amount of bleeding to different degrees. It is believed that with the wide use of tranexamic acid during and after total knee arthroplasty, there wil be more optimal mode that can better control blood loss after total knee arthroplasty.

Diffusion tensor imaging (DTI)can reflect the brain structure and development more quantitatively and intuitively than conventional magnetic resonance sequence by obtaining the diffusion properties of water molecules. In neonatal brain developmental research,DTI could be used to study the developmental regularities of white matter tracts and cerebral structure deformity.It can also help to explore the relationship between white matter microstructure and neurodevelopmental outcome.In the study of brain injury,including premature white matter injury,hypoxic ischemic encephalopathy,neonatal stroke,and so on,DTI can diagnose the brain microstructure injury precisely,evaluate the ef-fectiveness of interventions and predict the long -term neurodevelopmental outcome.DTI may have good prospect in re-search and clinical application on neonatal brain development and injury.

Objective:To investigate L-type calcium channels of human mesenchymal stem cells(hMSCs) cultured in vitro.Methods:hMSCs were isolated,cultured by Percoll gradient centrifugation,adherence to plastic flask and monoclonal culture.Immunophenotypes of hMSCs were detected by immunofluorescence and flow cytometry techniques.Whole cell patch-clamp technique was used to observe the change of L-type calcium channels of human mesenchymal stem cells(hMSCs).Results:High homogenous hMSCs had been isolated and cultured in vitro.hMSCs had unique immunophenotypes and they were positive for CD44,CD29,c-kit but negative for CD34,CD31,CD54.Calcium currents could be found in 40 percent of hMSCs,peak currents of calcium(I Ca-Peak ) were (102.67?19.06)pA from +20 mV to +30 mV.The currents could be inhibited by Cd~ 2+ (20 ?mol/L).Conclusion:Undifferentiated hMSCs express less L-type calcium channes,regulation of the chanels might contribute to directional differentiation.

BACKGROUND: Reports regarding adipose-derived stern cells (ADSCs) differentiation into dopaminergic (DN) neurons are few in addition, there is not experimental evidence of the effect of ADSCs on maintaining the survival of DN neurons.OBJECTIVE: To investigate the effect of glial cell-derived neurokophic factor (GDNF)-modified adipose-derived stem cells on survival of DN neurons under co-cultured condition.DESIGN, TIME AND SETrlNG: The in vitro cytology experiment was conducted at the Institute of Otolaryngology-Head and Neck Surgery and Key Laboratory of Zoonoses of Ministry of Education between March and December 2007.MATERIALS: Wistar rats with 3-weeks-old, or 14 days of pregnancy were provided by Norman Bethune College of Medicine, Jilin University.METHODS: The GDNF recombinant adenovirus was constructed by using pAdTrackCMV and pAdEasy-1 system. DN neurons were obtained from the rostral mesencaphalic tegmentum of Wistar rat embryos by using trypsin and collagenase method. ADSCs isolated from rat inguinal fat pads were digested with collagenase Ⅱ, cultured and passaged in vitro. When the cells reached 60% cenfluency at the 3rd passage, cells were transfectad with 1×109vp/mL of Ad-GDNF for 1 hour and then transferred into growth medium for another 24 hours, and GDNF level in cell supematant was detected by ELISA assay. Meanwhile, the co-cultured of ADSCs and DN neurons were carried out for following 7 days. With GFP-modified ADSCs was served as a control group.MAIN OUTCOME MEASURES: The effect of co-cultured condition on the survival of DN neurons, as well as the differentiation of GDNF-modified ADSCs was detected by immunofluorescence staining.RESULTS: GDNF appeared in ADSCs supematant at 24 hours after Ad-GDNF transfection and reached a peak at 72 hours.There was approximately 80% GFP-positive labeled in ADSCs. The tyrosinase hydroxylase staining results demonstrated that the rate of survival DN neurons were significantly increased than in DA neurons cultured alone, co-cultured group of GFP-modified ADSCs and GDNF-modified ADSCs groups (55%, 15%, 25%, P < 0.01). However, there were no co-expressing TH and GFP positive cells appeared at 7 days of co-culture, which indicated that the co-cultured condition was not available to ADSCs differentiation.CONCLUSION: The co-cultured of GDNF modified ADSCs and DN neurons can promote the survival and growth of cultured DN neurons, however, it can not induce ADSCs differentiate into DN neurons.

Objective To isolate and cultivate hepatic stem cells(HSCs) from rat fetal liver in vitro and identify their biological features.Methods Collagenase perfusion method and mechanical cutting method were used to isolate HSCs from rat fetal liver which were then cultivated by H-DMEM containing 10% fetal bovine serum.The cell surface antigen expression of HSCs was observed with immunocytochemical method under confocal laser scanning microscope.Results The isolated HSCs from rat fetal liver grew to monolayer 5 d after cultivation in vitro.They presented pykno-round cells and distinct borderline under the light microscope.After 8 d the cells grew like epithelium.These cells expressed AFP antigen,CK 18 and CK19.Conclusion The cultivated cells are proved to be HSCs and can proliferate quickly in vitro.

Objectives To investigate the epidemiology of high-risk human papillomavirus (HPV) infection and the common genotypes in Liaocheng city, Shandong province, China, and to evaluate the application value of high risk HPV detection in cervical cytology with different pathological conditions.Methods A total of 19 707 permanent female residents in Liaocheng were recruited who were married or had sexual life, aged from 18 to 70 years old.They were screened for cervical cancer by thinprep liquid-based cytology test (TCT) from January 2013 to January 2014.The screen positive rate was 4.24 ％ (837/19 707), and 785 volunteers aged from 21 to 65 years old were recalled.The xMAP bead-based hybridization and flowcytometry analysis were used for genotyping.The data were analyzed by comparison and description.Results According to TCT, among 785 cases, there were 478 cases of atypical squamous epithelium of unknown significance (ASCUS) and atypical glandular epithelium of unknown significance (AGCUS), 175 cases of low-grade squamous intraepithelial lesion (LISL), 127 cases of high-grade squamous intraepithelial lesion (HSIL), 5 cases of squamous cell carcinoma (SCC) and adenocarcinoma (ACC).The positive rate of high-risk HPV was 62.8 ％ (493/785).The risk age of infection was 26-30 years old (87.7 ％, 71/81) and 51-55 years old (79.7 ％, 51/64), while a low risk one was found in patients older than 55 years old (28.6 ％, 14/54).The top five high-risk subtypes of HPV were HPV16 (21.5 ％, 169/785), HPV52 (12.2 ％, 96/785), HPV58 (9.8 ％, 77/785), HPV33 (9.7 ％, 76/785), HPV18 (7.5 ％, 59/785).Single infection accounted for 45.0 ％ (353/785), while multi-infection for 17.8 ％ (140/785).98 cases were infected by two subtypes, 37 cases by three subtypes, 2 cases by four subtypes, 2 cases by five subtypes and 1 case by six subtypes.Conclusions Compared with pure cervical TCT screening, high-risk HPV infection detection is an effective method for cervical cancer screening, which can improve the specificity of cervical cancer screening and reduce the omission diagnostic rate.In Liaocheng, HPV subtypes 16, 52, 58, 33, 18 and multi-infection are more prevalent.Women belonging to 26-30 or 51-55 years old are identified as high-risk population.Screening is important for this group to discover early cervical lesions.

OBJECTIVE To study the nephrotoxicity induced by bakuchiol alone and bakuchiol combined with psoralen and to explore its mechanism. METHODS The cytotoxicities of bakuchiol and bakuchiol combined with psoralen were investigated using human renal tubular epithelial cell lines (HK-2), in presence or absence of hepatic S9 mixture. The HK-2 cells were exposed to culture medium alone (blank control), 0.5% DMSO (vehicle control), aristolochic acid Ⅰ (AAⅠ;positive control), psoralen 5 μmol·L~(-1) group, bakuchiol 5,10,20,30 and 40 μmol·L~(-1) groups, and bakuchiol+psoralen (20+5), (30+5) and (40+5)μmol·L~(-1) groups, respectively. The cell viabilities were examined by MTT assay; cell membrane injuries were examined by detecting lactate dehydrogenase (LDH) release rate; and the morphological changes in HK-2 cells were observed with contrast microscope. The rate of cell apoptosis was detected by AnnexinⅤ/PI staining, and cell cycle was detected by PI staining with flow cytometry. RESULTS No cytotoxicity was found in psoralen 5 μmol·L~(-1) group. The HK-2 cell viabilities were significantly reduced after 4, 24, 48 and 72 h of exposure to either bakuchiol 20, 30 and 40 μmol·L~(-1)groups or bakuchiol+psoralen (20+5), (30+5) and (40+5)μmol·L~(-1) groups in a time- and concentration-dependent manner. The IC_(50) values of bakuchiol were (26.4±4.8), (21.8±0.6) and (24.1±0.8)μmol·L~(-1) for 24, 48 and 72 h exposure, respectively. The cytotoxicity of bakuchiol was significantly decreased in presence of hepatic S9 mixture. The LDH release rate of HK-2 cell increased significantly after 24 h of exposure to bakuchiol 20,30 and 40 μmol·L~(-1) or bakuchiol+psoralen groups. With the concentration and time increasing, the HK-2 cells became more and more contracted and rounded. In bakuchiol 40 μmol·L~(-1) or bakuchiol+psoralen (20+5), (30+5) and (40+5)μmol·L~(-1) groups, HK-2 cells showed apoptotic characters. In bakuchiol or bakuchiol+psoralen groups, apoptotic cells significantly increased and cells in G2 phase markedly decreased. CONCLUSION Bakuchiol has a significant cytotoxicity in HK-2 cells, and combined with psoralen can not decrease its toxicity. The cytotoxicity of bakuchiol is significantly reduced in the presence of hepatic S9 mixture. The possible mechanisms of the renal cytorotoxicity of bakuchiol are as follows: ① direct damage to the cell membrane; ② inducing cell apoptosis; ③ inhibiting intracellular DNA synthesis and block cell mitosis and proliferation.

The over-activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mechanistic target of rapamcyin (mTOR) pathway is closely related to the occurrence, development and clinical prognosis of malignant tumors. Taking this signal pathway as a target can effectively inhibit tumor progression. At present, the Food and Drug Administration of the United States has approved three drugs (CAL-101, BAY80-6946, IPI-145) for the treatment of recurrent and refractory indolent non-Hodgkin lymphoma, which demonstrates significant efficacy and a manageable safety profile.

Objective:To investigate the effect of interleukin-1β (IL-1β) on epithelial-mesenchymal transition (EMT) and cytoskeleton rearrangement of renal tubular epithelial cells.Methods:Immortalized renal tubular epithelial cell line NRK52E was cultured in vitro with IL-1β (30 μg/L) for 3 days and 6 days,then the cell morphology was observed;The mRNA expressions of α-smooth muscle actin (α-SMA),cytoskeleton components β-actin and α-tubulin were semi-quantitative examined by RT-PCR.The protein expression of α-SMA and arrangements of β-actin and α-tubulin were assessed by immunofluorescent staining.Results:After induced by IL-1β for 3 days and 6 days in vitro,the mRNA and protein expression of α-SMA increased significantly compared with corresponding control cells (P<0.001),it prompted that NRK52E cells underwent EMT;At the same time,the cell morphology also changed,from a typical multilateral paving stone to fibroblast-like appearance,with multiple processes; Cytoskeletal protein β-actin mRNA expression was also slightly increased (P<0.05).The distributions and arrangements of β-actin protein were also changed,from cell membrane transferred to peri-nucleus and cytoplasm,moreover it formed fiber bundle-like structures.However,another cytoskeleton protein α-tubulin in IL-1β induced cells,neither it's mRNA expression nor it's distribution had significant differences compared with the control group.Conclusion:IL-1β can induce NRK52E cells undergoing EMT in vitro,cell morphology changes into fibroblast-like appearance with multiple processes,and also the cytoskeleton protein β-actin expression increases and rearrangement occurrs.However,there was no changes onα-tubulin.

AIM: To investigate the expression of voltage-gated chloride channels (ClC)-3 protein and mRNA in human glioma specimen and its biological function. METHODS: The expression of C1C-3 was observed by immunohistochemical staining in 24 cases of human glioma, 4 cases of brain metastic cancer specimens and 5 cases of normal brain tissue as control; The C1C-3 mRNA expression were detected in the specimens with positive expression of ClC-3 protein by RT-PCR. RESULTS: ClC-3 protein was found negative in 4 cases of normal brain tissues and positive in 19 cases of human glioma and 4 cases of brain metastic cancer specimens. ClC-3 protein was mainly expressed in the membrane or cytoplasm of neoplastic cells and microvascular endothelial cells. The expression of ClC-3 mRNA was detected in 16 cases of human glioma and 4 cases of brain metastasis cancer specimens among the tissues with the positive expression of ClC-3 protein. The level of protein and RNA of ClC-3 in high malignant oligodendrogliomas was higher than that in low malignant ones. CONCLUSION: ClC-3 is generally expressed in human glioma and brain metastic cancer and is probably correlated with the classification of its pathological malignance.