Background Establising the culture model of Müller cells for obtaining the highly putified target cells is essential for the study about the physiology and pathology of retinal Müller cells. The exsiting purifing method for culturing Müller cells is dissatisfactory. Objective This study was to establish a method to obtain high purifing Müller cells. Methods The retina from 5 clean newborn SD rats were isolated and digested by 0. 01% trypsin and cultured in DMEM containing 10% fetal bovine serum. The cellular suspension was then prepared,and the target cells were screened using flow cytometry based on the size and the quantity of cells. Cultured and passaged cells were identified by transmission electron microscope and light microscope. Immunocytochemistry was used to detecte the expression of GFAP in cultured cells for the determination of type and purity of the cells. Results The cells showed the similar shape to retinal Müller cells after primarily culture with the large volume, and some small other types of cells could been seen. The growth of cells was quickly 3 weeks later. The fibroblasts were removed using sticking-wall by steps,and neurons were eliminated following passage. Aboundent of cellular organs were seen under the transmission electron microscope. The positive response rate of the cells for CFAP was 100%. Conclution Flow cytometry offer a rapid and feasible approach for purifying Muller cell and it builds the foundation for further study about Müller cells.

Objective To explore the incidence ,clinical and neuroimaging features, possible risk factors and outcome of central nervous system (CNS) complications one year after hapilo-matched non-ablative hematopoietic stem cells transplantation. Methods The medical records of 36 consecutive patients who underwent hapilo-matched non-ablative hematopoietic stem cells transplantation for malignant and nonmalignant hematologic diseases between February 2004 and May 2009 were reviewed. Results CNS complications occurred in six (16.7 %) patients. Four of the six patients (66.7 %) died. Cerebral infarction occurred in three (8.3 %) patients. Cerebral softenness occurred in two (5.6 %) patients. Cerebral hemarrage occurred in 1 (2.8 %) patient. Epilepsy occurred in 1 (2.8 %) patient. The CNS complications occurred between 12 days and 286 days after stem cells transplantation. The age illness status and death rate were statistically different compared to patients without CNS complications (P ＜0.05). Conclusion The incidence and mortality of CNS complications are higher in those who underwent hapilo-matched non-ablative hematopoietic stem cells transplantation,which will make impacts on the patients' life quality and outcome.The illness status and eldly are probably the risk factors.

Rapid species identification of Mycobacterium tuberculosis by rpoB gene micro array.Based on the gene micro array of rpoB,the standard strains of 21 mycobacteria and 8 non-mycobacteria,126 clinical isolated of mycobacteria were detected by PCR-reverse dot blot hybridization assay.360bp DNA fragment was amplified from all mycobacteria tested and was not found in all nonmycobacteria except Hemolytic streptococcus and Corynebacterium pseudodiphtheriticum.The result of specimens were detected by the probe which is composed of 21 oligonucleotide was that probe-Mycobacterium fortuitum cross hybridizated with Mycobacterium platypoecilus while the other probes were specific.The 89 strains of the all 126 strains isolated from clinical specimens were identified to be mycobacterium tuberculosis,the percentage was 70.6%(89/126),while the other 9 strains were identified to be unmycobacterium tuberculosis and the the percentage was 9.2%(9/98).Identification of Mycobacteria by rpoB gene micro array is a rapid and effective method which is of considerable value in clinical territory.

OBJECTIVETo investigate the influence of down-regulating Smoothened (SMO) gene expression through short hairpin RNA (shRNA) on the proliferation of breast cancer stem cells.

METHODSHuman SMO shRNA was designed, synthesized chemically, and transfected into MCF-7 cells to down-regulate SMO gene. By using G418, stable cells with down-regulated SMO were selected. In vitro proliferation of these cells was measured by CCK8 assay. The proportion of CD44(+)/CD24(-) cells was detected by flow cytometry and the mammospheres formation was determined by suspension sphere culture. The expression of SMO, GLI1 and Oct4 was detected by Western blot. In vivo, the volume of tumor was measured every 3 days and the expression of SMO, GLI1 and Oct4 detected by Western blot.

RESULTSIn vitro, the cells were transfected with SMO-shRNA and selected by G418 after 21 days. SMO-shRNA effectively down-regulated the expression of SMO gene and protein, and inhibited the proliferation of MCF-7 and markedly reduced the proportion of CD44(+)/CD24(-) cells and mammospheres. In vivo, SMO-shRNA treatment of MCF-7 significantly inhibited the volume of tumor. The positive rate of SMO in negative control and SMO-shRNA group was 5/5 and 2/5, respectively. The expression of SMO, GLI1 and Oct4 in different groups were 0.72 ± 0.17 and 0.21 ± 0.09, 1.21 ± 0.21 and 0.47 ± 0.12, 0.83 ± 0.13 and 0.25 ± 0.07. SMO, GLI1 and Oct4 down-regulation significantly suppressed at protein levels (P < 0.05).

CONCLUSIONThe shRNA by chemical synthesis can effectively down-regulate SMO gene expression and inhibit the proliferation of breast cancer stem cells.

Animals ; Cell Proliferation ; Down-Regulation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Hyaluronan Receptors ; metabolism ; MCF-7 Cells ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplastic Stem Cells ; pathology ; Octamer Transcription Factor-3 ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Smoothened Receptor ; Transcription Factors ; metabolism ; Transfection ; Tumor Burden ; Zinc Finger Protein GLI1

OBJECTIVETo investigate the correlation of vasectomy with the risk of prostate cancer in Chinese men.

METHODSWe systematically searched the databases CNKI, VIP, Wanfang, PubMed, Embase, and Cochrane Library for the literature relating the relationship between vasectomy and the risk of prostate cancer in Chinese males up to December 2014. According to the inclusion and exclusion criteria, two investigators independently selected the eligible publications, evaluated their quality, and extracted relevant information, followed by a meta-analysis with the software STATA 12.0.

RESULTSNine studies were included in the analysis involving 1 202 cases of prostate cancer and 4,496 controls. Random-effect model analysis revealed no statistically significant correlation between vasectomy and the risk of prostate cancer (OR = 1.05; 95% CI 0.62-1.79), with an obvious heterogeneity (P < 0.001, I2 = 85.7%). No significant publication bias was found among the included studies (Egger, P = 0.824; Begg, P = 0.348).

CONCLUSIONThe results of our meta-analysis do not support the association of vasectomy with the increased risk of prostate cancer in Chinese population.

Asian Continental Ancestry Group ; China ; Humans ; Male ; Prostatic Neoplasms ; ethnology ; etiology ; Risk Assessment ; Vasectomy ; adverse effects

To review the development and application of powdering technique on oily drug of the traditional Chinese medicine. There have been numerous methods of powdering technique on oily drug, such as preparing complexation, microcapsule, adsorption by adsorbent, solid lipid nanoparticles, etc. And beta-Cyclodextrin complexation is the most usually operated. Powdering techniques have broad prospects in the pharmaceutical field, but more efforts should be made to improve oily drug of the traditional Chinese medicine.
Capsules ; Drugs, Chinese Herbal ; administration & dosage ; Nanotechnology ; Powders ; Technology, Pharmaceutical ; methods ; beta-Cyclodextrins

Objective To investigate the clinical diagnosis,treatment and prevention of fallopian tube prolapse(FTP)after hysterectomy.Methods A total of 7949 patients received hysterectomy from 1983 to Aug 2005 in Peking Union Medical College Hospital,including 6229 cases of trans-abdominal hysterectomy(TAH),780 cases of transvaginal hysterectomy(TVH),and 940 cases of laparoscopic assisted vaginal hysterectomy(LAVH).Nine cases(including 1 case from other hospital)of FTP after hysterectomy were analyzed retrospectively for their symptoms,diagnosis and treatment.All of them were diagnosed according to the results of histology and follow-up.Results The overall incidence of FTP after hysterectomy was 0.11%(9/7949).Incidence of FTP after trans-abdominal hysterectomy was 0.08% (5/6229),after vaginal hysterectomy 0.51%(4/780),and after laparoscopic assisted vaginal hysterectomy 0(0/940).There were no symptoms in 3 cases,but the other 6 cases had symptoms.The pelvic examination revealed the typical prolapsed fimbrial end of a fallopian tube in 3 cases and red granulation tissue in the other 6 cases.All of them were excised vaginally and cauterized.The results were confirmed by histological examination.No recurrent cases were reported in follow up.Conclusions FTP is a rare complication after hysterectomy.The prognosis is well after proper diagnosis and treatment.Salpingectomy or fixation of accessories into the pelvic wall and complete peritonealisation at the time of hysterectomy are important methods to prevent FTP after hysterectomy.

OBJECTIVETo establish the HPLC fingerprints of tanshinone microemulsion.

METHODHPLC analysis was carried on Hypersil C18(4.6 mm x 250 mm, 5 microm) analytical column; the acetonitrile and 0.5% phosphoric acid solution were used as mobile phases with gradient elution at a flow rate of 1.0 mL x min(-1). The UV detection wavelength was set at 270 nm.

RESULTSix peaks on HPLC fingerprint of Tanshinone Microemulsion are indexed.

CONCLUSIONThe method developed in the present study is convenient, reliable, and can be used as a quality control method for Tanshinone Microemulsion.

Chromatography, High Pressure Liquid ; methods ; Diterpenes, Abietane ; Emulsions ; Phenanthrenes ; administration & dosage ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Quality Control ; Salvia miltiorrhiza ; chemistry

iRGD-modified sterically stabilized liposomes loaded doxorubicin (iRGD-SSL-DOX) were prepared and their cellular toxicity and anti-tumor efficacy were evaluated, comparing to doxorubixin loaded sterically stabilized liposomes (SSL-DOX) and RGD modified doxorubixin loaded sterically stabilized liposomes (RGD-SSL-DOX). The iRGD peptide, with both tumor targeting and cell penetrating functions, was conjugated to DSPE-PEG-NHS and DSPE-PEG-iRGD was obtained. DSPE-PEG-RGD was gained in the same way. iRGD-SSL-DOX, RGD-SSL-DOX and SSL-DOX were prepared by ammonium sulfate gradient method. The size and zeta potential of the liposomes were characterized by dynamic laser light scattering. The cellular toxicity study was done on B16 melanoma cell line and the anti-tumor efficacy study was carried on B16 cell line bearing C57BL/6 mice. The results showed that the particle sizes of liposomes were all around 90-100 nm. DOX entrapment efficiency was above 95%. The formulations were with good preparation reproducibility. iRGD-SSL-DOX showed no significant difference in B16 cellular toxicity with SSL-DOX and RGD-SSL-DOX, but the anti-tumor efficacy on B16 melanoma bearing C57BL/6 mice was significantly better than that of SSL-DOX, similar as that of RGD-SSL-DOX. Therefore, iRGD modified liposomes loaded DOX would be a promising drug delivery system for tumor therapy.
Animals ; Antibiotics, Antineoplastic ; administration & dosage ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Doxorubicin ; administration & dosage ; pharmacology ; Drug Carriers ; Drug Delivery Systems ; Liposomes ; Male ; Melanoma, Experimental ; pathology ; Mice ; Mice, Inbred C57BL ; Molecular Weight ; Neoplasm Transplantation ; Oligopeptides ; chemistry ; pharmacology ; Particle Size ; Phosphatidylethanolamines ; chemistry ; Polyethylene Glycols ; chemistry ; Tumor Burden ; drug effects