Objective: To investigate the expression of ATP-Binding Cassette Proteins including P-gp (P-glycoprotein), MRP1 (multidrug resistance associated protein 1) and BCRP (breast cancer resistance protein) in hepatocellular carcinoma and its relationship with pathological features. Methods: The expression of P-gp/MDR1 (multidrug resistance gene 1), MRP1 and BCRP in hepatocellular carcinoma was examined by RT-PCR and immunohistochemistry in 34 cases of hepatocellular carcinoma and 19 cases of paraneoplastic hepatic tissues. Results: The expression of MDR1, MRP1 and BCRP mRNA (messenger ribonucleic acid) was 1.15±0.24, 0.64±0.33, and 1.07±0.32 in hepatocellular carcinoma and 0.36±0.14, 0.19±0.06, and 0.31±0.09 in paraneoplastic hepatic tissues. The expression of MDR1, MRP1 and BCRP mRNA was 1.38±0.26, 0.73±0.35, and 1.34±0.21 in poorly differentiated hepatocellular carcinoma and 0.74±0.32, 0.30±0.11, and 0.45±0.13 in well differentiated hepatic tissues. The immunohistochemical positive substance was detected in the plasma membrane and cytoplasm. The positive rates of P-gp, MRP1 and BCRP were 82.35%, 58.82%, and 79.41% in hepatocellular carcinoma and 42.11%, 26.32%, and 36.84% in paraneoplastic hepatic tissues, respectively. The positive rates of P-gp, MRP1 and BCRP were 100.00%, 81.25%, and 100.00% in poorly differentiated hepatocellular carcinoma and 66.67%, 38.89%, and 61.11% in well differentiated hepatic tissues. The expression of three indicies in hepatocellular carcinoma was higher than that in paraneoplastic hepatic tissues (P<0.05). The expression of P-gp/MDR1, MRP1 and BCRP in poorly differentiated hepatocellular carcinoma was higher than that in well differentiated hepatic tissues (P<0.05). No correlation was found among the three indices. Conclusion: Intrinsic multidrug resistance exsists in hepatocellular carcinoma, with various mechanisms. The multidrug resistance of HCC (hepatic cell carcinoma) is related to P-gp/MDR1, MRP1 and BCRP. MRP1 and BCRP may be targets for reversing multidrug resistance.

Objective To explore the possibility that rat bone mesenchymal cells (BMSCs) can differentiate into hepatocytes under the affection of fetal liver filtrate. Methods PAS and green indigo dye were used to detect glycogen and differential level of hepatocytes, respectively. The concentration of ALT, AST, ALP in the culture supernatant were served as markers of hepaocyte function. Results Fourteen days after induced by the fetal liver filtrate, BMSCs changed their shapes into polygon, oval or round. Some of BMSCs were positive for AFP and ALB at 7 days after induction, then the number of positive cells increased, and most of BMSCs expressed AFP and ALB till 21days. The PAS reaction and indocyanine green(ICG) intaking also appeared at 7days. Enzyme in supernatant such as ALT, AST, ALP were fristly detected at 7days and peaked at 14days,then the level declined. Conclusion The fetal rat liver filtrate was able to induce BMSCs into cells with function and characteristics of hepatocytes.

BACKGROUND: Studies of biological characteristics of mesenchymal stem cells (MSCs) and regulatory factors that influenced the differentiation of MSCs have shown that the proportion of the natural differentiation from in vitro primarily cultured MSCs into hepatocytes was low, and to select a suitable inductor is important to enhance the differentiation of MSCs into hepatocytes.OBJECTIVE: To verify the feasibility of induced differentiation of rat bone marrow MSCs (BMSCs) into hepatocytes using the combination of hepatocyte growth factor (HGF), epidermal growth factor (EGF) and fibroblast growth factor (FGF-4).DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Experimental Center, Weifang Medical College in August 2007.MATERIALS: Totally 40 Sprague-Dawley rats were supplied by the Experimental Animal Center, Weifang Medical College.METHODS: Rat BMSCs were incubated by adherent method. BMSCs at passage 3 were assigned to 2 groups. BMSCs in the blank control group were treated with L-DMEM containing 10% fetal bovine serum. BMSCs in the combination group were treated with 10 μg/L FGF, 8 μg/L HGF and 8 μg/L EGF following above-mentioned procedures.MAIN OUTCOME MEASURES: Inverted microscope was used to observe the morphological changes in cells.Immunofluorescence method was used to observe the expression of alpha-fetoprotein (AFP) and albumin (ALB). PAS was employed to detect the expression of glycogen. Fox green intake experiment was conducted. Enzymology was utilized to test the contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP).RESULTS: BMSCs in the combination group presented polygonal, orbicular or round shape. BMSCs in the blank control group remained spindle. BMSCs in the combination group were positive for AFP and ALB at day 14 following culture, and a few PAS-positive and fox green-positive cells were found at day 7. Positive cells became more over time. Synthesis of ALT, AST and ALP was detected at day 14, reached a peak at 21 days, and then decreased. Above-described indexes were negative in the blank control group.CONCLUSION: After induced by the FGF, HGF and EGF, BMSCs have the ability to differentiate into hepatocytes in vitro.

Objective To investigate the expression of hepatocyte nuclear factor-1α (HNF-1α) and hepatocyte nuclear factor-4α (HNF-4α) in human hepatocellular carcinoma (HCC) and explore the function of HNF-1α and HNF-4α during HCC carcinogenesis and development. Methods Twenty-six specimens of hepatocellular carcinoma were collected. The expressions of HNF-1α and HNF-4α in HCC tissues and adjacent non-cancerous tissues were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry staining. Results The mRNA levels of HNF-1α and HNF-4α were significantly lower in HCC tissues than that in adjacent non-cancerous tissues (0.818±0.371 vs. 0.383±0.102 for HNF-1α, P＜0.05;0.846±0.384 vs. 0.397±0.105 for HNF-4α, P＜0.05).The positive rates of HNF-1α and HNF-4α protein were significantly lower in HCC tissues than in adjacent non-cancerous tissues (92.3% vs. 42.3% for HNF-1α, P＜0.05;96.2% vs. 50.0% for HNF-4α, P＜0.05). The mRNA and protein expressions of HNF-1α and HNF-4α were correlated with tumor differentiation (P＜0.05). There was a negative correlation between HNF-1α and HNF-4α mRNA expressions in HCC tissues.Conclusion The expressions of HNF-1α and HNF-4α are down-regulated in HCC, which might be related to carcinogenesis and development of HCC.

Objective To investigate the value of individualized thyroid stimulating hormone (TSH) inhibition therapy in postoperative patients with differentiated thyroid carcinoma.Methods The medical record and follow-up data of the 556 patients with differentiated thyroid carcinoma after total or neartotal thyroidectomy were retrospectively reviewed.Patients were divided into two groups:Group A (304 cases) received TSH suppression therapy without risk assessment.Group B (252 cases) were given TSH suppression therapy in accord with risk assessment of both differentiated thyroid cancer recurrence risk stratification condition and the side effects of TSH suppression therapy risk stratification.Results The 3-year non-recurrence and (or) non-metastasis rate in group B was 99.2％ which was higher than 96.8％ in group A (P =0.044).The hospitalization rate caused by postoperative cardiovascular events or other morbidities in group B decreased 89％ than that in group A.Conclusions Individualized TSH suppression therapy can significantly decrease the recurrence and metastasis rate as well as concurrent morbidities caused by unnecessary TSH inhibition.

Objective To construct five shRNA-expression plasmids and to investigate the expression of Smad2 in TGF-?/ Smads signal transduction treated with shRNA-expression plasmid.Methods Five shRNA-Smad2 DNA sequences from mRNA sequence of mouse Smad2 gene were designed and synthesized.DNA oligonucleotides encoding an appropriate shRNA were inserted to shRNA expression vector respectively.Five shRNA-Smad2 expression plasmids were obtained and then transfected into NIH/3T3 cells.The suppressed expression of Smad2 was assessed by RT-PCR and Western-blotting.Results The shRNA-expression plasmid numbered 2.4 could markedly reduce the expression of Smad2.The suppression effect of the RNAi-pool composed of four different plasmids was more obvious than that of any single.Conclusion The shRNA-expression plasmids were successfully constructed,which could specifically and effectively suppress the expression of Smad2.The method of using a mixture of RNAi plasmids to improve the RNAi efficiency was established.

Objective To compare set?up error and the positioning and error correction time between the infrared markers automatic positioning+ ExacTrac ( A) and the manual positioning+ cone?beam computed tomography ( CBCT) image?guided radiotherapy ( IGRT) ( B) in intensity?modulated radiotherapy ( IMRT) for lung cancer. Methods A total of 20 patients with lung cancer were randomly divided into Group A and Group B. In Group A, after automatic positioning, a group of orthogonal X?rays images were taken using kV X?rays, which matched digitally reconstructed radiographs to obtain errors before correction. In group B, after manual positioning, images were taken using CBCT, which matched reference computed tomography images to obtain errors before correction. The positioning and error correction time was recorded in both groups. After error correction, errors after correction were obtained in each group using IGRT. Between?group comparison was made using the paired t test. Results The errors in lateral, longitudinal, vertical, and spinning vertical directions were significantly reduced after correction in both Group A and B (A:1.8±1?3 vs. 0.4±0?1, P=0?000;2.7±1?9 vs. 0.5±0?1, P=0?000;2.8±1?7 vs. 0.4±0?1, P=0?000;1.6±1?0 vs. 0.3±0?9, P=0?000;B:2.6±1?9 vs. 0.5±0?5, P=0?000;3.1±2?5 vs. 0.6±0?6, P=0?000;2.1±1?8 vs. 0.5±0?5, P=0?000;0.9±0?7 vs. 0.3±0?1, P=0?000). There were no significant differences in errors after correction between Group A and Group B (0.4±0?1 vs. 0.5±0?5, P=0?204;0.5±0?1 vs. 0.6± 0?6, P=0?257;0.4± 0?1 vs. 0.5± 0?5, P=0?518;0.3± 0?9 vs. 0.3± 0?1, P=0?755 ) . However, the positioning and error correction time in Group A was significantly shorter than that in Group B (199.1±16?2 vs. 315.2±13?7, P=0?000). Conclusions The application of ExacTrac or CBCT IGRT can substantially reduce set?up errors and improve set?up accuracy in IMRT. In addition, the application of the ExacTrac system can substantially shorten the positioning and error correction time.

OBJECTIVETo study the quantitative measurement of the extent of malrotation after interlocking intramedullary nailing of femoral shaft fracture.

METHODCT scan ("routine method") applied in 36 femoral shaft fractures that had been treated with close reduction and interlocking intramedullary nailing. For the judgement of the extent of malrotation, the anteversion of both fracture side and contralateral side were measured and the difference between the 2 sides was evaluated. The increase of anteversion represented internal rotation of the distal fragment, whereas the decrease of anteversion represented external rotation.

RESULTSThe maximum anteversion of the fracture sides, whereas 48 degrees, the minimum anteversion -10 degrees, the mean value, 15.04 degrees, and the standard error is 11.34 degrees. The maximum anteversion of the contralateral side, whereas 31.3 degrees, minimum -4.8 degrees, the mean value was 13.96 degrees and the standard error was 10.20 degrees (P < 0.001). Compared with the contralateral side, half of the 36 cases showed increased anteversion and the other half decreased anteversion. The mean value of internal rotation is 11.56 degrees, and external rotation 9.39 degrees. The maximum internal rotation was 37 degrees, the minimum 0.9 degrees. Eight cases had internal rotation less than 8 degrees, 6 between 10 degrees - 15 degrees, and 4 over 15 degrees. The maximum external rotation was 24.3 degrees, and the minimum 1.8 degrees. Eleven cases had external rotation less than 10 degrees, 4 between 10 degrees - 15 degrees and 3 over 15 degrees. The incidence of malrotation more than 10 degrees was 47% (17/36), and more than 15 degrees 19.4% (7/36).

CONCLUSIONThe incidence of malrotation after femoral shaft fracture treated with close reduction and interlocking intramedullary nailing is high. Attention should be paid to clinical management and strict control for rotational reduction intra-operatively.

Adolescent ; Adult ; Bone Nails ; Female ; Femoral Fractures ; diagnostic imaging ; pathology ; surgery ; Fracture Fixation, Intramedullary ; methods ; Humans ; Male ; Middle Aged ; Rotation ; Tomography, X-Ray Computed

OBJECTIVE To provide a reference for the safe use of drugs in patients with complex venous thromboembolism （VTE） and acute renal insufficiency. METHODS Clinical pharmacists participated in the management of anticoagulant therapy for a patient with complex VTE complicated with acute renal insufficiency， and evaluated the patient as high-risk thrombosis and bleeding based on their medical history， laboratory test results， etc.； combined with the complexity of thrombosis and renal insufficiency， clinical pharmacists suggested that enoxaparin sodium should be used in the acute stage of thrombosis （5 to 21 days after onset）， and then warfarin should be adopted for oral anticoagulation treatment. Because the patient’s anticoagulation was not up to the standard （the target range of the international normalized ratio was 2-3）， clinical pharmacists suggested increasing the warfarin dose， detecting the warfarin metabolism genotype， and adjusting the warfarin dose according to the genotype； at the same time， clinical pharmacists developed an anticoagulation monitoring plan to ensure the safety of anticoagulation treatment. RESULTS Doctors had adopted all the recommendations of clinical pharmacists. The patient did not experience adverse events such as bleeding or worsening of thromboembolism during anticoagulation in the hospital. When the anticoagulation met the standards， the patient was allowed to be discharged with medication. CONCLUSIONS By participating in the anticoagulation treatment management of patients with complex VTE and acute renal insufficiency， clinical pharmacists have assisted doctors in formulating personalized anticoagulation plans to promote the compliance with the anticoagulation treatment standard and ensure the safety and effectiveness of medication for patients.