Objective To study the factor affecting the efficiency of establishment of parthenogenetic stem cells (PESCs)in mice. Methods PESC lines were derived from parthenogenetically activated blastocyst inner cell mass of different mice strains and cultured in different systems.Results The efficiency of establishing the pESC showed no significant difference in hybrid and inbred lines.But the efficiency was increased in culture systems added with ERK inhibitor or knockout serum replacement(KSR).Conclusion The efficiency of establishment of pESC was not directly related with the genetic background of mice but it was closely related to the culture systems.

Intrahepatic cholangiocarcinoma is the second most common intrahepatic primary liver tumor after hepatocellular carcinoma (HCC).Epidemiological study suggests a strong correlation between HBV infection and ICC development.This review focused on the potential mechanisms of HBV-induced ICC and gives a primary summary of suggested hypothesis,which included:(1) HBV infection of liver stem/progenitor cells will indirectly lead to HBV infection of intrahepatic biliary epithelial cells and lead to the development of ICC; (2) the changed microenvironment of intrahepatic biliary epithelial cells by HBV infection eventually results in carcinogenesis ; (3) the HBV infection of hepatic sten/progenitor cell can transform into tumor-like stem cells and ultimately differentiate into ICC-like tumor cells.

The subjects were all Chinese male adults. The average age, body weight and height were 37.9 (24-44) yr, 64.3 (51.5-74.0) kg and 173 (157-180) cm respectively in the first group, and 30.2 (24-41) yr, 67.7 (56.5-70.0) kg and 171 (170-176) cm respectively in the second group.The subjects in the first group were studied for 70 days for the N balance response at graded protein intakes of 0.42, 0.60, 0.73, 0.90 and 1.05 g/kg /day on Chinese mixed diet, during 5 experimental periods respectively. Each period took ten days preceded by one day of low protein diet, and was followed by three days of free choice diet The sequence of protein intakes during the study was 0.60, 0.73, 0.42, 0.90 and 1.05 g/kg/day. Six subjects in the second group were given a mixed diet ata protein level of 0.93 (0.91-0.94) g/kg/day for three months.In the N balance data of the first group, all subjects were in negative balance at protein intake of 0.42, 0.60 and 0.73 g/kg/day and all in positive N balance at 1.05 g/kg/day. Most of the subjects were in positive N balance at 0.90 g/kg/day. The linear regression analysis of N balance response of individual subject is that the intercept at zero balance of individuals ranged from 129.4-192.5 mg N/kg/day with the mean of the group of 147.7? 18.6mg N/kg/day or 0.92?0.12 g protein/kg/day. The five of the six subjects in the second group were in positive N balance, when on a mixed diet of 0.93 ?0.02 g protein/kg/day for three months.The results obtained indicated the mean protein requirement of these subjects based on the linear regression analysis of N balance response of individuals was 147.7 ? 18.6 mg N/kg/day or 0.92?0.12 g protein/kg/day. If 97.5% is to be covered, the safe level of protein intake should be 1.16 g protein/ kg/day.

BACKGROUND:Poultry mesenchymal stem cels are a particular subset of pluripotent adult stem cels derived from the mesoderm, which have great application prospects because of their strong proliferation and multi-directional differentiation potential. OBJECTIVE:To review the source, separation, purification, culture and differentiation of poultry mesenchymal stem cels, and to provide the theoretical foundation and experimental basis for the further research and application of poultry mesenchymal stem cels. METHODS:PubMed and CNKI databases were searched by the first author using key words of “mesenchymal stem cels, poultry, chicken, isolation, culture, differentiation” in English and in Chinese, respectively, to retrieve relevant articles published from 1990 to 2014. Literatures addressing induced poultry mesenchymal stem cels were included, and 42 articles were chosen for further analysis eventualy. RESULTS AND CONCLUSION: Poultry mesenchymal stem cels have great application prospects in the aspects of establishingin vitro model of poultry cels, studying poultry disease pathogenesis, animal nutrition and meat quality control. Its origin source is wide and easy to obtain. Isolation, purification, culture and biological characteristics of mesenchymal stem cels from different tissues are different. But, the study on poultry mesenchymal stem cels is stil in the exploration process, and there are many technical problems to be solved.

Intrahepatic cholangiocarcinoma (ICC) is a malignant liver tumor derived from the epithelial cells of the second- or higher-order branch of the bile duct, with the features of insidious early symptoms, high degree of malignancy, rapid disease progression, lymph node metastasis, intrahepatic metastasis, and vascular invasion, and such patients tend to have poor prognosis. Early diagnosis can improve the radical resection rate and prognosis of ICC. At present, the preoperative diagnosis of ICC mainly relies on imaging technology, laboratory examination, and pathological examination. In addition, some new diagnostic methods have also been used in the diagnosis of ICC in recent years, but histopathological examination remains the only method for the diagnosis of ICC. This article reviews the latest research advances in the diagnosis of ICC.

A reversed phase high performance liquid chromatography (HPLC) method was established for the simultaneous determination of 12,13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.A Grace Apollo C18 column (250 mm× 4.6 mm,5 μm) was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid (0.2％,v/v).Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃.An ultraviolet (UV) detector was used with a selected wavelength of 240 nm.Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12,13-dihydroxyeuparin (r＞0.9999) and 106.9-1068.9μg/mL for glycyrrhizic acid (r＞0.9999),respectively.Recoveries were 102.18 ％ for 12,13-dihydroxyeuparin and 101.17％ for glycyrrhizic acid.The method developed could be applied to the simultaneous determination of 12,13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.

ToprovidethemethodologyforrapidqualityevaluationofLonicerajaponica,wehaveestablished the stable quantitative model of near infrared spectroscopy ( NIR) . The performance of Bagging partial least squares (Bagging-PLS) model and Boosting partial least squares (Boosting-PLS) model was compared with that partial least squares ( PLS ) model based on the NIR data of ethanol precipitation process of Lonicera japonica. On this basis, the performance of these two models after variables selection was also studied by the methods of siPLS ( synergy interval partial least squares ) and CARS ( competitive adaptive reweighted sampling) . The experimental results showed that the prediction performance of Bagging-PLS and Boosting-PLS models was superior to PLS model with the latent factor of 10 . The band of 820-1029 . 5 nm and 1030-1239. 5 nm for the first batch was selected by the method of siPLS. In addition, the band of 820-1029. 5 nm and 1030-1239. 5 nm was selected for the second batch sample in the same method. Furthermore, the method of CARS was taken to select variables for the two batches samples with 5-fold cross-validation and 10-fold cross-validation. And the lowest RMSECV( root mean square error of cross-validation) values were used to take subset. Compared to the model performance without the method of CARS, the RMSEP value of the Bagging-PLS model and Boosting-PLS model for the concentration of chlorogenic acid reduced by 0 . 02-0 . 04 g/L and rp(correlation coefficient of prediction)value increased by 4%-5%. Generally, Bagging-PLS and Boosting-PLS could be regarded as rapid prediction methodsfor NIR quantitative models of ethanol precipitation process of Lonicera japonica.

Objective To evaluate the efficacy of intratympanic steroid therapy compared with systemic ster‐oid therapy on the treatment of idiopathic sudden sensorineural hearing loss (ISSNHL) patients with damaged glu‐cose tolerance .Methods Fifty first -diagnosed unilateral ISSNHL patients with damaged glucose tolerance were randomized devided equally to the intervention group (intratympanic steroid therapy) or the control group (systemic steroid therapy) ,all patients received conventional drug therapy simultaneously .Pure-tone hearing threshold tests were performed in all patients every 3 days after the first time ,and repeated measures anova was used to assess effects of hearing recovery accompanied with time .Results The mean hearing threshold in the control group de‐creased from 85 .4 ± 5 .6 dB to 48 .2 ± 4 .9 dB ,while in the intervention group it decreased from 84 .8 ± 5 .6 dB to 31 .7 ± 4 .6 dB .Total effective rate in the intervention group (84 .00% ,21/95) was higher than that in the control group (68 .00% ,17/25)(P<0 .05) .Conclusion The intratympanic steroid therapy is more effective than systemic steroid therapy in the treatment of ISSNHL patients with damaged glucose tolerance .

BACKGROUND:Atorvastatin has been shown to reduce bone loss and fracture, but its effects on implant osseointegration remain unknown.
OBJECTIVE:To investigate the effects of atorvastatin on implant osseointegration in osteoporotic rats and the underlying mechanisms.
METHODS:Forty-eight Sprague-Dawley rats were randomized into sham-surgery, ovariectomy, and atorvastatin (10 and 20 mg/kg per day) treatment groups, respectively. Al rats received ovariectomy and implant surgery except those in the sham-surgery group. Bone mineral density of the lumbar vertebra, osseointegration ratio and pul-out strength of implants were measured after 12-week treatment.Levels of bone formation and resorption markers in osteoblasts treated with atorvastatin were determined by ELISA. Wnt pathway-relatedgene expression was detected by RT-PCR.
RESULTS AND CONCLUSION:Bone mineral density, osseointegration ratio and pul-out strength of implants were significantly increased in 20 mg/kg per day of atorvastatin treatment group compared with ovariectomy group (P< 0.05). Levels of alkaline phosphatase, osteocalcinand osteoprotegerinwere significantly increased in osteoblasts treated with atorvastatinin vitro(P<0 .05), and the level of osteoclast differentiation factor RANKL was significantly inhibited (P< 0.05). Meanwhile, atorvastatin significantly promoted the mRNA expression of low-density lipoprotein associated protein 5and β-catenin, and inhibited the mRNA expression of dickkopfWnt signal pathway inhibitor 1and sclerostin. Our results suggest that atorvastatin promotes implant osseointegration in osteoporotic rats by activating Wnt/β-catenin signal pathway.

Objective To investigate the role of activities of sympathetic nerve in the pathogenesis of shoulder-hand syndrome (SHS) by analyzing the hand sympathetic skin response (SSR) at the acute stage of SHS after stroke. Methods 50 stroke patients with SHS at the acute stage were assigned as SHS group, another 50 stroke patients without SHS as control group (N-SHS group) and 50 health volunteers as health group. Every patient was subjected to the detection of bilateral hand SSR. Results The detection rates of SSR in the SHS group and N-SHS group were significantly lower than that in the Health group (P<0.01). In comparison within the SHS group, the amplitude of SSR on the affected hand was apparently higher than on the healthy hand (P < 0.05), but there was no significant difference in the SSR latencies in both hands ( P > 0 . 05 ) . In comparison with the health group , bilateral SSR latencies of the SHS group were longer than those of the health group (P<0.05) and bilateral SSR amplitudes were all lower than those of the health group (P<0.01). Conclusions The bilateral hand sympathetic skin responses could change at the acute stage of SHS after stroke, with possible presentations of central inhibition of sympathetic activity. The abnormality of SSR may be an early warning indicator in patients with hemiplegia after stroke complicated with SHS.