Objective To isolate hepatic stem cells from adult rats,and to examine the process and morphologic changes of isolated hepatic stem cells differentiated into hepatic cells and bile duct epithelial cells under in vitro culturing conditions.Methods AAF/PH was used to stimulate the oval cells.Selective enzymatic digestion and density gradient centrifugation were employed to isolate and purify these cells from heterogeneous liver cell population.The hepatic stem cells were induced to proliferate and differentiate into hepatic parenchymal cells by the addition of dexamethasone,DMSO and several growth factors.The morphologic changes of hepatic oval cells to that of hepatic cells and bile duct epithelial cells were examined.Immunofluorescence method was used to detect the hepatic stem cells c-kit and the expression of OV6 antigen.The albumin of hepatic stem cells and the CK19 expression were identified by RT-PCR analysis.Results The activity of freshly isolated oval cells were over 90%,with different colors and size.The diameter of oval cells was about 1/4~1/6 that of hepatocytes.Large and round hepatocytes appeared first during differentiation,and the cell morphology changed when the oval cells differentiated to hepatocytes.Freshly isolated oval cells was positive for c-kit and OV6 with immunofluorescence staining,and RT-PCR analysis indicated that the albumin and CK19 mRNA were expressed in the new isolated cells.Conclusion The oval cells could be induced to differentiate into a variety of cell lineages including hepatocytes and bile duct epithelial cells in the presence of DMSO and several growth factors.Several antigens such as c-kit,OV6,CK19 and albumin may be expressed in the freshly isolated hepatic stem cells,and the course of morphologic changes may be observed under in vitro conditions of induction and differentiation.

The levels of cyclic nucleotides(CAMP and CGMP)in the serum and braest tissues of 60 patients with breast cancer or benign breast diseases were measured.It was found that the CAMP level was significantly lower,the CGMP significantly higher and the CAMP/ CGMP ratio was reduced in the serum and breast tissues of the patients with breast cancer.Our findings suggest that CAMP and CGMP might be factors of importance in the origin and development of breast cancer.

Objective To study the expression and cellular distribution of?-catenin in hepatocellular carcinoma and in HepG2 and SMCC-7721 cells.Methods Immunohistochemistry,immunocytochemistry techniques and Western blotting analysis were used to examine the expression and distribution of?-catenin in the cell membrane,cytoplasm and nucleus of bepatocellular carcinoma tissues,HepG2 and SMCC-7721 cells.Results Immunohistochemical staining showed that the?-catenin poorly,unevenly,even not distributed on the mem- brane of hepatocellular carcinoma cells.The expression of?-catenin on the membranes of low differentiated liver cancer cells was lower than that in the highly differentiated liver cancer cells,but it was strongly expressed in the cytoplasm of hepatocellular carcinoma cells,The ex- pression of?-catenin in the membranes of hepatocellular carcinoma cells was found to be reduced obviously in 18 out of 20 patients(90%).?-catenin was found to be poorly,unevenly,even not distributed in the membrane cells in 10 patients(50%).An obvious accumulation, but with poor-distribution,of?-catenin was observed in the cytoplasm of 17 cases(85%).Significant differences in the?-catenin expres- sion in both membrane and cytoplasm were found among differently differentiated stages of liver cancer cells.In 7 of 9 cases(77.8%)with highly differentiated liver cancer cells,?-catenin was continuously and evenly distributed in both the membrane and cytoplasm,while in 4 of 5(80%)cases with poorly differentiated liver cancer cells the expression of?-catenin in the membrane was found to be significantly poor and showed a uneven distribution in the cytoplasm.The?-catenin expression was positive in the cytoplasm of 5 of 6(83.3%)cases with moderatdy differentiated liver cancer cells.No expression of?-catenin was found in the membrane of HepG2 and SMCC-7721 cells,while it was highly expressed in both cytoplasm and nucleus.Western blotting showed that the?-catenin expressed in both cytoplasm and nucleus of all the liver cancer tissue,and also in HepG2 and SM(3C-7721 cells.Conclusions The expression of?-catenin was down-regulated in the membrane of hepatocellular carcinoma cells and HepG2 and SMCC-7721 cells,while?-catenin was strongly expressed in cytoplasm,and it was obviously accumulated in the nucleus.The distribution of?-catenin was related to the stage of differentiation of cancer cells.

Objective To study whether or not CD40 gene is a target of miR-503 in U937 cells,and to observe the regulatory effects of miR-503 on expression of CD40 induced by irradiation in U937cells.Methods The miR-503 sequence was inserted into pcDNA-DEST-47 plasmid to construct the eukaryotic expression vector (pcDNA-DEST-miR-503) and to construct the CD40 gene 3′-UTR luciferase reporter plasmid (psiCHECK2-CD40) at the same time.They were used to transfect U937 cells together for analysis of the regulatory effects of miR-503 on the expression of CD40.Meanwhile the miR-203 and miR-29b were used as controls to observe whether or not CD40 gene was a target of miR-503.The expression change of CD40 after irradiation,and the inhibitory effect of miR-503 on the expression of CD40 was confirmed by Western blot assay.The expression of miR-503 was detected by real-time RT-PCR (qPCR) after irradiation with doses of 5.0 Gy.Results The expression of luciferase in the group of transfected with pcDNA-DEST-miR-503 and psiCHECK2-CD40 plasmids was significantly lower than that in the group transfected with empty plasmid and CD40 gene (t =3.16,P ＜ 0.05).And the miR-203 and miR-29b could not inhibit the activity of CD40 luciferase,but the miR-503 could significantly inhibit the activity of it (t =5.25,P ＜ 0.01 ).The expression of CD40 protein in U937 cells was decreased after the cells were transfected with pcDNA-DEST-miR-503.After irradiation with dose of 5.0 Gy,the expression of miR-503 were increased (t =3.63 - 17.00,P ＜0.01 ) and the expression of CD40 protein decreased.Conclusions The CD40 gene might be the target of miR-503,and miR-503 could regulate the expression of CD40 gene.Irradiation could up-regulate the expression of miR-503 and inhibit the expression of CD40 protein in U937 cells.miR-503 might play a role in the process of regulation of irradiation on expression of CD40 protein in tumor cells.

Objective To evaluate the curative effect of different operative methods for duodenal diverticulum. Methods Articles of operations for duodenal diverticulum that were published between January 2000 to August 2006 were retrieved from China Journal Full-text Database, and the symptom remission rate and complication rate were underwent Meta-analysis. All data were analyzed by chi-square test. Results There was significant difference in symptom remission rate between Billroth Ⅱ subtotal gastrectomy, Roux-en-Y gastrojejunostomy and diverticulectomy, diverticulum inverting suture, sphincteroplasty of Oddi (χ~2= 21.269, 4.285, 12.914, 19.266, 3.938, P < 0.05). The difference in complication rate between Billroth Ⅱ subtotal gastrectomy, Roux-en-Y gastrojejunostomy and diverticulectomy, diverticulum inverting suture reached statistical significance (χ~2 =4.164, 9.166, 7.926,4.659, 4.858, P < 0.05). Conclusion For duodenal diverticulitis and the related complications which can not be effectively managed by internal medical treatment and with obvious symptom, Billroth Ⅱ subtotal gastrectomy and Roux-en-Y gastrojejunostumy are safe and effective.

BACKGROUND:Knee osteoarthritis is a chronic disease characterized as cartilage degeneration and hyperostosis to impact the soft tissues around the joints, thereby resulting in joint pain, and it can be aggravated and lead to joint deformity after weight-bearing. OBJECTIVE:On the basis of imaging parameters of knee osteoarthritis patients and normal controls, a model was established. And then, knee squatting movement was simulated on the model and stress data from the cartilage surface of patients and normal controls were colected and compared, thereby to obtain the stress characteristics of the cartilage surface in knee osteoarthritis patients. METHODS: There were 30 knee osteoarthritis patients and 30 healthy volunteers in the study. CT and MRI data from these participants were colected and analyzed using Mimics software and simpleware software to establish a model. Based on this model, cartilage-related stress data were harvested from knee osteoarthritis patients and healthy volunteers and compared to summarize the stress characteristics of the articular cartilage in patients with knee osteoarthritis. RESULTS AND CONCLUSION:In knee osteoarthritis patients, the stress on the cartilage surface from standing to squatting was increased in a nonlinear manner. During the process of squatting, the mediolateral stress was higher in the knee osteoarthritis patients than healthy controls (P < 0.05); however, there was no difference in the proximodistal and anteroposterior cartilage surface stress between the two groups (P > 0.05). These findings indicate that the knee osteoarthritis musculoskeletal model and finite element model are established successfuly, and these models are closer to the real motion characteristics of the knee joints, based on which, quantitative biomechanical data of the knee can be provided for knee osteoarthritis treatment.

Objective To investigate whether the sodium arsenite inhibiting proliferation of hepatocellar carcinoma(HCC) cells and having a relation with the expression of promyelocytic leukemia (PML) protein .Methods The immunohistochemistry , Western blot analysis ,immunofluorescence assay and quantitative PCR were used to examine the PML gene and protein expression in HCC tissue and cells .Results The immunohis to chemical staining showed that the PML protein was expressed in nucleus of well‐differentiated ,moderately differentiated and poorly differentiated HCC tissues randomly selected from 15 cases of HCC under‐going hepatic resection .Western blot analysis showed that PML protein was expressed at varying levels in all 24 HCC tissue sam‐ples ,HuH7 ,HepG2 ,Hep3B ,SMMC‐7721 and L02 cells .The immunofluorescence assay demonstrated that PML protein grains were found in the nucleis of the HuH7 ,HepG2 ,Hep3B and SMCC‐7721 cells ,in which HuH7 and Hep3B expressed more PML proteins than HepG2 and SMMC‐7721 cells .24 HCC tissue samples ,Hep3B ,HepG2 ,SMCC‐7721 and HuH7 cells all expressed PML gene .Sodium arsenite not only down‐regulated the PML protein expressed in HuH7 and primary HCC cells ,but also inhibited the HCC cell growth of HepG2 ,Hep3B ,HuH7 and SMMC‐7721 ,with the exposure time extension ,the inhibiting effect of sodium arsenite on HCC cells was enhanced .Conclusion Both HCC cell lines and tissues generally express ther PML protein and gene , PML protein may serve as the molecular basis for the direct targeting effects of arsenical agents .

Objective To observe the PML protein expression of hepatocellar carcinoma tissue and cells lines and As 2 O3 regulate its expression .Methods Immunohistochemistry was used to examine the PML protein expression of hepatocellar carcinoma tissue . Western blot analysis were used to observe PML protein expression of hepatocellar carcinoma tissue of 12 cases ,5 hepatocellar car-cinoma cell lines ,such as HuH7 ,HepG2 ,Hep3B ,SMMC-7721 ,MHC97H .Western blot analysis was used to detected the PML pro-tein expression of these hepatocellar carcinoma cell lines after 72-96 h treated with 0 .25 μg/mL of As2 O3 .Results Immunohisch-enmical staining showed that the PML protein was expressed in both cytoplasm and nucleus ,did not well-distributed in hepatocellar carcinoma cells .There was no significant differences of PML protein expressed among differently differentiated stages of hepatocel-lar carcinoma cells .Western blot analysis found that hepatocellar carcinoma tissues of 12 cases with hepatocellar carcinoma ex-pressed PML protein ,and there was significant difference of PML protein expressed among 12 cases suffer with hepatocellar carci-noma .hepatocellar carcinoma cell lines ,such as HuH7 ,HepG2 ,Hep3B ,SMMC-7721 and MHC97H all expressed PML protein ,and there was little difference of PML protein expressed among hepatocellar carcinoma cell lines .The PML protein expression of HuH7 ,HepG2 ,Hep3B ,SMMC-7721 and MHC97H cell after 72-96 h treated with 0 .25 μg/mL of As2O3 significant decreased . Conclusion Hepatocellar carcinoma tissue and cells may express PML protein ,and As2 O3 may regulate this protein expression as well .PML protein may be the target molecule of As2 O3 treating HCC .

BACKGROUND:The most effective method for the treatment of acetabular fracture is open reduction and internal fixation, however, this treatment for some special types of acetabular fracture cannot get satisfactory prognosis, and is prone to complications, such as traumatic coxarthrosis and avascular necrosis of femoral head. OBJECTIVE:To evaluate the curative effect of open reduction and internal fixation and total hip arthroplasty in the treatment of special acetabular fracture. METHODS:Twelve cases of acetabular fracture were included, including seven cases of traffic accident wound, three cases of crush injury, and two cases of fal ing injury. The type of bone fracture:two cases of posterior wal fracture, two cases of posterior column and posterior wal fracture, one case of T shaped fracture, five cases of transverse and posterior wal fracture, and two cases of acetabular roof sexual fracture. Complications:one case was femoral head centric dislocation, five cases were latter dislocation, and three cases were caput femoris fractures. Before injury, three cases had coxarthrosis, and two cases were avascular necrosis of femoral head. Al the cases were treated with open reduction internal fixation and total hip arthroplasty, of which nine cases were treated with biological prosthesis, and three cases were treated with bone cement prosthesis. The time from hospitalization to surgery was 3-15 days, and average was 6 days. The patients were fol owed-up once every 2 months in 1 year after replacement, and the Harris score was used to evaluate the hip function recovery. RESULTS AND CONCLUSION:No surgical site and deep wound infection, joint dislocation, lower limb deep vein thrombosis, and death were found in these 12 cases. Among them, 11 cases were fol owed-up for a longtime;the fol ow-up was lasted for 6-82 months. The acetabular fracture was healed at 6-16 months after reduction without prosthesis loosening and sinking. The hip function was evaluated during final fol ow-up according to the Harris score:excellent in eight cases, good in two cases, poor in one case, and the excellent and good rate was 91%. Open reduction internal fixation and one-stage total hip arthroplasty can avoid long-term bed, get out of bed as soon as possible, and reduce complications, thus reconstruct the hip joint painlessly and good functional y.

Objective: To explore the possibility of endoscope assisted curved laryngoscopy technique applied in transoral laryngopharyngeal minimally invasive surgery and evaluate the advantages of this technique. Methods: Eight patients with huge benign lesions in larynx and pharynx undergoing transoral microsurgery at Peking University Third Hospital between February 2016 and February 2017 were enrolled in this study.The diagnosis included cyst at the base of tongue in two patients, cysts in the epiglottis in two patients, hemangioma in two patients, multiple masses of the hypopharynx and amyloidosis in supraglottic area in one patient each.The time and exposure during surgery, occurrence rate of complication and conditions of following-up were recorded. Results: The mean time of surgery was 20 min, the exposure was satisfying.There was no obvious complication after surgery.No residual or recurrent lesion was observed after 1 to 12 months follow-up(mean time 4.5 months). Conclusion The endoscope assisted curved laryngoscopy technique has advantages in shortening the time of surgery, improving exposure and reducing the rate of complication and recurrence.