Research on the in vitro differentiation of embryonic stem cell(ES) is one of the hotspots in the study about developmental biology and cell transplantation therapy. But ethical issues and immune rejection remained to be the reasons which obstruct the progress. Recently, scientists have successfully obtained a kind of phripotent stem cell named induced phripotent stem cells (iPS). Since iPS posses similar properties with ES and can be autologously derived from patients themselves, it is a potential substitute of ES beth in the basic research and clinical application and the above mentioned problems can be avoided. However, iPS must differentiate to specific cells before being used in the clinical therapeutic research. The present article provides a concise summary of current progress on the differentiation ofiPS cell as well as some existing problems.

BACKGROUND: Reports of culture condition of olfactory ensheathing cells (OECs) vary. And some methods have bad reproducibility, not appropriate for actual application.OBJECTIVE: To observe the effect of differential attachment, chemicals in combination with trypsin digestion to purify rat OECs in vitro and to compare the effect with differential attachment or chemicals alone.DESIGN, TIME AND SETTING: In vitro controlled observation of cytology was performed at the Laboratory of Department of Human Anatomy & Histology and Embryology, Fujian Medical University from June 2007 to June 2008.MATERIALS: A total of 8 SD rats, 2 days old, were provided by the Laboratory Animal Center of Fujian Medical University.METHODS: The OECs were dissociated from the postnatal rat olfactory bulbs under sterile condition, and seeded in poly-L-lysine-coated culture flask at a density of 4.0×10~8 /L for primary culture. The cells were divided into 4 groups: control group (not purified); chemicals group (5 μmol/L arabinose); differential attachement group (Nash differential attachement); combination group (Nash differential attachement to eliminate most of the fibroblasts, followed by arabinose; when the cells were confluent at 6 days, the cells were digested with 1.25 g/L trypsin for 1 minute until the processes were shrank, cells became round, with some cells floating).MAIN OUTCOME MEASURES: Morphological changes of the cultured OECs and NGFRp75 immunocytochemistry were observed.RESULTS: The OECs displayed a very characteristic morphological appearance. Most of OECs were bipolar or tripolar with long and slim processes. In the unpurified control group the rapidly proliferating fibroblasts were in the majority (60%) after 7 days in culture, and confluent at day 14. The OECs occupied the most in the other groups, and their appearance remained unchanged.The surviving bipolar or tripolar OECs were positive for NGFRp75. The purifity by chemicals and differential attacehment methods was low (75%), while the combination group was high (85%).CONCLUSION: The method of purifing OECs through a combination of differential attacehment, chemicals and trypsinization is effective.

Objective To study the relationship among the slow vertex response (SVR) ,tone burst ABR threshold and pure tone thresholds in young people with normal hearing ,and to discuss the values of SVR and tb -ABR assessing hearing thresholds .Methods A total of 30 normal-hearing young people aged 20 to 24 were indud-ed in this study .The thresholds of SVR ,tb -ABR and threshold of PTA were tested at each verbal frequency (0 .5 ,1 ,2 ,4 kHz) .The differences and the relationship among SVR with PTA ,tb-ABR ,and PTA were statistical-ly studied .Results The differences between SVR and tb -ABR and PTA at various frequencies were statistically significant (P<0 .05) .The minimum difference between SVR and PTA was (3 .59 ± 7 .32) dB at 0 .5 kHz .The differences of 1 ,2 ,and 4 kHz were 6 .92 ± 5 .07 ,8 .33 ± 6 .71 ,and 9 .83 ± 9 .41 dB ,respectively .The minimum differences between tb -ABR and PTA was 14 .58 ± 7 .24 dB at 4 kHz ,and the differences of 0 .5 ,1 ,and 2 kHz were 29 .42 ± 6 .83 ,23 .25 ± 7 .47 ,and 16 .50 ± 6 .43 dB ,respectively .SVR and PTA had a good correlation at 1 and 2 kHz (r=0 .51 ,P<0 .01 and r=0 .44 ,P<0 .05 ,respectively) .tb-ABR and PTA had a good correlation at 4 kHz (r=0 .53 ,P<0 .01) .Conclusion In normal hearing young population ,SVR and tb-ABR can estimate PTA well , and the former is closer to PTA threshold .

Objective To investigate the value of single photon emission computed tomography (CT) imaging and transmis‐sion CT imaging (microSPECT‐CT ) bremsstrahlung imaging for the solid tumor mesenchymal implantaion of 32 P‐chromic phos‐phate‐paclitaxel‐poly‐L‐lactic acid (32 P‐CP‐PSP‐PLLA) sustained release seeds and to investigate the 32 P in vivo biodistribution and degradation sustained release churacteristics .Methods The animal model of prostate cancer subcutaneously transplanted tumor was established .32 P‐CP‐PSP‐PLLA sustained‐release seeds were intratumorally implanted by the mediation of microSPECT‐CT brems‐strahlung imaging and the 32 P distribution in bearing tumor mouse was verified by the imaging and biological distrubtion tests .The ultrastructural changes of 32 P seeds were observed by the scanning electron microscope .Results The MicroSPECT/CT brems‐strahlung imaging could effectively guide the intratumoral implantation operation of the 32 P sustained‐release seeds with clear visu‐alization .Partial sustained‐release 32 P was remained in the tumor tissues with little distribution in important organs of spleen and liver ,which was proved by the biodistribution results .The particle surface and inside micropores and tunnels formation ,their pro‐gressive increase ,fusion and connection were found by the electronic microscope after the 32 P sustained‐release seeds intratumoral implantation .Conclusion The MicroSPECT/CT bremsstrahlung imaging can effectively monitor the 32 P sustained‐release seeds and their in vivo biodistribution and lays a foundation for the sustained‐release seeds prostatic targeted implantation .

Objective To observe the suppression of special shRNA producing plasmid to hepatitis B virus(HBV) S gene and C gene on HBV replication and expression in HepG2.2.15 cells.Methods pSilenceCircle-U6 including pol Ⅲ promoter was used to construct HBV special shRNA producing plasmid as SC-S and SC-C.The experimental groups included SC-S group,SC-C group,unrelated control SC-N group and blank control group.With different dosages and at different time,shRNA producing plasmid was transfected into HepG2.2.15 cells.HBeAg and HBsAg in the culture media was detected by ELISA assay and HBV DNA in the culture media was measured by dot blotting assay. Results The recombinant shRNA producing plasmid with target sequence was constructed successfully.The inhibitory rates of HBeAg and HBsAg expressions by SC-S were much higher than those by SC-C.The inhibitory effects of HBeAg and HBsAg expressions were increased when the dose of SC-S was greater.The inhibitory effects of HBeAg and HBsAg expressions by SC-S were significant on the 3rd day after transfection and the inhibitory effect was the strongest on the 6th day.The inhibitory rate was still higher on the 9th day after transfection.Dot blotting assay showed the inhibitory effect of HBV replication by SC-S was greater than that by SC-C.Conclusion The shRNA producing plasmid with HBV S gene and C gene can be highly effective to inhibit the replication and expression of HBV.

BACKGROUND:The glass ionomer cement has good biological stability and can sustain the release of fluoride, which has been used as dental repair materials. But because of its low mechanical strength, the glass ionomer cement is confined to the front teeth and other parts with smal er force.
OBJECTIVE:To analyze the compressive strength, microleakage and other physical performances of Fuji IX glass ionomer cement after the addition of 8%modified nano-hydroxyapatite.
METHODS:(1) Universal testing machine was applied to detect the compressive strength of Fuji IX glass ionomer cement, Fuji IX glass ionomer cement added with 8%modified nano-hydroxyapatite, and super glass ionomer to fil the stainless steel cylinder. (2) Fuji IX glass ionomer cement, Fuji IX glass ionomer cement with 8%modified nano-hydroxyapatite and super glass ionomer were used to fil the tooth cavity;then, the microleakage was detected. (3) Fuji IX glass ionomer cement, Fuji IX glass ionomer cement added with 8%modified nano-hydroxyapatite, and super glass ionomer were used to fil stainless steel rectangular specimens. Then, universal material testing machine was applied to detect the bending strength value.
RESULTS AND CONCLUSION:The compressive strength, microleakage, and bending strength of specimens fil ed with Fuji IX glass ionomer cement the addition of 8%modified nano-hydroxyapatite were better than the other two groups, and Fuji IX glass ionomer cement was superior to the super glass ionomer in the compressive strength, microleakage and bending strength of fil ed specimens. It is shown that Fuji IX glass ionomer cement after addition of 8%improved nano-hydroxyapatite can improve the compressive strength and microleakage.

Objective To explore the efficacy and safety of the treatment of Xuezhikang and fluvastatin in elderly angina patients with mildly abnormal liver function .Methods 84 cases of ALT in 40-80u/L very elderly patients with coronary heart disease were randomly selected and divided into the two groups , the Xuezhikang group received Xuezhikang (0.6g,bid,orally),the fluvastatin group received fluvastatin (40mg,1time/night,oral),the total course was 12weeks,the TC,TG,LDL-C,HDL -C were measured before and after treatment ,liver function was measured once 2 -4times.If ALT was 3 times higher than before , Xuezhikang was instead of statins , if ALT was 5 times higher than before ,both of the two drags were withdrawaled .Results The TC,LDL,TG,HDL-C and ALT of the two groups were no significant difference before treatment (P >0.05),the TC,LDL,TG were decreased after treatment in the both group .There was no case of increased of ALT to three times in Xuezhikang group ,however ,ALT of 6 patients were increased more than three times in fluvastatin group ,four weeks after exchange of Xuezhikang ,the ALT were not continued to rise .Conclusion Both the two medicine can significantly lowered the cholesterol in very elderly patients with angina ,but the Xuezhikang were more safety in the patient with mild increased of liver function .

Objective ToobservetheefficacyofCO2lasercombinedwiththymopentinfortreatingcondylomaacuminata(CA)in anal canal .Methods 134 patients with CA in anal canal were selected and randomly divided into two groups .The treatment group was treated with CO2 laser for eliminating the wart combined with thymopentin by local injection ,while the control group was trea-ted with CO2 laser .Results The cure rate in the treatment group and the control group were 69 .1% (47 cases) and 45 .4% (30 ca-ses) respectively and the recurrence rate in the treatment group and the control group were 30 .9% (21 cases) and 54 .6% (36 cases) respectively ,the differences between the two groups had statistical significance (P<0 .05) .Conclusion CO2 laser combined with thymopentin has better effect for treating CA in anal canal .

BACKGROUND:There are many kinds of ways to obtain osteoblasts at present, but how to get high-purity osteoblasts in a easy and fast way has become a hot research.
OBJECTIVE:To explore a method to get massive and high purified osteoblasts effectively by comparing three common primary osteoblast culture methods, and to observe the biological characteristics of the osteoblasts from the skul of neonatal rabbit.
METHODCalvarias were dissected from newborn New Zealand white rabbits within 24 hours, and osteoblasts were isolated with bone tissue method, col agenase digestion method and modified tryptase and col agenase sequential digestion method respectively, then the cells were subcultured in vitro. Osteoblast proliferation and osteogenic activity were identified by inverted microscope for morphology observation. The rate of living osteobalsts was counted with trypan blue staining. The growth curve of the cells was drawn with MTT method. Alizarin red staining was applied to detect alkaline phosphatase and osteocalcin protein in the cellculture supernatants. Col agen I and col agen III immunohistochemical staining was also performed. RT-PCR was used to determine the expression of osteocalcin and col agen I mRNA expression.
RESULTS AND CONCLUSION:The cultured cells showed highly homogeneous appearance with active proliferation, and they had the typical features of osteoblasts. Alizarin red staining and col agen I immunohistochemical staining were both positive, while col agen III immunohistochemical staining was negative. Alkaline phosphatase and osteocalcin protein expression in the cellculture supernatants can be detected. The expression of osteocalcin and col agen I mRNA was positive in the RT-PCR test. Compared with col agenase digestion method, the modified tryptase and col agenase I sequential digestion method cost less time, presented higher production of osteoblasts and higher cellsurvival rate (P<0.05). Bone tissue method was the easiest method and did the least damage to osteoblasts, but it presented lowest production of osteoblasts and cost the maximum time among the three methods. So it cannot be used in large-scale osteoblast culture. A large quantity of high purity osteoblasts were obtained by modified trypsase and col agenase I sequential digestion method, which can be used as a reliable and efficient way to obtain the original generation osteoblasts in vitro.

Objective:To analyze the therapeutic effect and safety of finasteride on aged patients with isolated systolic hypertension (ISH)complicated benign prostatic hyperplasia (BPH).Methods:A total of 84 aged ISH + BPH pa-tients admitted from Jun 2011 to May 2013 were selected.According to medication plan,they were divided into fin-asteride group (n=44)and routine treatment group (n=40,only received routine treatment because they cannot use finasteride).Course of treatment was all six months.International prostate symptom score (IPSS),quality of life (QOL) questionnaire score,prostate-specific antigen (PSA),incidence rate of acute urinary retention,maximum u-rinary flow rate (Qmax),postvoid residual urine (PRV),prostate volume (PV)and incidence rate of adverse reac-tions were compared between two groups.Results:Compared with routine treatment group,there were significant reductions in IPSS [(13.2±2.4)scores vs.(9.4±1.1)scores],PRV [(21.4±4.6)ml vs.(9.0±2.3)ml],PV [(36.8±3.8)ml vs.(29.5±3.0)ml],PSA [(2.45±0.74)μg/L vs.(1.26±0.48)μg/L]and incidence rate of acute urinary retention (17.50% vs.9.09%),P<0.05 or <0.01,and significant increase in Qmax [(10.2±2.4) ml vs.(13.7±3.8)ml]and QOL score [(3.1±1.0)scores vs.(4.7±1.5)scores]in finasteride group (P<0.05). There were no significant difference in incidence rates of adverse reactions between two groups,P>0.05 all.Con-clusion:Therapeutic effect of finasteride is significant and adverse reactions don’ t increase significantly in aged pa-tients with isolated systolic hypertension complicated benign prostatic hyperplasia.