and intensively studied screening methods. However, the method for establishing the D-galactose aging model needs to be standardized, and the D-galactose aging model cannot completely be a substitute for the normal aging mouse model. Great success has been achieved in screening studies in TCAM. To further improve screening studies in TCAM, we suggest that the D-galactose aging model be incorporated into the lifespan test in the New Drugs of Traditional Chinese Medicine Research Guide.
Aging ; drug effects ; Animals ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Materia Medica ; pharmacology ; Medicine, Chinese Traditional ; Models, Animal

The mechanism of apoptosis induced by SIRT1 deacetylase inhibitors in both human breast cancer MCF-7 and MCF-7 doxorubicin-resistant cells was studied. MTT assay was used to detect growth-inhibitory effect on the cells. Protein expression was detected by Western blotting. Chromatin condensation was detected by a fluorescent microscope after Hoechst 33342 staining. Cell cycle distribution was analyzed with flow cytometry. Apoptotic cells were detected with Annexin V staining. Nicotinamide (NAM) and Sirtinol, two SIRT1 deacetylase inhibitors, exhibited the similar growth-inhibitory effects on MCF-7/DOX cells and MCF-7 cells, but no potentiation of DOX activities. The arrest at G2/M phase was detected by flow cytometry in both MCF-7 and MCF-7/DOX cells after NAM treatment. Activation of caspase pathway in MCF-7 cells, such as the cleavages of PARP, caspase-6, -7, -9, were observed after exposure to NAM 50 mmol x L(-1), accompanied by the occurrence of chromatin condensation and Annexin V positive cells. However, the cleavages of PARP, caspase-6 and -7 in MCF-7/DOX cells delayed after exposure to NAM for 24 h and obviously increased at 48 h with appearance of chromatin condensation and Annexin V positive cells. SIRT1 deacetylase inhibitors show no cross resistance to MCF-7 drug-resistant cells, and the similar growth-inhibitory actions of them to MCF-7 sensitive and drug-resistant cells by which it is mediated by activation of apoptotic caspase pathway.
Apoptosis ; drug effects ; Benzamides ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Caspases ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Enzyme Inhibitors ; pharmacology ; Female ; Humans ; Naphthols ; pharmacology ; Niacinamide ; pharmacology ; Sirtuin 1 ; antagonists & inhibitors

AIMTo investigate the synergetic effect and the mechanism of antitumor action of the antibiotic lidamycin in combination with cisplatin in vitro.

METHODSCytotoxicity of the drugs was measured by clonogenic assay. Chromatin condensation was observed by co-staining with fluorescent dyes, Hoechst 33342 and propidium iodide. Apoptotic sub-G1 was detected by flow cytometry and DNA ladder was observed using agarose gel electrophoresis. Bcl-2 protein level was detected by Western blot assay.

RESULTSBy using clonogenic assay, lidamycin in combination with cisplatin was found to have synergetic effects on the proliferation of human hepatoma BEL-7402 cells. The data showed that BEL-7402 cells treated with cisplatin and lidamycin in combination produced internucleosomal DNA fragmentation analysed by agarose gel electrophoresis. The results of flow cytometry showed that cisplatin and lidamycin administrated in combination showed no obvious change in G1 phase distribution compared with single treatment. However, this combination reduced the S phase arrest and reversed the reduction of G2/M phase induced by single treatment. The results also showed that there was 11.3% or 9.37% of cells undergoing apoptosis in BEL-7402 cells treated with cisplatin or lidamycin, respectively, while it showed 32.4% of apoptotic cells in combination treatment. Cisplatin, lidamycin and combination of cisplatin and lidamycin was shown to induce typical chromatin condensation in BEL-7402 cells. The study showed that 0.5 mumol.L-1 cisplatin or 1 x 10(-4) mumol.L-1 lidamycin alone decreased Bcl-2 protein level, while lidamycin in combination with cisplatin strongly inhibited expression of Bcl-2 proteins in BEL-7402 cells.

CONCLUSIONThe results suggest that lidamycin enhancement of cisplatin-induced apoptosis associates with decrease of Bcl-2 protein expression, which may be useful for cancer chemotherapy.

Aminoglycosides ; pharmacology ; Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Cisplatin ; pharmacology ; Drug Synergism ; Enediynes ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; S Phase ; drug effects ; Tumor Cells, Cultured